PMID- 16466858 TI - Treatment of landfill leachate by Fenton's reagent in a continuous stirred tank reactor. AB - The treatment of landfill leachate by Fenton process was carried out in a continuous stirred tank reactor (CSTR). The effect of operating conditions such as reaction time, hydraulic retention time, pH, H(2)O(2) to Fe(II) molar ratio, Fenton's reagent dosage, initial COD strength, and temperature on the efficacy of Fenton process was investigated. It is demonstrated that Fenton's reagent can effectively degrade leachate organics. Fenton process reached the steady state after three times of hydraulic retention. The oxidation of organic materials in the leachate was pH dependent and the optimal pH was 2.5. The favorable H(2)O(2) to Fe(II) molar ratio was 3, and organic removal increased as dosage increased at the favorable H(2)O(2) to Fe(II) molar ratio. Temperature gave a positive effect on organic removal. PMID- 16466859 TI - Diffusion tensor imaging in acquired blind humans. AB - Retinal implants as a future possible therapy of blindness rely on an intact neural transmission from the retina to the primary visual cortex. By now it remains unknown, in how far the absence of afferent input in blindness affects also the organization of the optic radiation. Using diffusion tensor imaging (DTI), the non-invasive evaluation of large fiber tracts including the optic radiation has become possible. This method is sensitive to changes of the axonal state such as wallerian degeneration. We have compared DTI data from 6 acquired blind patients with those of a group of 11 healthy control subjects. Neither the relative anisotropy quotient of the visual fiber tract and the pyramidal tract showed a statistically significant difference between the blind patients and the control group nor did the absolute values of the relative anisotropy in the pyramidal tract and the visual fiber tract. There was no axonal degeneration of the optic radiation in late onset acquired blindness. With the optic pathways remaining intact, transmitting electric signals of retinal implants to the visual regions of the human brain seems to be possible even after decades of acquired blindness. PMID- 16466860 TI - Purification and characterization of an esterase isozyme involved in hydrolysis of organophosphorus compounds from an insecticide resistant pest, Helicoverpa armigera (Lepidoptera: Noctuidae). AB - An esterase isozyme was purified from the insecticide resistant pest, Helicoverpa armigera collected from field crops. Purification involved ammonium sulfate precipitation, hydrophobic interaction and ion exchange chromatography followed by gel filtration chromatography. The purification was 212-fold with 1% yield of the enzyme. The optimum pH of the isozyme was found to be 10.5 and 8.5 for p nitrophenyl phosphate and paraoxon, respectively. The enzyme was unstable at temperature >50 degrees C. The molecular mass determined by SDS-PAGE was 66 kDa. Cations such as Hg(+2), Ag(+2), Cd(+2) inhibited the activity while Zn(+2) stimulated it. Kinetic studies indicated that the enzyme had low K(m) values of 0.238 and 0.348 mM for p-nitrophenyl phosphate and paraoxon, respectively. The enzyme had broad substrate specificity with high K(m) values for ATP, ADP and beta-glycerophosphate. This enzyme was partially sequenced and identified as an alkaline phosphatase. PMID- 16466861 TI - Risk factors for alcohol misuse: examining heart rate reactivity to alcohol, alcohol sensitivity, and personality constructs. AB - OBJECTIVE: Heart rate reactivity to alcohol has been conceptualized as an index of alcohol-induced reward and has been associated with a sensation seeking personality profile. The goal of this study is to expand on previous findings regarding the significance of heart rate reactivity to alcohol while examining convergent lines of research on alcohol sensitivity, the rewarding effects of alcohol, and personality constructs. METHODS: Participants (N=47) were heavy drinkers who completed an intravenous alcohol challenge protocol. RESULTS: Analyses revealed a significant negative relationship between heart rate reactivity and alcohol-induced sedation and subjective intoxication. Heart rate reactivity was positively related to self-reported alcohol-induced vigor and to impulsivity and sensation seeking scores. CONCLUSIONS: Taken together, these results suggest that individuals with heightened heart rate reactivity to alcohol appear to be more sensitive to the invigorating properties of alcohol, while being less sensitive to the sedative and unpleasant effects of alcohol intoxication. These findings have implications to the conceptualization of heart rate reactivity to alcohol as a biobehavioral marker of alcohol sensitivity. PMID- 16466862 TI - Nicotine use and dependence and their association to psychiatric disorders in a large sample of adolescent psychiatric inpatients. AB - The purpose of this research was to evaluate the level of nicotine dependence (ND) and to examine its association to psychiatric disorders in a representative clinical sample of adolescent psychiatric inpatients. The modified Fagerstrom Tolerance Questionnaire (mFTQ) was used to assess the level of ND. Psychiatric DSM-IV diagnoses were obtained by using the Schedule for affective disorder and schizophrenia for school-age children (K-SADS-PL). Of the total of 342 inpatients in the study sample, 259 (75.7%) reported to be current smokers. A sum score 6 or higher in the mFTQ, indicating a high level of ND, was found in 37.9% of all smokers. An increased likelihood for high level of ND was associated with substance related disorders (OR 5.1, 95% CI 2.8-9.3), conduct disorder and oppositional defiant disorders (OR 2.4, 95% CI 1.4-4.4). The usefulness of mFTQ in measuring ND among adolescent inpatients is apparent. Therefore, it can be recommended to be used as a routine screening instrument for ND among adolescents hospitalized due to psychiatric disorders. PMID- 16466863 TI - Mental time travel in animals: a challenging question. AB - Humans have the ability to mentally recreate past events (using episodic memory) and imagine future events (by planning). The best evidence for such mental time travel is personal and thus subjective. For this reason, it is particularly difficult to study such behavior in animals. There is some indirect evidence, however, that animals have both episodic memory and the ability to plan for the future. When unexpectedly asked to do so, animals can report about their recent past experiences (episodic memory) and they also appear to be able to use the anticipation of a future event as the basis for a present action (planning). Thus, the ability to imagine past and future events may not be uniquely human. PMID- 16466864 TI - Experimental infection with Escherichia coli O149:F4ac in weaned piglets. AB - The outcome of experimental intestinal infections with enterotoxigenic Escherichia coli (ETEC) is dependent on several factors. An important factor is adhesion of the challenge strain to the intestinal mucosa. The test for susceptibility towards ETEC adhesion has so far been made by an intestinal adhesion test made after slaughter of piglets. However, in an experimental infection study with the purpose to obtain diarrhoeic piglets, it would be an advantage to test for susceptibility prior to experimentation. The Mucin 4 gene on porcine chromosome 13 has been proposed as a candidate gene for the production of the specific ETEC F4ab/ac receptor, and a DNA marker-based test has been developed to allow genotyping for ETEC F4ab/ac resistance/susceptibility [Jorgensen, C.B., Cirera, S., Archibald, A.L., Anderson, L., Fredholm, M., Edfors Lilja, I., 2004. Porcine polymorphisms and methods for detecting them. International application published under the patent cooperation treaty (PCT). PCT/DK2003/000807 or WO2004/048606-A2]. The aim of this study was to test an experimental model for ETEC O149:F4ac-induced diarrhoea in piglets, selected for susceptibility towards ETEC O149:F4ac adhesion prior to experimentation using a DNA marker-based test. Sixty-two healthy 25-32 days old recently weaned Danish crossbred piglets were used. All piglets were tested prior to experimentation for susceptibility or resistance towards ETEC O149:F4ac adhesion. Thirty-nine piglets, both susceptible and resistant, were oro-gastric intubated with 10(9)CFU of ETEC O149:F4ac and 23 age-matched piglets, both susceptible and resistant, were used as non-infected controls. Of susceptible piglets, challenged with ETEC O149:F4ac, 74% had ETEC O149:F4ac-associated diarrhoea first day after first challenge, which were significantly higher relatively to the resistant and challenged piglets where 20% had diarrhoea (p=0.04). This study suggests a model for experimental ETEC induced diarrhoea. PMID- 16466865 TI - Pushing the boundaries of viability: the economic impact of extreme preterm birth. AB - BACKGROUND: Previous assessments of the economic impact of preterm birth focussed on short term health service costs across the broad spectrum of prematurity. OBJECTIVE: To estimate the societal costs of extreme preterm birth during the sixth year after birth. METHODS: Unit costs were applied to estimates of health, social and broader resource use made by 241 children born at 20 through 25 completed weeks of gestation in the United Kingdom and Republic of Ireland and a comparison group of 160 children born at full term. Societal costs per child during the sixth year after birth were estimated and subjected to a rigorous sensitivity analysis. The effects of gestational age at birth on annual societal costs were analysed, first in a simple linear regression and then in a multiple linear regression. RESULTS: Mean societal costs over the 12 month period were 9541 pounds sterling (standard deviation 11,678 pounds sterling) for the extreme preterm group and 3883 pounds sterling (1098 pounds sterling) for the term group, generating a mean cost difference of 5658 pounds sterling (bootstrap 95% confidence interval: 4203 pounds sterling, 7256 pounds sterling) that was statistically significant (P<0.001). After adjustment for clinical and sociodemographic covariates, sex-specific extreme preterm birth was a strong predictor of high societal costs. CONCLUSION: The results of this study should facilitate the effective planning of services and may be used to inform the development of future economic evaluations of interventions aimed at preventing extreme preterm birth or alleviating its effects. PMID- 16466866 TI - Levels of 1-hydroxypyrene and other monohydroxy polycyclic aromatic hydrocarbons in children: a study based on U.S. reference range values. AB - Urine samples collected in 1999 and 2000 as part of the National Health and Nutrition Examination Survey (NHANES) were analyzed for 14 monohydroxy polycyclic aromatic hydrocarbons (PAH, metabolites of 7 PAH compounds) and for the first time reference range values were calculated for these metabolites in the U.S. population. The purpose of this paper is to explore differences in these PAH metabolites between children (6-11 years old), adolescents, and adults. More than 99% of the urine samples contained a detectable amount of 1-hydroxypyrene (1 OHpyrene), a metabolite of pyrene. We found that children in the youngest age group (6-11 years) had a geometric mean level (creatinine corrected data) 30% higher than children and adults in the other age groups, but no statistical differences existed between the two genders and among different racial groups. Smokers and persons exposed to environmental tobacco smoke (ETS) in 12-19-year old group and the 20-year-and-older group had higher levels of urinary 1-OHpyrene by a factor of 2-3 than non-smokers in the corresponding age group. Measurements of 3-hydroxyphenanthrene also suggested increased levels in children and in smokers. These results may indicate that young children are at a greater risk for PAH exposure, or that they absorb, distribute, metabolize, or eliminate PAH differently than adults. PMID- 16466867 TI - Sex steroid receptor expression in the oviduct and uterus of sheep with estrus synchronized with progestagen or prostaglandin analogues. AB - The objective of this study was to investigate differences in the expression of estrogen receptor-alpha (ERalpha), progesterone receptor (PR) and the proliferative indexes (Ki-67), in the uterus and oviduct of sheep with estrus synchronized either by prostaglandin analogues (Group PA, n=27) or by treatment with progestagens (Group P, n=29) on days 4 and 7 (day 0=estrus), when the embryos were collected. Immunohistochemical methods were used to quantify ERalpha, PR and Ki-67 in six superficial and deep compartments in the uterus and oviduct. The expression of ERalpha was significantly (P<0.01) lower in progestagen treated ewes than in prostaglandin analogues treated group in the luminal epithelium, superficial glands and superficial stroma in the uterus on day 4. The expression of PR was significantly lower in progesterone treated ewes than in the PA Group in the superficial gland (P<0.05) in both days studied. The lowest expression of PR was observed in the luminal caruncular epithelium and superficial glands in both treatments, obtaining the lowest levels on day 4 (P<0.05). There were significant differences between days 4 and 7 in the Ki-67 immunostaining in the luminal epithelium (P<0.01) and superficial glands (P<0.05). A higher cell proliferation was observed in the uterine epithelium (P<0.05) on day 4 in the animals treated with progestagens. Results indicate that sheep with synchronization of estrus with progestagens showed a reduction of ERalpha and PR protein expression in most of oviductal and uterine cells. PMID- 16466868 TI - Effect of altering dose of PG600 on reproductive performance responses in prepubertal gilts and weaned sows. AB - This study evaluated the effects of altering dose of PG600 on estrus and ovulation responses in prepubertal gilts and weaned sows. Experiment 1 tested the effects of one (1.0x, 400IU eCG+200IU hCG, n=74), one and a half (1.5x, n=82), or two (2.0x, n=71) doses of PG600 for prepubertal gilts. Estrus (58%) and ovulation (90%) were not affected (P>0.10) by dose. Higher doses increased (P<0.01) numbers of corpora lutea (17, 24, and 25), but not (P>0.10) the proportion of gilts with cysts (26, 36, and 46% for 1.0x, 1.5x, and 2.0x, respectively). Experiment 2 tested the effects of 0x (n=30), 0.5x (n=32), 1.0x (n=29), or 1.5x (n=30) doses of PG600 in weaned sows. Dose did not influence return to estrus (90%, P>0.10). There was an effect of dose (P<0.05) on incidence of cysts (3.4, 1.8, 6.4, and 29.8%, for 0x, 0.5x, 1.0x, and 1.5x doses, respectively). The 0.5x dose increased (P<0.01) farrowing rate (83.2%) compared to 0x (72.1%) and 1.5x (58.6%), but was not different from 1.0x (76.4%). Total pigs born (10.5+/-0.8) did not differ (P>0.10) among treatments. These data suggest that increasing dose of PG600 to 1.5x for gilts increases the number of corpora lutea but does not alter the proportion expressing estrus or ovulating. Reducing dose of PG600 for weaned sows did not alter estrus or ovulation, but the 0.5x dose increased farrowing rate compared to no PG600. PMID- 16466869 TI - Circadian variation in endothelial function is attenuated in postmenopausal women. AB - OBJECTIVE: The present study was undertaken to investigate the possible existence of an endogenously generated circadian rhythm in endothelial function in women and whether this rhythm is altered after the menopause. METHODS: Healthy non smoking women (11 pre-menopausal and 13 postmenopausal women) were studied during a 22 h period under constant routine conditions; endothelium-dependent (flow mediated dilation (%FMD)) and -independent (glyceryl-trintrate (GTN)-mediated) function was assessed every 2 and 4 h, respectively, by high-resolution ultrasound of the brachial artery. RESULTS: %FMD and %GTN was significantly higher in pre-menopausal women (9.9+/-1.0%FMD (mean+/-S.E.M.); 18.2+/-1.8%GTN; P<0.01) compared with postmenopausal women (6.5+/-0.5%FMD; 11.5+/-1.6%GTN). A significant day-night variation in %FMD was observed pre-menopausal women (day 9.2+/-0.8%; night 10.4+/-1%; P<0.05) with an attenuated rhythm in postmenopausal women (day 6.8+/-0.6%; night 6.0+/-0.4%). CONCLUSIONS: The findings show a circadian rhythm in %FMD in pre-menopausal women, which disappears after the menopause. The reduction in %FMD and an absence of a day-night variation in %FMD in postmenopausal women may have important implications for the incidence of coronary heart disease in women after the menopause. PMID- 16466870 TI - Investigating the moisture-induced crystallization kinetics of spray-dried lactose. AB - Gravimetric water sorption experiments were performed to study the crystallization behavior of amorphous spray-dried lactose over a wide range of temperature and humidity conditions. Experiments performed at 25 degrees C between 48 and 60% relative humidity (RH) showed that the onset time to crystallization increased dramatically with decreasing humidity. At 55% RH and above, crystallization occurred in a single detectable step, while below a two step process was observed. Experiments performed at 51% RH between 22 and 32 degrees C indicated the induction time to crystallization onset increased with decreasing temperature. Above 25 degrees C at 51% RH, crystallization occurred in one measurable step, while below crystallization occurred in two steps. The constant RH with varying temperature results were modeled to determine the crystallization mechanism. Above 25 degrees C a mechanism consisting of two competing reaction sequences fit the data with a 0.9997 correlation coefficient. Both reaction sequences have two steps: an auto-catalytic first step is followed by a three-dimensional diffusion controlled water loss step. PMID- 16466871 TI - Increased resistance to free radical damage induced by low-level sound conditioning. AB - Conditioning is the phenomenon where exposure to moderate-level acoustic stimuli can increase the ear's resistance to subsequent more intense sound exposures. In recent years, research has shown that conditioning increases the availability of antioxidant enzymes which presumably protects the ear from oxidative stress induced by a traumatic noise exposure [Jacono, A.A., Hu, B., Kopke, R.D., Henderson, D., Van De Water, T.R., Steinman, H.M., 1998. Changes in cochlear antioxidant enzyme activity after sound conditioning and noise exposure in the chinchilla. Hear Res 117, 31-8]. The current study was designed to assess whether the increase in endogenous antioxidants seen following conditioning could provide protection from oxidative stress induced by Paraquat, a potent generator of superoxide. Chinchillas were exposed to a conditioning noise, 500 Hz OBN at 95 dB for 6 h/day for 10 days, followed 5 days later with Paraquat application to the round window. Controls underwent the Paraquat application surgery, without prior conditioning. Evoked potential thresholds were determined prior to conditioning, at day 1, 5 and 10 during conditioning, at day 15 (5 days after conditioning), and at day 17, 19, 23, and 35 (1, 3, 7, and 20 days post-Paraquat). The conditioned animals showed reductions in permanent threshold shift and reduced inner hair cell loss relative to controls. These results reinforce the hypothesis that antioxidants are primary mediators of the conditioning effect. PMID- 16466872 TI - Expression of CLC-K chloride channels in the rat cochlea. AB - Current models of the lateral K+ recycling pathway in the mammalian cochlea include two multicellular transport networks separated from one another by three interstitial gaps. The first gap is between outer hair cells and Deiters cells, the second is between outer sulcus cells and type II spiral ligament fibrocytes and the third is between intermediate and marginal cells in the stria vascularis. K+ taken up by cells bordering these interstitial spaces is accompanied by Cl-. Maintaining appropriate electrolyte balance and membrane potentials in these cells requires a mechanism for exit of the resorbed Cl-. One possible candidate for regulating this Cl- efflux is ClC-K, a chloride channel previously thought to be kidney specific. Here, we demonstrate the expression of both known isoforms of ClC-K in the organ of Corti, spiral ligament and stria vascularis of the rat cochlea by immunohistochemical, Western blot and RT-PCR analysis. These results indicate a role for ClC-K in mediating Cl- recycling in the cochlea. The widespread expression of both ClC-K isoforms in the cochlea may help to explain the symptoms of Bartter's syndrome Type III, a mutation in the hClC-Kb gene (human homologue of ClC-K2), which results in renal salt wasting without deafness. These data support the hypothesis that both isoforms of ClC-K are co expressed in some cell membranes and account for the preservation of hearing in the presence of a mutation in only one channel isoform. PMID- 16466874 TI - Response to broadband repetitive stimuli in auditory cortex of the unanesthetized rat. AB - This study examines the ability of multi-unit clusters (MUCs) in layer IV/V of primary auditory cortex of the awake rat to respond to a series of broadband click trains. The data from 113 multi-unit clusters were analyzed for synchronous and nonsynchronized responses using several methods. Synchronous responses were measured using window analysis, circular statistics and spectral analysis. Nonsynchronous responses were measured during different time intervals during the click train (first 50 ms, 50-450 ms, and the entire click train). The results demonstrate that multi-unit clusters are capable of synchronizing to clicks at rates up to 166 Hz. The mean synchronization boundary (limiting rate) for the group was found to be 72 Hz. Mean peak response rate, mean response duration, and mean time-to-peak response decreased as the stimulus presentation rate (SPR) increased, resulting in a temporal sharpening of the population response. For fast SPRs (>50 Hz), 50% of MUCs exhibited nonsynchronous responses in which the firing rate increased with SPR, although this activity was most prevalent during the first 50 ms of the response. Sustained increases in firing rate with SPR were seen in 8% of the MUCs, while another 38% of MUCs exhibited sustained decreases during the click train. PMID- 16466875 TI - Horizontal and vertical effects of eye-position on sound localization. AB - The effect of gaze direction on the localization of sound sources was investigated in the azimuthal and elevational dimension using a pointing task. In both dimensions, eccentric eye-position induced a significant shift in sound localization that was opposite to the direction of eccentricity. This finding is in accordance with the view that the azimuthal and elevational components of the auditory spatial information are processed in common neural substrates. PMID- 16466873 TI - Uptake of fluorescent gentamicin by vertebrate sensory cells in vivo. AB - Aminoglycoside uptake in the inner ear remains poorly understood. We subcutaneously injected a fluorescently-conjugated aminoglycoside, gentamicin Texas Red (GTTR), to investigate the in vivo uptake of GTTR in the inner ear of several vertebrates, and in various murine sensory cells using confocal microscopy. In bullfrogs, GTTR uptake was prominent in mature hair cells, but not in immature hair cells. Avian hair cells accrued GTTR more rapidly at the base of the basilar papilla. GTTR was associated with the hair bundle; and, in guinea pigs and mice, somatic GTTR fluorescence was initially diffuse before punctate (endosomal) fluorescence could be observed. A baso-apical gradient of intracellular GTTR uptake in guinea pig cochleae could only be detected at early time points (<3h). In 21-28 day mice, cochlear GTTR uptake was greatly reduced compared to guinea pigs, 6-day-old mice, or mice treated with ethacrynic acid. In mice, GTTR was also rapidly taken up, and retained, in the kidney, dorsal root and trigeminal ganglia. In linguinal and vibrissal tissues rapid GTTR uptake cleared over a period of several days. The preferential uptake of GTTR by mature saccular, and proximal hair cells resembles the pattern of aminoglycoside-induced hair cell death in bullfrogs and chicks. Differences in the degree of GTTR uptake in hair cells of different species suggests variation in serum levels, clearance rates from serum, and/or the developmental and functional integrity of the blood labyrinth barrier. GTTR uptake by hair cells in vivo suggests that GTTR has potential to elucidate aminoglycoside transport mechanisms into the inner ear, and as a bio-tracer for in vivo pharmacokinetic studies. PMID- 16466876 TI - Effect of ethanol extract of Lepidium meyenii Walp. on osteoporosis in ovariectomized rat. AB - Maca (Lepidium meyenii Walp.) is a cruciferous plant from the Andes of Peru. The root of Maca is traditionally employed for its supposed properties in aphrodisiacs and improving fertility, it also has been widely used to help alleviate the symptoms of menopause. The purpose of this study was to evaluate the effect of ethanol extract of Maca on postmenopausal osteoporosis in ovariectomized rats. Female Sprague-Dawley rats were divided into four groups: Sham-operated and ovariectomized groups were fed with equivolume of distilled water, and the remaining ovariectomized groups were orally administrated with ethanol extract of Maca at 0.096 and 0.24 g/kg for 28 weeks. The findings derived from the basis of bone mineral density, biomechanical, biochemical and histopathological parameters indicated that higher dose of ethanol extract of Maca was effective in the prevention of estrogen deficient bone loss. PMID- 16466877 TI - Sensitivity of autopsy and radiological examination in detecting bone fractures in an animal model: implications for the assessment of fatal child physical abuse. AB - Skeletal injuries are often strong indicators of child abuse and their detection is therefore crucial. The aim of this study was to compare the sensitivity of three diagnostic approaches, namely autopsy, traditional (conventional) radiology, and computed tomography on "battered" piglets, in order to verify the sensitivity of each method, with respect to the true number of bone fractures assessed once the piglet was skeletonised (osteological control). Four newborn cadaver piglets who had died from natural causes were severely beaten post-mortem in every district of the body. Traditional radiography, computed tomography (CT) and autopsy were performed. The piglet was then macerated until skeletonised and the number of all fractures present recorded (osteological control). On the cranium, traditional radiology revealed only 35% circa of actual fractures, autopsy detected only 31% (P<0.01 for both comparisons versus osteological control), whereas CT imaging detected all fractures actually present. For ribs, radiology detected only 47% of all fractures present, and autopsy 65% circa (P>0.05 for both comparisons versus osteological control), while CT scans detected 34% (P<0.01). In suspected cases of fatal child abuse, we suggest that the bones of specific districts be directly analysed either at autopsy or by collecting specific diagnostic sites, such as parts of the rib cage, and subjecting them to maceration. The removed areas could be replaced with artificial material for cosmetic purposes. The authors stress the importance of combined radiological, CT scan, autopsy and osteological survey in the detection of perimortem bone fractures. PMID- 16466878 TI - Endolymphatic sac tumor located around semicircular canals. AB - We report a case of endolymphatic sac tumor (ELST). A 48-year-old female had recurrent and slowly progressive hearing loss, accompanied with dizziness like Meniere's disease. A tumor was located around the semicircular canals, and was detected on CT and MRI. The patient underwent total removal of the tumor using a transmastoid approach. Histopathological examinations agreed with features of an ELST. The tumor was highly suspected to have originated from the rugose portion of the endolymphatic sac or the endolymphatic duct, based on surgical and imaging studies. Structure of the membranous labyrinth was preserved regardless of the existence of the tumor around semicircular canals with bone destruction. ELSTs seem to have an osteolytic or osteophilic nature, by examining patterns of tumor infiltration. PMID- 16466879 TI - Paradigm shift in treatment of Wilson's disease: zinc therapy now treatment of choice. AB - Zinc therapy has replaced penicillamine as first-line therapy for Wilson's disease. New guidelines reflect the paradigm shift in treatment that has occurred in recent years. In the old paradigm, Wilson's disease was seen as genetic disorder associated with the accumulation of copper in the liver and in other organs once the liver had become overloaded with copper. When left untreated, the disease was regarded as uniformly fatal. The old treatment guidelines advised, 'decoppering' with penicillamine because this chelating agent was considered effective in restoring most patients to health. Before the start of treatment, patients were warned that their symptoms could worsen during the first weeks or months of therapy, so as to prevent them from abandoning penicillamine therapy in dismay. In the new paradigm, Wilson's disease is seen as a hereditary disorder associated with copper intoxication. The essence of symptomatic Wilson's disease is poisoning by free copper in the blood, that is, by copper that is not bound to ceruloplasmin. This form of copper is toxic, whereas accumulated copper and copper that is bound to ceruloplasmin or metallothionein is not. The treatment of symptomatic Wilson's disease is no longer aimed at 'decoppering', the removal of accumulated copper, but at the normalization of the free copper concentration in blood, to reverse the copper poisoning. This can be achieved safely and effectively with zinc therapy. Zinc induces metallothionein, a highly effective detoxification protein that binds copper. Oral zinc therapy leads to storage of metallothionein-bound copper in the mucosa of the gut and to the excretion of copper via the stools. New treatment guidelines advise against the use of chelating agents as initial treatment because they may aggravate copper intoxication and cause iatrogenic deterioration. PMID- 16466880 TI - ACTH therapy for Taiwanese children with West syndrome -- efficacy and impact on long-term prognosis. AB - To study the efficacy of adrenocorticotrophic hormone (ACTH) in treating Taiwanese children with West syndrome (WS) and the impact on long-term prognosis, 66 patients with WS (54 symptomatic and 12 cryptogenic) were collected from 1987 to 1998 in a medical center in Taiwan. A total of 53 patients were enrolled in this study and treated with ACTH at the dosage of 2.5IU/kg daily for 2 weeks with gradual tapering in subsequent 6 weeks. Immediate responses, side effects of ACTH and long-term outcomes of the patients including seizure and developmental status were evaluated during the average follow-up period of 35.6 months. The spasm-free percentage after one or two courses of ACTH treatment was 77.4%. Nine (17%) patients encountered severe side effects such as major infections, which prompted us to stop ACTH. At the end of follow-up, 22 (41.5%) patients had intractable seizures but 25 (47.2%) patients remained seizure free with or without anticonvulsants. The ACTH-responders had a better chance of remaining seizure free (P<0.05). Regarding the long-term developmental outcome, 12 (22.6%) patients had normal or borderline development; two thirds of them belonged to the crytpogenic group. Six (11.3%) patients expired and 24 (45.3%) were severely retarded; all but one of them belonged to the symptomatic group. The prognosis of WS heavily relies on whether a patient is cryptogenic or symptomatic (P<0.001). Good response to therapy or short treatment lag did not favorably affect the developmental outcomes of the symptomatic cases. We conclude that the long-term outcomes of WS in Taiwan were generally poor despite of treatment. Only cryptogenic patients had favorable prognosis. For symptomatic patients, ACTH therapy may be used to control the spasms and decrease the incidence of subsequent epilepsy, but it will not improve developmental outcome. Considering a high percentage of severe side effects in our study, a lower dosage of ACTH with adequate therapeutic efficacy but less side effects should be considered for treating Taiwanese children with WS. PMID- 16466882 TI - Effect of additional auditory and visual stimuli on continuous performance test (noise-generated CPT) in AD/HD children -- usefulness of noise-generated CPT. AB - The continuous performance test (CPT) is designed to measure sustained attention quantitatively. Several CPTs are used clinically. We have made changes to the conventional type of visual CPT, by displaying auditory and visual noise along with target or non-target stimuli. By influencing the recognition of the subjects in this way, the changes were intended to increase the sensitivity of detection of inattention and impulsiveness, to make CPT more useful for diagnosis, and to examine the effect of noise on AD/HD children during CPT performance. Its usefulness for AD/HD diagnosis and the reaction of AD/HD children to noise were examined using newly developed computer software. Using this CPT analysis, a significant difference was observed in all measurements, except mean reaction time, between the control and AD/HD groups, showing that it was useful as a supplementary diagnostic method for AD/HD, and was more useful in the younger age group than in the older age group, as the same for conventional CPTs. As compared to no-noise sessions, commission and omission errors both increased significantly in auditory and visual noise sessions. Thus, analyzing the changes in measurements during noise sessions will improve the diagnosis of inattention and combined AD/HD subtypes. Furthermore, it was suggested that analysis of the effects of noise on AD/HD children will benefit their handling in an educational environment. Since omission errors were decreased in AD/HD children by noise during the CPT performance as compared to the control group, noise may induce attention in AD/HD children. The present study presents new findings on the responses to noise of AD/HD children during the CPT. PMID- 16466883 TI - Changes in cerebral and ocular hemodynamics in Behcet's disease assessed by color coded duplex sonography. AB - AIM: To quantify the cerebral and retrobulbar hemodynamics in Behcet's disease with and without ocular involvement and compared with that of healthy controls. MATERIALS AND METHODS: Of 51 people studied, 17 had Behcet's disease with ocular involvement, 17 had Behcet's disease without ocular involvement, and 17 were healthy controls. A single eye was examined in each patient. Peak systolic velocity (PSV), end-diastolic velocity (EDV), time-averaged maximum velocity (Tamax), and resistance index (RI) were evaluated in the ophthalmic (OA), posterior ciliary (PCA), central retinal (CRA) and middle cerebral artery (MCA). Additionally, the average blood flow velocities in the central retinal vein (CRV), and acceleration time (AT) and pulsatility index (PI) in the MCA were calculated. RESULTS: The mean EDV in the PCA was 25% lower and RI was higher in patients with ocular involvement of BD than in patients without involvement (p = 0.006 and p = 0.005, respectively) and in healthy controls (p = 0.003 and p = 0.004, respectively). Differences were smaller in comparisons of the CRA and absent on comparisons of the OA and MCA. The acceleration time of the MCA was significantly higher in patients with Behcet's disease than in healthy controls (p = 0.03). CONCLUSION: This study suggests that the flow hemodynamics in retrobulbar circulation has more altered Behcet's disease with ocular involvement than without ocular involvement and healthy control. Additionally, the cerebral hemodynamic might be affected in patients with Behcet's disease compared with healthy controls. PMID- 16466884 TI - Exploration of Amycolatopsis diversity in soil using genus-specific primers and novel selective media. AB - Large numbers of actinomycetes provisionally assigned to the genus Amycolatopsis were isolated from soil samples using a dilution plate procedure and three novel media designed to be selective for members of this genus. A set of genus-specific oligonucleotide primers developed for the rapid identification of unknown Amycolatopsis strains was used to determine whether representative strains taken from the selective isolation plates produced the diagnostic amplification product. The 175 isolates which tested positive were assigned to major, minor and single-membered colour-groups. Representatives of the major colour-groups had a profile of chemical and morphological properties consistent with their assignment to the genus Amycolatopsis; strains taken to represent most of these taxa formed distinct phyletic lines in the Amycolatopsis 16S rRNA gene tree. Many of the representative isolates were closely related to either Amycolatopsis mediterranei or to Amycolatopsis orientalis or formed new evolutionary lines in either the Amycolatopsis orientalis or Amycolatopsis methanolica 16S rRNA subclades. It is evident that the genus Amycolatopsis is grossly underspeciated. PMID- 16466885 TI - Assay of amikacin in the skin by high-performance liquid chromatography. AB - Amikacin is used in the systemic treatment of serious infections, but also locally for the treatment of skin infections. The aim of this work was to develop and validate a simple procedure for amikacin determination inside the epidermal tissue: this implies a simple method for an efficient drug extraction from the skin and a clean and easy HPLC analysis. Amikacin was extracted from epidermis samples with 500 microl of a mixture methanol-water-0.05 M NaOH (5:5:2 v/v/v) at 60 degrees C for 1 h. After filtration, the obtained solution was derivatized (1 fluoro-2,4-dinitrobenzene at 90 degrees C for 10 min) and analyzed by HPLC, on a C18 microBondapack 300 mmx4.6 mm column thermostatted at 45 degrees C. The mobile phase was a mixture of acetonitrile-water-acetic acid (47:53:0.1 v/v/v) at a flow rate of 1.5 ml/min and the UV detector was set at 365 nm. The derivatization and HPLC analysis were validated in the concentration interval 1.64-49.21 microg/ml. The linearity resulted very good (R=0.9995); the R.S.D.% varied between 0.20% and 3.89% depending on the concentration and the ER% was included between 5.4 and 0.9. The extraction method used demonstrated to be specific and the recovery resulted about 93%. The extraction, derivatization and HPLC assay has good reproducibility, sensitivity and specificity resulting in a reliable method for biopharmaceutical studies of AK distribution in the epidermis. PMID- 16466886 TI - Stationary phases in the screening of drug/impurity profiles and in their separation method development: identification of columns with different and similar selectivities. AB - The classification or characterization of stationary phases based on chromatographic parameters, in general, requires different test solutes/mixtures and several mobile phases. To simplify the classification/characterization of reversed-phase liquid chromatographic columns, to be used in separating drug/impurity profiles, a new test procedure was proposed. It consists of injecting two mixtures of relatively similar active substances applying a standard gradient. The aim was to evaluate from this approach the selectivity differences and overall separation quality of newly tested columns compared to that in an earlier selected set of eight stationary phases. The selectivity differences of the columns were evaluated by correlation coefficient-based weighted-average-linkage dendrograms and color maps. Derringer's desirability functions were used to rank similar stationary phases according to their overall separation quality. Four columns of 27 examined were, for instance, considered different from the earlier selected eight and could be added to the selection. A number of tested stationary phases might be considered as alternatives for some from the initial set. For three columns the newly tested stationary phases did not contain alternatives. PMID- 16466887 TI - A case of Mycobacterium nonchromogenicum pulmonary infection showing multiple nodular shadows in an immunocompetent patient. AB - Mycobacterium nonchromogenicum is generally considered nonpathogenic. However, M. nonchromogenicum rarely causes human disease; particularly, pulmonary disease is extremely rare. The common finding of M. nonchromogenicum pulmonary infection on chest X-ray is a solitary cavity. The present report describes an unusual case of M. nonchromogenicum primary pulmonary infection showing multiple nodular shadows. PMID- 16466888 TI - Prevalence of Helicobacter pylori infection in children (south of Iran). AB - The prevalence of Helicobacter pylori in children ranges from 10% to more than 80%. High prevalence occurs in developing countries, and after colonization it takes an extended period to be eradicated by the immune system. To evaluate the prevalence and age distribution of H. pylori infection in children in Shiraz (a city in the south of Iran), we collected 593 stool samples from children selected randomly from 5 age groups. Infection was determined based on antigen immunoassay in stool using the enzyme-linked immunosorbent assay method. The prevalence rates were 82%, 98%, 88%, 89%, and 57% in age groups of 9 months, and 2, 6, 10, and 15 years, respectively. There were no significant differences between the prevalence of H. pylori infection in the first 4 age groups (P > 0.05), but there was a significant decrease in the 15-year-old group (P < 0.05). The prevalence of H. pylori infection in the south of Iran is very high. The infection begins at infancy and remains high until late childhood. PMID- 16466889 TI - Urine polymerase chain reaction is not as sensitive as urine antigen for the diagnosis of disseminated histoplasmosis. AB - We developed a colorimetric microtiter plate polymerase chain reaction enzyme immunoassay (PCR-EIA) for the detection of Histoplasma capsulatum in urine. The specificity of the PCR assay was confirmed using H. capsulatum (positive control) and Blastomyces dermatitidis (negative control) isolates. The analytical sensitivity of the PCR assay was determined by testing urine samples spiked with freshly grown H. capsulatum organisms and was 2 yeasts per reaction in urine and 0.2 yeasts per reaction in urine sediment after centrifugation. Fifty-one urine specimens positive for H. capsulatum antigen and 25 urine specimens from healthy volunteers were tested blindly. Patient specimens also were cultured for H. capsulatum. The PCR assay was positive in 4 (7.8%) of 51 urine specimens containing antigen and negative in urine specimens from healthy volunteers. The positive PCR results occurred in 4 of 5 urine specimens that were positive by culture, and each exhibited high level of antigenuria (>20 U). Urine cultures were not positive in 24 urine specimens with an antigenuria of 1-19.9 U, but were positive in 5 of 27 urine specimens with antigenuria >20 U. Thus, positive PCR results in urine specimens correlate with positive culture results, but not with antigenuria. The low sensitivity of this PCR assay in urine limits its use in the diagnosis of disseminated histoplasmosis. PMID- 16466890 TI - Clonal occurrences of multidrug-resistant Gram-negative bacilli: report from the Meropenem Yearly Susceptibility Test Information Collection Surveillance Program in the United States (2004). AB - The Meropenem Yearly Susceptibility Test Information Collection Program is a global, longitudinal resistance surveillance network of more than 100 medical centers worldwide monitoring the susceptibility of bacterial pathogens to carbapenems and other broad-spectrum agents. Between 1999 and 2004, a total of 10 16 US medical centers referred up to 200 nonduplicate isolates from clinical infections to a central processing laboratory. Over this 6-year period, the antimicrobial activity of 12 broad-spectrum agents was assessed against 15990 bacterial isolates using Clinical and Laboratory Standards Institute (formerly the National Committee for Clinical Laboratory Standards) reference methods. Analysis of the MIC results by year within organism species and/or organism groups revealed a continued decrease in fluoroquinolone susceptibility (ciprofloxacin and levofloxacin), especially among indole-positive Proteae ( 22.1%), Escherichia coli (-17.0%), Enterobacter species (-10.0%), and Proteus mirabilis (-7.6%) isolates. Antibiogram analysis of strains demonstrating multidrug resistance from the same institutions were further characterized by automated ribotyping and pulsed-field gel electrophoresis to determine clonality. In 2004, a total of 165 selected multidrug-resistant (MDR) strains produced 64 different ribotypes with the largest representing 31 Escherichia coli isolates from 6 medical centers. Other clusters were also identified within single medical centers among Enterobacter cloacae (6 strains), Providencia stuartii (5 strains), and Morganella morganii (4 strains). A significant clonal outbreak encompassing 40 Acinetobacter baumannii isolates from 3 centers in a single endemic region was identified in 1999 and has persisted through 2004. Continued surveillance of these broad-spectrum antimicrobial agents against MDR pathogens appears warranted to monitor the incidence and spread of resistant clones causing serious infections. Possible emergence of resistance mechanisms via clonal dissemination proves to be among the principle threats that compromise carbapenem therapy, where meropenem maintains the broadest spectrum of coverage. PMID- 16466891 TI - Prospective evaluation of the treatment and outcome of community-acquired pneumonia according to the Pneumonia Severity Index in VHA hospitals. AB - The objective of the study were to determine if nationally recognized community acquired pneumonia (CAP) guidelines (specific to antibiotic therapy) were being followed and to identify outcomes of treatment in hospitals that are VHA members. This was a prospective study using a medication use evaluation in an inpatient setting conducted in 46 institutions in the United States during the 1998-1999 CAP season. The subjects were 875 adult patients (> or =18 years of age) admitted from the emergency department or ambulatory care setting with a chest X-ray confirmed diagnosis of CAP. Treatment pathways were in place in 58.7% (27/46) of institutions, with 18.3% of patients treated according to pathways. Twenty-seven percent of patients were PSI class I or II. A pathogen (blood or sputum) was identified in <10% of patients. The first dose of antibiotic was administered to patients 65% of the time in the emergency department. Antibiotic therapy in 592 of the 694 admitted to a general medical unit (mortality rate, 3%) complied with 1998 Infectious Diseases Society of America (IDSA) guidelines compared with 26 of the 65 admitted to the intensive care unit (ICU) (mortality rate, 4.6%). In patients admitted to other nongeneral medical, non-ICU areas, IDSA guidelines were followed in 95% of the patients. Mean length of stay and mortality for PSI classes I-V were 4.5, 4.6, 6.9, 6.2, and 7.1 days, respectively, and 0%, 0.7%, 1.1%, 2.5%, and 10.5%, respectively. Antibiotic therapy was modified in 733 of 875 patients. Approximately 90% of patients were eligible for conversion to oral (per os) therapy before discontinuation of parenteral (intravenous) antibiotics (mean time to eligibility, 1.8 days of parenteral antibiotics), with conversion in 65% (mean time to conversion to oral therapy, 4.6 days). Resolution of CAP occurred in 92% of patients; deterioration was more common in PSI class IV and V patients. In conclusion, inhospital mortality rates for all PSI classes were similar to those found in other recently conducted studies despite limited adherence to pathways. Greater use of treatment guidelines for patients admitted to the ICU and awareness of the intravenous to per os antibiotic conversion process are suggested. PMID- 16466892 TI - Susceptibility studies of piperazinyl-cross-linked fluoroquinolone dimers against test strains of Gram-positive and Gram-negative bacteria. AB - Susceptibility testing was used to evaluate potential spectrum of action for piperazinyl-cross-linked fluoroquinolone dimers against test strains of Streptococcus pneumoniae, Streptococcus pyogenes, Escherichia coli, Pseudomonas aeruginosa, Mycobacterium tuberculosis, and vancomycin-resistant Enterococcus faecium (VRE) and to evaluate dimers against fluoroquinolone-resistant and fluoroquinolone-susceptible strains of streptococci. Individual dimers displayed equivalent or lowered MIC values compared with parent fluoroquinolone monomers against test strains of S. pneumoniae, S. pyogenes, E. coli, and VRE. Raised MIC values were observed for all dimers in comparison to monomers against test strains of P. aeruginosa and E. coli. In comparison to parent fluoroquinolones, all dimers displayed decreased percent inhibition of growth against M. tuberculosis. Structural requirements for activity of dimers and partial dimers against all organisms, including lower MICs against certain fluoroquinolone resistant and fluoroquinolone-susceptible strains of streptococci, were consistent with requirements previously observed for dimers against fluoroquinolone-susceptible and fluoroquinolone-resistant strains of S. aureus. In contrast, the 10- to 100-fold lowering of MICs against wild-type and fluoroquinolone-resistant strains of S. aureus previously observed for individual cross-linked dimers was not observed with test strains of the various organisms used here. PMID- 16466893 TI - Salmonella group C necrotizing fasciitis: a case report and review of the literature. AB - Localized salmonella soft tissue infections have been rarely described in humans. We report a case of necrotizing fasciitis caused by Salmonella serogroup C in a patient with systemic lupus erythematosus who was successfully treated with surgical debridement and cefoperazone-sulbactam. In addition, we provide a literature review on risk factors and treatment of this uncommon disease sequelae. PMID- 16466894 TI - Lack of association of Staphylococcus aureus type A beta-lactamase with cefazolin combined with antimicrobial spacer placement prosthetic joint infection treatment failure. AB - Association of cefazolin treatment failure with type A beta-lactamase-producing Staphylococcus aureus has been suggested. The prevalence of beta-lactamase gene types among 23 methicillin-susceptible S. aureus (MSSA) isolates associated with prosthetic joint infection (PJI) treated with cefazolin was determined using polymerase chain reaction (PCR), and clinical and microbiologic outcomes were assessed. PCR revealed 4 isolates without blaZ, and 12 with type A, 2 with type B, and 5 with type C blaZ. Of 13 patients undergoing resection arthroplasty with subsequent reimplantation, all received antimicrobial spacer placement at resection with or without antimicrobial-impregnated polymethylmethacrylate at reimplantation. All 13 cases had tissue cultures at time of reimplantation that were negative for S. aureus and 11 had histopathology specimens showing no acute inflammation. Of 8 type A cases undergoing reimplantation, all prostheses remained in place at follow-up (median, 798 days; range, 32-927 days). We conclude that type A blaZ is common in MSSA PJI and that cefazolin therapy for blaZ MSSA PJI can be successful when combined with 2-stage reimplantation and local antimicrobial therapy. PMID- 16466895 TI - A simple and low-cost paper-bridged method for Salmonella phase reversal. AB - We have developed a simple, rapid, and low-cost "paper-bridged" method for Salmonella phase reversal. More than 3500 isolates were tested in our laboratory, and the results indicated that this paper-bridged method is a useful alternative for phase reversal. PMID- 16466896 TI - Detection of the reemerging agent Burkholderia mallei in a recent outbreak of glanders in the United Arab Emirates by a newly developed fliP-based polymerase chain reaction assay. AB - A polymerase chain reaction (PCR) assay targeting the flagellin P (fliP)-I S407A genomic region of Burkholderia mallei was developed for the specific detection of this organism in pure cultures and clinical samples from a recent outbreak of equine glanders. Primers deduced from the known fliP-IS407A sequence of B. mallei American Type Culture Collection (ATCC) 23344(T) allowed the specific amplification of a 989-bp fragment from each of the 20 B. mallei strains investigated, whereas other closely related organisms tested negative. The detection limit of the assay was 10 fg for purified DNA of B. mallei ATCC 23344(T). B. mallei DNA was also amplified from various tissues of horses with a generalized B. mallei infection. The developed PCR assay can be used as a simple and rapid tool for the specific and sensitive detection of B. mallei in clinical samples. PMID- 16466897 TI - Decreased susceptibility to fluoroquinolones and gyrA gene mutation in the Salmonella enterica serovar Typhi and Paratyphi A isolated in Katmandu, Nepal, in 2003. AB - Typhoid fever is the most common clinical diagnosis among febrile patients presenting to hospital in Katmandu. Salmonella enterica serovar Typhi (S. enterica serovar Typhi) and Salmonella enterica serovar Paratyphi A (S. enterica serovar Paratyphi A) with decreased susceptibility to fluoroquinolones and resistance to nalidixic acid are common in recent years. In the present study, we examined the in vitro susceptibility to fluoroquinolones and the presence of gyrA gene mutations in 30 clinical strains of S. Typhi and 39 of S. Paratyphi A, all of which were isolated in Katmandu, Nepal, in 2003. In those strains, we found that 73.3% and 94.9% of S. Typhi and S. Paratyphi A strains contained gyrA gene mutation, and showed the resistance to a quinolone, nalidixic acid, and decreased susceptibility to fluoroquinolones, ciprofloxacin, and levofloxacin. Although fluoroquinolones may still be useful as antibiotics for the treatment of typhoid fever, clinicians should be aware of the possibility of treatment failures of infections with S. Typhi and S. Paratyphi A strains with decreased susceptibility to fluoroquinolones. PMID- 16466898 TI - Cost savings with implementation of PNA FISH testing for identification of Candida albicans in blood cultures. AB - Antifungal expenditures are substantial for many hospitals. Using caspofungin for the treatment of candidemia accounts for a sizable proportion of the costs. A cost minimization study that used a decision analytic model was done to compare in-hospital diagnosis and treatment costs using the Candida albicans peptide nucleic acid fluorescence in situ hybridization (PNA FISH) test versus the C. albicans screen test for differentiating C. albicans from non-albicans Candida species bloodstream infections. Assuming physician notification of yeast identity concurrent with blood culture positivity, potential savings resulting from use of the C. albicans PNA FISH test compared with the C. albicans screen test averaged $1837 per patient treated, although laboratory costs for doing the C. albicans PNA FISH test ($82.72) exceeded those for the C. albicans screen test ($2.83). Savings were realized through a decrease in antifungal drug costs, particularly caspofungin. Incorporating the C. albicans PNA FISH test as part of the initial identification algorithm for yeasts recovered from blood can result in substantial savings for hospitals. PMID- 16466899 TI - Viability of Leptospira in BacT/ALERT MB media. AB - Recovery of Leptospira in the clinical setting is typically low as specialized culture media is needed. Previous data demonstrated that blood culture media commonly available to most clinical laboratories do not adequately sustain viable Leptospira. We hypothesized that mycobacterial blood culture medium, which is often readily available to most clinical laboratories, might be able to support the growth of Leptospira. Leptospires and fresh human blood were inoculated into BacT/ALERT (bioMerieux, Durham NC) mycobacterial (MB) and enriched mycobacterial bottles. Standard aerobic (FA) and anaerobic (SN) bottles were also inoculated as a control group. Inoculated bottles were then evaluated for their ability to support Leptospira growth using dark-field microscopy, subculture, and an automated growth detection system. Viable leptospires were detected in MB bottles up to day 14. FA and SN were performed in accordance with prior data. We conclude that MB and enriched MB bottles of the BacT/ALERT blood culture system can support viable leptospires. PMID- 16466900 TI - Rapid identification of fungal pathogens in BacT/ALERT, BACTEC, and BBL MGIT media using polymerase chain reaction and DNA sequencing of the internal transcribed spacer regions. AB - We report a direct polymerase chain reaction/sequence (d-PCRS)-based method for the rapid identification of clinically significant fungi from 5 different types of commercial broth enrichment media inoculated with clinical specimens. Media including BacT/ALERT FA (BioMerieux, Marcy l'Etoile, France) (n = 87), BACTEC Plus Aerobic/F (Becton Dickinson, Microbiology Systems, Sparks, MD) (n = 16), BACTEC Peds Plus/F (Becton Dickinson) (n = 15), BACTEC Lytic/10 Anaerobic/F (Becton Dickinson) (n = 11) bottles, and BBL MGIT (Becton Dickinson) (n = 11) were inoculated with specimens from 138 patients. A universal DNA extraction method was used combining a novel pretreatment step to remove PCR inhibitors with a column-based DNA extraction kit. Target sequences in the noncoding internal transcribed spacer regions of the rRNA gene were amplified by PCR and sequenced using a rapid (24 h) automated capillary electrophoresis system. Using sequence alignment software, fungi were identified by sequence similarity with sequences derived from isolates identified by upper-level reference laboratories or isolates defined as ex-type strains. We identified Candida albicans (n = 14), Candida parapsilosis (n = 8), Candida glabrata (n = 7), Candida krusei (n = 2), Scedosporium prolificans (n = 4), and 1 each of Candida orthopsilosis, Candida dubliniensis, Candida kefyr, Candida tropicalis, Candida guilliermondii, Saccharomyces cerevisiae, Cryptococcus neoformans, Aspergillus fumigatus, Histoplasma capsulatum, and Malassezia pachydermatis by d-PCRS analysis. All d PCRS identifications from positive broths were in agreement with the final species identification of the isolates grown from subculture. Earlier identification of fungi using d-PCRS may facilitate prompt and more appropriate antifungal therapy. PMID- 16466901 TI - Temporal response of liver signal transduction elements during in vivo endotoxin challenge in cattle: effects of growth hormone treatment. AB - We quantified the changes in abundance of inducible nitric oxide synthase (iNOS) and associated tissue signal transduction pathway elements (STPEs) in the bovine liver in response to lipopolysaccharide (LPS) challenge and further assessed the impact on the LPS-driven variable responses as affected by daily treatment with recombinant growth hormone (GH) prior to LPS challenge. Twenty-four cross-bred beef steers were divided into GH-treated (recombinant bovine GH, Monsanto Inc., St. Louis, MO; 0.1mg/kg BW, i.m., daily for 12 days) and non-GH-treatment (control) groups (n=12/group). Liver biopsy samples were obtained from all animals at 0, 3, 6, and 24h after LPS challenge (E. coli 055:B5, 2.5 microg/kg BW, i.v. bolus) for Western blot analyses of iNOS and STPEs. In response to LPS, tissue levels of iNOS increased significantly (P<0.001) in the first 3h and persisted at levels greater than those at time 0 until 24h. GH further augmented levels of iNOS at 0, 3, and 6h resulting in an overall significant increase in the iNOS protein level (P<0.01). AKT/protein kinase B (AKT/PKB) phosphorylation levels at time 0 were not different between GH-treated and control animals; LPS increased the phosphorylation of AKT/PKB with GH treatment stimulating a four fold further increase of AKT/PKB phosphorylation. Effects similar to those on AKT/PKB were also observed on signal transducer and activator of transcription 5b (STAT5b). The family of mitogen-activated protein kinase (MAPK) showed different pattern of response. ERK1/2 phosphorylation increased 3h after LPS challenge but only in GH-treated group (P<0.01). Compared to 0 h, SAPK/JUN phosphorylation increased in both experimental groups 3, 6h (P<0.01), and 24h (P<0.05) after LPS. However, at 3h the increase was greater (P<0.01) in GH-treated than in control animals. No effect of LPS challenge or GH treatment on p38(MAPK) was observed. These results suggest that GH treatment has a significant impact on the differential activation of STPEs in the clinical response to LPS. PMID- 16466902 TI - Purification and characterization of feline ghrelin and its possible role. AB - Ghrelin, a novel 28-amino acid peptide with an n-octanoyl modification at Ser3, has been isolated from rat and human stomach as an endogenous ligand for the growth hormone secretagogue receptor. Here, we purified feline ghrelin and examined its possible physiological role in cats. The major active form of feline ghrelin is a 28-amino acid peptide octanoylated (C8:0) at Ser3; except for one amino acid residue replacement, this structure is identical to those of rat and human ghrelins. However, much structural divergence in peptide length and fatty acid modification was observed in feline ghrelin: peptides consisting of 27 or 26 amino acids lacking Gln14 and/or Arg28 were found, and the third serine residue was modified by octanoic acid (C8:0), decanoic acid (10:0), or unsaturated fatty acids (C8:1, C10:1 and C10:2). In agreement with the structural divergence, two kinds of cDNA with different lengths were isolated. Administration of synthetic rat ghrelin increased plasma growth hormone levels in cats, with a potency similar to that in rat or human. Plasma levels of ghrelin in cats increased approximately 2.5-fold after fasting. The present study indicates the existence of structural divergence in feline ghrelin and suggests that, as in other animals, ghrelin may play important roles in GH release and feeding in cats. PMID- 16466903 TI - Breast cancer sagittal/horizontal plane location influences axillary lymph node involvement. AB - AIM: To assess the influence of tumour location on axillary lymph node involvement (ALNI) and prognosis in breast cancer by evaluating the significance of the sagittal/horizontal alignment. METHODS: We compared 57 patients with superficially located breast carcinomas up to 3.0 cm with patients having lesions in posterior planes of the breast. Both groups were matched according to age, time of diagnosis, tumour size, grade, hormonal receptor status and tumour site within the frontal plane. Histologic evidence of skin involvement, excluding tumours fulfilling the criteria for pT4b, was defined as inclusion criteria and reference plane for superficial tumour location. RESULTS: Tumours situated in the superficial region of the breast, compared to those located in deeper planes, have an increased risk of ALNI (p=0.023), whereas no difference was observed with reference to disease-specific survival (p=0.203). CONCLUSION: This study shows that ALNI is dependent on sagittal/horizontal as well as frontal tumour location. Clinicians should be aware that tumours lying posteriorly may be at increased risk of occult spread outside axillary lymph nodes. PMID- 16466904 TI - Anxiety sensitivity and its dimensions across the anxiety disorders. AB - Anxiety sensitivity (AS) refers to the fear of anxiety-related sensations, which is thought to arise from beliefs about their harmful consequences. AS is a multidimensional construct that consists of fears of somatic, social, and cognitive aspects of anxiety. In the present study, we examined the relationship between AS dimensions, assessed by factor-analytically derived subscales of the Anxiety Sensitivity Index-Revised (ASI-R), and anxiety-related psychopathology in a sample of 232 treatment-seeking patients with anxiety disorders. Correlational analyses and comparisons among anxiety disorder patient groups and undergraduate students revealed a specific pattern of relationships between ASI-R subscale scores and anxiety-related psychopathology. In contrast, ASI-R total scores evidenced less discriminant validity. Implications for theoretical models of anxiety and directions for future research are discussed. PMID- 16466905 TI - NDPK-A (but not NDPK-B) and AMPK alpha1 (but not AMPK alpha2) bind the cystic fibrosis transmembrane conductance regulator in epithelial cell membranes. AB - Cystic fibrosis (CF) results from mutations within the cystic fibrosis transmembrane-conductance regulator (CFTR) protein. The AMP-activated protein kinase (AMPK) is a heterotrimer composed of different isoforms of the alphabetagamma subunits, where the alpha1 catalytic subunit binds CFTR. Nucleoside diphosphate kinase (NDPK, NM23/awd) converts nucleoside diphosphates to nucleoside triphosphates but also acts as a protein kinase. We recently showed that AMPK alpha1 binds NDPK-A in lung epithelial cytosol. Here we report that in the plasma membrane of human airway epithelial cells, NDPK-A and AMPK alpha1 associate with the plasma membrane via CFTR. We show that the regulatory domain of CFTR binds NDPK-A whereas AMPK gamma1 or gamma2 bind the first nucleotide binding domain (NBD1) and AMPK alpha1 binds the second (NBD2) of CFTR. We also show that NDPK-A specifically binds AMPK alpha1 and AMPK gamma2 subunits, thereby specifying the isozyme of AMPK heterotrimer that associates with CFTR at the membrane. Thus, the combined data provide novel insight into the subunit composition of the epithelial CFTR/AMPK/NDPK complex, such that: CFTR interacts specifically with AMPK alpha1, gamma2 and NDPK-A and not NDPK-B or AMPK gamma1. PMID- 16466906 TI - Effects in vitro and in vivo of a gap junction blocker on epileptiform activities in a genetic model of absence epilepsy. AB - We investigated the effects of carbenoxolone (CBX), a gap junctions (GJ) blocker, on epileptiform activities in vivo and in vitro. In a first series of experiments, i.p. CBX decreased the cumulative duration of cortical spike-wave discharges (SWD) in adult Genetic Absence Epilepsy Rats from Strasbourg (GAERS) without reduction in the SW amplitude or frequency. Since SWD are generated in thalamocortical networks, we studied the effect of CBX on thalamic and cortical activities elicited by 4-aminopyridine (4AP) in thalamocortical slices from GAERS or non-epileptic rats (NER). Spontaneous ictal-like activities (ILA) were recorded simultaneously in thalamus and somatosensory cortex. However, experiments where these structures were surgically separated showed that ILA were generated in the cortex and recorded by volume conduction in the thalamus. GABA dependent negative field potentials were also recorded in the cortex, either isolated or initiating ILA. After bath-applying CBX (100 microM), the frequency and cumulative duration of ILA decreased but less rapidly in GAERS than in NER slices and they disappeared at a time point when GABA-dependent negative potentials remained. These data suggest that GJ do not mediate the 4AP induced interneuronal synchronisation but may be implicated in the spreading of the synchronised activities from interneuronal networks to principal neurones. Our results show that CBX exerts an antiepileptic action in vivo, and that GJ blockers limits spread of synchronised activities in vitro. They may represent an appropriate target for development of new antiepileptic drugs. PMID- 16466907 TI - Reeler mutant mice exhibit seizures during recovery from isoflurane-induced anesthesia. AB - Reeler mice are a model of cortical malformation with enhanced seizure susceptibility. Data suggest that the propensity to anesthesia-induced seizures may be enhanced in animal models with developmental anomalies. Consequently, reeler mice were monitored behaviorally before, during and after isoflurane anesthesia. During recovery, 12% of reeler homozygotes had class I/II seizures while the remaining 88% exhibited convulsive seizures entailing opisthotonus and forepaw drumming. Similar behavior was not observed in controls. These data reveal that reeler mice display isoflurane-induced seizures and provide support for the hypothesis that developmental anomalies may predispose the central nervous system to anesthesia-induced seizures. PMID- 16466908 TI - Self-assembly of an amphiphilic derivative of chitosan and micellar solubilization of puerarin. AB - A kind of amphiphilic derivatives of chitosan (2-hydroxyl-3-butoxyl) propylcarboxymethyl-chitosan (HBP-CMCHS), has been synthesized, and the critical micelle concentration (cmc) of HBP-CMCHS was detected by the fluorescence method. The puerarin-loaded HBP-CMCHS micellar system was prepared by physical entrapped method. Result showed that when adding the same amount of puerarin, the solubilizing capacity was enhanced by increasing the concentration of HBP-CMCHS and temperature. Puerarin-loaded micellar system of HBP-CMCHS was characterized by TEM and DLS. TEM photograph revealed that the micelles were spherical and puerarin was solubilized in the cores of the spherical polymeric micelles. DLS showed that after solubilization the size of the micelles became bigger. In vitro tests showed that puerarin was slowly released from micellar solution and the release lasted up to 60 h by means of the dialysis method. PMID- 16466909 TI - Electrochemical behaviors of adrenaline at acetylene black electrode in the presence of sodium dodecyl sulfate. AB - The electrochemical behaviors of adrenaline at the acetylene black electrode in the presence of sodium dodecyl sulfate (SDS) were investigated by cyclic voltammetry and electrochemical impedance spectroscopy (EIS). The results indicated that the electrochemical responses of adrenaline were apparently improved by SDS, due to the enhanced accumulation of protonated adrenaline via electrostatic interaction with negatively charged SDS at the hydrophobic electrode surface. This was verified by the influences of different kinds of surfactants on the electrochemical signals of adrenaline. The electrochemical parameters of the adrenaline oxidation were explored by chronocoulometry. Under optimal working conditions, the oxidation peak current at 0.57 V was proportional to adrenaline concentration in the range of 5.0x10(-8) to 7.0x10(-6) mol/L, with a low detection limit of 1.0x10(-8)mol/L for 70s accumulation by differential pulse voltammetry (DPV). This method was applied to determine adrenaline in the hydrochloride injection sample. The results are satisfying compared with that by the standardized method of high performance liquid chromatography (HPLC). PMID- 16466910 TI - Kinetic studies of the interaction of fatty acids with phosphatidylcholine vesicles (liposomes). AB - The kinetics of addition of fatty acids (as alkaline solutions of the fatty acid anions) to pre-existing unilamellar phospholipid vesicles (mean diameter 100 nm) has been studied. The phospholipid DMPC (1,2-dimyristoyl-sn-glycero-3 phosphocholine) has been mainly used, together with three fatty acids, oleic acid (cis-9-octadecenoic acid), linoleic acid (cis,cis-9,12-octadecadienoic acid) and capric acid (decanoic acid). Experiments were performed above as well as below the main phase transition temperature (Tm) of DMPC vesicles. The pH chosen to study the fatty acid vesicle interaction (after fatty acid and vesicle mixing) was 8.5 in the case of oleic acid and linoleic acid and 7.4 for capric acid. In the absence of any pre-existing phospholipid vesicles, the addition of alkaline solutions of the fatty acid anions to corresponding buffer solutions of pH 8.5 or 7.4 leads to a partial protonation of the fatty acid anions again resulting in the formation of fatty acid vesicles. This process is rather slow, taking place over a period of hours/days, and the vesicles formed are very polydisperse and include a range of vesicle sizes/shapes. However, in the presence of pre-existing phospholipid vesicles the added fatty acids equilibrate readily within a few minutes and the size of the vesicles that form are then closely related to the size of the originally present phospholipid vesicles; the vesicles formed being generally somewhat larger than the pre-existing vesicles. In the case of the phospholipid DMPC, the mixed fatty acid/phospholipid vesicle system is often formed rather rapidly (particularly above Tm), so that stopped-flow methods have been applied to follow the kinetics of the process. It is proposed that most of the fatty acid molecules are initially rapidly incorporated into the bilayers of the pre-exisiting phospholipid vesicles as monomers, rather than that the added fatty acids form separate fatty acid vesicles. The mean vesicle sizes formed in the systems investigated have been analysed by using dynamic light scattering measurements. The behaviour of the DMPC system was found to be slightly different from the POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) system studied before, but the results are consistent with a model that involves growth and subsequent fission of the mixed vesicles. The study provides further support of the "matrix effect" in this type of system [S. Lonchin, P.L. Luisi, P. Walde, B.H. Robinson, J. Phys. Chem. B 103 (1999) 10910-10916]. The pre-existing DMPC vesicles act as a kind of seed to control the behavior of the system in the presence of added fatty acid anions. PMID- 16466912 TI - Screening genetically modified organisms using multiplex-PCR coupled with oligonucleotide microarray. AB - In this research, we developed a multiplex polymerase chain reaction (multiplex PCR) coupled with a DNA microarray system simultaneously aiming at many targets in a consecutive reaction to detect a genetically modified organism (GMO). There are a total of 20 probes for detecting a GMO in a DNA microarray which can be classified into three categories according to their purpose: the first for screening GMO from un-transgenic plants based on the common elements such as promoter, reporter and terminator genes; the second for specific gene confirmation based on the target gene sequences such as herbicide-resistance or insect-resistance genes; the third for species-specific genes which the sequences are unique for different plant species. To ensure the reliability of this method, different kinds of positive and negative controls were used in DNA microarray. Commercial GM soybean, maize, rapeseed and cotton were identified by means of this method and further confirmed by PCR analysis and sequencing. The results indicate that this method discriminates between the GMOs very quickly and in a cost-saving and more time efficient way. It can detect more than 95% of currently commercial GMO plants and the limits of detection are 0.5% for soybean and 1% for maize. This method is proved to be a new method for routine analysis of GMOs. PMID- 16466913 TI - Efficacy and tolerability of chemotherapy in elderly patients with advanced oesophago-gastric cancer: A pooled analysis of three clinical trials. AB - The aim of this study was to determine the benefits of chemotherapy for oesophago gastric cancer (OGC) in patients 70 years and above (> or =70) in comparison to younger patients. 1080 patients were enrolled into three randomised controlled trials assessing fluorouracil-based combination chemotherapy. Patients received either a platinum-containing regimen (ECF, MCF), PVI 5-FU (protracted venous infusion of 5-fluorouracil)+/-mitomycin C (MMC), or FAMTX. Of the 1080 patients randomised, 257 (23.8%) were aged > or =70 years. There were no significant differences in the incidence of grades 3/4 toxicity between the two cohorts. Objective and symptomatic response rates, failure-free and overall survival were not significantly different. In a multivariate analysis, independent prognostic factors for survival were performance status and locally advanced disease, not age. Patients > or =70 years with OGC obtained similar benefits from palliative chemotherapy with respect to symptomatic response, tumour regression and survival, without increased toxicities. PMID- 16466914 TI - Progestin inhibition of cell death in human breast cancer cell lines. AB - Previously, we have shown that progestins both stimulate proliferation of the progesterone receptor (PR)-rich human breast cancer cell line T47D and protect from cell death, in charcoal-stripped serum-containing medium. To lessen the variability inherent in different preparations of serum, we decided to further characterize progestin inhibition of cell death using serum starvation to kill the cells, and find that progestins protect from serum-starvation-induced apoptosis in T47D cells. This effect exhibits specificity for progestins and is inhibited by the antiprogestin RU486. While progestin inhibits cell death in a dose-responsive manner at physiological concentrations, estradiol-17beta surprisingly does not inhibit cell death at any concentration from 0.001 nM to 1 microM. Progestin inhibition of cell death also occurs in at least two other human breast cancer cell lines, one with an intermediate level of PR, MCF-7 cells, and, surprisingly, one with no detectable level of PR, MDA-MB-231 cells. Further, we have found progestin inhibition of cell death caused by the breast cancer chemotherapeutic agents doxorubicin and 5-fluorouracil. These data are consistent with the building body of evidence that progestins are not the benign hormones for breast cancer they have been so long thought to be, but may be harmful both for undiagnosed cases and those undergoing treatment. PMID- 16466915 TI - Alpha-methylated derivatives of 2-arachidonoyl glycerol: synthesis, CB1 receptor activity, and enzymatic stability. AB - Alpha-methylated analogues of the endogenous cannabinoid, 2-arachidonoyl glycerol (2-AG), were synthesized aiming to the improved enzymatic stability of 2-AG. In addition, the CB1 activity properties of fluoro derivatives of 2-AG were studied. The CB1 receptor activity was determined by the [35S]GTPgammaS binding assay, and the enzymatic stability of alpha-methylated analogues was determined in rat cerebellar membranes. The results indicate that even if the alpha-methylated 2-AG derivatives are slightly weaker CB1 receptor agonists than 2-AG, they are clearly more stable than 2-AG. In addition, the results showed that the replacement of the hydroxyl group(s) of 2-AG by fluorine does not improve the CB1 activity of 2 AG. PMID- 16466916 TI - Synthesis and biological activity of diaryl ether inhibitors of malarial enoyl acyl carrier protein reductase. Part 2: 2'-substituted triclosan derivatives. AB - 2'-Substituted analogs of triclosan have been synthesized to target inhibition of the key malarial enzyme Plasmodium falciparum enoyl acyl carrier protein reductase (PfENR). Many of these compounds exhibit good potency (EC50<500 nM) against in vitro cultures of drug-resistant and drug-sensitive strains of the P. falciparum parasite and modest (IC50=1-20 microM) potency against purified PfENR enzyme. Compared to triclosan, this survey of 2'-substituted derivatives has afforded gains in excess of 20- and 30-fold versus the 3D7 and Dd2 strains of parasite, respectively. PMID- 16466917 TI - Perineural invasion has no prognostic value in patients with invasive breast carcinoma. AB - No studies examining the relationship between perineural invasion (PNI) and clinicopathological factors in invasive breast cancer have been published. Therefore, we investigated this association and analyzed the effect of PNI on disease-free survival (DFS) in patients with invasive breast carcinoma. PNI evaluation was performed on hematoxylin- and eosin-stained tissue sections of surgical specimens from 377 patients. PNI was assessed as positive when cancer cells were seen in the perineurium or neural fascicles. PNI was found in 97 out of 377 patients (25.7%). PNI positivity was 13.9% in T1 tumors and 69.7% in T4 tumors (P<0.001). The incidence of PNI was higher in ductal and mixed type carcinomas than in other histologic types (P=0.013). Vascular invasion, axillary lymph node and progesterone receptor positivity ratios were significantly higher in PNI-positive patients than in PNI-negative ones (P<0.001, P=0.001 and 0.006, respectively). There was no difference between PNI-positive and -negative patients regarding DFS. In conclusion, in invasive breast carcinomas, PNI incidence is lower in small and axilla-negative tumors, and higher in ductal and mixed type and hormone-sensitive tumors; PNI has no prognostic importance. PMID- 16466918 TI - A Wnt canon orchestrating osteoblastogenesis. AB - Several transcription factors have been identified that control the differentiation of osteoblasts; however, relatively little is known about the signaling pathways involved in regulating the differentiation process. Recently, the canonical Wnt-beta-catenin pathway has been implicated in osteoblastogenesis. This review focuses on the role of the canonical Wnt-beta-catenin pathway during embryonic development, where it is required for the differentiation of osteoblasts from a precursor that is shared with the chondrocyte lineage and the requirement of this pathway during postnatal life in bone homeostasis. The recent findings covered in this review are major advances in our understanding of skeletal development and promise new therapeutic avenues for tissue engineering and treatment of osteoporosis. PMID- 16466919 TI - Revision total knee arthroplasty using the Kinemax Plus Superstabilizer prosthesis: minimum 10-year follow-up. AB - The results of revision total knee arthroplasty using the Kinemax Plus Superstabilizer prosthesis at a minimum of 10 years are reviewed. The implant was designed specifically for revision surgery with integral distal and posterior femoral build-ups and a modular metal reinforcement rod within the tibial polyethylene post. Eighteen consecutive knees revised for aseptic loosening or instability were studied prospectively and reviewed at 10-13 years (mean 11.6 years). Mean HSS scores improved from 51 to 86, KS knee scores from 40 to 83 and KS function scores from 48 to 82, with 83% good and excellent results. There was one re-revision for recurrent instability. Additional distal femoral add-ons were needed in 33% and posterior add-ons were needed in 22% of cases. No post breakage or deformation occurred. The design rationale has proven successful with satisfactory clinical and radiographic outcomes. PMID- 16466920 TI - New derivatives of silybin and 2,3-dehydrosilybin and their cytotoxic and P glycoprotein modulatory activity. AB - Large series of O-alkyl derivatives (methyl and benzyl) of silybin and 2,3 dehydrosilybin was prepared. Selective alkylation of the silybin molecule was systematically investigated. For the first time we present here, for example, preparation of 19-nor-2,3-dehydrosilybin. All prepared silybin/2,3-dehydrosilybin derivatives were tested for cytotoxicity on a panel of drugs sensitive against multidrug resistant cell lines and the ability to inhibit P-glycoprotein mediated efflux activity. We have identified effective and relatively non-cytotoxic inhibitors of P-gp derived from 2,3-dehydrosilybin. Some of them were more effective inhibitors at concentrations lower than a standard P-gp efflux inhibitor cyclosporin A. Another group of 2,3-dehydrosilybin derivatives also had better inhibitory effects on P-gp efflux but a cytotoxicity comparable with that of parent 2,3-dehydrosilybin. Structural requirements for improving inhibitory activity and reducing toxicity of 2,3-dehydrosilybin were established. Effect of E-ring substitution as well as an influence of the substituent size at the C-7-OH position of A-ring on P-gp-inhibitory activity was evaluated for the first time in this study. PMID- 16466921 TI - Ether derivatives of 3-piperidinopropan-1-ol as non-imidazole histamine H3 receptor antagonists. AB - A series of aliphatic and aromatic ether derivatives of 3-piperidinopropan-1-ol has been prepared by four different methods. The ethers obtained were evaluated for their affinities at recombinant human histamine H3 receptor, stably expressed in CHO-K1 or HEK 293 cells. All compounds investigated show from moderate to high in vitro affinities in the nanomolar concentration range. Selected compounds were investigated under in vivo conditions after oral administration to mice. Some proved to be highly potent and orally available histamine H3 receptor antagonists. The most potent antagonists in this series have been in vitro the 4 (1,1-dimethylpropyl)phenyl ether 19 (hH3R K(i) = 8.4 nM) and in vivo the simple ethyl ether 2 (ED50 = 1.0mg/kg). PMID- 16466922 TI - Synthesis and structure-activity relationships (SARs) of 1,5-diarylpyrazole cannabinoid type-1 (CB(1)) receptor ligands for potential use in molecular imaging. AB - Cannabinoid type-1 (CB(1)) receptor ligands, derived from the 1,5-diarylpyrazole core template of rimonabant (Acomplia), have been the focus of several studies aimed at examining structure-activity relationships (SARs). The purpose of this study was to design and synthesize a set of compounds based on the 1,5 diarylpyrazole template while focusing on the potential for discovery of CB(1) receptor radioligands that might be used as probes with in vivo molecular imaging. Each synthesized ligand was evaluated for potency as an antagonist at CB(1) and cannabinoid type-2 (CB(2)) receptors in vitro using a GTPgamma(35)S binding assay. clog P values were calculated with Pallas 3.0. The antagonist binding affinities (K(B)) at CB(1) receptors ranged from 11 to >16,000 nM, CB(1) versus CB(2) selectivities from 0.6 to 773, and clog Ps from 3.61 to 6.25. An interesting new ligand, namely N-(piperidin-1-yl)-1-(2-bromophenyl)-5-(4 methoxyphenyl)-4-methyl-1H-pyrazole-3-carboxamide (9j), emerged from the synthesized set with appealing properties (K(B)=11 nM; CB(1) selectivity>773; clog P=5.85), for labeling with carbon-11 and development as a radioligand for imaging brain CB(1) receptors in vivo with positron emission tomography (PET). PMID- 16466923 TI - Design, synthesis, and evaluation of new type of L-amino acids containing pyridine moiety as nitric oxide synthase inhibitor. AB - New amino acids 7-12 were designed and synthesized as candidate inhibitors of human nitric oxide synthase (NOS). The 2-aminopyridine-containing l-amino acids 8 had potent inhibitory activity toward all of the human NOS isozymes. However, the regioisomers 9 and 10, and 2-methylpyridine-containing compound 11 had much lower inhibitory activity. Human NOS isozymes were also inhibited by 7, which lacks an amino group on the pyridine moiety. A computational docking study was carried out to investigate the mechanism of the inhibitory effect. PMID- 16466924 TI - A novel kind of nitrogen heterocycle compound induces apoptosis of human chronic myelogenous leukemia K562 cells. AB - The effects of a novel kind of nitrogen heterocycle compound, which was synthesized in our laboratory previously, on human chronic myelogenous leukemia K562 cells were investigated. The morphological changes were observed by Acridine orange (AO) staining. The screened results through DNA fragmentation and the Annexin V-FITC/PI staining assay showed that compound 8 blocked cell cycles at G(1) phase which led to apoptosis. The increase of caspase-3, 8, and 9 was detected, indicating that both of death-receptor and mitochondria-pathways were activated. Compound 8 induced a biphasic alteration in mitochondrial membrane potential of K562 cells. A dramatic elevation of Ca(2+) was also observed. In addition, a transient increase of ROS was also involved in the process. This study showed that compound 8 might be a potential chemopreventive agent for chronic myelogenous leukemia. It would guide our future work to synthesize more compounds derived from compound 8, which might have better effect, and to determine the target protein. Moreover, it might also provide a background mechanism for the introduction of this new type of promising therapeutic agent. PMID- 16466925 TI - Determination of the oxidation state for iron oxide minerals by energy-filtering TEM. AB - The oxidation state of iron oxide nanoparticles was determined using the two principally different technical realisations of energy filtering TEM, in one case using the JEOL 3010 equipped with a LaB6 cathode and a post-column GIF and in the second, the newly designed LIBRA 200FE equipped with an corrected in-column 90 degrees energy filter and a field emission gun (Schottky emitter). The samples studied were oxide-coated iron nanoparticles, and iron oxide inclusions in feldspars in granites. Five possible candidates exist for the iron-oxide phases: FeO, alpha-Fe2O3 (hematite), gamma-Fe2O3 (maghemite), Fe3O4 (magnetite) or alpha FeO(OH) (goethite). Fingerprinting the O K-edge ELNES allows to distinguish between oxide phases with the same stochiometry and enables to make a first selection of possible candidates. The additional determination of the chemical composition allows unique identification of the phase present. For the oxide coated iron nanoparticles the most probable iron oxide phase of the shell is maghemite, which was additionally confirmed by HRTEM studies. The second studied system were iron oxide needles in alkali feldspar, where we obtained hematite as the most probable phase. There we additionally demonstrated the drastic changes of the ELNES of the O K-edge for the alkali feldspar and iron oxide needle by spatially resolved EELS. PMID- 16466926 TI - Electron beam damage studies of synthetic 6-line ferrihydrite and ferritin molecule cores within a human liver biopsy. AB - In order to achieve an accurate understanding of the crystal structure of 6-line ferrihydrite (6LFh) and ferritin molecule cores within a human liver biopsy using transmission electron microscopy (TEM), electron beam damage should be considered. For the case of 6LFh, the electron energy loss near-edge structure (ELNES) of core ionisation edges in the electron energy loss spectrum (EELS) combined with multiple linear least-square (MLLS) fitting of reference spectra together with analysis of selected area electron diffraction (SAED) patterns suggests that the iron in 6LFh is solely octahedrally coordinated Fe3+. With increasing electron dose, an increasing percentage of this octahedrally coordinated Fe3+ migrates to tetrahedral sites. When the dose exceeds 3 x 10(8) electrons/nm2, Fe2+ is found to be present in the material. This method also indicates that the iron in ferritin molecule cores within a human liver biopsy is the same as in 6LFh, entirely Fe3+ in octahedral coordination with oxygen. Again the percentage of octahedrally coordinated Fe3+ decreases as the accumulated electron dose increases and Fe2+ is produced in the liver biopsies when the electron dose exceeds 10(6)electrons/nm2. PMID- 16466927 TI - Rapana thomasiana hemocyanin (RtH): comparison of the two isoforms, RtH1 and RtH2, at 19A and 16A resolution. AB - Three-dimensional (3D) reconstructions of the two 8.4 MDa Rapana thomasiana hemocyanin isoforms, RtH1 and RtH2, have been obtained by cryoelectron microscopy of molecules embedded in vitreous ice and single particle image processing. The final 3D structures of the RtH1 and RtH2 didecamers at 19 A and 16 A resolution, respectively, are very similar to earlier reconstructions of gastropodan hemocyanins, revealing structural features such as the obliquely oriented subunits, the five- and two-fold symmetrical axes. Three new interactions are defined; two of them connecting the arch and the wall while the third is formed between the collar and the wall. The collar-wall connection and one of the arch wall connections are positioned between two individual subunit dimers, while the second arch-wall connection is located between two subunits within the subunit dimer. All three interactions establish connections to the first tier of the wall. Furthermore, for each interaction we have allocated two first tier functional units most likely involved in forming the connections. PMID- 16466928 TI - HRTEM and EELS study of Y2O3/MgO thin films. AB - Y2O3 thin films deposited on (001)-MgO substrate have been investigated by high resolution transmission electron microscopy (HRTEM) and electron energy loss spectroscopy. Digital processing of the HRTEM images reveals the presence of grains with a crystallographic structure different from that of the rest of the film (Ia3). The spectrum imaging technique has been applied in vicinity of the Y2O3/MgO interface to get a better knowledge of the phases nucleated on the substrate surface. Fine structures of the O K-edge have been studied in detail; actually two kinds of spectra have been detected in the yttrium oxide thin film. These spectra have been compared to self-consistent full multiple scattering calculations (SC-FMS). One family of spectra has then been associated to the well known Ia3 structure. The other family of spectra has been compared to calculations performed for the other known structures (such as hexagonal or monoclinic) of Y2O3 with a little success. We have finally compared these spectra to calculations performed with a particular atomic arrangement (octahedral) of Y and O atoms, which leads to a good match between experimental and calculated spectra. Our results emphasize the benefit of coupling several techniques such as HRTEM, EELS and SC-FMS for the determination of structures at the nanometric scale. PMID- 16466929 TI - Redox proteomics identification of oxidatively modified hippocampal proteins in mild cognitive impairment: insights into the development of Alzheimer's disease. AB - Mild cognitive impairment (MCI) is generally referred to the transitional zone between normal cognitive function and early dementia or clinically probable Alzheimer's disease (AD). Oxidative stress plays a significant role in AD and is increased in the superior/middle temporal gyri of MCI subjects. Because AD involves hippocampal-resident memory dysfunction, we determined protein oxidation and identified the oxidized proteins in the hippocampi of MCI subjects. We found that protein oxidation is significantly increased in the hippocampi of MCI subjects when compared to age- and sex-matched controls. By using redox proteomics, we determined the oxidatively modified proteins in MCI hippocampus to be alpha-enolase (ENO1), glutamine synthetase (GLUL), pyruvate kinase M2 (PKM2) and peptidyl-prolyl cis/trans isomerase 1 (PIN1). The interacteome of these proteins revealed that these proteins functionally interact with SRC, hypoxia inducible factor 1, plasminogen (PLG), MYC, tissue plasminogen activator (PLAT) and BCL2L1. Moreover, the interacteome indicates the functional involvement of energy metabolism, synaptic plasticity and mitogenesis/proliferation. Therefore, oxidative inactivation of ENO1, GLUL and PIN1 may alter these cellular processes and lead to the development of AD from MCI. We conclude that protein oxidation plays a significant role in the development of AD from MCI and that the oxidative inactivation of ENO1, GLUL, PKM2 and PIN1 is involved in the progression of AD from MCI. The current study provides a framework for future studies on the development of AD from MCI relevant to oxidative stress. PMID- 16466930 TI - UVA1 radiation (340-400 nm) interferes with the perforin-granule system of CD8hi+ cytotoxic T lymphocytes in vitro. AB - UVA1-irradiation was introduced as an innovative and effective phototherapy of atopic dermatitis (AD) and other skin diseases. In AD, a defect of a central apoptosis inducing effector system involved in immunoregulation and immune defense, i.e., the system of perforin-granules in cytotoxic T lymphocytes (CTL), was recently reported: perforin-reduction and perforin-hyperreleasability. We now investigated UVA1-effects on the perforin-granule system in vitro. Peripheral blood CTLs were exposed in vitro to 10-100 J/cm2 UVA1 (340-400 nm), and to 30-150 mJ/cm2 UVB (280-315 nm) as a control. A time-dependent perforin-granule release was induced by phorbol 12-myristate 13-acetate (PMA) and ionomycin. This release was inhibited dose-dependently by UVA1, but not by UVB. An UVA1-dose dependent pattern of sensitive (80%) and insensitive (20%) individuals was found. The kinetics of perforin release in AD-CTLs, i.e. hyperreleasability, was normalized by 50 J/cm2 UVA1 in vitro. Sodium azide as a quencher of reactive oxygen species prevented the UVA1-mediated inhibition of perforin-granule release. Our data demonstrate for the first time a dose- and wavelength-dependent UVA1-effect in vitro on a major effector system of cytotoxic lymphocytes, the system of perforin granules. This might contribute to the further understanding of immunomodulatory UVA1-effects in vivo. PMID- 16466931 TI - Using PET H2O15 brain imaging to study the functional-anatomical correlates of non-human primate communication. AB - In this article, we describe methods for using oxygen-15 water (H2O15) positron emission tomography (PET) to explore the functional neuroanatomy of cognition in awake, non-human primates. The discussion is based on a recent study designed to identify regions in the monkey brain associated with perceiving auditory stimuli, and species-specific calls, in particular [Gil-da-Costa et al., Proc. Natl. Acad. Sci. USA 101 (2004) 17516-17521]. Details are provided concerning critical aspects of the experimental paradigm, including pre-scanning habituation sessions to acclimate the animals to the PET scanner environment, and details of a pilot study to determine the auditory stimulus parameters necessary to produce robust activity in brain regions known to process auditory information (belt and parabelt regions of monkey auditory cortex). Methods for acquiring and analyzing PET data to identify significant regions of brain activity in single animals are also presented. PMID- 16466932 TI - Pianists duet better when they play with themselves: on the possible role of action simulation in synchronization. AB - Ensemble musicians play in synchrony despite expressively motivated irregularities in timing. We hypothesized that synchrony is achieved by each performer internally simulating the concurrent actions of other ensemble members, relying initially on how they would perform in their stead. Hence, musicians should be better at synchronizing with recordings of their own earlier performances than with others' recordings. We required pianists to record one part from each of several piano duets, and later to play the complementary part in synchrony with their own or others' recordings. The pianists were also asked to identify their own recordings. The pianists were better at synchronizing with their own than with others' performances, and they were able to recognize their own recordings. Furthermore, synchronization accuracy and recognition were correlated: Pianists who were relatively accurate at synchronizing with their own performances were also good at recognizing them. Thus, action simulation may underlie both synchronization and self-recognition. PMID- 16466933 TI - Cortical reorganization during PASAT task in MS patients with preserved working memory functions. AB - Cortical reorganization in multiple sclerosis (MS) is defined as a compensatory mechanism which requires MS patients to overactivate specific brain areas in order to perform the task as controls. To investigate this process with the Paced Auditory Serial Addition Test (PASAT) task, we selected 15 MS patients who performed the PASAT task within-normal limits and 10 healthy controls. Once selected, we used functional magnetic resonance imaging (fMRI) to investigate brain areas involved in PASAT performance in both groups. Results showed that the task activated the left frontal (BA6 and 9) and parietal cortex (BA7 and 40) in both groups, but MS patients showed a stronger activation in the left prefrontal cortex (BA9, 44 and 45) when compared with controls. These results confirmed those obtained post hoc by Audoin et al. [Audoin, B., Ibarrola, D., Ranjeva, J.P., Confort-Gouny, S., Malikova, I., Ali-Cherif, A.M., Pelletier, J., Cozzone, P., et al., 2003. Compensatory cortical activation observed by fMRI during cognitive task at the earliest stage of MS. Hum. Brain Mapp. 20, 51-58], and we interpreted this as showing true cortical reorganization. PMID- 16466934 TI - Effect of repetitive transcranial magnetic stimulation applied over the premotor cortex on somatosensory-evoked potentials and regional cerebral blood flow. AB - Somatosensory-evoked potentials (SEPs) are attenuated by movement. This phenomenon of 'gating' reflects sensorimotor integration for motor control. The frontal N30 component after median nerve stimulation was shown to be reduced in amplitude prior to hand movement. To investigate the mechanism of this sensory gating, we recorded median SEPs immediately before and after application of monophasic very low-frequency (0.2 Hz) repetitive transcranial magnetic stimulation (rTMS) of 250 stimuli over motor cortex (MC), premotor cortex (PMC), or supplementary motor area (SMA) in 9 healthy volunteers. The stimulus intensity for MC or PMC was set 85% of the resting motor threshold for the hand muscle, and that for SMA was at the active motor threshold for the leg muscle. SEPs showed significant increases in amplitudes of the frontal N30 component after PMC stimulation, but not after SMA or MC stimulation. Low-frequency (1 Hz) biphasic stimulation over PMC showed no significant N30 changes in 6 out of 9 subjects tested, indicating the effect being specific for 0.2 Hz monophasic stimulation. To examine the functional anatomy of the N30 change, single photon emission computed tomography was performed immediately before and after monophasic 0.2 Hz rTMS over PMC in all the 9 subjects. Regional cerebral blood flow showed significant increases mainly in PMC and prefrontal cortex, indicating the involvement of these cortical areas in sensory input gating for motor control. PMID- 16466935 TI - BOLD responses to stimuli: dependence on frequency, stimulus form, amplitude, and repetition rate. AB - A quantitative theory is developed for the relationship between stimulus and the resulting blood oxygen level-dependent (BOLD) functional MRI signal. The relationship of stimuli to neuronal activity during evoked responses is inferred from recent physiology-based quantitative modeling of evoked response potentials (ERPs). A hemodynamic model is then used to calculate the BOLD response to neuronal activity having the form of an impulse, a sinusoid, or an ERP-like damped sinusoid. Using the resulting equations, the BOLD response is analyzed for different forms, frequencies, and amplitudes of stimuli, in contrast with previous research, which has mostly concentrated on sustained stimuli. The BOLD frequency response is found to be closely linear in the parameter ranges of interest, with the form of a low-pass filter with a weak resonance at approximately 0.07 Hz. An improved BOLD impulse response is systematically obtained which includes initial dip and post-stimulus undershoot for some parameter ranges. It is found that the BOLD response depends strongly on the precise temporal course of the evoked neuronal activity, not just its peak value or typical amplitude. Indeed, for short stimuli, the linear BOLD response is closely proportional to the time-integrated activity change evoked by the stimulus, regardless of amplitude. It is concluded that there can be widely differing proportionalities between BOLD and peak activity, that this is the likely reason for the low level of correspondence seen experimentally between ERP sources and BOLD measurements and that non-BOLD measurements, such as ERPs, can be used to correct for this effect to obtain improved activity estimates. Finally, stimulus sequences that optimize the signal-to-noise ratio in event related BOLD fMRI (efMRI) experiments are derived using the hemodynamic transfer function. PMID- 16466936 TI - Network modulation by the subthalamic nucleus in the treatment of Parkinson's disease. AB - Deep brain stimulation of the subthalamic nucleus (STN DBS) has become an accepted tool for the treatment of Parkinson's disease (PD). Although the precise mechanism of action of this intervention is unknown, its effectiveness has been attributed to the modulation of pathological network activity. We examined this notion using positron emission tomography (PET) to quantify stimulation-induced changes in the expression of a PD-related covariance pattern (PDRP) of regional metabolism. These metabolic changes were also compared with those observed in a similar cohort of patients undergoing STN lesioning. We found that PDRP activity declined significantly (P < 0.02) with STN stimulation. The degree of network modulation with DBS did not differ from that measured following lesioning (P = 0.58). Statistical parametric mapping (SPM) revealed that metabolic reductions in the internal globus pallidus (GPi) and caudal midbrain were common to both STN interventions (P < 0.01), although declines in GPi were more pronounced with lesion. By contrast, elevations in posterior parietal metabolism were common to the two procedures, albeit more pronounced with stimulation. These findings indicate that suppression of abnormal network activity is a feature of both STN stimulation and lesioning. Nonetheless, these two interventions may differ metabolically at a regional level. PMID- 16466938 TI - A comparison between the short term and long term benefits of screening for abdominal aortic aneurysms from the Huntingdon Aneurysm screening programme. AB - BACKGROUND: The UK Multicentre Aneurysm Screening Study (MASS) showed a 44% reduction in AAA-related mortality after 4 years and predicted an increased number of deaths prevented in the longer term. We aim to compare the 5 and 13 years benefit from aneurysm screening in the Huntingdon Aneurysm screening programme. METHODS: Incidence and mortality of ruptured AAA (RAAA) after 5 and 13 years of screening in a population based aneurysm screening program. RESULTS: Five years of screening resulted in a reduction in the incidence of RAAA of 49% (95% CI: 3-74%). Nine out of 11 ruptures in the invited group did not survive (mortality 82%; 95% CI: 48-98%) compared to 38 non-survivors from 51 ruptures in the control group (mortality 75%; 95% CI: 60-86%). Five years of screening resulted in an RAAA-related mortality reduction of 45% (95% CI: -15 to 74%). After 13 years of screening the incidence of RAAA was reduced by 73% (95% CI: 58 82%). Twenty-one out of 29 ruptures in the invited group did not survive (mortality 72%; 95% CI: 53-87%) compared to 64 non-survivors from 82 ruptures in the control group (mortality 78%; 95% CI: 68-86%). Thirteen years screening resulted in a reduction of mortality from RAAA of 75% (95% CI: 58-84%). The number needed to screen to prevent one death reduced from 1380 after 5 years to 505 after 13 years. The number of elective AAA operations needed to prevent one death reduced from 6 after 5 years to 4 after 13 years. CONCLUSION: AAA screening becomes increasingly beneficial as screening continues over the longer term. Benefits continue to increase after screening has ceased. PMID- 16466937 TI - Rapid induction of specific associative behavioral memory by stimulation of the nucleus basalis in the rat. AB - Hypothesized circuitry enabling behavioral memory formation can be tested by its direct activation in the absence of normal experience. Neuromodulation via the cortical release of acetylcholine by the nucleus basalis (NB) is hypothesized to be sufficient to induce specific, associative behavioral memory. Previously, we found that tone paired with stimulation of the nucleus basalis (NBs) for 3000 trials over 15 days induced such memory, supporting the hypothesis. However, as standard associative memory can be established much more rapidly, we asked whether NB-induced memory develops rapidly. Adult male Sprague-Dawley rats, trained and tested in the same calm, waking state, were divided into Paired (n=5) and control (n=4) groups, each of which received a single session of 200 trials of an 8.0 kHz conditioned stimulus (CS) either paired with NBs or with unpaired presentation of NBs. Respiration, cardiac activity, and evoked potentials in the primary auditory cortex (ACx) were recorded. Memory and its degree of specificity were assessed 24 h later by presenting tones of various frequencies (1-15 kHz) in the absence of NBs to yield behavioral frequency generalization gradients. Behavioral responses to test tones consisted of interruption of ongoing respiration and changes in heart rate. Post-training behavioral generalization gradients exhibited response peaks centered on the CS frequency for the Paired group alone. Tone evoked potentials from the ACx also developed CS-specific plasticity. The findings indicate that NB induction of specific behavioral associative memory, like normal memory, can develop rapidly and is accompanied by specific cortical plasticity, supporting the view that NB engagement during normal learning produces memory. PMID- 16466939 TI - Cognitive function remains unchanged after endarterectomy of unilateral internal carotid artery stenosis under local anaesthesia. AB - OBJECTIVE: To assess changes in cognitive function and affective state following carotid endarterectomy (CEA) for high-degree unilateral internal carotid artery stenosis. METHODS: In 33 patients, a CEA was performed under local anaesthesia for a high-grade unilateral stenosis of the internal carotid artery (group A). Twenty-five patients underwent surgery for peripheral arterial occlusive disease under regional anaesthesia served as controls (group B). Patients with neurological deficits due to previous strokes or dementia were excluded. Intelligence level was assessed preoperatively. Cognitive tests were applied preoperatively and postoperatively (3-5 days after surgery) and after 4 months follow-up. Confounding factors, including anxiety and depression, were checked through questionnaires and interviews. RESULTS: No perioperative neurological complication occurred following CEA. Patients in group A showed a significant postoperative deterioration only in one sub-test. There was no significant change in anxiety and depression during follow up. The control group B had no significant changes in cognitive test performance. Anxiety improved significantly postoperatively, but increased again at the end of the study. There was no significant difference between the groups over time. CONCLUSION: Cognitive function does not change following CEA of a unilateral internal carotid stenosis. PMID- 16466940 TI - Nanocomposite containing bioactive peptides promote endothelialisation by circulating progenitor cells: an in vitro evaluation. AB - OBJECTIVE: The formation of an endothelial cell layer on the luminal surface of cardiovascular devices, especially bypass grafts, is an important attribute in order to improve their patency. Endothelial progenitor cells (EPCs) have a potential role in the endothelialisation of bypass grafts. We hypothesised that a novel approach to improve endothelialisation of bypass grafts by EPCs would be the creation on the graft lumen of a microenvironment that supports EPC adhesion and differentiation. METHODS: A new generation of nanocomposite based on silsesquioxane in the form of polyhedral oligomeric silsesquioxane (POSS) nanocages which incorporate bioactive peptides (RGD) was made into sheets. Peripheral blood mononuclear cells (PBMCs) containing EPCs isolated from six consenting young, healthy, adult volunteers were then plated both on (1) sheets of the nanocomposite with the bioactive peptide, (2) sheets of the nanocomposite without the bioactive peptide, (3) culture dishes as control and then cultured in presence of vascular endothelial growth factor (VEGF). Confirmation of endothelial and EPCs markers was carried out using fluorescence-activated cell sorter (FACS) analysis, reverse transcription polymerase chain reaction (RT-PCR) and immunostaining. RESULTS: One to two percent of PBMCs expressed CD34 as determined by FACS analysis. Cells were demonstrated to express mRNA for the EPC markers CD34, platelet-endothelial cell adhesion molecule-1 (CD31), CD133 and vascular endothelial growth factor receptor-2(FlK-1/KDR). Endothelial cell-colony forming units were formed between day 5 and day 7 after plating. Colonies were confirmed to be endothelial like cells by immunostaining. There were significantly greater numbers of EPC colonies on the bioactive nanocomposites as compared to the nanocomposite alone and the uncoated dishes. CONCLUSION: We report a new nanocomposite based biomaterial that has been demonstrated, in vitro, to promote endothelialisation from PBMCs containing EPCs. PMID- 16466941 TI - Femoral pseudoaneurysm in drug addicts--excision without revascularization is a viable option. AB - PURPOSE: To present a series of patients presenting with femoral pseudoaneurysm. RESULTS: Seventeen patients who presented with a femoral pseudoaneurysm during a 1 year period were included in this study. Parenteral drug abuse was the most common aetiological factor. The femoral artery was most commonly involved at its bifurcation. Sixteen patients (94%) had excision of the pseudoaneurysm with ligation of vessel and debridement without any revascularization and one patient (6%) had reverse saphenous grafting after excision and ligation of vessels. Four amputations (23%) were performed. Three (17%) were major limb amputations, which included one above knee and two below knee amputations. Four patients (23%) developed intermittent claudication. CONCLUSION: Excision of the pseudoaneurysm with ligation of vessels and wide debridement without immediate revascularization in infected pseudoaneurysms is a safe and effective treatment. PMID- 16466942 TI - Does the type, number or combinations of traditional cardiovascular risk factors affect early outcome after carotid endarterectomy? AB - PURPOSE: The present study was undertaken in order to assess the 30-day complication rate of carotid endarterectomy (CEA) in relation to the patients' cardiovascular risk factors. METHODS: Cardiovascular risk factors, operative details, morbidity and mortality of 1002 carotid endarterectomies in 852 patients were prospectively recorded in a database. The indications for surgery were asymptomatic >or=75% or symptomatic >or=50% internal carotid stenosis when other causes of stroke were excluded. Exclusion criteria were intervention for post-CEA restenosis, post-irradiation lesions, kinking of the internal carotid artery, external carotid artery stenosis, endovascular and simultaneous cardiac procedures. RESULTS: The 30-day combined minor and major stroke and death rate was 2.7% (27/1002). Significant risk factors in logistic regression model were diabetes (stroke and death rate=5.7%, p=0.002, OR=3.31), the simultaneous presence of three cardiovascular risk factors (stroke and death rate=5.3%, p=0.012, OR=3.11) and the combination diabetes, hypertension and hyperlipidemia (stroke and death rate=9.4%, p=0.001, OR=4.22). CONCLUSIONS: Traditional cardiovascular risk factors significantly affect the 30-day stroke and death rate after carotid endarterectomy. PMID- 16466943 TI - Primary leiomyosarcoma of the great saphenous vein: case report. AB - OBJECTIVE: To describe a case of primary leiomyosarcoma of the great saphenous vein. PATIENT RECORD: A 59-year-old Chinese lady presented with two painful lumps in the right thigh in the line of the great saphenous vein. At surgery, in September 2004, two tumors in the right great saphenous vein above the knee were excised with the intervening normal vein. Histopathological studies confirmed both masses as leiomyosarcoma. Radiotherapy was given postoperatively. The patient has been followed-up for 300 days after surgery with no evidence of local or distant metastasis. CONCLUSION: Leiomyosarcoma of the great saphenous vein can exist at more than one site. Tumor resection and radiotherapy was associated with good patient prognosis. PMID- 16466945 TI - Unique genetic profile of hereditary haemochromatosis in Russians: High frequency of C282Y mutation in population, but not in patients. PMID- 16466946 TI - The intrinsic pathway of coagulation is essential for thrombus stability in mice. AB - Blood coagulation is a highly regulated process involving interactions between platelets, plasma coagulation factors, and the vessel wall. During coagulation in vivo, fibrin formation is thought to be initiated when plasma factor VIIa forms a complex with the membrane protein tissue factor. Coagulation factor XII (FXII, Hageman factor) is required for some in vitro coagulation systems; however, FXII deficiency is not associated with hemorrhage, leading to the conclusion that it is not necessary for hemostasis. We generated FXII-deficient mice to study the contributions of FXII to thrombosis and hemostasis in arterial injury models and in models of acute arterial occlusion. FXII-deficient mice do not experience excessive injury-related bleeding; however, intravital fluorescence microscopy and blood flow measurements in three separate arterial beds revealed a severe defect in formation and stabilization of platelet-rich occlusive thrombi induced by different methods of injuries. Similar findings were observed for mice deficient in factor XI, a substrate of activated FXII. Infusion of human FXII into FXII null mice restored thrombus formation. These findings demonstrate that FXII-mediated fibrin formation is crucial for pathological arterial thrombosis but not for hemostasis and suggest that FXII could be an ideal target for safe anticoagulation. PMID- 16466947 TI - Novel and Mediterranean beta thalassemia mutations in the indigenous Northern Ireland population. AB - beta thalassemia is one of the most common genetic diseases worldwide resulting from aberrant beta-globin chain production. It is highly prevalent in regions with endemic malaria, but it is also present at low frequency in the indigenous populations of non-tropical areas such as Britain. Screening beta thalassemia trait individuals from Northern Ireland has detected 2 Mediterranean mutations, 39 (C --> T) and IVS-I-110 (G --> A); the previously reported IVS-II-850 (G --> A) mutation originally described in individuals of Scottish/English ancestry; and 2 novel mutations, initiation codon A --> C and 109 delG. Haplotype analysis indicates that the Mediterranean mutations are present on previously described haplotypes, suggesting that they have arisen due to migration. It remains to be established whether the novel mutations have arisen de novo in Northern Ireland. PMID- 16466948 TI - The platelet P2 receptors in arterial thrombosis. AB - ADP and ATP play a crucial role in hemostasis and thrombosis and their receptors are potential targets for antithrombotic drugs. The ATP-gated channel P2X1 and the two G protein-coupled P2Y1 and P2Y12 ADP receptors selectively contribute to platelet aggregation. Due to its central role in the formation and stabilization of a thrombus, the P2Y12 receptor is a well established target of antithrombotic drugs like clopidogrel which has proved efficacious in many clinical trials and experimental models of thrombosis. Competitive P2Y12 antagonists have also been shown to be effective in experimental thrombosis as well as in several clinical trials. Studies in P2Y1 and P2X1 knock-out mice and experimental thrombosis models using selective P2Y1 and P2X1 antagonists have shown that, depending on the conditions, these receptors could also be potential targets for new antithrombotic drugs. Since both P2X1 and P2Y1 receptor inhibition result in milder prolongation of the bleeding time as compared to P2Y12 inhibition, the idea is put forward that combination of P2 receptor antagonists could improve efficacy with diminished hemorrhagic risk. However, further studies are required to validate such a point of view. PMID- 16466949 TI - Formation and fate of platelet microparticles. AB - There is increasing evidence that platelet microparticles participate in thrombus formation. Yet the origin of platelet microparticles and their fate in the circulation remain poorly defined. It is unknown, for example, whether circulating platelet microparticles found in healthy individuals are derived from activated platelets or generated during megakaryopoiesis. The life span of platelet microparticles and the mechanism of their clearance have also not been determined. This article addresses these fundamental aspects of the physiology of platelet microparticles. PMID- 16466950 TI - Prevalence of -alpha(3.7) and alpha alpha alpha(anti3.7) alleles in sickle cell trait and beta-thalassemia patients in Mexico. AB - The aim of this study was to determine the frequency of alpha-globin gene mutations in three groups of Mexican unrelated individuals. The first two groups were normal and sickle cell trait individuals from the Costa Chica region, a place with a 12.8% frequency of HbS carriers, and the third group comprised of Mexican mestizo patients with beta-thalassemia. We searched for -alpha(3.7) and alpha(4.2) alpha(+)-thalassemia deletion alleles, as well as the alpha alpha alpha(anti3.7) triplication through long-gap PCR. The alleles -alpha(3.7) and alpha alpha alpha(anti3.7) were found in the heterozygote state only; 19% of the normal subjects had the -alpha(3.7) allele, and 2% showed the alpha alpha alpha(anti3.7) allele. In individuals with the sickle cell trait, 17% had the alpha(3.7) deletion, and the alpha alpha alpha(anti3.7) triplication was observed in 3% of these individuals. We revealed that 16% of the subjects with beta thalassemia showed the -alpha(3.7) deletion and 28% the alpha alpha alpha(anti3.7) triplication. The -alpha(4.2) deletion was not detected in any individual. The frequency of the -alpha(3.7) allele was roughly the same in the three groups studied; this can be explained by the fact that the three groups have common genes from Africa and the Mediterranean, where a high prevalence of alpha(+)-thalassemia has been observed. To our knowledge, the frequency of alpha alpha alpha(anti3.7) triplication observed in the Mexican beta-thalassemia patients is the highest reported. As the -alpha(3.7) and alpha alpha alpha(anti3.7) alleles are very common in our selected populations, we believe that there is a need to investigate systematically the alpha-globin gene mutations in all hemoglobinopathies in the Mexican population. PMID- 16466951 TI - Role of tissue factor in hemostasis and thrombosis. AB - Tissue factor (TF) is a transmembrane glycoprotein that functions as the primary cellular initiator of blood coagulation. Perivascular cells express TF and provide a hemostatic barrier to limit hemorrhage after vessel injury. In addition, TF is expressed in a tissue-specific manner with high levels in vital organs, such as the heart and lung. TF expression in these tissues may provide additional hemostatic protection from mechanical injury to blood vessels. Recent studies have also detected TF in the blood. This circulating TF is present in the form of microparticles (MPs), which are membrane vesicles shed from cells, and possibly platelets. At present, the cell types that contribute to this pool of TF positive MPs have not been fully defined. Monocytes, endothelial cells and platelets are the most likely sources of this circulating TF. However, TF positive MPs represent only a minor subset of circulating MPs. Importantly, TF negative MPs also possess procoagulant activity. In various diseases, such as sepsis and cancer, TF is expressed by vascular cells and this leads to thrombosis. Levels of circulating TF are also elevated in these diseases and may contribute to thrombosis. Recent studies have analyzed the role of TF-positive MPs in thrombus propagation using different in vivo models. Circulating TF was found to contribute to thrombosis in some models but not others. Inhibition of TF activity in patients with TF expression in vascular cells and with elevated levels of circulating TF may decrease thrombosis associated with a variety of diseases. PMID- 16466952 TI - The complex between activated protein C and protein C inhibitor: A clinically useful indicator of aortic aneurysms? AB - The concentration of the complex between activated protein C and the protein C inhibitor reflects the degree of activation of blood coagulation. A sandwich method has been devised that measures the complex concentration in blood plasma. A key feature of the method is that the catching monoclonal antibody recognizes a complex-dependent neoepitope in PCI, which is a prerequisite, since the concentration of uncomplexed PCI is approximately 10(4)-fold higher than that of the complex. In patients with atherosclerotic disease, those with aortic aneurysms exhibit a three-fold increase in complex concentration compared to that of normal subjects. PMID- 16466953 TI - ThPTR2, a di/tri-peptide transporter gene from Trichoderma harzianum. AB - The generation of a wide ESTs library and database from Trichoderma harzianum CECT 2413 was the base for identifying the gene ThPTR2, coding for a PTR family di/tri-peptide transporter. The deduced protein sequence of the ThPTR2 gene showed the conserved motifs and also the 12 transmembrane domains typical of the PTR transporters. The highest level of ThPTR2 expression was found when the fungus was grown in chitin as sole carbon source. We also found that ThPTR2 expression was increased when Trichoderma interacted directly in solid medium with the plant-pathogenic fungus Botrytis cinerea, showing that ThPTR2 is involved in the mycoparasitic process. Additionally, its expression was triggered by nitrogen starvation and a higher level of expression was also found when Trichoderma was grown in secondary nitrogen sources like allantoin, yeast extract, and urea. However, no difference was found when Trichoderma was grown in presence or absence of glucose as carbon source. Strain T34-15, a transformant that overexpressed the ThPTR2 gene, showed about a 2-fold increase in the uptake of the dipeptide Leu-Leu. Additionally, two transformants from the strain Trichoderma longibrachiatum T52 that overexpressed ThPTR2 were also studied, confirming the role of this gene in peptide transport. Other homologous genes to ThPTR2 were identified in other Trichoderma strains. ThPTR2 is the first experimentally confirmed PTR family transporter gene from filamentous fungi. PMID- 16466954 TI - Cloning and characterization of the erg1 gene of Trichoderma harzianum: effect of the erg1 silencing on ergosterol biosynthesis and resistance to terbinafine. AB - Trichoderma species are commonly used as biocontrol agents of different plant pathogenic fungi. Terpene compounds are involved in the biocontrol process due to their antifungal properties (e.g., ergokonins and viridins) but additionally their structural function in the cell membranes (ergosterol) is essential. We report here the characterization of the T. harzianum erg1 gene, encoding a squalene epoxidase, a key enzyme in the biosynthesis of triterpene derivatives such as ergosterol. In T. harzianum the partial silencing of the erg1 gene gave rise to transformants with a higher level of sensitivity to terbinafine, an antifungal compound that acts specifically over the squalene epoxidase activity. In addition, these silenced transformants produced lower levels of ergosterol than the wild type strain. Finally, the silencing of the erg1 gene resulted in an increase in the expression level of the erg7 gene that encodes the oxidosqualene lanosterol-cyclase, another enzyme of the terpene biosynthesis pathway. PMID- 16466955 TI - Turning up the heat: the effects of thermal acclimation on the kinetics of hsp70 gene expression in the eurythermal goby, Gillichthys mirabilis. AB - Most organisms respond to temperature fluctuations by altering the expression of an evolutionarily conserved family of proteins known as heat shock proteins (Hsps). Studies have shown Hsp expression and the activation of HSF1, one of the primary regulators of Hsp transcription, are highly malleable, varying with the recent thermal history of the organism; however, the mechanisms that confer plasticity to the regulation of this ubiquitous response are not well-understood. This study furthers our knowledge in this area by characterizing the activation kinetics of HSF1 and the corresponding transcription of hsp70 in the liver of the eurythermal goby, Gillichthys mirabilis, following a month-long acclimation at 13, 21 or 28 degrees C. Our data revealed HSF1 DNA-binding kinetics varied as a function of acclimation temperature and magnitude/duration of exposure, with gobies acclimated at 21 degrees C exhibiting the most robust response. Hsp70 mRNA followed a similar pattern with induction first occurring in the 13 and 21 degrees C fish, and then most robustly in the 28 degrees C group at 36 degrees C. The hsp70 mRNA induction pattern was corroborated by levels of HSF1 DNA-binding activity in each group and may have been lowest in the 28 degrees C group due to the 2-fold greater levels of hsp70 protein prior to thermal exposure. This study illustrates the integral role of HSF1 as a key regulator of Hsp induction and helps explain the plasticity of this response in ectothermic organisms. PMID- 16466956 TI - Novel role for the orphan nuclear receptor Dax1 in embryogenesis, different from steroidogenesis. AB - Cytomegalic adrenal hypoplasia congenita (AHC) is an X-linked disease caused by mutations in DAX1-encoding gene NR0B1, previously thought to function primarily in steroidogenesis. We sought to determine the expression pattern for Dax1 along with known network partners in early embryogenesis and to determine a steroidogenic capacity for the embryo prior to the establishment of the urogenital ridge at embryonic day 9 (E9). Here, we report that murine Dax1 is a unique marker in early embryonic development, distinguishing the extraembryonic (proximal) endoderm from the remainder of the developing embryo. We showed that Wilms tumor 1, steroidogenic factor 1, and estrogen receptor beta were expressed throughout the embryo, but the progesterone, estrogen alpha and androgen receptors, cytochrome P450 (Cyp11a1) and Nur77 were not observed in any of the embryonic layers. Lack of Cyp11A1 expression at this stage confirmed an absence of inherent steroidogenic capacity for the early embryo. The role of Nr0b1 in embryonic stem (ES) cells was investigated using siRNA knockdown, resulting in differentiation toward endoderm-like fate. Nr0b1 conditional knockout in ES cells led to differentiation, confirming our knockdown results. Our investigations suggest that Nr0b1 functions in a novel role in the maintenance of a relatively undifferentiated state. Our results further suggest that the failure of conventional murine Nr0b1 knockout attempts may be due to disregulated differentiation. PMID- 16466957 TI - Clinical, biochemical, and molecular findings in three patients with 3 hydroxyisobutyric aciduria. AB - 3-Hydroxyisobutyric aciduria is a rare entity and affected individuals display a range of clinical manifestations including dysmorphic features and neurodevelopmental problems in the majority of patients. Here, we present two novel patients with 3-hydroxyisobutyric aciduria. To our knowledge, these are the 11th and 12th cases of 3-hydroxyisobutyic aciduria reported. It is believed that a deficiency in 3-hydroxyisobutyrate dehydrogenase is the most likely cause of this disorder. Measurement of 3-hydroxyisobutyrate dehydrogenase activity in fibroblasts homogenates of the two newly identified patients and a previously reported patient, however, revealed similar activities as in control fibroblasts. Since other enzymes with overlapping substrate specificity could conceal abnormal 3-hydroxyisobutyrate dehydrogenase activity, we cloned a candidate human cDNA for 3-hydroxyisobutyrate dehydrogenase (HIBADH). By heterologous expression in Escherichia coli, we showed that the product of the HIBADH gene indeed displays 3 hydroxyisobutyrate dehydrogenase activity. Mutation analysis of the corresponding gene in the patients suffering from 3-hydroxyisobutyric aciduria revealed no mutations. We conclude that HIBADH is not the causative gene in 3 hydroxyisobutyric aciduria. PMID- 16466958 TI - Glutaric acidemia type 1 in patients of Lumbee heritage from North Carolina. AB - Glutaric acidemia type I (GA-I) is an autosomal recessive disorder of the catabolism of lysine, hydroxylysine, and tryptophan caused by deficiency of glutaryl-CoA dehydrogenase (GCD). Among our patients with GA-I, we noted a prevalence of Lumbee individuals. The Lumbee are a close-knit Native American tribe of eastern North Carolina. Five Lumbee individuals with GA-I had homozygous 1240G>A mutations in GCD. This is a rare, known mutation that was likely introduced by a Lumbee founder. PMID- 16466959 TI - Founder mutation causing infantile GM1-gangliosidosis in the Gypsy population. AB - The Gypsies are a trans-national founder population of Asian descent, whose genetic heritage is still incompletely characterized. Here, we describe the first founder mutation leading to a lysosomal storage disorder in this population: R59H in GLB1, which causes infantile GM1-gangliosidosis. The R59H carrier rate is approximately 2% in the general Gypsy population and approximately 10% in the Rudari sub-isolate. Haplotype analysis suggests that the Gypsy diaspora may have contributed to the spread of this mutation to South America. PMID- 16466960 TI - Fractionated photodynamic therapy for a human oral squamous cell carcinoma xenograft. AB - The aim of this study was to define the appropriate fractionation interval between photodynamic therapies (PDTs) for enhanced anti-tumour effects on human oral squamous cell carcinoma (HOSCC). Reoxygenation of HOSCC and the proliferative kinetics of the tumour cells following PDT exposure were evaluated in terms of immunohistochemical expression of vascular endothelial growth factor (VEGF) and the proliferating cell nuclear antigen (PCNA). The immunohistochemical expression of VEGF was quantitatively determined by computer-assisted image analysis. The VEGF expression and the PCNA labeling indices (LIs) of the tumour cells were assessed at varying time intervals after PDT. No significant differences were observed in PCNA LIs between the control group and experimental groups at 24, 48, and 72 h after PDT. The expression of VEGF after PDT exposure was demonstrated to be higher in the experimental group at 6 h than the control group, and then was comparable at 24 h between the both groups. These results indicate that the tumour cells surviving from PDT have proliferative potential, and that oxygenation in tumours subjected to PDT may be recovered after 24 h. In the next experiment, two protocols of laser irradiation in PDT were assessed on the basis of tumour volume between fractionated exposure with a 24-h interval and continuous exposure. Regrowth of the tumour was significantly suppressed by fractionated PDT. We propose here that fractionated light exposure with a 24-h interval should be utilized in PDT for an enhanced anti-tumour effect. PMID- 16466961 TI - Development of the first potent and specific inhibitors of endocannabinoid biosynthesis. AB - Enzymes for the biosynthesis and degradation of the endocannabinoid 2 arachidonoyl glycerol (2-AG) have been cloned and are the sn-1-selective diacylglycerol lipases alpha and beta (DAGLalpha and beta) and the monoacylglycerol lipase (MAGL), respectively. Here, we used membranes from COS cells over-expressing recombinant human DAGLalpha to screen new synthetic substances as DAGLalpha inhibitors, and cytosolic fractions from wild-type COS cells to look for MAGL inhibitors. DAGLalpha and MAGL activities were assessed by using sn-1-[14C]-oleoyl-2-arachidonoyl-glycerol and 2-[3H]-arachidonoylglycerol as substrates, respectively. We screened known compounds as well as new phosphonate derivatives of oleic acid and fluoro-phosphinoyl esters of different length. Apart from the general lipase inhibitor tetrahydrolipstatin (orlistat) (IC50 approximately 60 nM), the most potent inhibitors of DAGLalpha were O-3640 [octadec-9-enoic acid-1-(fluoro-methyl-phosphoryloxymethyl)-propylester] (IC50 = 500 nM), and O-3841 [octadec-9-enoic acid 1-methoxymethyl-2-(fluoro-methyl phosphinoyloxy)-ethyl ester] (IC50 = 160 nM). Apart from being almost inactive on MAGL, these two compounds showed high selectivity over rat liver triacylglycerol lipase, rat N-acylphosphatidyl-ethanolamine-selective phospholipase D (involved in anandamide biosynthesis), rat fatty acid amide hydrolase and human recombinant cannabinoid CB1 and CB2 receptors. Methylarachidonoyl-fluorophosphonate and the novel compound UP-101 [O-ethyl-O-p-nitro-phenyl oleylphosphonate] inhibited both DAGLalpha and MAGL with similar potencies (IC50 = 0.8-0.1 and 3.7-3.2 microM, respectively). Thus, we report the first potent and specific inhibitors of the biosynthesis of 2-AG that may be used as pharmacological tools to investigate the biological role of this endocannabinoid. PMID- 16466962 TI - Programme to improve the use of beta-blockers for heart failure in the elderly and in those with severe symptoms: results of the BRING-UP 2 Study. AB - BACKGROUND: Beta-blockers are underused in HF patients, thus strategies to implement their use are needed. OBJECTIVES: To improve beta-blocker use in elderly and/or patients with severe heart failure (HF) and to evaluate safety and outcome. METHODS: Patients with symptomatic HF and age>/=70 years or left ventricular EF<25% and symptoms at rest were enrolled, including those already on beta-blocker treatment. Patients who were not receiving a beta-blocker were considered for carvedilol treatment. All patients were followed up for 1-year. RESULTS: Of the 1518 elderly patients, 505 were already on beta-blockers, and carvedilol was newly prescribed in 419 patients. At 1-year, patients treated with carvedilol had a lower incidence of death [10.8% vs. 18.0% in already treated (adjusted RR 0.68; 95%CI 0.49-0.96) and 11.2% in newly treated patients (adjusted RR 0.68; 95%CI 0.48-0.97)]. Of the 709 patients with severe HF, 38.4% were already on beta-blockers, and carvedilol was newly prescribed in 189 patients. Patients not treated with carvedilol showed the worst clinical outcome. Total rate of discontinuation (including adverse reaction and non-compliance) was 14% and 9%, respectively, in elderly and severe patients. CONCLUSIONS: In a real world setting, beta-blocker treatment was not associated with an increased risk of adverse events in elderly and severe HF patients. PMID- 16466963 TI - The effect of aortic valve replacement on plasma B-type natriuretic peptide in patients with severe aortic stenosis--one year follow-up. AB - BACKGROUND: B-type natriuretic peptide (BNP) is synthesized in cardiac tissue in response to increased wall stress and myocardial hypertrophy. AIMS: In patients with severe aortic stenosis (AS) we examined the effect of aortic valve replacement (AVR) on plasma BNP and association between BNP and left ventricular mass index (LVMI) preoperatively and in the reverse-remodeling phase twelve months postoperatively. We also examined the correlation between BNP and NYHA class and between BNP and age. METHODS AND RESULTS: Plasma BNP analyses and echocardiographic measurements were performed preoperatively, before discharge after AVR, and at twelve months in twenty-two patients. BNP was additionally measured at six months. Preoperatively, BNP was 283+/-45 pg/ml (mean+/-SEM). Following an immediate postoperative increase (441+/-38 pg/ml), BNP values decreased towards normal values at six and twelve months (139+/-25 and 130+/-18 pg/ml, respectively). LVMI was 206.5+/-15.8 g/m(2) preoperatively and decreased to 119.7+/-7.2 g/m(2) at twelve months with a correlation between LVMI and BNP preoperatively only (r=0.45, p<0.05). There was no correlation between BNP and NYHA-class, whereas BNP correlated to age both pre- and post-operatively. CONCLUSION: We report an increase in plasma BNP in patients with AS. Following a further transient increase postoperatively, BNP levels decreased at six and twelve months after AVR. BNP correlated with LVMI preoperatively, and with age both preoperatively and at twelve months. PMID- 16466964 TI - RNA interference in cancer. AB - In the recent years, RNA interference (RNAi) has emerged as a major regulatory mechanism in eukaryotic gene expression. The realization that changes in the levels of microRNAs are directly associated with cancer led to the recognition of a new class of tumor suppressors and oncogenes. Moreover, RNAi has been turned into a potent tool for artificially modulating gene expression through the introduction of short interfering RNAs. A plethora of individual inhibitory RNAs as well as several large collections of these reagents have been generated. The systems for stable and regulated expression of these molecules emerged as well. These tools have helped to delineate the roles of various cellular factors in oncogenesis and tumor suppression and laid the foundation for new approaches in gene discovery. Furthermore, successful inhibition of tumor cell growth by RNAi aimed at oncogenes in vitro and in vivo supports the enthusiasm for potential therapeutic applications of this technique. In this article we review the evidence of microRNA involvement in cancer, the use of short interfering RNAs in forward and reverse genetics of this disease, and as well as both the benefits and limitations of experimental RNAi. PMID- 16466965 TI - Targeted cDNA differential display (TcDD). AB - Targeted cDNA differential display (TcDD) was developed to study expression of a different selected gene families especially those at low copy numbers per cell. This method is an adaptation of our previously described targeted genomic differential display method (TGDD). In TcDD, the expression of genes containing target sequences such as CAG repeating sequences or genes encoding for zinc finger binding proteins were followed in an experimental rat model with salt induced hypertension. DNA sequencing experiments demonstrated that the effectiveness of targeting was greater than 99%. PMID- 16466966 TI - Risk factors for depression in patients with epilepsy. AB - PURPOSE: Symptoms of depression are present in 40 to 60 percent of patients with epilepsy. Prior research indicated significant correlation between the incidence and frequency of focal seizures and clinical depression, especially in patients with temporal lobe epilepsy. Anticonvulsive drugs and psychosocial factors contribute to the occurrence of depression as well. The aim of the study was to determine the major depression risk factors in patients with epilepsy. METHODS: The research was conducted on 203 patients with epilepsy (117 females and 86 males), aged 18 to 50 years, with total time of illness ranging from 60 to 580 months. All subjects underwent the same research protocol, which was applied interictally. Interictal depression was diagnosed according to ICD-10 diagnostic criteria for affective and delusional disorders. The diagnosis was supported by Beck Depression Inventory (BDI), Hamilton Depression Rating Scale (HAM-D) and Montgomery-Asberg Depression Rating Scale (MADRS). Statistical analysis included chi2 test, Fisher's exact test and stepwise logical regression model analysis. RESULTS: In our study 100 patients with epilepsy out of 203 suffered from concurrent depression (49.2%); 76 of them had severe depression (37.4%) and 24 patients had mild depression (11.8%). Complex partial seizures and absence of secondary generalized tonic-clonic seizures were found to be the risk factors for depression. Treatment with clonazepam, frequent hospitalizations (drug resistancy) and lack of occupational activity were revealed to be additional significant contributing factors. CONCLUSIONS: Depression in patients with epilepsy is a serious medical and social problem since it afflicts almost one half of all patients treated in epilepsy referral centers. It seems to be correlated with certain types of epileptic seizures, with high frequency of seizures, sub-optimal pharmacologic treatment and lack of occupational and social activity. PMID- 16466967 TI - Effect of the flavonol quercetin on ion transport in the isolated intestine of the eel, Anguilla anguilla. AB - Flavonoids are phenolic compounds used in fish diet formulations for the control of sex differentiation. Consequently it is of interest to know their effects on fish gastrointestinal mucosa that is the first target of these substances after food intake. We studied the effects of the flavonol quercetin on the transepithelial electrical parameters of the isolated intestine of the eel, Anguilla anguilla, by employing Ussing chamber technique. We showed that luminal quercetin (10(-4) M) reduced the serosa negative I(sc) (short circuit current) due to a net Cl- absorption in the control conditions. The reduction of I(sc) was blocked by luminal glybenclamide (10(-4) M). The I(sc) response to the flavonoid was also observed when HCO3- was omitted from the perfusion solutions. The effect of quercetin was nullified by TMB8, a blocker of Ca2+ release from intracellular stores, and was strongly reduced by trifluoroperazine, an inhibitor of calmodulin actions, while it was unmodified by experimental manoeuvres able to alter cytosolic cAMP concentrations. These results suggest that mucosal quercetin stimulated Cl- but not HCO3- secretion; the secretory activity appears to be dependent on the Ca2+-calmodulin system, independent of the cAMP pathway. In addition, this study showed that flavonol mimics the effect of carbachol on the tissue. This conclusion is supported by the following observations: 1) noradrenalin reversed the reduction of I(sc) produced by both carbachol and quercetin; 2) the flavonol was ineffective in tissues in which the muscarinic agonist had already elicited its maximal reduction of I(sc). PMID- 16466968 TI - Rates of cortisol increase and decrease in channel catfish and sunshine bass exposed to an acute confinement stressor. AB - Channel catfish and sunshine bass were exposed to a low-water stress event and allowed to recover in fresh water or a solution of metomidate (dl-1-(1 phenylethyl)-5-(metoxycarbonyl) imidazole hydrochloride), which inhibits the synthesis of cortisol. Change in time of plasma cortisol was used as an index of cortisol secretion and clearance. Plasma cortisol and glucose increased during the exposure to low-water stress in both fish, but the changes of both plasma components were more dramatic in sunshine bass. Exposure to metomidate during recovery resulted in a short-term increase in plasma glucose but differences between controls and metomidate-exposed fish were relatively minor thereafter. Cortisol began to decrease in catfish immediately after the removal of the stress but continued to increase for 15 min in sunshine bass recovering in fresh water and for 5 min in bass recovering in metomidate. Catfish recovering in fresh water had a cortisol elimination rate of -1.28 ng/mL/min compared with -2.45 ng/mL/min for fish recovering in metomidate (P>0.05) while sunshine bass recovering in fresh water had an elimination rate of -6.96 ng/mL/min compared with -4.50 ng/mL/min for fish recovering in metomidate (P>0.05). These data indicate that the rapid decrease of plasma cortisol after removal of the stressor is due to an almost immediate decrease of secretion, tissue uptake and a rapid renal loss due to the absence of a plasma binding protein. PMID- 16466969 TI - Differential effects of cervical vagosympathetic and mediastinal nerve activation on atrial arrhythmia formation in dogs. AB - To investigate the influence of the thoracic autonomic neuronal hierarchy on atrial arrhythmia formation, we compared the characteristics of atrial tachyarrhythmias induced by electrical stimulation of 1) the right vagosympathetic nerve complex at the cervical level and 2) the more caudal juxta cardiac mediastinal nerves located on the anterior surface of the superior vena cava. Unipolar electrograms were recorded from 191 sites on the entire epicardial atrial surface and, in some experiments, from 63 right atrial endocardial sites. The sites of origin of initial beats at the onset of atrial tachyarrhythmias so induced were investigated analysing atrial activation maps. Neural effects on repolarization were determined by computing the integral surface subtended by unipolar recordings under basal conditions and at maximum neurally induced bradycardia, and calculating differences at each recording site. The mean area affected by nerve stimulation in all animals was significantly greater in response to vagosympathetic than mediastinal nerve stimulation. Atrial cycle length prolongation prior to tachyarrhythmia onset was more pronounced in response to vagosympathetic than mediastinal nerve stimulation. The earliest epicardial activations in early tachyarrhythmia beats were localized in the right atrial free wall and Bachmann bundle region in both cases, but with a higher incidence of double breakthroughs from septal sites of origin in response to vagosympathetic versus mediastinal nerve stimulation. Sites of early activation were associated with the areas of neurally induced repolarization changes. Thus, differential contributions are made to the electrophysiologic substrate of neurally induced atrial tachyarrhythmias depending on the pattern of engagement of neural elements within the autonomic neuronal hierarchy. PMID- 16466970 TI - Expression analysis of the Epha1 receptor tyrosine kinase and its high-affinity ligands Efna1 and Efna3 during early mouse development. AB - Interaction of Eph receptor tyrosine kinases with their membrane bound ephrin ligands initiates bidirectional signaling events that regulate cell migratory and adhesive behavior. Whole-mount in situ hybridization revealed overlapping expression of the Epha1 receptor and its high-affinity ligands ephrin A1 (Efna1) and ephrin A3 (Efna3) in the primitive streak and the posterior paraxial mesoderm during early mouse development. These results show complex and dynamic expression for all three genes with expression domains that are successively complementary, overlapping, and divergent. PMID- 16466971 TI - Novel lethal mouse mutants produced in balancer chromosome screens. AB - Mutagenesis screens are a valuable method to identify genes that are required for normal development. Previous mouse mutagenesis screens for lethal mutations were targeted at specific time points or for developmental processes. Here we present the results of lethal mutant isolation from two mutagenesis screens that use balancer chromosomes. One screen was localized to mouse chromosome 4, between the STS markers D4Mit281 and D4Mit51. The second screen covered the region between Trp53 and Wnt3 on mouse chromosome 11. These screens identified all lethal mutations in the balancer regions, without bias towards any phenotype or stage of death. We have isolated 19 lethal lines on mouse chromosome 4, and 59 lethal lines on chromosome 11, many of which are distinct from previous mutants that map to these regions of the genome. We have characterized the mutant lines to determine the time of death, and performed a pair-wise complementation cross to determine if the mutations are allelic. Our data suggest that the majority of mouse lethal mutations die during mid-gestation, after uterine implantation, with a variety of defects in gastrulation, heart, neural tube, vascular, or placental development. This initial group of mutants provides a functional annotation of mouse chromosomes 4 and 11, and indicates that many novel developmental phenotypes can be quickly isolated in defined genomic intervals through balancer chromosome mutagenesis screens. PMID- 16466972 TI - Postnatal development of carotid body glomus cell response to hypoxia. AB - This study examines developmental changes in CB glomus cell depolarization, intracellular calcium ([Ca(2+)](i)) and the magnitude of an O(2)-sensitive background ionic conductance that may play roles in the postnatal increase in oxygen sensitivity of glomus cells isolated from rats of 1-3 days and 11-14 days postnatal age. Using fura-2 and perforated patch whole cell recordings, we simultaneously measured [Ca(2+)](i) and membrane potential (E(m)) during normoxia and hypoxia. Resting E(m) in normoxia was similar at both ages. Hypoxia caused a larger E(m) depolarization and correspondingly larger [Ca(2+)](i) response in glomus cells from 11- to 14-day-old rats compared to 1-3-day-old rats. E(m) and [Ca(2+)](i) responses to 40mM K(+) were identical between the two age groups. Under normoxic conditions both age groups had similar background conductances. Under anoxic conditions (at resting membrane potential) background K(+) conductance decreased significantly more in cells from 11- to 14-day-old rats compared to cells from 1- to 3-day-old rats. Glomus cells from newborns therefore have less O(2)-sensitive background K(+) conductance. These results support the hypothesis that postnatal maturation of glomus cell O(2) sensitivity involves developmental regulation of the expression and/or O(2)-sensitivity of background ionic conductances. PMID- 16466973 TI - Myosin heavy chain expression and muscle adaptation to chronic oral breathing in rat. AB - The purpose of this study was to establish if early chronic oral breathing could induce an ultra-structural adaptation of the diaphragm and orofacial muscles related to oral or nasal breathing. Therefore, we performed a bilateral nasal obstruction at day 8 on rat pups and the myosin heavy chain (MHC) composition of the muscles was analyzed at day 21. Nasal obstruction and the related switch to chronic oral breathing were associated with impaired growth, atrophy of olfactory bulbs, hypertrophy of adrenal glands and reduced muscle growth for all muscles studied except the diaphragm. Furthermore, we detected a smaller decrease of MHC 2b compared to MHC 2a and 2x in levator nasolabialis, a muscle involved with nasal breathing. In masseter superficialis and anterior digastric involved with oral breathing, we observed a smaller decrease of MHC 2a compared to MHC 2b or 2x, respectively. No difference was detected in the diaphragm MHC expression of oral breathing animals. Since the relative expression of fatigue resistant MHC fiber types increased in muscles involved with oral breathing, orofacial muscles seem to present a profile in MHC adapted to the transition from nasal to oral breathing, facilitating respiration. PMID- 16466974 TI - Bone stress injuries of the talus in military recruits. AB - The purpose of the present study was to assess the incidence, anatomic distribution, and nature of fatigue bone stress injuries of the talus in military recruits based on magnetic resonance imaging (MRI). Military recruits referred to MRI examination due to exercise-induced ankle and/or foot pain were identified from the MRI archives. MR images of cases with bone stress injury findings in the talus were retrospectively re-evaluated concerning the anatomic location and type of the bone stress injury. During 96 months, fifty-one consecutive recruits displayed bone stress injuries of the talus in the population base of 117,149 person-years, yielding an incidence of 4.4 (3.2-5.5)/10,000 person-years. Bilateral injuries were seen in five of the patients. Of the 56 bone stress injuries, 40 occurred in the head, 15 in the body, and 5 in the posterior part of the talus. In four cases, both the head and the body were affected. Solitary, the talus was affected in 12 cases. In 44 cases, a stress injury was also present in other tarsal bones. Assessing the severity of the bone stress injury, a grade I III injury was found in 46 and a grade IV injury with a fracture line in 10 of the cases. Injuries of the upper part of the body were associated with calcaneal stress injuries in 78% of the cases (P=0.03), and injuries of the head of the talus were associated with stress injuries in the naviculare in 60% of the cases (P=0.04). Age, sex, height, weight, body mass index, or physical fitness failed to reach statistical significance as risk factors for fatigue bone stress injuries of the talus. On MRI, the majority of the bone stress injuries of the talus were revealed in the head. A grade IV injury was discovered in 18% of the cases; in the remaining 82%, only grade I-III injuries were ascertained. In all locations, the lower grade bone stress injuries dominated. This study established the incidence of fatigue bone stress injury of the talus and indicated that these injuries are rare but not unseen in military recruits. PMID- 16466975 TI - Re: "Elevated serum homocysteine and McKusick's hypothesis of a disturbed collagen cross-linking: What do we really know?". PMID- 16466977 TI - Condylar fracture repair: use of the endoscope to advance traditional treatment philosophy. AB - Traditional treatment of subcondylar fractures with maxillomandibular fixation often results in a malreduction and significant functional and aesthetic sequelae, including facial asymmetry, decreased jaw opening, and potential for late derangements of the temporomandibular joint. When used selectively, based on preoperative CT scans, the endoscopic technique will reliably restore condylar anatomy in nearly 95% of patients, thus obviating the consequences of malunion. Furthermore, unlike traditional open techniques no significant facial scarring or permanent facial nerve palsies have resulted. Thus, the authors strongly advocate endoscopic repair of adult condylar neck and subcondylar fractures that demonstrate displacement or dislocation and have adequate proximal bone stock to accept miniplate fixation. PMID- 16466978 TI - Endoscopic repair of orbital floor fractures. AB - Significant orbital trauma can result in orbital floor fractures with subsequent prolapse of the orbital contents into the paranasal sinuses. Prolapse of the periorbita can result in extraocular muscle entrapment, diplopia, enophthalmos, and even visual loss. Management of orbital floor fractures traditionally has been accomplished through transconjunctival and subciliary incisions. These approaches provide adequate visualization and cosmetically acceptable scars. Unfortunately, post-operative lid malposition can occur in a small percentage of cases. Another limitation is easy visibility of the posterior orbit, which often is obscured by prolapsed orbital fat. In light of these limitations, some surgeons have begun to evaluate an endoscopic approach to orbital floor fractures. The endoscopic approach offers a hidden incision, improved fracture visualization, and avoidance of post-operative eyelid malposition. This article reviews the indications, technique, and potential complications of endoscopic orbital blow-out fracture repair. PMID- 16466979 TI - Endoscopic approach to medial orbital wall fractures. AB - Repair of medial orbital wall fractures can be challenging with traditional open techniques. This article describes different endoscopic-assisted approaches transcaruncular and intranasal-which have been used to successfully repair these fractures. PMID- 16466980 TI - Endoscopic repair of anterior table--frontal sinus fractures. AB - Frontal sinus fractures account for 5% to 15% of all maxillofacial injuries. Historically, a large percentage of these injuries were treated aggressively because of the long term risk of mucocele formation. This required a coronal incision with the associated surgical sequelae including a large scar, alopecia, and paresthesias. In light of these sequelae and recent advances in CT diagnosis and endoscopic treatment of mucoceles, some surgeons are starting to manage isolated anterior table fractures through an endoscopic approach. The endoscopic repair significantly reduces patient morbidity because it requires only 2 small incisions behind the frontal hairline. The endoscopic approach can be divided into two types: acute fracture reduction (covered elsewhere in this issue) and fracture camouflage. This article reviews the indications, techniques, and advantages of frontal sinus fracture camouflage. PMID- 16466981 TI - Endoscopic management of frontal sinus fractures. AB - Endoscopes have had a profound effect on nearly every surgical specialty over the past 20 years. Using endoscopic approaches, excellent visualization of the surgical site can be achieved while avoiding extensive external incisions, thus, dramatically reducing morbidity compared with traditional surgical approaches. This article outlines the state of the art with regard to the use of endoscopes for managing frontal sinus fractures, which are one of the most common fractures treated with endoscopic techniques. PMID- 16466982 TI - The rationale and technique of endoscopic approach to the zygomatic arch in facial trauma. AB - The reliable form and strategic position of the zygomatic arch make it a valuable landmark in midfacial trauma management. The benefits of arch repair have been used infrequently, mainly because traditional coronal access to this structure is fraught with undesirable sequelae. Endoscope-assisted zygomatic arch realignment and fixation allow anatomic repair without sustaining the drawbacks of extensive access incisions. The relative importance of this approach increases with trauma complexity, being most useful in Le Fort III and complex zygoma injuries. PMID- 16466983 TI - Endoscopic approach for mandibular orthognathic surgery. AB - The field of minimally invasive surgery is defined as the combination of surgical innovation with modern technology. This article describes the history of surgery and newer developments in endoscopic surgery for mandibular orthognathic surgery. PMID- 16466984 TI - Endoscopic approaches to maxillary orthognathic surgery. AB - Endoscopically assisted surgery has become an essential component in many fields of surgical specialties. The implementation of this technique to craniofacial and maxillofacial surgery is a recent development. Endoscopic approach to subcondylar mandible fractures has been established as reliable surgical method. PMID- 16466985 TI - Recombinant antisense C-myc adenovirus increase in vitro sensitivity of osteosarcoma MG-63 cells to cisplatin. AB - C-myc is an oncogene with the important role of cell proliferation controller. It has been found to be amplified and overexpressed in osteosarcoma. Moreover, it can promote cell transformation and induce metastatic features. Some studies showed that overexpression of c-myc could induce resistance in response to antineoplastic agents. Currently, we constructed the recombinant adenovirus (Ad Asc-myc) encoding antisense c-myc fragment and investigated its effect on the in vitro sensitivity of osteosarcoma MG-63 cells to cisplatin(CDDP). The osteosarcoma MG-63 cells were transfected by the Ad-Asc-myc in vitro, and Western Blot, MTT assay, RT-PCR, flow cytometry (FCM), and transmission electron microscopy (TEM) were used to study expression of c-myc and caspase-3 protein, tumor cell proliferation in vitro, cell apoptotic morphology and cell cycle change. Ad-Asc-myc encoding antisense c-myc fragment was obtained with the titer of 2.0 x 10(9) pfu/ml. Ad-Asc-myc downregulated the expression of c-myc protein after transfected MG-63 cells for 48 hours, combined with the treatment of 2.0, 5.0 microg/ml cisplatin for 2 hours can inhibited tumor cells proliferation in vitro by 33.4 and 54.2 percent, respectively, which had significant difference compared with control recombinant adenovirus (Ad-LacZ) groups (P < 0.05). RT-PCR revealed that Ad-Asc-myc downregulated expression of bcl-2 and upregulated expression of Bax, and no appreciable changes were observed in the expression of E2F-1. Detection of caspase-3 protein TEM, and FCM analysis showed that Ad-Asc myc could induce apoptosis of transfected cells, which was enhanced by the treatment of cisplatin. Cell cycle analysis showed that obvious G(2)/M phase arrested in transfected cells. In conclusion, Ad-Asc-myc increased the in vitro sensitivity of osteosarcoma MG-63 cells to cisplatin as well as induced apoptosis. PMID- 16466986 TI - A phase I study of eniluracil/5-FU in combination with radiation therapy for potentially resectable and/or unresectable cancer of the pancreas and distal biliary tract. AB - PURPOSE: Eniluracil is an effective inactivator of dihydropyrimidine dehydrogenase (DPD). It allows for oral dosing of 5-fluorouracil (5-FU), which may potentially improve the antitumor activity of 5-FU when delivered concurrently with radiotherapy while avoiding the inconvenience and morbidity of continuous infusion (CI) 5-FU. We addressed the safety of oral eniluracil/5-FU combined with radiation therapy and determined the profile of dose-limiting toxicities and recommended Phase II dose (RPTD) in patients with pancreatic and hepatobiliary cancers. METHODS AND MATERIALS: Patients with resectable or locally advanced pancreatic and biliary cancer received eniluracil (starting at 6.0 mg/m(2) q12h)/5-FU (starting at 0.6 mg/m(2) q12h). Eniluracil/5-FU were given concurrently with preoperative radiation to 4500 cGy followed by 540 cGy by reduced fields. Surgery was considered 4 weeks after completion of therapy. RESULTS: Thirteen patients were enrolled. Chemoradiotherapy was completed in all patients. The MTD was not reached and, thus, the RPTD of eniluracil/5-FU was determined to be 10 mg/m(2) q12h/1 mg/m(2) q12h. Two patients with locally advanced disease had a 30-45 percent cross-sectional tumor reduction, one of which underwent margin-negative resection. Two of 5 patients with pancreatic cancer, and 1 of 3 patients with cholangiocarcinoma, with underwent exploratory surgery had margin-negative resections. One patient had a pathologic complete response (pCR). Patient 5-FU plasma exposure increased slightly from Day 8 to Day 31. CONCLUSION: Preoperative chemoradiation with oral eniluracil/5-FU is feasible, well tolerated, and potentially effective in the neoadjuvant setting. Further investigation of oral fluoropyrimidines as radiosensitizers for pancreaticobiliary malignancies is warranted. PMID- 16466987 TI - Lipid peroxidation and antioxidant status in stomach cancer. AB - BACKGROUND: Considerable evidences have linked oxidative damage and cancer. In this article, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSHPx) activities, and malondialdehyde (MDA) and nitric oxide metabolites' levels (NO(x)) were investigated in patients with stomach cancer. METHODS: All measurments were done by spectrophotometric techniques. RESULTS: We observed a significant decrease in the activities of SOD and CAT in tumour tissues when compared with control tissues. The different of GSHPx activities and NO metabolite' levels were not statistically significant. MDA levels were significantly increased. CONCLUSIONS: We conclude that increased MDA levels and decreased antioxidant enzyme activities can be valuable parameters in assessing the possible risk of cancer. PMID- 16466988 TI - Ifosfamide/mesna as salvage therapy in platinum pretreated ovarian cancer patients--long-term results of a phase II study. AB - PURPOSE: Salvage chemotherapy in advanced ovarian cancer is not yet standardized. PATIENTS: Twenty-one consecutive patients progressing on or relapsing after previous platinum-containing treatment were eligible for treatment with ifosfamide 5 g/m(2) infused over a 24-hour period every 3 weeks in a Phase II trial. After an initial bolus of 1 g/m(2) of mesna, mesna was applied at a dosage of 5 g/m(2) concomitantly with ifosfamide followed by additional dosages of 200 mg 3 times at 4-hour intervals after termination of the ifosfamide infusion. RESULTS: The rate of objective responses was 19 percent, with a 95%CI [5.45-41.91 percent]. One patient achieved a pathologic complete remission (pCR) and 3 patients a clinical partial remission (PR). Median time-to-progression was 3 months. One patient was a long-term survivor. Main toxicities according to NCI CTC included Grade 4 neurotoxicity in one patient, Grade 3 gastrointestinal toxicity in 5 patients, Grade 3 infection in one patient, and Grade 3 and 4 leucopenia in 6 and 2 patients, respectively. CONCLUSIONS: Monotherapy with ifosfamide represents an active regimen for salvage chemotherapy in advanced ovarian cancer patients progressing on or relapsing after previous platinum pretreatment, even yielding a long-term surivor. PMID- 16466989 TI - Inhibition of lymphokine-activated killer cells generation in vitro by soluble factors released from mixed human tumor and peripheral blood mononuclear adherent cells culture. AB - BACKGROUND: A variety of molecules produced by both tumor cells and normal cells reduce the activity of lymphokine-activated killer (LAK) cells. We tested the possible cross-regulation of mel-624 melanoma cells and adherent peripheral blood mononuclear cells (PBMCs) in affecting LAK cell activity. METHODS: PBMC adherent cells were cultured together with mel-624 melanoma cells. Supernatant was transferred to a 4-day LAK cells generation culture consisted of PBMC nonadherent cells and interleukin-2. LAK cytotoxic activity was tested in a 4-hour assay against Daudi tumor cells prelabeled with sodium (51)chromate. RESULTS: The supernatant produced within the first 48 hours of mixed mel-624 melanoma cell and adherent PBMC culture substantially (by 69 percent) reduced the generation of LAK cells, whereas the supernatant from either tumor culture or adherent PBMC culture had no effect. The inhibitory effect was manifested on the generation of LAK cells when autologous nonadherent cells were cultured with 1,000 units/ml IL-2, but there was no effect on mature LAK cell cytotoxic activity. Inhibition of LAK cell generation was partially dependent on protein synthesis and was not mediated by transforming growth factor ss (TGF-ss). CONCLUSION: Our results point toward the production of soluble, yet unidentified proteins, in mixed tumor-adherent PBMC cultures, which substantially reduced the induction of LAK cells in culture. PMID- 16466990 TI - Methylated tumor-specific DNA as a plasma biomarker in patients with glioma. AB - OBJECTIVE: Patients with systemic malignancies have substantial quantities of tumor-specific DNA in their plasma which may serve as a potential biomarker for tumor burden. This approach has not been studied in gliomas. METHODS: Methylation specific polymerase chain reaction (MSP) was used to determine the methylation status the promoters for p16/(INK4a), MGMT, p73, and RARbeta within glioma tissue and plasma. Blood was collected prior to craniotomy in 10 patients with glioma. DNA was extracted from tumor and plasma samples and assayed with MSP. Total plasma DNA also was quantified. Tumor-specific plasma DNA was defined as identification of the same methylated promoter (MP) in both tumor and plasma. RESULTS: Total plasma DNA concentration was markedly elevated (mean 6,503 ng/ml, SEM 1,400 ng/ml). Glioma tissue contained methylation of at least one promoter in 9 out of 10 (90 percent) of patients studied. Of these patients, 6 out of 9 (67 percent) demonstrated methylation of at least one of the same promoters in plasma. Five of these had one MP identified in the plasma and one had 2 MP. Overall, glioma-specific plasma DNA was present in plasma of 6 out of 10 (60 percent) of patients. Each MP DNA marker found in the plasma also was present in the intracranial tumor. CONCLUSIONS: Patients with high grade gliomas have large amounts of DNA in the plasma. Of these primary brain tumors, 90 percent contained methylated gene promoters, and in over 60 percent of these patients the same methylated promoters present in the tumor also were found in the plasma. This represents the first step to developing a quantitative plasma biomarker that could be used to monitor glioma status. PMID- 16466991 TI - Polymorphisms of CYP1A1, GSTM1, GSTT1, and prostate cancer risk in Turkish population. AB - Prostate cancer is the most common cancer among men in many countries. Although the etiology of prostate cancer largely is unknown, both genetic and environmental factors may be involved. Advanced age, androgen metabolism, and heredity-race have been reported to be possible risk factors. On the other hand, several studies indicate that genetic polymorphisms in biotransformation enzymes play a role in prostate cancer development. In this study, association of the prostate cancer risk with genotype frequencies of the Phase I (CYP1A1) and Phase II (GSTM1 and GSTT1) biotransformation enzymes was investigated in 321 Turkish individuals (152 prostate cancer patients and 169 age-matched male controls). The presence or absences of the GSTM1 and GSTT1 genes were determined by a PCR-based method. Genotypes of CYP1A1 were determined by MspI-RFLP. The prevalence of GSTM1 null genotype in the cases was 64 percent, compared to 31 percent in the control group, indicating a strong association (OR = 4.08, 95%CI = 2.50-6.69). No association was observed between either GSTT1 null genotype or CYP1A1 polymorphism and prostate cancer incidence. No statistically significant association was observed between smoking status of the patients and any of the polymorphisms studied. In conclusion, results of this study indicate that only the GSTM1 null genotype may play an important role as a risk factor for prostate cancer development in Turkish population. PMID- 16466992 TI - Intramedullary spinal cord metastases from atypical small cell lung cancer: a case report and literature review. AB - Intramedullary spinal cord metastases (ISCM) are usually the result of rapidly progressing systemic malignancy. Lung cancer represents the most common solid tumor associated with the development of ISCM. We describe a 47-year-old female with atypical small cell lung cancer (SCLC) developing ISCM. After a thoracoscopic biopsy she was treated with combination chemotherapy consolidated by mediastinal radiotherapy leading to complete remission. Three months later, she developed a Brown-Sequard syndrome and an MRI scan revealed ISCM at the T10 T12 levels, and secondary brain lesions. Despite treatment with steroids and thoracic spine/brain radiotherapy, no recovery of her motor function was seen and she died 4 months later due to progressive disease in the CNS. The present case, adds to the existing list of ISCM cases reported so far for lung cancer, undermine the ominous prognosis and limited treatment options available, and an extensive literature overview and discussion of similar cases is provided. PMID- 16466993 TI - Should continuous infusion 5-fluorouracil become the standard of care in the USA as it is in Europe? AB - The mechanism of action of 5-fluorouracil (5-FU) and its pharmacologic behavior are influenced by its mode of administration. Several clinical studies have been conducted with the purpose of evaluating the difference between the continuous (CI 5-FU) and the bolus infusion of 5-FU (BI 5-FU). We focus our review on the studies relevant to the treatment of colorectal cancer, both in the adjuvant and metastatic setting. While individual trials fail to show a survival benefit for CI 5-FU, a meta-analyses of 7 trials shows an improvement in overall survival (OS) over BI 5-FU in metastatic colorectal cancer treatment. All trials in the same setting reveal a different toxicity profile for CI 5-FU that is generally more favorable than BI 5-FU. In the adjuvant setting, CI 5-FU allows the duration of therapy to be shortened by half without compromising the efficacy. CI 5-FU is the regimen of choice when given concurrently with radiation. When given in combination with other cytotoxic agents, CI 5-FU seems to be associated with less toxicity and potentially higher efficacy. Oral fluoropyrimidines, especially capecitabine, appear to behave in similar manner to CI 5-FU and may offer a convenient alternative to the usage of infusion pumps and indwelling catheters. While clinical trials are ongoing to compare capecitabine to CI 5-FU, we believe that CI 5-FU should be offered to patients in the United States given its favorable toxicity profile and higher efficacy in several settings. PMID- 16466994 TI - The surgical management of lung cancer. AB - Since the introduction of the pneumonectomy as a technically feasible strategy for the treatment of lung cancer, surgical resection has played a pivotal role in the management of early stage non-small cell lung carcinoma (NSCLC). In the last two decades, surgical, medical, and radiation oncologists have produced a growing body of evidence to support the combination of neoadjuvant or adjuvant treatments with standard surgical resection, to improve disease-free and overall survival for specific patient subgroups. Furthermore, alternatives to aggressive surgical management have evolved for patients who are medically inoperable due to compromised pulmonary function or other comorbidities. In this review, surgical options and multimodal treatment strategies are discussed, as well as completed and ongoing clinical trials addressing the surgical management of NSCLC. PMID- 16466995 TI - "Nothing is wrong, doctor": understanding and managing denial in patients with cancer. AB - From a psychological perspective, denial is the process by which the mind defends itself against painful or threatening thoughts, feelings, perceptions, or information and may manifest in healthy or sick persons as well as in patients or those who care for them. For someone with a possible cancer diagnosis, there are several individual steps involved in the denial process, as the threatened meaning of one or more symptoms is sculpted into a more acceptable reality. Although most often felt to be a dysfunctional response, denial of illness is widespread and may serve an important adaptive or coping function, especially in a person facing the challenge of cancer. It may help a person cope with the various stages of their illness and treatment by allowing them time to process distressing information at a manageable rate. However, denial may also be dysfunctional, especially if it causes delay in seeking treatment for symptoms that present in the context of a true malignancy. Many clinicians misdiagnose denial when avoidance, disavowal, or another coping mechanism is invoked or when cognitive impairment or sensory deficits adversely affect perception. This article discusses denial in the context of cancer diagnosis, treatment, and prognosis and strategies for managing this common condition. PMID- 16466996 TI - Past, present, and future strategies in the treatment of high-grade glioma in children. AB - Unlike high-grade glioma in adults, these tumors represent a minority of all primary central nervous system neoplasms in the pediatric age group (< 22 years). Treatment is quite challenging because of the resistance of high-grade glioma in children to radiotherapy and chemotherapy. Whereas maximum resection and radiotherapy are the mainstay of treatment for this type of tumor in children > 3 years, the role of chemotherapy is less clear. Despite the use of multimodality therapy, less than 20 percent of these children are long-term survivors. This review provides an overview of relevant past, current and future treatment strategies for high-grade glioma in children. PMID- 16466997 TI - Advances in radioimmunotherapy in the age of molecular engineering and pretargeting. AB - Now that radioimmunotherapy is an approved method for the treatment of certain types of non-Hodgkin's lymphoma, investigators are turning to new approaches to further improve radionuclide targeting in hopes of expanding the use of this technology. A number of innovative recombinant proteins have been developed with more favorable pharmacokinetic and targeting properties than standard whole IgG, which conceivably could improve the therapeutic index for cancer treatment. Pretargeting methods also are coming of age, with preclinical and early clinical studies in a variety of cancers illustrating how this alternative approach can enhance the therapeutic window several-fold of what has been possible with directly radiolabeled IgG. This review will discuss some of these promising new developments. PMID- 16466998 TI - Target practice: figuring out which, when, and why to use systemic therapies for metastatic colon cancer. AB - There have been many recent advances in the field of systemic chemotherapy for metastatic colorectal cancer. Each treatment affects a different target, including thymidylate synthase, topoisomerase, DNA, the epidermal growth factor receptor, and vascular endothelial growth factor. These treatments have been used in multiple combinations, schedules, and as first-line, second-line, and third line therapies. The current clinical trials reported can help guide the clinician to determine the appropriate timing, combination, and schedule of chemotherapy agents. Although the decision regarding colorectal cancer chemotherapy is now more complicated, our patients have certainly benefited from better response rates and most importantly, longer survival times. PMID- 16467000 TI - The meaning of translation. PMID- 16467001 TI - Another example of the highly variable clinical course of recurrent/resistant/persistent ovarian cancer. PMID- 16467002 TI - The phenomenon of osteonecrosis of the jaw in patients with metastatic breast cancer. AB - Osteonecrosis of the jaw (ONJ) has been noted to occur in patients with cancer receiving bisphosphonate therapy and may also be referred to as bisphosphonate associated osteonecrosis (BON). ONJ has come to attention through case reporting and the pathogenesis and true incidence of this condition are not yet defined. ONJ appears to be a relatively uncommon event, but it is of clinical significance; the evolving data on ONJ are discussed here. PMID- 16467003 TI - Medical confidentiality and patient privacy: the Jewish perspective. AB - This article presents the approach of Jewish law to the major issue of medical confidentiality and patient privacy. For an extensive discussion of Jewish medical ethics and an in-depth presentation of the Jewish view of 39 major issues in medical ethics, the reader is referred elsewhere [15]. Briefly, the Jewish view toward medical ethical subjects is predicated on the general principle of the supreme value of human life. In Judaism, all biblical and rabbinic laws are temporarily waived in order to save a human life. Physicians are obligated to heal patients from their illness, to induce remission and cure of disease whenever possible. Similarly, patients are obligated to lead healthy lifestyles, to consult physicians when they are sick and to be compliant with the physician's therapeutic recommendations. The Jewish view on medical confidentiality and patient privacy as presented in this essay flows from these general principles of Jewish medical ethics. PMID- 16467005 TI - Nicotine and cognitive efficiency in alcoholics and illicit stimulant abusers: implications of smoking cessation for substance users in treatment. AB - Cigarette smoking is prevalent among alcoholics and illicit substance abusers. However, the potentially confounding effect of nicotine on studies of cognition in detoxified substance users has rarely been addressed. In the current study of 87 participants, behavioral and electrophysiological indices of cognitive efficiency were measured in tobacco smokers from four groups: alcoholics, illicit stimulant abusers, concurrent abusers, and control subjects. Data were collected from 2001 to 2003. We hypothesized that acute nicotine administration would modify cognitive deficits in alcoholics and illicit stimulant abusing groups. An adaptation of the Rapid Visual Information Processing task was administered after stabilization of nicotine levels via a high- or low-dose transdermal nicotine patch. Across groups, increased nicotine dose was associated with decreased reaction time (p = .03). A group x nicotine dose interaction trend was noted in which increased nicotine was associated with increased correct responding within the alcoholic group (p = .02). No significant differences in electrophysiology were observed. These results suggest that nicotine may modify cognitive efficiency in alcoholics and illicit stimulant abusers, a concept with relevance to both the design of experimental work and the treatment of alcohol and illicit stimulant dependence. Further work is needed to determine whether this effect predominantly reflects facilitation of cognition function or alleviation of nicotine withdrawal. PMID- 16467006 TI - Hospitalization due to alcohol and drug abuse in first- and second-generation immigrants: a follow-up study in Sweden. AB - The detrimental effects of substance abuse on the individual and on society are well known. Health care resources must target where they are most needed. This study investigated (1) whether the increased risk of being hospitalized for alcohol abuse and drug abuse among certain immigrant groups is present in the next generation and (2) whether having one parent born in the host country (i.e., Sweden) has a protective effect on the risk of being hospitalized for alcohol abuse and drug abuse among second-generation immigrants. In total, 2,243,546 persons aged 20-39 years were followed from 1 January 1992 until 31 December 1999 for first hospital admission due to alcohol abuse or drug abuse. A Cox regression model was used to estimate hazard ratios for each diagnosis group. When the Swedish majority population was used as reference, the highest risks of alcohol abuse and drug abuse were found among first- and second-generation Finns, after adjustment for income. Among second-generation Finns with one parent born in Sweden, the increased risk of being hospitalized for alcohol abuse and drug abuse was lower than among second-generation Finns with both parents born in Finland. Health care resources need to target certain groups of first- and second generation immigrants. PMID- 16467007 TI - Flunitrazepam abuse and malicious use in Texas, 1998-2003. AB - Flunitrazepam is a potent benzodiazepine that is subject to abuse and malicious use. This study describes the patterns of flunitrazepam abuse and malicious use calls received by Texas poison centers during 1998-2003. The distribution of calls by year of call, geographic location of caller, patient gender and age, exposure site, and medical outcome were determined. There was no clear annual trend for abuse calls, but there was a consistent decline in the number of malicious use calls. A significantly higher percentage of abuse calls originated in south and west Texas and of malicious use calls in west Texas. Most abuse patients were males (55%) and adolescents (76%), and most of the exposures occurred in patient's own residence (68%), followed by school (16%). Most of the malicious use patients were females (93%) and adults (74%), and the greatest proportion of the exposures occurred in public areas (47%), followed by the patient's own residence (26%). The highest percentage of both abuse (48%) and malicious use (55%) involved minor effects. However, malicious use calls were significantly less likely to involve no effect (2% vs. 21%) and more likely to involve moderate effects (36% vs. 23%). Reported flunitrazepam abuse and malicious use calls in Texas differed with respect to geographic location of the caller, patient gender and age, exposure site, and medical outcome. Poison centers and health care providers might want to consider these differences when targeting populations for education and prevention efforts. PMID- 16467008 TI - Cocaine addiction and family dysfunction: a case-control study in southern Brazil. AB - Family-related factors play a crucial role in the onset, development, and maintenance of drug misuse and addiction. To investigate the hypothesis that families with a cocaine addicted son display more dysfunction than control families, we applied the Personal Authority in the Family System and the Self Report Measure of Family Functioning to 67 case triads (child-father-mother) in which the son was an inpatient with a primary diagnosis of cocaine abuse or dependence and to 67 matched control triads from the community (Pelotas, Brazil) in 2000. Case children, mothers, and fathers were significantly more likely to present nuclear family triangulation than their matched control subjects. In addition, case triads had a significantly greater chance of perceiving family functioning as enmeshment. These results suggest both structural and functional differences in these families. We argue for the need of interventions in the family as a whole and for further research in this area. PMID- 16467009 TI - Racial and ethnic differences in predictors of smoking cessation. AB - Racial/ethnic differences in the determinants of smoking cessation could have important treatment implications. The current study examined racial/ethnic differences in smoking cessation, prospective predictors of cessation, and whether the predictive ability of these factors differed by race/ethnicity. Participants were 709 employed adults recruited through the National Rural Electric Co-op Association or through natural gas pipeline corporations. Data were collected in 1990 and 1994. Although race/ethnicity was not predictive of abstinence, Hispanic, African American, and White smokers displayed differential on tobacco-, alcohol-, and work-related variables. These racial/ethnic differences highlight the specific factors that should be considered when providing smoking cessation treatment to specific populations. Limitations are noted. PMID- 16467010 TI - Epidemic of HIV coupled with hepatitis C virus among injecting drug users of Himalayan West Bengal, Eastern India, Bordering Nepal, Bhutan, and Bangladesh. AB - A study was conducted in June 2004 to find out the epidemiology of HIV infection among injecting drug users (IDUs) of Darjeeling District of West Bengal, eastern India. The district headquarter, Darjeeling town, also known as "Queen of Hills," is a beautiful spot situated in Himalayan West Bengal that attracts a large number of tourists each year from all over the world. Another unique feature of the district is that it has international boundaries with three countries, Nepal, Bhutan, and Bangladesh. Siliguri, the part of the district on plains, acts as a transit station for these countries as well as to the entire Himalayan region of West Bengal and neighboring state, Sikkim. It is also a transit point to all northeastern states of India: Assam, Arunachal Pradesh, Nagaland, Manipur, Mizoram, Meghalaya, and Tripura. Two hundred twenty-eight study subjects (IDUs) were included in this community-based cross-sectional study from all four subdivisions of the district. Informed consent was obtained, and then personal interviews, followed by blood testing were performed using unlinked anonymous procedure. The study revealed that overall HIV seroprevalence among IDUs was 11.8% (n = 27; 95% confidence interval, 7.9-16.7), whereas seroprevalence of hepatitis C was found to be 47.7% (n = 97). Prevalence of HIV was higher in subjects from hill districts (13.5%) compared with subjects from the plains (9.2%). It also revealed that most IDUs (75.3%) used "brown sugar," an impure form of heroin, as their major addictive substance followed by injection norphine. Sharing of injecting equipment was found to be as high as 67% among IDUs, and sharing of drugs from common ampules was found to be 35.5% of the studied subjects (n = 93). Most subjects (96%) were found to clean their injecting paraphernalia with plain water. Most IDUs (98%) were found to inject intravenously. About 52% of IDUs visited sex workers one or more times within the last 1 year, and 15% of the interviewed subjects (n = 93) reported to suffer from sexually transmitted diseases during the same period. All the IDUs knew about HIV/AIDS. About 69% of the subjects knew that apparently healthy looking person might have HIV infection. HIV was found to be associated significantly with age of the injectors and duration of injecting practices. The study revealed the epidemic of HIV and hepatitis C among IDU populations at this bordering district of West Bengal for the first time that requires urgent intervention at local, national, and international levels. PMID- 16467011 TI - From the laboratory to real life: a pilot study of an expectancy challenge with "heavy drinking" young people on holiday. AB - The Alcohol Expectancy Challenge (EC) is a promising program for changing alcohol expectancies and reducing alcohol consumption in "heavy drinking" young men in a bar-lab setting. In this study the EC was adapted for use in mixed-gender groups in a holiday setting and its feasibility tested in camping resorts in the Netherlands where a lot of binge drinking takes place (summer 2002). Male and female participants (N = 170; mean age, 18.8 years) were randomly assigned to an EC or to an assessment-only control group. One day before the intervention, alcohol expectancies were measured by a Visual Analogue Scale of arousal-sedation expectancies (VAS expectancies questionnaire). At the same time, alcohol use in everyday life and on holiday was assessed by a General Drinking Questionnaire and a 24-hour drinking diary, respectively. Twenty-four hours after the intervention, the VAS expectancies questionnaire was administered again and alcohol use over the previous 24 hours was reported in the drinking diary. Six weeks after the intervention, participants were telephoned and administered oral versions of the VAS expectancies questionnaire and General Drinking Questionnaire. Data were analyzed using mixed ANOVAs. Although the study was hampered by recruitment difficulties, the EC proved feasible in this setting, was well received by youngsters, and effects on their alcohol expectancies may have been present. No effect was found on alcohol use. In conclusion, implementation must be improved and more studies are needed to come to more definite conclusions about the value of the EC in a real-life targeted intervention. PMID- 16467012 TI - Measuring confrontation during recovery from addiction. AB - The effect of confrontation on recovery from addiction continues to be a topic of considerable debate. Although many residential treatment programs view some form of confrontation as an integral part of recovery, a number of studies have found confrontation from professional treatment staff to be counterproductive. One of the problems inherent in the current debates about confrontation is the lack of a comprehensive measure of confrontation that assesses different dimensions. This study describes the development of the "Alcohol and Drug Confrontation Subscale" (ADCS), a 72-item instrument designed to measure the quantity and frequency of confrontation that individuals receive about drug or alcohol use-related problems. Confrontation is defined as an individual being told "bad things" might happen to them if they do not make changes to address a drug or alcohol use related problem or make changes to maintain sobriety. The instrument also measures the respondents' perceptions about their relationships with confronters (three-item alpha = .79) and perceptions about the confrontational statements (three-item alpha = .63). The sample included 108 individuals entering three sober living housing organizations in Northern California between 2003 and 2005. They indicated that receiving confrontational statements about alcohol or drug use-related problems was common, especially from spouses/significant others (56% of those with significant others) and family members (60%). Participants who reported receiving more confrontation reported having more positive views about their relationships with confronters and about confrontational statements than those who received less confrontation. Additional studies are necessary to establish validity, generalize results to more diverse populations, and assess confrontation at different time points during recovery. PMID- 16467013 TI - Patient and physician characteristics in relation to clinical decision making in methadone maintenance treatment. AB - Delivery of methadone maintenance treatment (MMT) varies considerably between service providers, but the reasons for this are unclear. This two-phase study involved a controlled investigation of factors that influence clinical decision making by methadone-prescribing physicians in regard to three decision-making scenarios: (1) individuals seeking induction into MMT and existing patients seeking (2) replacement and (2) takeaway methadone doses. In phase 1, physicians (n = 17) rated the diagnostic merit of 87 patient factors for each scenario. Ratings suggested that decisions are influenced by a range of subjective and "nonmedical" patient factors (e.g., contact with drug subculture, appearance, employment status, social support, having children), in addition to more conventional information sources regarding patients' medical and treatment status (e.g., being pregnant, urinalysis evidence of opioid and poly drug use, signs of opioid withdrawal). Phase 2 (n = 296) investigated relationships between physician characteristics and responses to randomized-controlled case vignettes (decisions and confidence ratings) in which the amount and type of diagnostic and nondiagnostic patient information was controlled. Vignette responses were significantly related to physician characteristics (e.g., professional orientation, location, and experience) independent of the patient information provided. Delivery of MMT may vary due to the diversity of patient factors that influence decisions and variability between physicians in the way such information is used to form judgments. Training programs for methadone prescribers should account for these sources of potential variability in treatment management. PMID- 16467014 TI - What is behind smoking among pharmacy students: a quantitative and qualitative study from Turkey. AB - This study, performed in two phases, compared the smoking status of first- and last-year pharmacy students and identified underlying factors of smoking using both "quantitative" and "qualitative" research techniques. The quantitative phase was a cross-sectional study with 207 students (of these, 102 were first-year and 105 were last-year students). The mean age at which first-year students tried tobacco/tobacco products was 14.1 +/- 3.2 and for last-year students, 16.0 +/- 2.4. The students completed an 18-item questionnaire at the end of the spring semester in May 2004. Chi-square and t-test analyses were used for statistical comparisons. Furthermore, focus group discussion techniques were used to find out the underlying factors of smoking in the second phase. PMID- 16467015 TI - Substance use among young offenders: thrills, bad feelings, or bad behavior? AB - The relationships of conduct problems, attention deficit hyperactivity disorder (ADHD) signs, and depression to substance use were examined between 1994 and 1999 among 900 incarcerated young offenders in South Australia, 206 of whom were reassessed when later readmitted to secure care. At the first and second assessments, conduct problems, ADHD signs, and depression scores all had significant concurrent associations with a measure of recent substance use. Prospectively, there were no significant associations between depression and substance use. At the zero-order level, both ADHD signs and conduct problems predicted future substance use. ADHD signs remained significantly predictive after controlling for concurrent associations. The results lend support to the view that substance use is related to general deviance and that the arousal needs associated with increased ADHD signs increase the risk of substance use. There was no support for the view that substance use in this population is self medication in response to internalizing problems. PMID- 16467016 TI - Intermittent dosing of biologics for psoriasis. PMID- 16467017 TI - Short-term oral isotretinoin therapy does not cause clinical or subclinical peripheral neuropathy. AB - BACKGROUND: Subclinical neurophysiologic abnormalities mainly suggestive of sensory fibers dysfunction were observed in some patients after 1 and 3 months of treatment with oral acitretin. Moreover, two cases of peripheral sensory neuropathy and one of sensorimotor polyneuropathy were observed after short-term oral administration of this compound. OBJECTIVE: The objective of this prospective study was to investigate whether short-term treatment with oral isotretinoin can also affect peripheral nerve function. METHODS: Serial neurologic and neurophysiologic examinations were performed on 18 young patients with severe nodulocystic acne prior to and 1 and 3 months after the onset of oral isotretinoin treatment (1 mg/kg per day). RESULTS: Clinical neurologic examination before and under treatment disclosed no abnormalities in any of the patients. There were no significant differences between the pre- and post treatment neurophysiologic parameters. Furthermore, evaluation of the serial neurophysiologic measurements in each patient separately under oral isotretinoin treatment revealed no changes fulfilling the criteria of abnormality. CONCLUSIONS: Short-term administration of oral isotretinoin in young patients does not cause clinical or subclinical neuropathy. PMID- 16467018 TI - Clinical response in psoriasis patients discontinued from and then reinitiated on etanercept therapy. AB - BACKGROUND: Although continuous therapy with the tumor necrosis factor (TNF) antagonist, etanercept, has been shown to have a favorable benefit to risk profile in the treatment of moderate to severe plaque psoriasis, it is recognized that patients and practioners may wish for intermittent treatment should life circumstances dictate. OBJECTIVE: To evaluate safety and effect maintenance of etanercept retreatment in psoriasis. METHODS: Results of a 24-week, randomized, placebo-controlled, double-blind study were previously reported. Patients who responded at week 24 (improved ?50% in psoriasis area and severity index [PASI]) discontinued etanercept until disease relapse (loss of ?50% of week 24 PASI improvement). Patients were retreated with blinded etanercept at the originally randomized dose: 25 mg or 50 mg twice weekly (BIW) or 25 mg once weekly; original placebo patients received 25 mg BIW for the final 12 weeks of the double-blind period and were retreated with etanercept 25 mg BIW. RESULTS: Psoriasis returned gradually, without untoward events, within, on average, 3 months after etanercept discontinuation. Results after 12 weeks of retreatment were similar to those achieved after the initial 12 weeks. The major limitation of this study is that it examines only one round of discontinuation/retreatment. CONCLUSIONS: Retreatment with etanercept was effective and well tolerated in psoriasis patients. PMID- 16467019 TI - Old drug--new indication. Rifampicin in psoriasis. AB - BACKGROUND: The efficacy of traditional systemic therapies for psoriasis is limited by various side effects, toxicity, drug-drug interactions, and the need for frequent laboratory monitoring. In animal models, rifampicin causes immunosuppression and in conventional doses it suppresses the T-cell function. OBJECTIVE: To show that rifampicin has a therapeutic effect in eruptive psoriasis and to try to explain its mode of action. MATERIALS AND METHODS: A total of 76 patients (34 men and 42 women, aged between 12 and 68 years) with eruptive psoriasis were enrolled in the study. They were divided into two groups according to the evidence of a concomitant streptococcal infection. Rifampicin was administered orally in a 600 mg daily dosage for at least 60 days. Only emollients were given for topical therapy. RESULTS: A statistical (chi-squared test) analysis was done and it could be concluded that improvement in the two groups was statistically indistinguishable (p = 0.892), while comparison with the control group showed a significant difference (p = 0.00082). CONCLUSION: The results express that there is no statistically significant difference between the treating groups and the effect of rifampicin could not be related only to its antimicrobial properties. Its therapeutic effect most probably is due to its immunosuppressive properties. PMID- 16467020 TI - The negative impact of psoriasis on the workplace. AB - BACKGROUND: The health-related quality of life and economic impact of psoriasis has been documented. However, the negative impact of psoriasis on the workplace is poorly understood. AIM/METHODS: Through anonymous surveys from 30 subjects of each of three psoriasis severity groups we aim to quantify the workplace impact experienced by psoriasis patients and factors that may contribute to this impact. Data were collected using several previously validated measures including the Work Productivity Assessment Index (WPAI), SF-8, Hospital Anxiety and Depression (HADS) and past medical/psoriasis history. RESULTS: Psoriasis patients experienced negative impact at work as measured by the WPAI; this impact is directly associated with disease severity. The other instruments confirmed the negative impact of psoriasis though the degree of impact did not always correlate with disease severity or work impact. Owing to the relatively small numbers, the significance of these findings is not certain. CONCLUSIONS: This preliminary study finds that psoriasis is associated with a reduction in work productivity and social functioning; this may explain the relatively high cost of this disease on both a macro- and microeconomic level. As there was a lack of significant trends in this study, formal inferences are limited and further investigation is warranted. PMID- 16467021 TI - The cost-effectiveness and cost of treatment failures associated with systemic psoriasis therapies. AB - BACKGROUND: Psoriasis is difficult at times to treat and treatment failures are not uncommon regardless of approach. With the advent of expensive biologic therapies for psoriasis there is increasing discussion on the cost efficacy of a given systemic or biologic agent. An alternative and overlooked aspect of cost efficacy is the cost that accrues from treatment failures. METHODS: We review the literature and develop a model to analyze the cost-effectiveness and the cost of treatment failures per success for various systemic psoriasis agents using a 12 week treatment period. RESULTS: For continuous-dose agents, the cost effectiveness results are: methotrexate $623, acitretin $2729, cyclosporine $2969, nUVB $3692, PUVA $4668, etanercept $16 312, and efalizumab $17 196. The cost of expected treatment failures to achieve one success for the same agents were: methotrexate $187, cyclosporine $505, PUVA $767, nUVB $1034, acitretin $1310, etanercept $8319, and efalizumab $12 897. CONCLUSIONS: Methotrexate appears to be the most cost-effective agent for the treatment of severe psoriasis. However, greater efficacy can be achieved with cyclosporine and PUVA, albeit at a greater cost. Because of the high cost of treatment failures, access to a wide array of therapies and combination regimens should not be discouraged by physicians or insurers. PMID- 16467022 TI - Intermittent versus continuous terbinafine in the treatment of toenail onychomycosis: a randomized, double-blind comparison. AB - BACKGROUND: Terbinafine is an established drug for the treatment of toenail onychomycosis. Minimizing the total dose of terbinafine and giving it intermittently could improve tolerability as well as compliance, provided efficacy is not compromised. OBJECTIVE: Two identical trials were conducted to compare the efficacy, safety and tolerability of the current standard regimen of terbinafine 250 mg daily with a new formulation of terbinafine given intermittently for three cycles of 2 weeks of treatment (350 mg daily) followed by 2 weeks off treatment. METHODS: A total of 2005 patients with a clinical diagnosis of subungual onychomycosis of the large toenail confirmed by microscopy and culture for a dermatophyte were recruited into the two trials and treated for 12 weeks. RESULTS: Patients with onychomycosis of prolonged duration (mean 9 years) and a median nail involvement of 63% with or without spikes, lateral involvement and white superficial onychomycosis (WSO) were included in the trial. The studies found a significant difference (p<0.05) in favour of standard daily dosing with terbinafine. Response rates for the primary variable complete cure (mycological and clinical cure) were lower with the new formulation in both Trial I (-5.8%; 95% CI -11.8, 0.07) and Trial II (difference -5.9%; 95% CI -12, 0.1). Both treatments were equally well tolerated, with approximately 11% of patients in both groups reporting at least one treatment-related adverse event. CONCLUSIONS: Pulsed dosing with terbinafine did not provide any clear safety advantages and was significantly less effective. Consequently, continuous treatment with terbinafine tablets remains the optimal therapy for onychomycosis. PMID- 16467023 TI - Potassium titanyl phosphate (KTP) laser treatment for molluscum contagiosum. AB - We report a case of an immunocompromised 38-year-old Asian patient who developed several hundred atypical lesions of molluscum on her face and neck, resistant to conventional treatment. These lesions were treated successfully by potassium titanyl phosphate (KTP) laser without pigmentary disturbance. KTP laser is a novel treatment for molluscum contagiosum. PMID- 16467024 TI - Evaluation of three different strengths of trichloroacetic acid in xanthelasma palpebrarum. AB - BACKGROUND: Xanthelasma is a common cutaneous condition that presents in the periocular region. Essentially benign, treatment is of cosmetic importance. OBJECTIVE: Evaluation of varying strengths of trichloroacetic acid (TCA) in xanthelasma palpebrarum. METHODS: Three strengths of TCA were used on 51 patients randomly after categorizing their xanthelasma into papulo-nodular, flat plaques and macular lesions. The average number of sittings was calculated in each category and patients were reviewed fortnightly. RESULTS: Papulo-nodular lesions required an average of two applications with 100%, 2.67 with 70% and 4.16 with 50% TCA. Flat plaques responded to an average of 1.43, 1.50 and 3.55 sittings with 100%, 70% and 50% TCA, respectively. Macular lesions responded to only one application of all strengths of TCA applied. Eleven patients developed hypopigmentation, five had hyperpigmentation and one developed mild scarring. CONCLUSION: 100% TCA gives the best results in papulo-nodular lesions, 100% or 70% TCA give similar results in flat plaque xanthelasma and in macular lesions 50% is sufficient. Hypopigmentation is the commonest side effect, followed by hyperpigmentation. Scarring is a minor problem. PMID- 16467025 TI - Chronic cutaneous lupus flare induced by systemic 5-fluorouracil. AB - 5-Fluorouracil is an antineoplastic antimetabolite responsible for a variety of cutaneous reactions. We report a case of chronic cutaneous lupus flare related to systemic 5-fluorouracil administration for breast cancer in a patient with documented history of chronic cutaneous lupus. PMID- 16467026 TI - Diagnostic accuracy for skin cancer: comparison of general practitioner with dermatologist and dermatopathologist. PMID- 16467027 TI - Dermatomyositis-like eruption with long-term hydroxyurea. PMID- 16467033 TI - Progress and regression within primary Sjogren's syndrome. AB - The purpose of this review is to give a modern view and an update of important areas in primary Sjogren's syndrome (SS), which may be the most common of the autoimmune systemic rheumatic diseases. Interest in aspects of primary SS including clinical manifestations, pathogenesis, aetiology, treatment, prognosis, etc has increased during the past three decades, the volume of scientific papers and the number of theses being the indicators. However, only a fraction of the money that is used for research into rheumatoid arthritis (RA) is used for SS, and the statement that SS is under-diagnosed, under-treated and under-researched will still be valid for several years to come. The topics that are focused on in this review are: (a) clinical areas with subsections on signs and symptoms, terminology, predictors for development of non-Hodgkin malignant lymphoma (NHML) and prognosis, (b) treatment, (c) the Danger model (aetiopathogenesis) and (d) pathology, including immunoglobulin G4 (IgG4)-positive cells. PMID- 16467034 TI - A two-year follow-up of work capacity in early rheumatoid arthritis: a study of multidisciplinary team care with emphasis on vocational support. AB - OBJECTIVE: To explore changes in sick leave patterns and work ability in patients with early rheumatoid arthritis (RA). The patients received active team support focusing on vocational rehabilitation, in addition to treatment with disease modifying anti-rheumatic drugs (DMARDs). METHODS: This is an observational study of 110 patients with early RA aged 18-60 years and not permanently disabled. All patients were monitored regularly during a 2-year period by a team comprising a nurse, an occupational therapist, a physiotherapist, a rheumatologist, and a social worker. Intervention included work-site visits and rehabilitation meetings with the employer and the official from the local social insurance office in addition to DMARD treatment and different individual treatments, and support from the team members. RESULTS: The number of patients working full-time increased from 65 to 74 (14%), those with full-time work disability decreased from 37 to 13 (65%), and patients working part-time increased from 8 to 23 (65%). This change was already evident during the first year. CONCLUSION: Active vocational support in addition to DMARD treatment may prevent or delay work disability in patients with early RA. PMID- 16467035 TI - Hand function in women and men with early rheumatoid arthritis. A prospective study over three years (the Swedish TIRA project). AB - OBJECTIVE: To describe the course of hand function in women and men during the first 3 years after diagnosis of recent-onset rheumatoid arthritis (RA), to investigate sex differences in hand function, and to study correlations between and within hand function assessments. METHODS: A total of 276 patients (69% women) with RA of a maximal duration of 12 months were recruited to the study. Hand function was assessed by the Grip Ability Test (GAT) and Signals of Functional Impairment (SOFI). Peak and average grip force over 10 s in the right and left hand was measured by an electronic device. RESULTS: Hand function was affected at diagnosis, but had improved significantly at the 3-months' follow-up and then remained stable (but still affected) in both women and men. As assessed by SOFI, hand function was worse in men than in women, whereas women had significantly lower grip force. GAT, grip force, and SOFI correlated weakly. The average and peak values of grip force correlated strongly, as did the grip force in the right and the left hand. CONCLUSION: Hand function was profoundly affected at diagnosis of RA, but improved significantly within 3 months and remained stable (but still affected) over 3 years. As expected, women on average had significantly lower grip force than men. PMID- 16467036 TI - Lack of association of the CD14/C-159T polymorphism with susceptibility and serological activity parameters of rheumatoid arthritis. AB - OBJECTIVE: CD14, the monocyte receptor for lipopolysaccharides (LPS), is an important mediator of inflammatory processes. As the T-159C exchange in the promotor of the CD14 gene was reported to lead to enhanced CD14 expression, this could be a new susceptibility gene for rheumatoid arthritis (RA). We investigated whether this single nucleotide polymorphism (SNP) serves as a risk factor for disease development or has any influence on serological activity parameters of RA or soluble CD14 (sCD14) levels. PATIENTS AND METHODS: A total of 130 patients with RA, diagnosed according to the revised American College of Rheumatology (ACR) criteria, and 130 healthy subjects, all Caucasians, were genotyped using polymerase chain reaction (PCR). Genotype frequencies were compared by chi2 analysis. RESULTS: Forty (31%) patients vs. 39 (30%) controls were genotyped CC; 71 (55%) vs. 67 (52%) were heterozygous, and 19 (15%) vs. 24 (19%) showed the TT genotype (p = 0.7). Accordingly, the allele frequency was equally distributed (p = 0.8). There was also no significant difference in genotype distribution between subgroups of patients categorized according to serological activity parameters and sCD14 levels. CONCLUSION: We found no association between the CD14/C-159T polymorphism and increased risk for the development of RA or serological disease activity parameters or sCD14 levels. PMID- 16467037 TI - Salivary production rates fall with age in subjects having anti-centromere, anti Ro, and/or anti-La antibodies. AB - OBJECTIVES: To clarify the relationship between a subject's sicca-associated autoantibodies type and changes in salivary production rate (SPR) with age in subjects having any of these antibodies. METHODS: One hundred and eighty-five subjects (female:male = 178:7), who had at least one of the three autoantibodies, anti-centromere (ACA), anti-Ro (SSA), and/or anti-La (SSB), and 65 healthy females were enrolled. The Saxon test was used to measure SPR. RESULTS: SPRs in the seven male subjects were significantly higher than those in the 178 females tested. Therefore, only female subjects were used for the following analyses. Subjects were classified into substantially four groups according to their seropositivities for ACA, anti-Ro, and/or anti-La: group A, subjects having ACA alone; group B, subjects having anti-Ro alone; group E, subjects having anti-Ro and anti-La; group DFG, subjects having ACA with anti-Ro and/or anti-La. The frequency of Sjogren's syndrome (SS) was 74.0-94.1% in these groups. SPR did not decrease with age in normal female controls. By contrast, SPR decreased significantly with age in the groups having sicca-associated antibodies. The degree of SPR decrease compared between groups was: group A group [symbol: see text] B > group E [symbol: see text] group DFG. In the analysis of the subgroup having any of the sicca-associated antibodies but not fulfilling the classification criteria of SS, SPR also decreased with age. CONCLUSION: SPR in subjects having any of the sicca-associated antibodies (ACA, anti-Ro, or anti-La) decreased with age. PMID- 16467038 TI - Prevalence and clinical correlates of anti-phospholipid antibodies in South Africans with systemic lupus erythematosus. AB - OBJECTIVE: To determine the prevalence and clinical correlates of anti phospholipid antibodies (aPL), including anti-cardiolipin antibodies (aCL), lupus anti-coagulant (LA), anti-beta2-glycoprotein 1 (abeta2GP1), and anti-prothrombin (aPT) antibodies, in Black South African patients with systemic lupus erythematosus (SLE). METHODS: A cross-sectional study of 100 SLE patients in whom clinical characteristics, including features of the anti-phospholipid syndrome (APS), disease activity, and damage were documented, and sera tested for aCL, abeta2GP, and aPT of all isotypes, and LA. RESULTS: Positive aCL, abeta2GPI, aPT, and LA were found in 53, 84, 20, and 2 patients, respectively. Immunoglobulin (Ig)A aCL and IgG abeta2GPI were the commonest aCL (49.1%) and abeta2GPI (47%) isotypes, respectively. IgA abeta2GPI were associated with both a history of thrombosis alone (p<0.05) and a history of any clinical feature, thrombosis, and/or spontaneous abortion of the APS (p<0.05); IgA aCL were associated with a history of any clinical APS event (p<0.05); and abeta2GPI of any isotype were associated with a history of arthritis (p<0.001). CONCLUSION: Our findings provide further evidence that screening for abeta2GPI and IgA aCL isotypes may improve the risk assessment for APS in SLE patients of African extraction. Further prospective studies are warranted to determine the clinical utility of these tests and to elucidate the genetic basis for the increased IgA aPL response in SLE patients of African extraction. PMID- 16467039 TI - Low-dose intravenous cyclophosphamide in systemic sclerosis: an open prospective efficacy study in patients with early diffuse disease. AB - OBJECTIVE: To investigate the efficacy of a treatment with low-dose intravenous cyclophosphamide (CYC) and low-dose prednisone in early diffuse cutaneous systemic sclerosis (dcSSc). METHODS: Patients with dcSSc and a disease duration <24 months consecutively admitted to a tertiary centre underwent a prospective 1 year study. They were treated with i.v. CYC 500 mg/pulses, 10 mg prednisone equivalent, and supportive therapy. Modified Rodnan skin score (mRss), Health Assessment Questionnaire-Disability Index (HAQ-DI), forced vital capacity (FVC), and diffusing lung capacity for CO (DLCO) were assessed as outcome measures. In addition, the nine Medsger severity scale scores were evaluated. RESULTS: mRss and DLCO significantly improved at both 6 (p = 0.002 and 0.012, respectively) and 12 months (p = 0.002 and 0.003, respectively). HAQ-DI showed a nearly significant reduction at 12 months (p = 0.06). Medsger's severity scores also improved for general condition (p = 0.001), peripheral vascular (p = 0.05), skin (p = 0.02), joint/tendon (p = 0.001), muscle (p = 0.05), and lung (p = 0.02). No treatment interruption was needed. CONCLUSIONS: This preliminary study suggests a role for low-dose i.v. CYC in the treatment of early dcSSc. Controlled studies are warranted. PMID- 16467040 TI - Association between the HLA-DRB1 gene and clinical features of systemic sclerosis in Korea. AB - OBJECTIVE: To determine whether HLA-DR alleles are associated with the development and clinical features of systemic sclerosis (SSc) in Koreans. METHODS: Seventy-nine patients (74 women and five men; 45 diffuse types and 34 limited types; mean age at diagnosis 43.9 years) fulfilling the American College of Rheumatology (ACR) classification criteria for SSc were enrolled. The controls were 144 healthy, disease-free Koreans. HLA-DRB1 genotypes were assessed by the polymerase chain reaction-sequence specific oligonucleotide probe (PCR-SSOP) method. RESULTS: The HLA-DRB1*15 allele was increased in anti-topoisomerase I autoantibody (anti-topo I)-positive SSc patients [p = 0.003, p corrected (p(corr)) = 0.039, odds ratio (OR) = 3.43, 95% confidence interval (CI) 1.45 8.13] compared with controls. The DRB1*11 allele was also observed more frequently in anti-topo I-positive SSc than in controls (13.3% vs. 4.2%) but not statistically significant (p = 0.053, p(corr) = 0.689). In patients with SSc, the DRB1*04 allele was associated with subcutaneous calcinosis (p = 0.048, OR = 4.56, 95% CI 1.07-19.37). Patients with overlap syndrome showed a negative association with the DRB1*04 allele (p = 0.036, OR = 0.26, 95% CI 0.08-0.91). CONCLUSION: The HLA-DRB1*15 allele was associated with the development of anti-topo I-positive SSc in Koreans. In addition, the DRB1*04 allele was associated with certain clinical features in SSc patients. PMID- 16467041 TI - Lack of association between TNFalpha gene polymorphism at position -308 and risk of acute rheumatic fever in Turkish patients. AB - OBJECTIVE: Acute rheumatic fever (ARF) is a multisystem inflammatory disease process that follows nasopharyngeal infection caused by group A streptococcus (GAS) (Streptococcus pyogenes). Recent studies have demonstrated that allelic variations at the tumour necrosis factor alpha (TNFalpha) locus are involved in the nature of rheumatic diseases such as juvenile idiopathic arthritis and rheumatic heart disease. Thus, TNFalpha polymorphisms at -308 in ARF patients might be useful in contributing to identification of the primary factors associated with pathogenesis of ARF. METHODS: We performed a case-control association study between the common G/A promoter polymorphism at position -308 in the TNFalpha gene and ARF in Turkish patients, investigating whether this locus acts as a risk factor or has a modifying effect. RESULTS AND CONCLUSION: Previous studies have reported that TNFalpha plays a major role in the pathogenesis of a number of autoimmune and inflammatory diseases. Moreover, significantly elevated TNFalpha levels were reported in patients with ARF. However, in our sample of patients with ARF (n = 66), no such association was found. No interactive effect was found between the TNFalpha polymorphism at position -308 and no association was detected with disease progression. These findings suggest that the role of TNFalpha in ARF may be in linkage disequilibrium with some other severity genes not yet genetically determined. PMID- 16467042 TI - Epidemiology and outcome of adult-onset Still's disease in Northern Norway. AB - BACKGROUND: Adult-onset Still's disease (AOSD) is considered a rare disease, but few data exist on the incidence and prevalence of AOSD. This study has analysed the epidemiology, disease presentation, and outcome of AOSD in a stable homogeneous population in Northern Norway. METHODS: A retrospective cohort study of all AOSD patients registered in 1990-2000 at the only hospital in the region with a Rheumatology Service. Clinical diagnosis and exclusion of patients were directed by the Yamaguchi criteria for AOSD. Demographic and clinical data at baseline were extracted from patient records and supplemented with data gathered at control visits. Data were analysed with nonparametric methods. RESULTS: AOSD was ascertained in 13 patients; the estimated annual AOSD incidence was 0.4/100,000 adults (95% CI 0.11-0.97), while point prevalence of AOSD increased from 3.4/100,000 (95% CI 0.8-9.4) in 1990 to 6.9/100,000 in 2000 (95% CI 2.7 14.2). Mean diagnostic delay was 5.2 months (range 0.5-18). Serum ferritin > 5 times the normal upper level had 63% diagnostic sensitivity. During 69 months' follow-up, one patient died, 6/13 patients achieved sustained remission, while six patients developed a chronic progressive (n = 3) or a relapsing/remitting disease course (n = 3). Four of these six patents had to enter social security programmes. CONCLUSION: The annual incidence of AOSD in Northern Norway is at least 0.4/100,000 adults. AOSD in this region is more prevalent than in France or Japan, affects more males, and approximates to the prevalence of juvenile Still's disease. Half of all patients have a monocyclic disease course, while mortality and invalidity occur in patients with chronic disease. PMID- 16467043 TI - Plasma levels of matrix metalloproteinase-9 in Henoch-Schonlein purpura. AB - OBJECTIVE: To investigate the potential role of matrix metalloproteinase-9 (MMP 9) in Henoch-Schonlein purpura (HSP), an acute type of systemic vasculitis in children. METHODS: In this study, 24 children with HSP and ten healthy children (HC) were enrolled from February 2003 to July 2004. Blood samples were obtained from all the children. The total levels of MMP-9 in the plasma were detected by enzyme-linked immunosorbent assay (ELISA). The second blood samples were obtained from eight of the 24 HSP patients in the convalescent phase. MMP-9 of circulating white blood cells was detected by immunocytochemistry. RESULTS: Plasma MMP-9 levels in the acute phase of HSP (249.75 ng/mL) were significantly higher than in HC (191.00 ng/mL) (p = 0.034). Immunocytochemistry showed that MMP-9 was positive in the circulating white blood cells. The MMP-9 levels in the convalescent phases were lower than in the acute phase in six cases, but increased in the other two cases, and one of these two cases had recurrence of purpuric rashes in the lower extremities for 3 months. CONCLUSION: MMP-9 plays an important role in the vascular destruction of HSP, and circulating white blood cells may be a source of the MMP-9 secreted into the circulation. PMID- 16467044 TI - Predictive factors for nephritis, relapse, and significant proteinuria in childhood Henoch-Schonlein purpura. AB - OBJECTIVE: To identify predictive factors for nephritis, relapse, and significant proteinuria in childhood Henoch-Schonlein purpura (HSP). METHODS: Two hundred and six consecutive patients with HSP (93 female, 113 male), followed up at a single centre between 1996 and 2001, were analysed retrospectively. They were regularly monitored for clinical and laboratory parameters for renal sequelae and relapse. RESULTS: Nephritis was seen in 78 patients (38%), relapse in 52 (25%), and significant proteinuria in 39 (19%). In univariate analysis, an older age at onset (>10 years), persistent purpura, severe bowel angina, and relapse were identified as factors associated with nephritis and significant proteinuria. Relapse-related factors were an older age, persistent purpura, severe bowel angina, and leucocytosis. Logistic regression analysis showed that nephritis was significantly associated with an older age, persistent purpura, and relapse, and significant proteinuria was closely related to severe bowel angina and relapse. CONCLUSION: We identified some predictors for nephritis, relapse, and significant proteinuria in childhood HSP, and close attention should be paid to those patients with the risk factors, such as an older age at onset, persistent purpura, severe bowel angina, and relapse. PMID- 16467045 TI - Evaluation of allantoin levels as a new marker of oxidative stress in Behcet's disease. AB - The increased production of reactive oxygen species (ROS) from activated neutrophils in Behcet's disease (BD) and recurrent aphthous stomatitis (RAS) may result in increased oxidative stress. Uric acid can react rapidly with neutrophil derived ROS to form allantoin. The purpose of the study was to evaluate the serum levels of allantoin as a new marker of oxidative stress in BD compared with malondialdehyde (MDA) levels as a well-known marker. Blood samples were obtained from 23 BD patients, 22 RAS patients as positive controls, and 21 healthy controls. When compared to the healthy controls, we found higher allantoin and MDA levels in the BD patients and higher MDA levels in the RAS patients. Serum ascorbic acid levels in the BD patients were significantly lower than in the controls. Increased allantoin and MDA levels suggest the possible involvement of free radicals in BD. As allantoin is only a product of uric acid oxidation by reactive oxygen and nitrogen species, it may also be used as a marker of oxidative stress in BD. PMID- 16467046 TI - Characteristics of vascular involvement in Behcet's disease. AB - OBJECTIVE: Behcet's disease (BD) is a multisystemic inflammatory disorder classified among the vasculitides, which can affect all types and sizes of blood vessels. Vascular involvement may be seen in 25-50% of BD patients. In this study, we examined the characteristics of vascular involvement in patients with BD. METHODS: One hundred and eighty patients with BD were included in the study. The diagnosis of vascular involvement was made on clinical signs, by Doppler ultrasonography and/or angiography using computed tomographic or magnetic resonance techniques where appropriate. Detailed clinical characteristics were recorded for each patient. RESULTS: Seventy-one patients (39.4%) had vascular involvement. In patients with vascular lesions, the frequency of male sex was significantly higher than in patients without vascular lesions (89.8% vs. 63.3%, respectively; p < 0.001). Of 71 BD patients with vascular involvement, 68 had venous lesions (95.8%). Three patients had arterial lesions without venous thrombosis. Eleven patients had arterial involvement with venous thrombosis. The most frequent type of vascular involvement was deep venous thrombosis in the lower extremities (n = 56, 78.9%). There was a significant association between deep venous thrombosis and superficial thrombophlebitis (r = 0.325, p < 0.01). Twenty-four patients (33.8%) had vena cava thrombosis and two had vena hepatica thrombosis. In patients with vascular involvement, the frequency of erythema nodosum was significantly higher (p = 0.001) and the frequency of ocular involvement was significantly lower (p < 0.05) than in patients without vascular involvement. CONCLUSION: Our study illustrates the frequency and significance of vascular involvement in BD. PMID- 16467048 TI - Progression of imaging in pancreatitis panniculitis polyarthritis (PPP) syndrome. AB - Lobular panniculitis, together with a polyarthritis may complicate pancreatic disease. Abdominal symptoms are frequently absent and mis-diagnosis of the joint skin complex can lead to inappropriate treatment in a condition with an already high mortality. We report a case of the pancreatitis panniculitis polyarthritis (PPP) syndrome, complicated by metastatic fat necrosis, with bone marrow involvement and characteristic magnetic resonance imaging (MRI) and describe the clinical characteristics, therapy and outcome of patients affected by the syndrome. PMID- 16467047 TI - A case of IgG4-related autoimmune disease with multiple organ involvement. AB - A 52-year-old male was admitted with autoimmune pancreatitis (AIP), showing mononuclear cell infiltration in both the pancreas and salivary glands with both normal sialography and anti-SS-A/SS-B antibodies. Although the AIP improved with glucocorticoid treatment, subsequent abdominal computed tomography (CT) revealed a nodular shadow in the bilateral kidneys, which was confirmed as interstitial nephritis by renal biopsy. The patient's serum immunoglobulin G4 (IgG4) level was 10 times higher than the upper limit of the normal range. IgG4-positive mononuclear cell infiltration was detected in the salivary gland, pancreas, and kidney. A new entity proposed as 'IgG4-related autoimmune disease' was considered. PMID- 16467049 TI - HLA class II allele polymorphism in Hungarian patients with primary Sjogren's syndrome. PMID- 16467050 TI - Fine-needle aspiration cytology, salivary gland ultrasonography, and sialography in the evaluation of primary Sjogren's syndrome. PMID- 16467051 TI - Thrombocytopaenia in juvenile dermatomyositis. PMID- 16467052 TI - A case of autoimmune polyglandular syndrome type III with a slowly progressive form of type-1 diabetes mellitus that manifested in the course of autoimmune diseases. PMID- 16467053 TI - The epidemiology of cerebral palsy: incidence, impairments and risk factors. AB - PURPOSE: Describing the epidemiology of cerebral palsy (CP), its impairments and risk factors. METHOD: Literature review 1965-2004. Search terms: Cerebral palsy, incidence, prevalence, impairments, risk factors. RESULTS: In the last 40 years the prevalence of CP has risen to well above 2.0 per 1000 life births. In this time span the proportion of low-birthweight infants rose, the proportion of diplegia decreased, while the proportion of hemiplegia increased. CP is more prevalent in more deprived socio-economic populations. The majority of people with CP have the spastic syndrome of which the diplegic group is the smallest. Dependent on the subgroup of CP, 25-80% have additional impairments. A large proportion has some kind of cognitive impairment; the prevalence varies with the type of CP and especially increases when epilepsy is present. Epilepsy is present in 20-40%; it is most common among the hemi- and tetraplegics. Sensibility of the hands is impaired in about half. Chronic pain is reported by more than a quarter of the adults. Up to 80% have at least some impairment of speech. Low visual acuity is reported in almost three-quarters of all children. Half of all children have gastrointestinal and feeding problems. Stunted growth occurs in a quarter, while under- or overweight problems are present in half of the children. Almost 70% of people with spastic CP have abnormal brain CT findings; abnormal cranial ultrasounds is most strongly associated with hemiplegia, normal cranial ultrasounds with diplegia. The most important risk factors for CP are low birthweight, intrauterine infections and multiple gestation. PMID- 16467054 TI - The Participation Scale: measuring a key concept in public health. AB - PURPOSE: To develop a scale to measure (social) participation for use in rehabilitation, stigma reduction and social integration programmes. METHOD: A scale development study was carried out in Nepal, India and Brazil using standard methods. The instrument was to be based on the Participation domains of the International Classification of Functioning, Disability and Health (ICF), be cross-cultural in nature and assess client-perceived participation. Respondents rated their participation in comparison with a "peer", defined as "someone similar to the respondent in all respects except for the disease or disability". RESULTS: An 18-item instrument was developed in seven languages. Crohnbach's alpha was 0.92, intra-tester stability 0.83 and inter-tester reliability 0.80. Discrimination between controls and clients was good at a Participation Score threshold of 12. Responsiveness after a "life change" was according to expectation. CONCLUSIONS: The Participation Scale is reliable and valid to measure client-perceived participation in people affected by leprosy or disability. It is expected to be valid in other (stigmatised) conditions also, but this needs confirmation. The scale allows collection of participation data and impact assessment of interventions to improve social participation. Such data may be compared between clients, interventions and programmes. The scale is suitable for use in institutions, but also at the peripheral level. PMID- 16467055 TI - What determines the possession of assistive devices among patients with rheumatic diseases? The influence of the country-related health care system. AB - PURPOSE: To identify the determinants of the possession of assistive devices among patients with various rheumatic conditions. In order to determine the influence of the country-related health care system, patients from two different countries were studied. METHOD: Patients with rheumatoid arthritis (RA) or psoriatic arthritis (PsA) were selected from rheumatology outpatient clinics in two adjacent regions in The Netherlands and Germany. A total of 142 patients completed a self-administered questionnaire. Information on the possession of assistive devices and data on socio-demographics, clinical status and health status were obtained. Logistic regression analyses were used to identify determinants of the possession of assistive devices. RESULTS: The majority (78%) of the patients possessed at least one or more assistive devices. Obviously, functional status was the most important determinant, followed by the country where the patient resided. More assistive devices were found in increasingly disabled patients as well as in patients living in The Netherlands. CONCLUSION: Functional status and the patient's country are the most important determinants of the possession of assistive devices among patients with rheumatic conditions. We hypothesize that the most likely explanation for the differences in possession rates between countries are differences in societal systems for the prescription and reimbursement of assistive devices. PMID- 16467056 TI - Comparison between performance with a pushrim-activated power-assisted wheelchair and a manual wheelchair on the Wheelchair Skills Test. AB - PURPOSE: To test the hypothesis that people using a pushrim-activated power assisted wheelchair (PAPAW) can accomplish a wider range of wheelchair skills than when using a manual wheelchair (MWC). METHODS: We studied 30 able-bodied participants, using within-participant comparisons. Participants used a manual wheelchair equipped with both PAPAW and regular MWC rear wheels, and rear anti tip devices (Arc-RADs) that permitted wheelie-like function. We trained participants to perform the wheelchair skills of the Wheelchair Skills Training Program (WSTP, Version 2.4). From the Wheelchair Skills Test (WST, Version 2.4), we calculated pass-fail success rates for the 50 individual skills and a total percentage WST score. RESULTS: The mean (+/-SD) total WST scores were 89.3 (+/ 7.0)% for the PAPAW and 88.8 (+/-8.4)% for the MWC, with a mean difference of 0.6 (+/-5.6)% (p = 0.59). Qualitative observations suggested that skills requiring a higher force on the pushrim (e.g., incline ascent) were performed more easily with the PAPAW, whereas skills requiring greater control of the wheelchair (e.g., wheelie-dependent skills) were performed more easily with the MWC. CONCLUSION: Overall wheelchair skill performance with the PAPAW is not superior to that when using the MWC. The PAPAW may be helpful for specific skills that require more wheel torque, but the additional torque appears to be disadvantageous when performing skills that require greater control. PMID- 16467058 TI - Application of facial electromyography in computer mouse access for people with disabilities. AB - PURPOSE: This study develops a newly facial EMG human-computer interface for people with disabilities for controllng the movement of the cursor on a computer screen. METHOD: We access the computer cursor according to different facial muscle activity patterns. In order to exactly detect the muscle activity threshold, this study adopts continuous wavelet transformation to estimate the single motor unit action potentials dynamically. RESULT: The experiment indicates that the accuracy of using the facial mouse is greater than 80%, and this result indicates the feasibility of the proposed system. Moreover, the subject can improve performance of manipulation by repeated training. CONCLUSION: Compared with previous works, the proposed system achieves complete cursor function and provides an inexpensive solution. Although there are still some drawbacks in the facial EMG-based human-computer interface, the facial mouse can provide an alternative among other expensive and complicated assistive technologies. PMID- 16467057 TI - Predictors of long-term participation after stroke. AB - PURPOSE: (1) To explore factors that predict long-term participation after stroke (2-4 years after discharge from rehabilitation), and (2) to determine factors that predict both short- and long-term participation. METHODS: Biopsychosocial data of people who had had a stroke were measured at discharge from an intensive rehabilitation unit using valid instruments. Six months later (n=102) as well as 2-4 years later (n=66), social participation of the survivors was measured in their living environments. Participation was estimated with the Assessment of Life Habits (LIFE-H), which includes 12 categories of daily activities and social roles. RESULTS: From multivariate regression analyses, the best predictors of long-term participation after stroke appear to be age, comorbidity, motor coordination, upper extremity ability and affect. Age, comorbidity, affect and lower extremity coordination are the best predictors of participation after stroke at both measurement times. CONCLUSIONS: With the exception of age, these factors may be positively modified and thus warrant special attention in rehabilitation interventions. PMID- 16467059 TI - A review of the aetiological factors and results of trans-ankle (Syme) disarticulations. AB - INTRODUCTION: Ever since James Syme described his amputation at the ankle joint, most authors have agreed that Syme's amputation should not be considered for patients with diabetic vascular disease (DVD). The aim of the study was to record our experience with Syme's amputations and to search for any predictive criteria that may enhance successful results. PATIENTS AND METHODS: Seventy patients underwent a Syme amputation between the years 1980 and 2000 in our department, 51 owing to DVD. The other 19 patients had their amputations because of other pathologies. RESULTS: The 19 patients of the non-DVD group showed a success rate of 94.7% (18/19). The overall success rate of the DVD patients was 49% (25/51). When we analysed the DVD group by age, the cut-off point was 65 years old. Patients aged 65 or less had a success rate of 68.2% (15/22), and over the age of 65 years the success rate was only 30.8% (8/26). CONCLUSIONS: Patients with DVD should be considered good candidates for a Syme amputation if they are younger than 65 years old. PMID- 16467060 TI - Cemented versus uncemented hip replacement. PMID- 16467061 TI - Disabilities, physicians and the Millennium Development Goals. PMID- 16467062 TI - Sequestered hypoplastic pulmonary lobe supplied by the circumflex coronary artery in a patient with coronary artery disease: a case report. AB - Bronchopulmonary sequestrations are usually supplied by single or multiple branches from the descending aorta. We report on a rare form of sequestration of a hypoplastic mid-pulmonary lobe with arterial supply from the circumflex coronary artery coincidentally discovered during coronary angiography in a female patient with coronary artery disease and a history of recurrent hemoptysis. The patient underwent myocardial revascularization and resection of the sequestered lobe in a single-stage approach. PMID- 16467063 TI - Prospective, randomized study on the use of the cardica PAS-port aortic connector system in off-pump coronary artery bypass surgery. AB - BACKGROUND: The use of aortic connector devices for proximal vein graft anastomosis has been shown to be associated with a relevant rate of early graft complications. Cardica PAS-Port is a new aortic connector whose preliminary clinical results seem promising. The safety and efficacy of this aortic connector device have been evaluated in this prospective, randomized study. MATERIAL AND METHODS: Twenty-four patients were randomized to receive proximal aorta-vein graft anastomosis with either the Cardica PAS-port aortic connector or by the hand-sewn technique. Twenty-three patients underwent multidetector computed tomographic scan (MDCT) of the chest 6 months after surgery to evaluate graft patency. RESULTS: All aortic connector devices (18) were successfully deployed and 31 proximal anastomoses were performed by the hand-sewn technique. MDCT showed that 6-month freedom from vein graft complication was 22.2% in the PAS Port group and 58.1% in the hand-sewn group (P = .04). Four vein grafts (22.2%) anastomosed with the PAS-Port and 2 hand-sewn vein grafts (6.5%) were occluded (P = .10). The use of the PAS-Port aortic connector was also predictive of any vein graft complication when adjusted for vein graft flow (P = .01; OR 8.64, 95% CI 1.66-45.00) and for peripheral resistance units (P = .02; OR 6.14, 95% CI 1.33 28.43). CONCLUSIONS: The results of this prematurely stopped, prospective, randomized study suggest that the use of PAS-Port aortic connector device is associated with a higher rate of early vein graft complications than the hand sewn technique. PMID- 16467064 TI - Sixteen-channel multidetector row computed tomography versus coronary angiography in a surgical view. AB - BACKGROUND: Invasive coronary angiography (ICA) is the gold standard for the diagnosis of coronary artery disease and also for imaging procedures for preoperative planning of coronary artery bypass grafting (CABG). Sixteen multidetector row computed tomography (MDCT) represents an alternative depiction of coronary vessels. METHODS: Preoperative exams included ICA and MDCT in 50 patients. Two blinded surgical readers independently investigated both diagnostic modalities regarding location, severity, and morphology of the stenoses. The right coronary artery, left anterior descending branch, and circumflex branch- each divided in 3 sections--and the left main artery with a diameter (3) 1.5 mm were rated in both procedures, and the percentage of complete evaluations by MDCT was assessed. RESULTS: Heart rate was 72 +/- 8 bpm. Forty-six percent of patients received a complete MDCT evaluation, and 54% received an incomplete MDCT evaluation. In 62% of these incompletely examined patients, 1 branch was not completely analyzable, in 31% 2 branches; and in 7% all 3 branches. In total, 9% of all segments were incompletely assessed. Investigators detected coronary stenoses in complete evaluations with a sensitivity of 94% and a specificity of 95%. Positive predictive value was 87% and negative predictive value was 98%. Plaque classification in soft and hard plaques was possible. CONCLUSION: Sixteen MDCT is not a viable alternative diagnostic tool at present. However, although the percentage of incomplete evaluated patients is more then 50%, only 9% of all segments were incompletely assessable. If this technology can be further improved, especially its software, it will become a valid diagnostic tool for coronary artery disease. PMID- 16467065 TI - Direct left ventricle to great cardiac vein retroperfusion: a novel alternative to myocardial revascularization. AB - BACKGROUND: As the number of patients with diffuse coronary artery disease continues to grow, there is renewed interest in alternative methods of perfusing the ischemic myocardium. We tested the feasibility of myocardial retroperfusion via a direct left ventricle-to-great cardiac vein (LV-GCV) conduit to support regional contractility in this setting. METHODS: LV-GCV flow was established using an extracorporeal circuit in 5 dogs. Left ventricle (LV) pressure, aortic pressure, regional myocardial segment length, and circuit blood flow were measured prior to left anterior descending coronary artery (LAD) ligation, following LAD ligation, and after LV-GCV circuit placement. To eliminate backward flow during diastole, an in-line flow regulator was placed. Regional myocardial function was quantified by pressure-segment length loop area divided by end diastolic segment length (PSLA/EDSL). RESULTS: LAD ligation reduced PSLA/EDSL from 10.0 +/- 1.2 mm Hg mm to 1.6 +/- 0.3 mm Hg mm (P < .05). With LV-GCV retroperfusion, mean peak systolic flow was +152 +/- 14 mL/min, mean peak diastolic flow was -39 +/- 11 mL/min, and net mean flow was +36 +/- 13 mL/min. Regional function recovered to approximately 39% of baseline (3.9 +/- 0.4 mm Hg mm, P < .05). Upon elimination of backflow, mean flow increased to +41 +/- 12 mL/min and regional function recovered even further to approximately 47% of baseline (4.6 +/- 0.7 mm Hg mm, P < .05). CONCLUSIONS: A LV-GCV circuit can significantly restore regional function to the acutely ischemic myocardium. An inline valve that eliminates backward diastolic flow improves regional function even further. This approach may provide an effective therapy for diffuse coronary disease not amenable to traditional revascularization strategies. PMID- 16467066 TI - Multiple left ventricular myxomas arising from the interventricular septum and ventricular trabeculae: a case report. AB - Myxoma is the most common tumor of the heart, with an estimated incidence of 0.5 to 1 per million per year. The majority of these tumors arise from the left atrium, and 20% arise from the right atrium with the remaining 5% arising in either the right or left ventricle. We present a rare case of multiple myxomas of the left ventricle arising from the interventricular septum and nearby ventricular trabeculae that clinically presented with an occlusion of the left axillary artery. The patient was successfully treated using cardiopulmonary bypass, during which the tumor was completely removed using a transventricular approach. PMID- 16467067 TI - Combined physiological effects of bronchodilators and hyperoxia on exertional dyspnoea in normoxic COPD. AB - BACKGROUND: Studies examining the physiological interactions of oxygen (O(2)) and bronchodilators (BD) during exercise in chronic obstructive pulmonary disease (COPD) should provide new insights into mechanisms of exercise intolerance. We examined the effects of O(2) and BD, alone and in combination, on dyspnoea, ventilation (e), breathing pattern, operating lung volumes, and exercise endurance. METHODS: In a randomised, double blind, crossover study, 16 patients with COPD (mean (SE) FEV(1) 43(3)% predicted) performed pulmonary function tests and an incremental exercise test, then completed four visits in which they received either nebulised BD (ipratropium 0.5 mg + salbutamol 2.5 mg) or placebo (PL) with either 50% O(2) or room air (RA). After 90-105 minutes the patients performed pulmonary function tests, then breathed RA or O(2) during symptom limited constant load exercise at 75% peak work rate. RESULTS: With BD the mean (SE) increase in inspiratory capacity (IC) was 0.3 (0.1) l (p<0.05) at rest and during exercise, permitting greater tidal volume (Vt) expansion during exercise and a greater peak e. With O(2), e decreased during exercise as a result of decreased breathing frequency (F), with no significant change in IC. During exercise with BD+O(2), IC and Vt increased, F decreased, and e did not change. Dyspnoea decreased with all interventions at a standardised time during exercise compared with PL+RA (p<0.05). Endurance time was significantly (p<0.05) greater with BD+O(2) (10.4 (1.6) min) than with O(2) (8.5 (1.4) min), BD (7.1 (1.3) min) and PL+RA (5.4 (0.9) min). CONCLUSION: By combining the benefits of BD (reduced hyperinflation) and O(2) (reduced ventilatory drive), additive effects on exercise endurance were observed in patients with normoxic COPD. PMID- 16467068 TI - Performance of mechanical ventilators at the patient's home: a multicentre quality control study. AB - BACKGROUND: Quality control procedures vary considerably among the providers of equipment for home mechanical ventilation (HMV). METHODS: A multicentre quality control survey of HMV was performed at the home of 300 patients included in the HMV programmes of four hospitals in Barcelona. It consisted of three steps: (1) the prescribed ventilation settings, the actual settings in the ventilator control panel, and the actual performance of the ventilator measured at home were compared; (2) the different ventilator alarms were tested; and (3) the effect of differences between the prescribed settings and the actual performance of the ventilator on non-programmed readmissions of the patient was determined. RESULTS: Considerable differences were found between actual, set, and prescribed values of ventilator variables; these differences were similar in volume and pressure preset ventilators. The percentage of patients with a discrepancy between the prescribed and actual measured main ventilator variable (minute ventilation or inspiratory pressure) of more than 20% and 30% was 13% and 4%, respectively. The number of ventilators with built in alarms for power off, disconnection, or obstruction was 225, 280 and 157, respectively. These alarms did not work in two (0.9%), 52 (18.6%) and eight (5.1%) ventilators, respectively. The number of non programmed hospital readmissions in the year before the study did not correlate with the index of ventilator error. CONCLUSIONS: This study illustrates the current limitations of the quality control of HMV and suggests that improvements should be made to ensure adequate ventilator settings and correct ventilator performance and ventilator alarm operation. PMID- 16467069 TI - Increased risk of chronic obstructive pulmonary diseases in coke oven workers: interaction between occupational exposure and smoking. AB - BACKGROUND: Coke oven workers are regularly exposed to coke oven emissions (COE) and may be at risk of developing lung diseases. There is limited evidence for the link between exposure to COE and chronic obstructive pulmonary diseases (COPD). The aim of this study was to explore the dose-response relationship between COE exposure and COPD and to assess the interaction with cigarette smoking. METHODS: Seven hundred and twelve coke oven workers and 211 controls were investigated in southern China. Benzene soluble fraction (BSF) concentrations as a surrogate of COE were measured in representative personal samples and the individual cumulative COE exposure level was quantitatively estimated. Detailed information on smoking habits and respiratory symptoms was collected and spirometric tests were performed. RESULTS: The mean BSF levels at the top of two coking plants were 743.8 and 190.5 microg/m3, respectively, which exceed the OSHA standard (150 microg/m3). After adjusting for cigarette smoking and other risk factors, there was a significant dose-dependent reduction in lung function and increased risks of chronic cough/phlegm and COPD in coke oven workers. The odds ratio for COPD was 5.80 (95% confidence interval 3.13 to 10.76) for high level cumulative COE exposure (> or =1714.0 microg/m3-years) compared with controls. The interaction between COE exposure and smoking in COPD was significant. The risk of COPD in those with the highest cumulative exposure to COE and cigarette smoking was 58 fold compared with non-smokers not exposed to COE. CONCLUSION: Long term exposure to COE increases the risk of an interaction between COPD and cigarette smoking. PMID- 16467070 TI - Significance of non-calcified pulmonary nodules in patients with extrapulmonary cancers. AB - BACKGROUND: This study sought to determine the rate and patterns of malignancy in patients with extrapulmonary cancers and non-calcified pulmonary nodules, and to develop a statistical model to guide clinicians regarding choice of patients for diagnostic biopsy. METHOD: The medical records of 151 patients evaluated at the Memorial Sloan-Kettering Cancer Center between January 1999 and December 2001 for non-calcified pulmonary nodules were reviewed. Nodules were considered malignant based on the results of a diagnostic biopsy, and were considered benign if their appearance remained stable 2 years after the initial study, if they resolved, or if a biopsy showed a non-malignant condition. RESULTS: Sixty four of 151 patients (42%) were diagnosed with malignant nodules; 32 had newly diagnosed lung cancers, 28 had metastatic spread of their primary cancers, and four had lesions that were either new cancers or of undetermined aetiology. On univariate analysis the likelihood of malignancy increased with nodule size, tobacco exposure, and the finding of a solitary nodule. On multivariable analysis only nodule size and tobacco exposure were predictive of malignancy. The model had good predictive accuracy (area under the curve 0.751) but had insufficient discrimination for use as a clinical tool to determine which patients should undergo diagnostic biopsy. CONCLUSION: Nearly half the non-calcified pulmonary nodules identified in this series were malignant. Lung cancer was more common than metastatic disease. These findings support the need for close interval follow up and a low threshold for diagnostic biopsy in patients with extrapulmonary cancers and non-calcified pulmonary nodules. In smokers, such lesions should raise concern for lung cancer. PMID- 16467071 TI - Relation of interlobar collaterals to radiological heterogeneity in severe emphysema. AB - BACKGROUND: A study was undertaken to assess the prevalence of interlobar collateral ventilation in patients with severe emphysema to identify factors that may help to predict patients with significant collateral ventilation. METHODS: Between April 2002 and August 2003, ex vivo assessment of the lungs 17 consecutive patients with smoking related severe emphysema was performed. To assess collateral flow, all lobes of explanted specimens were selectively intubated using a wedged cuffed microlaryngeal intubation tube and then manually ventilated using a bagging circuit. Interlobar collateral ventilation was defined as the ability to easily inflate a non-intubated lobe at physiological pressures. Pre-transplant demographic characteristics, physiological data, radiological results, and explant histology were assessed for retrospective relationships with the degree of interlobar collateral ventilation in the explanted lung. RESULTS: A total of 23 lungs were evaluated, 15 of which (66%) had significant collateral interlobar airflow. There were no significant differences in any demographic, physiological, or pathological variables between patients with collateral ventilation and those with no collateral ventilation. However, there was a significant relationship between the presence of interlobar collateral ventilation and radiological scores (p<0.05). CONCLUSIONS: Interlobar collateral ventilation occurs to a much greater extent in patients with radiologically homogeneous emphysema than in those with heterogeneous emphysema. Heterogeneity of emphysema may predict patients with a significantly reduced risk of interlobar collateral ventilation. PMID- 16467072 TI - Continuous positive airway pressure reduces daytime sleepiness in mild to moderate obstructive sleep apnoea: a meta-analysis. AB - BACKGROUND: Obstructive sleep apnoea syndrome (OSAS) affects an estimated 2-4% of the middle aged population. Meta-analyses of randomised controlled trials have shown that the severe presentation of the syndrome (apnoea hypopnoea index (AHI) >30/hour) is effectively treated with continuous positive airway pressure (CPAP). Until recently there have been insufficient data to determine whether CPAP improves sleepiness in the larger subgroup with mild to moderate OSAS (AHI 5 30/hour). METHODS: A systematic search of Medline and a hand search identified seven randomised controlled trials where CPAP was compared with either a placebo or with conservative management in the treatment of mild to moderate OSAS (AHI 5 30/hour). All trials used the Epworth Sleepiness Scale (ESS), four used the Multiple Sleep Latency Test (MSLT), and three used the Maintenance of Wakefulness Test (MWT) to measure sleepiness. RESULTS: Meta-analyses indicated that CPAP significantly reduced subjective daytime sleepiness (ESS) by 1.2 points (95% CI 0.5 to 1.9, p = 0.001), improved objective daytime wakefulness (MWT) by 2.1 minutes (95% CI 0.5 to 3.7, p = 0.011), but did not affect objective daytime sleepiness (MSLT, mean benefit -0.2 minutes, 95% CI -1.0 to 0.6, p = 0.6). The two significant effects were small (effect size <0.30). CONCLUSIONS: CPAP elicits small improvements in subjective sleepiness and objective wakefulness in people with mild to moderate OSAS. However, the effects on sleepiness are of limited clinical significance. PMID- 16467073 TI - Functional variants of antioxidant genes in smokers with COPD and in those with normal lung function. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is predominantly the consequence of chronic smoking exposure, but its development may be influenced by genetic variants that affect lung remodelling, inflammation, and defence from oxidant stress. A study was undertaken to determine whether genetic variants within genes encoding the antioxidant enzymes superoxide dismutase (SOD) and catalase may be associated with the development of impaired lung function. METHODS: In a case-control study, the allele and genotype frequencies of functional polymorphisms from SOD1 (CuZnSOD), SOD2 (MnSOD), SOD3 (extracellular SOD), and catalase (CAT) were compared in chronic smokers with normal lung function (resistant smokers) and in those with COPD. RESULTS: Significantly higher frequencies of the G allele and CG/GG genotype of the 213 SOD3 polymorphism were found in resistant smokers (odds ratios (ORs) 4.3 (95% CI 1.5 to 13.3) and 4.2, 95% CI 1.4 to 13.3), Bonferroni corrected p = 0.02 and p = 0.02, respectively) than in those with COPD. There were no differences between the COPD and resistant smokers for the SOD1, SOD2, or CAT polymorphisms tested. CONCLUSIONS: The 213Gly variant of the SOD3 gene may, through antioxidant or anti inflammatory effects, confer a degree of resistance in some smokers to the development of COPD. PMID- 16467074 TI - Longitudinal pulmonary function of childhood bronchiectasis and comparison with cystic fibrosis. AB - BACKGROUND: Little has been published on the progression of non-cystic fibrosis bronchiectasis (BX), especially in childhood. Data are needed for prognosis and evaluation of the effectiveness of treatments. A study was undertaken to evaluate the change in lung function over time in children with BX, and to consider covariates and compare them with the local cystic fibrosis (CF) population. METHODS: Children with BX or CF and > or =3 calendar years of lung function data were identified from hospital clinics. Diagnosis was made by high resolution CT scans, sweat tests, and genetic studies. Lung function performed on a single plethysmograph between 6 and 15 years of age and > or =6 weeks after diagnosis was analysed longitudinally (linear mixed model). The impact of reference equation and "best annual" versus "all data" approaches were evaluated. RESULTS: There were 44 children in each of the BX and CF groups with an overall mean 5.7 calendar years follow up data. The estimated forced expiratory volume in 1 second (FEV(1)) in the BX group had an intercept of 68% predicted (Polgar) at 10 years of age which fell at a rate of 1.9% per annum using "best annual" data compared with 63% and 0.9% using "all data". Those with post-infectious BX or chronic Haemophilus influenzae infection had more severe disease. In CF the FEV(1) ("best annual") intercept was 85% predicted with a slope of -2.9% per annum. The choice of reference equation affected the magnitude of the result but not the conclusions. CONCLUSION: Children with BX have significant airway obstruction which deteriorates over time, regardless of analysis strategy or reference. Effective interventions are needed to prevent significant morbidity and adult mortality. PMID- 16467076 TI - Well-defined melanoma antigens as progression markers for melanoma: insights into differential expression and host response based on stage. PMID- 16467075 TI - Dietary antioxidants and asthma in adults. AB - BACKGROUND: Several antioxidant nutrients have been reported to be inversely associated with asthma. A study was undertaken to assess the independent associations of these nutrients with asthma in adults. METHODS: A nested case control study was performed in 515 adults with physician diagnosed asthma and 515 matched controls using dietary data obtained from 7 day food diaries. The main outcome measures were physician diagnosed asthma and current symptomatic asthma (diagnosed asthma and self-reported wheeze within the previous 12 months). RESULTS: Cases were similar to controls in age, sex, social class, and daily energy intake but had a lower median intake of fruit (132.1 v 149.1 g/day, p< or =0.05). 51.5% of the population reported zero consumption of citrus fruit; relative to these individuals, people who consumed >46.3 g/day had a reduced risk of diagnosed and symptomatic asthma (OR adjusted for potential confounders 0.59 (95% CI 0.43 to 0.82) and 0.51 (95% CI 0.33 to 0.79), respectively). In nutrient analysis, dietary vitamin C and manganese were inversely and independently associated with symptomatic asthma (adjusted OR per quintile increase 0.88 (95% CI 0.77 to 1.00) for vitamin C and 0.85 (95% CI 0.74 to 0.98) for manganese), but only manganese was independently associated with diagnosed asthma (OR 0.86 (95% CI 0.77 to 0.95)). Adjusted plasma levels of vitamin C were significantly lower in symptomatic cases than in controls (54.3 v 58.2 micromol/l, p = 0.003). CONCLUSIONS: Symptomatic asthma in adults is associated with a low dietary intake of fruit, the antioxidant nutrients vitamin C and manganese, and low plasma vitamin C levels. These findings suggest that diet may be a potentially modifiable risk factor for the development of asthma. PMID- 16467077 TI - Handicapping the race to develop inhibitors of the phosphoinositide 3 kinase/Akt/mammalian target of rapamycin pathway. AB - The phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) pathway controls many cellular processes that are important for the formation and progression of cancer, including apoptosis, transcription, translation, metabolism, angiogenesis, and cell cycle progression. Genetic alterations and biochemical activation of the pathway are frequent events in preneoplastic lesions and advanced cancers and often portend a poor prognosis. Thus, inhibition of the PI3K/Akt/mTOR pathway is an attractive concept for cancer prevention and/or therapy. Inhibitors of individual components, such as PI3K, PDK-1, Akt, and mTOR, are being developed at a rapid pace and have promise for improving the care of cancer patients. Here, we review the published data on inhibitors of the pathway and discuss relevant issues, such as the complex regulation of the pathway, the design of clinical trials, and the likelihood of finding a therapeutic index when targeting such a critical signaling pathway. PMID- 16467078 TI - Expression profiling of mucinous tumors of the ovary identifies genes of clinicopathologic importance. AB - PURPOSE: To elucidate the molecular mechanisms contributing to the unique clinicopathologic characteristics of mucinous ovarian carcinoma, global gene expression profiling of mucinous ovarian tumors was carried out. EXPERIMENTAL DESIGN: Gene expression profiling was completed for 25 microdissected mucinous tumors [6 cystadenomas, 10 low malignant potential (LMP) tumors, and 9 adenocarcinomas] using Affymetrix U133 Plus 2.0 oligonucleotide microarrays. Hierarchical clustering and binary tree prediction analysis were used to determine the relationships among mucinous specimens and a series of previously profiled microdissected serous tumors and normal ovarian surface epithelium. PathwayAssist software was used to identify putative signaling pathways involved in the development of mucinous LMP tumors and adenocarcinomas. RESULTS: Comparison of the gene profiles between mucinous tumors and normal ovarian epithelial cells identified 1,599, 2,916, and 1,765 differentially expressed in genes in the cystadenomas, LMP tumors, and adenocarcinomas, respectively. Hierarchical clustering showed that mucinous and serous LMP tumors are distinct. In addition, there was a close association of mucinous LMP tumors and adenocarcinomas with serous adenocarcinomas. Binary tree prediction revealed increased heterogeneity among mucinous tumors compared with their serous counterparts. Furthermore, the cystadenomas coexpressed a subset of genes that were differentially regulated in LMP and adenocarcinoma specimens compared with normal ovarian surface epithelium. PathwayAssist highlighted pathways with expression of genes involved in drug resistance in both LMP and adenocarcinoma samples. In addition, genes involved in cytoskeletal regulation were specifically up-regulated in the mucinous adenocarcinomas. CONCLUSIONS: These data provide a useful basis for understanding the molecular events leading to the development and progression of mucinous ovarian cancer. PMID- 16467079 TI - Gene expression differences associated with human papillomavirus status in head and neck squamous cell carcinoma. AB - Human papillomavirus (HPV) is associated with a subset of head and neck squamous cell carcinoma (HNSCC). Between 15% and 35% of HNSCCs harbor HPV DNA. Demographic and exposure differences between HPV-positive (HPV+) and negative (HPV-) HNSCCs suggest that HPV+ tumors may constitute a subclass with different biology, whereas clinical differences have also been observed. Gene expression profiles of HPV+ and HPV- tumors were compared with further exploration of the biological effect of HPV in HNSCC. Thirty-six HNSCC tumors were analyzed using Affymetrix Human 133U Plus 2.0 GeneChip and for HPV by PCR and real-time PCR. Eight of 36 (22%) tumors were positive for HPV subtype 16. Statistical analysis using Significance Analysis of Microarrays based on HPV status as a supervising variable resulted in a list of 91 genes that were differentially expressed with statistical significance. Results for a subset of these genes were verified by real-time PCR. Genes highly expressed in HPV+ samples included cell cycle regulators (p16(INK4A), p18, and CDC7) and transcription factors (TAF7L, RFC4, RPA2, and TFDP2). The microarray data were also investigated by mapping genes by chromosomal location (DIGMAP). A large number of genes on chromosome 3q24-qter had high levels of expression in HPV+ tumors. Further investigation of differentially expressed genes may reveal the unique pathways in HPV+ tumors that may explain the different natural history and biological properties of these tumors. These properties may be exploited as a target of novel therapeutic agents in HNSCC treatment. PMID- 16467080 TI - Molecular context of the EGFR mutations: evidence for the activation of mTOR/S6K signaling. AB - PURPOSE: Activating somatic mutations in the epidermal growth factor receptor (EGFR) gene are present in a small subset of lung adenocarcinomas. These mutations cluster in specific regions and confer sensitivity to inhibitors of the tyrosine kinase activity of EGFR. To further determine the genetic and molecular characteristics of tumors carrying EGFR gene mutations, we investigated the EGFR gene status in lung adenocarcinomas and evaluated its association with specific characteristics of the patients and tumors, such as mutations at KRAS and p53, EGFR and ErbB2 gene amplification, levels of EGFR and HER2 proteins, and levels of downstream effectors of EGFR, such as phospho-extracellular signal-regulated kinase and phospho-S6 proteins. EXPERIMENTAL DESIGN: The mutational status of EGFR was determined by direct sequencing in 86 primary lung adenocarcinomas and 12 lung cancer cell lines, and was correlated with a number of variables relating to the tumor and patient. A tissue microarray containing 37 lung tumors was constructed to determine, by fluorescence in situ hybridization analysis, the number of copies of EGFR and ErbB2 genes and, by immunohistochemistry, the levels of EGFR, HER2, phospho-ERK, and phospho-S6 proteins. RESULTS: EGFR gene mutations were identified in 13% of the primary tumors. The type and clustering of the mutations were identical to those previously reported. Amplification of the EGFR occurred in 14% of the tumors and could arise in tumors with EGFR mutations. Interestingly, mTOR activation, as measured indirectly by augmented levels of phospho-S6 protein, was more frequent in tumors with gene alterations in either EGFR or KRAS (P = 0.00005; Fisher's exact test) than in their wild-type counterparts. CONCLUSIONS: Our data agree with the accumulation of EGFR mutations in a subset of patients with lung cancer. Moreover, we report EGFR gene amplification in EGFR-mutant tumors and a positive correlation between EGFR or KRAS alterations and activation of mTOR signaling. PMID- 16467081 TI - Cytotoxic markers and frequency predict functional capacity of natural killer cells infiltrating renal cell carcinoma. AB - PURPOSE: Renal cell carcinoma harbors high numbers of infiltrating lymphocytes with apparent limited efficacy in tumor control. This study focused on the natural killer (NK) cells infiltrating renal cell carcinoma. EXPERIMENTAL DESIGN: Tumor-infiltrating lymphocytes (TIL) were isolated from renal cell carcinoma and analyzed for NK cell frequency and phenotype (n = 34). NK cells were enriched and tested for effector function. RESULTS: Two renal cell carcinoma subtypes were identified, one containing high (>20% of the lymphocyte population, n = 14), the other low (<20%, n = 20), NK cell numbers. NK cells of both groups were noncytolytic ex vivo but differed in CD16 and cytotoxic effector molecule expression as well as in their capacity to acquire cytotoxic activity: The majority of NK cells from tumors with high NK cell content (high NK-TIL) were CD16(bright), whereas few CD16bright NK cells were found in tumors with low NK cell frequencies (low NK-TIL). The CD16 dichotomy correlated with different capacities to develop cytotoxicity after short-term activation with interleukin-2 ex vivo: Low NK-TIL remained noncytolytic against K562 and unresponsive to signals via the activating receptor NKp46 despite expression of receptor and adaptor molecules. In contrast, high NK-TIL acquired cytotoxic function. As described for peripheral CD16bright NK cells, NK cells from high-NK tumors showed high per cell expression of granzyme A, granzyme B, and perforin. NK cells from low NK-TIL resembled CD16(neg/dim) peripheral NK cells with few cytotoxin+ cells and lower expression of perforin. CONCLUSION: The extent of NK cell infiltration and the expression of markers (CD16 and cytotoxins) predict the functional capacity of NK cells infiltrating renal cell carcinoma and can be used to characterize subgroups of renal cell carcinoma. PMID- 16467082 TI - ZAP-70 expression in normal pro/pre B cells, mature B cells, and in B-cell acute lymphoblastic leukemia. AB - PURPOSE: The ZAP-70 gene is normally expressed in T and natural killer cells, where it is required for the T-cell receptor (TCR) signaling. More recently, it has been described that ZAP-70 contributes to the B-cell development at early stages of B-cell differentiation in mice. The purpose was to investigate the presence of ZAP-70 in normal pro/pre B cells and mature B cells and in tumoral cells from B-acute lymphoblastic leukemias (B-ALL). EXPERIMENTAL DESIGN: ZAP-70 expression was ascertained by flow cytometry, immunofluorescence, Western blot, and quantitative reverse transcription-PCR. Analysis of ZAP-70 and other signaling proteins of the pre-TCR/TCR was done by Western blot. RESULTS: ZAP-70 was expressed in pro/pre B cells but not in normal mature B cells derived from bone marrow, peripheral blood, or tonsil. Among tumoral cells, ZAP-70 was expressed in 56% of B-ALLs with pro/pre B-cell phenotype and in 4 of 6 Burkitt/ALL lymphomas. In B-ALL cells, expression of CD38 protein correlated with ZAP-70 expression (P = 0.05). Mutational analysis of the ZAP-70 gene revealed the absence of mutations in cases lacking ZAP-70 expression. Moreover, other elements of the pre-TCR/TCR signaling pathway, like LAT and Lck, were also found in B-ALL cells. CONCLUSIONS: Among normal B-cell subsets, ZAP-70 was found expressed in normal pro/pre B cells but not in a significant proportion of normal B cells with mature phenotype. Moreover, the presence of ZAP-70 in B-ALLs probably reflects their cellular origin. The lack of ZAP-70 expression in normal mature B cells suggests that its expression in mature-derived neoplasms with different cellular origin, such as Burkitt's lymphoma and chronic lymphocytic leukemia, might be due to an aberrant phenomenon. PMID- 16467083 TI - Homeodomain-interacting protein kinase-2 restrains cytosolic phospholipase A2 dependent prostaglandin E2 generation in human colorectal cancer cells. AB - PURPOSE: Homeodomain-interacting protein kinase-2 (HIPK2), a corepressor for homeodomain transcription factors, is a multifunctional kinase whose role in tumor cell survival is not completely clarified. We addressed whether HIPK2 restrains colon tumorigenesis by turning off cytosolic phospholipase A2 (cPLA2) dependent prostaglandin E2 (PGE2) generation in the light of overwhelming evidence suggesting the contribution of this prostanoid in a variety of cancers. EXPERIMENTAL DESIGN: In the human colorectal cancer cell line, RKO, we studied the effect of RNA interference for HIPK2 (HIPK2i) on prostanoid biosynthesis, both in the absence and in the presence of the cPLA2 inhibitor arachidonyl trifluoromethyl ketone. We evaluated the role of HIPK2 in the cPLA2 gene regulation by reverse transcriptase-PCR, transcriptional activity, and chromatin immunoprecipitation analyses. The involvement of HIPK2 in tumorigenicity in vivo was studied by tumor growth of HIPK2i cells in nude mice. We compared the gene expression of HIPK2 and cPLA2 in human colorectal cancer specimens by reverse transcriptase-PCR. RESULTS: HIPK2 silencing was associated with rousing PGE2 biosynthesis that was profoundly suppressed by the cPLA2 inhibitor. HIPK2 overexpression, along with histone deacetylase-1, inhibited the cPLA2-luc promoter that is strongly acetylated in HIPK2i cells. The tumors derived from HIPK2i cells injected in nude mice showed noticeably increased growth compared with parental cells. HIPK2 mRNA levels were significantly higher in colorectal cancers of patients with familial adenomatous polyposis, which showed undetectable cPLA2 levels compared with sporadic colorectal cancer expressing cPLA2. CONCLUSIONS: Our findings reveal the novel mechanism of HIPK2 to restrain progression of human colon tumorigenesis, at least in part, by turning off cPLA2 dependent PGE2 generation. PMID- 16467084 TI - Purification and characterization of human kallikrein 11, a candidate prostate and ovarian cancer biomarker, from seminal plasma. AB - PURPOSE: Preliminary data suggest that hK11 is a novel serum biomarker for prostate and ovarian cancer. To examine the enzymatic characteristics of hK11, we purified and functionally characterized native hK11 from seminal plasma. EXPERIMENTAL DESIGN: hK11 was purified from seminal plasma by immunoaffinity chromatography and characterized by kinetic analysis, electrophoresis, Western blots, and mass spectrometry. RESULTS: hK11 is present in seminal plasma at concentrations ranging from 2 to 37 microg/mL. Using immunoaffinity chromatography and reverse-phase high-performance liquid chromatography, we purified hK11 to homogeneity. In seminal plasma, hK11 is present as a free enzyme of approximately 40 kDa. About 40% of hK11 is enzymatically active, whereas the rest is inactivated by internal cleavage after Arg156 (Genbank accession no. AF164623), which generates two peptides of approximately 20 kDa, connected by internal disulfide bonds. Purified hK11 possesses trypsin-like activity and cleaves synthetic peptides after arginine but not lysine residues. It does not cleave chymotrypsin substrates. Antithrombin, alpha1-antichymotrypsin, alpha2 antiplasmin, and alpha1-antitrypsin have no effect on hK11 activity and do not form complexes with hK11 in vitro. The strongest inhibitor, APMSF, completely inhibited hK11 activity at a concentration of 2.5 mmol/L. Aprotinin and an hK11 specific monoclonal antibody inhibited hK11 activity up to 40%. Plasmin is a strong candidate for cleaving hK11 at Arg156. CONCLUSION: This is the first report on purification and characterization of native hK11. We speculate that hK11, along with other kallikreins, proteases, and inhibitors, participates in a cascade enzymatic pathway responsible for semen liquefaction after ejaculation. PMID- 16467085 TI - A rapid and sensitive enzymatic method for epidermal growth factor receptor mutation screening. AB - PURPOSE: Mutations in the epidermal growth factor receptor (EGFR) are associated with clinical and radiographic responses to EGFR tyrosine kinase inhibitors gefitinib and erlotinib. Currently available methods of EGFR mutation detection rely on direct DNA sequencing, which requires isolation of DNA from a relatively pure population of tumor cells, cannot be done on small diagnostic specimens, and lack sensitivity. Here we describe the use of a sensitive screening method that overcomes many of these limitations. EXPERIMENTAL DESIGN: We screened 178 non small cell lung cancer specimens for mutations in exons 18 to 21 of EGFR using a DNA endonuclease, SURVEYOR, which cleaves mismatched heteroduplexed DNA. Samples were analyzed by high-performance liquid chromatography on the Transgenomic WAVE HS system. Selected specimens that produced digestion products using SURVEYOR were subsequently reanalyzed by size separation or under partially denaturing conditions, followed by fractionation and sequencing. The specimens included DNA isolated from frozen tumor specimens, dissected formalin-fixed, paraffin-embedded tumor specimens undergoing clinical sequencing, and undissected formalin-fixed, paraffin-embedded specimens. One hundred sixty specimens were independently analyzed using direct DNA sequencing in a blinded fashion. RESULTS: EGFR mutations were detected in 16 of 61 fresh frozen tumor specimens, 24 of 91 dissected formalin-fixed, paraffin-embedded tumor specimens, and 11 of 26 undissected formalin-fixed, paraffin-embedded tumor specimens. Compared with sequencing, the sensitivity and specificity of the present method were 100% and 87%. The positive and negative predictive values were 74% and 100%, respectively. SURVEYOR analysis detected 7 (4%) mutations that were not previously detected by direct sequencing. CONCLUSIONS: SURVEYOR analysis provides a rapid method for EGFR mutation screening with 100% sensitivity and negative predictive value. This unbiased scanning technique is superior to direct sequencing when used with undissected formalin-fixed, paraffin-embedded specimens. PMID- 16467086 TI - The GNAS1 T393C polymorphism predicts survival in patients with clear cell renal cell carcinoma. AB - PURPOSE: G proteins mediate signaling from cell surface receptors to specific intracellular proteins. In vitro cancer cell line studies revealed a link between the Galphas protein and proapoptotic processes. We have recently shown that TT genotypes of the GNAS1 T393C polymorphism display increased transcription of Galphas and a more favorable clinical course in bladder and colorectal cancer patients compared both with TC or CC genotypes. EXPERIMENTAL DESIGN: In the present study, 150 patients with clear cell renal cell carcinoma surgically treated by nephrectomy with curative intent were retrospectively genotyped to elucidate a potential association between T393C genotypes and clinical outcome. RESULTS: The C-allele frequency in the renal cell carcinoma patient group was 0.51, which is not significantly different from that of a healthy blood donor group. Kaplan-Meier curves for tumor progression, development of metastasis, and tumor-related death showed a significant association of the T393C polymorphism with outcome (5-year cancer-specific survival rates: TT, 91%; TC, 81%; CC, 69%; P = 0.015). Multivariate Cox proportional analysis of a 10-year follow-up confirmed the T393C polymorphism as an independent prognostic factor in clear cell renal cell carcinoma. Homozygous CC patients were at highest risk for progression (hazard ratio, 2.48; P = 0.009) or tumor-related death (hazard ratio, 3.15; P = 0.018) compared with T-allele carriers. CONCLUSION: Our results show that besides tumor stage, lymph node status, and tumor grade, the GNAS1 T393C status is a novel independent host factor for disease progression in patients with clear cell renal cell carcinoma and provides further evidence for the T393C polymorphism as a general prognostic tumor marker. PMID- 16467087 TI - Tumor antigen expression in melanoma varies according to antigen and stage. AB - PURPOSE: Melanoma cells express antigens that can induce T-cell and antibody responses. Obtaining a detailed understanding of antigen expression in primary and metastatic melanoma is essential if these molecules are to be useful targets for immunotherapy of melanoma. EXPERIMENTAL DESIGN: Malignant melanomas (n = 586) from 426 patients were typed for antigen expression. Multiple samples were available from 86 individuals, enabling analysis of antigen expression patterns over time. Paraffin-embedded samples were tested by immunohistochemistry for the presence of the differentiation antigens: gp100, Melan-A, tyrosinase, and the "cancer/testis" antigens MAGE-A1, MAGE-A4, and NY-ESO-1. RESULTS: Samples were primary tumors (n = 251), lymph node metastases (n = 174), s.c. metastases (n = 71), and distant metastases (n = 90). The differentiation antigens were strongly expressed in 93% to 95% of tumors regardless of stage. In contrast, the frequency of cancer/testis antigen expression in primary tumors for MAGE-A1, MAGE-A4, and NY-ESO-1 was lower (20%, 9%, and 45%, respectively). MAGE-A1 and MAGE-A4 were acquired with advancing disease (to 51% and 44% in distant metastases, respectively) but not NY-ESO-1, which remained positive in 45%. MAGE-A1 expression was twice as prevalent in ulcerated primaries as in nonulcerated primaries (30% versus 15%; P = 0.006) and in thicker as opposed to thin melanomas (26% versus 10%; P = 0.1). CONCLUSIONS: This large series describes patterns of antigen expression in melanoma and their evolution over time. This will help inform decisions about selection of patients and target antigens for melanoma immunotherapy clinical trials. PMID- 16467088 TI - The cytogenetic relationship between primary and recurrent meningiomas points to the need for new treatment strategies in cases at high risk of relapse. AB - PURPOSE: Recurrence is the major factor influencing the clinical outcome of meningioma patients although the exact relationship between primary and recurrent tumors still needs to be clarified. The aim of the present study is to analyze the cytogenetic relationship between primary and subsequent recurrent meningiomas developed within the same individual. EXPERIMENTAL DESIGN: Multicolor interphase fluorescence in situ hybridization was done for the identification of numerical abnormalities of 12 chromosomes in single-cell suspensions from 59 tumor samples corresponding to 25 recurrent meningioma patients. In 47 of these tumors, the distribution of different tumor cell clones was also analyzed in paraffin embedded tissue sections. In parallel, 132 nonrecurrent cases were also studied. RESULTS: Most recurrent meningiomas showed complex cytogenetic aberrations associated with two or more tumor cell clones in the first tumor analyzed. Interestingly, in most individuals (74%), exactly the same tumor cell clones identified in the initial lesion were also detected in the subsequent recurrent tumor samples. In the recurrent tumor samples of the remaining cases (26%), we observed tumor cell clones related to those detected in the initial lesion but which had acquired one or more additional chromosome aberrations associated with either the emergence of new clones with more complex karyotypes or the disappearance of the most representative clones from the primary lesions. Multivariate analysis of prognostic factors showed that the Maillo et al. prognostic score, based on age of patient, tumor grade, and monosomy 14, together with tumor size was the best combination of independent variables for predicting tumor recurrence at diagnosis. CONCLUSION: Overall, our results indicate that the development of recurrent meningiomas after complete tumor resection is usually due to regrowth of the primary tumor and rarely to the emergence of an unrelated meningioma, underlining the need for alternative treatment strategies in cases at high risk of relapse, particularly those with a high Maillo et al. prognostic score and larger tumors. PMID- 16467089 TI - Molecular subtyping of breast cancer from traditional tumor marker profiles using parallel clustering methods. AB - PURPOSE: Recent small-sized genomic studies on the identification of breast cancer bioprofiles have led to profoundly dishomogenous results. Thus, we sought to identify distinct tumor profiles with possible clinical relevance based on clusters of immunohistochemical molecular markers measured on a large, single institution, case series. EXPERIMENTAL DESIGN: Tumor biological profiles were explored on 633 archival tissue samples analyzed by immunohistochemistry. Five validated markers were considered, i.e., estrogen receptors (ER), progesterone receptors (PR), Ki-67/MIB1 as a proliferation marker, HER2/NEU, and p53 in their original scale of measurement. The results obtained were analyzed by three different clustering algorithms. Four different indices were then used to select the different profiles (number of clusters). RESULTS: The best classification was obtained creating four clusters. Notably, three clusters were identified according to low, intermediate, and high ER/PR levels. A further subdivision in two biologically distinct subtypes was determined by the presence/absence of HER2/NEU and of p53. As expected, the cluster with high ER/PR levels was characterized by a much better prognosis and response to hormone therapy compared to that with the lowest ER/PR values. Notably, the cluster characterized by high HER2/NEU levels showed intermediate prognosis, but a rather poor response to hormone therapy. CONCLUSIONS: Our results show the possibility of profiling breast cancers by means of traditional markers, and have novel clinical implications on the definition of the prognosis of cancer patients. These findings support the existence of a tumor subtype that responds poorly to hormone therapy, characterized by HER2/NEU overexpression. PMID- 16467090 TI - Proteomic analysis of malignant ovarian cancer effusions as a tool for biologic and prognostic profiling. AB - PURPOSE: Malignant epithelial ovarian cancer effusions are important in disease dissemination and clinical outcome. The identification of biochemical events active in effusions may improve our identification and application of targeted therapeutics. EXPERIMENTAL DESIGN: Archival effusion samples for which outcome information was known were studied. Clinical variables were comparable between these groups. Two cohorts of patients with malignant effusion were assessed: those with effusion at presentation (Tap1) or at first recurrence (Tap2). Expression and activated fraction of selected signaling proteins were quantitated on serial protein microarrays using validated antibodies. Proteomic results and clinical variables were analyzed by univariate analysis followed by Cox proportional hazards model analysis. RESULTS: Malignant effusions (>80% malignant cells) were distinguished from benign effusions by higher expression of AKT, activated extracellular signal-regulated kinase, activated (P < or = 0.001) and total cAMP-responsive element binding protein (P = 0.01), and JNK (P = 0.03). Malignant pleural effusions could not be differentiated from ascites by signaling profiles. Both had signal expression clusters for survival, proliferation and metastasis, and injury pathways. Cox proportional hazards model analysis revealed high p38 and pEGFR/EGFR ratio as jointly associated with poor survival in Tap1 cases (both P < or = 0.002). Phospho-JNK quantity was associated with worse outcome in Tap2 patients (P = 0.004), when taking other factors into consideration. CONCLUSIONS: Proliferation, survival, and apoptosis signaling dysregulation can be identified in ovarian cancer effusion samples. Biochemical characterization of clinical effusions may provide either predictive and/or correlative information on patient outcome from which to further understand the mechanisms of effusion development and target clinical intervention. PMID- 16467091 TI - Lymphangiogenesis and angiogenesis in bladder cancer: prognostic implications and regulation by vascular endothelial growth factors-A, -C, and -D. AB - PURPOSE: Lymph vessel density (LVD) and microvessel density (MVD) correlate with the malignant potential of tumors and patient survival. Vascular endothelial growth factors (VEGF)-A, VEGF-C, and VEGF-D could modulate LVD and MVD. We investigated the clinical and prognostic significance of LVD and MVD on lymphangiogenic and angiogenic function of VEGF-A, VEGF-C, and VEGF-D in human bladder cancer. EXPERIMENTAL DESIGN: We reviewed tissue samples from patients with nonmetastatic bladder cancer who had undergone transurethral resections (n = 126). The densities of D2-40-positive vessels (LVD) and CD34-positive vessels (MVD) were measured by a computer-aided image analysis system. Expression of VEGF A, VEGF-C, and VEGF-D was examined by immunohistochemistry; survival analyses and their independent roles were investigated using multivariate analysis models. RESULTS: LVD was associated with tumor grade but not with pT stage. LVD was associated with metastasis-free survival (log rank P = 0.039), but was not an independent prognostic factor. Although MVD affected survival, the combination of high LVD and high MVD in tumors was an independent predictor of metastasis-free survival. Although VEGF-C expression was positively associated with both LVD and MVD, VEGF-D was associated only with LVD. VEGF-A expression was associated with MVD in univariate analysis, however, it was not an independent factor. CONCLUSIONS: Lymphangiogenesis and angiogenesis influence metastasis-free survival, and are regulated by VEGF-C and/or VEGF-D. Our results suggest that LVD and MVD are useful tools for the selection of postoperative management and treatment strategies in patients with bladder cancer. PMID- 16467092 TI - Plasma levels of trefoil factors are increased in patients with advanced prostate cancer. AB - PURPOSE: Through cDNA array analyses and immunohistochemistry on tissue microarrays, trefoil factor 3 (TFF3) was recently shown to be overexpressed in prostate cancer. The purpose of this study was to test the feasibility of using the levels of trefoil factors as a plasma marker for prostate cancer. EXPERIMENTAL DESIGN: In 79 patients with prostate cancer, 23 patients with benign prostatic hyperplasia, and 44 healthy individuals plasma TFF1, TFF2, and TFF3 were determined with ELISAs and compared with clinical stage and prostate specific antigen (PSA) values. Plasma levels of TFF were compared with the immunohistochemical expression of TFF and chromogranin A in 30 prostate cancer tissue samples. RESULTS: Patients with advanced prostate cancer had significantly higher plasma concentrations of TFF1, TFF2, and TFF3 (P < 0.01) compared with patients with localized disease. Using a cutoff of 200 pmol/L, the sensitivity and specificity of plasma TFF3 in differentiating between patients with localized and advanced disease was 74% (59-85%) and 81% (66-91%). Plasma levels of TFF3 were highest in patients with bone metastases (P = 0.008). Patients with serum PSA >10 microg/L had significantly higher plasma TFF3 values than patients with serum PSA <10 microg/L (P = 0.03) and TFF3 levels were higher in patients with Gleason sums of > or = 7 (P = 0.02). Expression of TFF1 and TFF3 determined by immunohistochemistry was increased in patients with prostate cancer but did not correlate with plasma trefoil factor values. CONCLUSIONS: Plasma levels of trefoil factors are increased in patients with advanced prostate cancer. Prospective studies are needed to confirm the predictive utility of trefoil factors in prostate cancer. PMID- 16467093 TI - Optical coherence tomography: real-time imaging of bronchial airways microstructure and detection of inflammatory/neoplastic morphologic changes. AB - PURPOSE: Current diagnostic imaging modalities for human bronchial airways do not possess sufficient resolution and tissue penetration depth to detect early morphologic changes and to differentiate in real-time neoplastic pathology from nonspecific aberrations. Optical coherence tomography (OCT) possesses the requisite high spatial resolution for reproducible delineation of endobronchial wall profiling. EXPERIMENTAL DESIGN: To establish whether OCT could differentiate between the composite microstructural layers of the human airways and simultaneously determine in situ morphologic changes, using a bench-top OCT system, we obtained cross-sectional images of bronchi from 15 patients undergoing lung resections for cancer. All scanned sections underwent subsequent detailed histologic analysis, allowing direct comparisons to be made. RESULTS: OCT imaging enables characterization of the multilayered microstructural anatomy of the airways, with a maximum penetration depth up to 2 to 3 mm and 10-microm spatial resolution. The epithelium, subepithelial components, and cartilage are individually defined. The acquired OCT images closely match histologically defined patterns in terms of structural profiles. Furthermore, OCT identifies in situ morphologic changes associated with inflammatory infiltrates, squamous metaplasia, and tumor presence. CONCLUSIONS: Our results confirm that OCT is a highly feasible optical tool for real-time near-histologic imaging of endobronchial pathology, with potential for lung cancer surveillance applications in diagnosis and treatment. PMID- 16467094 TI - Gene expression profiles of multiple breast cancer phenotypes and response to neoadjuvant chemotherapy. AB - PURPOSE: Breast cancer is a heterogeneous disease, and markers for disease subtypes and therapy response remain poorly defined. For that reason, we employed a prospective neoadjuvant study in locally advanced breast cancer to identify molecular signatures of gene expression correlating with known prognostic clinical phenotypes, such as inflammatory breast cancer or the presence of hypoxia. In addition, we defined molecular signatures that correlate with response to neoadjuvant chemotherapy. EXPERIMENTAL DESIGN: Tissue was collected under ultrasound guidance from patients with stage IIB/III breast cancer before four cycles of neoadjuvant liposomal doxorubicin paclitaxel chemotherapy combined with local whole breast hyperthermia. Gene expression analysis was done using Affymetrix U133 Plus 2.0 GeneChip arrays. RESULTS: Gene expression patterns were identified that defined the phenotypes of inflammatory breast cancer as well as tumor hypoxia. In addition, molecular signatures were identified that predicted the persistence of malignancy in the axillary lymph nodes after neoadjuvant chemotherapy. This persistent lymph node signature significantly correlated with disease-free survival in two separate large populations of breast cancer patients. CONCLUSIONS: Gene expression signatures have the capacity to identify clinically significant features of breast cancer and can predict which individual patients are likely to be resistant to neoadjuvant therapy, thus providing the opportunity to guide treatment decisions. PMID- 16467095 TI - Diffuse mesothelin expression correlates with prolonged patient survival in ovarian serous carcinoma. AB - PURPOSE: Mesothelin is an emerging marker for cancer diagnosis and target-based therapy, yet relatively little is known about the clinical significance of mesothelin expression in tumors. In this study, we correlate mesothelin immunoreactivity to clinicopathologic features in ovarian serous carcinoma. EXPERIMENTAL DESIGN: Mesothelin expression levels were compared among 81 publicly available serial analysis of gene expression (SAGE) libraries of various carcinoma and normal tissue types. Immunohistochemistry using a well characterized mesothelin monoclonal antibody (5B2) was done to evaluate mesothelin expression in 167 high-grade and 31 low-grade ovarian serous carcinomas. Immunohistochemistry staining scores were correlated with patient survival, tumor site, tumor grade, in vitro drug resistance, and differentiation status of tumor cells. RESULTS: SAGE analysis showed that mesothelin was overexpressed in 50% of ovarian and pancreatic carcinomas but rarely in other cancer types, including liver, colon, kidney, prostate, and breast. Mesothelin immunoreactivity (>5% of tumor cells) was present in 55% of ovarian serous carcinomas with no difference in expression between high-grade and low-grade serous tumors (P = 0.82). Based on Kaplan-Meier analysis, we found that a diffuse mesothelin staining (>50% of tumor cells) in primary high-grade ovarian carcinomas correlated significantly with prolonged survival in patients who had advanced-stage disease and had received optimal debulking surgery followed by chemotherapy (P = 0.023). Mesothelin expression did not correlate significantly with patient age, tumor site, in vitro drug resistance, or tumor differentiation status (P > 0.10). CONCLUSION: Our results provided new evidence that mesothelin expression is associated with prolonged survival in patients with high-grade ovarian serous carcinoma. PMID- 16467096 TI - A phase II trial of pemetrexed in advanced breast cancer: clinical response and association with molecular target expression. AB - PURPOSE: This phase II trial of pemetrexed explored potential correlations between treatment outcome (antitumor activity) and molecular target expression. EXPERIMENTAL DESIGN: Chemonaive patients with advanced breast cancer received up to three cycles of pemetrexed 500 mg/m2 (10-minute i.v. infusion) on day 1 of a 21-day cycle, with folic acid and vitamin B12 supplementation. Tumors were surgically removed after the last cycle of pemetrexed as clinically indicated. Biopsies were taken at baseline, 24 hours after infusion in cycle 1, and after cycle 3. RESULTS: Sixty-one women (median age, 46 years; range, 32-72 years) were treated and were evaluable for response. Objective response rate was 31%. Simple logistic regression suggested a potential relationship between mRNA expression of thymidylate synthase (TS) and pemetrexed response (P = 0.103). Based on threshold analysis, patients with "low" baseline TS (< or = 71) were more likely to respond to pemetrexed than patients with "high" baseline TS (>71). Expression of baseline dihydrofolate reductase and glycinamide ribonucleotide formyl transferase tended to be higher in responders but this association was not significant (P > 0.311). TS expression increased significantly between baseline and biopsy 2 (P = 0.004) and dropped to near baseline levels at biopsy 3. Conversely, dihydrofolate reductase and glycinamide ribonucleotide formyl transferase decreased after pemetrexed chemotherapy. CONCLUSIONS: Our results suggest a potential association between "low" pretreatment TS expression levels and response to pemetrexed chemotherapy. Future trials examining expression levels of other genes important to the folate pathway and/or breast cancer may identify a more robust multigene profile that can better predict response to this novel antifolate. PMID- 16467097 TI - Clinical course of patients with non-small cell lung cancer and epidermal growth factor receptor exon 19 and exon 21 mutations treated with gefitinib or erlotinib. AB - PURPOSE: In patients with non-small cell lung cancer (NSCLC), mutations in the epidermal growth factor receptor (EGFR) tyrosine kinase domain have been associated with sensitivity to erlotinib and gefitinib. We undertook this study to explore the relationship between EGFR mutation type and clinical variables, including treatment with gefitinib and erlotinib. EXPERIMENTAL DESIGN: In patients with NSCLC, EGFR exon 19 deletion mutations and EGFR L858R point mutations were analyzed by nonsequencing PCR-based methods from paraffin blocks of tissue obtained before treatment. The results were correlated with clinical information (sex, pathologic subtype, race/ethnicity, treatment, and overall survival). RESULTS: The two most common EGFR mutations were identified in 24% (70 of 291; 95% confidence interval, 26%-38%) of tumors from patients with NSCLC. EGFR mutation was associated with Asian ethnicity (P = 0.0023) and being a "never smoker" (P = 0.0001). Among patients with EGFR mutations, 39% (27 of 70) had EGFR L858R, whereas 61% (43 of 70) had an EGFR exon 19 deletion. After treatment with erlotinib (n = 12) or gefitinib (n = 22), patients with EGFR mutations had a median overall survival of 20 months. After treatment with erlotinib or gefitinib, patients with EGFR exon 19 deletions had significantly longer median survival than patients with EGFR L858R (34 versus 8 months; log-rank P = 0.01). CONCLUSIONS: EGFR mutations in exons 19 or 21 are correlated with clinical factors predictive of response to gefitinib and erlotinib. Those with EGFR exon 19 deletion mutations had a longer median survival than patients with EGFR L858R point mutation. These observations warrant confirmation in a prospective study and exploration of the biological mechanisms of the differences between the two major EGFR mutations. PMID- 16467098 TI - Herceptin sensitizes ErbB2-overexpressing cells to apoptosis by reducing antiapoptotic Mcl-1 expression. AB - PURPOSE: Monoclonal antibodies, such as herceptin and trastuzumab, against the epidermal growth factor receptor ErbB2 (also known as HER2/neu) are an effective therapy for breast cancer patients with overexpression of ErbB2. Herceptin, in combination with standard chemotherapy, such as taxol or etoposide, gives a synergistically apoptotic response in breast tumors. EXPERIMENTAL DESIGN: The mechanism underlying this synergy between chemotherapy and herceptin treatment is not well understood. Herein, we have determined that addition of herceptin, sensitized breast cancer cell lines MDA-MB-231 and MCF-7 to etoposide- or taxol induced apoptosis. RESULTS: This treatment resulted in reduced expression of ErbB2 and the antiapoptotic Bcl-2 family member Mcl-1 in MDA-MB-231 cells. Using antisense oligonucleotides against Mcl-1, MDA-MB-231 cells were rendered sensitive to etoposide-induced apoptosis similar to herceptin, but combined treatment of antisense against Mcl-1 and herceptin failed to give a significant increase in apoptosis. In 29 human breast tumors immunostained for ErbB2 and Mcl 1, we found that when ErbB2 was overexpressed, there was a corresponding increase in Mcl-1 expression. DISCUSSION: Using murine fibroblasts that express human ErbB2, but no other ErbB family member (NE2), these cells showed resistance to both taxol- and etoposide-induced apoptosis compared with parental cells. In addition, NE2 cells preferentially express the antiapoptotic Bcl-2 family member Mcl-1 compared with parental cells, and treatment with herceptin reduces Mcl-1 expression. Taken together, these results suggest that herceptin sensitizes ErbB2 overexpressing cells to apoptosis by reducing antiapoptotic Mcl-1 protein levels. PMID- 16467099 TI - mdr-1 single nucleotide polymorphisms in ovarian cancer tissue: G2677T/A correlates with response to paclitaxel chemotherapy. AB - PURPOSE: P-glycoprotein, encoded by the mdr-1 gene, confers multidrug resistance to a variety of antineoplastic agents, e.g., paclitaxel. Recently, different polymorphisms in the mdr-1 gene have been identified and their consequences for the function of P-glycoprotein, as well as for the treatment response to P glycoprotein substrates, are being clarified. We analyzed the allelic frequencies at polymorphic sites G2677T/A and C3435T in ovarian cancer patients with good or poor response to treatment with paclitaxel in combination with carboplatin in order to evaluate their predictive values. EXPERIMENTAL DESIGN: Fifty-three patients were included in the study; 28 of them had been relapse-free for at least 1 year and 25 had progressive disease or relapsed within 12 months. A reference material consisting of 200 individuals was also analyzed. The genotypes of each single nucleotide polymorphism (SNP) were determined using Pyrosequencing. RESULTS: The G2677T/A SNP was found to significantly correlate with treatment outcome. The probability of responding to paclitaxel treatment was higher in homozygously mutated patients (T/T or T/A; Fisher's exact test; P < 0.05). The frequency of the T or A alleles was also higher in the group of patients who had a good response (P < 0.05). There was also a dose-dependent influence of the number of mutated alleles on the response to paclitaxel treatment (chi2 test for linear-by-linear association; P = 0.03). However, the C3435T SNP was not found to correlate to treatment outcome. CONCLUSIONS: The mdr 1 polymorphism G2677T/A in exon 21 correlates with the paclitaxel response in ovarian cancer and may be important for the function of P-glycoprotein and resistance to paclitaxel and provide useful information for individualized therapy. PMID- 16467100 TI - Phase 1 trial of gefitinib plus sirolimus in adults with recurrent malignant glioma. AB - PURPOSE: To determine the maximum tolerated dose (MTD) and dose-limiting toxicity (DLT) of gefitinib, a receptor tyrosine kinase inhibitor of the epidermal growth factor receptor, plus sirolimus, an inhibitor of the mammalian target of rapamycin, among patients with recurrent malignant glioma. PATIENTS AND METHODS: Gefitinib and sirolimus were administered on a continuous daily dosing schedule at dose levels that were escalated in successive cohorts of malignant glioma patients at any recurrence who were stratified based on concurrent use of CYP3A inducing anticonvulsants [enzyme-inducing antiepileptic drugs, (EIAED)]. Pharmacokinetic and archival tumor biomarker data were also assessed. RESULTS: Thirty-four patients with progressive disease after prior radiation therapy and chemotherapy were enrolled, including 29 (85%) with glioblastoma multiforme and 5 (15%) with anaplastic glioma. The MTD was 500 mg of gefitinib plus 5 mg of sirolimus for patients not on EIAEDs and 1,000 mg of gefitinib plus 10 mg of sirolimus for patients on EIAEDs. DLTs included mucositis, diarrhea, rash, thrombocytopenia, and hypertriglyceridemia. Gefitinib exposure was not affected by sirolimus administration but was significantly lowered by concurrent EIAED use. Two patients (6%) achieved a partial radiographic response, and 13 patients (38%) achieved stable disease. CONCLUSION: We show that gefitinib plus sirolimus can be safely coadministered on a continuous, daily dosing schedule, and established the recommended dose level of these agents in combination for future phase 2 clinical trials. PMID- 16467101 TI - Mannan-MUC1-pulsed dendritic cell immunotherapy: a phase I trial in patients with adenocarcinoma. AB - PURPOSE: Tumor antigen-loaded dendritic cells show promise for cancer immunotherapy. This phase I study evaluated immunization with autologous dendritic cells pulsed with mannan-MUC1 fusion protein (MFP) to treat patients with advanced malignancy. EXPERIMENTAL DESIGN: Eligible patients had adenocarcinoma expressing MUC1, were of performance status 0 to 1, with no autoimmune disease. Patients underwent leukapheresis to generate dendritic cells by culture ex vivo with granulocyte macrophage colony-stimulating factor and interleukin 4 for 5 days. Dendritic cells were then pulsed overnight with MFP and harvested for reinjection. Patients underwent three cycles of leukapheresis and reinjection at monthly intervals. Patients with clinical benefit were able to continue with dendritic cell-MFP immunotherapy. RESULTS: Ten patients with a range of tumor types were enrolled, with median age of 60 years (range, 33-70 years); eight patients were of performance status 0 and two of performance status 1. Dendritic cell-MFP therapy led to strong T-cell IFNgamma Elispot responses to the vaccine and delayed-type hypersensitivity responses at injection sites in nine patients who completed treatments. Immune responses were sustained at 1 year in monitored patients. Antibody responses were seen in three patients only and were of low titer. Side effects were grade 1 only. Two patients with clearly progressive disease (ovarian and renal carcinoma) at entry were stable after initial therapy and went on to further leukapheresis and dendritic cell-MFP immunotherapy. These two patients have now each completed over 3 years of treatment. CONCLUSIONS: Immunization produced T-cell responses in all patients with evidence of tumor stabilization in 2 of the 10 advanced cancer patients treated. These data support further clinical evaluation of this dendritic cell MFP immunotherapy. PMID- 16467102 TI - Combination of p53 cancer vaccine with chemotherapy in patients with extensive stage small cell lung cancer. AB - PURPOSE: The initial goal of this study was to test the immunologic and clinical effects of a new cancer vaccine consisting of dendritic cells (DC) transduced with the full-length wild-type p53 gene delivered via an adenoviral vector in patients with extensive stage small cell lung cancer. EXPERIMENTAL DESIGN: Twenty nine patients with extensive stage small cell lung cancer were vaccinated repeatedly at 2-week intervals. Most of the patients received three immunizations. p53-specific responses were evaluated, and phenotype and function of T cells, DCs, and immature myeloid cells were analyzed and correlated with antigen-specific immune responses. Objective clinical response to vaccination as well as subsequent chemotherapy was evaluated. RESULTS: p53-specific T cell responses to vaccination were observed in 57.1% of patients. Immunologic responses to vaccination were positively associated with a moderate increase in the titer of antiadenovirus antibodies, and negatively with an accumulation of immature myeloid cells. One patient showed a clinical response to vaccination whereas most of the patients had disease progression. However, we observed a high rate of objective clinical responses to chemotherapy (61.9%) that immediately followed vaccination. Clinical response to subsequent chemotherapy was closely associated with induction of immunologic response to vaccination. CONCLUSIONS: This study provides clinical support for an emerging paradigm in cancer immunotherapy, wherein optimal use of vaccination might be more effective, not as a separate modality, but in direct combination with chemotherapy. PMID- 16467103 TI - A metabolic study of patients with lung cancer and hyponatremia of malignancy. AB - PURPOSE: One-third of patients with lung cancer and hyponatremia have no evidence of ectopic arginine vasopressin (AVP) production and the cause of their hyponatremia is not conclusively established. We sought to distinguish patients with hyponatremia caused by elevated AVP versus those with ectopic atrial natriuretic peptide (ANP) via this detailed metabolic study. EXPERIMENTAL DESIGN: We enrolled 24 patients recently diagnosed with lung cancer in a metabolic study in which patients were placed on sodium and fluid restriction for 4 days. Serum electrolytes, osmolality, urine electrolytes and osmolality, plasma AVP, ANP, aldosterone, urinary cyclic AMP and cyclic guanosine 3',5'-monophosphate were measured daily and tumor tissue was obtained to measure ectopic hormone production. We attempted to characterize the pathophysiology of hyponatremia caused by ectopic ANP production in patients with small cell lung cancer (SCLC) and to determine its effect on the aldosterone axis. RESULTS: Seven of the nine patients with SCLC presented with hyponatremia and three had elevated ANP levels at presentation without elevation of AVP. All three patients who presented with hyponatremia and elevated ANP showed a decline in serum sodium following fluid restriction, whereas two patients with SCLC and elevated AVP had normalized serum sodium levels. The combination of hyponatremia and elevated ANP was associated with a persistent natriuresis and inappropriately low aldosterone levels despite sodium restriction, suggesting ANP suppression of the aldosterone axis. CONCLUSIONS: Management of patients with hyponatremia and SCLC should be guided by the knowledge that some patients with SCLC have ectopic production of ANP as the cause of their hyponatremia. PMID- 16467104 TI - 213Bi-[DOTA0, Tyr3]octreotide peptide receptor radionuclide therapy of pancreatic tumors in a preclinical animal model. AB - PURPOSE: The somatostatin analogue [DOTA0, Tyr3]octreotide (DOTATOC) has previously been labeled with low linear energy transfer (LET) beta-emitters, such as 177Lu or 90Y, for tumor therapy. In this study, DOTATOC labeled with the high LET alpha-emitter, 213Bi, was evaluated. EXPERIMENTAL DESIGN: The radiolabeling, stability, biodistribution, toxicity, safety, and therapeutic efficacy of 213Bi DOTATOC (specific activity 7.4 MBq/microg) were investigated. Biodistribution studies to determine somatostatin receptor specificity were done in Lewis rats at 1 and 3 hours postinjection. Histopathology of various organs was used to evaluated toxicity and safety. Therapeutic efficacy of 4 to 22 MBq 213Bi-DOTATOC was determined in a rat pancreatic carcinoma model. RESULTS: Radiolabeling of the 213Bi-DOTATOC was achieved with radiochemical purity >95% and an incorporation yield > or = 99.9%. Biodistribution data showed specific binding to somatostatin receptor-expressing tissues. Administration of free 213Bi, compared with 213Bi DOTATOC, resulted in higher radioactivity accumulation at 3 hours postinjection in the kidneys [34.47 +/- 1.40% injected dose/g (ID/g) tissue versus 11.15 +/- 0.46%, P < 0.0001] and bone marrow (0.31 +/- 0.01% ID/g versus 0.06 +/- 0.02%, P < 0.0324). A significant decrease in tumor growth rate was observed in rats treated with >11 MBq of 213Bi-DOTATOC 10 days postinjection compared with controls (P < 0.025). Treatment with >20 MBq of 213Bi-DOTATOC showed significantly greater tumor reduction when compared with animals receiving <11 MBq (P < 0.02). CONCLUSIONS: 213Bi-DOTATOC showed dose-related antitumor effects with minimal treatment-related organ toxicity. No acute or chronic hematologic toxicities were observed. Mild, acute nephrotoxicity was observed without evidence of chronic toxicity. 213Bi-DOTATOC is a promising therapeutic radiopharmaceutical for further evaluation. PMID- 16467105 TI - Strategies for delaying or treating in vivo acquired resistance to trastuzumab in human breast cancer xenografts. AB - PURPOSE: Acquired resistance to trastuzumab (Herceptin) is common in patients whose breast cancers show an initial response to the drug. The basis of this acquired resistance is unknown, hampering strategies to delay or treat such acquired resistance, due in part to the relative lack of appropriate in vivo tumorigenic models. EXPERIMENTAL DESIGN: We derived an erbB-2-positive variant called 231-H2N, obtained by gene transfection from the highly tumorigenic erbB 2/HER2-negative human breast cancer cell line, MDA-MB-231. Unlike MDA-MB-231, the 231-H2N variants was sensitive to trastuzumab both in vitro and especially in vivo, thus allowing selection of variant resistant to drug treatment in the latter situation after showing an initial response. RESULTS: The growth of established orthotopic tumors in severe combined immunodeficient mice was blocked for 1 month by trastuzumab, after which rapid growth resumed. These relapsing tumors were found to maintain resistance to trastuzumab, both in vitro and in vivo. We evaluated various therapeutic strategies for two purposes: (a) to delay such tumor relapses or (b) to treat acquired trastuzumab resistance once it has occurred. With respect to the former, a daily oral low-dose metronomic cyclophosphamide regimen was found to be particularly effective. With respect to the latter, an anti-epidermal growth factor receptor antibody (cetuximab) was effective as was the anti-vascular endothelial growth factor (anti-VEGF) antibody bevacizumab, which was likely related to elevated levels of VEGF detected in trastuzumab-resistant tumors. CONCLUSIONS: Our results provide a possible additional rationale for combined biological therapy using drugs that target both erbB-2/HER2 and VEGF and also suggest the potential value of combining less toxic metronomic chemotherapy regimens not only with targeted antiangiogenic agents but also with other types of drug such as trastuzumab. PMID- 16467106 TI - Tumor vascular permeabilization by vascular-targeting photosensitization: effects, mechanism, and therapeutic implications. AB - PURPOSE: Loss of vascular barrier function has been observed shortly following vascular-targeting photodynamic therapy. However, the mechanism involved in this event is still not clear, and the therapeutic implications associated with this pathophysiologic change have not been fully explored. EXPERIMENTAL DESIGN: The effect of vascular-targeting photodynamic therapy on vascular barrier function was examined in both s.c. and orthotopic MatLyLu rat prostate tumor models and endothelial cells in vitro, using photosensitizer verteporfin. Vascular permeability to macromolecules (Evans blue-albumin and high molecular weight dextran) was assessed with dye extraction (ex vivo) and intravital microscopy (in vivo) methods. Intravital microscopy was also used to monitor tumor vascular functional changes after vascular-targeting photodynamic therapy. The effects of photosensitization on monolayer endothelial cell morphology and cytoskeleton structures were studied with immunofluorescence staining. RESULTS: Vascular targeting photodynamic therapy induced vascular barrier dysfunction in the MatLyLu tumors. Thus, tumor uptake of macromolecules was significantly increased following photodynamic therapy treatments. In addition to vascular permeability increase, blood cell adherence to vessel wall was observed shortly after treatment, further suggesting the loss of endothelial integrity. Blood cell adhesion led to the formation of thrombi that can occlude blood vessels, causing vascular shutdown. However, viable tumor cells were often detected at tumor periphery after vascular-targeting photodynamic therapy. Endothelial cell barrier dysfunction following photodynamic therapy treatment was also observed in vitro by culturing monolayer endothelial cells on Transwell inserts. Immunofluorescence study revealed microtubule depolymerization shortly after photosensitization treatment and stress actin fiber formation thereafter. Consequently, endothelial cells were found to retract, and this endothelial morphologic change led to the formation of intercellular gaps. CONCLUSIONS: Vascular-targeting photodynamic therapy permeabilizes blood vessels through the formation of endothelial intercellular gaps, which are likely induced via endothelial cell microtubule depolymerization following vascular photosensitization. Loss of endothelial barrier function can ultimately lead to tumor vascular shutdown and has significant implications in drug transport and tumor cell metastasis. PMID- 16467107 TI - The schedule-dependent effects of the novel antifolate pralatrexate and gemcitabine are superior to methotrexate and cytarabine in models of human non Hodgkin's lymphoma. AB - PURPOSE: Methotrexate is known to synergize with cytarabine [1-beta-D arabinofuranosylcytosine (ara-C)] in a schedule-dependent manner. The purpose of this article is to compare and contrast the activity of pralatrexate (10 propargyl-10-deazaminopterin)/gemcitabine to the standard combination of methotrexate/ara-C and to determine if schedule dependency of this combination is important in lymphoma. EXPERIMENT DESIGN: Cytotoxicity assays using the standard trypan blue exclusion assay were used to explore the in vitro activity of pralatrexate and gemcitabine against a panel of lymphoma cell lines. Both severe combined imunodeficient beige and irradiated nonobese diabetic/severe combined imunodeficient mouse xenograft models were used to compare and contrast the in vivo activity of these combinations as a function of schedule. In addition, apoptosis assays were conducted. RESULTS: Compared with methotrexate-containing combinations, pralatrexate plus gemcitabine combinations displayed improved therapeutic activity with some schedule dependency. The combination of pralatrexate and gemcitabine was superior to any methotrexate and ara-C combination in inducing apoptosis and in activating caspase-3. In vivo, the best therapeutic effects were obtained with the sequence of pralatrexate --> gemcitabine. Complete remissions were only appreciated in animals receiving pralatrexate followed by gemcitabine. CONCLUSIONS: These data show that the combination of pralatrexate followed by gemcitabine was superior to methotrexate/ara-C in vitro and in vivo, and was far more potent in inducing apoptosis in a large B-cell lymphoma. These data provide strong rationale for further study of this combination in lymphomas where methotrexate and ara-C are used. PMID- 16467108 TI - Lovastatin protects human endothelial cells from killing by ionizing radiation without impairing induction and repair of DNA double-strand breaks. AB - PURPOSE: 3-hydroxy-3-methylglutaryl CoA reductase inhibitors (statins) are frequently used lipid-lowering drugs. Moreover, they are reported to exert pleiotropic effects on cellular stress responses, proliferation, and apoptosis. Whether statins affect the sensitivity of primary human cells to ionizing radiation (IR) is still unknown. The present study aims at answering this question. EXPERIMENTAL DESIGN: The effect of lovastatin on IR-provoked cytotoxicity was analyzed in primary human umbilical vein endothelial cells (HUVEC). To this end, cell viability, proliferation, and apoptosis as well as DNA damage-related stress responses were investigated. RESULTS: The data show that lovastatin protects HUVEC from IR-induced cell death. Lovastatin did not confer radioresistance to human fibroblasts. The radioprotective, antiapoptotic effect of lovastatin was observed at low, physiologically relevant dose level (1 micromol/L). Lovastatin affected various IR-induced stress responses in HUVEC: It attenuated the increase in p53/p21 protein level and impaired the activation of nuclear factor-kappaB, Chk-1, and Akt kinase but did not inhibit extracellular signal-regulated kinase activation. Exposure of HUVEC to IR did not change the level of Bax and Bcl-2 and did not cause activation of caspase-3, indicating that radioprotection by lovastatin does not depend on the modulation of the mitochondrial death pathway. Also, IR-induced DNA double-strand break formation and repair were not influenced by lovastatin. CONCLUSIONS: The data show that lovastatin has multiple inhibitory effects on IR-stimulated DNA damage-dependent stress responses in HUVEC. Because lovastatin causes radioresistance, it might be useful in the clinic for attenuating side effects of radiation therapy that are related to endothelial cell damage. PMID- 16467109 TI - Histone deacetylase inhibitor-mediated radiosensitization of human cancer cells: class differences and the potential influence of p53. AB - Histone deacetylase inhibitors (HDI) are emerging as potentially useful components of the anticancer armamentarium and as useful tools to dissect mechanistic pathways. HDIs that globally inhibit histone deacetylases (HDAC) have radiosensitizing effects, but the relative contribution of specific HDAC classes remains unclear. Newly characterized HDIs are now available that preferentially inhibit specific HDAC classes, including SK7041 (inhibits class I HDACs) and splitomicin (inhibits class III HDACs). We investigated in human cancer cells the relative radiosensitizations that result from blocking specific HDAC classes. We found that trichostatin A (TSA; inhibitor of both class I and II HDACs) was the most effective radiosensitizer, followed by the class I inhibitor SK7041, whereas splitomicin (inhibitor of class III) had least effect. Interestingly, radiosensitization by TSA in cell lines expressing p53 was more pronounced than in isogenic lines lacking p53. Radiosensitization of cells expressing p53 by TSA was reduced by pifithrin-alpha, a small-molecule inhibitor of p53. In contrast, the radiosensitization by TSA of cells expressing low levels of p53 was enhanced by transfection of wild-type p53-expressing vector or pretreatment with leptomycin B, an inhibitor of nuclear export that increased intracellular levels of p53. These effects on radiosensitization were respectively muted or not seen in cells treated with SK7041 or splitomicin. To our knowledge, this may be among the first systematic investigations of the comparative anticancer effects of inhibiting specific classes of HDACs, with results suggesting differences in the degrees of radiosensitization, which in some cell lines may be influenced by p53 expression. PMID- 16467111 TI - Risk stratification of colon carcinogenesis through enhanced backscattering spectroscopy analysis of the uninvolved colonic mucosa. AB - INTRODUCTION: Our group has been interested in applying advances in biomedical optics to colorectal cancer risk stratification. Through a recent technological breakthrough, we have been able to harness information from enhanced backscattering spectroscopy, an optics phenomenon that allows quantitative, depth selective analysis of the epithelial microscale/nanoscale architecture. In the present study, we investigated the ability of enhanced backscattering analysis of the preneoplastic mucosa to predict risk of colon carcinogenesis. METHODS: Enhanced backscattering analysis was done on intestinal mucosa at preneoplastic time points from two experimental models of colorectal cancer: the azoxymethane treated rat and the multiple intestinal neoplasia (MIN) mouse. Data were analyzed using two previously validated spectral markers: spectral slope and principle components. We then did a pilot study on mucosal biopsies from 63 subjects undergoing screening colonoscopy. RESULTS: In the azoxymethane-treated rat, when compared with saline-treated controls, significant changes in the enhanced backscattering markers were observed as early as 2 weeks after azoxymethane treatment (before the development of aberrant crypt foci and adenomas). Enhanced backscattering markers continued to progress over time in a manner consonant with future neoplasia. These data were replicated in the preneoplastic MIN mouse mucosa. In humans, spectral slopes in the endoscopically normal cecum, midtransverse colon, and rectum were markedly reduced in patients harboring adenomas when compared with those who were neoplasia free. CONCLUSIONS: We show, for the first time, that enhanced backscattering analysis of an aliquot of uninvolved mucosa has the potential for predicting neoplastic risk throughout the colon in both experimental colorectal cancer models and humans. PMID- 16467110 TI - BMS-345541 targets inhibitor of kappaB kinase and induces apoptosis in melanoma: involvement of nuclear factor kappaB and mitochondria pathways. AB - PURPOSE: Constitutive activation of inhibitor of kappaB kinase (IKK) confers melanoma resistance to apoptosis and chemotherapy. Whether IKK is able to serve as a therapeutic target in melanoma is unknown. We explored the possibility of exploiting IKK as a therapeutic target in melanoma by using BMS-345541, a novel compound with a highly selective IKKbeta inhibitory activity, to trigger melanoma cell apoptosis. EXPERIMENTAL DESIGN: Three human melanoma cell lines (SK-MEL-5, Hs 294T, and A375), all of which have high constitutive IKK activities, served as in vitro and in vivo melanoma models for treatment with BMS-345541. Two known antitumor drugs (temozolomide and bortezomib) were used as parallel controls for evaluation of the therapeutic efficiency and toxicity of BMS-345541. The effects of BMS-345541 on nuclear factor kappaB (NF-kappaB) signaling and on the apoptosis machinery were investigated. RESULTS: Inhibition of constitutive IKK activity by BMS-345541 resulted in the reduction of NF-kappaB activity, CXCL1 chemokine secretion by cultured melanoma cells and melanoma cell survival in vitro and in vivo. The effect of BMS-345541 on tumor cell growth was through mitochondria mediated apoptosis, based on the release of apoptosis-inducing factor, dissipation of mitochondrial membrane potential, and reduced ratio of B cell lymphoma gene-2 (Bcl-2)/Bcl-associated X protein (Bax) in mitochondria. The BMS 345541 execution of apoptosis was apoptosis-inducing factor-dependent, but largely caspase-independent. CONCLUSION: BMS-345541 down-regulation of IKK activity results in mitochondria-mediated apoptosis of tumor cells because the programmed cell death machinery in melanoma cells is highly regulated by NF kappaB signaling. Therefore, IKK may serve as a potential target for melanoma therapy. PMID- 16467112 TI - Chemoprevention of skin carcinogenesis by phenylretinamides: retinoid receptor independent tumor suppression. AB - Fenretinide [N-(4-hydroxyphenyl)retinamide or 4-HPR] is a synthetic retinoid analogue with antitumor and chemopreventive activities. N-(4 Methoxyphenyl)retinamide (4-MPR) is the most abundant metabolite of 4-HPR detected in human serum following 4-HPR therapy. We have shown in in vitro studies that 4-HPR and 4-MPR can act independent of the classic nuclear retinoid receptor pathway and that 4-HPR, but not 4-MPR, can also activate nuclear retinoid receptors. In this study, we have compared the chemopreventive effects of topically applied 4-HPR and 4-MPR with the primary biologically active retinoid, all-trans retinoic acid (ATRA), in vivo in the mouse skin two-stage chemical carcinogenesis model. All three retinoids suppressed tumor formation but the effect of 4-HPR and 4-MPR, and not of ATRA, was sustained after their discontinuation. The tumor-suppressive effects of 4-HPR and 4-MPR were quantitatively and qualitatively similar, suggesting that the two may be acting through the same retinoid receptor-independent mechanism(s). We further explored this effect in vitro by analyzing primary cultures of mouse keratinocytes treated with the same retinoids. All three could induce apoptosis with a 48-hour treatment and only ATRA and 4-HPR induced an accumulation of cells in the G1 phase of the cell cycle. This finding is consistent with our previous results showing that the effects of phenylretinamides on the cell cycle are retinoid receptor dependent whereas apoptosis induction is not. A microarray-based comparison of gene expression profiles for mouse skin treated with 12-O tetradecanoylphorbol-13-acetate (TPA) alone and TPA + 4-HPR or TPA + 4-MPR reveals a high degree of coincidence between the genes regulated by the two phenylretinamides. We propose that 4-HPR may exert therapeutic and chemopreventive effects by acting primarily through a retinoid receptor independent mechanism(s) and that 4-MPR may contribute to the therapeutic effect of 4-HPR by acting through the same retinoid receptor-independent mechanism(s). PMID- 16467113 TI - 2-Methoxyestradiol inhibits prostate tumor development in transgenic adenocarcinoma of mouse prostate: role of tumor necrosis factor-alpha-stimulated gene 6. AB - PURPOSE: 2-Methoxyestradiol, an estrogenic metabolite, is in clinical trials for the treatment of hormone-refractory prostate cancer. However, neither the chemopreventive role nor the mechanism of 2-methoxyestradiol-induced biological activities is fully understood. EXPERIMENTAL DESIGN: Eight- and 24-week-old transgenic adenocarcinoma of mouse prostate (TRAMP) mice were fed a diet containing 50 mg 2-methoxyestradiol/kg body weight for 16 and 8 weeks, respectively. Chemopreventive efficacy was evaluated by magnetic resonance imaging, determining the prostate-seminal vesicle complex volume and histologic analysis of prostate tumor or tissue. Tumor invasion assays were used to show the role of tumor necrosis factor-alpha-stimulated gene (TSG-6), a 2-methoxyestradiol up-regulated gene identified by DNA array analysis. Expression of TSG-6 was analyzed in a human tissue array containing different grades of prostate tumors. RESULTS: Dietary administration of 2-methoxyestradiol prevented the development of preneoplastic lesions independent of progression stage. TSG-6 was low or undetectable in prostate cancer cells (LNCaP, PC-3, and DU145) and TRAMP tumors but up-regulated in response to 2-methoxyestradiol. Immunohistochemistry of the human prostate tumor array showed a decrease in TSG-6-positive cells with increasing grade relative to normal prostate (P = 0.0001). Although overexpression of TSG-6 inhibited invasion of androgen-independent cells (P = 0.007), antisense TSG-6 reversed this effect. CONCLUSIONS: To the best of our knowledge, this is the first report showing the potential of 2-methoxyestradiol as a chemopreventive agent. We have also identified TSG-6 as a potential marker that could be used for early diagnosis and prognosis of cancerous or precancerous lesions. PMID- 16467114 TI - High levels of aberrant DNA methylation in Helicobacter pylori-infected gastric mucosae and its possible association with gastric cancer risk. AB - INTRODUCTION: Risk prediction of gastric cancers is important to implement appropriate screening procedures. Although aberrant DNA methylation is deeply involved in gastric carcinogenesis, its induction by Helicobacter pylori, a strong gastric carcinogen, is unclear. Here, we analyzed the effect of H. pylori infection on the quantity of methylated DNA molecules in noncancerous gastric mucosae and examined its association with gastric cancer risk. EXPERIMENTAL DESIGN: Gastric mucosae were collected from 154 healthy volunteers (56 H. pylori negative and 98 H. pylori positive) and 72 cases with differentiated-type gastric cancers (29 H. pylori negative and 43 H. pylori positive) by endoscopy. The numbers of DNA molecules methylated and unmethylated for eight regions of seven CpG islands (CGI) were quantified by quantitative PCR after bisulfite modification, and fractions of methylated molecules (methylation levels) were calculated. RESULTS: Among healthy volunteers, methylation levels of all the eight regions were 5.4- to 303-fold higher in H. pylori positives than in H. pylori negatives (P < 0.0001). Methylation levels of the LOX, HAND1, and THBD promoter CGIs and p41ARC exonic CGI were as high as 7.4% or more in H. pylori positive individuals. Among H. pylori-negative individuals, methylation levels of all the eight regions were 2.2- to 32-fold higher in gastric cancer cases than in age-matched healthy volunteers (P < or = 0.01). Among H. pylori-positive individuals, methylation levels were highly variable, and that of only HAND1 was significantly increased in gastric cancer cases (1.4-fold, P = 0.02). CONCLUSIONS: It was indicated that H. pylori infection potently induces methylation of CGIs to various degrees. Methylation levels of specific CGIs seemed to reflect gastric cancer risk in H. pylori-negative individuals. PMID- 16467115 TI - Proceedings of the Fifth International Conference on Recent Advances and Future Directions in Endocrine Therapy for Breast Cancer: conference summary statement. PMID- 16467116 TI - Novel mechanisms of resistance to endocrine therapy: genomic and nongenomic considerations. AB - Selective estrogen receptor (ER) modulators have been the most commonly used neoadjuvant therapy for hormone-dependent breast cancer. However, resistance to endocrine therapy, either inherent or acquired during treatment, presents a major challenge in disease management. The causes of resistance to hormone therapy are not well understood and are the subject of active investigation. It is increasingly clear that decreasing sensitivity of ER-positive breast cancer cells to antiestrogens is caused by several factors. Cross talk between ER and growth factor signaling has emerged as a critical factor in endocrine resistance. Here, we present evidence that receptor tyrosine kinase signaling also plays a role in resistance by controlling the subcellular localization of ER signaling components. Localization of ER in either the nuclear or cytoplasmic compartments has functional implications. Recent work suggests that dynein light chain 1, a recently identified substrate of p21-activated kinase 1, modulates ER transactivation functions through a novel ER coactivator function. Likewise, receptor tyrosine kinase signaling can also alter the expression of ER coregulators such as metastasis-associated antigen 1, leading to hormonal independence. Furthermore, proline-, glutamic acid-, leucine-rich protein 1, an ER coactivator involved in both genomic and nongenomic signaling pathways, is activated by epidermal growth factor receptor and plays a prominent role in resistance to tamoxifen. These recent advances suggest new targeted therapeutic approaches that may lead to either reversion or prevention of endocrine resistance in breast tumors. PMID- 16467118 TI - Progesterone receptor loss correlates with human epidermal growth factor receptor 2 overexpression in estrogen receptor-positive breast cancer. AB - Response to endocrine therapy in breast cancer correlates with estrogen receptor (ER) and progesterone receptor (PR) status. It was originally hypothesized that the ability of PR to predict response to endocrine therapy was due to the fact that PR is an estrogen-regulated gene and that its levels represented a marker of functional ER activity. However, it is now known that loss of PR can occur via multiple mechanisms, many of which do not include ER function, e.g., hypermethylation of the PR promoter and loss of heterozygosity of the PR gene. We have shown that growth factor signaling pathways can directly down-regulate PR levels via the phosphatidylinositol 3'-kinase (PI3K)/Akt/mTOR pathway, and that this can occur independent of ER. For example, overexpression of myr-Akt in MCF-7 cells causes complete loss of PR protein and mRNA but does not reduce ER levels or activity, thus generating ER+/PR- MCF-7 cells. Therefore, the absence of PR may not simply reflect a lack of ER activity but rather may reflect hyperactive cross-talk between ER and growth factor signaling pathways. Consistent with this hypothesis, several recent clinical studies have found that ER+/PR- breast cancers overexpress human epidermal growth factor receptor (HER) 1 and HER2 compared with ER+/PR+ breast cancers. Although HER receptors can lower ER levels, one study showed that loss of PR correlated with high HER2 levels in a multivariate analysis. Furthermore, loss of PTEN, a negative regulator of the PI3K/Akt signaling pathway, has been shown to be associated with specific loss of PR and no change in ER levels. Given the well-recognized resistance of ER+/PR- breast cancer to antiestrogens, more studies are needed to better understand the etiology of ER+/PR- breast cancer, particularly the analysis of other growth factor receptors and their downstream signaling intermediates with respect to PR status. PMID- 16467117 TI - ErbB receptor signaling and therapeutic resistance to aromatase inhibitors. AB - We have investigated the effect of HER-2 overexpression on resistance to the aromatase inhibitor letrozole in MCF-7 breast cancer cells stably expressing cellular aromatase (MCF-7/CA). MCF-7/CA cells overexpressing HER-2 showed a >2 fold increase in estrogen receptor (ER)-mediated transcriptional reporter activity upon treatment with androstenedione compared with vector-only control MCF-7/CA cells. Co-treatment with letrozole did not abrogate androstenedione induced transcription and cell proliferation in HER-2-overexpressing cells. Chromatin immunoprecipitation assays using cross-linked protein-DNA from MCF 7/CA/HER-2 cells indicated ligand-independent association of the ERalpha coactivators AIB-1 and CBP to the promoter region of the estrogen-responsive pS2 gene. Upon treatment with androstenedione, there were increased associations of AIB1 and CBP with the pS2 promoter in the HER-2-overexpressing compared with control MCF-7/CA cells. These results suggest that ligand-independent recruitment of coactivator complexes to estrogen-responsive promoters as a result of HER-2 overexpression may play a role in the development of letrozole resistance. PMID- 16467119 TI - Methods for gene expression profiling in clinical trials of adjuvant breast cancer therapy. AB - Although endocrine therapy is highly effective in the treatment of endocrine receptor-positive breast cancer, chemotherapy has been shown to provide clinical benefit when added to tamoxifen. However, baseline risk after tamoxifen treatment is so low, especially in patients who are axillary node negative, that significant overtreatment will result if chemotherapy is given to every patient. Robust prognostic and predictive markers need to be developed to identify those at high risk of treatment failure. Although comprehensive gene expression profiling methods do offer promise, they require fresh or frozen tumor samples. To take advantage of existing archived tissue blocks with clinical follow-up collected from finished clinical trials, such as National Surgical Adjuvant Breast and Bowel Project trials B-20 and B-14, technologies that allow interrogation of archived blocks for gene expression profiling need to be realized. Recent developments in gene expression profiling technologies are discussed with their implications in clinical management of endocrine receptor positive breast cancer. PMID- 16467121 TI - Adjuvant endocrine therapy for postmenopausal women with early breast cancer. AB - Results from multiple clinical trials involving aromatase inhibitors have added to the knowledge base relating to endocrine therapy of postmenopausal women with hormone receptor-positive early breast cancer. In the extended adjuvant setting, data from the Austrian Breast and Colorectal Cancer Study Group 6a trial showed an advantage for anastrozole following 5 years of tamoxifen treatment, consistent with the more robust MA.17 trial that examined letrozole versus placebo following 5 years of tamoxifen treatment. The combined analysis of the Austrian Breast and Colorectal Cancer Study Group 8 trial and the German Arimidex Nolvadex 95 trial, plus the Italian Tamoxifen Anastrozole trial, have shown the advantage of switching to anastrozole over continuing the tamoxifen to complete the full 5 years of adjuvant therapy. These trials support the previously reported larger and double-blind Intergroup Exemestane Study. The Arimidex, Tamoxifen, Alone or in Combination trial now has data out to 68 months of median follow-up showing the maintenance of a significant advantage of anastrozole over tamoxifen for disease-free survival. In this initial treatment setting, the Breast International Group 1-98 trial recently showed a significant advantage for letrozole over tamoxifen. The current debate is centered on whether the optimal strategy is to give an aromatase inhibitor initially or after several years of tamoxifen treatment. Multiple important questions remain, including the predictive value of molecular markers such as progesterone receptor, the optimal duration of aromatase inhibitor therapy, the long-term adverse effects, and the relative efficacy and toxicity of the different aromatase inhibitors. PMID- 16467120 TI - Proliferation and apoptosis as markers of benefit in neoadjuvant endocrine therapy of breast cancer. AB - The study of changes in proliferation as a marker of treatment benefit during presurgical endocrine treatment of breast cancer has become increasingly popular, particularly using the nuclear marker Ki67, and holds the potential for prioritizing new treatments for full clinical development. There are weakly significant relationships between Ki67 change and clinical response that differ according to data handling. In the neoadjuvant Immediate Preoperative Anastrozole, Tamoxifen, or Combined with Tamoxifen trial, suppression of Ki67 at both 2 and 12 weeks was greater with the aromatase inhibitor anastrozole than with either tamoxifen or the combination of anastrozole and tamoxifen. We report here that absolute values of Ki67 after 2 weeks were also significantly lower with anastrozole than with tamoxifen and the combination. This indicates that it may be possible to make such comparisons using surgical samples only. We argue that these changes in proliferation and concurrent changes in apoptosis may be expected to be more predictive of adjuvant benefit from endocrine therapy than clinical response. PMID- 16467122 TI - Anastrozole as an adjuvant endocrine treatment for postmenopausal patients with breast cancer: emerging data. AB - The Arimidex, Tamoxifen, Alone or in Combination (ATAC) trial compared the efficacy and safety of anastrozole versus tamoxifen versus the combination as initial adjuvant treatment for early breast cancer in over 9,000 postmenopausal women. Analyses at 33 and 47 months median follow-up showed that anastrozole significantly prolonged disease-free survival and time to recurrence and reduced the incidence of contralateral breast cancer compared with tamoxifen. Results of the completed treatment analysis at 68 months median follow-up confirmed the earlier findings, showing that the absolute difference in disease-free survival continued to increase beyond completion of treatment. Mature safety data from the ATAC trial show that, overall, anastrozole has a favorable safety profile compared with tamoxifen. In the absence of current data on further follow-up or the outcome of trials investigating proactive sequencing of endocrine therapies, we present a model based on several trials, including ATAC. This model suggests that using an aromatase inhibitor as initial adjuvant therapy is a better option than switching to an aromatase inhibitor after >/=2 years of tamoxifen. The relative toxicities of the three approved third-generation aromatase inhibitors, anastrozole, letrozole, and exemestane, are discussed. These data suggest that long-term safety profiles may differ between aromatase inhibitors, although comprehensive comparative data for letrozole and exemestane versus tamoxifen are lacking. PMID- 16467123 TI - Is there a role for adjuvant tamoxifen in progesterone receptor-positive breast cancer? An in silico clinical trial. AB - PURPOSE: Subset analysis from the Arimidex, Tamoxifen, Alone or in Combination trial, a major adjuvant therapy trial, suggests that progesterone receptor negative (PR-) cases may derive greater benefit from aromatase inhibitor compared with tamoxifen than PR+ cases. We postulated that estrogen receptor-positive (ER+)/PR+ patients might do as well or better by starting on tamoxifen and later switching to an aromatase inhibitor rather than by starting on an aromatase inhibitor as initial therapy. EXPERIMENTAL DESIGN: We constructed a computer model using retrospective data to approximate exponential failure rates for PR+ and PR- in tamoxifen-treated and tamoxifen-untreated patients, adding the assumptions that about half of patients are cured at surgery and that approximately 20% of postmenopausal ER+ early breast cancer cases are PR-. This model provided a very good approximation to the published overview data and to the clinical trials. We then used the failure rates to generate relapse times for a large number of cases (n = 50,000) for each treatment scenario. RESULTS: In PR- cases, initial therapy with an aromatase inhibitor is superior to tamoxifen and this advantage can never be made up by switching. In PR+ cases, tamoxifen is only modestly inferior to aromatase inhibitor at the outset, and after switching to an aromatase inhibitor at 3 or 5 years the tamoxifen relapse-free survival curve catches up and then begins to surpass the aromatase inhibitor curve at 7.5 or 12 years, respectively. DISCUSSION: Although our in silico trial is based on many assumptions, it closely approximates results of the published trials and, therefore, suggests that an in vivo comparison in ER+/PR+ patients of aromatase inhibitor versus tamoxifen followed by aromatase inhibitor may be worth considering. PMID- 16467124 TI - Managing patients on endocrine therapy: focus on quality-of-life issues. AB - PURPOSE: To review the health-related quality of life (QOL) of women treated with adjuvant hormonal therapy. EXPERIMENTAL DESIGN: To review the limited QOL data from randomized trials of tamoxifen versus placebo and ovarian ablation versus none. To discuss QOL results from randomized trials of aromatase inhibitors compared with tamoxifen or placebo for adjuvant therapy of postmenopausal women with estrogen receptor-positive and/or progesterone receptor-positive breast cancer. RESULTS: QOL is generally good in up to 3 years of follow-up with either tamoxifen or aromatase inhibitors. Vasomotor and sexual complaints remain problematic, however, in only a small proportion of women. There are fewer data regarding the QOL effects of ovarian ablation, which may nonetheless be more substantial. CONCLUSION: Tamoxifen and aromatase inhibitors cause specific vasomotor or gynecologic symptoms, which may affect sexual function. However, clinical benefits of these agents are generally achieved without major detrimental effect on overall QOL. PMID- 16467125 TI - Clinical efforts to combine endocrine agents with targeted therapies against epidermal growth factor receptor/human epidermal growth factor receptor 2 and mammalian target of rapamycin in breast cancer. AB - Enhancing the benefit of endocrine therapy by overcoming de novo or acquired resistance remains an important goal in systemic breast cancer therapy. Progress continues to be made in elucidating the molecular pathways by which estrogen receptor-positive breast cancer cells escape from endocrine therapy. The increasing recognition of the roles of epidermal growth factor receptor (EGFR) and human EGFR2 in cross-talk activation of estrogen receptor signaling has led to studies aimed at identifying whether small-molecule tyrosine kinase inhibitors targeted against these receptors give additive or synergistic effects when combined with endocrine agents. Activation of the phosphatidylinositol-3-OH kinase/Akt pathway has also been associated with resistance to either tamoxifen or estrogen deprivation, and preclinical studies have shown that the mammalian target of rapamycin antagonist temsirolimus can restore endocrine sensitivity in breast cancer cells. Randomized phase II trials of aromatase inhibitors combined with EGFR/human EGFR2 tyrosine kinase inhibitors or mammalian target of rapamycin antagonists have been completed in both the neoadjuvant and advanced breast cancer settings. Larger phase III trials with both approaches are now in progress and have been powered to detect whether either strategy can significantly prolong time to disease progression compared with endocrine therapy alone. The correlation of molecular and clinical results from these ongoing studies will be important to establish appropriate biological variables for selecting those patients who may benefit most from this combined approach. PMID- 16467126 TI - Tubular kidney injury molecule-1 in protein-overload nephropathy. AB - Kim-1, a recently discovered membrane protein, is undetectable in normal kidneys but markedly induced in proximal tubules after ischemic and toxic injury. The function of Kim-1 is unclear, but it is implicated in damage/repair processes. The Kim-1 ectodomain is cleaved by metalloproteinases and detectable in urine. We studied Kim-1 in a nontoxic, nonischemic, model of tubulointerstitial damage caused by acute proteinuria. Uninephrectomized (NX) rats received daily (ip) injections of 2 g BSA (NX+BSA, n = 12) or saline (NX, n = 6) for 3 wk. Kidneys were stained for various damage markers by immunohistochemistry (IHC). Kim-1 mRNA (RT-PCR, in situ hybridization), protein (IHC, Western blotting), and urinary Kim 1 (Luminex) were determined. Spatial relations between Kim-1 and other damage markers were studied by double labeling IHC. NX+BSA rats developed massive proteinuria (1,217 +/- 313 vs. 18 +/- 2 mg/day in NX, P < 0.001) and significant renal damage. Kim-1 mRNA was upregulated eightfold in NX+BSA (ratio Kim-1/beta actin, 4.08 +/- 2.56 vs. 0.52 +/- 0.64 in NX, P < 0.001) and localized to damaged tubules. Kim-1 protein expression was markedly induced in NX+BSA (2.46 +/- 1.19 vs. 0.39 +/- 0.10% staining/field in NX, P < 0.001). Urinary Kim-1 was significantly elevated in NX+BSA (921 +/- 592 vs. 87 +/- 164 pg/ml in NX, P < 0.001) and correlated with tissue Kim-1 expression (r = 0.66, P =0.02). Kim-1 protein was found at the apical membrane of dilated nephrons. Kim-1 expression was limited to areas with inflammation (MO), fibrosis (alpha-smooth muscle actin), and tubular damage (osteopontin), and only occasionally with tubular dedifferentiation (vimentin). These results implicate involvement of Kim-1 in the pathogenesis of proteinuria-induced renal damage/repair. Urinary Kim-1 levels may serve as a marker of proteinuria-induced renal damage. PMID- 16467127 TI - Inhibition of p21 modifies the response of cortical proximal tubules to cisplatin in rats. AB - The purpose of this study was to evaluate whether upregulated p21, a cell cycle inhibitory protein, contributes to cisplatin (CDDP)-induced acute renal failure (ARF) and to acquired resistance to rechallenge injury with CDDP in rats. ARF was induced in rats by injection of CDDP (5 mg/kg) and rechallenge injury to CDDP by the same dose of CDDP 14 days after the first CDDP injection. Rats were treated with p21 antisense oligodeoxynucleotide (ODN) or its vehicle, p21 sense ODN, every 36 h from days 0 to 5 for single CDDP and from days 13 to 19 for rechallenge injury and killed at day 3, 5, 16, or 19. The uptake of FITC-labeled p21 antisense ODNs by cortical proximal tubule (PT) cells was much greater than by PT cells in the outer stripe of outer medulla (OSOM). Administration of antisense induced partial downregulation of p21 mRNA and protein levels in whole kidneys with single CDDP treatment and its rechallenge injury. Antisense significantly aggravated PT necrosis and decreased the number of p21-positive PT cells in the cortex but not in the OSOM in both CDDP-induced ARF and its rechallenge injury. However, antisense did not alter serum creatinine (Scr) and blood urea nitrogen (BUN) levels. Our findings suggested that p21 plays, at least in part, a cytoprotective role in cortical PTs exposed to CDDP, although this does not contribute to renal dysfunction when judged by Scr and BUN levels. Because antisense may not adequately be taken up and/or function in PTs in the OSOM, the role of p21 in PTs in the OSOM in CDDP-induced ARF remains to be clarified. PMID- 16467128 TI - Angiotensin II-induced calcium signaling in the afferent arteriole from rats with two-kidney, one-clip hypertension. AB - The aim of this study was to investigate ANG II-induced Ca2+ signaling in freshly isolated afferent arterioles (AA) from two-kidney, one-clip hypertensive (2K1C) rats, which have an elevated plasma and renal ANG II level, and different perfusion pressure and vascular tone in the clipped and nonclipped kidney. The Ca2+ responses in vessels from 2K1C and control rats were similar in all groups (P>0.1). The intracellular Ca2+ (Cai2+) response in the afferent arteriole after 10(-8) M ANG II stimulation was 0.57+/-0.10, 0.50+/-0.07, 0.48+/-0.04, and 0.36+/ 0.05 in the control, sham, nonclipped, and clipped kidney, respectively. These data were consistent with the finding of unchanged AT(1a)R mRNA levels in AAs from all groups. Although the absolute values were similar, the dose-response curves to ANG II were different. In the control, sham, and nonclipped kidney from 2K1C, the dose-response curve leveled off between 10(-8) and 10(-6) M ANG II. In the clipped kidney, the dose-response curve was linear, with a significantly increased response at 10(-6) M compared with 10(-8) M ANG II (P<0.05). Inhibition of cyclooxygenase-1 (COX-1) with indomethacin enhanced the ANG II response in the nonclipped (Delta0.30+/-0.09) and clipped (Delta0.30+/-0.09) kidneys from 2K1C (P<0.005), but not in control rats (Delta-0.02+/-0.11, P>0.8). Conclusively, the ANG II-induced Cai2+ response was reduced by COX-1-derived prostaglandins in 2K1C, in contrast to control animals, where the COX-1 inhibition had no effect. COX-2 inhibition with NS-398 did not increase the ANG II-mediated Cai2+ response in any of the groups. PMID- 16467129 TI - Automated method for the isolation of collecting ducts. AB - The structural and functional heterogeneity of the collecting duct present a tremendous experimental challenge requiring manual microdissection, which is time consuming, labor intensive, and not amenable to high throughput. To overcome these limitations, we developed a novel approach combining the use of transgenic mice expressing green fluorescent protein (GFP) in the collecting duct with large particle-based flow cytometry to isolate pure populations of tubular fragments from the whole collecting duct (CD), or inner medullary (IMCD), outer medullary (OMCD), or connecting segment/cortical collecting duct (CNT/CCD). Kidneys were enzymatically dispersed into tubular fragments and sorted based on tubular length and GFP intensity using large-particle-based flow cytometry or a complex object parametric analyzer and sorter (COPAS). A LIVE/DEAD assay demonstrates that the tubules were >90% viable. Tubules were collected as a function of fluorescent intensity and analyzed by epifluorescence and phase microscopy for count accuracy, GFP positivity, average tubule length, and time required to collect 100 tubules. Similarly, mRNA and protein from sorted tubules were analyzed for expression of tubule segment-specific genes using quantitative real-time RT-PCR and immunoblotting. The purity and yield of sorted tubules were related to sort stringency. Four to six replicates of 100 collecting ducts (9.68+/-0.44-14.5+/ 0.66 cm or 9.2+/-0.7 mg tubular protein) were routinely obtained from a single mouse in under 1 h. In conclusion, large-particle-based flow cytometry is fast, reproducible, and generates sufficient amounts of highly pure and viable collecting ducts from single or replicate animals for gene expression and proteomic analysis. PMID- 16467130 TI - Acute effect of high glucose on long-term cell growth: a role for transient glucose increase in proximal tubule cell injury. AB - Although chronic exposure of renal cells to high glucose has been shown to cause cell injury, the effect of acute exposure has not been elucidated. In this study, we demonstrate that acute (10 min) exposure of human proximal tubule epithelial cells (hPTEC) to high glucose (25 mM) induces a time-dependent dual effect consisting of an early proliferation and a late apoptosis. Acute exposure of hPTEC to high glucose induced a twofold increase in DNA synthesis and cell number at 12 h. However, after 36 h, a significant decrease in cell growth is observed, followed by apoptosis. On glucose treatment, both p42/p44 mitogen-activated protein (MAP) kinases and the downstream signaling intermediate NF-kappaB were phosphorylated and translocated to the nucleus. Pretreatment of cells with MAP kinase and NF-kappaB-specific inhibitors abolished glucose-induced proliferation. However, these inhibitors were ineffective in preventing glucose-induced apoptosis. Interestingly, conditioned medium from cells exposed to high-glucose concentrations inhibited proliferation and concomitantly induced apoptosis in normal cells, suggesting that the inhibitory effect of glucose occurs through secretion of a secondary factor(s). In parallel to apoptosis, we observed an increased production of reactive oxygen species (ROS). Pretreatment of cells with the antioxidant N-acetyl cysteine reversed glucose-mediated ROS production and apoptosis, suggesting that ROS is involved in apoptosis. Our study demonstrates for the first time that a single high-glucose exposure for 10 min alone is sufficient to elicit proliferation and apoptosis in hPTEC and suggests that episodes of transient increase in glucose may contribute to cell damage leading to epithelial cell dysfunction. PMID- 16467131 TI - TRPV4: a new target for the hypertension-related kinases WNK1 and WNK4. PMID- 16467132 TI - The human intestinal cytochrome P450 "pie". AB - Cytochromes P450 (P450s) 3A, 2C, and 1A2 constitute the major "pieces" of the human liver P450 "pie" and account, on average, for 40, 25, and 18%, respectively, of total immunoquantified P450s (J Pharmacol Exp Ther 270:414-423, 1994). The P450 profile in the human small intestine has not been fully characterized. Therefore, microsomes prepared from mucosal scrapings from the duodenal/jejunal portion of 31 human donor small intestines were analyzed by Western blot using selective P450 antibodies. P450s 3A4, 2C9, 2C19, and 2J2 were detected in all individuals and ranged from 8.8 to 150, 2.9 to 27, <0.6 to 3.9, and <0.2 to 3.1 pmol/mg, respectively. CYP2D6 was detected in 29 individuals and ranged from <0.2 to 1.4 pmol/mg. CYP3A5 was detected readily in 11 individuals, with a range (average) of 4.9 to 25 (16) pmol/mg that represented from 3 to 50% of total CYP3A (CYP3A4 + CYP3A5) content. CYP1A1 was detected readily in three individuals, with a range (average) of 3.6 to 7.7 (5.6) pmol/mg. P450s 1A2, 2A6, 2B6, 2C8, and 2E1 were not or only faintly detected. As anticipated, average CYP3A content (50 pmol/mg) was the highest. Excluding CYP1A1, the remaining enzymes had the following rank order: 2C9 > 2C19 > 2J2 > 2D6 (8.4, 1.1, 0.9, and 0.5 pmol/mg, respectively). Analysis of a pooled preparation of the 31 donor specimens substantiated these results. In summary, as in the liver, large interindividual variation exists in the expression levels of individual P450s. On average, CYP3A and CYP2C9 represents the major pieces of the intestinal P450 pie, accounting for 80 and 15%, respectively, of total immunoquantified P450s. PMID- 16467133 TI - An examination of the interplay between enterocyte-based metabolism and lymphatic drug transport in the rat. AB - The current study has examined whether drugs that are transported to the systemic circulation via the intestinal lymph (and therefore associate with lipoproteins within the enterocyte) are accessible to enterocyte-based metabolic processes. The impact of changes to the mass of lipid present within the enterocyte-based lymph lipid precursor pool (LLPP) on the extent of enterocyte-based drug metabolism has also been addressed. Low (5 mg oleic acid/h) or high [20 mg oleic acid/5.2 mg lyso-phosphatidylcholine/h] lipid dose formulations containing halofantrine (which is lymphatically transported and metabolized) or dichlorodiphenyltrichloroethane (DDT) (which is lymphatically transported and relatively metabolically inert) and radiolabeled oleic acid were infused into the duodenum of lymph duct-cannulated rats. After 5 h, drug and radiolabeled oleic acid were removed from the infusions, allowing calculation of the first order turnover rate constants describing drug and oleic acid transport from the LLPP into lymph from the washout profiles. In one group of animals, bolus doses of ketoconazole were also administered to inhibit cytochrome P450-based metabolism. The rate constant describing halofantrine transport from the LLPP into the lymph was lower than that of oleic acid, whereas these differences were abolished in the presence of ketoconazole. DDT and oleic acid exhibited similar turnover rate constants. The data therefore suggest that enterocyte-based metabolism removes halofantrine from the LLPP before transport into the lymph. Furthermore, enhancing the lymphatic transport of halofantrine by coadministration of larger quantities of lipid reduced the difference between the turnover rate constant for halofantrine and oleic acid and seemed to reduce the extent of enterocyte-based metabolism. PMID- 16467134 TI - Biotransformation of carbon-14-labeled muraglitazar in male mice: interspecies difference in metabolic pathways leading to unique metabolites. AB - Muraglitazar (Pargluva; Bristol-Myers Squibb), a dual alpha/gamma peroxisome proliferator-activated receptor activator, is under development for treatment of type 2 diabetes. This study describes the biotransformation profile of carbon-14 labeled muraglitazar in plasma, urine, feces, and bile samples from male CD-1 mice [intact and bile duct cannulation (BDC)] after single oral doses of 1 and 40 mg/kg. The major drug-related component circulating in mouse plasma was the parent compound for up to 4 h postdose. Similar to excretion profiles of muraglitazar in humans, monkeys, and rats, urinary excretion was the minor and fecal excretion via the biliary route was the major elimination pathway for muraglitazar in mice. The parent compound was a minor component in urine, bile, and feces, indicating that muraglitazar was extensively metabolized in mice. Major biotransformation pathways of muraglitazar in mice included taurine conjugate formation, acyl glucuronidation, hydroxylation, and O-dealkylation. In addition to those metabolites previously identified in humans, monkeys, and rats (M1-M21), several unique metabolites identified in mice included taurine conjugates (M24, M25, M26a,b,c, and M31), oxazole-ring-opened metabolites (M27 and M28), glutathione conjugates (M29a,b and M30), a dihydroxylated metabolite (M32), hydroxylated metabolites (M33 and M35), and a dehydrogenated metabolite (M34). The taurine conjugate of muraglitazar, M24, was a major metabolite in mice, accounting for 12 to 15% of the total dose in BDC mice or 7 to 12% of the total dose in intact mice. None of these taurine and glutathione conjugates were found in the bile samples of humans, monkeys, or rats. PMID- 16467135 TI - Duration of pleconaril effect on cytochrome P450 3A activity in healthy adults using the oral biomarker midazolam. AB - The objective of this study was to evaluate the duration of oral pleconaril (a picornavirus inhibitor) effect on intestinal and hepatic cytochrome P450 (P450) 3A activity as assessed by oral midazolam. Healthy adults received oral midazolam (0.075 mg/kg) on days 1 (baseline), 7, 9, 13, 20, 27, and 34. Oral pleconaril (400 mg) three times daily for 15 doses was administered on days 2 through 7. Blood samples were collected during each day of midazolam dosing to determine plasma midazolam concentrations. On days 5, 6, and 7, blood samples were collected to determine plasma pleconaril concentrations. Midazolam pharmacokinetics were determined by noncompartmental analyses, with bioequivalence assessed by least-squares geometric mean ratios (LS-GMR) and 90% confidence intervals (90% CI). Eighteen subjects completed the study. Midazolam C(max) (LS-GMR; 90% CI) decreased 24% on day 7 (0.76; 0.66-0.87). Midazolam oral clearance increased 53% on day 7 (1.53; 1.38-1.69). Midazolam oral clearance remained different on days 9 (1.38; 1.25-1.52) and 13 (1.19; 1.07-1.31) versus day 1. Midazolam volume of distribution (1.82; 1.57-2.11) and elimination half life (1.19; 1.03-1.38) were also different on day 7 in comparison with day 1. Oral pleconaril increased intestinal and hepatic CYP3A activity. The duration of increased CYP3A activity by pleconaril was at least 6 days (but no longer than 13 days) after pleconaril discontinuation. PMID- 16467136 TI - Pimecrolimus: absorption, distribution, metabolism, and excretion in healthy volunteers after a single oral dose and supplementary investigations in vitro. AB - The absorption and disposition of pimecrolimus, a calcineurin inhibitor developed for the treatment of inflammatory skin diseases, was investigated in four healthy volunteers after a single oral dose of 15 mg of [(3)H]pimecrolimus. Supplementary information was obtained from in vitro experiments. Pimecrolimus was rapidly absorbed. After t(max) (1-3 h), its blood concentrations fell quickly to 3% of C(max) at 24 h, followed by a slow terminal elimination phase (average t(1/2) 62 h). Radioactivity in blood decreased more slowly (8% of C(max) at 24 h). The tissue and blood cell distribution of pimecrolimus was high. The metabolism of pimecrolimus in vivo, which could be well reproduced in vitro (human liver microsomes), was highly complex and involved multiple oxidative O-demethylations and hydroxylations. In blood, pimecrolimus was the major radiolabeled component up to 24 h (49% of radioactivity area under the concentration-time curve(0-24) h), accompanied by a large number of minor metabolites. The average fecal excretion of radioactivity between 0 and 240 h amounted to 78% of dose and represented predominantly a complex mixture of metabolites. In urine, 0 to 240 h, only about 2.5% of the dose and no parent drug was excreted. Hence, pimecrolimus was eliminated almost exclusively by oxidative metabolism. The biotransformation of pimecrolimus was largely catalyzed by CYP3A4/5. Metabolite pools generated in vitro showed low activity in a calcineurin-dependent T-cell activation assay. Hence, metabolites do not seem to contribute significantly to the pharmacological activity of pimecrolimus. PMID- 16467137 TI - Crystal structure of obelin after Ca2+-triggered bioluminescence suggests neutral coelenteramide as the primary excited state. AB - The crystal structure at 1.93-A resolution is determined for the Ca2+-discharged obelin containing three bound calcium ions as well as the product of the bioluminescence reaction, coelenteramide. This finding extends the series of available spatial structures of the ligand-dependent conformations of the protein to four, the obelin itself, and those after the bioluminescence reaction with or without bound Ca2+ and/or coelenteramide. Among these structures, global conformational changes are small, typical of the class of "calcium signal modulators" within the EF-hand protein superfamily. Nevertheless, in the active site there are significant repositions of two residues. The His-175 imidazole ring flips becoming almost perpendicular to the original orientation corroborating the crucial importance of this residue for triggering bioluminescence. Tyr-138 hydrogen bonded to the coelenterazine N1-atom in unreacted obelin is moved away from the binding cavity after reaction. However, this Tyr is displaced by a water molecule from within the cavity, which now forms a hydrogen bond to the same atom, the amide N of coelenteramide. From this observation, a reaction scheme is proposed that would result in the neutral coelenteramide as the primary excited state product in photoprotein bioluminescence. From such a higher energy state it is now energetically feasible to account for the shorter wavelength bioluminescence spectra obtained from some photoprotein mutants or to populate the lower energy state of the phenolate anion to yield the blue bioluminescence ordinarily observed from native photoproteins. PMID- 16467138 TI - Effect of protein kinase A on accumulation of brefeldin A-inhibited guanine nucleotide-exchange protein 1 (BIG1) in HepG2 cell nuclei. AB - Brefeldin A-inhibited guanine nucleotide-exchange proteins, BIG1 and BIG2, are activators of ADP-ribosylation factor GTPases that are essential for regulating vesicular traffic among intracellular organelles. Biochemical analyses and immunofluorescence microscopy demonstrated BIG1 in nuclei as well as membranes and cytosol of serum-starved HepG2 cells. Within 20 min after addition of 8-Br cAMP, BIG1 accumulated in nuclei, and this effect was blocked by protein kinase A (PKA) inhibitors H-89 and PKI, suggesting a dependence on PKA-catalyzed phosphorylation. BIG2 localization was not altered by cAMP, nor did BIG2 small interfering RNA influence nuclear accumulation of BIG1 induced by cAMP. Mutant BIG1 (S883A) in which Ala replaced Ser-883, a putative PKA phosphorylation site, did not move to the nucleus with cAMP addition, whereas replacement with Asp (S883D) resulted in nuclear accumulation of BIG1 without or with cAMP exposure, consistent with the mechanistic importance of a negative charge at that site. Mutation (712KPK714) of the nuclear localization signal inhibited BIG1 accumulation in nuclei, and PKA-catalyzed phosphorylation of S883, although necessary, was not sufficient for nuclear accumulation, as shown by the double mutation S883D/nuclear localization signal. A role for microtubules in cAMP induced translocation of BIG1 is inferred from its inhibition by nocodazole. Thus, two more critical elements of BIG1 molecular structure were identified, as well as the potential function of microtubules in a novel PKA effect on BIG1 translocation. PMID- 16467139 TI - Direct structural observation of a molecular junction by high-energy x-ray reflectometry. AB - We report a direct angstrom resolution measurement of the structure of a molecular-size electronic junction comprising a single (or a double) layer of alkyl-thiol and alkyl-silane molecules at the buried interface between solid silicon and liquid mercury. The high-energy synchrotron x-ray measurements reveal densely packed layers comprising roughly interface-normal molecules. The monolayer's thickness is found to be 3-4 A larger than that of similar layers at the free surfaces of both mercury and silicon. The origins of this and the other unusual features detected are discussed in this article. Measurements of the bilayer junction with an applied potential did not show visible changes in the surface normal structure. PMID- 16467140 TI - Buffering of crucial functions by paleologous duplicated genes may contribute cyclicality to angiosperm genome duplication. AB - Genome duplication followed by massive gene loss has permanently shaped the genomes of many higher eukaryotes, particularly angiosperms. It has long been believed that a primary advantage of genome duplication is the opportunity for the evolution of genes with new functions by modification of duplicated genes. If so, then patterns of genetic diversity among strains within taxa might reveal footprints of selection that are consistent with this advantage. Contrary to classical predictions that duplicated genes may be relatively free to acquire unique functionality, we find among both Arabidopsis ecotypes and Oryza subspecies that SNPs encode less radical amino acid changes in genes for which there exists a duplicated copy at a "paleologous" locus than in "singleton" genes. Preferential retention of duplicated genes encoding long complex proteins and their unexpectedly slow divergence (perhaps because of homogenization) suggest that a primary advantage of retaining duplicated paleologs may be the buffering of crucial functions. Functional buffering and functional divergence may represent extremes in the spectrum of duplicated gene fates. Functional buffering may be especially important during "genomic turmoil" immediately after genome duplication but continues to act approximately 60 million years later, and its gradual deterioration may contribute cyclicality to genome duplication in some lineages. PMID- 16467141 TI - In search of rat stem Leydig cells: identification, isolation, and lineage specific development. AB - Leydig cells (LCs) are thought to differentiate from spindle-shaped precursor cells that exhibit some aspects of differentiated function, including 3beta hydroxysteroid dehydrogenase (3betaHSD) activity. The precursor cells ultimately derive from undifferentiated stem LCs (SLCs), which are postulated to be present in testes before the onset of precursor cell differentiation. We searched for cells in the neonatal rat testis with the abilities to: (i) proliferate and expand indefinitely in vitro (self renew); (ii) differentiate (i.e., 3betaHSD and ultimately synthesize testosterone); and (iii) when transplanted into host rat testes, colonize the interstitium and subsequently differentiate in vivo. At 1 week postpartum, spindle-shaped cells were seen in the testicular interstitium that differed from the precursor cells in that they were 3betaHSD-negative, luteinizing hormone (LH) receptor (LHR)-negative, and platelet-derived growth factor receptor alpha (PDGFR alpha)-positive. These cells were purified from the testes of 1-week-old rats. The cells contained proteins known to be involved in LC development, including GATA4, c-kit receptor, and leukemia inhibitory factor receptor. The putative SLCs expanded over the course of 6 months while remaining undifferentiated. When treated in media that contained thyroid hormone, insulin like growth factor I, and LH, 40% of the putative SLCs came to express 3betaHSD and to synthesize testosterone. When transplanted into host rat testes from which LCs had been eliminated, the putative SLCs colonized the interstitium and subsequently expressed 3betaHSD, demonstrating their ability to differentiate in vivo. We conclude that these cells are likely to be the sought-after SLCs. PMID- 16467142 TI - Flexible bilayers with spontaneous curvature lead to lamellar gels and spontaneous vesicles. AB - Mixtures of cetyltrimethylammonium tosylate (CTAT) and sodium dodecylbenzene sulfonate (SDBS) in water form a fluid lamellar phase at < or = 40 wt % water but surprisingly turn into viscous gels at higher water fractions. The gels are characterized by spherulite and other bilayer defects consistent with a low bending elasticity, kappa approximately k(B)T, and a nonzero spontaneous curvature. Caille analysis of the small-angle x-ray line shape confirms that for 7:3 wt:wt CTAT:SDBS bilayers at 50% water, kappa = 0.62 +/- 0.09 k(B)T and kappa = -0.9 +/- 0.2 k(B)T. For 13:7 wt:wt CTAT:SDBS bilayers, the measured bending elasticity decreases with increasing water dilution in good agreement with predictions based on renormalization theory, giving kappa(o) = 0.28 k(B)T. These results show that surfactant mixing is sufficient to make kappa approximately k(B)T, which promotes strong, Helfrich-type repulsion between bilayers that can dominate the van der Waals attraction. These are necessary conditions for spontaneous vesicles formed at even higher water fractions to be equilibrium structures. PMID- 16467143 TI - Directing electron transfer within Photosystem I by breaking H-bonds in the cofactor branches. AB - Photosystem I has two branches of cofactors down which light-driven electron transfer (ET) could potentially proceed, each consisting of a pair of chlorophylls (Chls) and a phylloquinone (PhQ). Forward ET from PhQ to the next ET cofactor (FX) is described by two kinetic components with decay times of approximately 20 and approximately 200 ns, which have been proposed to represent ET from PhQB and PhQA, respectively. Immediately preceding each quinone is a Chl (ec3), which receives a H-bond from a nearby tyrosine. To decrease the reduction potential of each of these Chls, and thus modify the relative yield of ET within the targeted branch, this H-bond was removed by conversion of each Tyr to Phe in the green alga Chlamydomonas reinhardtii. Together, transient optical absorption spectroscopy performed in vivo and transient electron paramagnetic resonance data from thylakoid membranes showed that the mutations affect the relative amplitudes, but not the lifetimes, of the two kinetic components representing ET from PhQ to F(X). The mutation near ec3A increases the fraction of the faster component at the expense of the slower component, with the opposite effect seen in the ec3B mutant. We interpret this result as a decrease in the relative use of the targeted branch. This finding suggests that in Photosystem I, unlike type II reaction centers, the relative efficiency of the two branches is extremely sensitive to the energetics of the embedded redox cofactors. PMID- 16467144 TI - ALADINI482S causes selective failure of nuclear protein import and hypersensitivity to oxidative stress in triple A syndrome. AB - Triple A syndrome is an autosomal recessive neuroendocrinological disease caused by mutations in a gene that encodes 546 amino acid residues. The encoded protein is the nucleoporin ALADIN, a component of nuclear pore complex (NPC). We identified a mutant ALADIN(I482S) that fails to target NPC and investigated the consequences of mistargeting using cultured fibroblasts (I482Sf) from a patient with triple A syndrome. ALADIN(I482S) affected a karyopherin-alpha/beta-mediated import pathway and decreased nuclear accumulations of aprataxin (APTX), a repair protein for DNA single-strand breaks (SSBs), and of DNA ligase I in I482Sf. This decrease was restored by wild-type ALADIN. ALADIN(I482S) had no effect on imports of M9/kap-beta2, BIB/kap-beta3, histone H1/importin 7, the ubiquitin conjugating enzyme UbcM2/importin 11, or the spliceosome protein U1A, indicating that ALADIN(I482S) selectively impaired transport of discrete import complexes through NPC. Cell survival assay showed hypersensitivity of I482Sf to l-buthionine-(S,R) sulfoximine (BSO), a glutathione-depleting agent. BSO decreased nuclear APTX and ligase I levels in I482Sf and normal control fibroblasts, but increased SSBs only in I482Sf. These observations implied that I482Sf are hypersensitive to BSO and no longer sufficiently repair SSBs. Consistent with this notion, I482Sf transfected with both APTX and ligase I had increased resistance to BSO, whereas I482Sf transfected with LacZ vector remained hypersensitive to BSO. We propose that oxidative stress aggravates nuclear import failure, which is already compromised in patient cells. Consequent DNA damage, beyond the limited capacity of DNA repair proteins, i.e., APTX and ligase I, may participate in triggering cell death. PMID- 16467145 TI - Quorum sensing and multidrug transporters in Escherichia coli. AB - Previously, we found that the quorum sensing transcription factor SdiA up regulates AcrAB. Others found that a 4-quinolone was a quorum-sensing signal in Pseudomonas aeruginosa. In Escherichia coli, there are at least three multidrug transporters (AcrAB/TolC, MdfA, and NorE) that exude fluoroquinolones. Here, we show that DeltaacrAB, tolC210, or DeltanorE mutants have the same growth rate as WT cells in exponential phase but grow to higher cell density in stationary phase. Overproduction of either pump caused cells to reach lower density. mdfA had no effect. Conditioned medium (CM) from cells overexpressing acrAB represses cell growth more than CM from WT cells. CM from pump mutant cells represses cell growth less than CM from WT cells. These results were not affected by the deletion of luxS, which synthesizes the quorum-sensing signal autoinducer 2 (AI 2). Expression of the rpoS gene encoding the stationary phase sigma factor is induced earlier in cells overexpressing acrAB and later in acrAB mutant cells. These results support a model in which a natural function of AcrAB/TolC and NorE is to export signals for cell-cell communication. Drugs exported by pumps may resemble communication molecules normally exuded. PMID- 16467146 TI - Ataxia telangiectasia mutated (Atm) is not required for telomerase-mediated elongation of short telomeres. AB - Telomerase-mediated telomere addition counteracts telomere shortening due to incomplete DNA replication. Short telomeres are the preferred substrate for telomere addition by telomerase; however, the mechanism by which telomerase recognizes short telomeres is unclear. In yeast, the Ataxia telangiectasia mutated (Atm) homolog, Tel1, is necessary for normal telomere length regulation likely by altering telomere structure, allowing telomerase recruitment to short telomeres. To examine the role of Atm in establishing preference for elongation of short telomeres in mice, we examined telomerase-mediated elongation of short dysfunctional telomeres in the presence or absence of Atm. Here we show that Atm is dispensable for elongation of short telomeres by telomerase, suggesting that telomerase recruitment in mammalian cells and in yeast may be regulated differently. PMID- 16467147 TI - Identification and characterization of a gonadotropin-inhibitory system in the brains of mammals. AB - Successful reproduction requires maintenance of the reproductive axis within fine operating limits through negative feedback actions of sex steroids. Despite the importance of this homeostatic process, our understanding of the neural loci, pathways, and neurochemicals responsible remain incomplete. Here, we reveal a neuropeptidergic pathway that directly links gonadal steroid actions to regulation of the reproductive system. An RFamide (Arg-Phe-NH2) peptide that inhibits gonadotropin release from quail pituitary was recently identified and named gonadotropin-inhibitory hormone (GnIH). Birds are known to have specialized adaptations associated with gonadotropin-releasing hormone (GnRH) regulation to optimize reproduction (e.g., encephalic photoreceptors), and the existence of a hypothalamic peptide inhibiting gonadotropins may or may not be another such specialization. To determine whether GnIH serves as a signaling pathway for sex steroid regulation of the reproductive axis, we used immunohistochemistry and in situ hybridization to characterize the distribution and functional role of this peptide in hamsters, rats, and mice. GnIH-immunoreactive (GnIH-ir) cell bodies are clustered in the mediobasal hypothalamus with pronounced projections and terminals throughout the CNS. In vivo GnIH administration rapidly inhibits luteinizing hormone secretion. Additionally, GnIH-ir neurons form close appositions with GnRH cells, suggesting a direct means of GnRH modulation. Finally, GnIH-ir cells express estrogen receptor-alpha and exhibit robust immediate early gene expression after gonadal hormone stimulation. Taken together, the distribution of GnIH efferents to neural sites regulating reproductive behavior and neuroendocrine secretions, expression of steroid receptors in GnIH-ir nuclei, and GnIH inhibition of luteinizing hormone secretion indicate the discovery of a system regulating the mammalian reproductive axis. PMID- 16467148 TI - Stem cell factor receptor induces progenitor and natural killer cell-mediated cardiac survival and repair after myocardial infarction. AB - Inappropriate cardiac remodeling and repair after myocardial infarction (MI) predisposes to heart failure. Studies have reported on the potential for lineage negative, steel factor positive (c-kit+) bone marrow-derived hematopoetic stem/progenitor cells (HSPCs) to repair damaged myocardium through neovascularization and myogenesis. However, the precise contribution of the c-kit signaling pathway to the cardiac repair process has yet to be determined. In this study, we sought to directly elucidate the mechanistic contributions of c-kit+ bone marrow-derived hematopoetic stem/progenitor cells in the maintenance and repair of damaged myocardium after MI. Using c-kit-deficient mice, we demonstrate the importance of c-kit signaling in preventing ventricular dilation and hypertrophy, and the maintenance of cardiac function after MI in c-kit-deficient mice. Furthermore, we show phenotypic rescue of cardiac repair after MI of c-kit deficient mice by bone marrow transplantation of wild-type HSPCs. The transplanted group also had reduced apoptosis and collagen deposition, along with an increase in neovascularization. To better understand the mechanisms underlying this phenotypic rescue, we investigated the gene expression pattern within the infarcted region by using microarray analysis. This analysis suggested activation of inflammatory pathways, specifically natural killer (NK) cell-mediated mobilization after MI in rescued hearts. This finding was confirmed by immunohistology and by using an NK blocker. Thus, our investigation revealed a previously uncharacterized role for c-kit signaling after infarction by mediating bone marrow-derived NK and angiogenic cell mobilization, which contributes to improved remodeling and cardiac function after MI. PMID- 16467149 TI - Guanosine 3',5'-bispyrophosphate coordinates global gene expression during glucose-lactose diauxie in Escherichia coli. AB - Guanosine 3',5'-bispyrophosphate (ppGpp), also known as "magic spot," has been shown to bind prokaryotic RNA polymerase to down-regulate ribosome production and increase transcription of amino acid biosynthesis genes during the stringent response to amino acid starvation. Because many environmental growth perturbations cause ppGpp to accumulate, we hypothesize ppGpp to have an overarching role in regulating the genetic program that coordinates transitions between logarithmic growth (feast) and growth arrest (famine). We used the classic glucose-lactose diauxie as an experimental system to investigate the temporal changes in transcription that accompany growth arrest and recovery in wild-type Escherichia coli and in mutants that lack RelA (ppGpp synthetase) and other global regulators, i.e., RpoS and Crp. In particular, diauxie was delayed in the relA mutant and was accompanied by a 15% decrease in the number of carbon sources used and a 3-fold overall decrease in the induction of RpoS and Crp regulon genes. Thus the data significantly expand the previously known role of ppGpp and support a model wherein the ppGpp-dependent redistribution of RNA polymerase across the genome is the driving force behind control of the stringent response, general stress response, and starvation-induced carbon scavenging. Our conceptual model of diauxie describes these global control circuits as dynamic, interconnected, and dependent upon ppGpp for the efficient temporal coordination of gene expression that programs the cell for transitions between feast and famine. PMID- 16467150 TI - Peroxisome proliferator-activated receptor delta promotes very low-density lipoprotein-derived fatty acid catabolism in the macrophage. AB - Significant attention has focused on the role of low-density lipoprotein (LDL) in the pathogenesis of atherosclerosis. However, recent advances have identified triglyceride-rich lipoproteins [e.g., very LDL (VLDL)] as independent risk predictors for this disease. We have previously demonstrated peroxisome proliferator-activated receptor (PPAR)delta, but not PPARgamma, is the major nuclear VLDL sensor in the macrophage, which is a crucial component of the atherosclerotic lesion. Here, we show that, in addition to beta-oxidation and energy dissipation, activation of PPARdelta by VLDL particles induces key genes involved in carnitine biosynthesis and lipid mobilization mediated by a recently identified TG lipase, transport secretion protein 2 (also named desnutrin, iPLA2zeta, and adipose triglyceride lipase), resulting in increased fatty acid catabolism. Unexpectedly, deletion of PPARdelta results in derepression of target gene expression, a phenotype similar to that of ligand activation, suggesting that unliganded PPARdelta suppresses fatty acid utilization through active repression, which is reversed upon ligand binding. This unique transcriptional mechanism assures a tight control of the homeostasis of VLDL-derived fatty acid and provides a therapeutic target for other lipid-related disorders, including dyslipidemia and diabetes, in addition to coronary artery disease. PMID- 16467151 TI - Comparative genomics provides evidence for close evolutionary relationships between the urotensin II and somatostatin gene families. AB - Although urotensin II (UII) and somatostatin 1 (SS1) exhibit some structural similarities, their precursors do not show any appreciable sequence identity and, thus, it is widely accepted that the UII and SS1 genes do not derive from a common ancestral gene. The recent characterization of novel isoforms of these two peptides, namely urotensin II-related peptide (URP) and somatostatin 2 (SS2)/cortistatin (CST), provides new opportunity to revisit the phylogenetic relationships of UII and SS1 using a comparative genomics approach. In the present study, by radiation hybrid mapping and in silico sequence analysis, we have determined the chromosomal localization of the genes encoding UII- and somatostatin-related peptides in several vertebrate species, including human, chicken, and zebrafish. In most of the species investigated, the UII and URP genes are closely linked to the SS2/CST and SS1 genes, respectively. We also found that the UII-SS2/CST locus and the URP/SS1 locus are paralogous. Taken together, these data indicate that the UII and URP genes, on the one hand, and the SS1 and SS2/CST genes, on the other hand, arose through a segmental duplication of two ancestral genes that were already physically linked to each other. Our results also suggest that these two genes arose themselves through a tandem duplication of a single ancestral gene. It thus appears that the genes encoding UII- and somatostatin-related peptides belong to the same superfamily. PMID- 16467152 TI - N-arachidonoyl L-serine, an endocannabinoid-like brain constituent with vasodilatory properties. AB - The endocannabinoid N-arachidonoyl ethanolamine (anandamide), found both in the CNS and in the periphery, plays a role in numerous physiological systems. One might expect that the chemically related N-arachidonoyl-L-serine (ARA-S) could also be formed alongside anandamide. We have now isolated ARA-S from bovine brain and elucidated its structure by comparison with synthetic ARA-S. Contrary to anandamide, ARA-S binds very weakly to cannabinoid CB1 and CB2 or vanilloid TRPV1 (transient receptor potential vanilloid 1) receptors. However, it produces endothelium-dependent vasodilation of rat isolated mesenteric arteries and abdominal aorta and stimulates phosphorylation of p44/42 mitogen-activated protein (MAP) kinase and protein kinase B/Akt in cultured endothelial cells. ARA S also suppresses LPS-induced formation of TNF-alpha in a murine macrophage cell line and in wild-type mice, as well as in mice deficient in CB1 or CB2 receptors. Many of these effects parallel those reported for abnormal cannabidiol (Abn-CBD), a synthetic agonist of a putative novel cannabinoid-type receptor. Hence, ARA-S may represent an endogenous agonist for this receptor. PMID- 16467154 TI - Profile of Don Helmberger. PMID- 16467153 TI - Prion protein is expressed on long-term repopulating hematopoietic stem cells and is important for their self-renewal. AB - Although the wild-type prion protein (PrP) is abundant and widely expressed in various types of tissues and cells, its physiological function(s) remain unknown, and PrP knockout mice do not exhibit overt and undisputed phenotypes. Here we showed that PrP is expressed on the surface of several bone marrow cell populations successively enriched in long-term (LT) hematopoietic stem cells (HSCs) using flow cytometry analysis. Affinity purification of the PrP-positive and -negative fractions from these populations, followed by competitive bone marrow reconstitution assays, shows that all LT HSCs express PrP. HSCs from PrP null bone marrow exhibited impaired self-renewal in serial transplantation of lethally irradiated mouse recipients both in the presence and absence of competitors. When treated with a cell cycle-specific myelotoxic agent, the animals reconstituted with PrP-null HSCs exhibit increased sensitivity to hematopoietic cell depletion. Ectopic expression of PrP in PrP-null bone marrow cells by retroviral infection rescued the defective hematopoietic engraftment during serial transplantation. Therefore, PrP is a marker for HSCs and supports their self-renewal. PMID- 16467155 TI - Unraveling the complexity of flux regulation: a new method demonstrated for nutrient starvation in Saccharomyces cerevisiae. AB - An important question is to what extent metabolic fluxes are regulated by gene expression or by metabolic regulation. There are two distinct aspects to this question: (i) the local regulation of the fluxes through the individual steps in the pathway and (ii) the influence of such local regulation on the pathway's flux. We developed regulation analysis so as to address the former aspect for all steps in a pathway. We demonstrate the method for the issue of how Saccharomyces cerevisiae regulates the fluxes through its individual glycolytic and fermentative enzymes when confronted with nutrient starvation. Regulation was dissected quantitatively into (i) changes in maximum enzyme activity (Vmax, called hierarchical regulation) and (ii) changes in the interaction of the enzyme with the rest of metabolism (called metabolic regulation). Within a single pathway, the regulation of the fluxes through individual steps varied from fully hierarchical to exclusively metabolic. Existing paradigms of flux regulation (such as single- and multisite modulation and exclusively metabolic regulation) were tested for a complete pathway and falsified for a major pathway in an important model organism. We propose a subtler mechanism of flux regulation, with different roles for different enzymes, i.e., "leader," "follower," or "conservative," the latter attempting to hold back the change in flux. This study makes this subtlety, so typical for biological systems, tractable experimentally and invites reformulation of the questions concerning the drives and constraints governing metabolic flux regulation. PMID- 16467156 TI - An essential role for the hematopoietic transcription factor Ikaros in hypothalamic-pituitary-mediated somatic growth. AB - Ikaros transcription factors play critical functions in the control of lymphohematopoiesis and immune regulation. Family members contain multiple zinc fingers that mediate DNA binding and homooligomerization or heterooligomerization. Ikaros is abundantly expressed in pituitary mammosomatotrophs, where it deacetylates histone 3 sites on the proximal growth hormone (GH) promoter to silence gene expression. Ikaros-null mice display stunted growth with reduced circulating levels of the GH target factor insulin like growth factor I (IGF-I). Ikaros-deficient mice have small anterior pituitary glands with a disproportionately reduced somatotroph population. Systemic administration of GH results in increased IGF-I levels and enhanced somatic growth. In contrast, reconstitution with WT lymphocytes was not sufficient to rescue the stunted growth phenotype of Ikaros-deficient mice. Ikaros was identified in mouse hypothalamic arcuate nuclei, where it colocalized with GH releasing hormone (GHRH); in contrast, Ikaros-null mice lack GHRH immunoreactivity in the hypothalamus. Overexpression of Ikaros enhanced GHRH promoter activity and induced endogenous GHRH gene expression. These findings unmask a wider role for Ikaros in the neuroendocrine system, highlighting a critical contribution to the development of the hypothalamic-pituitary somatotrophic axis. PMID- 16467157 TI - In situ analysis of nitrogen fixation and metabolic switching in unicellular thermophilic cyanobacteria inhabiting hot spring microbial mats. AB - Genome sequences of two Synechococcus ecotypes inhabiting the Octopus Spring microbial mat in Yellowstone National Park revealed the presence of all genes required for nitrogenase biosynthesis. We demonstrate that nif genes of the Synechococcus ecotypes are expressed in situ in a region of the mat that varies in temperature from 53.5 degrees C to 63.4 degrees C (average 60 degrees C); transcripts are only detected at the end of the day when the mat becomes anoxic. Nitrogenase activity in mat samples was also detected in the evening. Hitherto, N2 fixation in hot spring mats was attributed either to filamentous cyanobacteria (not present at >50 degrees C in these mats) or to heterotrophic bacteria. To explore how energy-generating processes of the Synechococcus ecotypes track natural light and O2 conditions, we evaluated accumulation of transcripts encoding proteins involved in photosynthesis, respiration, and fermentation. Transcripts from photosynthesis (cpcF, cpcE, psaB, and psbB) and respiration (coxA and cydA) genes declined in the evening. In contrast, transcripts encoding enzymes that may participate in fermentation fell into two categories; some (ldh, pdhB, ald, and ackA) decreased in the evening, whereas others (pflB, pflA, adhE, and acs) increased at the end of the day and remained high into the night. Energy required for N2 fixation during the night may be derived from fermentation pathways that become prominent as the mat becomes anoxic. In a broader context, our data suggest that there are critical regulatory switches in situ that are linked to the diel cycle and that these switches alter many metabolic processes within the microbial mat. PMID- 16467158 TI - Design of a mimic of nonamyloidogenic and bioactive human islet amyloid polypeptide (IAPP) as nanomolar affinity inhibitor of IAPP cytotoxic fibrillogenesis. AB - Protein aggregation into cytotoxic oligomers and fibrils in vivo is linked to cell degeneration and the pathogenesis of >25 uncurable diseases, whereas the high aggregation propensity and insolubility of several bioactive polypeptides and proteins in vitro prevent their therapeutic use. Aggregation of human islet amyloid polypeptide (IAPP) into pancreatic amyloid is strongly associated with the pathogenesis of type II diabetes. IAPP is a 37-residue polypeptide that acts as a neuroendocrine regulator of glucose homeostasis. However, IAPP misfolds and self-associates into cytotoxic aggregates and fibrils even at nanomolar concentrations. Because IAPP aggregation causes beta-cell death and prohibits therapeutic application of IAPP in diabetes, we pursued a minimalistic chemical design approach to generate a molecular mimic of a nonamyloidogenic and bioactive IAPP conformation that would still be able to associate with IAPP and thus inhibit its fibrillogenesis and cytotoxicity. We show that the double N methylated full length IAPP analog [(N-Me)G24, (N-Me)I26]-IAPP (IAPP-GI) is a highly soluble, nonamyloidogenic, and noncytotoxic IAPP molecular mimic and an IAPP receptor agonist. Moreover, IAPP-GI binds IAPP with low nanomolar affinity and completely blocks IAPP cytotoxic self-assembly and fibrillogenesis with activity in the low nanomolar concentration range. Importantly, IAPP-GI dissociates cytotoxic IAPP oligomers and fibrils and is able to reverse their cytotoxicity. Bifunctional soluble IAPP mimics that combine bioactivity with the ability to block and reverse IAPP cytotoxic self-assembly are promising candidates for the treatment of diabetes. Moreover, our amyloid disease inhibitor design concept may be applicable to other protein aggregation diseases. PMID- 16467159 TI - Monitoring the direct and indirect damage of DNA bases and polynucleotides by using time-resolved infrared spectroscopy. AB - The nucleotide 5'-dGMP and polynucleotide poly(dGdC).poly(dGdC) have been irradiated by using a 200-fs, 200-nm laser pulses and spectrally characterized by using time-resolved infrared spectroscopy. Under the experimental conditions, 200 nm excitation generates both electronic excited states and radical cations through photoionization; the former decay rapidly to vibrationally hot ground state. By using infrared signatures we have been able to follow these processes, and at time scales of >1 ns we observe an infrared marker band at 1,702 cm(-1) within both 5'-dGMP and the polynucleotide assigned to a photoionized product of guanine. This transient has also been reproduced through indirect chemistry through the reaction with photogenerated carbonate radical with 5'-dGMP. The ability to use time-resolved infrared spectroscopy in this way paves the way for developing solution-phase studies to investigate both direct and indirect radiation chemistry of DNA. PMID- 16467160 TI - Streptococcal modulation of cellular invasion via TGF-beta1 signaling. AB - Group A Streptococcus (GAS) and other bacterial pathogens are known to interact with integrins as an initial step in a complex pathway of bacterial ingestion by host cells. Efficient GAS invasion depends on the interaction of bound fibronectin (Fn) with integrins and activation of integrin signaling. TGF-beta1 regulates expression of integrins, Fn, and other extracellular matrix proteins, and positively controls the integrin signaling pathway. Therefore, we postulated that TGF-beta1 levels could influence streptococcal invasion of mammalian cells. Pretreatment of HEp-2 cells with TGF-beta1 increased their capacity to ingest GAS when the bacteria expressed fibronectin-binding proteins (M1 or PrtF1). Western blots revealed significant induction of alpha5 integrin and Fn expression by HEp 2 cells in response to TGF-beta1. Increased ingestion of streptococci by these cells was blocked by a specific inhibitor of the TGF-beta1 receptor I and antibodies directed against alpha5 integrin and Fn, indicating that increased invasion depends on TGF-beta1 up-regulation of both the alpha5 integrin and Fn. The capacity of TGF-beta1 to up-regulate integrin expression and intracellular invasion by GAS was reproduced in primary human tonsil fibroblasts, which could be a source of TGF-beta1 in chronically infected tonsils. The relationship between TGF-beta1 and GAS invasion was strengthened by the observation that TGF beta1 production was stimulated in GAS-infected primary human tonsil fibroblasts. These findings suggest a mechanism by which GAS induce a cascade of changes in mammalian tissue leading to elevated expression of the alpha5beta1 receptor, enhanced invasion, and increased opportunity for survival and persistence in their human host. PMID- 16467161 TI - Rectal adenocarcinoma with oncocytic features: possible relationship with preoperative chemoradiotherapy. AB - BACKGROUND: The introduction of preoperative chemoradiation into the treatment protocol of rectal adenocarcinomas has affected the microscopical morphology in subsequent resection specimens. The constellation of histopathological changes is varied and well documented. AIM: To describe oncocytic change in rectal cancers that have been treated with chemoradiation before surgery. METHODS: 7 of 54 patients with rectal cancer were identified with a history of chemoradiation, specifically directed to the rectal tumours in fractions of 4500-5000 cGy of radiation and 5-fluorouracil. The rectal tumours in five of these seven patients were composed of oncocytes that constituted 30-80% of the cancers. The patients were three men and two women aged 65-73 years, all with T3 N0 tumours. The intervals between chemoradiation and resection varied from 3 to 12 weeks. RESULTS: The tumour cells conformed to oncocytes morphologically (large size with abundant, granular eosinophilic cytoplasm, vesicular nuclei and prominent acidophilic nucleoli), immunohistochemically (positive for carcinoembryonic antigen, cytokeratin 20 and caudal type homeo box transcription factor 2, but negative for both chromogranin and synaptophysin) and ultrastructurally (large cells showing tight junctions, cytoplasmic engorgement by mitochondria and absence of neurosecretory granules). CONCLUSIONS: The changes in these cells differ from those described previously in endocrine cells encountered in pretreated rectal cancers. Oncocytic change in this particular clinical context occurs as a reflection of cytotoxic damage or cellular hypoxia induced by chemoradiation resulting in degeneration of the cell and the oncocytic phenotype. Oncocytic change may be an under-recognised histopathological change in rectal cancers receiving preoperative chemoradiation. PMID- 16467162 TI - Post-traumatic fibro-osseous lesion of the ribs: a relatively under-recognised entity. AB - AIM: To report the description of a rare benign osseous lesion affecting the ribs entitled post-traumatic fibro-osseous lesion (PTFOL). METHODS: Seven cases of PTFOLs were retrieved from the archives of the University Hospital of Lille. Histological slides were reviewed and lesions were classified according to the histological patterns described by McDermott et al. Clinical and follow-up data were obtained from the patients' charts. RESULTS: PTFOLs occurred principally in men (mean age 31.8 years) with a known or suspected previous chest injury for four of them. No previous cancer was noted. Radiologically, PTFOLs readily presented as an isolated expansive lucency with a sclerotic rim located on the last five ribs. A constant increased uptake of radionucleotide was noted on bone scan. Microscopically, two fibro-osseous, four xanthomatous and one mixed pattern were individualised. Lesions were characterised by a network of anastomosing bone trabeculae without osteoblast lining within a fibrous stroma. A zonal maturation from woven to peripheral lamellar bone was characterised. Central sheets of lipid laden histiocytes were conspicuous in the xanthomatous type. For each patient, clinical follow-up was excellent, without any recurrence. CONCLUSIONS: This is the second largest series of PTFOLs, which is considered to be a dysplastic healing process after trauma. It may be symptomatic or shown by imaging studies realised for unrelated reason. PTFOL is regularly misdiagnosed with other more common lesions of the ribs, such as fibro-osseous dysplasia, osteoma osteoid and benign fibrous histiocytoma. Its recognition is of importance because no follow up is needed after resection. PMID- 16467163 TI - Bacteraemia caused by Anaerotruncus colihominis and emended description of the species. AB - BACKGROUND: Anaerotruncus colihomonis is a newly described bacterial genus and species isolated from the stool specimens of children. Its clinical significance, however, is unknown. AIMS: To describe a case of A colihominis bacteraemia identified by 16S ribosomal RNA (rRNA) gene sequencing and provide an emended description of the species. METHODS: An unidentified anaerobic bacillus (strain HKU19) that stains Gram negative was subjected to characterisation by 16S rRNA gene sequencing, G+C content determination and electron microscopy. RESULTS: Strain HKU19 was isolated from the blood culture of a 78-year-old woman with nosocomial bacteraemia. It was found to be an anaerobic, non-motile, pleomorphic, thin bacillus that stains Gram negative. It produces Indole and utilises glucose and mannose. Identifying the strain to the species level was not possible by conventional phenotypic tests and commercial identification systems. The G+C content of strain HKU19 was found to be 53.43 mol%. A similarity of 99.3% nucleotide identities was found between the 16S rRNA gene sequence of strain HKU19 and that of A colihominis WAL 14 565(T), which was isolated from a human faecal specimen. In contrast with the original description of A colihominis, HKU19 was found to produce occasional oval, terminal spores, although the other phenotypic characteristics matched. Spores were also occasionally observed when the two previously reported strains were re-examined. CONCLUSIONS: Although the source of the bacteraemia in the patient cannot be determined, this report suggests that A colihominis is of clinical significance. Spore formation is proposed as an emended description of A colihominis. PMID- 16467164 TI - Loss of human leucocyte antigen class I and gain of class II expression are early events in carcinogenesis: clues from a study of Barrett's oesophagus. AB - BACKGROUND: Human leucocyte antigen (HLA) expression is altered in oesophageal carcinomas compared with normal tissue. It is unclear, however, whether this phenotype precedes malignant transformation or results as a consequence of it. AIM: To investigate HLA class I and II expression in Barrett's oesophagus and normal squamous oesophageal tissue. METHODS: Asian patients with Barrett's oesophagus (n = 64) and a control group (n = 60) with a normal oesophagus but without reflux symptoms were recruited using endoscopic and histopathological criteria. Tissue samples were stained with monoclonal antibodies specific for HLA ABC, HLA-DR alpha chain or HLA-DP/DQ/DR, and scored semiquantitatively. The results of immunohistochemical staining were correlated with clinical and histopathological characteristics of patients. RESULTS: Marked expression of HLA ABC was observed in 50% of Barrett's oesophagus sections as compared with 68.3% of controls (p = 0.038). HLA-DR staining was seen in 51.6% of Barrett's oesophagus samples versus 11.7% of controls (p<0.001). Expression of HLA-DP/DQ/DR was evident in 73.4% of oesophageal intestinal metaplasia tissue as opposed to 18.3% of controls (p<0.001). Importantly, a total loss of HLA-ABC and a concomitant gain of HLA-DP/DQ/DR expression were seen in 37.5% of patients with Barrett's oesophagus but in none of the controls (p<0.001). Interestingly, this phenotype was associated positively with dysplasia (adjusted p, p* = 0.031) but negatively with non-steroidal anti-inflammatory drug use (p* = 0.004). CONCLUSIONS: HLA class I expression is down regulated and class II expression is up regulated in Barrett's oesophagus. As these changes predate malignant transformation, altered major histocompatibility complex expression may be a key event in disease progression, possibly in facilitating evasion from immune surveillance. PMID- 16467165 TI - Oncocytic change in pleomorphic adenoma: molecular evidence in support of an origin in neoplastic cells. AB - BACKGROUND: Cells with oncocytic change (OC) are a common finding in salivary glands (SGs) and in SG tumours. When found within pleomorphic adenomas (PAs), cells with OC may be perceived as evidence of malignancy, and lead to a misdiagnosis of carcinoma ex pleomorphic adenoma (CaExPa). AIM: To describe a case of PA with atypical OC, resembling a CaExPa. A genomewide molecular analysis was carried out to compare the molecular genetic features of the two components and to determine whether the oncocytic cells originated from PA cells, entrapped normal cells, or whether these cells constitute an independent tumour. MATERIALS AND METHODS: Representative blocks were immunohistochemically analysed with antibodies raised against cytokeratin (Ck) 5/6, Ck8/18, Ck14, vimentin, p63, alpha-smooth muscle actin (ASMA), S100 protein, anti-mitochondria antibody, beta catenin, HER2, Ki67, p53 and epidermal growth factor receptor. Typical areas of PA and OC were microdissected and subjected to microarray-based comparative genomic hybridisation (aCGH). Chromogenic in situ hybridisation (CISH) was performed with in-house generated probes to validate the aCGH findings. RESULTS: PA cells showed the typical immunohistochemical profile, including positivity for Ck5/6, Ck8/18, Ck14, vimentin, ASMA, S100 protein, p63, epidermal growth factor receptor and beta-catenin, whereas oncocytic cells showed a luminal phenotype, expression of anti-mitochondria antibody and reduced beta-catenin staining. Both components showed low proliferation rates and lacked p53 reactivity. aCGH revealed a similar amplification in both components, mapping to 12q13.3-q21.1, which was further validated by CISH. No HER2 gene amplification or overexpression was observed. The foci of oncocytic metaplasia showed an additional low-level gain of 6p25.2-p21.31. CONCLUSION: The present data demonstrate that the bizarre atypical cells of the present case show evidence of clonality but no features of malignancy. In addition, owing to the presence of a similar genome amplification pattern in both components, it is proposed that at least in some cases, OC may originate from PA cells. PMID- 16467166 TI - Immunocytochemistry of p16INK4a in liquid-based cervicovaginal specimens with modified Papanicolaou counterstaining. AB - AIM: To evaluate the feasibility and value of a modified Papanicolaou counterstain for p16(INK4a) immunostaining in liquid-based cervicovaginal samples. METHODS: Immunocytochemical analyses were carried out with p16(INK4a) and modified Papanicolaou counterstain on 81 liquid-based samples, including 23 of within normal limits (WNL), 6 of low-grade squamous intraepithelial lesion (LSIL), 20 of high-grade squamous intraepithelial lesion (HSIL), 16 of atypical squamous cells of undetermined significance (ASC-US) and 16 of atypical squamous cells, high-grade lesion cannot be excluded (ASC-H). Results were compared with histological or cytological follow-up. For comparison, samples from 29 more cases (10 of LSIL, 10 of ASC-H and 9 of HSIL) were immunostained with p16(INK4a) and conventionally counterstained with haematoxylin. The intensity of immunostaining in cases of squamous intraepithelial lesion (SIL) was assessed using a 0-3 scoring system. Interobserver agreement was calculated by kappa statistics. RESULTS: Expression of p16(INK4a) was detected in 3 of 23 cases of WNL, 4 of 6 cases of LSIL, all cases of HSIL, 5 of 16 cases of ASC-US and 13 of 16 cases of ASC-H. Excluding two cases with no residual dysplastic cells in the immunocytochemistry, all cases of cervical intraepithelial neoplasia (CIN)2 or CIN3 at follow-up expressed p16(INK4a) and none of the p16(INK4a)-negative cases showed a high-grade lesion at follow-up. No evident differences in pattern or intensity of p16(INK4a) expression were observed between the specimens of the study and control groups. Interobserver agreement was significantly better in the study group than in the group with conventional immunostaining (combined kappa 0.773 v 0.549; p<0.05), and still better, albeit statistically not significant, than with conventional immunostaining and cervical smear test together (combined kappa 0.773 v 0.642). CONCLUSION: Immunocytochemistry with p16(INK4a) and modified Papanicolaou counterstain may add to the cervicovaginal cytology the full potentiality of p16(INK4a) without the need of a further slide and the risk of loss of dysplastic cells, yet maintaining the typical morphological features of the smear test. PMID- 16467167 TI - Metaplastic carcinoma of the breast: a clinicopathological review. AB - BACKGROUND: Mammary metaplastic carcinoma encompasses epithelial-only carcinoma (high-grade adenosquamous carcinoma or pure squamous cell carcinoma), biphasic epithelial and sarcomatoid carcinoma and monophasic spindle cell carcinoma. AIM: To evaluate the clinicopathological features of a large series of 34 metaplastic carcinomas. METHODS: 10 epithelial-only, 14 biphasic and 10 monophasic metaplastic carcinomas were assessed for nuclear grade, hormone receptor status, HER2/neu (cerbB2) oncogene expression, Ki-67 and p53, lymph node status and recurrence on follow-up. RESULTS: Intermediate to high nuclear grade were assessed in most (33/34) tumours. Oestrogen and progesterone receptors were negative in 8 of 10 epithelial-only, all 14 biphasic, and 9 of 10 monophasic tumours, cerbB2 was negative in 7 of 10 epithelial-only, all 14 biphasic and 8 of 10 monophasic tumours. Ki-67 was found to be positive in 6 of 10 epithelial-only, 6 of 14 biphasic, and 7 of 10 monophasic tumours, whereas p53 was positive in 6 of 10 epithelial-only, 7 of 14 biphasic, and 8 of 10 monophasic tumours. Lymph node metastases were seen in 7 of 7 epithelial-only, 7 of 11 biphasic, and 3 of 7 monophasic tumours. Recurrences were seen in 4 of 7 epithelial-only, 8 of 9 biphasic, and 4 of 9 monophasic tumours. CONCLUSIONS: All three subtypes of metaplastic carcinoma are known to behave aggressively, and should be differentiated from the low-grade fibromatosis-like metaplastic carcinoma, which does not metastasize. Oncological treatment options may be limited by the frequently negative status of hormonal receptor and cerbB2. PMID- 16467168 TI - Correlation of clinicopathological features with immunohistochemical expression of cell cycle regulatory proteins p16 and retinoblastoma: distinct association with keratinisation and differentiation in oral cavity squamous cell carcinoma. AB - BACKGROUND: The p16 and retinoblastoma (Rb) gene products are part of the retinoblastoma pathway controlling the G1-S transition of the cell cycle. Few studies on the expression of p16 and retinoblastoma proteins in oral cavity squamous carcinomas have been conducted. AIM: To correlate the expression of p16 and retinoblastoma proteins to clinicopathological characteristics in these tumours. METHODS: 45 patients with resected oral cavity squamous carcinoma were selected, for whom this was the initial treatment and who were followed up for 5 years or until death. Immunohistochemical stains with antibodies to the Rb and p16 gene products were carried out on paraffin wax-embedded tissue. Data on clinicopathological features such as tumour differentiation, nodal status, stage and survival outcome were collected. RESULTS: Retinoblastoma expression was seen in 39 of 45 (87%) patients and p16 expression in 6 of 45 (13%) patients. A significant inverse correlation was observed between retinoblastoma and p16 expression as nearly all retinoblastoma negative cases were p16 positive, and vice versa. When examined for clinicopathological correlates, it was found that all 39 tumours that expressed retinoblastoma displayed marked keratinisation and were of low-moderate histological grade. Conversely, five of the six tumours that expressed p16 were found to be poorly differentiated, with minimal keratin expression. CONCLUSIONS: Salient relationships were seen between expression of retinoblastoma and p16 and keratinisation. A marked loss of keratin production was evident in the tumours that expressed p16. Tumours expressing retinoblastoma were seen to exhibit more widespread keratinisation. In addition, an inverse staining pattern was found for retinoblastoma and p16 as retinoblastoma expressing tumours were nearly universally p16 negative and vice versa. No correlation of expression of either p16 or retinoblastoma was found with survival or stage. A link between the histologically observable morphology and expression of cell cycle regulatory protein with the expression of p16 and retinoblastoma has been suggested with keratinisation and differentiation of status. PMID- 16467169 TI - Cyclooxygenase-2 expression correlates with poor prognosis in pancreatic cancer. AB - BACKGROUND: Cyclooxygenase-2 (COX-2) overexpression is related to poor outcome in several cancers. COX-2 is upregulated in 42-90% of pancreatic ductal adenocarcinomas and is a potential target for chemotherapy. Earlier studies have not shown the expression of COX-2 to be a prognostic factor in pancreatic cancer. OBJECTIVE: To evaluate the prognostic value of COX-2 in a series of patients with pancreatic adenocarcinoma. METHODS: 128 patients operated on for pancreatic adenocarcinoma at Helsinki University Central Hospital between 1974 and 1998 provided sections from primary tumours which were immunohistochemically stained with a COX-2-antihuman monoclonal antibody. RESULTS: Cytoplasmic COX-2 reactivity (>5%) occurred in 46 specimens (36%), correlating neither with age, sex, stage, size, tumour stage, nodal metastases, nor grade. Lack of COX-2 expression correlated with distant metastases (p = 0.026). In univariate survival analysis, COX-2 expression (p = 0.0114), stage (p = 0.0002), grade (p = 0.0001), and age (p = 0.042) had prognostic significance. One, two, and five year survival rates were 51%, 32%, and 8% in the COX-2 negative groups compared with 34%, 5%, and 5% in the COX-2 positive groups (p = 0.011). Prognostic significance was especially high for patients operated on with curative intent (p = 0.004). In multivariate analysis, COX-2 was an independent prognostic factor (hazard ratio = 1.6 (95% confidence interval, 1.1 to 2.3)). CONCLUSIONS: Expression of COX-2 was associated with poor outcome from pancreatic ductal adenocarcinoma and was independent of tumour stage, grade, or age in multivariate analysis. PMID- 16467170 TI - Nuclear beta-catenin and Ki-67 expression in choriocarcinoma and its pre malignant form. AB - OBJECTIVE: To study the expression of nuclear beta-catenin and Ki-67 in patients with normal gestation products (NGP), complete hydatidiform moles (CHM), and choriocarcinoma to elucidate their roles in carcinogenesis and their interrelations. METHODS: Expression of nuclear beta-catenin and Ki-67 was studied by immunohistochemistry using paraffin embedded blocks. Sixty NGP, 60 CHM, and 10 choriocarcinomas were analysed. In addition, approximately 400 trophoblasts each in 40 NGP, 40 CHM, and 10 choriocarcinomas from the same batch of samples were microdissected for quantitative reverse transcriptase polymerase chain reaction (Q-RT-PCR) to compare beta-catenin mRNA concentration among them. RESULTS: In the chorionic villi of NGP, beta-catenin was consistently expressed in the nuclei of cytotrophoblasts but not syncytiotrophoblasts. Nuclear beta-catenin expression was comparatively reduced in CHM trophoblasts and was absent in choriocarcinoma. By contrast, Ki-67 expression was increased from cytotrophoblasts but not in syncytiotrophoblasts in the chorionic villi of NGP to CHM trophoblasts and choriocarcinoma. Using Q-RT-PCR, beta-catenin mRNA was detected in 10 NGP, 13 CHM, and three choriocarcinoma specimens, with median copy numbers of 43,230, 18,229, and 17,334 per 400 trophoblasts, respectively. A housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA was detected as a control in the NGP, CHM, and choriocarcinoma specimens, with median copy numbers of 51,300, 54,270, and 97,150 per 400 trophoblasts, respectively. Thus median beta catenin mRNA values after normalisation were 0.85 in NGP (n = 10), 0.31 in CHM (n = 13), and 0.16 in choriocarcinoma (n = 3). CONCLUSIONS: Decreased nuclear beta catenin expression and increased Ki-67 expression may be involved in choriocarcinoma carcinogenesis. The findings also suggest that nuclear beta catenin may play a role in trophoblast differentiation during normal placental development. PMID- 16467171 TI - The roles of specific genes implicated as circulating factors involved in normal and disordered phosphate homeostasis: frizzled related protein-4, matrix extracellular phosphoglycoprotein, and fibroblast growth factor 23. AB - Normal serum phosphate (Pi) concentrations are relatively tightly controlled by endocrine mediators of Pi balance. Recent data involving several disorders of Pi homeostasis have shed new light on the regulation of serum Pi balance. It has been hypothesized that circulating phosphaturic factors, or phosphatonins, exist that, when present at high serum concentrations, directly act on the kidney to induce renal Pi wasting. This review will focus upon recently discovered factors that are overexpressed in tumors associated with tumor-induced osteomalacia and have reported activity consistent with effecting Pi balance in vivo. Currently, the best-characterized group of phosphatonin-like polypeptides includes secreted frizzled related protein-4, matrix extracellular phosphoglycoprotein, and fibroblast growth factor-23. Our understanding of these factors will, in the short term, aid us in understanding normal Pi balance and, in the future, help to design novel therapeutic strategies for disorders of Pi handling. PMID- 16467174 TI - Small particles with big effects. PMID- 16467173 TI - Older people's views of advice about falls prevention: a qualitative study. AB - The aim of this study was to gain an understanding of older people's perceptions of falls prevention advice, and how best to design communications that will encourage older people to take action to prevent falls. Focus groups and interviews were carried out with 66 people aged 61-94 years recruited from a variety of settings, using falls prevention messages to stimulate discussion. Thematic analysis revealed that participants interpreted 'falls prevention' principally as meaning hazard reduction, use of aids and restriction of activity. Only one participant was aware that falls risk could be reduced by carrying out exercises to improve strength and balance. Falls prevention advice was typically regarded as useful in principle but not personally relevant or appropriate. Advice about falling was often depicted as common sense, only necessary for older or more disabled individuals, and potentially patronizing and distressing. Our findings suggest that older people do not reject falls prevention advice because of ignorance of their risk of falling, but because they see it as a potential threat to their identity and autonomy. Messages that focus on the positive benefits of improving balance may be more acceptable and effective than advice on falls prevention. PMID- 16467175 TI - Exhaled breath condensate pH: reflecting acidification of the airway at all levels. PMID- 16467176 TI - The highs and lows of intensive insulin therapy. PMID- 16467177 TI - Observational studies of inhaled corticosteroids in chronic obstructive pulmonary disease: misconstrued immortal time bias. PMID- 16467178 TI - Azithromycin in bronchiolitis obliterans: is the evidence strong enough? PMID- 16467179 TI - Lung transplantation: opportunities for research and clinical advancement. PMID- 16467180 TI - Profiling of the tetraspanin web of human colon cancer cells. AB - Tetraspanins are integral membrane proteins involved in a variety of physiological and pathological processes. In cancer, clinical and experimental studies have reported a link between tetraspanin expression levels and metastasis. Tetraspanins play a role as organizers of multimolecular complexes in the plasma membrane. Indeed each tetraspanin associates specifically with one or a few other membrane proteins forming primary complexes. Thus, tetraspanin tetraspanin associations lead to a molecular network of interactions, the "tetraspanin web." We performed a proteomic characterization of the tetraspanin web using a model of human colon cancer consisting of three cell lines derived from the primary tumor and two metastases (hepatic and peritoneal) from the same patient. The tetraspanin complexes were isolated after immunoaffinity purification using monoclonal antibodies directed against the tetraspanin CD9, and the associated proteins were separated by SDS-PAGE and identified by mass spectrometry using LC-MS/MS. This allowed the identification of 32 proteins including adhesion molecules (integrins, proteins with Ig domains, CD44, and epithelial cell adhesion molecule) (EpCAM), membrane proteases (ADAM10, TADG-15, and CD26/dipeptidyl peptidase IV), and signaling proteins (heterotrimeric G proteins). Importantly some components were differentially detected in the tetraspanin web of the three cell lines: the laminin receptor Lutheran/B-cell adhesion molecule (Lu/B-CAM) was expressed only on the primary tumor cells, whereas CD26/dipeptidyl peptidase IV and tetraspanin Co-029 were observed only on metastatic cells. Concerning Co-029, immunohistofluorescence showed a high expression of Co-029 on epithelial cells in normal colon and a lower expression in tumors, whereas heterogeneity in terms of expression level was observed on metastasis. Finally we demonstrated that epithelial cell adhesion molecule and CD9 form a new primary complex in the tetraspanin web. PMID- 16467182 TI - The rheumatoid wrist. AB - Wrist involvement is common in patients with rheumatoid arthritis. Individual patient assessment is important in determining functional deficits and treatment goals. Patients with persistent disease despite aggressive medical management are candidates for surgery. Soft-tissue procedures offer good symptomatic relief and functional improvement in the short term. Extensor and flexor tendons may rupture because of synovial infiltration and bony irritation. When rupture occurs, direct repair usually is not possible. However, when joints that are motored by the ruptured tendon are still functional, tendon transfer or grafting may be considered. Because of the progressive nature of the disease, dislocation and end stage arthritis often require stabilization with bony procedures. The distal radioulnar joint is usually affected first and is commonly treated with either the Darrach or the Sauve-Kapandji procedure. Partial wrist fusion offers a compromise between achieving stability of the affected radiocarpal joint and maintaining motion at the midcarpal joint. For pancarpal arthritis, total wrist fusion offers reliable pain relief at the cost of motion. Total wrist arthroplasty is an alternative that preserves motion; however, the outcomes of total wrist replacement are still being evaluated. PMID- 16467183 TI - Subaxial cervical spine trauma. AB - Subaxial cervical spine injuries are common, ranging in severity from minor ligamentous strain or spinous process fracture to complete fracture-dislocation with bone and ligament failure, resulting in severe spinal cord injury. Understanding the epidemiology, anatomy, biomechanics, and classification of subaxial cervical spine injuries is important. Emergent management of such injuries is based on obtaining an accurate clinical history, careful physical examination, and organized radiographic evaluation. Attaining a unified approach to the wide spectrum of subaxial cervical injuries is difficult. In addition, controversy exists regarding the safety of closed reduction in certain injury patterns and the administration of methylprednisolone for acute spinal cord injury. Definitive management (surgical or nonsurgical) is based on the assessment of the mechanical instability of the injury, the presence or absence of neurologic impairment, and various patient factors that may influence outcome. Several complications, including the deterioration of neurologic status, may occur with either surgical or nonsurgical management, but the most frequent mistake made is missing the injury on initial evaluation. PMID- 16467184 TI - Osteochondritis dissecans of the knee. AB - Osteochondritis dissecans is a condition of the joints that appears to affect subchondral bone primarily, with secondary effects on articular cartilage. With progression, this pathology may present clinically with symptoms related to the integrity of the articular cartilage. Early signs, associated with intact cartilage, may be related to a softening phenomenon and alteration in the mechanical properties of cartilage. Later stages, because of the lack of underlying support of the cartilage, can present with signs of articular cartilage separation, cartilage flaps, loose bodies, inflammatory synovitis, persistent or intermittent joint effusion, and, in severe cases, secondary joint degeneration. Selecting and recommending a surgical intervention require balancing application of nonsurgical interventions with assessment of the degree of articular cartilage stability and the potential for spontaneous recovery. PMID- 16467185 TI - Early onset idiopathic scoliosis. AB - Children with early onset scoliosis typically present before age 5 years. Radiographic criteria help to distinguish progressive cases from those that will spontaneously resolve. Severe cardiopulmonary problems may occur in untreated progressive cases. A comprehensive evaluation should be performed to identify commonly associated conditions, such as plagiocephaly, congenital heart disease, inguinal hernia, and hip dysplasia. For curves >20 degrees , magnetic resonance imaging of the neural axis is indicated to rule out occult central nervous system lesions. Surgical management should be considered when nonsurgical measures, including bracing and casting, fail to arrest progression. Surgical methods continue to evolve and are primarily directed at obtaining and maintaining curve correction while simultaneously preserving spinal and trunk growth. PMID- 16467186 TI - Hardware removal: indications and expectations. AB - Although hardware removal is commonly done, it should not be considered a routine procedure. The decision to remove hardware has significant economic implications, including the costs of the procedure as well as possible work time lost for postoperative recovery. The clinical indications for implant removal are not well established. There are few definitive data to guide whether implant removal is appropriate. Implant removal may be challenging and lead to complications, such as neurovascular injury, refracture, or recurrence of deformity. When implants are removed for pain relief alone, the results are unpredictable and depend on both the implant type and its anatomic location. Current literature does not support the routine removal of implants to protect against allergy, carcinogenesis, or metal detection. Surgeons and patients should be aware of appropriate indications and have realistic expectations of the risks and benefits of implant removal. PMID- 16467187 TI - Carboxyl-terminal splicing enhances physical interactions between the cytoplasmic tails of purinergic P2X receptors. AB - Purinergic P2X receptors are ion-conducting channels composed of three subunits, each having two transmembrane domains and intracellular amino (N) and carboxyl (C) termini. Although alternative splicing extensively modifies the C-terminal sequences of P2X subunits, the direct influence of such post-transcriptional modifications on receptor architecture and function remains poorly understood. In this study, we focused on mouse pituitary P2X2 receptors. In this tissue, progressive splicing of the P2X2a C terminus generated two functional subunit variants, P2X2b and P2X2e, which exhibited accelerated desensitization rates and attenuated calcium signals when the receptors were in homomeric states. To measure the intersubunit interaction in living cells, the efficient transfer of bioluminescent resonance energy between luciferase and fluorescent proteins attached to the N- or C-subunit termini of these subunits was used. The constitutive interactions between the full-length C termini of P2X2a receptor were detected by a significant increase in fluorescence/luminescence intensity ratio compared with negative controls. Moreover, interactions between C termini and between C- and N termini of adjacent subunits were significantly enhanced in homomeric and heteromeric receptors containing P2X2b or P2X2e subunits. Finally, deletion of two amino acids at the splicing junction, but not at the C-terminal end of the P2X2b receptor, resulted in the enhancement of channel desensitization and luminescence resonance energy transfer. These results indicate that C terminal structure plays a critical role in the cytoplasmic intersubunit interactions and suggest that the extent of subunit interactions before ATP application could contribute to the subsequent channel activity and conformation changes associated with agonist-dependent desensitization. PMID- 16467188 TI - Dioxin induces an estrogen-like, estrogen receptor-dependent gene expression response in the murine uterus. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a ubiquitous environmental contaminant that elicits a broad range of toxicities in a tissue-, sex-, age-, and species-specific manner, including alterations in estrogen signaling. Many, if not all, of these effects involve changes in gene expression mediated via the activation of the aryl hydrocarbon receptor (AhR), a ligand activated transcription factor. Recent data indicate that TCDD may also elicit AhR-mediated estrogenic activity through interactions with the estrogen receptor (ER). In an effort to further characterize the estrogenic activity of TCDD, a comprehensive time-course analysis of uterine gene expression was conducted using ovariectomized C57BL/6 mice. Comparison of the temporal uterine transcriptional response to TCDD with that of ethynyl estradiol (EE) revealed a large proportion of the TCDD-mediated gene expression changes were also responsive to EE. Furthermore, pretreatment of mice with the pure ER antagonist ICI 182 780 (faslodex) inhibited gene expression responses to both EE and TCDD, providing additional evidence that these transcriptional responses involve the ER. PMID- 16467189 TI - Mechanisms underlying lipopolysaccharide-induced kinin B1 receptor up-regulation in the pig iris sphincter in vitro. AB - Kinin B1 receptors are known to be highly induced after inflammatory stimuli in several biological systems. We report that incubation of pig iris sphincter with lipopolysaccharide from Escherichia coli caused a marked and time-related up regulation of B1, accompanied by a reduction of B2 receptor-mediated contractile responses. The up-regulation of B1 receptors by lipopolysaccharide stimulation was decreased by the inhibitors of protein synthesis, cycloheximide and actinomycin D, and by dexamethasone and the nuclear factor-kappaB (NF-kappaB) inhibitor pyrrolidinedithiocarbamate (PDTC). In addition, lipopolysaccharide induced up-regulation of B1 receptors in the pig iris sphincter was significantly reduced by the p38 inhibitor 4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4 pyridyl)1H-imidazole (SB203580) and to a lesser extent by the extracellular signal-regulated kinases 1 and 2 (ERK1/2) blocker 2'-amino-3'-methoxyflavone (PD98059). Molecular biology experiments demonstrated that in vitro incubation with lipopolysaccharide resulted in a time-dependent and remarkable activation of NF-kappaB and of p38 and ERK1/2 mitogen-activated protein (MAP) kinases, in pig iris sphincter preparations. While attempting to verify how MAP kinases are part of the B1 receptor-activated signaling transduction pathways, we observed that PD98059 was able to markedly reduce the contraction induced by B1 receptor activation in lipopolysaccharide-pretreated pig iris sphincter muscle but that this response was only partially decreased by SB203580. Our results extend the previous evidence on the mechanisms underlying the B1 receptor upregulation processes and demonstrate for the first time how this takes place in an ocular tissue, the pig iris sphincter. It is therefore possible to define B1 receptors as therapeutic targets for the treatment of infectious and inflammatory alterations of the eye. PMID- 16467190 TI - Celecoxib inhibits interleukin-12 alphabeta and beta2 folding and secretion by a novel COX2-independent mechanism involving chaperones of the endoplasmic reticulum. AB - Celecoxib (CE) is a nonsteroidal anti-inflammatory drug (NSAID) that is a specific inhibitor of cyclooxygenase 2 (COX2). It is indicated for a variety of chronic inflammatory conditions, including rheumatoid arthritis. Over the last few years, adverse cardiovascular effects and increased risk for heart attacks have been associated with this drug. In addition, evidence is emerging for COX2 independent molecular targets. CE has been shown to induce apoptosis in various cancer cells lines through a COX2-independent mechanism that seems to involve inactivation of protein kinase Akt and inhibition of endoplasmic reticulum (ER) Ca2+ ATPase. In this study, we show that both CE and an analog devoid of COX2 inhibitory activity [1-(4-sulfamoyl phenyl)-3-trifluoromethyl-5-(4 trifluoromethylphenyl)pyrazole, CEA] inhibit the secretion of the dimeric interleukin-12 (IL-12) alphabeta and beta2 forms with identical IC50 values of 20 and 30 microM, respectively, whereas no such effect was seen with rofecoxib. Reverse transcription-polymerase chain reaction analysis showed that this inhibition was not due to a blockage of transcription of the alpha- and beta chain expression cassettes. Secretion of the beta monomer form was less strongly inhibited, suggestive for a mechanism primarily targeting dimer assembly in the ER. Analysis of intracellular fractions revealed that both CE and CEA increased the association of IL-12 with calreticulin, an endoplasmic reticulum-resident chaperone involved in the retention of misfolded cargo proteins while blocking interaction with ERp44. Our findings reveal a previously undescribed effect of celecoxib on oligomer protein folding and assembly in the endoplasmic reticulum and ensuing secretion and suggest that celecoxib-driven alteration of the secretome may be involved in some of its clinical side effects. PMID- 16467191 TI - Allosteric modulation of binding properties between units of chemokine receptor homo- and hetero-oligomers. AB - We have demonstrated previously that the chemokine receptors CCR2 and CCR5 form homo- and heterodimers and that dimers can only bind a single chemokine molecule with high affinity. We provide here evidence from bioluminescence resonance energy transfer experiments that stimulation by chemokines does not influence the CCR2/CCR5 heterodimerization status. In addition, we show that the rate of radioligand dissociation from one unit of the heterodimer in "infinite" tracer dilution conditions is strongly increased in the presence of an unlabeled chemokine ligand of the other unit. These results demonstrate unambiguously that the interaction between heterodimer units is of allosteric nature. Agonists, but also some monoclonal antibodies, could promote such negative binding cooperativity, indicating that this phenomenon does not require the full conformational change associated with receptor activation. Finally, we show that G protein coupling is required for high-affinity binding of macrophage inflammatory protein-1beta (CCL4) to CCR5 and that the dissociation from G proteins, after incubation with Gpp(NH)p, promotes the release of prebound radiolabeled chemokines with kinetics similar to those measured after the addition of an excess of unlabeled chemokines. These observations suggest that the association with G proteins probably participates in the negative cooperativity observed between receptor monomers. We propose that negative cooperativity within homo- and heterodimers of chemokine receptors and probably other G protein-coupled receptors will probably have major implications in their pharmacology in vivo and in the physiopathology of the diseases with which they are associated. PMID- 16467192 TI - Focus issue: signaling lipids. AB - Membranes are dynamic and specific contributors to cell signaling. Cellular membranes play a key structural role in creating sites for the formation of signaling complexes. Changes in membrane phospholipids can regulate the activity of transmembrane and peripheral membrane proteins. Modification of membrane lipids can result in formation of dynamic signaling molecules. Science's STKE highlights new insights into the roles that lipids and membranes play in cell signaling. PMID- 16467193 TI - Role of docosahexaenoic acid in neuronal plasma membranes. AB - The omega-3 fatty acid docosahexaenoic acid (DHA n-3) has long been known to be a major component of phosphoglycerides in the gray matter of mammalian brains. Furthermore, early studies of synaptosomes that had been isolated from gray matter showed that the plasma membranes of the synaptosomes contained DHA n-3 that was selectively esterified to phosphatidylethanolamine, plasmenylethanolamine (alkenylacyl-glycero-phosphorylethanolamine), and phosphatidylserine. In contrast, the phosphatidylcholine in these membranes contained esterified oleic acid, and the sphingomyelin and glycolipids in the membranes contained amide-linked stearic acid instead of a mixture of this acid with other, amide-linked fatty acids. The full implications of this unusual distribution of lipid head groups, esterified fatty acids, and amide-linked fatty acids are unclear, but the phosphoglycerides and sphingosine-containing lipids appear to be distributed asymmetrically between the two leaflets of the plasma membrane lipid bilayer and are likely to contribute to a dynamic lipid substructure. Because very few neuronal plasma membranes have been isolated and characterized to date, a major challenge for the future will be to investigate the composition of the lipid bilayers of different neuronal plasma membranes and identify effects of DHA n-3-containing phosphoglycerides on the ability of the plasma membranes to perform their many different functions. The aim of this Perspective is to stimulate further work in this important area by discussing recent evidence related to the role of neuronal plasma membrane phosphoglycerides in cell signaling. PMID- 16467194 TI - Building signaling complexes at the membrane. AB - Cell membranes, the plasma membrane in particular, serve as the hub of various cellular signaling networks. In response to receptor activation, many cytosolic proteins are reversibly recruited to membranes to form dynamic signaling complexes. Membranes provide a unique microenvironment in which the formation of signaling complexes can be exquisitely modulated by the fine interplay between protein-protein and lipid-protein interactions. PMID- 16467195 TI - Requirement of the human T-cell leukemia virus (HTLV-1) tax-stimulated HIAP-1 gene for the survival of transformed lymphocytes. AB - Human T cell leukemia virus type 1 (HTLV-1), the cause of adult T cell leukemia (ATL), induces clonal expansion of infected T-cells in nonleukemic individuals and immortalizes T cells in vitro. The resistance against apoptotic stimuli of these cells hints at a viral survival function in addition to a proliferation stimulating activity. Here we describe the up-regulation of the antiapoptotic HIAP-1/CIAP-2 gene as a consistent phenotype of HTLV-1-transformed and ATL derived cultures and its stimulation by the viral oncoprotein Tax. Cotransfections revealed a 60-fold increase of HIAP-1 promoter activity mediated by Tax mainly via nuclear factor-kappaB (NF-kappaB) activation. To address the relevance of virally increased HIAP-1 levels for the survival of HTLV-1 transformed cells, its expression was RNA interference (RNAi) suppressed using a lentiviral transduction system. This resulted in a dramatic reduction of cell growth, a strong induction of apoptosis rates, and increased caspases 3/7 activity, which is known to be suppressed by HIAP-1. Thus, the Tax-mediated HIAP 1 overexpression is required to suppress endogenous apoptosis and, therefore, is essential for the survival of HTLV-1-transformed lymphocytes. Moreover, this points to HIAP-1 as an important target of the HTLV-1-mediated NF-kappaB activation. PMID- 16467196 TI - SHIP deficiency enhances HSC proliferation and survival but compromises homing and repopulation. AB - The SH2 domain-containing inositol 5'-phosphatase-1 (SHIP) has the potential to modulate multiple signaling pathways downstream of receptors that impact hematopoietic stem cell (HSC) biology. Therefore, we postulated that SHIP might play an important role in HSC homeostasis and function. Consistent with this hypothesis, HSC proliferation and numbers are increased in SHIP(-/-) mice. Despite expansion of the compartment, SHIP(-/-) HSCs exhibit reduced capacity for long-term repopulation. Interestingly, we observe that SHIP(-/-) stem/progenitor cells home inefficiently to bone marrow (BM), and consistent with this finding, have reduced surface levels of both CXCR4 and vascular cell adhesion marker-1 (VCAM-1). These studies demonstrate that SHIP is critical for normal HSC function, homeostasis, and homing. PMID- 16467197 TI - Upfront double high-dose chemotherapy with DICEP followed by BEAM and autologous stem cell transplantation for poor-prognosis aggressive non-Hodgkin lymphoma. AB - A single center, prospective clinical trial was conducted evaluating 2 cycles of induction high-dose chemotherapy for adults younger than 65 years of age with aggressive non-Hodgkin lymphoma (NHL) and 2 to 3 Age-Adjusted International Prognostic Index risk factors. Patients received one cycle of standard dose cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) followed by one cycle of dose-intensive cyclophosphamide 5.25 g/m(2), etoposide 1.05 g/m(2), cisplatin 105 mg/m(2) (DICEP), then underwent autologous blood stem cell collection, followed by one cycle of high-dose carmustine (BCNU) 300 mg/m(2), etoposide 800 mg/m(2), Ara-C 1600 mg/m(2), melphalan 140 mg/m(2) (BEAM), and autologous stem cell transplantation (ASCT) and radiotherapy to prior bulk. From June 1998 to August 2004, 55 patients aged 20 to 63 years (median 44 years) were accrued, 51 (92%) of whom had diffuse large B-cell NHL. Poor prognostic factors included stage 4 (n = 46), elevated lactate dehydrogenase (LDH; n = 47), Eastern Cooperative Oncology Group (ECOG) performance status 2 to 4 (n = 43), bulky mass more than 10 cm (n = 34), and marrow involvement (n = 16). Only one patient experienced nonrelapse mortality. With a median follow-up of 49 months, 4-year event-free survival (EFS) and overall survival (OS) rates for all 55 patients are 72% (95% confidence interval [CI] = 60%-84%) and 79% (95% CI = 69%-90%), respectively. In conclusion, CHOP-DICEP-BEAM is feasible and gave encouraging EFS and OS for patients with poor-prognosis aggressive NHL. PMID- 16467198 TI - HIV nonprogressors preferentially maintain highly functional HIV-specific CD8+ T cells. AB - Establishing a CD8(+) T cell-mediated immune correlate of protection in HIV disease is crucial to the development of vaccines designed to generate cell mediated immunity. Historically, neither the quantity nor breadth of the HIV specific CD8(+) T-cell response has correlated conclusively with protection. Here, we assess the quality of the HIV-specific CD8(+) T-cell response by measuring 5 CD8(+) T-cell functions (degranulation, IFN-gamma, MIP-1beta, TNF alpha, and IL-2) simultaneously in chronically HIV-infected individuals and elite nonprogressors. We find that the functional profile of HIV-specific CD8(+) T cells in progressors is limited compared to that of nonprogressors, who consistently maintain highly functional CD8(+) T cells. This limited functionality is independent of HLA type and T-cell memory phenotype, is HIV specific rather than generalized, and is not effectively restored by therapeutic intervention. Whereas the total HIV-specific CD8(+) T-cell frequency did not correlate with viral load, the frequency and proportion of the HIV-specific T cell response with highest functionality inversely correlated with viral load in the progressors. Thus, rather than quantity or phenotype, the quality of the CD8(+) T-cell functional response serves as an immune correlate of HIV disease progression and a potential qualifying factor for evaluation of HIV vaccine efficacy. PMID- 16467199 TI - BCR-ABL mRNA levels at and after the time of a complete cytogenetic response (CCR) predict the duration of CCR in imatinib mesylate-treated patients with CML. AB - Although most patients with chronic myeloid leukemia (CML) treated with imatinib mesylate achieve a complete cytogenetic response (CCR), some patients will relapse. To determine the potential of real-time quantitative BCR-ABL reverse transcriptase-polymerase chain reaction (RT-PCR) to predict the duration of continued CCR, we monitored 85 patients treated with imatinib mesylate who achieved a CCR. With a median follow-up of 13 months after CCR (29 months after imatinib mesylate; median 6 RQ-PCR assays), 23 patients (27%) had disease progression (predominantly loss of CCR). Compared with the median baseline level of BCR-ABL mRNA, 42% of patients achieved at least a 2-log molecular response at the time of first reaching CCR. Failure to achieve a 2-log response at the time of CCR was an independent predictive marker of subsequent progression-free survival (hazard ratio = 5.8; 95% CI, 1.7-20; P = .005). After CCR, BCR-ABL mRNA levels progressively declined for at least the next 15 months, and 42 patients (49%) ultimately achieved at least a 3-log reduction in BCR-ABL mRNA. Patients failing to achieve this 3-log response, at any time during therapy, had significantly shorter progression-free survival (hazard ratio = 8.1; 95% CI, 3.1 22; P < .001). The achievement of either a 2-log molecular response at the time of CCR or a 3-log response anytime thereafter is a significant and independent prognostic marker of subsequent progression-free survival. PMID- 16467200 TI - The inhibitory anti-FGFR3 antibody, PRO-001, is cytotoxic to t(4;14) multiple myeloma cells. AB - The association of fibroblast growth factor receptor 3 (FGFR3) expression with t(4;14) multiple myeloma (MM) and the demonstration of the transforming potential of this receptor tyrosine kinase (RTK) make it a particularly attractive target for drug development. We report here a novel and highly specific anti-FGFR3 neutralizing antibody (PRO-001). PRO-001 binds to FGFR3 expressed on transformed cells and inhibits FGFR3 autophosphorylation and downstream signaling. The antibody inhibited the growth of FGFR3-expressing FDCP cells (IC(50) of 0.5 microg/mL) but not that of cells expressing FGFR1 or FGFR2, and potently inhibited FGFR3-dependent solid tumor growth in a mouse xenograft model. Furthermore, PRO-001 inhibited the growth of the FGFR3-expressing, human myeloma cell line, UTMC2. Inhibition of viability was still observed when cells were cocultured with stroma or in the presence of IL-6 or IGF-1. PRO-001 did not inhibit constitutive activation of K650E, G384D, and Y373C FGFR3 in myeloma cell lines and failed to inhibit the growth of these cells. Most importantly, however, PRO-001 induced cytotoxic responses in primary t(4;14)(+) MM samples with an increase in apoptotic index of 20% to 80% as determined by annexin V staining. The data demonstrate that PRO-001 is a potent and specific inhibitor of FGFR3 and deserves further study for the treatment of FGFR3-expressing myeloma. PMID- 16467201 TI - Cytokine-induced IL-10-secreting CD8 T cells represent a phenotypically distinct suppressor T-cell lineage. AB - Populations of regulatory T cells (Tregs) control autoimmune and allergic immunopathology induced by self or foreign antigens. Several types of CD4(+) MHC class II-restricted Treg populations have been characterized, but the biology of CD8(+), MHC class I-restricted Tregs is less understood. We show here that CD8(+) Tregs are rapidly generated in the presence of IL-4 and IL-12, produce IL-10, and exhibit a unique cell-surface phenotype with coexpression of activation and naive cell-associated markers. They block activation of naive or effector T cells and suppress IgG/IgE antibody responses and graft-versus-host disease in vivo. Suppression is dependent on cell contact and mediated by direct T-cell-T-cell interaction that antagonizes T-cell-receptor (TCR) signals. The data establish the existence of a CD8 T-cell suppressor effector subset distinct in both phenotype and function from T cytotoxic 1 (Tc1) and Tc2 cells. Production of such CD8 Tregs has potential for cell-based therapy of CD4 or CD8 T-cell-mediated disease. PMID- 16467202 TI - Deficiency in CCR5 but not CCR1 protects against neointima formation in atherosclerosis-prone mice: involvement of IL-10. AB - The chemokine RANTES has been implicated in neointimal hyperplasia after arterial injury. We analyzed the differential role of the RANTES receptors CCR1 and CCR5 by genetic deletion in apolipoprotein E-deficient mice. Deficiency in CCR5 significantly reduced neointimal area after arterial wire injury, associated with a decrease in macrophages, CD3(+) T lymphocytes, and CCR2(+) cells. In contrast, CCR1 deficiency did not affect neointimal area or cell content. Deletion of CCR5 entailed an up-regulation of the anti-inflammatory cytokine interleukin 10 (IL 10) in neointimal smooth muscle cells, and its antibody blockade reversed effects in CCR5(-/-) mice. Conversely, proinflammatory interferon gamma was increased in the neointima of CCR1(-/-) mice, and its blockade unmasked a reduction in macrophage recruitment. Our data indicate that CCR5 is more crucial than CCR1 for neointimal plaque formation, and that its attenuation in CCR5(-/-) mice is due to an atheroprotective immune response involving IL-10. This harbors important implications for targeting chemokine receptors in vascular remodeling. PMID- 16467203 TI - Platelet depletion in mice increases mortality after thermal injury. AB - Platelets play a fundamental role in maintaining hemostasis and have been shown to participate in innate and adaptive immunity. However, the role of platelets in the immune response to injury remains undefined. We tested the importance of platelets in the host response to serious injury in a newly developed platelet deficient mouse model. Wild-type and platelet-depleted C57BL/6J mice underwent a 25% full-thickness total body surface area thermal or sham injury. Platelet deficient mice showed survival of 51% at 48 hours after injury compared with 94% to 100% survival in experimental control mice (P < .001). Necropsy and histology ruled out hemorrhage and hypovolemia as causes of death. Percentages of peripheral blood monocytes (P < .01) and neutrophils (P < .05) were increased between 36 and 48 hours after thermal injury in platelet-deficient mice compared with control mice. Plasma levels of TNFalpha (P < .001), IL-6 (P < .001), and MCP 1 (P < .05) were also elevated by 24 hours whereas levels of TGFbeta(1) were reduced between 24 and 36 hours following injury in platelet-depleted mice (P < .001) compared with control mice. Our findings demonstrate for the first time that platelets play a critical protective role during the host response to injury. Moreover, our findings suggest that platelets and, more importantly, platelet-derived TGFbeta(1) modulate the systemic inflammatory response occurring after injury. PMID- 16467204 TI - Cul4A targets p27 for degradation and regulates proliferation, cell cycle exit, and differentiation during erythropoiesis. AB - As erythroid progenitors differentiate into precursors and finally mature red blood cells, lineage-specific genes are induced, and proliferation declines until cell cycle exit. Cul4A encodes a core subunit of a ubiquitin ligase that targets proteins for ubiquitin-mediated degradation, and Cul4A-haploinsufficient mice display hematopoietic dysregulation with fewer multipotential and erythroid committed progenitors. In this study, stress induced by 5-fluorouracil or phenylhydrazine revealed a delay in the recovery of erythroid progenitors, early precursors, and normal hematocrits in Cul4A(+/-) mice. Conversely, overexpression of Cul4A in a growth factor-dependent, proerythroblast cell line increased proliferation and the proportion of cells in S phase. When these proerythroblasts were induced to terminally differentiate, endogenous Cul4A protein expression declined 3.6-fold. Its enforced expression interfered with erythrocyte maturation and cell cycle exit and, instead, promoted proliferation. Furthermore, p27 normally accumulates during erythroid terminal differentiation, but Cul4A enforced expression destabilized p27 and attenuated its accumulation. Cul4A and p27 proteins coimmunoprecipitate, indicating that a Cul4A ubiquitin ligase targets p27 for degradation. These findings indicate that a Cul4A ubiquitin ligase positively regulates proliferation by targeting p27 for degradation and that Cul4A down-regulation during terminal erythroid differentiation allows p27 to accumulate and signal cell cycle exit. PMID- 16467205 TI - Integrin inhibition through Lyn-dependent cross talk from CXCR4 chemokine receptors in normal human CD34+ marrow cells. AB - We studied the effects of Lyn ablation on CXCR4 receptor-mediated migration and adhesion of hematopoietic precursors. Down-regulation of Lyn expression with siRNA greatly reduced CXCR4-dependent hematopoietic cell movement, and increased cell adherence to stroma. This increase was associated with the up-regulated expression of activation-dependent epitopes of the beta(2) integrin LFA-1 and was prevented by antibodies that selectively block cell adhesion mediated by ICAM-1. Attachment to surfaces coated with ICAM-1 was also enhanced in Lyn-depleted hematopoietic cells, as compared with Lyn-expressing cells. Functional rescue experiments with Lyn siRNA targeting the 3' UTR indicated that the observed effects can be attributed directly to specific inhibition of Lyn. Our results show that in chemokine-stimulated hematopoietic cells Lyn kinase is a positive regulator of cell movement while negatively regulating adhesion to stromal cells by inhibiting the ICAM-1-binding activity of beta(2) integrins. These results provide a molecular mechanism for cross talk between the chemokine receptor CXCR4 and beta(2) integrins. This cross talk may allow chemokine receptors to modulate the arrest of rolling hematopoietic precursors on the surface of bone marrow stromal cells. PMID- 16467206 TI - Cytochrome c-related caspase-3 activation determines treatment response and relapse in childhood precursor B-cell ALL. AB - Deficient activation of apoptosis signaling pathways may be responsible for treatment failure in acute leukemia. Here, we address the impact of intact apoptosis signaling in 78 patients with pediatric precursor B-cell acute lymphoblastic leukemia (ALL) by analysis of 2 key apoptogenic events: caspase-3 activation and cytochrome c release in leukemia cells cultured in vitro. Both events correlated only in the group of patients who had a good response and patients in continuous remission, suggesting that intact apoptosis signaling is a characteristic for favorable outcome. By combining both parameters, we identified a novel indicator, cytochrome c-related activation of caspase-3 (CRAC). CRAC directly connects the extent of caspase-3 activation to cytochrome c release in single cells in an individual patient sample. In CRAC-positive patients, indicating proficient apoptosis signaling, the number of persisting leukemia cells on day 15 was significantly lower than in the CRAC-negative patient group (n = 27, mean 6.0% versus n = 36, mean 22.6%; P = .003). At a median follow-up of 31 months, disease-free survival was 84 months (95% CI = 76 to 91 months) and 66 months (95% CI = 52 to 80 months) for patients with positive and negative CRAC, respectively (P = .019). CRAC may serve as a functionally defined risk factor for treatment stratification. PMID- 16467207 TI - Wilms tumor suppressor, Wt1, is a transcriptional activator of the erythropoietin gene. AB - Molecular mechanisms for the developmental stage and tissue-specific regulation of the erythropoietin (EPO) gene are poorly understood. Recent findings indicate a role of the Wilms tumor suppressor, Wt1, in the formation of the hematopoietic system. Herein, we tested the hypothesis that Wt1 is a transcriptional regulator of the EPO gene. Binding of the transcriptionally competent Wt1(-KTS) isoform to the minimal EPO promoter was demonstrated by electrophoretic mobility shift assay and chromatin immunoprecipitation. Under normoxia, EPO expression was significantly increased in HEK 293 and HepG2 cells with forced expression of Wt1( KTS). A reporter construct harboring the 117-bp minimal human EPO promoter was activated up to 20-fold by transient cotransfection of Wt1(-KTS) in different cell lines. Mutation of the Wt1 binding site in the EPO promoter abrogated this stimulatory effect of the Wt1(-KTS) protein. Hepatic Epo mRNA expression was significantly reduced in embryonic mice with homozygous Wt1 deletion. Furthermore, Wt1 and EPO were colocalized in hepatocytes of the liver and in neuronal cells of the dorsal root ganglia in developing mice. Both proteins were also detected in Sertoli cells of the adult murine testis. In conclusion, we identified Wt1(-KTS) as a novel transcriptional activator for the tissue-specific expression of the EPO gene. PMID- 16467208 TI - MEK1 inhibition sensitizes primary acute myelogenous leukemia to arsenic trioxide induced apoptosis. AB - We found that MEK1 inhibitor PD184352 strikingly increased apoptosis induced by arsenic trioxide (ATO) in 21 of 25 patients with primary acute myelogenous leukemia (AML). Isobologram analysis confirmed the synergistic (13 of 25 patients) or additive (8 of 25 patients) nature of this interaction. Moreover, we demonstrated that the p53-related gene p73 is a molecular target of the combined treatment in AML blasts. Indeed, ATO modulates the expression of the p73 gene by inducing the proapoptotic and antiproliferative TAp73 and the antiapoptotic and proproliferative DeltaNp73 isoforms, thereby failing to elevate the TA/DeltaNp73 ratio. Conversely, treatment with PD184352 reduces the level of DeltaNp73 and blunts the arsenic-mediated up-regulation of DeltaNp73, thus causing an increase in the TA/DeltaNp73 ratio of dual-treated cells. High doses of ATO induced p53 accumulation in 11 of 21 patients. Combined treatment resulted in the induction of the proapoptotic p53/p73 target gene p53AIP1 (p53-regulated apoptosis-inducing protein 1) and greatly enhanced the apoptosis of treated cells. PMID- 16467209 TI - Comparison of 3.0- and 1.5-T three-dimensional time-of-flight MR angiography in moyamoya disease: preliminary experience. AB - PURPOSE: To prospectively compare 3.0- and 1.5-T three-dimensional (3D) time-of flight (TOF) magnetic resonance (MR) angiography in patients with moyamoya disease, with special emphasis on the visualization of abnormal netlike vessels (moyamoya vessels). MATERIALS AND METHODS: Study protocols were approved by the local ethics committee; written informed consent was obtained from all patients. The study included 24 consecutive patients with moyamoya disease (four male and 20 female patients). Patients ranged in age from 17 to 66 years (mean age, 41 years). Moyamoya disease had been diagnosed in all patients before they were entered into the study. All patients underwent 3D TOF MR angiography at both 3.0 and 1.5 T; imaging examinations were performed within 14 days of each other. Maximum intensity projections (MIPs) obtained with MR angiography performed at both 3.0 and 1.5 T were evaluated by two neuroradiologists; the visualization of moyamoya vessels was graded according to a 4-point scale. For both 3.0- and 1.5-T imaging, the number of high-signal-intensity areas and the summation of cross sectional areas of high signal intensity on source images obtained at the same level of MR angiography were compared quantitatively by using the Wilcoxon matched-pair signed-rank test. RESULTS: Moyamoya vessels were better visualized on MIPs obtained with 3.0-T imaging than on MIPs obtained with 1.5-T imaging (P < .001). At the identical level of the source image, 3.0-T imaging depicted more high-signal-intensity areas than did 1.5-T imaging. Wider cross-sectional areas of moyamoya vessels were visualized with 3.0-T imaging than with 1.5-T imaging (P < .001). CONCLUSION: Moyamoya vessels are better depicted with MR angiography at 3.0 T than at 1.5 T. PMID- 16467210 TI - Pulmonary nodules: estimation of malignancy at thin-section helical CT--effect of computer-aided diagnosis on performance of radiologists. AB - PURPOSE: To evaluate the effect of a computer-aided diagnosis (CAD) system on the diagnostic performance of radiologists for the estimation of the malignancy of pulmonary nodules on thin-section helical computed tomographic (CT) scans. MATERIALS AND METHODS: The institutional review board approved use of the CT database; informed specific study-related consent was waived. The institutional review board approved participation of radiologists; informed consent was obtained from all observers. Thirty-three (18 malignant, 15 benign) pulmonary nodules of less than 3.0 cm in maximal diameter were evaluated. Receiver operating characteristic (ROC) analysis with a continuous rating scale was used to compare observer performance for the estimation of the likelihood of malignancy first without and then with the CAD system. The participants were 10 board-certified radiologists and nine radiology residents. RESULTS: For all 19 participants, the mean area under the best-fit ROC curve (A(z)) values achieved without and with the CAD system were 0.843 +/- 0.097 (standard deviation) and 0.924 +/- 0.043, respectively. The difference was significant (P = .021). The mean A(z) values achieved without and with the CAD system were 0.910 +/- 0.052 and 0.944 +/- 0.040, respectively, for the 10 board-certified radiologists (P = .190) and 0.768 +/- 0.078 and 0.901 +/- 0.036, respectively, for the nine radiology residents (P = .009). CONCLUSION: Use of the CAD system significantly (P = .009) improved the diagnostic performance of radiology residents for assessment of the malignancy of pulmonary nodules; however, it did not improve that of board-certified radiologists. PMID- 16467211 TI - Preoperative assessment of resectability of hepatic hilar cholangiocarcinoma: combined CT and cholangiography with revised criteria. AB - PURPOSE: To retrospectively assess the accuracy of combined multiphasic computed tomography (CT) and direct cholangiography for evaluation of the resectability of hilar cholangiocarcinoma, on the basis of revised criteria for unresectability, by using surgery as the reference standard. MATERIALS AND METHODS: Institutional review board approval was obtained, and informed consent was waived. From 1998 to 2003, 55 patients (37 men, 18 women; mean age +/- standard deviation, 59 years +/ 12) with surgically proved hilar cholangiocarcinomas who underwent preoperative CT (single-detector row CT, n = 26; multi-detector row CT, n = 29) and cholangiography were included for study. The authors' revised criteria for unresectable tumor were contralateral hepatic artery invasion; main or contralateral portal vein invasion longer than 2 cm; biliary extension to the contralateral secondary confluence, farther than 2 cm from hepatic hilum; enlarged lymph nodes at the celiac, portacaval, and paraaortic area; and other ancillary findings. Tumor resectability based on these parameters was determined at imaging by two radiologists in consensus. Mann-Whitney U test and weighted kappa coefficient of agreement were used for accuracy determination. RESULTS: For depiction of portal vein invasion (in 26 patients), CT yielded an accuracy of 85.5%. Arterial invasion was found at surgery in 19 patients, with CT providing an accuracy of 92.7%. For prediction of node involvement (15 patients, 27%), CT yielded an accuracy of 83.6%. The extent of ductal involvement could be accurately predicted in 46 patients (84%) (weighted kappa = 0.767). In 30 of 42 patients with disease classified as resectable according to revised criteria, disease was found to be resectable at surgery (71.4% positive predictive value). In 11 of 13 patients with disease classified as unresectable according to revised criteria, unresectable disease was confirmed (84.6% negative predictive value). Overall accuracy of resectability was 74.5%. CONCLUSION: Combined interpretation of CT and direct cholangiographic images by using our revised criteria resulted in overall accuracy of 74.5% for prediction of resectability for hilar cholangiocarcinoma. PMID- 16467212 TI - Scrotal US for evaluation of infertile men with azoospermia. AB - PURPOSE: To evaluate prospectively the accuracy of scrotal ultrasonography (US) in distinguishing obstructive azoospermia from nonobstructive azoospermia in infertile men by using histologic findings as the reference standard. MATERIALS AND METHODS: The institutional review board approved the study, and informed consent was obtained from each patient. Twenty infertile men (mean age, 34.7 years; 40 testes) with azoospermia were evaluated at scrotal US, with an emphasis on the course of the proximal genital duct: mediastinum testis, epididymal head, epididymal body, and epididymal tail. Testicular volumes were calculated by using the formula: length x height x width x 0.71. On the basis of histologic results, azoospermia was divided into two groups (obstructive vs nonobstructive) in all cases except one. Scrotal US findings between obstructive and nonobstructive azoospermia were compared. The Fisher exact and Wilcoxon signed rank sum tests were used to assess differences between both groups. RESULTS: Of 20 infertile men with azoospermia, 14 were proved to have obstructive azoospermia; the others had nonobstructive azoospermia. According to US findings, epididymal abnormalities in the head, body, and tail were significantly associated with obstructive azoospermia (17 [61%], 18 [64%], and 20 [71%] of 28 testes, respectively; P < .001 for all), while abnormalities of the mediastinum testis between both groups were not significant (P > .05). By taking epididymal abnormalities into account, sensitivity, specificity, and accuracy of scrotal US for differentiation of obstructive from nonobstructive azoospermia were 82.1% (23 of 28 scrota), 100% (12 of 12 scrota), and 87.5% (35 of 40 scrota), respectively. The median testicular volume in obstructive azoospermia was 11.6 mL (range, 7.7-25.8 mL) and that in nonobstructive azoospermia was 8.3 mL (range, 1.2-16.4 mL) (P < .05). CONCLUSION: Evaluation of the epididymis and measurement of testicular volume with scrotal US are important in distinguishing obstructive azoospermia from nonobstructive azoospermia in infertile men. PMID- 16467213 TI - In response to Cadet et al. PMID- 16467214 TI - Association of TPH1 with suicidal behaviour and psychiatric disorders in the Chinese population. AB - Tryptophan hydroxylase (TPH), the rate limiting enzyme in serotonin biosynthesis, is one of the most important regulating factors in the serotonergic system. Recently, polymorphisms of the TPH gene have been identified as being associated with suicide, but the evidence is inconsistent. To investigate the role in suicide of one of the isoforms, TPH1, we examined the association of five single nucleotide polymorphisms (SNPs) in the promoter region and in intron 7 of the TPH1 gene based on a sample from the Chinese population of 810 subjects, of whom 329 had made no suicide attempts (NSA), 297 had made suicide attempts (SA), and 184 were healthy subjects (HS). In this study, we observed statistically significant differences between NSA and HS subjects in allele distributions on one marker, -6526A (p = 0.0329; odds ratio (OR) 1.36; 95% confidence interval (CI) 1.01 to 1.81). No significant difference in genotype distribution or allele frequencies of other polymorphisms was found between the suicide victims and the controls. The overall haplotype frequency was significantly different between cases and healthy controls (p = 0.000024 NSA v HS; p < 0.000001, SA v HS; p < 0.000001, cases v HS). We found the haplotype TCAAA of -7180/-7065/-6526/218/779 to be strongly associated with suicidal behaviour and psychiatric disorders (p = 0.00243; OR = 1.62; 95% CI 1.17 to 2.24 and p = 0.018; OR = 1.41; 95% CI 1.05 to 1.91), which suggests an association of TPH1 with suicidal behaviour and indicates that TPH1 may play a significant role in the aetiology of psychiatric disorders in the Han Chinese population. PMID- 16467215 TI - Loss of desmoplakin isoform I causes early onset cardiomyopathy and heart failure in a Naxos-like syndrome. AB - BACKGROUND: Desmosomes are cellular junctions important for intercellular adhesion and anchoring the intermediate filament (IF) cytoskeleton to the cell membrane. Desmoplakin (DSP) is the most abundant desmosomal protein with 2 isoforms produced by alternative splicing. METHODS: We describe a patient with a recessively inherited arrhythmogenic dilated cardiomyopathy with left and right ventricular involvement, epidermolytic palmoplantar keratoderma, and woolly hair. The patient showed a severe heart phenotype with an early onset and rapid progression to heart failure at 4 years of age. RESULTS: A homozygous nonsense mutation, R1267X, was found in exon 23 of the desmoplakin gene, which results in an isoform specific truncation of the larger DSPI isoform. The loss of most of the DSPI specific rod domain and C-terminal area was confirmed by Western blotting and immunofluorescence. We further showed that the truncated DSPI transcript is unstable, leading to a loss of DSPI. DSPI is reported to be an obligate constituent of desmosomes and the only isoform present in cardiac tissue. To address this, we reviewed the expression of DSP isoforms in the heart. Our data suggest that DSPI is the major cardiac isoform but we also show that specific compartments of the heart have detectable DSPII expression. CONCLUSIONS: This is the first description of a phenotype caused by a mutation affecting only one DSP isoform. Our findings emphasise the importance of desmoplakin and desmosomes in epidermal and cardiac function and additionally highlight the possibility that the different isoforms of desmoplakin may have distinct functional properties within the desmosome. PMID- 16467216 TI - Independent replication and initial fine mapping of 3p21-24 in Asperger syndrome. AB - BACKGROUND: Asperger syndrome is characterised by abnormalities in social interaction as well as repetitive and stereotyped behaviours and interests. The trait is thought to display complex inheritance, but in a subset of families the inheritance resembles the autosomal dominant model. Linkage to 3p14-24 has recently been reported in Asperger syndrome in Finnish families with a maximum multipoint NPL(all) of 3.32 at D3S2432. METHODS: We have replicated linkage findings to 3p21-24 in 12 new extended Asperger syndrome families. Linkage analyses were performed separately for the 12 new families, and linkage and association analyses were also performed jointly with data from the original genome-wide screen. RESULTS: Best two point and multipoint logarithm of the odds (LOD) scores in analyses of both data sets were obtained at D3S2432 (NPL(all) = 3.83) with both subsets of families contributing to linkage. Association analysis of the combined data set produced a trend towards association with D3S2432 and D3S1619. CONCLUSIONS: This study further validates 3q21-24 as a candidate region for Asperger syndrome. PMID- 16467217 TI - Linkage analysis in a large Swedish family supports the presence of a susceptibility locus for adenoma and colorectal cancer on chromosome 9q22.32 31.1. AB - BACKGROUND: The best known hereditary colorectal cancer syndromes, familial adenomatous polyposis (FAP) and hereditary non-polyposis colorectal cancer (HNPCC), constitute about 2% of all colorectal cancers, and there are at least as many non-FAP, non-HNPCC cases where the family history suggests a dominantly inherited colorectal cancer risk. Recently, a locus on chromosome 9q22.2-31.2 was identified by linkage analysis in sib pairs with colorectal cancer or adenoma. METHODS: Linkage analysis for the suggested locus on chromosome 9 was carried out in an extended Swedish family. This family had previously been investigated but following the identification of adenomas in several previously unaffected family members, these subjects were now considered to be gene carriers. RESULTS: In the present study, we found linkage of adenoma and colorectal cancer to chromosome 9q22.32-31.1 with a multipoint LOD score of 2.4. We were also able to define the region for this locus to 7.9 cM between the markers D9S280 and D9S277. CONCLUSIONS: Our result supports the presence of a susceptibility locus predisposing to adenoma and colorectal cancer in this chromosomal region. PMID- 16467219 TI - Testing association between LRRK2 and Parkinson's disease and investigating linkage disequilibrium. AB - BACKGROUND: We and others recently identified the gene underlying PARK8 linked Parkinson's disease (PD). This gene, LRRK2, contains mutations that cause an autosomal dominant PD, including a mutation, G2019S, which is the most common PD causing mutation identified to date. Common genetic variability in genes that contain PD causing mutations has previously been implicated as a risk factor for typical sporadic disease. METHODS: We undertook a case-control association analysis of LRRK2 in two independent European PD cohorts using 31 tagging single nucleotide polymorphisms (tSNPs) and five potentially functional SNPs. To assess the structure of this locus in different populations, we have performed linkage disequilibrium (LD) analysis using these variants in a human diversity panel. RESULTS: We show that common genetic variability in LRRK2 is not associated with risk for PD in the European populations studied here. We also show inter population variability in the strength of LD across this locus. CONCLUSIONS: To our knowledge this is the first comprehensive analysis of common variability within LRRK2 as a risk factor for PD. PMID- 16467220 TI - The cover. Portrait of Thomas Bulwer. PMID- 16467218 TI - Connective tissue dysplasia in five new patients with NF1 microdeletions: further expansion of phenotype and review of the literature. AB - Approximately 5% of patients with neurofibromatosis type 1 (NF1) have deletions of the entire NF1 gene. The phenotype usually includes early onset, large number of neurofibromas, presence of congenital anomalies, cognitive deficiency, and variable dysmorphic features and growth abnormalities. Connective tissue abnormalities are not generally recognised as a part of NF1 microdeletion syndrome, but mitral valve prolapse, joint laxity, and/or soft skin on the palms have been reported in a few patients. We describe clinical findings in six newly diagnosed patients with NF1 microdeletions, five of whom presented with connective tissue abnormalities. A literature review of the clinical findings associated with NF1 microdeletion was also performed. Our report confirms that connective tissue dysplasia is common in patients with NF1 microdeletions. Given the potential for associated cardiac manifestation, screening by echocardiogram may be warranted. Despite the large number (>150) of patients with known NF1 microdeletions, the clinical phenotype remains incompletely defined. Additional reports of patients with NF1 microdeletions, including comprehensive clinical and molecular information, are needed to elucidate possible genotype-phenotype correlation. PMID- 16467221 TI - Researchers address use of performance-enhancing drugs in nonelite athletes. PMID- 16467222 TI - Do lackluster trial findings mean new avenues are needed for heart research? PMID- 16467223 TI - Shift seen in patterns of drug use among teens. PMID- 16467224 TI - Group launches peripheral arterial disease guidelines. PMID- 16467225 TI - FDA speeds smoking cessation drug review. PMID- 16467226 TI - Physician-scientist training. PMID- 16467227 TI - Critically ill patients and acute renal failure. PMID- 16467228 TI - Critically ill patients and acute renal failure. PMID- 16467229 TI - Cerebral palsy litigation. PMID- 16467230 TI - Paraganglioma syndrome and mutations of the SDHC gene. PMID- 16467231 TI - Early triple therapy vs mono or dual therapy for children with perinatal HIV infection. PMID- 16467232 TI - Low-fat dietary pattern and risk of invasive breast cancer: the Women's Health Initiative Randomized Controlled Dietary Modification Trial. AB - CONTEXT: The hypothesis that a low-fat dietary pattern can reduce breast cancer risk has existed for decades but has never been tested in a controlled intervention trial. OBJECTIVE: To assess the effects of undertaking a low-fat dietary pattern on breast cancer incidence. DESIGN AND SETTING: A randomized, controlled, primary prevention trial conducted at 40 US clinical centers from 1993 to 2005. PARTICIPANTS: A total of 48,835 postmenopausal women, aged 50 to 79 years, without prior breast cancer, including 18.6% of minority race/ethnicity, were enrolled. INTERVENTIONS: Women were randomly assigned to the dietary modification intervention group (40% [n = 19,541]) or the comparison group (60% [n = 29,294]). The intervention was designed to promote dietary change with the goals of reducing intake of total fat to 20% of energy and increasing consumption of vegetables and fruit to at least 5 servings daily and grains to at least 6 servings daily. Comparison group participants were not asked to make dietary changes. MAIN OUTCOME MEASURE: Invasive breast cancer incidence. RESULTS: Dietary fat intake was significantly lower in the dietary modification intervention group compared with the comparison group. The difference between groups in change from baseline for percentage of energy from fat varied from 10.7% at year 1 to 8.1% at year 6. Vegetable and fruit consumption was higher in the intervention group by at least 1 serving per day and a smaller, more transient difference was found for grain consumption. The number of women who developed invasive breast cancer (annualized incidence rate) over the 8.1-year average follow-up period was 655 (0.42%) in the intervention group and 1072 (0.45%) in the comparison group (hazard ratio, 0.91; 95% confidence interval, 0.83-1.01 for the comparison between the 2 groups). Secondary analyses suggest a lower hazard ratio among adherent women, provide greater evidence of risk reduction among women having a high-fat diet at baseline, and suggest a dietary effect that varies by hormone receptor characteristics of the tumor. CONCLUSIONS: Among postmenopausal women, a low-fat dietary pattern did not result in a statistically significant reduction in invasive breast cancer risk over an 8.1-year average follow-up period. However, the nonsignificant trends observed suggesting reduced risk associated with a low-fat dietary pattern indicate that longer, planned, nonintervention follow-up may yield a more definitive comparison. CLINICAL TRIALS REGISTRATION: ClinicalTrials.gov Identifier: NCT00000611. PMID- 16467233 TI - Low-fat dietary pattern and risk of colorectal cancer: the Women's Health Initiative Randomized Controlled Dietary Modification Trial. AB - CONTEXT: Observational studies and polyp recurrence trials are not conclusive regarding the effects of a low-fat dietary pattern on risk of colorectal cancer, necessitating a primary prevention trial. OBJECTIVE: To evaluate the effects of a low-fat eating pattern on risk of colorectal cancer in postmenopausal women. DESIGN, SETTING, AND PARTICIPANTS: The Women's Health Initiative Dietary Modification Trial, a randomized controlled trial conducted in 48,835 postmenopausal women aged 50 to 79 years recruited between 1993 and 1998 from 40 clinical centers throughout the United States. INTERVENTIONS: Participants were randomly assigned to the dietary modification intervention (n = 19,541; 40%) or the comparison group (n = 29,294; 60%). The intensive behavioral modification program aimed to motivate and support reductions in dietary fat, to increase consumption of vegetables and fruits, and to increase grain servings by using group sessions, self-monitoring techniques, and other tailored and targeted strategies. Women in the comparison group continued their usual eating pattern. MAIN OUTCOME MEASURE: Invasive colorectal cancer incidence. RESULTS: A total of 480 incident cases of invasive colorectal cancer occurred during a mean follow-up of 8.1 (SD, 1.7) years. Intervention group participants significantly reduced their percentage of energy from fat by 10.7% more than did the comparison group at 1 year, and this difference between groups was mostly maintained (8.1% at year 6). Statistically significant increases in vegetable, fruit, and grain servings were also made. Despite these dietary changes, there was no evidence that the intervention reduced the risk of invasive colorectal cancer during the follow-up period. There were 201 women with invasive colorectal cancer (0.13% per year) in the intervention group and 279 (0.12% per year) in the comparison group (hazard ratio, 1.08; 95% confidence interval, 0.90-1.29). Secondary analyses suggested potential interactions with baseline aspirin use and combined estrogen-progestin use status (P = .01 for each). Colorectal examination rates, although not protocol defined, were comparable between the intervention and comparison groups. Similar results were seen in analyses adjusting for adherence to the intervention. CONCLUSION: In this study, a low-fat dietary pattern intervention did not reduce the risk of colorectal cancer in postmenopausal women during 8.1 years of follow-up. CLINICAL TRIALS REGISTRATION: ClinicalTrials.gov Identifier: NCT00000611. PMID- 16467234 TI - Low-fat dietary pattern and risk of cardiovascular disease: the Women's Health Initiative Randomized Controlled Dietary Modification Trial. AB - CONTEXT: Multiple epidemiologic studies and some trials have linked diet with cardiovascular disease (CVD) prevention, but long-term intervention data are needed. OBJECTIVE: To test the hypothesis that a dietary intervention, intended to be low in fat and high in vegetables, fruits, and grains to reduce cancer, would reduce CVD risk. DESIGN, SETTING, AND PARTICIPANTS: Randomized controlled trial of 48,835 postmenopausal women aged 50 to 79 years, of diverse backgrounds and ethnicities, who participated in the Women's Health Initiative Dietary Modification Trial. Women were randomly assigned to an intervention (19,541 [40%]) or comparison group (29,294 [60%]) in a free-living setting. Study enrollment occurred between 1993 and 1998 in 40 US clinical centers; mean follow up in this analysis was 8.1 years. INTERVENTION: Intensive behavior modification in group and individual sessions designed to reduce total fat intake to 20% of calories and increase intakes of vegetables/fruits to 5 servings/d and grains to at least 6 servings/d. The comparison group received diet-related education materials. MAIN OUTCOME MEASURES: Fatal and nonfatal coronary heart disease (CHD), fatal and nonfatal stroke, and CVD (composite of CHD and stroke). RESULTS: By year 6, mean fat intake decreased by 8.2% of energy intake in the intervention vs the comparison group, with small decreases in saturated (2.9%), monounsaturated (3.3%), and polyunsaturated (1.5%) fat; increases occurred in intakes of vegetables/fruits (1.1 servings/d) and grains (0.5 serving/d). Low density lipoprotein cholesterol levels, diastolic blood pressure, and factor VIIc levels were significantly reduced by 3.55 mg/dL, 0.31 mm Hg, and 4.29%, respectively; levels of high-density lipoprotein cholesterol, triglycerides, glucose, and insulin did not significantly differ in the intervention vs comparison groups. The numbers who developed CHD, stroke, and CVD (annualized incidence rates) were 1000 (0.63%), 434 (0.28%), and 1357 (0.86%) in the intervention and 1549 (0.65%), 642 (0.27%), and 2088 (0.88%) in the comparison group. The diet had no significant effects on incidence of CHD (hazard ratio [HR], 0.97; 95% confidence interval [CI], 0.90-1.06), stroke (HR, 1.02; 95% CI, 0.90-1.15), or CVD (HR, 0.98; 95% CI, 0.92-1.05). Excluding participants with baseline CVD (3.4%), the HRs (95% CIs) for CHD and stroke were 0.94 (0.86-1.02) and 1.02 (0.90-1.17), respectively. Trends toward greater reductions in CHD risk were observed in those with lower intakes of saturated fat or trans fat or higher intakes of vegetables/fruits. CONCLUSIONS: Over a mean of 8.1 years, a dietary intervention that reduced total fat intake and increased intakes of vegetables, fruits, and grains did not significantly reduce the risk of CHD, stroke, or CVD in postmenopausal women and achieved only modest effects on CVD risk factors, suggesting that more focused diet and lifestyle interventions may be needed to improve risk factors and reduce CVD risk. CLINICAL TRIALS REGISTRATION: ClinicalTrials.gov Identifier: NCT00000611. PMID- 16467235 TI - Transition of extremely low-birth-weight infants from adolescence to young adulthood: comparison with normal birth-weight controls. AB - CONTEXT: Traditionally, educational attainment, getting a job, living independently, getting married, and parenthood have been considered as markers of successful transition to adulthood. OBJECTIVE: To describe and compare the achievement and the age at attainment of the above markers between extremely low birth-weight (ELBW) and normal birth-weight (NBW) young adults. DESIGN, SETTING, AND PARTICIPANTS: A prospective, longitudinal, population-based study in central west Ontario, Canada, of 166 ELBW participants who weighed 501 to 1000 g at birth (1977-1982) and 145 sociodemographically comparable NBW participants assessed at young adulthood (22-25 years). Interviewers masked to participant status administered validated questionnaires via face-to-face interviews between January 1, 2002, and April 30, 2004. MAIN OUTCOME MEASURES: Markers of successful transition to adulthood, including educational attainment, student and/or worker role, independent living, getting married, and parenthood. RESULTS: At young adulthood, 149 (90%) of 166 ELBW participants and 133 (92%) of 145 NBW participants completed the assessments at mean (SD) age of 23.3 (1.2) years and 23.6 (1.1) years, respectively. We included participants with neurosensory impairments (ELBW vs NBW: 40 [27%] vs 3 [2%]) and 7 proxy respondents. The proportion who graduated from high school was similar (82% vs 87%, P = .21). Overall, no statistically significant differences were observed in the education achieved to date. A substantial proportion of both groups were still pursuing postsecondary education (47 [32%] vs 44 [33%]). No significant differences were observed in employment/school status; 71 (48%) ELBW vs 76 (57%) NBW young adults were permanently employed (P = .09). In a subanalysis, a higher proportion of ELBW young adults were neither employed nor in school (39 [26%] vs 20 [15%], P = .02 by Holm's correction); these differences did not persist when participants with disabilities were excluded. No significant differences were found in the proportion living independently (63 [42%] vs 70 [53%], P = .19), married/cohabitating (34 [23%] vs 33 [25%], P = .69), or who were parents (16 [11%] vs 19 [14%], P = .36). The age at attainment of the above markers was similar for both cohorts. CONCLUSION: Our study results indicate that a significant majority of former ELBW infants have overcome their earlier difficulties to become functional young adults. PMID- 16467236 TI - Comparison of two methods to detect publication bias in meta-analysis. AB - CONTEXT: Egger's regression test is often used to help detect publication bias in meta-analyses. However, the performance of this test and the usual funnel plot have been challenged particularly when the summary estimate is the natural log of the odds ratio (lnOR). OBJECTIVE: To compare the performance of Egger's regression test with a regression test based on sample size (a modification of Macaskill's test) with lnOR as the summary estimate. DESIGN: Simulation of meta analyses under a number of scenarios in the presence and absence of publication bias and between-study heterogeneity. MAIN OUTCOME MEASURES: Type I error rates (the proportion of false-positive results) for each regression test and their power to detect publication bias when it is present (the proportion of true positive results). RESULTS: Type I error rates for Egger's regression test are higher than those for the alternative regression test. The alternative regression test has the appropriate type I error rates regardless of the size of the underlying OR, the number of primary studies in the meta-analysis, and the level of between-study heterogeneity. The alternative regression test has comparable power to Egger's regression test to detect publication bias under conditions of low between-study heterogeneity. CONCLUSION: Because of appropriate type I error rates and reduction in the correlation between the lnOR and its variance, the alternative regression test can be used in place of Egger's regression test when the summary estimates are lnORs. PMID- 16467237 TI - Meeting palliative care needs in post-acute care settings: "to help them live until they die". AB - One fourth of US deaths take place in long-term care facilities. As the population ages and hospitals shorten length of stay, these settings will deliver more terminal care. Using an illustrative case of an older patient with metastatic melanoma whose life expectancy was weeks to months, we discuss when potential benefits outweigh the risks of transfer from the hospital to post-acute care settings. To improve continuity of care, we outline communication of treatment goals and orders that anticipate symptom escalation. We discuss criteria physicians can use to identify the settings most able to ensure access to high-quality palliative care. Physicians and patients must consider the advantages and disadvantages of inpatient hospice, nursing homes, and residential care facilities. Post-acute care settings vary in delivery of hospice and other palliative care services, professional nursing services, and support of activities of daily living. Finally, we discuss the evidence that palliative care can be improved in these settings, including innovations in advance care planning, staff training, and systematic changes in clinical care practices. Expanding, replicating, and disseminating these studies will be necessary to improve care for the growing number of persons who die in post-acute care settings. PMID- 16467238 TI - The automated external defibrillator: clinical benefits and legal liability. PMID- 16467239 TI - Dietary modification and risk of breast cancer. PMID- 16467240 TI - Dietary modification and CVD prevention: a matter of fat. PMID- 16467241 TI - Young adult attainments of preterm infants. PMID- 16467247 TI - JAMA patient page. Hospice care. PMID- 16467249 TI - Pharmaceutical care in community pharmacies: practice and research in Switzerland. AB - OBJECTIVE: To describe the current situation and prospects for community pharmacy practice in Switzerland, a confederation of 26 cantons with a population of approximately 7 million, located in the center of Europe. FINDINGS: For the past 10 years, the Swiss Association of Pharmacists has directed an in-depth reform of the profession, with measures such as a system of remuneration based on pharmaceutical cognitive services, a quality care program named QMS-Pharmacy, a postgraduate education program (including specific titles and certificates) and obligatory continuous education, programs of public relations and health promotion, innovative services of managed care, generics substitution, and others. DISCUSSION: The implemented changes in management represent a pharmaceutical care solution for Swiss community pharmacies, which face intense competition and a very liberal policy regarding healthcare organization, relating in particular to the important cantonal autonomy, free establishment of ownership of pharmacies, pharmacy chains, dispensing physicians, chemist shops, mail order, and others. CONCLUSIONS: Community pharmacy in Switzerland is making modernization of its role in public health policy a clear orientation. Priorities for the future include reinforcement of education and academic research in pharmacy practice, demonstration of the added value of pharmacists, integration of new information technologies with pharmacies, and development of new pharmaceutical services for managed care network in collaboration with physicians and other healthcare professionals. PMID- 16467248 TI - Chronic intrauterine pulmonary hypertension selectively modifies pulmonary artery smooth muscle cell gene expression. AB - Pulmonary artery smooth muscle cell (PASMC) relaxation at birth results from an increase in cytosolic cGMP, cGMP-dependent and kinase-mediated activation of the Ca2+-sensitive K+ channel (KCa), and closure of voltage-operated Ca2+ channels (VOCC). How chronic intrauterine pulmonary hypertension compromises perinatal pulmonary vasodilation remains unknown. We tested the hypothesis that chronic intrauterine pulmonary hypertension selectively modifies gene expression to mitigate perinatal pulmonary vasodilation mediated by the cGMP kinase-KCa-VOCC pathway. PASMC were isolated from late-gestation fetal lambs that had undergone either ligation of the ductus arteriosus (hypertensive) or sham operation (control) at 127 days of gestation and were maintained under either hypoxic (approximately 25 Torr) or normoxic (approximately 120 Torr) conditions in primary culture. We studied mRNA levels for cGMP kinase Ialpha (PKG-1alpha), the alpha-chain of VOCC (Cav1.2), and the alpha-subunit of the KCa channel. Compared with control PASMC, hypertensive PASMC had decreased VOCC, KCa, and PKG-1alpha expression. In response to sustained normoxia, expression of VOCC and KCa channel decreased and expression of PKG-1alpha increased. In contrast, sustained normoxia had no effect on PKG-1alpha levels and an attenuated effect on VOCC and KCa channel expression in hypertensive PASMC. Protein expression of PKG-1alpha was consistent with the mRNA data. We conclude that chronic intrauterine pulmonary hypertension decreases PKG expression and mitigates the genetic effects of sustained normoxia on pulmonary vasodilation, because gene expression remains compromised even after sustained exposure to normoxia. PMID- 16467250 TI - Use of aripiprazole during late pregnancy in a woman with psychotic illness. PMID- 16467251 TI - Acute respiratory failure with a single dose of quetiapine fumarate. AB - OBJECTIVE: To report a case of acute respiratory failure after a single dose of quetiapine fumarate in an elderly patient with a history of chronic obstructive pulmonary disease (COPD). CASE SUMMARY: A 92-year-old woman with a history of COPD was admitted to the hospital with pneumonia. Her symptoms improved with antibiotics. Because of acute agitation and delirium, quetiapine 50 mg twice daily was started. After receiving the first dose, the woman developed acute respiratory failure and severe central nervous system depression. She required mechanical ventilation and supportive care in the intensive care unit (ICU). She had a full recovery within 24 hours. DISCUSSION: Quetiapine is an atypical antipsychotic that has been used successfully for the treatment of schizophrenia and bipolar disorder for many years. Recently, it has also been used to treat delirium and agitation. It has proven to be very safe, even in the elderly. In previously reported cases, serious adverse effects were seen in patients who ingested very high doses of quetiapine. Those patients required intubation and supportive care in the ICU. To our knowledge, as of January 19, 2006, this is the first case report of acute respiratory failure of such severity with one dose of quetiapine. Using the Naranjo probability scale, we conclude that the acute respiratory failure observed in this patient was probably related to quetiapine. CONCLUSIONS: This case suggests that quetiapine can have significant adverse effects even with a single 50 mg dose. Elderly patients, especially those with significant underlying pulmonary pathology, should be monitored closely when started on this medication. PMID- 16467252 TI - Hormonal regulation of male reproductive tract development. AB - We have employed gene knockout technology in mice to probe gene function in various stages of mouse male sexual differentiation. Insulin-like factor (Insl3) is prominently expressed in Leydig cells. Mutation of this gene leads to fully penetrant cryptorchidism. Single mutation in each of the three known insulin family receptor tyrosine kinases alone has limited effects on sexual differentiation; however, compound mutations result in formation of ovotestes, and triple mutations cause male-to-female sexual reversal. The implications of our mouse models are discussed. PMID- 16467253 TI - Sox9 in testis determination. AB - Sox9 is an Sry-box-containing gene that encodes a transcriptional activator. During mouse gonadogenesis, Sox9 is detected in the male gonad at 11.5 days postcoitus (dpc). At 12.5 dpc, testicular cords form, morphologically distinguishing the male gonad from the ovary. From this stage onwards, Sox9 expression is restricted to the Sertoli cell lineage and persists in the adult. Humans with heterozygous mutations in SOX9 develop a skeletal syndrome known as campomelic dysplasia. Furthermore, most XY SOX9 heterozygotes show variable male to-female sex reversal, implicating SOX9 in testis development. Sox9 heterozygous knockout mice die at birth with a syndrome similar to that of human campomelic dysplasia. In contrast to humans, XY Sox9+/- mice form normal appearing testes. Germ-line knockout of Sox9 using a conditional null allele provides a tool for generating Sox9-/- mice by simple genetic crosses. However, Sox9-/- mice die soon after 11.5 dpc because of cardiovascular defects. In vitro culture of the urogenital ridges of XY Sox9-/- results in gonads lacking testicular cords and Sertoli cell marker expression, but with the expression of ovarian-specific markers. Therefore, Sox9 is essential for diverting an intrinsically ovarian program of organogenesis toward testis formation. PMID- 16467255 TI - In vitro generation of germ cells: new techniques to solve current issues. AB - Primordial germ cells comprise a privileged cellular class within the embryo charged with the elite task of maintaining species longevity. While in lower organisms germ-cell fate is determined by the allocation of germ plasm, mammalian germ-line differentiation requires extracellular signals that converge upon the proximal epiblast. Studies using mutant mice or explanted embryos have identified some of the factors controlling primordial germ-cell specification, such as members of the BMP family, but considerable gaps still exist in our understanding of the complete signaling network. Comprehensive investigations of mammalian germ line specification have been hampered by the inaccessibility of this cell population in the early embryo. Recently, however, several labs including our own have derived primordial germ cells from embryonic stem cells in vitro, thus providing a powerful new technique for the study of germ cells. In this review the different methods used for the in vitro generation of germ cells and how these techniques may be improved and applied to further advance our knowledge of germ-cell biology are discussed. PMID- 16467254 TI - Seminiferous cord formation and germ-cell programming: epigenetic transgenerational actions of endocrine disruptors. AB - The molecular and cellular control of embryonic testis development was investigated through an analysis of the embryonic testis transcriptome to identify potential regulatory factors for male sex determination and testis morphogenesis. One critical factor identified is neurotropin 3 (NT3). At the onset of male sex determination, Sertoli cells initiate differentiation and express NT3 to act as a chemotactic factor for mesonephros cells to migrate and associate with Sertoli-germ cell aggregates to promote cord formation. Promoter analysis suggests that NT3 may be an initial downstream gene to SRY and helps promote testis morphogenesis. Endocrine disruptors were used to potentially interfere with embryonic testis development and further investigate this biological process. The estrogenic pesticide methoxychlor and antiandrogenic fungicide vinclozolin were used. Previous studies have shown that methoxychlor and vinclozolin both interfere with embryonic testis cord formation and cause increased spermatogenic cell apoptosis in the adult testis. Interestingly, transient in vivo exposure to endocrine disruptors at the time of male sex determination caused a transgenerational phenotype (F1-F4) of spermatogenic cell apoptosis and subfertility. This apparent epigenetic mechanism involves altered DNA methylation and permanent re-programming of the male germ-line. A series of genes with altered DNA methylation and imprinting are being identified. Observations reviewed demonstrate that a transient embryonic in utero exposure to an endocrine disruptor influences the embryonic testis transcriptome and through epigenetic effects (e.g., DNA methylation) results in abnormal germ-cell differentiation that subsequently influences adult spermatogenic capacity and male fertility, and that this phenotype is transgenerational through the germ line. The novel observations of transgenerational epigenetic endocrine disruptor actions on male reproduction critically impact the potential hazards of these compounds as environmental toxins. The literature reviewed provides insight into the molecular and cellular control of embryonic testis development, male sex determination, and the programming of the male germ-line. PMID- 16467256 TI - Structure-function relationships of the luteinizing hormone receptor. AB - Of the 800-900 genes in the human genome that appear to encode G-protein-coupled receptors (GPCRs), two are known to encode receptors that bind the three heterodimeric human gonadotropins, luteinizing hormone (LH), chorionic gonadotropin (CG), and follicle-stimulating hormone (FSH). LH and CG bind to a common receptor, LHR, and FSH binds to a paralogous receptor. These GPCRs contain a relatively large ectodomain (ECD), responsible for high-affinity ligand binding, and a transmembrane portion, as in the other GPCRs. The ECD contains nine leucine-rich repeats capped by N-terminal and C-terminal cysteine-rich regions. The overall goal of this research is to elucidate the molecular mechanisms by which CG and LH bind to and activate LHR and the latter, in turn, activates Gs alpha. A combination of molecular modeling and site-directed mutagenesis, coupled with binding and signaling studies in transiently transfected HEK 293 cells expressing wild-type and mutant forms of LHR, has been used to develop and test models for the LHR ECD, the CG-LHR ECD complex, and the structural changes in the transmembrane helices and intracellular loops, particularly loop 2, that accompany receptor activation. In addition, a single chain CG-LHR complex was designed in which a fusion protein of the two subunits of human CG was linked to full-length LHR. This ligand-receptor complex was shown to be constitutively active in cellular models and in transgenic mice, the latter of which exhibit precocious puberty. From a combination of molecular modeling, site-directed mutagenesis, genetic/protein engineering, and receptor characterization in cellular and animal models, considerable insight is being developed on the mechanisms of normal and aberrant activation of LHR. PMID- 16467258 TI - Estrogens in testis biology. AB - Levels of estrogen within the male reproductive tract are higher than in the general circulation and the aromatase enzyme is expressed in the adult testis. Estrogens such as estradiol (E2) modify cell function by binding to high-affinity estrogen receptors (ER). Two subtypes (ERalpha and ERbeta) have been identified. Studies in animals have shown that over- or underexposure to estrogens can have an impact on testis function. For example, mice with targeted disruption of the aromatase cyp19 gene become infertile because round spermatids fail to differentiate normally. In rodents, ERalpha is expressed in Leydig cells; ERalpha mRNA and protein are not detectable in testes from humans or primates. High levels of expression of ERalpha occur in the efferent ductules in rodents, primates, and the human. ERbeta protein has been immunolocalized to all somatic cells and to some germ cells in these same species. Messenger RNAs for splice variant isoforms of human ERbeta are expressed in human testes. Homologues of the ERbeta2 variant have been cloned from primates; this isoform does not exist in rodents and does not bind E2. Full-length ERbeta protein (ERbeta1) and ERbeta2 have differential patterns of expression in human testes. In conclusion, although estrogens are synthesized in the testis and it has been suggested that E2 may function as a germ cell survival factor, the mechanisms by which estrogens influence male fertility remain uncertain and rodents may be poor models in which to examine this. PMID- 16467257 TI - Transgenic rescue of SF-1-null mice. AB - Steroidogenic factor 1 (SF-1, Nr5a1, and Ad4bp) is an orphan nuclear receptor required for adrenal and gonad development and endocrine regulation. To extend our understanding of SF-1 function and the mechanisms controlling its expression, a transgenic rescue strategy was employed to locate important transcriptional control regions and to reveal functional roles of the protein. A rat yeast artificial chromosome containing Ftz-F1, the gene encoding SF-1, was used to generate mice with different transgenes that varied in size. Rat SF-1 mRNA expression was assayed to assess each transgene's targeting ability. SF-1 deficient/transgene-positive (SF-1(-/-); tg/+) "rescue" mice were then generated and the animals' developmental and reproductive status was evaluated. The results identified differences in expression patterns and rescue abilities that provided insight into SF-1 transcriptional control and function. Comparing transgene maps and mRNA profiles placed critical transcriptional elements for pituitary and hypothalamic expression to a region 3' to intron 4, whereas examination of rescued mice revealed that an approximately 153-kb region of the Ftz-F1 locus recapitulates most or all activity ascribed to the endogenous allele. A second line of rescued mice was hypomorphic, with males showing defects in androgen dependent tissues due to abnormal Leydig cell differentiation. Histological analysis of embryonic (e14.5) and adult testes from these mice implicated SF-1 in roles that are distinct in fetal and adult Leydig cells. PMID- 16467259 TI - Foxa1 and Foxa2 interact with the androgen receptor to regulate prostate and epididymal genes differentially. AB - Previous studies from our group have shown that Foxa1 is expressed in the prostate and interacts with the androgen receptor (AR) to regulate prostate specific genes such as prostate-specific antigen (PSA) and probasin (PB). We report here that Foxa2 but not Foxa1 is expressed in the epididymis. Further, Foxa2 interacts with the AR to regulate the mouse epididymal retinoic acid binding protein (mE-RABP) gene, an epididymis-specific gene. Binding of Foxa2 to the mE-RABP promoter was confirmed by gel-shift and chromatin immunoprecipitation (ChIP) assays. Overexpression of Foxa2 suppresses androgen activation of the mE RABP promoter while overexpression of Foxa2 with prostate-specific promoters activates gene expression in an androgen-independent manner. GST pull-down assays determined that both Foxa1 and Foxa2 physically interact with the DNA binding domain of the AR. The interaction between Foxa proteins and AR was further confirmed by gel-shift assays where Foxa protein was recruited to AR binding oligomers even when Foxa binding sites were not present, and AR was recruited to Foxa binding oligomers even in the absence of an AR binding site. Given that Foxa1 and Foxa2 proteins are expressed differentially in the prostate and epididymis, these data suggest that the Foxa proteins have distinct effects on AR regulated genes in different male reproductive accessory organs. PMID- 16467260 TI - Role of glial cell line-derived neurotrophic factor in germ-line stem cell fate. AB - The overall goal of this study is to unravel the role(s) played by glial cell line-derived neurotrophic factor (GDNF) in the fate of spermatogonial stem cells. There is great interest in the biology of spermatogonial stem cells, or A(single) spermatogonia, because of their importance in the treatment of infertility, the development of contraceptives, and the understanding of the etiology of testicular cancer, particularly seminoma. In the mouse, spermatogonial stem cells express GFRalpha-1, the receptor for GDNF, and respond to this growth factor in vivo and in vitro. GDNF is produced by the adjacent Sertoli cells, which are part of the germ-line stem cell niche in vertebrates. We specifically isolated GFRalpha-1-positive spermatogonia using an immunomagnetic bead technique. We then stimulated the cells with 100 ng/mL of rGDNF for 10 hours; unstimulated cells served as negative controls. Microarray analysis, immunocytochemistry, and Western blotting revealed that Numb, a regulator of the Notch pathway, is upregulated by GDNF in spermatogonial stem cells. There are indications that in rats, mice, and humans, the Notch pathway promotes spermatogonial differentiation. We observed that an increase in Numb expression is concomitant with Notch degradation in these cells. Thus, through Numb, GDNF might inhibit differentiation and allows the maintenance of the stem cell pool in the mouse seminiferous epithelium. PMID- 16467261 TI - P450 oxidoreductase deficiency: a new disorder of steroidogenesis. AB - Microsomal P450 enzymes, which metabolize drugs and catalyze steroid biosynthesis require electron donation from NADPH via P450 oxidoreductase (POR). POR knockout mice are embryonically lethal, but we found recessive human POR missense mutations causing disordered steroidogenesis and Antley-Bixler syndrome (ABS), a skeletal malformation syndrome featuring craniosynostosis. Dominant mutations in exons 8 and 10 of fibroblast growth factor receptor 2 (FGFR2) cause phenotypically related craniosynostosis syndromes and were reported in patients with ABS and normal steroidogenesis. Sequencing POR and FGFR2 exons in 32 patients with ABS and/or hormonal findings suggesting POR deficiency showed complete genetic segregation of POR and FGFR2 mutations. Fifteen patients carried POR mutations on both alleles, four carried POR mutations on 1 allele, nine carried FGFR2/3 mutations on one allele and no mutation was found in three patients. The 34 affected POR alleles included 10 with A287P, 7 with R457H, 9 other missense mutations and 7 frameshifts. These 11 missense mutations and 10 others identified by database mining were expressed in E. coli, purified to apparent homogeneity, and their catalytic capacities were measured in four assays: reduction of cytochrome c, oxidation of NADPH, and support of the 17alpha hydroxylase and 17,20 lyase activities of human P450c17. As assessed by Vmax/Km, 17,20 lyase activity provided the best correlation with clinical findings. Modeling human POR on the X-ray crystal structure of rat POR shows that these mutant activities correlate well with their locations in the structure. POR deficiency is a new disease, distinct from the craniosynostosis syndromes caused by FGFR mutations. PMID- 16467262 TI - Neuroendocrine regulation of Leydig cell development. AB - During development in the mouse, two populations of Leydig cells arise sequentially. The fetal Leydig cell population arises shortly after testicular differentiation and functions primarily to produce androgens that are essential for masculinization of the fetus. The origin of the fetal Leydig stem cells remains uncertain, but it has been suggested that adrenocortical cells and fetal Leydig cells may share a common origin in an adrenogenital primordium. The fetal Leydig cells require an intact pituitary for normal development and are sensitive to both luteinizing hormone (LH) and adrenocorticotrophic hormone (ACTH). Loss of either one of these hormones does not, however, affect fetal androgen production, suggesting that both LH and ACTH may act to maintain fetal Leydig cell function in vivo in a redundant fashion. The adult Leydig cell population starts to develop soon after birth in the mouse. Initial differentiation does not appear to require gonadotropin input, but subsequent development and function are completely dependent upon LH. The adult Leydig cells do not require circulating follicle-stimulating hormone, provided that LH is present, but androgen stimulation, through the androgen receptor, is required for normal Leydig cell development in the mouse. It is likely that the effects of androgen are mediated directly in the Leydig cells or indirectly through the peritubular cells. PMID- 16467263 TI - Determining structure and function of steroid dehydrogenase enzymes by sequence analysis, homology modeling, and rational mutational analysis. AB - The short-chain oxidoreductase (SCOR) family of enzymes includes over 6,000 members identified in sequenced genomes. Of these enzymes, approximately 300 have been characterized functionally, and the three-dimensional crystal structures of approximately 40 have been reported. Since some SCOR enzymes are steroid dehydrogenases involved in hypertension, diabetes, breast cancer, and polycystic kidney disease, it is important to characterize the other members of the family for which the biological functions are currently unknown and to determine their three-dimensional structure and mechanism of action. Although the SCOR family appears to have only a single fully conserved residue, it was possible, using bioinformatics methods, to determine characteristic fingerprints composed of 30 40 residues that are conserved at the 70% or greater level in SCOR subgroups. These fingerprints permit reliable prediction of several important structure function features including cofactor preference, catalytic residues, and substrate specificity. Human type 1 3beta-hydroxysteroid dehydrogenase isomerase (3beta-HSDI) has 30% sequence identity with a human UDP galactose 4-epimerase (UDPGE), a SCOR family enzyme for which an X-ray structure has been reported. Both UDPGE and 3-HSDI appear to trace their origins back to bacterial 3alpha,20beta-HSD. Combining three-dimensional structural information and sequence data on the 3alpha,20beta-HSD, UDPGE, and 3beta-HSDI subfamilies with mutational analysis, we were able to identify the residues critical to the dehydrogenase function of 3-HSDI. We also identified the residues most probably responsible for the isomerase activity of 3beta-HSDI. We test our predictions by specific mutations based on sequence analysis and our structure-based model. PMID- 16467264 TI - Postnatal and pubertal development of the rhesus monkey (Macaca mulatta) testis. AB - This review examines the neurobiology, endocrinology, and cell biology underlying the development of the testis from birth until puberty in the rhesus monkey, a representative higher primate. PMID- 16467265 TI - The mRNA encoding tauCstF-64 is expressed ubiquitously in mouse tissues. AB - Polyadenylation is a process of endonucleolytic cleavage of the mRNA, followed by addition of up to 250 adenosine residues to the 3' end of the mRNA. Polyadenylation is essential for eukaryotic mRNA expression, and CstF-64 is a subunit of the CstF polyadenylation factor that is required for accurate polyadenylation. We discovered that there are two forms of the CstF-64 protein in mammalian male germ cells, one of which (CstF-64) is expressed in all tissues, the other of which (tauCstF-64) is expressed only in male germ cells and in brain (albeit at significantly lower levels in the brain). Therefore, we were surprised to find that, using reverse transcription-PCR, cDNA cloning, and RNA blot analyses, tauCstF-64 mRNA was expressed at higher levels in brain than in testis. Also, tauCstF-64 mRNA was expressed at lower but detectable levels in all tissues tested, including epididymis, heart, kidney, liver, lung, muscle, ovary, spleen, thymus, and uterus. These results suggest the hypothesis that tauCstF-64 mRNA is regulated at the translational or post-translational level. PMID- 16467266 TI - Drivers of germ cell maturation. AB - Spermatogenesis requires progression of germ line stem cells through a precisely ordered differentiation pathway to form spermatozoa. Diverse and dynamic signals from the transforming growth factor-beta (TGF-beta) superfamily influence many stages of germ cell development. For example, interactions between several TGF beta superfamily ligands (bone morphogenetic proteins, activin, and glial-derived neurotrophic growth factor [GDNF]) appear to govern the onset of spermatogenesis, and we are exploring how germ cells interpret these competing signals. We examined the in vivo impact of activin on testis development using two mouse models, the inhba-/- mouse (which lacks the gene encoding the activin A subunit and dies at birth) and BK mice, with inhbb (encoding the activin betaB subunit) replacing inhba (which survive to adulthood and show delayed fertility onset in males). Distinct effects on Sertoli cell and germ cell populations during fetal and early postnatal development were measured. We recognize that specific proteins, including downstream targets of TGF-beta signals, such as Smads, must move into the nucleus to implement the gene transcription changes required for development. We hypothesized that changes at the level of cellular nuclear transport machinery may be required to mediate this. Examination of proteins involved in classical nuclear import, the importins, revealed that each importin has a developmentally regulated expression pattern in male germ cells. Because each importin binds a selected range of cargo proteins and mediates their nucleocytoplasmic passage, our findings suggest that each importin ferries cargo required for discrete stages of spermatogenesis. PMID- 16467267 TI - Role of nuclear receptors in INSL3 gene transcription in Leydig cells. AB - Insulin-like 3 (INSL3) is a hormone produced by testicular Leydig cells throughout life. During embryonic life it regulates an essential step of testicular descent, whereas in adults it acts as a male germ cell survival factor. Despite the importance of INSL3 for male sex differentiation and function, very little is known regarding the molecular mechanisms that regulate its expression. So far, the nuclear receptor SF-1 is the only transcription factor known to regulate the mouse Insl3 promoter in Leydig cells. In order to further our understanding of the transcriptional regulation of INSL3 expression, we have isolated the human INSL3 promoter and tested the effects of the nuclear receptors SF-1, LRH-1, and Nur77 on its activity in Leydig cells. In transfections assays, all three nuclear receptors activated the human INSL3 promoter but especially Nur77, which acted through a novel regulatory element. Thus, the human INSL3 promoter constitutes a novel target for the orphan nuclear receptor Nur77. PMID- 16467268 TI - Analysis of the human sperm proteome. AB - As part of our effort to identify putative protein targets for the development of male contraceptives, we performed an in-depth proteomic analysis of human sperm by liquid chromatography and tandem mass spectrometry. Motile sperm were collected from a single fertile individual and fractionated into detergent soluble and detergent-insoluble fractions. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis separation of these fractions, followed by manual cutting of the gel, yielded 35 gel sections for each fraction to include proteins across the full range of electrophoretic mobility. Proteomic analysis of these gel sections identified more than 1,760 proteins with high confidence, with 1,350 proteins identified in the soluble fraction, 719 identified in the insoluble fraction, and 309 identified in both fractions. This characterization of the human sperm proteome provides a high-resolution, physiologically relevant index of the proteins that comprise human sperm. PMID- 16467269 TI - Male gamete contributions to the embryo. AB - During normal fertilization, plasma membranes of a spermatozoon and an oocyte mingle to form a mosaic plasma membrane of a zygote. This may contribute to the polyspermy block of the zygote. Sperm tail components (mitochondria, axonema, and accessory fibers) that enter the oocyte are "digested" without playing major roles in embryo development. The proximal centrosome adjacent to the sperm nucleus may become the center of the sperm aster that brings the male and female pronuclei to the center of the zygote, but it may not be essential for embryonic development per se. Whether sperm RNAs contribute to embryonic development is the subject of controversy. The nucleus is the most important sperm component that enters the oocyte. It is known that 7-15% of the spermatozoa of fertile men are chromosomally abnormal. The proportion of the spermatozoa with subchromosomal abnormalities (including damage at the DNA level) is expected to be even higher. The majority of embryos and fetuses with genomic abnormalities are aborted before reaching term. Structurally abnormal spermatozoa are not necessarily genomically abnormal, even though the incidence of genomically abnormal spermatozoa is higher among structurally abnormal than normal spermatozoa. In mammals, certain genes (estimated to be about 100) in the spermatozoon and oocyte must be "imprinted" in a gender-specific manner to warrant normal embryonic development. PMID- 16467270 TI - The androgen microenvironment of the human testis and hormonal control of spermatogenesis. AB - It is well established for both rat and man that the total testosterone concentration within the testis is far higher than that in serum. We know for the rat that intratesticular testosterone can be reduced by 50-60% without an adverse effect on spermatogenesis but that the required intratesticular testosterone concentration is still 10-fold greater than serum testosterone concentration. This kind of information, if available for the human, could prove invaluable for understanding and treating select men with infertility and in the development of male hormonal contraceptives. Unfortunately, we know little about the androgen content of intratesticular fluid within the human testis and nothing about the relationship between intratesticular androgens and human spermatogenesis. Using a newly developed minimally invasive technique for repetitive testicular sampling, our recent studies of the human have demonstrated that, as in the rat, there is a gradient between the concentration of testosterone in serum and within the testis; intratesticular testosterone levels were found to be 100-fold higher than serum testosterone levels in normal men. Using liquid chromatography tandem mass spectroscopy, we have shown that intratesticular 5alpha-dihydrotestosterone (DHT) levels are only 2% that of testosterone and, thus, despite greater affinity for the androgen receptor, intratesticular DHT is not significant in normal men. In order to assess how much of the testosterone within the human testis is bioactive, we adapted a highly sensitive recombinant protein mammalian cell-based bioassay to measure androgen bioactivity. The androgen bioactivity in the normal human testis is roughly two-thirds that of the total testosterone measurable by radioimmunoassay, despite the fact that the concentrations of the major androgen binding proteins (sex hormone-binding globulin- and androgen-binding protein) are insufficient to account for this difference. This finding suggests that androgens may bind to other, as-yet-unknown molecules in the human testis. How, or if, this relates to spermatogenesis in the rat, or to man-to-man differences in the response to hormonal contraceptives, is not clear. We do not yet know how much testosterone is required within the human testis to either maintain or restore quantitatively normal spermatogenesis because, as yet, experimental studies comparable to those performed in the rat have not been feasible for the human. PMID- 16467271 TI - Sperm chemotaxis: a primer. AB - Sperm become attracted by chemical substances that are released from the outer coating of the egg, a process called chemotaxis. In this paper the cellular pathway and the motor response during chemotaxis of sperm from sea urchin and starfish are briefly outlined. PMID- 16467273 TI - A journal for translational neuroscience for rehabilitation. PMID- 16467272 TI - Epigenetic regulation of testis-specific gene expression. AB - Cellular differentiation is mediated by differential gene expression. The cells of the testis are no exception. Indeed, recent studies based on microarray and expressed sequence tag analyses have revealed dynamic changes in gene expression during spermatogenesis. The autosomal phosphoglycerate kinase gene Pgk2 is an example of a gene that is tightly regulated during spermatogenesis, with transcription initiating in primary spermatocytes and ceasing in postmeiotic spermatids. Our studies show that this tissue-specific transcription is regulated at multiple levels, including binding of ubiquitous and testis-specific transcription factors, reconfiguration of nucleosomes, decondensation of chromatin, modifications of specific residues in histones, and demethylation of DNA. Our ability to isolate relatively pure populations of different spermatogenic cell types has allowed us to determine the order of these events that lead up to initiation of transcription of the Pgk2 gene. By examining similar parameters in transgenic mice carrying various portions of Pgk2 regulatory sequence, we have localized specific signals responsible for each of these regulatory events. Together these results demonstrate that activation of testis-specific transcription of the Pgk2 gene involves a multifaceted, ordered cascade of epigenetic events that potentiates this locus in preparation for the initiation of transcription. PMID- 16467274 TI - Cellular transplants in China: observational study from the largest human experiment in chronic spinal cord injury. AB - BACKGROUND: In China, fetal brain tissue has been transplanted into the lesions of more than 400 patients with spinal cord injury (SCI). Anecdotal reports have been the only basis for assuming that the procedure is safe and effective. OBJECTIVE: To compare available reports to the experiences and objective findings of patients who received pre-operative and postoperative assessments before and up to 1 year after receiving cellular implants. METHODS: Independent observational study of 7 chronic SCI subjects undergoing surgery by Dr Hongyun Huang in Beijing. Assessments included lesion location by magnetic resonance imaging, protocol of the American Spinal Injury Association (ASIA), change in disability, and detailed history of the perioperative course. RESULTS: Inclusion and exclusion criteria were not clearly defined, as subjects with myelopathies graded ASIA A through D and of diverse causes were eligible. Cell injection sites did not always correlate with the level of injury and included the frontal lobes of a subject with a high cervical lesion. Complications, including meningitis, occurred in 5 subjects. Transient postoperative hypotonicity may have accounted for some physical changes. No clinically useful sensorimotor, disability, or autonomic improvements were found. CONCLUSIONS: The phenotype and the fate of the transplanted cells, described as olfactory ensheathing cells, are unknown. Perioperative morbidity and lack of functional benefit were identified as the most serious clinical shortcomings. The procedures observed did not attempt to meet international standards for either a safety or efficacy trial. In the absence of a valid clinical trials protocol, physicians should not recommend this procedure to patients. PMID- 16467275 TI - Neuroimaging patterns associated with motor control in traumatic brain injury. AB - OBJECTIVE: To determine if patients with traumatic brain injury (TBI) and motor deficits show differences in functional activation maps during repetitive hand movements relative to healthy controls. Are there predictors for motor outcome in the functional maps of these patients? METHODS: In an exploratory cross-sectional study, functional magnetic resonance imaging (fMRI) was used to study the blood oxygenation-level-dependent (BOLD) response in cortical motor areas of 34 patients suffering from moderate motor deficits after TBI as they performed unilateral fist-clenching motions. Twelve of these patients with unilateral motor deficits were studied 3 months after TBI and a 2nd time approximately 4 months later. RESULTS: Compared to age-matched, healthy controls performing the same task, TBI patients showed diminished fMRI-signal change in the primary sensorimotor cortex contralateral to the moving hand (cSM1), the contralateral dorsal premotor cortex, and bilaterally in the supplementary motor areas (SMAs). Clinical impairment and the magnitude of the fMRI-signal change in cSM1 and SMA were negatively correlated. Patients with poor and good motor recovery showed comparable motor impairment at baseline. Only patients who evolved to "poor clinical outcome" had decreased fMRI-signal change in the cSM1 during baseline. CONCLUSIONS: These observations raise the hypothesis that the magnitude of the fMRI-signal change in the cSM1 region could have prognostic value in the evaluation of patients with TBI. PMID- 16467276 TI - Evolution of brain activation with good and poor motor recovery after stroke. AB - OBJECTIVE: To characterize the evolution of brain activation in stroke patients with variable motor recovery and quantify changes relative to healthy controls. METHODS: Serial PET activation studies, using a simple finger-tapping task, and quantitative measures of motor performance were obtained in 9 patients (2-7 weeks poststroke and 6 months later) and compared with serial healthy volunteer data. RESULTS: Patients with moderate impairment and good recovery (n = 5) activated the primary sensorimotor cortex (SM1) contralateral to the paretic hand moved, bilateral supplementary motor area (SMA), contralateral cingulate gyrus, and ipsilateral lateral premotor cortex. Activation in the bilateral SMA was greater at the initial study but reduced over time compared to healthy controls and poor recoverers. Patients with severe impairment and poor recovery (n =4) showed limited activation of contralateral SM1 and SMA at both studies and no significant change over time. A posterior shift in SM1 activation was evident in good and poor recoverers. CONCLUSIONS: Activation of typical motor regions and recruitment of additional sites occur subacutely poststroke, with evolution to normal patterns in moderately impaired patients who recover well. In comparison, severely impaired, poor-recovery patients show persistent, reduced activation. Dynamic changes in SMA, differentially observed in good recoverers over 6 months, highlight its importance in recovery. PMID- 16467277 TI - Screening patients with stroke for rehabilitation needs: validation of the post stroke rehabilitation guidelines. AB - BACKGROUND: The authors assessed patients with acute stroke to determine whether the systematic use of brief screening measures would more efficiently detect cognitive and sensory impairment than standard clinical practice. METHODS: Fifty three patients admitted to an acute stroke unit were assessed within 10 days of stroke onset. Performance on the screening measures was compared to information obtained from review of the patient's chart at discharge. Cognition, language, visual acuity, visual-spatial neglect, hearing, and depression were evaluated. RESULTS: Formal screening detected significantly more impairments than were noted in patient charts in every domain. Only 3 patients had no impairments identified on screening; all remaining patients had at least 1 impairment detected by screening that was not documented in the chart. Thirty-five percent had 3 or more undetected impairments. Memory impairment was most likely to be noted in the chart; for all other domains tested, undocumented impairment ranged from 61% (neglect) to 97% (anomia). CONCLUSION: Many acute stroke patients had cognitive and perceptual deficits that were not documented in their charts. These data support the Post-Stroke Rehabilitation Guidelines for systematic assessment even when deficits are not immediately apparent. Systematic screening may improve discharge planning, rehabilitation treatment, and long-term outcome of persons with stroke. PMID- 16467278 TI - Letter to the editor. PMID- 16467279 TI - Characterization of low density lipoprotein receptor ligand interactions by fluorescence resonance energy transfer. AB - The low density lipoprotein receptor (LDLR) is the prototype of a family of cell surface receptors involved in a wide range of biological processes. A soluble low density lipoprotein receptor (sLDLR) and a tryptophan (Trp)-deficient variant human apolipoprotein E3 (apoE3) N-terminal domain (NT) were used in binding studies. The sole cysteine in apoE3-NT was covalently modified with an extrinsic fluorescence probe, N-(iodoacetyl)-N'-(5-sulfo-1-napthyl)ethylenediamine (AEDANS), and the protein was complexed with lipid. Incubation of sLDLR with AEDANS-Trp-null apoE3-NT dimyristoylphosphatidylcholine (DMPC) disks, but not lipid-free AEDANS-apoE, induced an enhancement in AEDANS fluorescence emission intensity (excitation, 280 nm) consistent with intermolecular energy transfer from excited Trp in sLDLR to receptor-bound apoE. Ligand binding to sLDLR required calcium and was saturable. In competition binding assays, unlabeled apoE3-NT DMPC inhibited AEDANS-apoE DMPC binding to sLDLR more effectively than low density lipoprotein. Fluorescence changes in this system reflected pH dependent ligand binding and release from sLDLR consistent with models derived from the X-ray crystal structure of the receptor at endosomal pH. Intermolecular energy transfer from excited Trp in LDLR family members to fluorescently tagged ligands represents a sensitive and convenient assay for the characterization of the myriad molecular interactions ascribed to this family of receptor. PMID- 16467280 TI - Zinc Finger Protein 202, genetic variation, and HDL cholesterol in the general population. AB - Zinc Finger Protein 202 (ZNF202) is a transcriptional repressor that binds elements found predominantly in genes involved in HDL metabolism. We tested the following hypotheses: 1) frequencies of single-nucleotide polymorphisms (SNPs) and haplotypes in ZNF202 differ between individuals with low and high HDL cholesterol; and 2) SNPs in ZNF202 affect HDL cholesterol levels in the general population. We screened the promoter and protein-coding exons of ZNF202 in individuals with the highest 1% (n = 95) and lowest 1% (n = 95) HDL cholesterol among 9,259 Danish adults. None of the 10 SNPs identified differed in frequency as single sites or as haplotypes between low and high HDL cholesterol groups. In accordance with this, seven mutations were equally frequent (4-5%) in individuals with low or high HDL cholesterol. Finally, for all five SNPs identified in the coding region, we determined the association of genotype with HDL cholesterol in 9,259 individuals from the general population. Four SNPs were not associated with variation in HDL cholesterol, although c.*2T>G homozygosity was associated with a discrete effect on HDL cholesterol in men. We show that genetic variation in ZNF202 is common in the general population. However, SNPs in the protein-coding region of ZNF202 do not make a major contribution to HDL cholesterol levels. PMID- 16467281 TI - Photoprotective and anti-skin-aging effects of eicosapentaenoic acid in human skin in vivo. AB - Skin aging can be attributed to photoaging (extrinsic) and chronological (intrinsic) aging. Photoaging and intrinsic aging are induced by damage to human skin attributable to repeated exposure to ultraviolet (UV) irradiation and to the passage of time, respectively. In our previous report, eicosapentaenoic acid (EPA) was found to inhibit UV-induced matrix metalloproteinase-1 (MMP-1) expression in human dermal fibroblasts. Therefore, we investigated the effects of EPA on UV-induced skin damage and intrinsic aging by applying EPA topically to young and aged human skin, respectively. By immunohistochemical analysis and Western blotting, we found that topical application of EPA reduced UV-induced epidermal thickening and inhibited collagen decrease induced by UV light. It was also found that EPA attenuated UV-induced MMP-1 and MMP-9 expression by inhibiting UV-induced c-Jun phosphorylation, which is closely related to UV induced activator protein-1 activation, and by inhibiting JNK and p38 activation. EPA also inhibited UV-induced cyclooxygenase-2 (COX-2) expression without altering COX-1 expression. Moreover, it was found that EPA increased collagen and elastic fibers (tropoelastin and fibrillin-1) expression by increasing transformin growth factor-beta expression in aged human skin. Together, these results demonstrate that topical EPA has potential as an anti-skin-aging agent. PMID- 16467282 TI - American Academy of Pediatrics task force on sudden infant death syndrome (SIDS) statement on SIDS reduction: friend or foe of breastfeeding? PMID- 16467283 TI - Comments on "Epidural analgesia during labor and delivery" article by Chang and Heaman. PMID- 16467284 TI - The role of personality and other factors in a mother's decision to initiate breastfeeding. AB - The objective of the study was to measure the impact of personality and other factors on the decision to initiate breastfeeding. Mothers were enrolled (24-96 hours postpartum) and were classified as fully breastfeeding, formula-feeding, or combination-feeding. A semi-structured interview about maternal sociodemographics and attitudes and a standardized personality inventory (NEO-PI-R) were conducted. Eighty-seven mothers completed the study: 50 breastfeeders, 6 combination feeders, and 31 formula feeders. Because of small numbers, combination-feeder mothers (n = 6) were excluded from analyses. Maternal age, marriage, ethnicity, and socioeconomic status were significantly associated with breastfeeding. Breastfeeding and formula-feeding groups differed on 3 personality domains: extraversion (53.3 breastfeeding vs 46.9 formula-feeding, P = .002), openness (51.6 vs 46.2, P = .008), and agreeableness (48.5 vs 41.5, P = .01). In a multiple variable logistic regression model, extraversion (P = .03) and openness (P = .003) remained significant. Sociodemographics, experiential factors, and specific personality characteristics of mother were independently associated with maternal feeding decision. PMID- 16467285 TI - Barriers to compliance with infant-feeding recommendations among low-income women. AB - Focus groups were used to examine relationships among maternal beliefs, feeding intentions, and infant-feeding behaviors among 65 Women, Infants and Children eligible (28 English-speaking and 37 Spanish-speaking) mothers. Participants shared common beliefs that breast-feeding was beneficial; nevertheless, many believed that early introduction of formula and solid foods was unavoidable in certain situations. Medical providers and Women, Infants and Children staff were sources of infant-feeding information, and the Spanish-speaking mothers attempted to adhere to the guidance. However, the English-speaking mothers often ignored this advice if it was not perceived as working for the family's circumstances. Mothers, believing that providers would not understand that they were compelled to reject infant-feeding recommendations, would not ask for assistance when facing difficulties. Instead, mothers relied on relatives and others for infant feeding guidance. Educational efforts should acknowledge mothers' true circumstances, target support to each situation, and emphasize the health value of complementary foods rather than their association with infant motor development. PMID- 16467286 TI - Intragroup differences in risk factors for breastfeeding outcomes in a multicultural community. AB - A sample of 221 women who delivered at Al Qassimi Hospital, Sharjah, United Arab Emirates, were included in this prospective study to identify breastfeeding patterns at day 1, 1 month, and 6 months postpartum. The exclusive breastfeeding rate was 76.5% on day 1, 48.4% at 1 month, and 13.3% at 6 months. At 6 months, 16.1% had stopped breastfeeding. Simple and multivariable binary logistic regression analyses were used to identify factors associated with better breastfeeding outcomes. Nationality significantly affected exclusive breastfeeding at day 1 and 1 month. Pethidine use was associated with lower levels of exclusive breastfeeding at 1 month. Education was the most significant determinant of breastfeeding behavior at 6 months. Effects of the interrelationships between factors were examined and shown to influence breastfeeding outcomes in different population subgroups. The findings of this study suggest that strategies to improve breastfeeding should focus on risk factors specific to the population subgroup. PMID- 16467287 TI - Acceptability of heat treating breast milk to prevent mother-to-child transmission of human immunodeficiency virus in Zimbabwe: a qualitative study. AB - Although heat treatment of human milk is an official infant-feeding recommendation for human immunodeficiency virus (HIV)-positive mothers in Zimbabwe, its implementation has not been adequately addressed, because knowledge about the safety of this method is rudimentary and its acceptability is poorly understood. To address this knowledge gap, the authors conducted focus group discussions among mothers, grandmothers, midwives, and husbands in various regions of Zimbabwe. Although the practice of heat treating expressed human milk was initially met with skepticism because of potential obstacles, including time constraints and social and cultural stigma, a pattern of opinion reversal emerged in all groups. By the end of each discussion, participants believed that, given its affordability and its potential to protect infants from HIV infection, heat treated human milk may be a feasible infant-feeding option for HIV-positive mothers in Zimbabwe. These findings merit further investigation so that appropriate behavioral strategies can be designed. PMID- 16467288 TI - Biochemical analysis of human milk treated with sodium dodecyl sulfate, an alkyl sulfate microbicide that inactivates human immunodeficiency virus type 1. AB - Reduction of transmission of human immunodeficiency virus type 1 (HIV-1) through human milk is needed. Alkyl sulfates such as sodium dodecyl sulfate (SDS) are microbicidal against HIV-1 at low concentrations, have little to no toxicity, and are inexpensive. The authors have reported that treatment of HIV-1-infected human milk with < or = 1% (10 mg/mL) SDS for 10 minutes inactivates cell-free and cell associated virus. The SDS can be removed with a commercially available resin after treatment without recovery of viral infectivity. In this article, the authors report results of selective biochemical analyses (ie, protein, immunoglobulins, lipids, cells, and electrolytes) of human milk subjected to SDS treatment and removal. The SDS treatment or removal had no significant effects on the milk components studied. Therefore, the use of alkyl sulfate microbicides to treat milk from HIV-1-positive women may be a simple, practical, and nutritionally sound way to prevent or reduce transmission of HIV-1 while still feeding with mother's own milk. PMID- 16467289 TI - Alternative modified infant-feeding practices to prevent postnatal transmission of human immunodeficiency virus type 1 through breast milk: past, present, and future. AB - Preventing mother-to-child transmission (MTCT) of human immunodeficiency virus type 1 (HIV-1) through breastfeeding is important to reduce the number of infected children. Research on making breastfeeding safer is a high priority. The authors reviewed the attempts to develop alternative methods, other than antiretroviral (ARV) therapy of mothers and/or babies, to decontaminate breast milk of infectious HIV-1 (free and associated with lymphocytes). They also review how these methods affect milk constituents, as well as their current and prospective status. A PubMed search for English publications on methods to prevent MTCT through breast milk was completed. Methods that have been tested, other than systemicuse or ARV or immunoprophylaxis, to reduce or prevent MTCT of HIV-1 through breast milk were broadly classified into 5 groups: (1) modified feeding practices, (2) heat treatment of milk, (3) lipolysis, (4) antimicrobial treatment of the breastfeeding mother, and (5) microbicidal treatment of infected milk. Their advantages and disadvantages are discussed, as well as future directions in the prevention of MTCT through breastfeeding. PMID- 16467290 TI - Breast pump access in the inner city: a hospital-based initiative to provide breast pumps for low-income women. AB - An effective electric breast pump is an important tool for the management of breastfeeding challenges such as provision of human milk to sick or premature infants. A breast pump is also, in Western culture, critical for breastfeeding mothers who return to work. Obtaining an effective electric breast pump can be particularly difficult for uninsured or impoverished women because of the expense, complicated insurance reimbursements, and scarcity of providers that supply breast pumps to the inner-city community. To address this problem at Boston Medical Center (BMC), an inner-city hospital that serves a poor and minority urban population, members of the Breastfeeding Center worked with a local charity and local insurance companies to increase access to pumps for all women at BMC and to guarantee that every breastfeeding mother with an infant in the neonatal intensive care unit receive a double-setup electric breast pump, regardless of her insurance status or ability to pay. PMID- 16467291 TI - How to integrate a lactation consultant in an outpatient clinic environment. AB - Successful efforts in improving breastfeeding initiation rates at an urban teaching hospital prompted the hospital to create a lactation consultant (LC) position in the outpatient setting to focus on breastfeeding duration. This article reviews the complexity of the clinic setting, with the challenges and benefits of the consultant's first year in one of the hospital's outpatient clinics. Preliminary data collected by the consultant suggest that patients counseled by the LC in the outpatient clinic setting have longer breastfeeding duration rates. PMID- 16467292 TI - Designing a university-based lactation course. AB - A university-based lactation course was developed to provide lactation education to health care providers, with the goal of improving their knowledge, attitude, and skills in assisting American women to breastfeed. This is a key strategy for achieving the Healthy People 2010 objectives, and it addresses the identified need for education among health care professionals. The university as well as an enthusiastic lactation community provided multidisciplinary clinical experiences and classroom lectures to prepare public health students, nurses, midwives, and nurse practitioners to encourage and assist women in breastfeeding. It is hoped that the success of this class and the experiences of the instructors will motivate other lactation consultants to develop similar educational strategies. PMID- 16467293 TI - Reflections on the American Academy of Pediatrics' 2005 policy statement on "Breastfeeding and the use of human milk". PMID- 16467294 TI - When to see a lactation consultant. PMID- 16467296 TI - The Saccharomyces cerevisiae Lipin homolog is a Mg2+-dependent phosphatidate phosphatase enzyme. AB - Mg(2+)-dependent phosphatidate (PA) phosphatase (3-sn-phosphatidate phosphohydrolase, EC 3.1.3.4) catalyzes the dephosphorylation of PA to yield diacylglycerol and P(i). In this work, we identified the Saccharomyces cerevisiae PAH1 (previously known as SMP2) gene that encodes Mg(2+)-dependent PA phosphatase using amino acid sequence information derived from a purified preparation of the enzyme (Lin, Y.-P., and Carman, G. M. (1989) J. Biol. Chem. 264, 8641-8645). Overexpression of PAH1 in S. cerevisiae directed elevated levels of Mg(2+) dependent PA phosphatase activity, whereas the pah1Delta mutation caused reduced levels of enzyme activity. Heterologous expression of PAH1 in Escherichia coli confirmed that Pah1p is a Mg(2+)-dependent PA phosphatase enzyme and showed that its enzymological properties were very similar to those of the enzyme purified from S. cerevisiae. The PAH1-encoded enzyme activity was associated with both the membrane and cytosolic fractions of the cell, and the membrane-bound form of the enzyme was salt-extractable. Lipid analysis showed that mutants lacking PAH1 accumulated PA and had reduced amounts of diacylglycerol and its derivative triacylglycerol.ThePAH1-encoded Mg(2+)-dependent PA phosphatase shows homology to mammalian lipin, a fat-regulating protein whose molecular function is unknown. Heterologous expression of human LPIN1 in E. coli showed that lipin 1 is also a Mg(2+)-dependent PA phosphatase enzyme. PMID- 16467297 TI - Characterization of a monoclonal antibody B1 that recognizes phosphorylated Ser 158 in the activation peptide region of human coagulation factor IX. AB - Blood coagulation factor IX (FIX) undergoes various post-translational modifications such as gamma-carboxylation and glycosylation. Non-phosphorylated recombinant FIX has been reported to rapidly disappear from plasma, indicating that phosphorylation of FIX plays an important role in the physiological activity of this coagulation factor. In this study, we characterized the human FIX activation peptide (AP) using a monoclonal antibody that recognizes phosphorylated Ser-158 in the AP region. Murine monoclonal antibody B1 against human FIX recognized FIX with an apparent K(d) value of 5 nm in the presence of Ca(2+) (EC(50) = 0.58 mm). B1 bound to the isolated AP of FIX and retained the Ca(2+) dependence of binding to the isolated AP. The deglycosylation of AP did not affect the binding of B1 to AP, while B1 failed to bind to recombinant AP expressed in Escherichia coli. MALDI-TOF mass spectrometry showed that the m/z of plasma-derived deglycosylated AP is 82.54 Da greater than that of recombinant AP. The binding ability of B1 to AP was lost by the dephosphorylation of plasma derived AP. B1 bound to synthetic peptide AP-(5-19), including phosphoserine-13, but not to the non-phosphorylated AP-(5-19) in the presence of Ca(2+). These data provide direct evidence that Ser-13 of the plasma-derived FIX AP region (Ser-158 of FIX) is phosphorylated and that B1 recognizes the epitope, which includes Ca(2+)-bound phosphoserine-158. B1 should be useful in the quality control of biologically active recombinant FIX containing phosphoserine-158. PMID- 16467298 TI - AtUTr1, a UDP-glucose/UDP-galactose transporter from Arabidopsis thaliana, is located in the endoplasmic reticulum and up-regulated by the unfolded protein response. AB - The folding of glycoproteins in the endoplasmic reticulum (ER) depends on a quality control mechanism mediated by the calnexin/calreticulin cycle. During this process, continuous glucose trimming and UDP-glucose-dependent re glucosylation of unfolded glycoproteins takes place. To ensure proper folding, increases in misfolded proteins lead to up-regulation of the components involved in quality control through a process known as the unfolded protein response (UPR). Reglucosylation is catalyzed by the ER lumenal located enzyme UDP-glucose glycoprotein glucosyltransferase, but as UDP-glucose is synthesized in the cytosol, a UDP-glucose transporter is required in the calnexin/calreticulin cycle. Even though such a transporter has been hypothesized, no protein playing this role in the ER yet has been identified. Here we provide evidence that AtUTr1, a UDP-galactose/glucose transporter from Arabidopsis thaliana, responds to stimuli that trigger the UPR increasing its expression around 9-fold. The accumulation of AtUTr1 transcript is accompanied by an increase in the level of the AtUTr1 protein. Moreover, subcellular localization studies indicate that AtUTr1 is localized in the ER of plant cells. We reasoned that an impairment in AtUTr1 expression should perturb the calnexin/calreticulin cycle leading to an increase in misfolded protein and triggering the UPR. Toward that end, we analyzed an AtUTr1 insertional mutant and found an up-regulation of the ER chaperones BiP and calnexin, suggesting that these plants may be constitutively activating the UPR. Thus, we propose that in A. thaliana, AtUTr1 is the UDP glucose transporter involved in quality control in the ER. PMID- 16467299 TI - The chromosome replication machinery of the archaeon Sulfolobus solfataricus. AB - In the three domains of life, the archaea, bacteria, and eukarya, there are two general lineages of DNA replication proteins: the bacterial and the eukaryal/archaeal lineages. The hyperthermophilic archaeon Sulfolobus solfataricus provides an attractive model for biochemical study of DNA replication. Its relative simplicity in both genomic and biochemical contexts, together with high protein thermostability, has already provided insight into the function of the more complex yet homologous molecules of the eukaryotic domain. Here, we provide an overview of recent insights into the functioning of the chromosome replication machinery of S. solfataricus, focusing on some of the relatively well characterized core components that act at the DNA replication fork. PMID- 16467300 TI - Expression cloning and periplasmic orientation of the Francisella novicida lipid A 4'-phosphatase LpxF. AB - Francisella tularensis and related intracellular pathogens synthesize lipid A molecules that differ from their Escherichia coli counterparts. Although a functional orthologue of lpxK, the gene encoding the lipid A 4'-kinase, is present in Francisella, no 4'-phosphate moiety is attached to Francisella lipid A. We now demonstrate that a membrane-bound phosphatase present in Francisella novicida U112 selectively removes the 4'-phosphate residue from tetra- and pentaacylated lipid A molecules. A clone that expresses the F. novicida 4' phosphatase was identified by assaying lysates of E. coli colonies, harboring members of an F. novicida genomic DNA library, for 4'-phosphatase activity. Sequencing of a 2.5-kb F. novicida DNA insert from an active clone located the structural gene for the 4'-phosphatase, designated lpxF. It encodes a protein of 222 amino acid residues with six predicted membrane-spanning segments. Rhizobium leguminosarum and Rhizobium etli contain functional lpxF orthologues, consistent with their lipid A structures. When F. novicida LpxF is expressed in an E. coli LpxM mutant, a strain that synthesizes pentaacylated lipid A, over 90% of the lipid A molecules are dephosphorylated at the 4'-position. Expression of LpxF in wild-type E. coli has no effect, because wild-type hexaacylated lipid A is not a substrate. However, newly synthesized lipid A is not dephosphorylated in LpxM mutants by LpxF when the MsbA flippase is inactivated, indicating that LpxF faces the outer surface of the inner membrane. The availability of the lpxF gene will facilitate re-engineering lipid A structures in diverse bacteria. PMID- 16467301 TI - Cys-113 and Cys-422 form a high affinity metalloid binding site in the ArsA ATPase. AB - The arsRDABC operon of Escherichia coli plasmid R773 encodes the ArsAB extrusion pump for the trivalent metalloids As(III) and Sb(III). ArsA, the catalytic subunit has two homologous halves, A1 and A2. Each half has a consensus signal transduction domain that physically connects the nucleotide-binding domain to the metalloid-binding domain. The relation between metalloid binding by ArsA and transport through ArsB is unclear. In this study, direct metalloid binding to ArsA was examined. The results show that ArsA binds a single Sb(III) with high affinity only in the presence of Mg(2+)-nucleotide. Mutation of the codons for Cys-113 and Cys-422 eliminated Sb(III) binding to purified ArsA. C113A/C422A ArsA has basal ATPase activity similar to that of the wild type but lacks metalloid stimulated activity. Accumulation of metalloid was assayed in intact cells, where reduced uptake results from active extrusion by the ArsAB pump. Cells expressing the arsA(C113A/C422A)B genes had an intermediate level of metalloid resistance and accumulation between those expressing only arsB alone and those expressing wild type arsAB genes. The results indicate that, whereas metalloid stimulation of ArsA activity enhances the ability of the pump to reduce the intracellular concentration of metalloid, high affinity binding of metalloid by ArsA is not obligatory for transport or resistance. Yet, in mixed populations of cells bearing either arsAB or arsA(C113A/C422A)B growing in subtoxic concentrations of arsenite, cells bearing wild type arsAB replaced cells with mutant arsA(C113A/C422A)B in less than 1 week, showing that the metalloid binding site confers an evolutionary advantage. PMID- 16467302 TI - Thermal stabilization of Bacillus subtilis family-11 xylanase by directed evolution. AB - We used directed evolution to enhance the thermostability of glycosyl hydrolase family-11 xylanase from Bacillus subtilis. By combining random point mutagenesis, saturation mutagenesis, and DNA shuffling, a thermostable variant, Xyl(st), was identified which contained three amino acid substitutions: Q7H, N8F, and S179C. The half-inactivation temperature (the midpoint of the melting curves) for the Xyl(st) variant compared with the wild-type enzyme after incubation for 10 min was elevated from 58 to 68 degrees C. At 60 degrees C the wild-type enzyme was inactivated within 5 min, but Xyl(st) retained full activity for at least 2 h. The stabilization was accompanied by evidence of thermophilicity; that is, an increase in the optimal reaction temperature from 55 to 65 degrees C and lower activity at low temperatures and higher activity at higher temperatures relative to wild type. To elucidate the mechanism of thermal stabilization, three dimensional structures were determined for the wild-type and Xyl(st) enzymes. A cavity was identified around Gln-7/Asn-8 in wild type that was filled with bulky, hydrophobic residues in Xyl(st). This site was not identified by previous approaches, but directed evolution identified the region as a weak point. Formation of an intermolecular disulfide bridge via Cys-179 was observed between monomers in Xyl(st). However, the stability was essentially the same in the presence and absence of a reducing agent, indicating that the increased hydrophobicity around the Cys-179 accounted for the stability. PMID- 16467303 TI - RNA aptamers targeting the cell death inhibitor CED-9 induce cell killing in Caenorhabditis elegans. AB - Bcl-2 family proteins include anti- and proapoptotic factors that play important roles in regulating apoptosis in diverse species. Identification of compounds that can modulate the activities of Bcl-2 family proteins will facilitate development of drugs for treatment of apoptosis-related human diseases. We used an in vitro selection method named systematic evolution of ligands by exponential enrichment (SELEX) to isolate RNA aptamers that bind the Caenorhabditis elegans Bcl-2 homolog CED-9 with high affinity and specificity and tested whether these aptamers modulate programmed cell death in C. elegans. Five CED-9 aptamers were isolated and classified into three groups based on their predicted secondary structures. Biochemical analyses indicated that two of these aptamers, R9-2 and R9-7, and EGL-1, an endogenous CED-9-binding proapoptotic protein, bound to distinct regions of CED-9. However, these two aptamers shared overlapping CED-9 binding sites with CED-4, another CED-9-binding proapoptotic factor. Importantly ectopic expression of these two aptamers in touch receptor neurons induced efficient killing of these neurons largely in a CED-3 caspase-dependent manner. These findings suggest that RNA aptamers can be used to modulate programmed cell death in vivo and can potentially be used to develop drugs to treat human diseases caused by abnormal apoptosis. PMID- 16467304 TI - Lipid phosphate phosphatase-2 activity regulates S-phase entry of the cell cycle in Rat2 fibroblasts. AB - Lipid phosphates are potent mediators of cell signaling and control processes including development, cell migration and division, blood vessel formation, wound repair, and tumor progression. Lipid phosphate phosphatases (LPPs) regulate the dephosphorylation of lipid phosphates, thus modulating their signals and producing new bioactive compounds both at the cell surface and in intracellular compartments. Knock-down of endogenous LPP2 in fibroblasts delayed cyclin A accumulation and entry into S-phase of the cell cycle. Conversely, overexpression of LPP2, but not a catalytically inactive mutant, caused premature S-phase entry, accompanied by premature cyclin A accumulation. At high passage, many LPP2 overexpressing cells arrested in G(2)/M and the rate of proliferation declined severely. This was accompanied by changes in proteins and lipids characteristic of senescence. Additionally, arrested LPP2 cells contained decreased lysophosphatidate concentrations and increased ceramide. These effects of LPP2 activity were not reproduced by overexpression or knock-down of LPP1 or LPP3. This work identifies a novel and specific role for LPP2 activity and bioactive lipids in regulating cell cycle progression. PMID- 16467305 TI - Crystal structure of the V domain of human Nectin-like molecule 1/Syncam3/Tsll1/Igsf4b, a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule. AB - Nectins are Ca(2+)-independent immunoglobulin (Ig) superfamily proteins that participate in the organization of epithelial and endothelial junctions. Nectins have three Ig-like domains in the extracellular region, and the first one is essential in cell-cell adhesion and plays a central role in the interaction with the envelope glycoprotein D of several viruses. Five Nectin-like molecules (Necl 1 through -5) with similar domain structures to those of Nectins have been identified. Necl-1 is specifically expressed in neural tissue, has Ca(2+) independent homophilic and heterophilic cell-cell adhesion activity, and plays an important role in the formation of synapses, axon bundles, and myelinated axons. Here we report the first crystal structure of its N-terminal Ig-like V domain at 2.4 A, providing insight into trans-cellular recognition mediated by Necl-1. The protein crystallized as a dimer, and the dimeric form was confirmed by size exclusion chromatography and chemical cross-linking experiments, indicating this V domain is sufficient for homophilic interaction. Mutagenesis work demonstrated that Phe(82) is a key residue for the adhesion activity of Necl-1. A model for homophilic adhesion of Necl-1 at synapses is proposed based on its structure and previous studies. PMID- 16467306 TI - Mutational analysis of membrane and soluble forms of human MD-2. AB - Toll-like receptor 4 and MD-2 form a receptor for lipopolysaccharide (LPS), a major constituent of Gram-negative bacteria. MD-2 is a 20-25-kDa extracellular glycoprotein that binds to Tolllike receptor 4 (TLR4) and LPS and is a critical part of the LPS receptor. Here we have shown that the level of MD-2 expression regulates TLR4 activation by LPS. Using site-directed mutagenesis, we have found that glycosylation has no effect on MD-2 function as a membrane receptor for LPS. We used alanine-scanning mutagenesis to identify regions of human MD-2 that are important for TLR4 and LPS binding. We found that mutation in the N-terminal 46 amino acids of MD-2 did not substantially diminish LPS activation of Chinese hamster ovary (CHO) cells co-transfected with TLR4 and mutant MD-2. The residues 46-50 were important for LPS activation but not LPS binding. The residues 79-83, 121-124, and 125-129 are identified as important in LPS activation but not surface expression of membrane MD-2. The function of soluble MD-2 is somewhat more sensitive to mutation than membrane MD-2. Our results suggest that the 46-50 and 127-131 regions of soluble MD-2 bind to TLR4. The region 79-120 is not involved in LPS binding but affects monomerization of soluble MD-2 as well as TLR4 binding. We define the LPS binding region of monomeric soluble MD-2 as a cluster of basic residues 125-131. Studies on both membrane and soluble MD-2 suggest that domains of MD-2 for TLR4 and LPS binding are separate as well as overlapping. By mapping these regions on a three-dimensional model, we show the likely binding regions of MD-2 to TLR4 and LPS. PMID- 16467307 TI - Kinetics and thermodynamics of ligand binding by cytochrome P450 3A4. AB - Cytochrome P450 (P450) 3A4, the major catalyst involved in human drug oxidation, displays substrate- and reaction-dependent homotropic and heterotropic cooperative behavior. Although several models have been proposed, these mainly rely on steady-state kinetics and do not provide information on the contribution of the individual steps of P450 catalytic cycle to the observed cooperativity. In this work, we focused on the kinetics of substrate binding, and the fluorescent properties of bromocriptine and alpha-naphthoflavone allowed analysis of an initial ligand-P450 3A4 interaction that does not cause a perturbation of the heme spectrum. The binding stoichiometry for bromocriptine was determined to be unity using isothermal titration calorimetry and equilibrium dialysis methods, suggesting that the ligand bound to the peripheral site during the initial encounter dissociates subsequently. A three-step substrate binding model is proposed, based on absorbance and fluorescence stopped-flow kinetic data and equilibrium binding data obtained with bromocriptine, and evaluated using kinetic modeling. The results are consistent with the substrate molecule binding at a site peripheral to the active site and subsequently moving toward the active site to bind to the heme and resulting in a low to high spin iron shift. The last step is attributed to a conformational change in the enzyme active site. The later steps of binding were shown to have rate constants comparable with the subsequent steps of the catalytic cycle. The P450 3A4 binding process is more complex than a two-state system, and the overlap of rates of some of the events with subsequent steps is proposed to underlie the observed cooperativity. PMID- 16467308 TI - Complex role of the vitamin D receptor and its ligand in adipogenesis in 3T3-L1 cells. AB - The vitamin D receptor (VDR) and its ligand 1,25-OH2-VD3 (calcitriol) play an essential role in mineral homeostasis in mammals. Interestingly, the VDR is expressed very early in adipogenesis in 3T3-L1 cells, suggesting that the VDR signaling pathway may play a role in adipocyte biology and function. Indeed, it has been known for a number of years that calcitriol is a potent inhibitor of adipogenesis in this model but with no clear mechanism identified. In this study, we have further defined the molecular mechanism by which the unliganded VDR and calcitriol-liganded VDR regulate adipogenesis. In the presence of calcitriol, the VDR blocks adipogenesis by down-regulating both C/EBPbeta mRNA expression and C/EBPbeta nuclear protein levels at a critical stage of differentiation. In addition, calcitriol allows for the up-regulation of the recently described C/EBPbeta corerepressor, ETO, which would further inhibit the action of any remaining C/EBPbeta, whose action is required for adipogenesis. In contrast, in the absence of calcitriol, the unliganded VDR appears necessary for lipid accumulation, since knock-down of the VDR using siRNA both delays and prevents this process. Taken together, these data support the notion that the intracellular concentrations of calcitriol can play an important role in either promoting or inhibiting adipogenesis via the VDR and the transcriptional pathways that it targets. Further examination of this hypothesis in vivo may shed new light on the biology of adipogenesis. PMID- 16467310 TI - Recombinant expression and biochemical characterization of the unique elongating beta-ketoacyl-acyl carrier protein synthase involved in fatty acid biosynthesis of Plasmodium falciparum using natural and artificial substrates. AB - The human malaria parasite Plasmodium falciparum synthesizes fatty acids by using a type II synthase that is structurally different from the type I system found in eukaryotes. Because of this difference and the vital role of fatty acids, the enzymes involved in fatty acid biosynthesis of P. falciparum represent interesting targets for the development of new antimalarial drugs. beta-Ketoacyl acyl carrier protein (ACP) synthase (PfFabBF), being the only elongating beta ketoacyl-ACP synthase in P. falciparum, is a potential candidate for inhibition. In this study we present the cloning, expression, purification, and characterization of PfFabBF. Soluble protein was obtained when PfFabBF was expressed as a NusA fusion protein in Escherichia coli BL21(DE3)-CodonPlus-RIL cells under conditions of osmotic stress. The fusion protein was purified by affinity and ion exchange chromatography. Various acyl-P. falciparum acyl carrier protein (PfACP) substrates were tested for their specific activities, and their kinetic parameters were determined. Activity of PfFabBF was highest with C(4:0)- to C(10:0)-acyl-PfACPs and decreased with use of longer chain acyl-PfACPs. Consistent with the fatty acid synthesis profile found in the parasite cell, no activity could be detected with C(16:0)-PfACP, indicating that the enzyme is lacking the capability of elongating acyl chains that are longer than 14 carbon atoms. PfFabBF was found to be specific for acyl-PfACPs, and it displayed much lower activities with the corresponding acyl-CoAs. Furthermore, PfFabBF was shown to be sensitive to cerulenin and thiolactomycin, known inhibitors of beta ketoacyl-ACP synthases. These results represent an important step toward the evaluation of P. falciparum beta-ketoacyl-ACP synthase as a novel antimalaria target. PMID- 16467309 TI - Cyclooxygenase-2 induction and prostacyclin release by protease-activated receptors in endothelial cells require cooperation between mitogen-activated protein kinase and NF-kappaB pathways. AB - The functional significance of protease-activated receptors (PARs) in endothelial cells is largely undefined, and the intracellular consequences of their activation are poorly understood. Here, we show that the serine protease thrombin, a PAR-1-selective peptide (TFLLRN), and SLIGKV (PAR-2-selective peptide) induce cyclooxygenase-2 (COX-2) protein and mRNA expression in human endothelial cells without modifying COX-1 expression. COX-2 induction was accompanied by sustained production of 6-keto-PGF1alpha, the stable hydrolysis product of prostacyclin, and this was inhibited by indomethacin and the COX-2 selective inhibitor NS398. PAR-1 and PAR-2 stimulation rapidly activated both ERK1/2 and p38MAPK, and pharmacological blockade of MEK with either PD98059 or U0126 or of p38MAPK by SB203580 or SB202190 strongly inhibited thrombin- and SLIGKV-induced COX-2 expression and 6-keto-PGF1alpha formation. Thrombin and peptide agonists of PAR-1 and PAR-2 increased luciferase activity in human umbilical vein endothelial cells infected with an NF-kappaB-dependent luciferase reporter adenovirus, and this, as well as PAR-induced 6-keto-PGF1alpha synthesis, was inhibited by co-infection with adenovirus encoding wild-type or mutated (Y42F) IkappaBalpha. Thrombin- and SLIGKV-induced COX-2 expression and 6-keto PGF1alpha generation were markedly attenuated by the NF-kappaB inhibitor PG490 and partially inhibited by the proteasome pathway inhibitor MG-132. Activation of PAR-1 or PAR-2 promoted nuclear translocation and phosphorylation of p65-NF kappaB, and thrombin-induced but not PAR-2-induced p65-NF-kappaB phosphorylation was reduced by inhibition of MEK or p38MAPK. Activation of PAR-4 by AYPGKF increased phosphorylation of ERK1/2 and p38MAPK without modifying NF-kappaB activation or COX-2 induction. Our data show that PAR-1 and PAR-2, but not PAR-4, are coupled with COX-2 expression and sustained endothelial production of vasculoprotective prostacyclin by mechanisms that depend on ERK1/2, p38MAPK, and IkappaBalpha-dependent NF-kappaB activation. PMID- 16467311 TI - Sleep in overweight adolescents: shorter sleep, poorer sleep quality, sleepiness, and sleep-disordered breathing. AB - OBJECTIVE: To document the sleep of overweight adolescents and to explore the degree to which weight-related sleep pathology might account for diminished psychosocial outcome. METHODS: Sixty children aged 10-16.9 from a weight management clinic were compared to 22 healthy controls using comprehensive actigraphic, polysomnographic, and parent- and self-report questionnaire assessments. RESULTS: Overweight participants averaged more symptoms of sleep disordered breathing, later sleep onset, shorter sleep time, and more disrupted sleep than controls. Although the groups did not differ in self-reported sleep habits, multiple concerns were reported by parents of overweight participants, including daytime sleepiness, parasomnias, and inadequate sleep. Group differences in academic grades and depressive symptoms were at least partially accounted for by short sleep and daytime sleepiness. CONCLUSIONS: Excessive weight is associated with an increased risk of sleep problems. There is a need for further research in this area and for clinicians who work with overweight children to evaluate their sleep. PMID- 16467312 TI - Introduction to the special section on families, youth, and HIV: family-based intervention studies. PMID- 16467313 TI - Factor structure of the child health questionnaire-parent form in pediatric populations. AB - OBJECTIVE: To conduct separate exploratory factor analyses (EFA) and confirmatory factor analyses (CFA) of the Child Health Questionnaire-Parent Form 50 (CHQ-PF 50) with a sample of children and adolescents with chronic conditions and physically healthy children seen in a pediatric setting. METHOD: Parents of 329 children with chronic conditions including cancer, epilepsy, recurrent headache, inflammatory bowel disease (IBD), juvenile rheumatoid arthritis (JRA), sickle cell disease (SCD), and recurrent sleep disturbance and 332 physically healthy children completed CHQ-PF-50. RESULTS: The EFA yielded a 27-item measure with seven factors for children with chronic conditions and a 28-item measure with eight factors for physically healthy children. Structural equation modeling procedures were used to conduct a second order CFA, which yielded the secondary factors of physical health and psychosocial health. A CFA yielded an excellent fit to the data for each group, but the models were different for each group. CONCLUSIONS: CFA-derived models of the CHQ-PF-50 demonstrated construct validity for measuring the latent constructs of physical and psychosocial health in children and adolescents with chronic conditions and physically healthy children and adolescents. However, somewhat different factor solutions emerged for each group, suggesting that the specific domains assessed by the CHQ-PF-50 were not equivalent across groups. Findings have implications for applications of the CHQ PF-50. PMID- 16467314 TI - The impact of self-control indices on peer smoking and adolescent smoking progression. AB - OBJECTIVE: To determine the direct impact of self-control variables on baseline smoking and smoking progression and determine whether self-control had indirect effects on smoking practices through effects on peer smoking. METHODS: Study participants were 918 adolescents who were followed from 9th through the 12th grade and completed self-report measures of peer smoking, self-control, and cigarette smoking. An exploratory factor analysis (EFA) was conducted to assess the factor structure of a 41-item self-control measure. The EFA indicated a six factor structure comprising of impulsive control, planning, hostile blaming, attentional disregulation, conscientiousness, and physical aggression. RESULTS: The results of a latent growth model indicated that conscientiousness (OR = 0.81, CI = 0.73-0.90), hostile blaming (OR = 0.89, CI = 0.81-0.99), and physical aggression (OR = 1.16, CI = 1.06-1.27) had direct effects on baseline smoking, whereas planning (OR = 0.90, CI = 0.82-0.99) and impulse control (OR = 1.15, CI = 1.02-1.28) had indirect effects on adolescent smoking at baseline through baseline peer smoking. There were no significant direct or indirect effects of the self-control indices on smoking progression. There was a direct effect of peer smoking progression (number of peers who smoked) on adolescent smoking progression, such that increases in the number of peers who smoked across time increased the odds that an adolescent would progress to a higher level of smoking. CONCLUSIONS: Youth smoking prevention and intervention program outcomes may potentially improve by addressing self-control behaviors as they appear to have direct effects on smoking and indirect effects through peers who smoke. PMID- 16467315 TI - The Stamp-in-Safety program: a behavioral intervention to reduce behaviors that can lead to unintentional playground injury in a preschool setting. AB - OBJECTIVE: To introduce and evaluate the Stamp-in-Safety program, a behavioral intervention designed to increase the quality of supervision by teachers on preschool playgrounds and to reduce the risk of unintentional child playground injury. METHODS: A quasi-experimental time series design was used, with observational techniques, to evaluate behavior on the playground before, during, and after the intervention as well as during a 6-month post-intervention assessment. RESULTS: Both applied behavior analysis techniques and inferential statistics suggest that the Stamp-in-Safety program resulted in behavioral changes likely to reduce the risk of child injury on the playgrounds of childcare centers. CONCLUSION: Results indicate promise for the Stamp-in-Safety program as a low-cost, easily implemented intervention to reduce pediatric playground injury risk at childcare centers. Suggestions for future research are offered. PMID- 16467316 TI - A longitudinal examination predicting emergency room use in children with sickle cell disease and their caregivers. AB - OBJECTIVE: To examine in a longitudinal investigation perceptions of disease severity among children and adolescents with sickle cell disease (SCD) and their caregivers and to determine whether perceptions of disease severity and self reported adjustment would account for a greater percentage of the variance in emergency room (ER) use than objective measures of disease severity. METHODS: Participants were 58 African-American children (ages 8-18 years) diagnosed with SCD who were recruited from an outpatient clinic. Children and their caregivers were administered a series of measures designed to assess caregivers' adjustment and perceptions of children's disease severity. Frequency of ER visits was tabulated over an average 4-year period. RESULTS: Caregivers' and children's social adjustment accounted for a unique portion of the variance beyond that accounted by subjective perceptions of disease severity, demographic, and objective biological markers of disease severity. Poorer psychological adjustment of caregivers as well as communication patterns among children were associated with ER use frequency after controlling for objective disease severity variables and demographic factors. Child social adjustment, specifically friendship quality, accounted for variance in children's use of the ER beyond that accounted for by social class and objective markers of disease severity. CONCLUSIONS: Support is provided for the notion that ER use for children with SCD may be independent of disease severity and predicted by caregivers' psychological adjustment and children's peer adjustment. PMID- 16467317 TI - The relation between family factors and metabolic control: the role of diabetes adherence. AB - OBJECTIVES: To examine family factors as predictors of metabolic control in children with type 1 diabetes and determine whether adherence behaviors mediate this relationship. METHOD: Participants were 109 children (ages 8-18) and a parent. Measures of diabetes-specific family functioning and an adherence interview were completed. Glycosylated hemoglobin (HbA1c) was the index of metabolic control. RESULTS: Family functioning and adherence were strongly associated with metabolic control. Combined with demographic information, these constructs accounted for 49% of the variance in metabolic control. Age moderated the relation between aspects of family functioning and HbA1c. Path analyses suggest that adherence mediates the relationship between family functioning and metabolic control. CONCLUSIONS: Family functioning and adherence behaviors are strongly related to a child's health status. Assessment of diabetes-specific family functioning, in addition to adherence, is an important factor in understanding metabolic control. PMID- 16467318 TI - Does time spent in child care influence risk for unintentional injury? AB - OBJECTIVE: To investigate the effect of quantity and quality of early child care on children's risk for unintentional injury. METHODS: A diverse cohort of 1,225 children was recruited from several sites in the United States and followed from birth until first grade. Quantity and quality of child care from birth until entry into kindergarten were used to predict unintentional injuries from age 6 months until first grade. Measures from an evaluation at 6 months of age were tested as covariates. RESULTS: Children who spent more time in nonparental childcare environments were at slightly reduced risk for unintentional injury after controlling for child (gender, temperament), family [socioeconomic status (SES)], parent (positive parenting), and child care (quality of care) characteristics. CONCLUSIONS: We discuss possible explanations for the results, including the possibilities that childcare center environments are safer than the homes of most preschoolers or that attendance in child care is nonrandom. PMID- 16467319 TI - Brief report: friendships of adolescents with and without diabetes. AB - BACKGROUND: Friendships of adolescents with chronic illness have been rarely studied. OBJECTIVE: To compare the friendships of boys and girls with diabetes with those between healthy adolescents. METHODS: One hundred and thirty-eight adolescents were interviewed at summer camps. Participants indicated their number of close friends and rated friendships for support and conflict. Participants described aspects of their same-sex and other-sex friendships that they liked and disliked. RESULTS: Adolescents with diabetes have friendships that are similar to those between healthy adolescents, with a few exceptions. CONCLUSIONS: Emotional support may be especially valued by girls with diabetes, whereas differences among friends may be less valued among adolescents with diabetes. PMID- 16467320 TI - Factor analysis and validity of the Conners Parent and Teacher Rating Scales in childhood cancer survivors. AB - OBJECTIVE: To examine the factor structure of the Conners Parent Rating Scale Revised: Short Form (CPRS-R:S) and the Conners Teacher Rating Scale-Revised: Short Form (CTRS-R:S) in children who are long-term survivors of acute lymphocytic leukemia (ALL) or brain tumors (BT)and who have received central nervous system directed treatment. METHOD: Parents and teachers of 150 long-term survivors completed the CPRS-R:S or CTRS-R:S as part of a screening battery. The data were submitted to a maximum likelihood confirmatory factor analysis to test the construct validity of the scales and the forms were compared. The CPRS-R:S was also compared to selected subscales of the Achenbach Child Behavior Checklist (CBCL) for further validation. RESULTS: The analyses demonstrated an adequate fit of the original three-factor structure of the CTRS-R:S [oppositional, cognitive problems/inattention, hyperactivity]. The analyses of the CPRS-R:S suggested a less adequate fit of the original three-factor structure but principal components factor analysis yielded a three-factor solution with factors similar to those of Conners' original factor structure. Significant correlations were found between the CPRS-R:S and the selected subscales of the CBCL. CONCLUSIONS: These findings support the similar construct validity of the original CTRS-R:S and CPRS-R:S. Although significantly correlated, the CPRS-R:S and CTRS-R:S are not interchangeable in the assessment of survivors of childhood cancer. PMID- 16467321 TI - Use of opsonophagocytosis for serological evaluation of pneumococcal vaccines. PMID- 16467322 TI - Whipple's disease: a macrophage disease. PMID- 16467324 TI - Performance of antibodies against tissue transglutaminase for the diagnosis of celiac disease: meta-analysis. AB - A meta-analysis of studies investigating the diagnostic accuracy of enzyme-linked immunosorbent assays (ELISA) for antibodies against tissue transglutaminases (tTG) of various origins in celiac disease (CD) diagnosis was carried out. Twenty one studies, with untreated CD patients and healthy/CD-free controls, were included in the meta-analysis. The diagnostic accuracy was estimated using a summary receiver operating characteristic (SROC) curve and pooled sensitivity (Se) and specificity (Sp). Multiple assays within a study were treated by considering all the assays within a study and by analyzing the most popular assay (i.e., the commercial anti-tTTG ELISA most frequently utilized in the papers in which multiple assays were included). The SROC curve indicated the absence of heterogeneity, and the superiority of recombinant human tTG (rh-tTG) and purified human tTG (ph-tTG) compared to guinea pig-tTG (gp-tTG). The sensitivities (most popular assay) for rh-tTG, ph-tTG, and gp-tTG were 94%, 90%, and 92%, respectively, and the specificities were 97%, 92%, and 96%, respectively. A sensitivity analysis (exclusion of studies with bias) altered the results of ph tTG: Se, 95%; Sp, 98%. The sensitivities (all individual assays) for rh-tTG, ph tTG, and gp-tTG were 94%, 94%, and 91%, respectively, and the specificities were 95%, 94%, and 89%, respectively. Human tTG ELISA is sensitive and specific, and it can be used for mass screening. Sensitivity analysis showed that ph-tTG might perform better. PMID- 16467323 TI - Recognition of stage-specific mycobacterial antigens differentiates between acute and latent infections with Mycobacterium tuberculosis. AB - Mycobacterium tuberculosis is estimated to infect 80 to 100 million people annually, the majority of whom do not develop clinical tuberculosis (TB) but instead maintain the infection in a latent state. These individuals generally become positive in response to a tuberculin skin test and may develop clinical TB at a later date, particularly if their immune systems are compromised. Latently infected individuals are interesting for two reasons. First, they are an important reservoir of M. tuberculosis, which needs to be considered for TB control. Second, if detected prior to recrudescence of the disease, they represent a human population that is making a protective immune response to M. tuberculosis, which is very important for defining correlates of protective immunity. In this study, we show that while responsiveness to early secretory antigenic target 6 is a good marker for M. tuberculosis infection, a strong response to the 16-kDa Rv2031c antigen (HspX or alpha-crystallin) is largely restricted to latently infected individuals, offering the possibility of differential immunodiagnosis of, or therapeutic vaccination against, TB. PMID- 16467325 TI - Combination exposure to zidovudine plus sulfamethoxazole-trimethoprim diminishes B-lymphocyte immune responses to Pneumocystis murina infection in healthy mice. AB - We have previously shown that zidovudine plus sulfamethoxazole-trimethoprim exposure decreases immune cell populations in the bone marrow of healthy mice by inducing apoptosis. The hypothesis of the current work was that this toxicity would have an adverse impact on the immune response. To determine this, BALB/c mice were treated with zidovudine, sulfamethoxazole-trimethoprim, the combination of both drugs, or vehicle only (control) via oral gavage for 21 days. On day 4 after dosing completion, the mice were infected intratracheally with 1x10(7) Pneumocystis murina organisms. Immune cell populations (in lung digest, bronchoalveolar lavage fluid, tracheobronchial lymph node, and bone marrow samples), the lung Pneumocystis burden, and serum Pneumocystis-specific antibody titers were determined at days 6, 10, and 20 postinfection. While total bone marrow cellularity was recovered by day 6 postinfection in the combination exposure group, B-cell numbers did not recover until 10 days postinfection, primarily due to the persistent depletion of the late pre-B-cell phenotype. The numbers of CD4+ and CD8+ T cells, as well as the numbers of total B cells and activated B cells in tracheobronchial lymph nodes, were decreased at days 10 and 20 as a result of zidovudine plus sulfamethoxazole-trimethoprim exposure compared to the numbers in the control group. No significant differences in lung lavage or lung digest cell populations were observed. There was a trend of a delay in Pneumocystis clearance in the combination treatment group, and Pneumocystis specific serum immunoglobulin G titers were reduced at day 20 postinfection. Together, these data indicate that the combination of zidovudine and sulfamethoxazole-trimethoprim adversely affects the humoral immune response to Pneumocystis. PMID- 16467326 TI - Influence of exogenous reproductive hormones on specific antibody production in genital secretions after vaginal vaccination with recombinant cholera toxin B subunit in humans. AB - The objective of this study was to investigate the influence of exogenous reproductive hormones on the local and systemic production of specific immunoglobulin A (IgA) and IgG antibodies after vaginal vaccination with recombinant cholera toxin subunit B (CTB). Three groups of women using either progesterone-containing intrauterine devices (n=9), oral contraceptives (n=8), or no hormonal contraceptive methods (n=9) were vaginally immunized twice, 2 weeks apart. Cervical secretions, vaginal fluids, and serum were collected before and after vaccination. Total and CTB-specific IgA and IgG antibodies in genital secretions and serum were analyzed by enzyme-linked immunosorbent assay. A majority of the women presented strong CTB-specific IgA and IgG antibody responses in cervicovaginal secretions after vaccination, whereas the antitoxin responses in serum were weaker. Exogenously administered steroid hormones did not seem to have any impact on the production of specific antibodies. Both the frequencies and the magnitudes of IgA and IgG antitoxin responses in genital secretions were comparable among the three immunization groups. An association, in particular for IgA, was found between the magnitudes of the CTB-specific antibody responses in cervical secretions and vaginal fluids after vaccination. The sensitivities and positive predictive values of vaginal antibody analyses to reflect responses in cervical secretions were also high, suggesting that vaginal fluids alone might be used for evaluation of genital immune responses in large scale vaccination studies in the future. PMID- 16467327 TI - Neutralizing antibodies and persistence of immunity following anthrax vaccination. AB - Anthrax toxin consists of protective antigen (PA) and two toxic components, lethal factor (LF) and edema factor (EF). PA binds to mammalian cellular receptors and delivers the toxic components to the cytoplasm. PA is the primary antigenic component of the current anthrax vaccine. Immunity is due to the generation of antibodies that prevent the PA-mediated internalization of LF and EF. In this study, we characterized sera obtained from vaccinated military personnel. Anthrax vaccine is administered in a series of six injections at 0, 2, and 4 weeks and 6, 12, and 18 months, followed by annual boosters. The vaccination histories of the subjects were highly varied; many subjects had not completed the entire series, and several had not received annual boosters. We developed a simple colorimetric assay using alamarBlue dye to assess the antibody mediated neutralization of LF-mediated toxicity to the J774A.1 murine macrophage cell line. Recently vaccinated individuals had high antibody levels and neutralizing activity. One individual who had not been boosted for 5 years had low immunoglobulin G antibody levels but a detectable neutralization activity, suggesting that this individual produced low levels of very active antibodies. PMID- 16467328 TI - Performance of a time-resolved fluorescence immunoassay for measuring varicella zoster virus immunoglobulin G levels in adults and comparison with commercial enzyme immunoassays and Merck glycoprotein enzyme immunoassay. AB - Highly sensitive and specific, quantitative assays are needed to detect varicella zoster virus (VZV) immunoglobulin G in human sera, particularly for determining immune status and response following vaccination. A time-resolved fluorescence immunoassay (TRFIA) has been developed, and its performance was compared to that of two commercial enzyme immunoassays (EIAs) and Merck glycoprotein EIA (gpEIA). The TRFIA had equivalent sensitivity (97.8%) and high specificity (93.5%) in relation to gpEIA. A commercial (Behring) EIA compared favorably with TRFIA in terms of sensitivity (98.4%) but had lower specificity (80.7%). Another commercial EIA (Diamedix) had high specificity (97.1%) but low sensitivity (76.4%) compared to TRFIA if equivocal test results were treated as negative for VZV antibody. A novel feature of the TRFIA was that the cutoff was generated using population mixture modeling and was expressed in mIU/ml, as the assay was calibrated using the British standard VZV antibody. PMID- 16467329 TI - The probiotic bacterium Lactobacillus casei induces activation of the gut mucosal immune system through innate immunity. AB - The mechanisms by which probiotic bacteria affect the immune system are unknown yet, but many of them are attributed to an increase in the innate or in the acquired immune response. To study the influence of the probiotic bacterium Lactobacillus casei in the expression of receptors involved in the innate immune response, this bacterium was orally administered to BALB/c mice. After, they were sacrificed; the small intestine and intestinal fluids were collected to measure secretory immunoglobulin A (IgA) specific for L. casei. Mononuclear cells from Peyer's patches were isolated to determine the CD-206 and TLR-2 receptors. In histological slices we determined the number of IgA+, CD4+, CD8+, and CD3+ cells and two cytokines (interleulin-5 [IL-5] and IL-6). CD-206 and TLR-2 increased with respect to the untreated control. We did not observe an increase in the T population or in the IL-5-positive cells. IgA+ cells and IL-6-producing cells increased after 7 days of L. casei administration. We did not find specific antibodies against L. casei. The main immune cells activated after oral L. casei administration were those of the innate immune response, with an increase in the specific markers of these cells (CD-206 and TLR-2), with no modification in the number of T cells. PMID- 16467330 TI - Human interleukin-15 improves engraftment of human T cells in NOD-SCID mice. AB - Human nonobese diabetic-severe combined immune deficiency (NOD-SCID) mouse chimeras have been widely used as an in vivo model to assess human immune function. However, only a small fraction of transferred human T lymphocytes can be detected in human peripheral blood lymphocyte (huPBL)-NOD-SCID chimeras. To improve the reconstitution of human T lymphocytes in NOD-SCID mice, the use of recombinant human interleukin-15 (rhIL-15) as a stimulator of human lymphocytes was explored. Administration of rhIL-15 after transplantation of huPBLs into NOD SCID mice increased reconstitution of human T lymphocytes in a dose-dependent manner, with an optimal dosage of 1 microg/mouse. The number of human T lymphocytes (HLA-ABC+ CD3+) in the lymphoid organs or tissue of rhIL-15-treated huPBL-NOD-SCID mice increased 11- to 80-fold, and phytohemagglutinin-induced T lymphocyte proliferation and cytokine production were significantly enhanced. Additionally, although mature human cells have not been thought to enter the murine thymus, human T lymphocytes were detected in the huPBL-NOD-SCID thymus after rhIL-15 treatment. Thus, rhIL-15 can be used to optimize long-term peripheral T-cell engraftment in these human-mouse chimeras and may also be useful in clinical treatment of T-cell deficiencies. PMID- 16467331 TI - Performance of the TruGene human immunodeficiency virus type 1 genotyping kit and OpenGene DNA sequencing system on clinical samples diluted to approximately 100 copies per milliliter. AB - The TruGene human immunodeficiency virus type 1 (HIV-1) genotyping kit/OpenGene DNA sequencing system (Bayer HealthCare, Tarrytown, NY) reliably produced clinically acceptable resistance profiles for reverse transcriptase and protease inhibitors on patient samples diluted to approximately 100 copies/ml following extraction with the QIAamp viral RNA minikit (QIAGEN Inc., Valencia, CA). One modification of the standard protocol was made to guarantee PCR amplification: a centrifugation step to concentrate virus was added before RNA extraction. For genotypic antiretroviral resistance testing, no significant differences in the identification and sensitivity of detection for codon mutations, base mutations, and multibase sites were found between the original and diluted samples. PMID- 16467332 TI - Modulation of immunity-related gene expression in small intestines of mice by oral administration of lactoferrin. AB - Oral administration of lactoferrin (LF), an antimicrobial and immunomodulatory protein, shows a protective effect against infectious diseases, possibly via immunomodulation of the host. Initially, we confirmed an immunomodulatory effect of LF by observing changes in the number of cells in the leukocyte subsets in the peripheral blood and spleens of mice 1 day after oral administration of LF. Then we developed a quantitative reverse transcription-PCR method for 20 immunity related genes of antimicrobial proteins, pattern recognition receptors, cytokines, and lymphocyte mobilization-related proteins, and we assessed the expression of these genes in the small intestines of mice 2 h after administration of water, bovine serum albumin (BSA), or LF. Expression of the LF gene was lower in mice administered LF than in mice administered water or BSA, implying a negative-feedback control. Expression of gamma interferon (IFN-gamma) and interleukin-10 (IL-10) was lower in both BSA- and LF-administered mice than in water administered mice, suggesting a nonspecific effect of protein ingestion. Expression of NOD2, IFN-beta, and IL-12p40 was higher with LF administration than with water or BSA administration. The expression levels of these three genes were correlated. This study indicated that oral administration of LF modulates the small intestinal expression of genes closely related to the host defense in a specific or a nonspecific manner. PMID- 16467333 TI - Gamma interferon production in response to Mycobacterium bovis BCG and Mycobacterium tuberculosis antigens in infants born to human immunodeficiency virus-infected mothers. AB - In utero sensitization to infectious pathogens can establish immunological memory and may influence the immune response to unrelated antigens. Little is known about the influence of intrauterine human immunodeficiency virus (HIV) exposure on the cellular immune response to mycobacterial antigens. Whole-blood culture gamma interferon (IFN-gamma) production in response to mycobacterial antigens was measured at birth and 6 weeks of age to determine the characteristics of the IFN gamma response in HIV-exposed infants to Mycobacterium bovis BCG and mycobacterial antigens. At birth, we observed an increased immune activation in response to phytohemagglutinin among HIV-exposed, uninfected infants. In a proportion of these infants, we also observed an increased immune activation in response to purified protein derivative, BCG, and early secreted antigen target 6. Increases in the IFN-gamma response to the four antigens between birth and 6 weeks of age, observed in all HIV-unexposed infants, was absent in a substantial proportion of HIV-exposed, uninfected infants. The immunological differences persisted at 6 weeks of age, suggesting a sustained impact of in utero immune priming by HIV. Intrauterine exposure to HIV affects the infants' cellular immune response to mycobacterial antigens, either specifically or as a consequence of nonspecific, broadly reactive immune activation. Further studies will be important to help determine optimal vaccination and disease prevention strategies for this vulnerable population group. PMID- 16467334 TI - Cytokine response to antigen stimulation of whole blood from patients with Mycobacterium ulcerans disease compared to that from patients with tuberculosis. AB - Mycobacterium ulcerans disease (Buruli ulcer) is a skin-ulcerating infection common in some parts of the tropics. We have investigated cytokine secretion after stimulation of whole blood from Buruli ulcer (BU) patients in a region of endemicity in Ghana with M. ulcerans sonicate or culture filtrate antigens to investigate the development of the response over time and its specificity by comparison with the response to Mycobacterium tuberculosis sonicate in human immunodeficiency virus-negative tuberculosis patients. Significant gamma interferon (IFN-gamma) production in response to whole-blood stimulation with M. ulcerans sonicate was detected in patients with ulcers, which was higher than that in patients with nodules but similar to subjects with healed BU. The mean IFN-gamma response in household contacts of BU patients was not significantly different from that in healthy control subjects from an area of nonendemicity. Results in patients with untreated, smear-positive pulmonary tuberculosis and tuberculosis patients on treatment for more than 2 weeks showed that BU patients responded better to M. ulcerans antigens than tuberculosis patients. In contrast, interleukin-10 results were higher in patients with active M. ulcerans disease than in those with healed lesions, but the pattern of response was similar to that seen in tuberculosis. A similar pattern of cytokine secretion was found using M. tuberculosis sonicate as an antigen. Neither of the two culture filtrate antigens of M. ulcerans appeared to be more specific than M. ulcerans sonicate. In the early stages of M. ulcerans disease there was a mixed Th1 and Th2 cytokine response, but the Th1 response emerged as the dominant type. PMID- 16467335 TI - Role of CYP2E1 immunoglobulin G4 subclass antibodies and complement in pathogenesis of idiosyncratic drug-induced hepatitis. AB - Idiosyncratic drug-induced hepatitis (IDDIH) is the third most common cause for acute liver failure in the United States. Previous studies have attempted to identify susceptible patients or early stages of disease with various degrees of success. To determine if total serum immunoglobulin subclasses, CYP2E1-specific subclass autoantibodies, complement components, or immune complexes could distinguish persons with IDDIH from others exposed to drugs, we studied persons exposed to halogenated volatile anesthetics, which have been associated with IDDIH and CYP2E1 autoantibodies. We found that patients with anesthetic-induced IDDIH had significantly elevated levels of CYP2E1-specific immunoglobulin G4 (IgG4) autoantibodies, while anesthetic-exposed healthy persons had significantly elevated levels of CYP2E1-specific IgG1 autoantibodies. Anesthetic IDDIH patients had significantly lower levels of C4a, C3a, and C5a compared to anesthetic exposed healthy persons. C1q- and C3d-containing immune complexes were significantly elevated in anesthetic-exposed persons. In conclusion, our data suggest that anesthetic-exposed persons develop CYP2E1-specific IgG1 autoantibodies which may form detectable circulating immune complexes subsequently cleared by classical pathway activation of the complement system. Persons susceptible to anesthetic-induced IDDIH develop CYP2E1-specific IgG4 autoantibodies which form small, nonprecipitating immune complexes that escape clearance because of their size or by direct inhibition of complement activation. PMID- 16467336 TI - Evaluation of a tetraplex microsphere assay for Bordetella pertussis antibodies. AB - To increase testing efficiency, a microsphere-based multianalyte immune detection (MAID) system was developed to measure serum immunoglobulin G (IgG) and IgA recognizing two Bordetella pertussis antigens, pertussis toxin (PT) and filamentous hemagglutinin antigen (FHA). The assay was performed as two separate duplexes. One duplex measured IgG to PT and FHA, and the other measured IgA to PT and FHA. The two duplexes were then combined and analyzed as a tetraplex. The MAID system and an in-house enzyme-linked immunosorbent assay (ELISA) system were used to evaluate 100 sera from blood donors and 220 consecutive sera submitted for B. pertussis antibody testing. For both the MAID and ELISA systems, antibody levels were defined as increased if greater than the blood donor group 95th percentile value. The qualitative concordance rates between MAID and ELISA results for the 220 consecutively submitted sera were as follows: PT IgG, 99%; PT IgA, 94%; FHA IgG, 93%; FHA IgA, 94%. The overall concordance rate was 95% (836 of 880 result sets). For 29 of 44 (66%) discordant result sets, the discordant MAID result was supported by the MAID and ELISA results for other B. pertussis antibodies. The MAID and in-house ELISA systems were also used to evaluate 20 sera previously tested for pertussis antibodies at a pertussis vaccine research laboratory; MAID results for all four analytes did not significantly differ from results obtained by the research laboratory. These findings show that antibodies to B. pertussis antigens can be measured easily and accurately using a tetraplex microsphere system. PMID- 16467337 TI - Kinetics and diagnostic and prognostic potential of quantitative Western blot analysis and antigen-specific enzyme-linked immunosorbent assay in experimental canine leishmaniasis. AB - Quantitative computerized Western blot analysis of antibody responses during experimental canine Leishmania infantum infection distinguished between immunodominant and nonimmunodominant protein bands. Six infected beagles, positive by both PCR and parasite culture, were monitored over 75 weeks postinfection and during a 12-week allopurinol treatment course. All dogs were symptomatic at the time of treatment. Of 12 antigenic bands examined, the immunodominant bands (12, 14, 24, 29, 48, and 68 kDa) showed significantly increased intensities (P<0.01) and higher frequencies of recognition than the nonimmunodominant bands at all time points. Detection of the former bands at 6 weeks postinfection preceded seroconversion by enzyme-linked immunosorbent assay (ELISA) both on crude Leishmania antigen or the recombinant proteins rK39 and HSP70. Reactivity with the 14-, 48-, and 68-kDa bands signified early infection, whereas increased reactivity with the 14-, 24-, and 29-kDa bands was associated with posttreatment parasite persistence and potential unfavorable prognosis. Total lane intensity (TLI) emerged as a sensitive marker for early infection and increased as early as 4 weeks postinfection. TLI had a significantly higher (P<0.01) relative increase rate than crude Leishmania antigen or HSP70 or rK39 ELISA at all time points. These immunodominant antigens and TLI, as determined by quantitative Western blotting, will be valuable for early detection and treatment evaluation of canine leishmaniasis. PMID- 16467338 TI - Detection of kappa and lambda light chain monoclonal proteins in human serum: automated immunoassay versus immunofixation electrophoresis. AB - Recently, turbidimetric immunoassays for detecting and quantifying kappa and lambda free light chains (FLC) have become available and are promoted as being more sensitive than immunofixation electrophoresis (IFE) in detecting FLC monoclonal proteins. In this study, we assessed the ability of these turbidimetric assays to detect serum monoclonal proteins involving both free and heavy-chain-bound kappa and lambda light chains compared to standard immunofixation electrophoresis. Sera demonstrating a restricted band of protein migration (other than a definite M spike) by serum protein electrophoresis (SPE), which may represent early monoclonal proteins, were also examined. When compared to IFE, percent agreement, sensitivity, and specificity for the kappa-FLC and lambda-FLC were 94.6, 72.9, and 99.5% and 98.5, 91.4, and 99.7%, respectively, in detecting monoclonal proteins involving free and heavy-chain-bound light chains. The majority of sera (73.7%) demonstrating a restricted band of protein migration on SPE demonstrated abnormal IFE patterns suggestive of multiple myeloma or monoclonal gammopathy of unknown significance, but gave normal kappa/lambda FLC ratios using the turbidimetric immunoassays. In conclusion, the kappa and lambda FLC assays are significantly less sensitive (72.9 to 91.4%) than IFE, but specific in detecting serum monoclonal proteins. Moreover, the kappa/lambda ratio has little value in routine screening since the majority of sera with abnormal IFE patterns had normal kappa/lambda FLC ratios. PMID- 16467339 TI - General anesthesia delays the inflammatory response and increases survival for mice with endotoxic shock. AB - Anesthesia is an indispensable component of any operative procedure. In this study, we demonstrate that continuous isoflurane anesthesia for 1 h after a lethal dose (20 mg/kg of body weight) of Escherichia coli lipopolysaccharide (LPS) results in a significant increase in survival of C57BL/6J (B6) mice in comparison with survival of nonanesthetized mice. Protection by anesthesia correlates with a delay in plasma LPS circulation, resulting in a delayed inflammatory response, particularly DNA binding activity of NF-kappaB and serum levels of tumor necrosis factor alpha, interleukin-6 (IL-6), and IL-10. Disparate classes of anesthetic agents produce the same effects on the inflammatory response, which is also independent of the inbred mouse strain used. These results suggest that anesthesia has an important impact on the outcome from endotoxemia. Moreover, the immunomodulatory effects of anesthetics should be considered when interpreting data from experimental animal models. PMID- 16467340 TI - Anti-interleukin-15 prevents arthritis in Borrelia-vaccinated and -infected mice. AB - We showed previously that interleukin-17 (IL-17) plays a significant role in the induction of arthritis associated with Borrelia vaccination and challenge. Little information, however, is available about the chain of immunologic events that leads to the release of IL-17. The production of IL-17 has been linked to stimulation of memory cells by IL-15. Therefore, we hypothesized that IL-15 is involved in the induction of arthritis associated with Borrelia vaccination and infection of mice. Here we present evidence that treatment of Borrelia-vaccinated and -infected mice with anti-IL-15 antibody prevents swelling of the hind paws. More importantly, both anti-IL-15 antibody- and recombinant IL-15 receptor alpha treated Borrelia-vaccinated and -infected mice were free of major histopathologic indications of arthritis, including hyperplasia, hypertrophy, and vilus formation of the synovium. Similarly, the synovial space and perisynovium were free of inflammatory cells. By contrast, the synovium of nontreated Borrelia-vaccinated and -infected mice had overt hyperplasia, hypertrophy, and vilus formation. Moreover, the synovial space and perisynovium were infiltrated with neutrophils, macrophages, and lymphocytes. Finally, we show that recombinant IL-15 stimulates the release of IL-17 from lymph node cells obtained near the arthritic site. These results suggest that IL-15 plays a major role in orchestrating IL-17 induction of arthritis associated with Borrelia-vaccinated and -infected mice. PMID- 16467341 TI - Development of specific immunoglobulin Ga (IgGa) and IgGb antibodies correlates with control of parasitemia in Babesia equi Infection. AB - In this study, the kinetics of specific immunoglobulin G (IgG) isotypes were characterized in Babesia equi (Theileria equi)-infected horses. IgGa and IgGb developed during acute infection, whereas IgG(T) was detected only after resolution of acute parasitemia. The same IgG isotype profile induced during acute infection was obtained by equi merozoite antigen 1/saponin immunization. PMID- 16467343 TI - Different positive predictive values of commercially available human immunodeficiency virus enzyme-linked immunosorbent assays. PMID- 16467342 TI - Evaluation of sedimentation rate, rheumatoid factor, C-reactive protein, and tumor necrosis factor for the diagnosis of infective endocarditis. PMID- 16467344 TI - Report refuting value of immune complexes to diagnose Lyme disease is invalid. PMID- 16467345 TI - Determinants of primary medical care quality measured under the new UK contract: cross sectional study. AB - OBJECTIVE: To identify factors associated with the quality of primary medical care incentivised under the new UK general medical services contract. DESIGN: Cross sectional study. SETTING: NHS Ayrshire and Arran area, Scotland. PARTICIPANTS: 60 general practices. MAIN OUTCOME MEASURES: Quality scores reflecting the total points achieved on the 10 clinical domains and holistic care. Univariate and multivariate regression analyses were used to relate quality scores to measures of population characteristics, urban-rural location, general practitioner characteristics, clinical team size and composition, practice characteristics, and income from other sources. RESULTS: Deprivation was associated with higher scores. Quality scores increased with the size of the clinical team. Practices with higher income from other sources had lower quality scores. Practices that were accredited, had training status, or contained younger general practitioners had higher quality scores, but these effects were explained by other associated factors. 53% of the variation in quality scores was explained by a multivariate model, which included measures of deprivation, clinical team size and composition, and financial incentives. CONCLUSIONS: Population characteristics showed little association with the quality of primary medical care incentivised under the UK general medical services contract. Larger clinical teams delivered higher quality clinical care, but the nurse-doctor composition of the clinical team did not influence quality. Practices that were more likely to respond to financial incentives because of previous behaviour or lower income from other sources recorded higher quality. If generalisable, the results suggest that initiatives to improve primary medical care quality should focus on the structure and resourcing of providers. PMID- 16467346 TI - Factors influencing death at home in terminally ill patients with cancer: systematic review. AB - OBJECTIVES: To determine the relative influence of different factors on place of death in patients with cancer. DATA SOURCES: Four electronic databases-Medline (1966-2004), PsycINFO (1972-2004), CINAHL (1982-2004), and ASSIA (1987-2004); previous contacts with key experts; hand search of six relevant journals. REVIEW METHODS: We generated a conceptual model, against which studies were analysed. Included studies had original data on risk factors for place of death among patients, > 80% of whom had cancer. Strength of evidence was assigned according to the quantity and quality of studies and consistency of findings. Odds ratios for home death were plotted for factors with high strength evidence. RESULTS: 58 studies were included, with over 1.5 million patients from 13 countries. There was high strength evidence for the effect of 17 factors on place of death, of which six were strongly associated with home death: patients' low functional status (odds ratios range 2.29-11.1), their preferences (2.19-8.38), home care (1.37-5.1) and its intensity (1.06-8.65), living with relatives (1.78-7.85), and extended family support (2.28-5.47). The risk factors covered all groups of the model: related to illness, the individual, and the environment (healthcare input and social support), the latter found to be the most important. CONCLUSIONS: The ne of factors that influence where patients with cancer die is complicated. Future policies and clinical practice should focus on ways of empowering families and public education, as well as intensifying home care, risk assessment, and training practitioners in end of life care. PMID- 16467347 TI - Improving tuberculosis control through public-private collaboration in India: literature review. AB - OBJECTIVE: To review the characteristics of public-private mix projects in India and their effect on case notification and treatment outcomes for tuberculosis. DESIGN: Literature review. DATA SOURCES: Review of surveillance records from Indian tuberculosis programme project, evaluation reports, and medical literature for public-private mix projects in India. DATA EXTRACTION: Project characteristics, tuberculosis case notification of new patients with sputum smear results positive for acid fast bacilli, and treatment outcome. DATA SYNTHESIS: Of 24 identified public-private mix projects, data were available from 14 (58%), involving private practitioners, corporations, and non-governmental organisations. In all reviewed projects, the public sector tuberculosis programme provided training and supervision of private providers. Among the five projects with available data on historical controls, case notification rates were higher after implementation of a public-private mix project. Among seven projects involving private practitioners, 2796 of 12 147 (23%) new patients positive for acid fast bacilli were attributed to private providers. Corporate based and non governmental organisations served as the main source for tuberculosis programme services in seven project areas, detecting 9967 new patients positive for acid fast bacilli. In nine of 12 projects with data on treatment outcomes, private providers exceeded the programme target of 85% treatment success for new patients positive for acid fast bacilli. CONCLUSIONS: Public-private mix activities were associated with increased case notification, while maintaining acceptable treatment outcomes. Collaborations between public and private providers of health care hold considerable potential to improve tuberculosis control in India. PMID- 16467348 TI - Oral protein energy supplements for children with cystic fibrosis: CALICO multicentre randomised controlled trial. AB - OBJECTIVE: To determine whether oral protein energy supplements, used long term in children with cystic fibrosis who are moderately malnourished, improve nutritional and other outcomes. DESIGN: Multicentre randomised controlled trial. SETTING: Seven specialist paediatric cystic fibrosis centres and their associated shared care clinics and seven smaller paediatric cystic fibrosis clinics. PARTICIPANTS: 102 children with cystic fibrosis, aged between 2 and 15 years, who were moderately malnourished. INTERVENTIONS: Oral protein energy supplements in addition to usual dietary advice compared with dietary advice alone, for 12 months. MAIN OUTCOME MEASURE: Change in body mass index centile over one year. RESULTS: Use of supplements was not associated with a change in body mass index centile (mean difference 2.99 centile points, 95% confidence interval -2.70 to 8.68) or other nutritional and spirometric outcomes in this group of children. CONCLUSIONS: Long term use of oral protein energy supplements did not result in an improvement in nutritional status or other clinical outcomes in children with cystic fibrosis who were moderately malnourished. Oral protein energy supplements should not be regarded as an essential part of the management of this group of children. TRIAL REGISTRATION: ISRCTN: 95744468. PMID- 16467349 TI - Regional and cellular gene expression changes in human Huntington's disease brain. AB - Huntington's disease (HD) pathology is well understood at a histological level but a comprehensive molecular analysis of the effect of the disease in the human brain has not previously been available. To elucidate the molecular phenotype of HD on a genome-wide scale, we compared mRNA profiles from 44 human HD brains with those from 36 unaffected controls using microarray analysis. Four brain regions were analyzed: caudate nucleus, cerebellum, prefrontal association cortex [Brodmann's area 9 (BA9)] and motor cortex [Brodmann's area 4 (BA4)]. The greatest number and magnitude of differentially expressed mRNAs were detected in the caudate nucleus, followed by motor cortex, then cerebellum. Thus, the molecular phenotype of HD generally parallels established neuropathology. Surprisingly, no mRNA changes were detected in prefrontal association cortex, thereby revealing subtleties of pathology not previously disclosed by histological methods. To establish that the observed changes were not simply the result of cell loss, we examined mRNA levels in laser-capture microdissected neurons from Grade 1 HD caudate compared to control. These analyses confirmed changes in expression seen in tissue homogenates; we thus conclude that mRNA changes are not attributable to cell loss alone. These data from bona fide HD brains comprise an important reference for hypotheses related to HD and other neurodegenerative diseases. PMID- 16467350 TI - The mitochondrial ATP-binding cassette transporter Abcb7 is essential in mice and participates in cytosolic iron-sulfur cluster biogenesis. AB - Proteins with iron-sulfur (Fe-S) clusters participate in multiple metabolic pathways throughout the cell. The mitochondrial ABC half-transporter Abcb7, which is mutated in X-linked sideroblastic anemia with ataxia in humans, is a functional ortholog of yeast Atm1p and is predicted to export a mitochondrially derived metabolite required for cytosolic Fe-S cluster assembly. Using an inducible Cre/loxP system to delete exons 9 and 10 of the Abcb7 gene, we examined the phenotype of mice deficient in Abcb7. We found that Abcb7 was essential in extra-embryonic tissues early in gestation and that the mutant allele exhibits an X-linked parent-of-origin lethality effect. Furthermore, using X-chromosome inactivation assays and tissue-specific deletions, Abcb7 was found to be essential for the development and function of numerous other cell types and tissues. A notable exception to this was liver, where loss of Abcb7 impaired cytosolic Fe-S cluster assembly but was not lethal. In this situation, control of iron regulatory protein 1, a key cytosolic modulator of iron metabolism, which is responsive to the availability of cytosolic Fe-S clusters, was impaired and contributed to the dysregulation of hepatocyte iron metabolism. Altogether, these studies demonstrate the essential nature of Abcb7 in mammals and further substantiate a central role for mitochondria in the biogenesis of cytosolic Fe-S proteins. PMID- 16467351 TI - Opposite effects of daylength and temperature on flowering and summer dormancy of Poa bulbosa. AB - BACKGROUND AND AIMS: The timing of flowering and summer dormancy induction plays a central role in the adaptation of Mediterranean geophytes to changes in the length of the growth season along rainfall gradients. Our aim was to analyse the role of the variation in the responses of flowering and summer dormancy to vernalization, daylength and growth temperature for the adaptation of Poa bulbosa, a perennial geophytic grass, to increasing aridity. METHODS: Flowering and dormancy were studied under controlled daylengths [9 h short day (SD) vs. 16 h long day (LD)] and temperatures (16/10, 22/16 and 28/22 degrees C day/night) in four ecotypes originating in arid, semi-arid and mesic habitats (110, 276 and 810 mm rain year(-1), respectively) and differing in flowering capacity under natural conditions: arid-flowering, semi-arid-flowering, semi-arid-non-flowering and mesic-non-flowering. KEY RESULTS: Flowering and dormancy were affected in opposite ways by daylength and growth temperature. Flowering occurred almost exclusively under SD. In contrast, plants became dormant much earlier under LD than under SD. In both daylengths, high temperature and pre-chilling (6 weeks at 5 degrees C) enhanced dormancy imposition, but inhibited or postponed flowering, respectively. Induction of flowering and dormancy in the different ecotypes showed differential responsiveness to daylength and temperature. Arid and semi arid ecotypes had a higher proportion of flowering plants and flowering tillers as well as more panicles per plant than mesic ecotypes. 'Flowering' ecotypes entered dormancy earlier than 'non-flowering' ecotypes, while the more arid the site of ecotype origin, the earlier the ecotype entered dormancy. CONCLUSIONS: Variation in the flowering capacity of ecotypes differing in drought tolerance was interpreted as the result of balanced opposite effects of daylength and temperature on the flowering and dormancy processes. PMID- 16467353 TI - Structural aspects and ecophysiology of anther opening in Allium triquetrum. AB - BACKGROUND AND AIMS: Tissue desiccation is considered to be involved in anther opening, and it is agreed that environmental humidity affects its timing. Different sources of evidence suggest that the later steps of the process (i.e. stomium opening and outward wall bending) are regulated in different ways. Anther opening was studied in Allium triquetrum under four regimes of relative humidity (RH) to analyse the effect of this parameter and to speculate about its possible regulation. METHODS: Anther histology was studied in cross-sections under a microscope. The times of visible anther opening and complete outward wall bending were recorded separately for each level of RH. Frequency distributions were plotted to express anther behaviour. KEY RESULTS: When a longitudinal stomium breaks the anther remains closed due to adherence of walls on each side of the stomium. Anther opening occurs when the adhering walls subsequently separate. Later, the walls shrink laterally and bend outward. The anthers of the inner whorl opened during the morning of the first day of anthesis, while those of the outer whorl opened during the afternoon. Low RH (20 %) did not cause any evident acceleration of anther opening, but it did cause delay and inhibition of the opening of some anthers in the outer whorl. High RH (55 and 98 %) caused different degrees of delay and also inhibition of anther opening, but most anthers opened within the expected range of time. The time taken for outward wall bending was shortened at 20 % RH. Anther wall outward bending was inhibited at 55 % and 98 % RH. CONCLUSIONS: Anther opening occurred at a specific moment of anther development, separated in time from stomium breakage, and seemed related to dehydration caused by reabsorption of water by contiguous tissues. Outward bending of the wall was facilitated by evaporation. Anther opening and anther wall outward bending seemed to be regulated differently in relation to water control. PMID- 16467354 TI - B-type natriuretic peptide in paediatric patients with congenital heart disease. AB - AIMS: To examine the diagnostic value of B-type natriuretic peptide (BNP) plasma concentration in congenital heart disease. METHODS AND RESULTS: BNP was measured in 288 consecutive patients (mean age 6.0 +/- 6.4 years) with left-to-right shunt, left or right heart obstruction, tetralogy of Fallot, functionally univentricular heart, or impaired left ventricular function and compared with age and gender-specific normal values, and to haemodynamic and echocardiographic data. BNP increased with decreasing left ventricular shortening fraction (r = 0.80; P < 0.001). In patients with left-to-right shunt, BNP was increased (mean SDS +1.64; P < 0.001) and positively correlated (P < 0.001) to shunt volume (r = 0.66), systolic right ventricular pressure (r = 0.69), mean pressure of the pulmonary artery (r = 0.66), and pulmonary resistance (r = 0.59). There was no correlation between BNP and invasive pressure gradient or extent of ventricular hypertrophy in patients with left or right heart obstruction. In patients with tetralogy of Fallot, BNP was not significantly increased. Patients with functionally univentricular heart had elevated BNP plasma levels (mean SDS +1.39; P < 0.001) without decrease after volume unloading by cavopulmonary connection. CONCLUSION: In children with congenital heart defects, plasma BNP correlates closely to ventricular function. BNP plasma levels do not reflect directly the extent of ventricular pressure or volume work, but mirror the impairment of the loaded ventricles. Normal BNP cannot exclude pathology, but reflects a compensated status of the heart. PMID- 16467352 TI - Cytoskeleton and morphogenesis in brown algae. AB - BACKGROUND: Morphogenesis on a cellular level includes processes in which cytoskeleton and cell wall expansion are strongly involved. In brown algal zygotes, microtubules (MTs) and actin filaments (AFs) participate in polarity axis fixation, cell division and tip growth. Brown algal vegetative cells lack a cortical MT cytoskeleton, and are characterized by centriole-bearing centrosomes, which function as microtubule organizing centres. SCOPE: Extensive electron microscope and immunofluorescence studies of MT organization in different types of brown algal cells have shown that MTs constitute a major cytoskeletal component, indispensable for cell morphogenesis. Apart from participating in mitosis and cytokinesis, they are also involved in the expression and maintenance of polarity of particular cell types. Disruption of MTs after Nocodazole treatment inhibits cell growth, causing bulging and/or bending of apical cells, thickening of the tip cell wall, and affecting the nuclear positioning. Staining of F-actin using Rhodamine-Phalloidin, revealed a rich network consisting of perinuclear, endoplasmic and cortical AFs. AFs participate in mitosis by the organization of an F-actin spindle and in cytokinesis by an F-actin disc. They are also involved in the maintenance of polarity of apical cells, as well as in lateral branch initiation. The cortical system of AFs was found related to the orientation of cellulose microfibrils (MFs), and therefore to cell wall morphogenesis. This is expressed by the coincidence in the orientation between cortical AFs and the depositing MFs. Treatment with cytochalasin B inhibits mitosis and cytokinesis, as well as tip growth of apical cells, and causes abnormal deposition of MFs. CONCLUSIONS: Both the cytoskeletal elements studied so far, i.e. MTs and AFs are implicated in brown algal cell morphogenesis, expressed in their relationship with cell wall morphogenesis, polarization, spindle organization and cytokinetic mechanism. The novelty is the role of AFs and their possible co-operation with MTs. PMID- 16467355 TI - Personal services. PMID- 16467356 TI - The role of vasopressin in cardiorespiratory arrest and pulmonary hypertension. AB - Vasopressin is a peptide synthesized in the hypothalamus whose primary role is in fluid homeostasis. It has recently gained interest as a potential agent in the treatment of cardiorespiratory arrest. Initial human studies have shown benefits with vasopressin in patients with out of hospital ventricular fibrillation and asystolic cardiac arrest. One subgroup of patients not included in these trials is patients with pulmonary hypertension, who have a five-year mortality rate of 50%. Animal studies have shown vasopressin to be a vasodilator in the pulmonary vascular system of rats, under normoxic and hypoxic conditions, with conflicting results in canines. Human studies have shown conflicting results with increases, decreases and no changes seen in pulmonary artery pressures of patients with a variety of clinical conditions. Research needs to be done in patients with pulmonary hypertension regarding the potential role of vasopressin during cardiac arrest in this subgroup. PMID- 16467357 TI - Determination of cell fate along the anteroposterior axis of the Drosophila ventral midline. AB - The Drosophila ventral midline has proven to be a useful model for understanding the function of central organizers during neurogenesis. The midline is similar to the vertebrate floor plate, in that it plays an essential role in cell fate determination in the lateral CNS and also, later, in axon pathfinding. Despite the importance of the midline, the specification of midline cell fates is still not well understood. Here, we show that most midline cells are determined not at the precursor cell stage, but as daughter cells. After the precursors divide, a combination of repression by Wingless and activation by Hedgehog induces expression of the proneural gene lethal of scute in the most anterior midline daughter cells of the neighbouring posterior segment. Hedgehog and Lethal of scute activate Engrailed in these anterior cells. Engrailed-positive midline cells develop into ventral unpaired median (VUM) neurons and the median neuroblast (MNB). Engrailed-negative midline cells develop into unpaired median interneurons (UMI), MP1 interneurons and midline glia. PMID- 16467358 TI - Hand, an evolutionarily conserved bHLH transcription factor required for Drosophila cardiogenesis and hematopoiesis. AB - The Hand gene family encodes highly conserved basic helix-loop-helix (bHLH) transcription factors that play crucial roles in cardiac and vascular development in vertebrates. In Drosophila, a single Hand gene is expressed in the three major cell types that comprise the circulatory system: cardioblasts, pericardial nephrocytes and lymph gland hematopoietic progenitors, but its function has not been determined. Here we show that Drosophila Hand functions as a potent transcriptional activator, and converting it into a repressor blocks heart and lymph gland formation. Disruption of Hand function by homologous recombination also results in profound cardiac defects that include hypoplastic myocardium and a deficiency of pericardial and lymph gland hematopoietic cells, accompanied by cardiac apoptosis. Targeted expression of Hand in the heart completely rescued the lethality of Hand mutants, and cardiac expression of a human HAND gene, or the caspase inhibitor P35, partially rescued the cardiac and lymph gland phenotypes. These findings demonstrate evolutionarily conserved functions of HAND transcription factors in Drosophila and mammalian cardiogenesis, and reveal a previously unrecognized requirement of Hand genes in hematopoiesis. PMID- 16467359 TI - Sfrp1 and Sfrp2 regulate anteroposterior axis elongation and somite segmentation during mouse embryogenesis. AB - Regulation of Wnt signaling is essential for embryonic patterning. Sfrps are secreted Wnt antagonists that directly interact with the Wnt ligand to inhibit signaling. Here, we show that Sfrp1 and Sfrp2 are required for anteroposterior (AP) axis elongation and somitogenesis in the thoracic region during mouse embryogenesis. Double homozygous mutations in Sfrp1 and Sfrp2 lead to severe shortening of the thoracic region. By contrast, a homozygous mutation in one or the other exerts no effect on embryogenesis, indicating that Sfrp1 and Sfrp2 are functionally redundant. The defect of a shortened thoracic region appears to be the consequence of AP axis reduction and incomplete somite segmentation. The reduction in the AP axis is partially due to abnormalities in cell migration of pre-somitic mesoderm from the end of gastrulation. Aberrant somite segmentation is associated with altered oscillations of Notch signaling, as evidenced by abnormal Lfng and Hes7 expression during somitogenesis in the thoracic region. This study suggests that Wnt regulation by Sfrp1 and Sfrp2 is required for embryonic patterning. PMID- 16467360 TI - C/EBPalpha is required for lung maturation at birth. AB - Epithelial cells lining the peripheral lung synthesize pulmonary surfactant that reduces surface tension at the air-liquid interface. Lack of surfactant lipids and proteins in the lungs causes respiratory distress syndrome, a common cause of morbidity and mortality in preterm infants. We show that C/EBPalpha plays a crucial role in the maturation of the respiratory epithelium in late gestation, being required for the production of surfactant lipids and proteins necessary for lung function. Deletion of the Cebpa gene in respiratory epithelial cells in fetal mice caused respiratory failure at birth. Structural and biochemical maturation of the lung was delayed. Normal synthesis of surfactant lipids and proteins, including SP-A, SP-B, SP-C, SP-D, ABCA3 (a lamellar body associated protein) and FAS (precursor of fatty acid synthesis) were dependent upon expression of the C/EBPalpha in respiratory epithelial cells. Deletion of the Cebpa gene caused increased expression of Tgfb2, a growth factor that inhibits lung epithelial cell proliferation and differentiation. Normal expression of C/EBPalpha required Titf1 and Foxa2, transcription factors that also play an important role in perinatal lung differentiation. C/EBPalpha participates in a transcriptional network that is required for the regulation of genes mediating perinatal lung maturation and surfactant homeostasis that is necessary for adaptation to air breathing at birth. PMID- 16467361 TI - Semaphorin 3d guides laterality of retinal ganglion cell projections in zebrafish. AB - The optic chiasm is an important choice point at which retinal ganglion cell (RGC) axons either cross the midline to innervate the contralateral brain or turn back to innervate the ipsilateral brain. Guidance cues that regulate this decision, particularly those directing the midline crossing of contralateral axons, are still not well understood. Here we show that Sema3d, a secreted semaphorin expressed at the midline, guides the crossing of RGC axons in zebrafish. Both Sema3d knockdown and ubiquitous overexpression induced aberrant ipsilateral projections, suggesting that Sema3d normally guides axons into the contralateral optic tract. Live imaging in vivo showed that RGC growth cones responded to ubiquitous Sema3d overexpression by pausing for extended periods and increasing their exploratory behavior at the midline, suggesting that Sema3d overexpression causes the midline environment to become less favorable for RGC axon extension. Interestingly, Sema3d overexpression did not affect growth cone behaviors before the midline, suggesting that RGC axons normally respond to Sema3d only upon reaching the midline. After Sema3d knockdown, growth cones grew across the midline but then paused or repeatedly retracted, impairing their ability to leave the midline region. Our results indicate that a proper balance of Sema3d is needed at the midline for the progression of RGC axons from the chiasm midline into the contralateral optic tract. PMID- 16467362 TI - Somite-derived cells replace ventral aortic hemangioblasts and provide aortic smooth muscle cells of the trunk. AB - We have previously shown that endothelial cells of the aortic floor give rise to hematopoietic cells, revealing the existence of an aortic hemangioblast. It has been proposed that the restriction of hematopoiesis to the aortic floor is based on the existence of two different and complementary endothelial lineages that form the vessel: one originating from the somite would contribute to the roof and sides, another from the splanchnopleura would contribute to the floor. Using quail/chick orthotopic transplantations of paraxial mesoderm, we have traced the distribution of somite-derived endothelial cells during aortic hematopoiesis. We show that the aortic endothelium undergoes two successive waves of remodeling by somitic cells: one when the aortae are still paired, during which the initial roof and sides of the vessels are renewed; and a second, associated to aortic hematopoiesis, in which the hemogenic floor is replaced by somite endothelial cells. This floor thus appears as a temporary structure, spent out and replaced. In addition, the somite contributes to smooth muscle cells of the aorta. In vivo lineage tracing experiments with non-replicative retroviral vectors showed that endothelial cells do not give rise to smooth muscle cells. However, in vitro, purified endothelial cells acquire smooth muscle cells characteristics. Taken together, these data point to the crucial role of the somite in shaping the aorta and also give an explanation for the short life of aortic hematopoiesis. PMID- 16467364 TI - The effect of UVB on lupus skin: new light on the role of apoptosis in the pathogenesis of autoimmunity. PMID- 16467366 TI - An international consensus survey of the diagnostic criteria for juvenile dermatomyositis (JDM). AB - OBJECTIVE: To develop revised criteria for the diagnosis of juvenile dermatomyositis (JDM) using an international consensus process. METHODS: An initial survey was circulated to members of the Network for JDM and the Paediatric Rheumatology International Trials Organisation (PRINTO). Each individual was asked to identify those criteria that were felt to be most helpful in the diagnosis of classical JDM. A second survey was derived from these results and used to rank these proposed criteria in order of their importance and usefulness in clinical practice. RESULTS: The first survey had a response rate of 49.8% (118 individuals) from 92 centres in 32 countries. All responders routinely used proximal muscle weakness and characteristic skin rash in the diagnosis of JDM, while 86.8% used elevated muscle enzymes. Muscle biopsy, magnetic resonance imaging (MRI) and changes on the electromyogram (EMG) were deemed important diagnostic criteria. Other criteria, including myositis-specific or -related antibodies, nailfold capillaroscopy, factor VIII-related antigen, muscle ultrasound, calcinosis and neopterin, were used by 35.3% of respondents. Seventy eight respondents to the first survey (66%) responded to the second survey. Typical MRI and muscle biopsy changes were rated by all to be the most useful clinically relevant diagnostic criteria after proximal muscle weakness, characteristic skin rash and elevated muscle enzymes. These were followed by myopathic changes on EMG, calcinosis, dysphonia and nailfold capillaroscopy, which were ranked equally. CONCLUSION: This process identified nine criteria that clinicians felt to be helpful or important in the diagnosis of JDM. A further process of refinement and validation is necessary to agree an internationally acceptable, clinically usable set of diagnostic criteria. PMID- 16467367 TI - Retinoic acid and oncostatin M combine to promote cartilage degradation via matrix metalloproteinase-13 expression in bovine but not human chondrocytes. AB - OBJECTIVES: Retinoic acid (RetA) and oncostatin M (OSM) have both been shown to mediate potent effects with respect to extracellular matrix integrity. This study assesses the effects of a RetA + OSM combination on cartilage catabolism. METHODS: Animal and human cartilage samples were used to assess the ability of RetA + OSM to promote the release of collagen and proteoglycan fragments, which was determined by measuring glycosaminoglycan and hydroxyproline, respectively. Total collagenolytic and tissue inhibitor of metalloproteinases (TIMP) inhibitory activities were determined by bioassay, whilst gene expression of matrix metalloproteinases (MMPs) and TIMP-1 were determined by northern blotting. Immunohistochemistry was used to assess the presence of MMP-1 and -13 in resorbing cartilage explants. RESULTS: Both agents alone induced proteoglycan release from bovine cartilage, whilst RetA-induced collagen release was variable. Reproducible and synergistic collagenolysis was observed with RetA + OSM, which appeared to be due to MMP-13. Similar collagen release was observed from porcine cartilage. Conversely, no collagen release was seen with human articular cartilage. In primary human chondrocytes, RetA + OSM failed to induce MMP-1 or 13 but caused a significant increase in TIMP-1 expression. CONCLUSIONS: These novel observations show that the combination of RetA + OSM has profound effects on cartilage matrix turnover, but these effects are species-specific. A better understanding of the mechanism by which this combination differentially regulates MMP and TIMP expression in human chondrocytes could provide valuable insight into new therapeutic strategies aimed at the prevention of cartilage destruction. PMID- 16467368 TI - Comparative study of the antimicrobial activity of bis(Nalpha-caproyl-L-arginine) 1,3-propanediamine dihydrochloride and chlorhexidine dihydrochloride against Staphylococcus aureus and Escherichia coli. AB - OBJECTIVES: The aim of this study is to gain insight into the mechanism of the antimicrobial action of a novel arginine-based surfactant, bis(N(alpha)-caproyl-L arginine)-1,3-propanediamine dihydrochloride [C(3)(CA)(2)]. METHODS: To this end, we compared its effects against Staphylococcus aureus and Escherichia coli with those caused by the commercial and widely known antiseptic, chlorhexidine dihydrochloride (CHX). RESULTS: Both disrupted the cell membrane of the target bacteria to cause potassium leakage and morphological damage. The effect of C(3)(CA)(2) on E. coli was concentration dependent, causing loss of membrane potential and membrane integrity leading to cell death, whereas CHX did not have these effects on E. coli. The effect of C(3)(CA)(2) on S. aureus was the formation of mesomes and cytoplasmic clear zones, but the loss of membrane potential and membrane integrity was slightly lower than that with CHX. CONCLUSIONS: We propose that C(3)(CA)(2) acts preferentially against Gram negative bacteria through strong initial binding to the surface lipopolysaccharides and subsequently partitioning into the cell membrane to cause membrane damage, followed by cell death. PMID- 16467370 TI - Chronic stress accelerates learning and memory impairments and increases amyloid deposition in APPV717I-CT100 transgenic mice, an Alzheimer's disease model. AB - Although chronic stress is known to be linked with memory and other neurological disorders, little is known about the relationship between chronic stress and the onset or development of Alzheimer's disease (AD). In this study, we investigated the effects of long-term stress on the onset and severity of cognitive deficits and pathological changes in APPV717I-CT100 mice overexpressing human APP-CT100 containing the London mutation (V717I) after exposure to immobilization stress. We found that chronic immobilization stress accelerated cognitive impairments, as accessed by the Passive avoidance and the Social Transfer of Food Preference (STFP) tests. Moreover, the numbers and densities of vascular and extracellular deposits containing amyloid beta peptide (Abeta) and carboxyl-terminal fragments of amyloid precursor protein (APP-CTFs), which are pathologic markers of AD, were significantly elevated in stressed animals, especially in the hippocampus. Moreover, stressed animals, also showed highly elevated levels of neurodegeneration and tau phosphorylation and increased intraneuronal Abeta and APP-CTFs immunoreactivities in the hippocampus and in the entorhinal and piriform cortex. This study provides the first evidence that chronic stress accelerates the onset and severity of cognitive deficits and that these are highly correlated with pathological changes, which thus indicates that chronic stress may be an important contributor to the onset and development of AD. PMID- 16467369 TI - Phospholipid transfer protein deficiency reduces sperm motility and impairs fertility of mouse males. AB - Vitamin E was discovered for its implication in reproductive biology, and its transport in mammalian plasma and brain was shown to be governed by plasma phospholipid transfer protein (PLTP). We show that PLTP deficiency is associated with hypofertility of mouse males but not mouse females, and it accounts for a significant decrease in total number of pups produced over a 2-month breeding period of PLTP knocked out mice (-32%, P<0.03). PLTP is highly expressed in epididymis of mouse males, and alpha-tocopherol, the main vitamin E isomer in vivo, was significantly less abundant in cauda and caput epididymis of PLTP deficient mice as compared with wild-type counterparts (caput: -26%, P<0.05; cauda: -21%, P<0.05). Mature spermatozoa from PLTP-deficient epididymis were shown to retain an abnormal alpha-tocopherol content. PLTP deficiency tended to reduce sperm motility as shown by a 24% reduction in spermatozoa with progressive motility (P<0.02), with no change in other sperm parameters as compared with wild type males. Finally, in vitro fertilization rates of wild-type oocytes with spermatozoa from PLTP-deficient males were markedly reduced as compared with those measured with spermatozoa from wild-type males (-60%, P<0.05). It is concluded that PLTP is a new, key factor that determines sperm motility and male fertility. PMID- 16467371 TI - Atorvastatin desensitizes beta-adrenergic signaling in cardiac myocytes via reduced isoprenylation of G-protein gamma-subunits. AB - Statins exert pleiotropic, cholesterol-independent effects by reducing isoprenylation of monomeric GTPases. Here we examined whether statins also reduce isoprenylation of gamma-subunits of heterotrimeric G-proteins and thereby affect beta-adrenergic signaling and regulation of force in cardiac myocytes. Neonatal rat cardiac myocytes (NRCM) were treated with atorvastatin (0.1-10 micromol/l; 12 48 h) and examined for adenylyl cyclase regulating G-protein alpha- (Galpha), beta- (Gbeta), and gamma- (Ggamma) subunits and cAMP accumulation. Engineered heart tissue (EHT) from NRCM was used to evaluate contractile consequences. In atorvastatin-treated NRCM, a second band of Ggamma3 with a lower apparent molecular weight appeared in cytosol and particulate fractions that was absent in vehicle-treated NRCM, but also seen after GGTI-298, a geranylgeranyl transferase inhibitor. In parallel, Gbeta accumulated in the cytosol and total cellular content of Galphas was reduced. In atorvastatin-treated NRCM, the cAMP-increasing effect of isoprenaline was reduced. Likewise, the positive inotropic effect of isoprenaline was desensitized and reduced after treatment with atorvastatin. The effects of atorvastatin were abolished by mevalonate and/or geranylgeranyl pyrophosphate, but not by farnesyl pyrophosphate or squalene. Taken together, the results of this study show that atorvastatin desensitizes NRCM to beta-adrenergic stimulation by a mechanism that involves reduced isoprenylation of Ggamma and subsequent reductions in the cellular content of Galphas. PMID- 16467372 TI - Real-time diagnostic imaging of tumors and metastases by use of a replication competent herpes vector to facilitate minimally invasive oncological surgery. AB - Current efforts on expanding minimally invasive techniques into the realm of oncological surgery are hindered by lack of accurate visualization of tumor margins and failure to detect micro metastases in real time. We used a systemic delivery of a herpes viral vector with cancer-selective infection and replication to precisely differentiate between normal and malignant tissue. NV1066 is a genetically modified, replication-competent herpes simplex virus carrying a transgene for enhanced green fluorescent protein (GFP). We tested the potential of NV1066 in delineating tumor tissue in vitro and in vivo in a wide range of cancers and whether NV1066-induced GFP expression can detect small foci of tumors and metastases in in vivo models using an operating endoscope with fluorescent filters. Our findings indicate that NV1066 can be used for real-time intraoperative imaging and enhanced detection of early cancers and metastases. We demonstrate that a single dose of NV1066, administered either locally (intratumoral or intracavitary) or systemically, will detect loco-regional and distant disease throughout the body. Such cancer selectivity is confirmed in 110 types of cancer cells from 16 different primary organs. Fluorescence-aided minimally invasive endoscopy revealed microscopic tumor deposits unrecognized by conventional laparoscopy/thoracoscopy. Furthermore, NV1066 ability to transit and infect tumor and metastases is proven in syngenic and transplanted tumors in different animal models, both immunocompetent and immunodeficient. Cancer selective GFP expression is confirmed by histology, immunohistochemistry, and qRT PCR. These studies form the basis for real-time, intraoperative diagnostic imaging of tumor and metastases by minimally invasive endoscopic technology. PMID- 16467373 TI - A PDZ-binding motif as a critical determinant of Rho guanine exchange factor function and cell phenotype. AB - We identified a Rho guanine exchange factor (GEF) expressed as two splice variants, which differ only in either having or lacking a Postsynaptic density 95, Disk large, Zona occludens-1 (PDZ) motif. The PDZ adaptor protein synectin bound the longer splice variant, Syx1, which was targeted to the plasma membrane in a synectin-dependent manner. The shorter variant, Syx2, was diffusely distributed in the cytoplasm. Fluorescence resonance energy transfer (FRET) imaging revealed similar differences between the spatial patterns of active RhoA in Syx1 versus Syx2-expressing cells. Expression of Syx1 augmented endothelial cell (EC) migration and tube formation, whereas Syx2 expression did not. It appears, therefore, that synectin-dependent targeting of Syx is critical to its contribution to these EC functions. Although agonist-stimulated global RhoA activity was similar in Syx1- and Syx2-expressing cells, basal RhoA activity was surprisingly higher in the latter. Out of 23 cell types, we found a significant level of endogenous Syx2 expression only in brain tumor cells, which also exhibited high basal RhoA activity. We found that the activity level of JNK, which mediates transcriptional regulation downstream of RhoA, is elevated in a Syx2-dependent manner in these cells, possibly contributing to their tumorigenicity. PMID- 16467374 TI - Role of LIM kinases in normal and psoriatic human epidermis. AB - We present evidence that LIM kinases can control cell adhesion and compaction in human epidermis. LIMK2 is expressed in the epidermal basal layer and signals downstream of the GTPase Rac1 to promote extracellular matrix adhesion and inhibit terminal differentiation. Conversely, LIMK1 is expressed in the upper granular layers and phosphorylates and inhibits cofilin. Expression of LIMK1 is lost in psoriatic lesions and other skin disorders characterized by lack of cell compaction in the differentiating cell layers. In psoriatic lesions down regulation of LIMK1 correlates with up-regulation of Myc. Expression of constitutively active cofilin or Myc in reconstituted human epidermis blocks cell compaction. Overexpression of LIMK1 leads to down-regulation of Myc, whereas inhibition of Rho kinase, an upstream activator of LIMK1, stimulates Myc expression. Inhibition of Myc by LIMK1 is via inhibition of Stat3 phosphorylation, because constitutively active cofilin or inhibition of Rho kinase results in Stat3 phosphorylation and increased Myc levels, whereas dominant negative Stat3 abolishes the effect. In conclusion, we have uncovered a novel antagonistic relationship between the LIMK1/phosphocofilin and Myc/Stat3 pathways in the differentiating layers of human epidermis and propose that down regulation of LIMK1 contributes to one of the pathological features of psoriatic epidermal lesions. PMID- 16467376 TI - Sp2 localizes to subnuclear foci associated with the nuclear matrix. AB - We have reported that extracts prepared from many human and mouse cell lines show little or no Sp2 DNA-binding activity and that Sp2 has little or no capacity to stimulate transcription of promoters that are activated by Sp1, Sp3, and Sp4. Using an array of chimeric Sp1/Sp2 proteins we showed further that Sp2 DNA binding activity and trans-activation are each negatively regulated in mammalian cells. As part of an ongoing effort to study Sp2 function and regulation we characterized its subcellular localization in comparison with other Sp-family members in fixed and live cells. We report that 1) Sp2 localizes largely within subnuclear foci associated with the nuclear matrix, and 2) these foci are distinct from promyelocytic oncogenic domains and appear to be stable during an 18-h time course of observation. Deletion analyses identified a 37 amino acid sequence spanning the first zinc-"finger" that is sufficient to direct nuclear matrix association, and this region also encodes a bipartite nuclear localization sequence. A second nuclear matrix targeting sequence is encoded within the Sp2 trans-activation domain. We conclude that Sp2 preferentially associates with the nuclear matrix and speculate that this subcellular localization plays an important role in the regulation of Sp2 function. PMID- 16467375 TI - CLIP-170 homologue and NUDE play overlapping roles in NUDF localization in Aspergillus nidulans. AB - Proteins in the cytoplasmic dynein pathway accumulate at the microtubule plus end, giving the appearance of comets when observed in live cells. The targeting mechanism for NUDF (LIS1/Pac1) of Aspergillus nidulans, a key component of the dynein pathway, has not been clear. Previous studies have demonstrated physical interactions of NUDF/LIS1/Pac1 with both NUDE/NUDEL/Ndl1 and CLIP-170/Bik1. Here, we have identified the A. nidulans CLIP-170 homologue, CLIPA. The clipA deletion did not cause an obvious nuclear distribution phenotype but affected cytoplasmic microtubules in an unexpected manner. Although more microtubules failed to undergo long-range growth toward the hyphal tip at 32 degrees C, those that reached the hyphal tip were less likely to undergo catastrophe. Thus, in addition to acting as a growth-promoting factor, CLIPA also promotes microtubule dynamics. In the absence of CLIPA, green fluorescent protein-labeled cytoplasmic dynein heavy chain, p150(Glued) dynactin, and NUDF were all seen as plus-end comets at 32 degrees C. However, under the same conditions, deletion of both clipA and nudE almost completely abolished NUDF comets, although nudE deletion itself did not cause a dramatic change in NUDF localization. Based on these results, we suggest that CLIPA and NUDE both recruit NUDF to the microtubule plus end. The plus-end localization of CLIPA itself seems to be regulated by different mechanisms under different physiological conditions. Although the KipA kinesin (Kip2/Tea2 homologue) did not affect plus-end localization of CLIPA at 32 degrees C, it was required for enhancing plus-end accumulation of CLIPA at an elevated temperature (42 degrees C). PMID- 16467378 TI - Interacting protein kinases involved in the regulation of flagellar length. AB - A striking difference of the life stages of the protozoan parasite Leishmania is a long flagellum in the insect stage promastigotes and a rudimentary organelle in the mammalian amastigotes. LmxMKK, a mitogen-activated protein (MAP) kinase kinase from Leishmania mexicana, is required for growth of a full-length flagellum. We identified LmxMPK3, a MAP kinase homologue, with a similar expression pattern as LmxMKK being not detectable in amastigotes, up-regulated during the differentiation to promastigotes, constantly expressed in promastigotes, and shut down during the differentiation to amastigotes. LmxMPK3 null mutants resemble the LmxMKK knockouts with flagella reduced to one-fifth of the wild-type length, stumpy cell bodies, and vesicles and membrane fragments in the flagellar pocket. A constitutively activated recombinant LmxMKK activates LmxMPK3 in vitro. Moreover, LmxMKK is likely to be directly involved in the phosphorylation of LmxMPK3 in vivo. Finally, LmxMPK3 is able to phosphorylate LmxMKK, indicating a possible feedback regulation. This is the first time that two interacting components of a signaling cascade have been described in the genus Leishmania. Moreover, we set the stage for the analysis of reversible phosphorylation in flagellar morphogenesis. PMID- 16467377 TI - Rad22Rad52-dependent repair of ribosomal DNA repeats cleaved by Slx1-Slx4 endonuclease. AB - Slx1 and Slx4 are subunits of a structure-specific DNA endonuclease that is found in Saccharomyces cerevisiae, Schizosaccharomyces pombe, and other eukaryotic species. It is thought to initiate recombination events or process recombination structures that occur during the replication of the tandem repeats of the ribosomal DNA (rDNA) locus. Here, we present evidence that fission yeast Slx1 Slx4 initiates homologous recombination events in the rDNA repeats that are processed by a mechanism that requires Rad22 (Rad52 homologue) but not Rhp51 (Rad51 homologue). Slx1 is required to generate approximately 50% of the spontaneous Rad22 DNA repair foci that occur in cycling cells. Most of these foci colocalize with the nucleolus, which contains the rDNA repeats. The increased fork pausing at the replication fork barriers in the rDNA repeats in a strain that lacks Rqh1 DNA helicase is further increased by expression of a dominant negative form of Slx1. These data suggest that Slx1-Slx4 cleaves paused replication forks in the rDNA, leading to Rad22-dependent homologous recombination that is used to maintain rDNA copy number. PMID- 16467380 TI - Regulated phosphorylation of budding yeast's essential myosin V heavy chain, Myo2p. AB - The tail of the yeast myosin V encoded by Myo2p is known to bind several receptors for cargo delivery along polarized actin cables. However, it is not known how Myo2p activity is regulated or how it selects between cargoes. Here we show that Myo2p is reversibly phosphorylated in vivo. A short peptide at the N terminal end of the cargo-binding domain contains three residues contributing to single or doubly phosphorylated species. We confirm that the tail consists of two proteolytically resistant subdomains and identify a functionally important region N-terminal to subdomain 1 that includes the phosphorylation sites. Mutagenesis of the phosphorylation sites to alanine abolished a mobility shift diagnostic of phosphorylation, whereas mutagenesis to glutamic acid produced the shift and the formation of an additional phosphorylated species. These substitutions did not affect overall cell growth. However, one of the sites is predicted to be a substrate of cAMP-dependent protein kinase (PKA), and yeast expressing Myo2p with alanine substitutions is resistant to otherwise lethal overexpression of PKA, whereas the glutamic acid mutant is supersensitive to overexpression of PKA. These results suggest that in yeast, Myo2p is subject to phosphoregulation involving a PKA-related signaling pathway. PMID- 16467379 TI - Actin-depolymerizing protein Adf1 is required for formation and maintenance of the contractile ring during cytokinesis in fission yeast. AB - The role of the actin-depolymerizing factor (ADF)/cofilin-family protein Adf1 in cytokinesis of fission yeast cells was studied. Adf1 was required for accumulation of actin at the division site by depolymerizing actin at the cell ends, assembly of the contractile ring through severing actin filaments, and maintenance of the contractile ring once formed. Genetic and cytological analyses suggested that it collaborates with profilin and capping protein in the mitotic reorganization of the actin cytoskeleton. Furthermore, it was unexpectedly found that Adf1 and myosin-II also collaborate in assembling the contractile ring. Tropomyosin was shown to antagonize the function of Adf1 in the contractile ring. We propose that formation and maintenance of the contractile ring are achieved by a balanced collaboration of these proteins. PMID- 16467381 TI - Protein kinase Czeta attenuates hypoxia-induced proliferation of fibroblasts by regulating MAP kinase phosphatase-1 expression. AB - We have previously found that hypoxia stimulates proliferation of vascular fibroblasts through Galphai-mediated activation of ERK1/2. Here, we demonstrate that hypoxia also activates the atypical protein kinase Czeta (PKCzeta) isozyme and stimulates the expression of ERK1/2-specific phosphatase, MAP kinase phosphatase-1 (MKP-1), which attenuates ERK1/2-mediated proliferative signals. Replication repressor activity is unique to PKCzeta because the blockade of classical and novel PKC isozymes does not affect fibroblast proliferation. PKCzeta is phosphorylated upon prolonged (24 h) exposure to hypoxia, whereas ERK1/2, the downstream kinases, are maximally activated in fibroblasts exposed to acute (10 min) hypoxia. However, PKCzeta blockade results in persistent ERK1/2 phosphorylation and marked increase in hypoxia-induced replication. Similarly prolonged ERK1/2 phosphorylation and increase in hypoxia-stimulated proliferation are also observed upon blockade of MKP-1 activation. Because of the parallel suppressive actions of PKCzeta and MKP-1 on ERK1/2 phosphorylation and proliferation, the role of PKCzeta in the regulation of MKP-1 expression was evaluated. PKCzeta attenuation reduces MKP-1 expression, whereas PKCzeta overexpression increases MKP-1 levels. In conclusion, our results indicate for the first time that hypoxia activates PKCzeta, which acts as a terminator of ERK1/2 activation through the regulation of downstream target, MKP-1 expression and thus serves to limit hypoxia-induced proliferation of fibroblasts. PMID- 16467382 TI - Yeast nuclear envelope subdomains with distinct abilities to resist membrane expansion. AB - Little is known about what dictates the round shape of the yeast Saccharomyces cerevisiae nucleus. In spo7Delta mutants, the nucleus is misshapen, exhibiting a single protrusion. The Spo7 protein is part of a phosphatase complex that represses phospholipid biosynthesis. Here, we report that the nuclear protrusion of spo7Delta mutants colocalizes with the nucleolus, whereas the nuclear compartment containing the bulk of the DNA is unaffected. Using strains in which the nucleolus is not intimately associated with the nuclear envelope, we show that the single nuclear protrusion of spo7Delta mutants is not a result of nucleolar expansion, but rather a property of the nuclear membrane. We found that in spo7Delta mutants the peripheral endoplasmic reticulum (ER) membrane was also expanded. Because the nuclear membrane and the ER are contiguous, this finding indicates that in spo7Delta mutants all ER membranes, with the exception of the membrane surrounding the bulk of the DNA, undergo expansion. Our results suggest that the nuclear envelope has distinct domains that differ in their ability to resist membrane expansion in response to increased phospholipid biosynthesis. We further propose that in budding yeast there is a mechanism, or structure, that restricts nuclear membrane expansion around the bulk of the DNA. PMID- 16467383 TI - H-Ras, R-Ras, and TC21 differentially regulate ureteric bud cell branching morphogenesis. AB - The collecting system of the kidney, derived from the ureteric bud (UB), undergoes repetitive bifid branching events during early development followed by a phase of tubular growth and elongation. Although members of the Ras GTPase family control cell growth, differentiation, proliferation, and migration, their role in development of the collecting system of the kidney is unexplored. In this study, we demonstrate that members of the R-Ras family of proteins, R-Ras and TC21, are expressed in the murine collecting system at E13.5, whereas H-Ras is only detected at day E17.5. Using murine UB cells expressing activated H-Ras, R Ras, and TC21, we demonstrate that R-Ras-expressing cells show increased branching morphogenesis and cell growth, TC21-expressing cells branch excessively but lose their ability to migrate, whereas H-Ras-expressing cells migrated the most and formed long unbranched tubules. These differences in branching morphogenesis are mediated by differential regulation/activation of the Rho family of GTPases and mitogen-activated protein kinases. Because most branching of the UB occurs early in development, it is conceivable that R-Ras and TC-21 play a role in facilitating branching and growth in early UB development, whereas H-Ras might favor cell migration and elongation of tubules, events that occur later in development. PMID- 16467384 TI - Cooperation between snail and LEF-1 transcription factors is essential for TGF beta1-induced epithelial-mesenchymal transition. AB - Transforming growth factor beta 1 (TGF-beta1) has been shown to induce epithelial mesenchymal transition (EMT) during various stages of embryogenesis and progressive disease. This alteration in cellular morphology is typically characterized by changes in cell polarity and loss of adhesion proteins such as E cadherin. Here we demonstrate that EMT is associated with loss of claudin-1, claudin-2, occludin, and E-cadherin expression within 72 h of exposure to TGF beta1 in MDCKII cells. It has been suggested that this expression loss occurs through TGF-beta1 in a Smad-independent mechanism, involving MEK and PI3K pathways, which have previously been shown to induce expression of the Snail (SNAI-1) gene. Here we show that these pathways are responsible for loss of tight junctions and a partial loss of E-cadherin. However, our results also demonstrate that a complete loss of E-cadherin and transformation to the mesenchymal phenotype are dependent on Smad signaling, which subsequently stimulates formation of beta-catenin/LEF-1 complexes that induce EMT. PMID- 16467385 TI - A role for PP1 in the Cdc2/Cyclin B-mediated positive feedback activation of Cdc25. AB - The Cdc25 phosphatase promotes entry into mitosis through the removal of inhibitory phosphorylations on the Cdc2 subunit of the Cdc2/CyclinB complex. During interphase, or after DNA damage, Cdc25 is suppressed by phosphorylation at Ser287 (Xenopus numbering; Ser216 of human Cdc25C) and subsequent binding of the small acidic protein, 14-3-3. As reported recently, at the time of mitotic entry, 14-3-3 protein is removed from Cdc25 and S287 is dephosphorylated by protein phosphatase 1 (PP1). After the initial activation of Cdc25 and consequent derepression of Cdc2/CyclinB, Cdc25 is further activated through a Cdc2-catalyzed positive feedback loop. Although the existence of such a loop has been appreciated for some time, the molecular mechanism for this activation has not been described. We report here that phosphorylation of S285 by Cdc2 greatly enhances recruitment of PP1 to Cdc25, thereby accelerating S287 dephosphorylation and mitotic entry. Moreover, we show that two other previously reported sites of Cdc2-catalyzed phosphorylation on Cdc25 are required for maximal biological activity of Cdc25, but they do not contribute to PP1 regulation and do not act solely through controlling S287 phosphorylation. Therefore, multiple mechanisms, including enhanced recruitment of PP1, are used to promote full activation of Cdc25 at the time of mitotic entry. PMID- 16467386 TI - Splicing speckles are not reservoirs of RNA polymerase II, but contain an inactive form, phosphorylated on serine2 residues of the C-terminal domain. AB - "Splicing speckles" are major nuclear domains rich in components of the splicing machinery and polyA(+) RNA. Although speckles contain little detectable transcriptional activity, they are found preferentially associated with specific mRNA-coding genes and gene-rich R bands, and they accumulate some unspliced pre mRNAs. RNA polymerase II transcribes mRNAs and is required for splicing, with some reports suggesting that the inactive complexes are stored in splicing speckles. Using ultrathin cryosections to improve optical resolution and preserve nuclear structure, we find that all forms of polymerase II are present, but not enriched, within speckles. Inhibition of polymerase activity shows that speckles do not act as major storage sites for inactive polymerase II complexes but that they contain a stable pool of polymerase II phosphorylated on serine(2) residues of the C-terminal domain, which is transcriptionally inactive and may have roles in spliceosome assembly or posttranscriptional splicing of pre-mRNAs. Paraspeckle domains lie adjacent to speckles, but little is known about their protein content or putative roles in the expression of the speckle-associated genes. We find that paraspeckles are transcriptionally inactive but contain polymerase II, which remains stably associated upon transcriptional inhibition, when paraspeckles reorganize around nucleoli in the form of caps. PMID- 16467387 TI - Kinesin-1 and Dynein are the primary motors for fast transport of mitochondria in Drosophila motor axons. AB - To address questions about mechanisms of filament-based organelle transport, a system was developed to image and track mitochondria in an intact Drosophila nervous system. Mutant analyses suggest that the primary motors for mitochondrial movement in larval motor axons are kinesin-1 (anterograde) and cytoplasmic dynein (retrograde), and interestingly that kinesin-1 is critical for retrograde transport by dynein. During transport, there was little evidence that force production by the two opposing motors was competitive, suggesting a mechanism for alternate coordination. Tests of the possible coordination factor P150(Glued) suggested that it indeed influenced both motors on axonal mitochondria, but there was no evidence that its function was critical for the motor coordination mechanism. Observation of organelle-filled axonal swellings ("organelle jams" or "clogs") caused by kinesin and dynein mutations showed that mitochondria could move vigorously within and pass through them, indicating that they were not the simple steric transport blockades suggested previously. We speculate that axonal swellings may instead reflect sites of autophagocytosis of senescent mitochondria that are stranded in axons by retrograde transport failure; a protective process aimed at suppressing cell death signals and neurodegeneration. PMID- 16467388 TI - Altered axonal excitability properties in amyotrophic lateral sclerosis: impaired potassium channel function related to disease stage. AB - Fasciculations are a characteristic feature of amyotrophic lateral sclerosis (ALS), and can arise proximally or distally in the motor neuron, indicating a widespread disturbance in membrane excitability. Previous studies of axonal excitability properties (i.e. threshold electrotonus, strength-duration time constant) have suggested respectively that change in potassium or sodium channels may be involved. To reinvestigate these changes and explore their correlation with disease stage, multiple axonal excitability properties (threshold electrotonus, strength-duration time constant, recovery cycle and current threshold relationship) were measured for the median nerve at the wrist in 58 ALS patients, and compared with 25 age-matched controls. In ALS, there were greater changes in depolarizing threshold electrotonus (i.e. less accommodation) (P < 0.001) and greater supernormality in the recovery cycles (P < 0.001). These abnormalities were more prominent in patients with moderately reduced CMAP (1-5 mV). Modelling the excitability changes in this group supported the hypothesis that axonal potassium conductances are reduced, resulting in increased supernormality despite membrane depolarization. The tendency for strength duration time constant to be prolonged in ALS was only significant for patients with normal CMAP amplitude (>5 mV). Patients with severely reduced CMAP (<1 mV) alone showed reduced threshold changes to hyperpolarizing current. These results suggest a changing pattern of abnormal membrane properties with disease progression. First, persistent Na+ conductance increases, possibly associated with collateral sprouting, and then K(+) conductances decline. Both changes cause axonal hyperexcitability, and may contribute to the generation of fasciculations. These serial changes in axonal properties could provide insights into the pathophysiology of ALS, and implications for future therapeutic options. PMID- 16467389 TI - Mecp2 deficiency is associated with learning and cognitive deficits and altered gene activity in the hippocampal region of mice. AB - Rett syndrome (RTT) is a debilitating neurological condition associated with mutations in the X-linked MECP2 gene, where apparently normal development is seen prior to the onset of cognitive and motor deterioration at 6-18 months of life. A targeted deletion of the methyl-CpG-binding domain (MBD) coding region and disruption of mRNA splicing was introduced in the mouse, resulting in a complete loss of Mecp2 transcripts and protein. Postnatal comparison of XO and XY mutant Mecp2 allele-containing null mice revealed similar effects on mouse growth and viability, suggesting that phenotypic manifestations are not modulated by the Y chromosome. Further assessment of Mecp2-null XY mice highlighted cerebellar and hippocampal/amygdala-based learning deficits in addition to reduced motor dexterity and decreased anxiety levels. Brain tissues containing the hippocampal formation of XY Mecp2-null mice also displayed significant changes in genetic activity, which are related to the severity of the mutant phenotype. PMID- 16467390 TI - Deep-breath frequency in bronchoconstricted monkeys (Macaca fascicularis). AB - Deep-breath frequency has been shown to increase in spontaneously obstructed asthmatic subjects. Furthermore, deep breaths are known to be regulated by lung rapidly adapting receptors, yet the mechanism by which these receptors are stimulated is unclear. This study tested the hypothesis that deep-breath frequency increases during experimentally induced bronchoconstriction, and the magnitude of the increased deep-breath frequency is dependent on the method by which bronchoconstriction is induced. Nine cynomolgus monkeys (Macaca fascicularis) were challenged with methacholine (MCh), Ascaris suum (AS), histamine, or an external mechanical resistance. Baseline (BL) and challenge deep breath frequency were calculated from the number of deep breaths per trial period. Airway resistance (Raw) and tissue compliance (Cti), as well as tidal volume, respiratory rate, and minute ventilation, were analyzed for BL and challenged conditions. Transfer impedance measurements were fit with the DuBois model to determine the respiratory parameters (Raw and Cti). The flow at the airway opening was measured and analyzed on a breath-by-breath basis to obtain the ventilatory parameters (tidal volume, respiratory rate, and minute ventilation). Deep-breath frequency resulting from AS and histamine challenges [0.370 (SD 0.186) and 0.467 breaths/min (SD 0.216), respectively] was significantly increased compared with BL, MCh, or external resistance challenges [0.61 (SD 0.046), 0.156 (SD 0.173), and 0.117 breaths/min (SD 0.082), respectively]. MCh and external resistance challenges resulted in insignificant changes in deep-breath frequency compared with BL. All four modalities produced similar levels of bronchoconstriction, as assessed through changes in Raw and Cti, and had similar effects on the ventilatory parameters except that non-deep breath tidal volume was decreased in AS and histamine. We propose that increased deep-breath frequency during AS and histamine challenge is the result of increased vascular permeability, which acts to increase rapidly adapting receptor activity. PMID- 16467392 TI - Perivascular nerves and the regulation of cerebrovascular tone. AB - Brain perfusion is tightly coupled to neuronal activity, is commonly used to monitor normal or pathological brain function, and is a direct reflection of the interactions that occur between neuronal signals and blood vessels. Cerebral blood vessels at the surface and within the brain are surrounded by nerve fibers that originate, respectively, from peripheral nerve ganglia and intrinsic brain neurons. Although of different origin and targeting distinct vascular beds, these "perivascular nerves" fulfill similar roles related to cerebrovascular functions, a major one being to regulate their tone and, therein, brain perfusion. This utmost function, which underlies the signals used in functional neuroimaging techniques and which can be jeopardized in pathologies such as Alzheimer's disease, stroke, and migraine headache, is thus regulated at several levels. Recently, new insights into our understanding of how neural input regulate cerebrovascular tone resulted in the rediscovery of the functional "neurovascular unit." These remarkable advances suggest that neuron-driven changes in vascular tone result from interactions that involve all components of the neurovascular unit, transducing neuronal signals into vasomotor responses not only through direct interaction between neurons and vessels but also indirectly via the perivascular astrocytes. Neurovascular coupling is thus determined by chemical signals released from activated perivascular nerves and astrocytes that alter vascular tone to locally adjust perfusion to the spatial and temporal changes in brain activity. PMID- 16467391 TI - No effect of menstrual cycle phase on glycerol or palmitate kinetics during 90 min of moderate exercise. AB - The systemic flux of glycerol and palmitate [a representative nonesterified free fatty acid (NEFA)] was assessed in three different phases of the menstrual cycle at rest and during moderate-intensity exercise. It was hypothesized that circulating glycerol and NEFA turnover would be greatest in the midfollicular (MF) phase of the menstrual cycle, when estrogen is elevated but progesterone low, followed by the midluteal phase (ML; high estrogen and progesterone), and lowest in the early follicular (EF) phase of the menstrual cycle (low estrogen and progesterone). Subjects included moderately active, eumenorrheic, healthy women. Testing occurred after 3 days of diet control and after an overnight fast (12-13 h). Resting and exercise (50% maximal oxygen uptake, 90 min) measurements of tracer-determined glycerol and palmitate kinetics were made. There was a significant increase in both glycerol and palmitate turnover from rest to exercise in all phases of the menstrual cycle (P<0.0001). No significant differences, however, were observed between cycle phases in the systemic flux of glycerol or palmitate, at rest or during exercise. Maximal peripheral lipolysis during exercise, as represented by glycerol rate of appearance at 90 min, equaled 8.45+/-0.96, 8.35+/-1.12, and 7.71+/-0.96 micromol.kg-1.min-1 in the EF, MF, and ML phases, respectively. Circulating free fatty acid utilization also peaked at 90 min of exercise, as indicated by the palmitate rate of disappearance (3.31+/ 0.35, 3.17+/-0.39, and 3.47+/-0.26 micromol.kg-1.min-1) in the EF, MF, and ML phases, respectively. In conclusion, systemic rates of glycerol and NEFA turnover (as represented by palmitate flux) were not significantly affected by the cyclic fluctuations in estrogen and progesterone that occur throughout the normal menstrual cycle, either at rest or during 90 min of moderate exercise. PMID- 16467394 TI - Point: supraspinal locomotor centers do contribute significantly to the hyperpnea of dynamic exercise. PMID- 16467396 TI - Comment on point: counterpoint "in health and in a normoxic environment, VO2 max is/is not limited primarily by cardiac output and locomotor muscle blood flow". Vol 100: 744-8, 2006. The major factor limiting VO2 max in health and normoxia is oxygen transport to active muscles. PMID- 16467397 TI - Lower body negative pressure vs. lower body positive pressure to prevent cardiac atrophy after bed rest and spaceflight. What caused the controversy? PMID- 16467393 TI - Carbon monoxide and hydrogen sulfide: gaseous messengers in cerebrovascular circulation. AB - This review focuses on two gaseous cellular messenger molecules, CO and H2S, that are involved in cerebrovascular flow regulation. CO is a dilatory mediator in active hyperemia, autoregulation, hypoxic dilation, and counteracting vasoconstriction. It is produced from heme by a constitutively expressed enzyme [heme oxygenase (HO)-2] expressed highly in the brain and by an inducible enzyme (HO-1). CO production is regulated by controlling substrate availability, HO-2 catalytic activity, and HO-1 expression. CO dilates arterioles by binding to heme that is bound to large-conductance Ca2+-activated K+ channels. This binding elevates channel Ca2+ sensitivity, that increases coupling of Ca2+ sparks to large-conductance Ca2+-activated K+ channel openings and, thereby, hyperpolarizes the vascular smooth muscle. In addition to dilating blood vessels, CO can either inhibit or accentuate vascular cell proliferation and apoptosis, depending on conditions. H2S may also function as a cerebrovascular dilator. It is produced in vascular smooth muscle cells by hydrolysis of l-cysteine catalyzed by cystathione gamma-lyase (CSE). H2S dilates arterioles at physiologically relevant concentrations via activation of ATP-sensitive K+ channels. In addition to dilating blood vessels, H2S promotes apoptosis of vascular smooth muscle cells and inhibits proliferation-associated vascular remodeling. Thus both CO and H2S modulate the function and the structure of circulatory system. Both the HO-CO and CSE-H2S systems have potential to interact with NO and prostanoids in the cerebral circulation. Much of the physiology and biochemistry of HO-CO and CSE H2S in the cerebral circulation remains open for exploration. PMID- 16467398 TI - Growth hormone responses to repeated bouts of aerobic exercise with different recovery intervals. PMID- 16467399 TI - IFNgamma pretreatment sensitizes human choriocarcinoma cells to etoposide-induced apoptosis. AB - Choriocarcinoma is a malignant trophoblast-derived tumour, which can arise in any type of gestation. Cell proliferation assays showed that interferon gamma (IFNgamma) alone significantly inhibited proliferation of choriocarcinoma JAR and JEG-3 cells. TdT (terminal deoxynucleotidyl transferase)-mediated dUDP nick-end labelling (TUNEL) assays and Hoechst staining indicated that IFNgamma alone could not induce apoptosis of JAR and JEG-3 cells, but IFNgamma could enhance the sensitivity of JAR cells to etoposide-induced apoptosis. RT-PCR and western blotting were performed to detect expression of apoptosis-related molecules IFNgammaR, interferon regulatory factor-1 (IRF-1), p53 and pro-caspase 3. In JAR cells, etoposide increased expression of the proteins including IFNgammaR, p53 and pro-caspase 3 as well as IRF-1 mRNA and IFNgamma-pretreatment apparently promoted up-regulation of these molecules expression. In addition, the responses of IRF-1, p53 and pro-caspase 3 expression to IFNgamma pretreatment were dose dependent. IRF-1 knock down assays demonstrated that IRF-1 directly mediated IFNgamma pretreatment enhanced sensitivity of JAR cells to etoposide-induced apoptosis and that pro-caspase 3 was one of the target genes of IRF-1. PMID- 16467400 TI - Stable gene silencing of synaptotagmin I in rat PC12 cells inhibits Ca2+-evoked release of catecholamine. AB - Synaptotagmin (syt) I is a Ca2+-binding protein that is well accepted as a major sensor for Ca2+-regulated release of transmitter. However, controversy remains as to whether syt I is the only protein that can function in this role and whether the remaining syt family members also function as Ca2+ sensors. In this study, we generated a PC12 cell line that continuously expresses a short hairpin RNA (shRNA) to silence expression of syt I by RNA interference. Immunoblot and immunocytochemistry experiments demonstrate that expression of syt I was specifically silenced in cells that stably integrate the shRNA-syt I compared with control cells stably transfected with the empty shRNA vector. The other predominantly expressed syt isoform, syt IX, was not affected, nor was the expression of the SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins when syt I levels were knocked down. Resting Ca2+ and stimulated Ca2+ influx imaged with fura-2 were not altered in syt I knockdown cells. However, evoked release of catecholamine detected by carbon fiber amperometry and HPLC was significantly reduced, although not abolished. Human syt I rescued the release events in the syt I knockdown cells. The reduction of stimulated catecholamine release in the syt I knockdown cells strongly suggests that although syt I is clearly involved in catecholamine release, it is not the only protein to regulate stimulated release in PC12 cells, and another protein likely has a role as a Ca2+ sensor for regulated release of transmitter. PMID- 16467401 TI - Serum deprivation results in redifferentiation of human umbilical vascular smooth muscle cells. AB - Phenotypic change of vascular smooth muscle cells (VSMCs) from a differentiated to a dedifferentiated state accompanies the early stage of atherosclerosis and restenosis. Although much progress has been made in determining the molecular mechanisms involved in VSMC dedifferentiation, research on VSMC redifferentiation is hindered by the lack of an appropriate complete redifferentiation model. We established an in vitro model of redifferentiation by using postconfluent VSMCs from human umbilical artery. We demonstrated that serum-deprived VSMCs are capable of complete redifferentiation. After serum deprivation, postconfluent cultured human umbilical VSMCs became elongated and spindle shaped, with elevation of myofilament density, and reacquired contraction. Expressions of VSMC specific contractile proteins, such as smooth muscle (SM) alpha-actin, SM-myosin heavy chain, calponin, and SM 22alpha, were increased and reached the levels in differentiated cells after serum deprivation. To determine the molecular mechanism of the phenotypic reversion, the levels of expression, phosphorylation, and binding activity of serum response factor (SRF), a key phenotypic modulator for VSMCs, were measured. The results showed that SRF binding activity with CArG motif was significantly increased after serum deprivation, whereas no changes were found in SRF expression and phosphorylation. The increased SRF binding activity was accompanied by an increase in expression of its coactivators such as myocardin. Furthermore, the phenotypic reversion was markedly inhibited by decoy double-strand oligodeoxynucleotides containing SM alpha-actin CArG motif, which was able to competitively bind to SRF. The results suggested that serum deprivation results in redifferentiation of human umbilical VSMCs. This novel model of VSMC phenotypic reversion should be valuable for research on vascular disease. PMID- 16467402 TI - The COX-2 pathway regulates growth of atrophied muscle via multiple mechanisms. AB - Loss of muscle mass occurs with disease, injury, aging, and inactivity. Restoration of normal muscle mass depends on myofiber growth, the regulation of which is incompletely understood. Cyclooxygenase (COX)-2 is one of two isoforms of COX that catalyzes the synthesis of prostaglandins, paracrine hormones that regulate diverse physiological and pathophysiological processes. Previously, we demonstrated that the COX-2 pathway regulates early stages of myofiber growth during muscle regeneration. However, whether the COX-2 pathway plays a common role in adult myofiber growth or functions specifically during muscle regeneration is unknown. Therefore, we examined the role of COX-2 during myofiber growth following atrophy in mice. Muscle atrophy was induced by hindlimb suspension (HS) for 2 wk, followed by a reloading period, during which mice were treated with either the COX-2-selective inhibitor SC-236 (6 mg x kg(-1) x day( 1)) or vehicle. COX-2 protein was expressed and SC-236 attenuated myofiber growth during reloading in both soleus and plantaris muscles. Attenuated myofiber growth in the soleus was associated with both decreased myonuclear addition and decreased inflammation, whereas neither of these processes mediated the effects of SC-236 on plantaris growth. In addition, COX-2(-/-) satellite cells exhibited impaired activation/proliferation in vitro, suggesting direct regulation of muscle cell activity by COX-2. Together, these data suggest that the COX-2 pathway plays a common regulatory role during various types of muscle growth via multiple mechanisms. PMID- 16467403 TI - Effect of gamma-secretase inhibitors on muscarinic receptor-mediated calcium signaling in human salivary epithelial cells. AB - Altered intracellular Ca(2+) signaling has been observed in cells derived from Alzheimer's disease patients, and a possible link between gamma-secretase activity and the content of intracellular Ca(2+) stores has been suggested. To test this hypothesis we studied the effects of several gamma-secretase inhibitors on muscarinic receptor-mediated intracellular calcium release in the human salivary gland cell line HSG. Although several inhibitors in the peptide aldehyde class partially blocked carbachol-induced Ca(2+) transients, these effects did not appear to be due to gamma-secretase inhibition, and overall we found no evidence that inhibition of gamma-secretase activity had any significant effect on agonist-induced intracellular calcium release in HSG cells. In complementary experiments with presenilin-null cells we found that the reconstitution of gamma secretase activity by transfection with wild-type presenilin 1 likewise had no significant effect on thapsigargin-induced Ca(2+) release. In a test of the specific hypothesis that the level of APP intracellular domain (AICD), the intracellular fragment of the beta-amyloid precursor protein (APP) resulting from gamma-secretase cleavage, can modulate the Ca(2+) content of the endoplasmic reticulum, we were unable to demonstrate any effect of APP small interfering RNA on the magnitude of carbachol-induced intracellular calcium release in HSG cells. Together our data cast considerable doubt on the hypothesis that there is a direct link between gamma-secretase activity and the content of intracellular Ca(2+) stores. PMID- 16467404 TI - CXCR3 chemokine receptor-induced chemotaxis in human airway epithelial cells: role of p38 MAPK and PI3K signaling pathways. AB - Human airway epithelial cells (HAEC) constitutively express the CXC chemokine receptor CXCR3, which regulates epithelial cell movement. In diseases such as chronic obstructive pulmonary disease and asthma, characterized by denudation of the epithelial lining, epithelial cell migration may contribute to airway repair and reconstitution. This study compared the potency and efficacy of three CXCR3 ligands, I-TAC/CXCL11, IP-10/CXCL10, and Mig/CXCL9, as inducers of chemotaxis in HAEC and examined the underlying signaling pathways involved. Studies were performed in cultured HAEC from normal subjects and the 16-HBE cell line. In normal HAEC, the efficacy of I-TAC-induced chemotaxis was 349 +/- 88% (mean +/- SE) of the medium control and approximately one-half the response to epidermal growth factor, a highly potent chemoattractant. In normal HAEC, Mig, IP-10, and I TAC induced chemotaxis with similar potency and a rank order of efficacy of I-TAC = IP-10 > Mig. Preincubation with pertussis toxin completely blocked CXCR3 induced migration. Of interest, intracellular [Ca(2+)] did not rise in response to I-TAC, IP-10, or Mig. I-TAC induced a rapid phosphorylation (5-10 min) of two of the three MAPKs, i.e., p38 and ERK1/2. Pretreatment of HAEC with the p38 inhibitor SB 20358 or the PI3K inhibitor wortmannin dose-dependently inhibited the chemotactic response to I-TAC. In contrast, the ERK1/2 inhibitor U0126 had no effect on chemotaxis. These data indicate that in HAEC, CXCR3-mediated chemotaxis involves a G protein, which activates both the p38 MAPK and PI3K pathways in a calcium-independent fashion. PMID- 16467405 TI - Matriptase activation and shedding with HAI-1 is induced by steroid sex hormones in human prostate cancer cells, but not in breast cancer cells. AB - Matriptase and its cognate inhibitor, hepatocyte growth factor activator inhibitor-1 (HAI-1), have been implicated in carcinoma onset and malignant progression. However, the pathological mechanisms of matriptase activation are not defined. Steroid sex hormones play crucial roles in prostate and breast cancer. Therefore, we investigated the questions of whether and how steroid sex hormones regulate matriptase activation in these cancer cells. Treatment of cells with 17beta-estradiol had no effect on activation of matriptase in hormone starved breast cancer cells, in part due to their high constitutive level of activated matriptase. In striking contrast, very low levels of activated matriptase were detected in hormone-starved lymph node prostatic adenocarcinoma (LNCaP) cells. Robust activation of matriptase was observed as early as 6 h after exposure of these cells to 5alpha-dihydrotestosterone (DHT). Activation of matriptase was closely followed by shedding of the activated matriptase with >90% of total activated matriptase present in the culture media 24 h after DHT treatment. Activated matriptase was shed in a complex with HAI-1 and may result from simultaneously proteolytic cleavages of both membrane-bound proteins. Latent matriptase and free HAI-1 were also shed into culture media. As a result of shedding, the cellular levels of matriptase and HAI-1 were significantly reduced 24 h after exposure to DHT. DHT-induced matriptase activation and shedding were significantly inhibited by the androgen antagonist bicalutamide, by the RNA transcription inhibitor actinomycin D, and by the protein synthesis inhibitor cycloheximide. These results suggest that in LNCaP cells, androgen induces matriptase activation via the androgen receptor, and requires transcription and protein synthesis. PMID- 16467406 TI - Combined acamprosate and naltrexone, with cognitive behavioural therapy is superior to either medication alone for alcohol abstinence: a single centres' experience with pharmacotherapy. AB - AIMS: To compare treatment outcomes amongst patients offered pharmacotherapy with either naltrexone or acamprosate used singly or in combination, in a 12-week outpatient cognitive behavioural therapy (CBT) programme for alcohol dependence. METHODS: We matched 236 patients across gender, age group, prior alcohol detoxification, and dependence severity and conducted a cohort comparison study of three medication groups (CBT + acamprosate, CBT + naltrexone, CBT + combined medication) which included 59 patients per group. Outcome measures included programme attendance, programme abstinence and for those who relapsed, cumulative abstinence duration (CAD) and days to first breach (DFB). Secondary analyses compared the remaining matched 59 subjects who declined medication with the pharmacotherapy groups. RESULTS: Across medication groups, CBT + combined medication produced the greatest improvement across all outcome measures. Although a trend favoured the CBT + combined group, differences did not reach statistical significance. Programme attendance: CBT + Acamprosate group (66.1%), CBT + Naltrexone group (79.7%), and in the CBT + Combined group (83.1%). Abstinence rates were 50.8, 66.1, and 67.8%, respectively. For those that did not complete the programme abstinent, the average number of days abstinent (CAD) were 45.07, 49.95, and 53.58 days, respectively. The average numbers of days to first breach (DFB) was 26.79, 26.7, and 37.32 days. When the focal group (CBT + combined) was compared with patients who declined medication (CBT-alone), significant differences were observed across all outcome indices. Withdrawal due to adverse medication effects was minimal. CONCLUSIONS: The addition of both medications (naltrexone and acamprosate) resulted in measurable benefit and was well tolerated. In this patient population naltrexone with CBT is as effective as combined medication with CBT, but the trend favours combination medication. PMID- 16467408 TI - alpha-Amylase and programmed cell death in aleurone of ripening wheat grains. AB - Late maturity alpha-amylase (LMA) in wheat is a genetic defect that may result in the accumulation of unacceptable levels of high pI alpha-amylase in grain in the absence of germination or weather damage. During germination, gibberellin produced in the embryo triggers expression of alpha-Amy genes, the synthesis of alpha-amylase and, subsequently, cell death in the aleurone. LMA also involves the aleurone and whilst LMA appears to be independent of the embryo there is nevertheless some evidence that gibberellin is involved. The aim of this investigation was to determine whether the increase in alpha-amylase activity in LMA-prone genotypes, like alpha-amylase synthesis by aleurone cells in germinating or GA-challenged grains, is followed by aleurone cell death. Programmed cell death was seen in aleurone layers from developing, ripe and germinated grains using confocal microscopy and fluorescent probes specific for dead or living cells. Small pockets of dying cells were observed distributed at random throughout the aleurone of ripening LMA-affected grains and by harvest ripeness these cells were clearly dead. The first appearance of dying cells, 35 d post-anthesis, coincided with the later part of the 'window of sensitivity' in grain development in LMA-prone wheat cultivars. No dead or dying cells were present in ripening or fully ripe grains of control cultivars. In germinating grains, dying cells were observed in the aleurone adjacent to the scutellum and, as germination progressed, the number of dead cells increased and the affected area extended further towards the distal end of the grain. Aside from the obvious differences in spatial distribution, dying cells in 20-24 h germinated grains were similar to dying cells in developing LMA-affected grains, consistent with previous measurements of alpha-amylase activity. The increase in high pI alpha amylase activity in developing grains of LMA-prone cultivars, like alpha-amylase synthesis in germinating grains, is associated with cell death, providing further evidence for the involvement of gibberellin in the LMA response. PMID- 16467407 TI - Physical training ameliorates chronic alcohol-induced hypertension and aortic reactivity in rats. AB - AIMS: The aim of this study was to investigate the interactive effect of physical training and chronic ethanol ingestion on changes in blood pressure (BP) and aortic reactivity response in rats. METHODS: Male Fisher rats were divided into four groups of seven animals each and treated as follows: (i) control (5% sucrose, orally) daily for 12 weeks; (ii) ethanol (4 g kg(-1), orally) daily for 12 weeks; (iii) exercise training on treadmill followed by sucrose daily for 12 weeks; (iv) exercise training on treadmill followed by ethanol daily for 12 weeks. The body weight and BP were recorded every week. The animals were anaesthetized with pentobarbital after 12 weeks; blood and thoracic aorta were isolated and analysed for ethanol and reactivity response using tissue bath technique, respectively. RESULTS: The data show that exercise training significantly lowered the weight gain 6-12 weeks in ethanol-treated rats compared to ethanol alone or control rats. The systolic and mean BP significantly elevated 6-12 weeks, whereas diastolic BP elevated 8-12 weeks after ethanol ingestion. Exercise training lowered the BP close to the normal control values in ethanol fed rats. Blood ethanol level significantly elevated in ethanol group but decreased in exercise plus alcohol group. Aortic contractile response to phenylephrine in ethanol or control groups was attenuated by training with or without intact endothelium. Ethanol significantly reduced the aortic relaxation response to acetylcholine whereas training enhanced the relaxation response with intact endothelium. The relaxation responses to adenosine and sodium nitroprusside in the aortic ring segments of rats with or without endothelium were decreased in ethanol group which were attenuated by exercise training. CONCLUSIONS: Physical training attenuates the chronic ethanol-induced hypertension via reduction of body weight, clearance of ethanol, and augmentation of the aortic endothelial relaxation response in rats. PMID- 16467409 TI - The effect of transient and continuous drought on yield, photosynthesis and carbon isotope discrimination in sugar beet (Beta vulgaris L.). AB - Stable carbon isotope discrimination (delta13C), photosynthetic performance (A), dry matter accumulation (DW), and sucrose yield (Y(s)) of sugar beet were evaluated in a glasshouse experiment under transient (TS) and permanent (PS) water stress. A was significantly reduced under drought, to an extent depending on stress duration. The reduced A was strictly associated with a low DW and Y(s), the later being 42% lower in PS than control plants (C). Restoring water steeply increased A and the associated leaf traits (RWC, leaf water potential etc.), but the increase of Y(s) was negligible. Therefore, the negative effects of severe water stress in the early growth period, though reversible on gas-exchange and most leaf traits, can drastically reduce Y(s) of sugar beet. Furthermore, A seems not to be effective in predicting sucrose accumulation, although it was very effective in detecting the occurrence of plant water stress. The A/C(i) model was used to assess the photosynthetic adjustments to continuous or transient drought by calculating the photosynthetic parameters Vcmax and Jmax and then compared with delta13C. Mesophyll conductance (g(m)) was estimated by comparing delta13C measured on soluble sugars and gas-exchange data. This approach confirmed the expectation that g(m) was limiting A and that there was a significant drop in [CO2] from the substomatal cavities and the chloroplast stroma both in favourable and drought conditions. Therefore, the carbon concentration at the carboxylation site was overestimated by 25-35% by conventional gas-exchange measurements, and Vcmax was consistently underestimated when g(m) was not taken into account, especially under severe drought. Root delta13C was found to be strictly related to sucrose content (brix%), Y(s) and root dry weight, and this was especially clear when delta13C was measured on bulk dry matter. By contrast, leaf delta13C measured in soluble sugars (delta(s)) and bulk dry matter (delta(dm)) were found to correlate weakly to brix% and yield, and this was not surprising as the integration time-scale of leaf delta(s) and delta(dm) were found to be shorter than that of root delta13C in bulk dry matter. The effect of water stress on diffusive and biochemical limitations with different integration times ranged from 1 d (leaf delta(s)) to more than 1 month (root delta(dm)). PMID- 16467410 TI - Salivary secretions by aphids interacting with proteins of phloem wound responses. AB - Successful phloem feeding requires overcoming a number of phloem-related plant properties and reactions. The most important hurdle is formed by the phloem wound responses, such as coagulating proteins in the phloem sieve elements of the plant and in the capillary food canal in the insect's mouth parts, i.e. the stylets. It seems that in order to prevent protein clogging inside a sieve element, ejection of watery saliva plays an important role. This ejection is detected in the electrical penetration graph (EPG) as E1 salivation and always precedes phloem sap ingestion. During this feeding from sieve elements, another regular and concurrent salivation also occurs, the watery E2 salivation. This E2 saliva is added to the ingested sap and, it probably prevents phloem proteins from clogging inside the capillary food canal. Whatever the biochemical mode of action of the inhibition of protein coagulation might be, in some plants aphids do not seem to be able to prevent clogging, which may explain the resistance to aphids in these plants. The relevance of this hypothesis is demonstrated by new experimental results and is related to new EPG results from plants with phloem-located resistance. PMID- 16467411 TI - Transfer of phloem-mobile substances from the host plants to the holoparasite Cuscuta sp. AB - During the development of the haustorium, searching hyphae of the parasite and the host parenchyma cells are connected by plasmodesmata. Using transgenic tobacco plants expressing a GFP-labelled movement protein of the tobacco mosaic virus, it was demonstrated that the interspecific plasmodesmata are open. The transfer of substances in the phloem from host to the parasite is not selective. After simultaneous application of (3)H-sucrose and (14)C-labelled phloem-mobile amino acids, phytohormones, and xenobiotica to the host, corresponding percentages of the translocated compounds are found in the parasite. An open continuity between the host phloem and the Cuscuta phloem via the haustorium was demonstrated in CLSM pictures after application of the phloem-mobile fluorescent probes, carboxyfluorescein (CF) and hydroxypyrene trisulphonic acid (HPTS), to the host. Using a Cuscuta bridge (14)C-sucrose and the virus PVY(N) were transferred from one host plant to the another. The results of translocation experiments with labelled compounds, phloem-mobile dyes and the virus should be considered as unequivocal evidence for a symplastic transfer of phloem solutes between Cuscuta species and their compatible hosts. PMID- 16467412 TI - Asymmetric distribution of auxin correlates with gravitropism and phototropism but not with autostraightening (autotropism) in pea epicotyls. AB - The relationships between the distribution of the native auxin indole-3-acetic acid (IAA) and tropisms in the epicotyl of red light-grown pea (Pisum sativum L.) seedlings have been investigated. The distribution measurement was made in a defined zone of the third internode, using (3)H-IAA applied from the plumule as a tracer. The tropisms investigated were gravitropism, pulse-induced phototropism, and time-dependent phototropism. The investigation was extended to the phase of autostraightening (autotropism) that followed gravitropic curvature. It was found that IAA is asymmetrically distributed between the two halves of the zone, with a greater IAA level occurring on the convex side, at early stages of gravitropic and phototropic curvatures. This asymmetry was found in epidermal peels and, except for one case (pulse-induced phototropism), no asymmetry was detected in whole tissues. It was concluded, in support of earlier results, that auxin asymmetry mediates gravitropism and phototropism and that the epidermis or peripheral cell layers play an important role in the establishment of auxin asymmetry in pea epicotyls. During autostraightening, which results from a reversal of growth asymmetry, the extent of IAA asymmetry was reduced, but its direction was not reversed. This result demonstrated that autostraightening is not regulated through auxin distribution. In this study, the growth on either side of the investigated zone was also measured. In some cases, the measured IAA distribution could not adequately explain the local growth rate, necessitating further detailed investigation. PMID- 16467413 TI - Common genetic variation in the prothrombin gene, hormone therapy, and incident nonfatal myocardial infarction in postmenopausal women. AB - Genetic variants in coagulation factors are associated with myocardial infarction and may modify the association between hormone therapy and cardiovascular disease risk. This study assessed whether common variation in the prothrombin gene was associated with incident nonfatal myocardial infarction in postmenopausal women and whether this association differed according to current estrogen use. Eight variants representing 98% of common prothrombin variants were selected using publicly available genomic variation data. These variants and the functional G20210A variant were genotyped and used to infer haplotypes in a population-based Washington State case-control study of postmenopausal Caucasian women (1995-1999; 273 cases and 788 controls). Women carrying a nonsynonymous polymorphism in exon 6 (C5467T) had an increased risk of myocardial infarction (for each additional copy, relative to women with one fewer copy, odds ratio = 1.4, 95% confidence interval: 1.0, 1.8). Prothrombin haplotypes were also associated with myocardial infarction (with minimal adjustment, global p = 0.056; with full adjustment, p = 0.034). Associations between haplotypes and myocardial infarction were similar among users of hormone therapy and nonusers (global p = 0.61), though statistical power was limited. These preliminary results suggest that common genetic variants in the prothrombin gene or other variants in linkage disequilibrium are associated with myocardial infarction in postmenopausal women. PMID- 16467414 TI - Occupational risk factors for esophageal and stomach cancers among female textile workers in Shanghai, China. AB - The authors evaluated associations between occupational exposures in the textile industry and the risks of esophageal cancer and stomach cancer. The authors conducted a case-cohort study nested in a cohort of female textile workers in Shanghai, China. One hundred and two workers with incident esophageal cancer and 646 workers with incident stomach cancer diagnosed between 1989 and 1998 were compared with an age-stratified reference subcohort (n = 3,188). Work histories were ascertained for all study subjects from factory personnel records or interviews. Exposures were reconstructed for chemicals and dusts by linking work history data with a job-exposure matrix developed for the Shanghai textile industry. Hazard ratios and 95 percent confidence intervals were calculated with Cox proportional hazards modeling adapted for the case-cohort design. Risk of esophageal cancer was associated with long-term (> or = 10 years) exposure to silica dust (hazard ratio = 15.8, 95% confidence interval: 3.5, 70.6) and metals (hazard ratio = 3.7, 95% confidence interval: 1.9, 7.1). Cumulative exposure to endotoxin, a contaminant of cotton dust, was inversely related to risks of both esophageal cancer (p-trend = 0.01) and stomach cancer (p-trend < 0.001) when exposures were lagged 20 years. Endotoxin has not been previously reported to be a protective factor for either stomach cancer or esophageal cancer and therefore warrants further study. PMID- 16467415 TI - Adult blood pressure and climate conditions in infancy: a test of the hypothesis that dehydration in infancy is associated with higher adult blood pressure. AB - The authors hypothesized that individuals born in the early 20th century who experienced the hottest and driest summers during infancy would be more likely to have suffered severe infant diarrhea and dehydration, and consequently have had higher blood pressure in adulthood, than those who experienced cooler and wetter summers. In this context, these climate data act as an instrumental variable for the association of early-life dehydration with later blood pressure. For 3,964 randomly selected British women born between 1919 and 1940 and whose blood pressure was measured at age 60-79 years, a one standard deviation (1.3 degrees C) higher mean summer temperature in the first year of life was associated with a 1.12-mmHg (95% confidence interval: 0.33, 1.91) higher adult systolic blood pressure, and a one standard deviation higher mean summer rainfall (33.9 mm) was associated with lower systolic blood pressure (-1.65 mmHg, 95% confidence interval: -2.44, -0.85). Equivalent results for diastolic blood pressure were 0.11 (95% confidence interval: -0.65, 0.86) and -0.32 (95% confidence interval: 0.71, 0.05). The climate variables were not associated with potential confounding factors such as socioeconomic position or lifestyle risk factors. These findings provide some evidence in favor of the hypothesis that dehydration in infancy is associated with higher adult blood pressure. PMID- 16467416 TI - Oral polio vaccine and intussusception: a data linkage study using records for vaccination and hospitalization. AB - The authors investigated the possibility of an association between oral polio vaccine (OPV) and intussusception by linking Scottish vaccination and hospitalization data sets and performing self-controlled case series analysis. The issue was important because rotavirus vaccine, another live oral virus vaccine, was withdrawn from the market in 1999 after studies showed a strong association with intussusception. OPV was recommended for all infants in the United Kingdom at ages 2, 3, and 4 months until 2004, when new combination vaccines containing inactivated poliovirus were introduced. Analysis was carried out for 466 intussusception cases occurring in 1987-1999 for which linked records on OPV vaccination were available. Six possible risk periods for intussusception, ranging from 3 days after vaccination to 41 days after vaccination, were examined, with separate analysis for each of the three OPV doses and also for data on all three doses combined. Of the 24 possible risk periods examined, the relative incidence of intussusception after vaccination was unchanged for 18, significantly decreased for five, and significantly increased for only one. The authors conclude that overall, there is no evidence for an association between OPV and intussusception, even when each dose is considered separately. PMID- 16467417 TI - Effects of sodium pump activity on spontaneous firing in neurons of the rat suprachiasmatic nucleus. AB - Cell-attached and whole cell recording techniques were used to study the effects of electrogenic sodium pump on the excitability of rat suprachiasmatic nucleus (SCN) neurons. Blocking the sodium pump with the cardiac steroid strophanthidin or zero K+ increased the spontaneous firing of SCN neurons to different degrees with different recording modes, whereas turning the sodium pump into a nonselective cation channel with the marine toxin palytoxin invariably increased the spontaneous firing to the point of total blockade. Current-clamp recordings indicated that strophanthidin increased the rate of membrane depolarization and reduced the peak afterhyperpolarization potential (AHP), whereas zero K+ also increased the rate of depolarization, but enhanced the peak AHP. The dual effect of zero K+ was reflected by the biphasic time course of voltage responses to zero K+: an inhibitory phase with enhanced peak AHP and slower firing, followed by a delayed excitatory phase with faster rate of membrane depolarization and faster firing. In the presence of strophanthidin to block the sodium pump, zero K+ consistently decreased firing by enhancing the peak AHP. Repetitive applications of K+ -free solution gradually turned the biphasic inhibitory-followed-by excitatory voltage response into a monophasic inhibitory response in cells recorded with the whole cell (but not the cell-attached) mode, suggesting rundown of sodium pump activity. Taken together, the results suggest that spontaneous firing of SCN neurons is regulated by sodium pump activity as well as the AHP, and that sodium pump activity is modulated by intracellular soluble substances subject to rundown under the whole cell conditions. PMID- 16467418 TI - Loss of TRPV1-expressing sensory neurons reduces spinal mu opioid receptors but paradoxically potentiates opioid analgesia. AB - Systemic administration of resiniferatoxin (RTX), an ultrapotent capsaicin analogue, removes transient receptor potential vanilloid type 1 (TRPV1) expressing afferent neurons and impairs thermal but not mechanical nociception in adult animals. In this study, we determined how loss of TRPV1-expressing sensory neurons alters the antinociceptive effect of mu opioids and mu opioid receptors in the spinal cord. The effect of morphine and (D-Ala2,N-Me-Phe4,Gly-ol5) enkephalin (DAMGO) was measured by testing the paw mechanical withdrawal threshold in rats treated with RTX or vehicle. RTX treatment deleted TRPV1 immunoreactive dorsal root ganglion neurons and nerve terminals in the spinal dorsal horn. Also the mu opioid receptor immunoreactivity was markedly reduced in the superficial dorsal horn of RTX-treated rats. However, RTX treatment did not affect the dorsal horn neurons labeled with both TRPV1- and mu opioid receptor immunoreactivity. Surprisingly, intrathecal morphine or DAMGO produced a greater increase in the withdrawal threshold in RTX- than in vehicle-treated rats. The duration of the effect of intrathecal morphine and DAMGO in RTX-treated rats was also profoundly increased. Furthermore, the antinociceptive effect of systemic morphine was significantly potentiated in RTX-treated rats. The B(MAX) (but not K(D)) of [3H]-DAMGO binding and DAMGO-stimulated [35S]GTPgammaS activity in the dorsal spinal cord were significantly reduced in the RTX group. This study provides novel information that loss of TRPV1 afferent neurons eliminates presynaptic mu opioid receptors present on TRPV1-expressing afferent neurons but paradoxically potentiates the analgesic effect of mu opioid agonists. Mechano nociception, transmitted through non-TRPV1 sensory neurons, is subject to potent modulation by mu opioid agonists. PMID- 16467419 TI - Nitric oxide transiently converts synaptic inhibition to excitation in retinal amacrine cells. AB - Nitric oxide (NO) is generated by multiple cell types in the vertebrate retina, including amacrine cells. We investigate the role of NO in the modulation of synaptic function using a culture system containing identified retinal amacrine cells. We find that moderate concentrations of NO alter GABA(A) receptor function to produce an enhancement of the GABA-gated current. Higher concentrations of NO also enhance GABA-gated currents, but this enhancement is primarily due to a substantial positive shift in the reversal potential of the current. Several pieces of evidence, including a similar effect on glycine-gated currents, indicate that the positive shift is due to an increase in cytosolic Cl-. This change in the chloride distribution is especially significant because it can invert the sign of GABA- and glycine-gated voltage responses. Furthermore, current- and voltage-clamp recordings from synaptic pairs of GABAergic amacrine cells demonstrate that NO transiently converts signaling at GABAergic synapses from inhibition to excitation. Persistence of the NO-induced shift in E(Cl-) in the absence of extracellular Cl- indicates that the increase in cytosolic Cl- is due to release of Cl- from an internal store. An NO-dependent release of Cl- from an internal store is also demonstrated for rat hippocampal neurons indicating that this mechanism is not restricted to the avian retina. Thus signaling in the CNS can be fundamentally altered by an NO-dependent mobilization of an internal Cl- store. PMID- 16467421 TI - Transient and steady-state dynamics of cortical adaptation. AB - Adaptation is a ubiquitous property of all sensory pathways of the brain and thus likely critical in the encoding of behaviorally relevant sensory information. Despite evidence identifying specific biophysical mechanisms contributing to sensory adaptation, its functional role in sensory encoding is not well understood, particularly in the natural environment where transient rather than steady-state activity could dominate the neuronal representation. Here, we show that the heterogeneous transient and steady-state adaptation dynamics of single cortical neurons in the rat vibrissa system were well characterized by an underlying state variable. The state was directly predictable from temporal response properties that capture the time course of postexcitatory suppression following an isolated vibrissa deflection. Altering the initial state, by preceding the periodic stimulus with an additional vibrissa deflection, strongly influenced single-cell transient cortical adaptation responses. Despite the different transient activity, neurons reached the same steady-state adapted response with a time to steady state that was independent of the initial state. However, the differences in transient activity observed on small time scales were not present when activity was integrated over the longer time scale of a stimulus cycle. Taken together, the results here demonstrate that although adaptation can have significant effects on transient neuronal activity and direction selectivity, a simple measure of the time course of suppression following an isolated stimulus predicted a large portion of the observed adaptation dynamics. PMID- 16467420 TI - Effect of pharmacological inactivation of nucleus reticularis tegmenti pontis on saccadic eye movements in the monkey. AB - The superior colliculus (SC) provides signals for the generation of saccades via a direct pathway to the brain stem burst generator (BG). In addition, it sends saccade-related activity to the BG indirectly through the cerebellum via a relay in the nucleus reticularis tegmenti pontis (NRTP). Lesions of the oculomotor vermis, lobules VIc and VII, and inactivation of the caudal fastigial nucleus, the cerebellar output nucleus to which it projects, produce saccade dysmetria but have little effect on saccade peak velocity and duration. We expected similar deficits from inactivation of the NRTP. Instead, injections as small as 80 nl into the NRTP first slowed ipsiversive saccades and then gradually reduced their amplitudes. Postinjection saccades had slower peak velocities and longer durations than preinjection saccades with similar amplitudes. Contraversive saccades retained their normal kinematics. When the gains of ipsiversive saccades to 10 degrees target steps had fallen to their lowest values (0.28 +/- 0.19; mean +/- SD; n = 10 experiments), the gains of contraversive saccades to 10 degrees target steps had decreased very little (0.82 +/- 0.11). Eventually, ipsiversive saccades did not exceed 5 degrees , even to 20 degrees target steps. Moreover, these small remaining saccades apparently were made with considerable difficulty because their latencies increased substantially. When ipsiversive saccade gain was at its lowest, the gain and kinematics of vertical saccades to 10 degrees target steps exhibited inconsistent changes. We argue that our injections did not compromise the direct SC pathway. Therefore these data suggest that the cerebellar saccade pathway does not simply modulate BG activity but is required for horizontal saccades to occur at all. PMID- 16467422 TI - Task switching as a two-stage decision process. AB - Saccades represent decisions, and the study of their latency has led to a neurally plausible model of the underlying mechanisms, LATER (Linear Approach to Threshold with Ergodic Rate), that can successfully predict reaction time behavior in simple decision tasks, with fixed instructions. However, if the instructions abruptly change, we have a more complex situation, known as task switching. Psychologists' explanations of the phenomena of task switching have so far tended to be qualitative rather than quantitative, and not intended to relate particularly clearly to existing models of decision making or to likely neural implementations. Here, we investigated task switching using a novel saccadic task: we presented the instructions by stimulus elements identical to those of the task itself, allowing us to compare decisions about instructions with decisions in the actual task. Our results support a relatively simple model consisting of two distinct LATER processes in series: the first detects the instruction, the second implements it. PMID- 16467423 TI - Selection and maintenance of saccade goals in the human frontal eye fields. AB - In a delayed-response task, response selection marks an important transition from sensory to motor processing. Using event-related functional magnetic resonance imaging, we imaged the human brain during performance of a novel delayed-saccade task that isolated response selection from visual encoding and motor execution. The frontal eye fields (FEFs) and intraparietal sulcus (IPS) both showed robust contra-lateralized activity time-locked to response selection. Moreover, response selection affected delay-period activity differently in these regions; it persisted throughout the memory delay period following response selection in the FEF but not IPS. Our results indicate that the FEF and IPS both make important but distinct contributions to spatial working memory. The mechanism that the FEF uses to support spatial working memory is tied to the selection and prospective coding of saccade goals, whereas the role of the IPS may be more tied to retrospective coding of sensory representations. PMID- 16467424 TI - Influence of spatial information on responses of tonically active neurons in the monkey striatum. AB - Previous studies have demonstrated that tonically active neurons (TANs) in the primate striatum play an important role in the detection of rewarding events. However, the influence of the spatial features of stimuli or actions required to obtain reward remains unclear. Here, we examined the activity of TANs in the striatum of monkeys trained to make spatially directed movements elicited by visual stimuli presented ipsilaterally or contralaterally to the moving arm. Among 181 neurons responding to the trigger stimulus, 127 (70%) were nonselective for stimulus location and 54 (30%) responded to only one location of the stimulus. Most of the selective responses (63%) occurred when the stimulus was presented contralaterally to the moving arm. To examine whether TAN responses are related to the location of the stimulus or to the direction of the movement, we tested a subset of the trigger-responsive neurons (n = 44) in a condition that elicited reaching toward or away from the stimulus. By comparing TAN activity between the two conditions, we found that half of the responses can be interpreted as being related to the location of the stimulus, one quarter to the direction of movement, and one quarter to the context in which stimulus-movement combination occurs. These results demonstrate that TANs are not limited to motivational processing, but may play a role in the processing of spatial attributes of stimulus and/or movement as well. These response properties suggest that TANs are involved in the flexible shifting of motor responses during spatially directed behavior. PMID- 16467425 TI - KCNQ/Kv7 channel regulation of hippocampal gamma-frequency firing in the absence of synaptic transmission. AB - Synchronous neuronal firing can be induced in hippocampal slices in the absence of synaptic transmission by lowering extracellular Ca2+ and raising extracellular K+. However, the ionic mechanisms underlying this nonsynaptic synchronous firing are not well understood. In this study we have investigated the role of KCNQ/Kv7 channels in regulating this form of nonsynaptic bursting activity. Incubation of rat hippocampal slices in reduced (<0.2 mM) [Ca2+]o and increased (6.3 mM) [K+]o, blocked synaptic transmission, increased neuronal firing, and led to the development of spontaneous periodic nonsynaptic epileptiform activity. This activity was recorded extracellularly as large (4.7 +/- 1.9 mV) depolarizing envelopes with superimposed high-frequency synchronous population spikes. These intraburst population spikes initially occurred at a high frequency (about 120 Hz), which decayed throughout the burst stabilizing in the gamma-frequency band (30-80 Hz). Further increasing [K+]o resulted in an increase in the interburst frequency without altering the intraburst population spike frequency. Application of retigabine (10 microM), a Kv7 channel modulator, completely abolished the bursts, in an XE-991-sensitive manner. Furthermore, application of the Kv7 channel blockers, linopirdine (10 microM) or XE-991 (10 microM) alone, abolished the gamma frequency, but not the higher-frequency population spike firing observed during low Ca2+/high K+ bursts. These data suggest that Kv7 channels are likely to play a role in the regulation of synchronous population firing activity. PMID- 16467426 TI - On the origin of the extracellular action potential waveform: A modeling study. AB - Although extracellular unit recording is typically used for the detection of spike occurrences, it also has the theoretical ability to report about what are typically considered intracellular features of the action potential. We address this theoretical ability by developing a model system that captures features of experimentally recorded simultaneous intracellular and extracellular recordings of CA1 pyramidal neurons. We use the line source approximation method of Holt and Koch to model the extracellular action potential (EAP) voltage resulting from the spiking activity of individual neurons. We compare the simultaneous intracellular and extracellular recordings of CA1 pyramidal neurons recorded in vivo with model predictions for the same cells reconstructed and simulated with compartmental models. The model accurately reproduces both the waveform and the amplitude of the EAPs, although it was difficult to achieve simultaneous good matches on both the intracellular and extracellular waveforms. This suggests that accounting for the EAP waveform provides a considerable constraint on the overall model. The developed model explains how and why the waveform varies with electrode position relative to the recorded cell. Interestingly, each cell's dendritic morphology had very little impact on the EAP waveform. The model also demonstrates that the varied composition of ionic currents in different cells is reflected in the features of the EAP. PMID- 16467427 TI - Ryanodine receptor regulates endogenous cannabinoid mobilization in the hippocampus. AB - Endogenous cannabinoids (eCBs) are produced and mobilized in a cytosolic calcium ([Ca2+]i)-dependent manner, and they regulate excitatory and inhibitory neurotransmitter release by acting as retrograde messengers. An indirect but real time bioassay for this process on GABAergic transmission is DSI (depolarization induced suppression of inhibition). The magnitude of DSI correlates linearly with depolarization-induced increase of [Ca2+]i that is thought to be initiated by Ca2+ influx through voltage-gated Ca2+ channels. However, the identity of Ca2+ sources involved in eCB mobilization in DSI remains undetermined. Here we show that, in CA1 pyramidal cells, DSI-inducing depolarizing voltage steps caused Ca2+ induced Ca2+ release (CICR) by activating the ryanodine receptor (RyR) Ca2+ release channel. CICR was reduced, and the remaining increase in [Ca2+]i was less effective in generating DSI, when the RyR antagonists, ryanodine or ruthenium red, were applied intracellularly, or the Ca2+ stores were depleted by the Ca2+ ATPase inhibitors, cyclopiazonic acid or thapsigargin. The CICR-dependent effects were most prominent in cultured or immature acute slices, but were also detectable in slices from adult tissue. Thus we suggest that voltage-gated Ca2+ entry raises local [Ca2+]i sufficiently to activate nearby RyRs and that the resulting CICR plays a critical role in initiating eCB mobilization. RyR may be a key molecule for the depolarization-induced production of eCBs that inhibit GABA release in the hippocampus. PMID- 16467428 TI - Guidance of eye movements during visual conjunction search: local and global contextual effects on target discriminability. AB - The composition of a visual scene influences the ability of humans to select specific details within that scene for discrimination or foveation with saccadic eye movements. With the goal of establishing an animal model to investigate the neural mechanisms underlying the deployment of visual attention and the guidance of saccades during visual search, we studied the visual behavior of three monkeys while they performed a conjunction (color + form) search task similar to those used in human studies. We found that search performance declined when distractors adjacent to the target shared its color, thereby revealing that color was more discriminable than form in these displays and suggesting that monkeys perceptually grouped stimuli by proximity and similarity. Search performance also varied with the overall composition of the display. Most importantly, saccades were biased toward distractors sharing the target color when there were few of them within the display and away from those distractors when they were numerous. Last, the monkeys initiated saccades with a fixed latency, suggesting that their responses to the display were automatic and that search strategies did not involve attentional resources beyond those recruited for regulating saccades. We conclude that monkeys adapt their visual strategies, largely via bottom-up processes, to both the local and the global context of the search. These findings suggest that the visual behavior of monkeys is guided by strategies similar to those observed in humans. PMID- 16467430 TI - Direct recording of theta oscillations in primate prefrontal and anterior cingulate cortices. AB - Recent evidence has suggested that theta-frequency (4-7 Hz) oscillations around the human anterior cingulate cortex (ACC) and frontal cortex--that is, frontal midline theta (Fm theta) oscillations--may be involved in attentional processes in the brain. However, little is known about the physiological basis of Fm theta oscillations because invasive study in the human is allowed in only limited cases. In the present study, we developed a monkey model for Fm theta oscillations and located the generators of theta waves using electrodes implanted in various cortical areas. Monkeys were engaged in a self-initiated hand-movement task with a waiting period. The theta power in area 9 (the medial prefrontal cortex) and area 32 (the rostral ACC) was gradually increased from a few seconds before the movement and reached a peak immediately after the movement. When the movement was rewarded, the theta power attained a second peak, whereas it swiftly decreased in the unrewarded trials. Theta oscillations in areas 9 and 32 were coherent and phase locked together. This theta activity may be associated with "executive attention" including self-control, internal timing, and assessment of reward. It is probably a homologue of human Fm theta oscillations, as judged from the similar localization, corresponding frequency, and dependency on attentional processes. The monkey model would be useful for studying executive functions in the frontal cortex. PMID- 16467429 TI - Stimulus-dependent changes in the vestibular contribution to human postural control. AB - Humans maintain stable stance in a wide variety of environments. This robust behavior is thought to involve sensory reweighting whereby the nervous system adjusts the relative contribution of sensory sources used to control stance depending on environmental conditions. Based on prior experimental and modeling results, we developed a specific quantitative representation of a sensory reweighting hypothesis that predicts that a given reduction in the contribution from one sensory system will be accompanied by a corresponding increase in the contribution from different sensory systems. The goal of this study was to test this sensory-reweighting hypothesis using measures that quantitatively assess the relative contributions of the proprioceptive and graviceptive (vestibular) systems to postural control during eyes-closed stance in different test conditions. Medial/lateral body sway was evoked by side-to-side rotation of the support surface (SS) while simultaneously delivering a pulsed galvanic vestibular stimulus (GVS) through electrodes behind the ears. A model-based interpretation of sway evoked by SS rotations provided estimates of the proprioceptive weighting factor, Wp, and showed that Wp declined with increasing SS amplitude. If the sensory-reweighting hypothesis is true, then the decline in Wp should be accompanied by a corresponding increase in Wp, the graviceptive weighting factor, and responses to the GVS should increase in proportion to the value of Wp derived from responses to SS rotations. Results were consistent with the predictions of the proposed sensory-reweighting hypothesis. GVS-evoked sway increased with increasing SS amplitude, and Wp measures derived from responses to GVS and from responses to SS rotations were highly correlated. PMID- 16467431 TI - Differentially variable component analysis: Identifying multiple evoked components using trial-to-trial variability. AB - Electric potentials and magnetic fields generated by ensembles of synchronously active neurons, either spontaneously or in response to external stimuli, provide information essential to understanding the processes underlying cognitive and sensorimotor activity. Interpreting recordings of these potentials and fields is difficult because detectors record signals simultaneously generated by various regions throughout the brain. We introduce a novel approach to this problem, the differentially variable component analysis (dVCA) algorithm, which relies on trial-to-trial variability in response amplitude and latency to identify multiple components. Using simulations we demonstrate the importance of response variability to component identification, the robustness of dVCA to noise, and its ability to characterize single-trial data. We then compare the source-separation capabilities of dVCA with those of principal component analysis and independent component analysis. Finally, we apply dVCA to neural ensemble activity recorded from an awake, behaving macaque-demonstrating that dVCA is an important tool for identifying and characterizing multiple components in the single trial. PMID- 16467432 TI - Layer-specific properties of the persistent sodium current in sensorimotor cortex. AB - We evaluated the characteristics of the persistent sodium current (I(NaP)) in pyramidal neurons of layers II/III and V in slices of rat sensorimotor cortex using whole cell patch-clamp recordings. In both layers, I(NaP) began activating around -60 mV and was half-activated at -43 mV. The I(NaP) peak amplitude and density were significantly higher in layer V. The voltage-dependent I(NaP) steady state inactivation occurred at potentials that were significantly more positive in layer V (V(1/2): -42.3 +/- 1.1 mV) than in layer II/III (V(1/2): -46.8 +/- 1.6 mV). In both layers, a current fraction corresponding to about 25% of the maximal peak amplitude did not inactivate. The time course of I(NaP) inactivation and recovery from inactivation could be fitted with a biexponential function. In layer V pyramidal neurons the faster time constant of development of inactivation had variable values, ranging from 158.0 to 1,133.8 ms, but it was on average significantly slower than that in layer II/III (425.9 +/- 80.5 vs. 145.8 +/- 18.2 ms). In both layers, I(NaP) did not completely inactivate even with very long conditioning depolarizations (40 s at -10 mV). Recovery from inactivation was similar in the two layers. Layer V intrinsically bursting and regular spiking nonadapting neurons showed particularly prolonged depolarized plateau potentials when Ca2+ and K+ currents were blocked and slower early phase of I(NaP) development of inactivation. The biexponential kinetics characterizing the time dependent inactivation of I(NaP) in layers II/III and V indicates a complex inactivating process that is incomplete, allowing a residual "persistent" current fraction that does not inactivate. Moreover, our data indicate that I(NaP) has uneven inactivation properties in pyramidal neurons of different layers of rat sensorimotor cortex. The higher current density, the rightward shifted voltage dependency of inactivation as well the slower kinetics of inactivation characterizing I(NaP) in layer V with respect to layer II/III pyramidal neurons may play a significant role in their ability to fire recurrent action potential bursts, as well in the high susceptibility to generate epileptic events. PMID- 16467433 TI - Olfactory nerve stimulation-evoked mGluR1 slow potentials, oscillations, and calcium signaling in mouse olfactory bulb mitral cells. AB - Fast synaptic transmission between olfactory receptor neurons and mitral cells (MCs) is mediated through AMPA and NMDA ionotropic glutamate receptors. MCs also express high levels of metabotropic glutamate receptor 1 (mGluR1) whose functional significance is less understood. Here we characterized a slow mGluR1 mediated potential that was evoked by high-frequency (100-Hz) olfactory nerve (ON) stimulation in the presence of NBQX and D-APV, blockers of ionotropic glutamate receptors, and that was associated with a local Ca2+ transient in the MC dendritic tuft. High-frequency ON stimulation in the presence of NBQX and D APV also evoked a slow, nearly 2-Hz oscillation of MC membrane potential that was abolished by the mGluR1 antagonist LY367385 (50 microM). Both mGluR slow potential and slow oscillation persisted in the presence of gabazine (10 microM), a GABA(A) receptor antagonist, and intracellular QX-314 (10 mM), a Na+ channel blocker. In contrast to a slow mGluR1 potential in cerebellar Purkinje neurons, the MC mGluR1 potential was not depressed by SKF96365 (< or =250 microM) and thus is likely not mediated by TRPC1 cation channels, nor was it potentiated by an elevation of intracellular Ca2+ level. Imaging with the Na+ indicator SBFI revealed a Na+ transient in the MC dendrite accompanying the mGluR1 slow potential. We conclude that the MC mGluR1 potential triggered by glutamate released from the ON supports oscillations and synchronizations of MCs associated within one glomerulus. PMID- 16467434 TI - Enzyme exposure in the British baking industry. AB - OBJECTIVES: Enzymes are commonly used in the baking industry, as they can improve dough quality and texture and lengthen the shelf life of the final product. There is little published information highlighting exposure to enzymes (other than fungal alpha-amylase) in the baking industry, therefore the purpose of this study was to identify antibodies and develop assays for the measurement of a variety of such enzymes in samples of airborne flour dust. METHODS: Polyclonal antibodies to bacterial amylase, glucose oxidase and amyloglucosidase were identified and developed into ELISA assays. The assays showed limited cross-reactivity with other enzymes commonly used in the baking industry. RESULTS: We measured levels of airborne enzymes in 195 personal air samples taken from a sample of 55 craft baking establishments. We were able to detect amyloglucosidase in 9% (16/184) of the samples, fungal alpha-amylase in 6% (11/171), bacterial alpha-amylase in 7% (13/195). However, we were unable to detect glucose oxidase in any of the samples. Measurements for protease enzymes were not carried out. Median levels in detectable samples of amyloglucosidase, fungal alpha-amylase and bacterial amylase were similar at 10.3, 5.3 and 5.9 ng/m(3), respectively. These figures represent the total enzyme protein (active and inactive) measured. CONCLUSIONS: There are few data in the literature regarding sensitization and exposure response relationships to these enzymes, and indeed there is often a lack of information within the industry as to the precise enzyme content of particular baking ingredients. As a precautionary measure, all enzymes are regarded as having the potential to cause respiratory sensitization. Consequently, exposures need to be controlled to as low a level as reasonably practicable, and future investigation may highlight the importance of measuring a variety of enzyme exposures and standardizing these methodologies to inform approaches to adequate control. PMID- 16467435 TI - Optimizing occupational exposure measurement strategies when estimating the log scale arithmetic mean value--an example from the reinforced plastics industry. AB - When assessing occupational exposures, repeated measurements are in most cases required. Repeated measurements are more resource intensive than a single measurement, so careful planning of the measurement strategy is necessary to assure that resources are spent wisely. The optimal strategy depends on the objectives of the measurements. Here, two different models of random effects analysis of variance (ANOVA) are proposed for the optimization of measurement strategies by the minimization of the variance of the estimated log-transformed arithmetic mean value of a worker group, i.e. the strategies are optimized for precise estimation of that value. The first model is a one-way random effects ANOVA model. For that model it is shown that the best precision in the estimated mean value is always obtained by including as many workers as possible in the sample while restricting the number of replicates to two or at most three regardless of the size of the variance components. The second model introduces the 'shared temporal variation' which accounts for those random temporal fluctuations of the exposure that the workers have in common. It is shown for that model that the optimal sample allocation depends on the relative sizes of the between-worker component and the shared temporal component, so that if the between-worker component is larger than the shared temporal component more workers should be included in the sample and vice versa. The results are illustrated graphically with an example from the reinforced plastics industry. If there exists a shared temporal variation at a workplace, that variability needs to be accounted for in the sampling design and the more complex model is recommended. PMID- 16467436 TI - The remarkable memory effects of propofol. PMID- 16467437 TI - Adjuncts to caudal blockade in children. PMID- 16467438 TI - Magnesium sulphate and ischaemic heart disease. PMID- 16467439 TI - Antiarrhythmic therapy and ECG. PMID- 16467440 TI - Novel use of an arterial cardiopulmonary bypass cannula as a tracheal tube. PMID- 16467441 TI - Laryngeal web as a result of Reinke's oedema: a cause of difficult endotracheal intubation. PMID- 16467442 TI - How much VEGF do you need? PMID- 16467443 TI - Foxp3-transduced polyclonal regulatory T cells protect against chronic renal injury from adriamycin. AB - Chronic proteinuric renal injury is a major cause of ESRD. Adriamycin nephropathy is a murine model of chronic proteinuric renal disease whereby chemical injury is followed by immune and structural changes that mimic human disease. Foxp3 is a gene that induces a regulatory T cell (Treg) phenotype. It was hypothesized that Foxp3-transduced Treg could protect against renal injury in Adriamycin nephropathy. CD4+ T cells were transduced with either a Foxp3-containing retrovirus or a control retrovirus. Foxp3-transduced T cells had a regulatory phenotype by functional and phenotypic assays. Adoptive transfer of Foxp3 transduced T cells protected against renal injury. Urinary protein excretion and serum creatinine were reduced (P<0.05), and there was significantly less glomerulosclerosis, tubular damage, and interstitial infiltrates (P<0.01). It is concluded that Foxp3-transduced Treg cells may have a therapeutic role in protecting against immune injury and disease progression in chronic proteinuric renal disease. PMID- 16467444 TI - Molecular comparison of calcineurin inhibitor-induced fibrogenic responses in protocol renal transplant biopsies. AB - The calcineurin inhibitor cyclosporine (CsA) induces a fibrogenic response that may lead to scarring of the renal allograft. This study investigated whether tacrolimus, a novel calcineurin inhibitor, exerts fibrogenic effects to a similar extent. Sixty patients were enrolled in a randomized study: 29 received CsA, and 31 received tacrolimus. Patients were subjected to tailored exposure-controlled calcineurin inhibitor regimens. Protocol biopsies were obtained at the time of transplantation and 6 and 12 mo after transplantation. Cortical TGF-beta and collagens alpha1(I) and alpha1(III) mRNA steady-state levels were determined with real-time PCR. The extent of protein deposition of TGF-beta, alpha-smooth muscle actin, and interstitial collagens in the renal cortex was quantified with computer-assisted image analysis. The extent of interstitial collagen deposition measured with Sirius red and the accumulation of alpha-smooth muscle actin and TGF-beta protein after 6 and 12 mo were similar for both immunosuppressive regimens. mRNA levels of TGF-beta and collagens alpha1(I) and alpha1(III) were not significantly different in the treatment groups either. It is concluded that the fibrogenic response in renal allografts is similar in patients who receive CsA-based regimens and patients who receive tacrolimus-based regimens. PMID- 16467445 TI - Bone marrow-derived cells do not contribute significantly to collagen I synthesis in a murine model of renal fibrosis. AB - Interstitial fibroblasts play a central role in kidney fibrosis. Their origin is debated, with recent data indicating a contribution of bone marrow (BM)-derived cells to the expanded population of interstitial cells after kidney damage in animals and humans. This study investigated whether these BM-derived cells would respond appropriately to a fibrotic drive by producing collagen. A transgenic mouse that expresses both luciferase and beta-galactosidase reporter molecules under the control of a 17-kb promoter and enhancer element of the gene encoding the alpha2 chain of the collagen I was used. Male transgenic BM was transplanted into female wild-type C57BL/6 mice (n=14), and unilateral ureteric obstruction was performed later to induce renal fibrosis. In the obstructed kidney of the BM chimeric female mice, a mean of 8.6% of smooth muscle actin-positive interstitial cells were Y chromosome positive. Increased collagen I mRNA in the obstructed kidney was detected by in situ hybridization. No luciferase activity was detected by enzyme assays in tissue homogenates of BM recipients, and very few luciferase mRNA transcripts were seen, mainly in tubular cells. beta-Galactosidase activity was not a useful reporter molecule because it could not be distinguished from enhanced endogenous beta-galactosidase activity in the obstructed kidney. These results indicate that BM-derived interstitial cells do not make a significant contribution to collagen I synthesis in the context of renal injury. PMID- 16467446 TI - Thin basement membrane nephropathy. PMID- 16467447 TI - Treatment with an inhibitory monoclonal antibody to mouse factor B protects mice from induction of apoptosis and renal ischemia/reperfusion injury. AB - Complement activation in the kidney after ischemia/reperfusion (I/R) seems to occur primarily via the alternative complement pathway. The ability of an inhibitory mAb to mouse factor B, a necessary component of the alternative pathway, to protect mice from ischemic acute renal failure was tested. Treatment with the mAb prevented the deposition of C3b on the tubular epithelium and the generation of systemic C3a after renal I/R. Treated mice had significantly lower increases in serum urea nitrogen and developed significantly less morphologic injury of the kidney after I/R. For gaining insight into potential mechanisms of protection, the activity of caspases within the kidney also was measured, and it was found that caspases-2, -3, and -9 increased in a complement-dependent manner after renal I/R. Apoptotic cells were detected by terminal deoxynucleotidyl transferase catalyzed labeling of DNA fragments, and mice in which the alternative pathway was inhibited demonstrated significantly less apoptosis than control mice. Thus, use of an inhibitory mAb to mouse factor B effectively prevented activation of complement in the kidney after I/R and protected the mice from necrotic and apoptotic injury of the tubules. PMID- 16467448 TI - Podocyte cell-specific expression of doxycycline inducible Cre recombinase in mice. AB - Conventional silencing of many podocyte-specific genes in mice is associated with embryonic or perinatal lethality. Therefore, it would be of great importance to generate mouse models that allow the modification of genes that are expressed in podocytes at later stages of age. Herein is described a transgenic mouse with doxycycline-inducible podocyte-specific expression of Cre recombinase. For the generation of this binary system, a single transgenic construct that contained two separate genes was used: One encoding the optimized M2 version of the doxycycline-dependent transcription transactivator reverse tetracycline controlled transcriptional activator (rtTA) under control of the human podocin (NPHS2) promoter and the other encoding the recombinase Cre under control of the rtTA/doxycycline-responsive minimal cytomegalovirus (CMV) Tet operator sequence 7 promotor. Microinjection of the JRC-CRE construct in fertilized oocytes from FVB/N mice resulted in 16 transgenic founders. Double-transgenic offspring from breeding of a selected founder with the Z/AP reporter mouse showed alkaline phosphatase staining only upon doxycycline administration and exclusively in podocytes. These data indicate that this new inducible Cre recombinase mouse line is an excellent tool in conditional, kidney glomerular podocyte-specific gene deletion in adult mice. PMID- 16467449 TI - Addition of the antioxidant probucol to angiotensin II type I receptor antagonist arrests progressive mesangioproliferative glomerulonephritis in the rat. AB - Angiotensin II (Ang II) and reactive oxidative species (ROS) that are produced by NADPH oxidase have been implicated in the progression of glomerulonephritis (GN). This study examined the effect of simultaneously interrupting Ang II and ROS with an Ang II receptor blocker (ARB), candesartan, and a free radical scavenger, probucol, in a model of progressive mesangioproliferative GN induced by the injection of anti-Thy-1 antibody into uninephrectomized rats. Nephritic rats were divided into four groups and given daily oral doses of the following: Vehicle, 1% probucol diet, 70 mg/L candesartan in drinking water, and probucol plus candesartan. These treatments lasted until day 56. Vehicle-treated nephritic rats developed progressively elevated proteinuria and glomerulosclerosis. Candesartan kept proteinuria significantly lower than vehicle or probucol. The addition of probucol to candesartan normalized urinary protein excretion. Increases in BP in nephritic rats were lowered by these treatments, except with probucol. It is interesting that both glomerular cell number and glomerulosclerosis were significantly decreased by candesartan and normalized by the addition of probucol. Immunohistochemical studies for TGF-beta1, collagen type I, and fibronectin revealed that the combined treatment abolished glomerular fibrotic findings compared with candesartan. In addition, glomerular expression of NADPH oxidase components and superoxide production suggested that the combined treatment completely eliminated NADPH oxidase-associated ROS production. In conclusion, our study provides the first evidence that the antioxidant probucol, when added to an Ang II receptor blockade, fully arrests proteinuria and disease progression in GN. Furthermore, the data suggest that NADPH oxidase-associated ROS production may play a pivotal role in the progression of GN. The combination of probucol and candesartan may represent a novel route of therapy for patients with progressive GN. PMID- 16467450 TI - A genome-wide DNA microsatellite association screen to identify chromosomal regions harboring candidate genes in diabetic nephropathy. AB - In an effort to accelerate the identification of susceptibility genes in diabetic nephropathy, the first genome-wide fluorescence-based DNA microsatellite (n=6000) association screen was performed, using pools of genomic DNA derived from Irish patients with (cases; n=200) and without (controls; n=200) type 1 diabetic nephropathy. Allele image profiles were generated for 5353 (89.2%) microsatellite markers for both case and control pools. Allele counts (estimated from allele image profiles) were compared in case versus control groups, and empirical P values were generated. Markers then were ranked on the basis of their empirical P values (lowest to highest). Repeat PCR amplification and electrophoresis of pooled samples were performed systematically on ranked markers until the 50 most associated markers with consistent results were identified. DNA samples that composed the pools then were genotyped individually for these markers. Two markers on chromosome 10, D10S558 (Pcorrected=0.005) and D10S1435 (Pcorrected=0.016), revealed statistically significant associations with diabetic nephropathy. An additional four markers (D6S281, D4S2937, D2S291, and D17S515) also are worthy of further investigation. Relevant functional candidate genes have been identified in the vicinity of these markers, demonstrating the feasibility of low-resolution genome-wide microsatellite association screening to identify possible candidate genes for diabetic nephropathy. PMID- 16467452 TI - Calcimimetic R-568 decreases extraosseous calcifications in uremic rats treated with calcitriol. AB - Calcimimetics decrease parathyroid hormone (PTH) levels in uremic patients with secondary hyperparathyroidism without increasing serum calcium (Ca). The aim of this study was to evaluate the effect of calcimimetic R-568 alone or in combination with calcitriol on vascular and other soft tissue calcifications in uremic rats with secondary hyperparathyroidism. Sham-operated and 5/6 nephrectomized Wistar rats were studied. 5/6 Nephrectomized rats were treated with vehicle, calcitriol (80 ng/kg every other day), R-568 (1.5 and 3 mg/kg per d), and both calcitriol and R-568 1.5 mg/kg, as above. Rats were killed after 14 or 56 d of treatment. Blood was drawn for biochemical measurements. Aortic, heart, kidney, lung, and stomach tissue samples were processed for histopathology and measurement of tissue Ca and phosphorus content. PTH concentrations were significantly reduced by all treatments. Treatment with calcitriol induced significant vascular calcification (aortic Ca increased to 4.2+/-1.2 mg/g at day 14 and to 11.4+/-0.7 mg/g at day 56; P<0.05 versus vehicle). Treatment with R-568 did not induce vascular calcification. Concurrent administration of R-568 with calcitriol reduced the aortic Ca (1.9+/-0.2 mg/g at day 14 and 7.5+/-1.4 mg/g at day 56) in relation to calcitriol alone. Soft tissue calcifications mirrored aortic mineralizations. Survival was significantly (P<0.001) reduced in calcitriol-treated rats, and mortality was attenuated (P=0.01) by concurrent treatment with R-568. In uremic rats, R-568 reduces elevated PTH levels without inducing vascular calcification, prevents calcitriol-induced vascular calcification, and decreases mortality. PMID- 16467451 TI - IL-6 haplotypes, inflammation, and risk for cardiovascular disease in a multiethnic dialysis cohort. AB - It is unknown whether IL-6, a central regulator of inflammation, is a cause of or just a marker of atherosclerosis. Studies of genetic susceptibility to inflammation, however, avoid the potential for reverse causality. Variation in IL6 gene was studied as a predictor of cardiovascular disease (CVD) risk in a cohort of 775 incident dialysis patients, in whom IL-6 levels are elevated. On the basis of published resequencing data on the IL6 gene, a phylogenetic tree with three main branches (clades 1 to 3) was constructed. Two "clade tag" polymorphisms, -174G/C and 1888G/T, and two missense variants, Pro32Ser and Asp162Val, were genotyped. Circulating IL-6 and albumin were measured a median of 5 mo after the start of dialysis. CVD events were ascertained from medical records. During a median follow-up of 2.5 yr, 294 CVD events occurred. The two coding variants, Pro32Ser (present only in black patients, 10% Ser allele) and Asp162Val (present only in white patients, 1% Val), were associated with lower levels of IL-6 and higher levels of albumin. The common variant in the promoter region, -174G/C, was strongly associated with higher CVD risk and weakly with IL 6 levels. Clade 3 (-174C carriers in the absence of 162 Val allele) was associated with higher IL-6 levels (P=0.03) and higher CVD risk (hazard ratio 1.44, P=0.006) after adjustment for covariates. The IL6 gene has functional variants that affect inflammation and risk for CVD among dialysis patients, supporting a causal role for IL6 in CVD. PMID- 16467453 TI - Attenuation of myocardial ischemia-reperfusion injury by trimetazidine derivatives functionalized with antioxidant properties. AB - Trimetazidine (TMZ), an anti-ischemic metabolic drug, is used to treat chest pain (angina pectoris). We hypothesized that derivatives of TMZ with antioxidant functions may improve the cardiac dysfunction caused by ischemia-reperfusion (I/R) above that observed with TMZ alone. Isolated rat hearts perfused with Krebs Henseleit buffer according to the Langendorff method were subjected to 30 min of global ischemia followed by 45 min of reperfusion. Trimetazidine, TMZ-NH (TMZ modified with a pyrroline moiety), or TMZ-PhiNH (TMZ-NH with a phenyl substitute) were infused (50 microM) for 1 min before the onset of ischemia. Untreated (control) hearts at the end of 45 min of reperfusion showed a significant decrease in the recovery of coronary flow (42%), left ventricular-developed pressure (22%), and rate-pressure product (25%) compared with preischemic baseline values. The I/R hearts also showed markedly increased lactate dehydrogenase and creatine kinase activities in the coronary effluent, significant myocardial infarction (46% of risk area), and activation of Akt, extracellular signal-regulated kinase, and p38 mitogen-activated protein kinase. Pretreatment of hearts with TMZ-NH or TMZ-PhiNH significantly enhanced the recovery of heart function and decreased infarct size. The I/R-induced activation of Akt was further enhanced by TMZ-PhiNH. The present study demonstrated that TMZ NH and TMZ-PhiNH significantly protected hearts against I/R-mediated cardiac dysfunction and injury. The protective effect of the TMZ derivatives could be due to the combined effects of antioxidant and anti-ischemic activities as well as enhanced pro-survival Akt activity. PMID- 16467454 TI - Expression profiles of high voltage-activated calcium channels in sympathetic and parasympathetic pelvic ganglion neurons innervating the urogenital system. AB - Among the autonomic ganglia, major pelvic ganglia (MPG) innervating the urogenital system are unique because both sympathetic and parasympathetic neurons are colocalized within one ganglion capsule. Sympathetic MPG neurons are discriminated from parasympathetic ones by expression of low voltage-activated Ca2+ channels that primarily arise from T-type alpha1H isoform and contribute to the generation of low-threshold spikes. Until now, however, expression profiles of high voltage-activated (HVA) Ca2+ channels in these two populations of MPG neurons remain unknown. Thus, in the present study, we dissected out HVA Ca2+ channels using pharmacological and molecular biological tools. Reverse transcription-polymerase chain reaction analysis showed that MPG neurons contained transcripts encoding all of the known HVA Ca2+ channel isoforms (alpha1B, alpha1C, alpha1D and alpha1E), with the exception of alpha1A. Western blot analysis and pharmacology with omega-agatoxin IVA (1 microM) confirmed that MPG neurons lack the alpha1A Ca2+ channels. Unexpectedly, the expression profile of HVA Ca2+ channel isoforms was identical in the sympathetic and parasympathetic neurons of the MPG. Of the total Ca2+ currents, omega-conotoxin GVIA-sensitive N type (alpha1B) currents constituted 57 +/- 5% (n = 9) and 60 +/- 3% (n = 6), respectively; nimodipine-sensitive L-type (alpha1C and alpha1D) currents made up 17 +/- 4% and 14 +/- 2%, respectively; and nimodipine-resistant and omega conotoxin GVIA-resistant R-type currents were 25 +/- 3% and 22 +/- 2%, respectively. The R-type Ca2+ currents were sensitive to NiCl2 (IC50 = 22 +/- 0.1 microM) but not to SNX-482, which was able to potently (IC50 = 76 +/- 0.4 nM) block the recombinant alpha1E/beta2a/alpha2delta Ca2+ currents expressed in human embryonic kidney 293 cells. Taken together, our data suggest that sympathetic and parasympathetic MPG neurons share a similar but unique profile of HVA Ca2+ channel isoforms. PMID- 16467455 TI - 3-[2-[4-(3-Chloro-2-methylphenylmethyl)-1-piperazinyl]ethyl]-5,6-dimethoxy-1-(4 imidazolylmethyl)-1H-indazole dihydro-chloride 3.5 hydrate (DY-9760e) is neuroprotective in rat microsphere embolism: role of the cross-talk between calpain and caspase-3 through calpastatin. AB - Microsphere embolism (ME)-induced cerebral ischemia can elicit various pathological events leading to neuronal death. Western blotting and immunohistochemical studies revealed that expression of calpastatin, an endogenous calpain inhibitor, decreased after ME induction. Calpain activation after ME was apparently due to, in part, a decrease in calpastatin in a late phase of neuronal injury. The time course of that decrease also paralleled caspase-3 activation. In vitro studies demonstrated that calpastatin was degraded by caspase-3 in a Ca(2+)/calmodulin (CaM)-dependent manner. Because CaM binds directly to calpastatin, we asked whether a novel CaM antagonist, 3-[2-[4-(3 chloro-2-methylphenylmethyl)-1-piperazinyl]ethyl]-5,6-dimethoxy-1-(4 imidazolylmethyl)-1H-indazole dihydro-chloride 3.5 hydrate (DY-9760e), inhibits caspase-3-induced calpastatin degradation during ME-induced neuronal damage. We also tested the effect of DY-9760e on degradation of fodrin, a calpain substrate. Consistent with our hypothesis, DY-9760e (25 or 50 mg/kg i.p.) treatment inhibited degradation of calpastatin and fodrin in a dose-dependent manner. Because DY-9760e showed powerful neuroprotective activity with concomitant inhibition of calpastatin degradation, cross-talk between calpain and caspase-3 through calpastatin possibly accounts for ME-induced neuronal injury. Taken together, both inhibition of caspase-3-induced calpastatin degradation and calpain-induced fodrin breakdown by DY-9760e in part mediate its neuroprotective action. PMID- 16467456 TI - Induction of hepatic transporters multidrug resistance-associated proteins (Mrp) 3 and 4 by clofibrate is regulated by peroxisome proliferator-activated receptor alpha. AB - Hepatic transporters play a vital role in the disposition of endogenous compounds and xenobiotics in the liver. The current study investigates the expression and regulation of hepatic efflux transporters in response to treatment with the peroxisome proliferator-activated receptor (PPAR)alpha agonist clofibrate (CFB). Changes in mRNA and protein levels for several hepatic transporters were assessed in male CD-1 mice after 10 days of CFB dosing (500 mg/kg i.p.). Administration of CFB up-regulated mRNA levels for breast cancer resistance protein (Bcrp) and multidrug resistance-associated proteins 3 and 4 (Mrp3 and Mrp4, respectively). Western blot analysis confirmed that CFB enhances protein expression of liver Bcrp, Mrp3, and Mrp4 in CD-1 mice. To further characterize the regulation of these hepatic transporters, CFB-mediated changes in transporter mRNA levels were assessed in wild-type (sv/129) and PPARalpha-null male mice. Wild-type mice treated with CFB showed similar changes in mRNA levels for all of these transporters, whereas the PPARalpha-null mice did not. Although protein expression of Mrp3 and Mrp4 in the wild-type mice correlated well with changes in mRNA levels, Bcrp protein was not up-regulated by CFB treatment. These results show that PPARalpha activation by CFB coordinately regulates the hepatic efflux transporters Mrp3 and Mrp4. Induction of Mrp3 and Mrp4 by CFB may alter the disposition of toxicants and xenobiotics that are substrates for these transporters. PMID- 16467457 TI - Can we trigger an acute coronary syndrome? PMID- 16467458 TI - Deprivation and trends in blood pressure, cholesterol, body mass index and smoking among participants of a UK primary care-based cardiovascular risk factor screening programme: both narrowing and widening in cardiovascular risk factor inequalities. AB - OBJECTIVES: To examine recent time trends in blood pressure (BP), total cholesterol, body mass index (BMI) and current smoking among people in the UK of different deprivation groups. DESIGN: Repeatable survey. SETTING: Primary care based UK cardiovascular risk factor screening programme (58 Stockport general practices). PARTICIPANTS: 37,161 women and 33,977 men aged 35-60 years responding to a screening invitation and with a first screening episode during 1989-99. RESULTS: There were significant decreasing trends in total cholesterol (-0.06 mmol/l/year, 95% confidence interval (CI) -0.07 to -0.06 for women, -0.07 mmol/l/year, 95% CI -0.07 to -0.06 for men), with a significantly faster drop in more deprived groups (-0.005 mmol/year/increasing deprivation group, 95% CI -0.01 to -0.001 for both sexes). There were decreasing trends in current smoking prevalence (odds ratio (OR) 0.97/year, 95% CI 0.96 to 0.97 for women, OR 0.96/year, 95% CI 0.95 to 0.96 for men) with a significantly slower drop in the more deprived groups (OR 1.01/year/increasing deprivation group, 95% CI 1.00 to 1.01 for both sexes). There were significant increasing trends in BMI (0.11 kg/m(2)/year in women, 95% CI 0.09 to 0.13, 0.10 kg/m(2)/year in men, 95% CI 0.08 to 0.11), with a significantly slower increase in the more deprived groups among men only (-0.02 kg/m(2)/year/increasing deprivation group, 95% CI -0.01 to 0.03). Inequality in BP narrowed among men but widened among women. CONCLUSION: Inequalities in risk factors between different deprivation groups may be both widening (smoking, BP in women) and narrowing (total cholesterol, BMI and BP in men). Given baseline inequalities in risk factors levels, these trends suggest that inequalities in cardiovascular disease are likely to persist in the future. PMID- 16467459 TI - Peak oxygen pulse during exercise as a predictor for coronary heart disease and all cause death. AB - OBJECTIVE: To investigate the prognostic value of peak oxygen pulse, which is the amount of oxygen consumed per heart beat during exercise, and to compare the prognostic value of peak oxygen pulse and maximum oxygen uptake (Vo(2max)) with respect to coronary heart disease (CHD) and overall death. DESIGN: Prospective population-based study based on 1596 men without CHD or the use of beta blockers at baseline. RESULTS: The risk of CHD was 2.45 (95% CI 1.10 to 5.45) times higher in men with low peak oxygen pulse (< 13.5 ml/beat) than in those with high peak oxygen pulse (> 17.8 ml/beat) after adjustment for age, alcohol consumption, smoking, body mass index, blood pressure, serum lipids, diabetes, family history of CHD and ischaemic ST changes during exercise. During an average follow up of 14 years, 267 men died, 67 of them due to CHD. The respective risk for overall death was 1.79 (95% CI 1.21 to 2.65). The continuous variable Vo(2max) was a stronger risk predictor than peak oxygen pulse for CHD and overall death. CONCLUSIONS: Assessment of oxygen pulse provides no complementary information to Vo(2max) about cardiorespiratory fitness and prognosis for CHD. The analysis of respiratory gas exchange including the assessment of oxygen pulse during exercise does, however, provide an additional means for defining prognosis for patients with CHD. PMID- 16467460 TI - Relationship between coronary flow velocity reserve and aortic stiffness. PMID- 16467461 TI - A novel mechanism for regulation of retinal blood flow by lactate: gap junctions, hypoxia, and pericytes. PMID- 16467462 TI - An emerging role for inflammatory cytokines in hypertension. PMID- 16467463 TI - The [PSI+] prion of Saccharomyces cerevisiae can be propagated by an Hsp104 orthologue from Candida albicans. AB - The molecular chaperone Hsp104 is not only a key component of the cellular machinery induced to disassemble aggregated proteins in stressed cells of Saccharomyces cerevisiae but also plays an essential role in the propagation of the [PSI+], [URE3], and [RNQ/PIN+] prions in this organism. Here we demonstrate that the fungal pathogen Candida albicans carries an 899-residue stress-inducible orthologue of Hsp104 (CaHsp104) that shows a high degree of amino acid identity to S. cerevisiae Hsp104 (ScHsp104). This identity is significantly lower in the N and C-terminal regions implicated in substrate recognition and cofactor binding, respectively. CaHsp104 is able to provide all known functions of ScHsp104 in an S. cerevisiae hsp104 null mutant, i.e., tolerance to high-temperature stress, reactivation of heat-denatured proteins, and propagation of the [PSI+] prion. As also observed for ScHsp104, overexpression of CaHsp104 leads to a loss of the [PSI+] prion. However, unlike that of ScHsp104, CaHsp104 function is resistant to guanidine hydrochloride (GdnHCl), an inhibitor of the ATPase activity of this chaperone. These findings have implications both in terms of the mechanism of inhibition of Hsp104 by GdnHCl and in the evolution of the ability of fungal species to propagate prions. PMID- 16467464 TI - Differential expression of Aspergillus nidulans ammonium permease genes is regulated by GATA transcription factor AreA. AB - The movement of ammonium across biological membranes is mediated in both prokaryotes and eukaryotes by ammonium transport proteins (AMT/MEP) that constitute a family of related sequences. We have previously identified two ammonium permeases in Aspergillus nidulans, encoded by the meaA and mepA genes. Here we show that meaA is expressed in the presence of ammonium, consistent with the function of MeaA as the main ammonium transporter required for optimal growth on ammonium as a nitrogen source. In contrast, mepA, which encodes a high affinity ammonium permease, is expressed only under nitrogen-limiting or starvation conditions. We have identified two additional AMT/MEP-like genes in A. nidulans, namely, mepB, which encodes a second high-affinity ammonium transporter expressed only in response to complete nitrogen starvation, and mepC, which is expressed at low levels under all nitrogen conditions. The MepC gene product is more divergent than the other A. nidulans AMT/MEP proteins and is not thought to significantly contribute to ammonium uptake under normal conditions. Remarkably, the expression of each AMT/MEP gene under all nitrogen conditions is regulated by the global nitrogen regulatory GATA factor AreA. Therefore, AreA is also active under nitrogen-sufficient conditions, along with its established role as a transcriptional activator in response to nitrogen limitation. PMID- 16467465 TI - MNN5 encodes an iron-regulated alpha-1,2-mannosyltransferase important for protein glycosylation, cell wall integrity, morphogenesis, and virulence in Candida albicans. AB - The cell walls of microbial pathogens mediate physical interactions with host cells and hence play a key role in infection. Mannosyltransferases have been shown to determine the cell wall properties and virulence of the pathogenic fungus Candida albicans. We previously identified a C. albicans alpha-1,2 mannosyltransferase, Mnn5, for its novel ability to enhance iron usage in Saccharomyces cerevisiae. Here we have studied the enzymatic properties of purified Mnn5 and characterized its function in its natural host. Mnn5 catalyzes the transfer of mannose to both alpha-1,2- and alpha-1,6-mannobiose, and this activity requires Mn2+ as a cofactor and is regulated by the Fe2+ concentration. An mnn5Delta mutant showed a lowered ability to extend O-linked, and possibly also N-linked, mannans, hypersensitivity to cell wall-damaging agents, and a reduction of cell wall mannosylphosphate content, phenotypes typical of many fungal mannosyltransferase mutants. The mnn5Delta mutant also exhibited some unique defects, such as impaired hyphal growth on solid media and attenuated virulence in mice. An unanticipated phenotype was the mnn5Delta mutant's resistance to killing by the iron-chelating protein lactoferrin, rendering it the first protein found that mediates lactoferrin killing of C. albicans. In summary, MNN5 deletion impairs a wide range of cellular events, most likely due to its broad substrate specificity. Of particular interest was the observed role of iron in regulating the enzymatic activity, suggesting an underlying relationship between Mnn5 activity and cellular iron homeostasis. PMID- 16467466 TI - Heterologous expression of membrane and soluble proteins derepresses GCN4 mRNA translation in the yeast Saccharomyces cerevisiae. AB - This paper describes the first physiological response at the translational level towards heterologous protein production in Saccharomyces cerevisiae. In yeast, the phosphorylation of eukaryotic initiation factor 2alpha (eIF-2alpha) by Gcn2p protein kinase mediates derepression of GCN4 mRNA translation. Gcn4p is a transcription factor initially found to be required for transcriptional induction of genes responsible for amino acid or purine biosynthesis. Using various GCN4 lacZ fusions, knockout yeast strains, and anti-eIF-2alpha-P/anti-eIF-2alpha antibodies, we observed that heterologous expression of the membrane-bound alpha1beta1 Na,K-ATPase from pig kidney, the rat pituitary adenylate cyclase seven-transmembrane-domain receptor, or a 401-residue soluble part of the Na,K ATPase alpha1 subunit derepressed GCN4 mRNA translation up to 70-fold. GCN4 translation was very sensitive to the presence of heterologous protein, as a density of 1 per thousand of heterologous membrane protein derepressed translation maximally. Translational derepression of GCN4 was not triggered by misfolding in the endoplasmic reticulum, as expression of the wild type or temperature-sensitive folding mutants of the Na,K-ATPase increased GCN4 translation to the same extent. In situ activity of the heterologously expressed protein was not required for derepression of GCN4 mRNA translation, as illustrated by the expression of an enzymatically inactive Na,K-ATPase. Two- to threefold overexpression of the highly abundant and plasma membrane-located endogenous H-ATPase also induced GCN4 translation. Derepression of GCN4 translation required phosphorylation of eIF-2alpha, the tRNA binding domain of Gcn2p, and the ribosome-associated proteins Gcn1p and Gcn20p. The increase in Gcn4p density in response to heterologous expression did not induce transcription from the HIS4 promoter, a traditional Gcn4p target. PMID- 16467467 TI - Role of N-terminal hydrophobic region in modulating the subcellular localization and enzyme activity of the bisphosphate nucleotidase from Debaryomyces hansenii. AB - 3', 5'-Bisphosphate nucleotidase is a ubiquitous enzyme that converts 3' phosphoadenosine-5'-phosphate to adenosine-5'-phosphate and inorganic phosphate. These enzymes are highly sensitive to sodium and lithium and, thus, perform a crucial rate-limiting metabolic step during salt stress in yeast. Recently, we have identified a bisphosphate nucleotidase gene (DHAL2) from the halotolerant yeast Debaryomyces hansenii. One of the unique features of Dhal2p is that it contains an N-terminal 54-amino-acid-residue hydrophobic extension. In this study, we have shown that Dhal2p exists as a cytosolic as well as a membrane bound form and that salt stress markedly influences the accumulation of the latter form in the cell. We have demonstrated that the N-terminal hydrophobic region was necessary for the synthesis of the membrane-bound isoform. It appeared that an alternative translation initiation was the major mechanism for the synthesis of these two forms. Moreover, the two forms exhibit significant differences in their substrate specificity. Unlike the cytosolic form, the membrane-bound form showed very high activity against inositol-1,4-bisphosphate. Thus, the present study for the first time reports the existence of multiple forms of a bisphosphate nucleotidase in any organism. PMID- 16467468 TI - Retention and loss of amino acid biosynthetic pathways based on analysis of whole genome sequences. AB - Plants and fungi can synthesize each of the 20 amino acids by using biosynthetic pathways inherited from their bacterial ancestors. However, the ability to synthesize nine amino acids (Phe, Trp, Ile, Leu, Val, Lys, His, Thr, and Met) was lost in a wide variety of eukaryotes that evolved the ability to feed on other organisms. Since the biosynthetic pathways and their respective enzymes are well characterized, orthologs can be recognized in whole genomes to understand when in evolution pathways were lost. The pattern of pathway loss and retention was analyzed in the complete genomes of three early-diverging protist parasites, the amoeba Dictyostelium, and six animals. The nine pathways were lost independently in animals, Dictyostelium, Leishmania, Plasmodium, and Cryptosporidium. Seven additional pathways appear to have been lost in one or another parasite, demonstrating that they are dispensable in a nutrition-rich environment. Our predictions of pathways retained and pathways lost based on computational analyses of whole genomes are validated by minimal-medium studies with mammals, fish, worms, and Dictyostelium. The apparent selective advantages of retaining biosynthetic capabilities for amino acids available in the diet are considered. PMID- 16467470 TI - A divergent transcription factor TFIIB in trypanosomes is required for RNA polymerase II-dependent spliced leader RNA transcription and cell viability. AB - Transcription by RNA polymerase II in trypanosomes deviates from the standard eukaryotic paradigm. Genes are transcribed polycistronically and subsequently cleaved into functional mRNAs, requiring trans splicing of a capped 39-nucleotide leader RNA derived from a short transcript, the spliced leader (SL) RNA. The only identified trypanosome RNA polymerase II promoter is that of the SL RNA gene. We have previously shown that transcription of SL RNA requires divergent trypanosome homologs of RNA polymerase II, TATA binding protein, and the small nuclear RNA (snRNA)-activating protein complex. In other eukaryotes, TFIIB is an additional key component of transcription for both mRNAs and polymerase II-dependent snRNAs. We have identified a divergent homolog of the usually highly conserved basal transcription factor, TFIIB, from the pathogenic parasite Trypanosoma brucei. T. brucei TFIIB (TbTFIIB) interacted directly with the trypanosome TATA binding protein and RNA polymerase II, confirming its identity. Functionally, in vitro transcription studies demonstrated that TbTFIIB is indispensable in SL RNA gene transcription. RNA interference (RNAi) studies corroborated the essential nature of TbTFIIB, as depletion of this protein led to growth arrest of parasites. Furthermore, nuclear extracts prepared from parasites depleted of TbTFIIB, after the induction of RNAi, required recombinant TbTFIIB to support spliced leader transcription. The information gleaned from TbTFIIB studies furthers our understanding of SL RNA gene transcription and the elusive overall transcriptional processes in trypanosomes. PMID- 16467469 TI - Copper induces cytoplasmic retention of fission yeast transcription factor cuf1. AB - Copper homeostasis within the cell is established and preserved by different mechanisms. Changes in gene expression constitute a way of maintaining this homeostasis. In Schizosaccharomyces pombe, the Cuf1 transcription factor is critical for the activation of copper transport gene expression under conditions of copper starvation. However, in the presence of elevated intracellular levels of copper, the mechanism of Cuf1 inactivation to turn off gene expression remains unclear. In this study, we provide evidence that inactivation of copper transport gene expression by Cuf1 is achieved through a copper-dependent, cytosolic retention of Cuf1. We identify a minimal nuclear localization sequence (NLS) between amino acids 11 to 53 within the Cuf1 N terminus. Deletion of this region and specific mutation of the Lys13, Arg16, Arg19, Lys24, Arg28, Lys45, Arg47, Arg50, and Arg53 residues to alanine within this putative NLS is sufficient to abrogate nuclear targeting of Cuf1. Under conditions of copper starvation, Cuf1 resides in the nucleus. However, in the presence of excess copper as well as silver ions, Cuf1 is sequestered in the cytoplasm, a process which requires the putative copper binding motif, 328Cys-X-Cys-X3-Cys-X-Cys-X2-Cys-X2-His342 (designated C-rich), within the C-terminal region of Cuf1. Deletion of this region and mutation of the Cys residues within the C-rich motif result in constitutive nuclear localization of Cuf1. By coexpressing the Cuf1 N terminus with its C terminus in trans and by using a two-hybrid assay, we show that these domains physically interact with each other in a copper-dependent manner. We propose a model wherein copper induces conformational changes in Cuf1 that promote a physical interaction between the Cuf1 N terminus and the C-rich motif in the C terminus that masks the NLS. Cuf1 is thereby sequestered in the cytosol under conditions of copper excess, thereby extinguishing copper transport gene expression. PMID- 16467472 TI - The function and properties of the Azf1 transcriptional regulator change with growth conditions in Saccharomyces cerevisiae. AB - Azf1 activates CLN3 transcription in Saccharomyces cerevisiae cells growing in glucose. Paradoxically, other studies have shown Azf1 to be almost undetectable in glucose-grown cells. Microarray experiments showed that Azf1 activates nonoverlapping gene sets in different carbon sources: in glucose, Azf1 activates carbon and energy metabolism genes, and in glycerol-lactate, Azf1 activates genes needed for cell wall maintenance. Consistent with the decreased expression of cell wall maintenance genes observed with azf1Delta mutants, we observed a marked growth defect in the azf1Delta cells at 37 degrees C in nonfermentable medium. Cell wall integrity assays, such as sensitivity to calcofluor white, sodium dodecyl sulfate, or caffeine, confirmed cell wall defects in azf1Delta mutants in nonfermentable medium. Gel shift experiments show that Azf1 binds to DNA elements with the sequence AAAAGAAA (A4GA3), a motif enriched in the promoters of Azf1 sensitive genes and predicted by whole-genome studies. This suggests that many of the Azf1-dependent transcripts may be regulated directly by Azf1 binding. We found that the levels of Azf1 protein in glucose-grown cells were comparable to Azf1 levels in cells grown in glycerol-lactate; however, this could only be demonstrated with a cell extraction procedure that minimizes proteolysis. Glucose produces conditions that destabilize the Azf1 protein, a finding that may reflect a glucose-induced change in Azf1 tertiary or quaternary structure. PMID- 16467471 TI - Ebs1p, a negative regulator of gene expression controlled by the Upf proteins in the yeast Saccharomyces cerevisiae. AB - Mutations in EBS1 were identified in Saccharomyces cerevisiae that cosuppress missense, frameshift, and nonsense mutations. Evidence from studies of loss of function and overexpression of EBS1 suggests that Ebs1p affects gene expression by inhibiting translation and that a loss of EBS1 function causes suppression by increasing the rate of translation. Changes in EBS1 expression levels alter the expression of wild-type genes, but, in general, no changes in mRNA abundance were associated with a loss of function or overexpression of EBS1. Translation of a lacZ reporter was increased in strains carrying an ebs1-Delta mutant gene, whereas translation was decreased when EBS1 was overexpressed. The cap binding protein eIF-4E copurifies with Ebs1p in the absence of RNA, suggesting that the two proteins interact in vivo. Although physical and genetic interactions were detected between Ebs1p and Dcp1p, copurification was RNase sensitive, and changes in the expression of Ebs1p had little to no effect on decapping of the MFA2 transcript. The combined results suggest that Ebs1p inhibits translation, most likely through effects on eIF-4E rather than on decapping. Finally, EBS1 transcript levels are under the control of nonsense-mediated mRNA decay (NMD), providing the first example of an NMD-sensitive transcript whose protein product influences a step in gene expression required for NMD. PMID- 16467473 TI - Rac1 and Cdc42 have different roles in Candida albicans development. AB - We investigated the role of the highly conserved G protein Rac1 in the opportunistic pathogen Candida albicans. We identified and disrupted RAC1 and show here that, in contrast to CDC42, it is not necessary for viability or serum induced hyphal growth but is essential for filamentous growth when cells are embedded in a matrix. Rac1 is localized to the plasma membrane, yet its distribution is more homogenous than that of Cdc42, with no enrichment at the tips of either buds or hyphae. In addition, fluorescence recovery after photobleaching results indicate that Rac1 and Cdc42 have different dynamics at the membrane. Furthermore, overexpression of Rac1 does not complement Cdc42 function, and conversely, overexpression of Cdc42 does not complement Rac1 function. Thus, Rac1 and Cdc42, although highly similar to one another, have different roles in C. albicans development. PMID- 16467475 TI - The Cek1 and Hog1 mitogen-activated protein kinases play complementary roles in cell wall biogenesis and chlamydospore formation in the fungal pathogen Candida albicans. AB - The Hog1 mitogen-activated protein (MAP) kinase mediates an adaptive response to both osmotic and oxidative stress in the fungal pathogen Candida albicans. This protein also participates in two distinct morphogenetic processes, namely the yeast-to-hypha transition (as a repressor) and chlamydospore formation (as an inducer). We show here that repression of filamentous growth occurs both under serum limitation and under other partially inducing conditions, such as low temperature, low pH, or nitrogen starvation. To understand the relationship of the HOG pathway to other MAP kinase cascades that also play a role in morphological transitions, we have constructed and characterized a set of double mutants in which we deleted both the HOG1 gene and other signaling elements (the CST20, CLA4, and HST7 kinases, the CPH1 and EFG1 transcription factors, and the CPP1 protein phosphatase). We also show that Hog1 prevents the yeast-to-hypha switch independent of all the elements analyzed and that the inability of the hog1 mutants to form chlamydospores is suppressed when additional elements of the CEK1 pathway (CST20 or HST7) are altered. Finally, we report that Hog1 represses the activation of the Cek1 MAP kinase under basal conditions and that Cek1 activation correlates with resistance to certain cell wall inhibitors (such as Congo red), demonstrating a role for this pathway in cell wall biogenesis. PMID- 16467474 TI - Genome-scale analysis reveals Sst2 as the principal regulator of mating pheromone signaling in the yeast Saccharomyces cerevisiae. AB - A common property of G protein-coupled receptors is that they become less responsive with prolonged stimulation. Regulators of G protein signaling (RGS proteins) are well known to accelerate G protein GTPase activity and do so by stabilizing the transition state conformation of the G protein alpha subunit. In the yeast Saccharomyces cerevisiae there are four RGS-homologous proteins (Sst2, Rgs2, Rax1, and Mdm1) and two Galpha proteins (Gpa1 and Gpa2). We show that Sst2 is the only RGS protein that binds selectively to the transition state conformation of Gpa1. The other RGS proteins also bind Gpa1 and modulate pheromone signaling, but to a lesser extent and in a manner clearly distinct from Sst2. To identify other candidate pathway regulators, we compared pheromone responses in 4,349 gene deletion mutants representing nearly all nonessential genes in yeast. A number of mutants produced an increase (sst2, bar1, asc1, and ygl024w) or decrease (cla4) in pheromone sensitivity or resulted in pheromone independent signaling (sst2, pbs2, gas1, and ygl024w). These findings suggest that Sst2 is the principal regulator of Gpa1-mediated signaling in vivo but that other proteins also contribute in distinct ways to pathway regulation. PMID- 16467477 TI - Physiological importance and identification of novel targets for the N-terminal acetyltransferase NatB. AB - The N-terminal acetyltransferase NatB in Saccharomyces cerevisiae consists of the catalytic subunit Nat3p and the associated subunit Mdm20p. We here extend our present knowledge about the physiological role of NatB by a combined proteomics and phenomics approach. We found that strains deleted for either NAT3 or MDM20 displayed different growth rates and morphologies in specific stress conditions, demonstrating that the two NatB subunits have partly individual functions. Earlier reported phenotypes of the nat3Delta strain have been associated with altered functionality of actin cables. However, we found that point mutants of tropomyosin that suppress the actin cable defect observed in nat3Delta only partially restores wild-type growth and morphology, indicating the existence of functionally important acetylations unrelated to actin cable function. Predicted NatB substrates were dramatically overrepresented in a distinct set of biological processes, mainly related to DNA processing and cell cycle progression. Three of these proteins, Cac2p, Pac10p, and Swc7p, were identified as true NatB substrates. To identify N-terminal acetylations potentially important for protein function, we performed a large-scale comparative phenotypic analysis including nat3Delta and strains deleted for the putative NatB substrates involved in cell cycle regulation and DNA processing. By this procedure we predicted functional importance of the N-terminal acetylation for 31 proteins. PMID- 16467476 TI - Candida albicans VAC8 is required for vacuolar inheritance and normal hyphal branching. AB - Hyphal growth is prevalent during most Candida albicans infections. Current cell division models, which are based on cytological analyses of C. albicans, predict that hyphal branching is intimately linked with vacuolar inheritance in this fungus. Here we report the molecular validation of this model, showing that a specific mutation that disrupts vacuolar inheritance also affects hyphal division. The armadillo repeat-containing protein Vac8p plays an important role in vacuolar inheritance in Saccharomyces cerevisiae. The VAC8 gene was identified in the C. albicans genome sequence and was resequenced. Homozygous C. albicans vac8Delta deletion mutants were generated, and their phenotypes were examined. Mutant vac8Delta cells contained fragmented vacuoles, and minimal vacuolar material was inherited by daughter cells in hyphal or budding forms. Normal rates of growth and hyphal extension were observed for the mutant hyphae on solid serum containing medium. However, branching frequencies were significantly increased in the mutant hyphae. These observations are consistent with a causal relationship between vacuolar inheritance and the cell division cycle in the subapical compartments of C. albicans hyphae. The data support the hypothesis that cytoplasmic volume, rather than cell size, is critical for progression through G1. PMID- 16467479 TI - Multifarious transcriptional regulation of adhesion protein gene ap65-1 by a novel Myb1 protein in the protozoan parasite Trichomonas vaginalis. AB - The transcription efficiency of an adhesion protein gene, ap65-1, in Trichomonas vaginalis varies with changes in the iron supply and with the growth stage. In the present study, two Myb recognition elements, MRE-1/MRE-2r and MRE-2f, were found to play antagonistic roles in regulating the iron-inducible activity of an ap65-1 reporter gene. Intriguingly, either of these elements was shown to be sufficient to repress basal activity, but together they were also shown to activate growth-related activity of the reporter gene in iron-depleted cells. A myb1 gene which encodes a 24-kDa protein containing a Myb-like R2R3 DNA binding domain was identified from Southwestern screening of MRE-2f-binding proteins. The Myb1 protein was detected as a major 35-kDa protein which exhibited variations in nuclear concentration with changes in the iron supply. A recombinant Myb1 protein was shown to differentially interact with MRE-1/MRE-2r and MRE-2f in vitro. Overexpression of hemagglutinin-tagged Myb1 in T. vaginalis resulted in repression or activation of ap65-1 transcription in iron-depleted cells at an early and a late stage of cell growth, respectively, while iron-inducible ap65-1 transcription was constitutively repressed. The hemagglutinin-tagged Myb1 protein was found to constantly occupy the chromosomal ap65-1 promoter at a proximal site, but it also selected two more distal sites only at the late growth stage. Together, these observations suggest that Myb1 critically regulates multifarious ap65-1 transcription, possibly via differential selection of multiple promoter sites upon environmental changes. PMID- 16467478 TI - The PbMDJ1 gene belongs to a conserved MDJ1/LON locus in thermodimorphic pathogenic fungi and encodes a heat shock protein that localizes to both the mitochondria and cell wall of Paracoccidioides brasiliensis. AB - J-domain (DnaJ) proteins, of the Hsp40 family, are essential cofactors of their cognate Hsp70 chaperones, besides acting as independent chaperones. In the present study, we have demonstrated the presence of Mdj1, a mitochondrial DnaJ member, not only in the mitochondria, where it is apparently sorted, but also in the cell wall of Paracoccidioides brasiliensis, a thermodimorphic pathogenic fungus. The molecule (PbMdj1) was localized to fungal yeast cells using both confocal and electron microscopy and also flow cytometry. The anti-recombinant PbMdj1 antibodies used in the reactions specifically recognized a single 55-kDa mitochondrial and cell wall (alkaline beta-mercaptoethanol extract) component, compatible with the predicted size of the protein devoid of its matrix peptide targeting signal. Labeling was abundant throughout the cell wall and especially in the budding regions; however, anti-PbMdj1 did not affect fungal growth in the concentrations tested in vitro, possibly due to the poor access of the antibodies to their target in growing cells. Labeled mitochondria stood preferentially close to the plasma membrane, and gold particles were detected in the thin space between them, toward the cell surface. We show that Mdj1 and the mitochondrial proteinase Lon homologues are heat shock proteins in P. brasiliensis and that their gene organizations are conserved among thermodimorphic fungi and Aspergillus, where the genes are adjacent and have a common 5' region. This is the first time a DnaJ member has been observed on the cell surface, where its function is speculative. PMID- 16467480 TI - The phosducin-like protein PhnA is required for Gbetagamma-mediated signaling for vegetative growth, developmental control, and toxin biosynthesis in Aspergillus nidulans. AB - Phosducin or phosducin-like protein (PhLP) is a positive regulator of Gbetagamma activity. The Gbeta (SfaD) and Ggamma (GpgA) subunits function in vegetative growth and developmental control in the model filamentous fungus Aspergillus nidulans. To better understand the nature of Gbetagamma-mediated signaling, phnA, encoding an A. nidulans PhLP, has been studied. Deletion of phnA resulted in phenotypes almost identical to those caused by deletion of sfaD, i.e., reduced biomass, asexual sporulation in liquid submerged culture, and defective fruiting body formation, suggesting that PhnA is necessary for Gbeta function. The requirement for the RGS protein FlbA in asexual sporulation could be bypassed by the DeltaphnA mutation, indicating that PhnA functions in FlbA-controlled vegetative growth signaling, primarily mediated by the heterotrimeric G protein composed of FadA (Galpha), SfaD, and GpgA. However, whereas deletion of fadA restored both asexual sporulation and the production of sterigmatocystin (ST), deletion of sfaD, gpgA, or phnA failed to restore ST production in the DeltaflbA mutant. Further studies revealed that SfaD, GpgA, and PhnA are necessary for the expression of aflR, encoding the transcriptional activator for the ST biosynthetic genes, and subsequent ST biosynthesis. Overexpression of aflR bypassed the need for SfaD in ST production, indicating that the results of SfaD mediated signaling may include transcriptional activation of aflR. Potential differential roles of FadA, Gbetagamma, and FlbA in controlling ST biosynthesis are further discussed. PMID- 16467481 TI - Vacuolar membrane dynamics in the filamentous fungus Aspergillus oryzae. AB - Vacuoles in filamentous fungi are highly pleomorphic and some of them, e.g., tubular vacuoles, are implicated in intra- and intercellular transport. In this report, we isolated Aovam3, the homologue of the Saccharomyces cerevisiae VAM3 gene that encodes the vacuolar syntaxin, from Aspergillus oryzae. In yeast complementation analyses, the expression of Aovam3 restored the phenotypes of both Deltavam3 and Deltapep12 mutants, suggesting that AoVam3p is likely the vacuolar and/or endosomal syntaxin in A. oryzae. FM4-64 [N-(3 triethylammoniumpropyl)-4-(p-diethylaminophenyl-hexatrienyl)pyridinium dibromide] and CMAC (7-amino-4-chloromethylcoumarin) staining confirmed that the fusion protein of enhanced green fluorescent protein (EGFP) with AoVam3p (EGFP-AoVam3p) localized on the membrane of the pleomorphic vacuolar networks, including large spherical vacuoles, tubular vacuoles, and putative late endosomes/prevacuolar compartments. EGFP-AoVam3p-expressing strains allowed us to observe the dynamics of vacuoles with high resolutions, and moreover, led to the discovery of several new aspects of fungal vacuoles, which have not been discovered so far with conventional staining methods, during different developmental stages. In old hyphae, EGFP fluorescence was present in the entire lumen of large vacuoles, which occupied most of the cell, indicating that degradation of cytosolic materials had occurred in such hyphae via an autophagic process. In hyphae that were not in contact with nutrients, such as aerial hyphae and hyphae that grew on a glass surface, vacuoles were composed of small punctate structures and tubular elements that often formed reticulum-like networks. These observations imply the presence of so-far-unrecognized roles of vacuoles in the development of filamentous fungi. PMID- 16467482 TI - Metallothionein gene from Tetrahymena thermophila with a copper-inducible repressible promoter. AB - We describe a novel metallothionein gene from Tetrahymena thermophila that has a strong copper-inducible promoter. This promoter can be turned on and off rapidly, making it a useful system for induction of ectopic gene expression in Tetrahymena and enhancing its applications in cell and molecular biology, as well as biotechnology. PMID- 16467483 TI - Variations on a theme. Regulation of flowering time in Arabidopsis. PMID- 16467484 TI - Ethics of medication-free research in schizophrenia. PMID- 16467485 TI - The origins, development, and passage of Medicare's revolutionary prospective payment system. AB - This article explains the origins, development, and passage of the single most influential postwar innovation in medical financing: Medicare's prospective payment system (PPS). Inexorably rising medical inflation and deep economic deterioration forced policymakers in the late 1970s to pursue radical reform of Medicare to keep the program from insolvency. Congress and the Reagan administration eventually turned to the one alternative reimbursement system that analysts and academics had studied more than any other and had even tested with apparent success in New Jersey: prospective payment with diagnosis-related groups (DRGs). Rather than simply reimbursing hospitals whatever costs they charged to treat Medicare patients, the new model paid hospitals a predetermined, set rate based on the patient's diagnosis. The most significant change in health policy since Medicare and Medicaid's passage in 1965 went virtually unnoticed by the general public. Nevertheless, the change was nothing short of revolutionary. For the first time, the federal government gained the upper hand in its financial relationship with the hospital industry. Medicare's new prospective payment system with DRGs triggered a shift in the balance of political and economic power between the providers of medical care (hospitals and physicians) and those who paid for it--power that providers had successfully accumulated for more than half a century. PMID- 16467486 TI - Two-week longitudinal survey of bone architecture alteration in the hindlimb unloaded rat model of bone loss: sex differences. AB - The goal of this study was to determine, through a longitudinal follow-up, whether sex influences bone adaptation during simulated weightlessness. Twelve week-old male and female Wistar rats were hindlimb unweighted for 2 wk, and the time course of bone alteration was monitored in vivo by means of densitometry and unbiased three-dimensional quantitative microcomputed tomography at 7 and 14 days. Compared with male rats, female rats had twice more cancellous bone volume at the proximal tibia at baseline, and this bone volume continued to increase, whereas in males it stabilized. Conversely, cortical area was greater in males than in females, and in both sexes cortical bone was still expanding. Hindlimb unloading resulted in larger reductions in males than in females in both cortical and cancellous compartments. In females, trabecular thickness and number decreased mildly, whereas in males trabecular number was dramatically reduced. In both sexes, the trabecular network became less connected and more rod-like shaped. Bone cellular activities evaluated by histomorphometry showed decreased bone formation rate in both sexes and increased resorption activity only in males. In conclusion, in female rats unloaded-related cancellous alterations reversed the growing process, whereas in males, which show lower growth process, it induced an accentuation of age-related cancellous bone changes for most of the parameters. PMID- 16467487 TI - Prolactin secretory rhythm of mated rats induced by a single injection of oxytocin. AB - Mating or vaginocervical stimulation [copulatory stimulus (CS)] induces two daily surges of the hormone prolactin (PRL) in rats. This unique secretory pattern of PRL surges is characteristic for the first half of pregnancy and is also present in ovariectomized (OVX) rats. Studies have shown that CS additionally provokes an acute release of the hormone oxytocin (OT). In this study, we tested whether a single injection of OT (iv) is sufficient to initiate the PRL secretion pattern of OVX/CS rats. Furthermore, we measured the 24-h profile of dopamine (DA) content in the anterior lobe of the pituitary gland, because DA is the major inhibitory factor of PRL secretion. The results indicated that a single injection of OT induces a PRL secretory rhythm and a DA release pattern similar to that initiated by CS. Immunocytochemical investigation showed that particular OTergic neurons in the hypothalamus express receptors for PRL, as well as for vasoactive intestinal polypeptide, which indicates an involvement in generating the PRL rhythm and entraining it to the ambient photoperiod. On the basis of this study, we suggest that the PRL-DA inhibitory feedback loop between lactotrophs and DAergic neurons plays a crucial role in generating the oscillatory PRL secretion pattern in CS rats. A timing signal, likely provided by the hypothalamic suprachiasmatic nucleus, entrains the autonomous PRL oscillation to a particular time of day. Mathematical modeling was used to illustrate the proposed network function. The experimental results further suggest an additional feedback mechanism in which certain hypothalamic OTergic neurons are influenced by PRL. PMID- 16467488 TI - A mathematical model for the mating-induced prolactin rhythm of female rats. AB - For the first 10 days of pregnancy and the first 12 days of pseudopregnancy, the secretion of prolactin (PRL) from pituitary lactotrophs is rhythmic, with two surges/day. This rhythm can also be triggered by bolus injection of oxytocin (OT). We describe a mathematical model for the initiation, maintenance, and termination of the OT-induced PRL rhythm. In our model, the mechanism for this circadian rhythm is mutual interaction between lactotrophs and neuroendocrine dopamine (DA) neurons. This rhythm is, under normal lighting conditions, entrained by the suprachiasmatic nucleus (SCN) but persists in the absence of input from the SCN. We postulate that OT injection triggers the rhythm by activating a population of bistable hypothalamic neurons that innervate and inhibit DA neurons. The bistable nature of these neurons allows them to act as a memory device, maintaining the rhythm long after OT has been cleared from the blood. The mechanism for this memory device and the arguments supporting it are detailed with computer simulations. Finally, we consider potential targets for a rhythm-terminating factor and make predictions that may be used to determine which mechanism is operational in terminating the OT- or mating-induced PRL rhythm. PMID- 16467489 TI - Proceedings of the SMBE Tri-National Young Investigators' Workshop 2005. Unusual evolution of interspersed repeat sequences in the Drosophila ananassae subgroup. AB - New repeat sequences were found in the Drosophila ananassae genome sequence. They accounted for approximately 1.2% of the D. ananassae genome and were estimated to be more abundant in genomes of its closely related species belonging to the Drosophila bipectinata complex, whereas it was entirely absent in the Drosophila melanogaster genome. They were interspersed throughout euchromatic regions of the genome, usually as short tandem arrays of unit sequences, which were mostly 175 200 bp long with two distinct peaks at 180 and 189 bp in the length distribution. The nucleotide differences among unit sequences within the same array (locus) were much smaller than those between separate loci, suggesting within-locus concerted evolution. The phylogenetic tree of the repeat sequences from different loci showed that divergences between sequences from different chromosome arms occurred only at earlier stages of evolution, while those within the same chromosome arm occurred thereafter, resulting in the increase in copy number. We found RNA polymerase III promoter sequences (A box and B box), which play a critical role in retroposition of short interspersed elements. We also found conserved stem-loop structures, which are possibly associated with certain DNA rearrangements responsible for the increase in copy number within a chromosome arm. Such an atypical combination of characteristics (i.e., wide dispersal and tandem repetition) may have been generated by these different transposition mechanisms during the course of evolution. PMID- 16467490 TI - Corticotropin-releasing hormone stimulates estrogen biosynthesis in cultured human placental trophoblasts. AB - Estrogens and corticotrophin-releasing hormone (CRH) produced by the placenta play pivotal roles in the control of parturition in human and other primates. There is a strong correlation between maternal CRH and estrogen concentrations throughout gestation. To investigate whether CRH produced locally in the placenta could modulate estrogen production, we obtained human placental trophoblasts from uncomplicated term pregnancies and cultured them for 72 h. Cells were then treated with CRH and with a CRH receptor antagonist, alpha-helical CRH9-41. The results showed that CRH stimulated, but alpha-helical CRH9-41 inhibited, the production of estradiol in a time- and dose-dependent manner. Consistent with this thesis, CRH increased whereas alpha-helical CRH decreased the mRNA levels of STS, CYP19A1, and HSD17B1, the key enzymes for estrogen synthesis. These results suggest that, in the placenta, endogenously produced CRH exhibits a tonic stimulatory effect on estrogen production. PMID- 16467491 TI - Identification of rat cysteine-rich secretory protein 4 (Crisp4) as the ortholog to human CRISP1 and mouse Crisp4. AB - Cysteine-rich secretory proteins (CRISPs) are present in a diverse population of organisms and are defined by 16 conserved cysteine residues spanning a plant pathogenesis related-1 and a C-terminal cysteine-rich domain. To date, the diversification of mammalian CRISPs is evidenced by the existence of two, three, and four paralogous genes in the rat, human, and mouse, respectively. The current study identifies a third rat Crisp paralog we term Crisp4. The gene for Crisp4 is on rat chromosome 9 within 1 Mb of both the Crisp1 and Crisp2 genes. The full length transcript for this gene was cloned from rat epididymal RNA and encodes a protein that shares 69% and 91% similarity with human CRISP1 and mouse CRISP4, respectively. Expression of rat Crisp4 is most abundant in the epididymis, with the highest levels of transcription observed in the caput and corpus epididymis. In contrast, rat CRISP4 protein is most abundant in the corpus and cauda regions of the epididymis. Rat CRISP4 protein is also present in caudal sperm extracts, appearing as a detergent-soluble form at the predicted MWR (26 kDa). Our data identify rat Crisp4 as the true ortholog to human CRISP1 and mouse Crisp4, and demonstrate its interaction with spermatozoa in the epididymis. PMID- 16467492 TI - Expression of the insulin-like peptide 3 (INSL3) hormone-receptor (LGR8) system in the testis. AB - The new peptide hormone insulin-like peptide 3 (INSL3) is a member of the insulin relaxin family, yet, unlike insulin, it signals through a new G-protein coupled receptor, LGR8, distantly related to the receptors for LH and FSH. INSL3 is produced in large amounts by the Leydig cells of the testis in both fetal and adult mammals. Using a combination of mRNA analysis by RT-PCR, immunohistochemistry, ligand-binding, and/or bioactivity assays, the distribution of LGR8 expression was assessed in testicular tissues and cells and in the epididymis. There was consistent agreement that LGR8 was expressed in meiotic and particularly postmeiotic germ cells and in Leydig cells, though not in Sertoli or peritubular cells. Leydig cells appear to express only a low level of the LGR8 gene product; other transcripts may be present, representing nonfunctional products. Messenger RNA analysis suggested that LGR8 transcripts in germ cells represented mostly full-length forms. LGR8 mRNA was also expressed in the epididymis, though no function can yet be ascribed to this expression. Therefore, the INSL3/LGR8 system represents a further paracrine hormone-receptor system in the testis, which conveys information about Leydig cell status to germ cells, and possibly as part of an autocrine feedback loop. PMID- 16467493 TI - Leisure activities and the risk of amnestic mild cognitive impairment in the elderly. AB - OBJECTIVE: To study the influence of leisure activity participation on risk of development of amnestic mild cognitive impairment (aMCI). METHODS: The authors examined the relationship between baseline level of participation in leisure activities and risk of aMCI in a prospective cohort of 437 community-residing subjects older than 75 years, initially free of dementia or aMCI, using Cox analysis adjusted for age, sex, education, and chronic illnesses. The authors derived Cognitive and Physical Activity Scales based on frequency of participation in individual activities. RESULTS: Over a median follow-up of 5.6 years, 58 subjects had development of aMCI. A one-point increase on the Cognitive (hazard ratio [HR] 0.95, 95% CI 0.91 to 0.99) but not Physical Activities Scale (HR 0.97, 95% CI 0.93 to 1.01) was associated with lower risk of aMCI. Subjects with Cognitive Activity scores in the highest (HR 0.46, 95% CI 0.24 to 0.91) and middle thirds (HR 0.52, 95% CI 0.29 to 0.96) had a lower risk of aMCI compared with subjects in the lowest third. The association persisted even after excluding subjects who converted to dementia within 2 years of meeting criteria for aMCI. CONCLUSIONS: Cognitive activity participation is associated with lower risk of development of amnestic mild cognitive impairment, even after excluding individuals at early stages of dementia. PMID- 16467496 TI - P38 MAP kinase mediates transforming growth factor-beta2 transcription in human keloid fibroblasts. AB - Keloids are abnormal fibrous growths of the dermis that develop only in response to wounding and represent a form of benign skin tumor. Previous studies have shown increased protein levels of TGF-beta in keloid tissue, suggesting a strong association with keloid formation leading us to examine mechanisms for why it is more highly expressed in keloids. Here, we use serum stimulation as an in vitro model to mimic a component of the wound microenvironment and examine differential gene expression in keloid human fibroblasts (KFs) vs. normal human fibroblasts (NFs). Transcription of TGF-beta2 was rapid and peaked between 1 and 6 h after serum stimulation in KFs vs. NFs. We confirmed increased TGF-beta activity in the conditioned medium from KFs, but not NFs. Inhibition of second messenger signaling pathways demonstrated that only the p38 MAPK inhibitor SB-203580 could block upregulation of TGF-beta2 following serum stimulation in KFs. Immunoblotting demonstrated that p38 MAPK was phosphorylated within 15 min and was maintained at a high level in KFs but not in NFs. The transcription factors activating transcription factor-2 and Elk-1 are activated by p38 MAPK, and also showed rapid and prolonged phosphorylation kinetics in KFs but not in NFs. In conclusion, increased TGF-beta2 transcription in response to serum stimulation in KFs appears to be mediated by the p38 MAPK pathway. This suggests the mechanism of keloid pathogenesis may be due in part to an inherent difference in how the fibroblasts respond to wounding. PMID- 16467497 TI - Dietary salt and hypertension: new molecular targets add more spice. PMID- 16467498 TI - How does salt retention raise blood pressure? AB - A critical question in hypertension research is: How is long-term blood pressure controlled? Excessive NaCl ingestion or NaCl retention by the kidneys and the consequent tendency toward plasma volume expansion lead to hypertension. Nevertheless, the precise mechanisms linking salt to high blood pressure are unresolved. The discovery of endogenous ouabain, an adrenocortical hormone, provided an important clue. Ouabain, a selective Na+ pump inhibitor, has cardiotonic and vasotonic effects. Plasma endogenous ouabain levels are significantly elevated in approximately 40% of patients with essential hypertension and in animals with several forms of salt-dependent hypertension. Also, prolonged ouabain administration induces hypertension in rodents. Mice with mutant Na+ pumps or Na/Ca exchangers (NCX) and studies with a ouabain antagonist and an NCX blocker are revealing the missing molecular mechanisms. These data demonstrate that alpha2 Na+ pumps and NCX1 participate in long-term regulation of vascular tone and blood pressure. Pharmacological agents or mutations in the alpha2 Na+ pump that interfere with the action of ouabain on the pump, and reduced NCX1 expression or agents that block NCX all impede the development of salt-dependent or ouabain-induced hypertension. Conversely, nanomolar ouabain, reduced alpha2 Na+ pump expression, and smooth muscle-specific overexpression of NCX1 all induce hypertension. Furthermore, ouabain and reduced alpha2 Na+ pump expression increase myogenic tone in isolated mesenteric small arteries in vitro, thereby tying these effects directly to the elevation of blood pressure. Thus, endogenous ouabain, and vascular alpha2 Na+ pumps and NCX1, are critical links between salt and hypertension. New pharmacological agents that act on these molecular links have potential in the clinical management of hypertension. PMID- 16467499 TI - Physiological role of the alpha1- and alpha2-isoforms of the Na+-K+-ATPase and biological significance of their cardiac glycoside binding site. AB - An interesting feature of Na+-K+-ATPase is that it contains four isoforms of the catalytic alpha-subunit, each with a tissue-specific distribution. Our laboratory has used gene targeting to define the functional role of the alpha1- and alpha2 isoforms. While knockout mice demonstrated the importance of the alpha1- and alpha2-isoforms for survival, the knockin mice, in which each isoform can be individually inhibited by ouabain and its function determined, demonstrated that both isoforms are regulators of cardiac muscle contractility. Another intriguing aspect of the Na+-K+-ATPase is that it contains a binding site for cardiac glycosides, such as digoxin. Conservation of this site suggests that it may have an in vivo role and that a natural ligand must exist to interact with this site. In fact, cardiac glycoside-like compounds have been observed in mammals. Our recent study demonstrates that the cardiac glycoside binding site of the Na+-K+ ATPase plays a role in the regulation of blood pressure and that it mediates both ouabain-induced and ACTH-induced hypertension in mice. Whereas chronic administration of ouabain or ACTH caused hypertension in wild-type mice, it had no effect on blood pressure in mice with a ouabain-resistant alpha2-isoform of Na+-K+-ATPase. Interestingly, animals with the ouabain-sensitive alpha1-isoform and a ouabain-resistant alpha2-isoform develop ACTH-induced hypertension to a greater extent than wild-type animals. Taken together, these results demonstrate that the cardiac glycoside binding of the Na+-K+-ATPase has a physiological role and suggests a function for a naturally occurring ligand that is stimulated by administration of ACTH. PMID- 16467500 TI - Rostafuroxin: an ouabain antagonist that corrects renal and vascular Na+-K+- ATPase alterations in ouabain and adducin-dependent hypertension. AB - The genetic and environmental heterogeneity of essential hypertension is responsible for the individual variability of antihypertensive therapy. An understanding of the molecular mechanisms underlying hypertension and related organ complications is a key aspect for developing new, effective, and safe antihypertensive agents able to cure the cause of the disease. Two mechanisms, among others, are involved in determining the abnormalities of tubular Na+ reabsorption observed in essential hypertension: the polymorphism of the cytoskeletal protein alpha-adducin and the increased circulating levels of endogenous ouabain (EO). Both lead to increased activity and expression of the renal Na+-K+ pump, the driving force for tubular Na transport. Morphological and functional vascular alterations have also been associated with EO. Rostafuroxin (PST 2238) is a new oral antihypertensive agent able to selectively antagonize EO, adducin pressor, and molecular effects. It is endowed with high potency and efficacy in reducing blood pressure and preventing organ hypertrophy in animal models representative of both adducin and EO mechanisms. At molecular level, in the kidney, Rostafuroxin antagonizes EO triggering of the Src-epidermal growth factor receptor (EGFr)-dependent signaling pathway leading to renal Na+-K+ pump, and ERK tyrosine phosphorylation and activation. In the vasculature, it normalizes the increased myogenic tone caused by nanomolar ouabain. A very high safety ratio and an absence of interaction with other mechanisms involved in blood pressure regulation, together with initial evidence of high tolerability and efficacy in hypertensive patients, indicate Rostafuroxin as the first example of a new class of antihypertensive agents designed to antagonize adducin and EO hypertensive mechanisms. PMID- 16467501 TI - Vascular Na+/Ca2+ exchanger: implications for the pathogenesis and therapy of salt-dependent hypertension. AB - The Na+/Ca2+ exchanger is an ion transporter that exchanges Na+ and Ca2+ in either Ca2+ efflux or Ca2+ influx mode, depending on membrane potential and transmembrane ion gradients. In arterial smooth muscle cells, the Na+/Ca2+ exchanger is thought to participate in the maintenance of vascular tone by regulating cytosolic Ca2+ concentration. Recent pharmacological and genetic engineering studies have revealed that the Ca2+ influx mode of vascular Na+/Ca2+ exchanger type-1 (NCX1) is involved in the pathogenesis of salt-dependent hypertension. SEA0400, a specific Na+/Ca2+ exchange inhibitor that preferentially blocks the Ca2+ influx mode, lowers arterial blood pressure in salt-dependent hypertensive models, but not in normotensive rats or other types of hypertensive rats. Furthermore, heterozygous mice with reduced expression of NCX1 are resistant to development of salt-dependent hypertension, whereas transgenic mice with vascular smooth muscle-specific overexpression of NCX1 readily develop hypertension after high-salt loading. SEA0400 reverses the cytosolic Ca2+ elevation and vasoconstriction induced by nanomolar ouabain, as well as humoral factors in salt-loaded animals. One possibility is that circulating endogenous cardiotonic steroids may be necessary for NCX1-mediated hypertension. These findings help to explain how arterial smooth muscle cells in blood vessels contribute to salt-elicited blood pressure elevation and suggest that NCX1 inhibitors might be therapeutically useful for salt-dependent hypertension. PMID- 16467502 TI - Role of potassium in regulating blood flow and blood pressure. AB - Unlike sodium, potassium is vasoactive; for example, when infused into the arterial supply of a vascular bed, blood flow increases. The vasodilation results from hyperpolarization of the vascular smooth muscle cell subsequent to potassium stimulation by the ion of the electrogenic Na+-K+ pump and/or activating the inwardly rectifying Kir channels. In the case of skeletal muscle and brain, the increased flow sustains the augmented metabolic needs of the tissues. Potassium ions are also released by the endothelial cells in response to neurohumoral mediators and physical forces (such as shear stress) and contribute to the endothelium-dependent relaxations, being a component of endothelium-derived hyperpolarization factor-mediated responses. Dietary supplementation of potassium can lower blood pressure in normal and some hypertensive patients. Again, in contrast to NaCl restriction, the response to potassium supplementation is slow to appear, taking approximately 4 wk. Such supplementation reduces the need for antihypertensive medication. "Salt-sensitive" hypertension responds particularly well, perhaps, in part, because supplementation with potassium increases the urinary excretion of sodium chloride. Potassium supplementation may even reduce organ system complications (e.g., stroke). PMID- 16467503 TI - Salt intake and depletion increase circulating levels of endogenous ouabain in normal men. AB - High-salt diets elevate circulating Na+ pump inhibitors, vascular resistance, and blood pressure. Ouabain induces a form of hypertension mediated via the alpha2 Na+ pump isoform and the calcium influx mode of the vascular sodium calcium exchanger (NCX). Whereas elevated levels of an endogenous ouabain (EO) and NCX have been implicated in salt-sensitive hypertension, acute changes in sodium balance do not affect plasma EO. This study investigated the impact of longer term alterations in sodium balance on the circulating levels and renal clearance of EO in normal humans. Thirteen normal men consumed a normal diet, high-salt diet, and hydrochlorothiazide (HCTZ), each for 5-day periods to alter sodium balance. EO and other humoral and urinary variables were determined daily. On a normal diet, urinary sodium excretion (140 +/- 16 meq/day), plasma EO (0.43 +/- 0.08 nmol/l) and urinary EO excretion (1.04 +/- 0.13 nmol/day) were at steady state. On the 3rd day of a high-salt diet, urine sodium excretion (315 +/- 28 meq/day), plasma EO (5.8 +/- 2.2 nmol/l), and the urinary EO excretion (1.69 +/- 0.27 nmol/day) were significantly increased, while plasma renin activity and aldosterone levels were suppressed. The salt-evoked increase in plasma EO was greater in older individuals, in subjects whose baseline circulating EO was higher, and in those with low renal clearance. During HCTZ, body weight decreased and plasma renin activity, aldosterone, and EO (1.71 +/- 0.77 nmol/l) rose, while urinary EO excretion remained within the normal range (1.44 +/- 0.31 nmol/day). Blood pressure fell in one subject during HCTZ. HPLC of the plasma extracts showed one primary peak of EO immunoreactivity with a retention time equivalent to ouabain. High-salt diets and HCTZ raise plasma EO by stimulating EO secretion, and a J-shaped curve relates sodium balance and EO in healthy men. Under normal dietary conditions, approximately 98% of the filtered load of EO is reabsorbed by the kidney, and differences in the circulating levels of EO are strongly influenced by secretion and urinary excretion of EO. The dramatic impact of high salt diets on plasma EO is consistent with its proposed role as a humoral vasoconstrictor that links salt intake with vascular function in hypertension. PMID- 16467504 TI - Cosignaling of adenosine and adenosine triphosphate in hypobaric hypoxia-induced hypothermia. AB - Purines, that is, adenosine and ATP, are not only products of metabolism but are also neurotransmitters. Indeed, purinergic neurotransmission is involved in thermoregulatory processes that occur during normoxia. Exposure to severe hypoxia elicits a sharp decrease in body core temperature (T(CO)), and adenosinergic mechanisms have been suspected to be responsible for this hypothermia. Because ATP per se and its metabolite adenosine could have complex interactions in some neural networks, we hypothesize that both adenosine and ATP are involved in the central mechanism of hypoxia-induced hypothermia. Their role in the thermoregulatory process was therefore investigated in a 24-h hypobaric hypoxia (Fi(O2) = 10%), using CGS-15943, a nonselective antagonist of adenosine receptors, and suramin, an ATP receptor antagonist. T(CO) and spontaneous activity (A(S)) were monitored by telemetry in conscious rats, receiving CGS 15943 (10 mg/kg ip), suramin (7 nmol icv), or both. The same treatments were done in normoxia to evaluate the specificity of their thermoregulatory action observed in hypoxia. Suramin/CGS-15943 treatment blunted the profound hypothermia observed in control rats throughout the hypoxia exposure, whereas CGS-15943 treatment blunted hypothermia during only 3 h, and suramin treatment had no effect. These results suggest that suramin potentiates the CGS-15943 effects and consequently that adenosine and ATP signaling act in synergy. In normoxia, suramin/CGS-15943 induced an increase in T(CO) but to a far lesser extent than observed in hypoxia. Thus it might be suggested that the suramin/CGS-15943 blunting of hypoxia-induced hypothermia would be specific to hypoxia-induced mechanisms. PMID- 16467505 TI - Effect of apocynin treatment on renal expression of COX-2, NOS1, and renin in Wistar-Kyoto and spontaneously hypertensive rats. AB - Macula densa (MD) cells of the juxtaglomerular apparatus (JGA) synthesize type 1 nitric oxide synthase (NOS1) and type 2 cyclooxygenase (COX-2). Both nitric oxide (NO) and prostaglandins have been considered to mediate or modulate the control of renin secretion. Reactive oxygen species (ROS) produced locally by NADPH oxidase may influence NO bioavailability. We have tested the hypothesis that in hypertension elevated ROS levels may modify the expression of NOS1 and COX-2 in the JGA, thereby interacting with juxtaglomerular signaling. To this end, spontaneously hypertensive rats (SHR) and Wistar-Kyoto control rats (WKY) received the specific NADPH oxidase inhibitor, apocynin, during 3 wk. Renal functional and histochemical parameters, plasma renin activity (PRA), and as a measure of ROS activity, urinary isoprostane excretion (IP) were evaluated. Compared with WKY, IP levels in untreated SHR were 2.2-fold increased, and NOS1 immunoreactiviy (IR) of JGA 1.5-fold increased, whereas COX-2 IR was reduced to 35%, renin IR to 51%, and PRA to 7%. Apocynin treatment reduced IP levels in SHR to 52%, NOS1 IR to 69%, and renin IR to 62% of untreated SHR, whereas renin mRNA, COX-2 IR, glomerular filtration rate, PRA, and systolic blood pressure remained unchanged. WKY revealed no changes under apocynin treatment. These data show that NADPH oxidase is an important contributor to elevated levels of ROS in hypertension. Upregulation of MD NOS1 in SHR may have the potential of blunting the functional impact of ROS at the level of bioavailable NO. Downregulated COX-2 and renin levels in SHR are apparently unrelated to oxidative stress, since apocynin treatment had no effect on these parameters. PMID- 16467506 TI - Sprinting and endurance for cyclists and runners. PMID- 16467507 TI - Estrogen increases protective proteins following trauma and hemorrhage. PMID- 16467508 TI - Rosiglitazone (Avandia) and macular edema. PMID- 16467509 TI - Kinases bicker over an ion channel. PMID- 16467510 TI - What can immediate-early gene expression tell us about spatial memory retrieval? PMID- 16467511 TI - From invertebrate olfaction to human cognition: emerging computational functions of synchronized oscillatory activity. PMID- 16467512 TI - Olfactory computations and network oscillation. PMID- 16467513 TI - The cognitive correlates of human brain oscillations. PMID- 16467514 TI - Network oscillations: emerging computational principles. PMID- 16467515 TI - Single Ih channels in pyramidal neuron dendrites: properties, distribution, and impact on action potential output. AB - The hyperpolarization-activated cation current (Ih) plays an important role in regulating neuronal excitability, yet its native single-channel properties in the brain are essentially unknown. Here we use variance-mean analysis to study the properties of single Ih channels in the apical dendrites of cortical layer 5 pyramidal neurons in vitro. In these neurons, we find that Ih channels have an average unitary conductance of 680 +/- 30 fS (n = 18). Spectral analysis of simulated and native Ih channels showed that there is little or no channel flicker below 5 kHz. In contrast to the uniformly distributed single-channel conductance, Ih channel number increases exponentially with distance, reaching densities as high as approximately 550 channels/microm2 at distal dendritic sites. These high channel densities generate significant membrane voltage noise. By incorporating a stochastic model of Ih single-channel gating into a morphologically realistic model of a layer 5 neuron, we show that this channel noise is higher in distal dendritic compartments and increased threefold with a 10-fold increased single-channel conductance (6.8 pS) but constant Ih current density. In addition, we demonstrate that voltage fluctuations attributable to stochastic Ih channel gating impact on action potential output, with greater spike-timing precision in models with the experimentally determined single channel conductance. These data suggest that, in the face of high current densities, the small single-channel conductance of Ih is critical for maintaining the fidelity of action potential output. PMID- 16467516 TI - MTH1, an oxidized purine nucleoside triphosphatase, suppresses the accumulation of oxidative damage of nucleic acids in the hippocampal microglia during kainate induced excitotoxicity. AB - Enhanced oxidative stress has been implicated in the excitotoxicity of the CNS, and 8-oxo-7,8-dihydro-guanine (8-oxoG), a major type of oxidative damage in nucleic acids, was reported to be accumulated in the rat hippocampus after kainate administration. We herein showed that the 8-oxoG levels in mitochondrial DNA and cellular RNA increased significantly in the CA3 subregion of the mouse hippocampus 6-12 h after kainate administration but returned to basal levels within a few days. Laser-scanning confocal microscopy revealed the 8-oxoG accumulation in mitochondrial DNA to be remarkable in CA3 microglia, whereas that in nuclear DNA or cellular RNA was also detected in the CA3 pyramidal cells and astrocytes. 8-oxoG accumulation in cellular DNA or RNA should be suppressed by MutT homolog 1 (MTH1) with 8-oxo-dGTPase (8-oxo-7,8-dihydro-2'-deoxyguanosine triphosphatase) activity and 8-oxoG-DNA glycosylase 1 (OGG1) with 8-oxoG DNA glycosylase activity. We thus examined the expression level of MTH1 and OGG1 in the mouse hippocampus after kainate administration. The Mth1 mRNA level decreased soon after kainate administration and then quickly recovered beyond the basal level, and a continuously increased MTH1 protein level was observed, whereas the Ogg1 mRNA level remained constant. MTH1-null and wild-type mice exhibited a similar degree of CA3 neuron loss after kainate administration; however, the 8 oxoG levels that accumulated in mitochondrial DNA and cellular RNA in the CA3 microglia significantly increased in the MTH1-null mice in comparison with wild type mice, thus demonstrating that MTH1 efficiently suppresses the accumulation of 8-oxoG in both cellular DNA and RNA in the hippocampus, especially in microglia, caused by excitotoxicity. PMID- 16467517 TI - Preoptic glutamate facilitates male sexual behavior. AB - The medial preoptic area (MPOA) is a critical regulatory site for the control of male sexual behavior. We first measured glutamate in 2 min microdialysate samples from the MPOA before, during, and after copulation by male rats. There was a slight [approximately 140% of baseline (BL)] rise in extracellular glutamate when the female was presented, a significant increase (approximately 170% of BL) during periods of mounting and intromitting, and a very large increase in samples collected during ejaculation (approximately 300% of BL). A precipitous fall in levels occurred in the first postejaculatory sample; the magnitude of this fall was highly correlated with the length of the postejaculatory interval of quiescence. In experiment 2, we reverse-dialyzed a mixture of glutamate uptake inhibitors into the MPOA before and during mating; control animals received artificial CSF. The mixture increased extracellular glutamate (approximately 280% of BL), increased the number of ejaculations in the 40 min test, decreased ejaculation latency, and decreased the postejaculatory latency to resume copulation. These data, together with other findings that glutamate in the MPOA can elicit genital reflexes in anesthetized rats and that glutamate receptor antagonists in the MPOA impair copulation, strongly suggest that MPOA glutamate is a major facilitator of copulation and that the postejaculatory fall in glutamate regulates the postejaculatory interval. PMID- 16467518 TI - Steering by hearing: a bat's acoustic gaze is linked to its flight motor output by a delayed, adaptive linear law. AB - Adaptive behaviors require sensorimotor computations that convert information represented initially in sensory coordinates to commands for action in motor coordinates. Fundamental to these computations is the relationship between the region of the environment sensed by the animal (gaze) and the animal's locomotor plan. Studies of visually guided animals have revealed an anticipatory relationship between gaze direction and the locomotor plan during target-directed locomotion. Here, we study an acoustically guided animal, an echolocating bat, and relate acoustic gaze (direction of the sonar beam) to flight planning as the bat searches for and intercepts insect prey. We show differences in the relationship between gaze and locomotion as the bat progresses through different phases of insect pursuit. We define acoustic gaze angle, theta(gaze), to be the angle between the sonar beam axis and the bat's flight path. We show that there is a strong linear linkage between acoustic gaze angle at time t [theta(gaze)(t)] and flight turn rate at time t + tau into the future [theta(flight) (t + tau)], which can be expressed by the formula theta(flight) (t + tau) = ktheta(gaze)(t). The gain, k, of this linkage depends on the bat's behavioral state, which is indexed by its sonar pulse rate. For high pulse rates, associated with insect attacking behavior, k is twice as high compared with low pulse rates, associated with searching behavior. We suggest that this adjustable linkage between acoustic gaze and motor output in a flying echolocating bat simplifies the transformation of auditory information to flight motor commands. PMID- 16467519 TI - Presynaptic terminals independently regulate synaptic clustering and autophagy of GABAA receptors in Caenorhabditis elegans. AB - Synaptic clustering of GABAA receptors is important for the function of inhibitory synapses, influencing synapse strength and, consequently, the balance of excitation and inhibition in the brain. Presynaptic terminals are known to induce GABAA receptor clustering during synaptogenesis, but the mechanisms of cluster formation and maintenance are not known. To study how presynaptic neurons direct the formation of GABAA receptor clusters, we have investigated GABAA receptor localization in postsynaptic cells that fail to receive presynaptic contacts in Caenorhabditis elegans. Postsynaptic muscles in C. elegans receive acetylcholine and GABA motor innervation, and GABAA receptors cluster opposite GABA terminals. Selective loss of GABA inputs caused GABAA receptors to be diffusely distributed at or near the muscle cell surface, confirming that GABA presynaptic terminals induce GABAA receptor clustering. In contrast, selective loss of acetylcholine innervation had no effect on GABAA receptor localization. However, loss of both GABA and acetylcholine inputs together caused GABAA receptors to traffic to intracellular autophagosomes. Autophagosomes normally transport bulk cytoplasm to the lysosome for degradation. However, we show that GABAA receptors traffic to autophagosomes after endocytic removal from the cell surface and that acetylcholine receptors in the same cells do not traffic to autophagosomes. Thus, autophagy can degrade cell-surface receptors and can do so selectively. Our results show that presynaptic terminals induce GABAA receptor clustering by independently controlling synaptic localization and surface stability of GABAA receptors. They also demonstrate a novel function for autophagy in GABAA receptor degradative trafficking. PMID- 16467520 TI - Distinct roles for Ras-guanine nucleotide-releasing factor 1 (Ras-GRF1) and Ras GRF2 in the induction of long-term potentiation and long-term depression. AB - NMDA-type glutamate receptors (NMDARs) contribute to many forms of long-term potentiation (LTP) and long-term depression (LTD). NMDARs are heteromers containing calcium-permeating neuronal receptor 1 (NR1) subunits and a variety of NR2 subunits. Evidence suggests that, in the CA1 region of the hippocampus, NR2A containing NMDARs promote LTP whereas NR2B-containing receptors promote LTD. However, the calcium sensors that distinguish between these signals to promote the appropriate form of synaptic plasticity are not known. Ras-guanine nucleotide releasing factor 1 (Ras-GRF1) and Ras-GRF2 are highly similar calcium-stimulated exchange factors that activate Ras and Rac GTPases. Here, using a set of Ras-GRF knock-out mice, we show that Ras-GRF2 contributes predominantly to the induction of NMDAR-dependent LTP, whereas Ras-GRF1 contributes predominantly to the induction of NMDAR-dependent LTD in the CA1 region of the hippocampus of postpubescent mice (postnatal days 25-36). In contrast, neither Ras-GRF protein influences synaptic plasticity in prepubescent mice (postnatal days 14-18). Ras GRF2 mediates signaling from (R)-[(S)-1-(4-bromo-phenyl)-ethylamino]-(2,3-dioxo 1,2,3,4-tetrahydroquinoxalin-5-yl)-methyl-phosphonic acid-sensitive (NVP-AAM077 sensitive) (NR2A-containing) NMDARs to the Ras effector extracellular signal related protein kinase 1/2 (Erk1/2) mitogen-activated protein (MAP) kinase, a promoter of NMDAR-induced LTP at this site. In contrast, Ras-GRF1 mediates signaling from ifenprodil-sensitive (NR2B-containing) NMDARs to the Rac effector p38 MAP kinase, a promoter of LTD. These findings show that, despite their similar functional domain organization, Ras-GRF1 and Ras-GRF2 mediate opposing forms of synaptic plasticity by coupling different classes of NMDARs to distinct MAP kinase pathways. Moreover, the postnatal appearance of Ras-GRF-dependent LTP and LTD coincides with the emergence of hippocampal-dependent behavior, implying that Ras-GRF proteins contribute to forms of synaptic plasticity that are required specifically for mature hippocampal function. PMID- 16467521 TI - A novel role for Sema3A in neuroprotection from injury mediated by activated microglia. AB - Microglia exist under physiological conditions in a resting state but become activated after neuronal injury. Recent studies have highlighted the reciprocal role of neurons in controlling both the number and activity of microglia. In this study, microglia derived from newborn rat cortices were cultured and activated by interferon-gamma (IFNgamma) treatment, then exposed to recombinant Sema3A or conditioned medium derived from stressed embryonic cortical neurons. We found that activation of microglia by IFNgamma induced differential upregulation of the semaphorin receptors Plexin-A1 and Neuropilin-1. This result was confirmed by Northern blotting, reverse transcription-PCR, and Western blotting. Furthermore, recombinant Sema3A induced apoptosis of microglia when added to the in vitro culture, and a similar result was obtained on activated microglia when Sema3A was produced by stressed neurons. Using an in vivo model of microglia activation by striatal injection of lipopolysaccharide demonstrated a corresponding upregulation of Plexin-A1 and Neuropilin-1 in activated microglia and enhanced production of Sema3A by stressed adult neurons. These results suggest a novel semaphorin-mediated mechanism of neuroprotection whereby stressed neurons can protect themselves from further damage by activated microglia. PMID- 16467522 TI - Block of inferior olive gap junctional coupling decreases Purkinje cell complex spike synchrony and rhythmicity. AB - Inferior olivary (IO) neurons are electrotonically coupled by gap junctions. This coupling is thought to underlie synchronous complex spike (CS) activity generated by the olivocerebellar system in Purkinje cells, and also has been hypothesized to be necessary for IO neurons to generate spontaneous oscillatory activity. These characteristics of olivocerebellar activity have been proposed to be central to the role of this system in motor coordination. However, the relationship of gap junction coupling between IO neurons to synchronous and rhythmic CS activity has never been directly tested. Thus, to address this issue, multiple electrode recordings were obtained from crus 2a Purkinje cells, and carbenoxolone, a gap junction blocker, was injected into the IO. Carbenoxolone reduced CS synchrony by 50% overall, but in some experiments, >80% reductions were achieved. Carbenoxolone also reduced the average firing rate by 50%, suggesting that electrical coupling is a significant source of excitation for IO neurons. Moreover, carbenoxolone caused a reduction in the approximately 10 Hz rhythmicity of CS activity, and this reduction was correlated with the extent to which the injection reduced CS synchrony. Lastly, carbenoxolone was found to reverse or prevent changes in synchrony that are normally induced by injection of GABAA and glutamate receptor antagonists into the IO, suggesting that the effects of these drugs on CS synchrony patterns require electrical coupling of IO neurons. In sum, our results provide direct evidence that electrical coupling of IO neurons underlies synchronous CS activity, and suggest important roles for this coupling in shaping other aspects of IO spiking patterns. PMID- 16467523 TI - Chronic intermittent ethanol-induced switch of ethanol actions from extrasynaptic to synaptic hippocampal GABAA receptors. AB - Alcohol withdrawal syndrome (AWS) symptoms include hyperexcitability, anxiety, and sleep disorders. Chronic intermittent ethanol (CIE) treatment of rats with subsequent withdrawal of ethanol (EtOH) reproduced AWS symptoms in behavioral assays, which included tolerance to the sleep-inducing effect of acute EtOH and its maintained anxiolytic effect. Electrophysiological assays demonstrated a CIE induced long-term loss of extrasynaptic GABAA receptor (GABAAR) responsiveness and a gain of synaptic GABAAR responsiveness of CA1 pyramidal and dentate granule neurons to EtOH that we were able to relate to behavioral effects. After CIE treatment, the alpha4 subunit-preferring GABAAR ligands 4,5,6,7 tetrahydroisoxazolo[5,4-c]pyridin-3-ol, La3+, and Ro15-4513 (ethyl-8-azido-5,6 dihydro-5-methyl-6-oxo-4H-imidazo[1,5alpha][1,4]benzodiazepine-3-carboxylate) exerted decreased effects on extrasynaptic currents but had increased effects on synaptic currents. Electron microscopy revealed an increase in central synaptic localization of alpha4 but not delta subunits within GABAergic synapses on the dentate granule cells of CIE rats. Recordings in dentate granule cells from delta subunit-deficient mice revealed that this subunit is not required for synaptic GABAAR sensitivity to low [EtOH]. The profound alterations in EtOH sensitivity and alpha4 subunit localization at hippocampal GABAARs of CIE rats suggest that such changes in these and other relevant brain circuits may contribute to the development of tolerance to the sleep-inducing effects and long-term dependence on alcohol. PMID- 16467524 TI - Synaptic transmission mediated by internal calcium stores in rod photoreceptors. AB - Retinal rod photoreceptors are depolarized in darkness to approximately -40 mV, a state in which they maintain sustained glutamate release despite low levels of calcium channel activation. Blocking voltage-gated calcium channels or ryanodine receptors (RyRs) at the rod presynaptic terminal suppressed synaptic communication to bipolar cells. Spontaneous synaptic events were also inhibited when either of these pathways was blocked. This indicates that both calcium influx and calcium release from internal stores are required for the normal release of transmitter of the rod. RyR-independent release can be evoked by depolarization of a rod to a supraphysiological potential (-20 mV) that activates a large fraction of voltage-gated channels. However, this calcium channel mediated release depletes rapidly if RyRs are blocked, indicating that RyRs support prolonged glutamate release. Thus, the rod synapse couples a small transmembrane calcium influx with a RyR-dependent amplification mechanism to support continuous vesicle release. PMID- 16467525 TI - Impaired neuronal positioning and dendritogenesis in the neocortex after cell autonomous Dab1 suppression. AB - Reelin and Disabled 1 (Dab1) are essential for positioning migrating neurons in the developing neocortex. Cell-autonomous RNA interference-mediated suppression of Dab1 in migrating neurons destined for layer 2/3 shifted the median position of these cells to deeper positions within the cortex. At the time of migration arrest [embryonic day 20 (E20) to E21], Dab1-suppressed cells were underrepresented in the upper approximately 40 microm of the cortex compared with controls, suggesting that Dab1 is essential for somal translocation through the cell-dense cortical plate. Closer examination of the morphology of Dab1 suppressed neurons at E20 revealed simplified leading processes that are less likely to contact the marginal zone (MZ), in which high levels of Reelin are expressed. Examination of Dab1-suppressed cells 3 d later (postnatal day 2) revealed simplified dendrites that are also less likely to contact the MZ. These data reveal a cell-autonomous role of Dab1 in dendritogenesis in the neocortex and suggest that remodeling of the leading process of a migrating neuron into a nascent dendrite by Reelin/Dab1 signaling plays an important role in cell positioning. PMID- 16467526 TI - Telencephalin slows spine maturation. AB - Dendritic filopodia are highly dynamic structures, and morphological maturation from dendritic filopodia to spines is intimately associated with the stabilization and strengthening of synapses during development. Here, we report that telencephalin (TLCN), a cell adhesion molecule belonging to the Ig superfamily, is a negative regulator of spine maturation. Using cultured hippocampal neurons, we examined detailed localization and functions of TLCN in spine development and synaptogenesis. At early stages of synaptogenesis, TLCN immunoreactivity gradually increased and was present in dendritic shafts and filopodia. At later stages, TLCN tended to be excluded from mature spine synapses in which PSD-95 (postsynaptic density-95) clusters were apposed to presynaptic synaptophysin clusters. To elucidate the function of TLCN in spine maturation, we analyzed the dendrite morphology of TLCN-overexpressing and TLCN-deficient neurons. Overexpression of TLCN caused a dramatic increase in the density of dendritic filopodia and a concomitant decrease in the density of spines. Conversely, TLCN-deficient mice showed a decreased density of filopodia and an acceleration of spine maturation in vitro as well as in vivo. These results demonstrate that TLCN normally slows spine maturation by promoting the filopodia formation and negatively regulating the filopodia-to-spine transition. In addition, we found that spine heads of mature neurons were wider in TLCN deficient mice compared with wild-type mice. Thus, the preservation of immature synapses by TLCN may be an essential step for refinement of functional neural circuits in the telencephalon, that take charge of higher brain functions such as learning, memory, and emotion. PMID- 16467527 TI - Direct GABAergic and glycinergic inhibition of the substantia gelatinosa from the rostral ventromedial medulla revealed by in vivo patch-clamp analysis in rats. AB - Stimulation of the rostral ventromedial medulla (RVM) is believed to exert analgesic effects through the activation of the serotonergic system descending to the spinal dorsal horn; however, how nociceptive transmission is modulated by the descending system has not been fully clarified. To investigate the inhibitory mechanisms affected by the RVM, an in vivo patch-clamp technique was used to record IPSCs from the substantia gelatinosa (SG) of the spinal cord evoked by chemical (glutamate injection) and electrical stimulation (ES) of the RVM in adult rats. In the voltage-clamp mode, the RVM glutamate injection and RVM-ES produced an increase in both the frequency and amplitude of IPSCs in SG neurons that was not blocked by glutamate receptor antagonists. Serotonin receptor antagonists were unexpectedly without effect, but a GABAA receptor antagonist, bicuculline, or a glycine receptor antagonist, strychnine, completely suppressed the RVM stimulation-induced increase in IPSCs. The RVM-ES-evoked IPSCs showed fixed latency and no failure at 20 Hz stimuli with a conduction velocity of >3 m/s (3.1-20.7 m/s), suggesting descending monosynaptic GABAergic and/or glycinergic inputs from the RVM to the SG through myelinated fibers. In the current-clamp mode, action potentials elicited by noxious mechanical stimuli applied to the receptive field of the ipsilateral hindlimb were suppressed by the RVM-ES in more than half of the neurons tested (63%; 10 of 16). These findings suggest that the RVM-mediated antinociceptive effects on noxious inputs to the SG may be exerted preferentially by the direct GABAergic and glycinergic pathways to the SG. PMID- 16467528 TI - Presynaptic I1-imidazoline receptors reduce GABAergic synaptic transmission in striatal medium spiny neurons. AB - Imidazoline receptors are expressed widely in the CNS. In the present study, whole-cell patch-clamp recordings were made from medium spiny neurons in dorsal striatum slices from the rat brain, and the roles of I1-imidazoline receptors in the modulation of synaptic transmission were studied. Moxonidine, an I1 imidazoline receptor agonist, decreased the GABAA receptor-mediated IPSCs in a concentration-dependent manner. However, glutamate-mediated EPSCs were hardly affected. The depression of IPSCs by moxonidine was antagonized by either idazoxan or efaroxan, which are both imidazoline receptor antagonists containing an imidazoline moiety. In contrast, yohimbine and SKF86466 (6-chloro-2,3,4,5 tetrahydro-3-methyl-1H-3-benzazepine), which are alpha2-adrenergic receptor antagonists with no affinity for imidazoline receptors, did not affect the moxonidine-induced inhibition of IPSCs. Moxonidine increased the paired-pulse ratio and reduced the frequency of miniature IPSCs without affecting their amplitude, indicating that this agent inhibits IPSCs via presynaptic mechanisms. Moreover, the sulfhydryl alkylating agent N-ethylmaleimide (NEM) significantly reduced the moxonidine-induced inhibition of IPSCs. Thus, the activation of presynaptic I1-imidazoline receptors decreases GABA-mediated inhibition of medium spiny neurons in the striatum, in which NEM-sensitive proteins such as G(i/o) type G-proteins play an essential role. The adenylate cyclase activator forskolin partly opposed IPSC inhibition elicited by subsequently applied moxonidine. Furthermore, the protein kinase C (PKC) activator phorbol 12,13-dibutyrate attenuated and the PKC inhibitor chelerythrine potentiated the moxonidine-induced inhibition of IPSCs. These results suggest that IPSC inhibition via presynaptic I1-imidazoline receptors involves intracellular adenylate cyclase activity and is influenced by static PKC activity in the striatum. PMID- 16467530 TI - Integrins control dendritic spine plasticity in hippocampal neurons through NMDA receptor and Ca2+/calmodulin-dependent protein kinase II-mediated actin reorganization. AB - The formation of dendritic spines during development and their structural plasticity in the adult brain are critical aspects of synaptogenesis and synaptic plasticity. Many different factors and proteins have been shown to control dendritic spine development and remodeling (Ethell and Pasquale, 2005). The extracellular matrix (ECM) components and their cell surface receptors, integrins, have been found in the vicinity of synapses and shown to regulate synaptic efficacy and play an important role in long-term potentiation (Bahr et al., 1997; Chavis and Westbrook, 2001; Chan et al., 2003; Lin et al., 2003; Bernard-Trifilo et al., 2005). Although molecular mechanisms by which integrins affect synaptic efficacy have begun to emerge, their role in structural plasticity is poorly understood. Here, we show that integrins are involved in spine remodeling in cultured hippocampal neurons. The treatment of 14 d in vitro hippocampal neurons with arginine-glycine-aspartate (RGD)-containing peptide, an established integrin ligand, induced elongation of existing dendritic spines and promoted formation of new filopodia. These effects were also accompanied by integrin-dependent actin reorganization and synapse remodeling, which were partially inhibited by function-blocking antibodies against beta1 and beta3 integrins. This actin reorganization was blocked with the NMDA receptor (NMDAR) antagonist MK801 [(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10 imine hydrogen maleate]. The Ca2+/calmodulin-dependent protein kinase II (CaMKII) inhibitor KN93 (N-[2-[N-(4-chlorocinnamyl)-N-methylaminomethyl]phenyl]-N-(2 hydroxyethyl)-4-methoxybenzenesulfonamide) also suppressed RGD-induced actin reorganization and synapse remodeling. Our findings show that integrins control ECM-mediated spine remodeling in hippocampal neurons through NMDAR/CaMKII dependent actin reorganization. PMID- 16467529 TI - Molecular reconstruction of nodes of Ranvier after remyelination by transplanted olfactory ensheathing cells in the demyelinated spinal cord. AB - Myelin-forming glial cells transplanted into the demyelinated spinal cord can form compact myelin and improve conduction properties. However, little is known of the expression and organization of voltage-gated ion channels in the remyelinated central axons or whether the exogenous cells provide appropriate signaling for the maturation of nodes of Ranvier. Here, we transplanted olfactory ensheathing cells from green fluorescent protein (GFP)-expressing donor rats [GFP olfactory ensheathing cells (OECs)] into a region of spinal cord demyelination and found extensive remyelination, which included the development of mature nodal, paranodal, and juxtaparanodal domains, as assessed by ultrastructural, immunocytochemical, and electrophysiological analyses. In remyelinated axons, Nav1.6 was clustered at nodes, whereas Kv1.2 was aggregated in juxtaparanodal regions, recapitulating the distribution of these channels within mature nodes of uninjured axons. Moreover, the recruitment of Nav and Kv channels to specific membrane domains at remyelinated nodes persisted for at least 8 weeks after GFP OEC transplantation. In vivo electrophysiological recordings demonstrated enhanced conduction along the GFP-OEC-remyelinated axons. These findings indicate that, in addition to forming myelin, engrafted GFP-OECs provide an environment that supports the development and maturation of nodes of Ranvier and the restoration of impulse conduction in central demyelinated axons. PMID- 16467531 TI - Loss of leukemia inhibitory factor receptor beta or cardiotrophin-1 causes similar deficits in preganglionic sympathetic neurons and adrenal medulla. AB - Leukemia inhibitory factor (LIF) receptor beta (LIFRbeta) is a receptor for a variety of neurotrophic cytokines, including LIF, ciliary neurotrophic factor (CNTF), and cardiotrophin-1 (CT-1). These cytokines play an essential role for the survival and maintenance of developing and postnatal somatic motoneurons. CNTF may also serve the maintenance of autonomic, preganglionic sympathetic neurons (PSNs) in the spinal cord, as suggested by its capacity to prevent their death after destruction of one of their major targets, the adrenal medulla. Although somatic motoneurons and PSNs share a common embryonic origin, they are distinct in several respects, including responses to lesions. We have studied PSNs in mice with targeted deletions of the LIFRbeta or CT-1 genes, respectively. We show that LIF, CNTF, and CT-1 are synthesized in embryonic adrenal gland and spinal cord and that PSNs express LIFRbeta. In embryonic day 18.5 LIFRbeta (-/-) and CT-1 (-/-) mice, PSNs were reduced by approximately 20%. PSNs projecting to the adrenal medulla were more severely affected (-55%). Although LIFRbeta (-/-) mice revealed normal numbers of adrenal chromaffin cells and axons terminating on chromaffin cells, levels of adrenaline and numbers of adrenaline-synthesizing cells were significantly reduced. We conclude that activation of LIFRbeta is required for normal development of PSNs and one of their prominent targets, the adrenal medulla. Thus, both somatic motoneurons and PSNs in the spinal cord depend on LIFRbeta signaling for their development and maintenance, although PSNs seem to be overall less affected than somatic motoneurons by LIFRbeta deprivation. PMID- 16467532 TI - Disinhibition opens the gate to pathological pain signaling in superficial neurokinin 1 receptor-expressing neurons in rat spinal cord. AB - Blockade of local spinal cord inhibition mimics the behavioral hypersensitivity that manifests in chronic pain states. This suggests that there is a pathway capable of mediating allodynia/hyperalgesia that exists but is normally under strong inhibitory control. Lamina I and III neurokinin 1 (NK1) receptor expressing (NK1R+) dorsal horn neurons, many of which are projection neurons, are required for the development of this hypersensitivity and are therefore likely to be a component of this proposed pathway. To investigate, whole-cell patch-clamp recordings were made from lamina I and III NK1R+ neurons in the spinal cord slice preparation with attached dorsal root. Excitatory postsynaptic currents were recorded in response to electrical stimulation of the dorsal root. Lamina I NK1R+ neurons were shown to receive high-threshold (Adelta/C fiber) monosynaptic input, whereas lamina III NK1R+ neurons received low-threshold (Abeta fiber) monosynaptic input. In contrast, lamina I neurons lacking NK1 receptor (NK1R-) received polysynaptic A fiber input. Blockade of local GABAergic and glycinergic inhibition with bicuculline (10 microm) and strychnine (300 nm), respectively, revealed significant A fiber input to lamina I NK1R+ neurons that was predominantly Abeta fiber mediated. This novel A fiber input was polysynaptic in nature and required NMDA receptor activity to be functional. In lamina I NK1R- and lamina III NK1R+ neurons, disinhibition enhanced control-evoked responses, and this was also NMDA receptor dependent. These disinhibition-induced changes, in particular the novel polysynaptic low-threshold input onto lamina I NK1R+ neurons, may be an underlying component of the hypersensitivity present in chronic pain states. PMID- 16467533 TI - Small-conductance Ca2+-activated K+ channel type 2 (SK2) modulates hippocampal learning, memory, and synaptic plasticity. AB - Apamin-sensitive, small-conductance, Ca2+-activated K+ channels (SK channels) modulate neuronal excitability in CA1 neurons. Blocking all SK channel subtypes with apamin facilitates the induction of hippocampal synaptic plasticity and enhances hippocampal learning. In CA1 dendrites, SK channels are activated by Ca2+ through NMDA receptors and restrict glutamate-mediated EPSPs. Studies of SK channel knock-out mice reveal that of the three apamin-sensitive SK channel subunits (SK1-SK3), only SK2 subunits are necessary for the apamin-sensitive currents in CA1 hippocampal neurons. To determine the specific influence of SK2 channels on hippocampal synaptic plasticity, learning, and memory, we used gene targeting through homologous recombination in embryonic stem cells to generate transgenic mice that overexpress SK2 subunits by 10-fold (SK2+/T). In these mice, the apamin-sensitive current in CA1 neurons was increased by approximately fourfold, relative to wild-type (WT) littermates. In addition, the amplitude of synaptically evoked EPSPs recorded from SK2+/T CA1 neurons increased twice as much in response to SK channel blockade relative to EPSPs recorded from WT CA1 neurons. Consistent with this, SK2 overexpression reduced long-term potentiation after high-frequency stimulation compared with WT littermates and severely impaired learning in both hippocampus- and amygdala-dependent tasks. We conclude that SK2 channels regulate hippocampal synaptic plasticity and play a critical role in modulating mechanisms of learning and memory. PMID- 16467534 TI - Site of action potential initiation in layer 5 pyramidal neurons. AB - Fundamental to an understanding of how neurons integrate synaptic input is the knowledge of where within a neuron this information is converted into an output signal, the action potential. Although it has been known for some time that action potential initiation occurs within the axon of neurons, the precise location has remained elusive. Here, we provide direct evidence using voltage sensitive dyes that the site of action potential initiation in cortical layer 5 pyramidal neurons is approximately 35 microm from the axon hillock. This was the case during action potential generation under a variety of conditions, after axonal inhibition, and at different stages of development. Once initiated action potentials propagated down the axon in a saltatory manner. Experiments using local application of low-sodium solution and TTX, as well as an investigation of the influence of axonal length on action potential properties, provided evidence that the initial 40 microm of the axon is essential for action potential generation. To morphologically identify the relationship between the site of action potential initiation and axonal myelination, we labeled oligodendrocytes supplying processes to the proximal region of the axon. These experiments indicated that the axon initial segment was approximately 40 mcirom in length, and the first node of Ranvier was approximately 90 microm from the axon hillock. Experiments targeting the first node of Ranvier suggested it was not involved in action potential initiation. In conclusion, these results indicate that, in layer 5 pyramidal neurons, action potentials are generated in the distal region of the axon initial segment. PMID- 16467535 TI - Cell-specific repressor or enhancer activities of Deaf-1 at a serotonin 1A receptor gene polymorphism. AB - The serotonin-1A (5-HT1A) receptor is the primary somatodendritic autoreceptor that inhibits the activity of serotonergic raphe neurons and is also expressed in nonserotonergic cortical and limbic neurons. Alterations in 5-HT1A receptor levels are implicated in mood disorders, and a functional C(-1019)G 5-HT1A promoter polymorphism has been associated with depression, suicide, and panic disorder. We examined the cell-specific activity of identified transcription factors, human nuclear deformed epidermal autoregulatory factor-1 (DEAF-1) related (NUDR)/Deaf-1 and Hes5, at the 5-HT1A C(-1019) site. In serotonergic raphe RN46A cells, Deaf-1 and Hes5 repressed the 5-HT1A receptor gene at the C( 1019)-allele but not the G(-1019)-allele. However, in nonserotonergic cells that express 5-HT1A receptors (septal SN48, neuroblastoma SKN-SH, and neuroblastoma/glioma NG108-15 cells), Deaf-1 enhanced 5-HT1A promoter activity at the C(-1019)-allele but not the G-allele, whereas Hes5 repressed in all cell types. The enhancer activity of Deaf-1 was orientation independent and competed out Hes5 repression. To test whether Deaf-1 activity is intrinsic, the activity of a Gal4DBD (DNA binding domain)-Deaf-1 fusion protein at a heterologous Gal4 DNA element was examined. Gal4DBD-Deaf-1 repressed transcription in RN46A cells but enhanced transcription in SN48 cells, indicating that these opposite activities are intrinsic to Deaf-1. Repressor or enhancer activities of Deaf-1 or Gal4DBD-Deaf-1 were blocked by histone deacetylase inhibitor trichostatin A. Thus, the intrinsic activity of Deaf-1 at the 5-HT1A promoter is opposite in presynaptic versus postsynaptic neuronal cells and requires deacetylation. Cell specific regulation by Deaf-1 could underlie region-specific alterations in 5 HT1A receptor expression in different mood disorders. PMID- 16467536 TI - Increased vulnerability to nicotine self-administration and relapse in alcohol naive offspring of rats selectively bred for high alcohol intake. AB - The prevalence of smoking in human alcoholics is substantially higher than in the general population, and results from twin studies suggest that a shared genetic vulnerability underlies alcohol and nicotine addiction. Here, we directly tested this hypothesis by examining nicotine-taking behavior in alcohol-naive offspring of alcohol-preferring (P) rats and alcohol-nonpreferring (NP) rats that had been selectively bred for high and low alcohol intake. The self-administration of intravenous nicotine (0.015-0.060 mg/kg per infusion) in P rats was more than twice than that of NP rats. Nicotine seeking induced by reexposure to nicotine cues in extinction tests was also substantially greater in P rats than in NP rats. In a subsequent relapse test, priming nicotine injections reinstated drug seeking in P rats but not NP rats. P rats also self-administered higher amounts of oral sucrose (1-20%) than NP rats, a finding consistent with previous reports. In contrast, self-administration of intravenous cocaine (0.1875-1.125 mg/kg per infusion) was remarkably similar in the P and NP rats; however, P-NP differences in cocaine seeking emerged in subsequent extinction and cocaine priming-induced reinstatement tests. In both cases, lever responding was higher in P rats than in NP rats. Thus, alcohol-naive offspring of rats genetically selected for high alcohol intake are highly susceptible to nicotine self-administration and relapse, and this susceptibility is not likely caused by general reward deficits in NP rats. The present findings provide experimental evidence for the hypothesis that a shared genetic determinant accounts for the co-abuse of nicotine and alcohol. PMID- 16467537 TI - Astrocytes induce hemeoxygenase-1 expression in microglia: a feasible mechanism for preventing excessive brain inflammation. AB - Microglia are the major inflammatory cells in the brain, in which microglial inflammatory responses are modulated by interactions with other brain cells. Here, we show that astrocytes, the most abundant cells in the brain, can secrete one or more factors capable of modulating microglial activation by regulating the microglial levels of reactive oxygen species (ROS). Treatment of microglia with astrocyte culture-conditioned media (ACM) increased the expression level and activity of hemeoxygenase-1 (HO-1). ACM also induced nuclear translocation of the nuclear factor E2-related factor 2 transcription factor, increased the binding activity of the antioxidant response element (ARE), and enhanced HO-1 promoter activity in an ARE-dependent manner. Furthermore, treatment with ACM suppressed interferon-gamma (IFN-gamma)-induced ROS production, leading to reduced inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) release. In agreement with these results, mimickers of HO-1 products, such as bilirubin, ferrous iron, and a carbon monoxide-releasing molecule, reduced IFN-gamma-induced iNOS expression and/or NO release. Finally, we found that the active component(s) in ACM was heat labile and smaller than 3 kDa. Together, these results suggest that astrocytes could cooperate with microglia to prevent excessive inflammatory responses in the brain by regulating microglial expression of HO-1 and production of ROS. PMID- 16467538 TI - Images in clinical medicine. Subacute urinary retention. PMID- 16467539 TI - Facing the diabetes epidemic--mandatory reporting of glycosylated hemoglobin values in New York City. PMID- 16467540 TI - The demise of the physical exam. PMID- 16467541 TI - The stethoscope and the art of listening. PMID- 16467542 TI - Stroke and neurovascular protection. PMID- 16467543 TI - Saw palmetto for benign prostatic hyperplasia. AB - BACKGROUND: Saw palmetto is used by over 2 million men in the United States for the treatment of benign prostatic hyperplasia and is commonly recommended as an alternative to drugs approved by the Food and Drug Administration. METHODS: In this double-blind trial, we randomly assigned 225 men over the age of 49 years who had moderate-to-severe symptoms of benign prostatic hyperplasia to one year of treatment with saw palmetto extract (160 mg twice a day) or placebo. The primary outcome measures were changes in the scores on the American Urological Association Symptom Index (AUASI) and the maximal urinary flow rate. Secondary outcome measures included changes in prostate size, residual urinary volume after voiding, quality of life, laboratory values, and the rate of reported adverse effects. RESULTS: There was no significant difference between the saw palmetto and placebo groups in the change in AUASI scores (mean difference, 0.04 point; 95 percent confidence interval, -0.93 to 1.01), maximal urinary flow rate (mean difference, 0.43 ml per minute; 95 percent confidence interval, -0.52 to 1.38), prostate size, residual volume after voiding, quality of life, or serum prostate specific antigen levels during the one-year study. The incidence of side effects was similar in the two groups. CONCLUSIONS: In this study, saw palmetto did not improve symptoms or objective measures of benign prostatic hyperplasia. (ClinicalTrials.gov number, NCT00037154.). PMID- 16467544 TI - Radiotherapy plus cetuximab for squamous-cell carcinoma of the head and neck. AB - BACKGROUND: We conducted a multinational, randomized study to compare radiotherapy alone with radiotherapy plus cetuximab, a monoclonal antibody against the epidermal growth factor receptor, in the treatment of locoregionally advanced squamous-cell carcinoma of the head and neck. METHODS: Patients with locoregionally advanced head and neck cancer were randomly assigned to treatment with high-dose radiotherapy alone (213 patients) or high-dose radiotherapy plus weekly cetuximab (211 patients) at an initial dose of 400 mg per square meter of body-surface area, followed by 250 mg per square meter weekly for the duration of radiotherapy. The primary end point was the duration of control of locoregional disease; secondary end points were overall survival, progression-free survival, the response rate, and safety. RESULTS: The median duration of locoregional control was 24.4 months among patients treated with cetuximab plus radiotherapy and 14.9 months among those given radiotherapy alone (hazard ratio for locoregional progression or death, 0.68; P=0.005). With a median follow-up of 54.0 months, the median duration of overall survival was 49.0 months among patients treated with combined therapy and 29.3 months among those treated with radiotherapy alone (hazard ratio for death, 0.74; P=0.03). Radiotherapy plus cetuximab significantly prolonged progression-free survival (hazard ratio for disease progression or death, 0.70; P=0.006). With the exception of acneiform rash and infusion reactions, the incidence of grade 3 or greater toxic effects, including mucositis, did not differ significantly between the two groups. CONCLUSIONS: Treatment of locoregionally advanced head and neck cancer with concomitant high-dose radiotherapy plus cetuximab improves locoregional control and reduces mortality without increasing the common toxic effects associated with radiotherapy to the head and neck. (ClinicalTrials.gov number, NCT00004227.) PMID- 16467545 TI - Selective serotonin-reuptake inhibitors and risk of persistent pulmonary hypertension of the newborn. AB - BACKGROUND: Persistent pulmonary hypertension of the newborn (PPHN) is associated with substantial infant mortality and morbidity. A previous cohort study suggested a possible association between maternal use of the selective serotonin reuptake inhibitor (SSRI) fluoxetine late in the third trimester of pregnancy and the risk of PPHN in the infant. We performed a case-control study to assess whether PPHN is associated with exposure to SSRIs during late pregnancy. METHODS: Between 1998 and 2003, we enrolled 377 women whose infants had PPHN and 836 matched control women and their infants. Maternal interviews were conducted by nurses, who were blinded to the study hypothesis, regarding medication use in pregnancy and potential confounders, including demographic variables and health history. RESULTS: Fourteen infants with PPHN had been exposed to an SSRI after the completion of the 20th week of gestation, as compared with six control infants (adjusted odds ratio, 6.1; 95 percent confidence interval, 2.2 to 16.8). In contrast, neither the use of SSRIs before the 20th week of gestation nor the use of non-SSRI antidepressant drugs at any time during pregnancy was associated with an increased risk of PPHN. CONCLUSIONS: These data support an association between the maternal use of SSRIs in late pregnancy and PPHN in the offspring; further study of this association is warranted. These findings should be taken into account in decisions as to whether to continue the use of SSRIs during pregnancy. PMID- 16467547 TI - Clinical practice. Antibiotic allergy. PMID- 16467548 TI - The many roles of chemokines and chemokine receptors in inflammation. PMID- 16467549 TI - Images in clinical medicine. Radiation recall reaction after antimicrobial therapy. PMID- 16467550 TI - Case records of the Massachusetts General Hospital. Case 4-2006. A 79-year-old woman with myalgias, fatigue, and shortness of breath. PMID- 16467546 TI - NXY-059 for acute ischemic stroke. AB - BACKGROUND: NXY-059 is a free-radical-trapping agent that is neuroprotective in animal models of stroke. We tested whether it would reduce disability in humans after acute ischemic stroke. METHODS: We conducted a randomized, double-blind, placebo-controlled trial involving 1722 patients with acute ischemic stroke who were randomly assigned to receive a 72-hour infusion of placebo or intravenous NXY-059 within 6 hours after the onset of the stroke. The primary outcome was disability at 90 days, as measured according to scores on the modified Rankin scale for disability (range, 0 to 5, with 0 indicating no residual symptoms and 5 indicating bedbound, requiring constant care). RESULTS: Among the 1699 subjects included in the efficacy analysis, NXY-059 significantly improved the overall distribution of scores on the modified Rankin scale, as compared with placebo (P=0.038 by the Cochran-Mantel-Haenszel test). The common odds ratio for improvement across all categories of the scale was 1.20 (95 percent confidence interval, 1.01 to 1.42). Mortality and rates of serious and nonserious adverse events were each similar in the two groups. NXY-059 did not improve neurologic functioning as measured according to the National Institutes of Health Stroke Scale (NIHSS): the difference between the two groups in the change from baseline scores was 0.1 point (95 percent confidence interval, -1.4 to 1.1; P=0.86). Likewise, no improvement was observed according to the Barthel index (P=0.14). In a post hoc analysis of patients who also received alteplase, NXY-059 was associated with a lower incidence of any hemorrhagic transformation (P=0.001) and symptomatic intracranial hemorrhage (P=0.036). CONCLUSIONS: The administration of NXY-059 within six hours after the onset of acute ischemic stroke significantly improved the primary outcome (reduced disability at 90 days), but it did not significantly improve other outcome measures, including neurologic functioning as measured by the NIHSS score. Additional research is needed to confirm whether NXY 059 is beneficial in ischemic stroke. (ClinicalTrials.gov number, NCT00119626.). PMID- 16467551 TI - Proven and unproven therapy for benign prostatic hyperplasia. PMID- 16467552 TI - Cetuximab and radiotherapy for head and neck cancer. PMID- 16467553 TI - Depressing observations on the use of selective serotonin-reuptake inhibitors during pregnancy. PMID- 16467555 TI - Trastuzumab in breast cancer. PMID- 16467557 TI - Long-acting methods of contraception. PMID- 16467556 TI - Selective adhesion-molecule therapy and inflammatory bowel disease. PMID- 16467558 TI - Medical management of depression. PMID- 16467559 TI - Ventricular tachycardia associated with trastuzumab. PMID- 16467560 TI - The portability of tagSNPs across populations: a worldwide survey. AB - In the search for common genetic variants that contribute to prevalent human diseases, patterns of linkage disequilibrium (LD) among linked markers should be considered when selecting SNPs. Genotyping efficiency can be increased by choosing tagging SNPs (tagSNPs) in LD with other SNPs. However, it remains to be seen whether tagSNPs defined in one population efficiently capture LD in other populations; that is, how portable tagSNPs are. Indeed, tagSNP portability is a challenge for the applicability of HapMap results. We analyzed 144 SNPs in a 1-Mb region of chromosome 22 in 1055 individuals from 38 worldwide populations, classified into seven continental groups. We measured tagSNP portability by choosing three reference populations (to approximate the three HapMap populations), defining tagSNPs, and applying them to other populations independently on the availability of information on the tagSNPs in the compared population. We found that tagSNPs are highly informative in other populations within each continental group. Moreover, tagSNPs defined in Europeans are often efficient for Middle Eastern and Central/South Asian populations. TagSNPs defined in the three reference populations are also efficient for more distant and differentiated populations (Oceania, Americas), in which the impact of their special demographic history on the genetic structure does not interfere with successfully detecting the most common haplotype variation. This high degree of portability lends promise to the search for disease association in different populations, once tagSNPs are defined in a few reference populations like those analyzed in the HapMap initiative. PMID- 16467561 TI - Analysis of allelic differential expression in human white blood cells. AB - Allelic variation of gene expression is common in humans, and is of interest because of its potential contribution to variation in heritable traits. To identify human genes with allelic expression differences, we genotype DNA and examine mRNA isolated from the white blood cells of 12 unrelated individuals using oligonucleotide arrays containing 8406 exonic SNPs. Of the exonic SNPs, 1983, located in 1389 genes, are both expressed in the white blood cells and heterozygous in at least one of the 12 individuals, and thus can be examined for differential allelic expression. Of the 1389 genes, 731 (53%) show allele expression differences in at least one individual. To gain insight into the regulatory mechanisms governing allelic expression differences, we analyze a set of 60 genes containing exonic SNPs that are heterozygous in three or more samples, and for which all heterozygotes display differential expression. We find three patterns of allelic expression, suggesting different underlying regulatory mechanisms. Exonic SNPs in three of the 60 genes are monoallelically expressed in the human white blood cells, and when examined in families show expression of only the maternal copy, consistent with regulation by imprinting. Approximately one-third of the genes have the same allele expressed more highly in all heterozygotes, suggesting that their regulation is predominantly influenced by cis-elements in strong linkage disequilibrium with the assayed exonic SNP. The remaining two-thirds of the genes have different alleles expressed more highly in different heterozygotes, suggesting that their expression differences are influenced by factors not in strong linkage disequilibrium with the assayed exonic SNP. PMID- 16467562 TI - Training-induced functional activation changes in dual-task processing: an FMRI study. AB - Although training-induced changes in brain activity have been previously examined, plasticity associated with executive functions remains understudied. In this study, we examined training-related changes in cortical activity during a dual task requiring executive control. Two functional magnetic resonance imaging (fMRI) sessions, one before training and one after training, were performed on both a control group and a training group. Using a region-of-interest analysis, we examined Time x Group and Time x Group x Condition interactions to isolate training-dependent changes in activation. We found that most regions involved in dual-task processing before training showed reductions in activation after training. Many of the decreases in activation were correlated with improved performance on the task. We also found an area in the dorsolateral prefrontal cortex that showed an increase in activation for the training group for the dual task condition, which was also correlated with improved performance. These results are discussed in relation to the efficacy of training protocols for modulating attention and executive functions, dual-task processing, and fMRI correlates of plasticity. PMID- 16467563 TI - Immunocytochemical distribution of the cannabinoid CB1 receptor in the primate neocortex: a regional and laminar analysis. AB - Delta-9-tetrahydrocannabinol (Delta9-THC) has profound effects on higher cognitive functions, and exposure to Delta9-THC has been associated with the appearance or exacerbation of the clinical features of schizophrenia. These actions appear to be mediated via the CB1 receptor, the principal cannabinoid receptor expressed in the brain. However, the distribution of the CB1 receptor in neocortical regions of the primate brain that mediate cognitive functions is not known. We therefore investigated the immunocytochemical localization of the CB1 receptor in the brains of macaque monkeys and humans using antibodies that specifically recognize the N- or C-terminus of the CB1 receptor. In monkeys, intense CB1 immunoreactivity was observed primarily in axons and boutons. Across neocortical regions of the monkey brain, CB1-immunoreactive (IR) axons exhibited considerable heterogeneity in density and laminar distribution. Neocortical association regions, such as the prefrontal and cingulate cortices, demonstrated a higher density, and exhibited a unique laminar pattern of CB1-IR axons, compared with primary sensory and motor cortices. Similar regional and laminar distributions of CB1-IR axons were also present in the human neocortex. CB1-IR axons had more prominent varicosities in human tissue, but this difference appeared to represent a postmortem effect as similar morphological features increased in unperfused monkey tissue as a function of postmortem interval. In electron microscopy studies of perfused monkey prefrontal cortex, CB1 immunoreactivity was predominantly found in axon terminals that exclusively formed symmetric synapses. The high density, distinctive laminar distribution, and localization to inhibitory terminals of CB1 receptors in primate higher-order association regions suggests that the CB1 receptor may play a critical role in the circuitry that subserves cognitive functions such as those that are disturbed in schizophrenia. PMID- 16467564 TI - Differential effects of endocannabinoids on glutamatergic and GABAergic inputs to layer 5 pyramidal neurons. AB - Endocannabinoids are emerging as potent modulators of neuronal activity throughout the brain, and activation of the type-1 cannabinoid receptor (CB1R) reduces sensory-evoked cortical responses in vivo, presumably by decreasing excitatory transmission. In the neocortex, CB1R is differentially expressed across neocortical laminae, with highest levels of expression in layers 2/3 and 5. Although we have shown that cannabinoid signaling in layer 2/3 of somatosensory cortex targets both gamma-aminobutyric acid (GABA) and glutamate release, the predominant effect is a net increase in pyramidal neuron (PN) activity due to disinhibition. The role of endocannabinoid signaling in layer 5, the main output layer of the neocortex, remains unknown. We found that inducing activity in layer 5 PNs resulted in endocannabinoid-mediated depolarization induced suppression of excitation (DSE), whereas the majority of inhibitory inputs were cannabinoid insensitive. Furthermore, in contrast to layer 2/3, the net effect of elevations in action potential firing of layer 5 PNs was an endocannabinoid-mediated decrease in PN spike probability. Interestingly, excitatory synaptic currents in layer 5 evoked by intralaminar stimulation were cannabinoid sensitive, whereas inputs evoked from layer 2/3 were insensitive, suggesting specificity of cannabinoid signaling across glutamatergic inputs. Thus, cannabinoids have differential effects on excitation and inhibition across cortical layers, and endocannabinoid signaling in layer 5 may serve to selectively decrease the efficacy of a subset of excitatory inputs. PMID- 16467565 TI - V2 thin stripes contain spatially organized representations of achromatic luminance change. AB - A considerable amount of research over the last decades has focused on the apparent specialization of V2 thin stripes for the processing of color in diurnal primates. However, because V2 thin stripes are functionally heterogeneous in that they consist of largely separate color- and luminance-preferring domains and because the color-preferring domains contain a systematic representation of hue, we hypothesized that they contained functional maps that subserve luminance processing. Here we show, using optical imaging of intrinsic cortical signals and microelectrode recording, that the V2 thin stripe luminance-preferring domains contain spatially segregated modules that encode the direction of relative luminance change. Quantitative analysis of the cortical responses to luminance increments or decrements indicates that these luminance-sensitive modules also encode the magnitude of the luminance change by the magnitude of the evoked cortical response. These results demonstrate an important role of V2 thin stripes in the processing of luminance and thus suggest that thin stripes are involved in the overall processing of the surface properties of objects rather than simply the processing of color. PMID- 16467566 TI - Electrophysiological markers of visuocortical development. AB - The development of noninvasive techniques for the assessment of functional brain maturation is critical. The present study analyzed 63 babies' and children's (27 days to 5.5 years) cerebral responses to a pattern-reversal visual stimulation using high-density (128 electrodes) electrophysiological recordings. Developmental data were further compared with those of young adults (n = 16). Tremendous changes in pattern visual evoked potentials (pVEPs) morphology were observed between 7 and 24 months characterized by the emergence of negative components labeled "N70" and "N145" and the reduction of the P100 amplitude. The adult pattern of response appears from 24 months onward. Spectral density values show an increase of higher frequencies with age. Coherence values show a reduction between 3 and 23 months of age as well as a further increase toward adulthood between areas implicated in visual processing. These results are discussed in light of developmental features such as synaptogenesis, myelination, and neuronal networks refinement. PMID- 16467567 TI - Correlation between axonal morphologies and synaptic input kinetics of interneurons from mouse visual cortex. AB - Neocortical interneurons display great morphological and physiological variability and are ideally positioned to control circuit dynamics, although their exact role is still poorly understood. To better understand this diversity, we have performed a detailed anatomical and physiological characterization of 3 subtypes of visual cortex interneurons, isolated from transgenic mice which express green fluorescent protein in somatostatin, parvalbumin, and neuropeptide Y positive neurons. We find that these 3 groups of interneurons have systematic differences in dendritic and axonal morphologies and also characteristically differ in the frequencies, amplitude, and kinetics of the spontaneous excitatory and inhibitory synaptic currents they receive. Moreover, we detect a correlation between the kinetics of their synaptic inputs and quantitative aspects of their axonal arborizations. This suggests that different interneuron types could channel different temporal patterns of activity. Our results also confirm the importance of the axonal morphology to classify interneurons. PMID- 16467568 TI - Multiparametric changes in the receptive field of cortical auditory neurons induced by thalamic activation in the mouse. AB - The functional organization of the sensory cortex is constructed to process sensory information based on experience and learning. Importantly, it is plastic so that it can quickly adapt to environmental changes. Because the thalamus gates all ascending information, it is critical to understand how the thalamocortical system contributes to the plasticity of the sensory cortex. We show here that the neuronal receptive field (RF) in the auditory cortex faithfully tends toward the RF of the electrically stimulated auditory thalamic neurons. We characterized the RF of auditory neurons by measuring the best frequency, minimum threshold, bandwidth, RF area, and averaged response magnitude. All these parameters of the cortical RF showed robust changes toward the values of the parameters of the stimulated thalamic neuron following focal thalamic stimulation. Our data suggest that the thalamocortical system possesses intrinsic mechanisms that underlie the input specificity of learning-induced or experience-dependent cortical plasticity. PMID- 16467569 TI - Regulation of membrane traffic by phosphoinositide 3-kinases. AB - Phosphoinositide (PI) 3-kinases control essential cellular functions such as cytoskeletal dynamics, signal transduction and membrane trafficking. FYVE, PX and PH domains mediate the binding of effector proteins to the lipid products of PI 3 kinases. Recent studies have provided significant insights into the roles of PI 3 kinases, their catalytic products and their downstream effectors in membrane trafficking. Class I and II PI 3-kinases trigger receptor-induced trafficking processes, such as phagocytosis, macropinocytosis and regulated exocytosis. Class I PI 3-kinases also function to inhibit autophagy. By contrast, class III PI 3 kinases mainly mediate receptor-independent trafficking events, which mostly are related to endocytic membrane traffic, phagosome maturation and autophagy. PMID- 16467570 TI - Beyond lectins: the calnexin/calreticulin chaperone system of the endoplasmic reticulum. AB - Calnexin and calreticulin are related proteins that comprise an ER chaperone system that ensures the proper folding and quality control of newly synthesized glycoproteins. The specificity for glycoproteins is conferred by a lectin site that recognizes an early oligosaccharide processing intermediate on the folding glycoprotein, Glc1Man9GlcNAc2. In addition, calnexin and calreticulin possess binding sites for ATP, Ca2+, non-native polypeptides and ERp57, an enzyme that catalyzes disulfide bond formation, reduction and isomerization. Recent studies have revealed the locations of some of these ligand-binding sites and have provided insights into how they contribute to overall chaperone function. In particular, the once controversial non-native-polypeptide-binding site has now been shown to function both in vitro and in cells. Furthermore, there is clear evidence that ERp57 participates in glycoprotein biogenesis either alone or in tandem with calnexin and calreticulin. PMID- 16467571 TI - Glycosylation catalyzed by lysyl hydroxylase 3 is essential for basement membranes. AB - Lysyl hydroxylase 3 (LH3) is a multifunctional enzyme possessing lysyl hydroxylase (LH), hydroxylysyl galactosyltransferase (GT) and galactosylhydroxylysyl glucosyltransferase (GGT) activities in vitro. To investigate the in vivo importance of LH3-catalyzed lysine hydroxylation and hydroxylysine-linked glycosylations, three different LH3-manipulated mouse lines were generated. Mice with a mutation that blocked only the LH activity of LH3 developed normally, but showed defects in the structure of the basement membrane and in collagen fibril organization in newborn skin and lung. Analysis of a hypomorphic LH3 mouse line with the same mutation, however, demonstrated that the reduction of the GGT activity of LH3 disrupts the localization of type IV collagen, and thus the formation of basement membranes during mouse embryogenesis leading to lethality at embryonic day (E) 9.5-14.5. Strikingly, survival of hypomorphic embryos and the formation of the basement membrane were directly correlated with the level of GGT activity. In addition, an LH3-knockout mouse lacked GGT activity leading to lethality at E9.5. The results confirm that LH3 has LH and GGT activities in vivo, LH3 is the main molecule responsible for GGT activity and that the GGT activity, not the LH activity of LH3, is essential for the formation of the basement membrane. Together our results demonstrate for the first time the importance of hydroxylysine-linked glycosylation for collagens. PMID- 16467572 TI - Activity of the hSPCA1 Golgi Ca2+ pump is essential for Ca2+-mediated Ca2+ response and cell viability in Darier disease. AB - Keratinocyte differentiation, adhesion and motility are directed by extracellular Ca2+ concentration increases, which in turn increase intracellular Ca2+ levels. Normal keratinocytes, in contrast to most non-excitable cells, require Ca2+ release from both Golgi and endoplasmic reticulum Ca2+ stores for efficient Ca2+ signaling. Dysfunction of the Golgi human secretory pathway Ca2+-ATPase hSPCA1, encoded by ATP2C1, abrogates Ca2+ signaling and causes the acantholytic genodermatosis, Hailey-Hailey disease. We have examined the role of the endoplasmic reticulum Ca2+ store, established and maintained by the sarcoplasmic and endoplasmic reticulum Ca2+-ATPase SERCA2 encoded by ATP2A2, in Ca2+ signaling. Although previous studies have shown acute SERCA2 inactivation to abrogate Ca2+ signaling, we find that chronic inactivation of ATP2A2 in keratinocytes from patients with the similar acantholytic genodermatosis, Darier disease, does not impair the response to raised extracellular Ca2+ levels. This normal response is due to a compensatory upregulation of hSPCA1, as inactivating ATP2C1 expression with siRNA blocks the response to raised extracellular Ca2+ concentrations in both normal and Darier keratinocytes. ATP2C1 inactivation also diminishes Darier disease keratinocyte viability, suggesting that compensatory ATP2C1 upregulation maintains viability and partially compensates for defective endoplasmic reticulum Ca2+-ATPase in Darier disease keratinocytes. Keratinocytes thus are unique among mammalian cells in their ability to use the Golgi Ca2+ store to mediate Ca2+ signaling. PMID- 16467574 TI - The use of a simulated body shape for determination of patient dosimetry within whole body ultraviolet treatment cabinets. AB - Details are described of a simulated body shape utilized to determine patient dosimetry within whole body ultraviolet treatment cabinets. The body shape was designed to have a shape characteristic of the patient group undergoing treatment and was utilized in several Waldmann whole body treatment cabinets and with irradiance measurements undertaken using a Bentham DM150C spectroradiometer. It was considered that the rigid defined structure of the body shape allowed satisfactory reproducibility of measurements in such cabinets and also allowed additional parameters to be identified for evaluation of performance of such units. PMID- 16467575 TI - Determination of eye safety filter protection factors associated with retinal thermal hazard and blue light photochemical hazard for intense pulsed light sources. AB - An assessment is provided of protection factors afforded for retinal thermal hazard and blue light photochemical hazard for a range of filters used with intense pulsed light sources (IPLs). A characteristic IPL spectrum based on black body radiation at 5000 K with a low cut filter at 515 nm was identified as suitable for such estimations. Specific filters assessed included types with idealized transmission properties and also a range of types whose transmission characteristics were measured by means of a Bentham DMc150 spectroradiometer. Predicted behaviour based on these spectra is outlined which describes both the effectiveness of protection and the level of luminous transmittance afforded. The analysis showed it was possible to describe a figure of merit for a particular filter material relating the degree of protection provided and corresponding value of luminous transmittance. This consideration is important for providing users of IPL equipment with safety eyewear with adequate level of visual transmittance. PMID- 16467576 TI - Verification of a rounded leaf-end MLC model used in a radiotherapy treatment planning system. AB - A new multileaf collimator (MLC) model has been incorporated into version 7.4 of the Pinnacle radiotherapy treatment planning system (Philips Radiation Oncology Systems, Milpitas, CA). The MLC model allows for rounded MLC leaf-ends and provides separate parameters for inter-leaf transmission, intra-leaf transmission and the tongue width of the MLC leaf. In this report we detail the method followed to commission the MLC model for a Varian 120-leaf Millennium MLC (Varian Medical Systems, Palo Alto, CA, USA) for both 6 and 10 MV photons, and test the validity of the model for an IMRT field. Dose profiles in water were measured for a range of square MLC field sizes and compared to the Pinnacle computed dose profiles; in addition, the dose distribution for a series of adjacent MLC fields was measured to observe the model's behaviour along match-lines. Based on these results intra-leaf transmissions of 1.5% for 6 MV and 1.8% for 10 MV, leaf-tip radius of 12.0 cm, an inter-leaf transmission of 0.5%, and a tongue width of 0.1 cm were chosen. Using these values to compute the planar dose distribution for a 6 MV IMRT field, the new version of Pinnacle displayed improved dosimetric agreement with the dose-to-water EPID image and ion chamber measurements when compared to the old version of Pinnacle, particularly along the MLC tongue edge and across match-lines. Discrepancies of up to 5% were observed between calculated and measured doses along match-lines for both 6 MV and 10 MV photons; however, the new MLC model did predict the presence of match-lines and was a significant improvement on the previous model. PMID- 16467577 TI - On dose distribution comparison. AB - In radiotherapy practice, one often needs to compare two dose distributions. Especially with the wide clinical implementation of intensity-modulated radiation therapy, software tools for quantitative dose (or fluence) distribution comparison are required for patient-specific quality assurance. Dose distribution comparison is not a trivial task since it has to be performed in both dose and spatial domains in order to be clinically relevant. Each of the existing comparison methods has its own strengths and weaknesses and there is room for improvement. In this work, we developed a general framework for comparing dose distributions. Using a new concept called maximum allowed dose difference (MADD), the comparison in both dose and spatial domains can be performed entirely in the dose domain. Formulae for calculating MADD values for various comparison methods, such as composite analysis and gamma index, have been derived. For convenience in clinical practice, a new measure called normalized dose difference (NDD) has also been proposed, which is the dose difference at a point scaled by the ratio of MADD to the predetermined dose acceptance tolerance. Unlike the simple dose difference test, NDD works in both low and high dose gradient regions because it considers both dose and spatial acceptance tolerances through MADD. The new method has been applied to a test case and a clinical example. It was found that the new method combines the merits of the existing methods (accurate, simple, clinically intuitive and insensitive to dose grid size) and can easily be implemented into any dose/intensity comparison tool. PMID- 16467578 TI - Dynamic ventilation imaging from four-dimensional computed tomography. AB - A novel method for dynamic ventilation imaging of the full respiratory cycle from four-dimensional computed tomography (4D CT) acquired without added contrast is presented. Three cases with 4D CT images obtained with respiratory gated acquisition for radiotherapy treatment planning were selected. Each of the 4D CT data sets was acquired during resting tidal breathing. A deformable image registration algorithm mapped each (voxel) corresponding tissue element across the 4D CT data set. From local average CT values, the change in fraction of air per voxel (i.e. local ventilation) was calculated. A 4D ventilation image set was calculated using pairs formed with the maximum expiration image volume, first the exhalation then the inhalation phases representing a complete breath cycle. A preliminary validation using manually determined lung volumes was performed. The calculated total ventilation was compared to the change in contoured lung volumes between the CT pairs (measured volume). A linear regression resulted in a slope of 1.01 and a correlation coefficient of 0.984 for the ventilation images. The spatial distribution of ventilation was found to be case specific and a 30% difference in mass-specific ventilation between the lower and upper lung halves was found. These images may be useful in radiotherapy planning. PMID- 16467579 TI - Targeted disruption of the blood-brain barrier with focused ultrasound: association with cavitation activity. AB - Acoustic emission was monitored during focused ultrasound exposures in conjunction with an ultrasound contrast agent (Optison) in order to determine if cavitation activity is associated with the induction of blood-brain barrier disruption (BBBD). Thirty-four locations were sonicated (frequency: 260 kHz) at targets 10 mm deep in rabbit brain (N = 9). The sonications were applied at peak pressure amplitudes ranging from 0.11 to 0.57 MPa (burst length: 10 ms; repetition frequency of 1 Hz; duration: 20 s). Acoustic emission was recorded with a focused passive cavitation detector. This emission was recorded at each location during sonications with and without Optison. Detectable wideband acoustic emission was observed only at 0.40 and 0.57 MPa. BBBD was observed in contrast MRI after sonication at 0.29-0.57 MPa. The appearance of small regions of extravasated erythrocytes appeared to be associated with this wideband emission signal. The results thus suggest that BBBD resulting from focused ultrasound pulses in the presence of Optison can occur without indicators for inertial cavitation in vivo, wideband emission and extravasation. If inertial cavitation is not responsible for the BBBD, other ultrasound/microbubble interactions are likely the source. A significant increase in the emission signal due to Optison at the second and third harmonics of the ultrasound driving frequency was found to correlate with BBBD and might be useful as an online method to indicate when the disruption occurs. PMID- 16467580 TI - Towards patient specific thermal modelling of the prostate. AB - The application of thermal modelling for hyperthermia and thermal ablation is severely hampered by lack of information about perfusion and vasculature. However, recently, with the advent of sophisticated angiography and dynamic contrast enhanced (DCE) imaging techniques, it has become possible to image small vessels and blood perfusion bringing the ultimate goal of patient specific thermal modelling closer within reach. In this study dynamic contrast enhanced multi-slice CT imaging techniques are employed to investigate the feasibility of this concept for regional hyperthermia treatment of the prostate. The results are retrospectively compared with clinical thermometry data of a patient group from an earlier trial. Furthermore, the role of the prostate vasculature in the establishment of the prostate temperature distribution is studied. Quantitative 3D perfusion maps of the prostate were constructed for five patients using a distributed-parameter tracer kinetics model to analyse dynamic CT data. CT angiography was applied to construct a discrete vessel model of the pelvis. Additionally, a discrete vessel model of the prostate vasculature was constructed of a prostate taken from a human corpse. Three thermal modelling schemes with increasing inclusion of the patient specific physiological information were used to simulate the temperature distribution of the prostate during regional hyperthermia. Prostate perfusion was found to be heterogeneous and T3 prostate carcinomas are often characterized by a strongly elevated tumour perfusion (up to 70-80 ml 100 g(-1) min(-1)). This elevated tumour perfusion leads to 1-2 degrees C lower tumour temperatures than thermal simulations based on a homogeneous prostate perfusion. Furthermore, the comparison has shown that the simulations with the measured perfusion maps result in consistently lower prostate temperatures than clinically achieved. The simulations with the discrete vessel model indicate that significant pre-heating takes place in the prostate capsule vasculature which forms a possible explanation for the discrepancy. Pre-heating in the larger pelvic vessels is very moderate, approximately 0.1-0.3 degrees C. In conclusion, perfusion imaging provides important input for thermal modelling and can be used to obtain a lower limit on the prostate and tumour temperature in regional hyperthermia. However, it is not sufficient to calculate in detail the prostate temperature distribution in individual patients. The prostate vasculature plays such a crucial role that a patient specific discrete vessel model of the prostate vasculature is required. PMID- 16467581 TI - Analysis of factors important for transurethral ultrasound prostate heating using MR temperature feedback. AB - The feasibility of using MR thermometry for temperature feedback to control a transurethral ultrasound heating applicator with planar transducers was investigated. The sensitivity of a temperature-based feedback algorithm to spatial (control point area, slice thickness, angular alignment) and non-spatial (imaging time, temperature uncertainty) parameters was evaluated through numerical simulations. The angular alignment of the control point with the ultrasound beam was an important parameter affecting the average spatial error in heat delivery. The other spatial parameters were less influential, thus providing an opportunity to reduce spatial resolution for increased SNR in the MR imaging. The update time was the most important non-spatial parameter determining the performance of the control algorithm. Combined non-spatial and spatial parameters achieved acceptable performance with a voxel size of 3 mm x 3 mm, a 10 mm slice thickness and a 5 s update time. Temperature uncertainty of up to 2 degrees C had little effect on the performance of the control algorithm but did reduce the average error slightly due to a systematic, noise-induced overestimation of the boundary temperature. These simulations imply that MR thermometry performed on clinical 1.5 T imaging systems is of sufficient quality for use as thermal feedback for conformal prostate thermal therapy with transurethral ultrasound heating applicators incorporating planar transducers. PMID- 16467582 TI - Evaluating microcirculation by pulsatile laser Doppler signal. AB - Laser Doppler flowmetry (LDF) is a popular method for monitoring the microcirculation, but it does not provide absolute measurements. Instead, the mean flux response or energy distribution in the frequency domain is generally compared before and after stimulus. Using the heartbeat as a trigger, we investigated whether the relation between pressure and flux can be used to discriminate different microcirculatory conditions. We propose the following three pulsatile indices for evaluating the microcirculation condition from the normalized pressure and flux segment with a synchronized-averaging method: peak delay time (PDT), pressure rise time and flux rise time (FRT). The abdominal aortic blood pressure and renal cortex flux (RCF) signals were measured in spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY). The mean value of the RCF did not differ between SHR and WKY. However, the PDT was longer in SHR (87.14 +/- 5.54 ms, mean +/- SD) than in WKY (76.92 +/- 2.62 ms; p < 0.001). The FRT was also longer in SHR (66.56 +/- 1.98 ms) than in WKY (58.02 +/- 1.77 ms; p < 0.001). We propose that a new dimension for comparing the LDF signals, which the results from the present study show, can be used to discriminate RCF signals that cannot be discriminated using traditional methods. PMID- 16467583 TI - A three-dimensional-weighted cone beam filtered backprojection (CB-FBP) algorithm for image reconstruction in volumetric CT-helical scanning. AB - Based on the structure of the original helical FDK algorithm, a three-dimensional (3D)-weighted cone beam filtered backprojection (CB-FBP) algorithm is proposed for image reconstruction in volumetric CT under helical source trajectory. In addition to its dependence on view and fan angles, the 3D weighting utilizes the cone angle dependency of a ray to improve reconstruction accuracy. The 3D weighting is ray-dependent and the underlying mechanism is to give a favourable weight to the ray with the smaller cone angle out of a pair of conjugate rays but an unfavourable weight to the ray with the larger cone angle out of the conjugate ray pair. The proposed 3D-weighted helical CB-FBP reconstruction algorithm is implemented in the cone-parallel geometry that can improve noise uniformity and image generation speed significantly. Under the cone-parallel geometry, the filtering is naturally carried out along the tangential direction of the helical source trajectory. By exploring the 3D weighting's dependence on cone angle, the proposed helical 3D-weighted CB-FBP reconstruction algorithm can provide significantly improved reconstruction accuracy at moderate cone angle and high helical pitches. The 3D-weighted CB-FBP algorithm is experimentally evaluated by computer-simulated phantoms and phantoms scanned by a diagnostic volumetric CT system with a detector dimension of 64 x 0.625 mm over various helical pitches. The computer simulation study shows that the 3D weighting enables the proposed algorithm to reach reconstruction accuracy comparable to that of exact CB reconstruction algorithms, such as the Katsevich algorithm, under a moderate cone angle (4 degrees) and various helical pitches. Meanwhile, the experimental evaluation using the phantoms scanned by a volumetric CT system shows that the spatial resolution along the z-direction and noise characteristics of the proposed 3D-weighted helical CB-FBP reconstruction algorithm are maintained very well in comparison to the FDK-type algorithms. Moreover, the experimental evaluation by clinical data verifies that the proposed 3D-weighted CB-FBP algorithm for image reconstruction in volumetric CT under helical source trajectory meets the challenges posed by diagnostic applications of volumetric CT imaging. PMID- 16467584 TI - Evaluation of accelerated iterative x-ray CT image reconstruction using floating point graphics hardware. AB - Statistical reconstruction methods offer possibilities to improve image quality as compared with analytical methods, but current reconstruction times prohibit routine application in clinical and micro-CT. In particular, for cone-beam x-ray CT, the use of graphics hardware has been proposed to accelerate the forward and back-projection operations, in order to reduce reconstruction times. In the past, wide application of this texture hardware mapping approach was hampered owing to limited intrinsic accuracy. Recently, however, floating point precision has become available in the latest generation commodity graphics cards. In this paper, we utilize this feature to construct a graphics hardware accelerated version of the ordered subset convex reconstruction algorithm. The aims of this paper are (i) to study the impact of using graphics hardware acceleration for statistical reconstruction on the reconstructed image accuracy and (ii) to measure the speed increase one can obtain by using graphics hardware acceleration. We compare the unaccelerated algorithm with the graphics hardware accelerated version, and for the latter we consider two different interpolation techniques. A simulation study of a micro-CT scanner with a mathematical phantom shows that at almost preserved reconstructed image accuracy, speed-ups of a factor 40 to 222 can be achieved, compared with the unaccelerated algorithm, and depending on the phantom and detector sizes. Reconstruction from physical phantom data reconfirms the usability of the accelerated algorithm for practical cases. PMID- 16467585 TI - Development of a tapping device: a new needle insertion method for prostate brachytherapy. AB - The purpose of this study is to develop and test a tapping device for needle insertion for prostate brachytherapy. This device will tap the needle into the prostate with a certain, well-defined, amount of momentum, instead of the currently used method of pushing the needle. Because of the high needle insertion velocity, we expect prostate motion and deformation to be less compared to current methods. We measured the momentum that is applied when manually tapping the needle into the prostate and found a mean momentum of 0.50 +/- 0.07 N s. The tapping device is pneumatically driven and we found that the delivered momentum increased linearly with the applied air pressure. The efficacy of the tapping device was tested on a piece of beef, placed on a freely moving and rotating platform. A significant correlation was found between the applied pressure and the rotation and displacement of the beef. Displacements and rotations were minimal for the highest pressure (4 bar) and amounted to only 2 mm and 6 degrees, respectively. Higher air pressures will further reduce displacements and rotations. PMID- 16467587 TI - Dose response evaluation of a low-density normoxic polymer gel dosimeter using MRI. AB - A low-density (approximately 0.6 g cm(-3)) normoxic polymer gel, containing the antioxidant tetrakis (hydroxymethyl) phosponium (THP), has been investigated with respect to basic absorbed dose response characteristics. The low density was obtained by mixing the gel with expanded polystyrene spheres. The depth dose data for 6 and 18 MV photons were compared with Monte Carlo calculations. A large volume phantom was irradiated in order to study the 3D dose distribution from a 6 MV field. Evaluation of the gel was carried out using magnetic resonance imaging. An approximately linear response was obtained for 1/T2 versus dose in the dose range of 2 to 8 Gy. A small decrease in the dose response was observed for increasing concentrations of THP. A good agreement between measured and Monte Carlo calculated data was obtained, both for test tubes and the larger 3D phantom. It was shown that a normoxic polymer gel with a reduced density could be obtained by adding expanded polystyrene spheres. In order to get reliable results, it is very important to have a uniform distribution of the gel and expanded polystyrene spheres in the phantom volume. PMID- 16467586 TI - Ion recombination correction in the Clatterbridge Centre of Oncology clinical proton beam. AB - Most codes of practice for dosimetry of proton beams do not give a clear recommendation on the determination of recombination correction factors for ionization chambers. In this work, recombination corrections were measured in the low-energy clinical proton beam of the Clatterbridge Centre of Oncology (CCO) using data collected at different dose rates and different polarizing voltages. This approach allows the separation of contributions from initial and volume recombination and was compared with results from extrapolation and two-voltage methods. A modified formulation of the method is presented for a modulated beam in which the ionization current is time dependent. Using a set-up with two identical chambers placed face-to-face yielded highly accurate data for plane parallel ionization chambers. This method may also be used for high-energy photon and electron beam dosimetry. At typical dose rates of 26 Gy min(-1) used clinically at the CCO, the recombination correction is 0.8% and thus is of importance for reference dosimetry. The proton beam should be treated as purely continuous given the high pulse repetition frequency of the cyclotron beam. The results show that the volume recombination parameter for protons is consistent with values measured for photon beams. Initial recombination was found to be independent of beam quality, except for a tendency to increase at the distal edge of the Bragg peak; this is only relevant for depth dose measurements. Using a general equation for recombination and generic values for the initial and volume recombination parameters (A = 0.25 V and m2 = 3.97 x 10(3) s cm(-1) nC(-1) V2), the experimental results are reproduced within 0.1% for all conditions met in this work. For the CCO beam and similar proton beams used for treating optical targets operating at high dose rates, the recombination correction factor can be overestimated by up to 2%, resulting in an overestimation of dose to water by the same amount, if the recommendation from IAEA TRS-398, which is only valid for pulsed beams, is followed without consideration. PMID- 16467588 TI - A new approach to the determination of air kerma using primary-standard cavity ionization chambers. AB - A consistent formalism is presented using Monte Carlo calculations to determine the reference air kerma from the measured energy deposition in a primary-standard cavity ionization chamber. A global approach avoiding the use of cavity ionization theory is discussed and its limitations shown in relation to the use of the recommended value for W. The role of charged-particle equilibrium is outlined and the consequent requirements placed on the calculations are detailed. Values for correction factors are presented for the BIPM air-kerma standard for 60Co, making use of the Monte Carlo code PENELOPE, a detailed geometrical model of the BIPM 60Co source and event-by-event electron transport. While the wall correction factor k(wall) = 1.0012(2) is somewhat lower than the existing value, the axial non-uniformity correction k(an) = 1.0027(3) is significantly higher. The use of a point source in the evaluation of k(an) is discussed. A comparison is made of the calculated dose ratio with the Bragg-Gray and Spencer-Attix stopping-power ratios, the results indicating a preference for the Bragg-Gray approach in this particular case. A change to the recommended value for W of up to 2 parts in 10(3) is discussed. The uncertainties arising from the geometrical models, the use of phase-space files, the radiation transport algorithms and the underlying radiation interaction coefficients are estimated. PMID- 16467590 TI - Positron autoradiography for intravascular imaging: feasibility evaluation. AB - Approximately 70% of acute coronary artery disease is caused by unstable (vulnerable) plaques with an inflammation of the overlying cap and high lipid content. A rupturing of the inflamed cap of the plaque results in propagation of the thrombus into the lumen, blockage of the artery and acute ischaemic syndrome or sudden death. Morphological imaging such as angiography or intravascular ultrasound cannot determine inflammation status of the plaque. A radiotracer such as 18F-FDG is accumulated in vulnerable plaques due to higher metabolic activity of the inflamed cap and could be used to detect a vulnerable plaque. However, positron emission tomography (PET) cannot detect the FDG-labelled plaques because of respiratory and heart motions, small size and low activity of the plaques. Plaques can be detected using a miniature particle (positron) detector inserted into the artery. In this work, a new detector concept is investigated for intravascular imaging of the plaques. The detector consists of a storage phosphor tip bound to the end of an intravascular catheter. It can be inserted into an artery, absorb the 18F-FDG positrons from the plaques, withdrawn from the artery and read out. Length and diameter of the storage phosphor tip can be matched to the length and the diameter of the artery. Monte Carlo simulations and experimental evaluations of coronary plaque imaging with the proposed detector were performed. It was shown that the sensitivity of the storage phosphor detector to the positrons of 18F-FDG is sufficient to detect coronary plaques with 1 mm and 2 mm sizes and 590 Bq and 1180 Bq activities in the arteries with 2 mm and 3 mm diameters, respectively. An experimental study was performed using plastic tubes with 2 mm diameter filled with an FDG solution, which simulates blood. FDG spots simulating plaques were placed over the surface of the tube. A phosphor tip was inserted into the tube and imaged the plaques. Exposure time was 1 min in all simulations and experiments. Experiments showed that detecting the coronary plaques using the proposed technique is possible. The proposed technique has the potential for fast and accurate detection of vulnerable coronary and other intravascular plaques. PMID- 16467589 TI - Validation of a Monte Carlo simulation of the Philips Allegro/GEMINI PET systems using GATE. AB - A newly developed simulation toolkit, GATE (Geant4 Application for Tomographic Emission), was used to develop a Monte Carlo simulation of a fully three dimensional (3D) clinical PET scanner. The Philips Allegro/GEMINI PET systems were simulated in order to (a) allow a detailed study of the parameters affecting the system's performance under various imaging conditions, (b) study the optimization and quantitative accuracy of emission acquisition protocols for dynamic and static imaging, and (c) further validate the potential of GATE for the simulation of clinical PET systems. A model of the detection system and its geometry was developed. The accuracy of the developed detection model was tested through the comparison of simulated and measured results obtained with the Allegro/GEMINI systems for a number of NEMA NU2-2001 performance protocols including spatial resolution, sensitivity and scatter fraction. In addition, an approximate model of the system's dead time at the level of detected single events and coincidences was developed in an attempt to simulate the count rate related performance characteristics of the scanner. The developed dead-time model was assessed under different imaging conditions using the count rate loss and noise equivalent count rates performance protocols of standard and modified NEMA NU2-2001 (whole body imaging conditions) and NEMA NU2-1994 (brain imaging conditions) comparing simulated with experimental measurements obtained with the Allegro/GEMINI PET systems. Finally, a reconstructed image quality protocol was used to assess the overall performance of the developed model. An agreement of <3% was obtained in scatter fraction, with a difference between 4% and 10% in the true and random coincidence count rates respectively, throughout a range of activity concentrations and under various imaging conditions, resulting in <8% differences between simulated and measured noise equivalent count rates performance. Finally, the image quality validation study revealed a good agreement in signal-to-noise ratio and contrast recovery coefficients for a number of different volume spheres and two different (clinical level based) tumour-to-background ratios. In conclusion, these results support the accurate modelling of the Philips Allegro/GEMINI PET systems using GATE in combination with a dead-time model for the signal flow description, which leads to an agreement of <10% in coincidence count rates under different imaging conditions and clinically relevant activity concentration levels. PMID- 16467591 TI - Diffuse optical tomography through solving a system of quadratic equations: theory and simulations. AB - This paper discusses the iterative solution of the nonlinear problem of optical tomography. In the established forward model-based iterative image reconstruction (MOBIIR) method a linear perturbation equation containing the first derivative of the forward operator is solved to obtain the update vector for the optical properties. In MOBIIR, the perturbation equation is updated by recomputing the first derivative after each update of the optical properties. In the method presented here a nonlinear perturbation equation, containing terms up to the second derivative, is used to iteratively solve for the optical property updates. Through this modification, reconstructions with reasonable contrast recovery and accuracy are obtained without the need for updating the perturbation equation and therefore eliminating the outer iteration of the usual MOBIIR algorithm. To improve the performance of the algorithm the outer iteration is reintroduced in which the perturbation equation is recomputed without re-estimating the derivatives and with only updated computed data. The system of quadratic equations is solved using either a modified conjugate gradient descent scheme or a two-step linearized predictor-corrector scheme. A quick method employing the adjoint of the forward operator is used to estimate the derivatives. By solving the nonlinear perturbation equation it is shown that the iterative scheme is able to recover large contrast variations in absorption coefficient with improved noise tolerance in data. This ability has not been possible so far with linear algorithms. This is demonstrated by presenting results of numerical simulations from objects with inhomogeneous inclusions in absorption coefficient with different contrasts and shapes. PMID- 16467592 TI - A Faraday effect position sensor for interventional magnetic resonance imaging. AB - An optical sensor is presented which determines the position and one degree of orientation within a magnetic resonance tomograph. The sensor utilizes the Faraday effect to measure the local magnetic field, which is modulated by switching additional linear magnetic fields, the gradients. Existing methods for instrument localization during an interventional MR procedure often use electrically conducting structures at the instruments that can heat up excessively during MRI and are thus a significant danger for the patient. The proposed optical Faraday effect position sensor consists of non-magnetic and electrically non-conducting components only so that heating is avoided and the sensor could be applied safely even within the human body. With a non-magnetic prototype set-up, experiments were performed to demonstrate the possibility of measuring both the localization and the orientation in a magnetic resonance tomograph. In a 30 mT m(-1) gradient field, a localization uncertainty of 1.5 cm could be achieved. PMID- 16467593 TI - Determination of heterogeneous thermal parameters using ultrasound induced heating and MR thermal mapping. AB - In this paper, a method for the determination of spatially varying thermal conductivity and perfusion coefficients of tissue is proposed. The temperature evolution in tissue is modelled with the Pennes bioheat equation. The main motivation here is a model-based optimal control for ultrasound surgery, in which the tissue properties are needed when the treatment is planned. The overview of the method is as follows. The same ultrasound transducers, which are eventually used for the treatment, are used to inflict small temperature changes in tissue. This temperature evolution is monitored using MR thermal imaging, and the tissue properties are then estimated on the basis of these measurements. Furthermore, an approach to choose transducer excitations for the determination procedure is also considered. The purpose of this paper is to introduce a method and therefore simulations are used to verify the method. Furthermore, computations are accomplished in a 2D spatial domain. PMID- 16467594 TI - Performance comparison of independent component analysis algorithms for fetal cardiac signal reconstruction: a study on synthetic fMCG data. AB - Independent component analysis (ICA) algorithms have been successfully used for signal extraction tasks in the field of biomedical signal processing. We studied the performances of six algorithms (FastICA, CubICA, JADE, Infomax, TDSEP and MRMI-SIG) for fetal magnetocardiography (fMCG). Synthetic datasets were used to check the quality of the separated components against the original traces. Real fMCG recordings were simulated with linear combinations of typical fMCG source signals: maternal and fetal cardiac activity, ambient noise, maternal respiration, sensor spikes and thermal noise. Clusters of different dimensions (19, 36 and 55 sensors) were prepared to represent different MCG systems. Two types of signal-to-interference ratios (SIR) were measured. The first involves averaging over all estimated components and the second is based solely on the fetal trace. The computation time to reach a minimum of 20 dB SIR was measured for all six algorithms. No significant dependency on gestational age or cluster dimension was observed. Infomax performed poorly when a sub-Gaussian source was included; TDSEP and MRMI-SIG were sensitive to additive noise, whereas FastICA, CubICA and JADE showed the best performances. Of all six methods considered, FastICA had the best overall performance in terms of both separation quality and computation times. PMID- 16467597 TI - Low birth weight and insulin resistance associated with lean body adiposity in an adolescent onset diabetic patient. AB - We describe a patient characterized by reduced fetal growth, no history of rapid "catch-up" growth in prepubertal childhood, and adolescent onset diabetes. High doses of insulin were required for glycemic control. Pioglitazone treatment was followed by a deterioration of glycemic control and hepatic steatosis, while metformin treatment was followed by a partial response. Leptin and adiponectin levels were reduced, but a significant increase in leptin and adiponectin levels occurred concomitantly with an increase in weight and adiposity. This case suggests that some fetal genetic factors determining insulin sensitivity might be linked to a reduction in fetal growth as well as the later development of diabetes. PMID- 16467596 TI - Edaravone diminishes free radicals from circulating neutrophils in patients with ischemic brain attack. AB - OBJECTIVE: Treatment with a free radical scavenger could be a new option for ischemic brain attack, however, little is known about the alteration of oxidative stress markers induced by edaravone, a novel free radical scavenger, in human ischemic brain attack. METHODS: We investigated the effects of edaravone on the oxidative stress markers in patients with ischemic brain attack. Twenty-one patients with ischemic brain attack and 19 controls were enrolled in this study. Blood samples were obtained just before and soon after the first administration of edaravone (30 mg) or ozagrel (40 mg). Intracellular reactive oxygen species of neutrophils were measured using 6-carboxy-2', 7'-dichlorodihydrofluorescin diacetate and a fluorescence-activated cell sorter. Superoxide from neutrophils, induced by phorbol myristate acetate (PMA), was determined by luminol-amplified chemiluminescence assay. RESULTS: Treatment with 30 mg of edaravone significantly decreased the intracellular reactive oxygen species (ROS) of neutrophils (Wilcoxon test, p=0.0001) and PMA-induced superoxide produced by neutrophils (Wilcoxon test, p=0.001). Ozagrel did not alter the intracellular ROS or superoxide production of neutrophils. CONCLUSION: Reduction of intracellular ROS and suppression of superoxide production in neutrophils provide a potential explanation for the clinical efficacy of edaravone in patients with ischemic brain attack. PMID- 16467598 TI - Ventricular fibrillation due to severe hypokalemia induced by steroid treatment in a patient with thyrotoxic periodic paralysis. AB - We report a 25-year-old Japanese man with ventricular fibrillation associated with severe hypokalemia. He developed arm and leg paralysis. He had received 2 g of methylprednisolone because thoracic epidural hematoma had been suspected in another hospital. His serum potassium was 0.8 mEq/l on arrival at our hospital. Half an hour after arrival ventricular fibrillation occurred. Treatment with electric defibrillation 8 times was successful. Afterward Graves' disease was diagnosed, therefore, his clinical symptom was diagnosed as thyrotoxic periodic paralysis. We considered that the unusual condition of hyperthyroid-related hypokalemia worsened by steroid therapy induced the ventricular fibrillation. PMID- 16467599 TI - Pulmonary infection of Mycobacterium avium-intracellulare complex with simultaneous organizing pneumonia. AB - A 67-year-old woman presented high-grade fever and dyspnea. Sputum culture confirmed Mycobacterium avium-intracellulare complex (MAC). Transbronchial lung biopsies revealed organizing pneumonia (OP) that was rapidly improved with corticosteroid. Five months after onset, a nodule emerged in the right lung. Although MAC was confirmed, the lesion was deemed too small to merit anti mycobacterial chemotherapy. Four months later, diffuse infiltrates developed on chest X-ray. Bronchoalveolar lavage study identified MAC and exhibited OP patterns. We commenced antimycobacterial chemotherapy. The infiltrates almost completely improved within a month without corticosteroid. PMID- 16467600 TI - Subcutaneous panniculitis-like T-cell lymphoma: successful initial treatment with prednisolone and cyclosporin A. AB - A case of subcutaneous panniculitis-like T-cell lymphoma (SPTCL) is reported. A 27-year-old man presented with fever and abdominal swelling. His laboratory examination revealed pancytopenia and liver dysfunction. The diagnosis of SPTCL was made by biopsy based on thickened subcutaneous tissue. In addition, bone marrow specimen showed a hemophagocytosis syndrome (HPS). Methylprednisolone pulse therapy was initiated followed by prednisolone (60 mg/day) and cyclosporin A (150 mg/day). He responded to the treatment and remained asymptomatic for at least for 6 months. Our results suggest that a trial of cyclosporin A is warranted in patients with SPTCL complicated by HPS. PMID- 16467601 TI - Venous infarction secondary to septic cavernous sinus thrombosis. AB - A 65-year-old woman with poorly controlled diabetes presented bilateral miosis, bilateral abducens nerve palsy, and left hemiparesis. On MRI, cavernous sinus thrombosis, subdural empyema and hemorrhagic infarction in the frontotemporal lobe were detected. Cerebral angiogram revealed filling defect in the cavernous sinus with venous congestion but no involvement of internal carotid artery. Postmortem examination demonstrated hemorrhagic infarction in the right frontotemporal lobe as well as hemorrhagic necrosis of the pituitary gland. It should be noted that venous congestion due to cavernous sinus thrombosis may cause these complications. PMID- 16467602 TI - "Crazy-paving" appearance in systemic lupus erythematosus. PMID- 16467603 TI - Serologically diagnosed toxocariasis with hemophagocytic syndrome in a patient with primary biliary cirrhosis. PMID- 16467604 TI - X-linked Charcot-Marie-Tooth disease with transient splenium lesion on MRI. PMID- 16467606 TI - Diagnosis of chlamydia-induced reactive arthritis. PMID- 16467605 TI - Intestinal pseudo-obstruction in acute myelitis. PMID- 16467607 TI - [Diabetes mellitus and cellular immunity]. AB - Interaction between diabetes mellitus and cellular immunity is reviewed in several aspects. Importance of cellular immunity in the development of type 1 diabetes mellitus is discussed. Experimental mice models of this disease are presented which after the evaluation of similarities of immunologic parameters provide clues to potential mechanisms of the pathogenesis of type 1 diabetes mellitus. In an animal model of spontaneous type 1 diabetes mellitus, the non obese diabetic mouse develops a spontaneous T cell-mediated insulitis, which is followed by overt diabetes. Disease can be transferred to young non-obese diabetic mice with diabetogenic T cells from older, affected animals. T cells can recognize various islet antigens, including insulin and islet cell enzyme called glutamic acid decarboxylase. Induction of T cell tolerance to these antigens retards the onset of diabetes in mice. Transgenic mouse models of type 1 diabetes mellitus confirm above-mentioned facts. Later alterations of cellular immunity are under the investigation. Disorders of mobilization and chemotaxis, phagocytosis and adherence of polymorphonuclear leukocytes, monocyte function, defects of acquired cellular immunity are thoroughly discussed. Diabetes mellitus, particularly poorly controlled diabetes, impairs different functions and links of cellular immunity. Infections, which are found relatively more frequently in diabetic patients and against which primarily cellular immunity protects, are also reviewed. PMID- 16467608 TI - [Use of strontium-89 in the analgesic treatment of cancer patients with bone metastases]. AB - The aim of this article was to review the available literature regarding to the use of strontium-89 in the palliation of osteoblastic bone pain. The data of many researchers showed that approximately 80% of patients with pain from osteoblastic lesions resulting from prostate or breast cancer experience significant pain relief by administration of strontium-89, with only mild levels of hematotoxicity. The duration of pain relief in some cases exceeded 3-6 months. Indications for administration of strontium-89, effectiveness and duration of the treatment, side effects are reviewed in this article. PMID- 16467609 TI - Influence of catheter on urinary flow during urodynamic pressure-flow study in men with symptomatic benign prostatic hyperplasia. AB - Value of urodynamic pressure-flow studies in evaluation of bladder outlet obstruction has been recognized. Voiding during these studies is influenced by transurethral catheter, which is used for measurement of vesical pressure. We have investigated the influence of 7 F (2.3 mm) catheter on flow rate during pressure-flow study as a potential cause of misclassification. PATIENTS AND METHODS: Data of free urinary flow and pressure flow from 111 men with symptomatic benign prostatic hyperplasia were analyzed. Inclusion criteria for analysis: age over 45 years, total International Prostate Symptom Score over 8, maximum flow rate in range of 4-20 ml/s, total voided volume of 100 ml or greater. RESULTS: Of all patients, means of maximum free and pressure-flow rate were 9.8 and 9.0 ml/s (p=0.01) with mean voided volume 199 and 212 ml (p=0.03) respectively. Maximum flow rate decreased in 56.8%, increased in 41.4% and was stable in 1.8% of cases. The difference ranged from -8.5 to +10.2 ml/s ('+' is indicated when maximum rate of free flow is higher). In the group of obstructed subjects mean maximum flow rates were respectively 8.8 ml/s and 7.9 ml/s (p=0.01). There was no significant difference in maximum flow rate within the group of unobstructed/equivocal subjects. More pronounced mean 1.3 ml/s difference in maximum flow rate was observed also in subgroup of patients with prostate volumes over 60 cc (p=0.01). CONCLUSIONS: Catheter of 7 F (2.3 mm) generally slightly diminishes maximum flow rate. Overdiagnosis of obstruction is more likely if considering the effect of catheter and vesical pressure. Misclassification of subject is possible in case of mild obstruction so such cases should be interpreted with caution. In the case of big difference in maximum flow rate it is necessary to take into account the free flow. PMID- 16467610 TI - The results of the treatment of childhood medulloblastoma with radiotherapy at Kaunas University of Medicine Hospital in 1994-2000. AB - Medulloblastoma, a primitive neuroectodermal tumor growing in cerebellum, is one of the most sensitive to radiation therapy childhood brain tumors, therefore, this method of treatments is justly considered to be the standard for the treatment of medulloblastoma. The outcome of this malignant brain tumor differs in standard and high-risk groups of patients. The aim of the work was to evaluate the survival rate for children with medulloblastoma according to two risk groups. PATIENTS AND METHODS: Eighteen patients aged from 3 to 18 years with histological proven medulloblastoma treated with standard craniospinal and additional posterior fossa radiotherapy were investigated in our study. Nine patients with disseminated and partial removed medulloblastoma were assigned to the high-risk group and other 9 patients with local ant totally removed medulloblastoma were allocated to the standard risk group. RESULTS: Radiological response of medulloblastoma to the radiation therapy was observed in 15 (83.3%) out of 18 patients: complete radiological response was observed in 6 (67%) out of 9 standard-risk patients and in only 1 (11.1%) out of 9 high-risk patients (p<0.05). Medulloblastoma progressed in 15 (83.3%) patients treated with radiation therapy: relapse rate in the high-risk group was 100% and in the standard-risk group--66.7% (p>0.05). The mean time to progression for all patients was 18.2 months: 28.9 months in standard and 7.4 months in high-risk group (p=0.02). The overall survival for all investigated patients was 25.8 months: 37.2 and 14.3 months in the standard and high-risk groups, respectively (p=0.01). Five years progression-free and overall survival rate for all patients was 16.7%: 0% in the high-risk group and 33.3 % in the standard-risk group (p>0.05). CONCLUSION: In our study the difference in survival rate between standard and high-risk patients with medulloblastoma was shown. We observed a statistically significant longer time to progression and better overall survival in the standard-risk group. However, we did not find any significant differences in other survival indices (response, relapse rates, mortality, five years progression- free and overall survival) between those two risk groups. PMID- 16467611 TI - [The role of electroporation in increasing the effectiveness of anticancer drugs]. AB - By applying electric pulses to tumor the membrane of tumor cells undergoes destabilization that enables the drug molecules to enter into the cells more freely and selectivity of anticancer drugs increases. Thus, administering smaller doses of anticancer drugs it might be possible to achieve a better therapeutic effect and to reduce side effects of cytostatic therapy. Such combination of chemotherapy with electric pulses is called electrochemotherapy. The aim of this study was to evaluate the anticancer effectiveness of single and repeated courses of electrochemotherapy with three different anticancer agents (cyclophosphamide, carboplatinum and bleomycin) using experimental animals. It was established that electrochemotherapy used as a single treatment the best increases cytotoxicity of bleomycin. While applying the electrochemotherapy course for 5 days all investigated anticancer drugs showed higher cytotoxicity as compared to the effect of these cytostatics without electroporation. It is notable that in this case antitumor effect of bleomycin was the highest--99% of treated tumors showed a high response rate and no recurrence was observed for 30 days. PMID- 16467612 TI - Effects of parathyroid hormone on newly regenerated bone during distraction osteogenesis in a rabbit tibial lengthening model. A pilot study. AB - OBJECTIVE: The overall purpose of the study is to determine the effects of parathyroid hormone (PTH) (1-34) on bone formation in regenerated and surrounding bone of distracted callus during limb lengthening in rabbits. Additionally the aim of the pilot study is to titrate the optimal dose of PTH for distraction osteogenesis treatment in rabbits' tibial lengthening model. MATERIALS AND METHODS: A total of 18 rabbits underwent right tibia lengthening by callus distraction. Lengthening was started 5 days postoperatively 1 mm/day for a 10-day period and consolidation of 20 days followed. Rabbits were divided into three groups: group I received PTH (1-34) treatment at a dose of 5 microg/kg/day, group II received treatment with PTH (1-34) at a dose of 25 microg/kg/day; group III rabbits were treated with saline. After euthanasia, tibiae of both legs were dissected free, kept frozen and underwent x-ray analysis, dual x-ray absorptiometry-scanning, microcomputed tomography scanning and three-dimensional evaluation and mechanical test followed. RESULTS: Over all, during distraction osteogenesis in a new regenerated bone, PTH (1-34) treatment with two different doses of 5 microg/kg/day and 25 microg/kg/day increased callus cross-sectional area, callus bone mineral density and bone mineral content, bone volume density; dramatically increased trabecular number with slight increase in trabecular thickness, whereas decreased trabecular separation, bone surface density and decreased degree of anisotropy when compared to control group animals. CONCLUSION: PTH (1-34) treatment improved mineralization, structural indices of regenerated distracted rabbits' tibiae, whereas treatment at a dose of 25 microg/kg/day PTH (1-34) was significantly more effective than 5 microg/kg/day PTH(1-34) dose treatment when compared to control group. Bigger dose has been chosen for the main study. PMID- 16467613 TI - The influence of the degree of visual impairment on psychomotor reaction and equilibrium maintenance of adolescents. AB - The changes in such human motor performance processes as stability maintenance, production of a response to the environment may be observed if loss of vision or any vision impairment appears. A total of 45 sighted, legally or totally blind subjects volunteered in the study. The influence of the degree of visual impairment on simple, psychomotor reaction and equilibrium parameters was assessed. Reaction and equilibrium dependence on the degree of visual impairment and the possible existence of compensatory motor reactions of visually impaired adolescents were evaluated in the study. Results allowed thinking that insufficient visual information hampers motor performance and maybe the development of the compensatory motor reactions of the legally blind. Approximate values of the reaction parameters of the sighted with open eyes and the totally blind subjects allow one to predicate that the compensatory motor reactions of the blind probably exist. Testing results of the sighted and totally blind subjects are not equal; therefore, we can suppose that intensified tactile or vestibular function cannot absolutely replace the presence of normal vision in motor control. PMID- 16467614 TI - Anthropometrical data and physical fitness of Lithuanian soldiers according to the sociodemographic characteristics. AB - The aim of this study was to assess the anthropometrical data and physical fitness of soldiers according to the sociodemographic characteristics during one year compulsory military service. The height and weight of 197 soldiers were measured at the beginning and at the end of their service, and body mass index was calculated. The physical fitness was evaluated using index of ability on the basis of 2 km walking test. Conscripts from cities were taller than conscripts from rural areas. At the beginning of the service every fifth was overweight (25< or = body mass index <30 kg/m2) or obese (body mass index > or =30 kg/m2): 17.9% of conscripts from rural areas as well as 17.6% from urban were overweight and 1.9% from rural as well as 1.1% from urban areas were obese. A tendency that the weight of conscripts gradually increased with the level of education was observed. There were no significant differences in index of ability according to the level of soldiers' education as well as place of residence. During one-year service the height of soldiers did not change, however, their weight increased in average by 5.5+/-4.5 kg (p<0.01). The part of overweight persons at the end of the service increased 2.3-fold. Physical fitness of soldiers after one-year service significantly decreased compared with the beginning of the service. In order to decrease the rate of overweight soldiers and to increase their physical fitness the health promotion programs in Lithuanian Army are needed. PMID- 16467615 TI - [Topic on clinical anthropology in scientific activity of docent Antanas Adomaitis]. AB - The present paper is dedicated to docent Antanas Adomaitis, anatomist and anthropologist, former member of editorial board of journal "Medicina", on the occasion of his 65th birthday and 40 years of scientific activity. First publications of Antanas Adomatis appeared in the 1960s, and now the list of his scientific publications exceeds 140 in number. The research field of docent Antanas Adomaitis is wide and covers problems from growth and development to physical status of adult people and its changes associated with a particular period. However, his most important scientific works are in the field of clinical anthropology. Antanas Adomaitis investigated growth and maturation under pathological circumstances, body composition of adult people, prevalence of obesity, growth diversity and secular trend: in 1975 he defended his thesis entitled "Physical development and sexual maturation of children with congenital heart defects" for the degree of Doctor of Medicine, later focused on growth and development of children with congenital facial anomalies, also blind and weak sighted children. Recently docent together with the other scientists of our Department investigates growth variety of children from different towns and rural areas of Lithuania. Antanas Adomaitis collected probably most anthropological data on Lithuanians (children and adults)--this is ponderable contribution to Lithuanian auxology and clinical anthropology, valuable material for the future researches of growth and maturation, also physical status of adult people and secular trend. PMID- 16467616 TI - [The diagnostic value of procalcitonin in severe sepsis]. AB - Sepsis and its complications are the most common cause of the death in the intensive care unit. In spite of the treatment mortality remains up to 28-50%, and 60-90% of the patients are lost because of the complications of sepsis. So it is very important to diagnose this pathology and start the treatment early. The diagnosis of sepsis is complicated for clinical signs and symptoms are not specific and manifest in the patients who have non-infective diseases, when systemic inflammatory response is involved. Parameters of systemic inflammatory response, such as body temperature, heart rate, respiratory rate, leukocyte count, and C-reactive protein concentration, used in clinical practice are neither specific, non sensitive. These parameters often provide information that is inadequate for the discrimination of bacterial and nonbacterial infections and for diagnosis. So it is impossible to differentiate systemic inflammatory response and sepsis. Procalcitonin is a new parameter for diagnosis of bacterial, fungal and parasitical infections. In healthy humans almost all procalcitonin, which is produced in thyroid gland, is resolved and does not reach the blood stream. Its half-life in plasma is only few minutes, so in healthy humans the level of procalcitonin is very low (<0.1 ng/ml) and is not detectable by standard methods. In the case of infection the level of procalcitonin rapidly increases during 2-6 hours and reaches the maximum level after 6-12 hours. The measurement of procalcitonin levels can be used for instant diagnosis as well as for evaluation of the treatment effectiveness. In our article we review the new literature data on the importance of procalcitonin level for sepsis diagnosis in comparison with other parameters of systemic inflammatory reaction, and discuss the indications for procalcitonin analysis. PMID- 16467617 TI - [Acute salicylate poisoning]. AB - Although aspirin (acetylsalicylic acid) has become widely available without prescription, cases of self-poisoning due to overdose of salicylates are quite uncommon, with a low reported mortality. However, severe poisoning with these preparations is life threatening. Besides the aspirin, there are other sources of salicylate poisoning, such as an excessive application of topical agents, ingestion of salicylate containing ointments, use of keratolytic agents or agents containing methyl salicylate (e.g. oil of wintergreen). Most of these preparations are liquid, highly concentrated and lipid soluble, and, therefore, they are able to provoke a severe, rapid salicylate poisoning. On the basis of clinical and metabolic features or salicylate concentration in plasma it is very important to diagnose severe poisoning with salicylates in time and prescribe an adequate treatment. In the present review article various aspects of salicylate poisoning and its treatment are discussed: epidemiology, pharmacokinetics and pharmacodynamics of salicylates, clinical manifestations of their toxicity, management, enhanced elimination and prognosis. PMID- 16467618 TI - Effective bone hemostasis and healing using radiofrequency and conductive fluid. AB - Hemostasis in bone is difficult to achieve because of the mineral content. Current techniques often are ineffective, can have systemic effects, or leave residual material in the wound. Our hypotheses were that a wand device coupling radiofrequency energy with a cooling conductive saline solution, applied topically to bone, could produce superior hemostasis compared with conventional electrocautery or no treatment, and not impede bone healing. Immediate hemostasis and subsequent bone healing for 6 and 12 weeks were evaluated in an iliac crest ostectomy (cancellous bone) and a drilled tibia defect (cortical bone) sheep model. Outcome variables were amount and intensity of bleeding, serial radiography, quantitative computed tomography, histology and mechanical testing. Control of bleeding was nearly complete (93%) and greater with the radiofrequency/saline treatment compared with electrocautery (56%) or no treatment (0%) in cancellous bone and cortical bone. Electrocautery induced surface char (black carbon debris) that could be seen at 6 and 12 weeks. There were no differences in bone healing between the radiofrequency and electrocautery device applications or untreated bone. At 12 weeks, all healing tibiae defects were as strong as undrilled tibiae. This may be an effective method to produce rapid hemostasis in bone without char or healing complications. PMID- 16467619 TI - Case reports: Atlantooccipital and atlantoaxial traumatic dislocation in a child who survived. AB - A 4-year-old child with multilevel spinal injury including atlantooccipital and atlantoaxial dislocation, and thoracic spinal fractures was managed nonoperatively with no significant sequelae from the cervical injuries, but persistent paraplegia from the associated thoracic spinal fractures. Although treatment guidelines cannot be established with the result of one case, we propose an algorithm for treatment of such a rare and severe situation. PMID- 16467620 TI - Acetabular component orientation in 834 total hip arthroplasties using a manual technique. AB - Acetabular component orientation during total hip arthroplasty affects dislocation, range of motion, polyethylene wear, pelvic osteolysis, and component migration. New total hip arthroplasty navigation systems have been introduced to avoid the errors reported after acetabular component orientation using a manual technique. We investigated acetabular component orientation in 834 consecutive primary total hip arthroplasties performed by one surgeon using a manual technique. We examined the relationship between dislocation and a defined safe range. Vertical tilt and planar anteversion were 44.4 +/- 6.5 and 17.9 +/- 6.3 (mean +/- standard deviation), respectively. The error of vertical tilt and planar anteversion were 5.2 +/- 3.9 and 5.3 +/- 4 (mean +/- standard deviation), respectively. Two hundred thirty-three of 834 hips (27.8 %) were outside the safe range. Logistic regression analysis showed that patients with Group IV class according to the classification system of Crowe et al and right hips had a greater risk of being outside the safe range. Navigation systems should be considered, especially in patients with highly dislocated hips. We found no relationship between risk of dislocation and the safe range. Factors other than acetabular component orientation also might affect dislocation. LEVEL OF EVIDENCE: Prognostic Study, Level IV (case series). PMID- 16467621 TI - Outcomes after interposition arthroplasty for treatment of hallux rigidus. AB - Interposition arthroplasty reportedly improves outcomes after traditional salvage procedures for advanced hallux rigidus. We hypothesized this procedure can provide pain relief and satisfactory function with few complications. We examined 18 patients with severe articular cartilage loss who received 21 interposition arthroplasties. The patients a mean age was 56 years. They had a mean followup of 38 months. All patients had substantial loss of articular cartilage when examined intraoperatively. Patients were evaluated using the American Orthopaedic Foot and Ankle Society and Short Form-36 scores. All 18 patients had pain relief, and 17 of 18 patients said they would have the procedure again. The mean postoperative increase in range of motion of the first meta-tarsophalangeal joint was 37 degrees . The mean American Orthopaedic Foot and Ankle Society and Short Form-36 scores were 78.4 and 96.3, respectively. The complication rate was 6%. Results of our study indicate that interposition arthroplasty relieves pain and restores motion in patients with advanced hallux rigidus and may offer a reliable option to fusing the joint. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series). PMID- 16467622 TI - Clavicula pro humero reconstruction after wide resection of the proximal humerus. AB - The optimal reconstruction procedure after resection of proximal humeral bone tumors is controversial. We performed clavicula pro humero reconstruction in seven consecutive patients after wide resection of malignant proximal humerus tumors to determine the functional outcomes and to clarify whether this procedure could reduce the occurrence of complications. The histologic diagnoses were conventional osteosarcoma in four patients, chondrosarcoma in two, and parosteal osteosarcoma in one. Postoperative function was assessed with the Musculoskeletal Tumor Society upper extremities scale and an evaluation chart for activities of daily living. The mean Musculoskeletal Tumor Society score was 69%. Stable results were obtained regardless of the extent of the resection, and the results as a flail shoulder were satisfactory. The mean daily activity rating was approximately 80%. The activities of daily living functional rating was better (92%) when the nondominant hand was affected compared with the dominant hand (69%). Bony union was achieved in all seven patients. We observed no instances of infection, fracture, or nonunion. The clavicula pro humero reconstruction is a useful procedure that could reduce the occurrence of complications, especially in young patients and in patients with involvement of the nondominant arm. PMID- 16467624 TI - Intraoperative electron radiotherapy for extremity sarcomas does not increase acute or late morbidity. AB - Intraoperative electron radiotherapy is used to treat surgical sites that potentially harbor occult tumor immediately after limb-sparing surgical resection of extremity soft tissue sarcomas. It is unknown whether single-fraction, high dose intraoperative electron radiotherapy at the time of surgery increases wound morbidity when combined with preoperative or postoperative external beam radiotherapy. In a retrospective study, we evaluated whether intraoperative electron radiotherapy increased 90-day and late (> 90 days) wound complication rates by comparing patients who had adult extremity soft tissue sarcomas treated by limb-sparing surgery and preoperative (n = 14) or postoperative (n = 13) external beam radiotherapy. The median followup was 36 months. Seven (26%) patients had wound complications occurring within 90 days postoperatively and completion of radiotherapy. Late wound complication rates were similar. Two patients in each of the external beam radiotherapy groups required late subtotal limb amputations for prolonged wound complications. Our findings suggest intraoperative electron radiotherapy during limb-sparing surgery allows radiation dose escalation without increased 90-day or late-wound complication rates when combined with preoperative or postoperative external beam radiotherapy for patients with extremity soft tissue sarcomas. LEVEL OF EVIDENCE: Prognostic Study, Level II (retrospective study). See the Guidelines for Authors for a complete description of levels of evidence. PMID- 16467623 TI - Osseointegration of cementless implants with different bisphosphonate regimens. AB - Some evidence suggests a daily dose of bisphosphonates improves fixation of cementless metal implants by enhancing osseointegration. Because the necessity of daily administration may result in suboptimal adherence to therapy, single- dose administration is desirable. We examined whether a dose-equivalent single injection of the nitrogen-containing bisphosphonate ibandronate is as effective as a daily injection in improving the osseointegrated surface and enhancing periprosthetic bone mineralization (bone volume to tissue volume) of cementless metal implants. Uncoated titanium and hydroxyapatite-coated titanium implants were surgically inserted into the femoral medullary canal of 55 female Sprague Dawley rats. The animals were randomly assigned subcutaneous treatments with 25 microg/kg body weight ibandronate per day, a dose-equivalent single injection of 28 x 25 microg/kg body weight, or saline solution for control. Histomorphometric evaluation revealed an enhanced osseointegrated surface for hydroxyapatite-coated implants in both treatment groups, but only for daily injections for uncoated titanium implants. Bone volume to tissue volume was improved in both treatment groups. Our results suggest that an equivalent-dose single injection of ibandronate is as effective as a daily dose in improving osseointegration and stabilization of hydroxyapatite-coated titanium implants in this rat model. PMID- 16467625 TI - A laterally positioned concave trochlear groove prevents patellar dislocation. AB - Patellofemoral instability is a disabling condition that occurs in adolescence. Recurrence after patellar dislocation has been reported in 2-50% of patients. We compared the distal femur in patients with patellofemoral instability with distal femura of a healthy cohort using mathematical quantification of two-dimensional shape variation at the same position in different knees. One hundred eight computed tomography scans from 54 patients with patellofemoral instability were compared with 197 computed tomography scans from 102 subjects with normal knees. We used principal components analysis to quantify variation in shape of the trochlear groove as it allows comparison of knees with patellofemoral instability to healthy knees. We found that subjects who had dislocated their patella had a flattened trochlear groove, whereas normal knees had a more concave groove. The position of the trochlear groove was more medial in patients with patellofemoral instability compared with normal knees. Groove position was more important than shape when predicting which patellae were likely to dislocate. PMID- 16467626 TI - Fixation of posterior malleolar fractures provides greater syndesmotic stability. AB - Syndesmotic injuries are common in ankle fractures. Traditional syndesmosis fixation may be associated with a secondary procedure. When the posterior malleolus is fractured, the posterior syndesmotic ligaments may remain intact and attached to the fragment. Our goals were to establish the incidence of syndesmotic ligament ruptures in pronation-external rotation type ankle injuries associated with posterior malleolar fractures, and to assess syndesmotic stability after fixation of the posterior malleolus compared with using a syndesmotic screw. Fifteen patients who sustained pronation-external rotation Stage 4 ankle fractures that involved the posterior malleolus were evaluated using radiographs and magnetic resonance imaging. No complete tears of the posterior-inferior tibiofibular ligament occurred. A pronation-external rotation fracture pattern with a posterior malleolar fragment was created in 10 lower extremity cadaver specimens with random fixation of the posterior malleolus or the syndesmosis. Compared with the intact specimens, stiffness was restored to 70% after fixation of the posterior malleolus, and to 40% after syndesmosis stabilization. Syndesmotic stability may be obtained more effectively by fixation of the posterior malleolus rather than by using a syndesmotic screw. Although additional clinical investigation is warranted, these concepts may be useful in eliminating syndesmotic screw fixation in select patients. PMID- 16467627 TI - Case reports: long thoracic nerve palsy after using a single axillary crutch. AB - We describe a 42-year-old patient who incurred long thoracic nerve palsy after using one axillary crutch. The patient sustained fractures to his left tibia and fibula. The fractures required internal fixation with a locked intramedullary nail and postoperative restricted weightbearing using axillary crutches. When weightbearing was progressed, the patient began using one axillary crutch on the right side. The resulting asymmetric gait pattern forced the crutch onto his chest wall, which led to right-sided long thoracic nerve palsy. The patient had not recovered 6 months after the injury. When providing axillary crutches, clinicians must consider potential compromise of the long thoracic nerve and the brachial plexus if the crutch is used inappropriately. PMID- 16467628 TI - Arthroscopic and open Bankart repairs provide similar outcomes. AB - Recurrent instability after arthroscopic Bankart repair has decreased, largely because of improvements in surgical technique. We wanted to know whether there were differences in functional outcomes (using a validated outcomes measure) in patients who had arthroscopic Bankart repair or open Bankart repair for recurrent anterior glenohumeral instability. We retrospectively reviewed 106 patients who had a Bankart repair for recurrent anterior glenohumeral instability from 1998 2001. Of the 93 patients included, 69 patients had arthroscopic Bankart repair and 24 patients had open Bankart repair. The indications for surgery in the two groups were similar. The average age of the patients was 29.9 years (arthroscopic Bankart repair, 31 years; open Bankart repair, 28 years). The followup ranged from 24-77 months. The average modified American Shoulder and Elbow Surgeons score (PENN score) for pain, satisfaction, and function were 26.3, 8.5, and 55.1, respectively, in the arthroscopic Bankart repair group and 26.6, 8.8, and 54.2, respectively, in the open Bankart repair group. The total score was 90 in the arthroscopic Bankart repair group and 89.5 in the open Bankart repair group. Recurrent instability occurred in one patient in each group. We found no difference in outcomes between the arthroscopic and open Bankart repair groups using patient-assessed outcomes. LEVEL OF EVIDENCE: Therapeutic Study, Level III (retrospective comparative study). See the Guidelines for Authors for a complete description of levels of evidence. PMID- 16467630 TI - Endoscopic surgery for juvenile nasopharyngeal angiofibroma: where are the limits? AB - PURPOSE OF REVIEW: This article will examine recent publications on the endoscopic management of juvenile nasopharyngeal angiofibromas. RECENT FINDINGS: The use of an endoscopic approach to resect small juvenile nasopharyngeal angiofibromas is supported by excellent results from a number of operative series published in recent years. Large juvenile nasopharyngeal angiofibromas continue to present a considerable surgical challenge, with most being resected by traditional open approaches. However, the results achieved after the endoscopic resection of large tumours have been reported for a small number of cases. This review focuses on the outcome of these cases, which would seem to compare favourably to open series, and the advances in instrumentation and techniques, that facilitate the endoscopic removal of large juvenile nasopharyngeal angiofibromas. SUMMARY: Improvements in preoperative assessment and preparation, operative technique, and instrumentation potentially enable the endoscopic removal of most juvenile nasopharyngeal angiofibromas. PMID- 16467631 TI - A review of the endoscopic approach to the pituitary through the sphenoid sinus. AB - PURPOSE OF REVIEW: The refinement of minimally invasive endoscopic techniques has resulted in 'pure' endoscopic endonasal trans-sphenoidal surgery, which is a new approach for the removal of pituitary tumors. RECENT DEVELOPMENTS: This procedure is performed via a wide anterior sphenoidotomy with detachment of the septum from the sphenoid face, and avoids the use of a trans-sphenoidal retractor and any intraoral or nasal incisions. Straight and angled endoscopes are used throughout the procedure to provide a wide view of the sella and are manipulated by a co surgeon. This technique represents an improvement over pituitary microsurgery, with decreased post-operative morbidities and a shortened postoperative stay, and it eliminates the need for packing while providing an opportunity to monitor the sella after surgery. The technique has been established as being efficacious and safe. It incorporates image-guided surgery, with the fusion of computer tomography and magnetic resonance imaging, and employs new and dedicated instrumentation. Training in endoscopic techniques is required. SUMMARY: Future advancements in intraoperative imaging, cranial base reconstruction, and robotics will make this technique even more successful. PMID- 16467632 TI - Endoscopic management of inverted papilloma. AB - PURPOSE OF REVIEW: Inverted papillomas are uncommon benign tumors that may occur in the sinonasal tract. Because of their tendency towards recurrence and their potential for malignant transformation, complete en bloc resection has been the standard treatment modality. With the application of minimally invasive surgical techniques in the management of sinonasal disease processes, transnasal endoscopic removal of inverted papillomas is gaining in acceptance. The following represents a review of the current literature regarding sinonasal inverted papilloma and its clinical management. RECENT FINDINGS: The recent literature has further refined the technique of endoscopic resection of inverted papillomas by delineating essential principles and applying new technologies, such as image guidance and angled endoscopic drills. Modified approaches and methodology have been described to address tumors originating in particular anatomic locations. At the same time, new information relevant to the diagnosis and surveillance of inverted papillomas has been published, regarding the role of imaging, serum tumor markers, and human papillomavirus. SUMMARY: Growth in experience with the endoscopic management of inverted papillomas has led to evolving technical improvements and innovations. In most tumor locations, the completeness of resection achieved by a skilled endoscopic surgeon is equivalent, if not superior, to that attained with an open approach. In some instances, an endoscopic exploration with tumor resection may help define the site of tumor attachment and direct adjunctive open procedures when indicated. Long-term outcomes studies with sizeable patient cohorts will be needed to define the role of various surgical strategies in the optimal management of inverted papilloma. PMID- 16467633 TI - Staphylococcus aureus and endoscopic sinus surgery. AB - PURPOSE OF REVIEW: The current recommendation for the choice of empiric antibiotic therapy of chronic rhinosinusitis is based on the bacteriology of acute rhinosinusitis. Staphylococcus aureus has been found consistently in cultures from patients with chronic rhinosinusitis. New studies have examined the role of Staphylococcus aureus in the pathogenesis and clinical course of chronic rhinosinusitis, both before and after surgical therapy. The consequence of antibiotic resistance, on the course of chronic rhinosinusitis and its treatment, has also been examined. RECENT FINDINGS: Several recent studies have confirmed past literature demonstrating a high incidence of sinus cultures positive for Staphylococcus aureus growth in patients with chronic rhinosinusitis. Moreover, one study reported that Staphylococcus aureus is common in patients with persistent chronic rhinosinusitis after endoscopic sinus surgery. These studies, however, have not shown that this bacterium is a significant factor in the development of chronic rhinosinusitis or a predictor of the severity of symptoms. Other studies did not support the notion of increased antibiotic resistance in chronic rhinosinusitis, although there is some evidence that antibiotic resistance may lead to refractory cases of the disease. The standard treatment, of multiple courses of empiric antibiotics followed by surgery, was not shown to increase the prevalence of methicillin-resistant Staphylococcus aureus. SUMMARY: A review of recent literature shows a high incidence of positive cultures for Staphylococcus aureus from the sinuses of patients with chronic rhinosinusitis, both before and after surgery, but has not produced convincing evidence that Staphylococcus aureus has a significant role in the pathogenesis or clinical course of the disease. PMID- 16467634 TI - An updated review of clinical olfaction. AB - PURPOSE OF REVIEW: Disorders of the sense of smell can result through hundreds of different processes, but most commonly occur from upper-respiratory-tract infections, trauma, and chronic rhinosinusitis. RECENT DEVELOPMENTS: Research in the basic science of olfaction has progressed rapidly with powerful new molecular discoveries; however, our ability to treat these disorders remains limited. In clinical olfaction we are just realizing the broader existence of the sensory dysfunction in our population. We are discovering associations between neurodegenerative disorders and smell function that may allow us to identify these disorders earlier in the disease process. We are also challenging our previous categorization schemes and realizing that many etiologies cross the traditional conductive and neuro-sensory divisions. SUMMARY: Currently, aside from the possible therapeutic potential of systemic steroids, we have no effective treatment for the most common causes of olfactory loss. Recent advances in the basic science of olfaction provides us with an opportunity to develop new and novel clinical studies in an attempt at improving the quality of life for many of these patients. PMID- 16467635 TI - Acoustic rhinometry: should we be using it? AB - PURPOSE OF THE REVIEW: The purpose of this review is to examine the role of acoustic rhinometry in clinical practice. Although acoustic rhinometry was first described for clinical use in 1989, it is not in common use today. Should we be using it? Yes. I think we should be using it more often. This review provides an update of the new standard for interpretation and expanded clinical uses. RECENT FINDINGS: The most significant advances in the past year in this area have been the publication of standards for its clinical use. In addition, the repertoire of clinical problems that can be analyzed objectively with acoustic rhinometry has expanded to include turbinoplasty, sleep disorders, more types of cosmetic/reconstructive procedures, sinus surgery, vasomotor rhinitis, maxillofacial expansion procedures, and aspirin and methacholine challenge. (Its ability for pediatric disorders, such as adenoidectomy, has been reaffirmed.) Some case examples are included to demonstrate the utility of acoustic rhinometry for 'mixed' pathology. SUMMARY: Acoustic rhinometry is a rapid, objective, painless, noninvasive technique for assessing nasal airway obstruction. Recently, standards have been developed that aid its expansion for clinical use. Expanded clinical applications include sleep disorders, cosmetic/reconstructive and maxillofacial disorders, sinus and turbinate procedures, and pediatrics. Acoustic rhinometry should be utilized to improve our ability to practice evidence-based medicine in rhinology. PMID- 16467636 TI - The hypoplastic maxillary sinus and the orbital floor. AB - PURPOSE OF REVIEW: Hypoplastic maxillary sinus is a relatively rare clinical problem that has a variable presentation. The purpose of this article is to review the recent literature with regard to evaluation and treatment. RECENT FINDINGS: Most authors suggest that hypoplastic maxillary sinus results from the development of negative pressure resulting from an obstruction of maxillary sinus ventilation. Patients most frequently present with unilateral enophthalmos but some cases may be identified incidentally on imaging studies. SUMMARY: Hypoplastic maxillary sinus is a rare clinical entity with variable presentation. Evaluation and management are tailored to each individual patient's degree of disease and symptoms. PMID- 16467638 TI - Bibliography. Current world literature. Nose and paranasal sinuses. PMID- 16467637 TI - Should otolaryngologists perform pH probe studies? AB - PURPOSE OF REVIEW: This commentary discusses recent developments in the methods available to pediatric otolaryngologists in the diagnosis of laryngopharyngeal reflux. RECENT DEVELOPMENTS: The development of detachable wireless monitoring allows the otolaryngologists to perform their own pH monitoring. It can be deployed while performing other procedures, such as laryngoscopy and bronchoscopy. The procedure to insert the probe and collect data is described. SUMMARY: It is hoped that the pH probe will play an important role in understanding the pathological role of pharyngeal reflux in terms of laryngeal, pharyngeal, sinus and ear pathology. PMID- 16467639 TI - Blood pressure and the global burden of disease 2000. Part 1: estimates of blood pressure levels. AB - OBJECTIVE: To provide global estimates of blood pressure by age and sex for adults aged > or = 30 years, by WHO subregion. DESIGN AND METHODS: Data were obtained from studies identified in a literature review of population-based surveys. These were complemented by data from MONICA and INTERSALT studies. Estimates of the shape of the age-systolic blood pressure (SBP) association were made from survey data utilizing parametric and non-parametric analyses. A linear sex-specific association of SBP with age was demonstrated from 30 to 70 years in females and 20 to 70 years for males in each subregion. Mean age- and sex specific estimates of SBP were estimated for each WHO subregion separately, based on study and country-weighted SBP data. RESULTS: Analyses were based on data from about 230 surveys and over 660 000 participants. Age-specific mean SBP values ranged from 114 to 164 mmHg for females, and 117-153 mmHg for males. Females typically had lower SBP levels than males in the 30-44-year age groups, but in all subregions, SBP levels rose more steeply with age for females than males. Therefore, SBP levels in those aged > or = 60 years tended to be higher in females. Subregions with consistently high mean SBP levels included parts of eastern Europe and Africa. Mean SBP levels were lowest in south-east Asia and parts of the western Pacific. CONCLUSIONS: These global estimates of blood pressure by age, sex and subregion show considerable variation in estimated levels. The lack of data in developing countries is substantial, and this is an important limitation given the role of blood pressure in increasing cardiovascular disease levels. PMID- 16467640 TI - Blood pressure and the global burden of disease 2000. Part II: estimates of attributable burden. AB - OBJECTIVES: To provide estimates of the global burden of disease attributable to non-optimal blood pressure by age and sex for adults aged > or = 30 years, by WHO subregion. METHODS: Estimates of attributable burden were made using population impact fractions, which used data on mean systolic blood pressure levels, disease burden [in deaths and/or disability-adjusted life years (DALYs)] and relative risk corrected for regression dilution bias. Estimates were made of burden attributable to a population distribution of blood pressure with a mean systolic blood pressure of greater than 115 mmHg. RESULTS: Globally, approximately two thirds of stroke and one-half of ischaemic heart disease were attributable to non optimal blood pressure. These proportions were highest in the more developed parts of the world. Worldwide, 7.1 million deaths (approximately 12.8% of the global total) and 64.3 million DALYs (4.4% of the global total) were estimated to be due to non-optimal blood pressure. Overall approximately, two-thirds of the attributable burden of disease occurred in the developing world, approximately two-thirds in the middle age groups (45-69 years) and approximately one-half occurred in those with systolic blood pressure levels between 130 and 150 mmHg. CONCLUSIONS: The burden of non-optimal blood pressure is almost double that of the only previous global estimates, which is largely explained by the correction for regression dilution adopted in these analyses. High blood pressure is a leading cause of global burden of disease, and most of it occurs in the developing world. PMID- 16467641 TI - The lower the better: does simplicity lead to absurdity? PMID- 16467642 TI - Cardiovascular risk of systolic versus diastolic blood pressure in Western and non-Western countries. PMID- 16467643 TI - Isolated clinic hypertension in a general population. PMID- 16467644 TI - Can mutations in ion channel genes cause endocrine dysfunction? PMID- 16467645 TI - Candidate gene studies: accepting negative results. PMID- 16467646 TI - New insights into urinary proteins as markers of cardiovascular risk in hypertension. PMID- 16467647 TI - Blood pressure change in a free-living population-based dietary modification study in Japan. AB - OBJECTIVE: To assess whether dietary intervention in free-living healthy subjects is effective in improving blood pressure levels. DESIGN: Open randomised, controlled trial. SETTING: Free-living healthy subjects in two rural villages in north-eastern Japan. PARTICIPANTS: Five hundred and fifty healthy volunteers aged 40-69 years. INTERVENTIONS: Tailored dietary education to encourage a decrease in sodium intake and an increase in the intake of vitamin C and carotene, and of fruit and vegetables. MAIN OUTCOME MEASURES: Blood pressure, dietary intake and urinary excretion of sodium, dietary carotene and vitamin C, and fruit and vegetable intake data were collected at 1 year after the start of the intervention. RESULTS: During the first year, changes differed significantly between the intervention and control groups for dietary (P = 0.002) and urinary excretion (P < 0.001) of sodium and dietary vitamin C and carotene (P = 0.003). Systolic blood pressure decreased from 127.9 to 125.2 mmHg (2.7 mmHg decrease; 95% confidence interval, -4.6 to -0.8) in the intervention group, whereas it increased from 128.0 to 128.5 mmHg (0.5 increase; -1.3 to 2.3) in the control group. This change was statistically significant (P = 0.007). In contrast, the change in diastolic blood pressure did not significantly differ between the groups. In hypertensive subjects, a significant difference in systolic blood pressure reduction was seen between the groups (P = 0.032). CONCLUSION: Moderate intensity dietary counseling in free-living healthy subjects achieved significant dietary changes, which resulted in a significant decrease in systolic blood pressure. PMID- 16467648 TI - Age-specific effects of systolic and diastolic blood pressures on mortality due to cardiovascular diseases among Japanese men (NIPPON DATA80). AB - OBJECTIVE: To clarify the relationship between mortality due to cardiovascular diseases (CVD) and systolic blood pressure (SBP) and diastolic blood pressure (DBP) in young and elderly Japanese men in the absence of antihypertensive treatments. DESIGN: A 19-year cohort study using the 1980 National Survey of Cardiovascular Diseases and the identification of the underlying causes of death using national vital statistics data. METHODS: In 1999, a 19-year follow-up study was conducted among participants randomly selected from the Japanese population for the 1980 National Survey of Cardiovascular Diseases. Among the male participants that were not receiving antihypertensive treatment, information was obtained for about 91.3% of the subjects (n = 3779). Age-adjusted mortality rates using the person-year method were calculated to compare the relationships of SBP and DBP levels with CVD mortality for different age groups (aged 30-64, 65-74, and 75 years or more). RESULTS: The total number of observed person-years was 64 598 and the number of CVD-related deaths after 19 years was 253. Both SBP and DBP levels were significantly and linearly related to CVD mortality in the age groups 30-64 years (P < 0.001 and P = 0.01, respectively) and 65-74 years (P < 0.001 and P = 0.03, respectively). In the group of participants that were at least 75 years old, however, no significant increase in the relative risk of CVD was observed with increasing DBP levels (P = 0.156), whereas the relative risk of CVD increased significantly with increasing SBP levels (P = 0.038). Multivariate analysis adjusting for major risk factors confirmed these relationships. Standardized hazard ratios (95% confidence intervals) of SBP were 1.53 (1.19 1.96) for 30-64 year olds, 1.70 (1.31-2.20) for 64-74 year olds, and 1.23 (1.03 1.47) for participants 75 years old or more, whereas those of DBP were 1.52 (1.12 2.06), 1.60 (1.21-2.10), and 1.10 (0.94-1.28), respectively. CONCLUSIONS: These data show that elevated SBP is an independent risk factor for CVD mortality for Japanese men of all ages, whereas elevated DBP is not an independent risk factor for CVD mortality for elderly men. PMID- 16467649 TI - Prevalence and determinants of isolated clinic hypertension in the Finnish population: the Finn-HOME study. AB - BACKGROUND: Previous studies performed in selected hypertensive subjects have reported several possible determinants of isolated clinic hypertension (ICH). The purpose of this study was to assess the prevalence and determinants of ICH in a randomly selected nationwide population. METHODS: We studied a representative sample of the general adult population (1440 45-74-year-old subjects) in Finland not treated for hypertension. The subjects were drawn from the participants of a multidisciplinary epidemiological survey, the Health 2000 Study. Subjects included in the study underwent a clinical interview, determination of serum lipids and glucose, measurement of clinic and home blood pressure (BP), and psychometric tests for psychological distress, hypochondriasis, depression, and alexithymia. The diagnosis of ICH was based on a clinic BP of 140/90 mmHg or greater and a home BP less than 135/85 mmHg. RESULTS: The prevalence of ICH in the untreated Finnish adult population was 15.6 and 37.5% among untreated clinic hypertensive individuals. In a multivariate logistic regression analysis, ICH was associated with mildly elevated systolic and diastolic BP, lower body mass index (BMI), and non-smoking status. Subjects with ICH represent an intermediate group between the normotensive and sustained hypertensive individuals where cardiovascular risk is concerned (age, BP, diabetes prevalence, lipid profile, and BMI). CONCLUSION: ICH is a common phenomenon in the general population. Non smoking individuals with mildly elevated BP and low BMI have a higher risk of ICH. Physicians should disassociate the diagnosis of ICH from any psychosocial disorders, but should remember that patients with ICH have an increased risk of cardiovascular disease. PMID- 16467650 TI - Haplotypes of the beta-2 adrenergic receptor associate with high diastolic blood pressure in the Caerphilly prospective study. AB - OBJECTIVES: Current evidence demonstrates that both genetic and environmental factors influence blood pressure. The sympathetic nervous system is a key player in blood pressure control and functional genetic variants of the beta-2 adrenergic receptor (B2AR) have been identified and implicated in the pathogenesis of hypertension. The present study aimed to determine the effects of common haplotypes of the B2AR gene upon blood pressure in the Caerphilly Prospective Study. DESIGN: Two thousand five hundred and twelve men (aged 45-59 years) participated in the study. We selected individuals in the upper (n = 347) and lower (n = 279) quintiles of the diastolic blood pressure distribution fixed at two time points [phase 2 (1984-88) or phase 3 (1989-93)] as cases and controls. METHODS: We analysed two functional polymorphisms (Arg16Gly and Gln27Glu) of B2AR and their haplotypes. RESULTS: We found a higher risk of hypertension in individuals homozygous for the Gln27 compared to those individuals homozygous for Glu27 [odds ratio (OR) = 1.94; 95% confidence interval (CI) = 1.34-2.81; P = 0.001]. Three haplotypes (Gly16Gln27, Gly16Glu27 and Arg16Gln27) were present in both quintile groups. Logistic regression analysis showed that haplotypes with a Gln27 allele (Gly16Gln27 and Arg16Gln27) conferred a significantly higher risk for hypertension than the Gly16Glu27 haplotype (OR = 1.55; 95% CI = 1.11-2.17, OR = 1.37; 95% CI = 1.04-1.81; P = 0.009 and P = 0.027, respectively). However, there was no evidence to support a statistically significant difference in odds ratios for the Gly16Gln27 and Arg16Gln27 haplotypes (P = 0.477), suggesting that it is the Gln27 allele alone, rather than any haplotype, which best explains the association. CONCLUSIONS: In a prospectively studied Caucasian male cohort, high diastolic blood pressure was associated with B2AR haplotypes containing the pro-downregulatory Gln27 variant. PMID- 16467651 TI - The functional HERG variant 897T is associated with Conn's adenoma. AB - OBJECTIVE: Aldosterone secreting adenomas (aldosteronomas) have an unknown molecular origin. Ion channel currents are involved in signal transduction leading to aldosterone synthesis and secretion. HERG (human-ether-a-go-go-related gene) encodes for a potassium channel responsible for the outward rectifying delayed current and it is mutation prone. When mutated it causes most of the familial forms of both long QT and short QT syndromes. Abnormal repolarization in glomerulosa cells might increase aldosterone secretion or induce a proliferative advantage. The aims of this study were to: (1) evaluate HERG expression in aldosteronomas; (2) search for HERG somatic mutations; and (3) determine whether there is any relationship between the common HERG functional variant (A2690C, leading from lysine 897 to threonine, K897T) and aldosteronoma. DESIGN AND METHODS: Aldosteronoma and blood samples from 17 patients were studied to evaluate HERG expression, full-length HERG complementary DNA sequencing, and genotyping for K897T alleles. The prevalence of HERG 897 alleles was also tested in a control population and a population consisting entirely of hypertensive individuals. RESULTS: HERG was expressed in all aldosteronomas analysed. HERG somatic mutations were not detected. The 897T variant of HERG was significantly more common among patients with aldosteronoma (897T allele 41%) than in patients with moderate-severe essential hypertension (897T allele 20%, P = 0.007) or in the control population (897T allele 12%, P < 0.0001). The 897T/T genotype was present in 24% of the aldosteronoma patients versus 7% (P = 0.040) and 3% (P = 0.001) in essential hypertension and in the control population, respectively. When the chi test was performed considering the three groups together, the significance was similar (for alleles P < 0.0001 and for genotypes P = 0.004). CONCLUSION: The common functional HERG variant 897T may predispose to the development of aldosteronoma. PMID- 16467652 TI - Effect of vasoconstrictor coupling factor 6 on gene expression profile in human vascular endothelial cells: enhanced release of asymmetric dimethylarginine. AB - BACKGROUND: Coupling factor 6 (CF6), a component of ATP synthase, inhibits phospholipase A2 and induces vasoconstriction. However, because arachidonic acid acts in the widespread fields of vascular biology, CF6 might exert profound effects in addition to vasoconstriction. We investigated the effect of CF6 on the gene expression profile in human umbilical vein endothelial cells. METHODS AND RESULTS: The increased gene expression after 24-h exposure to CF6 at 10 mol/l, assessed by cDNA microarray (n = 3), included neuregulin-1 (1.84 +/- 0.07 fold compared with control, P < 0.05) and relaxin-1 (1.74 +/- 0.20, P < 0.05), both relating to congestive heart failure, urokinase type plasminogen activator receptor (1.77 +/- 0.24, P = 0.06) and estrogen receptor beta (1.74 +/- 0.36, P = 0.08), both relating to vascular inflammation and cell infiltration, and protein arginine methyltransferase (PRMT-1; 1.73 +/- 0.20, P < 0.05). Out of these genes, the enzyme relating to the synthesis (PRMT-1) of asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase (NOS), was further examined concomitantly with the degradation enzyme, dimethylarginine dimethylaminohydrolase 2 (DDAH-2). The ratio of PRMT-1 to glyceraldehyde 3 phosphate dehydrogenase (GAPDH) mRNA, measured by real-time quantitative reverse transcription-polymerase chain reaction, was increased by 9 +/- 2% (n = 10, P < 0.01) at 48 h after CF6 at 10 mol/l, whereas the ratio of DDAH-2 to GAPDH was decreased by 12 +/- 2% (n = 8, P < 0.01). DDAH-2 protein and activity were decreased by 28 +/- 5% (n = 5, P < 0.01) and 19 +/- 2% (n = 6, P < 0.01) by CF6, respectively. ADMA release was enhanced by 20 +/- 8% and NOS activity was decreased by 13 +/- 1% (both n = 8, P < 0.05) by CF6. CONCLUSIONS: CF6 changes the gene expression profile to be proatherogenic and functions as a novel stimulator for ADMA release by enhancing its synthesis and suppressing its degradation. PMID- 16467653 TI - Genetic variation in fibrillin-1 gene is not associated with arterial stiffness in apparently healthy individuals. AB - OBJECTIVE: Arterial stiffness is an independent determinant of cardiovascular risk, and there is evidence that it has a strong genetic component. Fibrillin-1 (FBN-1) is the disease gene for Marfan's syndrome and an FBN-1 polymorphism has been associated with large artery stiffening and elevated pulse pressure (PP) in patients with cardiovascular disease. The aim of this study was to investigate the possible influence of the common FBN-1 genotypes on arterial stiffness in a large cohort of healthy individuals. SUBJECTS AND METHODS: A total of 742 individuals free from cardiovascular disease or risk factors were studied. Aortic pulse wave velocity (PWV) and augmentation index (AIx), blood pressure, lipids and glucose were assessed. Genomic DNA was extracted, and genotyping for the FBN 1 variable nucleotide tandem repeat (VNTR) was performed using a CEQ 8000 sequencer. RESULTS: The mean age (+/- SEM) of the cohort was 49 +/- 1 years. The three common VNTR genotypes accounted for 87.1% of the population frequency. Their frequencies were: 52.3%, 2-2; 16.3%, 2-3; 18.5% 2-4. There were no significant differences in the blood pressure, AIx, PWV, lipids or body mass index among the common genotypes. Moreover, the FBN-1 genotype was not associated with either aortic PWV or other measures of stiffness after correction for other confounding factors. CONCLUSION: These data do not support the hypothesis that aortic PWV or PP are influenced by the FBN-1 VNTR genotype. Although we cannot exclude small effects, this negative finding also suggests that there is not a major allele for stiffness or blood pressure in apparently healthy individuals linked to this VNTR. PMID- 16467654 TI - Turbulent flow/low wall shear stress and stretch differentially affect aorta remodeling in rats. AB - OBJECTIVE: The present investigation was carried out to evaluate the relationship between local hemodynamic forces and intimal and medial remodeling in the proximal and distal segments of the arterial walls of rats in relation to severe infradiaphragmatic stenosis of the aorta. METHODS: Young male rats were divided randomly into an operated group, animals submitted to surgical abdominal aorta stenosis, and a sham-operated group, a control group of animals submitted to sham operation to simulate abdominal aorta stenosis. RESULTS AND CONCLUSIONS: Constricted aortas showed two distinct adaptive remodeling responses to hemodynamic stimuli induced by infradiaphragmatic coarctation. The first is remodeling in the hypertensive prestenotic segment with increased circumferential wall tension (CWT), associated with normal tensile stress, laminar flow/normal wall shear stress characterized by enlarged heterogeneous endothelial cells, elongated in the direction of the blood flow, diffusely distributed neointimal plaques, appearing as discrete bulging towards the vascular lumen and medial thickening. Our findings suggest that increased CWT caused by hypertension play a pivotal role in the remodeling of the prestenotic segment through biomechanical effects on oxidative stress and increased expression of transforming growth factor beta. The second is remodeling in the normotensive poststenotic segment with turbulent flow/low wall shear stress and normal CWT and tensile stress characterized by groups of endothelial cells with phenotypic alterations and focally distributed neointimal plaques, similar but many of them larger than those found in the prestenotic segments. Further studies are needed to determine how the mechanical forces of turbulent flow/low shear stress are detected and transduced into chemical signaling by the cells of the artery walls and then converted into pathophysiologically relevant phenotypic changes. PMID- 16467655 TI - Effects of changes in sodium balance on plasma and kidney angiotensin II levels in anesthetized and conscious Ren-2 transgenic rats. AB - OBJECTIVE: Since there is as yet no general agreement regarding the role of plasma and kidney angiotensin II (ANG II) in the development of hypertension in Ren-2 transgenic rats (TGR), in the present study we evaluated plasma and kidney ANG II levels in anesthetized and conscious TGR and in normotensive Hannover Sprague-Dawley rats (HanSD) fed a normal salt diet (NS). Given the importance of ANG II in the development of salt-sensitive hypertension, and the fact that hypertensinogenic actions of ANG II are mediated via ANG II type 1 (AT1) receptors, the effects of high salt (HS) intake and of sodium depletion on blood pressure (BP), ANG II levels and kidney AT1 receptor protein expression in TGR and HanSD were also examined. METHODS: Rats were maintained on a NS diet (0.6% NaCl) or fed a HS diet (2% NaCl) for 4 days or were sodium depleted (40 mg/l furosemide for 1 day followed by 3 days of 0.01% NaCl diet). They were sacrificed either by an overdose of anesthetic (thiopental sodium) or by decapitation (without anesthetic) and plasma and kidney ANG II levels were determined by radioimmunoassay during the prehypertensive (32 days old), the early (52 days) and the maintenance (90 days) phases of hypertension. Total kidney AT1 receptor protein levels were assessed by Western blot analysis. RESULTS: In anesthetized animals fed the NS diet, plasma ANG II levels were lower in 32-day-old TGR than in HanSD, but at 52 and 90 days of age no significant differences were noted. ANG II concentrations in kidney tissue were similar in 32- and 90-day-old TGR and HanSD, but were higher in 52-day-old TGR than in HanSD. In contrast, in conscious animals immediately after decapitation, plasma and kidney ANG II levels were higher in TGR than in HanSD at all ages. HS diet did not change BP but suppressed ANG II levels in HanSD at all ages. In contrast, HS diet increased BP but did not decrease plasma and kidney ANG II levels in TGR at all ages. Sodium restriction did not alter BP and resulted in a marked increase in ANG II levels in HanSD, but caused a significant decrease in BP in TGR without altering plasma or tissue ANG II concentrations. There were no significant differences in renal AT1 receptor protein expression between HanSD and TGR at any age of any of the experimental groups. CONCLUSIONS: On the basis of our present results we conclude that TGR exhibit a disrupted interaction between sodium homeostasis and the regulation of the renin-angiotensin system (RAS) activity which results in the loss of BP regulation in this model. PMID- 16467656 TI - Conformational changes in prorenin during renin inhibition in vitro and in vivo. AB - BACKGROUND: Some renin inhibitors induce changes in the conformation of prorenin in vitro and influence the quantification of active renin by immunoradiometric assays. Whether such changes in renin recognition by monoclonal antibodies exist after oral administration of aliskiren, the first orally available renin inhibitor, is not known. METHODS: Two commercially available immunoradiometric assays (Cisbio and Nichols) were compared to determine immunoreactive active renin concentrations in plasma samples collected in a single oral dose crossover study comparing the renin inhibitor, aliskiren (300 mg), with the angiotensin II antagonist, valsartan (160 mg), in healthy male subjects. RESULTS: The addition of aliskiren to plasma samples in vitro, at concentrations of 1-100 micromol/l, increased active renin immunoreactivity in both the Cisbio and Nichols assays. In the crossover study, the two assays gave similar values for the plasma immunoreactive active renin concentration before treatment and following valsartan administration (intraclass coefficient for agreement between the two assays = 0.92). However, a Bland-Altman plot showed a systematic bias towards higher values (1.75-fold higher; 95% confidence interval = 1.02-3.01) in the Nichols than in the Cisbio assay following aliskiren administration. The difference between the results obtained with the two assays depended on incubation time. CONCLUSIONS: Depending on incubation conditions, circulating renin inhibitors interfere with the recognition of active renin molecules by the monoclonal antibodies used in commercially available assays. Careful consideration must therefore be given to the methodology used for quantifying immunoreactive plasma active renin when patients are treated with renin inhibitors, to avoid an overestimation of the magnitude of active renin release attributable to conformational changes in plasma prorenin. PMID- 16467658 TI - Microalbuminuria and tubular proteinuria as risk predictors of cardiovascular morbidity and mortality in essential hypertension: final results of a prospective long-term study (MARPLE Study)*. AB - OBJECTIVE: To evaluate the impact of microalbuminuria (MAU) or tubular proteinuria (TPU) on cardiovascular and cerebrovascular events and all-cause mortality, and to assess whether a normalization of MAU and/or TPU induced by angiotensin-converting enzyme-inhibitor-based antihypertensive treatment with ramipril improves cerebrovascular prognosis in essential hypertensive patients without diabetes mellitus. METHOD: A prospective, controlled, multicenter study was performed involving 3529 hypertensive participants (average follow-up 42.5 months). Ramipril was the basic antihypertensive medication. Proteinuria analysis (albumin, alpha 1-microglobulin, SDS electrophoresis) was performed by quantitative measurement every year. Ambulatory blood pressure monitoring was performed once yearly. The main outcome determined was cardiovascular and cerebrovascular events and all-cause mortality. RESULTS: In patients with TPU and/or MAU, the risk for endpoints increased significantly compared with normal (TPU, 30.0%; MAU, 54.7%; MAU + TPU, 64.0%; macroproteinuria, 74.4%). A change of protein excretion either from pathologic to normal or from normal to pathologic showed a clear trend to correlate with cerebrovascular endpoints (P = 0.056 and P = 0.055). Normal protein excretion at baseline and during follow-up indicated a significantly better prognosis than pathologic proteinuria at baseline and during follow-up. (P < 0.0001). TPU normalized in 31.9%, MAU in 30.6%, MAU + TPU in 29.3%, and macroproteinuria in 10.2% of patients. A total of 445 (25.4%) patients with normal protein excretion developed pathologic proteinuria during follow-up. CONCLUSIONS: In non-diabetic hypertensive patients, MAU as well as TPU increases the incidence of cardiovascular events. Normalization of MAU, TPU or macroproteinuria during angiotensin-converting enzyme-inhibitor-based treatment correlates with a reduction of cardiovascular events. Beyond blood pressure control, normalization of MAU and TPU should be considered as a further therapeutic goal. There is a need for further studies to optimize treatment if proteinuria is unresponsive to angiotensin-converting enzyme inhibitors. PMID- 16467657 TI - Hypertension after experimental cerebral ischemia: candesartan provides neurovascular protection. AB - BACKGROUND: After ischemic stroke, hypertension increases the risk of recurrence, hemorrhage and fatal cerebral edema, but blood pressure (BP) lowering in the acute stroke period is controversial due to fears of infarct extension and worsened outcomes. OBJECTIVE: To determine whether BP lowering with candesartan, initiated at reperfusion, can reduce neurovascular damage and improve outcome in a model of hypertension after experimental ischemic stroke. METHODS: Male Wistar rats (280-305 g) underwent 3 h of middle cerebral artery occlusion (MCAO). At reperfusion, either saline (n = 18) or candesartan 1 mg/kg (n = 18) was administered intravenously. BP was measured by telemetry for 2 days before and 24 h after MCAO. Neurologic function was assessed and sacrifice occurred at 24 h after occlusion. Brain tissue was analyzed for infarct size, hemoglobin content and edema. RESULTS: Mean BP increased from 96 to 124 mmHg immediately upon MCAO and decreased to 114 mmHg after reperfusion, remaining elevated for 24 h (P < 0.001) in the saline group. Candesartan reduced BP back to baseline and BP remained lower than in saline-treated animals until sacrifice (P < 0.001). Infarct size (54 versus 38%, P = 0.01) and hemoglobin content (23.4 versus 10.0 microg/g tissue; P = 0.03) and edema (17.97 versus 11.33%, P < 0.0001) were lower in the candesartan group. In addition, neurologic function at 24 h was improved (P = 0.0036) in the candesartan group. CONCLUSIONS: Candesartan administered after reperfusion in acute ischemic stroke reduces neurovascular damage and improves outcome. PMID- 16467659 TI - High-fat diet-induced juvenile obesity leads to cardiomyocyte dysfunction and upregulation of Foxo3a transcription factor independent of lipotoxicity and apoptosis. AB - BACKGROUND: Obesity is associated with dyslipidemia, which leads to elevated triglyceride and ceramide levels, apoptosis and compromised cardiac function. METHODS: To determine the role of high-fat diet-induced obesity on cardiomyocyte function, weanling male Sprague-Dawley rats were fed diets incorporating 10% of kcal or 45% of kcal from fat. Mechanical function of ventricular myocytes was evaluated including peak shortening (PS), time-to-PS (TPS), time-to-90% relengthening (TR90) and maximal velocity of shortening and relengthening (+/- dl/dt). Intracellular Ca properties were assessed using fluorescent microscopy. RESULTS: High-fat diet induced hyperinsulinemic insulin-resistant obesity with depressed PS, +/- dl/dt, prolonged TPS/TR90 reduced intracellular Ca release and Ca clearing rate in the absence of hypertension, diabetes, lipotoxicity and apoptosis. Myocyte responsiveness to increased stimulus frequency and extracellular Ca was compromised. SERCA2a and phospholamban levels were increased, whereas phosphorylated phospholamban and potassium channel (Kv1,2) were reduced in high-fat diet group. High-fat diet upregulated the forkhead transcription factor Foxo3a, and suppressed mitochondrial aconitase activity without affecting expression of the caloric sensitive gene silent information regulator 2 (Sir2), protein nitrotyrosine formation, lipid peroxidation and apoptosis. Levels of endothelial nitric oxide synthase (NOS), inducible NOS, triglycerides and ceramide were similar between the two groups. CONCLUSIONS: Collectively, our data show that high-fat diet-induced obesity resulted in impaired cardiomyocyte function, upregulated Foxo3a transcription factor and mitochondrial damage without overt lipotoxicity or apoptosis. PMID- 16467661 TI - Prevalence and correlates of aortic root dilatation in patients with essential hypertension: relationship with cardiac and extracardiac target organ damage. AB - OBJECTIVE: To assess the prevalence of aortic root dilatation in a large cohort of uncomplicated hypertensive patients and to evaluate the relations of aortic root size to different markers of cardiac and extracardiac target organ damage (TOD). METHODS: A total of 3366 untreated and treated essential hypertensive patients (mean age, 53 +/- 12 years) consecutively attending our out-patient hypertension clinic and included in the Evaluation of Target Organ Damage in Hypertension (an observational ongoing registry of hypertension-related TOD) were considered for this analysis. All patients underwent routine examinations, 24-h urine collection for microalbuminuria, echocardiography and carotid ultrasonography. RESULTS: Aortic root dilatation, defined by the sex-specific echocardiographic criteria of 40 mm in men and 38 mm in women, was present in 8.5% of men and in 3.1% of women. Compared with 3160 patients with normal aortic size, the group of 206 patients with an enlarged aortic root was older, had higher diastolic blood pressure values and included a greater fraction of subjects under antihypertensive treatment, with type 2 diabetes and metabolic syndrome. The prevalence of left ventricular hypertrophy, carotid intima-media thickening, plaques and microalbuminuria was significantly higher in patients with aortic root dilatation. According to a logistic regression analysis, left ventricular hypertrophy, carotid atherosclerosis, overweight and metabolic syndrome were the main independent and potentially modifiable predictors of aortic root dilatation in the whole hypertensive population as well as in untreated and treated hypertensive patients separately. CONCLUSIONS: Our study shows that hypertensive patients with aortic root enlargement have more pronounced alterations in cardiac structure and geometry as well as in carotid artery morphology compared with those without the enlargement. Aortic root dilatation therefore appears to be a useful marker of high cardiovascular risk related to TOD. Whether this alteration independently predicts cardiovascular morbidity remains to be proven. PMID- 16467660 TI - Pressure overload causes cardiac hypertrophy in beta1-adrenergic and beta2 adrenergic receptor double knockout mice. AB - OBJECTIVE: Cardiac hypertrophy arises as an adaptive response to increased afterload. Studies in knockout mice have shown that catecholamines, but not alpha1-adrenergic receptors, are necessary for such an adaptation to occur. However, whether beta-adrenergic receptors are critical for the development of cardiac hypertrophy in response to pressure overload is not known at this time. METHODS AND RESULTS: Pressure overload was induced by transverse aortic banding in beta1-adrenergic and beta2-adrenergic receptor double knockout (DbetaKO) mice, in which the predominant cardiac beta-adrenergic receptor subtypes are lacking. Chronic pressure overload for 4 weeks induced cardiac hypertrophy in both DbetaKO and wild-type mice. There were no significant differences between banded mice in left ventricular weight to body weight ratio, in the left ventricular wall thickness, in the cardiomyocyte size or in the expression levels of the load sensitive cardiac genes such as ANF and beta-MHC. Additionally, the left ventricular systolic pressure, an index of afterload, and cardiac contractility, evaluated as dp/dtmax, the maximal slope of systolic pressure increment, and Ees, end-systolic elastance, were increased at a similar level in both wild-type and DbetaKO banded mice, and were significantly greater than in sham controls. CONCLUSION: Despite chronic activation of the cardiac beta-adrenergic system being sufficient to induce a pathological hypertrophy, we show that beta1 adrenergic and beta2-adrenergic receptors are not an obligatory component of the signaling pathway that links the increased afterload to the development of cardiac hypertrophy. PMID- 16467662 TI - Treatment with beta-blockers, ACE inhibitors, and calcium-channel blockers is associated with a reduced fracture risk: a nationwide case-control study. AB - BACKGROUND: Cardiovascular diseases are associated with disturbances in calcium metabolism, including increased urinary calcium, vitamin D insufficiency, and decreased bone mineral density. Antihypertensive drugs may increase the risk of falling. However, risk of fracture in patients treated with non-diuretic cardiovascular drugs is largely unknown. AIM: We investigated associations between fracture risk and treatment with commonly used cardiovascular drugs: beta blockers, angiotensin-converting enzyme (ACE) inhibitors, and calcium-channel blockers. DESIGN: A population-based pharmaco-epidemiological case-control study with fracture in year 2000 as outcome and drug use during the previous 5 years as exposure. We used nationwide computerized registers to assess individual use of drugs and related these data to individual fracture records and information on socio-economic and health-related confounders. RESULTS: We included 124,655 cases that sustained a fracture and 373,962 age and gender-matched controls. After adjustment for potential confounders, risk of any fracture was reduced by 9% [odds ratio (OR) 0.91; 95% confidence interval (CI), 0.88-0.93] in users of beta blockers, by 6% (OR, 0.94; 95%CI, 0.91-0.96) in users of calcium-channel blockers, and by 7% (OR, 0.93; 95%CI, 0.90-0.96) in users of ACE inhibitors. Moreover, risk of hip fractures was reduced significantly by 7-14% in users of the three groups of drugs. No major differences were found between men and women or in subjects younger or older than 70 years of age. Sub-analyses indicated differences between groups of calcium-channel blockers, as use of non dihydropyridine drugs was associated with a larger risk reduction than use of dihydropyridine drugs. CONCLUSION: Treatment with beta-blockers, ACE inhibitors, and calcium-channel blockers is associated with a small but significantly reduced risk of fracture. PMID- 16467663 TI - Comparative effects of nebivolol and metoprolol on oxidative stress, insulin resistance, plasma adiponectin and soluble P-selectin levels in hypertensive patients. AB - OBJECTIVES: To determine the effects of nebivolol on oxidative stress, insulin resistance, adiponectin and plasma soluble P-selectin levels in hypertensive patients in comparison with metoprolol. MATERIAL AND METHODS: Eighty newly diagnosed hypertensive patients in grade 1 hypertension according to the European Society of Hypertension and European Society of Cardiology guidelines were enrolled in this prospective, blinded, randomized study. Seventy-two patients completed the study. After baseline assessment, each patient was randomly allocated to a 5 mg daily dose of nebivolol (n = 37, 20 male) or a 100 mg daily dose of metoprolol (n = 35, 18 male) and treated for 6 months. Blood pressure, heart rate, oxidative stress (malonyldialdehyde), homeostasis model assessment: insulin resistance, adiponectin and plasma soluble P-selectin levels were measured before and after treatment. RESULTS: At the end of treatment, nebivolol and metoprolol significantly decreased blood pressure and heart rate, with a more pronounced bradycardic effect of metoprolol. Nebivolol, but not metoprolol, significantly lowered oxidative stress (P = 0.03), the insulin resistance index (P = 0.003) and plasma soluble P-selectin levels (P = 0.008), and increased adiponectin levels (P = 0.04). CONCLUSION: Nebivolol, in contrast to metoprolol, improved oxidative stress, insulin sensitivity, decreased plasma soluble P selectin and increased adiponectin levels in hypertensive patients. These beneficial effects of nebivolol may contribute to a reduction in cardiovascular risk in hypertensive patients. PMID- 16467664 TI - Management of severe hypertension in a child with tuberous sclerosis-related major vascular abnormalities. AB - OBJECTIVE: We report the case of a 4-year-old girl with tuberous sclerosis who presented with severe renovascular hypertension secondary to compression of the renal arteries by a large infradiaphragmatic abdominal aortic aneurysm. DESIGN: Case report of a previously unreported case. RESULTS: Normotension could not be achieved despite conservative treatment with three antihypertensive drugs at high doses. An aortic graft was performed successfully but failed to correct the hypertension. Postoperatively, the child experienced acute renal failure after attempting treatment with additional enalapril. Further investigations revealed ischaemia of the left kidney and stenosis of the right renal artery. Subsequently, a combination of maximum dose minoxidil, a calcium antagonist, clonidine and peripheral alpha and beta-blockers, and diuretics was used, but her blood pressure control remained poor. A bypass between the hepatic artery and the right renal artery restenosed at the proximal anastomosis. Her renovascular hypertension was finally improved with an autotransplantation of her right kidney into the right iliac fossa. Fibromuscular dysplasia and abundant collagen deposition were the predominant features seen on histology. She is currently normotensive on moderate doses of a beta-blocker, amlodipine and a diuretic. CONCLUSION: Hypertension in this case was impossible to control medically, and the surgical approach was compromised by numerous complications. This case illustrates the challenges in the management of renovascular hypertension in a child with disease-associated vascular malformations. PMID- 16467665 TI - Brain pathology in a syndrome of hypertension, convulsion, and cerebral haemorrhage in thalassaemic patients after multiple blood-transfusions: a summary in reported Thai autopsy cases. PMID- 16467666 TI - Imaging the retina by en face optical coherence tomography. AB - PURPOSE: To present the possibilities of a new system that combines optical coherence tomography (OCT) and confocal ophthalmoscopy, producing en face OCT images in patients with retinal diseases. METHODS: A prototype OCT Ophthalmoscope (OTI, Toronto, Canada) was used to scan patients with retinal conditions. The system uses a super luminescent diode (lambda = 820 nm; Deltalambda = 20 nm) and currently scans at a rate of 2 frames per second. In each frame, the OCT Ophthalmoscope simultaneously produces a transversal OCT scan and a confocal image in the X/Y plane. Both images correspond pixel to pixel. RESULTS: Between January 2002 and August 2003, >800 patients with various retinal diseases were scanned with the OCT Ophthalmoscope. Illustrative cases with regularly seen macular diseases are presented, such as macular hole and central serous retinopathy. CONCLUSION: Current difficulties as well as future possibilities of this new en face OCT ophthalmoscope are discussed. By presenting normal and pathologic transversal OCT images made by a prototype OCT Ophthalmoscope, we show that it can provide information not available using conventional OCT imaging. PMID- 16467667 TI - Panretinal photocoagulation and intravitreal triamcinolone acetonide for the management of proliferative diabetic retinopathy with macular edema. AB - PURPOSE: To investigate intravitreal triamcinolone acetonide (IVTA) as an adjunctive therapy to panretinal photocoagulation (PRP) in patients with both high-risk proliferative diabetic retinopathy (PDR) and clinically significant macular edema (CSME). METHODS: Thirty-five eyes diagnosed with both high-risk PDR and CSME underwent PRP and a single injection of 4 mg of IVTA (IVTA group). Visual, anatomic, and fluorescein angiographic changes were documented. Any complications resulting from the combined procedure were noted. These data were compared retrospectively to 35 eyes that underwent grid laser treatment to the macula followed 2 weeks later by PRP (laser group). Main outcome measures included change in best-corrected visual acuity, improvement in macular edema (clinical or angiographic), and control of the neovascular disease. RESULTS: Mean follow-up was 9.6 months for the IVTA group and 11.9 months for the laser group. Mean pretreatment best-corrected visual acuity was 20/286 in the IVTA group and 20/282 in the laser group (P = 0.80). After 9 months of follow-up, visual acuity was 20/80 in the IVTA group versus 20/156 in the laser group (P = 0.007). Thirty four percent of eyes in the IVTA group had final vision of 20/40 or better versus 11% in the laser group (P = 0.044). At 9 months follow-up, 84% of IVTA eyes had complete resolution of macular edema versus 46% of laser eyes (P = 0.002). Three eyes in the IVTA group had recurrence of macular edema after 6 months and required reinjection of IVTA. Elevation in intraocular pressure occurred in eight eyes in the IVTA group and responded to topical therapy. Cataract progression was observed in nine eyes in the IVTA group. CONCLUSIONS: The addition of intravitreal triamcinolone acetonide to PRP in the management of patients with both PDR and CSME seems promising. Further study is needed to assess the effect of this combined treatment. PMID- 16467668 TI - Evaluating adjunctive surgical procedures during vitrectomy for diabetic macular edema. AB - PURPOSE: To evaluate the long-term outcomes of adjunctive surgical procedures during pars plana vitrectomy (PPV) for the treatment of diabetic macular edema (DME). METHODS: In this nonrandomized study, we retrospectively analyzed 57 eyes of 54 patients who had DME and had undergone PPV. We performed PPV using three different surgical procedures: conventional PPV (group PVD; 13 eyes), triamcinolone acetonide (TA)-assisted PPV (group TA; 22 eyes), and TA-assisted PPV combined with internal limiting membrane (ILM) peeling (group ILM; 22 eyes). We also evaluated the preoperative and postoperative best-corrected visual acuity (BCVA) results. RESULTS: The overall mean preoperative BCVA was 0.86 logarithm of the minimal angle of resolution unit. In groups PVD, TA, and ILM, BCVAs were 0.99, 0.90, and 0.74 (P = 0.310), respectively. The mean postoperative BCVA for all patients improved to 0.68 (P = 0.005). The postoperative BCVA improved in 47% of the treated eyes, it remained unchanged in 37% of the treated eyes, and it deteriorated in 16% of the treated eyes. However, we observed no significant difference in the mean postoperative BCVAs between the three groups. Furthermore, we found that there was no significant difference in postoperative BCVA improvements between any of the groups (P = 0.450). CONCLUSION: The present study suggests that these 3 PPV approaches do not significantly affect postoperative BCVAs after 18 months of DME treatment. PMID- 16467669 TI - Incidence of rhegmatogenous retinal detachment after vitrectomy in eyes of diabetic patients. AB - PURPOSE: To assess the incidence of rhegmatogenous retinal detachment (RRD) after pars plana vitrectomy (PPV) among diabetic patients with complications of proliferative diabetic retinopathy. METHODS: Ninety-three eyes of diabetic patients-who underwent PPV with or without intraocular gas tamponade for complications of proliferative diabetic retinopathy-were reviewed retrospectively. Indication for vitrectomy was vitreous hemorrhage in 80 patients (86.1%), tractional retinal detachment in 3 (3.2%), and vitreous hemorrhage associated with tractional retinal detachment in 10 (10.7%). RESULTS: Four (4.3%) of 93 eyes developed an RRD after vitrectomy. The primary reason for vitrectomy was recurrent or nonresolving vitreous hemorrhage. The retina was attached with one additional surgical procedure in two of these eyes; the other two had to undergo a third operation before attachment was achieved. CONCLUSION: RRD occurs in a small percentage of patients after PPV with or without gas tamponade for vitreous hemorrhage or tractional retinal detachment caused by proliferative diabetic retinopathy. Thorough postoperative follow-up is important to make early diagnosis and intervention possible. PMID- 16467670 TI - Stereoscopic fluorescein angiography in diabetic maculopathy. AB - PURPOSE: To explore the possible differences in the depth location of microaneurysms in focal as well as diffuse diabetic macular edema. METHODS: The density of superficial and deep retinal microaneurysms was assessed using a stereoscopic fluorescein angiographic method. RESULTS: The density of deep retinal microaneurysms was nearly identical in the group with diffuse macular edema (89.9 +/- 28.5 microaneurysms/test grid) and the group with focal macular edema (90 +/- 65.4 microaneurysms/test grid). The density of superficial retinal microaneurysms was significantly lower in the group with diffuse macular edema (22.8 +/- 12.5) than in the group with focal macular edema (47.9 +/- 30.6). The density fraction of superficial retinal microaneurysms was lower in the group with diffuse macular edema (0.19 +/- 0.07) than in the group with focal macular edema (0.36 +/- 0.14). In the group with focal macular edema, areas without edema had a significantly lower density of superficial (6.1 +/- 2.8) and deep (15.6 +/- 7.8) retinal microaneurysms than did areas with edema (superficial: 47.9 +/- 30.6; deep: 90.0 +/- 65.4). In the group with focal macular edema, the density fraction of superficial retinal microaneurysms was nearly identical in areas with (0.36 +/- 0.14) and areas without (0.30 +/- 0.10) edema. CONCLUSION: The development of focal macular edema might be linked to the density of microaneurysms. Diffuse macular edema might be a result of some unknown effect on the deeper retina and/or the choroid. PMID- 16467671 TI - Indocyanine green angiographic findings for patients with systemic lupus erythematosus nephropathy. AB - PURPOSE: To study indocyanine green (ICG) angiographic findings for patients with systemic lupus erythematosus (SLE) nephropathy. In particular, the presence of choroidal abnormalities at ICG angiography, which could not be detected by fluorescein angiography, was studied. DESIGN: Observational case report study. METHODS: Nine consecutive female patients (mean age +/- SD, 38.2 +/- 9.7 years) with SLE-related nephropathy underwent simultaneous ICG angiography and fluorescein angiography according to a standard angiographic protocol for uveitis. RESULTS: Two findings were revealed by ICG angiography: focal, transient hypofluorescent areas in the early phase; and spots of choroidal hyperfluorescence visible from the intermediate to late phase. CONCLUSIONS: ICG angiography can provide information that is not detectable by clinical or fluorescein angiographic examination for patients with lupus nephropathy. This information may prove useful in better understanding the pathogenesis of SLE choroidopathy. PMID- 16467672 TI - Intraocular viral and immune pathogenesis of immune recovery uveitis in patients with healed cytomegalovirus retinitis. AB - PURPOSE: To investigate immune and viral contributions to the pathogenesis of immune recovery uveitis (IRU), which presents as vitritis, macular edema, or formation of epiretinal membranes, and develops in patients with acquired immunodeficiency syndrome (AIDS) who experienced cytomegalovirus (CMV) retinitis before antiretroviral treatment (ART) induced immune reconstitution. METHODS: Aqueous and vitreous fluids from patients with IRU, active CMV retinitis, and control human immunodeficiency virus (HIV)-negative, noninflamed eyes were compared for presence of cytokines IL-6, IL12, interferon gamma using enzyme linked immunosorbent assay techniques, and CMV DNA (by polymerase chain reaction). RESULTS: IRU eyes (11 patients, 18 samples) had the highest levels of IL-12 (median 48 pg/mL), moderate levels of IL-6 (median 146 pg/mL), and low but significant interferon gamma (median 15 pg/mL), compared to controls (P < 0.01). All uveitis eyes tested (9/9) were CMV DNA negative. In contrast, active CMV retinitis eyes were CMV DNA positive, had higher levels of IL-6 (median 349 pg/mL) (25 patients, 41 samples) than both control (P = 0.0001) and uveitis eyes (P = 0.048), similar levels of interferon gamma (median 27 pg/mL) to uveitis eyes, but less IL-12 (median 0 pg/mL) than uveitis eyes. CONCLUSIONS: Inflammatory IRU can be differentiated from active CMV retinitis by the presence of IL-12, less IL-6, and absence of detectable CMV replication. PMID- 16467673 TI - High prevalence of myopia in Japanese patients with idiopathic focal subretinal neovascularization. AB - PURPOSE: To determine whether myopia is more prevalent in Japanese patients with idiopathic focal subretinal neovascularization (IFSN) than in normal control subjects. METHODS: Forty-seven eyes of 46 patients with an initial diagnosis of IFSN and 291 eyes of 291 controls were studied. Refractive errors were measured with an autorefractometer, and the spherical equivalent of the refractive error was used for the statistical analyses. All patients had undergone fluorescein angiography (FA) to confirm the presence of the choroidal neovascularization. In addition, indocyanine green angiography (ICGA) had been performed to determine whether chorioretinal atrophy and breaks of Bruch's membrane, which are consistent with myopic retinopathy, were present. RESULTS: The mean age of the patients in the control group was not significantly different from that of patients in the IFSN group. The mean spherical equivalent of the refractive errors was -2.62 +/- 2.70 diopters (D) in the control group and -5.24 +/- 3.41 D in the IFSN group (P = 0.00005). The incidence of high myopia was significantly higher in the IFSN group (41.3%) than in the control group (12.0%, P < 0.0001). FA and ICGA showed no chorioretinal atrophy and lacquer cracks, thus confirming that patients with myopic retinopathy were excluded from the IFSN group. During the mean follow-up period (44.7 months), myopic retinopathy did not develop in any of the eyes in the IFSN group. CONCLUSION: Japanese patients with IFSN were highly myopic, suggesting that myopia may play a role in the development of choroidal neovascularization in IFSN patients. PMID- 16467674 TI - Posterior pole retinal thickness measurements by the retinal thickness analyzer in healthy Chinese subjects. AB - PURPOSE: To assess retinal thickness at the posterior pole in healthy Chinese subjects with the retinal thickness analyzer (RTA). METHODS: Retinal thicknesses at the posterior pole and fovea were measured by the RTA in 331 eyes of 178 healthy Chinese subjects. Retinal thicknesses as a function of sex, age, refractive errors, and axial length were also evaluated. RESULTS: The average thickness of the foveal area +/- SD was 158.6 +/- 24.8 microm, the average thickness of the perifoveal region (600-2,500 microm from fovea) +/- SD was 174 +/- 25.2 microm, and the average thickness of the posterior pole region (600 6,000 microm from fovea) +/- SD was 171.9 +/- 25.3 microm. There was no significant difference in retinal thicknesses of the foveal, perifoveal, and posterior pole regions in either right or left eyes or as a function of different spherical equivalents. Moreover, there was no significant difference of foveal thickness between males and females. Greater retinal thicknesses of the foveal, perifoveal, and posterior pole regions were associated with age of older than 40 years and axial length of < 24 mm, and greater perifoveal and posterior pole thicknesses were found for females. Furthermore, there were no significant correlations between age, refractive errors, and axial lengths with retinal thicknesses. CONCLUSION: Retinal thicknesses of the posterior pole region differed by age, axial length, and sex, but there was no effect of laterality or different spherical equivalents. The RTA is a fast and noninvasive technology to assess retinal conditions with thickening or thinning in the fovea and posterior pole. PMID- 16467675 TI - Intravitreal toxicity of garenoxacin. AB - PURPOSE: To assess the retinal toxicity of varying concentrations of intravitreally injected garenoxacin. METHODS: Twenty eyes of 20 New Zealand albino rabbits were used for this study. The animals were anesthetized with ketamine (35-50 mg/kg) and xylazine (3-5 mg/kg). Garenoxacin was titrated using distilled water to the following concentrations: 4,000, 2,000, 1,000, 400, 200, and 100 microg/0.1 mL. Each concentration was injected intravitreally (0.1 mL) into three rabbit eyes. Three control eyes were injected with 0.1 mL of balanced saline solution. All animals were examined before and after injection by indirect ophthalmoscopy and slit-lamp biomicroscopy. Electroretinography was performed on all animals before intravitreal injection and 14 days after injection. The animals were examined by indirect ophthalmoscopy and slit-lamp biomicroscopy before they were killed; the eyes were enucleated and examined with light microscopy. RESULTS: No electroretinographic changes or signs of retinal toxicity by slit-lamp examination, indirect ophthalmoscopy, or light microscopy were seen in any eyes 14 days after intravitreal injection of garenoxacin (< or =4,000 microg/0.1 mL). CONCLUSIONS: Garenoxacin injected intravitreally appeared safe at concentrations of < or =4,000 microg/0.1 mL. PMID- 16467676 TI - Intravitreal toxicity of moxifloxacin. AB - PURPOSE: To assess the retinal toxicity of various concentrations of intravitreally administered moxifloxacin, a fourth-generation fluoroquinolone. METHODS: Ten New Zealand albino rabbits were divided into five groups. The initial concentration of moxifloxacin (400 mg/250 mL) was titrated using 5% dextrose solution to concentrations (320 microg/0.1 mL, 160 microg/0.1 mL, 100 microg/0.1 mL, and 50 microg/0.1 mL) that were injected intravitreally into 1 eye of each rabbit. Two control eyes were injected intravitreally with 0.1 mL of 5% dextrose solution. All animals were examined before and after injection by indirect ophthalmoscopy and slit-lamp biomicroscopy; electroretinography (ERG) was performed on all animals. The animals were killed, and their eyes were enucleated and examined with light microscopy. RESULTS: Remarkable decreases in ERG findings were noted in the group injected with moxifloxacin at a concentration of 320 microg/0.1 mL. No meaningful ERG changes were observed in eyes injected with moxifloxacin at other concentrations. There were no signs of retinal toxicity during slit-lamp examination, indirect ophthalmoscopy, or light microscopy in any eyes injected with moxifloxacin concentrations of < or =160 microg/0.1 mL. CONCLUSIONS: Intravitreal injection of moxifloxacin at a concentration of < or =160 microg/0.1 mL appeared nontoxic in the rabbit eye. PMID- 16467677 TI - Vitreous penetration of topical moxifloxacin and gatifloxacin in humans. AB - PURPOSE: To determine the vitreous penetration of the new fourth-generation topical fluoroquinolones moxifloxacin 0.5% and gatifloxacin 0.3%. METHODS: A prospective randomized clinical trial comprising 12 eyes of 12 patients scheduled for pars plana vitrectomy between August 2003 and September 2003 was performed in a clinical practice. The patients were randomly assigned to receive topical moxifloxacin 0.5% (n = 6) or gatifloxacin 0.3% (n = 6). One half the patients in each antibiotic group received 1 drop every 15 minutes for a total of 3 doses starting 1 hour before surgery, and the other one half self-administered the antibiotic drop 4 times daily for 3 days before surgery and at 7 am on the day of surgery. Undiluted vitreous samples were obtained and analyzed using high performance liquid chromatography. RESULTS: Either moxifloxacin 0.5% or gatifloxacin 0.3% was detected in the vitreous in all 12 patients in the study. There was no significant difference between the mean vitreous concentration of moxifloxacin 0.5% given over 1 hour preoperatively (0.012 +/- 0.011 microg/mL) and that given in the 3-day regimen (0.011 +/- 0.008 microg/mL) (P = 0.93). There was also no significant difference between the mean vitreous concentration of gatifloxacin 0.3% given over 1 hour preoperatively (0.001 +/- 0.0003 microg/mL) and that given over 3 days (0.008 +/- 0.006 microg/mL) (P = 0.11). Vitreous concentrations of moxifloxacin 0.5% and gatifloxacin 0.3% in each eye were all lower than the 90% minimum inhibitory concentration for the commonest bacterial isolates causing endophthalmitis. With both dosing regimens, the mean vitreous concentration of moxifloxacin 0.5% was higher than that of gatifloxacin 0.3% administered at the same regimen, but this was not statistically significant. CONCLUSION: Both topical moxifloxacin 0.5% and gatifloxacin 0.3% penetrated the vitreous in the uninflamed eye, but the vitreous concentrations attained were all lower than the 90% minimum inhibitory concentration for the commonest bacterial pathogens causing acute postoperative endophthalmitis. PMID- 16467678 TI - Neuroprotective effects of intramuscular ketamine in rabbit retinas after pars plana vitrectomy and silicone oil injection. AB - PURPOSE: To investigate potential retinal neuroprotective effects of intramuscular ketamine in rabbits after pars plana vitrectomy (PPV) and intravitreal silicone oil injection (SOI). METHODS: Twelve New Zealand rabbits (weight, 2.0-2.5 kg) underwent PPV with SOI in the right eye. Postoperatively, six rabbits received a daily intramuscular injection of ketamine for 4 weeks (ketamine-operated eyes), and six rabbits received a daily intramuscular injection of saline (saline-operated eyes). The retina from the left eye of each rabbit served as a control (ketamine-control and saline-control eyes). The animals were euthanized at 4 weeks after surgery. Qualitative and quantitative analyses were performed using the Zeiss Axiophot microscope and KS 400 software. RESULTS: Qualitative analysis using light microscopy demonstrated more extensive edema and cell disorganization in saline-operated retinas than in ketamine operated, ketamine-control, and saline-control retinas. Quantitatively, the cell densities (cell/mm) in the outer nuclear layer (ONL), inner nuclear layer (INL), and ganglion cell layer (GCL) in saline-operated retinas were significantly (P < 0.05) lower than those in these layers in ketamine-operated, ketamine-control, and saline-control retinas. The cell density in the ONL in saline-operated retinas was 52% lower than that in ketamine-operated retinas, 55% lower than that in ketamine-control retinas, and 56% lower than that in saline-control retinas. The cell density in the INL in saline-operated retinas was 44% lower than that in ketamine-operated retinas, 48% lower than that in ketamine-control retinas, and 49% lower than that in saline-control retinas. The cell density in the GCL in saline-operated retinas was 60% lower than that in ketamine-operated retinas, 64% lower than that in ketamine-control retinas, and 64% lower than that in saline control retinas. CONCLUSION: PPV with SOI was associated with retinal cell death and disorganization in rabbit eyes. Intramuscular ketamine administration provided protection against these effects. PMID- 16467679 TI - Effect of a novel octapeptide urokinase fragment, A6, on experimental choroidal neovascularization in the monkey. AB - PURPOSE: To evaluate the inhibitory effects of a urokinase-derived octapeptide, A 6, on laser-induced choroidal neovascularization (CNV) in monkeys. METHODS: Twenty female cynomolgus monkeys were randomly grouped into weekly or monthly A 6 treatment groups, each consisting of 10 animals. CNV was induced in both eyes by perimacular laser treatment. In each right eye, a single 22.25-mg A 6 dose (monthly group) or 4 22.25-mg A 6 doses each week (weekly group) were given by intravitreal injections. Each left eye received phosphate buffer on the same schedule. Monkeys were observed for 4 weeks by ophthalmic examinations, color photography, and fluorescein angiography. RESULTS: Weekly treated eyes had a 35% reduction of CNV compared with controls (P = 0.23). In contrast, monthly treated eye had a 71% reduction of CNV compared with controls (P = 0.0009). There was no evidence of toxicity at both clinical and pathologic examinations. CONCLUSIONS: Intravitreal A 6 injections effectively inhibited CNV in cynomolgus monkeys without evidence of toxicity. The overall reduction in CNV was greater for monthly treated eyes than for weekly treated eyes. This study suggests that A 6 has promise as a local antiangiogenic treatment of CNV. Further work is indicated to evaluate the potential role of A 6 in therapy for human CNV associated with age-related macular degeneration. PMID- 16467680 TI - Prevention of experimental proliferative vitreoretinopathy with a biodegradable intravitreal drug delivery system of all-trans retinoic acid. AB - PURPOSE: To evaluate the antiproliferative effect of an all-trans retinoic acid (at-RA) drug delivery system (DDS) on experimental proliferative vitreoretinopathy (PVR). METHODS: PVR was induced in rabbits with core vitrectomy and fibroblast injection. The DDS containing 420 microg, 650 microg, and 1,070 microg of at-RA was implanted into the vitreous of treated groups B, C, and D, respectively. Group A with no DDS and group E with nonmedicated DDS served as controls. The intravitreal at-RA concentration was measured with high-pressure liquid chromatography. The drug toxicity was evaluated histologically. RESULTS: The severity of PVR was significantly reduced in groups C and D but not in groups A, B, and E. The drug release peaked at 6 weeks to 7 weeks. No signs of retinal toxicity were found in the DDS groups. CONCLUSION: Intravitreal implantation of at-RA DDS appears effective in inhibiting the development of PVR and is well tolerated in rabbit eyes. PMID- 16467681 TI - Diagnostic and therapeutic challenges. PMID- 16467682 TI - Diagnostic and therapeutic challenges. PMID- 16467683 TI - Double use of optic fiber during vitrectomy as a transilluminator and indenter instrument. PMID- 16467684 TI - Chronic endophthalmitis due to verticillium species after cataract surgery treated (or managed) with pars plana vitrectomy and oral and intravitreal voriconazole. PMID- 16467685 TI - Intravitreal triamcinolone acetonide in Eales disease. PMID- 16467686 TI - Nontraumatic macular holes associated with Terson syndrome. PMID- 16467687 TI - Contact lens suturing for prevention of flap displacement during scleral buckling for retinal detachment in patients who have undergone laser in situ keratomileusis. PMID- 16467688 TI - Optical coherence tomography in the diagnosis of premacular hemorrhage. PMID- 16467689 TI - Intravitreal triamcinolone for ischemic branch retinal vein occlusion with serous retinal detachment. PMID- 16467690 TI - Use of perfluorocarbon liquid to prevent contact between indocyanine green and retinal pigment epithelium during surgery for idiopathic macular hole. PMID- 16467691 TI - Subretinal trypan blue migration during epiretinal membrane peeling. PMID- 16467692 TI - Choroidal neovascularization complicating photodynamic therapy for central serous retinopathy. PMID- 16467693 TI - Surgical repair of a retinal detachment in a patient with osteogenesis imperfecta. PMID- 16467694 TI - Rapid recovery of sympathetic ophthalmia with treatment augmented by intravitreal steroids. PMID- 16467695 TI - Autofluorescence and visual field loss in sector retinitis pigmentosa. PMID- 16467697 TI - Incidence of early and late sclerotomy-related retinal breaks in patients who underwent pars plana vitrectomy. PMID- 16467698 TI - Assessment of intraoperative safety in transoral robotic surgery. AB - INTRODUCTION: Robotic technology has been safely integrated into thoracic and abdominopelvic surgery, and the early experience has been very promising with very rare complications related to robotic device failure. Recently, several reports have documented the technical feasibility of transoral robotic surgery (TORS) with the daVinci Surgical System. Proposed pharyngeal and laryngeal applications include radical tonsillectomy, base-of-tongue resection, supraglottic laryngectomy, and phonomicrosurgery. The safety of transoral placement of the robotic endoscope and instruments has not been established. Potential risks specific to the transoral use of the surgical robot include facial skin laceration, tooth injury, mucosal laceration, mandible fracture, cervical spine fracture, and ocular injury. We hypothesize that these particular risks of transoral surgery are similar with robotic assistance compared with conventional transoral surgery. METHODS: To test this hypothesis, we attempted to intentionally injure a human cadaver with the daVinci Surgical System by impaling the facial skin and pharyngeal and laryngeal mucosa with the robotic instruments and endoscope. We also attempted to extract or fracture teeth and fracture the cadaver's mandible and cervical spine by applying maximal pressure and torque with the robotic arms. Experiments were documented with still and video photography. RESULTS: Impaling the cadaver's skin and mucosa resulted in only superficial lacerations. Tooth, mandible, and cervical spine fracture could not be achieved. CONCLUSIONS: Initial experiments performing TORS on a human cadaver with the daVinci Surgical System demonstrate a safety profile similar to conventional transoral surgery. Additionally, we discuss several strategies to increase patient safety in TORS. PMID- 16467699 TI - Primary treatment of ranula with intracystic injection of OK-432. AB - OBJECTIVE: Although surgery is the first choice of therapy for ranula, it was made a hypothesis that ranula can be primarily treated with sclerotherapy from prior evidence. This study examined the effectiveness of intracystic injection of OK-432 for treatment of ranula. METHOD: This prospective clinical study comprised a total of 26 patients with ranula (19 intraoral type; seven plunging type) treated with OK-432 sclerotherapy. Aspirated mucus of ranula was replaced with an equal volume of OK-432 solution of 0.01 mg/mL. The size of ranula was compared before and after sclerotherapy. RESULTS: Twenty of 26 patients (77%) showed a complete response after sclerotherapy: higher in plunging ranula (86%) than in intraoral ranula (74%). Rupture of ranula developed in seven of 19 patients (37%) with intraoral ranula within a few days after injection. The early rupture occurred more frequently in patients having a less-than-marked response and seemed to cause an increase in the total number of OK-432 injections: seven ruptured cases versus 12 nonruptured cases (mean 3.6 versus 1.5, P<.001). Recurrence occurred in two patients during a median follow-up period of 12 months (range, 9-22 mo) after the last injection. There were no major side effects, scarring, or increased morbidity to surgery of the OK-432-injected lesions. CONCLUSION: The intracystic injection of OK-432 is highly effective as a primary treatment modality of ranula. PMID- 16467700 TI - Morbidity after flap reconstruction of hypopharyngeal defects. AB - OBJECTIVES: Laryngopharyngeal reconstruction continues to challenge in terms of operative morbidity and optimal functional results. The primary aim of this study is to determine whether complications can be predicted on the basis of reconstruction in patients undergoing pharyngectomy for tumors involving the hypopharynx. In addition, we detail a reconstructive algorithm for management of partial and total laryngopharyngectomy defects. METHOD: A retrospective review was performed of 153 patients undergoing flap reconstruction for 85 partial and 68 circumferential pharyngectomies at a single institution over a 10-year period. There were 118 males and 35 females, the median age was 62 years, and mean follow up was 3.1 years. Pharyngectomy was performed for recurrence after radiotherapy in 80 patients and as primary surgery in 73. Free flap reconstruction was used in 42%, with 30 jejunal, 15 radial forearm, 11 anterolateral thigh, five rectus abdominis, and three gastro-omental flaps. Gastric transposition and pectoralis major pedicle flap was used in 14% and 44% of patients, respectively. Morbidity was analyzed according to extent of defect, regional versus free flap, enteric versus fasciocutaneous free flap reconstruction, and the effect of laparotomy. RESULTS: The total operative morbidity and mortality rate was 71% and 3%, respectively. The most common complications were hypocalcemia in 45%, pharyngocutaneous fistula in 33%, and wound complications in 25%. The late complication and stricture rate was 26% and 15%, respectively. On univariate analysis, circumferential defects were associated with increased total (P=.046) and flap-related morbidity (P=.037), hypocalcemia (P<.001), late complications (P=.003), and stricture (P=.009). Gastric transposition had increased total (P=.007), flap-related (P=.035), late complications (P=.034), and hypocalcemia (P=.001). Pharyngocutaneous fistula was increased in patients undergoing salvage pharyngectomy for radiation failure (P=.048) compared with primary surgery. On multivariate analysis, gastric transposition independently predicted for wound complications (P=.014) and fistula (P=.012). Circumferential defects predicted for flap-related morbidity (P=.030), hypocalcemia (P=.017), and late complications (P=.042). Tracheoesophageal speech was the method of voice restoration in 44% of patients. Oral diet was achieved in 93% of patients; however, 16% required gastrostomy tube feeds for either total or supplemental nutrition. CONCLUSION: The operative morbidity associated with pharyngeal reconstruction is substantial in terms of early and late complications. We were able to predict morbidity by defect extent and reconstruction type and initial treatment modality. Swallowing function is acceptable; however, less than half of the patients undergoing pharyngectomy had tracheoesophageal puncture voice restoration. PMID- 16467701 TI - The Edwin Smith Papyrus: the birth of analytical thinking in medicine and otolaryngology. AB - The Edwin Smith Papyrus, discovered in 1862 outside of Luxor, Egypt, is the oldest known surgical text in the history of civilization. The surviving scroll, a copy of an earlier text from around 3,000 B.C., gives us remarkable insight into the medical practice of ancient Egyptians in the Nile River bed during the dawn of civilization. The Papyrus is divided into 48 cases, most of which describe traumatic injuries. The text instructs the physician to examine the patient and look for revealing physical signs that may indicate the outcome of the injury. Although in modern medicine we take for granted that the use of physical examination and rational thinking lead to an accurate conclusion, 5,000 years ago, this was extraordinary. The Edwin Smith Papyrus cast aside the prevailing magic and mysticism of that time in favor of logic and deductive reasoning. As Egyptian civilization declined during the next millennium, the teachings of the Papyrus would be lost. It would not be until 300 B.C. when Hippocrates and his disciples in ancient Greece would once again revive logic in medical thinking and teaching. It is believed that the ancient Greeks had knowledge of the contents of the Edwin Smith Papyrus and its teachings and used them as the basis for their writings. As Europe entered the Dark Ages, so did medicine yet again, reverting to spells and prayers instead of judgment and reason. Although Hippocrates teachings were recognized by some scholars during the Middle Ages, they did not make up the basis for mainstream medical knowledge. With the dawn of the Renaissance, medicine would finally purge itself of its past supernatural foundation. Hippocratic teachings were used to form the basis of modern medicine, and medical pioneers in the 17 century studied the ancient Greek texts as the origin for their ideas. Many of the concepts physicians and patients today take as common knowledge originated in the Edwin Smith Papyrus. The authors attempt to uncover some of these fundamental ideas and trace them through time until their incorporation in our modern medical knowledge base. It is the rational, logical, and advanced thinking exhibited in the Edwin Smith Papyrus that mandates its respect from modern otolaryngologists and all physicians alike. PMID- 16467702 TI - A double-blind, randomized, placebo-controlled trial of macrolide in the treatment of chronic rhinosinusitis. AB - OBJECTIVES: The antiinflammatory effect of macrolide antibiotics has been well established, as has their role in the treatment of certain disorders of chronic airway inflammation. Several studies have suggested that long-term, low-dose macrolides may be efficacious in the treatment of chronic rhinosinusitis; however, these studies have lacked a control group. To date, this effect has not been tested in a randomized, placebo-controlled study. METHOD: The authors conducted a double-blind, randomized, placebo-controlled clinical trial on 64 patients with chronic rhinosinusitis. Subjects received either 150 mg roxithromycin daily for 3 months or placebo. Outcome measures included the Sinonasal Outcome Test-20 (SNOT-20), measurements of peak nasal inspiratory flow, saccharine transit time, olfactory function, nasal endoscopic scoring, and nasal lavage assays for interleukin-8, fucose, and a2-macroglobulin. RESULTS: There were statistically significant improvements in SNOT-20 score, nasal endoscopy, saccharine transit time, and IL-8 levels in lavage fluid (P<.05) in the macrolide group. A correlation was noted between improved outcome measures and low IgE levels. No significant improvements were noted for olfactory function, peak nasal inspiratory flow, or lavage levels for fucose and a2-macroglobulin. No improvement in any outcome was noted in the placebo-treated patients. CONCLUSION: These findings suggest that macrolides may have a beneficial role in the treatment of chronic rhinosinusitis, particularly in patients with low levels of IgE, and supports the in vitro evidence of their antiinflammatory activity. Additional studies are required to assess their place in clinical practice. PMID- 16467703 TI - Epiglottic and base-of-tongue prolapse in children: grading and management. AB - OBJECTIVES: A distinct entity of airway obstruction from epiglottic and base-of tongue (EBT) prolapse in the pediatric population is defined. Laryngopharyngeal findings, swallowing dysfunction, and gastroesophageal reflux disease are described in a group of children with EBT prolapse. A new grading system is also presented. STUDY DESIGN: A prospective study was conducted of laryngopharyngeal findings in children with EBT prolapse, a description of a new grading system, and review of the pediatric literature. METHODS: Fourteen children with EBT prolapse were prospectively studied with flexible fiberoptic nasopharyngolaryngoscopy. A new grading system for EBT prolapse was developed. Grade 0 is a normal airway. Grade 1 is prolapse of the epiglottis against the posterior pharyngeal wall but with normal position of the tongue. Grade 2 is prolapse of the epiglottis and base of tongue with only the epiglottic tip visible. Grade 3 is glossoptosis with no portion of the epiglottis visible. The diagnostic modalities and treatments for EBT prolapse are reviewed. RESULTS: Of the 14 children studied, seven (50%) had grade 1, four (29%) had grade 2, and three (21%) had grade 3 EBT prolapse. Swallowing dysfunction (five of 14, 38%) and gastroesophageal reflux disease (13 of 14, 93%) were also prevalent. CONCLUSION: The new grading system was applied successfully to describe the severity and sites of airway obstruction in 14 children with EBT prolapse. Swallowing dysfunction and gastroesophageal reflux disease occur in this population. Although surgical therapies are sometimes effective, lack of consistent success and the risk of aspiration with procedures other than tracheotomy may lead some to conclude that continuous positive airway pressure or tracheotomy are the safest options. PMID- 16467705 TI - Signaling pathways in interleukin-1beta-mediated middle ear mucin secretion. AB - OBJECTIVES: The objectives of this study were to investigate the role of the phosphatidylcholine-specific phospholipase C (PC-PLC), protein kinase C (PKC), and nitric oxide synthase (NOS) pathways during upregulation of mucin secretion by middle ear epithelium after exposure to interleukin-1beta and to examine the ability of a specific interleukin-1 receptor antagonist (IL-1betara) to block this increased secretion. MATERIALS AND METHODS: Primary chinchilla middle ear epithelial cultures were established and exposed to IL-1beta. Specific inhibitors of calmodulin, PC-PLC, PKC, and NOS pathways were used to investigate the potential role of these pathways leading to increased epithelial mucin secretion after exposure to IL-1beta. Mucin secretion was characterized by exclusion chromatography and liquid scintillation. RESULTS: Epithelial cultures exposed to IL-1beta demonstrate an increase in mucin secretion that is blocked by specific inhibitors of PC-PLC, PKC, and NOS, but not by inhibitors of calmodulin. In addition, mucin secretion stimulated by IL-1beta was reversible with use of a specific IL-1betara. CONCLUSIONS: IL-1beta stimulates mucin secretion from middle ear epithelium and its effects can be reversed by IL-1betara. PC-PLC, PKC, and NOS pathways play a role in the increased secretion of mucin in middle ear epithelial cells after exposure to IL-1beta. PMID- 16467704 TI - Osteoprotegrin knockout mice demonstrate abnormal remodeling of the otic capsule and progressive hearing loss. AB - OBJECTIVES: The otic capsule, when compared with other bones in the body, is unique in that it undergoes no significant remodeling of bone after development. We previously demonstrated that osteoprotegerin (OPG), which inhibits formation and function of osteoclasts, is produced at high levels in the inner ear of normal mice and secreted into the perilymph from where it diffuses into the surrounding otic capsule bone through a lacunocanalicular system. To test our hypothesis that the high level of OPG may be important in the inhibition of otic capsule remodeling, we studied the light microscopic histology of the otic capsule in OPG knockout mice for evidence of abnormal remodeling of bone. We also tested the hearing in OPG knockout mice to determine whether OPG and its influence on surrounding bone is important for auditory function. METHODS: Temporal bone histopathology and pathophysiology were compared in homozygous OPG knockout mice and C57BL/6 (B6) mice, the background strain for the knockouts. Auditory function in age-matched animals from each group was evaluated at approximately 4-week intervals from 8 to 21 weeks using frequency-specific auditory brainstem responses (ABR) and distortion product otoacoustic emissions (DPOAE). After each of the last three evaluations, the cochleae from one mouse of each group were harvested, processed, and examined by light microscopy. RESULTS: Osteoprotegerin knockout mice demonstrated abnormal remodeling of bone within the otic capsule with multiple foci showing osteoclastic bone resorption and formation of new bone. Such changes were not seen in the age-matched B6 controls. The active bone remodeling process in the knockout animals showed many similarities to otosclerosis seen in human temporal bones. Over the time period that we monitored, auditory function was significantly and progressively compromised in the knockout animals relative to B6 controls. At the earliest age of test (8 wk), the loss was apparent as a mild, high-frequency reduction in sensitivity by ABR. In contrast, DPOAE losses in the knockouts were substantial even at 8 weeks, and by 21 weeks, these losses exceeded our equipment limits. Results of ABR testing showed hearing sensitivity changes in the animals of the background strain were confined largely to the high frequencies, whereas OPG knockouts demonstrated substantial low-frequency shifts in addition to those at high frequencies. CONCLUSIONS: The histopathological and pathophysiological findings in OPG knockout mice support the hypothesis that OPG is important in the inhibition of bone remodeling within the otic capsule and the maintenance of normal auditory function. This mouse may provide a valuable animal model of human otosclerosis. PMID- 16467706 TI - Cranial bone regeneration using a composite scaffold of Beta-tricalcium phosphate, collagen, and autologous bone fragments. AB - OBJECTIVE: The aim of our study was to examine the tissue response and new bone formation induced by beta-tricalcium phosphate (beta-TCP), collagen, and autologous bone fragments with fibrin glue implanted into a cranial bone defect. MATERIALS AND METHODS: Granulated beta-TCP and collagen from porcine skin were mixed and freeze-dried. The weight ratio of both materials was 98:2. The composite scaffold for bone regeneration was composed of this artificial material and autologous bone fragments. Five adult beagle dogs were used. A cranial bone defect (2 cmx2 cm) was created in each dog. The animals were divided into two groups. In group 1 (n=3), the cranial bone defect was closed by replacing the original free bone flap, and the residual fissure and burr holes were filled with the composite scaffold with fibrin glue. In group 2 (n=2), the bone defect was closed only by replacing the original free bone flap. Three months after the treatment, computed tomography and histologic examinations were done in all animals. RESULTS: In all dogs in group 1, the defects were almost closed by new bone; whereas in group 2, the defects were filled with fibrous tissues instead of bone. CONCLUSION: This study showed that the composite scaffold made of beta-TCP, collagen, and autologous bone fragments with fibrin glue enabled reconstruction of cranial bone defects without the usual fixation materials. PMID- 16467707 TI - Intranasal zinc and anosmia: the zinc-induced anosmia syndrome. AB - OBJECTIVE: Commercial preparations of intranasal zinc gluconate gel are marketed as a remedy for the common cold. However, intranasal zinc has been reported as a cause of anosmia in humans and animals. Seventeen patients presenting with anosmia after the use of intranasal zinc gluconate are described. METHODS: The authors conducted a retrospective case series of patients presenting to a nasal dysfunction clinic and conducted complete history and physical examination on all patients, including nasal endoscopy. All patients underwent detailed odor threshold and identification testing. RESULTS: Threshold and identification testing revealed impaired olfaction in all patients. Inflammatory and traumatic causes of anosmia were excluded based on history, physical examination, and imaging. All patients diagnosed with zinc-induced anosmia or hyposmia reported sniffing deeply when applying the gel. This was followed by an immediate sensation of burning lasting minutes to hours. Loss of sense of smell was then perceived within 48 hours. Seven of 17 patients never developed symptoms of an upper respiratory infection. CONCLUSIONS: The zinc-induced anosmia syndrome, characterized by squirt, sniff, burn, and anosmia, occurs after the exposure of olfactory epithelium to zinc cation. It can be distinguished from postviral anosmia based on history. PMID- 16467708 TI - Incidence of inverted papilloma in recurrent nasal polyposis. AB - OBJECTIVES: A diagnosis of inverted papilloma in apparently normal bilateral polyps is a possible albeit rare event. The frequency of this diagnosis varies between 0.00% and 0.92%. There are no studies evaluating whether this possibility is significantly different in patients operated for recurrence when compared with those undergoing first surgery. The aim of this study is to evaluate the frequency of inverted papilloma in a series of histologic specimens obtained from patients undergoing surgery for recurrent nasal polyposis. METHODS: The authors conducted a retrospective chart and pathologic review of data from 1887 surgical interventions for removal of unremarkable bilateral nasal polyps. Frequency of inverted papilloma was assessed. RESULTS: Five cases of inverted papilloma were identified. The incidence of this diagnosis was thus 0.26% (95% confidence interval, 0.08-0.62%). Age, gender, and number of recurrences did not influence the frequency of this diagnosis. CONCLUSIONS: The occurrence of inverted papilloma in unremarkable recurrent cases of nasal polyps is rare. This rate is similar to the one observed in patients undergoing first surgery. PMID- 16467709 TI - Comparative proteomics analysis of the proteins associated with laryngeal carcinoma-related gene 1. AB - OBJECTIVES: A novel gene, laryngeal carcinoma-related gene 1 (LCRG1), had the characteristics of tumor-suppressor genes. It was cloned in our laboratory. The objective was to find and characterize the proteins related to LCRG1 and to elucidate the molecular mechanism of LCRG1. STUDY DESIGN: We used the established cell lines of Hep-2/LCRG1 (Hep-2 cells transfected by recombinant plasmid pcDNA3.1[+]/LCRG1) and Hep-2/pcDNA3.1(+) (Hep-2 cells transfected by control vector pcDNA3.1[+]) as cell models. METHODS: Two-dimensional gel electrophoresis (2-DE) technology was performed to separate the proteins of Hep-2/LCRG1 and Hep 2/pcDNA3.1(+) cell lines, respectively. The differential protein spots were analyzed by software analysis, subject to in-gel digestion, and identified by Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and electrospray ionization-quadruple time-of-flight MS/MS (ESI-Q TOF MS/MS). Then the differential expression levels of partial identified proteins were determined by Western blotting analysis and quantitative real-time reverse transcriptase-polymerase chain reaction. RESULTS: The results showed the attained 2-DE patterns of the two cell lines were well-resolved and reproducible. There were 1075+/-43 and 1027+/-23 protein spots observed in Hep-2/LCRG1 and Hep 2/pcDNA3.1(+) cell lines, respectively. The average matching rate of the two cell lines was 91%. Twenty-six differentially expressed protein spots were identified (twenty spots for MALDI-TOF-MS, six spots for ESI-Q-TOF MS/MS). Most of the characterized proteins were characterized as the members of enzymes (phosphoglycerate mutase, manganese superoxide dismutase, and so on), transcription proteins (rho gdp dissociation inhibitor), and so on. Those identified proteins might contribute to the tumor-suppressive function of LCRG1. The differential expression levels of the partial proteins were confirmed by real time polymerase chain reaction and Western blotting. CONCLUSIONS: We tentatively proposed those differentially expressed proteins were involved in the tumor suppressive process related to LCRG1. These data will be helpful to elucidate the molecular mechanism of LCRG1. PMID- 16467710 TI - Long-term result of the new endoscopic vocal fold medialization surgical technique for laryngeal palsy. AB - OBJECTIVE: The conventional surgical method for a case of unilateral laryngeal nerve paralysis with large glottal gap requires an external cervical incision. In the present study, we developed an endoscopic technique of vocal fold medialization that can make the external incision unnecessary. This procedure of autologous transplantation of fascia into the vocal fold (ATFV) was developed for the successful treatment of unilateral laryngeal nerve paralysis. However, the method seemed to be effective only for patients with a relatively mild glottal gap. STUDY DESIGN AND METHODS: In the present study, we modified the method of medialization using the ATFV technique to obtain effective closure of a large glottal gap. To overcome this difficulty, an attempt was made to extend the site of transplantation more posteriorly so as to adduct the vocal process of the arytenoid cartilage in the body of the vocal fold. RESULTS: This new technique was applied to eight cases of patients with unilateral laryngeal paralysis with severe dysphonia. None of the patients showed any evidence of falling off of the graft. Elongation of the maximum phonation time and a decrease in airflow rate during phonation were obtained with improvement in voice quality in all patients 1 year after the surgery. CONCLUSIONS: This method, with its less invasive approach, proved to be useful for the treatment of large glottal gap due to unilateral laryngeal nerve paralysis. PMID- 16467711 TI - The utility of evaluating true vocal fold motion before thyroid surgery. AB - OBJECTIVE: The objective of this study was to evaluate the utility of screening laryngoscopic examination in evaluating vocal fold (VF) mobility before thyroid surgery. METHODS: The authors conducted a retrospective chart review of 340 patients who have undergone thyroid surgery from January 1998 to June 2005 and had preoperative laryngoscopy by mirror, fiberoptic, or videostroboscopic examination. Reports of preoperative voice change or complaint and reports of preoperative VF examination, including the method of examination, were recorded. For patients with VF motion impairment, reports of the intraoperative condition of the recurrent laryngeal nerve (RLN), preoperative diagnosis based on fine needle aspiration, and final postoperative histopathologic examination results were recorded. RESULTS: Twenty-two patients were found to have preoperative VF motion impairment, of which seven (32%) patients were asymptomatic with no detectable subjective or objective voice problems. This differs significantly from the hypothesis that patients with VF motion impairment are always symptomatic (P=.009). Using voice symptoms as a screening test to predict VF motion impairment in 340 patients reveals that the sensitivity was 68%, specificity was 91%, positive predictive value (PPV) was 31%, and negative predictive value (NPV) was 98%. Among the 22 patients with preoperative VF motion impairment, five (72%) of the seven asymptomatic patients had benign, slowly progressive disease on their final histopathology reports. Six of these asymptomatic patients had their preoperative VF evaluation by fiberoptic examination, whereas one patient had indirect mirror laryngoscopy. Of 22 patients with preoperative VF motion impairment, five (22.5%) patients had abnormal VF mobility contralateral to the thyroid lesion on their preoperative evaluation, and only two of them had nerve injury reported after a previous thyroid surgery. This result differs significantly from the hypothesis that impaired mobility is ipsilateral to the side of the lesion (P=.05). CONCLUSIONS: Patients without voice complaints can have VF motion impairment. Patients can also have VF motion impairment contralateral to the thyroid lesion. Preoperative VF examination helps counsel patients appropriately about the risks of surgery and helps outline a plan for the extent of surgery while minimizing the medicolegal ramifications of iatrogenic RLN injury. PMID- 16467712 TI - Immunoglobulins and inflammatory cytokines in nasal secretions in humoral immunodeficiencies. AB - OBJECTIVE: Chronic respiratory tract infections are a common problem in patients with severe humoral immunodeficiency despite intravenous immunoglobulin therapy (IVIG), often presenting as rhinosinusitis. METHODS: Because it is unclear whether IVIG is a good substitute at the mucosal surface, we analyzed immunoglobulin levels and inflammatory cytokines (ECP, IL-8, and TNF-alpha) in nasal secretions of 13 patients with common variable immunodeficiency (CVID) and in 10 patients with IgA deficiency. RESULTS: In patients with CVID, median IgG and IgM levels did not differ significantly from controls, whereas inflammatory cytokines were markedly elevated, reflecting persistent inflammation at the mucosal site. In contrast, patients with IgA deficiency showed significantly raised IgG and IgM levels, whereas ECP and TNF-alpha were only slightly increased. CONCLUSION: Low levels of SIgA might be compensated locally at the mucosal site by high levels of IgM and IgG. Our findings implicate that adequate IVIG is not sufficient to prevent chronic inflammation of the sinuses in patients with severe humoral immunodeficiency. PMID- 16467713 TI - Polyester wax: a new embedding medium for the histopathologic study of human temporal bones. AB - BACKGROUND: Celloidin and paraffin are the two common embedding mediums used for histopathologic study of the human temporal bone by light microscopy. Although celloidin embedding permits excellent morphologic assessment, celloidin is difficult to remove, and there are significant restrictions on success with immunostaining. Embedding in paraffin allows immunostaining to be performed, but preservation of cellular detail within the membranous labyrinth is relatively poor. OBJECTIVES/HYPOTHESIS: Polyester wax is an embedding medium that has a low melting point (37 degrees C), is soluble in most organic solvents, is water tolerant, and sections easily. We hypothesized that embedding in polyester wax would permit good preservation of the morphology of the membranous labyrinth and, at the same time, allow the study of proteins by immunostaining. METHODS: Nine temporal bones from individuals aged 1 to 94 years removed 2 to 31 hours postmortem, from subjects who had no history of otologic disease, were used. The bones were fixed using 10% formalin, decalcified using EDTA, embedded in polyester wax, and serially sectioned at a thickness of 8 to 12 mum on a rotary microtome. The block and knife were cooled with frozen CO2 (dry ice) held in a funnel above the block. Sections were placed on glass slides coated with a solution of 1% fish gelatin and 1% bovine albumin, followed by staining of selected sections with hematoxylin and eosin (H&E). Immunostaining was also performed on selected sections using antibodies to 200 kD neurofilament and Na-K ATPase. RESULTS: Polyester wax-embedded sections demonstrated good preservation of cellular detail of the organ of Corti and other structures of the membranous labyrinth, as well as the surrounding otic capsule. The protocol described in this paper was reliable and consistently yielded sections of good quality. Immunostaining was successful with both antibodies. CONCLUSION: The use of polyester wax as an embedding medium for human temporal bones offers the advantage of good preservation of morphology and ease of immunostaining. We anticipate that in the future, polyester wax embedding will also permit other molecular biologic assays on temporal bone sections such as the retrieval of nucleic acids and the study of proteins using mass spectrometry-based proteomic analysis. PMID- 16467714 TI - Three-week thyroxine withdrawal: a thyroid-specific quality of life study. AB - OBJECTIVES/HYPOTHESIS: The purpose of this study is to determine the impact of a 3-week T4 withdrawal test on the quality of life (QOL) of patients undergoing investigation for residual/recurrent well-differentiated thyroid cancer. STUDY DESIGN: Prospective survey study. METHODS: 181 patients with well-differentiated thyroid cancer were surveyed using a thyroid-specific QOL-thyroid survey at three times throughout the study: prior to T4 withdrawal, after the 3-week withdrawal period, and 4 weeks after resuming T3/T4 combination therapy. RESULTS: A very small, though statistically significant, reduction was observed in the four domains of QOL defined by the QOL-thyroid survey from baseline values to 3 weeks after T4 withdrawal. The maximal difference was 2.04 (fatigue), out of a possible 10 point score, in the physical well-being category. CONCLUSIONS: The thyroid specific QOL questionnaire demonstrates a statistically significant though very small and probably not clinically significant reduction in the QOL of thyroid cancer patients undergoing an induced 3-week hypothyroid state. Three-week T4 withdrawal is a simple, cost-effective, and readily available test. PMID- 16467715 TI - Treatment of chronic posterior laryngitis with esomeprazole. AB - OBJECTIVE: To evaluate the efficacy of acid-suppressive therapy with the proton pump inhibitor esomeprazole on the signs and symptoms of chronic posterior laryngitis (CPL) in patients with suspected reflux laryngitis. STUDY DESIGN: Prospective, multicenter, randomized, parallel-group trial that compared twice daily esomeprazole 40 mg with placebo for 16 weeks. METHODS: Eligible patients had a history of one or more CPL symptoms (throat clearing, cough, globus, sore throat, or hoarseness) and laryngoscopic signs indicating reflux laryngitis based on CPL index (CPLI) scores measured during a screening laryngoscopy. Patients were randomized to treatment if their 7-day screening diary-card recordings showed a cumulative primary symptom score of 9 or higher and they had 3 or more days with moderately severe symptoms based on a 7-point scale. Efficacy was assessed by changes in symptoms as recorded by patients and investigators and by changes in CPLI scores based on laryngoscopic examinations. RESULTS: The patients' primary CPL symptom at final visit (primary efficacy end point) was resolved in 14.7% (14/95) and 16.0% (8/50) of patients in the esomeprazole and placebo groups, respectively (P=.799). Esomeprazole and placebo were not significantly different for change from baseline to the final visit in mean total CPLI (-1.66+/-2.13 vs. -2.0+/-2.55, respectively; P=.446) or any other secondary efficacy end points based on patient diary card or investigator assessments. CONCLUSION: This study provides no evidence of a therapeutic benefit of treatment with esomeprazole 40 mg twice daily for 16 weeks compared with placebo for signs and symptoms associated with CPL. PMID- 16467716 TI - Prediction of auditory brainstem reflex screening referrals in high-risk infants. AB - OBJECTIVES/HYPOTHESIS: The objectives of this study were to 1) quantify the relative importance of established risk factors for congenital hearing loss (HL), 2) identify other risk factors for congenital HL, and 3) create a prognostic system that can predict the chance of an infant having HL. METHODS: The authors conducted a retrospective cohort study with validation of a prognostic system. The authors used a medical record review on 1,863 infants admitted to level II and level III nurseries who underwent auditory brainstem reflex (ABR) hearing screening from 1998 to 1999 (derivation cohort). The primary outcome was hearing screening referral (i.e., hearing screening failure) classified as mild to moderate, moderate to severe, and severe to profound loss. To validate the prognostic system, a medical record review of a separate cohort of 437 infants admitted to a level III nursery who underwent ABR hearing screening in 2002 (validation cohort) was performed. RESULTS: In the derivation cohort, 1,513 infants (81%) passed the ABR screen at 30 dB, 77 (4%) had a unilateral referral, 243 (13%) had a bilateral referral, and 30 (1.6%) did not have a complete screening. In multivariable analysis, the following risk factors were clinically and statistically significant: craniofacial abnormalities, cytomegalovirus infection, bronchopulmonary dysplasia (BPD), maternal ethanol use, syndromes, hydrocephalus, and hyperbilirubinemia. CONCLUSIONS: Our results reaffirm the importance of several established risk factors for congenital HL, but suggest that BPD is an important predictor in high-risk populations. Whether these risk factors are causal or merely associated in congenital HL remains to be determined. PMID- 16467717 TI - Level IIb lymph node metastasis in laryngeal squamous cell carcinoma. AB - OBJECTIVES: Selective neck dissection, despite preservation of the spinal accessory nerve, can lead to some degree of postoperative shoulder dysfunction as a result of removal of level IIb lymph nodes. The aim of this study was to determine whether level IIb lymph nodes can be preserved in elective or therapeutic neck dissection as a treatment for patients with laryngeal squamous cell carcinoma (SCC). STUDY DESIGN: This was a prospective analysis of a case series. METHODS: A prospective analysis of 65 patients with laryngeal SCC who underwent surgical treatment of the primary lesion with simultaneous neck dissection from January 1999 to December 2002 was performed. During the neck dissection, the contents of the level IIb lymph nodes were dissected, labeled, and processed separately from the remainder of level II nodes and the main neck dissection specimen. The incidence of pathologic metastasis to level IIb lymph nodes and the regional recurrence within this area were evaluated. In addition, several potential risk factors for metastatic disease in the level IIb lymph nodes such as sex, age, cT stage, cN stage, and the presence of other positive lymph nodes were also evaluated. RESULTS: A total of 125 neck dissections were performed in this series. Of these dissections, 102 (82%) were elective and 23 (18%) were therapeutic. The prevalence of metastases in the level IIb lymph nodes was 1% (one of 46) and 0% (zero of 56) in clinically node-negative (N0) ipsilateral and contralateral necks, respectively, and 37% (seven of 19) and 0% (zero of four) in clinically node-positive ipsilateral and contralateral necks, respectively. There was a statistically significant association between level IIb metastases and clinically positive N stage (P<.001). The presence of other positive lymph nodes was also shown to have a statistically significant association with metastasis in the level IIb lymph nodes (P=.001). Only two of 46 patients (4%) with clinically N0 necks developed a regional recurrence. However, three of eight cases (38%) with positive pathologic level IIb lymph nodes developed regional recurrence. CONCLUSION: Level IIb lymph node pads may be preserved in elective neck dissection in patients with laryngeal SCC. However, this area should be removed thoroughly during therapeutic neck dissection in the treatment of clinically node-positive necks. PMID- 16467718 TI - Fluorodeoxyglucose-positron emission tomography/computed tomography imaging in patients with carcinoma of the larynx: diagnostic accuracy and impact on clinical management. AB - OBJECTIVES: The objectives of this study were to assess the value of F fluorodeoxyglucose (FDG) positron emission tomography/computed tomography (PET/CT) in patients with carcinoma of the larynx as compared with PET and CT alone and to assess the impact of PET/CT on further clinical management. STUDY DESIGN: This was a prospective, nonrandomized study. MATERIALS AND METHODS: Forty two patients with laryngeal cancer had 51 PET/CT examinations. There were 34 men and eight women, aged 39 to 80 years. All studies were interpreted prospectively with knowledge of the clinical history and results of previous imaging tests. The performance of different imaging modalities was compared on both a study- and lesion-based analysis for sensitivity, specificity, positive (PPV) and negative predictive value (NPV), and accuracy. Changes in patient care resulting from the PET/CT studies were recorded. RESULTS: The study analysis showed that PET/CT had a sensitivity of 92%, specificity 96%, PPV 96%, NPV 92%, and accuracy of 94% as compared with 92%, 73%, 76%, 91%, and 82% for PET, and 88%, 8%, 48%, 40%, and 51% for CT, respectively. There were 112 suspicious sites evaluated in the 51 studies. PET/CT altered management in 25 patients (59%) by sparing previously planned diagnostic procedures (n=13), by changing the planned therapeutic approach (n=9), and by guiding a biopsy to a metabolically active laryngeal area (n=3). CONCLUSIONS: The performance of PET/CT is better than standalone PET or CT in patients with cancer of the larynx. PET/CT had a major impact on management of 59% of patients. PMID- 16467719 TI - Cochlear preservation after meningitis: an animal model confirmation of adjunctive steroid therapy. AB - OBJECTIVE/HYPOTHESIS: The objective of the present study was to determine whether treating pneumococcal meningitis with a combined antibiotic and steroid regime will prevent cochlear damage, a common pneumococcal meningitis side effect. STUDY DESIGN: This was a prospective animal study. METHODS: Gerbils were randomly assigned to three experimental groups. Animals in group 1, the control animals, received intrathecal saline injections. Animals in groups 2 and 3 received intrathecal injections of Streptococcus pneumoniae to induce meningitis. Although group 2 solely was treated for 7 days with intraperitoneal penicillin injections (48,0000 units), group 3 received, in addition to the antibiotic for 4 days, 0.5 mg/kg intraperitoneal dexamethasone injections. Three months after the meningitis was induced, the animals' cochlear function was determined using auditory brainstem responses (ABRs). Fifteen frequencies were tested, five octaves at three steps per octave between 2 and 50 kHz. RESULTS: ABR thresholds were significantly elevated only in group 2. When compared with group 1, ABR thresholds were 19 dB higher (P<.05). Frequencies at the low-frequency end of the hearing range were affected more than the midfrequencies. Animals that received dexamethasone had 2-dB higher thresholds than the control group (P>.05). CONCLUSIONS: Dexamethasone therapy in conjunction with antibiotic therapy preserves cochlear function in cases of S. pneumoniae meningitis in the Mongolian gerbil model. PMID- 16467720 TI - Early laryngeal inhalation injury and its correlation with late sequelae. AB - OBJECTIVE: Inhalation injury can permanently alter normal laryngeal function. The aim of this study was to examine the early changes in voice, swallowing, and breathing in laryngeal inhalation injuries. STUDY DESIGN: This was a prospective analysis of nine patients with inhalation injuries at a tertiary care institution. METHODS: Laryngeal function of patients admitted for inhalation injury requiring intubation was documented using videostroboscopy and swallowing evaluation by the speech pathology service. Bronchoscopy was used to classify the degree of inhalation injury. Association among total body surface area, facial burns, severity of laryngotracheal injuries, and loss of function was attempted. RESULTS: All three patients with severe tracheal inhalation injury presented persistent hoarseness at 1-year follow up with abnormal videostroboscopy findings. No association was found between inhalation injury and total body surface area burned. None of the patients in this series presented permanent swallowing dysfunction. CONCLUSION: The otolaryngologist plays an important role in the initial and long-term management of inhalation injuries. Inhalation injuries should be managed in a multidisciplinary fashion. There may be a correlation between the degree of tracheal injury and laryngeal injury and hoarseness. PMID- 16467721 TI - Methicillin-resistant Staphylococcus aureus infections in acute rhinosinusitis. AB - OBJECTIVE/HYPOTHESIS: Methicillin-resistant Staphylococcus aureus (MRSA) has recently become a serious problem in various fields of medicine. However, it has rarely been studied in acute rhinosinusitis. The aim of this study was to identify the clinical manifestations and treatment outcome of community-acquired methicillin-resistant S. aureus in acute rhinosinusitis. STUDY DESIGN: This was a prospectively collected case series. METHODS: Since 2000, we have launched a prospective long-term study for bacteriology, drug susceptibility, and their changing trend in acute rhinosinusitis. Patients with the diagnosis of acute rhinosinusitis were enrolled from October 2000 through March 2003. Their middle meatus discharge was taken for aerobic culture. Antibiotic sensitivity test was performed for each isolate. RESULTS: A total of 601 patients with the diagnosis of acute rhinosinusitis were included in this study. MRSA was isolated in 16 specimens. Its prevalence rate in acute rhinosinusitis was 2.7% (16 of 601). Multiple pathogens were more frequently found in children with MRSA infection. Five of seven adults had previous nasal procedures. Eight of nine children had a history of antibiotic use. Except for two patients without follow up, the remaining 14 patients resolved after receiving oral antibiotics according to culture results. CONCLUSIONS: The incidence of MRSA infection in acute rhinosinusitis was 2.7% in our study. The most important risk factor was nasal surgeries in adults and previous antibiotic use in children. The treatment outcome of community-acquired MRSA was excellent with oral antibiotics. PMID- 16467722 TI - The histone deacetylase inhibitor sodium butyrate protects against cisplatin induced hearing loss in guinea pigs. AB - OBJECTIVE: There is a need for otoprotective agents that can be administered systemically without compromising cancer treatment. Histone deacetylase inhibitors are anticancer agents that act by upregulating the expression of cell cycle control genes. They are also neuroprotective, leading us to hypothesize that they might be otoprotective. The goal of this study was to determine if the antitumor agent sodium butyrate (a histone deacetylase inhibitor) protects against cisplatin ototoxicity when administered systemically. STUDY DESIGN: This was an animal study. METHODS: : Cisplatin was administered to guinea pigs who received either 12 days of sodium butyrate (7 d before and 5 d after cisplatin) or equivolume saline injections. Hearing was tested with distortion product otoacoustic emission (DPOAE) analysis before the start of the study and 2 weeks after cisplatin treatment. RESULTS: Guinea pigs given a single intraperitoneal injection of 14 mg/kg cisplatin experience a mean hearing loss of 8 dB across the frequencies of 3.5, 5, 7, 10, 14, and 20 kHz. Intraperitoneal injection of 1.2 mg/kg sodium butyrate per day for 7 days before and 5 days after cisplatin almost completely eliminates this threshold shift (P=.0011). CONCLUSIONS: The histone deacetylase inhibitor sodium butyrate gives almost complete protection in a single-dose model of cisplatin ototoxicity in guinea pigs. Because histone deacetylase inhibitors are anticancer agents with very few side effects, they may be candidates for clinical use during cisplatin chemotherapy. PMID- 16467724 TI - Incongruence between histologic and endoscopic diagnoses of Barrett's esophagus using transnasal esophagoscopy. AB - OBJECTIVES: The symptoms, patterns of reflux, and clinical manifestations of laryngopharyngeal reflux (LPR) differ from those of gastroesophageal reflux disease (GERD) in many ways. The purposes of this study were to determine the prevalence of Barrett's esophagus in patients with LPR using transnasal esophagoscopy (TNE) and to determine if there is agreement between TNE clinical findings and pathology results when using TNE for Barrett's screening. STUDY DESIGN: This study involved a retrospective review of the records of 200 consecutive patients with LPR undergoing esophageal screening. METHODS: The prevalence of patients with findings clinically suspicious for Barrett's and the biopsy results for those patients were reviewed. RESULTS: Of the 200 patients with LPR who were screened with TNE, 10% (20 of 200) had findings suspicious for Barrett's esophagus, and, of those, only 30% (six of 20) had biopsy-proven Barrett's metaplasia. CONCLUSION: Although TNE may be a useful screening tool for Barrett's, there is incongruence between TNE findings and biopsy results, which likely reflects suboptimal biopsy methods with TNE. New biopsy techniques such as the CDx brush biopsy may enhance the sensitivity of TNE biopsies, and future studies are needed in this area. PMID- 16467723 TI - Health-related quality of life after polypectomy with and without additional surgery. AB - OBJECTIVE: The objective of this study was to compare the health-related quality of life of patients undergoing simple polypectomy with that of patients undergoing polypectomy with additional surgery. STUDY DESIGN: This was a prospective, multicenter cohort study of adults undergoing sinonasal surgery. METHODS: Eight hundred forty-four patients received simple polypectomy and 1,004 patients received polypectomy with additional surgery. Health-related quality of life was compared at 12 and 36 months after surgery using the Sino-Nasal Outcome Test (SNOT-22). Total SNOT-22 scores may range from zero to 110 with lower scores representing better outcomes. We used linear regression to adjust postoperative SNOT-22 scores for baseline characteristics. When comparing the difference between the two surgical techniques, positive SNOT-22 scores represent a better outcome for those undergoing additional surgery. RESULTS: There were only small differences between the two groups at 12 months (difference in SNOT-22 -0.5; 95% confidence interval [CI]=-2.3-1.3; P=.58) and 36 months after surgery (difference -2.1; 95% CI=-4.4-0.2; P=.08). The additional surgery group had a slightly higher risk of excessive perioperative bleeding (8.6% vs. 6.0%; P=.04) but a slightly lower risk of revision surgery within 36 months (10.4% vs. 13.3%; P=.12). CONCLUSIONS: Nasal polypectomy with additional surgery seems to have no benefit over simple polypectomy in terms of health-related quality of life improvement for patients with nasal polyposis. PMID- 16467725 TI - Scanning electron microscopic study of the nasolabial cyst: its clinical and embryological implications. AB - OBJECTIVE: Nasolabial cyst is an uncommon midfacial cyst. It is considered to be a developmental anomaly arising from the rest of nasal respiratory epithelium. Although the cyst is a well-recognized entity, there remains some confusion of its origin, cell types, and ultrastructures. Based on the routine light microscopic study, some authors reported the epithelial cells of the inner lining of the nasolabial cyst were ciliated; some others reported they were nonciliated. To clarify this, a scanning electron microscopic study is needed. STUDY DESIGN: This was a prospective clinical series. METHODS: A transnasal marsupialization method was used to treat 10 patients with nasolabial cyst. With patients under local anesthesia, the roof of the cyst wall and a disk of nearby nasal mucosa were excised together with a sickle knife and scissors. Surgical specimens were dissected and processed for scanning electron microscopy and histochemistry. Patients were followed up for 8 to 65 months. RESULTS: Marsupialization of cysts was successfully performed on all patients. Electron microscopically, the inner surface of the nasolabial cysts in all the cases was lined with nonciliated columnar epithelium consisting chiefly of goblet cells and basal cells. It is suggested that goblet cells contributed to clear, thin, and yellow mucus present in the cyst lumen. Instead of cilia, these epithelial cell surfaces were equipped with numerous short, globular, or irregular microvilli. Apical cytoplasm of adjacent cells did not tightly adhere to each other. Instead, microsulci of 1 to 3 microm in width formed between cells. Cytoplasmic processes from the lateral border spanned the microsulcus and contacted with those from neighboring cells. CONCLUSION: The novel study has proved that the lining epithelium on the inner surface of the nasolabial cyst is columnar epithelium that chiefly consisted of two types of cells: goblet cells and basal cells. Not present were ciliated cells that were essential in the other portion of the respiratory tract. Numerous microvilli, instead of cilia, covered the inner lining of the nasolabial cyst, probably as a result of lacking the stimulation of air in ventilation as that on the other portion of the respiratory tract. The cilia of the epithelium were ill developed. PMID- 16467726 TI - The effect of erythropoietin on gentamicin-induced auditory hair cell loss. AB - OBJECTIVE/HYPOTHESIS: Mammalian auditory hair cells that are unable to regenerate and various agents, including gentamicin, can irreversibly damage the hair cells. Erythropoietin, known as the primary regulator of erythropoiesis, exerts also neuroprotective effects by binding to its receptor. We tested whether erythropoietin can protect the hair cells from gentamicin-induced damage. STUDY DESIGN: This study localized the erythropoietin receptor in the cochlea and analyzed the effect of erythropoietin on gentamicin-damaged hair cells in vitro. METHODS: Expression of erythropoietin receptor in the rat cochlea was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry. Protection of auditory hair cells from gentamicin was tested in vitro by exposing cultured rat organs of Corti with increasing concentrations of erythropoietin (0.1 U/mL, 1 U/mL, and 10 U/mL). RESULTS: We detected erythropoietin and erythropoietin receptor mRNA expression in the organ of Corti, spiral ganglion, and stria vascularis by RT-PCR. Immunohistochemistry revealed that the erythropoietin receptor localizes to the outer and inner hair cells and supporting cells of the organ of Corti, as well as to the spiral ganglion cells and the stria vascularis. Significantly less hair cell loss occurred in the organs of Corti that were pretreated with 0.1 U/mL erythropoietin as compared with samples treated with gentamicin only. CONCLUSION: Decreased hair cell loss in erythropoietin-treated organs of Corti that had been exposed to gentamicin provides evidence for a protective effect of erythropoietin in aminoglycoside-induced hair cell death. PMID- 16467727 TI - Auditory responses in cochlear implant users with and without GJB2 deafness. AB - OBJECTIVE/HYPOTHESIS: It is reasonable to suppose that the pattern of sensorineural damage along the length of the cochlea depends on the etiology of a hearing loss (HL). In GJB2-related deafness, we hypothesize that gap junction deficits are uniformly distributed and will result in similar damage along the length of the cochlea as compared with non-GJB2 subjects. We assessed this by measuring patterns of neural activity and hearing from apical versus basal cochlear implant electrode regions. STUDY DESIGN: This was a prospective, blind, controlled study. METHODS: Blood from 301 pediatric cochlear implant users was analyzed for mutations in GJB2 by direct sequencing. After exclusion of patients with monoallelic GJB2 mutations, associated syndromes, or risk factors for HL that were not congenital, 39 children with biallelic GJB2 mutations and 58 without GJB2 mutations were evaluated. Hearing was measured before implantation at frequencies ranging from 250 Hz to 8 kHz. After implantation, neural activity at the apical and basal ends of the implanted array was measured using electrically evoked compound action potentials of the auditory nerve (ECAPs) and evoked stapedius reflexes (ESRs). RESULTS: GJB2 and non-GJB2 groups were not significantly different with respect to sex, age at implantation, duration of auditory deprivation, hearing aid use, duration of aided hearing, ear implanted, implant model, or depth of insertion (P>.05). Children with GJB2-related HL had greater similarities between low- and high-frequency residual hearing and between neural activity electrically evoked at apical and basal regions of the cochlea as compared with children with non-GJB2-related HL who demonstrated larger deficits in basal regions. CONCLUSION: Results suggest more consistent spiral ganglion survival along the length of the cochlea in GJB2-related HL as compared with non GJB2-related HL, which appears to involve a decreasing gradient of spiral ganglion survival from the apex to the base of the cochlea. Our findings support our premise that in GJB2-related HL, dysfunction of gap junctions likely occurs to a similar degree in the apical and basal regions of the cochlea. This knowledge might be used to customize implantable devices for patients with HL in the future. PMID- 16467728 TI - Upregulation of surfactant protein A in chronic rhinosinusitis. AB - OBJECTIVES: Surfactant protein A (SP-A) is protein that appears to play an important role in mammalian first-line host defense. However, the presence of SP A in the human paranasal sinus mucosa is not well known. The purpose of this study was to investigate the expression of SP-A protein in human paranasal sinus mucosa and to compare the expression of SP-A mRNA between normal paranasal sinus mucosa and paranasal sinus mucosa with chronic rhinosinusitis. METHODS: Paranasal sinus mucosa samples from 10 patients who underwent surgery for chronic rhinosinusitis without polyps and 10 normal control subjects were used. Reverse transcriptase polymerase chain reaction was done to detect SP-A mRNA. The expression level of SP-A transcripts was semiquantified with desitometry. Cellular localization of SP-A was sought by using immunohistochemistry. RESULTS: SP-A mRNA and protein were expressed in the human paranasal sinus mucosa. SP A/GAPDH mRNA ratio in the paranasal sinus mucosa with chronic rhinosinusitis was greater compared with that in normal paranasal sinus mucosa (P<.05). Immunohistochemical staining revealed SP-A immunoreactivity in the epithelial cells and submucosal glands of paranasal sinus mucosa in both control subjects and chronic sinusitis patients. Stronger immunoreactivity was observed in chronic rhinosinusitis mucosa as compared with normal paranasal sinus mucosa. CONCLUSION: SP-A mRNA and protein are present in both normal and diseased human paranasal sinus mucosa. These results may provide potential targets for novel therapy of chronic rhinosinusitis. PMID- 16467730 TI - Searching the literature using medical subject headings versus text word with PubMed. AB - OBJECTIVE/HYPOTHESIS: This study was conducted to investigate the performance of two search strategies in the retrieval of information from the National Library of Medicine (NLM) on otolaryngology-head and neck surgery related conditions and diagnoses using PubMed. METHODS: Two search strategies-one based on the use of Medical Subject Headings (MeSH) and the second based on text word searching-were compared. RESULTS: The MeSH search provided a more efficient search than the text word search. CONCLUSIONS: Head and neck surgeons can most efficiently search the NLM using PubMed as a search engine by initiating the search with MeSH terms. Once a key article is identified, the searcher should use the "Related Articles" feature. PMID- 16467729 TI - Safety and efficacy of nasal application of CpG oligodeoxynucleotide as a mucosal adjuvant. AB - OBJECTIVES: Nasal vaccination is an effective regimen to prevent upper respiratory infections. An appropriate adjuvant is required for the development of a nasal vaccine. The safety and efficacy of CpG oligodeoxynucleotide (ODN) as a mucosal adjuvant was examined. METHODS: Mice were nasally administered various doses of CpG ODN weekly, a total of three times. Histologic changes in the spleen and the nasal mucosa were examined, and the alterations in cell subpopulations were analyzed by flow cytometry. In addition, the mice were nasally immunized with P6 outer membrane protein of nontypeable Haemophilus influenzae (NTHi) and CpG ODN, and P6-specific immune responses were examined. RESULTS: No inflammation or tissue damage was observed locally or systemically after nasal administration, even with a high dose of CpG ODN. A high dose of CpG ODN induced an increase in CD8+ T cells in the nasal mucosa and B cells in the spleen. When CpG ODN was coadministered with P6, P6-specific mucosal and systemic immune responses were effectively induced, since high levels of the specific IgA and IgG were detected in the nasal wash and serum, respectively. CONCLUSIONS: These findings suggest that CpG ODN is a safe and effective mucosal adjuvant. Further, nasal vaccination with P6 and CpG ODN might be an effective regimen to prevent upper respiratory infections. PMID- 16467731 TI - Stridor and dysphagia in diffuse idiopathic skeletal hyperostosis (DISH). AB - As otolaryngologists, we are the first consulted for stridor and dysphagia. One must consider both extrinsic and intrinsic etiologies in the differential diagnosis of these symptoms. We report a series of patients with diffuse idiopathic skeletal hyperostosis (DISH) who presented with stridor or dysphagia. We describe the initial presenting symptoms, physical examination/radiographic findings, and discuss the management options. Traditional teaching is that surgery is rarely indicated for DISH of the cervical spine. Recommendations regarding the role of surgery as well as a review of our surgical experience are discussed. PMID- 16467732 TI - Surgical improvement of hypotonicity in tracheoesophageal speech. PMID- 16467733 TI - Treatment of community-associated MRSA infections. PMID- 16467734 TI - Nelarabine (Arranon) for T-cell acute lymphoblastic leukemia. PMID- 16467735 TI - Phosphate binders. PMID- 16467736 TI - Treatment of insomnia. PMID- 16467737 TI - Capturing circulating endothelial progenitor cells: a new concept tested in the HEALING studies. PMID- 16467738 TI - Current drug-eluting stents in complex patients and lesions. AB - Drug-eluting stents (DES) are playing an increasingly important role in the treatment of coronary artery disease. These new devices work by releasing controlled amounts of pharmacological agents with anti-restenosis properties at the implantation site. Most of them use polymer coating as a drug carrier, but concerns about long-term negative effects of a permanent polymer coating have stimulated the development of non-polymer DES or DES based on bioabsorbable polymers. Several randomized studies with DES have demonstrated their superiority over bare metal stents mostly in selected patients and lesion subsets. Accumulating evidence is showing significant differences in performance between currently used DES. These differences are more pronounced in complex, high-risk subsets of patients and lesions and should be considered during the process of DES selection for the individual patient. Interventional cardiologists have learned that patients who receive DES require a more prolonged antiplatelet therapy, but the optimal length and regimen are still unclear and further investigations are needed. Major advances in interventional cardiology have caused a dramatic shift away from aorto-coronary bypass surgery and an increase in the complexity of percutaneous coronary interventions. Observational and specifically designed randomized studies are currently addressing the issue of the role of DES in complex situations including in-stent restenosis, ostial and bifurcation lesions, chronic occlusions, small vessels, long lesions, saphenous vein grafts, multivessel disease, left main disease, acute myocardial infarction and diabetes mellitus. Although definitive answers are still to come from ongoing research, available data support the use of DES in most of these situations. PMID- 16467739 TI - Percutaneous valve replacement: weird or wonderful? AB - Percutaneous treatment of valve diseases has proven to be an effective alternative to open surgery since the early 1980's. For most patients with rheumatic mitral stenosis, use of percutaneous catheter-based techniques as balloon valvuloplasty for the treatment of both congenital pulmonary stenosis as well as trans-septal commissurotomy has become a good therapeutic option. Furthermore in the mid 1990's in vitro studies demonstrated the feasibility of percutaneous catheter-based valve implantation first in pulmonary and subsequently in aortic position. Afterwards initial in vivo implantations were successfully undertaken. Limitations of this new branch of transcatheter-based techniques are: risk of embolization due to pre-dilatation; difficult deployment; migration of valved stent; paravalvular leakage and limited durability of the implanted valve. Hence, many advances and improvements are necessary prior to pronouncing a new real alternative and safe therapeutic option. Up to now the gold standard for the treatment of heart valve disease is still open surgical valve repair and replacement. Percutaneous valve replacement procedures offer substantial advantages both to patients and medical care providers. From a medical point of view, they may help reduce surgical risks, offer a less invasive procedure, lower complication rates and shorten rehabilitation times in future. From an economic standpoint, shorter hospital stays result in overall cost reduction. PMID- 16467740 TI - Percutaneous transcatheter closure of patent foramen ovale. AB - In presence of a patent foramen ovale (PFO) with cryptogenic cerebral embolism, traditional therapy consists of oral anticoagulation or antiplatelet therapy. Surgery was considered only in case of recurrence. Transcatheter closure of PFO is currently performed. The availability of new user friendly devices and the increasing knowledge of pathophysiology, epidemiology, and follow-up of these patients has broadened the indications and marked reduced morbidity related to interventional PFO closure. This review presents the current knowledge and our own data concerning transcatheter closure of PFO. PMID- 16467741 TI - Noninvasive assessment of coronary stenosis. AB - Invasive coronary angiography (ICA) has been the gold standard in the diagnosis of coronary artery disease (CAD) for a long time and angiographic procedures performed annually in the United States have steadily increased during the last 25 years. The unmatched temporal and spatial imaging resolution, the high level contrast between the coronary lumen and the adjacent structures, and the ability to concurrently perform percutaneous coronary interventions (PCI) are the technical advantages of coronary angiography. However, the isolated intraluminal imaging, the relatively high financial cost of the procedure, and the risks associated with catheterization may be limitations to its use. Moreover, a better selection of patients really in need of ICA and PCI is necessary since many patients referred for ICA are found not to have significant CAD. Therefore, a remarkable technical progress has been achieved by both cardiac magnetic resonance and cardiac computed tomography in this regard for the noninvasive detection of coronary stenoses. PMID- 16467742 TI - Ultrasound vascular screening for cardiovascular risk assessment. Why, when and how? AB - Carotid artery vessel wall assessment in the form of intima-media thickness (IMT) has been identified since the late 1970's as a sensitive tool to detect atherosclerosis, predict its sequelae and detect its progression and regression. Unfortunately the technique has remained confined to large multicenter clinical research trials and no consensus has been developed regarding methodology, analysis and interpretation and no agreed upon clinical protocol that could be used in clinical practice exists. The need for an accepted clinical protocol has become acute especially since the technique has been recommended by writing groups such as American Heart Association as a useful tool for risk stratification in those with unclear or intermediate risk of cardiovascular (CV) disease. The advent of automated edge detection software and a reimbursement of this technique by insurers make it compelling that clinical consensus is reached soon. Accurate data collection methodology and measurement precision are essential; as such a method that is sensitive yet not cumbersome is required for clinical utility. This review will give a short introduction to the studies that confirm value of IMT in detecting atherosclerosis and predicting its sequelae, followed by a discussion on the appropriate clinical method of imaging and reporting. Other controversial areas in methodology such as difference between plaque vs IMT in CV risk prediction will be discussed. Finally tools and skill a clinician will need to be able to do this technique will be discussed. PMID- 16467743 TI - Carotid artery stenting--current status and future directions. AB - Carotid endarterectomy has been the standard of therapy for carotid occlusive disease in stroke prevention. More recently, carotid angioplasty and stenting became an important alternative in the treatment of carotid occlusive disease. The widespread use of cerebral protection devices has substantially decreased the morbidity of the procedure. As the experience with carotid stenting increases, so does the enthusiasm for the potential of this minimally invasive carotid intervention to become the main treatment option for the high risk patients, as well as for the average asymptomatic patient. Although current available data indicate that the results regarding success and complication rates are similar between carotid stenting and endarterectomy, several prospective clinical trials are currently in progress to evaluate the efficacy of carotid artery stenting in different patient populations and definitively establish its role in the treatment of carotid disease. This paper reviews the current status of carotid stenting, including results from clinical trials, technical aspects and controversial issues and strategies to provide cerebral protection from embolization. PMID- 16467744 TI - Percutaneous vascular interventions in the superficial femoral artery. A review. AB - The superficial femoral artery (SFA) is a frequent target of atherosclerotic disease predominantly in the proximal section near the bifurcation to the deep femoral artery and in the distal section where the adductor muscles tend to compress the artery. In the past, SFA revascularization was the domain of vascular surgery (femoropopliteal and femorodistal bypasses). However, with the development of endovascular treatment and advancing techniques as well as more sophisticated stenting material and balloons, endovascular treatment is nowadays not just a treatment option but, in most cases, preferable at least as initial revascularization procedure in the treatment of peripheral artery vascular disease. In the last years, many efforts have been made to fight restenosis in revascularized artery segments after stenting and/or angioplasty. This article aims to give a review on this topic including the most recent experience with the various latest revascularization techniques such as drug eluting stents, coated stent grafts, brachytherapy, cryoplasty, cutting balloons, and drug coated balloons. PMID- 16467745 TI - Percutaneous vascular interventions in renal artery diseases. AB - Renal artery stenosis (RAS) is a progressive manifestation of atherosclerosis. It is associated with hypertension and progressive renal failure. Noninvasive testing includes renal artery duplex, computed tomographic angiography (CTA) and magnetic resonance angiography (MRA). Percutaneous transluminal renal angioplasty and stenting (PTRAS) is indicated for significant atherosclerotic RAS while percutaneous transluminal renal angioplasty (PTRA) is indicated for fibromuscular dysplasias (FMD) associated with the proper clinical indications. PTRAS is associated with a high technical success rate and an acceptable adverse event and restenosis rate. PTRAS appears to improve control of hypertension and renal preservation. All patients should be followed clinically and with periodic duplex ultrasonography. Restenosis is treated with repeat angioplasty and occasionally stenting. Current and future areas of investigation will involve distal protection and drug eluting stents. PMID- 16467746 TI - Contrast-induced nephropathy: epidemiology and prevention. AB - Contrast-induced nephropathy (CIN) is a leading cause of iatrogenic acute kidney failure. Periprocedural CIN results in a greater risk of requiring renal replacement therapy, prolonged hospitalization, excessive health care costs, potential long term kidney impairment and mortality. Identified risk factors for CIN include premorbid chronic kidney disease, diabetes mellitus, congestive heart failure, critical illness and volume of administered contrast media. Prophylactic interventions for the prevention of CIN remain controversial and uncertain. In this review we critically appraise the evidence for prevention of CIN. In general, every attempt should be made to correct underlying volume depletion, discontinue potential nephrotoxins, reverse any acute kidney dysfunction or when not possible, consider delay of procedure or an alternative modality for imaging. A minimum volume of contrast media should be employed, including going left ventriculogram and performing staged procedures if applicable. There are few interventions with quality evidence for reducing the incidence of CIN. procedure hydration and the use of nonionic iso-osmolar contrast media have consistently demonstrated efficacy. For patients at high risk, there is evidence to suggest benefit with N-acetylcysteine. Clinical studies with adenosine antagonists are encouraging; however, further confirmatory trials are required. Based on the available studies, there is inadequate evidence for the routine use of hemofiltration, atrial natriuretic peptides, calcium channel blockers, or prostaglandins. There is no evidence to support prophylaxis with diuretic therapy, forced diuresis, low dose dopamine, fenoldopam, captopril, or endothelin receptor antagonists. Despite recent advances in the epidemiology, pathophysiology and natural history of CIN, few effective prophylactic or therapeutic interventions have conclusively demonstrated evidence for a reduction in CIN incidence and no therapy has proven efficacious once CIN is established. PMID- 16467747 TI - Subcutaneous enoxaparin following thrombolysis and intravenous unfractionated heparin in ST-elevation acute myocardial infarction: safety and efficacy of low vs full dose. AB - AIM: In ST-segment elevation myocardial infarction (STEMI) treated with fibrin specific thrombolytic agents, early intravenous unfractionated heparin (UFH) is warranted. Low molecular weight heparin Enoxaparin currently represents an alternative to UFH, to be used until hospital discharge. Since optimal dosing of subcutaneous Enoxaparin is not standardized, we conducted an observational study to compare safety and efficacy of low (4,000 U once daily) vs full dose (100 U/kg twice daily) regimens. METHODS: All STEMI patients successfully treated with tenecteplase and intravenous UFH and referred to the Catheterization Laboratory between June 2002-November 2003 for predischarge coronary angiography, were evaluated. The primary end-point was the composite of hemorrhages and residual angina/reinfarction during Enoxaparin administration, whereas secondary end points were occurrence of venous thromboembolism (VTE) during Enoxaparin administration, and infarct-related artery (IRA) patency rate at predischarge coronary angiography. RESULTS: Out of 123 patients, 57 (M/F 45/12, mean age 65.8+/-8.1 years) received low dose, and 66 (men/women 45/21, mean age 62.6+/ 11.8 years) full dose subcutaneous Enoxaparin. The incidence of the composite primary end-point was comparable in both groups (19% vs 26%; P=NS). Also, null was the occurrence of VTE, whereas the IRA patency rate did not significantly differ in the 2 groups (84% vs 86% TIMI 3 and 11% vs 9% TIMI 2 flow grades; P=NS). CONCLUSIONS: In patients with STEMI undergoing successful recanalization with tenecteplase and intravenous UFH, low dose subcutaneous Enoxaparin appears preferable to full dose, in the light of comparable safety and clinical efficacy and superior easiness of use. PMID- 16467748 TI - Diagnosis and outcome of renal function in patients with renal artery stenosis: which role have color Doppler sonography and magnetic resonance angiography? AB - AIM: Minimally invasive diagnostic techniques would be useful in the preoperative diagnosis of patients with hypertension and ischemic renal disease. The aim of our study was to compare color Doppler sonography (CDS), and magnetic resonance angiography (MRA) with the reference standard, digital subtraction angiography (DSA), in the diagnosis of renal artery stenosis. METHODS: Thirty-nine patients with arterial hypertension and monolateral or bilateral renal artery stenosis documented by CDS underwent renal artery MRA and then DSA during corrective percutaneous transluminal angioplasty. CDS and MRA scans were evaluated by 3 independent observers who studied 78 main renal arteries. Stenosis of 70% or more were regarded as significant. Sensitivity, specificity, positive and negative predictive values and two-sided 95% confidence intervals of CDS and MRA for the detection of significant renal artery stenosis were calculated. The statistical significance of the differences in sensitivities between CDS and MRA was assessed by means of the kappa test (< or =1). RESULTS: CDS and MRA, therefore, both achieved 97.6% sensitivity and 100% specificity for diagnosing stenoses at the origin of the renal artery; CDS yielded 100% sensitivity and 97.1% specificity and MRA 87.5% sensitivity and 98.6% specificity for diagnosing stenosis in the intermediate distal segments. CONCLUSIONS: Statistically significant differences between CDS and MRA in the diagnosis of renal artery stenosis have not been observed. However, according to our experience, CDS is the preferred technique because it also provides useful information about the development of kidney disease before correction. PMID- 16467749 TI - Predictors of renal artery stenosis in patients with normal renal function undergoing coronary angiography. AB - AIM: A number of patients with normal renal function undergoing coronary angiography have shown a renal artery stenosis (RAS). Detection of unknown RAS may influence therapeutic strategy in patients with coronary artery disease (CAD) candidate to coronary revascularization. Prevalence of RAS in patients with normal renal function has not been yet fully investigated. We retrospectively evaluated the prevalence of RAS in patients with normal renal function undergoing coronary angiography and candidate to coronary revascularization. METHODS: Medical records of consecutive patients underwent coronary angiography at a single public institutions over a twelve-month period were evaluated. Patients with normal renal function undergoing coincident diagnostic renal angiography to evaluate renal vessels on the basis of clinical criteria and at least one-vessel CAD were analyzed. Moderate to severe arterial stenosis (>50% stenosis), vessel occlusion were noted as significant angiographic findings. RESULTS: Angiographically significant RAS were reported in 35 (17%) of 205 consecutive patients (mean age 67.1+/-12.8 years, mean serum creatinine 0.8+/-0.5 mg/dL, mean glomerular filtration rate 112+/-13 mL/min). Twenty patients (9.8% of total) underwent renal angioplasty and stenting before successful coronary revascularization. Multivariate logistic regression analyses revealed three vessel CAD (odds ratio[OR] 8.71; 95% confidence interval [CI] 2.24-40.8; P=0.002), hypertension (OR 2.34 CI 95% 0.96-6.9; P=0.048), and hypercholesterolemia (OR 2.851; CI 95% 1.03 to 7.9; P=0.044) as independent predictors of RAS. CONCLUSIONS: The association of significant RAS with CAD is relatively high in patients with normal renal function. Renal semi-selective or selective angiography may contribute to detect unknown significant RAS in patients undergoing coronary angiography: our small series suggests that this strategy may be useful also in patients with normal renal function in presence of three- or four-vessel CAD and multiple risk factors. PMID- 16467750 TI - [Cardiovascular disease in early collagen diseases]. AB - AIM: The purpose of this paper was to verify whether there is any sign of involvement of the cardiovascular system in the early stages of collagen diseases. METHODS: Seventeen patients (10 female and 7 male, average age 41.35 +/ 9.85 years) (group A) recruited at the Ambulatory of Internal Medicine for suspected collagen diseases with period of onset of the symptomatology less than 6 months, were analyzed. Ten patients were excluded from the study: 8 had been suffering from systemic lupus erythematosus (SLE) for a number of years, 2 were older than 80 and were suffering from concomitant pathologies (diabetes mellitus and hypertension) which would have invalidated the evaluation of valvular changes like thickening. The patients were followed up for 2 years. Clinical diagnosis was made in many cases many months after the observation using the criteria of the American Rheumatic Association (ARA). All patients were subjected to titration of the following autoantibodies by means of the immuno-fluorimetry method: ANA, anti-ENA (SSA, SSB, SM, SM-RNP, SCL-70, Jo-1), anti-nDNA, anti histones. The cardiological evaluation was carried out by echography (Cardioline 12 leads) and echocardiographic examination (Aloka 2000 and HP sonos 5500 with 2.5 and 3.5 MHz probe) looking for thickening of both valvular flaps (> 3 mm for the mitral and > 2 mm for the aorta), myocardial involvement by studying global and regional kinesis of the left ventricle; pericardial involvement. The control group consisted of 17 healthy subjects with the same sex and age distribution (10 male, 7 female, average age 40.35 +/- 9.80 years) (group B). RESULTS: Eleven patients (64%) proved to be suffering from SLE, 3 (17%) from mixed collagen diseases (MC), 3 (17%) from systemic sclerosis (SS). Cardiac anomalies were observed in 12 patients: in 3 (17%) mitral valve thickening was observed (2 with SLE, 1 with SS), in 2 (11%) thickening associated with mitral valve insufficiency (with MC), in 1 (5%) isolated mitral valve insufficiency (with SLE), in 1 (5%) thickening and slight aortic insufficiency (with SLE), in 1 (5%) mitral valve vegetations (with SLE), in 2 (11%) pericardial effusion (with SLE), in 2 (11%) diastolic changes (with SS). The parameters relative to wall thickness between the 2 groups showed statistically significant differences (mitral 3.1 +/- 0.7 vs 2.3 +/- 0.4 P = 0.0005; aorta 1.7 +/- 0.2 vs 1.5 +/- 0.3 P = 0.03). CONCLUSIONS: In patients observed in the early stages of collagen diseases, cardiac involvement was observed in 70% of cases, but the data require confirmation in a larger sample. The authors, however, believe that the early identification of such involvement is useful from both the diagnostic point of view and from the point of view of patient treatment. PMID- 16467751 TI - [Role of Gated-SPECT in the early diagnosis of ischaemic cardiopathy in the diabetic patient]. AB - Diabetes presents a higher risk of both symptomatic and asymptomatic coronary artery disease (CAD). Myocardial perfusion SPECT, particularly associated with Gated acquisition, makes it possible to diagnose CAD on the basis of the presence and extent of perfusion defects, and to carry out the stratification of the cardiological risk (infarction or cardiac death) by analyzing the results of perfusion and of the functional data (ejection fraction, EF, and wall movement). Gated myocardial SPECT in the diagnosis and stratification of risk has proved superior to clinical evaluation or the exercise test alone, and it is also superior to echostress/dobutamine in the evaluation of monovasal disease. These data have been confirmed not only in the population at large but also in the diabetic population, making the technique important also in such patients also affected by coronary disease which is earlier and more advanced in diagnosis. Scintigraphic data make it possible to classify patients into various risk categories (low, intermediate and high) on the basis of which a specific therapeutic approach can be established. The present review develops the above concepts, taking into consideration the consolidated studies present in the literature, for the purpose of pinpointing the diagnostic approach in which exercise or SPECT myocardium-scintigraphy can be assigned. PMID- 16467753 TI - Male infertility: an emerging issue. PMID- 16467752 TI - Giant coronary artery aneurysm in association with systemic arterial ectasia. A case report. AB - Since the advent of coronary angiography, coronary artery aneurysm has been diagnosed with increased frequency. The etiology of coronary artery aneurysm is atherosclerosis in 50%, followed by other causes. In a 71-year-old man with previously documented abdominal aortic aneurysm of 6 cm diameter and ectasia of both left and right middle cerebral arteries, thoracic magnetic resonance imaging (MRI) demonstrated a large hollow para-cardiac mass (maximum diameter of 7 cm) lying in the anterior-lateral part of the atrio-ventricular sulcus. Coronary arteriography confirmed the aneurysmatic nature of the proximal tract of left anterior descending (LAD) artery lesion. Screening for laboratory signs of vasculitis was negative and other vascular and systemic diseases were excluded, suggesting an atherosclerotic aetiology of the aneurysm. In the absence of current cardiac symptoms, conservative management has been chosen and the patient is still well 2 years after presentation. PMID- 16467754 TI - Extracranial complications of chronic suppurative otitis media. AB - The aim of this study was to find out the pattern of extracranial complications of CSOM cases who attended to the department of ENTD, Mymensingh Medical College Hospital during the period from July'1999 to June' 2001. Different types of extracranial complications of CSOM were presented here. A total of 100 cases, diagnosed clinically and radiologically were included in the study of which 66 were male and 34 were female giving a male to female ratio of 1.94 : 1 (p < 0.05). Majority cases (53) were in the age group of 11- 20 years followed by the age group of below 10 years where there were 30 cases. Majority (64) cases came from low socio-economic class. The number of different types of extracranial complications of CSOM were as follows: mastoid abscess 57, discharging sinuses 28, purulent labyrinthitis 07, Bezold's abscess 04, fascial nerve paralysis 03 and zygomatic abscess 01. In all cases of CSOM Cholesteatoma were detected. In 14 cases, Cholesteatoma were associated with granulation tissue/polyp. Modified radical mastoidectomy were performed in all cases. On analyzing the findings of the present study it was observed that mastoid abscess were the predominant extracranial complications of CSOM, affecting mostly the male population of 11 - 20 years age group coming from low socio-economic class. PMID- 16467755 TI - Gross morphometry of human placenta in eclampsia. AB - The study was done to see the gross morphologic changes of placenta in human normal pregnancy and eclapmsia, with the aim that it would be able to increase the general body of knowledge regarding the gross morphology of normal placenta and the changes that occur in eclampsia in our population. Total of 45 (n=45) placenta, 25 (n=25) from eclampsia and 20 (n=20) from normal pregnant cases were collected from Gynaecology and Obstetric Department of Mymensingh Medical College & Hospital (MMCH). Study was done in Anatomy Department of Mymensingh Medical College (MMC). Macroscopic study of the formol saline fixed placentas revealed that, compared to the controls there was trends of lower weights and volumes of placentas in eclamptic group. Statistical significance of difference between two groups was calculated by using Students "t" test. A difference between the two groups was considered to be significant when p<0.005. PMID- 16467756 TI - Pregnancy outcome in adolescent and adult - a case comparison study. AB - This study outlined the consequences of adolescent pregnancy with comparison to adults in Mymensingh Medical College Hospital, Mymensingh. It was a case comparison study. The study population was the mothers who admitted and delivered at the Obstetrics and Gynaecology ward of hospital during April-June 2003. Sample size was 220 mothers, out of which 110 were primigravidae adolescent mothers and 110 were adults. The study finding showed that adolescents were 2.105 times and 3.679 times more at risk than adults to suffer from anaemia and UTI during pregnancy. Among adolescents 74(67.3%) and in adults 39(35.5%) had obstetric complications during pregnancy and the adolescents were 3.742 times more at risk than the adults. PET and Eclampsia were more common in adolescent than adult mothers. 86(78.2%) adolescents and 36(32.7%) adult mothers had complications before delivery. Pre term labour, Premature rupture of membrane (PROM) and Intra uterine foetal death (IUD) were more common in adolescents than adult mothers. Stillbirths were more in adolescent mothers 23(20.9%) and adults 11(10.0%). There was more low birth weight (LBW) newborns in adolescents 42(38.2%) than adults 18(16.4%). As a result the adolescents were 3.157 times more likely to give birth to low birth weight babies . The newborns of the adolescent mothers were nearly 2 times more at risk to develop asphyxia neonatorum (p< 0.030, odds ratio=1.806). PMID- 16467757 TI - Alteration in iron status in pre eclampsia. AB - The aim of the study is to compare and contrast serum iron status in pre eclamptic women with normal pregnant women which may help in the establishment of diagnosis of pre eclampsia before appearance of its clinical manifestation. A total of 82 women in the last half of pregnancy, between 17 to 40 years of age, who attended the model family planning clinic, out patient and in patient departments of Obstetrics and Gynecology unit of Mymensingh Medical College Hospital, Mymensingh were selected for this purpose before any treatment was given in present pregnancy. Out of them 32 pregnant women were taken as control because they did not show any evidence of complication during the time of selection and 50 pregnant women were randomly selected as cases on the basic of having pre eclampsia. Mean value of serum iron was significantly increased in the pre eclamptic women in comparison to controls whereas mean values of both total iron binding capacity (TIBC) and unsaturated iron binding capacity (UIBC) were significantly decreased in pre eclamptic women in contrast to controls. The results allude to the possible contribution of released iron free radicals from ischaemic placenta in pre eclampsia to its etiology. So, routine investigation of serum iron status of pregnant women as part of antenatal checkup may help in the establishment of diagnosis of pre eclampsia before appearance of its clinical manifestation. PMID- 16467758 TI - Serum copper in rural women taking combined oral contraceptive. AB - This prospective case-control study included 78 women between 15 to 45 years of age from rural area to see changes in serum copper level as a consequence of oral contraceptive use. Among the subjects, 34 women were included as controls because of not taking any form of hormonal contraceptives neither during the time of selection nor during one-year period prior to the study. Women in the control group were motivated to consume oral pill (Sukhi) for 3 consecutive cycles. At the 3(rd) month, 25 such women became available and henceforth included as cases on longitudinal basis. Another 44 women were randomly selected as cases on the basis of using combined oral contraceptives (Sukhi) for a duration of 4 months onwards. Considering different duration of oral contraceptive (OC) use, subjects were grouped as follows: Group I (n=34)--> controls, Group II (n=25)--> 3 months, Group III (n=17)--> 4 months - 2 years and Group IV (n=27)--> >2 years. Finally, 103 samples of blood (34 from controls and 69 from oral contraceptives users) were collected for estimation of Serum Copper (mgm/dl) by Atomic Absorption Spectrometry using UNICAM-AA Spectrometer. Mean+/-SD of Serum Copper significantly increased in all 3 contraceptive groups in comparison to controls (p<0.001). Further study including larger population from rural area was recommended to see correlation among serum copper and other trace elements with side effects of hormonal contraceptives. This preliminary study tried to explore the possibility of establishing biochemical monitoring of serum trace elements in OC users. PMID- 16467759 TI - Pattern of change of weight following birth in the early neonatal period. AB - Birth weight of new born will vary according to gestational age, maternal, Placental and foetal intrauterine conditions. Postnatal weight change in the first few days of life (early neonatal period) may vary according to gestational age and birth weight. This may create problem in management of these babies. Birth weight and their pattern of change were followed on 105 newborns. Questionnaire and observation charts were the research Instruments and statistical analysis were done by using manual calculator. It was observed that full term newborns had better control on initial weight change following birth than preterm and low birth weight babies. Majority of term newborns loss their weight up to 5 days of life and regained their initial weight by 10 days. Maximum weight loss for longer duration occurred in preterm and very low birth weight babies who regained their initial weight by 10 - 14 days and some required more than 21 days. PMID- 16467760 TI - Comparison between ELISA and ICT- MycoDot in adult pulmonary tuberculosis. AB - The present study was conducted to see the diagnostic efficacy of serum anti-TB IgG antibody detection in adult pulmonary tuberculosis (TB) by ELISA and Immunochromatography test (ICT) method using commercially available diagnostic kit (Pathozyme TB complex and MycoDot test respectively). This case-control study included 48 cases of adult pulmonary TB within the age range of 15-65 years. Among them, 22 (45.9%) were AFB positive confirmed cases, 12 (25.0%) were AFB negative clinically diagnosed cases before treatment and 14 (29.1%) were clinically diagnosed cases on treatment. Age and sex matched 33 controls were also included, of which 19 (57.6%) were healthy and 14 (42.4%) were sick with non specific pulmonary infection. Overall sensitivity and specificity of ELISA were 60.42% & 96.97% (p<0.001) and those of MycoDot as 72.92% & 93.94% (p<0.001). Rate of both ELISA and MycoDot positive were higher among clinically diagnosed cases on treatment with respective values as 71.42% and 85.71%. Respective values among AFB positive confirmed TB cases were 68.18% and 72.72%. As, both ELISA and MycoDot yielded be used as a confirmatory test for TB. PMID- 16467761 TI - Intravesical adjuvant therapy using mitomycin C. AB - Bladder cancer is mostly superficial at first diagnosis. High incidence of recurrence is the major problem after initial management with transurethral resection (TUR) of bladder tumor. Adjuvant chemotherapy has been advocated to reduce the incidence of recurrence. A study was carried out to observe the efficacy of intravesical adjuvant therapy with single immediate versus delayed multi-dose regimen of Mitomycin C (MMC) in preventing recurrence of superficial bladder cancer. One hundred Patients having intermediate risk superficial bladder cancer were randomized into two equal groups. All patients were followed carefully. Total duration of follow-up was minimum 12 months, maximum 36 months, mean 29 months. No recurrence was seen on 3(rd), 6(th) and 9(th) month of intravesical therapy. As much as 94% and 96% of recurrence free rate was observed in immediate single and delayed multi-dose group respectively on 12(th) month; 86% and 84% on 18(th) month; 74% and 72% on 24(th) month; 70% and 68% on 36(th) month of follow-up cystoscopy respectively. Efficacy of post transurethral resection of bladder tumour (TURBT) MMC single immediate dose was found similar to that of MMC delayed multi-dose regimen in preventing the recurrence of intermediate risk superficial bladder transitional cell carcinoma (TCC) in the study. The difference between the two groups insignificant (p>0.05). PMID- 16467762 TI - Effects of stresses on serum cortisol level in Bangladeshi people. AB - The present study was undertaken to estimate serum cortisol level in control experimental groups in Bangladeshi people. For this purpose a total number of 55 subjects of age range (18-55) years were selected from Mymensingh Medical College Hospital during July 2003 to June 2004. Subjects included in this study were divided in to two main groups: control (n=13) and experimental (n=42). According to different types of stress the experimental group was further divided in to four subgroups: infection, psychiatric, pre and post surgery. Blood samples were collected from all subjects with aseptic precautions at 8 AM and serum cortisol concentration was measured by Radioimmunoassay method. Statistical analyses were done by using Student's 't' test. A distinct and statistically significant increased serum cortisol level was observed in infection, psychiatric disorders and post surgical group in comparison to control group (P< 0.001). While no significant difference was observed in pre surgery when compared to that of control group. PMID- 16467763 TI - Electrophoretic patterns of human rotavirus strains prevailing among hospitalized children with acute gastroenteritis. AB - Polyacrylamide gel electrophoresis (PAGE) and silver staining was applied to detect rotavirus dsRNA from acute diarrheic stool of 365 hospitalized children below 5 years of age. The study was conducted at Mymensingh and Sunamgonj districts, Bangladesh from January 2002 to February 2003. Among 345 stool specimens tested, 86 (24.9%) were positive by PAGE. The rate of infection was highest in 0-12 months of age and declined significantly with increasing age. Males were slightly more infected than females and infection rate was more in winter. Twelve different electropherotypes were identified, of them eight were long and four were short. RNA profiles of the analyzed specimens, 88.6% were long and 11.4% were short patterns. Two of these long patterns (2F N 1 L, 2F N 3(U) L) circulated through out the study period and a single type was predominant (2F N 1 L). Mixed electropherotypes were also detected. Electropherotyping technique can be applied routinely to study the prevalence and epidemiological features of rotavirus infection. It is an excellent method for studying genomic variation, tracing mixed infections, detecting atypical rotaviruses lacking group-antigen and characterizing virus strains in outbreaks. PMID- 16467764 TI - Vaccination status of tribal mothers and their under five children. AB - A study was done to find out the vaccination status of the tribal mothers and their under 5 children in some selected villages of Durgapur upazila under Netrakona district. It was a cross sectional study in which 92 tribal mothers and 91 under 5 children were included. The study was carried out in 4 different tribal villages under Netrakona district from February to June 2001. According to National EPI schedule, it was revealed that 58.2% of the children were fully vaccinated, 26.4% incompletely and 15.4% not vaccinated. The individual vaccine coverage was 84.6% for BCG, 68.1% for OPV and DPT, 58.2% for Measles. Considering the literacy, most of the respondents (78.3%) were illiterate and 21.7% had some basic education. None of the mother completed 5 doses of TT coverage. The individual TT coverage was found 78.3% for TT(1), 67.4% for TT(2), 17.4% for TT(3) and 1.1% for TT(4). This study observed that the vaccination status in the tribal children was satisfactory in relation to National coverage, but the vaccination status of the tribal mothers was not satisfactory in our national context. PMID- 16467765 TI - Do all regularly menstruating women ovulate in each cycle? AB - Women may experience anovular menstruation due to some pathophysiological causes which can be detected either by invasive histological examination as well as noninvasive serial ultrasound test. The women who are regularly menstruating without ovulation in each cycle were identified in this study. In a tertiary level infertility care centre of Bangladesh in Dhaka infertile population was the subject of the study. The serial ultrasound noninvasive procedure is used for diagnosis of anovular menstruation and found very much helpful. PMID- 16467766 TI - Long term outcome of birth asphyxiated infants. AB - Fifty eight neonates who survived birth asphyxia were prospectively studied in the department of Paediatrics, Dhaka Medical College Hospital, to see the incidence of long term neurodevelopmental sequelae among them and also to identify possible risk factors. All of them had been admitted in the Neonatal Unit of this hospital and were born between July 1997 & December 1999. After discharge from the hospital, the study infants were followed up at the Child Neurology Clinic of this hospital. They were seen at 6 weeks. at 3 months & then every 3 months upto 2 years of age for evaluation of growth and development (age corrected for prematurity for preterm babies) 69% of them were boys and 31% were girls. 74% of the babies were inborn in the same hospital & the rest were out born. Prolonged labour (24%) and PET (18.96%) were the most common obstetric factors & respiratory distress (38%) and neonatal convulsion (35%) were the important neonatal complications. Of the 58 enlisted study cases, 30 (51.7%) responded to complete 2 years follow up and the rest were lost. Normal growth and developments were observed in 12 (40%) cases, 7 (23.33%) had mild handicap, 6 (20%) moderate & 5 (16.6%) were left with severe neurodevelopmental handicaps at 2 years of age. 61% of the handicapped children had suffered from neonatal seizure for varying periods. It may be concluded that the response to follow up unsatisfactory, the number of severe handicap was alarmingly high and post asphyxiated seizure was associated with poor outcome. So prevention of perinatal asphyxia should be targeted. Further long term follow up study is required. PMID- 16467767 TI - T-cell sensitisation to hepatitis B virus surface antigens. AB - The present study was conducted to explore the immunity to hepatitis B surface antigens (HBsAg) of ad and ay subtypes at the cellular level among adult individuals. Peripheral blood mononuclear cells (PBMCs) obtained from acute hepatitis B virus (HBV) infected patients, chronic HBV infected patients, recovered subjects from HBV infection and uninfected vaccinated controls were stimulated with HBsAg ad and HBsAg ay subtypes in vitro. Stimulated PBMCs were incubated in CO(2) for production of interferon-gamma (IFN-gamma), which was measured from the supernatant of cultured PBMCs by an in-house ELISA technique. The mean +/- SE of IFN-gamma levels produced by PBMCs in response to HBsAg ad among the acute, chronic, recovered and control groups were 282.5+/-134.51 pg/ml, 307.45+/-94.84 pg/ml, 915.62+/-170.80 pg/ml and 511.67+/-161.22 pg/ml respectively, while on stimulation by HBsAg ay, the levels were 246.25+/-103.50 pg/ml, 374.70+/-104.02 pg/ml, 1040 +/-140.76 pg/ml and 465.83+/-166.26 pg/ml respectively among the above mentioned groups. The results of this study showed that PBMCs were non-responsive to stimulation by both HBsAg ad and HBsAg ay subtypes in acute and chronic patients with HBV infection. The recovered group responded significantly to both subtypes of HBsAg and the control group did not. The study indicates that although the patients with acute and chronic HBV infection showed weak or no IFN-gamma response to the HBsAg, subjects showed strong IFN-gamma response to the surface antigens on recovery from HBV infection. PMID- 16467769 TI - A survey of antimicrobial prescribing and dispensing practices in rural Bangladesh. AB - This study was undertaken to determine the patterns of antimicrobial prescription by 64 Rural Medical Practitioners (RMPs) from Bangladesh. The antimicrobial dispensing procedures followed by the local retail drug sellers along with the purchasing capacities of the patients was also assessed. All antimicrobial agents were prescribed mainly on the patient's complaints, and all available antibiotics were prescribed in inappropriate doses and duration. In most cases, the RMPs initiated treatment with a parenteral form of antibiotic, and a different oral antibiotic usually followed. Parenteral streptomycin was used most frequently in short inadequate courses. Almost half of the antibiotics were sold without any prescriptions, and even ordinary people without any knowledge of medicine asked the drug seller for specific antibiotics. This unregulated prescribing and dispensing practice has the potential risk for the development and spread of antimicrobial resistance on a global scale. PMID- 16467768 TI - Study of oral itraconazole and terbinafine pulse therapy in onychomycosis. AB - In this open, randomized and comparative study, the safety and efficacy of systemic intermittent itraconazole and terbinafine was examined in 30 patients with onychomycosis. The patient with positive mycological culture and also the patients with positive microscopy and negative culture were investigated. Patients were randomly assigned: 15 patients in each group received either 200mg itraconazole or 250 mg terbinafine twice daily during the first week of a 4 weeks cycle. The treatment duration was 16 weeks and was followed-up for 36 weeks. Both the treatment regimen showed significant reduction in onychomycosis affected areas after 8 weeks and maximum reduction was observed at the end of 36 weeks. At the end point of the follow-up period, the clinical cure rates (no residual deformity or with some deformity) were 86.7% in the itraconazole group and 100% in the terbinafine group. The mycological cure rates were 86.7% and 100% respectively. However, no statistically significant differences between the treatment groups were seen in clinical, mycological (P= 0.864) and severity assessment (P= 0.220). Nausea, abdominal cramp, headache, back pain and flu like syndrome are the adverse effects more frequently reported. At least one adverse effect was reported by 17 patients, of them 12 belonged to itraconazole group and 5 to terbinafine group and the difference was statistically significant (P= 0.027). The overall therapeutic effectiveness, safety and cost affectivity were in favor of Terbinafine pulse therapy. PMID- 16467770 TI - Effectiveness of montelukast in the treatment of atopic dermatitis. AB - Leukotriene receptor antagonist ( montelukast) are recommended for the treatment of asthma, and have proved anecdotally successfully even in atopic dermatitis. In this open randomized clinical trial, the efficacy and safety of montelukast were assessed in the atopic dermatitis. Out of 31 enrolled patients all completed the study among which 16 in the montelukast group and 15 in the control group. No patient dropped from the study. Statistically significant SCORAD improvement (P = 0.003) was observed in montelukast group but in the control group SCORAD improvement was not statistically significant (P = 0.088). According to the patients impression pruritus was the most influenced SCORAD item by montelukast group immediately followed by sleep loss and inflammatory signs. On the contrary montelukast seemed to be completely devoid of activity on xerosis. No adverse effect of montelukast was observed in the present study. PMID- 16467771 TI - Repair of cleft lip on relatively older children with anthropometrical surgical outcome. AB - A prospective study was done on 54 patients aged over 6 months with cleft lip with or without cleft palate admitted in the department of pediatric surgery of Mymensingh Medical College Hospital during the period from Oct 2002 to Dec 2004. The anthropometrical surgical outcome was evaluated by measuring the length, diameter and thickness of hemilip and diameter of nostril both pre and postoperatively (according to schedule). Patients were categorized into three groups according to their age. Mean Corrective rate was evaluated and it was shown 95% in the 1(st) age group, 93% in 2(nd) age group and 95.55% in the 3(rd) group. Complication was evaluated in only 16.66% of patients. Regarding the out come scar, cupid's bow, notching, labial and nasal symmetry were taken in consideration. Beside these this study might play a comparative role with the results of correction of cleft lip in early age. PMID- 16467772 TI - A case report of drinking glass in rectum. AB - A fifty years old man was admitted in the Dept. of Surgery of Community Based Medical College, Bangladesh, Hospital with the history of pain and distension of lower abdomen for 4 days with obstipation for the same duration. He gave history of being assaulted and forceful introduction of some foreign body per-rectally 5 days back. History, clinical examination and abdominal X-rays diagnosed the case as impacted foreign body in rectum. He was undergone laparotomy and a big drinking glass was extracted from sigmoid colon. Patient developed subcutaneous stitch infection postoperatively and discharged from the hospital after controlling the infection. PMID- 16467773 TI - Retrograde jejunogastric intussusception (JGI) with strangulation following previous gastrojejunostomy. AB - Jejunogastric intussusception is a rare but potentially very serious complication of gastrectomy or gastrojejunostomy. To avoid mortality early diagnosis and prompt surgical intervention is mandatory. A young man presented with epigastric pain and bilous vomiting followed by haematemesis 15 years after vagotomy & gastrojejunostomy for chronic duodenal ulcer. At presentation the patient was in shock and an emergency laparotomy was done after resuscitation. At laparotomy a retrograde type II JGI was found and managed by resection of the affected segment and partial gastrectomy and jejuno - jejunostomy with closure of the duodenal stump. Postoperative recovery was uneventful. Retrograde JGI is a rare condition and only less than 200 cases have been reported since its first report. Clinical picture of acute intestinal obstruction with suspicion about the condition in patients having a past history of gastrojejunostomy makes the elusive diagnosis definite and demands early surgery to reduce the grave consequences of the disease. PMID- 16467774 TI - Combined occurrence of hyperthyroid Graves' and Hashimoto's thyroiditis. AB - A 40 years old, married Govt. servant from Sadar upazila, Mymensingh was admitted in Mymensingh Medical College Hospital on 9(th) February, 2005 with the complaints of excessive sweating for 1 year, gradual loss of weight for 6 months, swelling in front of the neck for 1(1/2) months, and hoarseness of voice for 1 month. He was nervous, irritable, emotionally labile. Thyroid gland was symmetrically enlarged, firm in consistency with scalloped surface. Palms were warm and sweaty with fine tremor in outstretched hands. Lid lag, lid retraction and proptosis were the occular manifestations. All the reflexes were exaggerated. Radioactive iodine uptake showed enlarged gland with homogenously increased radiotracer concentration, ultrasonogram findings were enlarged gland with hypoechoic parenchyma with fibrous septa, T(3), T(4), TSH values were 6.56 nmol/L, 241.09 nmol/L and 0.14 mIU/L respectively. Thyroid microsomal antibody level was 32.87%. Thyroid FNAC findings were sheets of regular follicular cells, some large cells with granular basophilic cytoplasm, macrophages, a few inflammatory cells and giant cells. All the above findings were in favour of a diagnosis of hyperthyroid Graves' with Hashimoto's thyroiditis. PMID- 16467775 TI - Gnathostomiasis: a rare nematode infection. AB - Gnathostoma, primarily an animal nematode, can infect human by the third stage larva by consumption of undercooked or raw fish, poultry, or pork. In Bangladesh gnathostomiasis is a very rare condition. The first case, an ocular infection by gnathostoma was reported in 2001 from Rangpur, a northern district. This is the second case report of gnathostomiasis which also is an ocular infection, occurred in an area of greater district of Rangpur. A female patient of 32 year of age of the northern district, Nilphamari has got infected with a species of Gnathostoma, manifested by the appearance of a live larva near the right lateral margin of anterior surface of iris of her right eye. The larva was removed surgically from her eye and the patient was cured from symptoms. PMID- 16467776 TI - Neonatal sepsis-- a global problem: an overview. AB - Neonatal sepsis is one of the major health problems throughout the world. Every year an estimated 30 million newborns acquire infection and 1-2 million of these die. The present review provides updates regarding neonatal sepsis to help paediatricians to protect the newborn from this deadly problem. The onset of sepsis within first 48 hours of life (early onset sepsis) is frequently associated with pre and perinatal predisposing factors while onset after 48-72 hours of life (late onset sepsis) frequently reflects infection acquired nosocomially. Some literatures say that early onset disease presents in the first 5-7 days of life. Klebsiella pneumoniae is the leading pathogen causing neonatal sepsis in Bangladesh and neighbouring countries. Among many risk factors the single most important neonatal risk factor is low birth weight. Other main risk factors are invassive procedures in the postnatal period and inadequate hand washing before and after handling babies. Sepsis score is a useful method for early and rapid diagnosis of neonatal sepsis which was developed by Tollner U in 1982. Antibiotics should be given to most of the neonates suspected of infection. Ampicillin and gentamicin are the first drug of choice. In Bangladesh context sepsis score may be used as a good parameter for the early and rapid diagnosis of sepsis and that will guide the treatment plan. Clean and safe delivery, early and exclusive breastfeeding, strict postnatal cleanliness following adequate handwashing and aseptic technique during invasive procedure might reduce the incidence of neonatal sepsis. Prompt use of antibiotic according to standard policy is warranted to save the newborn lives from septicaemia. PMID- 16467777 TI - Racial/ethnic disparities in diagnoses of HIV/AIDS--33 states, 2001-2004. AB - In 2003, an estimated 1.2 million persons in the United States were living with human immunodeficiency virus (HIV) infection, 47% of whom were non-Hispanic blacks. This report describes racial/ethnic disparities in diagnoses of HIV/acquired immunodeficiency syndrome (AIDS) during 2001-2004 and reported to CDC through June 2005 by 33 states that used confidential, name-based reporting of HIV and AIDS cases for at least 4 years. Of the estimated 157,252 diagnoses of HIV infection, the number of cases and diagnosis rates among blacks were higher than those for all other racial/ethnic populations combined. Among males, blacks had the largest or second-largest percentage of cases in every transmission category; among females, blacks had the largest percentage of cases in all transmission categories. Moreover, among both males and females, blacks represented the largest percentage of HIV/AIDS diagnoses in every age group. New and improved prevention strategies, including expanded HIV testing, targeted communications, and tailored prevention services, are needed to help address disparities in HIV transmission among blacks. PMID- 16467778 TI - Brief report: Leptospirosis after flooding of a university campus--Hawaii, 2004. AB - On November 19, 2004, the Hawaii Department of Health (HDOH) received a report that a University of Hawaii professor aged 56 years had been hospitalized with suspected leptospirosis after cleaning his flooded laboratory. On October 31, heavy rains had caused an adjacent stream to overflow its banks and flood the campus. Persons exposed to fresh water or mud contaminated by the urine of animals infected with the spirochete Leptospira interrogans can have systemic illness if the leptospires enter the body through broken skin or mucous membranes. This report describes the subsequent investigation by HDOH, assisted by CDC, which highlights the importance of maintaining clinical suspicion for leptospirosis after flooding in areas where the illness is endemic, even in well developed urban settings. PMID- 16467779 TI - New laboratory assay for diagnostic testing of avian influenza A/H5 (Asian Lineage). AB - On February 3, 2006, the Food and Drug Administration (FDA) announced clearance of the Influenza A/H5 (Asian Lineage) Virus Real-Time Reverse Transcription Polymerase Chain Reaction (RT-PCR) Primer and Probe Set and inactivated virus as a source of positive RNA control for the in vitro qualitative detection of highly pathogenic influenza A/H5 virus (Asian lineage). Two genetic lineages of influenza A/H5 viruses exist: Eurasian (Asian) and North American. The primer and probe set, developed at CDC, is designed to detect highly pathogenic influenza A/H5 viruses from the Asian lineage associated with recent laboratory-confirmed infections of avian influenza in humans in east Asia and, most recently, in Turkey and Iraq. PMID- 16467780 TI - Update: Influenza activity--United States, January 22-28, 2006. AB - During January 22-28, 2006, the number of states reporting widespread influenza activity remained at five. Twenty-one states reported regional activity, 13 reported local activity, and 11 reported sporadic activity. PMID- 16467781 TI - Protection of adult mouse progenitor cells and human glioma cells by de novo decorin expression in an oxygen- and glucose-deprived cell culture model system. AB - We employed an in vitro hypoxia cell culture model system and gene transfer technology to examine the effect of the decorin gene on cell survival against oxygen and glucose deprivation (OGD). Ectopic expression of decorin in subventricular zone (SVZ) cells from adult male mouse brain and human glioblastoma U-87 cells kept the cells viable against 24 h of OGD. Fewer than 1% of decorin-synthesizing cells were apoptotic after 12 h of OGD. In contrast, 100% of the control cells were apoptotic even after 4 h of OGD. De novo decorin synthesis in SVZ and U-87 cells induced expression of p21, p27 and Ras, AKT (acutely transforming retrovirus AKT8 in rodent T-cell lymphoma), and phosphorylated AKT. Blocking of phosphoinositide 3-kinase (PI-3K), Ras, and the epidermal growth factor receptor with specific inhibitors had no effect on induction of Ras, p21, and p27 at the messenger RNA level in decorin-synthesizing SVZ and U-87 cells. PI-3K inhibitors significantly increased apoptosis in decorin expressing cells. Our data indicate that induction of p21, p27, Ras, AKT, and phosphorylated AKT by decorin inhibits apoptosis and protects U-87 and SVZ cells against OGD. Therefore, our data suggest that decorin is a potent trophic factor that protects neuronal progenitor cells and glioma cells from OGD. PMID- 16467782 TI - The role of endothelial nitric oxide synthase (eNOS) genetic variants in European patients with intracranial aneurysms. AB - The endothelial nitric oxide synthase (eNOS) gene harbors three well characterized genetic variants, which have been reported to be associated with various vascular diseases. Recently, conflicting results have been published relating to the role of these genetic variants in the pathogenesis of intracranial aneurysms (IA). Therefore, we analyzed these variants in a large European population of IA patients and controls. In all, 142 patients with IA and 190 controls were enrolled in our study. The -786T>C and 894G>T single-nucleotide polymorphisms (SNPs) were analyzed by direct sequencing of the corresponding sections in the genomic DNA. A variable number tandem repeat (VNTR) located in intron 4 of the gene and consisting of either four or five 27-base pair (bp) repeats was analyzed by polymerase chain reaction amplification and electrophoresis using ALF sequencertrade mark equipment. Genotype and allele frequencies were determined, and the frequencies in cases and controls were compared. In addition, haplotypes were constructed. There were no deviations from Hardy-Weinberg equilibrium. Genotype and allele frequencies did not differ significantly between cases and controls in any sample group or after stratification for multiple IA or aneurysm size. No single haplotype was significantly associated with the phenotype of an IA. The -786T>C, 894G>T, and 27 bp VNTR genetic variants of the eNOS gene are not associated with IA in the European population. PMID- 16467783 TI - Glucose is necessary to maintain neurotransmitter homeostasis during synaptic activity in cultured glutamatergic neurons. AB - Glucose is the primary energy substrate for the adult mammalian brain. However, lactate produced within the brain might be able to serve this purpose in neurons. In the present study, the relative significance of glucose and lactate as substrates to maintain neurotransmitter homeostasis was investigated. Cultured cerebellar (primarily glutamatergic) neurons were superfused in medium containing [U-13C]glucose (2.5 mmol/L) and lactate (1 or 5 mmol/L) or glucose (2.5 mmol/L) and [U-13C]lactate (1 mmol/L), and exposed to pulses of N-methyl-D-aspartate (300 micromol/L), leading to synaptic activity including vesicular release. The incorporation of 13C label into intracellular lactate, alanine, succinate, glutamate, and aspartate was determined by mass spectrometry. The metabolism of [U-13C]lactate under non-depolarizing conditions was high compared with that of [U-13C]glucose; however, it decreased significantly during induced depolarization. In contrast, at both concentrations of extracellular lactate, the metabolism of [U-13C]glucose was increased during neuronal depolarization. The role of glucose and lactate as energy substrates during vesicular release as well as transporter-mediated influx and efflux of glutamate was examined using preloaded D-[3H]aspartate as a glutamate tracer and DL-threo-beta benzyloxyaspartate to inhibit glutamate transporters. The results suggest that glucose is essential to prevent depolarization-induced reversal of the transporter (efflux), whereas vesicular release was unaffected by the choice of substrate. In conclusion, the present study shows that glucose is a necessary substrate to maintain neurotransmitter homeostasis during synaptic activity and that synaptic activity does not induce an upregulation of lactate metabolism in glutamatergic neurons. PMID- 16467784 TI - Bilirubin oxidation products (BOXes) and their role in cerebral vasospasm after subarachnoid hemorrhage. AB - Many factors have been postulated to cause delayed subarachnoid hemorrhage (SAH) induced vasospasm, including hemoglobin, nitric oxide, endothelin, and free radicals. We propose that free radicals (because of the high levels that are produced in the blood clots surrounding blood vessels after SAH) act on bilirubin, biliverdin, and possibly heme to produce BOXes (Bilirubin OXidized Products). Bilirubin oxidation products act on vascular smooth muscle cells to produce chronic vasoconstriction and vasospasm combined with a vasculopathy because of smooth muscle cell injury. This review summarizes recent evidence that BOXes play a role in SAH-induced vasospasm. The data supporting a role for BOXes includes (1) identification of molecules in cerebrospinal fluid (CSF) of patients with vasospasm after SAH that have structures consistent with BOXes; (2) BOXes are vasoactive in vitro and mimic the biochemical actions of CSF of patients with vasospasm; (3) BOXes are vasoactive in vivo, constricting rat cerebral vessels; and (4) there is a correlation between clinical occurrence of vasospasm and BOXes concentration in our preliminary study of patients with SAH. Since oxidation of bilirubin, biliverdin, and perhaps heme is proposed to produce BOXes that contribute to vasospasm, either blocking bilirubin formation, inactivating bilirubin or BOXes, or removing all of the blood clot before vasospasm are potential treatment targets. PMID- 16467785 TI - Prognostic importance of ambulatory blood pressure recordings in patients with chronic kidney disease. AB - Ambulatory systolic blood pressure (BP) correlates better with risk factors for progression of chronic kidney disease (CKD) compared to clinic measured BP, but its role in predicting end-stage renal disease (ESRD) and death in patients with CKD is unknown. In a cohort study of 217 Veterans with CKD BP was measured by ambulatory monitoring and in the clinic. Twenty-four hour ambulatory BP was 133.5 +/- 16.6/73.1 +/- 11.1 mm Hg and clinic BP was 155.2 +/- 25.6/84.7 +/- 14.2 mm Hg. The composite renal end point of ESRD or death over a median follow-up of 3.5 years occurred in 75 patients (34.5%), death occurred in 52 patients (24.0%), and ESRD in 36/178 patients (20.2%). Thirty-nine patients died before reaching ESRD. One standard deviation (s.d.) increase in systolic BP increased the risk of composite outcome to 1.69 (95% confidence interval (CI) 1.32-2.17) for standard clinic measurement and to 1.88 (95% CI 1.48-2.39) for 24 h ambulatory BP recording. One s.d. increase in 24 h ambulatory systolic BP increased the risk of ESRD to 3.04 (95% CI 2.13-4.35) and to 2.20 (95% CI 1.43-3.39) when adjusted for standard clinic systolic BP. Non-dipping was associated with increased risk of total mortality and composite end point. In patients with CKD, BPs obtained by ambulatory monitoring are a stronger predictor of ESRD or death compared to BPs obtained in the clinic. Systolic ambulatory BP and nondipping are independent predictors for ESRD after adjusting for clinic BP. However, adjustment for other risk factors for CKD progression removes the independent prognostic value of ambulatory BP. PMID- 16467786 TI - NK cells do not mediate renal injury in murine adriamycin nephropathy. AB - In adriamycin nephropathy (AN), a model of chronic proteinuric renal injury, the absence of functional B and T cells with residual natural killer (NK) cells, and macrophages in severe combined immunodeficient (SCID) mice results in more severe disease than in immunocompetent mice. We have recently shown expression of the stimulatory NK cell molecule NKG2D and its ligand RAE-1 in the adriamycin (ADR) kidney. Therefore, we sought to determine the role of NK cells in AN. We used anti-asialo GM1 NK cell depletion in immunocompetent BALB/c mice with AN, and also compared AN in immunodeficient SCID mice and immunodeficient nonobese diabetic (NOD)-SCID mice (that have impaired NK cell function). The number of NK cells was increased in AN in BALB/c mice compared with normal controls. NK cell depletion or reduction of NK function in NOD-SCID mice did not affect the severity of disease. In both wild type and immunodeficient models, ADR upregulated RAE-1 in the kidney. High levels of Class I major histocompatibility complex molecules were found in both models of AN. In conclusion, NK cells do not play a significant role in AN. PMID- 16467787 TI - Enterobacteriaceae peritonitis complicating peritoneal dialysis: a review of 210 consecutive cases. AB - Enterobacteriaceae peritonitis is a serious complication in peritoneal dialysis (PD), but the clinical course of PD-related Enterobacteriaceae peritonitis remains unclear. We reviewed all Enterobacteriaceae peritonitis in our dialysis unit from 1995 to 2004. During this period, there were 1748 episodes of peritonitis recorded; 210 episodes (12.0%) in 123 patients were caused by Enterobacteriaceae. The most common species was Escherichia coli, accounting for 111 episodes. The primary response rate was 84.8% and complete cure rate was 58.1%. The presence of exit site infection was associated with a lower complete cure rate (43.2 versus 61.3%, P = 0.034). A total of 82 episodes (39.0%) did not respond to single antibiotic treatment despite sensitivity in vitro, and a second antibiotic was added. Patients treated with two antibiotics had a marginally lower risk of relapse and recurrence than those with one antibiotic (21.4 versus 36.1%, P = 0.051). The episodes that had recent antibiotic therapy had a marginally lower complete cure rate (49.3 versus 62.8%, P = 0.06). There was a gradual increase in the prevalence of resistance to several commonly used antibiotics over the years. Recent antibiotic therapy was associated with resistance to cefotaxime, ceftazidime, cefoperazone/sulbactam, and piperacillin/tazobactam. We conclude that Enterobacteriaceae peritonitis is a serious complication of PD. Recent antibiotic therapy is the major risk factor of antibiotic resistance. Exit site infection, and probably recent antibiotic therapy, is associated with poor therapeutic response. Contrary to the current recommendation, treatment with two antibiotics may reduce the risk of relapse and recurrence. PMID- 16467789 TI - Atomic structure of a Na+- and K+-conducting channel. AB - Ion selectivity is one of the basic properties that define an ion channel. Most tetrameric cation channels, which include the K+, Ca2+, Na+ and cyclic nucleotide gated channels, probably share a similar overall architecture in their ion conduction pore, but the structural details that determine ion selection are different. Although K+ channel selectivity has been well studied from a structural perspective, little is known about the structure of other cation channels. Here we present crystal structures of the NaK channel from Bacillus cereus, a non-selective tetrameric cation channel, in its Na+- and K+-bound states at 2.4 A and 2.8 A resolution, respectively. The NaK channel shares high sequence homology and a similar overall structure with the bacterial KcsA K+ channel, but its selectivity filter adopts a different architecture. Unlike a K+ channel selectivity filter, which contains four equivalent K+-binding sites, the selectivity filter of the NaK channel preserves the two cation-binding sites equivalent to sites 3 and 4 of a K+ channel, whereas the region corresponding to sites 1 and 2 of a K+ channel becomes a vestibule in which ions can diffuse but not bind specifically. Functional analysis using an 86Rb flux assay shows that the NaK channel can conduct both Na+ and K+ ions. We conclude that the sequence of the NaK selectivity filter resembles that of a cyclic nucleotide-gated channel and its structure may represent that of a cyclic nucleotide-gated channel pore. PMID- 16467794 TI - Physics fights back. PMID- 16467793 TI - Network of concern. PMID- 16467795 TI - Japan's research conduct. PMID- 16467797 TI - Drop in HIV infection rate used to justify focus on morality. PMID- 16467798 TI - Dealer unearths Hooke's Royal Society notes. PMID- 16467799 TI - Possible planets left with no name. PMID- 16467800 TI - Political strife set to delay EU funds. PMID- 16467801 TI - One place, one parent, two species. PMID- 16467804 TI - Space rocks wanted: cash paid. PMID- 16467803 TI - Journal lays bare remarks from peer reviewers. PMID- 16467806 TI - Physics wins the US budget race. PMID- 16467807 TI - The scientific balance of power. PMID- 16467788 TI - Sympatric speciation in palms on an oceanic island. AB - The origin of species diversity has challenged biologists for over two centuries. Allopatric speciation, the divergence of species resulting from geographical isolation, is well documented. However, sympatric speciation, divergence without geographical isolation, is highly controversial. Claims of sympatric speciation must demonstrate species sympatry, sister relationships, reproductive isolation, and that an earlier allopatric phase is highly unlikely. Here we provide clear support for sympatric speciation in a case study of two species of palm (Arecaceae) on an oceanic island. A large dated phylogenetic tree shows that the two species of Howea, endemic to the remote Lord Howe Island, are sister taxa and diverged from each other well after the island was formed 6.9 million years ago. During fieldwork, we found a substantial disjunction in flowering time that is correlated with soil preference. In addition, a genome scan indicates that few genetic loci are more divergent between the two species than expected under neutrality, a finding consistent with models of sympatric speciation involving disruptive/divergent selection. This case study of sympatric speciation in plants provides an opportunity for refining theoretical models on the origin of species, and new impetus for exploring putative plant and animal examples on oceanic islands. PMID- 16467808 TI - Fractals and art: in the hands of a master. PMID- 16467810 TI - Cloning: do we even need eggs? PMID- 16467809 TI - Cloning: mining the secrets of the egg. PMID- 16467811 TI - No end in sight for stem-cell odyssey. PMID- 16467814 TI - Problems at plutonium lab need more than a quick fix. PMID- 16467815 TI - Dwindling fish numbers already of concern in 1883. PMID- 16467816 TI - No unfairness in funding of Croatian minister's project. PMID- 16467817 TI - Climate may not be linked with circulation slowdown. PMID- 16467822 TI - Palaeontology: a Jurassic tyrant is crowned. PMID- 16467823 TI - Chemical biology: aptamers in nanoland. PMID- 16467825 TI - Immunology: exhausted T cells perk up. PMID- 16467826 TI - Earth science: the rise and growth of Tibet. PMID- 16467827 TI - Semiconductor physics: transport news. PMID- 16467828 TI - Evolution: memories of mammoths. PMID- 16467829 TI - Volcanoes and climate: Krakatoa's signature persists in the ocean. AB - We have analysed a suite of 12 state-of-the-art climate models and show that ocean warming and sea-level rise in the twentieth century were substantially reduced by the colossal eruption in 1883 of the volcano Krakatoa in the Sunda strait, Indonesia. Volcanically induced cooling of the ocean surface penetrated into deeper layers, where it persisted for decades after the event. This remarkable effect on oceanic thermal structure is longer lasting than has previously been suspected and is sufficient to offset a large fraction of ocean warming and sea-level rise caused by anthropogenic influences. PMID- 16467830 TI - Palaeo-altimetry of the late Eocene to Miocene Lunpola basin, central Tibet. AB - The elevation history of the Tibetan plateau provides direct insight into the tectonic processes associated with continent-continent collisions. Here we present oxygen-isotope-based estimates of the palaeo-altimetry of late Eocene and younger deposits of the Lunpola basin in the centre of the plateau, which indicate that the surface of Tibet has been at an elevation of more than 4 kilometres for at least the past 35 million years. We conclude that crustal, but not mantle, thickening models, combined with plate-kinematic solutions of India Asia convergence, are compatible with palaeo-elevation estimates across the Tibetan plateau. PMID- 16467832 TI - Anti-planetward auroral electron beams at Saturn. AB - Strong discrete aurorae on Earth are excited by electrons, which are accelerated along magnetic field lines towards the planet. Surprisingly, electrons accelerated in the opposite direction have been recently observed. The mechanisms and significance of this anti-earthward acceleration are highly uncertain because only earthward acceleration was traditionally considered, and observations remain limited. It is also unclear whether upward acceleration of the electrons is a necessary part of the auroral process or simply a special feature of Earth's complex space environment. Here we report anti-planetward acceleration of electron beams in Saturn's magnetosphere along field lines that statistically map into regions of aurora. The energy spectrum of these beams is qualitatively similar to the ones observed at Earth, and the energy fluxes in the observed beams are comparable with the energies required to excite Saturn's aurora. These beams, along with the observations at Earth and the barely understood electron beams in Jupiter's magnetosphere, demonstrate that anti-planetward acceleration is a universal feature of aurorae. The energy contained in the beams shows that upward acceleration is an essential part of the overall auroral process. PMID- 16467831 TI - Discovery of very-high-energy gamma-rays from the Galactic Centre ridge. AB - The source of Galactic cosmic rays (with energies up to 10(15) eV) remains unclear, although it is widely believed that they originate in the shock waves of expanding supernova remnants. At present the best way to investigate their acceleration and propagation is by observing the gamma-rays produced when cosmic rays interact with interstellar gas. Here we report observations of an extended region of very-high-energy (> 10(11) eV) gamma-ray emission correlated spatially with a complex of giant molecular clouds in the central 200 parsecs of the Milky Way. The hardness of the gamma-ray spectrum and the conditions in those molecular clouds indicate that the cosmic rays giving rise to the gamma-rays are likely to be protons and nuclei rather than electrons. The energy associated with the cosmic rays could have come from a single supernova explosion around 10(4) years ago. PMID- 16467833 TI - Electronic transport in nanometre-scale silicon-on-insulator membranes. AB - The widely used 'silicon-on-insulator' (SOI) system consists of a layer of single crystalline silicon supported on a silicon dioxide substrate. When this silicon layer (the template layer) is very thin, the assumption that an effectively infinite number of atoms contributes to its physical properties no longer applies, and new electronic, mechanical and thermodynamic phenomena arise, distinct from those of bulk silicon. The development of unusual electronic properties with decreasing layer thickness is particularly important for silicon microelectronic devices, in which (001)-oriented SOI is often used. Here we show- using scanning tunnelling microscopy, electronic transport measurements, and theory--that electronic conduction in thin SOI(001) is determined not by bulk dopants but by the interaction of surface or interface electronic energy levels with the 'bulk' band structure of the thin silicon template layer. This interaction enables high-mobility carrier conduction in nanometre-scale SOI; conduction in even the thinnest membranes or layers of Si(001) is therefore possible, independent of any considerations of bulk doping, provided that the proper surface or interface states are available to enable the thermal excitation of 'bulk' carriers in the silicon layer. PMID- 16467834 TI - Initial corrosion observed on the atomic scale. AB - Corrosion destroys more than three per cent of the world's GDP. Recently, the electrochemical decomposition of metal alloys has been more productively harnessed to produce porous materials with diverse technological potential. High resolution insight into structure formation during electrocorrosion is a prerequisite for an atomistic understanding and control of such electrochemical surface processes. Here we report atomic-scale observations of the initial stages of corrosion of a Cu3Au111 single crystal alloy within a sulphuric acid solution. We monitor, by in situ X-ray diffraction with picometre-scale resolution, the structure and chemical composition of the electrolyte/alloy interface as the material decomposes. We reveal the microscopic structural changes associated with a general passivation phenomenon of which the origin has been hitherto unclear. We observe the formation of a gold-enriched single-crystal layer that is two to three monolayers thick, and has an unexpected inverted (CBA-) stacking sequence. At higher potentials, we find that this protective passivation layer dewets and pure gold islands are formed; such structures form the templates for the growth of nanoporous metals. Our experiments are carried out on a model single-crystal system. However, the insights should equally apply within a crystalline grain of an associated polycrystalline electrode fabricated from many other alloys exhibiting a large difference in the standard potential of their constituents, such as stainless steel (see ref. 5 for example) or alloys used for marine applications, such as CuZn or CuAl. PMID- 16467835 TI - Winter forest soil respiration controlled by climate and microbial community composition. AB - Most terrestrial carbon sequestration at mid-latitudes in the Northern Hemisphere occurs in seasonal, montane forest ecosystems. Winter respiratory carbon dioxide losses from these ecosystems are high, and over half of the carbon assimilated by photosynthesis in the summer can be lost the following winter. The amount of winter carbon dioxide loss is potentially susceptible to changes in the depth of the snowpack; a shallower snowpack has less insulation potential, causing colder soil temperatures and potentially lower soil respiration rates. Recent climate analyses have shown widespread declines in the winter snowpack of mountain ecosystems in the western USA and Europe that are coupled to positive temperature anomalies. Here we study the effect of changes in snow cover on soil carbon cycling within the context of natural climate variation. We use a six-year record of net ecosystem carbon dioxide exchange in a subalpine forest to show that years with a reduced winter snowpack are accompanied by significantly lower rates of soil respiration. Furthermore, we show that the cause of the high sensitivity of soil respiration rate to changes in snow depth is a unique soil microbial community that exhibits exponential growth and high rates of substrate utilization at the cold temperatures that exist beneath the snow. Our observations suggest that a warmer climate may change soil carbon sequestration rates in forest ecosystems owing to changes in the depth of the insulating snow cover. PMID- 16467836 TI - A basal tyrannosauroid dinosaur from the Late Jurassic of China. AB - The tyrannosauroid fossil record is mainly restricted to Cretaceous sediments of Laurasia, although some very fragmentary Jurassic specimens have been referred to this group. Here we report a new basal tyrannosauroid, Guanlong wucaii gen. et sp. nov., from the lower Upper Jurassic of the Junggar Basin, northwestern China. G. wucaii is the oldest known tyrannosauroid and shows several unexpectedly primitive pelvic features. Nevertheless, the limbs of G. wucaii share several features with derived coelurosaurs, and it possesses features shared by other coelurosaurian clades. This unusual combination of character states provides an insight into the poorly known early radiation of the Coelurosauria. Notably, the presumed predatory Guanlong has a large, fragile and highly pneumatic cranial crest that is among the most elaborate known in any non-avian dinosaur and could be comparable to some classical exaggerated ornamental traits among vertebrates. PMID- 16467837 TI - Sympatric speciation in Nicaraguan crater lake cichlid fish. AB - Sympatric speciation, the formation of species in the absence of geographical barriers, remains one of the most contentious concepts in evolutionary biology. Although speciation under sympatric conditions seems theoretically possible, empirical studies are scarce and only a few credible examples of sympatric speciation exist. Here we present a convincing case of sympatric speciation in the Midas cichlid species complex (Amphilophus sp.) in a young and small volcanic crater lake in Nicaragua. Our study includes phylogeographic, population-genetic (based on mitochondrial DNA, microsatellites and amplified fragment length polymorphisms), morphometric and ecological analyses. We find, first, that crater Lake Apoyo was seeded only once by the ancestral high-bodied benthic species Amphilophus citrinellus, the most common cichlid species in the area; second, that a new elongated limnetic species (Amphilophus zaliosus) evolved in Lake Apoyo from the ancestral species (A. citrinellus) within less than approximately 10,000 yr; third, that the two species in Lake Apoyo are reproductively isolated; and fourth, that the two species are eco-morphologically distinct. PMID- 16467838 TI - Localized maternal orthodenticle patterns anterior and posterior in the long germ wasp Nasonia. AB - The Bicoid (Bcd) gradient in Drosophila has long been a model for the action of a morphogen in establishing embryonic polarity. However, it is now clear that bcd is a unique feature of higher Diptera. An evolutionarily ancient gene, orthodenticle (otd), has a bcd-like role in the beetle Tribolium. Unlike the Bcd gradient, which arises by diffusion of protein from an anteriorly localized messenger RNA, the Tribolium Otd gradient forms by translational repression of otd mRNA by a posteriorly localized factor. These differences in gradient formation are correlated with differences in modes of embryonic patterning. Drosophila uses long germ embryogenesis, where the embryo derives from the entire anterior-posterior axis, and all segments are patterned at the blastoderm stage, before gastrulation. In contrast, Tribolium undergoes short germ embryogenesis: the embryo arises from cells in the posterior of the egg, and only anterior segments are patterned at the blastoderm stage, with the remaining segments arising after gastrulation from a growth zone. Here we describe the role of otd in the long germband embryo of the wasp Nasonia vitripennis. We show that Nasonia otd maternal mRNA is localized at both poles of the embryo, and resulting protein gradients pattern both poles. Thus, localized Nasonia otd has two major roles that allow long germ development. It activates anterior targets at the anterior of the egg in a manner reminiscent of the Bcd gradient, and it is required for pre-gastrulation expression of posterior gap genes. PMID- 16467839 TI - Whole-cell patch-clamp measurements of spermatozoa reveal an alkaline-activated Ca2+ channel. AB - In mammals, sperm cells become motile during ejaculation and swim up the female reproductive tract. Before fertilization and to overcome various barriers, their motility must be hyperactivated, a motion that is characterized by vigorous asymmetric tail beating. Hyperactivation requires an increase in calcium in the flagella, a process that probably involves plasmalemmal ion channels. Numerous attempts in the past two decades to understand sperm cell channels have been frustrated by the difficulty of measuring spermatozoan transmembrane ion currents. Here, by using a simple approach to patch-clamp spermatozoa and to characterize whole-spermatozoan currents, we describe a constitutively active flagellar calcium channel that is strongly potentiated by intracellular alkalinization. This current is not present in spermatozoa lacking the sperm specific putative ion channel protein, CatSper1. This plasma membrane protein of the six transmembrane-spanning ion channel superfamily is specifically localized to the principal piece of the sperm tail and is required for sperm cell hyperactivation and male fertility. Our results identify CatSper1 as a component of the key flagellar calcium channel, and suggest that intracellular alkalinization potentiates CatSper current to increase intraflagellar calcium and induce sperm hyperactivation. PMID- 16467840 TI - Molecular characterization of Ph1 as a major chromosome pairing locus in polyploid wheat. AB - The foundation of western civilization owes much to the high fertility of bread wheat, which results from the stability of its polyploid genome. Despite possessing multiple sets of related chromosomes, hexaploid (bread) and tetraploid (pasta) wheat both behave as diploids at meiosis. Correct pairing of homologous chromosomes is controlled by the Ph1 locus. In wheat hybrids, Ph1 prevents pairing between related chromosomes. Lack of Ph1 activity in diploid relatives of wheat suggests that Ph1 arose on polyploidization. Absence of phenotypic variation, apart from dosage effects, and the failure of ethylmethane sulphonate treatment to yield mutants, indicates that Ph1 has a complex structure. Here we have localized Ph1 to a 2.5-megabase interstitial region of wheat chromosome 5B containing a structure consisting of a segment of subtelomeric heterochromatin that inserted into a cluster of cdc2-related genes after polyploidization. The correlation of the presence of this structure with Ph1 activity in related species, and the involvement of heterochromatin with Ph1 (ref. 6) and cdc2 genes with meiosis, makes the structure a good candidate for the Ph1 locus. PMID- 16467841 TI - ClpS is an essential component of the N-end rule pathway in Escherichia coli. AB - The N-end rule states that the half-life of a protein is determined by the nature of its amino-terminal residue. Eukaryotes and prokaryotes use N-terminal destabilizing residues as a signal to target proteins for degradation by the N end rule pathway. In eukaryotes an E3 ligase, N-recognin, recognizes N-end rule substrates and mediates their ubiquitination and degradation by the proteasome. In Escherichia coli, N-end rule substrates are degraded by the AAA + chaperone ClpA in complex with the ClpP peptidase (ClpAP). Little is known of the molecular mechanism by which N-end rule substrates are initially selected for proteolysis. Here we report that the ClpAP-specific adaptor, ClpS, is essential for degradation of N-end rule substrates by ClpAP in bacteria. ClpS binds directly to N-terminal destabilizing residues through its substrate-binding site distal to the ClpS-ClpA interface, and targets these substrates to ClpAP for degradation. Degradation by the N-end rule pathway is more complex than anticipated and several other features are involved, including a net positive charge near the N terminus and an unstructured region between the N-terminal signal and the folded protein substrate. Through interaction with this signal, ClpS converts the ClpAP machine into a protease with exquisitely defined specificity, ideally suited to regulatory proteolysis. PMID- 16467844 TI - Gene expression during acute and prolonged hypoxia is regulated by distinct mechanisms of translational control. AB - Hypoxia has recently been shown to activate the endoplasmic reticulum kinase PERK, leading to phosphorylation of eIF2alpha and inhibition of mRNA translation initiation. Using a quantitative assay, we show that this inhibition exhibits a biphasic response mediated through two distinct pathways. The first occurs rapidly, reaching a maximum at 1-2 h and is due to phosphorylation of eIF2alpha. Continued hypoxic exposure activates a second, eIF2alpha-independent pathway that maintains repression of translation. This phase is characterized by disruption of eIF4F and sequestration of eIF4E by its inhibitor 4E-BP1 and transporter 4E-T. Quantitative RT-PCR analysis of polysomal RNA indicates that the translation efficiency of individual genes varies widely during hypoxia. Furthermore, the translation efficiency of individual genes is dynamic, changing dramatically during hypoxic exposure due to the initial phosphorylation and subsequent dephosphorylation of eIF2alpha. Together, our data indicate that acute and prolonged hypoxia regulates mRNA translation through distinct mechanisms, each with important contributions to hypoxic gene expression. PMID- 16467845 TI - Activated radixin is essential for GABAA receptor alpha5 subunit anchoring at the actin cytoskeleton. AB - Neurotransmitter receptor clustering is thought to represent a critical parameter for neuronal transmission. Little is known about the mechanisms that anchor and concentrate inhibitory neurotransmitter receptors in neurons. GABAA receptor (GABAAR) alpha5 subunits mainly locate at extrasynaptic sites and are thought to mediate tonic inhibition. Notably, similar as synaptic GABAARs, these receptor subtypes also appear in cluster formations at neuronal surface membranes and are of particular interest in cognitive processing. GABAAR alpha5 mutation or depletion facilitates trace fear conditioning or improves spatial learning in mice, respectively. Here, we identified the actin-binding protein radixin, a member of the ERM family, as the first directly interacting molecule that anchors GABAARs at cytoskeletal elements. Intramolecular activation of radixin is a functional prerequisite for GABAAR alpha5 subunit binding and both depletion of radixin expression as well as replacement of the radixin F-actin binding motif interferes with GABAAR alpha5 cluster formation. Our data suggest radixin to represent a critical factor in receptor localization and/or downstream signaling. PMID- 16467846 TI - Reduced meiotic crossovers and delayed prophase I progression in AtMLH3-deficient Arabidopsis. AB - Characterization of AtMLH3, the Arabidopsis homologue of the prokaryotic MutL mismatch repair gene, reveals that it is expressed in reproductive tissue where it is required for normal levels of meiotic crossovers (COs). Immunocytological studies in an Atmlh3 mutant indicate that chromosome pairing and synapsis proceed with normal distribution of the early recombination pathway proteins. Localization of the MutS homologue AtMSH4 occurs, suggesting that double Holliday junctions (dHjs) are formed, but the MutL homologue AtMLH1, which forms a heterocomplex with AtMLH3, fails to localize normally. Loss of AtMLH3 results in an approximately 60% reduction in COs and is accompanied by a substantial delay of approximately 25 h in prophase I progression. Analysis of the chiasma distribution in Atmlh3 suggests that dHj resolution can occur, but in contrast to wild type where most or all dHjs are directed to form COs the outcome is biased in favour of a non-CO outcome by a ratio of around 2 to 1. The data are compatible with a model whereby the MutL complex imposes a dHj conformation that ensures CO formation. PMID- 16467847 TI - Endocytic pathways regulate Toll-like receptor 4 signaling and link innate and adaptive immunity. AB - Immune responses are initiated when molecules of microbial origin are sensed by the Toll-like receptors (TLRs). We now report the identification of essential molecular components for the trafficking of the lipopolysaccharide (LPS) receptor complex. LPS was endocytosed by a receptor-mediated mechanism dependent on dynamin and clathrin and colocalized with TLR4 on early/sorting endosomes. TLR4 was ubiquitinated and associated with the ubiquitin-binding endosomal sorting protein hepatocyte growth factor-regulated tyrosine kinase substrate, Hrs. Inhibition of endocytosis and endosomal sorting increased LPS signaling. Finally, the LPS receptor complex was sorted to late endosomes/lysosomes for degradation and loading of associated antigens onto HLA class II molecules for presentation to CD4+ T cells. Our results show that endosomal trafficking of the LPS receptor complex is essential for signal termination and LPS-associated antigen presentation, thus controlling both innate and adaptive immunity through TLR4. PMID- 16467848 TI - Neuronal survival depends on EGFR signaling in cortical but not midbrain astrocytes. AB - Mice lacking epidermal growth factor receptor (EGFR) develop a neurodegeneration of unknown etiology affecting exclusively the frontal cortex and olfactory bulbs. Here, we show that EGFR signaling controls cortical degeneration by regulating cortical astrocyte apoptosis. Whereas EGFR(-/-) midbrain astrocytes are unaffected, mutant cortical astrocytes display increased apoptosis mediated by an Akt-caspase-dependent mechanism and are unable to support neuronal survival. The expression of many neurotrophic factors is unaltered in EGFR(-/-) cortical astrocytes suggesting that neuronal loss occurs as a consequence of increased astrocyte apoptosis rather than impaired secretion of trophic factors. Neuron specific expression of activated Ras can compensate for the deficiency of EGFR(-/ ) cortical astrocytes and prevent neuronal death. These results identify two functionally distinct astrocyte populations, which differentially depend on EGFR signaling for their survival and also for their ability to support neuronal survival. These spatial differences in astrocyte composition provide a mechanism for the region-specific neurodegeneration in EGFR(-/-) mice. PMID- 16467849 TI - Molecular chaperones and the assembly of the prion Sup35p, an in vitro study. AB - The protein Sup35 from Saccharomyces cerevisiae possesses prion properties. In vivo, a high molecular weight form of Sup35p is associated to the [PSI+] factor. The continued propagation of [PSI+] is highly dependent on the expression levels of molecular chaperones from the Hsp100, 70 and 40 families; however, so far, their role in this process is unclear. We have developed a reproducible in vitro system to study the effects of molecular chaperones on the assembly of full length Sup35p. We show that Hsp104p greatly stimulates the assembly of Sup35p into fibrils, whereas Ydj1p has inhibitory effect. Hsp82p, Ssa1p and Sis1p, individually, do not affect assembly. In contrast, Ssa1p together with either of its Hsp40 cochaperones blocks Sup35p polymerization. Furthermore, Ssa1p and Ydj1p or Sis1p can counteract the stimulatory activity of Hsp104p, by forming complexes with Sup35p oligomers, in an ATP-dependent manner. Our observations reveal the functional differences between Hsp104p and the Hsp70-40 systems in the assembly of Sup35p into fibrils and bring new insight into the mechanism by which molecular chaperones influence the propagation of [PSI+]. PMID- 16467850 TI - Vacuolar sequential exocytosis of large dense-core vesicles in adrenal medulla. AB - Individual exocytic events in intact adrenal medulla were visualized by two photon extracellular polar-tracer imaging. Exocytosis of chromaffin vesicles often occurred in a sequential manner, involving first vesicles located at the cell periphery and then those present deeper within the cytoplasm. Sequential exocytosis occurred preferentially at regions of the plasma membrane facing the intercellular space. The compound vesicles swelled to more than five times their original volume and formed vacuolar exocytic lumens as a result of expansion of intravesicular gels and their confinement within the lumen by the fusion pore and the narrow intercellular space. Such luminal swelling greatly promoted sequential exocytosis. The SNARE protein SNAP25 rapidly migrated from the plasma membrane to the membrane of fused vesicles. These data indicate that vesicles present deeper within the cytoplasm can be fusion ready like those at the cell periphery, and that swelling of exocytic lumens promotes assembly of the fusion machinery. We suggest the existence of two molecular configurations for fusion-ready states in Ca2+ -dependent exocytosis. PMID- 16467851 TI - Recruitment and activation of PLCgamma1 in T cells: a new insight into old domains. AB - Engagement of the T-cell antigen receptor leads to recruitment of phospholipase Cgamma1 (PLCgamma1) to the LAT-nucleated signaling complex and to PLCgamma1 activation in a tyrosine phosphorylation-dependent manner. The mechanism of PLCgamma1 recruitment and the role of PLCgamma1 Src homology (SH) domains in this process remain incompletely understood. Using a combination of biochemical methods and real-time fluorescent imaging, we show here that the N-terminal SH2 domain of PLCgamma1 is necessary but not sufficient for its recruitment. Either the SH3 or C-terminal SH2 domain of PLCgamma1, with the participation of Vav1, c Cbl and Slp76, are required to stabilize PLCgamma1 recruitment. All three PLCgamma1 SH domains are required for phosphorylation of PLCgamma1 Y783, which is critical for enzyme activation. These novel findings entailed revision of the currently accepted model of PLCgamma1 recruitment and activation in T lymphocytes. PMID- 16467852 TI - A hyper-dynamic equilibrium between promoter-bound and nucleoplasmic dimers controls NF-kappaB-dependent gene activity. AB - Because of its very high affinity for DNA, NF-kappaB is believed to make long lasting contacts with cognate sites and to be essential for the nucleation of very stable enhanceosomes. However, the kinetic properties of NF-kappaB interaction with cognate sites in vivo are unknown. Here, we show that in living cells NF-kappaB is immobilized onto high-affinity binding sites only transiently, and that complete NF-kappaB turnover on active chromatin occurs in less than 30 s. Therefore, promoter-bound NF-kappaB is in dynamic equilibrium with nucleoplasmic dimers; promoter occupancy and transcriptional activity oscillate synchronously with nucleoplasmic NF-kappaB and independently of promoter occupancy by other sequence-specific transcription factors. These data indicate that changes in the nuclear concentration of NF-kappaB directly impact on promoter function and that promoters sample nucleoplasmic levels of NF-kappaB over a timescale of seconds, thus rapidly re-tuning their activity. We propose a revision of the enhanceosome concept in this dynamic framework. PMID- 16467853 TI - Subtelomeric factors antagonize telomere anchoring and Tel1-independent telomere length regulation. AB - Yeast telomeres are anchored at the nuclear envelope (NE) through redundant pathways that require the telomere-binding factors yKu and Sir4. Significant variation is observed in the efficiency with which different telomeres are anchored, however, suggesting that other forces influence this interaction. Here, we show that subtelomeric elements and the insulator factors that bind them antagonize the association of telomeres with the NE. This is detectable when the redundancy in anchoring pathways is compromised. Remarkably, these same conditions lead to a reduction in steady-state telomere length in the absence of the ATM-kinase homologue Tel1. Both the delocalization of telomeres and reduction in telomere length can be induced by targeting of Tbf1 or Reb1, or the viral transactivator VP16, to a site 23 kb away from the TG repeat. This correlation suggests that telomere anchoring and a Tel1-independent pathway of telomere length regulation are linked, lending a functional significance to the association of yeast telomeres with the NE. PMID- 16467854 TI - Subtelomeric proteins negatively regulate telomere elongation in budding yeast. AB - The Tbf1 and Reb1 proteins are present in yeast subtelomeric regions. We establish in this work that they inhibit telomerase-dependent lengthening of telomere. For example, tethering the N-terminal domain of Tbf1 and Reb1 in a subtelomeric region shortens that telomere proportionally to the number of domains bound. We further identified a 90 amino-acid long sequence within the N terminal domain of Tbf1 that is necessary but not sufficient for its length regulation properties. The role of the subtelomeric factors in telomere length regulation is antagonized by TEL1 and does not correlate with a global telomere derepression. We show that the absence of TEL1 induces an alteration in the structure of telomeric chromatin, as defined biochemically by an increased susceptibility to nucleases and a greater heterogeneity of products. We propose that the absence of TEL1 modifies the organization of the telomeres, which allows Tbf1 and Reb1 to cis-inhibit telomerase. The involvement of subtelomeric factors in telomere length regulation provides a possible mechanism for the chromosome specific length setting observed at yeast and human telomeres. PMID- 16467856 TI - Monochorionic monoamniotic twins: neonatal outcome. AB - BACKGROUND: Monochorionic monoamniotic twins (MoMo) occur in one of 10,000 pregnancies. Cord entanglement, malformations, twin-to-twin transfusion syndrome (TTS) and prematurity are responsible for their high perinatal morbidity and mortality. OBJECTIVE: To report our experience with 36 sets of MoMo twins (1990 to 2005) and to provide updated information for counseling. METHODS: Chorionicity was determined by placental examination, gestational age and TTS clinically and by sonography. Intrauterine growth restriction (IUGR) was diagnosed with a twin specific nomogram. RESULTS: Cord entanglement was observed in 15 pregnancies, but only one twin with entanglement and a true knot, experienced related morbidity. Four of 71 live births were IUGR. Malformations were diagnosed prenatally (one hypoplastic left heart and one body stalk) and postnatally (one vertebral anomalies-anal atresia-tracheoesophageal fistula-renal defect (VATER) and two lung hypoplasias). Twin-to-twin transfusion syndrome affected three sets of twins. Five twin sets delivered before 31, 19 sets at 31 to 32 and 12 sets at 33 to 34 weeks. Six of 71 (8%) twins died (four malformations, one TTS and one 26 weeks premature). Head ultrasounds in 59 of 65 survivors showed two (3%) periventricular leukomalacia, five (9%) Grade I-II intraventricular hemorrhage and 52 (88%) normal. CONCLUSIONS: Monochorionic monoamniotic twins remain a group at risk for cord entanglement, congenital malformations, TTS and prematurity. Although their neonatal mortality and morbidity is high, outcomes for survival are better than anticipated. PMID- 16467855 TI - Structure of glutamate carboxypeptidase II, a drug target in neuronal damage and prostate cancer. AB - Membrane-bound glutamate carboxypeptidase II (GCPII) is a zinc metalloenzyme that catalyzes the hydrolysis of the neurotransmitter N-acetyl-L-aspartyl-L-glutamate (NAAG) to N-acetyl-L-aspartate and L-glutamate (which is itself a neurotransmitter). Potent and selective GCPII inhibitors have been shown to decrease brain glutamate and provide neuroprotection in preclinical models of stroke, amyotrophic lateral sclerosis, and neuropathic pain. Here, we report crystal structures of the extracellular part of GCPII in complex with both potent and weak inhibitors and with glutamate, the product of the enzyme's hydrolysis reaction, at 2.0, 2.4, and 2.2 A resolution, respectively. GCPII folds into three domains: protease-like, apical, and C-terminal. All three participate in substrate binding, with two of them directly involved in C-terminal glutamate recognition. One of the carbohydrate moieties of the enzyme is essential for homodimer formation of GCPII. The three-dimensional structures presented here reveal an induced-fit substrate-binding mode of this key enzyme and provide essential information for the design of GCPII inhibitors useful in the treatment of neuronal diseases and prostate cancer. PMID- 16467857 TI - Antenatal betamethasone treatment has a persisting influence on infant HPA axis regulation. AB - OBJECTIVE: To examine the consequences of antenatal betamethasone (AB) exposure on postnatal stress regulation. STUDY DESIGN: Fourteen AB exposed infants born at 28-30 weeks' gestation were assessed in the NICU during postnatal week 1 and at 34 weeks postconception. Nine infants born at 34 weeks gestation without AB treatment were evaluated as a postconceptional age comparison group. Salivary cortisol, heart rate, and behavior were measured at baseline and in response to a heelstick blood draw. RESULTS: Repeated measures ANOVAs revealed that both groups displayed an increase in heart rate and behavioral distress in response to the stressor. The cortisol response, however, was blunted in AB-treated infants at both assessments. CONCLUSION: AB treatment has consequences for hypothalamic pituitary-adrenal (HPA) axis regulation that persist for at least four to six weeks after birth, indicating that studies of long-term effects are warranted. PMID- 16467858 TI - Brain activation by visual erotic stimuli in healthy middle aged males. AB - The objective of the present study was to identify brain centers, whose activity changes are related to erotic visual stimuli in healthy, heterosexual, middle aged males. Ten heterosexual, right-handed males with normal sexual function were entered into the present study (mean age 52 years, range 46-55). All potential subjects were screened over 1 h interview, and were encouraged to fill out questionnaires including the Brief Male Sexual Function Inventory. All subjects with a history of sexual arousal disorder or erectile dysfunction were excluded. We performed functional brain magnetic resonance imaging (fMRI) in male volunteers when an alternatively combined erotic and nonerotic film was played for 14 min and 9 s. The major areas of activation associated with sexual arousal to visual stimuli were occipitotemporal area, anterior cingulate gyrus, insula, orbitofrontal cortex, caudate nucleus. However, hypothalamus and thalamus were not activated. We suggest that the nonactivation of hypothalamus and thalamus in middle aged males may be responsible for the lesser physiological arousal in response to the erotic visual stimuli. PMID- 16467859 TI - A novel chimeric promoter that is highly responsive to hypoxia and metals. AB - To develop a potent hypoxia-inducible promoter, we evaluated the usefulness of chimeric combinations of the (Egr-1)-binding site (EBS) from the Egr-1 gene, the metal-response element (MRE) from the metallothionein gene, and the hypoxia response element (HRE) from the phosphoglycerate kinase 1 gene. In transient transfection assays, combining three copies of HRE (3 x HRE) with either EBS or MRE significantly increased hypoxia responsiveness. When a three-enhancer combination was tested, the EBS-MRE-3 x HRE (E-M-H) gave a hypoxia induction ratio of 69. The expression induced from E-M-H-pGL3 was 2.4-fold higher than that induced from H-pGL3 and even surpassed the expression from a human cytomegalovirus promoter-driven vector. The high inducibility of E-M-H was confirmed by validation studies in different cells and by expressing other cDNAs. Gel shift assays together with functional overexpression studies suggested that increased levels of hypoxia-inducible factor 1alpha, metal transcription factor-1 and Egr-1 may be associated with the high inducibility of the E-M-H chimeric promoter. E-M-H was also induced by hypoxia mimetics such as Co2+ and deferoxamine (DFX) and by hydrogen peroxide. Gene expression from the E-M-H was reversible as shown by the reduced expression of the transgene upon removal of inducers such as hypoxia and DFX. In vivo evaluation of the E-M-H in ischemic muscle revealed that erythropoietin secretion and luciferase and LacZ expression were significantly higher in the E-M-H group than in a control or H group. With its high induction capacity and versatile means of modulation, this novel chimeric promoter should find wide application in the treatment of ischemic diseases and cancer. PMID- 16467860 TI - Sustained expression after nonviral ocular gene transfer using mammalian promoters. AB - The CMV promoter drives high transgene expression and is one of the most commonly used promoters for gene transfer. Tissue-specific mammalian promoters provide an alternative, and it would be useful to have a system to directly compare them to viral promoters free from potential confounding vector-related effects. In this study, we describe how electroporation after subretinal injection of plasmid DNA can be used to perform comparative quantitative analysis of promoter activities. Luciferase assay of eyecup homogenates was carried out after coinjection/electroporation of pGL2, a plasmid containing the promoter fragment of interest coupled to the firefly luciferase gene, and pRL-CMV, a plasmid containing the CMV promoter coupled to the Renilla luciferase gene for normalization. This technique was used to compare activity of different fragments of the 5'-upstream region of the vitelliform macular dystrophy 2 (VMD2) gene, which is selectively expressed in the retinal pigmented epithelial (RPE) cells, and results indicated positive regulatory elements between -104 and -154 bp and between -424 and -585 bp. Addition of a fragment from intron 1 reduced the activity of the -585/+38 bp fragment by 75%. Deletion analysis implicated a 342 bp region near the 5'-end of intron 1 in the repression. Results of transient transfections in two cell lines that constitutively express VMD2 were similar, and results in transgenic mice were consistent, providing validation for promoter analysis by in vivo electroporation. We then explored the time course of expression of the -585/+38 VMD2 promoter fragment and found that compared to cassettes driven by CMV or SV40 promoters, which showed peak luciferase activity on day 2 followed by a rapid decrease in activity, the VMD2 promoter fragment showed lower activity initially, but the activity was sustained for up to 56 days (longest time point measured). A promoter fragment from another RPE-specific gene, Rpe65, showed a similar pattern of sustained expression for at least 112 days. These data indicate that nonviral gene transfer can be used to quantitatively evaluate the activity of promoter fragments independent of influence from viral vectors. A potentially important finding using this new technique is the demonstration that relatively sustained passenger gene expression can be achieved with nonviral gene transfer using mammalian rather than viral promoters. PMID- 16467861 TI - Reactive oxygen species production by circulating monocytes: insights from pathophysiology to clinical hypertension. PMID- 16467862 TI - Carotid artery intima-media thickness and reactive oxygen species formation by monocytes in hypertensive patients. AB - Carotid artery intima-media thickness (IMT) is a widely accepted index for assessing atherosclerosis, and is known to be a risk indicator for cardiovascular and cerebrovascular events. Oxidative stress and inflammation are also known to play critical roles in the pathogenesis of vascular events. We studied the association between IMT and inflammatory markers, such as oxidative stress in polymorphonuclear leukocytes (PMNs) and mononuclear cells (MNCs) in 156 patients with essential hypertension. Reactive oxygen species (ROS) formation by PMNs and MNCs was measured by gated flow cytometry. CRP and traditional risk factors, such as age, gender, body mass index, hemoglobin A(1c), and total cholesterol were also measured. The subjects were divided into a plaque group (max-IMT>or=1.1 mm, n=40), and a nonplaque group (max-IMT<1.1 mm, n=116). ROS formation by MNCs was significantly increased in the plaque group when compared with the nonplaque group (P<0.0001). Multiple regression analysis revealed a significant correlation between IMT and ROS formation by MNCs (r=0.407, P<0.0001), or CRP (r=0.216, P=0.0029) or hemoglobinA1c (r=0.158, P=0.0270) or age (r=0.157, P=0.0447). No significant correlation was observed between IMT and ROS formation by PMNs. These results suggest that carotid artery IMT may be affected by increased ROS formation by MNCs, and that increased ROS formation by MNCs may be related to the development of atherosclerosis. We propose that ROS formation by MNCs is a marker for prediction of carotid atherosclerosis. PMID- 16467863 TI - The presence of a BCR-ABL mutant allele in CML does not always explain clinical resistance to imatinib. AB - The expansion of a leukemia clone bearing a Bcr-Abl kinase domain mutation is associated with acquired resistance to imatinib and may also predict disease progression in patients with Philadelphia-positive chronic myeloid leukemia (CML). Here we report results of pyrosequencing to quantitate the non-mutated and mutant alleles in 12 CML patients monitored over periods ranging from 11 to 58 months, and describe three contrasting kinetic patterns: Group 1 - in four patients total BCR-ABL transcript numbers remained high with the mutant allele predominating; Group 2 - in four patients the total number of BCR-ABL transcripts fell to low levels but the mutant allele predominated; and Group 3 - in four other patients the total level of transcripts remained high (n = 2) or fell (n = 2) but the mutant clone persisted at relatively low level. In Group 2 the mutant leukemia clone was presumably still relatively sensitive to imatinib but in Group 1 the leukemia could be classified as resistant. In Group 3 patients the imatinib sensitivity of the leukemia was variable. We conclude that a mutant clone does not necessarily have a proliferative advantage and its presence does not always account for resistance to imatinib. Other mechanisms underlie resistance in at least some patients. PMID- 16467864 TI - Hyperactivation of the RAS signaling pathway in myelodysplastic syndrome with AML1/RUNX1 point mutations. AB - AML1/RUNX1 mutations have been reported frequently in myelodysplastic syndrome (MDS) patients, especially those diagnosed with refractory anemia with excess blast (RAEB), RAEB in transformation (RAEBt), or AML following MDS (these categories are defined as MDS/AML). Although AML1 mutations are suspected to play a pivotal role in the development of MDS/AML, acquisition of additional genetic alterations is also necessary. We analyzed gene alterations in MDS/AML patients with AML1 mutations, comparing them to alterations in those without an AML1 mutation. AML1 mutations were significantly associated with -7/7q-, whereas MDS/AML patients without AML1 mutations showed a high frequency of -5/5q- and a complex karyotype. Patients with AML1 mutations showed more mutations of their FLT3, N-RAS, PTPN11, and NF1 genes, resulting in a significantly higher mutation frequency for receptor tyrosine kinase (RTK)-RAS signaling pathways in AML1 mutated MDS/AML patients compared to AML1-wild-type MDS/AML patients (38% versus 6.3%, P < 0.0001). Conversely, p53 mutations were detected only in patients without AML1 mutations. Furthermore, blast cells of the AML1-mutated patients expressing surface c-KIT, and SHP-2 mutants contributed to prolonged and enhanced extracellular signal-regulated kinase activation following stem cell factor stimulation. Our results suggest that MDS/AML arising from AML1/RUNX1 mutations has a significant association with -7/7q- alteration, and frequently involves RTK RAS signaling pathway activation. PMID- 16467865 TI - High frequency of several PIG-A mutations in patients with aplastic anemia and myelodysplastic syndrome. AB - To clarify some characteristics of phosphatidylinositol glycan-class A gene (PIG A) mutations in aplastic anemia (AA) and myelodysplastic syndrome (MDS) patients compared with those in paroxysmal nocturnal hemoglobinuria (PNH) patients, we investigated PIG-A mutations in CD59- granulocytes and CD48- monocytes from seven AA, eight MDS, and 11 PNH Japanese patients. The most frequent base or type abnormalities of the PIG-A gene in AA and MDS patients were base substitutions or missense mutations, respectively, and deletions or frameshift mutations, respectively, in PNH patients. Several PIG-A mutations, most of which were statistically minor, were found in glycosylphosphatidylinositol-negative cells from all AA and MDS patients but not from all PNH patients. However, the common PIG-A mutations during the clinical course between CD59- granulocytes and/or CD48 monocytes from each AA or MDS patient, except for Case 5, were not found. PIG-A mutations were different between the granulocytes and monocytes from five AA and five MDS patients. Our results indicate that there were some characteristics of PIG-A mutations in AA and MDS patients compared with PNH patients and that several minor PNH clones in these patients occurred at random during the clinical course. This partly explains the transformation of AA or MDS to PNH at intervals. PMID- 16467866 TI - Hyperforin inhibits MMP-9 secretion by B-CLL cells and microtubule formation by endothelial cells. AB - We previously reported that hyperforin (HF), a natural phloroglucinol purified from Saint John's wort, can induce the apoptosis of leukemic cells from patients with B-cell lymphocytic leukemia (B-CLL) ex vivo. We show here that treatment of cultured B-CLL patients' cells with HF results in a marked inhibition of their capacity to secrete matrix metalloproteinase-9, an essential component in neo angiogenesis through degradation of the extracellular matrix process. The phloroglucinol acts by decreasing the production of the latent 92 kDa pro-enzyme. The inhibitory effect of HF is associated with a decrease in VEGF release by the leukemic cells. Moreover, HF is found to prevent the formation of microtubules by human bone marrow endothelial cells cultured on Matrigel, evidencing its capacity to inhibit vessel formation. Our results show the antiangiogenesis activity of HF and strengthen its potential interest in the therapy of B-CLL. PMID- 16467867 TI - Preferential expression of a high number of ATP binding cassette transporters in both normal and leukemic CD34+CD38- cells. PMID- 16467868 TI - NUP98 rearrangements in hematopoietic malignancies: a study of the Groupe Francophone de Cytogenetique Hematologique. AB - The NUP98 gene is fused with 19 different partner genes in various human hematopoietic malignancies. In order to gain additional clinico-hematological data and to identify new partners of NUP98, the Groupe Francophone de Cytogenetique Hematologique (GFCH) collected cases of hematological malignancies where a 11p15 rearrangement was detected. Fluorescence in situ hybridization (FISH) analysis showed that 35% of these patients (23/66) carried a rearrangement of the NUP98 locus. Genes of the HOXA cluster and the nuclear-receptor set domain (NSD) genes were frequently fused to NUP98, mainly in de novo myeloid malignancies whereas the DDX10 and TOP1 genes were equally rearranged in de novo and in therapy-related myeloid proliferations. Involvement of ADD3 and C6ORF80 genes were detected, respectively, in myeloid disorders and in T-cell acute lymphoblastic leukemia (T-ALL), whereas the RAP1GDS1 gene was fused to NUP98 in T ALL. Three new chromosomal breakpoints: 3q22.1, 7p15 (in a localization distinct from the HOXA locus) and Xq28 were detected in rearrangements with the NUP98 gene locus. The present study as well as a review of the 73 cases previously reported in the literature allowed us to delineate some chromosomal, clinical and molecular features of patients carrying a NUP98 gene rearrangements. PMID- 16467870 TI - GATA-3 promotes Th2 responses through three different mechanisms: induction of Th2 cytokine production, selective growth of Th2 cells and inhibition of Th1 cell specific factors. AB - Naive CD4 T cells can differentiate into at least two different types of T helpers, Th1 and Th2 cells. Th2 cells, capable of producing IL-4, IL-5 and IL-13, are involved in humoral immunity against extracellular pathogens and in the induction of asthma and other allergic diseases. In this review, we summarize recent reports regarding the transcription factors involved in Th2 differentiation and cell expansion, including Stat5, Gfi-1 and GATA-3. Stat5 activation is necessary and sufficient for IL-2-mediated function in Th2 differentiation. Enhanced Stat5 signaling induces Th2 differentiation independent of IL-4 signaling; although it does not up-regulate GATA-3 expression, it does require the presence of GATA-3 for its action. Gfi-1, induced by IL-4, promotes the expansion of GATA-3-expressing cells. Analysis of conditional Gata3 knockout mice confirmed the critical role of GATA-3 in Th2 cell differentiation (both IL-4 dependent and IL-4 independent) and in Th2 cell proliferation and also showed the importance of basal GATA-3 expression in inhibiting Th1 differentiation. PMID- 16467871 TI - The quantal theory of immunity. AB - Exactly how the immune system discriminates between all environmental antigens to which it reacts vs. all self-antigens to which it does not, is a principal unanswered question in immunology. As set forth in this review, because of the advances in our understanding of the immune system that have occurred in the last 50 years, for the first time it is possible to formulate a new theory, termed the "Quantal Theory of Immunity", which reduces the problem from the immune system as a whole, to the individual cells comprising the system, and finally to a molecular explanation as to how the system behaves as it does. PMID- 16467872 TI - Cytokines as critical co-stimulatory molecules in modulating the immune response of natural killer cells. AB - Cytokines are involved in directing the activation of natural killer (NK) cells. NK cells are involved in the recognition of cells that have been altered; thus they do not recognize specific insults to the host, but when activated, are capable of destroying infected cells directly, as well as promoting the recruitment and response of the other components of the immune system by the release of cytokines and chemokines. It is these properties that have made NK cells a critical part of innate immunity and adaptive immunity, and they play a principal role linking innate and adaptive immunity by the recruitment of an adaptive immune response to an innate immune reaction. PMID- 16467873 TI - Targeting gene-virotherapy of cancer. AB - Our purpose is to completely elimination of xenograft tumor in animal tumor model in order to work out a protocal for the cure of patient. Gene therapy and viral therapy for cancer have got some therapeutic effects, but both have no great breakthrough. Therefore, we worked out a new strategy called Targeting Gene Virotherapy of Cancer which is a combination of the advantage of gene therapy and virotherapy. This new strategy has stronger antitumor effect than either of them alone. A tumor specific replicative adenovirus vector ZD55 (E1B 55KD deleted Adv.) which is similar to ONYX-015 in targeting fuction but significant different in construction was produced and various single therapeutic gene was inserted into ZD55. Now such a conception as Targeting Gene-Virotherapy of Cancer was raised and systemically studied before, although there are some works on ONYX-015 tk, -cd or cd/-tk etc. separately. The antitumor effect of ZD55-Gene (for example IL-24 gene) is much better than ZD55 (virotherapy) alone and hundred fold high than that of Ad-IL-24 (gene therapy) alone. ZD55-IL-24 was in preclinal studying in the ZD55-IL-24 therapy, completely elimination of tumor mass was occurred in some mice but not in all mice, that means one gene was not effictive enough to eliminate all the tumor mass in all mice. Therefore two genes with compensative or synergetic effect were inserted into ZD55 separately and used in combination. This strategy was called Targeting Dual Gene-Virotherapy of Cancer (with PCT patent). Then much better results were obtained and all the xenograft tumor masses were completely eliminated in all mice, if two suitable genes were chosen. On the basis of the initiation of two gene results, it was thought about that using two tumors promoter to control the virus vector will be better for the targeting effect and the safty of the drugs. Then double tumor controlled virus vector harboring two genes for cancer therapy was worked out. Better results have been obtained and another patent has been applied. This antitumor strategy could be used to kill all the tumor cells completely in all mice with minimum damage to normal cells. PMID- 16467874 TI - Approaches to functional genomics in filamentous fungi. AB - The study of gene function in filamentous fungi is a field of research that has made great advances in very recent years. A number of transformation and gene manipulation strategies have been developed and applied to a diverse and rapidly expanding list of economically important filamentous fungi and oomycetes. With the significant number of fungal genomes now sequenced or being sequenced, functional genomics promises to uncover a great deal of new information in coming years. This review discusses recent advances that have been made in examining gene function in filamentous fungi and describes the advantages and limitations of the different approaches. PMID- 16467875 TI - The role of NBS1 in DNA double strand break repair, telomere stability, and cell cycle checkpoint control. AB - The genomes of eukaryotic cells are under continuous assault by environmental agents and endogenous metabolic byproducts. Damage induced in DNA usually leads to a cascade of cellular events, the DNA damage response. Failure of the DNA damage response can lead to development of malignancy by reducing the efficiency and fidelity of DNA repair. The NBS1 protein is a component of the MRE11/RAD50/NBS1 complex (MRN) that plays a critical role in the cellular response to DNA damage and the maintenance of chromosomal integrity. Mutations in the NBS1 gene are responsible for Nijmegen breakage syndrome (NBS), a hereditary disorder that imparts an increased predisposition to development of malignancy. The phenotypic characteristics of cells isolated from NBS patients point to a deficiency in the repair of DNA double strand breaks. Here, we review the current knowledge of the role of NBS1 in the DNA damage response. Emphasis is placed on the role of NBS1 in the DNA double strand repair, modulation of the DNA damage sensing and signaling, cell cycle checkpoint control and maintenance of telomere stability. PMID- 16467876 TI - Preassembly and ligand-induced restructuring of the chains of the IFN-gamma receptor complex: the roles of Jak kinases, Stat1 and the receptor chains. AB - We previously demonstrated using noninvasive technologies that the interferon gamma (IFN-gamma) receptor complex is preassembled (1). In this report we determined how the receptor complex is preassembled and how the ligand-mediated conformational changes occur. The interaction of Stat1 with IFN-gammaR1 results in a conformational change localized to IFN-gammaR1. Jak1 but not Jak2 is required for the two chains of the IFN-gamma receptor complex (IFN-gammaR1 and IFN-gammaR2) to interact; however, the presence of both Jak1 and Jak2 is required to see any ligand-dependant conformational change. Two IFN-gammaR2 chains interact through species-specific determinants in their extracellular domains. Finally, these determinants also participate in the interaction of IFN-gammaR2 with IFN-gammaR1. These results agree with a detailed model of the IFN-gamma receptor that requires the receptor chains to be pre-associated constitutively for the receptor to be active. PMID- 16467877 TI - Inflammatory bowel disease requires the interplay between innate and adaptive immune signals. AB - Inflammatory bowl disease (IBD) is a type 1 T helper cell (Th1)-mediated autoimmune disease. Various studies have revealed that environmental pathogens also play a significant role in the initiation and progression of this disease. Interestingly, the pathogenesis of IBD has been shown to be related to nitric oxide (NO) released from innate immune cells. Although NO is known to be highly toxic to the gut epithelia, there is very little information about the regulation of NO production, One major question in the etiology of IBD is how Th1 cells and pathogens interact in the induction of IBD. In present study, we focused on the regulation of NO. We show that macrophages require both interferon-gamma (IFN gamma)-mediated and TLR4-mediated signals for the production of NO, which causes inflammation in the intestine and subsequently IBD. Thus, IBD is the result of concerted actions of innate immune signals, such as the binding of LPS to TLR-4, and adaptive immune signals, such as IFN-gamma produced by Th1 cells. PMID- 16467878 TI - uPAR expression under hypoxic conditions depends on iNOS modulated ERK phosphorylation in the MDA-MB-231 breast carcinoma cell line. AB - Urokinase plasminogen activator receptor (uPAR) plays a major role in cancer invasion and metastasis and uPAR expression is correlated with a poor prognosis in various cancer types. Moreover, the expression of uPAR is increased under hypoxic conditions. Nitric oxide (NO) and its metabolites produced by inducible nitric oxide synthase (iNOS) are important products of hypoxic stress, and NO may activate or modulate extracellular signal regulated kinase (ERK). Here, we evaluated uPA, uPAR, and activated ERK levels under hypoxic conditions, and the modulatory effects of iNOS and NO in the MDA-MB-231 human breast cancer cell line. Cells were incubated in a hypoxic or normoxic incubator and treated with PD98059 (a MEK 1/2 inhibitor, which abrogates ERK phosphorylation) and aminoguanidine (a selective iNOS inhibitor). uPAR expression, ERK phosphorylation, and uPA activity were found to be increased under hypoxic conditions. Moreover, when cells were treated with PD98059 under hypoxic conditions, uPAR was downregulated, whereas aminoguanidine markedly increased ERK phosphorylation in a dose dependent manner. Furthermore, aminoguanidine increased uPAR expression and prevented the inhibition of uPAR expression by PD98059. These results demonstrated that uPAR is induced by hypoxia and that increased uPAR expression is mediated by ERK phosphorylation, which in turn is modulated by iNOS/NO in MDA-MB-231 cells. We conclude that iNOS/NO downregulates the expression of uPAR under hypoxic conditions via ERK pathway modulation. PMID- 16467879 TI - Blocking of N-acetylglucosaminyltransferase V induces cellular endoplasmic reticulum stress in human hepatocarcinoma 7,721 cells. AB - N-acetylglucosaminyltransferase V (GnT-V) is an important tumorigenesis and metastasis-associated enzyme. To study its biofunction, the GnT-V stably suppressed cell line (GnT-V-AS/7,721) was constructed from 7,721 hepatocarcinoma cells in previous study. In this study, cDNA array gene expression profiles were compared between GnT-V-AS/7,721 and parental 7,721 cells. The data indicated that GnT-V-AS/7,721 showed a characteristic expression pattern consistent with the ER stress. The molecular mechanism of the ER stress was explored in GnT-V-AS/7,721 by the analysis on key molecules in both two unfolded protein response (UPR) pathways. For ATF6 and Ire1/XBP-1 pathway, it was evidenced by the up-regulation of BIP at mRNA and protein level, and the appearance of the spliced form of XBP 1. As for PERK/eIF2alpha pathway, the activation of ER eIF2alpha kinase PERK was observed. To confirm the results from GnT-V-AS/7,721 cells, the key molecules in the UPR were examined again in 7,721 cells interfered with the GnT-V by the specific RNAi treatment. The results were similar with those from GnT-V-AS/7721, indicating that blocking of GnT-V can specifically activate ER stress in 7,721 cells. Rate of (3)H-Man incorporation corrected with rate of (3)H-Leu incorporation in GnT-V-AS/7,721 was down-regulated greatly compared with the control, which demonstrated the deficient function of the enzyme synthesizing N glycans after GnT-V blocking. Moreover, the faster migrating form of chaperone GRP94 associated with the underglycosylation, and the extensively changed N glycans structures of intracellular glycoproteins were also detected in GnT-V AS/7,721. These results supported the mechanism that blocking of GnT-V expression impaired functions of chaperones and N-glycan-synthesizing enzymes, which caused UPR in vivo. PMID- 16467880 TI - Phosphorylated KDR can be located in the nucleus of neoplastic cells. AB - KDR (kinase insert domain receptor) phosphorylation induces several effects which lead eventually to cell proliferation and survival. The precise mechanisms by which KDR, once it is activated, communicates with the nucleus are starting to be understood but have not yet been completely unravelled. Two in vitro studies on animal cell lines reported in the literature have demonstrated that, following stimulation with VEGF, KDR is actually translocated within the nucleus. Our aim was to investigate whether this translocation occurs in human cells both in vitro and in vivo. Using laser scanning confocal microscopy, a variable nuclear localization of phosphorylated and total KDR in cell lines and tumour samples was found. In human neoplastic cell lines, hypoxic stimulation greatly increased the nuclear amount of total KDR but less so that of the phosphorylated form. Only after hypoxia and VEGF stimulation there was a comparably increased expression of phosphorylated and total KDR observed in the nuclei of these cells. We conclude that neoplastic cells show a variable expression of total and phosphorylated KDR in the nucleus. The precise functional meaning of nuclear location remains to be established. PMID- 16467881 TI - Gene silencing in Xenopus laevis by DNA vector-based RNA interference and transgenesis. AB - A vector-based RNAi expression system was developed using the Xenopus tropicalis U6 promoter, which transcribes small RNA genes by RNA polymerase III. The system was first validated in a Xenopus laevis cell line, designing a short hairpin DNA specific for the GFP gene. Co-transfection of the vector-based RNAi and the GFP gene into Xenopus XR1 cells significantly decreased the number of GFP-expressing cells and overall GFP fluorescence. Vector-based RNAi was subsequently validated in GFP transgenic Xenopus embryos. Sperm nuclei from GFP transgenic males and RNAi construct-incubated-sperm nuclei were used for fertilization, respectively. GFP mRNA and protein were reduced by approximately 60% by RNAi in these transgenic embryos compared with the control. This transgene-driven RNAi is specific and stable in inhibiting GFP expression in the Xenopus laevis transgenic line. Gene silencing by vector-based RNAi and Xenopus transgenesis may provide an alternative for 'repression of gene function' studies in vertebrate model systems. PMID- 16467882 TI - The identification of a new actin-binding region in p57. AB - The actin-binding protein p57 is a member of mammalian coronin-like proteins. The roles of this protein in phagocytic processes conceivably depend on its interactions with F-actin. Two regions, p57(1-34) and p57(111-204), were previously reported to be actin-binding sites. In this study, we found that the C terminal region of p57, p57(297-461), also possessed F-actin binding activity. Furthermore, the leucine zipper domain at the C-terminus of p57(297-461) was essential for this actin-binding activity. The F-actin cross-linking assay revealed that the region contained in p57(297-461) was sufficient to cross-link actin filaments. Our results strongly suggested that there was a new actin binding region at the C-terminus of p57. PMID- 16467883 TI - Modulation of the activation of Stat1 by the interferon-gamma receptor complex. AB - The activation of Stat1 by the interferon-gamma (IFN-gamma) receptor complex is responsible for the transcription of a significant portion of IFN-gamma induced genes. Many of these genes are responsible for the induction of an apoptotic state in response to IFN-gamma. In the absence of Stat1 activation, IFN-gamma instead induces a proliferative response. Modifying Stat1 activation by IFN-gamma may have pharmacological benefits. We report that the rate of activation of Stat1 can be altered in HeLa cells by overexpressing either the IFN-gammaR1 chain or the IFN-gammaR2 chain. These alterations occur in hematopoietic cell lines: Raji cells and monocytic cell lines, which have average and above-average IFN-gammaR2 surface expression, activate Stat1 similarly to HeLa cells and HeLa cells overexpressing IFNgammaR2, respectively. The rapid Stat1 activation seen in HeLa cells can be inhibited by overexpressing a chimeric IFN-gammaR2 chain that does not bind Jak2 or (when high concentrations of IFN-gamma are used) by overexpressing IFN-gammaR1. These data are consistent with a model in which the recruitment of additional Jak2 activity to a signaling complex accelerates the rate of Stat1 activation. We conclude that the rate of activation of Stat1 in cells by IFN-gamma can be modified by regulating either receptor chain and speculate that pharmacological agents which modify receptor chain expression may alter IFN-gamma receptor signal transduction. PMID- 16467886 TI - Unanswered questions in nephrology. PMID- 16467884 TI - Infection, immune responses and the aetiology of childhood leukaemia. AB - Childhood leukaemia is the principal subtype of paediatric cancer and, despite success in treatment, its causes remain enigmatic. A plethora of candidate environmental exposures have been proposed, but most lack a biological rationale or consistent epidemiological evidence. Although there might not be a single or exclusive cause, an abnormal immune response to common infection(s) has emerged as a plausible aetiological mechanism. PMID- 16467887 TI - Hemoperitoneum in a woman with acute paraplegia. PMID- 16467888 TI - Is homocysteine an iron-dependent cardiovascular risk factor? AB - Recent studies of endothelial function suggest that adverse vascular effects of homocysteine are iron-dependent. Iron sucrose worsens and iron chelation eliminates homocysteine-associated decreases in flow-mediated dialation. There may be no vasculopathic effect of homocysteine without available reactive iron. Iron-dependent amplification of the vascular effects of homocysteine may be one of several mechanisms by which stored iron increases cardiac risk. PMID- 16467889 TI - Fifty years of Balkan endemic nephropathy: daunting questions, elusive answers. AB - Balkan endemic nephropathy (BEN) has remained a geographically constant endemic for 50 years. Despite extensive research, its etiology remains unknown. In the current issue, in a study in one of the earliest sites where the endemic was first recognized, Dimitrov et al. confirm the persistance of the endemic into a new generation and also identify a maternal link in the pathogenesis of BEN. This intriguing finding needs to be confirmed in other endemic areas. PMID- 16467893 TI - Energetics from slow infrared multiphoton dissociation of biomolecules. AB - Photodissociation kinetics of the protonated pentapeptide leucine enkephalin measured using a cw CO(2) laser and a Fourier-transform mass spectrometer are reported. A short induction period, corresponding to the time required to raise the internal energy of the ion population to a (dissociating) steady state, is observed. After this induction period, the dissociation data are accurately fit by first-order kinetics. A plot of the log of the unimolecular dissociation rate constant, k(uni), as a function of the log of laser power is linear at low laser powers (<9 W, k(uni) <0.05 s(-1)), but tapers off at high laser power (9-33 W, k(uni) = 0.05-7 s(-1)). The entire measured dissociation curve can be accurately fit by an exponential function plus a constant. The experiment is simulated using a master equation formalism. In the model, the laser radiation is described as an energetically flat-topped distribution which is spatially uniform. This description is consistent with experimental results which indicate that ion motion within the cell averages out spatial inhomogeneities in the laser light. The model has several adjustable parameters. The effect of varying these parameters on the calculated kinetics and power dependence curves is discussed. A procedure for determining a limited range of threshold dissociation energy, E(o), which fits both the measured induction period and power dependence curves, is presented. Using this procedure, E(o) of leucine enkephalin is determined to be 1.12-1.46 eV. This result is consistent with, although less precise than, values measured previously using blackbody infrared radiative dissociation. Although the blackbody dissociation results were used as a starting point to search for fits of the master equation model to experiment, these results demonstrate that it is, in principle, possible to determine a limited range of E(o) from slow infrared multiphoton dissociation data alone. PMID- 16467894 TI - Databases of Discovery. PMID- 16467899 TI - Structure of cationized glycine, gly.m (m = be, mg, ca, sr, ba), in the gas phase: intrinsic effect of cation size on zwitterion stability. AB - Interactions between divalent metal ions and biomolecules are common both in solution and in the gas phase. Here, the intrinsic effect of divalent alkaline earth metal ions (Be, Mg, Ca, Sr, Ba) on the structure of glycine in the absence of solvent is examined. Results from both density functional and Moller-Plesset theories indicate that for all metal ions except beryllium, the salt-bridge form of the ion, in which glycine is a zwitterion, is between 5 and 12 kcal/mol more stable than the charge-solvated structure in which glycine is in its neutral form. For beryllium, the charge-solvated structure is 5-8 kcal/mol more stable than the salt-bridge structure. Thus, there is a dramatic change in the structure of glycine with increased metal cation size. Using a Hartree-Fock-based partitioning method, the interaction between the metal ion and glycine is separated into electrostatic, charge transfer and deformation components. The charge transfer interactions are more important for stabilizing the charge solvated structure of glycine with beryllium relative to magnesium. In contrast, the difference in stability between the charge-solvated and salt-bridge structure for magnesium is mostly due to electrostatic interactions that favor formation of the salt-bridge structure. These results indicate that divalent metal ions dramatically influence the structure of this simplest amino acid in the gas phase. PMID- 16467897 TI - Redox Regulation of Apoptosis before and after Cytochrome C Release. AB - Programmed cell death, or apoptosis, is one of the most studied areas of modern biology. Apoptosis is a genetically regulated process, which plays an essential role in the development and homeostasis of higher organisms. Mitochondria, known to play a central role in regulating cellular metabolism, was found to be critical for regulating apoptosis induced under both physiological and pathological conditions. Mitochondria are a major source of reactive oxygen species (ROS) but they can also serve as its target during the apoptosis process. Release of apoptogenic factors from mitochondria, the best known of which is cytochrome c, leads to assembly of a large apoptosis-inducing complex called the apoptosome. Cysteine proteases (called caspases) are recruited to this complex and, following their activation by proteolytic cleavage, activate other caspases, which in turn target for specific cleavage a large number of cellular proteins. The redox regulation of apoptosis during and after cytochrome c release is an area of intense investigation. This review summarizes what is known about the biological role of ROS and its targets in apoptosis with an emphasis on its intricate connections to mitochondria and the basic components of cell death. PMID- 16467902 TI - Untangling the Links among Athletic Involvement, Gender, Race, and Adolescent Academic Outcomes. AB - Although previous research has established that high school sports participation may be associated with positive academic outcomes, the parameters of the relationship remain unclear. Using a longitudinal sample of nearly 600 Western New York adolescents, this study examined gender- and race-specific differences in the impact of two dimensions of adolescent athletic involvement ("jock" identity and athlete status) on changes in school grades and school misconduct over a two-year interval. Female and black adolescents who identified themselves as "jocks" reported lower grades than those who did not, whereas female athletes reported higher grades than female nonathletes. Jocks also reported significantly more misconduct (including skipping school, cutting classes, having someone from home called to the school for disciplinary purposes, and being sent to the principal's office) than nonjocks. Gender moderated the relationship between athlete status and school misconduct; athletic participation had a less salutary effect on misconduct for girls than for boys. PMID- 16467900 TI - An Exploratory Investigation of the Association between Clinicians' Attitudes toward Twelve-step Groups and Referral Rates. AB - Affiliation with 12-step groups has been consistently linked to the achievement of abstinence among persons experiencing alcohol and other drug problems. Clinicians play a critical role in fostering clients' engagement in 12-step, yet, little is known about clinicians' attitudes and beliefs about 12-step groups, or about the association between such beliefs and referral practices. This exploratory study investigates this association to gain a greater understanding of determinants of referral practices. Participants were 100 clinicians working within outpatient treatment programs in New York City. Participants held highly positive views of 12-step groups in terms of helpfulness to recovery, but a large percentage endorsed items describing potential points of resistance to 12-step groups, in particular the emphasis such groups place on spirituality and powerlessness. More positive attitudes were associated with greater rates of referral, while resistance to the concepts of spirituality/powerlessness was associated with lower rates of referral. Implications of findings for clinical settings are discussed as well as a research agenda designed to more fully elucidate determinants of clinicians' 12-step referrals. PMID- 16467903 TI - Signaling of the Human P2Y(1) Receptor Measured by a Yeast Growth Assay with Comparisons to Assays of Phospholipase C and Calcium Mobilization in 1321N1 Human Astrocytoma Cells. AB - The human P2Y(1) receptor was expressed in the yeast Saccharomyces cerevisiae strain MPY578q5, which is engineered to couple to mammalian G protein-coupled receptors (GPCRs) and requires agonist-induced activation for growth. A range of known P2Y(1) receptor agonists were examined with the yeast growth assay system, and the results were validated by comparing with potencies in the transfected 1321N1 astrocytoma cell line, in which calcium mobilization was measured with a FLIPR (fluorescence-imaging plate reader). The data were also compared with those from phospholipase C activation and radioligand binding with the use of a newly available radioligand [H]MRS2279 (2-chloro- N-methyl-(N)-methanocarba-2' deoxyadenosine-3',5'-bisphosphate). In the yeast growth assay, the rank order of potency of 2-MeSADP (2-methylthioadenosine 5'-diphosphate), ADP (adenosine 5' diphosphate), and ATP (adenosine 5'-triphosphate) is the same as those in other assay systems, i.e., 2-MeSADP>ADP>ATP. The P2Y(1)-selective antagonist MRS2179 (N methyl-2-deoxyadenosine-3',5'-bisphosphate) was shown to act as an antagonist with similar potency in all systems. The results suggest that the yeast expression system is suitable for screening P2Y(1) receptor ligands, both agonists and antagonists. The yeast system should be useful for random mutagenesis of GPCRs to identify mutants with certain properties, such as selective potency enhancement for small synthetic molecules and constitutive activity. PMID- 16467904 TI - A highly discriminatory multilocus microsatellite typing (MLMT) system for Penicillium marneffei. AB - For eukaryotic pathogens that are depauperate in genetic variation, multilocus microsatellite typing (MLMT) offers an accurate and reproducible method of characterizing genetic diversity; herein we describe the development of an MLMT system for the emerging pathogenic fungus Penicillium marneffei based on 23 microsatellite loci. Screening isolates held within the Centraalbureau voor Schimmelcultures culture collection demonstrate high levels of genetic diversity and 100% reproducibility. This MLMT system provides a powerful epidemiological tool to analyse the underlying parameters that are responsible for the emergence of P. marneffei in human HIV-positive populations. PMID- 16467905 TI - Early Intervention Practices for Children With Autism: Descriptions From Community Providers. AB - Across the country, states are reporting increases in the number of children with autism enrolled in the education system. Although a few specific treatment methods have been established as efficacious for some children with autism in controlled settings, research examining the translation of these treatments into early intervention programs has been minimal. The current study examined provider self-reports of the use of interventions in community settings through focus groups. Providers report the use of both evidence-based and non-evidence-based techniques and indicate that they often combine and modify these techniques based on child, personal, and external factors. Few providers had a clear understanding of evidence-based practice, and all providers reported concerns about adequate training. Implications for early intervention research are discussed. PMID- 16467906 TI - The body schema and the multisensory representation(s) of peripersonal space. AB - In order to guide the movement of the body through space, the brain must constantly monitor the position and movement of the body in relation to nearby objects. The effective 'piloting' of the body to avoid or manipulate objects in pursuit of behavioural goals (Popper & Eccles, 1977, p. 129), requires an integrated neural representation of the body (the 'body schema') and of the space around the body ('peripersonal space'). In the review that follows, we describe and evaluate recent results from neurophysiology, neuropsychology, and psychophysics in both human and non-human primates that support the existence of an integrated representation of visual, somatosensory, and auditory peripersonal space. Such a representation involves primarily visual, somatosensory, and proprioceptive modalities, operates in body part-centred reference frames, and demonstrates significant plasticity. Recent research shows that the use of tools, the viewing of one's body or body parts in mirrors, and in video-monitors, may also modulate the visuotactile representation of peripersonal space. PMID- 16467907 TI - A psychological approach to understanding the social and language impairments in autism. AB - This paper surveys current research on the social and communicative impairments in autism. In diagnostic schemes, the criteria for identifying autism in these domains include overlapping features. One approach to interpreting this overlap is to consider that social and communicative impairments reflect the same underlying cognitive deficit, referred to as the 'theory of mind' hypothesis of autism. On this view autism involves primary difficulties in identifying mental states in other people, and in interpreting behavior and action in relation to a person's mental state. Studies on the relationship between social behavior, communicative functioning, and theory of mind in children with autism are reviewed, emphasizing the connections between these areas of impairment that are central to the definition of the autistic syndrome. PMID- 16467908 TI - The influence of language on theory of mind: a training study. AB - This study investigated the role of language in the development of theory of mind. It was hypothesized that the acquisition of the syntactic and semantic properties of sentential complements would facilitate the development of a representational theory of mind. Sixty preschoolers who failed false belief and sentential complement pretests were randomly assigned to training on false belief, sentential complements, or relative clauses (as a control group). All the children were post-tested on a set of different theory of mind tasks, sentential complements and relative clauses. The main findings were that the group trained on sentential complements not only acquired the linguistic knowledge fostered by the training, but also significantly increased their scores on a range of theory of mind tasks. In contrast, false belief training only led to improved theory of mind scores but had no influence on language. The control group, trained on relative clauses, showed no improvement on theory of mind posttests. These findings are taken as evidence that the acquisition of sentential complements contributes to the development of theory of mind in preschoolers. PMID- 16467910 TI - Prediction of Feeding Performance in Preterm Infants. AB - A nonexperimental study with a sample of 95 preterm infants was used to develop a model of feeding performance outcomes (proficiency, percent of prescribed volume consumed, and efficiency) using feeding readiness indicators (morbidity, maturity, behavior state at feeding start, and feeding experience). All readiness indicators were related to each other. In particular, there was a strong relationship between maturity and feeding experience. Morbidity only had an effect on efficiency; the most ill infants were less efficient feeders. Behavior state affects all feeding outcomes in a linear fashion; more awake and alert infants had better feeding performance outcomes. The effect of experience and maturity on the outcomes is more complex because of the relationship between the two. Experience has a greater effect as the infant matures, and as the infant matures, the amount of experience increases. Successful feeding requires maturity as well as experience. PMID- 16467911 TI - Modeling the maximum charge state of arginine-containing Peptide ions formed by electrospray ionization. AB - A model for the gas-phase proton transfer reactivity of multiply protonated molecules is used to quantitatively account for the maximum charge states of a series of arginine-containing peptide ions measured by Downard and Biemann (Int. J. Mass Spectrom. Ion Processes 1995, 148, 191-202). We find that our calculations account exactly for the maximum charge state for 7 of the 10 peptides and are off by one charge for the remaining 3. These calculations clearly predict the trend in maximum charge states for these peptides and provide further evidence that the maximum charge state of ions formed by electrospray ionization is determined by their gas-phase proton transfer reactivity. PMID- 16467912 TI - What Can We Learn From Longitudinal Studies of Adult Development? AB - This article distinguishes between normal and pathological aging, provides an interdisciplinary context, and then considers a sample case of cognitive aging. Developmental influences on cognition include the physiological infrastructure, genetic predispositions, and environmental influences. Different types of longitudinal studies are distinguished, and contrasting findings of cross sectional and longitudinal are examined in the sample case of the Seattle Longitudinal Study. Also considered is the longitudinal context for intervention studies and the role of longitudinal family studies in assessing rate of aging and generational differences in rates of aging. Finally, attention is given to the role of longitudinal studies in the early detection of risk for dementia in advanced age. PMID- 16467914 TI - Telos: The Revival of an Aristotelian Concept in Present Day Ethics. AB - Genetic engineering is often looked upon with disfavour on the grounds that it involves 'tampering with nature'. Most philosophers do not take this notion seriously. However, some do. Those who do tend to understand nature in an Aristotelian sense, as the essence or form which is the final end or telos for the sake of which individual organisms live, and which also explains why they are as they are. But is this really a tenable idea? In order to secure its usage in present day ethics, I will first analyze the contexts in which it is applied today, then discuss the notion of telos as it was employed by Aristotle himself, and finally debate its merits and defend it, as far as possible, against common objections. PMID- 16467913 TI - Nanomedicine and protein misfolding diseases. AB - Misfolding and self assembly of proteins in nano-aggregates of different sizes and morphologies (nano-ensembles, primarily nanofilaments and nano-rings) is a complex phenomenon that can be facilitated, impeded, or prevented, by interactions with various intracellular metabolites, intracellular nanomachines controlling protein folding and interactions with other proteins. A fundamental understanding of molecular processes leading to misfolding and self-aggregation of proteins involved in various neurodegenerative diseases will provide critical information to help identify appropriate therapeutic routes to control these processes. An elevated propensity of misfolded protein conformation in solution to aggregate with the formation of various morphologies impedes the use of traditional physical chemical approaches for studies of misfolded conformations of proteins. In our recent alternative approach, the protein molecules were tethered to surfaces to prevent aggregation and AFM force spectroscopy was used to probe the interaction between protein molecules depending on their conformations. It was shown that formation of filamentous aggregates is facilitated at pH values corresponding to the maximum of rupture forces. In this paper, a novel surface chemistry was developed for anchoring of amyloid beta (Abeta) peptides at their N-terminal moieties. The use of the site specific immobilization procedure allowed to measure the rupture of Abeta-Abeta contacts at single molecule level. The rupture of these contacts is accompanied by the extension of the peptide chain detected by a characteristic elasto-mechanical component of the force-distance curves. Potential applications of the nanomechanical studies to understanding the mechanisms of development of protein misfolding diseases are discussed. PMID- 16467916 TI - Glioma cell integrin expression and their interactions with integrin antagonists: Research Article. AB - A panel of human glioma cell explants was screened for integrin expression by flow cytometry using alpha(nu)beta-specific antibodies. A lower percentage of the glioma cells were positive for the alpha(nu)beta3 (mean % positive = 20.8%) integrin, whereas higher percentages were positive for the alphanubeta5 (mean % positive = 72.7%), VLA5alpha (mean % positive = 87%) and VLAbeta1 (mean % positive = 41.7%) integrins. A series of RGD peptides was designed, synthesized and tested for binding to integrin receptors. Based on the results of the binding to the isolated integrin receptors and the expression of integrins on glioma cell lines, a peptide that binds potently to the alpha(nu)beta3, alpha(nu)beta5 and alpha(5)beta(1) was selected for further investigations with regards to its effect on glioma cells. The peptide, Ac-c[(Pen)-Tyr(Me)-Ala-Arg-Gly-Asp-Asn-Tic Cys]NH(2) (RGD peptide), exhibited high potential for use in clinical intracranial administration since it had good stability in rat brain cell homogenates placed into artificial cerebrospinal fluid. Using an HPLC method for quantification of peptides in rat brain cell homogenates, we could demonstrate the half-life of the RGD peptide approximated 20 hr. Relative to a scrambled peptide control (non-RGD sequence, same amino acids), the experimental RGD peptide significantly decreased glioma cell proliferation of the entire panel of rat and human glioma cells tested. Adhesion of recently passaged glioma cells to glioma-derived extracellular matrix protein-coated plates was inhibited significantly by the RGD peptide. The peptide also reversed attachment of plated glioma cells. The RGD peptide caused some, but not substantial, glioma cell injury, as evidenced by a quantitative in vitro nuclear DNA morphologic assay and by a flow cytometric assay employing 7-amino actinomycin D (7AAD). We histologically monitored for toxicity caused by various doses of the RGD peptide infused repeatedly into normal cannulated rat brain. At safe doses, the experimental RGD peptide-treated brains did not show significant differences from those infused with scrambled peptide or buffer-treated controls. In tumor-bearing brains, slightly smaller tumor areas were measured with a higher necrotic-to tumor index in the RGD peptide treated relative to the scrambled peptide-treated controls. This was obtained with intracranial peptide administrations or combined intracranial and intraperitoneal injections. From this in vitro work, we conclude that the anti-glioma effects of the RGD peptide tested resulted from lowered glioma proliferation and adhesion/mobility, rather than from significant glioma cell injury in the timeframe analyzed. Although other mechanisms not discerned from our limited histopathological observations may be operational, from our in vivo work, we conclude that repeated administration of RGD peptide into brain is safe but that better delivery of the peptides to infiltrating tumor cells is necessary. PMID- 16467915 TI - Catalytic nucleic acid enzymes for the study and development of therapies in the central nervous system: Review Article. AB - Nucleic acid enzymes have been used with great success for studying natural processes in the central nervous system (CNS). We first provide information on the structural and enzymatic differences of various ribozymes and DNAzymes. We then discuss how they have been used to explore new therapeutic approaches for treating diseases of the CNS. They have been tested in various systems modeling retinitis pigmentosum, proliferative vitreoretinopathy, Alzheimer's disease, and malignant brain tumors. For these models, effective targets for nucleic acid enzymes have been readily identified and the rules for selecting cleavage sites have been well established. The bulk of studies, including those from our laboratory, have emphasized their use for gliomas. With the availability of multiple excellent animal models to test glioma treatments, good progress has been made in the initial testing of nucleic acid enzymes for brain tumor therapy. However, opportunities still exist to significantly improve the delivery and efficacy of ribozymes to achieve effective treatment. The future holds significant potential for the molecular targeting and therapy of eye diseases, neurodegenerative disorders, and brain tumors with these unique treatment agents. PMID- 16467917 TI - Neuropsychological frameworks for understanding autism. AB - Neuropsychological theories have traditionally attempted to provide a unifying account of the complex and diverse behavioral manifestations of autism in terms of their underlying psychological mechanisms and associated brain bases. This article reviews three competing neuropsychological theories of autism: the executive dysfunction hypothesis, the weak central coherence hypothesis, and the limbic system hypothesis. Each theory is evaluated critically with regard to the primary neuropsychological deficit hypothesized and the research findings that have been offered in support of it. In a concluding section, some of the metatheoretical assumptions informing attempts to identify a "core" neuropsychological impairment in autism are outlined and questioned, and new approaches to a neuropsychological understanding of autism are suggested. PMID- 16467920 TI - Circuits of Surveillance. AB - This paper examines the increasing police use of DNA profiling and databasing as a developing instrumentality of modern state surveillance. It briefly notes previously published work on a variety of surveillance technologies and their role in the governance of social action and social order. It then argues that there are important differences amongst the ways in which several such technologies construct and use identificatory artefacts, their orientations to human subjectivity, and their role in the governmentality of citizens and others. The paper then describes the novel and powerful form of bio-surveillance offered by DNA profiling and illustrates this by reference to an ongoing empirical study of the police uses of the UK National DNA Database for the investigation of crime. It is argued that DNA profiling and databasing enable the construction of a 'closed circuit' of surveillance of a defined population. PMID- 16467919 TI - Isomerization/Recyclization of some 5-Ethoxycarbonyl-pyrimidines. AB - This communication reports on the investigation of a new recyclization conversion of a pyrimidine ring, which can be referred to as C-C recyclization. In this reaction the nucleophile cleaves the pyrimidine ring at the N(3)-C(4) bond, and following rotation around the single C(5)-C(6) bond the new cyclization takes place. This type of recyclization has general applicability, and takes place upon alkali treatment of substituted 4-methyl-5-ethoxycarbonyl- and 4-amino-5 ethoxycarbonyl-pyrimidines (1) which are transformed respectively to 4-hydroxy-5 acetyl- and 4-hydroxy-5-carbamoylpyrimidines (2). The obtained pyrimidyl-ketones can be readily converted to their hydrazones 7-12. PMID- 16467921 TI - Genetics and Forensics: Making the National DNA Database. AB - This paper is based on a current study of the growing police use of the epistemic authority of molecular biology for the identification of criminal suspects in support of crime investigation. It discusses the development of DNA profiling and the establishment and development of the UK National DNA Database (NDNAD) as an instance of the 'scientification of police work' (Ericson and Shearing 1986) in which the police uses of science and technology have a recursive effect on their future development. The NDNAD, owned by the Association of Chief Police Officers of England and Wales, is the first of its kind in the world and currently contains the genetic profiles of more than 2 million people. The paper provides a framework for the examination of this socio-technical innovation, begins to tease out the dense and compact history of the database and accounts for the way in which changes and developments across disparate scientific, governmental and policing contexts, have all contributed to the range of uses to which it is put. PMID- 16467922 TI - Cognitive mediation of childhood maltreatment and adult depression in recent crime victims. AB - Childhood maltreatment has been linked to adult depressive disorders. However, few studies have examined mechanisms through which childhood maltreatment may contribute to adult depression. Thus, we examined the role of one potential mechanism of this relationship, maladaptive cognitions, in a recently traumatized sample. Participants were adult women who had been recently raped (n = 133) or physically assaulted (n = 73). We examined whether maladaptive self-and other cognitions mediated relationships between childhood sexual, physical, and emotional abuse and current depression. Relationships between childhood sexual abuse and both current depression symptoms and diagnosis were mediated by maladaptive cognitions about self. Relationships between both childhood sexual abuse and childhood physical abuse and adult depressive symptoms were mediated by maladaptive cognitions about others. PMID- 16467923 TI - Surface functionalization of gold nanoparticles using hetero-bifunctional poly(ethylene glycol) spacer for intracellular tracking and delivery. AB - For the development of surface-functionalized gold nanoparticles as cellular probes and delivery agents, we have synthesized hetero-bifunctional poly(ethylene glycol) (PEG, MW 1500) having a thiol group on one terminus and a reactive functional group on the other for use as a flexible spacer. Coumarin, a model fluorescent dye, was conjugated to one end of the PEG spacer and gold nanoparticles were modified with coumarin-PEG-thiol. Surface attachment of coumarin through the PEG spacer decreased the fluorescence quenching effect of gold nanoparticles. The results of cellular cytotoxicity and fluorescence confocal analyses showed that the PEG spacer-modified nanoparticles were essentially non-toxic and could be efficiently internalized in the cells within 1 hour of incubation. Intracellular particle tracking using a Keck 3-D Fusion Microscope System showed that the functionalized gold nanoparticles were rapidly internalized in the cells and localized in the peri-nuclear region. Using the PEG spacer, the gold nano-platform can be conjugated with a variety of biologically relevant ligands such as fluorescent dyes, antibodies, etc in order to target, probe, and induce a stimulus at the target site. PMID- 16467924 TI - Electronic Structure of the Aqueous Vanadyl Ion Probed by 9 and 94 GHz EPR and Pulsed ENDOR Spectroscopies and Density Functional Theory Calculations. AB - The aqueous vanadyl ion ([VO(H(2)O)(5)](2+)) has been investigated by X-band EPR, 94 GHz W-band EPR, and ESE-ENDOR. These experiments reveal information about the hyperfine (|A(xx)| = 208.5 MHz, |A(yy)| = 208.5 MHz, |A(zz)| = 547.0 MHz), and nuclear quadrupole coupling (|e(2)qQ| = 5.6 MHz) of the (51)V nucleus. The measured nuclear quadrupole coupling parameters are compared to values determined by density functional theory calculations (|e(2)qQ| = 5.2 MHz). These theoretical calculations illustrate that axial ligands and molecular distortions can alter the magnitude of the nuclear quadrupole interaction. PMID- 16467926 TI - Fabrication of nanoelectrodes for neurophysiology: cathodic electrophoretic paint insulation and focused ion beam milling. AB - The fabrication and characterization of tungsten nanoelectrodes insulated with cathodic electrophoretic paint is described together with their application within the field of neurophysiology. The tip of a 127 mum diameter tungsten wire was etched down to less than 100 nm and then insulated with cathodic electrophoretic paint. Focused ion beam (FIB) polishing was employed to remove the insulation at the electrode's apex, leaving a nanoscale sized conductive tip of 100-1000 nm. The nanoelectrodes were examined by scanning electron microscopy (SEM) and their electrochemical properties characterized by steady state linear sweep voltammetry. Electrode impedance at 1 kHz was measured too. The ability of a 700 nm tipped electrode to record well-isolated action potentials extracellularly from single visual neurons in vivo was demonstrated. Such electrodes have the potential to open new populations of neurons to study. PMID- 16467927 TI - Control Issues in High-speed AFM for Biological Applications: Collagen Imaging Example. AB - This article considers the precision positioning problem associated with high speed operation of the Atomic Force Microscope (AFM), and presents an inversion based control approach to achieve precision positioning. Although AFMs have high (nanoscale) spatial resolution, a problem with current AFM systems is that they have low temporal resolution, i.e., AFM imaging is slow. In particular, current AFM imaging cannot be used to provide three-dimensional, time-lapse images of fast processes when imaging relatively-large, soft samples. For instance, current AFM imaging of living cells takes 1-2 minutes (per image frame) - such imaging speeds are too slow to study rapid biological processes that occur in seconds, e.g., to investigate the rapid movement of cells or the fast dehydration and denaturation of collagen. This inability, to rapidly image fast biological processes, motivates our current research to increase the operating speed of the AFM. We apply an inversion-based feedback/feedforward control approach to overcome positioning problems that limit the operating speed of current AFM systems. The efficacy of the method, to achieve high-speed AFM operation, is experimentally evaluated by applying it to image collagen samples. PMID- 16467929 TI - Unimolecular reaction kinetics in the high-pressure limit without collisions. AB - Molecular activation by blackbody photons, first postulated in 1919 by Perrin, plays a dominant role in the unimolecular dissociation of large ions trapped at low pressure in a Fourier-transform mass spectrometer. Under readily achievable experimental conditions, molecular ions of the protein ubiquitin equilibrate with the blackbody radiation field inside the vacuum chamber. The internal energy of a population of these ions is given by a Boltzmann distribution. From the temperature dependence of unimolecular dissociation rate constants measured in the zero-pressure limit, Arrhenius activation parameters equal to those in the high-pressure limit are obtained. PMID- 16467928 TI - Neuromusculoskeletal modeling: estimation of muscle forces and joint moments and movements from measurements of neural command. AB - This paper provides an overview of forward dynamic neuromusculoskeletal modeling. The aim of such models is to estimate or predict muscle forces, joint moments, and/or joint kinematics from neural signals. This is a four-step process. In the first step, muscle activation dynamics govern the transformation from the neural signal to a measure of muscle activation-a time varying parameter between 0 and 1. In the second step, muscle contraction dynamics characterize how muscle activations are transformed into muscle forces. The third step requires a model of the musculoskeletal geometry to transform muscle forces to joint moments. Finally, the equations of motion allow joint moments to be transformed into joint movements. Each step involves complex nonlinear relationships. The focus of this paper is on the details involved in the first two steps, since these are the most challenging to the biomechanician. The global process is then explained through applications to the study of predicting isometric elbow moments and dynamic knee kinetics. PMID- 16467930 TI - The Development of Anticipatory Smiling. AB - When do infants begin to communicate positive affect about physical objects to their social partners? We examined developmental changes in the timing of smiles during episodes of initiating joint attention that involved an infant gazing between an object and a social partner. Twenty-six typically developing infants were observed at 8, 10, and 12 months during the Early Social-Communication Scales, a semistructured assessment for eliciting initiating joint attention and related behaviors. The proportion of infant smiling during initiating joint attention episodes did not change with age, but there was a change in the timing of the smiles. The likelihood of infants smiling at an object and then gazing at the experimenter while smiling (anticipatory smiling) increased between 8 and 10 months and remained stable between 10 and 12 months. The increase in the number of infants who smiled at an object and then made eye contact suggests a developing ability to communicate positive affect about an object. PMID- 16467931 TI - Supported metalloporphyrin catalysts for alkene epoxidation. AB - This review is devoted to the recent advances in the preparation of immobilised metalloporphyrins and their use as heterogeneous catalysts for alkene epoxidation. The wide range of supports, nature of attachments, and metalloporphyrins that have been reported is detailed and a comparison is made between the activities of the resulting catalysts in the epoxidation of different alkenes. The important issue of recyclability of the metalloporphyrins is also covered. PMID- 16467932 TI - Lectin recognition of a new SOD mimic bioconjugate studied with surface plasmon resonance imaging. AB - Surface plasmon resonance imaging is used to demonstrate the recognition by the Ricinus communis agglutinin of a new SOD mimic, a bioconjugate of the manganese(II) complex of 1,4,7,10,13-pentaazacyclopentadecane with galactose. PMID- 16467933 TI - PegPhos: a monodentate phosphoramidite ligand for enantioselective rhodium catalysed hydrogenation in water. AB - A BICOL derived monodentate phosphoramidite ligand gives ee's up to 89% in the enantioselective Rh-catalysed hydrogenation of N-acyl dehydroalanine using water as the solvent. PMID- 16467934 TI - Archaeal C80 isoprenoid tetraacids responsible for naphthenate deposition in crude oil processing. AB - The structure of a novel class of octaterpene tetracarboxylic acids which is responsible for naphthenate deposition in crude oil processing has been determined by NMR and mass spectroscopy. PMID- 16467935 TI - Delineation of the factors governing reactivity and selectivity in epoxide formation from ammonium ylides and aldehydes. AB - Diastereoselectivity in reactions of aryl-stabilised ammonium ylides are highly sensitive to the nature of the amine and the ylide substituent. DFT calculations are consistent with a mechanism in which reversibility in betaine formation [despite the high energy (and therefore instability) of ammonium ylides] is finely balanced due to the high barrier to ring closure. PMID- 16467936 TI - A unique NH-spacer for N-benzamidothiourea based anion sensors. Substituent effect on anion sensing of the ICT dual fluorescent N-(p-dimethylaminobenzamido) N'-arylthioureas. AB - A series of N-(p-dimethylaminobenzamido)-N'-(substituted-phenyl)thioureas (substituent = p-CH3, H, p-Cl, p-Br, m-Br, m-NO2, and p-NO2) were designed as anion sensors in order to better understand the -NH-spacer via a substituent effect investigation. In these molecules the dual fluorescent intramolecular charge transfer (ICT) fluorophore p-dimethylaminobenzamide as the signal reporter was linked to the anion-binding site, the thiourea moiety, via an N-N single bond. Correlation of the NMR signals of the aromatic and -NH protons with substituents in these molecules indicated that the N-N single bond stopped the ground-state electronic communication between the signal reporter and the anion binding site. Dual fluorescence was observed in highly polar solvents such as acetonitrile with the former five derivatives. The fact that the CT emission wavelength and the CT to LE emission intensity ratio of the sensors were independent of the substituent existing in the anion-binding moiety suggested that the substituent electronic effect could not be communicated to the CT fluorophore in the excited-state either. Yet in acetonitrile both the CT dual fluorescence and the absorption of the sensors were found to be highly sensitive toward anions. A conformation change around the N-N bond in the sensor molecules was suggested to occur upon anion binding that established the electronic communication between the signal reporter and the anion-binding site. The anion binding constants of the N-(p-dimethylaminobenzamido)thiourea sensors were found higher than those of the corresponding traditional N-phenylthiourea counterparts and the substituent effect on the anion binding constant was much higher than that in the latter. "-NH-" was shown to be a unique spacer that affords N benzamidothiourea allosteric anion sensors. PMID- 16467937 TI - Spontaneity in the patellamide biosynthetic pathway. AB - Post-translationally modified ribosomal peptides are unusual natural products and many have potent biological activity. The biosynthetic processes involved in their formation have been delineated for some, but the patellamides represent a unique group of these metabolites with a combination of a macrocycle, small heterocycles and d-stereocentres. The genes encoding for the patellamides show very low homology to known biosynthetic genes and there appear to be no explicit genes for the macrocyclisation and epimerisation steps. Using a combination of literature data and large-scale molecular dynamics calculations with explicit solvent, we propose that the macrocyclisation and epimerisation steps are spontaneous and interdependent and a feature of the structure of the linear peptide. Our study suggests the steps in the biosynthetic route are heterocyclisation, macrocyclisation, followed by epimerisation and finally dehydrogenation. This study is presented as testable hypothesis based on literature and theoretical data to be verified by future detailed experimental investigations. PMID- 16467938 TI - RNA binding and thiolytic stability of a quinoline-containing helix-threading peptide. AB - Helix-threading peptides (HTPs) bind selectively to sites predisposed to intercalation in folded RNA molecules placing peptide functional groups into the dissimilar grooves of the duplex. Here we report the design and synthesis of new HTPs with quinoline as the intercalation domain. A quinoline-containing HTP is shown to bind selectively to duplex RNA binding sites. Furthermore, the affinity cleavage pattern generated using an EDTA.Fe modified derivative is consistent with minor groove localization of its N-terminus. This compound binds base-pair steps flanked by single nucleotide bulges on the 3' side on both strands, whereas bulges on the 5' side of the intercalation site do not support binding. Furthermore, unlike acridine HTPs, the quinoline compound is resistant to thiolytic degradation that leads to loss of RNA-binding activity. The RNA-binding selectivity and stability observed for quinoline-containing HTPs make them excellent candidates for further development as regulators of intracellular RNA function. PMID- 16467939 TI - A computational study on the amine-oxidation mechanism of monoamine oxidase: insight into the polar nucleophilic mechanism. AB - The proposed polar nucleophilic mechanism of MAO was investigated using quantum chemical calculations employing the semi-empirical PM3 method. In order to mimic the reaction at the enzyme's active site, the reactions between the flavin and the p-substituted benzylamine substrate analogs were modeled. Activation energies and rate constants of all the reactions were calculated and compared with the published experimental data. The results showed that electron-withdrawing groups at the para position of benzylamine increase the reaction rate. A good correlation between the log of the calculated rate constants and the electronic parameter (sigma) of the substituent was obtained. These results agree with the previous kinetic experiments on the effect of p-substituents on the reduction of MAO-A by benzylamine analogs. In addition, the calculated rate constants showed a correlation with the rate of reduction of the flavin in MAO-A. In order to verify the results obtained from the PM3 method single-point B3LYP/6-31G*//PM3 calculations were performed. These results demonstrated a strong reduction in the activation energy for the reaction of benzylamine derivatives having electron withdrawing substituents, which is in agreement with the PM3 calculations and the previous experimental QSAR study. PM3 and B3LYP/6-31G* energy surfaces were obtained for the overall reaction of benzylamine with flavin. Results suggest that PM3 is a reasonable method for studying this kind of reaction. These theoretical findings support the proposed polar nucleophilic mechanism for MAO-A. PMID- 16467940 TI - Utilization of 3'-carboxy-containing tyrosine derivatives as a new class of phosphotyrosyl mimetics in the preparation of novel non-phosphorylated cyclic peptide inhibitors of the Grb2-SH2 domain. AB - A new class of phosphotyrosyl (pTyr) mimetics, distinct from the conventional pTyr mimetic design of adding non-hydrolyzable acidic functionalities to the 4' position of phenylalanine, was created by introducing carboxy-containing groups to the 3'-position of tyrosine. The effect of the chain length of the carboxy substituent was examined. Reported herein is the chiral pool synthesis of the new pTyr mimetics, and their first use in a novel non-phosphorylated Grb2-SH2 domain binding motif with the 5-amino-acid sequence Xx1-Leu-(3'-substituted-Tyr)-Ac6c Asn. The highest affinity was exhibited by the 3-L-(2-carboxyethyl)tyrosine containing sulfoxide-cyclized peptide , with an IC50 = 1.1 microM, providing a promising new template for further development of potent Grb2-SH2 domain inhibitors with reduced charge and peptidic nature, but improved selectivity and bioavailability. PMID- 16467941 TI - A versatile template for the formation of [2]pseudorotaxanes. 1,2 Bis(pyridinium)ethane axles and 24-crown-8 ether wheels. AB - Linear 1,2-bis(pyridinium)ethane 'axles' and macrocyclic 24-membered crown ether 'wheels' (, and ) combine to form [2]pseudorotaxanes. These interpenetrated adducts are held together by N+...O ion-dipole interactions, a series of C-H...O hydrogen bonds and pi-stacking between electron-poor pyridinium rings of the axle and electron-rich catechol rings of the wheel. 1H NMR spectroscopy was used to identify the structural details of the interaction and to determine the thermodynamics of the binding process in solution. Analysis of nine of these adducts by single crystal X-ray crystallography allowed a detailed study of the non-covalent interactions in the solid state. A wide variety of structural changes could be made to the system. The versatility and potential of the template for the construction of permanently interlocked structures such as rotaxanes and catenanes is discussed. PMID- 16467942 TI - Synthesis of bisubstrate analogues targeting alpha-1,3-fucosyltransferase and their activities. AB - We designed two bisubstrate analogues targeting alpha-1,3-fucosyltransferases, based on the three dimensional structure of Lewis X, which is the product of a alpha-1,3-fucosyltransferase reaction. We selected guanosine-5'-diphospho-L galactose as a donor mimic and 2-hydroxyethyl beta-D-galactoside as an acceptor mimic, and tethered these two mimics with a methylene or ethylene linker. For the synthesis, the 6-position of L-galactose and the 6-position of D-galactose were first tethered via a methylene or ethylene linker. The L-galactose moiety was then converted to a GDP derivative. Both bisubstrate analogues were moderate inhibitors against alpha-1,3-fucosyltransferase V and VI. The fact that they were substrates of alpha-1,3-fucosyltransferase VI suggested that these compounds bound to the donor binding site, but not to the acceptor binding site. PMID- 16467943 TI - Unprecedented chemical structure and biomimetic synthesis of erucalexin, a phytoalexin from the wild crucifer Erucastrum gallicum. AB - The isolation, structure determination, total synthesis and antifungal activity of erucalexin, a novel phytoalexin produced by the wild crucifer dog mustard are described. Erucalexin is a structurally unique plant alkaloid, representing the first example of a spiro[2H-indole-2,5'(4'H)-thiazol]-3-one, likely derived from a C-3-C-2 carbon migration in a 3-substituted indolyl nucleus. PMID- 16467945 TI - The influence of phenyl and phenoxy modification in the hydrophobic tails of di-n alkyl phosphate amphiphiles on aggregate morphology. AB - A series of di-n-alkyl phosphate amphiphiles containing phenyl and phenoxy groups in the hydrophobic tails were synthesised, and their aggregation behaviour was investigated using fluorescence spectroscopy, differential scanning calorimetry, and cryo-electron microscopy. The aggregates displayed a wide variety of aggregate morphologies. The incorporation of a phenyl group into the end or in the middle of the alkyl chain lowered the main phase transition temperature, resulting in closed vesicles only above the phase transition temperature. Introducing a phenoxy group at the end of the alkyl chain resulted in open bilayer structures and bicelles. PMID- 16467944 TI - Non-equivalence of D- and L-trehalose in stabilising alkaline phosphatase against freeze-drying and thermal stress. Is chiral recognition involved? AB - Comparison of the ability of the enantiomeric forms of trehalose to stabilise alkaline phosphatase towards dehydration and heat showed that natural D-trehalose is superior to L-trehalose, although both disaccharides provide some protection for the enzyme. The result of this novel experiment suggests a chiral differentiation between carbohydrate and protein and thus lends support for the water replacement hypothesis of solute-based stabilisation of biomolecules, but the non-crystallinity and the physical form of the L-isomer may also be a key factor. PMID- 16467946 TI - Formation of lactones from sialylated MUC1 glycopeptides. AB - The tumor-associated carbohydrate antigens TN, T, sialyl TN and sialyl T are expressed on mucins in several epithelial cancers. This has stimulated studies directed towards development of glycopeptide-based anticancer vaccines. Formation of intramolecular lactones involving sialic acid residues and suitably positioned hydroxyl groups in neighboring saccharide moieties is known to occur for glycolipids such as gangliosides. It has been suggested that these lactones are more immunogenic and tumor-specific than their native counterparts and that they might find use as cancer vaccines. We have now investigated if lactonization also occurs for the sialyl TN and T antigens of mucins. It was found that the model compound sialyl T benzyl glycoside , and the glycopeptide Ala-Pro-Asp-Thr-Arg-Pro Ala from the tandem repeat of the mucin MUC1, in which Thr stands for the 2,3 sialyl-T antigen, lactonized during treatment with glacial acetic acid. Compound gave the 1''--> 2' lactone as the major product and the corresponding 1''--> 4' lactone as the minor product. For glycopeptide the 1''--> 4' lactone constitued the major product, whereas the 1''--> 2' lactone was the minor one. When lactonized was dissolved in water the 1''--> 4' lactone underwent slow hydrolysis, whereas the 1''--> 2' remained stable even after a 30 days incubation. In contrast the corresponding 2,6-sialyl-TN glycopeptide did not lactonize in glacial acetic acid. PMID- 16467947 TI - 15N CIDNP investigations of the peroxynitric acid nitration of L-tyrosine and of related compounds. AB - Peroxynitric acid (O2NOOH) nitrates L-tyrosine and related compounds at pH 2-5. During reaction with O2(15)NOOH in the probe of a 15N NMR spectrometer, the NMR signals of the nitration products of L-tyrosine, N-acetyl-L-tyrosine, 4 fluorophenol and 4-methoxyphenylacetic acid appear in emission indicating a nitration via free radicals. Nuclear polarizations are built up in radical pairs [15NO2* , PhO*]F or [15NO2* , ArH*+]F formed by diffusive encounters of 15NO2 with phenoxyl-type radicals PhO or with aromatic radical cations ArH*+. Quantitative 15N CIDNP investigations with N-acetyl-L-tyrosine and 4-fluorophenol show that the radical-dependent nitration is the only reaction pathway. During the nitration reaction, the 15N NMR signal of 15NO3- also appears in emission. This is explained by singlet-triplet transitions in radical pairs [15NO2* , 15NO3*]S generated by electron transfer between O2(15)NOOH and H15NO2 formed as a reaction intermediate. During reaction of peroxynitric acid with ascorbic acid, 15N CIDNP is again observed in the 15N NMR signal of 15NO3- showing that ascorbic acid is oxidized by free radicals. In contrast to this, O2(15)NOOH reacts with glutathione and cysteine without the appearance of 15N CIDNP, indicating a direct oxidation without participation of free radicals. PMID- 16467948 TI - Incorporation of deuterium-labelled analogs of isopentenyl diphosphate for the elucidation of the stereochemistry of rubber biosynthesis. AB - A series of six deuterium-labelled analogs of isopententyl diphosphate (IPP) was prepared to investigate the detailed stereochemical course of addition of C5 units during rubber biosynthesis in Hevea brasiliensis and Parthenium argentatum. These analogs were incorporated into the cis-polyisoprene chain by rubber transferase in rubber particles, and the stereochemistry was determined by 2H-NMR analysis of the polymer or of levulinic acid derivatives obtained from its ozonolytic degradation. Results indicate that rubber chain elongation occurs with loss of the pro-S hydrogen of IPP, addition of the allylic diphosphate to the si face of IPP and inversion of stereochemistry at the carbon bearing the diphosphate. PMID- 16467949 TI - [Gaucher disease, Fabry disease and mucopolysaccharidosis type I--how can the rheumatologist recognise these patients?]. AB - The lysosomal storage diseases Gaucher disease, Fabry disease and MPS I are rare inheritable metabolic disorders that are now treatable with enzyme replacement therapy. In order to avoid irreversible complications, an early diagnosis and initiation of therapy is important. Due to the musculoskeletal symptoms associated with these storage diseases, patients are likely to visit a rheumatologist, who should, therefore, be able to recognise and diagnose these rare diseases. On the basis of the causal factors behind Gaucher disease, Fabry disease und MPS I (here Scheie syndrome), key symptoms that the rheumatologist (internist or paediatrician) should be familiar with for the differential diagnosis of these patients will be discussed. In addition, a short introduction to the pathophysiology and data on the prognosis and therapy for these diseases will be presented. PMID- 16467950 TI - Ethnic variation in symptoms and response to risperidone in youths with schizophrenia-spectrum disorders. AB - BACKGROUND: Evaluation of symptom presentation and antipsychotic response based on ethnicity in children and adolescents with schizophrenia is limited. The purpose of this naturalistic, retrospective database study was to compare symptom presentation of children and adolescents of different ethnicities with schizophrenia-spectrum disorders, and response to risperidone. METHOD: African American (n = 38), Caucasian (n = 30), or Hispanic (n = 37) youths started on risperidone were eligible. Child Behavior Checklist (CBCL) total, internalizing, and externalizing scores were evaluated at baseline, 90 days, 1 year, and 2 year intervals. RESULTS: At baseline, Hispanic patients had lower CBCL externalizing scores than African-Americans or Caucasians. African-Americans showed significant differences in CBCL total, internalizing, and externalizing scores at 90 days compared to baseline. Hispanics showed improvement in CBCL internalizing scores over the 2 year period. No significant improvements were observed in Caucasians. African-American patients had significantly lower CBCL total at 90 days compared to Hispanic patients. No significant difference existed in 2 year hospitalization rates between groups. CONCLUSIONS: These findings suggest that ethnicity may play a role in symptom presentation and treatment response to risperidone for children and adolescents with schizophrenia-spectrum disorders. Future studies in children and adolescents are necessary to examine ethnospecific differences in antipsychotic use and treatment response. PMID- 16467951 TI - Psychiatric disorders certified on death certificates in an English population. AB - BACKGROUND: Psychiatric disorders are sometimes certified on death certificates, but seldom selected as the underlying cause of death. The majority of deaths with a certified psychiatric cause are usually omitted from official mortality statistics, which are typically based on the underlying cause alone. AIM: To report on death rates for psychiatric disorders, as certified on death certificates, including all mentioned causes as well as the underlying cause of death. METHOD: Analysis of database including all certified causes of death in 1979-1999, in three time periods defined by coding rule changes. RESULTS: Statistics on the underlying cause of death grossly under-estimated certified psychiatric disorders. For example, in the first period of our study they missed 88% of deaths in which schizophrenia was a certified cause, 98% of affective psychosis, and 96% of depression. Over time, considering all certified causes, age-standardised death rates for schizophrenia declined, those for affective psychosis showed no change, and those for depression and dementia increased. CONCLUSION: The decline in mortality rates for schizophrenia, and the increase for depression and dementia, may reflect real changes over time in disease prevalence at death, although other explanations are possible and are discussed. PMID- 16467952 TI - Psychotic-like experiences and interpersonal violence in the general population. AB - BACKGROUND: Research on the association of psychopathology and violence has mainly focused on severe but rare mental disorders, especially psychotic disorders. However, evidence is growing that psychotic disorders are continuous with common psychotic-like experiences in the general population. This study aimed to examine the association of psychotic-like experiences with violence in a general population sample. METHODS: In 38,132 adult participants of the 2001 US National Household Survey on Drug Abuse, the association of psychotic-like experiences with violent behavior were examined. RESULTS: Psychotic-like experiences were reported by 5.1% (N = 2,584) of adults in the community. These experiences were associated with increased risk of attacking someone with the intent of hurting that person (Odds Ratio [OR] = 5.72), intimate partner violence (OR = 4.97), arrests for aggravated assault (OR = 5.12), and arrests for other assault (OR = 3.65). The risk of violence increased with the number of psychotic like experiences. Unusual perceptual experiences and paranoid ideations were more consistently associated with violence. CONCLUSIONS: The link between psychopathology and interpersonal violence appears to expand beyond the limits of severe mental disorders and to include more common psychotic-like experiences in the general population. PMID- 16467953 TI - Early predictors of parent- and self-reported perceived global psychological difficulties among adolescents: a prospective cohort study from age 3 to age 15. AB - AIMS: To study predictors at age 3 and at age 12 for parent and self-reported perceived global psychological difficulties at age 15. METHOD: A representative birth cohort was prospectively followed from early childhood to age 15. Ratings of children's behavioral and emotional problems were collected at age 3 (Child Behavior Check List 2/3) and at age 12 (Child Behavior Check List 4-16, Youth Self Report). Mothers and fathers separately completed a questionnaire on their own well-being, health and mental distress when the child was 12. At the same time-point, family functioning was measured with the Family Assessment device. Outcome variables included both parent and self-reports of children's perceived psychological difficulties at age 15. RESULTS: At age 15, of the 707 children with both parent and self-reports on perceived difficulties available, 10% had more perceived difficulties than peers of the same age in parent or self-reports. There was a significant increase in perceived difficulties from age 12 to age 15 in self-reports but not in parent reports. The parent-child agreement on difficulties at both time-points was very low (proportion of agreement 0.12 0.17). At age 3, externalizing problems, especially aggressiveness, predicted parent reports of child's difficulties at 15 years of age in univariate analysis. At age 12, parent-reported child's externalizing symptoms and perceived difficulties, poor social competence in self-reports, and mothers' reports of her own poor well-being, independently predicted parents' reporting of their child's perceived difficulties at age 15. Self-reports of internalizing problems and mother's own reports of her depressiveness when the child was 12 independently predicted the 15-year old's self-reported difficulties. Furthermore, in univariate analysis, poor family functioning at age 12 predicted perceived difficulties in parent and self-reports. CONCLUSIONS: Both parent and self reports of a child's difficulties are the outcome of an accumulation of the child's psychopathology, parental distress, and family dysfunction. The results emphasize the importance of early detection of children at risk of negative developmental pathways. In clinical practice and in prevention interventions, it is important to take into account the child's individual psychopathology, parental distress, and family dysfunction. PMID- 16467954 TI - Mental health profiles among married, never-married, and separated/divorced mothers in a nationally representative sample. AB - BACKGROUND: Several studies have found that married mothers compared to single mothers had better mental health (Cairney et al. in Soc Psychiatry Psychiatr Epidemiol 38:442-449, 2003; Cairney et al. in Can J Public Health 90:320-324, 1999; Davies et al. in J Marriage Fam 59:294-308, 1997; Lipman et al. in Am J Psychiatry 158:73-77, 2001; Wang in Soc Psychiatry Psychiatr Epidemiol 39:26-32, 2004). Although a relationship between family structure (single vs married mothers) and psychiatric disorders is well established, several questions remain. The present study addressed the question "Are there differences in the prevalence of psychiatric disorders between married, never-married, and separated/divorced mothers?" METHODS: The present report examined the lifetime prevalence of anxious misery, fear, and externalizing disorders among mothers in relation to family structure (married, never-married, and separated/divorced) in the US National Comorbidity Survey (N=1,534). RESULTS: Results indicated that never-married mothers appeared to be generally similar to married mothers in their mental health profiles. Separated/divorced mothers compared to married mothers had increased odds of having any anxious-misery disorder, depression, dysthymia, generalized anxiety disorder (GAD), posttraumatic stress disorder, any externalizing disorder, and antisocial personality disorder. Differences were found between never-married and separated/divorced mothers, with separated/divorced mothers having increased odds ratios of having any anxious misery disorder, depression, and GAD. CONCLUSIONS: Results are discussed in light of the unique life contexts of married, never-married, and separated/divorced mothers and as further evidence for the case against combining the separated/divorced and never-married marital status into one "single motherhood" classification in mental health research. PMID- 16467955 TI - Comparison of androgen receptor CAG and GGN repeat length polymorphism in humans and apes. AB - Two polymorphic trinucleotide repeats of human androgen receptor gene (hAR), CAG and GGN which encode glutamine and glycine, have been shown to be associated with human diseases. The number of repeats ranges from 8 to 35 for the CAG and from 10 to 30 for the GGN in human populations. Longer CAG repeats are associated with reduced hAR transcriptional activity, spinal bulbar muscular atrophy and lower cognitive function in older men, whereas shorter CAG repeats are associated with increased risk of prostate cancer and infertility in men. The functional roles of the CAG and GGN repeats have not been clarified. In order to compare the sequence of the CAG and GGN regions in apes, we analyzed 57 chimpanzees, 18 gorillas, 20 orangutans, 16 agile gibbons, and 17 siamangs by PCR and electrophoresis. Two bonobos and one long-tailed macaque were also sequenced and the sequences of all species were aligned, respectively, with one human registered sequence. Seventeen different alleles (4, 7, 8, 9, 12, 14, 15, and 17-26 repeats) and 11 alleles (11 14 and 16-22 repeats) were detected at the CAG and the GGN loci, respectively. Although the repeat tract was conserved among apes, chimpanzees had alleles with a wide range of repeat lengths: (CAG)(14-26) and (GGN)(14-22). Gorillas were less polymorphic with the (CAG)(8) and (GGN)(19) alleles being most common, and orangutans exhibited monomorphic (CAG)(11) and (GGN)(22) alleles. On the other hand, agile gibbons and siamangs had the shortest (CAG)(4) allele, but showed variable length of GGN repeats (11-13 in agile gibbons and 16-21 in siamangs). In chimpanzees, frequent haplotypes consisting of short CAG repeats and long GGN repeats or vice versa was observed as in humans. PMID- 16467956 TI - How costly are ejaculates for Japanese macaques? AB - Much sexual selection theory is based on the idea that ejaculate is cheap. Since further details are unknown our aim was to determine the energy that primate males require for ejaculate production. We addressed this problem by measuring the energy content (in kJ) of ejaculates from Japanese macaques (Macaca fuscata) using standard bomb calorimetry. Then, we estimated the relative amount of energy that individuals require for ejaculate production by relating the net energy content of ejaculates to males' daily basal metabolic rate (BMR). Fresh macaque ejaculate contains 3.0 kJ ml(-1). Assuming a mean volume of 2.7 ml an average macaque ejaculate contains 8.1 kJ. Depending on the individuals' body mass (6-13 kg) and the number and volume of the ejaculates, macaque males are assumed to use between at least 0.8% and at most 6.0% of their BMR for ejaculate production per day during the breeding season. Even when regarding only the minimal energy investment of 0.8% of daily BMR for ejaculate production, clearly ejaculates come with some cost for primate males. PMID- 16467957 TI - Red-tailed monkeys (Cercopithecus ascanius) hunt green pigeons (Treron calva) in the Kalinzu Forest in Uganda. AB - Red-tailed monkeys (Cercopithecus ascanius) were observed hunting green pigeons (Treron calva) in the Kalinzu Forest in Uganda. During 2 h 39 min, I observed two cases of successful hunting and one case of unsuccessful hunting in a Ficus saussureana tree. Red-tailed monkeys stalked the pigeons until they were within 2 3 m, and then jumped and caught them. In both successful cases, blue monkeys (C. mitis) ran to the hunting site from adjacent trees in order to poach the prey, and the red-tailed monkeys fled. One of these red-tailed monkeys dropped the pigeon while fleeing, and the blue monkey climbed down from the tree to search for it. This is the first record of cercopithecoid monkeys hunting birds that are outside of the nest and moving freely, and also the first record of red-tailed monkeys hunting vertebrates. However rare it may be, the repeated hunting attempts using similar techniques and the immediate attempt of blue monkeys to poach the prey suggest that this type of hunting was not a one-time event that happened by chance. Blue monkeys and an adult chimpanzee (Pan troglodytes schweinfurthii) in and around the fig tree did not attempt to hunt. The hunting of volant birds may be enabled by the small body size and the quick movements of red-tailed monkeys. PMID- 16467958 TI - Captive female gorilla agonistic relationships with clumped defendable food resources. AB - Minimal feeding competition among female mountain gorillas (Gorilla gorilla beringei) has resulted in egalitarian social relationships with poorly defined agonistic dominance hierarchies. Thus, gorillas are generally viewed as non competitive egalitarian folivores that have had little need to develop effective competitive strategies to access food resources. However, this generalization is inconsistent with more recent research indicating that most gorillas are frugivorous, feeding on patchily distributed food resources. The current study at Howletts Wild Animal Park, Kent, England, explores the effects of clumped and defendable foods on female gorilla agonistic relationships among three groups of western lowland gorillas (G. g. gorilla), conditions that are predicted to lead to well-differentiated agonistic dominance hierarchies among female primates. The Howletts gorillas foraged all day on low-energy/-nutrient, high-fiber foods widely distributed around their enclosure by the keepers. However, they also had periodic access to high-energy foods (e.g., nuts, raisins, strawberries, etc.) that the keepers would spread in a clumped and defendable patch. Frequencies of agonistic and submissive behaviors between females and proximity data were gathered. High-status females were found to monopolize the food patch and kept the low-status females at bay with cough-grunt threat vocalizations or by chasing them away. Agonistic interactions were initiated mostly by females of high status; these were directed towards females of low status and were generally not reciprocal. In addition, females of low status engaged in submissive behaviors the most often, which they directed primarily at females of high status, especially in response to aggression by the latter. Agonistic interactions between high- and low-status females had decided outcomes more often than not, with low-status females the losers. Competition over highly desirable foods distributed in defendable clumps at Howletts appears to have led to well-defined dominance relationships among these female gorillas. PMID- 16467959 TI - Analyses of the factors influencing bone graft infection after delayed cranioplasty. AB - BACKGROUND: Several factors influencing bone graft infection after delayed cranioplasty are analyzed in order to reduce the occurrence of infection. METHODS: For about 10 years, from March 1995 to February 2005, delayed cranioplasty was performed for 206 cases. The cases comprised 124 males and 82 females. Age distribution of the patients ranged from 6 months to 79 years old. The mean postoperative follow-up period was 1834 days. Autogenous bone, which was preserved in 100% ethanol at -20 degrees C and autoclaved before operation, was used in 54 patients. Polymethylmethacrylate (PMMA) was used in 55 patients. Custom-made PMMA was used in 3 patients. Custom-made titanium mesh was used in 77 patients. Custom-made ceramics (Alumina-ceramics 10 cases and hydroxyapatite 7 cases) was used in 17 patients. FINDINGS: Autoclaved and autogenous bone graft and PMMA have a significantly high rate of graft infection. Titanium mesh has the significantly lowest rate of graft infection. Alumina-ceramic has a merit that it has sufficient strength, however the number of cases using custom-made ceramics including alumina-ceramic was relatively small, and thus we cannot find significant differences in infection rate compared with that of other materials. There was no statistically significant difference in the bone graft infection rate among four categories of preceding diseases; cerebrovascular diseases, head trauma, infectious diseases, and brain tumour. CONCLUSION: Autoclaved and autogenous bone grafts and PMMA have a significantly higher rate of graft infection. Titanium mesh has the significantly lowest rate of graft infection. PMID- 16467960 TI - Thoracic myelocystoceles--two variants. AB - Myelocystoceles are rare lesions. Rarer still are thoracic myelocystoceles. Two patients with thoracic myelocystoceles are being reported. The first patient presented with swelling in the thoracic region with paraplegia and incontinence. MR with MR myelography revealed the malformation to be a thoracic myelocystocele. The second patient presented with a midline cutaneous mass lesion without neurological deficit. MRI revealed a Type I split cord malformation, a CSF containing sac in the thoracic region that communicated through a stalk-like structure to the hydromyelic hemicord. Surgery in both these patients did not produce any change in the neurological status. The importance of recognizing thoracic myelocystoceles, their clinical presentation, embryology, value of MR myelography in the diagnosis and role of surgery are discussed. The differences between terminal myelocystoceles and thoracic myelocystoceles are also highlighted. PMID- 16467961 TI - Cerebral blood oxygenation changes induced by bypass blood flow in moyamoya disease and non-moyamoya cerebral ischaemic disease. AB - BACKGROUND: Superficial temporal artery-middle cerebral artery (STA-MCA) anastomosis has been used to prevent stroke in patients with moyamoya disease (MD) and non-moyamoya ischaemic disease (non-MD). However, little is yet known regarding the difference between these groups of patients in the extent to which the bypass contributes to maintaining adequate cerebral blood oxygenation (CBO), or the temporal changes after surgery. In the present study, we evaluated the CBO changes induced by bypass blood flow in patients with MD and non-MD during the peri-operative periods employing optical spectroscopy. METHODS: We investigated 13 patients who underwent STA-MCA anastomosis, including 5 MD and 8 non-MD patients. We evaluated the effects of STA blood flow on the CBO in the MCA territory on the anastomosis side, employing visual light spectroscopy during surgery and near infrared spectroscopy (NIRS) at one week after surgery. FINDINGS: In 4 MD patients and one non-MD patient, the STA blood flow increased the oxyhaemoglobin and cortical oxygen saturation (CoSO2), indicating that the bypass supplied blood flow to the ischaemic brain; the CBO changes were observed more frequently in MD than in non-MD patients (p<0.02). The pre-anastomosis CoSO2 (65.4+/-5.4%) in MD was significantly lower than that (72.8+/-7.6%) in non-MD (p<0.05). Postoperative NIRS demonstrated that the bypass began to supply blood flow to the brain in 5 non-MD patients whose bypass did not supply blood flow during surgery. CONCLUSIONS: Although MD has vessels of small diameter as compared to non-MD, the bypass begins to supply blood flow to the ischaemic brain earlier in MD than in non-MD after anastomosis. The fact that the CoSO2 in MD was lower than that in non-MD suggested that the perfusion pressure in MD was lower than that in non-MD, and this might account for the difference in the bypass blood supply after anastomosis between MD and non-MD. Our data suggest that, even if the bypass does not supply blood to the brain during surgery in non-MD, the bypass blood flow gradually increases after surgery. PMID- 16467962 TI - The enigmatic origin of subfrontal schwannomas: report of a case without hyposmia. AB - Subfrontal schwannomas are rare intracranial tumors. Most of them are associated with hyposmia/anosmia. The source of origin of these tumors is still incompletely understood. We report a 23-year-old male who presented with recurrent focal motor seizures, but had no hyposmia. The tumor was completely removed by a subfrontal approach. Relevant literature has been reviewed. PMID- 16467963 TI - Meta analyses in treatment of spontaneous supratentorial intracerebral haematoma. AB - BACKGROUND: None of the randomized controlled trials (RCT) on treatment of Intracerebral haematoma (ICH), definitely shows surgery to be beneficial over conservative treatment alone. Systematic reviews that pooled these RCTs were also inconclusive. This systematic review updates previous meta-analyses, using an alternative manner of reviewing with a criteria list constructed specifically for this type of disease and related interventions. METHODS: RCTs and quasi-RCTs (q RCT) published in English were identified with a systematic literature search. They were evaluated with disease/intervention-specific criteria on comparability between intervention and control group concerning prognostic factors, co interventions and effect measurement. The resulting selection of studies was compared with those of two earlier systematic reviews. In a meta-analysis selected studies were statistically pooled. FINDINGS: The meta-analysis of surgery versus conservative treatment failed to show a statistically significant reduction in the odds of death (OR: 0.84, 95% CI: 0.67-1.07) in surgically treated patients. CONCLUSIONS: Like previous reviews, our disease/intervention specific methodological evaluation showed no reduction in mortality. Sensitivity analysis demonstrates that the manner in which studies are methodologically evaluated in a systematic review has a great impact on its conclusions. PMID- 16467965 TI - Development and mathematical modeling of a two-stage reactor system for trichloroethylene degradation using Methylosinus trichosporium OB3b. AB - A two-stage reactor system was developed for the continuous degradation of gas phase trichloroethylene (TCE). Methylosinus trichosporium OB3b was immobilized on activated carbon in a TCE degradation reactor, trickling biofilter (TBF). The TBF was coupled with a continuous stirred tank reactor (CSTR) to allow recirculation of microbial cells from/to the TBF for the reactivation of inactivated cells during TCE degradation. The mass transfer aspect of the TBF was analyzed, and mass transfer coefficient of 3.9 h(-1) was estimated. The loss of soluble methane monooxygenase (sMMO) activity was modeled based on a material balance on the CSTR and TBF, and transformation capacity (T (c)) was determined to be 20.2 micromol mg(-1). Maximum TCE degradation rate of 525 mg 1(-1) d(-1) was obtained and reactor has been stably operated for more than 270 days. PMID- 16467966 TI - Kinetic analysis of the inhibitory effect of trichloroethylene (TCE) on nitrification in cometabolic degradation. AB - In this study, the inhibitory effect of TCE on nitrification process was investigated with an enriched nitrifier culture. TCE was found to be a competitive inhibitor of ammonia oxidation and the inhibition constant (K(I)) was determined as 666-802 microg/l. The TCE affinity for the AMO enzyme was significantly higher than ammonium. The effect of TCE on ammonium utilization was evaluated with linearized plots of Monod equation (e.g., Lineweaver-Burk, Hanes Woolf and Eadie-Hofstee plots) and non-linear least square regression (NLSR). No significant differences were found among these data evaluation methods in terms of kinetic parameters obtained. PMID- 16467964 TI - Regulation and expression of heme oxygenase enzymes in aged-rat brain: age related depression in HO-1 and HO-2 expression and altered stress-response. AB - The heme oxygenase isozymes, HO-1 and HO-2, oxidatively cleave the heme molecule to produce biliverdin and the gaseous messenger, CO. The cleavage results in the release of iron, a regulator of transferrin, ferritin, and nitric oxide (NO) synthase gene expression. Biliverdin reductase (BVR) then catalyzes the reduction of biliverdin, generating the potent intracellular antioxidant, bilirubin. We report an age-related decrease in HO-1 and HO-2 expression present in select brain regions including the hippocampus and the substantia nigra, that are involved in the high order cognitive processes of learning and memory. The age related loss of monoxide-producing potential in select regions of the brain was not specific to the HO system but was also observed in neuronal NO-generating system. Furthermore, compared to 2-month old rats, the ability of aged brain tissue to respond to hypoxic/hyperthermia was compromised at both the protein and the transcription levels as judged by attenuated induction of HO-1 immunoreactive protein and its 1.8 Kb transcript. Neotrofin (AIT), a cognitive-enhancing and neuroprotective drug, caused a robust increase in HO-1 immunoreactive protein in select neuronal regions and increased the expression of HO-2 transcripts. The potential interplay between regulation of HO-2 gene expression and the serum levels of the adrenal steroids is discussed. We suggest the search for therapeutic agents that reverse the decline and aberrant stress response of HO enzymes may lead to effective treatment regimens for age-associated neuronal deficits. PMID- 16467967 TI - Biodegradation kinetics of linear alkylbenzene sulphonates in sea water. AB - This article reports the primary biodegradation kinetics of linear alkylbenzene sulphonates (LAS) in sea water from the Bay of Cadiz (South West of the Iberian Peninsula). The authors used the biodegradation test guideline proposed by the Office of Prevention, Pesticides, and Toxic Substances of the United States Environmental Protection Agency; 835.3160 "Biodegradability in sea water" in its shake flask variant. High performance liquid chromatography (HPLC) has been employed for the analysis of the surfactant material. The surfactant shows a primary biodegradation kinetic in accordance with a logistic model, the kinetic parameters t (50) and lag time were calculated by means of a easy quantitative procedure introduced. Mean values of 6.15 +/- 0.45 and 6.67 +/- 0.6 days were obtained for t (50) and lag time, respectively. These results indicate that although LAS has a high primary biodegradation rate in sea water, it biodegrades slower than in similar tests conducted in river water. PMID- 16467970 TI - Fluorescence decays and photon propagation times. AB - The effect of the time of flight of the exciting and emitted photons on fluorescence decays is studied. Experimental results for a long lifetime molecule (coronene) are analysed according to the fluorescence decay law previously obtained by the authors. The developed model accounts well for the main features of the observations. The inclusion of photon propagation times is essential for a correct description of the fluorescence decays under the described circumstances. PMID- 16467968 TI - ERK regulation upon contact inhibition in fibroblasts. AB - Despite the understanding of the importance of mitogen-activated protein (MAP) kinase activation in the stimulation of growth, little is known about the role of MAP kinase regulation during contact inhibited growth control. To investigate the role of the MAP kinase extracellular signal-regulated kinase (ERK) during the transition to a contact inhibited state, cultures of normal fibroblasts (BJ) were grown to different stages of confluency. The levels of MAP kinase phosphatase (MKP) expression and the amount of active ERK and MAP ERK kinase (MEK) in these cultures were assessed through western blot analysis and were compared to fibrosarcoma cell cultures (HT-1080), which lack contact inhibition. In normal fibroblasts, the amounts of active MEK and ERK decline at contact inhibition, concurrently with a rise in MKP-1, MKP-2, and MKP-3 protein levels. In contrast, fibrosarcoma cells appear to lack density-dependent regulation of the ERK pathway. Additionally, altering the redox environment of fibrosarcoma cells to a less reducing state, as seen during contact inhibition, results in increased MKP 1 expression. Taken together, these results suggest that the altered redox environment upon contact inhibition may contribute to the regulation of ERK inactivation by MKPs. PMID- 16467969 TI - The influence of human serum albumin on the photogeneration of singlet oxygen by meso-tetra(4-sulfonatophenyl)porphyrin. An infrared phosphorescence study. AB - meso-Tetra(4-sulfonatophenyl)porphyrin (TPPS4) is a water soluble photosensitizer, which is currently clinically tested as a PDT drug. In our contribution, we present IR spectral- and time-resolved phosphorescence data reflecting the influence of human serum albumin (HSA) on singlet oxygen photogeneration by TPPS4. IR emission of TPPS4 was studied in samples containing various concentrations of HSA in phosphate buffer. The observed changes in spectral and temporal behaviour of TPPS4 and singlet oxygen phosphorescence caused by the addition of HSA are equivalent to the effect of nitrogen purging of HSA-free solutions of TPPS4. The main feature induced by addition of HSA appears to be the occurrence of a long-lived (tens of microseconds) photosensitizer phosphorescence at 900 nm besides ordinary short-lived (approximately 2 micros) one at 820 nm. It is accompanied by presence of a long-lived component of singlet oxygen emission with lifetime roughly corresponding to that of the long photosensitizer phosphorescence component. Moreover, the quantum yield of singlet oxygen phosphorescence decreases with increasing HSA concentration, while total quantum yield of TPPS4 phosphorescence rises. These facts are explained by a shielding effect of HSA on bound molecules of TPPS4 against quenching by oxygen which is analogous to oxygen removal by nitrogen purging. PMID- 16467971 TI - Laparoscopic Fowler-Stephens procedure is contraindicated for intraabdominal testicular major duct anomalies. AB - The laparoscopic one- and two-stage Fowler-Stephens procedure has gained large popularity in the treatment of the child with a high intraabdominal testis. It is largely debated which technique (e.g., testicular autotransplant or Fowler Stephens procedure) offers the best results for high intraabdominal testes. We describe a case of a 3-year-old boy for whom previous bilateral inguinal exploration results were negative for testes or testicular remnants. The diagnostic laparoscopy showed two iliac intraabdominal testes with short spermatic vessels, closed inguinal rings, and complete dissociation of didime epididime. A left open orchidopexy was perfomed, and testicular autotransplant was proposed for the right testes located 4 cm from the internal inguinal ring. Long-term follow-up evaluation (1.8 years) of the left testis showed it in the scrotum with good testicular size (1.5 cm). We believe that there are two main reasons to contraindicate the Fowler-Stephens technique: associated malformation that does not permit the development of the collateral blood flow via the vasal artery necessary for a viable testis, and previous surgery that represents a risk factor for testicular atrophy. The laparoscopic anatomic classification for the intraabdominal testis is reliable and can disclose the most suitable surgical technique. Laparoscopy is a valuable tool in the diagnosis and treatment of the nonpalpable testicle. PMID- 16467972 TI - Daily physical education in the school curriculum in prepubertal girls during 1 year is followed by an increase in bone mineral accrual and bone width--data from the prospective controlled Malmo pediatric osteoporosis prevention study. AB - The aim of this study was to evaluate a general school-based 1-year exercise intervention program in a population-based cohort of girls at Tanner stage I. Fifty-three girls aged 7-9 years were included. The school curriculum-based exercise intervention program included 40 minutes/school day. Fifty healthy age matched girls assigned to the general school curriculum of 60 minutes physical activity/week served as controls. Bone mineral content (BMC, g) and areal bone mineral density (aBMD, g/cm(2)) were measured with dual X-ray absorptiometry (DXA) of the total body (TB), lumbar spine (L2-L4 vertebrae), third lumbar vertebra (L3), femoral neck (FN), and leg. Volumetric bone mineral density (g/cm(3)) and bone width were calculated at L3 and FN. Total lean body mass and total fat mass were estimated from the TB scan. No differences at baseline were found in age, anthropometrics, or bone parameters when the groups were compared. The annual gain in BMC was 4.7 percentage points higher in the lumbar spine and 9.5 percentage points higher in L3 in cases than in controls (both P < 0.001). The annual gain in aBMD was 2.8 percentage points higher in the lumbar spine and 3.1 percentage points higher in L3 in cases than in controls (both P < 0.001). The annual gain in bone width was 2.9 percentage points higher in L3 in cases than in controls (P < 0.001). A general school-based exercise program in girls aged 7-9 years enhances the accrual of BMC and aBMD and increases bone width. PMID- 16467973 TI - Structural analysis of trabecular bone of the proximal femur using multislice computed tomography: a comparison with dual X-ray absorptiometry for predicting biomechanical strength in vitro. AB - We investigated whether trabecular microstructural parameters determined in multislice spiral computed tomographic (MSCT) images of proximal femur specimens differed in male and female donors and improved the prediction of biomechanical strength of the femur compared to bone mineral density (BMD) and content (BMC) determined with dual X-ray absorptiometry (DXA) as the standard diagnostic technique. Proximal femur specimens (n = 119) were harvested from formalin-fixed human cadavers (mean age 80 +/- 10 years). BMD was determined using DXA. Trabecular microstructural parameters (bone volume fraction, fractal dimension, and trabecular thickness, spacing, and number) were calculated in MSCT-derived images of the proximal femur. Failure load (FL) was measured using a biomechanical side-impact test. An age-, height-, and weight-matched subgroup (n = 54) was chosen to compare male and female donors. BMC, BMD, and structural parameters correlated significantly with FL, with r up to 0.75, 0.71, and 0.71, respectively. In a multiple regression model, an increase up to r = 0.82 was obtained when combining trabecular structural parameters and BMC. BMD differed between males and females only at the trochanter. BMC showed significant gender differences in all regions. This experimental study showed that a combination of BMC and microstructural parameters could improve the prediction of FL, suggesting that bone mass and trabecular structure carry overlapping but complementary information and that a combination of the two provides the best prediction of bone strength. Male donors had larger femora even after adjustment for body size and height, but no differences in trabecular structure were found between males and females. PMID- 16467975 TI - Mortality rates of patients with a hip fracture in a southwestern district of Greece: ten-year follow-up with reference to the type of fracture. AB - Increased mortality after a hip fracture has been associated with age, sex, and comorbidity. In order to estimate the long-term mortality with reference to hip fracture type, we followed 499 patients older than 60 years who had been treated surgically for a unilateral hip fracture for 10 years. At admission, patients with femoral neck fractures (n = 172) were 2 years younger than intertrochanteric patients (77.6 +/- 7.7 [SD] vs. 79.9 +/- 7.4 [SD], P = 0.001) and had a greater prevalence of heart failure (57% vs. 40.3%, P = 0.03). Similar mortality rates were observed at 1 year in both types of fracture (17.9% vs. 11.3%, log rank test P = 0.112). Mortality rates were significantly higher for intertrochanteric fractures at 5 years (48.8% vs. 34.7%, P = 0.01) and 10 years (76% vs. 58%, P = 0.001). Patients 60-69 years old with intertrochanteric fractures had significantly higher 10-year mortality than patients of similar age with femoral neck fractures (P = 0.008), while there was no difference between the groups aged 70-79 (P > 0.3) and 80-89 (P = 0.07). Women were less likely to die in 5 years (relative risk [RR] = 0.57, 95% confidence interval [CI] 0.41-0.79, P = 0.0007) and 10 years (RR = 0.65, 95% CI 0.49-0.85, P = 0.002). Age, sex, the type of fracture, and the presence of heart failure were independent predictors of 10 year mortality (Cox regression model P < 0.0001). The intertrochanteric type was independently associated with 1.37 (95% CI 1.03-1.83) times higher probability of death at 10 years (P = 0.002). In conclusion, the type of fracture is an independent predictor of long-term mortality in patients with hip fractures, and the intertrochanteric type yields worse prognosis. PMID- 16467974 TI - Regulation of cementoblast gene expression by inorganic phosphate in vitro. AB - Examination of mutant and knockout phenotypes with altered phosphate/pyrophosphate distribution has demonstrated that cementum, the mineralized tissue that sheathes the tooth root, is very sensitive to local levels of phosphate and pyrophosphate. The aim of this study was to examine the potential regulation of cementoblast cell behavior by inorganic phosphate (P(i)). Immortalized murine cementoblasts were treated with P(i) in vitro, and effects on gene expression (by quantitative real-time reverse-transcriptase polymerase chain reaction [RT-PCR]) and cell proliferation (by hemacytometer count) were observed. Dose-response (0.1-10 mM) and time-course (1-48 hours) assays were performed, as well as studies including the Na-P(i) uptake inhibitor phosphonoformic acid. Real time RT-PCR indicated regulation by phosphate of several genes associated with differentiation/mineralization. A dose of 5 mM P(i) upregulated genes including the SIBLING family genes osteopontin (Opn, >300% of control) and dentin matrix protein-1 (Dmp-1, >3,000% of control). Another SIBLING family member, bone sialoprotein (Bsp), was downregulated, as were osteocalcin (Ocn) and type I collagen (Col1). Time-course experiments indicated that these genes responded within 6-24 hours. Time-course experiments also indicated rapid regulation (by 6 hours) of genes concerned with phosphate/pyrophosphate homeostasis, including the mouse progressive ankylosis gene (Ank), plasma cell membrane glycoprotein-1 (Pc 1), tissue nonspecific alkaline phosphatase (Tnap), and the Pit1 Na-P(i) cotransporter. Phosphate effects on cementoblasts were further shown to be uptake dependent and proliferation-independent. These data suggest regulation by phosphate of multiple genes in cementoblasts in vitro. During formation, phosphate and pyrophosphate may be important regulators of cementoblast functions including maturation and regulation of matrix mineralization. PMID- 16467976 TI - Osteoblast dysfunction in male idiopathic osteoporosis. AB - The etiology of primary osteoporosis in young and middle-aged men is unknown. We have studied osteoblast function in cells derived from men with idiopathic osteoporosis and in control cells from age-matched men with osteoarthrosis. Osteoblasts were isolated from transiliac bone biopsies. Osteoblast function was measured as vitamin D-stimulated osteocalcin production and production of cytokines and factors involved in osteoclast activation and bone formation. Cell proliferation was measured as (3)H-thymidine incorporation. Parathyroid hormone related peptide (PTHrP) mRNA was measured using reverse-transcriptase polymerase chain reaction. In osteoporotic men, bone mineral density at the femoral neck was correlated to in vitro production of osteocalcin. Osteoblasts from osteoporotic men produced significantly less osteocalcin after vitamin D stimulation but had increased production of macrophage colony-stimulating factor (M-CSF) compared to controls. The osteocalcin response was negatively correlated to production of M CSF, interleukin-6, and C-terminal propeptide of type I collagen. Basal (3)H thymidine incorporation was similar in cells from osteoporotic patients and controls. PTHrP (10(-9 )M) significantly increased cell proliferation in control cells but not in osteoporotic cells. Basal PTHrP mRNA levels were significantly higher in osteoporotic cells than in cells from controls. The results are in agreement with previous histomorphologic studies indicating that men with idiopathic osteoporosis have an osteoblast dysfunction with decreased osteocalcin production and increased production of factors stimulating osteoclast activation. This indicates a catabolic cellular metabolic balance leading to negative bone turnover, resulting in osteoporosis. The cause of such cellular dysfunction needs further evaluation. PMID- 16467977 TI - B cells play an important role in lipopolysaccharide-induced bone resorption. AB - The host immune system, especially activated T cells, plays a crucial role in inflammatory bone resorption and osteoclastogenesis. Previously, we showed that T cells are involved in inflammatory bone resorption in vivo. However, little is known about whether B cells are involved in inflammatory bone resorption and how B cells take part in osteoclastogenesis. Therefore, the aim of this study was to examine whether B c ells truly influence inflammatory bone resorption in vivo. Alveolar bone resorption in normal mice, in SCID mice that lack both B and T cells, and in B cell-reconstituted SCID mice was compared histopathologically after repeated injections of lipopolysaccharide (LPS) into the gingiva. Furthermore, we examined whether the B cells that are stimulated by LPS are involved in osteoclastogenesis in vitro. As a result, the B cell-reconstituted SCID mice showed stronger inflammatory bone resorption than the SCID mice. Also, in vitro, LPS-stimulated B cells enhanced osteoclastogenesis and anti-tumor necrosis factor (TNF)-alpha antibody completely blocked osteoclastogenesis induced by LPS-stimulated B cells. These results suggest that B cells promote inflammatory bone resorption through TNF-alpha. PMID- 16467978 TI - Gene expression of runx2, Osterix, c-fos, DLX-3, DLX-5, and MSX-2 in dental follicle cells during osteogenic differentiation in vitro. AB - Recently, osteogenic precursor cells were isolated from human dental follicles, which differentiate into cementoblast- or osteoblast- like cells under in vitro conditions. However, mechanisms for osteogenic differentiation are not known in detail. Dental follicle cell long-term cultures supplemented with dexamethasone or with insulin resulted in mineralized nodules, whereas no mineralization or alkaline phosphatase activity was detected in the control culture without an osteogenic stimulus. A real-time reverse-transcriptase polymerase chain reaction (PCR) analysis was developed to investigate gene expression during osteogenic differentiation in vitro. Expression of the alkaline phosphatase (ALP) gene was detected during differentiation in the control culture and was similar to that in cultures with dexamethasone and insulin. DLX-3, DLX-5, runx2, and MSX-2 are differentially expressed during osteogenic differentiation in bone marrow mesenchymal stem cells. In dental follicle cells, gene expression of runx2, DLX 5, and MSX-2 was unaffected during osteogenic differentiation in vitro. Osteogenic differentiation appeared to be independent of MSX-2 expression; the same was true of runx2 and DLX-5, which were protagonists of osteogenic differentiation and osteocalcin promoter activity in bone marrow mesenchymal stem cells. Like in bone marrow-derived stem cells, DLX-3 gene expression was increased in dental follicle cells during osteogenic differentiation but similar to control cultures. However, gene expression of osterix was not detected in dental follicle cells during osteogenic differentiation; this gene is expressed during osteogenic differentiation in bone marrow stem cells. These real-time PCR results display molecular mechanisms in dental follicle precursor cells during osteogenic differentiation that are different from those in bone marrow-derived mesenchymal stem cells. PMID- 16467979 TI - Characteristics of perianal abscess and fistula-in-ano in healthy children. AB - BACKGROUND: Probably because of the low frequency, perianal abscess (PA) and fistula-in-ano (FIA) in children older than 2 years have not been investigated except in those with a predisposing condition such as Crohn's disease. This study aims to summarize our experience about the characteristics and treatment of PA and FIA in healthy children. METHODS: The charts of all children older than 24 months of age treated for PA and/or FIA from 1990 to 2003 were reviewed. RESULTS: We found 40 patients, 37 of them boys (92.5%), ranging from 2 to 14 years of age (average: 7.19 years). At the first examination, the diagnosis was PA in 36 patients (mean age: 6.8 years; range: 2.3-13 years), and FIA in 4 patients (mean age: 10.8 years; range: 6-14 years). The primary local treatment of PA was drainage (needle aspiration in 26 patients, and incision and drainage in 4 patients) and local care in 6 patients. All patients received antibiotics. Overall, 29 children (80.6%) had primary cure of the abscess. Evolution included recurrent abscess in 3 patients (8.3%) and FIA in 4 patients (11.1%). Crohn's disease was diagnosed in only one boy with an abscess of long duration. No patient developed a new PA in another location or a recurrent FIA. Four male patients aged 6 to 14 years (range: 7.1 years) had a FIA of long duration. One patient underwent a fistulectomy. Crohn's disease was found in three other children and treated conservatively. CONCLUSION: Drainage of PA by needle aspiration associated with antibiotics is effective in children older than 2 years of age with a low rate of evolution toward FIA. Associated pathology must be ruled out in children with FIA. PMID- 16467980 TI - Bilateral adrenalectomy for ectopic Cushing's syndrome-discussions on technique and indication. AB - BACKGROUND: Tumors producing adrenocorticotropic hormone (ACTH) or corticotropin releasing hormone (CRH) often remain undiagnosed until severe Cushing's syndrome appears, and it may be difficult to distinguish from Cushing's syndrome due to pituitary tumors. Many patients suffer from disease spread, with metastases in the liver or other locations, and the main symptoms may be mineral disturbances, diabetes mellitus, or psychological symptoms from the severe hypercortisolism. Bilateral adrenalectomy may alleviate this situation, but is sometimes a troublesome procedure in these severely ill patients. METHODS: We have retrospectively investigated 8 patients with ectopic Cushing's syndrome who have undergone bilateral adrenalectomy at the University Hospital in Uppsala. In addition, another 5 patients who underwent bilateral adrenalectomy for other reasons (recurrent pituitary Mb Cushing or bilateral hyperplasia) were scrutinized for technical considerations. Indications, timing of surgery, and operative procedures were studied to identify signs that may support our approach to management in the future. RESULTS: Curative surgery was not possible in any of the cases with ectopic Cushing's syndrome. Of the 13 operated patients, handport assisted laparoscopic adrenalectomy was successfully performed bilaterally in 5 patients and unilaterally in combination with contralateral open surgery in 1 patient; conventional open surgery was performed on 7 patients, 3 of which were conversions from intitial handport-assisted procedures. Non-fatal complications occurred in 4 out of 10 patients. CONCLUSIONS: We conclude that bilateral handport-assisted laparoscopic adrenalectomy is safe, and that all surgical techniques in these severely ill patients may be troublesome and technically demanding. Early surgical intervention may reduce the technical disadvantages. Moreover, bilateral adrenalectomy can substantially reduce the symptoms of Cushing's syndrome, although effects on mortality are not obvious. PMID- 16467981 TI - Correlations between vitamin D status and biochemical/clinical and pathological parameters in primary hyperparathyroidism. AB - BACKGROUND: To determine the prevalence of vitamin D deficiency and the effects of vitamin D status on parathyroid adenoma weight, clinical and biochemical indices in patients with primary hyperparathyroidism (pHPT) were studied. METHODS: Eighty patients with pHPT who underwent surgical treatment and in whom the presence of parathyroid adenoma were confirmed histopathologically were studied retrospectively from recorded data files. Patients were divided into three groups: patients with 25-hydroxyvitamin D (25-OHD) concentrations < 15 ng/ml (group 1, n = 44), patients with 25-OHD concentrations > 15-25 ng/ml (group 2, n = 9), and patients with 25-OHD concentrations > 26 ng/ml (group 3, n = 27). Serum calcium, phosphate, alkaline phosphatase, creatinine, and albumin levels and urinary calcium excretion were determined by auto-analyzer. Plasma 25-OHD and parathyroid hormone (PTH) levels were determined by immunoradiometric assay using commercially available kits. RESULTS: No statistically significant differences were observed with respect to serum calcium, phosphorus, albumin, and creatinine concentrations between these groups. Serum PTH, alkaline phosphatase concentrations, urinary calcium excretion, parathyroid adenoma weight, and postoperative sixth month PTH concentrations were significantly higher in group 1 patients than in group 2 and group 3 patients. Significant correlations were observed between parathyroid adenoma weight and serum 25-OHD concentrations (r = 0.348, P = 0.020); parathyroid adenoma weight and urinary calcium excretion (r = 0.348, P = 0.021). Multiple regression analysis revealed that parathyroid adenoma weight, serum 25-OHD, and preoperative PTH concentrations correlated independently and significantly with postoperative sixth month PTH concentrations. CONCLUSIONS: Vitamin D deficiency leads to more severe bone disease, increased parathyroid tumor growth, and delayed postoperative recovery of parathyroid function in patients with primary hyperparathyroidism. PMID- 16467982 TI - Stapler hepatectomy is a safe dissection technique: analysis of 300 patients. AB - BACKGROUND: In many surgical procedures, stapling devices have been introduced for safety and to reduce the overall operative time. Their use for transection of hepatic parenchyma is not well established. Thus, the feasibility of stapler hepatectomy and a risk analysis of surgical morbidity based on intraoperative data have been prospectively assessed on a routine clinical basis. MATERIALS AND METHODS: From October 1, 2001, to January 31, 2005, a total of 416 patients underwent liver resection in our department. During this period endo GIA vascular staplers were used for parenchymal transection in 300 cases of primary (22%) and metastatic (57%) liver cancer, benign diseases (adenoma, focal nodular hyperplasia [FNH], cysts) (14%), gallbladder carcinoma (2%), and other tumors (5%). There were 193 (64%) major resections (i.e., removal of three segments or more) and 107 minor hepatic resections. Additional extrahepatic resections were performed in 44 (15%) patients. RESULTS: Median values for operative time and intraoperative hemorrhage were 210 minutes and 700 ml, respectively. Further, transfusion of RBC and FFP was needed in 17% and 11% of patients, respectively. A postoperative ICU stay for >2 days was required in 18% of patients. The median postoperative hospital stay was 10 days (IQR 8-14 days). The most frequent surgical complications were bile leak (8%), wound infection (3%), and pneumothorax (2%). In 7% of cases after stapler hepatectomy a relaparotomy was necessary. Treated medical complications were pleural effusion (7%), renal insufficiency (5%), and cardiac insufficiency (3%). Risk assessment revealed that both operative time and indication for resection had significant impact on surgical morbidity. Mortality (4%) and morbidity (33%) were comparable to other high-volume centers performing conventional liver resection techniques. CONCLUSION: In conclusion, stapler hepatectomy can be used in a routine clinical setting with a low incidence of surgical complications. PMID- 16467983 TI - Optimal abdominal incision for partial hepatectomy: increased late complications with Mercedes-type incisions compared to extended right subcostal incisions. AB - INTRODUCTION: The optimal abdominal incision for partial hepatectomy has not been established. METHODS: A prospective hepatobiliary surgery database was retrospective reviewed. Patients with Mercedes and extended right subcostal (ERSC) incisions were identified and compared. RESULTS: Between December 1991 and September 2001 a total of 1426 patients met the inclusion criteria. Among them, 856 (60%) had a Mercedes incision and 570 (40%) an ERSC incision. The two groups were well matched for demographics and operative variables. Perioperative morbidity and pulmonary complications were similar for the two groups as well. There was no difference in terms of early wound complications, although incisional hernias occurred in 9.8% of patients with a Mercedes incision compared to 4.8% of those with an ERSC incision (P = 0.0001). On multivariate analysis, the incision type, along with gender, body mass index, and age, were significant predictors of incisional hernia. CONCLUSIONS: An ERSC incision for partial hepatectomy provides adequate, safe access and is associated with fewer long-term wound complications. PMID- 16467985 TI - Duplication, balancing selection and trans-species evolution explain the high levels of polymorphism of the DQA MHC class II gene in voles (Arvicolinae). AB - Major histocompatibility complex (MHC) genes play important role in host-parasite interactions and parasites are crucial factors influencing the population dynamics of hosts. We described the structure and diversity of exon 2 of the MHC class II DQA gene in three species of voles (Arvicolinae) exhibiting regular multi-annual fluctuations of population density and analysed the processes leading to the observed MHC polymorphism. By using cloning-sequencing methodology and capillary electrophoresis-single strand conformation polymorphism, we described seven sequences in the water, eight in the common, and seven in the bank voles coming from an area of 70 km(2) around the Nozeroy canton in the Jura Mountains (Franche Comte, France). All exon 2 sequences translate to give unique amino acid sequences and positive selection was found to act very intensively on antigen binding sites. We documented the presence of recombination at vole DQA region but its importance in generating allelic polymorphism seems to be relatively limited. For the first time within rodents, we documented the duplication of the DQA gene in all three species with both copies being transcriptionally active. Phylogenetic analysis of allelic sequences revealed extensive trans-species polymorphism within the subfamily although no alleles were shared between species in our data set. We discuss possible role of parasites in forming the recent polymorphism pattern of the DQA locus in voles. PMID- 16467986 TI - Identification and expression analysis of interferon gamma genes in channel catfish. AB - Multiple species of type I interferon (IFN) were recently identified in catfish (CF) (Ictalurus punctatus). Herein we extend these studies and report the existence of two distinct type II IFN genes in channel CF. As with zebrafish and the green spotted pufferfish, the two CF IFN-gamma genes are dissimilar in sequence but closely linked on the same chromosome. One of the genes (IFN-gamma2) encodes two distinct messages that likely arose via alternative splicing at two closely spaced splice donor sites within the first intron. Sequence analysis indicates that CF IFN-gamma genes contain the hallmarks of authentic IFN-gamma including: (1) a conserved nuclear localization site at the C terminus (CF IFN gamma2 only), (2) an IFN-gamma signature sequence, (3) six putative helical regions within the mature protein, (4) one or more potential glycosylation sites, and (5) multiple mRNA instability motifs within the 3' untranslated region. Moreover, well-characterized CF T and NK cell clones were shown to synthesize IFN gamma transcripts. This is the first unequivocal demonstration in any lower vertebrate species that NK and T cells synthesize IFN-gamma and is consistent with results in mammalian systems where T cells and NK cells are the major sources of type II IFN production. Collectively, these studies indicate that Siluriformes possess two evolutionarily conserved IFN-gamma genes and demonstrate that CF possess three key elements of the innate immune response: NK cells and types I and II IFN. PMID- 16467987 TI - Immune-related, lectin-like receptors are differentially expressed in the myeloid and lymphoid lineages of zebrafish. AB - The identification of C-type lectin (Group V) natural killer (NK) cell receptors in bony fish has remained elusive. Analyses of the Fugu rubripes genome database failed to identify Group V C-type lectin domains (Zelensky and Gready, BMC Genomics 5:51, 2004) suggesting that bony fish, in general, may lack such receptors. Numerous Group II C-type lectin receptors, which are structurally similar to Group V (NK) receptors, have been characterized in bony fish. By searching the zebrafish genome database we have identified a multi-gene family of Group II immune-related, lectin-like receptors (illrs) whose members possess inhibiting and/or activating signaling motifs typical of Group V NK receptors. Illr genes are differentially expressed in the myeloid and lymphoid lineages, suggesting that they may play important roles in the immune functions of multiple hematopoietic cell lineages. PMID- 16467989 TI - Biotransformation of citrinin to decarboxycitrinin using an organic solvent tolerant marine bacterium, Moraxella sp. MB1. AB - Organic solvent tolerant microorganisms (OSTMs) are novel group of extremophilic microorganisms that have developed resistance to withstand solvent toxicity. These organisms play an important role in biotransformation of organic compounds. In the present study, we used an organic solvent-tolerant marine bacterium, Moraxella sp. MB1. 16S rRNA sequencing revealed that the bacterium shows 98% similarity with an uncultured marine bacterium with GenBank accession no. AY936933. This bacterium was used for the transformation of a toxin, citrinin, into decarboxycitrinin in a biphasic system. This transformation was affected by decarboxylase enzyme produced by MB1. Transformation of citrinin to decarboxycitrinin was monitored by thin-layer chromatography (TLC) and spectrophotometrically. Citrinin decarboxylase activity responsible for transformation was studied in cell-free growth medium and cell lysate of Moraxella sp. MB1. Citrinin decarboxylase was found to be intracellular in nature. The biotransformed product was purified and identified as decarboxycitrinin using electrospray ionization mass spectrometry (ESI-MS/MS) and nuclear magnetic resonance (NMR) spectrometry. The antibiotic activity of both citrinin and decarboxycitrinin is also reported. PMID- 16467988 TI - Molecular characterization and expression of a granzyme of an ectothermic vertebrate with chymase-like activity expressed in the cytotoxic cells of Nile tilapia (Oreochromis niloticus). AB - We have identified the gene coding for a novel serine protease with close similarities to mammalian granzymes from nonspecific cytotoxic cells of a teleost fish Oreochromis niloticus. The genomic organization of tilapia granzyme-1 (TLGR 1) has the signature five-exon-four-intron structure shared by all granzymes and similar hematopoietic Ser proteases. Molecular modeling studies suggested a granzyme-like structure for this protein with four disulfide linkages and two additional Cys residues. The expression of this gene is found to be restricted to cytotoxic cell populations with a low level of constitutive expression when compared to similar granzymes in other teleost species. High levels of transcriptional activation of TLGR-1 with different stimuli suggested that this gene is highly induced during immune reactions. Triplet residues around the active site Ser of TLGR, which determines the primary substrate specificity of granzymes, differ significantly from that of other granzymes. Recombinant TLGR-1 was expressed in the mature and proenzyme forms using pPICZ-alpha vector in the Pichia pastoris expression system. Recombinant TLGR-1 was used to determine the primary substrate specificity of this protease using various synthetic thiobenzyl ester substrates. In vitro enzyme kinetics assays suggested a preference for residues with bulky side chains at the P1 site, indicating a chymase-like activity for this protease. These results indicate the presence of novel granzymes in cytotoxic cells from ectothermic vertebrates. PMID- 16467990 TI - Growth response and modifications of organic nitrogen compounds in pure and mixed cultures of lactic acid bacteria from wine. AB - The interactions between the proteolytic X2L strain of Oenococcus oeni and the non-proteolytic 12p strain of Pediococcus pentosaceus were assayed. The characteristics of cell growth, protein degradation, and amino acid production of both strains were determined in pure and mixed cultures. O. oeni showed poor cell growth and greater ability in the release of amino acids to the extracellular medium, whereas P. pentosaceus showed a higher yield in cell production with a decrease in the amino acid concentration in the medium. P. pentosaceus especially consumed essential amino acids for growth, and O. oeni released several of the essential amino acids important for growth of P. pentosaceus. In the mixed culture, mutualism was observed. The higher activity of the proteolytic system of O. oeni in mixed culture produced an increase in cell growth and in the amount of essential amino acids released. These findings provide new knowledge about the metabolic interactions between lactic acid bacteria isolated from wine when proteins are degraded in mixed bacterial populations. PMID- 16467991 TI - Real-time PCR detection of the effects of protozoa on rumen bacteria in cattle. AB - A real-time PCR approach was used in this study to clarify the populations of major bacterial species in the rumens of faunated and unfaunated cattle. The sensitivity of this novel real-time PCR assay was evaluated by using 10(1) to 10(8) plasmid copies of target bacteria. The numbers of plasmid copies of Ruminococcus albus, Ruminococcus flavefaciens, Prevotella ruminicola, and the CUR E cluster were higher in the unfaunated than in the faunated rumens. The CUR-E cluster belongs to the Clostridium group. In contrast, Fibrobacter succinogenes was higher in the faunated than in the unfaunated rumens. Although it is well known that an absence of protozoa brings about an increase in the bacterial population, it was clarified here that an absence of protozoa exerted differential effects on the populations of cellulolytic bacteria in cattle rumens (i.e., F. succinogenes, R. albus, and R. flavefaciens). In addition, real-time PCR analysis suggested that the CUR-E cluster was more prevalent in the unfaunated rumens. PMID- 16467992 TI - Pythium rhizo-oryzae sp. nov. isolated from paddy fields: taxonomy, ITS region of rDNA, and comparison with related species. AB - Pythium rhizo-oryzae sp. nov. was isolated from some soil samples taken in paddy fields in northeastern India. For quite a long time it was wrongly identified as Pythium catenulatum Mathews, because of the presence of numerous catenulate hyphal bodies. The sequencing of the ITS region of its nuclear ribosomal DNA and a close look at its morphological characters have now enabled us to describe it as a new species. This species is characterized by the absence of zoospores and sporangia, rarely formed aritheridia and oogonia, and the presence of chained and terminal hyphal bodies. The ITS region of its rDNA comprises 774 bases. This oomycete is closely related to the members producing a filamentous inflated type of sporangia like P. catenulatum Mathews, P. torulosum Coker & Paterson, and P. inflatum Mathews. The closest of all these is P. catenulatum. Taxonomic description of this new species and its comparison with related oomycetes, together with the sequence of the PCR amplified internal transcribed region (spacers ITS1, ITS2, and the gene 5.8S) of its rDNA are given here. PMID- 16467993 TI - The mode of promoting activity of O-ethylmenthol as a transdermal absorption enhancer. AB - PURPOSE: The mode of action of O-ethylmenthol (MET), a promising compound to enhance transdermal drug delivery, was elucidated. Morphology of the skin treated with MET was investigated employing a laser scanning confocal microscopy. METHODS: Confocal scanning laser microscope and laser scanning microscope were employed for the morphological evaluation of the stratum corneum. To evaluate the fluidity of intercellular lipids by treatment with MET, liposomes composed of the stratum corneum lipids were prepared. RESULTS: Distribution amounts of the fluorescent probes greatly increased in the intercellular regions of the stratum corneum treated with 40% ethanol containing MET. Based on the skin surface observations, the difference in relative height between keratinocytes and intercellular regions was defined as DeltaH = DeltaH(keratinocytes) - DeltaH(intercellular space), where DeltaH is the difference in relative height, DeltaH(keratinocytes) is the height of center region in the keratinocytes, and DeltaH(intercellular space) is the height of the intercellular space. DeltaH values became negative in the skin surface treated with 40% ethanol containing MET because of the swelling in the intercellular regions. DeltaH values changed from positive to negative 15-30 min after the administration of MET. A very short period of application of MET was sufficient to induce its promoting activity. CONCLUSIONS: MET was able to change the structure of the intercellular lipids, thereby enhancing both the partitioning and diffusion of drugs through the skin. PMID- 16467995 TI - Urinary type I collagen cross-linked N-telopeptides in inhabitants 18 years after cessation of exposure to cadmium in Japan. PMID- 16467996 TI - Cancer health risk assessment of exposure to arsenic by workers of AngloGold Ashanti-Obuasi gold mine. PMID- 16467997 TI - Effects of environmentally relevant doses of cyanide on flight times in pigeons, Columba livia. PMID- 16467998 TI - Comparative toxicity of ammonium and perchlorate to amphibians. PMID- 16467999 TI - Acute and chronic toxicity of alcohol ethoxylates to the green alga, Desmodesmus (=Scenedesmus) subspicatus, and the subsequent development of structure activity relationships. PMID- 16468000 TI - Effects of chlorine disinfection on toxicity formation in reclaimed water. PMID- 16468001 TI - Biomarkers detected in Chub (Leuciscus cephalus L.) to evaluate contamination of the Elbe and Vltava rivers, Czech Republic. PMID- 16468002 TI - Endosulfan-induced genotoxicity detected in the Gilthead Seabream, Sparus aurata L., by means of flow cytometry and micronuclei assays. PMID- 16468003 TI - Effects of the fungicide mancozeb on liver structure of Nile tilapia, Oreochromis niloticus: assessment and quantification of induced cytological changes using qualitative histopathology and the stereological point-sampled intercept method. PMID- 16468004 TI - Recovery of Lemna sp. after exposure to sulfonylurea herbicides. PMID- 16468005 TI - Zinc tolerance and hyperaccumulation of Sedum alfredii Hance: a greenhouse experiment with artificial polluted soils. PMID- 16468006 TI - Spider webs as indicators of heavy metal pollution in air. PMID- 16468007 TI - Cadmium concentrations in blood in a group of male recruits in Slovenia related to smoking habits. PMID- 16468009 TI - Spatial and temporal variability of copper, zinc, and cobalt in marine macroalgae from the southwest coast of India. PMID- 16468008 TI - Effect of ecological variation on heavy metal content of some medicinal plants used as herbal tea ingredients in India. PMID- 16468010 TI - Contamination of toxic volatile and semi-volatile compounds from automobile materials. PMID- 16468011 TI - Particle size distribution and respiratory deposition estimates of airborne perfluorooctanoate and perfluorooctanesulfonate in Kyoto area, Japan. PMID- 16468012 TI - Statistical evaluation of the bioavailability of heavy metals from contaminated soils to vegetables. PMID- 16468013 TI - Atmospheric deposition of heavy metals in Thrace studied by analysis of Austrian pine (Pinus nigra) needles. PMID- 16468014 TI - Trace metal concentrations in different primary producers from Altata-Ensenada del Pabellon and Guaymas Bay (Gulf of California). PMID- 16468015 TI - Sorption of fipronil in tropical soils. PMID- 16468016 TI - Trichloroethylene contamination in fractured bedrock aquifer in Wonju, South Korea. PMID- 16468017 TI - Organochlorine pesticides and polychlorinated biphenyls in fish and fish products consumed by the population of the Republic of Belarus. PMID- 16468018 TI - Persistence of Acetamiprid in/on Mustard (Brassica juncea L.). PMID- 16468019 TI - Multiresidue analysis of pesticides in soil by gas chromatography with electron capture detection and gas chromatography mass spectrometry detection. PMID- 16468020 TI - The distribution pattern of pathology and cholinergic deficits in amygdaloid complex in Alzheimer's disease and dementia with Lewy bodies. AB - We studied the distribution pattern of pathology and cholinergic deficits in the subnuclei of the amygdaloid complex (AC) in five patients with Alzheimer's disease (AD), eight with dementia with Lewy bodies (DLB) and five normal controls. In controls, the basal nucleus contained the highest choline acetyltransferase activity; the activity in the lateral and central nuclei and those in the cortical, medial and accessory basal nuclei were comparable. In AD, there was a significant decrease in choline acetyltransferase activity in the accessory basal and lateral nuclei, in DLB a significant decrease was observed in the accessory basal, lateral and cortical nuclei. Compared to controls the hyperphosphorylated tau-pathology burden was significantly higher in the basal, central and medial nuclei in AD and in the central, cortical, lateral and medial nuclei in DLB. The amyloid plaque burden was significantly higher in the accessory basal, basal, lateral and cortical nuclei in AD and in all nuclei in DLB. The alpha-synuclein burden was significantly higher in all nuclei in both AD and DLB. Compared to AD alpha-synuclein burden was higher in all nuclei in DLB. There were no correlations between the distribution pattern of hyperphosphorylated tau-pathology, amyloid plaques and alpha-synuclein-positive structures, and choline acetyltransferase activity, except the lateral nucleus in DLB. In conclusion we found no relationship between the pattern of cholinergic deficits and the distribution pattern of lesions in the AC of patients with AD or DLB. Cholinergic deficits were more prominent in the nuclei of basolateral (BL) group in AD, whereas the nuclei of both BL and corticomedial groups were involved in DLB, which may be due to the involvement of both basal forebrain and brainstem cholinergic nuclei in the latter. PMID- 16468021 TI - Dysregulation of mitochondrial fusion and fission: an emerging concept in neurodegeneration. AB - Mitochondrial dysfunction is increasingly being recognized as an important factor contributing to the pathogenesis of neurodegenerative disorders. However, at present, the molecular basis underlying the decline in mitochondrial function is not really understood, but recent experimental evidence has shed some light on the pivotal role of mitochondrial morphology control in this process. In particular, dysregulated mitochondrial fusion and fission events can now be regarded as playing important pathogenic roles in neurodegeneration. In healthy cells, mitochondrial morphology is maintained through a dynamic balance between fusion and fission processes, and this regulated balance seems to be required for maintaining normal mitochondrial and cellular function. Moreover, during programmed cell death, activation of mitochondrial fission occurs, leading to mitochondrial fragmentation (Karbowski et al. in J Cell Biol 164:493-499, 2004). Consequently, inhibition of mitochondrial fission results in a significantly reduced cellular susceptibility toward apoptosis. The clinical relevance of maintaining a finely tuned balance between mitochondrial fusion and fission processes is underscored by the fact that the pathogenesis of certain hereditary neurodegenerative disorders such as autosomal dominant optic atrophy (ADOA) and Charcot-Marie-Tooth neuropathy type 2A (CMT2A) can now be linked to mutations in genes encoding mediators of mitochondrial fusion. In this article, I will summarize important aspects of what is currently known about the molecular machinery regulating mitochondrial fission and fusion in mammalian cells. Special emphasis will be given to the consequences of dysregulated mitochondrial morphology with regard to the pathogenesis of neurodegenerative disorders. A detailed understanding of the mitochondrial fission and fusion machinery will be a prerequisite for the development of therapeutic approaches to inhibit the neuronal cell death underlying certain neurodegenerative disorders. PMID- 16468022 TI - Interference with the renin-angiotensin system in heart failure. PMID- 16468024 TI - Development of an on-line automated sample clean-up method and liquid chromatography-tandem mass spectrometry analysis: application in an in vitro proteolytic assay. AB - Fluorescence detection has been a method of choice in industry for screening assays, including identification of enzyme inhibitors, owing to its high throughput capabilities, excellent reproducibility, and sensitivity. Occasionally, inhibitors are identified that challenge the fluorescence assay limit, necessitating the development of more sensitive detection methods to assess these compounds. For data mining purposes, however, original assay conditions may be required. A direct method transfer to highly sensitive and specific LC-MS-based methods has not always been possible due to the presence of MS-incompatible neutral detergents and non-volatile salts in the assay matrix. Utilizing an in vitro proteolytic screening assay for the serine protease hepatitis C virus (HCV) nonstructural (NS) 3 protease as a test case, we report the development of an automated sample clean-up procedure implemented on-line with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis to complement fluorescence detection. Ion exchange and peptide microtraps were employed to remove MS-incompatible assay matrix components. Three protease inhibitors were used to validate the MS/MS method. Comparable potencies were achieved for these compounds when assessed by fluorescence and MS/MS detection. Furthermore, four-fold less enzyme could be utilized when employing the MS/MS method compared to fluorescence detection. The longer analysis time, however, resulted in reduced sample capacity. The potency of our designed HCV NS3 protease inhibitors are thus routinely evaluated using a continuous fluorescence-based assay. Only pertinent inhibitors approaching the fluorescence assay sensitivity limit are subsequently analyzed further by LC-MS/MS. This methodology allows us to maintain a database and to compare results independent of the detection method. Despite the relatively slow sample turnaround time of this LC-MS approach, the versatility of the automated on-line clean-up procedure and sample analysis can be applied to assays containing reagents which were historically considered to be MS incompatible. PMID- 16468023 TI - A hAT superfamily transposase recruited by the cereal grass genome. AB - Transposable elements are ubiquitous genomic parasites with an ancient history of coexistence with their hosts. A few cases have emerged recently where these genetic elements have been recruited for normal function in the host organism. We have identified an expressed hobo/Ac/Tam (hAT) family transposase-like gene in cereal grasses which appears to represent such a case. This gene, which we have called gary, is found in one or two copies in barley, two diverged copies in rice and two very similar copies in hexaploid wheat. No gary homologues are found in Arabidopsis. In all three cereal species, an apparently complete 2.5 kb transposase-like open reading frame is present and nucleotide substitution data show evidence for positive selection, yet the predicted gary protein is probably not an active transposase, as judged by the absence of key amino acids required for transposase function. Gary is expressed in wheat and barley spikes and gary cDNA sequences are also found in rice, oat, rye, maize, sorghum and sugarcane. The short inverted terminal repeats, flanked by an eight-nucleotide host sequence duplication, which are characteristic of a hAT transposon are absent. Genetic mapping in barley shows that gary is located on the distal end of the long arm of chromosome 2H. Wheat homologues of gary map to the same approximate location on the wheat group 2 chromosomes by physical bin-mapping and the more closely related of the two rice garys maps to the syntenic location near the bottom of rice chromosome 4. These data suggest that gary has resided in a single genomic location for at least 60 Myr and has lost the ability to transpose, yet expresses a transposase-related protein that is being conserved under host selection. We propose that the gary transposase-like gene has been recruited by the cereal grasses for an unknown function. PMID- 16468026 TI - The haptic reproduction of orientations in three-dimensional space. AB - This research studied the haptic perception of orientations in space rather than in a plane. It aimed at identifying the nature of the system of coordinate used to represent an orientation in space, when two parameters are necessary to code an orientation. Blindfolded participants inserted the tip of the index finger in a thimble mounted at the extremity of a haptic interface, explored the orientation of a "virtual rod" with to-and-fro movements and, after a short delay, reproduced the same orientation with the same fingertip in the absence of the virtual rod. Globally, the haptic reproduction of orientations was anisotropic. When the reproduction of orientations was carried out in the frontal plane, a classical oblique effect (lower performance for the diagonal orientations than for the vertical and horizontal orientations) occurred. When the reproduction of orientations was carried out in space, orientations seemed to be coded in a coordinate system based on the sagittal plane. PMID- 16468025 TI - Remediating polluted soils. AB - This review focuses on treatment-based remediation of soils and the acquisition of data to support and monitor this remediation. Only in the last two decades has significant progress been made in regulating for soil pollution, with a parallel development of methodologies for soil assessment and remediation. However, soil complexity remains a problem for pollutant measurements relevant to environmental risk and informative to the design or evaluation of remediation technologies. Understanding the distribution of pollutants between different soil phases and the kinetics of transfer between these pools is fundamental to prediction for these processes; further progress is needed to characterise less accessible pollutant pools and to develop guidelines for their analysis. Available remediation options include physical, chemical and biological treatments, and these options offer potential technical solutions to most soil pollution. However, selecting the most appropriate approach requires detailed information on how pollutants interact with soil physio-chemical properties. Only general information is available as to the effectiveness of specific treatment systems for particular soil type-pollutant combinations. Given the high degree of heterogeneity in physio-chemical characteristics and pollutant distribution of affected soils, prediction of treatment timescales and levels of residual contamination remains a problem. On sites with a range of organic and inorganic pollutants present, combinations of different treatment approaches may offer the best prospect for effective remediation. Further work is needed to provide evidence that residual contamination does not pose significant risk and to evaluate effects of treatments on general soil function in relation to this contamination. PMID- 16468027 TI - A case in a child of giant left-atrial myxoma associated with recurrent high fever and myxoma cells expressing interleukin-6. PMID- 16468029 TI - Strangulation of internal hemorrhoids complicating sclerosing therapy with injection of OC-108 (Zione). PMID- 16468030 TI - A commentary on the article "Prevalence of the mismatch repair-deficient phenotype in colonic adenomas arising in HNPCC patients--results of a 5-year follow-up study". PMID- 16468031 TI - Risk of mistaken DNR orders. AB - A questionnaire study was carried out among attendants at a community cancer center to determine the subjects' preferences and understanding of the meaning of do-not-resuscitate (DNR). Only 34% correctly understood the meaning of DNR, and 66% thought that DNR was administered only to prolong life without realizing that a DNR decision would result in not being resuscitated even if the cause of the sudden death was potentially reversible. We then determined the subjects' preferences if they had developed a treatment complication needing resuscitation and be put on the ventilator machine temporarily. When the subject was not expected to be alive in 6 months, the preference for resuscitation was not related to correct understanding. However, when the chance of cure was 30%, a preference for resuscitation was related to an incorrect understanding of the meaning of DNR. About 70% of respondents who would accept ventilator care had an incorrect understanding of DNR. The adjusted odds for the correct understanding of DNR were less for respondents who preferred resuscitation. The adjusted odds ratio (AOR) was 0.58 (CI: 0.35-0.93) (p=0.02) after adjusting for age and 0.53 (CI: 0.32-0.86) (p=0.01) after adjusting for both age and treatment group. These results suggest that physicians should be open to the possibility that patients may not always understand what DNR means, and they may be placed on DNR by mistake. PMID- 16468033 TI - Hydrogen-bonding conformations of tyrosine B10 tailor the hemeprotein reactivity of ferryl species. AB - Ferryl compounds [Fe(IV)=O] in living organisms play an essential role in the radical catalytic cycle and degradation processes of hemeproteins. We studied the reactions between H2O2 and hemoglobin II (HbII) (GlnE7, TyrB10, PheCD1, PheE11), recombinant hemoglobin I (HbI) (GlnE7, PheB10, PheCD1, PheE11), and the HbI PheB10Tyr mutant of L. pectinata. We found that the tyrosine residue in the B10 position tailors, in two very distinct ways, the reactivity of the ferryl species, compounds I and II. First, increasing the reaction pH from 4.86 to 7.50, and then to 11.2, caused the the second-order rate constant for HbII to decrease from 141.60 to 77.78 M-1 s-1, and to 2.96 M-1 s-1, respectively. This pH dependence is associated with the disruption of the heme-tyrosine (603 nm) protein moiety, which controls the access of the H2O2 to the hemeprotein active center, thus regulating the formation of the ferryl species. Second, the presence of compound I was evident in the UV-vis spectra (648-nm band) in the reactions of HbI and recombinant HbI with H2O2, This band, however, is completely absent in the analogous reaction with HbII and the HbI PheB10Tyr mutant. Therefore, the existence of a hydrogen-bonding network between the heme pocket amino acids (i.e., TyrB10) and the ferryl compound I created a path much faster than 3.0x10( 2) s-1 for the decay of compound I to compound II. Furthermore, the decay of the heme ferryl compound I to compound II was independent of the proximal HisF8 trans ligand strength. Thus, the pH dependence of the heme-tyrosine moiety complex determined the overall reaction rate of the oxidative reaction limiting the interaction with H2O2 at neutral pH. The hydrogen-bonding strength between the TyrB10 and the heme ferryl species suggests the presence of a cycle where the ferryl consumption by the ferric heme increases significantly the pseudoperoxidase activity of these hemeproteins. PMID- 16468032 TI - Diethylstilbestrol increases the density of prolactin cells in male mouse pituitary by inducing proliferation of prolactin cells and transdifferentiation of gonadotropic cells. AB - Diethylstilbestrol (DES) has been implicated in mammalian abnormalities. We examined the effects of DES on follicle-stimulating hormone (FSH), luteinizing hormone (LH), and prolactin (PRL) cells in the pituitaries of male mice treated with various doses of DES for 20 days. DES reduced the density of FSH and LH cells in a dose-dependent manner, but increased that of PRL cells. When the expression of estrogen receptor (ER) alpha and beta was assessed, an induction of ERbeta by DES was found predominantly in PRL cells. However, since these effects were abolished in ERalpha knockout mice, DES appears to act primarily through ERalpha. When the expression of Ki-67 and Pit-1 in PRL cells was examined at various time-points after DES treatment, some PRL cells became Ki-67 positive at 10-15 days, and Pit-1-positive cells were increased at 5-15 days. Furthermore, some FSH and LH cells became Pit-1 positive, and co-localized with PRL at 5-10 days. Our results indicate that DES increases PRL cells by inducing proliferation of PRL cells and transdifferentiation of FSH/LH cells to PRL cells. PMID- 16468034 TI - In vitro priming of tumor-specific cytotoxic T lymphocytes using allogeneic dendritic cells derived from the human MUTZ-3 cell line. AB - The adoptive transfer of in vitro-induced and expanded tumor-specific cytotoxic T lymphocytes (CTL) presents a promising immunotherapeutic approach for the treatment of cancer. The in vitro induction of tumor-reactive CTL requires repeated stimulation of CTL precursors with dendritic cells (DC). To circumvent problems like scarcity of blood DC precursors and donor variability, it would be attractive to use DC from a non-autologous, unlimited source. DCs derived from the human acute myeloid leukemia (AML) cell line MUTZ-3 are attractive candidates since these DCs closely resemble monocyte-derived DC (MoDC) in terms of phenotype and T cell stimulatory capacity. Here we demonstrate that functional CTL clones could be generated against multiple tumor-associated antigens, i.e., human telomerase reverse transcriptase (hTERT), ErbB3-binding protein-1 (Ebp1), carcinoembryonic antigen (CEA) and Her-2/neu, by stimulating CD8beta(+) CTL precursors with peptide-loaded allogeneic, HLA-A2-matched MUTZ-3-derived DC. A consistent induction capacity, as determined by MHC tetramer-binding, was found in multiple donors and comparable to autologous peptide-loaded MoDC. Functional characterization at the clonal level revealed the priming of CTL that recognized endogenously processed epitopes on tumor cell lines in an HLA-A2-restricted fashion. Our data indicate that MUTZ-3-derived DC can be used as stimulator cells for in vitro priming and expansion of functional TAA-specific effector CTL. MUTZ 3-derived DCs thus represent a ready and standardized source of allogeneic DC to generate CTL for therapeutic adoptive transfer strategies. PMID- 16468036 TI - [Does anesthesia damage children's brains?]. PMID- 16468035 TI - Tumor progression despite massive influx of activated CD8(+) T cells in a patient with malignant melanoma ascites. AB - Although melanoma tumors usually express antigens that can be recognized by T cells, immune-mediated tumor rejection is rare. In many cases this is despite the presence of high frequencies of circulating tumor antigen-specific T cells, suggesting that tumor resistance downstream from T cell priming represents a critical barrier. Analyzing T cells directly from the melanoma tumor microenvironment, as well as the nature of the microenvironment itself, is central for understanding the key downstream mechanisms of tumor escape. In the current report we have studied tumor-associated lymphocytes from a patient with metastatic melanoma and large volume malignant ascites. The ascites fluid showed abundant tumor cells that expressed common melanoma antigens and retained expression of class I MHC and antigen processing machinery. The ascites fluid contained the chemokines CCL10, CCL15, and CCL18 which was associated with a large influx of activated T cells, including CD8(+) T cells recognizing HLA-A2 tetramer complexes with peptides from Melan-A and NA17-A. However, several functional defects of these tumor antigen-specific T cells were seen, including poor production of IFN-gamma in response to peptide-pulsed APC or autologous tumor cells, and lack of expression of perforin. Although these defects were T cell intrinsic, we also observed abundant CD4(+)CD25(+)FoxP3(+) T cells, as well as transcripts for FoxP3, IL-10, PD-L1/B7-H1, and indoleamine-2,3-dioxygenase (IDO). Our observations suggest that, despite recruitment of large numbers of activated CD8(+) T cells into the tumor microenvironment, T cell hyporesponsiveness and additional negative regulatory mechanisms can limit the effector phase of the anti-tumor immune response. PMID- 16468037 TI - [Structural damage of epidural catheters during combined spinal epidural anaesthesia: a lightmicroscopical study]. AB - BACKGROUND: The purpose of this study was to investigate whether there is a risk of epidural catheter damage during the advancement of the spinal needle through an epidural needle in clinical use. METHODS: A total of 100 catheters (50 from CSE kits with a pencil-point type spinal needle and 50 from CSE kits with a Quincke type spinal needle) which had been used for routine CSE blocks were microscopically examined for any defects within the first 150 mm of the catheter. Additionally 10 unused new catheters were investigated. RESULTS: Among 10 unused catheters 5 slight scratches were found, 92 out of 100 used catheters did not show any signs of use or scratches, 7 showed some signs of use and longitudinal scratches whereas another 1 showed a moderate scratch of less than 25% of the wall thickness. There was no difference in the prevalence of scratches between the CSE kits with pencil-point type spinal needles compared to those with Quincke type spinal needles. CONCLUSION: The CSE technique with either pencil-point type or Quincke-type spinal needles for subarachnoidal punctures was safe and showed no relevant epidural catheter damage. PMID- 16468038 TI - Genotype of galectin 2 (LGALS2) is associated with insulin-glucose profile in the British Women's Heart and Health Study. AB - AIMS/HYPOTHESIS: It has been suggested that the gene encoding lymphotoxin-alpha (LTA) is associated with insulin resistance, and genetic association studies in the LTA region offer some support for this. However, LTA is in linkage disequilibrium with both the HLA gene cluster and the gene encoding TNF-alpha, making inferences about causality difficult. In this study, we used the galectin 2 (LGALS2) genotype, which affects LTA secretion but is located on another chromosome than the HLA gene cluster or TNF, to examine the relationship between the LTA pathway and traits of the metabolic syndrome. SUBJECTS: A cross-sectional genetic association study was carried out in 3,272 British women of European origin who were aged 60 to 79 years and were randomly selected from the community. RESULTS: Fasting plasma glucose and serum insulin were statistically significantly associated with LGALS2 rs7291467, with this association being independent of BMI and WHR. The mean difference in fasting insulin per minor allele was -4% (p=0.01 for trend by allele) and the mean per minor allele difference in fasting glucose was -2% (p=0.02 for trend by allele). When women with known diabetes were excluded from the analyses the findings did not differ from those for the whole cohort. CONCLUSIONS/INTERPRETATION: Our findings for the physically unlinked LGALS2, invite further study of LGALS2 specifically and the LTA pathway generally for their influence on glucose-insulin regulation. PMID- 16468039 TI - Gene expression analysis in mice liver on hepatocarcinogenesis by flumequine. AB - mRNA expression profiles in the liver from mice treated with flumequine (FL) were analyzed in order to elucidate the mechanism of its tumor-promoting effect. The liver from a C3H/He mouse that received a diet containing 4,000 ppm of FL for 4 weeks was examined by cDNA microarray in comparison with an untreated mouse. Furthermore, to obtain a more comprehensive sequence, time-course changes in selected genes were determined by real-time RT-PCR. Microarray analysis revealed 15 upregulated and 9 downregulated genes in an FL-treated mouse. The upregulated genes included signal transducers and cell cycle regulators. In addition, the levels of stress response genes, particularly glutathione S-transferase (GST) alpha and GSTmu, were very high, indicating the generation of oxidative stress. On the other hand, the downregulated genes included phase I metabolic enzymes, such as cytochrome P450 (CYPs) enzymes, and apoptosis-associated proteins. These changes were confirmed by quantitative RT-PCR and were generally consistent with each other. Time-course observations revealed consistent results, particularly with regard to GSTalpha, GSTmu, ERK5, and CYP2E1. In addition, the expression of 8-oxoguanine DNA glycosylase 1 (OGG1) was increased in a time-dependent manner. These results suggest the possibility that responses against oxidative stress may play a major role in hepatocarcinogenesis by FL in mice. PMID- 16468040 TI - Agrobacterium T-DNA-mediated integration and gene replacement in the brown rot pathogen Monilinia fructicola. AB - A transformation system utilizing Agrobacterium tumefaciens was developed for targeted gene disruption in Monilinia fructicola, a fungal pathogen that causes brown rot disease of stone fruits. Transformation with a vector containing the neomycin phosphotransferase II (nptII) cassette flanked with 4 kb cutinase gene (Mfcut1) flanking sequences resulted in an average of 13 transformants per 10(5) spores. When assayed by PCR and DNA blot analyses, more than 50% of the transformants recovered had integrated in the targeted Mfcut1 locus. Both target gene-specific and non-specific integrations carried direct (head-to-tail) repeat T-DNA integrations. Sequence analysis of these T-DNA integrations revealed that 26 bp of the T-DNA right border were missing at the junctions between direct repeats in all cases. The recombination event during non-specific T-DNA integration in this fungus was unlike that reported in Agrobacterium-mediated transformation in plants. PMID- 16468041 TI - Comparative analysis of HOG pathway proteins to generate hypotheses for functional analysis. AB - Comparative genomics allows comparison of different proteins that execute presumably identical functions in different organisms. In contrast to paralogues, orthologues per definition perform the same function and interact with the same partners and, consequently, should display conservation in all these properties. We have employed 20 fungal genomes to analyse key components of the high osmolarity glycerol signalling pathway of Saccharomyces cerevisiae. Among the proteins scrutinised are a complete phosphotransfer module, a MAP kinase, two scaffold proteins, one of which is also a MAPKK, and two transcription factors. Sequence alignments, domain structure and size analysis, combined with the rich information available in the literature, allowed us to probe previous structural and functional studies and to generate hypotheses for future experimental studies. Although certain domains are too highly conserved across fungal species for meaningful comparative studies, others, like interaction domains, can be studied in closely related species. Moreover, putative functionally relevant sites for protein modifications can be identified in such comparative studies. We provide several relevant examples and present a number of previously un(der)characterised domains of potential functional significance in osmosensing and signal transduction. We propose that any functional protein analysis in fungi should make use of the unique resource that fungal genome sequences offer. PMID- 16468042 TI - Comparative genomics of the HOG-signalling system in fungi. AB - Signal transduction pathways play crucial roles in cellular adaptation to environmental changes. In this study, we employed comparative genomics to analyse the high osmolarity glycerol pathway in fungi. This system contains several signalling modules that are used throughout eukaryotic evolution, such as a mitogen-activated protein kinase and a phosphorelay module. Here we describe the identification of pathway components in 20 fungal species. Although certain proteins proved difficult to identify due to low sequence conservation, a main limitation was incomplete, low coverage genomic sequences and fragmentary genome annotation. Still, the pathway was readily reconstructed in each species, and its architecture could be compared. The most striking difference concerned the Sho1 branch, which frequently does not appear to activate the Hog1 MAPK module, although its components are conserved in all but one species. In addition, two species lacked apparent orthologues for the Sln1 osmosensing histidine kinase. All information gathered has been compiled in an MS Excel sheet, which also contains interactive visualisation tools. In addition to primary sequence analysis, we employed analysis of protein size conservation. Protein size appears to be conserved largely independently from primary sequence and thus provides an additional tool for functional analysis and orthologue identification. PMID- 16468043 TI - Enhanced production of MMP-1, MMP-3, MMP-13, and RANTES by interaction of chondrocytes with autologous T cells. AB - It has been reported that T cells and chondrocytes interact through cell surface molecules such as MHC, CD4 or CD8 in osteoarthritis (OA) and T cells are activated. The objective of this study is to investigate the responses of chondrocyte-T cell interaction in terms of metalloprotease (MMP) and chemokine production. Articular cartilage and autologous blood were obtained from patients with OA and fracture who under went prosthetic surgery. Synovial fluid (SF) was collected from OA patients. Isolated chondrocytes were co-cultured with autologous T cells. SF cells were analyzed by immunostaining or Alcian blue staining. The production of MMP-1, MMP-3, MMP-13, and regulated on activation, normal T expressed and secreted (RANTES) was enhanced by direct co-culture compared to indirect co-culture using Transwell. Production ratio of RANTES in OA was significantly higher than non-arthritic samples. CD3 positive mononuclear cells and chondrocyte-like cells were found in SF. Chondrocyte-T cell contact was more adhesive in OA samples. These results showed the production of MMPs and RANTES was enhanced by the interaction and that chondrocyte-T cell contact was possible in vivo. PMID- 16468044 TI - Role of mitogen-activated protein kinases and NFkappaB on IL-1beta-induced effects on collagen type II, MMP-1 and 13 mRNA expression in normal articular human chondrocytes. AB - Interleukin-1ss is a pro-inflammatory cytokine that causes anti-anabolic and catabolic effects on articular chondrocytes via four major signaling pathways. In this study, we investigated the role of these pathways for the repression of collagen type II, and induction of MMP-1 and -13 by Il-1ss. Human adult chondrocytes were stimulated with IL-1beta together with selective inhibitors of the ERK, JNK, p38, and NFkappaB pathways. Inhibitors of ERK and NFkappaB could significantly block the induction of MMP-1 and -13 (p<0.05) and the repression of collagen type II (p<0.01). The inhibitor for p38 MAPK was able to block partially MMP-1 and -13 up-regulation (p<0.01), but did not significantly inhibit collagen type II repression. Our data suggest that ERK and NFkB pathways are particularly important for IL-1beta regulating collagen type II and MMP-1 and -13 expression and that p38, but not JNK is additionally involved in MMP-1 and -13 induction. PMID- 16468046 TI - Examining the psychometric characteristics of the Dutch childhood health assessment questionnaire: room for improvement? AB - The aim of this study was to examine the psychometric characteristics of the childhood health assessment questionnaire-disability index (CHAQ-DI). Seventy-six patients with juvenile idiopathic arthritis (JIA), age range 4.8-15.8 years, completed a CHAQ questionnaire one or more times. In total, 321 CHAQ questionnaires were available for analysis. Factor analysis and correlation were used to analyse the data. The analysis indicated that 12 items could be removed from the original 30 items of the CHAQ-DI. Also the addition of "aids and assistance" to the overall scoring method of the CHAQ-DI did not contribute to the overall measuring concept of the CHAQ-DI. The psychometric characteristics of the CHAQ-DI could be improved by removing 12 items from the original 30 items. Moreover, a simple scoring method, without the addition of aids and assistance to the total CHAQ-DI improves sensitivity to change of the CHAQ-DI. PMID- 16468045 TI - Manifestation of rheumatoid arthritis in a patient with hereditary haemochromatosis. AB - Articular symptoms are frequent manifestations of hereditary haemochromatosis. The clinical signs of the arthropathy of haemochromatosis are not specific and difficult to identify in case of co-incidence of haemochromatosis with Heberden's and Bouchard's osteoarthritis or rheumatoid arthritis (RA). Here the manifestation of RA in a patient is reported who was successfully treated for haemochromatosis. Six months after terminating phlebotomy, the patient presented again suffering from impressive swelling of all MCP joints, showing strong synovitis in ultrasound, and from morning stiffness longer than 1 h. ESR, CRP, IgM rheumatoid factor, and anti-cyclic citrullinated peptide antibodies were markedly elevated. Based on these findings the diagnosis of RA was made. Therefore, the high prevalence of RA and haemochromatosis in the general population underlines the usefulness of a screening for HFE gene mutations in RA patients with an atypical course of the disease as well as in patients with undifferentiated arthritis. PMID- 16468047 TI - Transport of acetate and sodium in sheep omasum: mutual, but asymmetric interactions. AB - We have studied the transport of acetate across the isolated epithelium of sheep omasum; no net transport was observed (J(ms) approximately = J(sm)) under Ussing chamber conditions. Low mucosal pH (pH 6.4) significantly enhanced J(ms) acetate and the transport rates of acetate increased linearly and significantly (r2=0.99) with the luminal acetate concentration. The presence of another short chain fatty acid (propionate) did not affect J(ms) acetate significantly. Neither addition of 1 mmol l(-1) DIDS to the mucosal side nor HCO3 replacement caused changes of J(ms) acetate; this does not support the assumption of acetate transport via anion exchange. Addition of 1 mmol l(-1) amiloride to the mucosal side significantly decreased acetate fluxes at high mucosal acetate concentration (100 mmol l(-1)) and low pH (6.4) indicating interaction between acetate uptake in the undissociated form, intracellular release of protons and activation of Na+/H+ exchange (NHE). However, the mutual interaction between Na transport via NHE and acetate transport is asymmetric. Stimulation or inhibition of Na transport via NHE is much more pronounced than the corresponding changes of acetate fluxes. Thus, the obtained results support the conclusion that acetate is transported via simple diffusion and probably predominantly in the protonated form, thereby explaining the positive and mutual interaction between Na transport and short chain fatty acids. PMID- 16468048 TI - Percutaneous resection of a patellar osteoid osteoma using a cannulated skin punch biopsy needle. AB - Patella is a very rare localization for osteoid osteoma. Non-specific knee complaints and difficulty to distinguish nidus in direct radiographs may cause a delay in diagnosis and make the definite diagnosis troublesome. The most effective and non-invasive method in treatment of osteoma is CT guided excision of the nidus. We present a case of patellar osteoid osteoma diagnosed by MRI scans. After being marked under CT guidance, the lesion was completely excised with a skin punch. We propose that, this method is both minimal invasive and effective in the management of patellar osteoid osteoma. PMID- 16468049 TI - Successful posterior interlaminar fusion at the thoracic spine by sole use of beta-tricalcium phosphate. AB - We report on a 43-year-old male who sustained an isolated distraction injury of the thoracic spine Th7/Th8 (AO/ASIF B 2.3) with wedge compression Th8 and sagittal split Th10 without neurological injury. A bisegmental posterior stabilisation and a monosegmental interlaminar fusion was the treatment of choice. A synthetic bone substitute, beta-tricalcium phosphate (beta-TCP, Chronos) without additional autogenous bone was used to achieve the monosegmental posterior fusion. The clinical course was favourable and 10 months postoperatively the implant was removed. On implant removal the CT scan showed a fused segment and intraoperatively it was found that the fusion was solidly healed. A biopsy was taken from the fusion mass and histology showed vital bone that was rich with osteocytes. Noncalcified osteoid surrounding the bone marrow cavity could be identified. Several studies and the reported case might indicate that osteoconductive material alone can be sufficient for achieving a solid fusion. PMID- 16468050 TI - Radial head prosthesis after fracture of radial head with associated elbow instability. AB - INTRODUCTION: Fractures of the radial head and associated elbow instability can be treated with operation with radial head prosthesis. In this study, we evaluate function 1-7 years after implantation and also function after removal of five prostheses. MATERIAL AND METHODS: Eighteen patients with radial head fracture and associated elbow instability were evaluated 3.7 years (1-7) after implantation of a radial head prosthesis. Pain at rest and during activity was measured with a visual analogue scale (VAS). Test of stability and neurological examination was done manually as well as measurement of the range of motion, using a goniometer. Activity of daily living (ADL) was estimated using five questions where the answers were graded between 1 and 3. The patients were asked to grade their general satisfaction according to the following scale; very satisfied, satisfied, not satisfied, disappointed. Plain X-rays were taken and 14 patients agreed to have their elbow strength evaluated using the validated BTE work simulator. RESULTS: Five prostheses had been extracted due to poor range of motion. All these patients improved after extraction. All elbows were stable. No patient with extracted prosthesis had VAS score >2. The mean extension defect for this group was 15 degrees (5-25) compared to the mean extension defect for the 13 patients with the prosthesis still in place 15 degrees (0-40). The highest VAS score for the patients with prosthesis was five but the mean as low as 0.8. In the whole group, 13 patients were pain free. ADL function was good in general. The X-rays of the prostheses, still in place, showed radiolucent lines in 7 of the 13 patients. In the whole group, there was a significant decrease in supination, flexion and extension strength (P<0.01, P<0.01, P<0.05). DISCUSSION: Radial head prosthesis works as a spacer after fracture of the radial head and associated instability. If range of motion is much restricted post-operatively, the prosthesis can be removed with improved function as result. PMID- 16468051 TI - Adenosine produced via the CD73/ecto-5'-nucleotidase pathway has no impact on erythropoietin production but is associated with reduced kidney weight. AB - CD73/ecto-5'-nucleotidase, which catalyzes the conversion of adenosine monophosphate to adenosine, has been implicated in vascular homeostasis. The aim of the present study was to evaluate the role of CD73 in erythropoietin (EPO) production and to determine its influence on basal kidney perfusion using a CD73 knockout mutant recently generated by us. Of all organs investigated, kidneys showed the most prominent CD73 activity, preferentially located in peritubular fibroblasts of the renal cortex and the glomerular mesangium. In the absence of CD73, alkaline phosphatase remained unchanged, but tissue adenosine was reduced under control conditions (by 76%) and during normobaric hypoxia (by 72%). Despite the loss of CD73 activity, EPO mRNA and plasma protein concentrations were not different under basal conditions as well as after normobaric hypoxia (8% O2) and carbon monoxide (0.1% CO) inhalation (both for 4 h). Although there were no differences in blood pressure and urine flow volume, average weight of both kidneys was reduced by 21% in the knockout (wild type 7.17+/-1.18 mg g-1 body wt, CD73-/- 5.70+/-1.91 mg g-1 body wt). Measurement of renal plasma flow and glomerular filtration revealed no significant differences when related to respective kidney weights. We conclude that adenosine derived by the extracellular CD73 pathway has no impact on EPO production under basal conditions and after hypoxic challenge but may determine kidney weight. PMID- 16468052 TI - Investigation of bioactive NO-scavenging role of myoglobin in myocardium. AB - Because nitric oxide (NO) can react with myoglobin (Mb) to oxidize the heme Fe(II) to Fe(III), the appearance of metmyoglobin (metMb) during bradykinin stimulation underpins the hypothesis that Mb acts as an NO scavenger in the cell. Although some experiments have detected the reporter metMb signal in the -3.7 ppm spectral region, others have not corroborated the finding. Because metMb also has characteristic hyperfine-shifted signals in the 40-100 ppm spectral region, detection of these signals would confirm the presence of metMb. Perfused rat myocardium study has examined this spectral region in a range of bradykinin infusion protocols. Although bradykinin elicits a set of physiological responses, consistent with the induction of NO, the (1)H nuclear magnetic resonance spectra in all experiments reveal no detectable metMb signals. Moreover, in the perfused myocardium model, the bradykinin-induced decline in myocardial oxygen consumption does not appear to arise only from NO binding to cytochrome oxidase. PMID- 16468053 TI - Evidence for the presence of thyroid stimulating hormone, thyroglobulin and their receptors in Eisenia fetida: a multilevel hormonal interface between the nervous system and the peripheral tissues. AB - The present study describes the localization and distribution of thyroid stimulating hormone (TSH), thyroglobulin (TGB) and their receptors in Eisenia fetida (Annelida, Oligochaeta) as revealed by immunohistological methods. Immunopositive neuronal and non-neuronal cells are present in both the central nervous system and some peripheral organs (e.g. foregut and coelomocytes). TSH- and TGB-immunopositive neurons in the various ganglia of the central nervous system are differentially distributed. Most of the immunoreactive cells are found in the suboesophageal ganglion. The stained cells also differ in their shapes (round, oval, pear-shaped) and sizes (small, 12-25 microm; medium, 20-35 microm; large, 30-50 microm). In all ganglia of the central nervous system, TSH-positive neurons additionally show gamma aminobutyric acid (GABA) immunopositivity. Non neuronal cells also take part in hormone secretion and transport. Elongated TSH positive cells have been detected in the capsule of the central ganglia and bear granules or vacuoles in areas lacking neurons. Many of capillaries show immunoreactivity for all four tested antibodies in the entire central nervous system and foregut. Among the coelomocytes, granulocytes and eleocytes stain for TSH and its receptor and for TGB but not for thyroid hormone receptor. Most of the granulocytes are large (25-50 microm) but a population of small cells (10-25 microm) are also immunoreactive. None of the coelomocytes stain for GABA. We therefore suggest that the members of this hormone system can modify both metabolism and immune functions in Eisenia. Coelomocytes might be able to secrete, transport and eliminate hormones in this system. PMID- 16468054 TI - Reproductive tactics under severe egg predation: an eider's dilemma. AB - Parental defence against predators may increase offspring survival but entail other costs. Egg predation is frequent early in the laying sequence of the common eider, which differs in this and in several other ways from most other waterfowl. We test the hypothesis that permanent presence at the nest from the second or third egg is an adaptation for reducing egg predation in eiders. Two other alternative hypotheses for lower predation at later nest stages are early predation loss of the most vulnerable nests and seasonal decrease in predation risk. Analyses of predation rates at the one-egg and later stages refute these two alternatives. Early nest attendance by eider females is estimated to increase clutch survival by about 20% in four-egg and 35% in five-egg clutches, albeit probably at a cost of smaller clutch size. PMID- 16468055 TI - Genotype and environment determine allocation to and costs of resistance in quaking aspen. AB - Although genetic variability and resource availability both influence plant chemical composition, little is known about how these factors interact to modulate costs of resistance, expressed as negative correlations between growth and defense. We evaluated genotype x environment effects on foliar chemistry and growth of quaking aspen (Populus tremuloides) by growing multiple aspen genotypes under variable conditions of light and soil nutrient availability in a common garden. Foliage was analyzed for levels of nitrogen, phenolic glycosides and condensed tannins. Bioassays of leaf quality were conducted with fourth-stadium gypsy moth (Lymantria dispar) larvae. Results revealed strong effects of plant genotype, light availability and nutrient availability; the importance of each factor depended upon compound type. For example, tannin concentrations differed little among genotypes and across nutrient regimes under low light conditions, but markedly so under high light conditions. Phenolic glycoside concentrations, in contrast, were largely determined by genotype. Variation in phenolic glycoside concentrations among genotypes was the most important factor affecting gypsy moth performance. Gypsy moth biomass and development time were negatively and positively correlated, respectively, with phenolic glycoside levels. Allocation to phenolic glycosides appeared to be costly in terms of growth, but only under resource-limiting conditions. Context-dependent trade-offs help to explain why costs of allocation to resistance are often difficult to demonstrate. PMID- 16468056 TI - Growth and leaf physiology of monkeyflowers with different altitude ranges. AB - Every species is limited both geographically and ecologically to a subset of available habitats, yet for many species the causes of distribution limits are unknown. Temperature is thought to be one of the primary determinants of species distributions along latitudinal and altitudinal gradients. This study examined leaf physiology and plant performance under contrasting temperature regimes of sister species of monkeyflower, Mimulus cardinalis and Mimulus lewisii (Phrymaceae), that differ in altitude distribution to test the hypothesis that temperature is the primary determinant of differences in fitness versus altitude. Each species attained greatest aboveground biomass, net photosynthetic rate, and effective quantum yield of photosystem II when grown under temperatures characteristic of the altitudinal range center. Although both species exhibited greater stem length, stomatal conductance, and intercellular CO2 concentration in hot than in cold temperatures, these traits showed much greater reductions under cold temperature for M. cardinalis than for M. lewisii. Survival of M. lewisii was also sensitive to temperature, showing a striking decrease in hot temperatures. Within each temperature regime, the species native to that temperature displayed greatest growth and leaf physiological capacity. Populations from the altitude range center and range margin of each species were used to examine population differentiation, but central and marginal populations did not differ in most growth or leaf physiological responses to temperature. This study provides evidence that M. cardinalis and M. lewisii differ in survival, growth, and leaf physiology under temperature regimes characterizing their contrasting low and high altitude range centers, and suggests that the species' altitude range limits may arise, in part, due to metabolic limitations on growth that ultimately decrease survival and limit reproduction. PMID- 16468057 TI - Prevention of latex allergy among health care workers and in the general population: latex protein content in devices commonly used in hospitals and general practice. AB - BACKGROUND: In this study the latex protein content in devices commonly used in hospitals and general practice were investigated. The main aim was to acquire information for preventing latex allergy in health care workers and in the general population. METHODS: About 22 different types of medical devices and 23 devices commonly used in general practice were examined evaluating the total allergenic potency by a modified RAST-inhibition assay and quantitative determination of single allergens (Hev b1, Hev b5 and Hev b6.02) by using commercial ELISA kit. RESULTS: A high level of inhibition was found in medical devices, such as elastic bandage (81.57%), tourniquet (74.09%), Foley urinary catheter (68.35%), Penrose drainage (67.25%) and taping (39.6%), and in common devices, such as rubber inner-sole (84.20%), toy balloon (78.62%), latex mattress (74.27%), household rubber gloves (49.10%), working gloves (38.25%), inflatable floating mattress (32.10%). Concentrations of latex extractable proteins and Hev b1, Hev b5 and Hev b6.02 antigens were high in some medical and general devices. CONCLUSIONS: Latex exposure sources were found in hospitals and the home. These findings, though only preliminary and far from conclusive, could enable sensitized persons to avoid risky exposures and prevent allergic reactions. From the point of view of prevention, the time may come when every natural rubber object could be systematically labelled as "containing latex" together with the warning that "this item may cause allergic reactions in sensitized subjects." PMID- 16468058 TI - Mechanically versus electro-magnetically braked cycle ergometer: performance and energy cost of the Wingate Anaerobic Test. AB - Performance and metabolic profiles of the Wingate Anaerobic Test (WAnT) were compared between a mechanically resisted (ME) and an electro-magnetically braked (EE) cycle ergometer. Fifteen healthy subjects (24.0+/-3.5 years, 180.5+/-6.1 cm, 75.4+/-11.9 kg) performed a WAnT on ME, and EE 3 days apart. Performance was measured as peak power (PP), minimum power (MP), mean power (AP), time to PP (TTPP), fatigue rate (FR), and maximum cadence (RPM(MAX)). Lactic (W (LAC)) and alactic (W (PCR)) anaerobic energy were calculated from net lactate appearance and the fast component of post-exercise oxygen uptake. Aerobic metabolism (W (AER)) was calculated from oxygen uptake during the WAnT. Total energy cost (W (TOT)) was calculated as the sum of W (LAC), W (PCR), and W (AER). There was no difference between ME and EE in PP (873+/-159 vs. 931+/-193 W) or AP (633+/-89 vs. 630+/-89 W). In the EE condition TTPP (2.3+/-0.7 vs. 4.3+/-0.7 s) was longer (P<0.001), MP (464+/-78 vs. 388+/-57 W) was lower (P<0.001), FR (15.2+/-5.2 vs. 20.5+/-6.8%) was higher (P<0.005), and RPM(MAX) (168+/-18 vs. 128+/-15 rpm) was slower (P<0.001). There was no difference in W (TOT) (1,331+/-182 vs. 1,373+/-120 J kg(-1)), W (AER) (292+/-76 vs. 309+/-72 J kg(-1)), W (PCR) (495+/-153 vs. 515+/ 111 J kg(-1)) or W (LAC) (545+/-132 vs. 549+/-141 J kg(-1)) between ME and EE devices. The EE produces distinctly different performance measures but valid metabolic WAnT results that may be used to evaluate anaerobic fitness. PMID- 16468060 TI - RAAS polymorphisms alter the acute blood pressure response to aerobic exercise among men with hypertension. AB - Limited evidence suggests renin-angiotensin-aldosterone system (RAAS) polymorphisms alter the blood pressure (BP) response to aerobic exercise training. We examined if RAAS polymorphisms influenced postexercise hypotension in men with high normal to Stage 1 hypertension. Forty-seven men (44.2+/-1.4 years, 145.1+/-1.6/85.5+/-1.1 mmHg) randomly completed three experiments: seated rest (control) and two cycle exercise bouts at 40% (LITE) and 60% (MOD) of maximal oxygen consumption. Ambulating BP was measured for 14 h after each experiment. RAAS polymorphisms associated with hypertension (i.e. angiotensin converting I enzyme, ACE I/D; angiotensin II type 1 receptor, AT1R A/C; and intron 2 of aldosterone synthase, Int2 W/C) were analyzed using polymerase chain reaction and restriction enzyme digestion. Repeated measure ANOVA tested if BP differed between experimental conditions by RAAS genotypes. Compared to men with 0-2 variant alleles, men with > or =3 combined RAAS variant alleles had lower average systolic BP (SBP) (P=0.030) and lower average diastolic BP (DBP) (P=0.009) for 14 h only after LITE. In contrast, average BP was not different for MOD and control between RAAS variant allele groups over this time period (P> or =0.05). LITE reduced BP in men with > or =3 variant RAAS alleles for 14 h, whereas MOD had no influence on BP in these men. In order to optimally prescribe exercise for its BP lowering benefits in those with hypertension, additional knowledge of how genetic variation affects the BP response to exercise is needed. PMID- 16468059 TI - Vmax estimate from three-parameter critical velocity models: validity and impact on 800 m running performance prediction. AB - The purpose of this study was to evaluate the validity of maximal velocity (Vmax) estimated from three-parameter systems models, and to compare the predictive value of two- and three-parameter models for the 800 m. Seventeen trained male subjects (VO2max=66.54+/-7.29 ml min(-1) kg(-1)) performed five randomly ordered constant velocity tests (CVT), a maximal velocity test (mean velocity over the last 10 m portion of a 40 m sprint) and a 800 m time trial (V 800 m). Five systems models (two three-parameter and three two-parameter) were used to compute V max (three-parameter models), critical velocity (CV), anaerobic running capacity (ARC) and V800m from times to exhaustion during CVT. Vmax estimates were significantly lower than (0.1910) and healthy volunteers. INTERVENTIONS: Fatigue was evoked during 20[Symbol: see text]min of intermittent tetanic ulnar nerve stimulation achieving 50% of maximum AP muscle force. MEASUREMENTS AND RESULTS: We measured evoked AP muscle force and fatigue, and compound muscle action potential (CMAP), and performed standard electrophysiological tests in 13 patients, and in 7 volunteers before and after immobilization. Maximum force (20+/-16 vs 65+/-19N; p<0.01) and CMAP (3.6+/-2.5 vs 10+/-2.5 mV; p<0.05) were markedly decreased in patients; however, fatigue and ulnar nerve conduction velocity did not differ from volunteers, and a decrement of CMAP was not observed with nerve stimulation frequencies up to 40 Hz. All patients with critical illness polyneuropathy, and an additional 50% of those without, had significant muscle weakness. CONCLUSION: Peripheral muscle force is markedly decreased in sepsis, without evidence for an increased fatigability. Muscle weakness was most likely due to a sepsis-induced myopathy and/or axonal neuropathy, and was not the result of an immobilization atrophy. PMID- 16468075 TI - JWA, a novel signaling molecule, involved in all-trans retinoic acid induced differentiation of HL-60 cells. AB - JWA (AF070523) was originally identified as a novel all-trans retinoic acid (ATRA) responsible gene in primary human tracheal bronchial epithelial cells. For the notable performance achieved by ATRA in the differentiation induction therapy, we investigated the role of JWA in ATRA-mediated differentiation of the human myeloid leukemia HL-60 cells. We found that concomitant with the progressive cell differentiation, JWA expression was up-regulated by ATRA in a dose- and time-dependent manner. Inhibition of JWA expression by RNA interference partially blocked ATRA-induced differentiation and growth inhibition of HL-60 cells. Pre-treatment of phorbol-12-myristate-13-acetate (TPA), a PKC activator, decreased ATRA-mediated differentiation, companied with the down-regulation of JWA expression. Arsenic trioxide (As(2)O(3), 0.5 microM) enhanced the cellular differentiation induced by 0.01 microM ATRA, but had no noticeable effect on the differentiation induced by 0.1 microM ATRA. Concurrent with the enhancement, JWA expression was up-regulated. All the data suggest that up-regulation of JWA expression is essential for ATRA-induced differentiation of HL-60 cells. And JWA, associated with PKC, is involved in its signal pathways. Ideal therapeutic efficacy with low toxicity may be obtained if low doses of ATRA (0.01 microM) and As(2)O(3) (0.5 microM) are combined. These findings may present a novel mechanism that cellular differentiation and growth inhibition induced by ATRA are mediated at least in part through regulation of JWA expression. JWA may be a novel molecular marker for ATRA-induced HL-60 cell differentiation. ATRA up-regulates JWA expression by stimulating the transcriptional activity of JWA gene promoter. PMID- 16468076 TI - Assessing measurement properties of two single-item general health measures. AB - BACKGROUND: Multi-item health status measures can be lengthy, expensive, and burdensome to collect. Single-item measures may be an alternative. We compared measurement properties of two single-item, general self-rated health (GSRH) questions to assess how well they captured information in a validated, multi-item instrument. METHODS: We administered a general health survey (SF-12V) that included "standard" and "comparative" forms of a GSRH. We repeated the survey two weeks later to the same 75 medically stable outpatients to test for GSRH reproducibility, reliability, and validity using SF-12V Physical Functioning and Emotional Health subscales as a reference. RESULTS: At each survey administration, the two GSRH questions demonstrated good alternate forms reliability (first administration: r = 0.74, p < 0.001; second administration: r = 0.74, p < 0.001) and good reproducibility ("standard": ICC 0.69; "comparative": ICC 0.85). Both GSRH items correlated with physical functioning ("standard": r = 0.66; "comparative": r = 0.56) and emotional health measures ("standard": r = 0.65; "comparative": r = 0.59). Mean subscale measures associated with responses in each GSRH category were significantly different (ANOVA, p < 0.001), indicating strong discriminant scale performance. CONCLUSIONS: Our single-item, GSRH questions demonstrated good reproducibility, reliability, and strong concurrent and discriminant scale performance with an established health status measure. PMID- 16468077 TI - The PedsQL 4.0 as a school population health measure: feasibility, reliability, and validity. AB - BACKGROUND: The application of health-related quality of life (HRQOL) as a school population health measure may facilitate risk assessment and resource allocation, the tracking of student health at the school and district level, the identification of health disparities among schoolchildren, and the determination of health outcomes from interventions and policy decisions at the school, district, and county level. OBJECTIVE: To determine the feasibility, reliability, and validity of the 23-item PedsQL 4.0 (Pediatric Quality of Life Inventory) Generic Core Scales as a school population health measure for children and adolescents. DESIGN: Survey conducted in 304 classes at 18 elementary schools, 4 middle schools, and 3 high schools within a large metropolitan school district. METHODS: The PedsQL 4.0 Generic Core Scales (Physical, Emotional, Social, School Functioning) were completed by 2437 children ages 8-18 and 4227 parents of children ages 5-18. RESULTS: The PedsQL 4.0 evidenced minimal missing responses, achieved excellent reliability for the Total Scale Score (alpha = 0.89 child, 0.92, parent report), and distinguished between healthy children and children with chronic health conditions. The PedsQL 4.0 was related to indicators of socioeconomic status (SES) at the school and district level. The PedsQL School Functioning Scale was significantly correlated with standardized achievement scores based on the Stanford 9. CONCLUSION: The results demonstrate the feasibility, reliability and validity of the PedsQL 4.0 Generic Core Scales as a school population health measure. The implications of measuring HRQOL in schoolchildren at the school, district, and county level for identifying and ameliorating health disparities are discussed. PMID- 16468078 TI - The validity and reliability of the functional impairment checklist (FIC) in the evaluation of functional consequences of severe acute respiratory distress syndrome (SARS). AB - Severe acute respiratory distress syndrome (SARS) contributed to significant mortality and morbidity worldwide. We aimed to establish the validity, reliability and responsiveness of the functional impairment checklist (FIC) as a measurement tool for physical dysfunction in SARS survivors. One hundred and sixteeen (65 females and 51 males, mean age 45.6) patients who joined the SARS rehabilitation programme were analysed. The factor analysis yielded two latent factors. The mean FIC-symptom and FIC-disability score were 24.12 (SD +/- 20.2) and 26.11 (SD +/- 27.32), respectively. Based on the item-scale correlation coefficients, the Cronbach's alpha coefficients reflecting the internal consistency reliability of scale score were 0.75 for FIC-symptom and 0.86 for FIC disability. Test-retest reliability in 23 patients showed no statistical significant difference in the FIC scores between tests with intraclass correlation coefficient (ICC) 0.49-0.57. The FIC scales correlated both with 6 munute walking test (6MWT) distance (-0.26 and -0.38) and handgrip strength (HGS) (-0.20 and -0.27). Moreover, the FIC scales correlated with St. George's respiratory questionnaire (SGRQ) (0.19 to 0.52) and short form 36 Hong Kong (SF 36) domains (-0.19 to -0.59). Both FIC scales correlated stronger with physical component summary (PCS) (-0.41 and -0.55) than with mental component summary (MCS) (-0.30 and -0.23). FIC reduced significantly at 6 months while the SF-36 PCS and MCS did not show any change. In conclusion, the study results indicate the FIC is reliable, valid and responsive to change in symptom and disability as a consequence of SARS, suggesting it may provide a means of assessing health related quality of life (HRQOL) outcomes in a longitudinal follow up. PMID- 16468079 TI - Evaluation of cross-sectional and longitudinal construct validity of two vision related quality of life questionnaires: the LVQOL and VCM1. AB - The Low Vision Quality of Life (LVQOL) questionnaire and the Vision-related Quality of Life Core Measure (VCM1) are two of the many vision-related quality of life (QOL) questionnaires that have been developed in recent years. Although psychometric properties of the LVQOL and VCM1 compare well with other vision related QOL questionnaires, construct and longitudinal validity have not been assessed (adequately). The purpose of this study was to examine the cross sectional and longitudinal construct validity of these questionnaires by testing specific pre-specified hypotheses about the relations of these questionnaires with other measures. The percentage of hypotheses regarding the cross-sectional construct validity that were refuted for the LVQOL was 22% for the basic aspects of vision subscale, 50% for the mobility subscale, 39% for the adjustment subscale and 17% for the reading and fine work subscale. For the VCM1 this percentage was 57%. For the longitudinal construct validity the percentage of hypotheses that were refuted ranged from 33 to 75% for the LVQOL subscales and was 50% for the VCM1. In conclusion, cross-sectional construct validity was satisfactory for the LVQOL subscales, but seemed poor for the VCM1. In addition, the longitudinal validity of these scales was poor to moderate. PMID- 16468080 TI - Changes in health status, psychological distress, and quality of life in COPD patients after hospitalization. AB - The purpose of this paper was to describe quality of life (QoL) following an acute exacerbation of chronic obstructive pulmonary disease (COPD), and to examine possible relationships between QoL, health status, psychological distress and QoL. This prospective longitudinal study examined data from hospitalization and two subsequent phases, collected over a 9-month period. The sample consisted of 51 COPD patients aged 48-87 years. Health status was assessed with the St George's Respiratory Questionnaire (SGRQ); psychological distress with the Hopkins Symptom Checklist (HSCL); and quality of life using the World Health Organization Quality of Life - Bref (WHOQOL - Bref). Health status improved significantly over the 9 months; from 65.95 to 59.40 (p = 0.001) in the SGRQ total score. Psychological distress improved significantly from hospitalization to the 1-month assessment (T2) (p = 0.001). QoL remained stable except for a significant increase in the physical domain from hospitalization to T2, and in the environmental domain from hospitalization to T2 and T3. Test-retest correlations of the WHOQOL - Bref were high. The results suggest an improvement in the health status over a 9-month period from being discharged after an exacerbation of COPD in spite of high levels of psychological distress and reduced QoL. PMID- 16468082 TI - Stated preferences of patients with cancer for health-related quality-of-life (HRQOL) domains during treatment. AB - OBJECTIVES: It is postulated that patients with different cancer diagnoses, stages of disease and treatments will exhibit different individual preferences for health-related quality-of-life (HRQOL) functional domains and symptoms. METHODS: A stated-preference (SP) instrument incorporating all functional domains and symptoms of the EORTC Quality of Life Questionnaire (QLQ-C30) was administered to 400 patients with either breast (n = 150); colorectal (n = 150) or non-small cell lung cancer (n = 100) who had previously experienced chemotherapy. The SP survey asked patients to make choices between a series of hypothetical functional/symptom pairs defined by combinations of HRQOL attributes, and depicted by levels of functioning and symptomatology. RESULTS: In the 400 patients, considered as one group, role, cognitive, and social functioning, fatigue, nausea/vomiting, pain, appetite loss, diarrhea and financial difficulties were most important, whereas physical and emotional functioning, dyspnea, constipation and insomnia were less important. The four effects that patients with breast cancer most wished to avoid were nausea and vomiting, pain, and decreases in emotional and role functioning. Patients with colorectal cancer listed diarrhea as the second most important effect to avoid (after nausea/vomiting, but before pain and role functioning), whereas those with non-small cell lung cancer listed dyspnea as the fourth most important effect to avoid. CONCLUSION: These results provide more precise information regarding patient treatment concerns than that provided by the usual measurement of HRQOL. This information can be used by clinical trial investigators to design more precise interventions to improve HRQOL in the domains of greatest importance to patients and by all health care professionals to improve counseling of patients. PMID- 16468081 TI - Health-related quality of life in multiple sclerosis: The impact of disability, gender and employment status. AB - OBJECTIVES: (1) Evaluate the impact of the patient characteristics of disability, gender and employment status on health-related quality of life (HRQOL) in multiple sclerosis (MS) and (2) Characterize the functional relationship between HRQOL and disability overall, and by gender and employment status. METHODS: We assessed the HRQOL of 215 MS outpatients in our clinic using the MSQOL-54 and Fatigue Severity Scale (FSS), and that of 172 healthy controls, using the SF-36 (a subset of MSQOL-54). We compared QOL between MS subgroups defined by disability, gender and employment, and computed the linear and non-linear relationships between disability level measured by the Expanded Disability Status Scale (EDSS) and MSQOL-54 dimensions. RESULTS: QOL of MS patients measured by SF 36 is lower than controls, varying by QOL dimension with the greatest difference emerging for physical aspects of the disease. The relationship of physical disability, measured by EDSS, and all 14 MSQOL-54 dimensions and FSS is negative; for 12 of the 14 dimensions and FSS it is also non-linear. Non-linearity is most pronounced among women, who show a weak EDSS/QOL relationship at higher levels of physical disability, suggesting women better able to "psychologically buffer" the debilitating aspects of MS. While employed have higher QOL than unemployed, the former are more affected by physical disability. CONCLUSIONS: Multiple attributes, including disability, gender and employment status, affect QOL. QOL's relationship with disability is complex, displaying non-linearity and interacting with patient characteristics. This has implication for QOL research methodology and provides insight into factors affecting patients' perceptions of well-being. PMID- 16468083 TI - Are chemotherapy patients' HRQoL importance weights consistent with linear scoring rules? A stated-choice approach. AB - OBJECTIVE: To compare a linear scoring rule with the subjective importance of different domain and symptom levels of the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-C30 (EORTC QLQ-C30) among patients undergoing chemotherapy. METHODS: Using a stated-choice or choice-format conjoint analysis survey instrument, we elicited patient preferences for varying levels of physical, role, social, emotional, and cognitive function along with chemotherapy-related side effects and financial difficulties. A total of 375 patients completed the questionnaire: 159 breast cancer, 117 colorectal cancer, 99 non-small-cell lung cancer; and 21 with unknown tumor type. Constrained maximum likelihood estimates were used to estimate relative importance weights for each level of each domain and symptom. RESULTS: Summary HRQoL measures generally presume that differences among Likert categories are equally important to patients within and across outcomes. Our results indicate strong non linearities both within and across domain and symptom categories. Improvements from severe pain to mild pain, severe fatigue to no fatigue, and severe social limitations to moderate social limitations are all about twice as important as no work to limited work in the Role domain. CONCLUSIONS: Our results indicate large differences in the impact of individual domains and symptoms on patient perceptions of well-being. Most cancer patients are likely to be less concerned about specific symptoms than the impact of those symptoms on their ability to function physically, socially, and in their daily roles. PMID- 16468084 TI - Responsiveness of the National Institutes of Health Chronic Prostatitis Symptom Index (NIH-CPSI). AB - OBJECTIVES: The NIH-Chronic Prostatitis Symptom Index (NIH-CPSI) was developed to assess symptoms and quality of life in men with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS). We assessed the responsiveness of the NIH-CPSI to change over time and defined thresholds for changes perceptible to patients. METHODS: We studied 174 men with CP/CPPS who participated in a placebo-controlled randomized clinical trial. Changes from baseline to six weeks in the NIH-CPSI total score and pain, urinary, and quality of life subscores were compared to a global response assessment (GRA). Effect sizes and Guyatt statistics were calculated to evaluate responsiveness; 95% confidence intervals were produced using bootstrapping. RESULTS: All scores decreased over time with the largest decrease in subjects who reported on the GRA that they were markedly improved. The NIH-CPSI total, pain, and quality of life scores were highly responsive in the improved groups; the urinary score showed minimal responsiveness. There was no evidence of responsiveness among those subjects who worsened on the trial. ROC curves identified a 6-point decline in the NIH-CPSI total score as the optimal threshold to predict treatment response. CONCLUSIONS: The NIH-CPSI total score and pain and quality of life subscores are responsive to change over time. PMID- 16468085 TI - A brief mental health outcomes measure: translation and validation of the Czech version of the Schwartz Outcomes Scale-10. AB - The Schwartz Outcomes Scale-10 (SOS-10) is a 10-item scale developed to measure the effectiveness of psychiatric treatments. Using standard methodology, we translated the scale into Czech and examined the psychometric properties of the Czech version. 207 in-patients admitted to Prague Psychiatric Center were included in the study. All patients completed the SOS at admission and discharge. The SOS-10 scale was also administered to 170 persons from the general population. Reliability, validity and sensitivity to treatment change of the Czech SOS-10 were analyzed. The Cronbach's alpha coefficient was 0.92. The item total correlation coefficients varied from 0.56 to 0.82. The SOS-10 correlated well with condition-specific measures including depression (BDI) and anxiety (BAI) and a global self-rated symptom severity scale (CGI). The SOS-10 also had significant correlations with the Health, Basic needs, Relationship, and Leisure time domains of the Czech version of the Subjective Quality of Life Analysis (SQUALA-CZ). The scale discriminated well between patients and controls, with patients scoring significantly lower on all SOS items. The patient sample's admission and discharge scores were significantly different, indicating that the scale is sensitive to treatment changes. We concluded that the Czech SOS-10 is valid with reliability and factor structure similar to the American language version. PMID- 16468087 TI - Knowledge and attitudes toward a free education and Ashkenazi Jewish carrier testing program. AB - Carrier testing is offered on the basis of Ashkenazi Jewish background in both the prenatal and preconception settings, with the goal of decreasing the prevalence of affected individuals and allowing informed decision-making during childbearing. The purpose of this study was to (1) document the demographic characteristics of individuals who attended a free education and screening program, (2) learn how the education program changed attendees' knowledge and attitudes by learning more about these disorders, and (3) determine how participants perceived their carrier status risk. One hundred seventy-four individuals completed questionnaires at the beginning and end of an educational program about the Ashkenazi Jewish genetic disorders. There was a statistically significant difference in the participant's level of knowledge from the pre- to post education (p < .001). Females reported a significantly higher level of concern about the disorders (p = .004) and their carrier status (p = .006) before the education, as well as about their carrier status post education (p = .05). Finally, having one or more parent affiliated with Orthodox Judaism was related to higher knowledge before the education program (p = .05). In conclusion, this study demonstrated that an educational carrier screening program increased knowledge about the disorders and also produced mild anxiety regarding personal and reproductive risks. PMID- 16468088 TI - The importance of written information packages in support of case-finding within families at risk for inherited high cholesterol. AB - Inherited High Cholesterol is treatable, but highly underdiagnosed. To detect undiagnosed blood relatives at a presymptomatic stage, in the Netherlands written information packages are available to facilitate family communication. To investigate the role of those packages in the detection of carriers, we conducted a qualitative evaluation (plus-minus method combined with semistructured interviews with index patients and relatives). Our data suggest that interviewees approved the family approach for finding carriers, although reluctantly. The packages aided family disclosure by reducing hesitation. However, index patients only informed first-degree relatives and generally communicated the risk only once. This may be due to the cultural context and a limited understanding of genetics. For relatives the packages served as a cue to action and as a legitimation to gain access to a diagnostic cholesterol test. Despite the value of these written materials, they should not be used as the only communication between index patient and relatives. PMID- 16468089 TI - The role of maternal acceptance in the relation between community violence exposure and child functioning. AB - Children in the United States are exposed to considerable community violence that has been linked to child functioning. However, not all those exposed, experience negative outcomes. Recent research has focused on factors that "buffer" or protect children from negative consequences of violence exposure. The purpose of this investigation was to examine the potential buffering or moderating role of maternal acceptance in the relationship between community violence exposure and internalizing and externalizing problems. Subjects were 268 urban African American first graders. Community violence exposure was significantly related to symptoms of post-traumatic stress, but did not correlate with either internalizing or externalizing problems for all children, after control for demographics, maternal mental health, and general life stress. However, children's perceptions of maternal acceptance moderated the relationship between violence exposure and internalizing and externalizing problems which included being withdrawn, anxious-depressed, and demonstrating delinquent behavior. Children with the lowest levels of self-reported maternal acceptance were most impacted by community violence. In this sample of urban first graders, low levels of maternal acceptance placed children at greater risk for adverse outcomes associated with community violence exposure compared to moderate and high levels of maternal acceptance. PMID- 16468091 TI - A modeling study of notch noise responses of type III units in the gerbil dorsal cochlear nucleus. AB - A computational model of the neural circuitry of the gerbil dorsal cochlear nucleus (DCN), based on the MacGregor's neuromime model, was used to simulate type III unit (P-cell) responses to notch noise stimuli. The DCN patch model is based on a previous computational model of the cat DCN [Hancock, K. E., and H. F. Voigt. Ann. Biomed. Eng. 27:73-87, 1999]. According to the experimental study of Parsons et al. [Ann. Biomed. Eng. 29:887-896, 2001], the responses of gerbil DCN type III units to notch noise stimuli are similar to those of cat DCN type IV units, which are thought to be spectral notch detectors. This suggests that type III units in the gerbil DCN may serve as spectral notch detectors. In this modeling study, a simplified notch noise response plot-spike discharge rate vs. notch cutoff frequency plot-was used to compare model responses to the experimental results. Parameter estimation and sensitivity analysis of three connection parameters within the DCN patch have been studied and shows the model is robust, providing reasonable fits to the experimental data from 14 of 15 type III units examined [work supported by a grant from NIDCD, Boston University's Biomedical Engineering department and Hearing Research Center]. PMID- 16468090 TI - Optimization of a dilution method for human expanded blastocysts vitrified using EM grids after artificial shrinkage. AB - PURPOSE: To verify a more effective dilution method that can be applied to human expanded blastocysts that are vitrified after artificial shrinkage. METHODS: Surplus expanded blastocysts that remained after embryo transfer (ET) in in vitro fertilization (IVF) cycles, were cryopreserved. The blastocysts were vitrified on EM grids following artificial shrinkage. After thawing the blastocysts, cryoprotectants were diluted using either a 6- or 2-step method. We examined the survival rate and clinical outcome of blastocysts of 151 patients in our ET program after thawing. RESULTS: The survival rate of blastocysts that were thawed using a 2-step method (91.6%, 239/261) was comparable with that of the 6-step method (89%, 186/209). The clinical pregnancy rate (45.9%, 39/85) and implantation rate (24.1%, 53/220) were slightly higher in the 2-step method than in the 6-step method (40.9%, 27/66; 19.4%, 33/170). CONCLUSIONS: Our data indicate that the 2-step dilution method could be a simpler and more effective protocol for human expanded blastocysts that are vitrified using EM-grid following artificial shrinkage. PMID- 16468092 TI - Influence of multiple stenoses on echo-Doppler functional diagnosis of peripheral arterial disease: a numerical and experimental study. AB - The objective of this paper was to evaluate the ability of the peak systolic velocity ratio (PSVR) and pressure drop (DeltaP) to detect and grade multiple stenoses in lower limb mimicking arteries. Numerical simulations and experiments in vascular phantoms allowing ultrasound duplex scanning and pressure measurements were used to investigate simple and double stenotic arterial segments. Inter-stenotic distance, severity of the distal stenosis, flow rate and flow profile (steady or pulsatile) were the tested parameters. The three dimensional simulations considered the turbulent two-equation Wilcox model. Agreements were observed between the experimental and numerical results for DeltaP and PSVR. The maximum PSVR along the artery was shown to be mainly influenced by the severity of the most important stenosis. However, mutual interactions of both stenoses on hemodynamics were noted. By using the clinical PSVR threshold used to diagnose critical lesions (PSVR > or = 2), its longitudinal evolution along the artery poorly reflected the length of the lesion or the impact of surrounding stenoses. This investigation confirms the interaction between adjacent stenoses on hemodynamics and its impact on the Doppler ultrasound index PSVR. PMID- 16468097 TI - [Principles and perspectives -- advances and future of medicine]. PMID- 16468093 TI - The estimation of lung mechanics parameters in the presence of pathology: a theoretical analysis. AB - Mechanical lung function is frequently assessed in terms of lung resistance (R (L)), lung elastance (E (L)), and airway resistance (R (aw)). These quantities are determined by measuring input impedance at various oscillation frequencies, and allow lung tissue resistance (R (t)) to be estimated as the difference between R (L) and R (aw). These various parameters change in characteristic ways in the presence of lung pathology. In particular, the ratio R (t)/E (L) (known as hysteresivity, (eta) has been shown both experimentally and in numerical simulations to increase when regional heterogeneities in mechanical function develop throughout the lung. In this study, we performed an analytical investigation of a two-compartment lung model and showed that while heterogeneity always leads to an increase in E (L), eta will increase only initially. When heterogeneity becomes extreme, eta stops increasing and starts to decrease. However, there are no experimental reports of eta decreasing under conditions in which heterogeneity would be expected to exist. We speculate that this is because liquid bridges invariably form across airway lumen that narrow to a certain point, thereby preventing them from achieving arbitrarily small non-zero radii. We also show that recruitment of closed lung units during lung inflation may lead to variables responses in both eta and E (L). PMID- 16468098 TI - [Fast track to a university chair -- with the support of Emmy Noether]. PMID- 16468099 TI - [Pharmacogenomics: principles and perspectives]. PMID- 16468100 TI - [Changes in the perception of risk factors for stroke in the German population between 1995-1996 and 2000-2001. Results of two representative EMNID surveys]. AB - BACKGROUND AND OBJECTIVE: Stroke is one of the leading causes for death and disability worldwide. A better understanding of the perception of modifiable stroke risk factors in the population is the first step to initiate effective prevention strategies on population level. Changes over 5 years in the risk perception in the general population were investigated by two representative surveys in Germany. METHODS: Nationwide programs to screen voluntary participants for stroke risk were undertaken in 1995-1996 and 2000-2001 by the German Stroke Foundation, in cooperation with the health insurance company BARMER and the Sanofi-Synthelabo Company. As part of these programmes two surveys were performed by TNS-EMNID to collect data on population knowledge. A representative sample of the German population was selected and asked to categorize their perception of stroke risk for common vascular risk factors. RESULTS: A total of 8193 participants were interviewed (4081 in 1995-1996 and 4112 in 2000-2001); 43.5% were (3) 50 years of age and 52.5% were female. Hypertension was rated by 68.3% to be in the highest risk category for stroke, followed by smoking (52.3%), hypercholesterolemia (48.0%), overweight (48.0%), excessive alcohol consumption (32.9%) and diabetes (26.6%). The proportion of participants who graded these factors to be important for stroke occurrence was persistently higher in 2000 2001 than in 1995-1996. CONCLUSION: Perception of modifiable risk factors for stroke increased over a 5-year time period in two representative surveys in Germany. The importance of diabetes mellitus as a risk factor for stroke is especially underestimated in the general population. PMID- 16468101 TI - [Fire-eater's lung. Two cases and review of the literature]. AB - HISTORY AND ADMISSION FINDINGS: Two men were hospitalized (three years apart) after aspirating petroleum on their fist attempt at "fire-eating". Pt.1 (aged 25 years) complained of chest pain, dyspnea and dizziness. The other (Pt. 2; aged 29 years) had a hemoptysis. Pt. 1 had a normal body temperature, but the breath sounds were decreased over the left base. The breath sounds in Pt. 2 were normal, but he had a fever of 38.8 degrees C. Routine physical examination was unremarkable. INVESTIGATIONS: Laboratory tests in both patients revealed increased inflammatory parameters. Chest radiographs showed that Pt. 1 had a left basal alveolar infiltrate, while Pt. 2 had an infiltrate in the right middle and upper lobe, which on computed tomography after a few days showed signs of cavitation in the left infrahilar region and the middle lobe, respectively. Pneumococci were found in the sputum of Pt. 2. TREATMENT AND COURSE: Both patients were given antibiotics, Pt. 1 also had salbutamol inhalation treatment. The patients were discharged symptom-free after 8 and 10 days, respectively. A follow-up chest radiograph four weeks later in Pt. 1 merely showed streaky fibrotic residues. CONCLUSION: These two case reports and detailed literature search indicate that immediate administration of antibiotics is important, while routine steroid treatment is not necessary. PMID- 16468103 TI - [Immunogenic myositis]. AB - Immunogenic inflammatory myopathies are a heterogeneous group of acquired muscle disorders. Clinical and morphological characteristics are on one side muscle weakness, on the other side inflammatory infiltrates in muscle biopsy. Three main groups of different pathogenesis and course can be subdivided: The treatment of inflammatory myopathies is predominantly based on empiric data. Baseline drugs are Corticosteroids and Immunosuppressives. High dose intravenous Immunoglobulins (IVIG) are an important additional therapeutic possibility, especially in inclusion body myositis. PMID- 16468106 TI - [Current position regarding the diagnosis of tuberculosis]. PMID- 16468107 TI - [Hyponatremia with clinical relevance]. PMID- 16468108 TI - [Rehabilitation of children and adolescents]. AB - In recent years a significant increase in most chronic mental and physical illnesses among children and adolescents was reported. Thus, medical child and adolescent rehabilitation plays a major role. Therefore, early diagnosis and multimodal intervention in childhood and adolescence is necessary, to prevent a chronic course of disease. Patient education programs have been shown to be effective in the treatment of chronic illnesses. Those programs are mostly aimed to improve the compliant behavior and disease management. Consecutively, the health status and quality of life should be improved. Due to the implementation of guidelines and evaluation studies the quality of rehabilitation programs was ameliorated during the past 15 years. However, future research should develop flexible rehabilitation programs in particular (rehabilitation duration, rehabilitation near or far from place of residence, rehabilitation chain with in patient, partially out-patient, and out-patient programs), whose effectiveness has to be proven. PMID- 16468109 TI - [Structure quality assurance in in-patient medical rehabilitation of children and adolescents]. AB - BACKGROUND: So far, the routine programmes of the statutory pension and health insurance institutes for quality assurance in medical rehabilitation have not included facilities for children and adolescents. Thus, a conjoint project of the statutory pension and health insurance institutes to assess and to describe the structure quality of in-patient medical rehabilitation clinics for children and adolescents was initiated. The aim of the project was a survey and an analysis of the currently existing clinic structures and the development of criteria for an evaluation of the structure quality. The Institute of Medical Psychology, University of Hamburg and the Department of Quality Management and Social Medicine, University of Freiburg were engaged to conduct the project. METHOD: A questionnaire was developed in a multi-stage process with expert involvement which covers general information, characteristics of structure quality as well as structure affiliated process characteristics of clinics. The survey was planned as a cross-sectional study. Data collection was accomplished from 1.12.2004 - 15.3.2005. RESULTS: A total of n = 177 facility addresses were available. The return rate was 83.3 %. Eventually, n = 79 facilities were included in the analysis. In sum, the results underline the heterogeneity of services in the field of child and adolescent rehabilitation. CONCLUSIONS: The results allow an analysis of the current clinic structures as well as a first comparison of clinics on the basis of the assessed structure characteristics. PMID- 16468110 TI - [Medication behavior of children and adolescents with asthma before and after in patient rehabilitation--results of a multicenter study]. AB - The article presented is based on data of a multicenter study examining the effectiveness of an inpatient rehabilitation program for children and adolescents with asthma. The study took place from August 1996 to November 2001. It shows more noticeable long-term improvement of disease management and lung function parameters after in-patient rehabilitation (n = 226) than after out-patient rehabilitation programs (n = 92). The re-analysis of the data examines the influence of medication on these results. A decisive effect on treatment results for both in- and outpatients can be achieved by complying with the guidelines recommended by the medical societies. However, at the same time the results demonstrate that prescribing a correct treatment plan is not always sufficient. The significant improvement of outcome parameters (functional restriction, lung function, days absent from school, visits to a physician) of the rehabilitation group shows the importance of rehabilitation for teaching strategies for disease management and increase in compliance. PMID- 16468111 TI - [Education of children with epilepsy and their parents by the modular education program epilepsy for families (FAMOSES)--results of an evaluation study]. AB - PURPOSE: The aim of the study was to evaluate the efficacy of the modular educational program for children with epilepsy and their parents (FAMOSES). This program was developed by an interdisciplinary project group to improve knowledge, coping, treatment outcome, emotional and practical adaptation to the condition. METHODS: A prospective, controlled, multi-center, pre-post study design was used to examine the efficacy of the program in the treatment group compared to the waiting group (control group). Questionnaires included epilepsy specific scales regarding knowledge, attitudes, restrictions in daily living, epilepsy related fears, coping with the chronic disease and generic instruments (quality of life, KINDL). 55 parents of the treatment group completed the questionnaires three months before the course and three months later; the corresponding waiting group included 48 parents. Respectively, 31 children, who participated in the program, completed the questionnaires immediately before the course and three months later; the corresponding waiting group included 19 children. RESULTS: Children, who attended the program, showed improvements in the domains perceived restrictions (significant, medium effect size), absence from school and seizure frequency. Not significantly greater compared to the control group were the improvements of knowledge, attitudes and fears regarding to the epilepsy. Parents of the treatment group showed significant enhancements in epilepsy specific knowledge (large effect size), attitudes toward the epilepsy, management of epileptic seizures and significant reductions of fears and restrictions of their child with epilepsy (small to medium effect sizes). PMID- 16468112 TI - [Effects of a programme for structured outpatient follow-up care after inpatient rehabilitation of obese children and adolescents--a multicentre, randomized study]. AB - INTRODUCTION: Inpatient rehabilitation is an important contributor to the treatment of child and adolescent obesity with comorbidity in Germany. Thus far outpatient follow-up care has not been subject to research. The main goal of this study is the development and longer-term evaluation of a programme for structured outpatient follow-up care by primary care physicians after inpatient rehabilitation of obese children and adolescents. METHODS: This multicentre, randomized, parallel controlled clinical trial enrolled 521 children and adolescents aged 9 - 16 (avg. 13.3) years in 7 cooperating specialized rehabilitation facilities. The patients were randomized at the end of their multimodal inpatient treatment that lasted for 5.9 (+/- 1.2) weeks. The intervention consisted of 12 half-hour outpatient consultations by the primary care physicians utilizing a modular consultation guideline and an internet-based training programme. The control group received standard care. Outcome variables included anthropometric measures, questionnaires on eating behaviour, physical activity, quality of life, self-efficacy and subjective rating of the intervention. Measurements were performed at the beginning (t1) and end (t2) of inpatient rehabilitation as well as 6 (t3) and 12 (t4) months after discharge. RESULTS: There was a highly significant improvement of BMI-z-score (standard deviation score of the Body Mass Index), relevant behaviour, quality of life and self-efficacy between t1 and t4. On intention to treat analysis no differences were detected between intervention and control groups. The intervention was utilized by only 50 % of entitled. The BMI-z-score reached a minimum at t3 (6 month follow-up). Predictors of longer-term BMI-z-score decline were psychological suffering, poor eating behaviour, low physical activity and high socio-economic status. In multivariate linear regression improvement of eating behaviour and physical activity correlated with the improvement of BMI-z-score. DISCUSSION: This low-level outpatient intervention did not improve the demonstrated longer-term effectiveness of inpatient rehabilitation. An interdisciplinary outpatient follow-up care in groups with inclusion of the parents is recommended. PMID- 16468113 TI - [Multimodal therapy of children with attention deficit/hyperactivity disorder and their parents in in-patient rehabilitation]. AB - BACKGROUND: Attention deficit/hyperactivity disorder (ADHD) is one of the most frequent psychic disorders of childhood, characterized by high persistence and an increased risk for the development of externalising disorders of behavior. Due to ADHD increased problems can be frequently observed at school as well as in the social and emotional context of the child. Concepts for in-patient rehabilitation have not been developed, although effective modifications of the behavior of ADHD patients appeared to be achievable under the conditions found in in-patient rehabilitation. OBJECTIVE: The present pilot study aimed to evaluate whether this newly developed concept has any benefit on psychological problems, coping and quality of life. METHOD: In total, n = 28 boys with ADHD 7 to 12 years old were included into a quasi-experimental pre-post design and completed self-report questionnaires on psychological problems, coping and quality of life. In addition, 26 mothers evaluated the behavior problems, psychological problems and quality of life of their sons at the beginning and the end of in-patient rehabilitation. During a 4-week in-patient rehabilitation all children and their mothers took part in a multimodal training containing components of behavior therapy and cognitive-behavioral therapy (self-instruction, self-management, and stress management), applied in child-, parent-child- and parent-centered interventions. RESULTS: Parents estimated the behavior problems of their children as significantly reduced after in-patient rehabilitation as compared to before. In addition, psychological problems and quality of life improved significantly in self- and parent-proxy reports during the in-patient rehabilitation. CONCLUSION: Our results support the hypothesis that this multimodal therapy concept is an effective tool for the treatment of ADHD patients. However, a replication of our results in a control-group study on a larger population is required. PMID- 16468114 TI - [Report of the annual convention of the Lower Saxony/Bremen rehabilitation research network on theme "From Health Economics to Rehab Economics", November 11, 2005 in Braunschweig]. PMID- 16468115 TI - Extreme hyperkalemia. PMID- 16468116 TI - US Senators introduce bill to accelerate AIDS vaccine research. PMID- 16468117 TI - Stress management. PMID- 16468118 TI - Grooming women entrepreneurs. PMID- 16468119 TI - [Ocular allergies]. AB - Recent developments indicate that ocular allergy is more than an IgE-mediated allergic conjunctivitis. Ocular allergy is a disease affecting the entire ocular surface including conjunctiva, lids, cornea, lacrimal gland and tear film. Besides an IgE-mediated reaction, a complex chronic inflammation is involved in the pathogenesis of many ocular allergies. According to their pathogenesis and clinical picture, ocular allergies are classified into mild forms, such as seasonal and perennial allergic conjunctivitis as well as giant papillary conjunctivitis, and chronic, potentially blinding forms such as atopic keratoconjunctivitis and vernal keratoconjunctivitis. New therapeutics act on the entire inflammatory process or try to modulate the allergic reaction early and specifically. The association with non-ocular allergic symptoms requires an interdisciplinary approach. PMID- 16468120 TI - [Abecedary of the "Annales". Part 9]. AB - The terms included and detailed in the present part are: Standard error, Clinical trial, Randomized controlled trial, Cross-over clinical trial, Equivalence trial and non inferiority trial. PMID- 16468121 TI - A road map for efficient and reliable human genome epidemiology. AB - Networks of investigators have begun sharing best practices, tools and methods for analysis of associations between genetic variation and common diseases. A Network of Investigator Networks has been set up to drive the process, sponsored by the Human Genome Epidemiology Network. A workshop is planned to develop consensus guidelines for reporting results of genetic association studies. Published literature databases will be integrated, and unpublished data, including 'negative' studies, will be captured by online journals and through investigator networks. Systematic reviews will be expanded to include more meta analyses of individual-level data and prospective meta-analyses. Field synopses will offer regularly updated overviews. PMID- 16468122 TI - Common deletion polymorphisms in the human genome. AB - The locations and properties of common deletion variants in the human genome are largely unknown. We describe a systematic method for using dense SNP genotype data to discover deletions and its application to data from the International HapMap Consortium to characterize and catalogue segregating deletion variants across the human genome. We identified 541 deletion variants (94% novel) ranging from 1 kb to 745 kb in size; 278 of these variants were observed in multiple, unrelated individuals, 120 in the homozygous state. The coding exons of ten expressed genes were found to be commonly deleted, including multiple genes with roles in sex steroid metabolism, olfaction and drug response. These common deletion polymorphisms typically represent ancestral mutations that are in linkage disequilibrium with nearby SNPs, meaning that their association to disease can often be evaluated in the course of SNP-based whole-genome association studies. PMID- 16468123 TI - New goals for contrast-enhanced sonography. PMID- 16468124 TI - Frequency modulation of motor unit discharge has task-dependent effects on fluctuations in motor output. AB - The rate of change in the fluctuations in motor output differs during the performance of fatiguing contractions that involve different types of loads. The purpose of this study was to examine the contribution of frequency modulation of motor unit discharge to the fluctuations in the motor output during sustained contractions with the force and position tasks. In separate tests with the upper arm vertical and the elbow flexed to 1.57 rad, the seated subjects maintained either a constant upward force at the wrist (force task) or a constant elbow angle (position task). The force and position tasks were performed in random order at a target force equal to 3.6 +/- 2.1% (mean +/- SD) of the maximal voluntary contraction (MVC) force above the recruitment threshold of an isolated motor unit from the biceps brachii. Each subject maintained the two tasks for an identical duration (161 +/-93 s) at a mean target force of 22.4 +/-13.6% MVC. As expected, the rate of increase in the fluctuations in motor output (force task: SD for detrended force; position task: SD for vertical acceleration) was greater for the position task than the force task (P < 0.001). The amplitude of the coefficient of variation (CV) and the power spectra for motor unit discharge were similar between tasks (P > 0.1) and did not change with time (P > 0.1), and could not explain the different rates of increase in motor output fluctuations for the two tasks. Nonetheless, frequency modulation of motor unit discharge differed during the two tasks and predicted (P < 0.001) both the CV for discharge rate (force task: 1-3, 12-13, and 14-15 Hz; position task: 0-1, and 1-2 Hz) and the fluctuations in motor output (force task: 5-6, 9-10, 12-13, and 14-15 Hz; position task: 6-7, 14-15, 17-19, 20-21, and 23-24 Hz). Frequency modulation of motor unit discharge rate differed for the force and position tasks and influenced the ability to sustain steady contractions. PMID- 16468125 TI - Re: An unusual complication of interscalene brachial plexus catheterization: delayed catheter migration. PMID- 16468126 TI - Re: Near miss drug administration. PMID- 16468127 TI - Counterpoint: in health and in normoxic environment VO2max is limited primarily by cardiac output and locomotor muscle blood flow. PMID- 16468128 TI - Comment on Point:Counterpoint "The muscle metaboreflex does/does not restore blood flow to contracting muscles". PMID- 16468129 TI - Comment on Point:Counterpoint "The muscle metaboreflex does/does not restore blood flow to contracting muscles". PMID- 16468130 TI - [Ultrasonography of the kidney and renal vessels. I. Normal findings, inherited and parenchymal diseases]. AB - Renal ultrasonography has become the standard imaging modality in the investigation of kidneys because it displays excellent anatomic detail, requires no special preparation of the patient and does not expose the patient to radiation or contrast agents. Ultrasonography is used to determine the site and size of the kidney and to detect local lesions like tumors, cysts and renal stones. Furthermore the presence and urodynamic relevance of hydronephrosis can reliably be revealed. Also reno-parenchymatous diseases are discernible to the experienced investigator, however most glomerular diseases cannot be further subclassified. Exceptions are primarily renovascular disorders like hypertensive nephrosclerosis, diabetic nephropathy or renal vasculitis. Color Doppler sonography allows the detection and quantification of renal artery stenosis, increased resistance index values may indicate irreversible disease. Ultrasonography has also been found of value in the evaluation of renal transplant kidneys. Especially in the early transplant course potentially fatal but reversible diseases like renal vein thrombosis or urinomas are detected with high sensitivity. In the long term, an increased resistance index value may also predict allograft failure. PMID- 16468131 TI - [D-dimer screening for clinical recognition of silent thrombosis]. PMID- 16468132 TI - [Vascular access in emergency pediatric anesthesia]. PMID- 16468133 TI - [Vascular access in pediatric anesthesia]. PMID- 16468134 TI - [Comments on 'The treatment of sialorrhea with botulinum toxin']. PMID- 16468135 TI - Gastro-colonic anastomosis--a viable option in extensive small bowel infarction. AB - INTRODUCTION: We have previously presented a patient with massive small and large bowel infarction and demonstrated that even with only a few inches of remaining small bowel an almost normal life-style and diet is possible. PATIENT: Recently, we have looked after a young and otherwise fit female patient who suffered mesenteric venous gangrene of the whole small bowel from the Ligament of Treitz to the caecum. In order to achieve gastro-intestinal continuity and to avoid the torrential fluid loss associated with high fistula, an anastomosis between the stomach and the transverse colon was formed. RESULTS: We are surprised to find that despite the extensive resection our patient maintains a good quality of life and is able to look after her young family. PMID- 16468136 TI - Laparascopic access: a different approach. PMID- 16468137 TI - Antegrade porridge enema to assess anorectal function after severe perineal sepsis. PMID- 16468138 TI - Use of a 'mirror plate' in minimally invasive plate osteosynthesis. PMID- 16468139 TI - A simple surgical technique for removal of radio-opaque foreign objects from the plantar surface of the foot. PMID- 16468140 TI - A technique to assist reversal of abdominal stomas. PMID- 16468141 TI - Vascular clamping of calcified vessels: the peanut technique. PMID- 16468142 TI - Percutaneous tibial plate removal. PMID- 16468143 TI - Easy placement of transanastomotic nasojejunal triple lumen feeding tube during gastrectomy. PMID- 16468144 TI - Optimum incision length for port insertion in laparoscopic surgery. PMID- 16468145 TI - Arthroscopically-assisted percutaneous fixation of a posterior fracture dislocation of the proximal humerus. PMID- 16468146 TI - Cementing technique made easy. PMID- 16468147 TI - [Autoimmune thyroid diseases and associated diseases. Continuing medical education seminar on endocrinology and immunology. 15 June 2005, Belgrade]. PMID- 16468148 TI - Letter regarding article by Norman et al, "novel mutation in desmoplakin causes arrhythmogenic left ventricular cardiomyopathy". PMID- 16468149 TI - Does the European pharmaceutical industry have a future? PMID- 16468150 TI - A new Hungarian revolution. PMID- 16468151 TI - Baby boomers could be the straw that breaks LTC's back. PMID- 16468152 TI - [Patterns of hearing disorders in normal otoacoustic emissions]. AB - OBJECTIVE: To analyze and outline patterns of hearing disorders in normal otoacoustic emissions. METHOD: Tests of pure tone audiometry, acoustic impedance test, auditory brainstem response (ABR), 40 Hz AERP,spontaneous otoacoustic emissions (SOAE), transient evoked otoacoustic emissions (TEOAE), distortion evoked otoacoustic emissions (DPEOAE)and imaging examinations of CT/MRI scan were performed on all 83 patients with hearing disorders. RESULT: Patterns of hearing disorders in normal otoacoustic emissions including (1) 68 patients with auditory neuropathy; (2) 2 patients with unilateral acoustic neuroma; (3) 3 patients with cortical deafness or central hearing loss; (4) 2 patients with hyperacusis; (5) 2 patients with functional deafness; (6) 6 patients with malingering. CONCLUSIONS: OAE tests play an important role in diagnosis and differential diagnosis of the patients with retrocochlear deafness, central hearing loss, nonorganic hearing loss and some other special hearing disorders. PMID- 16468153 TI - Gallstones. Common, but not to be ignored. PMID- 16468154 TI - Health tips. Strength training provides benefits. PMID- 16468155 TI - Device may reduce stroke risk from chronic atrial fibrillation. PMID- 16468156 TI - Bird flu. Surviving a pandemic. PMID- 16468157 TI - Beyond Viagra. What are your options? PMID- 16468158 TI - I've decided to reduce the amount of meat I eat to improve my health and lose weight. In place of meat, I'd like to switch to meat substitutes. Any recommendations? PMID- 16468159 TI - Evidence that the Vinland Map is medieval. AB - The Vinland Map has been proposed to date to the mid-fifteenth century and is significant in showing the north Atlantic area of what is now North America about 50 years before the voyages of Columbus. It has been confirmed by radiocarbon dating that the map was drawn on a fifteenth century parchment; however, controversy regarding the ink of the map has persisted. Analytical studies of the ink of the map have been interpreted to show that the map is a twentieth century forgery because of the presence of carbon and anatase in the ink. However, carbon is known to have been added to medieval iron gallotannate inks, and the presence of anatase in a medieval ink can be explained by examining the process of making medieval inks. PMID- 16468161 TI - Proceedings of the LVIII Annual Meeting of the Italian Society for Veterinary Science (SISVET). 2004. Grado, Italy. PMID- 16468160 TI - Side effects associated with the treatment of multidrug-resistant tuberculosis. AB - SETTING: Sureyyapasa Centre for Chest Diseases and Thoracic Surgery, Istanbul, Turkey. OBJECTIVE: To report the frequency of treatment side effects in cases of multidrug-resistant (MDR-TB) tuberculosis. DESIGN: A retrospective review of the medical records of 263 patients who received individualised treatment for MDR-TB between April 1992 and June 2004. RESULTS: One or more side effects developed in 182 cases (69.2%). These effects led the clinicians to withdraw one or more drugs from the treatment regimen in 146 cases (55.5%). Side effects observed most frequently included: ototoxicity (41.8%), psychiatric disorders (21.3%), gastrointestinal disturbance (14.0%), arthralgia (11.4%), epileptic seizures (9.9%), hepatitis (4.5%), and dermatological effects (4.5%). At the time of analysis, treatment was successful in 204 (77.6%) cases. Fifty-nine patients (22.4%) had poor outcomes. CONCLUSION: Timely and aggressive management of drug side effects means that high side effect rates in MDR-TB treatment need not compromise success rates. PMID- 16468162 TI - Iodixanol and chronic kidney disease. PMID- 16468164 TI - Proceedings of the 7th Jenner Glycobiology and Medicine Symposium. September 5-8, 2004. PMID- 16468163 TI - Cytokines as potential therapeutic targets for inflammatory skin diseases. Proceedings of a workshop. November 17-19, 2004. Napa Valley, California, USA. PMID- 16468166 TI - Proceedings of the International Symposium on Uncertainty and Precaution in Environmental Management. June 7-9, 2004. Copenhagen, Denmark. PMID- 16468165 TI - New aspects in gynecologic oncology. PMID- 16468167 TI - [Management of the documentation and registries of the radiopharmacy units. General texts of the Royal Spanish Pharmacopeia with character of recommendation]. PMID- 16468168 TI - Proceedings of the 12th European Conference of the Pharmacochemistry Grouping of the Atlantic Arc (GP2A). 2003. La Rochelle, France. PMID- 16468169 TI - A critical assessment of text mining methods in molecular biology. Proceedings of a workshop. March 28-31, 2004. Granada, Spain. PMID- 16468170 TI - Birth and name-giving in Hittite texts. PMID- 16468171 TI - Gleanings from the Iraq medical journals. PMID- 16468172 TI - The double life of lentiviruses. PMID- 16468174 TI - Resetting the clock. PMID- 16468173 TI - Tiny droplets make a big splash. PMID- 16468176 TI - Pinning the tail on the substrate. PMID- 16468175 TI - A new avenue for mass spectrometry. PMID- 16468177 TI - Training GFP to fold. PMID- 16468178 TI - Animal husbandry at Uruk in the Archaic Period. PMID- 16468179 TI - A synthetic classic reinvented. PMID- 16468180 TI - Reliability and validity of observer ratings of neighborhoods. AB - OBJECTIVES: The objectives were to examine the validity and reliability of a five item neighborhood measurement scale. METHODS: Respondents were enrolled from two catchment areas: a poor inner city and a heterogeneous suburban area. Items combine for a total score of 5 (best) to 20 (worst). The authors compared scales across catchment areas and respondent ratings and assessed interviewer effects and retest reliability. RESULTS: Suburban neighborhood scale scores were 3 points lower (higher socioeconomic status, SES) than the inner-city scores. There was a strong relationship between scores and participants' neighborhood ratings. The retest correlation was substantial (.81), but only two of five items achieved kappas above .75. In regression models, interviewer experience and residence and individual interviewers contributed to different ratings, although there was still a marked difference between catchment areas. DISCUSSION: Observer ratings of neighborhoods show promise as a measure of neighborhood SES, despite problems with interviewer effects. Future work should improve objective criteria for ratings. PMID- 16468182 TI - Intraperitoneal treatment lengthens survival in ovarian cancer. PMID- 16468181 TI - New hope for old drug in patients with brain metastases. PMID- 16468183 TI - Genetic testing for breast cancer varies across UK. PMID- 16468184 TI - Researchers find Singapore difficult to resist. PMID- 16468185 TI - Stamping out blindness. PMID- 16468186 TI - Myths, magic and medicine. PMID- 16468188 TI - Forging a new direction--WSU. PMID- 16468187 TI - Ethnic tensions aggravate nursing shortage. PMID- 16468189 TI - Dilmun: gateway to immortality. PMID- 16468190 TI - Omen texts concerning lesser known parts of the lungs. PMID- 16468191 TI - Settlement surveys and documentary evidence: regional variation and secular trend in Mesopotamian demography. PMID- 16468192 TI - A uterine amulet in the Oriental Institute collection. PMID- 16468193 TI - Another old Babylonian childbirth incantation. PMID- 16468194 TI - A short biography of the life of the dedicated anatomist - Valsalva. PMID- 16468195 TI - Echoes of a thirsty owl: death and afterlife in Pre-Islamic Arabic poetry. PMID- 16468196 TI - Blindness and night-blindness in Akkadian. PMID- 16468197 TI - The natural history of the Tigris according to the Sumerian literary composition Lugal. PMID- 16468198 TI - Urbanization and the organization of animal production at Tell Jemmeh in the Middle Bronze Age Levant. PMID- 16468199 TI - The evil eye in Mesopotamia. PMID- 16468200 TI - Northwest Semitic incantations in an Egyptian medical papyrus of the fourteenth century B. C. E. PMID- 16468201 TI - Materials on Averroes's circle. PMID- 16468203 TI - The samanu disease in Babylonian medicine. PMID- 16468202 TI - The purpose of the Parthian Galvanic cells: a first-century A.D. electric battery used for analgesia. PMID- 16468204 TI - Innovations and adaptations in ancient Egyptian medicine. PMID- 16468205 TI - The scorpion spell from Wadi Hammamat: another Aramaic text in Demotic script. PMID- 16468206 TI - Ethnography and archaeological interpretation of funerary remains. PMID- 16468207 TI - Socio-political implications of lowland Maya burials: methodology and tentative hypotheses. PMID- 16468208 TI - Pleistocene human remains from Australia: a living site and human cremation from Lake Mungo, Western New South Wales. PMID- 16468209 TI - Pleistocene man in South America. PMID- 16468210 TI - The diet of early man: aspects of archaeological evidence from Lower and Middle Pleistocene Sites in Africa. PMID- 16468211 TI - Physical evolution and technological evolution in man: a parallelism. PMID- 16468212 TI - Forensic aspects of the so-called Neolithic skeleton Q1 from Maiden Castle, Dorset. PMID- 16468213 TI - Two static maximum population-density models for hunter-gatherers: a first approximation. PMID- 16468214 TI - Population, economy and society in the Chassey-Cortaillod-Lagozza cultures. PMID- 16468215 TI - Palaeodemography and earlier British populations. PMID- 16468216 TI - Ecology and population in the Eastern Mediterranean. PMID- 16468217 TI - Population trends in the Teotihuacan Valley, A.D. 1400-1969. PMID- 16468219 TI - Systemic population control in the Middle and Upper Palaeolithic: inferences based on contemporary hunter-gatherers. PMID- 16468218 TI - Population control among hunter/gatherers. PMID- 16468220 TI - Hunter-gatherer populations and archaeological inference. PMID- 16468221 TI - Animal bone finds and economic archaeology: a critical study of 'osteo archaeological' method. PMID- 16468222 TI - Environment and subsistence of prehistoric man in the southern Cape Province, South Africa. PMID- 16468224 TI - Further refinement of formulae for determining population from floor area. PMID- 16468227 TI - Two weeks in Banda Aceh. PMID- 16468226 TI - Ovarian cancer. AB - Ovarian cancer is the leading cause of death from gynecologic cancers. Although approximately 2,400 women are diagnosed each year in Canada, most women know little about the disease. While it is not common, ovarian cancer is a devastating diagnosis. The symptoms of ovarian cancer are non-specific and there is no effective screening tool. Surgery, chemotherapy and radiation given alone or in combination are most commonly used to treat ovarian cancer. Surgery can result in premature menopause for women in their child-bearing years. Chemotherapy may cause a multiplicity of distressing side-effects. Women with ovarian cancer are living longer and with an improved quality of life. Nurses are in a key position to increase awareness and provide support and information about coping with the disease and side-effects of treatment. PMID- 16468228 TI - A noble vocation. Interview by Barbara Sibbald. PMID- 16468229 TI - Thyroid tumors: histological classification and genetic factors involved in the development of thyroid cancer. AB - Classification of thyroid tumours and their variants is described with special respect to some recent findings on somatic mutations characteristics which are associated with individual types of malignity. Special attention is paid to the interrelations between thyroid nodules and malignity and predictive risk factors are listed. PMID- 16468230 TI - Prospective analysis of 518 cases with thyroidectomy in Turkey. AB - OBJECTIVE: Thyroid malignancies can present in different manners, among them as asymptomatic solid nodule being the most puzzling. Nodules have been found in the 60-70 % of autopsy specimens and it is very important to rule out the malignancies in such cases. Incidence of carcinomatous changes is reported in 5 15 % of solitary nodules. We present the results of prospective study on 418 thyroidectomies with the aim to review the experience of our unit, to establish the correlation between clinical presentation and histopathology, to discuss the malignancy rates and surgical complications. PATIENTS AND METHODS: Five hundred eighteen consecutive cases of thyroidectomy 419 female (80.8 %), and 99 male (19.2 %) patients performed between January 2002 and October 2004 were included in this prospective study. RESULTS: In 71 (13.7 %) cases the malignancy was found by paraffin specimens, the highest prevalence of malignancy being found in patients with nodular goiter (NG-18 %) followed by 14.6 % in multinodular goiter (MNG). The sensitivity of preoperative fine needle cytology (FNAC) was 83.3 % with false positive rate of 1.3 %. Complications were seen in 5.2 % of cases of which 4 (0.7 %) had hypoparathyroidism and 7 (1.3 %) had recurrent laryngeal nerve injury. All patients observed came from endemic area. Family history nearly doubles the risk of malignancy. CONCLUSIONS: In an endemic area the nodular goiter is the most common. Preoperative cytology, although sensitive, gives a considerable number of false positive results. Results of thyroid surgery at a high volume centre are satisfactory with very low rates of recurrent laryngeal nerve and parathyroid injury. Probability of malignant transformation in a long standing thyroid swelling should always be kept in mind. There appears to be an increase in prevalence of thyroid malignancies in Turkey after Chernobyl disaster. PMID- 16468231 TI - Clinical presentation and treatment of hyperthyroidism associated with thyroid cancer. AB - OBJECTIVE: Hyperthyroidism apparently does not protect the patients from thyroid cancer as believed before. In contrast, hyperthyroidism with concurrent thyroid cancer can be diagnosed after pathological examination of unsuspect nodules. The aim of this study was to evaluate the coexistence of hyperthyroidism and thyroid carcinoma and to discuss the advantages of total thyroidectomy in such cases. METHODS: Between January 2002 and October 2004, 120 hyperthyroid patients underwent surgical treatment in our clinic. All patients with hyperthyroidism in this study underwent fine-needle aspiration biopsy and cytologic examination. Frozen section evaluation was performed in all of these patients during the operation. RESULTS: Among these patients 10 had concurrent thyroid cancer. Only one of these patients was examined by fine needle aspiration biopsy prior to operation, while the rest of malignancies was diagnosed from unsuspect nodules. CONCLUSIONS: The selection of appropriate operation procedure appears very important to find out and treat concurrent thyroid cancers. We diagnosed 90 % of thyroid cancers incidentally. If there are no technical difficulties, we prefer total thyroidectomy for the patients with toxic multinodular goiter and Graves' disease with nodules. PMID- 16468232 TI - Oxytocin--anatomy and functional assignments: a minireview. AB - Oxytocin (OXY) is a very abundant neuropeptide exerting a wide spectrum of central and peripheral effects as neurohormone, neurotransmitter, or neuromodulator. In the central nervous system (CNS), the OXY gene is predominantly expressed in magnocellular neurons in the hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei. The magnocellular OXY neurons release their products into the general circulation in the neurohypophysis while the mediocellular OXY neurons secrete elsewhere in the CNS. OXY is also produced in peripheral tissues, e.g., uterus, placenta, amnion, corpus luteum, testis, and heart. OXY is a potent stimulator of spontaneous erections in rats and is involved in ejaculation. The typical actions of peripheral OXY are stimulation of uterine smooth muscle contraction during labor and milk ejection during lactation. OXY acts via the receptor which is a typical class of I G protein coupled receptor. OXY receptors have also been identified in other tissues, including the kidney, heart, thymus, pancreas, and adipocytes. PMID- 16468234 TI - [Some new findings in the pathogenesis of myeloproliferative disorders and new insight into more effective treatment]. AB - Advances in molecular biology and genomics enabled more detailed view on the pathogenesis of myeloproliferative disorders, which are considered to be a diseases of hematopoietic stem cell. The autor provides an brief overview of the genetic alterations, leading to the chronic myeloid leukemia, myelodysplastic syndrome and acute myeloid leukemia. In the second part, molecular targeted therapies that have been developed based on these insights are reviewed. These are particularly methods preventing increased proliferation such as inhibition of tyrosinkinases (imatinib, dasatinib), inhibitions of farnesyltransferase (tipifarnib), inhibition of angiogenesis (bevacizumab, vatalanib), induction of diferentiation (hypomethylating agenents) and induction of apoptosis (bortezomib). More detailed information is given on some novel drugs which are currently in clinical trials. PMID- 16468235 TI - [Principle and importance of using the array CGH in hematooncology]. AB - Identification of chromosomal changes and variation in DNA copy number allows us to understand pathogenesis of tumors. To the frequently diagnosed chromosomal changes belong acquired gains and losses of chromosomal regions carring genes involved in cellular proliferation and differentiation as well as oncogenes and tumor suppressor genes. The determination of gene changes is limited by techniques used for their identification. The introduction of genom-wide microarray technology, resolution has rapidly increased. Array comparative genomic hybridization (arrayCGH) offers higher resolution for genome-wide detection of chromosomal alteration and it is able to analyze hundreds to thousands of genes presented on microarray in one experiment. The aim of this study was to perform arrayCGH technology and to stress its value for the identification of chromosomal imbalances in hematological malignancies. PMID- 16468236 TI - [Acylation stimulating protein--its role in control of metabolism in the adipose tissue]. AB - Multiple hormones and enzymes participate in the lipid storage. One of them is acylation stimulating protein. Acylation stimulating protein is produced predominantly by adipocytes. After the binding on specific receptor at the surface of adipocytes, acylation stimulating protein leads to enhancement of triglyceride synthesis. This process is mediated by protein-kinase C. Concurrently, glucose transporters move from the cytoplasm to the adipocyte surface. Higher glucose disposal leads to a sufficient substrate availability for triglyceride synthesis. Acylation stimulating protein also stimulates pancreatic insulin secretion. Total acylation stimulating protein level in plasma is related to the adipose tissue mass and it positively correlates with many anthropometric parameters and with serum insulin level. In acylation stimulating protein deficient mice, resistance to the obesity development after a high fat diet was observed. Adipose tissue mass is lower in the acylation stimulating protein deficient mice and higher insulin sensitivity was shown in acylation stimulating protein deficient mice compared to a wild type mice. Acylation stimulating protein pathway may have an important role in the obesity development. PMID- 16468237 TI - [High dose therapy with autologous stem cell transplantation in patients with Hodgkin's lymphoma: long-term follow-up in patients treated in one center]. AB - BACKGROUND: Majority of patients with Hodgkin's Lymphoma (HL) can be cured by first line therapy. The high dose therapy (HDT) with autologous stem cell transplantation (ASCT) is the option which can be used in the situation when the conventional therapy failed. METHODS AND RESULTS. Beginning 1994 till 2005 84 pts with HL who did not respond the conventional chemotherapy underwent 105 HDT procedures with ASCT. The median age at the time of HDT was 30.5 years. The reason for salvage therapy followed by HDT with ASCT was the failure to achieve 1st complete remission-- CR (n 16) or the subsequent relapse or progression (n 68). The disease status at the time of HDT after conventional salvage chemotherapy was assessed as chemosensitive in 65 pts (77.4%) and chemoresistant in 19 pts (22.6%). The most frequent HDT regimen used was BEAM (82 HDT), 22 pts entered into the tandem HDT program. Bone marrow only was used as the source of progenitor cells in 4 ASCT, peripheral blood progenitor cells (PBPC) only were used in 85 ASCT and the combination of both in 16 ASCT. The disease status after the HDT with ASCT was assessed (77 pts were qualifiable) as CR in 39 pts (50.6%), PR in 31 (40.3%) and as stable disease or progression in 7 pts (9.1%). Treatment related mortality in HDT with PBPC was 3.9%. The median follow up is 5.3 years. The five year probability of event free survival (EFS) is 43.1% and overall survival 53.2%. The EFS and OS probability respectively for the chemosensitive patients was 48.6% and 62.9% respectively. The status at HDT and the results after it have prognostic significance. There were observed 39 deaths and 26 of them were caused by disease progression. Secondary tumor was observed in 5 pts and in all of them it caused the death. CONCLUSIONS: The HDT with ASCT allows the long-term survival without disease progression in about a half of the patients with reasonable toxicity. PMID- 16468238 TI - [Importance of quantitative evaluation of BCR-ABL transcripts using real-time PCR for effective treatment of chronic myeloid leukemia]. AB - BACKGROUND: Molecular biology methods based on reverse transcription and polymerase chain reaction (RT-PCR) are able to detect the presence of BCR-ABL transcripts in chronic myeloid leukemia (CML). In this study we present our experience with monitoring of residual disease using real-time PCR with hybridization probes detection in patients treated with imatinib mesylate and in collected peripheral blood progenitor cells (PBPC). METHODS AND RESULTS: We measured the level of BCR-ABL transcripts in peripheral blood cells of 27 subjects before and in the course of the imatinib treatment. The median of relative quantity of BCR-ABL in the blood before imatinib therapy was 2.55%. The number of the transcripts in 23 imatinib-sensitive subjects decreased to 0.02% in 6 months. After 12 months of the treatment the BCR-ABL median was 0.005%. Subsequent levels fluctuated between values below the detection limit (DL, 0.001%) and 0.005%. Three patients were primarily resistant to imatinib with the BCR-ABL range of 0.13%-11.7% during the treatment. One subject showed marks of molecular relapse after 18 months of the treatment. Only two of 16 filgrastim stimulated patients had BCR-ABL levels in the blood and in collected PBPC below DL. In other subjects BCR-ABL transcripts were determined within the measurable range of RT-PCR. CONCLUSIONS: Taking into account prognostic importance, the measurement of BCR-ABL transcripts is an effective approach to monitoring of residual CML kinetics. Evaluation of BCR-ABL levels in collected PBPC can complete information on quality of the cells in potential autotransplants, and choose subsequent therapeutic protocols and patient prognosis. PMID- 16468239 TI - [Significance of confirmatory testing of HLA system in unrelated haematopoietic stem cell donors]. AB - BACKGROUND: Haematopoietic stem cell transplantation is a standard curative therapy for some acquired haematological diseases and inherited metabolic and immunological disorders. The HLA compatibility in five loci (HLA class I -A, -B and -C and HLA class II -DRB1 and -DQB 1) of the donor/recipient pair is a prerequisite for the success of haematopoietic stem cell transplantation which represents a process of adoption donors immunity. METHODS AND RESULTS: HLA is the most polymorphic system in the human genome and this polymorphism is exactly detected by molecular genetics methods on DNA level only. In period of 2001-2004 we performed confirmatory testing of 366 unrelated haematopoietic stem cells donors from Czech and foreign registers for 256 patients. Only 16% of the donors completely matched the patients in all HLA loci. We detected HLA mismatches in the samples of 81% patient/donor pairs but these results were consonant with previous results from registers. 3% of confirmatory samples were discrepant with previous registry data. CONCLUSIONS: Despite of increasing number of available unrelated haematopoietic stem cell donors and the quality of registry HLA typing the possibility of finding the completely match donor is still limited. PMID- 16468240 TI - [The use of quantitative assessment of Wilms tumour gene 1 for monitoring of residual disease in acute myeloid leukemia patients]. AB - BACKGROUND: Despite a considerable effort, the majority of acute myeloid leukaemia (AML) patients do not have a suitable specific molecular marker for monitoring minimal residual disease (MRD). The results of some studies suggest the Wilms tumour gene (WT1) as a possible molecular marker of MRD. METHODS AND RESULTS: We measured the expression of WT1 at diagnosis and during treatment of the acute myeloid leukaemia (AML) patients. The expression of WT1 was measured by the quantitative real-time RT-PCR in peripheral leukocytes from 56 AML at diagnosis and 7 patients with AML transformed from myelodysplastic syndromes (MDS). The WT1 expression was significantly elevated (up to 3 orders of magnitude) in peripheral blood samples (PB) of AML patients at diagnosis compared to PB samples of healthy donors (P < 0.0001). The level of WT1 expression depends particularly on FAB AML subtype, with the highest being found in AML patients with subtypes M4, M1, M3 and AML transformed from MDS. Conversely, AML patients with M2 and with the presence of AML1/ET0 at presentation showed a significantly lower expression of the WT1 gene compared to the remaining AML patients at presentation (P = 0,005). Further, sequence samples of 12 AML patients under long term surveillance were tested for the WT1 expression in parallel with the expression of specific MRD markers--fusion genes: AMLI/ETO, PML/RARalpha and CBFB/MYH11. The levels of WT1 gene expression and the above specific fusion genes significantly correlated. Moreover, 14 patients without the specific MRD marker were tested for the WT1 expression. The results show that haematological relapses were associated with the rise of expression of the specific fusion genes and with the WT1 gene expression. The rise of WT1 expression above the level seen in leucocytes from peripheral blood and/or bone marrow of healthy donors--in four patients under long-term surveillance the "molecular relapse" predicted ongoing haematological relapses as early as 2 months in advance. CONCLUSIONS: Our results, in accordance with some of the previously published ones, show that WT1 expression seems to be a suitable marker of minimal residual disease in AML patients. PMID- 16468241 TI - [Acute adverse effects of high dose brachytherapy in combination with external radiotherapy in localized prostate cancer]. AB - BACKGROUND: Over the past few years, brachytherapy has become more and more common in the treatment of prostate cancer. Part of the reason behind this growth in the use of brachytherapy for the treatment of organ-confined prostate cancer is the reduced amount of acute and chronic side effects. On the other hand, brachytherapy allows for dose escalation, resulting in significant improvements in the treatment results. METHOD AND RESULTS: From August 2004 to June 2005, we irradiated 40 patients with T1c-T3a prostate cancer. All of the patients underwent transperineal transrectal guided high dose rate (HDR) brachytherapy (two fractions, 8 Gy per fraction) and course of external beam radiotherapy with a median dose of 45-50.4 Gy. The patients were classified into three groups: low risk of recurrence (11 patients - 27.5%), medium risk (14 patients - 35%) and high risk (15 patients - 37.5%). The medium age of the patients was 68.7 years (range of between 55 and 77). Hormonal treatment was carried out with 17 of the patients (42.5%). We evaluated the quality of each implantation, including the maximum urethral and rectal dose. The calculated doses were compared with measurements by in vivo dosimetry. Acute toxicity was evaluated in all of the patients according to the Radiation Therapy Oncology Group (RTOG) scale. Each of the patients completed an International Prostatic Symptom Score (IPSS) questionnaire. Acute genitourinary morbidity grade 1 was recorded in 37.5% of patients, and grade 2 in 15% of patients. Urine retention in one of the patients resulted in the need to perform an epicystostomy. According to the IPSS score, the majority of patients (90%) experienced an improvement in symptoms related to quality of life. Grade 1 acute gastrointestinal toxicity was recorded in 40% of the patients. Grades 2-4 were not recorded. CONCLUSIONS: We showed that the combination of external beam radiotherapy and HDR brachytherapy in the treatment of early prostate cancer to be feasible and well tolerated. Acute toxicity was low and scarcely influenced the quality of life. Among the risk factors of genitourinary toxicity was the volume of the prostate. For gastrointestinal toxicity, risk factors included a combination of HDR brachytherapy and external beam radiotherapy to the pelvis, as well as hormonal treatment. PMID- 16468242 TI - [Familial haemophagocytic lymphohistiocytosis caused by perforin deficit can be successfully treated by haematopoietic stem cell transplantation--the first diagnosed case in the Czech Republic]. AB - Familial haemophagocytic lymphohistiocytosis (FHL) is an inherited disorder characterized by an impaired cytotoxicity of T lymphocytes and NK cells typically manifesting within first few months after birth. If not treated adequately, it is inevitably fatal within several months. The incidence in Caucasians has been estimated to 1: 50 000 births. Haematopoietic stem cell transplantation represents the only curative treatment for FHL. Recently, several genetic defects underlying molecular defects in FHL have been identified. In approximately 30% of patients FHL is caused by mutations in PRF1 gene coding for perforin. Further 30% of patients were found to have mutations in UNC13D coding for hMunc13-4 protein. Very recent report has identified another cause of FHL, mutations in STX11 gene on chromosome 6, coding for syntaxin 11. Absence of any of those proteins severely impairs the process of exocytosis of cytotoxic granules. We describe patient with clinical symptoms of FHL. Immunological and molecular biology methods led to the identification of perforin mutation as a cause of the disease. Patient received an allogeneic SCT from HLA-matched unrelated donor. SCT was followed by rapid normalization of clinical symptoms and laboratory findings. In patient described in this study, FHL manifested with typical clinical and laboratory symptoms. Adequate immunosuppressive treatment and subsequent SCT led to the sustained remission of FHL and correction of molecular defect. This is the first case of FHL in Czech Republic where perforin mutation was identified as a molecular cause both at cellular and molecular level. PMID- 16468243 TI - [Hydatic cysts--echinococcosis]. AB - The case diagnosis and treatment of hydatic cyst in the right and left lobe of the liver in a 28 year old pregnant woman is described in the article. PMID- 16468244 TI - [Abuse of buprenorphine becomes a problem of the Czech Republic]. AB - Buprenorphine (Subutex) is according to epidemiological data and clinical experience abused on a large scale in the Czech Republic and for some drug dependent persons it becomes a principal intravenously applied drug. This problem can be resolved by more appropriate training of physicians who prescribe the drug and especially by the introduction of the combination buprenorphine and naloxone (Suboxone) which is not abused intravenously. The registration of Suboxone in the Czech Republic was not initiated by the manufactures so far. PMID- 16468245 TI - [Robotic surgery]. AB - Installation first robotic system DaVinci in Prague in Czech Republic brought many questions. Surgery has rapidly changed during last 150 years. The progress in endoscopic surgery concerned namely the general surgery, thoratic and cardiovascular surgery. Present article gives and overview on the development of surgical techniques in the world from the laparoscopic to robotic surgery. PMID- 16468246 TI - [Impact factor--good servant, but a bad master]. AB - Our paper presents an overview on the role of impact factor (IF) in the valuation of the scientific activity. With maximal objectivity based on published articles the perspectives are outlined and the role of specialized medical journals like Journal of Czech Physicians is shown. When IF is used for evaluation, the full English term should be first considered "journal impact factor". It expresses the bibliometric features of the given journal, without possibility to transfer this assessment to individual articles or even authors. IF determined on the basis of information from citation indexes represents for an article presented in the journal the probability to be cited. It appears to be evident that this valuable parameter can lead in hands of inexperienced or irresponsible evaluator to incorrect and erroneous conclusions or to be misused. It is useful neither for evaluation of publication or scientific activity of an individual nor for the comparison of journals in different scientific disciplines. Owing to the relation of IF to the probability of an article to be cited, IF reflects the features (quality) of an article which brought the citation. IF is therefore determined also by the activity of scientific editors and referees. IF becomes part of everything what is related to the publication of scientific results and therefore it becomes object of journalogy. It is the source of meritorious intentions of the editorial board of Journal of Czech Physicians to deal with this phenomenon. PMID- 16468247 TI - [The most eminent personalities of the Prague Medical Faculty. Edward Babak]. PMID- 16468248 TI - Death: "the distinguished thing". PMID- 16468249 TI - The end of autonomy. PMID- 16468250 TI - Living long in fragile health: the new demographics shape end of life care. PMID- 16468251 TI - Conflict and consensus at the end of life. PMID- 16468252 TI - Hope for the future: achieving the original intent of advance directives. PMID- 16468253 TI - Recognizing death while affirming life: can end of life reform uphold a disabled person's interest in continued life? PMID- 16468254 TI - Making room for dying: end of life care in nursing homes. PMID- 16468255 TI - The past and future of palliative care. PMID- 16468256 TI - The role of litigation in end of life care: a reappraisal. PMID- 16468257 TI - The quest to reform end of life care: rethinking assumptions and setting new directions. PMID- 16468259 TI - [Health education with community groups: a strategy that facilitates health promotion]. AB - This paper highlights a few aspects of health promotion and education that enable reasoning over the pertinence of the work developed with groups in the community. Group work makes it possible breaking the traditional vertical relationship between the health professional and the subject of his/her action. It is also a strategy that facilitates the expression of individual and collective needs, expectations and life circumstances that influence health. PMID- 16468260 TI - [Talking about research, education and health in nursing]. AB - The objective of this paper is making some theoretical reflections about the significance of research and education to the construction of knowledge and their influence in the health-disease process. It is understood that the materialization of educative actions, to be developed by the graduated nurse, requires a scientific base,founded on research and interactive with the environment where the human being is inserted. PMID- 16468261 TI - [Gadamer: grounding a proposal for the study of stress in the surgical center]. AB - This article makes a reflection on philosophical foundation to support the methodology in order to study the stress of professionals who work in the operating theater. The qualitative research allows a comprehensive access to the world of each subject, revealing the individual perception of whom experiences the phenomenon. Gadamer understands that the language reveals and is suitable to evaluate properly, through hermeneutics, the symbolic dimension of the subject's speech. Hermeneutics is not only just a theory of the comprehension and interpretation art but much more than that, it is close to the practical philosophy, having to respond for the methodological feature of things whose understanding it provides. PMID- 16468262 TI - [Accidents with biological material at West Parana University Hospital]. AB - It is a descriptive and retrospective study with the purpose of investigating labor accidents with biological material involving workers and trainees occurred in 2003 and 2004 in a University Hospital of Parana. For data collection, the electronic form of the Net of Occupational Accidents Prevention - REPAT has been utilized. Out of the 586 hospital workers, there was a register of 20 (3,4%) injured workers in 2003 and 23 (3,8%) in 2004, representing an increase of 15% in the notifications from one year to the other. PMID- 16468263 TI - [Comprehension of therapeutic medication by the elderly]. AB - It is about a quantitative, descriptive and exploratory study with the objective of investigating the difficulties faced by the elderly regarding therapeutic medicaments. The sampling unit: elderly of both sexes, integrated into the area of the Health Unit totaling 45 subjects. The results have demonstrated that the most utilized medicaments were the antihypertensive ones. The reasons that interfere with adhesion to the treatment: the use of several medications concomitantly by 15 elderly; 5 out of them have not understood the medical prescription. The drawn conclusion is that the elderly consume several medications and present understanding difficulties, resulting in nonagreement with the imposed treatment and possible damages to their health. PMID- 16468264 TI - [Nurse experiencing care of patient under parenteral nutrition]. AB - It is about a research with the objective of understanding the experience of the nurse in the process of taking care being the caregiver of the patient under parenteral nutrition. We have utilized the Symbolic Interactionism and the Theory Grounded in Data for the theoretical-methodological referral. Ten nurses who have experienced this praxis in a public hospital in the city of Fortaleza, Ceara, have taken part of the study. The data collection was performed through participant observation and semi-structured interview. We have identified the phenomenon that expresses the nurse experiencing the care by showing attitudes, feelings and meanings regarding care. PMID- 16468265 TI - [Evaluation of the cardiovascular risk among climacteric women attended at a family health program]. AB - The objective of this study has been evaluating the cardiovascular risk among climacteric women attended at a Family Health Program from June to September 2003. The cardiovascular risk was assessed through the Framinghan Score. The incidence of dyslipidaemias was of 61% with hypercholesterolaemia and hypertriglyceridaemia rates of 41% and 21%, respectively. The average cardiovascular risk was of 3,5 % (+/-3,2) being higher among post-menopause women (p=0,04). These results reinforce the importance of the assistance to climacteric women in the health services in Brazil, what would contribute to the reduction of the female mortality rates. PMID- 16468266 TI - [Care concepts of caregivers: a study with hospitalized children with cancer]. AB - Qualitative research with the objective of learning the care concept of the caregiver who is a member of the multiprofessional team working with hospitalized children with cancer. Fifteen pediatric caregivers from the Hospital de Clinicas de Porto Alegre took part of it. The data collected through semi-structured interviews have been submitted to content analysis. As to the theme category, Care Conception, caregivers regard care as Action, Tie, Presence, Feelings, and Promotion of personal and spiritual development. PMID- 16468267 TI - [Teaching the group theme in graduate nursing courses in Brazil]. AB - It is about a descriptive and exploratory study with the objective of identifying Nursing Graduation Courses in Brazil that contemplate the group theme within the educational plan and its characteristics. From the data collected with Nursing Course professors in a form answered and sent back to the researchers, it has been possible to verify that teaching the group theme is little explored in the Schools, being few the common traces among the institutions regarding content, teaching hours and strategies. Due to the importance of this theme and the need of preparing the nurse for the collective work and the coordination of groups/teams, the authors recommend the insertion of this subject in the education of this professional. PMID- 16468268 TI - [The popular use of therapeutic herbs in body care]. AB - It is about a quantitative and exploratory study aimed at describing the clients' knowledge and use of phytotherapy. The theoretical referral derived from anthropology. The adopted instruments were semi-structured forms applied to 300 users in a Health Polyclinic in Niteroi, Rio de Janeiro, Brazil. RESULTS: 60,4% of the subjects use herbs and 63,4% have acquired the knowledge on medicinal plants with the family. The reasons for using herbal therapy were: sensation of improvement (32,2%) and the natural character of the herbs (33,5%). The adoption of herbs meets the subjectivity of the subject in his/her social and cultural context and remains as a complement or supplement to the official therapeutics. PMID- 16468269 TI - [Relatives of hospitalized adult patients: their participation in the nursing care process]. AB - This paper aims at unveiling the participation of relatives in the care process of hospitalized adult patients. It is about a qualitative exploratory and descriptive research with data collected through semi-structured interviews with 10 relatives of hospitalized patients in a surgical unit. The content analysis of the interviews has evidenced three categories: participation of relatives in the care; relation of the relatives with the nursing staff; and feelings of the relative regarding his/her role as caregiver. It has been noticed that most of the relatives take part in the care process, carrying out a series of activities with his/her loved one by interacting with the nursing staff. PMID- 16468270 TI - [Working with alcoholic patients: satisfaction of nurses at a general hospital]. AB - It is about a psychometric study involving 171 nurses at a general hospital in the inland of Sao Paulo state, Brazil, and aimed at verifying these professionals' satisfaction upon working with alcoholics. The results have indicated the nurses 'feelings of confusion and discomfort upon working with people who have alcohol problems. 70% of the studied subjects have indicated that they do not work well with alcoholic patients, 80%prefer not working with them and 60% face difficulties to talk about the problem with their patients. The authors suggest in site training as a strategy to better face this issue. PMID- 16468271 TI - National rural health mission. PMID- 16468272 TI - National rural health mission: from idea to action. PMID- 16468273 TI - Politics of rural health in India. AB - The setting up of the National Rural Health Mission is yet another political move by the present government of India to make yet another promise to the long suffering rural population to improve their health status. As has happened so often in the past, it is based on questionable premises. It adopts a simplistic approach to a highly complex problem. The Union Ministry of Health and Family Welfare and its advisors, either because of ignorance or otherwise, have doggedly refused to learn from the many experiences of the past, both in terms of the efforts to earlier somewhat sincere efforts to develop endogenous mechanisms to offer access to health services as well as from the devastative impact on the painstakingly built rural health services of the imposition of prefabricated, ill conceived, ill-formulated, techno-centric vertical programmes on the people of India. The also ignore some of the basic postulates of public health practice in a country like India. That did not substantiate the bases of some of their substantive contentions with scientific data obtained from health systems research reveals that they are not serious about their promise to rural population. This is yet another instance of what Romesh Thaper had called 'Baba Log playing government government'. PMID- 16468274 TI - Indian public health standards (IPHS) for community health centres. AB - The health care system in India has expanded considerably over the last few decades but the quality of the services is not up to the mark due to various reasons. Hence standards are being introduced in order to improve the quality of services. A task group under the chairmanship of Director General of Health Services, Government of India was constituted to recommend the standards to be called as Indian Public Health Standards. IPHS are a set of standards envisaged to improve the quality of health care delivery in the country under the National Rural Health Mission. PMID- 16468275 TI - National rural health mission--hope or disappointment? AB - The recently declared National Rural Health Mission has aroused significant interest, being both welcomed and closely scrutinized, since there is a long overdue and outstanding need to strengthen weak and dysfunctional public health systems in rural India. In this setting, Jan Swasthya Abhiyan (JSA) has been involved in analysing various aspects of the Mission. The concern has been that it should develop in a manner that actually strengthens public health systems in an integrated manner, and that it should empower communities to be involved in the planning and utilization of these systems in a Rights-based framework. In this article, one will draw upon and reflect on a few of the major concerns about NRHM that have emerged during the insightful discussions in JSA. PMID- 16468276 TI - Need of training for health professionals on National Rural Health Mission. AB - Training plays a key role in health manpower development and management of health care system. Since NRHM is a new concept, due weight-age has to be given to various components of the Mission. For the efficient and effective performance of medical and paramedical workers a certain level of competence in the form of knowledge, skill and attitude are essential. Various aspects of training activities to be included in training programmes have been discussed in the paper. PMID- 16468277 TI - The National Rural Health Mission (NRHM): a critical overview. AB - This paper explores the forces that led to the conceptualisation of the National Rural Health Mission (NRHM) including the role of the Common Minimum Programme and the Structural Adjustment Programme. The paper analyses the key components of the NRHM in terms of the theoretical frameworks of decentralisation, integration of programmes, primary health care, community health workers and standards. PMID- 16468278 TI - Need for dedicated focus on urban health within National Rural Health Mission. AB - National Rural Health Mission represents an important public health initiative to address essential health needs of the country's underserved population. For the Mission to achieve its goals, urban population needs to be included in its scope. Urban poor population constitutes nearly a third of India's urban population and is growing at three times the national population growth rate. Health status and access of reproductive and child health services of slum dwellers are poor and comparable to the rural population. Efforts to improve the conditions of urban poor necessitate strengthening national policy and fiscal mandate, augmenting and strengthening the urban health delivery system, coordinating among multiple stakeholders, involving private sector, strengthening municipal functioning and building community capacities. National Rural Health Mission should be broadened to National Public Health Mission. This paper discusses issues pertaining to health conditions of the urban poor, present status of services, challenges and suggests options for NRHM to bridge the large gap. PMID- 16468279 TI - National Rural Health Mission--a critical review. AB - National Rural Health Mission is strategic framework to implement the National Health Policy 2002. The scheme of Accredited Social Health Activist is an improvement over the earlier Community Health Guide Scheme. Integration of various health and family welfare programmes will result in economy and allocation of resources as per needs of the districts. Decentralised planning with the involvement of Panchayati Raj Institutions is likely to make health as people's programme. Converging water supply, sanitation, hygiene and nutrition with health planning is a logical step. The proposal to strenthen institutions of primary health care and Community Health Centres as functional Rural Hospitals alongwith introduction of Indian Public Health Standards and accountability of public health institutions to the public is likely to revolutionise the status of health care in rural India. PMID- 16468280 TI - National Rural Health Mission--hopes and fears. Concerns about targeted sterilisation, retreat of the state and privatisation. AB - The National Rural Health Mission (NRHM), launched by the present government as part of its honouring the Common Minimum Programme (CMP) commitment, had its content shaped by an active process of dialogue between many stakeholders. This article traces the contours of the discussions on three key concerns of civil society that influenced their contributions to the shaping of the National Rural Health Mission agenda. These three concerns were promotion of targeted sterilisation, a retreat of the state from its commitments to the health sector and that the NRHM agenda would lead to privatisation of public health facilities. Whereas fears on targeted sterilisation and retreat of the state may be unrealistic, there is a thrust to increased involvement of the private sector, which needs to be understood in its entirety. There is need for continued engagement byequity concerned public health professionals and health activists at all levels of implementation and not merely community monitoring to influence and shape the National Rural Health Mission in a pro-poor direction. PMID- 16468281 TI - Challenges of maternal mortality reduction and opportunities under National Rural Health Mission--a critical appraisal. AB - Maternal Mortality Ratio (MMR) continues to remain high in our country without showing any declining trend over a period of two decades. The proportions of maternal deaths contributed by direct obstetric causes have also remained more or less the same in rural areas. There is a strong need to improve coverage of antenatal care, promote institutional deliveries and provide emergency obstetric care. Delays occur in seeking care for obstetric complications and levels of 'met obstetric need' continue to be low in many parts of the country. Most of the First Referral Units (FRUs) and CHCs function at sub-optimal level in the country. National Rural Health Mission (NRHM) offers institutional mechanism and strategic options to reduce high MMR. 'Janani Suraksha Yojna', strengthening of CHCs (as per Indian Public Health standards) to offer 24 hours quality services including that of anesthetists and Accredited Social Health Activist (ASHA) are important proposals in this regard. District Health Mission can play an important role in monitoring maternal deaths occurring in hospitals or in community and thus create a social momentum to prevent and reduce maternal deaths. NRHM, however, depends largely on Panchayati Raj Institutions for effective implementation of proposed interventions and utilization of resources. In most parts of our country, State Governments have not empowered PRIs with real devolution of power. Therefore, much needs to be done locally to build the capacity of PRIs and develop state-specific guidelines in operational terms to implement interventions under NRHM for reducing maternal mortality ratio. PMID- 16468282 TI - National Rural Health Mission--"rhetoric or reality". AB - National Rural Health Mission (NRHM) launched by Government of India holds great hopes and promises to serve the deprived and underserved communities of rural areas. The backbone of the programme is Accredited Social Health Activist (ASHA), which will play major role in the implementation of the programme. The invariable existence of socio-cultural clusters in the community has always been a major challenge to the health care efforts made by the government. Though ASHA is a novel concept to melt the ice in the culture of silence among the various cluster community groups, it is important to emphasize that inter-cluster communication may still pose a problem, which ASHA may be unable to address. Considering the constraints of ASHA and success of cluster community approach in Unicef supported community based Maternal Child Health & Nutrition (MCHN) Project, it is quite reasonable to state that inclusion of community mobilisers (Bal Parivar Mitra) from within the cluster community group might well be an asset to the programme, who may actually bring about the task of spreading the spirit of NRHM. These set of functionaries may work in coordination to bring about the desired behaviour changes and decrease the social delays responsible for maternal and childhood mortality. It will also bring about the feeling of community participation and ownership. The programme is in its initial phase but has years ahead of it to bring visible changes at community level to make it a reality. PMID- 16468283 TI - National rural health mission--opportunity for Indian Public Health Association. AB - Indian Public Health Association (IPHA) welcomes the release of National Rural Health Mission (NRHM) documents. It suggests that manpower requirements of the Community Health Centre (CHC) should be rationally determined on the basis of work and patient load of the CHC. Importance should be given on availability of simple & life saving equipment, female staff when male staff is not available. Safe drinking water, an adequate sanitation and excreta disposal facility through Panchayet Raj Institution (PRI) or privatization was proposed. Accredited Social Health Activist (ASHA) has been accepted more streamlining based on the community was suggested. Capacity building or training should be CHC based for grass-root level functionaries with incentive to Medical officer (MO). IPHA proposes to extend support in capacity building, development of manual for ASHA & other categories of health professional as well as Program Implementation Plan (PIP). PMID- 16468285 TI - Clinical management of chronic disease. PMID- 16468284 TI - National Rural Health Mission (2005-2012)--Mission document. PMID- 16468286 TI - Rosacea: clinical presentation and pathophysiology. AB - Acne rosacea is one of the most common diagnoses seen in the clinical dermatologic practice. The classic presentation of rosacea, acneiform papules, and pustules on a background of telangiectasia, is often easily identified by primary care physicians, patients, or their similarly afflicted friends or family members. However, rosacea actually represents a spectrum of disease from chronic skin hypersensitivity and flushing to rhinophyma. Although the pathogenesis of rosacea remains unknown, it is important to understand its various presentations and possible etiologies prior to developing individualized treatment protocols. PMID- 16468287 TI - Oral therapy for rosacea. AB - This article will examine oral therapies utilized in the treatment of rosacea. Important topics include recognizing which types of rosacea can benefit from oral therapy and concerns regarding the emergence of bacterial resistance. PMID- 16468288 TI - Topical therapies for rosacea. AB - Therapeutic options for rosacea include topical agents, oral therapies, laser and light treatments, and surgical procedures. Topical therapies play a critical role in the treatment of patients with papulopustular rosacea and erythematotelangiectatic rosacea, and have the ability to effectively minimize certain manifestations of the disease, including papules, pustules, and erythema. The 3 primary agents for the topical treatment of rosacea are metronidazole, azelaic acid, and sodium sulfacetamide-sulfur. Each of these therapies is approved for the treatment of rosacea and has been validated by multiple studies. Additional topical therapies including benzoyl peroxide, clindamycin, retinoids, topical steroids, calcineurin inhibitors, and permethrin are not approved for the treatment of rosacea and play variable roles in the management of this condition. PMID- 16468289 TI - Treatment of rosacea with herbal ingredients. AB - Since rosacea is a chronic disease and many patients find prescription therapies unsatisfactory, they frequently turn to herbal ingredients for relief of their persistent facial redness. The most useful and frequently used herbal compounds include licorice, feverfew, green tea, oatmeal, lavender, chamomile, tea tree oil, and camphor oil. The utility of most of these herbs is based on their purported anti-inflammatory properties. Some of these herbs have proven effects, many have potential benefits, and some may aggravate rosacea. Due to the fact that many patients fail to inform their physicians about their use of herbal ingredients, dermatologists should be aware of what patients may be using and be able to advise them about the efficacy of these ingredients or the potential for adverse effects. PMID- 16468290 TI - Laser and light therapies for acne rosacea. AB - Acne rosacea is a multifactorial, somewhat mercurial disorder that can be a challenge to control with standard pharmacologic agents. Laser and light sources have been increasingly utilized, particularly for control of the generalized erythema, flushing, and telangiectasia of rosacea. This paper will review the clinical studies presented in the literature specifically treating patients with rosacea. Long-pulsed dye lasers and intense pulsed light devices can offer patients effective treatment without the purpura of short-pulsed dye lasers. Long term efficacy has not been studied but maintenance therapy may be necessary to control the vascular manifestations of this disease. PMID- 16468291 TI - Pattern of development of basal versus squamous cell carcinoma. AB - BACKGROUND: Basal cell and squamous cell carcinomas both arise in the epidermis of fair-skinned people in response to ultraviolet light, with the overall frequency of basal cell carcinoma being 4 times that of squamous cell carcinoma. Despite the similarities in the population at risk, and the presumed etiology of these tumors, it is unclear if any one individual has a proclivity to develop only one type of tumor. OBJECTIVE: The study explores whether or not there is a pattern of expression of basal versus squamous cell carcinoma among people with these cancers. METHODS: This case-control study involved patients with a total of more than 3 and fewer than 10 basal or squamous cell carcinomas. Patient age and gender, as well as number and location of diagnosed basal and squamous cell carcinomas were gathered and patterns within these values were sought. RESULTS: Patients found to have at least one basal cell carcinoma tended to produce more basal cell carcinomas and patients found to have at least one squamous cell carcinoma tended to produce more squamous cell carcinomas. CONCLUSION: The study supports the possibility that people who develop basal cell carcinoma are more likely to develop more basal cell carcinomas. Similarly, people who develop squamous cell carcinoma are more likely to develop more squamous cell carcinomas. PMID- 16468292 TI - Long-pulsed dye laser-mediated photodynamic therapy combined with topical therapy for mild to severe comedonal, inflammatory, or cystic acne. AB - BACKGROUND: Acne patients who fail to respond to conventional treatments have been treated with isotretinoin, an effective treatment coming under strict regulation due to the risk of significant side effects. Photodynamic therapy (PDT) may be a viable alternative treatment for recalcitrant acne of various types and levels of severity. OBJECTIVE: To determine the safety and efficacy of combination PDT with topical 5-aminolevulinic acid (ALA) and activation by long pulsed, pulsed dye laser (LP PDL, 595 nm) energy with topical therapy in patients with mild to severe acne. METHODS: A prospective, controlled pilot, proof-of principle study of 19 consecutive patients (aged 16-47 years, Fitzpatrick skin types I-VI) with mild to severe cystic, inflammatory, or comedonal acne of the face was conducted. All patients had failed conventional therapy, including oral antibiotics, topical treatments, hormonal therapy, laser procedures (without ALA), and/or oral isotretinoin. Fifteen patients were treated with ALA PDT and 4 patients served as controls; all were continued on topical medications. Patients undergoing PDT were initially randomized to receive either blue light or laser energy. Because recrudescence occurred in 1 patient while undergoing multiple treatments with ALA and blue light, all subsequent patients were treated with ALA and laser energy. The total number of patients treated with LP PDL-mediated ALA PDT was 14. ALA was applied for a short 45-minute incubation followed by 1 minimally overlapping pass with the LP PDL (595 nm, 7.0-7.5 J/cm2 fluence, 10-ms pulse duration, 10-mm spot size, and dynamic cooling spray of 30 ms with a 30-ms delay). Patients treated with conventional therapy (oral antibiotics, oral contraceptives, and topical medications) or laser energy without ALA PDT served as control groups. Patients were followed monthly for up to 13 months. RESULTS: Complete clearance was achieved in 100% (14 out of 14) patients in the LP PDL PDT treated group. A mean of 2.9 treatments (range 1-6; 2.0-3.7, 95% CI; n=14) was required to achieve complete clearance for a mean follow-up time of 6.4 months (range 1-13; 3.8-8.9 95% CI; n=14). The patient mean percent lesional clearance rate per treatment was 77% (64%-90%, 95% CI; n=14). Improvement in acne lesions became apparent within 1 to 2 weeks after the first treatment. Clearance in the LP PDL PDT group was superior to control groups. In the LP PDL-only control group (n=2), the patient mean percent lesional clearance rate per treatment was 32% without complete clearance after 3 to 4 treatments. In the oral antibiotics, oral contraceptives, and topicals control group (n=2), the clearance rate per treatment was 20%, the mean clearance rate per month was 4%, and complete clearance was not achieved after 6 to 10 months. In the LP PDL-mediated PDT group, treatments were well-tolerated with minimal erythema lasting 1 to 2 days. No cases of crusting, blistering, purpura, scarring, or dyspigmentation occurred. A reduction in the erythema in erythematous acne scars was observed. CONCLUSION: For teenage to adult patients with recalcitrant comedonal, inflammatory, or cystic acne of various degrees of severity, ALA PDT with activation by LP PDL appears to be a safe and effective treatment with minimal side effects. LP PDL mediated PDT may serve as an important alternative to isotretinoin. Cosmetically well-accepted, LP PDL PDT combined with topical therapy is the first PDT modality to achieve complete clearance with long-term follow-up as compared to controls. PMID- 16468293 TI - Treatment of dysplastic nevi with 5% imiquimod cream, a pilot study. AB - OBJECTIVE: To assess the clinical and histologic effects of topical imiquimod therapy on dysplastic nevi, and to determine the feasibility of using in vivo confocal microscopy (CSLM) to non-invasively monitor histological response of dysplastic nevi to imiquimod therapy. DESIGN: Single-blinded pilot study with patients not blinded as to treatment status. SETTING: Dermatology Outpatient Clinic, Memorial Sloan-Kettering Cancer Center, New York, NY. PATIENTS: The study population comprised of 10 patients with clinically dysplastic (atypical) nevi and at least 8 large nevi, (> or =5 mm) on their trunk. INTERVENTION: Sixteen weeks of imiquimod 5% cream applied to treatment lesions 3 times per week. MAIN OUTCOME MEASURE: Clinical response as gauged by comparison of baseline and week 20 1:1 standardized photographs for all study nevi and histological assessment of each patient's 4 largest study nevi at completion of therapy. RESULTS: There were no obvious clinical changes in the size and morphology of the study nevi. Subtle changes in nevus color could not be assessed due to imperfect spectral registration of images over the course of the study. Histologically, 4 of 14 treated nevi and 0 of 14 untreated nevi p=0.03 showed a significant relative reduction of junctional and intraepidermal nevocytes accompanied by papillary dermal fibroses and variable inflammation suggestive of partial regression. Non invasive CSLM imaging of study nevi demonstrated previously reported in vivo features of dysplastic nevi. but the imaging equipment and protocol utilized proved inconsistent across lesions and time. CONCLUSIONS: The histological changes seen in a subset of treated nevi suggest a possible role for the use of topical immune response modifiers for the treatment of dysplastic nevi with the intent of melanoma chemoprevention. The dose regimen of topical imiquimod utilized in this study failed to induce sufficient clinical or histological responses to warrant further study. Targeting of dysplastic nevi and intermediate endpoints for melanoma chemoprevention with more intense and/or prolonged treatment regimens with imiquimod or the use of other immune response modifiers seems promising. Technical improvements are required for the use of non-invasive CSLM imaging in lieu of invasive histology for the study of topical nevus therapies. PMID- 16468294 TI - Rebound of psoriasis during treatment with efalizumab. AB - Recently, the biologics have emerged as a new class of drugs for systemic therapy of psoriasis with efalizumab (Raptiva) being one of the most recently FDA approved agents. We report a case of a 34-year-old Caucasian male who experienced psoriasis rebound during a 7-week lapse in treatment with subcutaneous efalizumab 0.8 mg/kg/week, which he had been using for 4 weeks up to that point. The patient then restarted efalizumab, but his psoriasis continued to worsen causing him to be admitted for Goeckerman day care. He rebounded again 4 weeks following his ultimate discontinuation of efaluzimab despite intensive therapy with the Goeckerman regimen. This case is reported to highlight the recent findings that psoriasis rebound, defined as psoriasis area and severity index (PASI) of 125% or greater or morphology change, has been associated with both the use and the discontinuation of efalizumab. PMID- 16468295 TI - Bullous reaction to topical methchlorethamine in mycosis fungoides. AB - We describe a patient with mycosis fungoides who developed a bulla shortly after the start of topical methchlorethamine. Dermal-epidermal separation has been reported to occur with topical methchlorethamine on histological examination. It is plausible that the compromised dermal-epidermal junction may manifest as a clinical bulla after a secondary insult. To our knowledge, clinical bulla formation due to topical methchlorethamine has not been reported. PMID- 16468296 TI - Plantar verrucous carcinoma treated with Mohs micrographic surgery: a case report and literature review. AB - BACKGROUND: Verrucous carcinoma of the foot is a slow-growing tumor that is capable of causing vast local destruction. It is a rare tumor and is frequently misdiagnosed as a plantar wart. OBJECTIVE: To describe a patient with verrucous carcinoma of the foot treated by Mohs micrographic surgery and give a review of the literature. RESULTS: A 43-year-old male with plantar verrucous carcinoma was successfully treated with Mohs micrographic surgery. CONCLUSION: Early diagnosis of verrucous carcinoma of the foot and treatment with Mohs micrographic surgery may prevent amputation of the foot. PMID- 16468297 TI - Tinea versicolor clearance with oral isotretinoin therapy. AB - We describe the incidental clearance of preexisting tinea versicolor skin infection with the treatment of oral isotretinoin therapy for acne vulgaris. PMID- 16468298 TI - Lamotrigine-induced erythema multiforme mimicking contact dermatitis. AB - Erythema multiforme is an acute, hypersensitivity reaction of the skin often secondary to medications. Lamotrigine is a relatively new anticonvulsant medication approved for seizure and psychiatric disorders. Although the overall incidence of cutaneous reactions to lamotrigine is high, the incidence of serious eruptions such as erythema multiforme, Stevens-Johnson syndrome, and toxic epidermal necrolysis is low. Vigilant surveillance for any cutaneous eruption in patients on lamotrigine is important, particularly in the first 8 weeks, as prompt discontinuation of the medication can prevent progression. We report a case of erythema multiforme secondary to lamotrigine, which clinically resembled a contact dermatitis, and review the management of lamotrigine associated cutaneous eruptions. PMID- 16468299 TI - Exogenous ochronosis. AB - We describe a case of a 70-year-old woman who had been using a skin-lightening cream containing hydroquinone for a previous diagnosis of melasma. She presented a hyperpigmentation predominantly on her cheeks and eyebrows. The biopsy showed deposition of yellow-brown globules in the dermis. A diagnosis of exogenous ochronosis was made. An attempt of treatment using a Q-switched Nd:YAG laser has been initiated recently. PMID- 16468300 TI - New and emerging treatments for acne and photoaging. PMID- 16468301 TI - [Prospect for research on deregulation of cell proliferation and apoptosis in human gliomas]. PMID- 16468302 TI - [Review of neuropathology in the past 10 years in China]. PMID- 16468303 TI - [Retrospection of hematopathologic research of the past 50 years in China]. PMID- 16468304 TI - [Expression of apolipoprotein E in Alzheimer's disease and its significance]. AB - OBJECTIVE: To study the association between Alzheimer' s disease (AD) and apolipoprotein E (apoE) polymorphism and apoE epsilon4 allele; and to investigate the role of apoE in senile plaque formation. METHODS: During the period from 1982 to 2003, 27 portmortem cases of AD from the archival files of Department of Pathology of Beijing Hospital, diagnosed according to the consortium to establish a registry for Alzheimer's disease (CERAD) criteria, were enrolled into this study. Among the 27 cases studied, there were 23 cases of definite AD and 4 cases of probable AD. Postmortem brain tissues from 67 neurologically unremarkable deceased were used as age-matched controls. Immunohistochemical study for beta amyloid (Abeta) and Tau protein, as well as immunohistochemical study for Abeta/apoE, were performed in all AD cases using streptavidin-peroxidase (SP) and double immunostaining ( SP/ABC) methods, respectively. Senile plaques and neurofibrillary tangles in the 23 cases of definite AD were further quantified. The apoE genotypes in all cases were analyzed by polymerase chain reaction and restriction fragment length polymorphism technologies. RESULTS: Immunohistochemical study for Abeta distinguished 4 different types of senile plaques: diffuse non-neuritic plaques, diffuse neuritic plaques, dense-core neuritic plaques and dense-core non-neuritic plaques. Double immunohistochemistry for Abeta/apoE showed that some senile plaques were positive for both Abeta and apoE. The expression rates for Abeta and apoE in these 4 different types of senile plaques were 4. 28%, 84. 71%, 8.50% and 2.51%, respectively. The positivity rate for Abeta/apoE in diffuse neuritic plaques were significantly higher than those in other 3 types (P < 0.01). The frequency of occurrence of apoE epsilon4 allele in AD was significantly higher than that in the control group (P < 0.01). The numbers of senile plaques and neurofibrillary tangles in AD cases with apoE epsilon4 allele were also significantly higher than those in AD cases without apoE epsilon4 allele (P < 0.01). CONCLUSIONS: ApoE polymorphism is associated with AD. The presence of apoE epsilon4 allele carries a higher risk for the development of AD. ApoE may also play an important role in the transformation of diffuse non-neuritic plaques to diffuse neuritic plaques. PMID- 16468305 TI - [A clinicopathologic study of dysembryoplstic neuroepithelial tumor]. AB - OBJECTIVE To study the clinicopathologic features, radiologic findings, treatment modalities and prognosis of dysembryoplastic neuroepithelial tumor (DNT). METHODS: The clinical features, histopathologic findings, immunohistochemistry and electron microscopy of 18 cases of DNT were analyzed. Results Among the 18 cases studied, 14 were males and 4 females. The age of these patients ranged from 3 to 46 (mean age = 22. 8 years). Partial seizure was the main presenting symptom in all patients. The history of epilepsy could be as long as 17 years. On magnetic resonance imaging (MRI) study, the tumor was hypodense on T1 and hyperdense on T2. There was neither edema nor mass effect. All but 2 cases were supratentorial and intracortical in location. Ten cases were treated by complete surgical excision and the remaining 8 tumors were partially excised. In the 14 patients with follow-up data available, 13 survived for 1.4 to 11 years after the operation (with more than 10 years survival observed in 2 patients). The average survival period was 5.5 years. None of the cases showed tumor recurrence after operation. Histologically, all tumors demonstrated a multinodular architecture and were intracortical in location, sometimes with extension into the white matter. The characteristic "glioneuronal constituent" was an essential feature for making the diagnosis of DNT. The tumor was formed by an admixture of oligodendrocyte-like cells, mature neurons and astrocytes, with obvious microcystic changes. These neurons were often dispersed singly in the mucoid matrix. In most cases, the foci of cortical dysplasia were found in adjacent areas. Immunohistochemical study demonstrated positivity for synaptophysin, neurofilament and S-100 protein in the neurons and some oligodendrocyte-like cells. The staining of glial fibrillary acidic protein in the oligodendrocyte like cells was negative. Electron microscopy showed early neuronal, astrocytic and oligodendroglial differentiation of the oligodendrocyte-like cells. CONCLUSIONS: DNT is a benign tumor (corresponding to WHO grade I) that can be cured by surgical excision, despite sometimes incomplete tumor removal. A correct diagnosis of this entity requires thorough understanding of the clinical, radiologic, histologic and immunohistochemical features. PMID- 16468306 TI - [Clinicopathologic and radiologic features of dysembryoplastic neuroepithelial tumors]. AB - OBJECTIVE: To study the clinicopathologic features and radiologic findings of dysembryoplastic neuroepithelial tumor (DNT). METHODS: The clinical presentations, radiologic findings, histologic features and immunophenotype of 9 cases of DNT were analyzed. RESULTS: The age of patients ranged from 12 to 51 years (mean age = 32 years). Most presented with partial seizures, sometimes accompanied by transient aphasia, agraphia and decreased visual acuity. One case was asymptomatic and discovered incidentally during a routine check-up. All patients had no neurological deficit found on physical examination. All tumors were located in the supratentorial cerebral cortex. There was no peritumoral edema or space-occupying effect on radiologic examination. The tumors involved either frontal lobe (number = 4), temporal lobe (number = 4), frontoparietal lobe (number = 1) . Two cases showed cystic changes. Two histologic variants of DNT were recognized: simple (number = 3) and complex (number = 6). Simple variant was composed mainly of the glioneuronal element, accompanied by surrounding oligodendrocyte-like cells, and the complex variant contained a low-grade glioma component, in addition to the glioneuronal element and sometimes foci of cortical dysplasia. CONCLUSIONS: DNT is a benign tumor with excellent prognosis after surgical excision. Local recurrence is rare. Complex variant of DNT needs to be distinguished from other types of low-grade glioma. PMID- 16468307 TI - [Study of HOXA5 gene expression in breast carcinoma]. AB - OBJECTIVE: To study mRNA and protein expression of HOXA5 gene in breast carcinoma, to correlate the expression of HOXA5 gene with clinicopathologic parameters and to explore the possible role of HOXA5 gene in carcinogenesis, progression and metastasis of breast carcinoma. METHODS: TaqMan real-time reverse transcriptase-polymerase chain reaction (RT-PCR) was applied on 60 cases of primary breast carcinoma and 24 cases of benign mammary lesions in order to detect mRNA expression of HOXA5 gene. Immunohistochemical study using polyclonal antibody against HOXA5 was also performed. Statistical analysis was carried out to analyze the correlation between HOXA5 gene expression and various clinical parameters in these breast cancer patients. RESULTS: (1) The relative expression level of HOXA5 mRNA ranged from 0.73 to 193.07 (average = 20.85) in primary breast carcinoma, in contrast to 5.42 to 81.91 (average = 30.94) in benign mammary lesions. Compared with benign mammary lesions, a significant reduction in expression of HOXA5 mRNA was noted in primary breast carcinoma (P < 0.01). (2) There was a decreased or completely diminished HOXA5 protein expression in breast carcinoma. (3) HOXA5 mRNA expression was significantly lower in lymph node positive cases, when compared with that in lymph node-negative cases (P < 0.05). A significant difference of HOXA5 protein expression was also observed in both groups (P < 0.01). Immunohistochemical staining of HOXA5 was either negative or weakly positive in lymph node-positive cases. On the other hand, moderately or strongly positive HOXA5 staining was noted in lymph node-negative cases. (4) Neither mRNA nor protein expression of HOXA5 gene correlated with clinicopathologic parameters such as age of patients, size of tumor, clinical stage, pathologic subtype or histologic grade (P > 0.05). CONCLUSIONS: Disordered expression of HOXA5 gene may play a role in the carcinogenesis of breast cancer. Reduced expression of HOXA5 gene may be related to the metastatic potential of breast carcinoma cells. PMID- 16468308 TI - [Pathologic feature and diagnosis of pulmonary alveolar proteinosis]. AB - OBJECTIVE: To describe the pathologic features and diagnostic algorithm of pulmonary alveolar proteinosis (PAP). METHODS: Thirty-nine biopsy and postmortem cases of PAP were studied by light microscopy and histochemical staining using periodic acid-Schiff (with digestion) (PAS-D), mucicarmine (with digestion) (mucicarmine-D) and alcian blue. RESULTS: Histologically, the affected lung tissue displayed the following characteristic features: (1) alveoli and some of the small bronchioles were filled with eosinophilic and fine granular proteinaceous material with needle-like clefts; (2) proteinaceous material was seen admixed with various numbers of degenerated and sometimes exfoliated pneumocytes; (3) pneumocytes were hyperplastic; (4) alveolar capillaries and alveolar septa had become hyperemic, but pulmonary interstitial inflammation was not obvious; (5) no significant inflammation was identified in the bronchial wall; (6) compensating emphysema was noted in the surrounding lung parenchyma. Fragments of eosinophilic, finely granular proteinaceous material with needle like clefts were also found in the bronchoalveolar lavage fluid under light microscopy. The proteinaceous material was stained red by PAS-D. The staining for mucicarmine-D was negative, while alcian blue staining was either weakly positive (faint blue staining) or negative. Pathologic examination of lung biopsies and bronchoalveolar lavage fluid thus remaines the gold standard for diagnosis of PAP. CONCLUSIONS: Identification of homogeneous, eosinophilic, finely granular and PAS-D-positive proteinaceous material with needle-like clefts in alveolar spaces or bronchoalveolar lavage fluid is of diagnostic importance in PAP. Bronchoalveolar lavage, being a relatively safe and non-invasive procedure, can be a useful adjunct in arriving at the final conclusion. PMID- 16468309 TI - [VEGF-induced tubulogenesis of endothelial cells from human brain malignant glioma in the three dimentional model]. AB - OBJECTIVE: To compare the tubulogenesis capability of malignant glioma-derived microvessel endothelial cells (GDMEC) from human brain with that of ECV304 cells in a three dimentional model and to explore the significance of GDMEC in the study on angiogenesis. METHODS: The GDMEC were isolated from malignant gliomas of human brain and purified by selective binding to the monoclonal antibody against CD105 bound to the magnetic MACS MicroBeads. GDMEC and endothelial-like cell line ECV304 were compared with their capabilities of formatting tubule-like structure (TLS) in the three dimentional collagen matrix, with or without inducement by various concentration of vascular endothelial growth factor (VEGF). RESULTS: The obtained GDMEC had a high purification (98%) and could be successfully cultured in vitro. GDMECs formed more TLS than ECV304 cells of the same number and at the same time points. VEGF could induce rapid formation of TLS in a dose-dependent manner, however, ECV304 cells were less response to VEGF stimulation. CONCLUSIONS: GDMEC could maintain their endothelial characteristics and potential capability of angiogenesis. They were more response to VEGF than ECV304, therefore, more suitable for in vitro studies on tumor angiogenesis. PMID- 16468310 TI - [Poorly-immunogenic tumor is capable of inducing proliferation of CD4 (+) CD25 (+) regulatory T-lymphocytes in vitro]. AB - OBJECTIVE: To investigate the distribution of regulatory T-lymphocytes in the splenocytes cocultured with syngeneic low-immunogenic tumor cells, as compared with that of highly-immunogenic tumor cells, to investigate the mechanism underlining tumor evasion. METHODS: Three different immunogenic tumor cells were cocultured with syngeneic splenocytes individually to mimic cancer immunity in vitro. The proliferation response of splenocytes was measured by thymidine incorporation. The distribution of TR cells, CD4(+) IFN-gamma (+) T cells and CD4(+) IL-10(+) T cells were analyzed by flow cytometry. The secretion of IFN gamma and IL-10 in supernatants was measured by ELISA assay. RESULTS: The stimulation Index of splenocytes cocultured with syngeneic highly-immunogenic H22 or FBL3 was much higher than that of poorly immunogenic melanoma D5. In each group, stimulation Index of splenocytes cocultured with allogeneic tumor cells was higher than that of the corresponding tumor immunity model. In addition, compared with those of highly-immunogenic tumors, there were more TR, CD4(+)IL 10(+) and less CD4 (+)IFN-gamma(+) T cells in the splenocytes, and higher IL-10 and lower IFN-gamma levels in the supernatant of the splenocytes stimulated with low-immunogenic D5 cells. CONCLUSION: Poorly-immunogenic tumor cells can induce the proliferation of TR cells, which may play an important role in tumor evasion. PMID- 16468311 TI - [Targeted inhibition of vascular endothelial growth factor (VEGF) expression in human osteosarcoma cell line by antisense VEGF165 cDNA promoted by hypoxia reaction element]. AB - OBJECTIVE: Utilizing the hypoxia inducible factor 1/hypoxia reaction element (HIF 1/ HRE) gene regulation system to construct antisense vascular endothelial growth factor (VEGF165) cDNA eukaryotic expression vector promoted by HRE, and investigate its targeted inhibiting VEGF expression of osteosarcoma cells in hypoxia environment. METHODS: Eukaryotic expression plasmid with HRE promoter was constructed containing luciferase reporter gene and antisense VEGF165 cDNA by using PCR and recombinant DNA techniques. The recombinant vectors were transfected into osteosarcoma cells with lipofectin method. Hypoxia-inducible reporter gene expression was determined by liquid scintillation analyzer and the expression of VEGF protein was detected by ELISA method. RESULTS: The eukaryotic expression plasmid containing antisense VEGF165 and luciferase promoted by HRE was constructed successfully. After being transferred into MG63 cells, luciferase expression was increased 3.5 x 10(2) times and VEGF protein expression decreased 45% under hypoxia condition. CONCLUSION: Antisense VEGF165 cDNA expression, efficiently realized by HRE promoter under hypoxia condition, provides an experimental basis for targeted antiangiogenesis of tumors. PMID- 16468313 TI - [Exploring the significance of NET-1 gene in hepatocellular carcinoma]. PMID- 16468312 TI - [Hepatitis B virus and deletion of its COOH-terminal forty amino acids: proliferative impact on hepatoma cell line SMMC-7721]. AB - OBJECTIVE: To explore the biological impact of 40 amino acid deletion at the C terminal of hepatitis B virus X on the proliferation of hepatoma cells. METHODS: Cells of SMMC-7721 hepatoma cell line were transfected with HBx and its derivative HBx3'-40, harboring the 40 amino acid deletion at the distal C terminal region. Cell growth curve, colony formation in soft agar plate and tumorigenesis assay in nude mice were used to observe the alterations induced by the transfection of HBx and HBx3'-40. The expression level of PCNA in tumor cells was also investigated. RESULTS: The growth rates of the cells transfected with HBx and HBx3'-40 were markedly increased as compared with that of the control group. The colony formation rates were enhanced in the cells transfected with HBx(48.7 +/- 8.1) and HBx3'-40 (82.8+/-6.0), comparing with the control (26.9 +/- 3.5) %. In the tumorigenic assay, the size and weight of tumors were significantly increased in the cells transfected with HBx (0.412 +/- 0.212, 0.395 +/- 0.159) % and HBx3'-40 (1.476 +/- 0.232, 0.987 +/- 0.279) %, as compared with the control group (0.051 +/- 0.024, 0.033 +/-0.004) %. The expression level of PCNA in tumors was increased in both HBx (59.00 +/- 2.58) % and HBx3'-40 (69.25 +/- 3.77) % transfected cells, comparing with the control (37.67 +/- 2.52) %. Overall, the cells transfected with HBx3'-40 demonstrated the highest proliferative capacity. CONCLUSION: The deletion of 40 amino acids in the C terminal of HBx is correlated with an enhanced proliferation of hepatoma cells and may play an important role in the malignant transformation of the liver. PMID- 16468314 TI - [Dynamic distribution of L. interrogans in guinea pigs and pathologic changes in experimental leptospirosis]. PMID- 16468315 TI - [The roles of important molecules of Wnt signaling pathway in non-small-cell lung cancer]. PMID- 16468316 TI - [The inhibiting effect of polygoni multiflori total glycosides (PMTG) on the atherosclerotic lesion formation in experimental atherosclerosis]. PMID- 16468317 TI - [The expression of BP1 mRNA and its clinical significance]. PMID- 16468318 TI - [Dysplastic gangliocytoma of cerebellum (Lhermitte-Duclos disease)]. PMID- 16468319 TI - [Multiple splenosis of the mediastinum: a case report]. PMID- 16468320 TI - [Infantile rhabdomyofibrosarcoma]. PMID- 16468321 TI - [Anaplastic diffuse large B-cell lymphoma involving liver: case report]. PMID- 16468322 TI - [Non-specific steroid cell tumor of the ovary with myelolipoma]. PMID- 16468323 TI - [Signet ring cell carcinoma arising from mature cystic teratoma of the ovary]. PMID- 16468324 TI - [A case of intestinal neuronal dysplasia (type B)]. PMID- 16468325 TI - [Multiplex fluorescence in situ hybridization in detecting complex chromosomal aberrations in myelodysplastic syndromes]. AB - OBJECTIVE: To explore the value of multiplex fluorescence in situ hybridization (M-FISH) technique in the detection of the complex chromosomal aberrations (CCAs) in myelodysplastic syndromes (MDS). METHODS: M-FISH was used in ten MDS patients with R-banding CCAs to refine the complex chromosomal rearrangements, the constitute of marker chromosomes, and to identify the cryptic translocations. RESULTS: Thirty-seven kinds of structural rearrangements were detected by M-FISH including insertion, deletion, translocation and derivative chromosomes, among which 34 kinds were unbalanced rearrangements, and 3 were balanced rearrangements including t(6;22) (q21; q12), t(9; 19) (q13; p13) and t(3;5) (?; ?). Seven abnormalities in the present paper were first reported in the literature. In addition, chromosome 17 aberrations (7/10) and -5/5q - (7/10) were the two most frequent abnormalities. CONCLUSIONS: M-FISH could refine CCAs in MDS patients, find or correct the missed or misidentified abnormalities analysed by conventional cytogenetics. PMID- 16468326 TI - [Study on DNA methylation status of WT1 gene in its promoter region in hematologic malignancy cell lines]. AB - OBJECTIVE: To study the DNA methylation status of WT1 gene promoter region in hematologic malignancy cell lines and its correlation with WT1 gene expression. METHODS: 1. RT-PCR and methylation-specific PCR were performed for detecting WT1 gene expression and DNA methylation status in its promoter region in 8226, HL-60, Jurkat, K562, KG-1, NB4, Raji, SHI-1, U266 and U937 cell lines. 2. Treatment of U937 cells with 5-aza-CdR, a demethylation inducing agent and the changes in WT1 gene expression level and its promoter region methylation status were determined. RESULTS: 1. HL-60, K562, KG-1, NB4 and SHI-1 cells showed higher levels while 8226, Jurkat, Raji, U266 and U937 cells showed extremely low levels of WT1 expression. DNA hypermethylation in WT1 gene promoter region was identified in 8226, Jurkat, Raji, U266 and U937 cells. 2. The WT1 gene expression in U937 was enhanced after treatment with 5-aza-CdR accompanied with the decrease of methylated and the increase of unmethylated levels in its promoter region. CONCLUSION: Modulation of the DNA methylation status in WT1 promoter region is one of the epigenetic mechanisms for regulating its expression. PMID- 16468327 TI - [Identification of two novel mutations in ADAMTS13 gene in a patient with hereditary thrombotic thrombocytopenic purpura]. AB - OBJECTIVE: To investigate the gene mutations of ADAMTS13 in a highly suspected hereditary thrombocytopenic purpura (TTP) patient, and then make a progressive diagnosis and adjust the plan of therapy. METHODS: ADAMTS13 activity and inhibitor were determined by residual-collagen binding assay during several episodes. Genomic DNA extracted from the proband's peripheral blood was used for amplification of 29 exons and exon/intron boundaries of ADAMTS13 by PCR. The PCR products were screened by direct sequencing and the gene alterations were further confirmed by direct sequencing in her family members. RESULT: The activity of the proband's ADAMTS13 was significantly reduced while no inhibitor was found. Two novel missense mutations were found in the TSPI repeated motif domain of ADAMTS13. In both mutations, thymine substituted for cytidine, resulting in the substitution of leucine for serine in nt 2708, exon 21 (codon S903L), and tryptophan for arginine in nt 3283, exon 25(codon R1095W). These two mutations were revealed as each heterozygote in the proband's parents. CONCLUSION: The deficiency of ADAMTS13 caused by two homozygote missense mutations might be responsible for episode of this TTP patient. PMID- 16468328 TI - [Study on platelet-associated tissue factor and its significance]. AB - OBJECTIVE: To explore whether normal platelet contains tissue factor (TF), and the significance of platelet-associated TF (PATF). METHODS: Platelets were isolated by Sepharose 2B gel column. ELISA was used to detect the TF content in the lysates of washed platelets. Procoagulant activity of PATF was measured by one stage clotting time assay. The mRNA of TF was detected by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: A certain amount of TF antigen (16.37 +/- 6.39) ng/L was detected in the washed-platelet lysates. Upon activation by collagen, platelets released TF and caused a marked increase in TF level in plasma (P <0.05). Resting platelets had no TF procoagulant activity, while procoagulant activity of platelets activated by collagen increased significantly, which could be blocked by TF McAb and poor VII plasma. TF mRNA could not be detected in washed platelets. TF content in platelets from patients with coronary heart disease was significantly higher than that from normal controls (P < 0.05). Resting platelets from the patients showed a higher procoagulant activity, which could be inhibited by TF McAb. CONCLUSION: Platelets contain TF and the latter released by activated platelet was functionally active. Platelet itself might not synthesize TF. Protein content and procoagulant activity of PATF in patients with coronary heart disease were higher than that in controls. All these indicate that platelet may be involved in coagulation and thrombosis by releasing TF. PMID- 16468329 TI - [Expression of coagulation factor IX in human CD34 + hematopoietic stem cells by adeno-associated virus 2]. AB - OBJECTIVE: To investigate the expression of human coagulation factor IX (hFIX) gene in human umbilical cord blood (CB) CD34+ cells which was transfected with recombinant adeno-associated virus 2 (rAAV-2). METHOD: The CD34+ cells were transfected with rAAV-2/hFIX and cultured for 21 days for inducing differentiation into myeloid, erythroid and megakaryocytes, respectively. The expression of hFIX was studied at mRNA, protein and biological activity levels. The cytotoxicity of rAAV-2 to CD34+ cells was evaluated by cell proliferation, cell vitality, CD antigen expressions and CFU yields. RESULTS: The hFIX mRNA was expressed in the cultured cells which was verified by RT-PCR and DNA sequencing. An elevated level of hFIX expression and biological activities were detected with a maximum amount of 14.10 ng/10(6) cell x 24 h. During the period of 21 day culture, the cell vitality, cell proliferation, CD antigen expression and CFU yields between the transfected and un-transfected groups had no difference(P > 0.05). CONCLUSION: The human CB CD34+ cells are able to produce functional hFIX after transduction by rAAV-2/hFIX. The cell proliferation and differentiation capacities of the host CD34+ cells were not affected by rAAV-2. PMID- 16468330 TI - [Regulation of tissue factor expression in brain microvascular endothelial cells by PLA nanoparticles coating NF-kappaB decoy oligonucleotides]. AB - OBJECTIVE: To investigate a new strategy of polylactic acid (PLA) nanoparticles delivery system coating nuclear factor-kappaB (NF-kappaB) decoy oligonucleotides (ODNs) for inhibiting TF expression in cultured brain microvascular endothelial cells(BMECs). METHODS: PLA nanoparticles coating FITC-labeled NF-kappaB decoy ODNs were formulated by nano-deposition method and the characteristics of nanoparticles were detected. BMECs were isolated and cultured in vitro. The cellular uptake and intracellular localization of nanoparticles in BMECs was detected by flow cytometry and confocal microscopy. Changes in the expressions of TF and nuclear protein P65 were examined by RT-PCR and Western blot in NF-kappaB decoy ODNs transfected BMECs by LPS stimulation. RESULTS: The decoy-nanoparticles obtained were uniform spherical particles with an effective diameter of 162.1 nm and a polydispersity index of 0.118. NF-kappaB decoy ODNs encapsulated in nanoparticles could be released in a controlled manner in phosphate-buffered saline for up to 28 days. It was observed that the cellular uptake of nanoparticles were increased with the time of incubation and the concentration of nanoparticles in the medium. Nanoparticles localized mainly in the BMECs cytoplasm. LPS-induced upregulation of TF transcription was inhibited by NF kappaB decoy ODNs transfection but not by missense ODNs transfection. Furthermore, changes in the transcription level of TF were paralleled by a reduction of capacity of P65 in nuclear extract of NF-kappaB decoy ODNs transfected cells. CONCLUSIONS: These findings offer a potential therapeutic strategy in the control of TF expression in BMECs in vitro. PMID- 16468332 TI - [The influence of different WT1 gene isoforms expression pattern on the differentiation of leukemia cell line NB4]. AB - OBJECTIVE: To study the potential effects of exogenous WT1 gene isoform on the differentiation of leukemia cell line NB4 and its possible molecular mechanisms. METHODS: The recombinant eukaryotic expression vector (pCB6 + /WTA) containing full-length human WT1 isoform (WTA: -17AA/ -KTS) cDNA and the blank pCB6 + vector were transfected into leukemia cell line NB4 by electroporation. The WTA mRNA and protein in cells were detected by RT-PCR and Western blot. Cell morphology, NBT reduction and CD11b antigen expression in NB4 cells were assayed to evaluate cell differentiation. Expression of PML/RARalpha, p21 and c-myc genes was determined by semi-quantitative RT-PCR after transfection. RESULTS: Compared with NB4/WTA cells, NB4 and NB4/CMV (NB4 cells transfected with pCB6 + vector) cells had higher morphological differentiation rates and higher CD11b expression levels after exposure to ATRA for 48 hours. The percentage of NBT reduction in NB4/WTA cells was lower than that in control groups. The difference in NBT reduction rate between NB4/WTA and control cells was gradually increased after treated with ATRA for three days. The expression levels of PML/RARalpha, p21 and c-myc genes in NB4/WTA cells were notably increased. CONCLUSION: Overexpression of exogenous WTA gene could partially inhibit the differentiation of NB4 cells by up-regulating the expression of PML/RARalpha, p21 and c-myc genes. PMID- 16468331 TI - [The effects of pravastatin on platelet-derived nitric oxide system in rabbits]. AB - OBJECTIVE: To observe the effects of pravastatin on platelet-derived nitric oxide system in hypercholesterolemia (HC) and atherosclerosis (AS) in rabbits, and the relationship between these changes and atherosclerosis courses. METHODS: Thirty male New Zealand white rabbits were randomly divided into three groups, 12 in group A, 12 in group B, and 6 in group C. All of them were fed daily with cholesterol-rich food during the first 12 weeks. In addition, in group A, pravastatin (10 mg) was orally administered daily. At the end of the 12th week, 6 in group A and B were killed randomly and their aortas were removed and the pathologic changes were observed. In the following 12 weeks, food enriched with cholesterol was substituted with normal food in all three groups. Pravastatin treatment was continued or started in the remaining members of group A and group B, but not in group C. At the end 24th week, all rabbits were killed and their aortas were examined for the fatty-streaks or atherosclerotic plaques. The expressions of endothelial NOS (eNOS) mRNA and inducible NOS (iNOS ) mRNA, NOS activity, NO production and the level of the serum lipids were measured at 0, 6th, 12th, 18th and 24th week. RESULTS: The expression levels of platelet-derived NOS mRNA, eNOS mRNA ratio in group A had no difference at above time points, while in group B were reduced significantly at 6th week and 12th week compared with at 0 week (P <0.01), and increased at 18th week and 24th week compared with 12th week (P <0.05). The expression levels of eNOS mRNA in group C were reduced at 6th, 12th and 18th, 24th week compared with 0 week (P <0.05 and P <0.01, respectively), and were reduced in groups B and C compared with group A at 6th ,12th week (P < 0.05) and increased in group A and B compared with group C at 18th, 24th week (P <0.01). The expression levels of iNOS/mRNA among the three groups had no difference. Pathologic finding of the arteries: AS was not found in group A from the 12th to 24th week. While in group B, there were a lot of fatty streaks on the entire intima of all large arteries at the 12th week. There were also fatty-streaks in the ascending aorta, but were improved at the 24th week. In group C, there were marked plaques in the entire aorta at the 24th week. CONCLUSIONS: The expressions of platelet-derived eNOS mRNA, NOS activity, NO production are decreased in HC or AS rabbits. Pravastatin can up-regulate expressions of platelet-derived eNOS mRNA, NOS activity, leading to preventing or improving the pathological courses of AS. PMID- 16468333 TI - [Pure red cell aplasia following major ABO-incompatible allogeneic hematopoietic stem cell transplantation]. AB - OBJECTIVES: To study clinical characteristics and outcome of pure red cell aplasia (PRCA) following major ABO-incompatible allogeneic hematopoietic stem cell transplantation (allo-HSCT). METHODS: Variables including sex, age, stem cell source, granulocyte engraftment time, blood transfusion and isoagglutinin type against donor RBC were analyzed to identify risk factors for the development of PRCA. RESULTS: Twelve of 100 patients received major ABO-incompatible allo HSCT developed PRCA, with out any effect on incidence of aGVHD and CMV infection. ABO blood groups of recipient/donor pairs of these twelve PRCA patients were O/A in nine, B/A in one and O/B in two. Patients with anti-A isoagglutinins against donor RBC developed PRCA more frequently than those with anti-B (10/49 vs 2/49). Median duration to the recovery of erythropoiesis tended to be longer in patients with PRCA (PRCA vs non-PRCA, 203.5 vs. 76 days, P < 0.05 ). Median durations to the disappearance of incompatible isoagglutinins tended to be longer in patients with PRCA (PRCA vs. non-PRCA, 150.5 vs. 60 days,P <0.05) and in those with anti-A isoagglutinins (anti-A vs anti-B, 90 vs 55 days, P < 0.05). CONCLUSION: ABO blood group of O/A in recipient/donor pair was the only high risk factor for PRCA after major ABO-incompatible allo-HSCT. PMID- 16468334 TI - [Effects of curcumin on the acetylation of histone H3, P53 and the proliferation of NB4 cells]. AB - OBJECTIVE: To investigate the effects of curcumin on the acetylation of histone H3, P53 and the proliferation of NB4 cells. METHODS: The total protein was extracted from NB4 cells treated without or with different concentrations of curcumin ( 50, 25, 12.5, 6.25 and 3.125 micromol/L) for different time (0, 4, 8, 12, 24 h). Western blot analysis was performed to determine the levels of acetylated histone H3, P53 and acetylated P53. MTT assay was performed to examine the growth inhibition effect of curcumin on NB4 cells. RESULTS: Curcumin could inhibit the proliferation of NB4 cells in a time- and dose- dependent manner, with the IC50 at 24 h and 36 h of 40 micromol/L and 25 micromol/L, respectively. The levels of histone H3 acetylation, P53 expression and P53 acetylation were increased obviously. CONCLUSION: Curcumin functions as a deacetylase inhibitor,which could increase the level of acetylated histone H3, enhance the expression and activity of tumor suppressor P53, and inhibit the proliferation of NB4 cells. PMID- 16468335 TI - [Inhibition effect of antisense Bmi-1 on Jurkat cells]. AB - OBJECTIVES: To investigate whether antisense Bmi-1 plasmid could inhibit the proliferation of Jurkat cells. METHODS: The antisense plasmid was constructed by PCR amplification of a 171 bp segment spanning Bmi-1 start codon and zinc finger structure and the PCR product was subsequently inserted reversely to plasmid pLNCX2. The final construct was confirmed through restriction enzyme digestion. G418 was added into the medium after the plasmid was successfully introduced into Jurkat cells by using lipofectin-mediated DNA transfection. The proliferation of Jurkat cells were determined by MTT and colony formation assays. Cell cycle was determined by flow cytometry. The p16 expression of Jurkat cells was studied by immunofluorescent histochemistry. RESULTS: The growth rate of antisense Bmi-1 transfected Jurkat cells was significantly lower than that of the controls, and the colony forming capacity of the transfected cells decreased significantly (P < 0.01), the colony numbers being (90.7 +/- 9.07)/10(3) cells, (83.3 +/- 6.11)/10(3) cells and (56.0 +/- 5.56)/10(3) cells for control cells, empty plasmid transfected Jurkat cells and antisense Bmi-1 transfected Jurkat cells, respectively. The percentage of G, phase cells was increased and the p16 expression of antisense Bmi-1 transfected cells was significantly upregulated than that of control cells. CONCLUSION: Antisense Bmi-1 can inhibit the growth and upregulate the expression of p16 of Jurkat cells in vitro. PMID- 16468336 TI - [Genomic DNA sequences and functional expression, purification of BmalphaTX14 neurotoxin from scorpion Buthus martensii Karsch]. AB - Based on the full-length cDNA of BmalphaTX14 from Chinese scorpion Buthus martensii Karsch (BmK), gene of the mature peptide of BmalphaTX14 was cloned into the yeast expression vector pPIC9K. After transforming, screening and inducing, tricine-SDS-PAGE and Western blot proved that rBmalphaTX14 protein was expressed in the medium for up to 84 hours, getting nearly 120 mg/L. Recombinant BmalphaTX14 was purified rapidly and efficiently through Ni-NTA-agarose, polyethylene glycol precipitation and gel filtration chromatography. The purified rBmalphaTX14 proved to have the anti-insect activity by toxicity assay. Meanwhile, genomic gene of BmalphaTX14 was cloned and sequenced by PCR method, sequence analysis of this gene showed that BmalphaTX14 had an intron of 408 base pairs located at the signal peptide encoding region, which was similar with the characteristic of other alpha-type sodium ion-channel toxin. Considering both the genomic organization and the peptide function, BmaTX14 proved to be a membership belonging to alpha-type sodium ion-channel toxin. PMID- 16468337 TI - [Construction and immunogenicity of recombinant pseudorabies virus expressing the modified GP5m protein of porcine reproduction and respiratory syndrome virus]. AB - Pseudorabies virus (PRV), an alpha-herpesvirus, has been used as a vector for live-viral animal vaccines. The recombinant PRV TK- / gE- / GP5+, which expressing GP5 of PRRSV, is developed based on the PRV genetic-depleting vaccine virus strain, TK- / gE- /LacZ+. However, this strain stimulated poorly the vaccinated animals to produce neutralizing antibodies against PRRSV. In order to develop a booster specific immunized response of the PRV recombinant, the ORF5 gene of PRRSV TK- / gE- / LacZ+ was substituted by a modified ORF5 gene, ORF5m. The resultant recombinant PRV, TK- /gE- / GP5m+, was verified by PCR, Southern blotting and Western blotting. TK- / gE- / GP5m+ and TK- / gE- / GP5+ expressed GP5 proteins were inoculated into balb/c mice to evaluate their immunogenicity. The results demonstrated that the amount of neutralization antibodies and cell immunity responses induced by TK- / gE- /GP5m+ against PRRSV were higher than that of TK- / gE- / GP5+. This study indicated that the new recombinant PRV expressing the modified GP5m protein is a candidate for the development of bivalent genetic engineering vaccines against PRRSV and PRV. PMID- 16468338 TI - [Salmonella choleraesuis C500 delivering DNA immunization against classical swine fever virus]. AB - Classical Swine Fever Virus (CSFV) E2 protein eukaryotic expression plasmid pVAXE2 was constructed. The plasmid pVAXE2 was transformed into Salmonella choleraesuis C500 (S. C500) attenuated vaccine strain by electroporation to generate Salmonella choleraesuis engineering strain S. C500/pVAXE2. The characterization of S. C500/pVAXE2 in morphology, growth, biochemistry and serology indicated that it retained the same properties as its original strain S. C500 with exception of kanamycin resistance originated from the plasmid pVAXE2. The plasmid stable in the bacteria after 15 passages. Kunming mice and rabbits were vaccinated three times at two weeks interval with S. C500/pVAXE2 in oral and intramuscular routes at the dosage of 1 x 10(8) CFU for mice and 2 x 10(9) CFU for rabbits each time. The specific antibody response against CSFV and Salmonella choleraesuis was detected by ELISA. Two weeks after the third boost the immunized rabbits were challenged with 20 ID50 of hog cholera lapinized virus (HCLV), followed by a virulent strain of Salmonella choleraesuis two week later than HCLV challenge. The results showed that all immunized mice and rabbits produced significant antibodies against CSFV and Salmonella choleraesuis, and the immunized rabbits demonstrated the effective protection against the challenge of HCLV and virulent Salmonella choleraesuis. These results indicated the potential of developing multiplex swine DNA vaccine by using this bacteria as the vector. PMID- 16468339 TI - [Heteroexpression of Rhizopus arrhizus delta6-fatty acid desaturase gene in Pichia pastoris]. AB - Delta6-fatty acid desaturase is a membrane-bound enzyme, which is rate-limiting for the biosynthesis of polyunsaturated fatty acids. A cDNA sequence putatively encoding a delta6-fatty acid desaturase was isolated from Rhizopus arrhizus NK300037 using RT-PCR and RACE methods in our previous work. Sequence and function analysis indicated that this sequence was a novel delta6-fatty acid desaturase gene which had an open reading frame of 1377bp coding 458 amino acids of 52kD. The methylotrophic yeast Pichia pastoris, has been developed into a highly successful system for the production of a variety of heterologous proteins during the past 20 years. In this work, the Rhizopus arrhizus delta6-fatty acid desaturase gene (RAD6) was subcloned into expression vector pPIC3.5K to generate a recombinant plasmid pPICRAD6, which was subsequently transformed into Pichia pastoris strain GS115 for heterologous expression by electroporation method. Total fatty acids were extracted from the induced cells and methylated. The resultant fatty acid methyl esters (FAME) were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Fatty acids analysis showed that the coding product introduced a new double bond at delta6 position of appropriate fatty acid substrates including C16:1, C17:1, C18:1, linoleic acid and alpha-linolenic acid without chain length specificity of fatty acids. Furthermore, modification of sequence flanking AUG codon of this delta6-fatty acid desaturase gene increased the expression of target gene in P. pastoris. All of these results suggest that P. pastoris is an optimal expression system of delta6-fatty acid desaturase gene. PMID- 16468340 TI - [Expression of endo-beta-mannanase gene from Trichoderma reesei in Pichia pastoris]. AB - Complete mannanase gene with two introns was cloned from Trichoderrna reesei by PCR. The two introns were then removed by overlap extension PCR. The gene encoding the mature mannanase protein was inserted into the expression vector pPIC9K, downstream of a alpha-factor signal peptide sequence. The resultant recombinant vector was named pM242. After linearized with Sac I , pM242 was transformed to Pichia pastoris GS115 by electroporation. After screening, the recombinant strain Gpmf25 that expresses the secretory protein at high level was obtained. The activity of the recombinant mannanase reached 12.5 IU/mL. Optimum pH and temperature for the recombinant enzyme were 5.0 and 80 degrees C, respectively. The enzyme was stable at pH 5.0-6.0 and maintained over 50% of original activity after incubation at 70 degrees C for 30 min. PMID- 16468341 TI - [Construction of multifunctional genetically engineered pesticides-degrading bacteria by homologous recombination]. AB - Construction of multifunctional pesticides-degrading genetically engineered microorganisms (GEMs) is increasing important in the bioremediation of various pesticides contaminants in environment. However, construction of genetically stable GEMs without any exogenous antibiotic resistance is thought to be one of the bottlenecks in GEMs construction. In this article, homologous recombination vectors with the recipient's 16S rDNA as homologous recombination directing sequence (HRDS) and sacB gene as double crossover recombinants positive selective marker were firstly constructed. The methyl parathion hydroalse gene (mpd) was inserted into the 16S rDNA site of the carbofuran degrading strain Sphingomonas sp. CDS-1 by homologous recombination single crossover in the level of about 3.7 x 10-(7) - 6.8 x 10(-7). Multifunctional pesticides-degrading GEMs with one or two mpd genes inserted into the chromosome without any antibiotic marker were successfully constructed. The homologous recombination events were confirmed by PCR and southern blot methods. The obtained GEMs were genetically stable and could degrade methyl parathion and carbofuran simultaneously. The insertion of mpd gene into rrn site did not have any significant effect on recipient' s physiological and original degrading characteristics. The methyl parathion hydrolase (MPH) was expressed at a relatively high level in the recombinants and the recombinant MPH specific activity in cell lysate was higher than that of original bacterium (DLL-1) in every growth phase tested. The highest recombinant MPH specific activity was 6.22 mu/tg. In this article, we describe a first attempt to use rRNA-encoding regions of Sphingomonas strains as target site for expression of exogenous MPH, and constructed multifunctional pesticides degrading GEMs, which are genetically stable and promising for developing bioremediation strategies for the decontamination of pesticides polluted soils. PMID- 16468342 TI - [Cloning and expressing of cellulase gene (cbh2) from thermophilic fungi Chaetomium thermophilum CT2]. AB - Chaetomium thermophilum CT2 can produce extracellular cellulase with industrial value. We designed two degenerate primers to amplify catalytic domain sequence of cellobiohydrolase II ( CBH II). Full length of cDNA was obtained by rapid amplification of cDNA ends technologies. DNA sequencing revealed that cbh2 has an open reading frame of 1428bp, which encodes a putative polypeptide of 476 amino acids. The deduced amino acid sequence shows that the predicted molecular mass is 53 kD and the cbh2 consists of a fungal-type carbohydrate binding domain (CBD) separated from a catalytic domain by a linker region rich in proline/serine/threonine. PCR product consisting of the entire CBH II coding region without its signal sequences was cloned into the yeast secretive plasmid pPIC9K, which was then transformed into Pichia pastoris GS115. Highly efficient production of the cellobiohydrolase II was achieved in P. pastoris under the control of the AOX1 promoter, and the expressing level was 1.2 mg/mL by small scale culturing. The recombinant cellobiohydrolase II was purified by using ammonium sulfate fraction, DEAE-Sepharose Fast flow chromatography. A molecular mass of the purified enzyme is 67 kD determined by SDS-PAGE and this is similar to the native cellobiohydrolase II purified from C. thermophilum CT2. The recombinant enzyme exhibited optimum catalytic activity at pH 4.0 and 50 degrees C respectively. It was thermostable at 50 degrees C and retained 50% of its original activity after 30 min at 70 d degrees C . The high level of fully active recombinant cellobiohydrolase II got from P. pastoris makes this expression system attractive for fermentor and industrial applications. PMID- 16468343 TI - [Construction and screening of phage display single chain antibody library against Bursaphelenchus xylophilus cellulase]. AB - A phage display single-chain variable fragment (scFv) library against Bursaphelenchus xylophilus cellulase (BXC) was constructed and used to screen the specific antibodies binding to BXC. The total RNA was extracted from fresh spleens of BALB/C mice immunized with BXC. Gene fragments encoding VH and VL were amplified by RT-PCR and assembled into a single chain by overlapping PCR with a linker DNA encoding the peptide (Gly4Ser)3. The recombinant fragments were cloned into the phagemids (pCANTABSE) and electroporated into E. coli TG1. The recombinant phagemids were rescued by reinfection of helper phage M13K07. The repertoire of the phage display antibody was about 5 x 10(4). The specific antibodies against BXC were obtained after five rounds of affinity selection. The positive phage clone was used to infect E. coli HB2151. SDS-PAGE and western blot analysis showed that the soluble scFv antibodies expressed bound specifically to BXC. The studies laid foundation for quarantine and pathological study of Bursaphelenchus xylophilu. PMID- 16468344 TI - [Isolation and characterization of a DRE-binding transcription factor from Yinxin poplar (Populus alba x P. alba var. pyramidalis)]. AB - Dehydration-Responsive Element Binding ( DREB) transcription factors, specifically binding with dehydration reponsive element (DRE), activate a variety of stress-responsive genes in plants under abiotic stresses (dehydration, high salt and low temperature). Using PCR and homologous EST search, we isolated a DREB-like gene from Yinxin poplar (Populus alba x P. alba var. pyramidalis) named PaDREB2. Yeast One-hybrid experiment demonstrated that PaDREB2 protein could function as a DREB transcription factor activating target gene expression by specifically binding to DRE cis-element. To study the expression pattern of PaDREB2, RT-PCR was carried out. And the results showed that PaDREB2 is induced by low temperature, drought and high salt. PMID- 16468345 TI - [High level expression of PNGase F in Escherichia coli and its bioactivities]. AB - In order to obtain active recombinant PNGase F in Escherichia coli, a prokaryotic expression vector pET28a/PNGase F was constructed. Amplification of PNGase F was obtained using PCR technique employing suitable primers designed according to the PNGase F gene sequence from Flavobacterium nmeningosepticum. The expression of PNGase F gene in LB medium or M9 medium led to the formation of inclusion body and soluble protein, respectively. The refolding of the denatured inclusion body was successful by gradual dilution. Further purification of the refolded protein and soluble crude extract from M9 medium with Ni2+ -NTA argarose resulted a 90% purified PNGase F. The purified protein catalyzed the complete and intact cleavage of N-linked oligosaccharides from various glycoproteins. The efficiency of this cleavage was affected by the substrate status in the reaction system. In summary, we have developed an enzyme production system where PNGase F was over expressed in recombinant E. coli. This system can provide more than 15 mg/L purified active PNGase F. This purified active PNGase F can be used as tools in analyzing the oligosaccharide structure. PMID- 16468346 TI - [Establishment and optimization of in vitro regeneration system for Plantago major L]. AB - Plantago major is not only used as medicinal herb but also an important model plant of ecology. Little work has been reported on the tissue culture of P. major. A reproducible system for direct shoot morphogenesis and callus induction of Plantago major L. 'Giant Turkish' was described. Using seed as explants, the adventitious buds were obtained 4 to 5 weeks following incubation on MS medium supplemented with 0.2 mg/L IAA and 1.0 mg/L TDZ. The frequency of adventitious buds was as high as 100%. The average number of buds per explant was 14.6. Random amplified polymorphic DNA analysis on 9 regenerants indicated that somaclonal variation occurred at DNA level. Using leaves as explants, calli were easily induced on MS medium supplemented with 1.0 mg/L NAA 3 weeks following inoculation. The frequency of callus induction can be as high as 98%. On MS medium containing 4.0 mg/L 6-BA, 25% of calli differentiated and the mean number of buds per piece of callus was 2.8. The buds developed roots on 11/2 MS medium and formed plantlets, 90% of which survived when transplanted to greenhouse. PMID- 16468347 TI - [Characteristics of cell cycle and metabolism in microencapsulated K562 cell culture]. AB - Human K562 leukemia cells were cultured under free and microencapsulated condition, respectively. The cell cycles in the two kinds of cultures were investigated by flow cytometry. Moreover, mathematical model was established to simulate the cell viability and metabolized characteristic in different cultures. It was found that the cell percent in S phase was higher and the cell viability was better when cultured in microcapsule than that in free culture. The results showed that the model successfully described the substrate consumption and product formation in microencapsulated culture as well as in suspension culture. Based on the model, it was indicated that not only there was a higher proliferation and metabolic activity but also the time of the high activity could keep longer in microencapsulated culture. The parameters of the model showed that there was no significant difference between the two kinds of cultures when the influence of the glucose on the cell viability was concerned (kA(free) approximately = kA (APA)) but lactate had a obvious suppression effect on cell viability in free culture, and neglectable suppression in microencapsulated culture (kL(free) > kL(APA)). PMID- 16468348 TI - [Purification of a ginsenoside-Rb1 hydrolase from Helix snailase]. AB - Through a combination of twice DEAE chromatography by NaCl stepwise and gradient elution with gel filtration chromatography, a kind of ginsenoside-Rb1 hydrolase from crude Helix snailase was separated. The hydrolase was purified to apparent homogeneity on SDS-PAGE. It was estimated that the purified hydrolase was consisted of four identical subunits with a molecular mass of 110-115 kD by SDS PAGE and gel filtration chromatography. The Km and Vmax values for ginsenoside Rb1 were calculated to be 0.790 mmol/L and 10.192 micromol/(min x mg) of protein respectively. The ginsenoside-Rb1 hydrolase could only hydrolyze the glycosidic bond at the C20 position of ginsenoside-Rb1 into ginsenoside-Rd. PMID- 16468349 TI - [Comparative proteomic analysis of cerebral cortex between middle cerebral artery occlusion rats and normal controls]. AB - In order to provide a complete picture of pathogenesis in cerebral ischemia, cerebral cortex in MCAO rats were analysed for alteration in their proteomes. Comparative proteome analysis was used to compare signal corresponding to individual cerebral cortex proteins on a two-dimensional gel between MCAO rats and the normal control (NC) group. After sample preparation, two-dimensional electroghoresis separated proteins were stained with Commassie Brilliant Blue. The image data were analyzed on a Dell computer using Image Master v 3.01 software. In cerebral cortex, 30 proteins were differentially expressed in MCAO rats compared with NC. There were 11 spots significantly increased, 15 spots significantly decreased and Adenylate kinase isoenzyme 1 was detected only in NC group, biliverdin reductase B, small inducible cytokine A4 [Precursor] only in MCAO group. Peroxiredoxin 2 divided into two points in MCAO6h group. In the end, this approach may lay a foundation for the further investigation of pathogenic mechanisms in cerebral ischmic injury. PMID- 16468350 TI - [Genetically modified industrial brewing yeast with high-glutathione and low diacetyl production]. AB - Recombinant plasmid pICG was constructed by replacing the internal fragment of a acetohydroxyacid synthase (AHAS) gene (ILV2) with a copy of gamma glutamylcysteine synthetase gene (GSH1) and copper chelatin gene (CUP1) from the industrial brewing yeast strain YSF31. YSF31 was transformed with plasmid pICG linearized by Kpn I and Pst I. A recombinant strain with high-glutathione and low diacetyl production was selected. The results of fermentation in 100-L bioreactor showed that the lagering time of beer produced for recombinant strain T2 was shortened by 3 days and the shelf life of the beer was prolonged about 50%. It may be more acceptable for the commercial application, as it does not contain foreign DNA. PMID- 16468351 TI - [Polyethylene glycol-accompanied ion-exchange chromatography to purify recombinant hepatitis B virus surface antigen]. AB - The dissociation of virus-like particles of Hepatitis B surface antigen (HBsAg) during the adsorption-desorption on the solid-phase of chromatography is a main challenge for its purification. Herein, poly (ethylene glycol) (PEG) was applied as an additive during the purification of HBsAg from recombinant Chinese hamster ovary (CHO) cell culture to improve the HBsAg recovery and protect its structural assembly. The presence of 1% of PEG10000 in the mobile phase of ion-exchange chromatography (IEC) of DEAE-Sepharose FF column could increase the recovery of HBsAg from about 55% to 80%, with a similar purification (-fold) (about 12) compared with the absence of PEG. Importantly, glycosylated protein forms of HBsAg were reserved well by PEG-accompanied chromatography. Furthermore, size exclusion chromatography-multiangle laser light scattering (SEC-MALLS) analysis was performed on line to monitor the aggregates, particle size and molecular weight distribution of HBsAg. The results demonstrated that the HBsAg particle size and assembly are more homogenous after adding PEG in the mobile phase of IEC than no PEG added in the mobile phase. PMID- 16468352 TI - [Screening of a triazophos-degrading strain and pathway of its degradation]. AB - A Triazophos-degrading strain, Klebsiella sp. E6, was identified by soil enrichment technology from the soil sampled from the vicinity of a factory manufacturing Triazophos (TAP). The nutrient requirement of the strain is simple. It can use TAP as the sole sources of carbon, nitrogen and phosphorus. Comparison of the degradation rates revealed that the strain degraded TAP most effectively when TAP was used as a sole nitrogen source. No inhibition effect occurred when TAP concentration was high as 1000 mg/L in the case of TAP was used as the sole nitrogen source. Analysis of the intermediates of TAP metabolism indicated that TAP is firstly hydrolyzed into 1-phenyl-3-hydroxy-1,2,4-triazole and O,O-diethyl phosphorothioic acid. 1-phenyl-3-hydroxy-1,2,4-triazole was further mineralized into inorganic compounds. A degradation pathway of TAP was proposed. The experiment results demonstrated that the strain has potential in biodegradation of TAP pollutions. PMID- 16468353 TI - [A study on the pattern recognition of thermophilic and mesophilic proteins]. AB - Pattern recognition of thermophilic and mesophilic proteins were studied through principle component analysis, partial least-square regression and BP neural network. The results showed that the fitting accuracy of the three methods was 92%, 95% and 98%, respectively. And the forecasting accuracy was 60%, 72.5% and 72.5%, respectively. The best forecasting accuracy for thermophilic proteins was 75%, and for mesophilic proteins was 85%. A mathematical model was established and the biological meaning of it was expatiated on, a new method to discriminate the thermophilic and mesophilic proteins based on their sequences was established here. PMID- 16468354 TI - [Study on rhG-CSF modified with polyethylene glycol]. AB - Monomethoxy Polyethylene Glycol(mPEG20000) was activated by N-hydroxysuccinimede and analyzed by infrared spectrum and hydrolysis kinetics. In order to propose the optimized reaction conditions of mono-PEGylated rhG-CSF, orthogonal design of the experiment was investigated. Ion exchange chromatography was used to separate and purify PEGylated rhG-CSF from unPEGylated rhG-CSF. The purity of mono PEGylated rhG-CSF was analyzed by high performance liquid chromatography (HPLC) to be 97%. PMID- 16468355 TI - [Isolation and functional analysis of tobacco MARs]. AB - Two new MAR segments (M14 and M17) were cloned from tobacco genome. Both of the sequences contained several typical consensus sequences of MARs, which were different from the original MAR sequence, such as 90%AT-box, A-box, T-box, the base unpairing regions (BUR), autonomously replicating sequences (ARS), the consensus sequence for topoisomerase II, MAR recognition sequence (MRS), origin of replication (ORI), curved DNA motifs and ATATTT et al. To investigate the effects of these two sequences on gene expression in transgenic plants, 3 plant expression vectors were constructed with uidA gene coding beta-glucuronidase (GUS) which were flanked on one side and on both sides by the MARs we obtained. These plant expression vectors with one or two MARs were transformed into tobaccos via Agrobacterium-mediated transformation method, with the plant expression vector pCAMBIA2301 without MAR and wild type tobacco as controls. GUS histochemical staining results showed that the uidA gene expressed stably in transgenic tobaccos. Quantitative detection of GUS activity showed that the MARs could increase GUS expression levels in vivo in contrast to the controls, wherever they were flanked on one side or both sides of uidA gene. The vector ligated with MARs in the same direction on both sides of uidA could increase the GUS expression level much better than both vectors which just ligated with single MARs on one side. The former one increased the average GUS activity for 3.14 folds, but 1.56 and 2.43 folds for the latter two vectors with single MARs respectively contrasting to the pCAMBIA2301 control. But the expression differences among individual transformants were still obvious. Therefore, it was concluded that the DNA sequences we obtained in this experiment were two novel MARs and could enhance gene expression in vivo. In the meanwhile, although the numbers of the MARs typical motifs in M14 were more than in M17, especially the 90% AT box which had been considered to be the highest correlative motif with binding strength in vitro, the enhancement of gene expression was lower yet, which implied no correlation between improvement of gene expression and binding strength between MARs and nuclear matrix in vitro. PMID- 16468356 TI - [The expression of porcine circovirus type 2 ORF2 gene in insect cells and its character]. AB - To produce the recombinant baculovirus transfer plasmid pFast-ORF2, the ORF2 gene of Porcine Circovirus type 2 (PCV2) was subcloned into baculovirus transfer vector (pFastBac(TM1) ) using Bac-to-Bac baculovirus expression system. E. coli DH10Bac (Gibco BRL) containing baculovirus shutter vector (bacmid) and helper vector was transformed with recombinant plasmid pFast-ORF2. Within E. coli DH10Bac, the ORF2 gene was transposed into the bacmid. The colonies of E. coli containing recombinant bacmid (Bac. ORF2) were collected by blue/white selection. The Bac. ORF2 was transfected into sf9 cells to yield AcNPV carrying the PCV2 ORF2 gene, referred to as Ac. ORF2. Expression of the ORF2 gene of PCV2 was confirmed by indirect immunofluorescent assay (IIFA), SDS-PAGE and Western blotting. The expressed ORF2 gene product had a molecular mass of 28kD and could be recognized by the positive serum of PCV2. The results indicated the ORF2 gene was properly expressed in sf9 cell. It was noteworthy that many self-assembled virus-like particles (VLPs) were found in purified and phosphotungstic acid (PTA) stained PCV2 ORF2 protein by electron microscope. The particles were of similar morphology to the PCV2 virion and some self-assembled virus-like particles had darkly stained centers that made them appear to be empty capsids. Both PCV2 particles and self-assembled particles were approximately 17 nm in diameter. PMID- 16468357 TI - [CDNA microarray on differentially expressed genes of adipose tissue in two breeds chicken]. AB - cDNA microarray containing 9024 cDNAs was used to construct gene expression profile in order to screen differentially expressed genes of adipose tissue between broiler and Bai' er and investigate the molecular mechanism related with body fatness traits between the two breeds. Sixty seven differentially expressed genes, being involved in fat metabolism, energy metabolism, cytoskeleton, transcription and splicing factor, protein synthesis and degradation, were screened out. Furthermore, some genes that had no annotation in GenBank were screened out, they were presumed to be unknown new genes. The roles that they may play in chicken fat metabolism need clarify later. PMID- 16468358 TI - [Improving thermostability of Aspergillus niger phytase by elongation mutation]. AB - The phytase gene phyA(m) from Aspergillus niger N25 was recombined into E. coli expression vector pET-30b(+). Recombined at expression vectors pET30b-FphyA(m) was served as a template to amplify phytase gene, and the PCR product named elongation mutation gene phyA(e) was expanded with a 13 amino acid sequence from pET-30b-FphyA(m) vector at C-terminal of phytase gene phyA(m). Furthermore, phyA(e) gene was recombined into expression vector pPIC9k and expressed in Pichia pastoris. The comparison experiment of mutant phytase PP-NP0 with wild-type phytase PP-NP(m)-8 showed that: the optimum temperature of PP-NPe was increased by 3 degrees C, and its thermostability was increased by 21% when it was exposed to 10 min at 75 degrees C. Its effective reaction pH range with catalysis efficiency above 70% was pH 4.6 - pH 6.6, and wider 0.4 pH value than that of wild-type phytase. PMID- 16468359 TI - [Biological synthesis of L-ascorbyl palmitate]. AB - Biological synthesis of L-Ascorbyl Palmitate in organic system were studied in this text. The contradiction between conversion of vitamin C and concentration of L-Ascorbyl Palmitate were resolved. High conversion of vitamin C and concentration of L-Ascorbyl Palmitate were obtained by Novo435. A series of solvents(log P from -0.24 to 3.5 )were investigated for the reaction,and acetone was found to be the most suitable from the standpoint of the enzyme activity and solubility of L-ascorbic. And the equilibrium of the reaction was affected by the addition of the molecular sieves and temperature. Reaction carried out at 60 degrees C and with 20% 0.4nm molecular sieves is good for the enzyme to keep its activity and for making the equilibrium go to the product. With 1.094 g palmitic acid, 0.107 g vitamin C and 0.020 g Novo435, rotate rate of 200 r/min, the conversion of ascorbic reached 80% and the concentration of L-ascorbyl palmitate is 20 g/L after 48 h. Furthermore, reaction batch of Novo435 and substrates recycle were observed, the result indicated that Novo435 may used 4-5 times continuously with high conversion. And 6-O-unsaturated acyl L-ascorbates were synthesized through Novo435 condensation of ascorbic acid and various unsaturated fatty acids with high conversion in this text. PMID- 16468361 TI - [Determination of the catalytic structures of methyl parathion hydrolase]. AB - Methyl parathion hydrolase (MPH) is a novel member of organophosphorus hydrolase. In this study, mpd gene was expressed in Escherichia coli DH5alpha with its native promoter. MPH was purified to homogeneity. Results show that metal chelating compounds cannot inhabit the enzyme activity. Inductively Coupled Plasma-Atomic Emission Spectrometry analysis showed that MPH is a zinc-containing enzyme, the Zinc to enzyme molar ratio is near 2:1. In order to investigate critical residues related to enzymatic activity of MPH, chemical modification reagents EDC, DEPC, butanedione and pyridoxal were tested. Experiment results suggested that aspartate, glutamate, arginine and lysine are not important for enzyme activity. But DEPC, which can modify histidine residue, inactivate the enzyme activity greatly, and the inactivation rate is 9.6 h(-1). This result reflects that histidine residues are essential for enzyme activity. All these results provide basic data for MPH structure and molecular evolution research. PMID- 16468360 TI - [Purification and renaturation of recombinant human Cu, Zn-SOD by metal-chelating affinity chromatography]. AB - Overexpression of recombinant Human Cu, Zn-Superoxide Dismutase (rhCu, Zn-SOD) in E. coli results in the form of insoluble inclusion body. Purity of rhSOD inclusion body was over 80% by isolation and purification. After preliminary renaturation by conventional dilution or dialysis, enzyme preparations was respectively purified by using Copper Metals-Chelating Affinity Chromatography (Copper-MCAC). RhSOD specific activity purified by MCAC (from the sample renatured partly by dialysis) was 2.2 times as much as that by dialysis and protein recovery was 64%. RhSOD specific activity purified by MCAC (from the sample renatured partly by dilution) was 5.3 times as much as that by dilution and protein recovery was 25%. The two rhSOD preparations purified by MCAC had specific activities about 5000 u/mg and activity recoveries were all over 130% of the enzyme activities in the samples renatured partly by dilution or dialysis. The above-mentioned results indicated that Copper-MCAC resulted in a purification and further renaturation of target protein. SDS-PAGE showed that the target protein rhSOD (19 kD) was purified homogeneously and NBT activity identification proved that the purified and renatured rhSOD had very strong SOD activity. In conclusion, Copper Metals-Chelaing Affinity Chromatography appears to be a simple, rapid and efficient procedure for purifying and further renaturing rhCu, Zn-SOD by dilution or dialysis. The method provided a new idea for purifying and renaturing recombinant proteins expressed in the form of inclusion body in E. coli. PMID- 16468362 TI - [Effects of sucrose and light on the growth and production of secondary metabolites in Pueraria phaseoloides hairy roots]. AB - Effects of sucrose concentrations and light on the growth and production of total isoflavones and puerarin in Pueraria phaseoloides hairy roots cultured onto solid MS media supplemented with 1%, 3%, 5%, 7% and 9% sucrose, respectively, were investigated. The results showed that among the sucrose concentrations tested, 3% sucrose in the medium enhanced the growth and stimulated accumulation of total isoflavones and puerarin in P. phaseoloides hairy roots, After cultured for 20 days, the biomass of hairy roots reached 0.48 g (DW dry weight)/flask and its contents of total isoflavones and puerarin were 25.44 mg/g (DW) and 11.64 mg/g (DW), respectively. In comparison with 3% sucrose, the dry weight proliferation of hairy roots cultured with 5% sucrose was increased by 7.0%, while cultured with 1%, 7% and 9% sucrose, the dry weight proliferation of hairy roots was decreased by 62.4%, 42.8% and 65.3%, their total isoflavones content was decreased by 57.4%, 13% and 33.4% and their puerarin content was decreased by 47.9%, 15.8% and 35.1%; but their content of total soluble sugars was increased 0.52, 1.45 and 1.54 times, respectively. Compared with hairy roots in blue light and white light, the biomass of hairy roots cultured in the dark for 30 days was 0.83 g (DW)/flask and was increased by 37.1% and 23.3%, respectively. The content of total isoflavones in hairy roots cultured in white light was as much as 1.15 times and 1.19 times that in blue light and in the dark, respectively. It was also observed that hairy roots cultured in blue light and white light partly became light green and that blue light could inhibit accumulation of puerarin in hairy roots and the puerarin content in hairy root cultured in white light and in the dark were 1.61 times and 1.52 times that in blue light, respectively. PMID- 16468364 TI - [Systems biology and traditional Chinese medicine (TCM) modernizatin]. AB - Systems biology is a new field in biology that aims at system-level understanding of biological systems. Based on the remarkable progress from molecular biology, systems biology focuses its research on investigating the biological systems to form their entirety. Considering the trend of TCM modernization and the coherence on the point of holism between systems biology and TCM, a comprehensive way for the research of TCM modernization was referred, which involves a new concept, componentomics. PMID- 16468363 TI - [Filter-based template preparation for rapid and sensitive PCR detection of Staphylococcus aureus in meat]. AB - In the present study, an assay using Polymerase Chain Reaction (PCR) was developed for the detection of Staphylococcus aureus in meat. Based on flotation and solvent extraction technology, FTA filter was used to extract S. aureus DNA from artifically contaminated meat. Primers targeting the thermostable nuclease gene (nuc) were used to amplify a 279 bp DNA fragment which was confirmed by DNA sequencing. The detection limit of PCR was 10 cfu x g(-1) meat of S. aureus. This novel FTA-PCR assay allows for detection of Staphylococcus aureus in meat in <6 h, which is 12-24 h less than that of conventional PCR with enrichment method. Seventy-two samples were analyzed and the detection rate using the standard cultivation method was 70.8%, detection time was 5d. The detection rate of PCR amplification using filters was 73.6%, detection time was 6h. The detection rate of Baird-Parker R.P.F method was 69.4%, detection timewas 18 h. The detection rate of petrfilm RSA method was 61.1%, detection time was 18 h. Thus PCR amplification using filters provides a faster and more sensitive method of S. aureus detection than the standard cultivation method. At the same time,it provides an universal process for preparing DNA template. PMID- 16468365 TI - [Study on identification of wild and cultirated Radix Scutellariae in different growing years]. AB - OBJECTIVE: To identify Radix Scutellariae (Huangqin) of different growth years, to distinguish whether it's wild or cultivated and to provide useful information for the quality control of Huangqin crude drug. METHOD: By using morphological and histological methods, we studied 87 individuals of 45 specimens from 12 habitats of 5 provinces of China, which grew wild or were cultivated in different growing years. Moreover, 22 commercial samples of Huangqin from 7 provinces were also investigated. RESULT: The identification was performed base on morphological and histological characteristics, such as, the shape, color, cork, remaining stems, decayed central xylem, and vessels arrangement, xylem cork ring, growth rings, etc. CONCLUSION: We established an identification method for distinguishing Huangqin wild or cultivated in different growing years. Furthermore, the structure of annual rings in the transection of Radix Scutellariae has been discovered for the first time. PMID- 16468366 TI - [Study on circumscription of medicinal licorice species based on molecular biology]. AB - OBJECTIVE: Elucidate the medicinal licorice species circumscription in order to develop licorice resources in China and select new cultivars. METHOD: PCR amplification, DNA sequencing and cladistic analysis. RESULT: Acquire the ITS sequences and phylogenetic tree of 8 licorice species. CONCLUSION: According to results Glycyrrhiza eurycarpa and G. glabra var. glandulosa should be combined into G. and G. glabra respectively and G. egladulosa is an independent species. PMID- 16468367 TI - [Studies on fingerprints of ant nest of Macrotermes annadalei from different areas]. AB - OBJECTIVE: To study comparatively the fingerprint of zhuang medicinal material ant nest of Macrotermes annadalei from different areas in Guangxi. METHOD: The fingerprint of total amino acid of 12 lots of ant nest of M. annadalei from different areas was determined by high speed amino acid analyzer. Separation was perform on anylytical column (4.6 mm x 40 mm), column temperature at 57 degrees C. The flow rate of buffering solution was 0.4 mL x min(-1), and the flow rate of ninhydrin was 0.3 mL x min(-1). RESULT: 18 characteristic peaks in the fingerprints were identified in 12 lots of samples, the chromatographic overlap rate was 89.4%-100.0%, and the total relative peak area of 7 lots of samples was over 52% in eight main peaks. CONCLUSION: The components of amino acid of ant nest of M. armadalei from different areas in Guangxi are very similar. The qualities of majority samples are good as well. The fingerprints can provide the useful information for the quality evaluation and the identification of ant nest of M. annadalei. PMID- 16468368 TI - [Investigation on the formula of Fuyankang dispersed tablets]. AB - OBJECTIVE: To investigate the preparation technique and optimal formulation of Fuyankang dispersed tablets. METHOD: The formula of Fuyankang dispersed tablets were optimized in terms of disintegrating time by studying single factor and orthogonal design test. RESULT: The products formulated with the optimum techniques met the quality specification of dispersed tablets. The dissolubility of the optimized dispersed tables was obviously faster than that of common tablets. CONCLUSION: This prescription and technology of Fuyankang dispersed tablets are reasonable and effective. PMID- 16468369 TI - [Determination of C21 steroidal glycosides in Marsdenia tenacissima by colorimetric method]. AB - OBJECTIVE: To establish a quantitative method to deter mine C21 steroidal glycosides in Marsdenia tenacissima. METHOD: Methanol was used as the extraction solvent and the samples were purified by macroporous resin ADS-7. A mixture of sulfuric acid-methanol (4:1)was used as color-producing reagent. The absorbance was measured at 325 nm. RESULT: There is a good linearity (r = 0.999 9, n = 6) within the range of 10.6-148.4 microg. The average recoveries of tenacissoside-H at different concentrations were 95.8% to 97.1% with RSD less than 2.4% (n = 5). CONCLUSION: This method is reliable as a quantitative analytical method for the quality assessment of M. tenacissima. PMID- 16468370 TI - [Determination of molecular weight and content of Ginkgo biloba exocarp polysaccharides by HPLC]. AB - OBJECTIVE: To determine the molecular weight and content of Ginkgo biloba exocarp polysaccharides. METHOD: The analysis was carried on a PL aquagel-OH MIXED (7.5 mm x 300 mm, 8 microm) chromatography column eluted with water as mobile phase at 1.0 mL x min(-1) of flow rate, the column temperature was 25 degrees C and the eluate was detected by RID. RESULT: The average molecular weight of Ginkgo bilobaexocarp polysaccharides was 11 062.5 with RSD = 0.78% (n = 6); the content was 81.9% with RSD = 2.5% (n = 6), the standard curves of dextran (MW 12 000) were linear in the range of 1-20 microg, r = 0.999 9. The average recovery is 97.9%, RSD was 2.5%. CONCLUSION: This method was found to be sensitive and accurate for the measurement of Ginkgo biloba exocarp polysaccharides. PMID- 16468371 TI - [Chemical constituents from root of Prismatomeris tetrandra]. AB - OBJECTIVE: To study the chemical constituents from root of Prismatomeris tetrandra. METHOD: Compounds were isolated by chromatographic techniques on silica gel column. Their structures were elucidated by chemical and spectral methods. RESULT: Six compounds were identified as 1-hydroxy-2-methylanthraquinone (I), 2-hydoxy-3-methoxyanthraquinone (II), 1, 3-dihydroxy-2-methoxyanthraquione (II), rubiadin (IV), rubiadin-1-methyl ether (V) and beta-sitosterol (V). CONCLUSION: I, II and III were isolated from the plant for the first time. PMID- 16468373 TI - [Studies on the alkaloids from herb of Corydalis adunca]. AB - OBJECTIVE: To study the alkaloid constituents of Corydalis adunca. METHOD: The constituents were isolated on silica gel column and their structures were elucidated by IR, NMR, MS data. RESULT: Four alkaloid compounds were isolated from alcohol extract of the herb of C. adunca, and identified as tetrahydropalmatine, 1, 2, 3, 4-tetrahydro-6-methoxy-1-(4'-methoxyphenyl) methyl 7-isoquinolinol, coptisine and palmatine. CONCLUSION: Four compounds were isolated from C. adunca for the first time. PMID- 16468372 TI - [Studies on the chemical constituents in root of Coleus forskohlii]. AB - OBJECTIVE: To study the chemical constituents in root of Coleusforskohlii. METHOD: The chemical constituents were isolated by column chromatography. The structures were elucidated on the basis of IR, MS, 1H-NMR, 13C-NMR and 2D-NMR experiments. RESULT: Six compounds were obtained and the structures were identified as 14-deoxycoleon U (1), demethylcryptojaponol (2), alpha-amyrin (3), betulic acid (4), alpha-cedrol (5) and beta-sitosterol (6). CONCLUSION: Compounds 1 and 2 were isolated from the genus Coleus for the first time. Compounds 3 and 4 were isolated from C. forskohlii for the first time. PMID- 16468374 TI - [Effects of Astragalus membranaceus and Panax notoginseng on the transformation of bone marrow stem cells and proliferation of EPC in vitro]. AB - OBJECTIVE: To investigate the effect and the possible mechanism underlying the promotional effect of Astragalus membranaceus and Panax notoginseng on the transformation of bone narrow stem cells and proliferation of EPC. METHOD: The marrow blood was collected in the patients with ischemia of lower limbs and BM MNCs were separated and proliferated under different conditions. A. morphologic observation was performed and the ratio of CD34+ cells was measured. RESULT: The shuttle shaped cells lined up as bunches with several round cells scattered. The ratio of CD34+ cells was significantly increased in groups treated with medium (P < 0.01) and lower (P < 0.05) dosages of A. membranaceus and medium (P < 0.01) and high dosages (P < 0.01) of P. notoginseng respectively as compared with control group. CONCLUSION: A. membranaceus and P. notoginseng can promote the transformation and proliferation of EPC. PMID- 16468375 TI - [Effect of different parts of Portulace oleracea on the levels of TNF-alpha and IL-6 in the supernatant of cultured adipose cell]. AB - OBJECTIVE: To explore the effects of drug-carried serum of the different parts of Portulace olerace on cytokine TNF-alpha and IL-6 secreted by adipose cell in vitro. METHOD: Models of adipose cell were established by Rodbell method. Using the method of seropharmacology, the drug-carried serum of the different parts of P. olerace were prepared. The cell viability of each group was tested by. Methy thiazolyl tetrazolium (MTT) assay. The levels of TNF-alpha and IL-6 in the supernatant of cultured adipose cell were assayed by RIA. RESULT: MTT assay results showed the cell viability of normal serum group was significantly higher than that of high lipid serum( P < 0.05). Compared with the high lipid serum group, the cell viability of the drug-carried serum groups in 40% and 20% concentration were significantly increased( P < 0.05). The high lipid serum had a better effect on increasing the levels of TNF-alpha and IL-6 than the normal serum group (P < 0.01). Expect the drug-carried serum of P. olerace low dose group in 20% concentration, each drug-carried serum group could markedly lower the levels of TNF-alpha (P < 0.05 or P < 0.01). Each drug-carried serum group in 40% concentration and the drug-carried serum P. olerace MS high dose group in 20% concentration could markedly lower the levels of IL-6 (P < 0.05 or P < 0.01). CONCLUSION: The drug-carried serum of P. olerace and its different parts act on adipose cell damaged by the high lipid serum, significantly increas the cell viability in the groups in 40% and 20% concentration, and improve the disorder of lipid in differeut degrel by lowering the levels of TNF-alpha and IL-6 that adipose cell secreted in vitro. PMID- 16468376 TI - [Protective effects of Huanglian Jiedu decoction on vascular endothelial function in type 2 diabetic rats]. AB - OBJECTIVE: To investigate the effects of Huanglian Jiedu (HLJD) decoction on vascular endothelial function in type 2 diabetic rats and explore the prophylactic and therapeutic significance and pharmacological mechanism of HLJD decoction in type 2 diabetic angiopathic complication. METHOD: The murine type 2 diabetes models were induced by the intravenous injection of a small dose of streptozotocin plus high fat and high caloric laboratory chow. Then modeled diabetic animals were divided into model group, HLJD group, and aspirin group. Normal ratsfed with routine chow were designated as normal group. The oral glucose tolerance test (OGTT) were performed in all animals, 9 weeks after treatment, the changes of murine body weights and levels of fasting blood glucose (FBG), serum total cholesterol (TC), triglycerides (TG), high density lipoprotein cholesterol(HDL-C), fasting serum insulin (FINS), serum nitric oxide, plasma endothelin, angiotensin II and von Willebrand Factor (vWF) were determined 10 weeks after treatment. RESULT: Compared with model group, the result of OGTT of HLJD group was improved. The levels of the body weights, TC, TG and ET in HLJD group weredecreased (P < 0.05). The levels of FBG,INS, AngII and vWFwere significantly decreased (P < 0.01), and the levels of HDL-C and NO were obviously increased (P < 0.05), as compared with those in model group. Furthermore. The levels of FBG was lower in HLJD group than in aspirin group (P < 0.05), and the improvement of TG, HDL-C,NO, AngII, vWF levels in HLJD group was more greatly than that in aspirin group, but there was not significant difference between two groups (P > 0.05). CONCLUSION: It is suggested by the present results that HLJD decoction could protect vascular endothelium from early damage in type 2 diabetes. The protective effects of HLJD on endothelium might be related to its ability of reducing the blood glucose, adjusting plasma lipids profiles, improving insulin resistance, antagonizing inflammatory mediators and inducing endothelium-dependent vascular relaxation. PMID- 16468377 TI - [Speciation analysis of arsenic in traditional Chinese medicine]. AB - Some of traditional Chinese medicine (TCM) contain arsenide, such as realgar. The total amount of arsenic in the TCM exceeds the limits according to related regulations. But the roles of arsenic in TCM or its side-effects depend on its species existed in those therapies, not the total amount of arsenic. Therefore, in recent years, the analysis of arsenic in TCM focuses on the species of arsenic. The present paper summarized some methods and techniques in the speciation analysis of arsenic in TCM, in order that optimal methods can be chosen and the roles of arsenic could be evaluated properly. PMID- 16468378 TI - [Research progress on oral prolonged-release preparation of traditional Chinese medicine]. AB - This paper reviews the lastest progress on oral prolonged-release preparation of traditional Chinese medicine. Four materials, include component, effective parts, single drug, and compound drugs of traditiong Chinese medicine, have been used to produce oral prolonged-release preparation. The main contents are study of preparation and evaluation of in vitro release. There are also some research works on integrative evaluation, pharmacokinetics and pharmacological activity of the prolonged-release preparation. It believes that the study on oral prolonged release preparation of traditional Chinese medicine will have good prospect. PMID- 16468379 TI - [Study progress in reversing multidrug resistance to breast cancer with Chinese herbs]. AB - Multidrug resistance of breast cancer is one of the most causes of failure in clinical chemotherapy. It is important to find out some safe and effective drugs to reserve multidrug resistance to breast cancer. The effect of some herbs had been identified in vitro. This article mainly reviewed the research progress in reversing multidrug resistance to breast cancer with Chinese herb. If the herb's effects and safety can be testified in vivo by further research, it will be effectively applied in clinic. PMID- 16468380 TI - Acetaminophen forms toxics during chlorination. PMID- 16468381 TI - Engineered wetlands may pose risks to wildlife. PMID- 16468382 TI - Lead a hazard in post-Katrina sludge. PMID- 16468383 TI - Household cleaners can remove lead dust. PMID- 16468384 TI - Acid rain in China. PMID- 16468385 TI - Including mixtures in the determination of water quality criteria for herbicides in surface water. AB - Monitoring programs throughout America and Europe have demonstrated the common occurrence of herbicides in surface water. Nevertheless, mixtures are rarely taken into account in water quality regulation. Taking mixtures into account is only feasible if the water quality criteria (WQC) of the single compounds are derived by a common and consistent methodology, which overcomes differences in data quality without settling on the lowest common denominator but making best use of all available data. In this paper, we present a method of defining a risk quotient for mixtures of herbicides with a similar mode of action (RQm). Consistent and comparable WQC are defined for single herbicides as a basis for the calculation of the RQm. Derived from the concentration addition model, the RQm can be expressed as the sum of the ratios of the measured environmental concentration and the WQC for each herbicide. The RQm should be less than one to ensure an acceptable risk to aquatic life. This approach has the advantage of being easy to calculate and communicate, and is proposed as a replacement for the current limit of 0.1 microg/L for herbicides in Switzerland. We illustrate the proposed approach on the example of five commonly applied herbicides (atrazine, simazine, terbuthylazine, isoproturon, and diuron). Their risk profile, i.e., the RQm as a function of time for one exemplary river, clearly shows that the single compounds rarely exceeded their individual WQC. However, the contribution of peaks of different seasonally applied herbicides, whose application periods partially overlap, together with the continuously emitted herbicides from nonagricultural use, results in the exceedance of the RQm threshold value of one upon several occasions. PMID- 16468386 TI - Future scenarios for a sustainable water sector: a case study from Switzerland. AB - Uncertainties about the long-term prospects of urban water management systems have increased substantially over the past decade due to an increasing variety of regulations, technologies, and demand structures. In Switzerland, this uncertainty is mirrored by growing difficulties of utility managers and (waste)water scientists to agree on shared strategies: Water professionals demand support for pressing management problems, while researchers fundamentally question the longer-term sustainability of the established water management system. To reestablish shared orientation, we conducted a foresight study for the Swiss (waste)water sector in 2004. Based on interviews with 29 experts from Swiss water management and research to collect 56 drivers of change, a team of 17 experts developed three scenarios: (A) regional mergers of water utilities leading to enhanced professionalism in the sector, (B) consequent material flows management leading to a radically restructured urban water management system, and (C) generalized financial crisis leading to a breakdown of centralized utility services. These scenarios helped identifying shared research priorities. We conclude that scenario analysis is a powerful tool for framing long-term strategies, defining priorities, and integrating different interests in the multidisciplinary contexts of sustainability science, which are marked by high uncertainties and concern a wide range of stakeholder groups. PMID- 16468387 TI - Eleven-year field study of Pu migration from Pu III, IV, and VI sources. AB - Understanding the processes controlling Pu mobility in the subsurface environment is important for estimating the amount of Pu waste that can be safely disposed in vadose zone burial sites. To study long-term Pu mobility, four 52-L lysimeters filled with sediment collected from the Savannah River Site near Aiken, South Carolina were amended with well-characterized solid Pu sources (PuIIICl3, PuIV(NO3)4, PuIV(C2O4)2, and PuVIO2(NO3)2) and left exposed to natural precipitation for 2-11 years. Pu oxidation state distribution in the Pu(III) and Pu(IV) lysimeters sediments (a red clayey sediment, pH = 6.3) were similar, consisting of 0% Pu(III), >92% Pu(IV), 1% Pu(V), 1% Pu(VI), and the remainder was a Pu polymer. These three lysimeters also had near identical sediment Pu concentration profiles, where >95% of the Pu remained within 1.25 cm of the source after 11 years; the other 5% of Pu moved at an overall rate of 0.9 cm yr( 1). As expected, Pu moved more rapidly through the Pu(VI) lysimeter, at an overall rate of 12.5 cm yr(-1). Solute transport modeling of the sediment Pu concentration profile data in the Pu(VI) lysimeter indicated that some transformation of Pu into a much less mobile form, presumably Pu(IV), had occurred during the course of the two-year study. This modeling also supported previous laboratory measurements showing that Pu(V) or Pu(VI) reduction was 5 orders of magnitude faster than corresponding Pu(III) or Pu(IV) oxidation. The slow oxidation rate (1 x 10(-8) hr(-1); t1/2 = 8000 yr) was not discernible from the Pu(VI) lysimeter data that reflected only two years of transport butwas readily discernible from the Pu(III) and Pu(IV) lysimeter data that reflected 11 years of transport. PMID- 16468388 TI - Brominated flame retardants in polar bears (Ursus maritimus) from Alaska, the Canadian Arctic, East Greenland, and Svalbard. AB - Polybrominated diphenyl ethers (PBDEs) were determined in adipose tissue of adult and subadult female polar bears sampled between 1999 and 2002 from sub populations in Arctic Canada, eastern Greenland, and Svalbard, and in males and females collected from 1994 to 2002 in northwestern Alaska. Only 4 congeners (BDE47, 99, 100, and 153) were consistently identified in all samples. BDE47 was the major PBDE congener representing from 65% to 82% of the sum (sigma) PBDEs. Age was not a significant covariate for individual PBDEs or sigmaPBDE. Higher proportions of BDE 99, 100, and 153 were generally found in samples from the Canadian Arctic than from Svalbard or the Bering-Chukchi Sea area of Alaska. Geometric mean sigmaPBDE concentrations were highest for female polar bear fat samples collected from Svalbard (50 ng/g lipid weight (lw)) and East Greenland (70 ng/g lw). Significantly lower sigmaPBDE concentrations were found in fat of bears from Canada and Alaska (means ranging from 7.6 to 22 ng/g lw). PMID- 16468389 TI - Predicting emissions of SVOCs from polymeric materials and their interaction with airborne particles. AB - A model that predicts the emission rate of volatile organic compounds (VOCs) from building materials is extended and used to predict the emission rate of semivolatile organic compounds (SVOCs) from polymeric materials. Reasonable agreement between model predictions and gas-phase di-2-ethylhexyl phthalate (DEHP) concentrations is achieved using data collected in a previous experimental study that measured emissions of DEHP from vinyl flooring in two very different chambers. While emissions of highly volatile VOCs are subject to "internal" control (the material-phase diffusion coefficient), emissions of the very low volatility SVOCs are subject to "external" control (partitioning into the gas phase, the convective mass-transfer coefficient, and adsorption onto interior surfaces). The effect of SVOCs partitioning onto airborne particles is also examined. The DEHP emission rate is increased when the gas-phase concentration is high, and especially when partitioning to the airborne particles is strong. Airborne particles may play an important role in inhalation exposure as well as in transporting SVOCs well beyond the source. Although more rigorous validation is needed, the model should help elucidate the mechanisms governing emissions of phthalate plasticizers, brominated flame retardants, biocides, and other SVOCs from a wide range of building materials and consumer products. PMID- 16468390 TI - Microscopically focused synchrotron X-ray investigation of selenium speciation in soils developing on reclaimed mine lands. AB - Chemical speciation determines Se solubility and therefore its bioavailability and potential for transport in the environment. In this study we investigated the speciation of Se in soil developed on reclaimed mine sites in the U.S. Western Phosphate Resource Area (WPRA) using micro-X-ray absorption near-edge structure (micro-XANES) spectroscopy and micro-X-ray fluorescence (micro-XRF) mapping. Selenium was nonuniformly distributed in the soils and positively correlated with Fe, Mn, Cu, Zn, and Ni. Sixteen points of interest (POI) from three soil samples were analyzed with micro-XANES spectroscopy. The XANES data indicated that Se is present in the soils in at least three oxidation states, Se(-II, 0), Se(IV), and Se(VI). Selenides or elemental Se dominated 7 of the 16 POI. Selenate was the dominant species at only one of the POI. The remaining eight POI were composed of both Se(IV) and Se(VI), with minor Se(-II, 0) contributions. The results of this research suggest that the reduced Se species in the soil parent material are oxidizing to Se(VI), one of the more mobile species of Se in the environment. This information can be used to better predict and manage Se availability in soils. PMID- 16468391 TI - Assessment of pathogens and toxicants in New Orleans, LA following Hurricane Katrina. AB - Storm surge associated with Hurricane Katrina and the breach of levees protecting New Orleans, Louisiana allowed floodwaters from Lake Pontchartrain to inundate 80% of the city. Environmental samples were collected during September 16-18, 2005 to determine immediate human and wildlife health hazards from pathogens and toxicants in the floodwaters. Baseline information on potential long-term environmental damage resulting from contaminants in water and sediments pumped into Lake Pontchartrain was also collected. Concentrations of aldrin, arsenic, lead, and seven semivolatile organic compounds in sediments/soils exceeded one or more United States Environmental Protection Agency (USEPA) thresholds for human health soil screening levels and high priority bright line screening levels. High numbers of Aeromonas spp., pathogenic Vibrio spp., and other coliform bacteria were found in floodwater samples. Alligator and snake tissues did not contain excessive toxicant concentrations. Initial findings suggest numerous environmental contaminants are present in New Orleans and support the need for further evaluation of the extent of those threats. PMID- 16468392 TI - Chemical loading into surface water along a hydrological, biogeochemical, and land use gradient: a holistic watershed approach. AB - Identifying the sources and impacts of organic and inorganic contaminants at the watershed scale is a complex challenge because of the multitude of processes occurring in time and space. Investigation of geochemical transformations requires a systematic evaluation of hydrologic, landscape, and anthropogenic factors. The 1160 km2 Boulder Creek Watershed in the Colorado Front Range encompasses a gradient of geology, ecotypes, climate, and urbanization. Streamflow originates primarily as snowmelt and shows substantial annual variation. Water samples were collected along a 70-km transect during spring runoff and base-flow conditions, and analyzed for major elements, trace elements, bulk organics, organic wastewater contaminants (OWCs), and pesticides. Major element and trace-element concentrations were low in the headwaters, increased through the urban corridor, and had a step increase downstream from the first major wastewater treatment plant (WWTP). Boron, gadolinium, and lithium were useful inorganic tracers of anthropogenic inputs. Effluent from the WWTP accounted for as much as 75% of the flow in Boulder Creek and was the largest chemical input. Under both hydrological conditions, OWCs and pesticides were detected in Boulder Creek downstream from the WWTP outfall as well as in the headwater region, and loads of anthropogenic-derived contaminants increased as basin population density increased. This report documents a suite of potential endocrine-disrupting chemicals in a reach of stream with native fish populations showing indication of endocrine disruption. PMID- 16468393 TI - Transformation of pyrene in aqueous chlorination in the presence and absence of bromide ion: kinetics, products, and their aryl hydrocarbon receptor-mediated activities. AB - To assess the endocrine-disrupting activity stemming from the presence of pyrene in drinking water, the kinetics of chlorination of pyrene was investigated at room temperature, the products of its aqueous chlorination with and without bromide ion were identified, and their aryl hydrocarbon receptor (AhR)-mediated activities were determined. It was found that the presence of bromide ion greatly promoted the reaction rate of chlorination of pyrene accompanied with the formation of brominated products. While the main product was 1-Cl-pyrene without the addition of bromide ion, di-Br-pyrene and 1-Br-pyrene became the main products in the presence of bromide ion. GC-MS and NMR analysis identified three structures of dibromopyrene in chlorination with the addition of bromide ion as 1,3-di-Br-pyrene, 1,6-di-Br-pyrene, and 1,8-di-Br-pyrene, and their molar ratio was determined to be approximately 0.3:1:1. Finally, 1-Br-pyrene, 1,3-di-Br pyrene, a mixture of 1,6-di-Br-pyrene and 1,8-di-Br-pyrene (di-Br-pyrene), 1-Cl pyrene, and a mixture of 1,6-di-Cl-pyrene and 1,8-di-Cl-pyrene (di-Cl-pyrene) were fractionated by HPLC, and their AhR-mediated activities were assessed by a yeast assay. It was found that the effective molar concentrations (or mass concentration) showing half-maximal transcriptional response, EC50, for pyrene, 1 Br-pyrene, 1-Cl-pyrene, di-Cl-pyrene, and di-Br-pyrene were 5632 (1.14), 3089 (0.86), 1942 (0.46), 597.2 (0.21), and 147.3 (0.04) nM (mg/L), respectively. PMID- 16468394 TI - Pesticide sorption and desorption by lignin described by an intraparticle diffusion model. AB - Lignin was used as a model compound for soil organic matter to gain insight into the mechanisms that control the kinetics of pesticide sorption and desorption. Hydrolytic lignin was immobilized in a matrix of alginate gel, and sorption desorption experiments were undertaken with isoproturon. Sorption increased with time and was close to equilibrium after 14 days. Desorption was measured after sorption for different time intervals and for a number of successive desorption steps of different lengths. The results showed strong differences between the sorption and desorption isotherms. The ratio of sorbed to dissolved pesticide approached and even exceeded the equilibrium ratio, depending on the number of desorption steps and the length of each equilibration period. A numerical diffusion model was developed to describe radial diffusion into the lignin particles in combination with Freundlich sorption inside the particles. Key model parameters were adjusted to fit the sorption data, and the same parameters were then used to predict stepwise desorption. Desorption was well described by the model, which suggests that sorption and desorption were driven by the same mechanism and occurred at the same rate. The observed difference between the sorption and desorption isotherms could be fully explained by the nonattainment of equilibrium due to slow diffusion into and out of the lignin particles. PMID- 16468395 TI - Comparative evaluation of the fate of disinfection byproducts at eight aquifer storage and recovery sites. AB - Despite the growth in aquifer storage and recovery (ASR) as a technique for the provision of potable water supplies, quantitative data on the fate of disinfection byproducts that may be present in the injected water remain rare. This study evaluates the data from eight ASR sites in Australia and the United States that cover a wide range of source water compositions, hydrogeological environments, and operating conditions. Rates of attenuation and formation of trihalomethanes (THMs) in groundwater were determined using analytical techniques that took dilution effects into account. Half-lives varied by more than 2 orders of magnitude (e.g., <1 to >120 days for total THMs) and were both compound- and site- specific. Chloroform was most persistent, and more highly brominated compounds tended to be less persistent, as has generally been found. For any particular THM compound, much of the variability could be explained by contrasts in geochemical conditions within the aquifer since microbial degradation is the primary mechanism for THM attenuation. As such, bounds on the half-life were defined according to the redox state of the groundwater. In situ formation of some THMs in the aquifer after injection was directly observed at a number of sites, and was predicted to have taken place at all sites. The variance in formation estimates was large between the different methods used. Formation may be more common than previously thought because of the low frequency of groundwater sampling after injection and concomitant attenuation and mixing. PMID- 16468396 TI - Replicability of bacterial communities in denitrifying bioreactors as measured by PCR/T-RFLP analysis. AB - Bioreactors hold great promise for treating graywater in an advanced life support system for space applications. However, questions remain regarding the reproducibility and reliability of biological systems for long-term use. Although there have been numerous studies on ground-based biological systems, most studies focus on a single reactor or a simple (single carbon) waste stream. There have been very few studies on microbial communities in replicate reactors using a nonsterile, complex waste stream. In this report, we describe the characterization of five replicate denitrifying reactors receiving a complex feed, including urine and limb washes from donors at Johnson Space Center over a 100-day period. Denitrifying conditions were employed because of the ease in adding a terminal electron acceptor to the bioreactor. Bacterial populations were tracked by 16S rRNA and nosZ genes T-RFLP analysis to target the total and denitrifying microbial communities. The results demonstrated reproducible biological communities with nearly identical performance that slowly changed with time and exhibited low variability with respect to the bacterial community (T RFLP peak area) in all reactors. These results suggest that, when designed for replication, bioreactors are not stochastic systems exhibiting chaotic behavior, but are biological systems that can be highly reproducible and reliable. PMID- 16468397 TI - Transformation of acetaminophen by chlorination produces the toxicants 1,4 benzoquinone and N-acetyl-p-benzoquinone imine. AB - The reaction of the common pain reliever acetaminophen (paracetamol, 4 acetamidophenol) with hypochlorite was investigated over time under conditions that simulate wastewater disinfection. Initially, the reaction was studied in pure water at neutral pH (7.0), a range of reaction times (2-90 min), and a molar excess of hypochlorite (2-57 times) relative to the acetaminophen concentration. The reaction was monitored using reversed-phase liquid chromatography (LC) with ultraviolet absorbance, electrochemical, and mass spectrometric detection. At 1 micromol/L (150 ppb) and 10 micromol/L (1.5 ppm) levels, acetaminophen readily reacted to form at least 11 discernible products, all of which exhibited greater LC retention than the parent. Two of the products were unequivocally identified as the toxic compounds 1,4-benzoquinone and N-acetyl-p-benzoquinone imine (NAPQI), which is the toxicant associated with lethality in acetaminophen overdoses. With a hypochlorite dose of 57 micromol/L (4 ppm as Cl2), 88% of the acetaminophen (10 micromol/L initial) was transformed in 1 h. The two quinoidal oxidation products 1,4-benzoquinone and NAPQI accounted for 25% and 1.5% of the initial acetaminophen concentration, respectively, at a 1 h reaction time. Other products that were identified included two ring chlorination products, chloro-4 acetamidophenol and dichloro-4-acetamidophenol, which combined were approximately 7% of the initial acetaminophen concentration at 1 h. The reaction was also studied in wastewater, where similar reactivity was noted. These results demonstrate that acetaminophen is likely to be transformed significantly during wastewater chlorination. The reactivity of the chlorine-transformation products was also studied with sulfite to simulate dechlorination, and 1,4-benzoquinone and NAPQI were completely reduced. PMID- 16468399 TI - Activity of Desulfitobacterium sp. strain Viet1 demonstrates bioavailability of 2,4-dichlorophenol previously sequestered by the aquatic plant Lemna minor. AB - Aquatic plants take up and sequester organic contaminants such as chlorophenols through incorporation in cell wall materials and storage in vacuoles. The ultimate fate of plant-sequestered chlorophenols, however, remains unclear. This research investigated 2,4-dichlorophenol (2,4-DCP) sequestration by the aquatic plant Lemna minor and evaluated contaminant release and bioavailability after plant death and cellular disruption. 14C-labeled 2,4-DCP was used to establish that contaminant removed from the aqueous phase was retained internal to L. minor. An assay with Desulfitobacterium sp. strain Viet1 was used to assess the readily bioavailable fraction of plant-sequestered 2,4-DCP and plant metabolites of 2,4-DCP. In plant-free systems, strain Viet1 dechlorinated 2,4-DCP to stoichiometric amounts of 4-chlorophenol (4-CP) as a stable and quantifiable end product. Anaerobic microcosms containing inactivated L. minor, which had accumulated 3.8 micromol of 2,4-DCP equivalents/g of plant material (fresh weight) during a preceding aerobic exposure, were inoculated with strain Viet1. After 118 d of incubation with strain Viet1, 43.5% (+/-1.4%) of the contaminant was recovered as 4-CP, indicating a large portion of plant-sequestered 2,4-DCP was bioavailable for dechlorination by strain Viet1. In contrast, 4-CP formation was not observed in autoclaved microcosms, and only 26.1% (+/-1.0%) of plant sequestered 2,4-DCP was recovered in the aqueous phase. These findings demonstrate contaminant cycling between plants and microorganisms, and emphasize that understanding the mechanisms and pathways of contaminant sequestration by plants is critical for predicting long-term contaminant fate. PMID- 16468398 TI - Oral exposure of PBDE-47 in fish: toxicokinetics and reproductive effects in Japanese Medaka (Oryzias latipes) and fathead minnows (Pimephales promelas). AB - The toxicokinetics of 2,2,4,4-tetrabromodiphenyl ether (PBDE-47) was studied in the Japanese Medaka (Oryzias latipes) after a single oral exposure followed by termination at specific time points. The effects of repeated oral exposure to PBDE-47 on reproductive performance was assessed using a pair breeding experimental design with fathead minnows (Pimephales promelas) given daily PBDE 47 exposures for 25 days, during which fecundity was measured as an indicator of reproductive performance. Medaka and fathead minnows were orally exposed to PBDE 47 by bioencapsulation in brine shrimp, Artemia sp. In the medaka studies, measurable levels of PBDE-47 were detected in the carcass within 0.25 h with peak levels occurring at 8 h. The body levels of PBDE-47 slowly declined and were still 25% of peak levels at 624 h after dosing. Assimilation of the bioencapsulated dose was at least 80% and may well approach 100%. The PBDE-47 concentration-time profile was fitted to a one-compartment clearance-volume toxicokinetic model and the model-predicted value for elimination half-life was determined to be 281 h and the first-order absorption rate constant was Ka = 0.26 hr(-1). In the fathead minnow study, egg laying in the PBDE-treated breeding pairs stopped after 10 days. The condition factor of PBDE-treated males was significantly reduced (P <0.011) compared with control males, whereas no significant difference was observed in females. Histological examination revealed a greater than 50% reduction in mature sperm in PBDE-47 exposed minnows compared to controls. Collectively, these results suggest PBDE-47 is selectively toxic to sexually mature male fathead minnows. PMID- 16468400 TI - Prediction of partitioning between complex organic mixtures and water: application of polyparameter linear free energy relationships. AB - Equilibrium partitioning between nonaqueous phase liquids (NAPLs) and water is a governing process for contaminants leaching from NAPLs. Conventional prediction methods, such as Raoult's law and single-parameter linear free energy relationship (SP-LFER), are inaccurate for compounds with polar functional groups. Therefore, this study introduces a polyparameter linear free energy relationship (PP-LFER) approach as a more general tool to predict NAPL-water partitioning coefficients. Our approach was evaluated using 441 experimental partitioning data from 30 references. Experimental fuel-water partitioning coefficients were generally well reproduced by existing PP-LFERs for pure solvents using either a volume-fraction weighted sum of partitioning coefficients K (linear model, R2 = 0.983, root-mean-squared error [rmse] = 0.23) or a volume fraction weighted sum of log K (log linear model, R2 = 0.976, rmse = 0.28). Using the linear model, estimations were, in most cases, within a factor of 2 from the experimental values, regardless of the type of compounds and the presence of a fuel additive. In contrast, the log linear model considerably underestimated partitioning coefficients in the presence of strong solute-solvent hydrogen bonding. For coal tar-water partitioning coefficients (Kcoal tar/w), new PP-LFER equations were calculated based on experimental log Kcoal tar/w values of 35 compounds. The resulting regression equation was log Kcoal tar/w = 0.40(+/-0.33) + 0.34(+/-0.32)E+ 0.61(+/-0.57)S-0.55-(+/-0.61)A-5.07(+/-0.61)B + 3.22(+/-0.35)V with the rmse equal to 0.21, where E, S, A, B, and Vare Abraham's solute descriptors. Partitioning coefficients for phenol and alcohols, calculated by the above equation, were much closer to the experimental values than to those estimated by the SP-LFER approach with octanol-water partitioning coefficients. The values of the coefficients also provide insight into the properties of coal tar in terms of molecular interactions with solutes. Consequently, using the approaches presented in this study, complex organic mixture-water partitioning coefficients of a wide range of organic compounds with varying polarity can be reasonably estimated. PMID- 16468401 TI - Degradation of chlorpyrifos in aqueous chlorine solutions: pathways, kinetics, and modeling. AB - Chlorpyrifos (CP) was used as a model compound to develop experimental methods and prototype modeling tools to forecast the fate of organophosphate (OP) pesticides under drinking water treatment conditions. CP was found to rapidly oxidize to chlorpyrifos oxon (CPO) in the presence of free chlorine. The primary oxidant is hypochlorous acid (HOCl), kr = 1.72 (+/-0.68) x 10(6) M(-1)h(-1). Thus, oxidation is more rapid at lower pH (i.e., below the pKa of HOCl at 7.5). At elevated pH, both CP and CPO are susceptible to alkaline hydrolysis and degrade to 3,5,6-trichloro-2-pyridinol (TCP), a stable end product. Furthermore, hydrolysis of both CP and CPO to TCP was shown to be accelerated in the presence of free chlorine by OCl-, kOCl,CP = 990 (+/-200) M(-1)h(-1) and kOCl,CPO = 1340 (+/-110) M(-1)h(-1). These observations regarding oxidation and hydrolysis are relevant to common drinking water disinfection processes. In this work, intrinsic rate coefficients for these processes were determined, and a simple mechanistic model was developed that accurately predicts the temporal concentrations of CP, CPO, and TCP as a function of pH, chlorine dose, and CP concentration. PMID- 16468402 TI - NMR investigation of the behavior of an organothiophosphate pesticide, methyl parathion, sorbed on clays. AB - The decomposition of methyl parathion (an organothiophosphate pesticide) sorbed on partially hydrated kaolin and montmorillonite clays (in Ca2+, Cu2+, Zn2+, and Al3+ forms) at high concentration (typically 1-10 wt %) has been examined by nuclear magnetic resonance (NMR), using solid-state 31P NMR (based on magic-angle spinning and cross polarization or direct polarization) and liquid-state 31P NMR of DMSO and acetone extracts. The results indicate that methyl parathion is initially physisorbed, appearing by solid-state 31P NMR to exhibit substantial molecular-level motion. The signals due to unreacted methyl parathion diminish and are replaced by new 31P NMR peaks resulting from hydrolysis, isomerization, and oxidation reactions over periods ranging from hours to years. 31P NMR characteristics indicate that these decomposition products are much more tightly bound to the clay than is methyl parathion. Methyl parathion decomposition is most effectively catalyzed by partially hydrated Cu(II)- and Al-montmorillonites (but with different product distributions); Ca-montmorillonite and kaolin were least effective. PMID- 16468404 TI - Potential of degradable organic chemicals for absolute and relative enrichment in the Arctic. AB - Model simulations of the fate of numerous hypothetical substances in the global environment can provide considerable insight into how an organic chemical's degradability and partitioning properties influence its absolute and relative Arctic enrichment behavior, as quantified by the Arctic Contamination Potential. For substances that degrade faster in water than in soil, but are quite persistent in the atmosphere, highest Arctic contamination is expected to occur if the substances have intermediate volatility and high hydrophobicity. Organic substances that are degradable in the atmosphere can still accumulate in the Arctic if they are soluble and highly persistent in water. These latter substances, which reach the Arctic in the ocean, also show the highest potential for relative enrichment in the Arctic, i.e., high amounts in northern high latitudes relative to the amounts in the total global environment. Beyond a threshold persistence in surface media of the order of several months to a year, chemical degradability leads to further relative enrichment. This is because only chemicals that are sufficiently long-lived get transferred to polar regions and once there can persist longer than at lower latitudes. The model simulations can inform the search for new potential Arctic contaminants, and can highlight combinations of properties which should be avoided in high production volume chemicals with the potential for environmental release. Three categories of organic substances are singled out for troublesome combinations of persistence, distribution, and potential bioaccumulation characteristics, only one of which contains "classical" Arctic POPs. Examples of potential Arctic contaminants within each of these categories are named. PMID- 16468403 TI - Analysis of nonylphenol: advances and improvements in the immunochemical determination using antibodies raised against the technical mixture and hydrophilic immunoreagents. AB - The development of an enzyme-linked immunosorbent assay (ELISA) for the detection of technical nonylphenol (NP) is reported. The preparation of specific antibodies has been addressed using an immunizing hapten with a four-carbon atom spacer arm placed at the ortho position that preserves both the hydroxyl group and the complexity of the branched nonyl chain mixture of the technical NP. The synthesis of the immunizing hapten 5-(2-hydroxy-5-nonylphenyl)-pentanoic acid has been accomplished through a four-step synthetic pathway using the NP commercial technical mixture as the starting material. Three types of competitor haptens have also been prepared depending on the location of the spacer arm: in ortho position to the phenol group (type A), attached to the oxygen atom (type B), and in para position, substituting the nonyl chain (type C). Drawbacks produced by the hydrophobicity of the NP or of the hapten derivatives have been circumvented by using a highly hydrophilic carrier molecule such as a high-molecular-weight aminodextran as a coating support for antigen in an indirect ELISA format. A reproducible and sensitive indirect competitive ELISA has been finally obtained, reaching a limit of detection of 2.3 +/- 0.9 microg L(-1) and an IC50 value of 29 +/- 5 microg L(-1) (both N = 16). A coefficient of variation of 11% for assays performed on different days (N = 5; IC50 = 30 +/- 3 microg L(-1)) demonstrates the assay reproducibility. The assay also recognizes the nonylphenol polyethoxylates to a different degree depending on the length of the ethoxylate chain. Recovery values in the range between 96 and 100% have been obtained using spiked blind aqueous samples although the sample preparation procedure used has been shown to have a great influence on the method accuracy. A preliminary evaluation of the analytical protocol established has been performed using real water samples. PMID- 16468405 TI - Detection of marine toxins, brevetoxin-3 and saxitoxin, in seawater using neuronal networks. AB - There is a need for assay systems that can detect known and unanticipated neurotoxins associated with harmful algal blooms. The present work describes our attempt to monitor the presence of brevetoxin-3 (PbTx-3) and saxitoxin (STX) in a seawater matrix using the neuronal network biosensor (NNB). The NNB relies on cultured mammalian neurons grown over microelectrode arrays, where the inherent bioelectrical activity of the network manifested as extracellular action potentials can be monitored noninvasively. Spinal cord neuronal networks were prepared from embryonic mice and the mean spike rate across the network was analyzed before and during exposure to the toxins. Extracellular action potentials from the network are highly sensitive not only to purified STX and PbTx-3, but also when in combination with matrixes such as natural seawater and algal growth medium. Detection limits for STX and PbTx-3, respectively, are 0.031 and 0.33 nM in recording buffer and 0.076 and 0.48 nM in the presence of 25-fold diluted seawater. Our results demonstrated that neuronal networks could be used for analysis of Alexandrium fundyense (STX-producer) and Karenia brevis (PbTx producer) algal samples lysed directly in the seawater-based growth medium and appropriately diluted with HEPES-buffered recording medium. The cultured network responded by changes in mean spike rate to the presence of STX-or PbTx-producing algae but not to the samples of two non-STX and non-PbTx isolates of the same algal genera. This work provides evidence that the NNB has the capacity to rapidly detect toxins associated with cells of toxic algal species or as dissolved forms present in seawater and hasthe potential for monitoring toxin levels during harmful algal blooms. PMID- 16468406 TI - Electrostatic precipitator performance and trace element emissions from two Kraft recovery boilers. AB - Fine particle emissions from combustion sources have gained attention recently due to their adverse effects on human health. The emission depends on the combustion process, fuel, and particulate removal technology. Particle concentrations at Kraft recovery boiler exits are very high, and the boilers are typically equipped with electrostatic precipitators (ESP). However, little data are available on the ESP performance in recovery boilers. Particle concentrations and size distributions were determined at two modern, operating recovery boilers. In addition, we determined the fractional collection efficiency of the ESPs by simultaneous measurements at the ESP inlet and outlet and the particulate emissions of trace metals. The particle mass concentration atthe ESP inlet was 11 24 g/Nm3 at the two boilers. Particle emissions were 30-40 mg/ Nm3 at boiler A and 12-15 mg/Nm3 at boiler B. The particle size distributions had a major particle mode at around 1 microm. These fume particles contained most of the particle mass. The main components in the particles were sodium and sulfate with minor amounts of chloride, potassium, and presumably some carbonate. The ESP collection efficiency was 99.6-99.8% at boiler A and 99.9% at boiler B. The particle penetration through the ESP was below 0.6% in the entire fume particle size range of 0.3-3 microm. Trace element emissions from both boilers were well below the limit values set by EU directive for waste incineration. PMID- 16468407 TI - Removal of lead contaminated dusts from hard surfaces. AB - Government guidelines have widely recommended trisodium phosphate (TSP) or "lead specific" cleaning detergents for removal of lead-contaminated dust (LCD) from hard surfaces, such as floors and window areas. The purpose of this study was to determine if low-phosphate, non-lead-specific cleaners could be used to efficiently remove LCD from 3 types of surfaces (vinyl flooring, wood, and wallpaper). Laboratory methods were developed and validated for simulating the doping, embedding, and sponge cleaning of the 3 surface types with 4 categories of cleaners: lead-specific detergents, nonionic cleaners, anionic cleaners, and trisodium phosphate (TSP). Vinyl flooring and wood were worn using artificial means. Materials were ashed, followed by ultrasound extraction, and anodic stripping voltammetry (ASV). One-way analysis of variance approach was used to evaluate the surface and detergent effects. Surface type was found to be a significant factor in removal of lead (p < 0.001). Vinyl flooring cleaned better than wallpaper by over 14% and wood cleaned better than wallpaper by 13%. There was no difference between the cleaning action of vinyl flooring and wood. No evidence was found to support the use of TSP or lead-specific detergents over all purpose cleaning detergents for removal of lead-contaminated dusts. No-phosphate, non-lead-specific detergents are effective in sponge cleaning of lead contaminated hard surfaces and childhood lead prevention programs should consider recommending all-purpose household detergents for removal of lead-contaminated dust after appropriate vacuuming. PMID- 16468408 TI - An oil-absorber-bioscrubber system to stabilize biotreatment of pollutants present in waste gas. Fluctuating loads of 1,2-dichloroethane. AB - Biotreatment technologies offer a cost-effective and efficient method for dealing with point-source releases of solvents. However, a major problem hampering these technologies is the fluctuating pollutant loads, which is especially critical for inhibitory pollutants. Provision of biotreatment systems able to cope with this problem is a significant technological and environmental challenge. This study investigates the potential for an absorber to act as buffer for shock loadings of inhibitory pollutants in waste-gas streams undergoing biological treatment. 1,2 Dichloroethane (DCE) was used as an example of a toxic and inhibitory organic pollutant. The stability of a combined oil-absorber-bioscrubber (OAB) system was compared to that of a bioscrubber only (BO) system when each was subjected to shock loads of DCE. The BO system was inoculated with Xanthobacter autotrophicus strain GJ10 and was submitted to sharp, sequential pulses in DCE inlet load, which caused system instability. Complete inhibition of the BO process occurred for a 3 h DCE pulse, leading to 9125 g of DCE m(-3)bioscrubber total organic discharged (TODDCE). Following the pulse, fluorescence in situ hybridization (FISH) showed that the active strain GJ10 was effectively washed-out. In contrast, the performance of the OAB system was stable during DCE shock loads. The carbon dioxide production remained stable, and low levels of effluent DCE and total organic carbon concentrations were found. For the 3 h pulse TODDCE was only 173 g of DCE m(-3)bioscrubber, and FISH indicated that the GJ10 strain remained active. We conclude that the OAB system offers an effective solution to the biological treatment of waste-gas containing fluctuating pollutant concentrations. PMID- 16468409 TI - Accumulation of contaminants in fish from wastewater treatment wetlands. AB - Increasing demands on water resources in arid environments make reclamation and reuse of municipal wastewater an important component of the water budget. Treatment wetlands can be an integral part of the water-reuse cycle providing both water-quality enhancement and habitat functions. When used for habitat, the bioaccumulation potential of contaminants in the wastewater is a critical consideration. Water and fish samples collected from the Tres Rios Demonstration Constructed Wetlands near Phoenix, Arizona, which uses secondary-treated wastewater to maintain an aquatic ecosystem in a desert environment, were analyzed for hydrophobic organic compounds (HOC) and trace elements. Semipermeable membrane devices (SPMD) were deployed to investigate uptake of HOC. The wetlands effectively removed HOC, and concentrations of herbicides, pesticides, and organic wastewater contaminants decreased 40-99% between inlet and outlet. Analysis of Tilapia mossambica and Gambusia affinis indicated accumulation of HOC, including p,p'-DDE and trans-nonachlor. The SPMD accumulated the HOC detected in the fish tissue as well as additional compounds. Trace element concentrations in whole-fish tissue were highly variable, but were similar between the two species. Concentrations of HOC and trace elements varied in different fish tissue compartments, and concentrations in Tilapia liver tissue were greater than those in the whole organism or filet tissue. Bioconcentration factors for the trace elements ranged from 5 to 58,000 and for the HOC ranged from 530 to 150,000. PMID- 16468410 TI - Destruction of gas-phase trichloroethylene in a modified fuel cell. AB - A conventional fuel cell was used as a catalytic reactor to treat soil vapor extraction (SVE) gases contaminated with trichloroethylene (TCE). The SVE gases are fed to the cathode side of the fuel cell, where TCE is reduced to ethane and hydrochloric acid. The results obtained suggest that TCE reduction occurs by a catalytic reaction with hydrogen that is re-formed on the cathode's surface beyond a certain applied cell potential. Substantial conversion of TCE is obtained, even when competing oxygen reduction occurs in the cathode. The process has been modeled successfully by conceptualizing the flow passage in the fuel cell as a plug flow reactor. PMID- 16468411 TI - Fenton degradation of organic pollutants in the presence of low-molecular-weight organic acids: cooperative effect of quinone and visible light. AB - The influence of low-molecular-weight organic acids (LMWOAs), such as malonic acid, ethylenediaminetetraacetic acid, and oxalic acid, on the Fenton degradation of organic pollutants was examined under visible irradiation (lambda > 450 nm). The Fenton degradation of malachite green in the dark was completely blocked in the presence of LMWOAs. It was found that either visible light irradiation or the addition of hydroquinone could initiate the dye degradation, but the mineralization yield was almost zero. An important result was that the dye mineralization in the presence of LMWOAs could be achieved when both visible irradiation and hydroquinone were introduced. Similar results were obtained with colorless pollutants, such as benzyltrimethylammonium chloride and 2,4,5 trichlorophenol. We concluded that coupling visible irradiation and hydroquinone could be a strong and universal driving force in the Fenton reaction for the complete degradation and mineralization of organic pollutants, even in the presence of LMWOAs. PMID- 16468412 TI - Dioxiranes generated in situ from pyruvates and oxone as environmentally friendly oxidizing agents for disinfection. AB - Dioxiranes generated in situ from pyruvates (alpha-keto esters) and Oxone have been found to be environmentally friendly oxidizing agents for disinfection. These oxidizing agents were highly effective for destruction of various strains of bacteria, fungi, and bacterial endospores in a wide temperature range with exceptional stability. Notably, by using an aqueous solution of methyl pyruvate (1a) and Oxone/NaHCO3, complete destruction of bacteria such as Staphylococcus aureus and fungus Penicillium corylophilum was achieved within 5 min at 20 degrees C at neutral pH. Highly chemical-resistant bacterial endospores of Bacillus cereus could also be destroyed. The high antibacterial activity of 1a could be attributed to its strong electron-withdrawing alpha-ester group. PMID- 16468413 TI - Maintenance of certification for pathologists: what we know so far. PMID- 16468414 TI - Establishing a compensation model in an academic pathology department. AB - Formal compensation models can be a valuable tool in building a strong academic pathology department. A good compensation model can increase staff satisfaction and academic productivity. A critical element to developing a well-accepted model is that the model emphasizes the core values and priorities of the department. This article highlights how the Pathology Service at Massachusetts General Hospital (MGH) created a compensation model for its Anatomic Pathology and Laboratory Medicine division, from early discussions through rollout. It also examines the impact that the salary model has had at MGH in the 4 years since it was rolled out. PMID- 16468416 TI - Pharmacogenomics and clinical biomarkers in drug discovery and development. AB - The fields of pathology and clinical pharmacology are in an era of rapid evolution, reflecting continuous technological advancements in molecular diagnostics focused on improving drug efficacy and reducing toxicity. The discovery and refinement of the human genome sequence, further understanding of epigenetic events, and the expansion of proteomics research combined with emerging technologies such as functional imaging, biosensors, and sophisticated computational biology are having an unprecedented impact on the pharmaceutical industry. This review focuses on the discovery and development of DNA-based gene sequence tests and RNA-based gene expression profiles as applied to the prediction of response, resistance, and toxicity of both new and existing anticancer, cardiovascular, and anti-inflammatory drugs. This review will also consider the potential of emerging biomarkers designed to assist in the clinical development of these agents. PMID- 16468415 TI - Tumor suppressor genes in breast cancer: the gatekeepers and the caretakers. AB - Tumor suppressor genes encode for proteins whose normal function is to inhibit cell transformation and whose inactivation is advantageous for tumor cell growth and survival. A variety of mechanisms result in the inactivation of tumor suppressor genes, including intragenic mutations, chromosomal deletions, and loss of expression by methylation-mediated transcriptional silencing or increased proteolysis. Tumor suppressor genes participate in a variety of critical and highly conserved cell functions, including regulation of the cell cycle and apoptosis, differentiation, surveillance of genomic integrity and repair of DNA errors, signal transduction, and cell adhesion. Tumor suppressor functions can be separated into 2 major categories: gatekeepers and caretakers. Gatekeepers directly inhibit tumor growth or promote tumor death. Inactivation of these genes contributes directly to cancer formation and progression. Among them, the p53 gene is the most well known. Located on chromosome band 17p13, p53 encodes a 53 kd multifunctional transcription factor that regulates the expression of genes involved in cell cycle control, apoptosis, DNA repair, and angiogenesis. In breast cancer, most studies have shown that p53 mutation or down-regulation is associated with adverse prognosis. Other tumor suppressor genes of interest in breast cancer include the retinoblastoma gene (pRb), PTEN, p16, nm23, and maspin. PMID- 16468417 TI - Recent developments and selected diagnostic problems in carcinomas of the endometrium. AB - Two major types of endometrial carcinoma have been well described as differing not only in their pathogenesis but also in their behavior, therefore requiring different management. These implications make the differential diagnosis between the 2 pathogenetic types, specifically their prototypes, endometrioid and serous endometrial carcinomas, paramount. However, this differential diagnosis may be difficult owing to overlapping histologic features such as papillary architecture. To that end, the differential diagnosis of endometrial tumors with papillary morphologic features is discussed in detail. Preinvasive and early invasive serous carcinoma (endometrial intraepithelial carcinoma and minimal serous carcinoma) have been claimed to have a good prognosis if accurate staging confirmed lack of extrauterine spread. Definitions and diagnostic features of these small serous lesions are described. In addition, several recently described variants and histologic patterns of endometrial carcinoma that might present diagnostic problems, such as microglandular mucinous carcinoma, sertoliform carcinoma, carcinoma arising in an atypical polypoid adenomyoma, and endometrial carcinoma with a minimal deviation adenocarcinoma pattern of myoinvasion, are described. PMID- 16468418 TI - Cutaneous melanocytic lesions: selected problem areas. AB - Cutaneous melanocytic proliferations are diverse morphologically, and their behavioral attributes may be difficult to discern with certainty. As a consequence, diagnostic anatomic pathologists regularly encounter problems in the interpretation of such lesions. This review considers several selected issues in that subject area, including proliferative congenital nevi, architecturally disordered (dysplastic) nevi, morphologic variants of Spitz nevus, atypical lentiginous melanocytic proliferations, nevoid melanoma, diagnostically deceptive histologic variants of melanoma, "epidermotropic" metastases of melanoma, and the relationship of melanoma microstages to tumor growth phases. PMID- 16468419 TI - The primary vasculitides: a clinicopathologic correlation. AB - Primary vasculitis is the inflammation and necrosis of vessel walls not associated with infections, drugs, and autoimmune and lymphoproliferative disorders. It is important to make the correct diagnosis of different types of vasculitis, as their prognosis may be significantly different. Classification of vasculitis based on the size of the vessel is helpful, but there is often an overlap. Whereas the criteria proposed by the American College of Rheumatology are primarily clinical, the definitions set forth by the Chapel Hill Consensus Conference are based only on histologic observations. Correct diagnosis requires appropriate incorporation of the clinical history, laboratory parameters, and the histologic data. Incorporation of antineutrophil cytoplasmic antibodies in defining the pathogenesis of vasculitis has been particularly useful in diagnosing those small vessel vasculitides that are life threatening and need immediate intervention. PMID- 16468420 TI - Squamous cell carcinoma variants of the upper aerodigestive tract. AB - Squamous cell carcinoma (SCC) is by far the most common malignancy of the upper aerodigestive tract. Most conventional-type SCCs do not present the surgical pathologist with diagnostic difficulty. Certain variants, however, can histologically mimic benign and other malignant diseases. In such cases, correct diagnosis is important for prognostic and therapeutic reasons. This article discusses the clinicopathologic features of the upper aerodigestive tract variants of SCC, including verrucous carcinoma, papillary squamous cell carcinoma, basaloid squamous cell carcinoma, spindle cell carcinoma, adenosquamous carcinoma, adenoid squamous cell carcinoma, and undifferentiated carcinoma. It also discusses the differential diagnoses that must be considered and the use of ancillary testing, especially immunohistochemical analysis, for determining the correct diagnosis. PMID- 16468421 TI - "Undifferentiated" small round cell tumors of the sinonasal tract: differential diagnosis update. AB - Sinonasal tract neoplasms composed of light microscopically seemingly "undifferentiated" small round cells often generate considerable diagnostic difficulty. Although the careful review of H&E-stained sections remains of critical and central importance in this evaluation, the recent improvements in the immunohistochemical diagnostic armamentarium and molecular diagnostic techniques applicable to paraffin-embedded tissue samples may add diagnostically valuable information. Accordingly, this review will discuss the differential diagnosis of undifferentiated small blue cell tumors of the sinonasal tract based on the light microscopic and clinical features and, as needed, the results of these ancillary studies. Tumors discussed include olfactory neuroblastoma, sinonasal undifferentiated carcinoma, small cell undifferentiated (neuroendocrine) carcinoma, undifferentiated (lymphoepithelioma-like) carcinoma, malignant melanoma, pituitary adenoma, Ewing sarcoma/peripheral neuroectodermal tumor, rhabdomyosarcoma, mesenchymal chondrosarcoma, small cell osteosarcoma, synovial sarcoma, extranodal natural killer/T-cell lymphoma, nasal type, and extramedullary plasmacytoma. PMID- 16468423 TI - Virtual Jihad. PMID- 16468422 TI - Laboratory testing for B-type natriuretic peptides (BNP and NT-proBNP): clinical usefulness, utilization, and impact on hospital operations. AB - B-type natriuretic peptides (BNP, NT-proBNP) are involved in cardiovascular remodeling, fluid and electrolyte balance, and myocardial response to coronary ischemia. Recent literature has demonstrated the usefulness of natriuretic peptides in the diagnosis of congestive heart failure (CHF). Further studies also have shown that B-type natriuretic peptides (BNPs) are useful to guide therapy in patients with established CHF and might be useful to estimate prognosis in patients with CHF and in those with left ventricular dysfunction due to acute coronary syndromes. Consequently, the majority of laboratories in the United States perform testing for BNP or its inactive aminoterminal fragment, NT-proBNP Most experts agree that blood levels of BNP and NT-proBNP can be used interchangeably, although important differences have been identified. Although the clinical usefulness of BNPs has been well described, few studies have reported data on the utilization of these markers or their impact on patient outcomes and hospital operations. This review describes the biochemical characteristics, clinical usefulness, patterns of utilization, and economic impact of laboratory testing for the BNPs. PMID- 16468424 TI - T cells for brain cells. PMID- 16468425 TI - Un-killing the messenger. PMID- 16468426 TI - Half-brained schemes. PMID- 16468427 TI - Clean and green. PMID- 16468428 TI - Pumped-up performance. PMID- 16468429 TI - Modern slavery. PMID- 16468430 TI - Murdercide. PMID- 16468431 TI - Easing jitters when buildings rumble. PMID- 16468432 TI - The mystery of brown dwarf origins. PMID- 16468433 TI - Genomes for all. PMID- 16468435 TI - Innovations from a robot rally. PMID- 16468434 TI - Tsunami: wave of change. PMID- 16468437 TI - Recognition engines. PMID- 16468436 TI - The maternal brain. PMID- 16468439 TI - No more gears. PMID- 16468440 TI - Why does skin wrinkle with age? How can you slow or prevent this process? PMID- 16468438 TI - Protecting more than animals. PMID- 16468441 TI - "Ordinary people" and "death work": Palestinian suicide bombers as victimizers and victims. AB - Applying criminological/victimological concepts and theories, the study addresses the social processes involved in Palestinians' suicide terrorism and describes Palestinians' pathways to suicide bombing. The data are derived from in-depth interviews of 7 male and female Palestinians serving prison sentences in Israel for attempted suicide bombing. The social background, context, and experiences of the interviewees, including their recruitment, interactions with the organizations that produce suicide bombing, the tangible and intangible incentives and rewards that motivated them to become suicide bombers, their preparation for the mission, and the strategies employed by the organizations to sustain recruits' resolve to conform to the plan are described and analyzed. The implications of the findings for theory and public policy are drawn and discussed. PMID- 16468442 TI - PTSD symptoms among men and women survivors of intimate partner violence: the role of risk and protective factors. AB - The purpose of this cross-sectional analysis of the National Violence Against Women Survey was to characterize current symptoms of posttraumatic stress disorder (PTSD) among 185 men and 369 women survivors of intimate partner violence (IPV). In this subsample, 24% of women and 20% of men had current moderate-to-severe PTSD symptoms. PTSD scores were higher for women than men. Protective factors that appear to increase resiliency of survivors were higher education and income, being currently married, and reporting that IPV had stopped. Higher physical or psychological IPV scores, current depressive symptoms, and the survivor having left the relationship at least once were associated with risk of moderate-to-severe symptoms of PTSD. Protective factors may be used to boost resiliency of IPV survivors and reduce PTSD symptoms. PMID- 16468443 TI - Posttraumatic stress disorder symptomatology and comorbid depressive symptoms among abused women referred from emergency department care. AB - Posttraumatic stress disorder (PTSD) is common among victims of intimate partner violence (IPV) as is comorbid depression. Comorbid depression may exacerbate PTSD severity and chronicity. This study sampled female IPV victims from an urban emergency department to assess the relationship between PTSD symptomatology in the previous 12 months and current depressive symptomatology and to evaluate independent predictors of PTSD symptomatology. Half of respondents had symptoms consistent with PTSD. Those with PTSD symptomatology had significantly higher mean total depression scores and mean scores on 3 of 4 depression subscales than those without PTSD. Depressive symptomatology, being married, sexual IPV, severity of physical IPV, and partner's consumption of 5 or more alcoholic drinks per occasion at least once a month independently predicted PTSD symptomatology. Our findings underscore the important roles these factors play in IPV-related PTSD and the need for prompt identification and intervention of those at risk. PMID- 16468444 TI - Conducting research on violence in Asian American communities: methodological issues. AB - Conducting culturally competent research is a challenge as the United States becomes increasingly multicultural. When conducting research on violence in Asian American communities, researchers need to consider how culture, race, and ethnicity influence definitions of concepts, and methodological issues such as research designs, sampling, developing and translating instruments, ethical issues, recruiting research participants, supervising and training interviewers, and disseminating findings. Examples from the authors' research studies on community violence in the Khmer community, domestic violence in the Chinese American community, and dating violence in Asian American groups are extrapolated to highlight various themes. A commitment to a research program that collaborates with the community under study and cultural experts is vital at every stage of the research process. PMID- 16468445 TI - Prevalence and correlates of interpersonal violence victimization in a junior enlisted Navy cohort. AB - This study assesses the prevalence and correlates of interpersonal violence victimization in a cohort of 493 male and 220 female junior Navy personnel who participated in a cross-sectional follow-up study on health-related behaviors. Survey data were obtained during 2000 about past-year prevalence of physical violence and sexual victimization. Chi-square tests of independence and multivariable logistic regression analyses were employed to estimate prevalence rates and to assess correlates of interpersonal violence. No gender differences were found for rates of violence victimization (16.4%) or sexual victimization (4.5%). Significant correlates of interpersonal violence victimization among males were depression and tobacco use; among females, significant correlates were depression and frequent heavy drinking. Findings suggest that large numbers of young enlisted adults serving in the military may be victims of interpersonal violence. PMID- 16468446 TI - Intimate sexual victimization among women with protective orders: types and associations of physical and mental health problems. AB - Intimate sexual violence was examined among a sample of women who had recently obtained protective orders against male partners using three groups: no sexual victimization (n = 368), sexual insistence (n = 114), and threatened and/or forced sex (n = 117). Differences in childhood sexual abuse as well as types of partner psychological abuse, stalking, and severe physical violence experiences were found across the groups. Multivariate analysis showed that women with no sexual victimization had significantly fewer mental health problems than women who had experienced sexual insistence and women who had been threatened or forced to have sex. Findings from this study underscore the importance of health, mental health, and criminal justice professionals assessing for a range of sexually abusive acts when working with victims of partner violence. PMID- 16468447 TI - Psychological impact of writing about abuse or positive experiences. AB - Writing often helps people deal with trauma. To see if writing about childhood physical or sexual abuse, or positive experiences, helps, psychology undergraduates wrote for 20 minutes on 4 days about their abuse, a positive experience, or a trivial topic. Among 102 who began and 85 who completed pre-, post-, and 4-week follow-up measures of depression, anxiety, and suicidal ideas, abuse writers were more likely to discontinue, and positive writers were more depressed and anxious. Compared to pretest, all completers were less depressed, anxious, and suicidal at follow-up, but nonsignificantly different in health visits. Completers who wrote about abuse rated the study as more valuable than did those who wrote about positive experiences. College students who wrote about childhood physical or sexual abuse benefited from any type of structured writing assignment (where they interacted with a researcher and got extra credit) in terms of reduced anxiety, depression, and suicidal ideas, but they found value in writing about their trauma more than writing about innocuous topics. PMID- 16468448 TI - Super-resolution in PET imaging. AB - This paper demonstrates a super-resolution method for improving the resolution in clinical positron emission tomography (PET) scanners. Super-resolution images were obtained by combining four data sets with spatial shifts between consecutive acquisitions and applying an iterative algorithm. Super-resolution attenuation corrected PET scans of a phantom were obtained using the two-dimensional and three-dimensional (3-D) acquisition modes of a clinical PET/computed tomography (CT) scanner (Discovery LS, GEMS). In a patient study, following a standard 18F FDG PET/CT scan, a super-resolution scan around one small lesion was performed using axial shifts without increasing the patient radiation exposure. In the phantom study, smaller features (3 mm) could be resolved axially with the super resolution method than without (6 mm). The super-resolution images had better resolution than the original images and provided higher contrast ratios in coronal images and in 3-D acquisition transaxial images. The coronal super resolution images had superior resolution and contrast ratios compared to images reconstructed by merely interleaving the data to the proper axial location. In the patient study, super-resolution reconstructions displayed a more localized 18F-FDG uptake. A new approach for improving the resolution of PET images using a super-resolution method has been developed and experimentally confirmed, employing a clinical scanner. The improvement in axial resolution requires no changes in hardware. PMID- 16468449 TI - Generation and visualization of four-dimensional MR angiography data using an undersampled 3-D projection trajectory. AB - Time-resolved contrast-enhanced magnetic resonance (MR) angiography (CE-MRA) has gained in popularity relative to X-ray Digital Subtraction Angiography because it provides three-dimensional (3-D) spatial resolution and it is less invasive. We have previously presented methods that improve temporal resolution in CE-MRA while providing high spatial resolution by employing an undersampled 3-D projection (3D PR) trajectory. The increased coverage and isotropic resolution of the 3D PR acquisition simplify visualization of the vasculature from any perspective. We present a new algorithm to develop a set of time-resolved 3-D image volumes by preferentially weighting the 3D PR data according to its acquisition time. An iterative algorithm computes a series of density compensation functions for a regridding reconstruction, one for each time frame, that exploit the variable sampling density in 3D PR. The iterative weighting procedure simplifies the calculation of appropriate density compensation for arbitrary sampling patterns, which improve sampling efficiency and, thus, signal to-noise ratio and contrast-to-noise ratio, since it is does not require a closed form calculation based on geometry. Current medical workstations can display these large four-dimensional studies, however, interactive cine animation of the data is only possible at significantly degraded resolution. Therefore, we also present a method for interactive visualization using powerful graphics cards and distributed processing. Results from volunteer and patient studies demonstrate the advantages of dynamic imaging with high spatial resolution. PMID- 16468450 TI - Pinhole SPECT imaging: compact projection/backprojection operator for efficient algebraic reconstruction. AB - We describe the efficient algebraic reconstruction (EAR) method, which applies to cone-beam tomographic reconstruction problems with a circular symmetry. Three independant steps/stages are presented, which use two symmetries and a factorization of the point spread functions (PSFs), each reducing computing times and eventually storage in memory or hard drive. In the case of pinhole single photon emission computed tomography (SPECT), we show how the EAR method can incorporate most of the physical and geometrical effects which change the PSF compared to the Dirac function assumed in analytical methods, thus showing improvements on reconstructed images. We also compare results obtained by the EAR method with a cubic grid implementation of an algebraic method and modeling of the PSF and we show that there is no significant loss of quality, despite the use of a noncubic grid for voxels in the EAR method. Data from a phantom, reconstructed with the EAR method, demonstrate 1.08-mm spatial tomographic resolution despite the use of a 1.5-mm pinhole SPECT device and several applications in rat and mouse imaging are shown. Finally, we discuss the conditions of application of the method when symmetries are broken, by considering the different parameters of the calibration and nonsymmetric physical effects such as attenuation. PMID- 16468451 TI - Conductivity image reconstruction from defective data in MREIT: numerical simulation and animal experiment. AB - Magnetic resonance electrical impedance tomography (MREIT) is designed to produce high resolution conductivity images of an electrically conducting subject by injecting current and measuring the longitudinal component, Bz, of the induced magnetic flux density B = (Bx, By, Bz). In MREIT, accurate measurements of Bz are essential in producing correct conductivity images. However, the measured Bz data may contain fundamental defects in local regions where MR magnitude image data are small. These defective Bz data result in completely wrong conductivity values there and also affect the overall accuracy of reconstructed conductivity images. Hence, these defects should be appropriately recovered in order to carry out any MREIT image reconstruction algorithm. This paper proposes a new method of recovering Bz data in defective regions based on its physical properties and neighboring information of Bz. The technique will be indispensable for conductivity imaging in MREIT from animal or human subjects including defective regions such as lungs, bones, and any gas-filled internal organs. PMID- 16468452 TI - Super-resolution registration using tissue-classified distance fields. AB - We present a method for registering the position and orientation of bones across multiple computed-tomography (CT) volumes of the same subject. The method is subvoxel accurate, can operate on multiple bones within a set of volumes, and registers bones that have features commensurate in size to the voxel dimension. First, a geometric object model is extracted from a reference volume image. We use then unsupervised tissue classification to generate from each volume to be registered a super-resolution distance field--a scalar field that specifies, at each point, the signed distance from the point to a material boundary. The distance fields and the geometric bone model are finally used to register an object through the sequence of CT images. In the case of multiobject structures, we infer a motion-directed hierarchy from the distance-field information that allows us to register objects that are not within each other's capture region. We describe a validation framework and evaluate the new technique in contrast with grey-value registration. Results on human wrist data show average accuracy improvements of 74% over grey-value registration. The method is of interest to any intrasubject, same-modality registration applications where subvoxel accuracy is desired. PMID- 16468453 TI - Mammogram registration: a phantom-based evaluation of compressed breast thickness variation effects. AB - The temporal comparison of mammograms is complex; a wide variety of factors can cause changes in image appearance. Mammogram registration is proposed as a method to reduce the effects of these changes and potentially to emphasize genuine alterations in breast tissue. Evaluation of such registration techniques is difficult since ground truth regarding breast deformations is not available in clinical mammograms. In this paper, we propose a systematic approach to evaluate sensitivity of registration methods to various types of changes in mammograms using synthetic breast images with known deformations. As a first step, images of the same simulated breasts with various amounts of simulated physical compression have been used to evaluate a previously described nonrigid mammogram registration technique. Registration performance is measured by calculating the average displacement error over a set of evaluation points identified in mammogram pairs. Applying appropriate thickness compensation and using a preferred order of the registered images, we obtained an average displacement error of 1.6 mm for mammograms with compression differences of 1-3 cm. The proposed methodology is applicable to analysis of other sources of mammogram differences and can be extended to the registration of multimodality breast data. PMID- 16468454 TI - Observer efficiency in discrimination tasks simulating malignant and benign breast lesions imaged with ultrasound. AB - We investigate and extend the ideal observer methodology developed by Smith and Wagner to detection and discrimination tasks related to breast sonography. We provide a numerical approach for evaluating the ideal observer acting on radio frequency (RF) frame data, which involves inversion of large nonstationary covariance matrices, and we describe a power-series approach to computing this inverse. Considering a truncated power series suggests that the RF data be Wiener filtered before forming the final envelope image. We have compared human performance for Wiener-filtered and conventional B-mode envelope images using psychophysical studies for 5 tasks related to breast cancer classification. We find significant improvements in visual detection and discrimination efficiency in four of these five tasks. We also use the Smith-Wagner approach to distinguish between human and processing inefficiencies, and find that generally the principle limitation comes from the information lost in computing the final envelope image. PMID- 16468455 TI - Wavelet-based reconstruction for limited-angle X-ray tomography. AB - The aim of X-ray tomography is to reconstruct an unknown physical body from a collection of projection images. When the projection images are only available from a limited angle of view, the reconstruction problem is a severely ill-posed inverse problem. Statistical inversion allows stable solution of the limited angle tomography problem by complementing the measurement data by a priori information. In this work, the unknown attenuation distribution inside the body is represented as a wavelet expansion, and a Besov space prior distribution together with positivity constraint is used. The wavelet expansion is thresholded before reconstruction to reduce the dimension of the computational problem. Feasibility of the method is demonstrated by numerical examples using in vitro data from mammography and dental radiology. PMID- 16468456 TI - Parallelized Bayesian inversion for three-dimensional dental X-ray imaging. AB - Diagnostic and operational tasks based on dental radiology often require three dimensional (3-D) information that is not available in a single X-ray projection image. Comprehensive 3-D information about tissues can be obtained by computerized tomography (CT) imaging. However, in dental imaging a conventional CT scan may not be available or practical because of high radiation dose, low resolution or the cost of the CT scanner equipment. In this paper, we consider a novel type of 3-D imaging modality for dental radiology. We consider situations in which projection images of the teeth are taken from a few sparsely distributed projection directions using the dentist's regular (digital) X-ray equipment and the 3-D X-ray attenuation function is reconstructed. A complication in these experiments is that the reconstruction of the 3-D structure based on a few projection images becomes an ill-posed inverse problem. Bayesian inversion is a well suited framework for reconstruction from such incomplete data. In Bayesian inversion, the ill-posed reconstruction problem is formulated in a well-posed probabilistic form in which a priori information is used to compensate for the incomplete information of the projection data. In this paper we propose a Bayesian method for 3-D reconstruction in dental radiology. The method is partially based on Kolehmainen et al. 2003. The prior model for dental structures consist of a weighted l1 and total variation (TV)-prior together with the positivity prior. The inverse problem is stated as finding the maximum a posteriori (MAP) estimate. To make the 3-D reconstruction computationally feasible, a parallelized version of an optimization algorithm is implemented for a Beowulf cluster computer. The method is tested with projection data from dental specimens and patient data. Tomosynthetic reconstructions are given as reference for the proposed method. PMID- 16468457 TI - Modeling the amplitude statistics of ultrasonic images. AB - In this paper, a new statistical model for representing the amplitude statistics of ultrasonic images is presented. The model is called the Rician inverse Gaussian (RiIG) distribution, due to the fact that it is constructed as a mixture of the Rice distribution and the Inverse Gaussian distribution. The probability density function (pdf) of the RiIG model is given in closed form as a function of three parameters. Some theoretical background on this new model is discussed, and an iterative algorithm for estimating its parameters from data is given. Then, the appropriateness of the RiIG distribution as a model for the amplitude statistics of medical ultrasound images is experimentally studied. It is shown that the new distribution can fit to the various shapes of local histograms of linearly scaled ultrasound data better than existing models. A log-likelihood cross-validation comparison of the predictive performance of the RiIG, the K, and the generalized Nakagami models turns out in favor of the new model. Furthermore, a maximum a posteriori (MAP) filter is developed based on the RiIG distribution. Experimental studies show that the RiIG MAP filter has excellent filtering performance in the sense that it smooths homogeneous regions, and at the same time preserves details. PMID- 16468458 TI - Is our nation fully prepared to provide disaster relief to victims needing dental care? PMID- 16468459 TI - Dental insurance and clinical dental outcomes in NHANES III. AB - OBJECTIVES: The National Health and Nutrition Examination Survey (NHANES III) 1988-1994 is one of the few nationally representative data sets with information on both private dental insurance and a clinical dental exam. The objective of this analysis was to examine the possible associations between private dental insurance and clinical exam outcomes, demographic variables, and dental visits. METHODS: Using NHANES III data, analysis was limited to persons aged 20 years or older who had a dental exam and reported on their private dental insurance status. Initial analyses were based on comparisons between those with and without private dental insurance. Propensity scoring method was used to examine the effects of dental insurance on clinical exam variables. RESULTS: The percentage of individuals with private dental insurance was significantly greater among non Hispanic blacks, those with higher educational attainment, those living at/above the federal poverty level, and those with a dental visit in the past year compared to their respective counterparts. Those with untreated caries, those with a loss of attachment of greater than 4 mm, and those with 12-27 missing teeth were significantly less likely to have dental insurance (p < 0.05) than their respective counterparts. CONCLUSIONS: These results suggest that having private dental insurance is associated with better clinical oral health status. PMID- 16468460 TI - Determinants of dental referral practices among WIC nutritionists in North Carolina. AB - OBJECTIVES: To examine the effects of knowledge and confidence on dental referral practices among WIC nutritionists in North Carolina. METHODS: A questionnaire consisting of 118 Likert scale-type questions was administered to 92% of all WIC nutritionists in North Carolina (n = 324). The relationship of knowledge and confidence with frequent dental referrals was tested using logistic regression. RESULTS: Regression results found that confidence in performing oral health risk assessments (OR = 2.12; 95% CI = 1.13, 3.96), confidence in making dental referrals (OR = 3.02; 95% CI = 1.45-6.29), and confidence in expected outcomes that parents would seek dental care when advised to do so (OR = 3.11; 95% CI = 1.62, 5.97) were associated with more frequent dental referrals. CONCLUSIONS: The more confident WIC nutritionists feel about oral health, the more likely they are to make dental referrals. Screening and referral by WIC workers may benefit children by improving access to dental care, as the WIC clinic is frequently the first point of contact with a health professional. PMID- 16468461 TI - Risk assessment criteria applied to a screening exam: implications for improving the efficiency of a sealant program. AB - OBJECTIVES: The present study aimed to determine the ability of first-grade screening exam findings to predict carious lesions in permanent first molars by fourth grade. The ability to identify high-risk children would be useful in the rational allocation of limited public health resources, such as the application of sealants. METHODS: Screening exams were performed on 204 children in 1st grade and 4th grade. Analyses were conducted at both the child- and molar- levels. Cross-tabulations were used to identify the threshold dmfs + DMFS in first grade that had the highest sensitivity and highest negative predictive value for discriminating between children with and without permanent caries (1) in the entire population and (2) in subpopulations defined by race/ethnicity, gender, and language spoken at home. This threshold then was entered into logistic regression models. RESULTS: On the child and molar level, the study determined that dmfs + DMFS > 0 had the highest sensitivity (child: 69.4%, molar: 74.7%) and negative predictive value (child: 87.8%, molar: 94.4%). Using this test criterion, univariate logistic regression of the child-level data revealed an odds ratio of 2.72 (p = 0.012) for presence of permanent tooth caries experience. Controlling for gender, grade, race/ethnicity, and language spoken at home, the odds ratio increased slightly to 2.76 (p = 0.012). CONCLUSIONS: There is a relationship between carious lesion experience in 1st grade and carious lesion incidence in the first permanent molars by 4th grade. Using the dmfs + DMFS > 0 criterion may help public health providers determine which children should receive sealants when resources do not allow the delivery of sealants to all children. PMID- 16468462 TI - Dental caries in twelve- and fifteen-year-olds: results from the basic oral health survey in Haiti. AB - OBJECTIVE: Reports on oral health investigations in Haiti are sparse. There are only three peer-reviewed published articles on oral health in Haiti. In order to construct a national dataset useful for public health planning, a representative dental caries survey of Haitian school attending children was conducted in 1999. METHODS: This survey was conducted using a modified version of the World Health Organization (WHO) Basic Oral Health Survey (BOHS) pathfinder method. Children ages 12 and 15 years old attending public or private schools in both rural and urban regions were targeted in seven of the nine geographic Departments of Haiti: each Department constituted a strata that was further stratified into the major urban center and one or more rural towns. Four trained examiners, calibrated to WHO caries criteria, conducted the survey RESULTS: Of the total 1,218 examined 12 and 15-year-olds, 31% of the 12-year-olds and 46% of the 15-year-olds had a DMFS of 1 or more, i.e., these percentages are the simple prevalence levels of dental caries for those age groups in Haiti. Mean DMFS scores were 1.01 (SE 0.09) and 2.52 (SE 0.02) for the 12- and 15-year-olds, respectively. No difference was observed between gender, while differences were found by geographical classification. Less than 1% of the children had any dental restorations. CONCLUSIONS: The findings from this investigation suggest that at the population level, permanent dentition caries in early adolescence is a minimal health problem, relative to dental caries in other neighboring Caribbean countries, as well as to other health conditions in Haiti. However, at the individual level, those children afflicted with decay are without dental services for all practical purposes. PMID- 16468463 TI - Influence of sociodemographic variables on dental service utilization and oral health among the children included in the year 2001 Spanish National Health Survey. AB - OBJECTIVES: To ascertain the possible influence of sociodemographic variables on dental service utilization and oral health among Spanish children. METHODS: This is a descriptive cross-sectional study using secondary individualized data drawn from 4,023 interviews conducted with parents or guardians of children aged 3-15 years as part of the 2001 Spanish National Health Survey. The dependent variables analyzed were use of dental services in the preceding 12 months, and caries ever. Classification of children was affirmative if their parents answered that the last time their child visited the dentist it was for a filling and/or had fillings in his/ her teeth/molars. Independent variables were sex, age, size of town or city, parents' educational level and monthly income of the family unit. RESULTS: A total of 47.1% of the study children had visited a dentist, stomatologist or dental hygienist in the preceding 12 months. Children having parents or guardians with the lowest educational level were 1.36 times (95% Cl: 1.10-1.68) more likely to have received no dental care than those having parents or guardians with the highest educational level. The likelihood of not having made use of such dental services rose 2.03-fold (95% CI: 1.58-2.61) in cases where monthly income was less than 900 euros versus greater than 1800 euros. Reported prevalence of caries for the sample as a whole was 31.45%. Children whose parents were in the lowest educational range were 1.37 times (95% CI: 1.08 1.71) more likely to have caries than those in the highest range. CONCLUSION: There is social inequality in the use of dental services and oral health among Spanish children. PMID- 16468464 TI - Florida adults' oral cancer knowledge and examination experiences. AB - OBJECTIVE: This study assessed awareness of oral cancer, knowledge of its major risk factors and clinical signs, and oral cancer examination experiences among Florida adults aged 40 years and older. METHODS: A statewide random digit dial, computer assisted telephone survey was conducted in 2002. Data from 1,773 respondents were weighted to permit statewide estimates. Bivariate analyses were used to examine awareness and knowledge of oral cancer. Multiple logistic regression analysis was used to model past-year oral cancer examination experiences of Florida's adults. RESULTS: In Florida, 15.5% of adults aged 40 years and older had never heard of oral cancer and another 40.3% reportedly knew little or nothing about it. About one-half of adults did not think oral white or red patches or bleeding could indicate oral cancer and 27.6% correctly identified three of oral cancer's major risk factors. After hearing an oral cancer exam described, just 19.5% of adults reported receiving one within the preceding 12 months. Blacks and Hispanics were significantly less likely than non-Hispanic whites to have received a recent oral cancer examination. Persons with low levels of education, those who lacked a regular dentist or source of preventive medical care, and adults who knew few or none of the clinical signs of oral cancer also were less likely to have received a recent oral cancer exam. CONCLUSIONS: There is widespread lack of awareness and knowledge in Florida regarding oral cancer and low levels of reported examination, particularly among groups experiencing disproportionately high incidence and late stage diagnosis. Increasing awareness of this disease and promoting primary and secondary prevention may help lessen the disease burden in Florida and reduce racial disparities in its outcomes. PMID- 16468465 TI - The strength of two indicators of social position on oral health among persons over the age of 80 years. AB - OBJECTIVE: The objective of the present study is to analyze how two dimensions of social position, education and social class, are associated with oral health among generally healthy, community-dwelling persons over the age of 80 years. METHODS: The present investigation is based on a sample of 157 community-dwelling individuals from The Kungsholmen Elders Oral Health Study (KEOHS) and included data from interviews and oral examinations. Social position was measured by education and social class. Oral health was measured by active coronal caries, active root caries, edentulism and use of dental services. RESULTS: The primary findings of the adjusted multivariate logistic regression analysis were that, compared to persons who had been in higher positions, persons who had been blue collar/ white-collar workers had significantly greater odds of having coronal caries and high, but nonsignificant odds of being edentulous. Further, persons with elementary/ medium education tended to forego regular dental services more than persons with high education. CONCLUSION: The study identified social inequalities in oral health even in a population of independently living, generally healthy very old Swedes and in a country where the public health policies have tried to minimize these inequalities. PMID- 16468466 TI - Toothbrushing patterns over time in at-risk metropolitan African-American 5th- 8th graders. AB - OBJECTIVES: A large study of risky pre-teen behavior provided an opportunity to examine self-reported toothbrushing frequency for stability over time and adequacy. METHODS: 1115 metropolitan African-American children at risk for violence and drug use self-reported toothbrushing frequency in at least one of five measurement points from 5th to 8th grade as part of a larger study. Longitudinal data were available for 815 students. RESULTS: 81% reported mainly twice daily, 8% reported mainly once daily, 10% changed over time, and 1% were consistently less than once daily. CONCLUSIONS: Overall, the children reported once or twice daily toothbrushing frequency, stable between 5th and 8th grades. A minority of children showed low or inconsistent frequencies and these results may indicate an opportunity for intervention to improve habits. PMID- 16468467 TI - Using tablet PC's in dental practice research: technology, cost savings, and direct data entry "on the go". AB - Recent technological developments have greatly simplified data collection, recording, and transfer in studies relying on direct observation or survey methodology, reducing both cost of data collection and the time necessary to prepare data for analysis. The purpose of this communication is to describe how tablet PC computers can be used in data collection, thereby eliminating intermediate data collection steps and chances for error, and thus reducing overall cost. The data collection procedure used to illustrate this technology is a direct observation study of the factors associated with the delivery of preventive services by dentists and hygienists in their daily practice. PMID- 16468468 TI - Commentary: optimal adjuvant endocrine therapy of postmenopausal breast cancer. AB - There is a growing body of evidence that the optimal adjuvant endocrine therapy for hormone-sensitive breast cancer in postmenopausal women should include an aromatase inhibitor. However, further research is required to establish the optimal aromatase inhibitor and whether such a drug should be used as monotherapy or in sequence after 2 to 3 years of tamoxifen. PMID- 16468469 TI - Screening and diagnostic mammograms: why the gold standard does not shine more brightly. AB - The objective of this article was to evaluate the sensitivity, specificity, positive predictive value, and negative predictive value of both screening and diagnostic mammograms. We looked at twenty seven studies found online with keywords: mammography, positive predictive value, negative predictive value, sensitivity, and specificity using the search engines google.com and botbot.com and placed them into three tables, sorting them first by purpose: either screening or diagnostic studies, and then by study size. We found a wide range of values in the studies reported and a high rate of false positives in many of them as well. Although many clinicians use the mammogram so often and rely on those results, many would benefit by being able to see the wide range of data that is reported worldwide in a format as shown in this article. PMID- 16468470 TI - The potential role of breast ductoscopy in breast cancer screening. AB - Breast cancer remains the most common malignancy among women in the Western world. Mammography, which is currently the main screening modality for early detection, has a low positive predictive value of only 25%, especially in young women with very dense breasts. Therefore, new screening approaches are needed for the early detection of breast cancer in all age groups. Mammary ductoscopy (MD) is a newly developed endoscopic technique that allows direct visualization and biopsy of the mammary ductal epithelium where most cancers originate. The procedure can be performed under local anesthesia in the office setting. At present, MD is used as a diagnostic adjunct in patients with pathological nipple discharge and to guide duct excision surgery. This article focuses on the potential of this technique in breast cancer screening and highlights its limitations in this context. PMID- 16468471 TI - The evolving role of proteomics in the early detection of breast cancer. AB - There has been emerging interest in the examination of tumor protein expression (proteomics) as a means to identify novel diagnostic and therapeutic targets in women with breast cancer. Specifically, several investigators have examined biological fluids (serum and breast ductal fluid) and breast tissue in an attempt to detect novel proteomic profiles in women with breast carcinoma. The current tools of proteomic research are evolving, but include two-dimensional polyacrylamide gel electrophoresis and mass spectrometry. Initial studies have identified several unique biomarkers and proteomic profiles that were able to discriminate between non-cancer and breast cancer patients. In the future, the application of large-scale proteomic technology may provide a means of early detection, surveillance, and identification of potential therapeutic targets. PMID- 16468472 TI - New approaches in breast cancer management: sentinel node biopsy and intraoperative radiotherapy. AB - Breast cancer is presenting earlier, and treatment is becoming less invasive. We review two recent changes in the approach to management. Sentinel lymph node biopsy is a minimally invasive technique to identify the first draining lymph node in direct communication with the primary tumor; it enables "selective lymphadenectomy." Axillary lymph node dissection is reserved as a therapeutic procedure only for proven node positive patients. The concept has been validated, the techniques have been optimized, and randomized controlled trials have confirmed lower morbidity without compromising regional control compared with conventional treatment. The procedure is considered by many as the standard of care for staging the axilla in early breast cancer, although several unanswered questions remain. Adequate training and experience in the technique are vital to ensure high sentinel node identification and low false negative rates. Intraoperative radiotherapy is an attractive concept that enables delivery of single fraction radiotherapy in the operating room immediately after resection of the primary tumor. It is convenient for patients and appears effective in pilot studies. Partial breast irradiation to the index quadrant has been practiced for many years in the form o brachytherapy. Trials are under way comparing intraoperative radiotherapy with conventional external bea irradiation. Intra operative radiotherapy should not be used outside of clinical trials until the results of the current randomized trials are known. PMID- 16468473 TI - Current status of taxanes as adjuvant therapy for early-stage breast cancer. AB - Several factors explain the significant improvement in outcome among patients diagnosed with breast cancer over the last 20 years. Chief among them are the greater adoption of screening strategies and an improved understanding of the benefit offered by adjuvant systemic therapy in reducing the risk of cancer recurrence and death. Predictive markers of response like the estrogen receptor (ER) and the human epidermal growth factor receptor 2 (HER2) help identify those most likely to derive a survival benefit from adjuvant therapy with anti estrogens and trastuzumab therapy, respectively. However, despite the lack of robust predictive markers of clinical benefit, adjuvant systemic chemotherapy is an effective tool to improve the survival odds of breast cancer patients, especially those with ER-negative breast cancer. The Oxford Overview confirmed the incremental benefit offered by the use of anthracycline-containing regimens in the adjuvant setting. After being approved in the early 1990's for the treatment of advanced breast cancer, the taxanes gradually established themselves as useful drugs in the adjuvant setting. In this commentary, we review much of the current data that support the use of taxanes in early-stage breast cancer. PMID- 16468474 TI - Exercise for breast cancer survival: the effect on cancer risk and cancer-related fatigue (CRF). AB - To date, all epidemiological research in this area has focused on the relationship between physical activity level and the risk of breast cancer in healthy women, or more recently, those who have recovered from the disease. Most of this research highlights the fact that those women who are physically active are at a reduced risk of the disease. Although physical activity is similar to exercise, it lacks the specificity of a prescribed exercise training program. Consequently, such research can only be viewed as a promising indicator of the beneficial effect that regular exercise may have for breast cancer survivors. Furthermore, due to the nature of such research, there has been a failure to provide specific evidence concerning the most suitable modality, duration, intensity, and frequency of training for risk reduction in breast cancer survivors. Thus, evidence aiding the correct prescription of exercise for this population has been lacking. More promising evidence is provided by randomized controlled trials, which examine the effect of exercise on specific risk factors and provide convincing scientific rationale for the use of exercise among breast cancer survivors. These studies not only provide understanding of the physiological mechanisms by which exercise can be effective at aiding a reduction in breast cancer risk, but also allow conclusions on the correct prescription to be drawn. Additionally, exercise has proven to be effective in combating cancer related fatigue (CRF), significantly improving both quality of life outcomes (QOL) and physiological capacity in women who have survived breast cancer. In order to promote a wider understanding of the beneficial effect that exercise holds for this population regarding reduction of breast cancer risk and CRF, this review discusses this research, making conclusions regarding the necessary training prescription to elicit such benefits. PMID- 16468475 TI - Life on the fast track. PMID- 16468477 TI - Cranial strains and malocclusion: III. Inferior vertical strain. AB - This article describes the Inferior Vertical Cranial Strain, one of the seven possible cranial strains that are discussed in this series of articles. Clinicians have to understand cranial strains to better treat their patients. There is a major link between the malocclusion we see and the underlying physiology of the patients. With airway restriction, it is necessary to understand the cranial, postural and facial factors as well as the soft tissue contribution for a more effective overall treatment of the patient. PMID- 16468476 TI - Common factors among the major malocclusions. AB - Dental practitioners tend to classify malocclusions in a schematic fashion often forgetting the full possibilities of characteristics related to a particular situation or malocclusion. This paper is designed to clarify and describe the different malocclusions, stressing the multiple faces of Class I in the mixed dentition based on the previous work of Drs. Anderson, Dewey and Sim. The description of all the major malocclusions is included with their facial, skeletal, dental and functional characteristics. Guidelines for diagnosis and treatment planning with a summary of possible difficulties in treatment are also discussed. PMID- 16468478 TI - Brainstorming. PMID- 16468479 TI - Multi-disciplined approach to orthodontics. PMID- 16468481 TI - The sandwich technique. PMID- 16468480 TI - Skeletal maturation evaluation for treatment timing in mandibular therapy. PMID- 16468482 TI - [The role of preprosthetic orthodontics in the interdisciplinary management of congenitally missing maxillary lateral incisors: case report]. AB - The early loss of permanent teeth following trauma or congenital aplasia and agenesia need to be corrected by orthodontic or prosthetic means, sometimes combined with implant therapy. Preprosthetic orthodontic measures are often an integral part of comprehensive oral rehabilitation. The individual aspects of treatment are aimed at optimizing dentofacial esthetics and at improving masticatory function and hygiene potential of prosthetic restorations. The orthodontic solution results usually in different anatomic, functional, and esthetic problems. In the present study the treatment possibilities of two patients with missing upper lateral incisors are presented. In the first case the problem of the missing upper lateral incisors was solved by adhesive bridges, after the fixed appliance orthodontic treatment, where the diasthema medianum was closed, and the upper canines were distalized. In our second case the same problem was solved by inserting implants, with single, metal free crowns in the upper lateral incisor area, after the fixed appliance orthodontic treatment, where the upper canines were distalized, and the bicuspids derotated. Prosthetic and implant procedures are very demanding and require long-term maintenance. Preprosthetic orthodontics will continue to gain significance in future esthetically-functionally oriented dentistry and that its integration into multidisciplinary rehabilitation is often indispensable. PMID- 16468484 TI - [Necrotizing sialometaplasia]. AB - Necrotizing sialometaplasia is a non-neoplastic self-limiting inflammatory ulcerous condition of the minor salivary glands. It is reported to appear in the oral cavity at every area where minor salivary glands exist, particularly those of the palate. Necrotizing sialometaplasia is a very rare disease that spontaneously develops with fearsome features. It is thought to be due to infarction of minor salivary glands, and heals in a few weeks, without any treatment. The clinical and histopathological features of necrotizing sialometaplasia often simulate those of such malignancies as squamosus cell carcinoma or mucoepidermoid carcinoma. Familiarity with necrotizing sialometaplasia and correct differential diagnosis are paramount in avoiding misdiagnosis and inappropriate treatment. PMID- 16468483 TI - [Evaluation of clinical effects of amine fluoride/stannous fluoride containing toothpaste and mouthrinse]. AB - The aim of the study was to evaluate the effects of amine fluoride (AmF)/stannous fluoride (SnF2) containing toothpaste and mouthrinse on gingival health of young adults. Altogether 42 persons participated in the study (mean age: 28.3 +/- 7.2) and were evaluated for Plaque Index (PI) (Silness and Loe, 1963) and Gingival Index (GI) (Loe and Silness, 1964). The probands were randomly divided into two groups, both of them used AmF/SnF2 containing toothpaste twice a day for three minutes toothbrushing. One group (combined group) after toothbrushing rinsed with mouthrinse containing AmF/AnF2 combination. After four weeks the participants were re-examined. At the end of the study statistically significant decrease was found in dental plaque (PI) and gingival index (GI) values. Values of PI and GI in the group using only toothpaste were as follows (mean +/- S.D.): PI values baseline: 1.03 +/- 0.20; final: 0.60 +/- 0.13; p<0.001; GI values-baseline: 0.87 +/- 0.22; final: 0.505 +/- 0.22; p<0.01. The values of PI and GI in the combined group were as follows: PI values-baseline: 1.10 +/- 0.20; final: 0.52 +/- 0.10; p<0.001; GI values- baseline: 1.05 +/- 0.27; final: 0.50 +/- 0.17; p< +/- 0.001. The reduction of PI and GI values was significant in all groups but it was higher in the combined (toothpaste + mouthrinse) group than in the one using toothpaste only. It is concluded that regular use of AmF/AnF2 toothpaste has a beneficial effect for reduction of plaque accumulation but its effect can be enhanced with additional use of Am/SnF2 containing mouthrinse. PMID- 16468485 TI - [Lexicologic parameters of free association (coupling) about teeth of Hungarian primary school children. An initial study]. AB - Free association (coupling) of 97 Hungarian primary school children (age: 8-15 yrs, 44 male, 53 female) about their teeth was collected and analysed related to lexicologic parameters, as a pilot to establish further investigations. In some cases significant (p < or = 0.05) differences within the groups related to several topics were detected in the case of the length of the text and in the case of the distribution of etymons (root of word). Gender significantly influenced the length of the text as well. Some effect of dental fear and anxiety on the length of the text, and on the etymon's distribution may also be possible. The analysis of the most frequently used words indicated some coupling of pain and fear, and the importance of the mother in how the children see dental life events. PMID- 16468486 TI - [In vitro study of the effects of 10-30 % hydrogen peroxide solutions on the surface of human tooth enamel]. AB - The aim of the present study was to investigate the effects of hydrogen peroxide solutions (10-30%) on the enamel surface and the inner structure of human teeth. Prepared enamel samples were exposed to various concentrations of hydrogen peroxide solutions for one hour. Changes in the enamel were analyzed by two different methods. The inner structure was studied by the Ellman's reagent to detect possible changes in the amount of free thiol groups in the remaining organic phase of matured enamel. Surface alterations were imaged by atomic force microscope. Ellman's reaction revealed the presence of three groups of free thiols: one with easy, the second one with medium and the third one with difficult accessibility by hydrogen peroxide. Atomic force microscopy revealed severe alterations on the enamel surface after treatment with both low and high concentration of hydrogen peroxide. Our observations indicated that hydrogen peroxide induced alterations in the surface structure and in the inner phases of mature enamel in both low and high concentrations. PMID- 16468487 TI - [The electrochemical behavior of titanium and dental casting in modelling oral conditions]. AB - The electrochemical behavior of titanium and dental casting alloys in modelling oral conditions With applying dental implants in the oral cavity the direct contact of various metals (titanium and other casting alloys) is unavoidable. Considering the electrolyte characteristic of the saliva, the chances are given that electrochemical processes might occur under such circumstances. The aim of this study was to investigate the electrochemical behavior of titanium and other dental alloys being in direct contact with the oral environment. Nickel-chromium, cobalt-chromium, high palladium content and gold alloys as well as CP grade I titanium cast samples respectively were involved in the study. Test models were made from these metals. As an electrolyte solution (set to pH 4) artificial saliva and 0.9 % NaCl solution were used. The type and the rate of corrosion were assessed with laboratory immersion test and electrochemical polarization method. In case of polarization method a three-electrode measuring-cell was applied. Laboratory exposure tests revealed that titanium resisted corrosion; there was no sign of corrosion at all. The high palladium content alloy and gold alloy behaved similarly. The nickel-chromium and cobalt-chromium alloys showed some signs of corrosion, especially in sites with damaged surface or inhomogeneities on the surface. Results of the laboratory electrochemical experiments correlated with immersion studies graphically illustrated in the paper. PMID- 16468488 TI - A minimally invasive technique ("healing response") to treat proximal ACL injuries in skeletally immature athletes. AB - This study documents outcomes of athletically active, skeletally immature patients with proximal anterior cruciate ligament (ACL) tears treated with a non reconstructive technique to promote healing ("healing response"). Between 1992 and 1998, 13 skeletally immature athletes with proximal ACL tears underwent a healing response procedure. Patients with previous ACL injury, other concurrent ligament pathology, and/or complete mid-substance ACL tears were excluded. Average preoperative KT-1000 arthrometer manual maximum difference for all patients was 5 mm (range: 3-10 mm). Preoperatively, all patients had a 1+ or 2+ pivot shift, and all patients reported knee function as abnormal or severely abnormal. Patients were followed prospectively with clinical examinations, KT 1000 testing, and subjective questionnaires. Three (23%) patients had a re-injury 30 to 55 months after the healing response and underwent subsequent ACL reconstruction. Subjective follow-up on the remaining 10 patients at an average of 69 months (range: 26-113 months) postoperatively indicated no patients experienced pain or giving way, and all considered their knee function normal. Average Lysholm score was 96, Tegner score was 8.5 (range: 7-10), and patient satisfaction at follow-up was 9.9 (1=very dissatisfied and 10=very satisfied). Clinical examination at least one year postoperatively was performed on 7 of 10 patients at 35 months (range: 12-63 months). Five patients had a negative pivot shift and 2 had a 1+ pivot shift. KT-1000 measurements improved to 2 mm (range: 0 3 mm). In the athletically active, skeletally immature patient, the healing response procedure can restore stability and knee function, with proper patient selection. In this study group, patients were very satisfied with the procedure and returned to a high level of sports and activities. PMID- 16468489 TI - Healing of a surgically created partial detachment of the posterior cruciate ligament using marrow stimulation: an experimental study in dogs. AB - The inability of intra-articular (cruciate) ligaments to generate a substantive reparative response may preclude the primary surgical repair of such lesions. A marrow stimulation technique that provides access to a fibrin scaffold, cytokines, and mesenchymal stem cells has been shown to be successful in generating a repair response in articular cartilage. We hypothesized that a similar approach may be effective in initiating and supporting a healing response in cruciate ligament injuries. The ability to treat cruciate ligament injuries by primary repair may have significant advantages over the more extensive reconstructive procedures currently advocated. Bilateral subtotal (75% of width) lacerations were made in the proximal aspect of the posterior cruciate ligament (PCL) in 10 adult mongrel dogs. A marrow stimulation technique in which small holes were made through the femoral origin of the PCL into the marrow cavity immediately adjacent to the lesion was performed in one limb. The contralateral limb served as the non-stimulated control. Twelve weeks after surgery the extent of healing and the histological character of the repair tissue in both groups were compared. Marrow stimulation resulted in a statistically significant (P<.05) improvement in the healing of subtotal lacerations made in the PCL. The marrow stimulated repair tissue was more cellular and had a more organized extracellular matrix than the repair tissue in the non-stimulated group. Our hypothesis was confirmed. Marrow stimulation can induce a "healing response" in intra-articular (cruciate) ligaments. This technique may have clinical applications in primary surgical repair of these structures. PMID- 16468490 TI - Clinical experience with the effectiveness and tolerability of hylan G-F 20 in 1047 patients with osteoarthritis of the knee. AB - This retrospective review evaluates the effectiveness and tolerability of hylan G F 20 for relief of pain due to knee osteoarthritis in a large orthopedic practice over a 5-year period. Prospectively collected data from patients who initiated intra-articular hylan G-F 20 (3 weekly injections) for osteoarthritis knee pain treatment were analyzed. Efficacy variables included physician visual analogue scale (VAS: 100 mm), and patient rating of pain, mobility, and amount of pain medication taken after treatment. Patients (n=1047; 1489 knees) were an average age of 65.3 years, 60% female, and 71% had radiologic osteoarthritis grade IV. Mean VAS scores significantly improved with hylan G-F 20 compared to baseline at all time points (P<.0001). Most knees (62%-89%) responded positively with hylan G F 20. Pain and mobility improved and less pain medication was needed after therapy. The incidences of local adverse events were 5.2% per patient and 1.2% per injection; most local adverse events were mild or moderate, with severe local events in 0.3% of injections. Our clinical experience shows that hylan G-F 20 effectively relieves osteoarthritis knee pain (as indicated) and reduces pain medication needed for up to 6 months with a low incidence of local adverse events. PMID- 16468491 TI - Biomechanical stability of a retrotubercle opening-wedge high tibial osteotomy. AB - In recent years, opening-wedge osteotomy has gained popularity. However, the complication rate reported is high. Opening-wedge osteotomy was modified to remedy the problems seen with the conventional technique including loss of correction, delayed healing, and patella infera. This biomechanical study evaluates the response of a new opening-wedge osteotomy in a static and dynamic mode of human cadavers. Results were compared to the stability of the conventional technique. Six preserved pairs of human cadaveric knees were tested. Specimens of the same pair were randomly assigned to either the modified or conventional osteotomy. Internal fixation was used to ensure precise correction and prevent bone collapse. Each tibia was loaded on a material testing system from 0 to 700 N for 10,000 cycles to simulate immediate full weight bearing in a walking individual. Specimens were then loaded to failure to determine ultimate load and stiffness of the construct. Displacement of the articular fragment and stiffness were measured during dynamic loading. Load to failure, displacement, and stiffness were measured during static testing. The modified osteotomy provided significantly greater stiffness (1392 N/mm) and smaller loss of correction (.68 mm) than the conventional osteotomy (741 N/mm; 1.76 mm) under cyclic loading conditions (P<.05). The modified retrotubercle osteotomy provides greater stiffness than the conventional osteotomy, increasing stability by 62% and minimizing loss of correction to <1 mm. The modified osteotomy eliminates the need for bone graft and provides additional strength to allow accelerated rehabilitation. PMID- 16468492 TI - Differences between extramedullary, intramedullary, and computer-aided surgery tibial alignment techniques for total knee arthroplasty. AB - Traditionally, intramedullary and extramedullary tibial alignment techniques are used to obtain neutral alignment during total knee arthroplasty. Computer assisted techniques are available to aid in obtaining alignment intraoperatively. This study analyzed the difference in tibial alignment using intramedullary and extramedullary techniques, and compared them to the resulting tibial axis as determined by a computer navigation system. The tibial alignment standard measure was determined by a spiral computed tomography (CT) study of each lower extremity. Intramedullary techniques resulted in a 1.3 degrees +/- 1.4 degrees of varus alignment and 4 degrees +/- 2.1 degrees increase in posterior slope while the extramedullary techniques resulted in 1.5 +/-1.8 degrees of valgus alignment and 1.8 degrees +/-1.1 degrees increase in posterior slope compared to the navigation system alignment. Direct measurement of metal markers on CT-scan analysis using the navigation system resulted in <0.5 degrees difference from the navigation data. Although the traditional alignment techniques are within a few degrees, the addition of the errors in all planes gives a much higher degree of error. Direct measurement of the mechanical axis using computer-guided surgical techniques for the tibia give a greater degree of accuracy compared to traditional alignment techniques. PMID- 16468493 TI - Arthroscopic removal of a broken bone staple occurring 24 years after insertion that presented as lateral joint line pain and joint locking. PMID- 16468494 TI - Bilateral displaced "bucket handle" tears of the medial meniscus in a 16-year-old girl. PMID- 16468495 TI - Minimal-incision total knee arthroplasty. PMID- 16468496 TI - Reduced-incision total knee replacement through a mini-midvastus technique. PMID- 16468497 TI - The minimally invasive subvastus approach for primary total knee arthroplasty. PMID- 16468498 TI - A technique for minimally invasive, quadriceps-sparing total knee arthroplasty. PMID- 16468499 TI - Quadriceps-sparing total knee arthroplasty using the posterior stabilized TKA design. AB - Minimally invasive TKA is in the early stages of development. Healthy skepticism is appropriate until long-term data has been reported. Early results with the quadriceps-sparing technique are encouraging. It appears to be less painful and entail a shorter recovery time. We hope the results will remain stable as the follow-up time increases. PMID- 16468500 TI - Genotoxic activities of aniline and its metabolites and their relationship to the carcinogenicity of aniline in the spleen of rats. AB - Aniline (in the form of its hydrochloride) has been shown to induce a rather rare spectrum of tumors in the spleen of Fischer 344 rats. The dose levels necessary for this carcinogenic activity were in a range where also massive effects on the blood and non-neoplastic splenotoxicity as a consequence of methemoglobinemia were to be observed. This review aimed at clarifying if aniline itself or one of its metabolites has a genotoxic potential which would explain the occurrence of the spleen tumors in rats as a result of a primary genetic activity. The database for aniline and its metabolites is extremely heterogeneous. With validated assays it ranges from a few limited Ames tests (o- and m-hydroxyacetanilide, phenylhydroxylamine, nitrosobenzene) to a broad range of studies covering all genetic endpoints partly with several studies of the same or different test systems (aniline, p-aminophenol, p-hydroxyacetanilide). This makes a direct comparison rather difficult. In addition, a varying number of results with as yet not validated systems are available for aniline and its metabolites. Most results, especially those with validated and well performed/documented studies, did not indicate a potential of aniline to induce gene mutations. In five different mouse lymphoma tests, where colony sizing was performed only in one test, aniline was positive. If this indicates a peculiar feature of a point mutagenic potential or does represent a part of the clastogenic activity for which there is evidence in vitro as well as in vivo remains to be investigated. There is little evidence for a DNA damaging potential of aniline. The clastogenic activity in vivo is confined to dose levels, which are close to lethality essentially due to hematotoxic effects. The quantitatively most important metabolites for experimental animals as well as for humans (p-aminophenol, p hydroxyacetanilide) seem to have a potential for inducing chromosomal damage in vitro and, at relatively high dose levels, also in vivo. This could be the explanation for the clastogenic effects that have been observed after high doses/concentrations with aniline. They do not induce gene mutations and there is little evidence for a DNA damaging potential. None of these metabolites revealed a splenotoxic potential comparable to that of aniline in studies with repeated or long-term administration to rats. The genotoxicity database on those metabolites with a demonstrated and marked splenotoxic potential, i.e. phenylhydroxylamine, nitrosobenzene, is unfortunately very limited and does not allow to exclude with certainty primary genotoxic events in the development of spleen tumors. But quite a number of considerations by analogy from other investigations support the conclusion that the effects in the spleen do not develop on a primary genotoxic basis. The weight of evidences suggests that the carcinogenic effects in the spleen of rats are the endstage of a chronic high-dose damage of the blood leading to a massive overload of the spleen with iron, which causes chronic oxidative stress. This conclusion, based essentially on pathomorphological observations, and analogy considerations thereof by previous authors, is herewith reconfirmed under consideration of the more recently reported studies on the genotoxicity of aniline and its metabolites, on biochemical measurements indicating oxidative stress, and on the metabolism of aniline. It is concluded that there is no relationship between the damage to the chromosomes at high, toxic doses of aniline and its major metabolites p-aminophenol/p hydroxyacetanilide and the aniline-induced spleen tumors in the rat. PMID- 16468501 TI - Is repeated head computed tomography necessary for traumatic intracranial hemorrhage? AB - This study was performed to determine the need for repeat head computed tomography (CT) in patients with blunt traumatic intracranial hemorrhage (ICH) who were initially treated nonoperatively and to determine which factors predicted observation failure or success. A total of 1,462 patients were admitted to our level II trauma center for treatment of head injury. Seventeen per cent (255/1,462) were diagnosed with ICH on initial head CT. Craniotomy was initially performed in 15.7 per cent (40/255) of patients with ICH. Two hundred sixteen patients with ICH were initially observed. Ninety-seven per cent (179/184) of observed patients with ICH and repeat head CT never underwent a craniotomy, 2.7 per cent (5/184) of patients with ICH initially observed underwent craniotomy after repeat head CT, and four patients (80%) had deteriorating neurologic status. Multivariate analysis revealed the following significant admission risk factors were associated with a need for repeat head CT indicating the need for craniotomy: treatment with anticoagulation and/or antiplatelet medications, elevated prothrombin time (PT), and age greater than 70 years. In patients with blunt traumatic intracranial hemorrhage initially observed, there is little utility of repeated head CT in the absence of deteriorating neurologic status. The only admission risk factors for a repeat CT indicating the need for craniotomy were advanced age and coagulopathy. PMID- 16468502 TI - The impact of MRI on the treatment of DCIS. AB - Magnetic resonance imaging (MRI) has been shown to detect occult invasive breast cancers with a sensitivity of 97 per cent to 100 per cent. Mammography and ultrasonography does not accurately assess the extent of ductal carcinoma in situ (DCIS), which results in a high reoperation rate. Breast MRI can improve the surgical planning in women with DCIS, improving the adequacy of initial treatment while reducing reoperation. We examined 54 patients with predominantly DCIS (> 50%) who underwent breast MRI from January 2003 to November 2004. MRI altered the surgical management in 14 (26%) patients: unilateral changed to bilateral mastectomy (5); lumpectomy or reexcision to mastectomy (3); unilateral lumpectomy or mastectomy had additional biopsies for lesions detected by MRI in the ipsilateral or contralateral breast (6). There were 8 true-positives and 7 false positives: sensitivity 86 per cent, positive predictive value 84 per cent. MRI changed the surgical management to more appropriate therapy in 15 per cent of patients avoiding additional surgery, while 11 per cent underwent negative surgical interventions. Breast MRI is a sensitive diagnostic imaging tool in patients with DCIS. However, any suspicious finding should be biopsied before a definitive operation is planned. PMID- 16468503 TI - Full-thickness chest wall resection for recurrent breast carcinoma: an institutional review and meta-analysis. AB - Locoregional recurrence of breast cancer can occur in up to 30 per cent of patients and has often been considered to indicate a poor prognosis. We reviewed our experience with full-thickness chest wall resection for recurrent breast cancer and conducted a meta-analysis of the English literature to determine patient characteristics and outcomes. Twenty-two women with isolated chest wall recurrence of breast cancer were treated between 1970 and 2000 at our institution. We reviewed their preoperative demographics, operative management and outcome, and combined our results with seven other English language studies. A majority of women (90%) underwent a mastectomy as initial management of their breast cancer. Only 18 per cent of patients had metastatic disease at the time of chest wall resection, and 71 per cent of patients had an R0 resection. The 5-year disease-free survival at City of Hope National Medical Center (COH) was 67 per cent and was 45 per cent for the entire group of 400 patients. The 5-year overall survival was 71 per cent for the COH group and 45 per cent for the entire group. Several studies reported prognostic factors, the most common being a better prognosis in patients with a disease-free interval greater than 24 months. Full thickness chest wall resection for patients with isolated local recurrence of breast cancer can provide long-term palliation and even cure in some patients. PMID- 16468504 TI - Outcomes of surgical and sonographic assessment of breast masses in women younger than 30. AB - Assessment of breast masses in young women is challenging due to normal glandular variance. Our purpose is to define the outcomes of specialized physical exam, selective breast sonography (BUS), and biopsy in women younger than 30. Five hundred forty-two patients younger than 30 referred with a palpable breast mass were studied. Patients' mean age was 24.8. Surgeon's physical exam confirmed a dominant mass in 44 per cent of cases. Thirty-seven per cent had normal clinical exams. Median tumor size was 2.2 cm. On multivariate analysis, a mass on surgeon's clinical exam (P < 0.0001), and BUS (P = 0.0001) predicted the presence of a true mass. Fifty-three per cent of self-detected abnormalities were true masses compared to 18 per cent when detected by the primary care provider (PCP) (P < 0.001). Most common diagnoses were fibroadenoma (72%), breast cysts (4%), or fibrocystic changes (3%). Malignancy occurred in 1 per cent. In summary, breast mass is a common reason for surgical consultation. Normal glandular nodularity is often mistaken for a mass. However, a judicious approach of physical exam by a surgeon using selective BUS and image guided core biopsy provides an efficient and safe approach for diagnosis. Breast malignancy is a rare but serious cause of breast mass in young women. PMID- 16468505 TI - Adverse drug reactions during lymphatic mapping and sentinel lymph node biopsy for solid neoplasms. AB - Currently, 1 per cent isosulfan blue dye and technetium-99-labeled sulfur colloid (SC) are used in lymphatic mapping (LM). Several reports have suggested that the incidence of adverse drug reactions (ADRs) during LM is high. We report our experience with LM for solid neoplasms in order to determine the incidence and risk factors for development of ADRs. Seven hundred fifty-three patients (90% women, mean age 57) underwent LM with blue dye alone or in combination with SC from 1998 to 2004. The most common malignancy was breast cancer (83%). One hundred ten patients (14%) had injection of both mapping agents. Most patients (87%) underwent intraparenchymal injection of LM agent. Eight patients (1.1%) had an ADR during LM; none had prior exposure to LM. Of these, 7 had limited reactions (mostly blue hives) that quickly resolved. One patient (0.1%) developed anaphylaxis. The ADR incidence in patients with a sulfa allergy was not significantly different than that in patients without a sulfa allergy (3.4 vs 1%, P = 0.12). No risk factors for development of ADR were identified. Overall, the incidence of ADR during LM is low. Patients with sulfa allergies and prior exposure to LM did not demonstrate an increased incidence of ADR. Anaphylaxis, though rare, can occur during LM. PMID- 16468506 TI - Metaplastic breast cancer: clinical features and outcomes. AB - Metaplastic carcinoma of the breast, a neoplasm with both epithelial and mesenchymal elements, represents less than 1 per cent of all breast cancer. We reviewed the records of all patients diagnosed with localized metaplastic breast cancer from 1991 to 2003 at our institution. We identified 21 patients. Mean primary tumor size was 4.62 cm. Eight patients (38%) had axillary node involvement at presentation. All the tumors were high grade. Only two (10%) of the tumors were hormone receptor positive. Seventeen (81%) of the patients received adjuvant chemotherapy, and 12 (57%) of the patients received radiation. Ten (29%) patients suffered a local recurrence. With a mean follow-up of 46 months, the 5-year disease-free and overall survival was 42 per cent (95% CI: 20% to 65%) and 71 per cent (95% CI: 46% to 96%), respectively. Stage-specific overall survival was 100 per cent, 83 per cent, and 53 per cent for stages I, II, and III, respectively. By multivariate analysis, there was no impact on recurrence or survival with regard to size, age, menopausal status, nodal status, histologic subtype, adjuvant therapy, or extent of surgery. Metaplastic breast cancer is a unique neoplasm that tends to present at an advanced stage and has a propensity for local recurrence. When stratified by stage, however, survival appears similar to that of adenocarcinoma of the breast, and these tumors should be treated as such. PMID- 16468507 TI - The implication of lymph node metastasis on survival in patients with well differentiated thyroid cancer. AB - Though survival for well-differentiated thyroid cancer is very good, specific populations suffer greater recurrence and mortality. Defining these cohorts can significantly influence prognosis and extent of treatment. This study, using a large, multi-institutional database, seeks to determine how the presence of lymph node disease in patients with well-differentiated thyroid cancer affects outcome. The Surveillance, Epidemiology, and End Results (SEER) database is a large-scale sample of 14 per cent of the U.S. population. It was used to identify patients with papillary and follicular thyroid carcinomas and identify the prognostic implications of lymph node metastasis. Additional factors, including presence of metastasis, age, and tumor size, were compared using multivariate and chi2 analyses. Of 19,918 patients identified, lymph node status was known for 9,904 (49.7%). On multivariate analysis, age > 45 years, presence of distant metastasis, large tumor size, and lymph node involvement significantly predicted poor outcome. Overall survival at 14 years was 82 per cent for node negative and 79 per cent for node positive patients (P < 0.05). This study shows that the survival of patients with well-differentiated thyroid cancer is adversely affected by lymph node metastases. The optimum treatment for this cohort needs further delineation, as particular populations are at greater risk of recurrence and death. PMID- 16468508 TI - Antecolic laparoscopic Roux-en-Y gastric bypass is not associated with higher complication rates. AB - Since its introduction in 1994, laparoscopic Roux-en-Y gastric bypass (LRYGB) has rapidly gained popularity for the treatment of morbid obesity. Historically, the operation is performed in a retrocolic fashion; however antecolic LRYGB has been advocated as a safe alternative. We reviewed our experience with both techniques. From January 2003 to November 2004, the new UCLA Laparoscopic Bariatric Surgery Program performed 341 LRYGBs. In March 2004, our program transitioned from a retrocolic to an antecolic approach for all gastric bypass procedures. Institutional review board approval was obtained, and the data for all patients was collected into a prospective database. The patient characteristics for the two groups were similar. The significant differences between the two groups were average body mass index and the percentage of patients with diabetes and sleep apnea. The complication profiles for the two groups were also similar. There were significant differences between the two groups in the reoperation rate, antecolic 2.0 per cent versus retrocolic 7.8 per cent, and length of stay, antecolic 2.57 versus retrocolic 2.89 days. There were no anastomotic leaks or deaths in either group. Antecolic LRYGB is safe and may be associated with fewer complications. Only long-term weight loss results and complication rates will provide a definitive answer. PMID- 16468509 TI - Device-related reoperations after laparoscopic adjustable gastric banding. AB - Laparoscopic adjustable gastric banding (LAGB) is considered a relatively safe weight loss procedure with low morbidity. When complications occur, obstruction, erosion, and port malfunction require reoperation. We retrospectively reviewed our experience with 270 consecutive patients who underwent LAGB. Device-related reoperations were performed in 26 (10%) patients. Reoperations were related to the band in 13, to port/tubing in 11, and related to both in 2 patients. Of the 15 band-related problems, it was removed in 5 (2%): slippage (3), intra-abdominal abscess (1), and during emergent operation for bleeding duodenal ulcer (1). Revision or immediate replacement was performed in 10 (4%): slippage (5), obstruction (4), and leak from the reservoir (1). Port/tubing problems were the reason for reoperations in 13 (5%): infection (5), crack at tubing-port connection (6), and port rotation (2). Port removal for infection was followed later by port replacement (average 9 months). Overall, slippage occurred in 8 (3%), obstruction in 4 (1.5%), leak from reservoir in 7 (3%), and infection in 5 (2%) patients. Fifteen device-related problems occurred during our first 100 cases and 12 subsequently (P = 0.057). Permanent LapBand loss was only 5 per cent, leading to overall rate of 95 per cent of LapBand preservation as a restrictive device. PMID- 16468510 TI - Laparoscopic distal pancreatic resection. AB - Laparoscopic resection is not an established treatment for pancreatic tumors. Previous reports, mainly in Europe and Japan, have demonstrated the potential utility of laparoscopic distal pancreatectomy (LDP). However, few reports have been published from the United States. We instituted a pilot program to assess LDP. A total of 11 patients were included from December 2003 to December 2004. All patients were staged with preoperative endoscopic ultrasound and received vaccinations for possible splenectomy. The indications for surgery were as follows: neuroendocrine tumor (n = 7), unspecified tumor (n = 1), and cystic neoplasm (n = 3). All procedures began with diagnostic laparoscopy and intraoperative ultrasound. Three patients underwent laparoscopic enucleation of a discrete pancreatic nodule. In eight patients, LDP was attempted. One patient required conversion to an open procedure. In the other seven patients, the procedure was completed laparoscopically, two with hand-assist. The average operative time was 5 hours and 3 minutes; average length of stay was 5 days; and the splenectomy rate was 57 per cent (n = 4). There was one complication of an infected hematoma. There were no pancreatic leaks, deaths, nor readmissions. LDP with or without splenectomy is feasible and can be performed with minimum morbidity and only slightly increased operative time. PMID- 16468511 TI - Positive intraoperative cholangiography during laparoscopic cholecystectomy: is laparoscopic common bile duct exploration necessary? AB - The treatment of choledocholithiasis discovered incidentally during laparoscopic cholecystectomy is not yet standardized. Options include laparoscopic common bile duct exploration (LCBDE), postoperative endoscopic retrograde cholangiopancreatography with endoscopic sphincterotomy (ERCP-ES), and no intervention. We undertook a review of our case series to determine whether LCBDE is obligatory and which LCBDE method is unsuccessful. During the 6-year study period, 91 patients with choledocholithiasis were identified. Fifty-six patients (62%) underwent LCBDE. Thirteen (23%) of these 56 patients subsequently required ERCP. Balloon sweeping of the common bile duct failed in 10 of 21 patients (48% failure) compared to any other combination of techniques with a failure rate of 1/33 (3%; P < 0.001). Two patients did not undergo complete duct exploration because of technical problems. Thirty-five patients (38%) did not undergo LCBDE. Nine of these patients (26%) did not have ERCP-ES. None of the patients who underwent postoperative ERCP-ES required additional procedures or surgery. LCBDE can successfully treat common bile duct stones, with minimal to no morbidity, but is not mandatory for safely treating choledocholithiasis. Additionally, advanced techniques for clearing the common bile duct are more successful. Surgeons should be proficient at performing these techniques. PMID- 16468512 TI - CT angiography in penetrating neck trauma reduces the need for operative neck exploration. AB - The evaluation of penetrating neck injury has evolved dramatically from mandatory operative exploration of Zone II injuries that penetrate the platysma to selective management based on physical examination and adjunctive studies. More recently, CT angiography has emerged as an efficient, noninvasive method of evaluating penetrating neck injury. We retrospectively reviewed our experience over 10 years with the management of penetrating neck injury. One hundred thirty cases were reviewed with 34 undergoing CT angiogram (Group CTA) and 96 with no CT angiogram (Group nCTA). Group CTA had significantly fewer neck explorations, 1 (3%) versus 32 (33%), P < 0.001. Negative explorations were significantly higher in nCTA as well, with a rate of 22 per cent versus 0 in CTA (P < 0.01). The use of angiogram and esophagram was also significantly lower in CTA versus nCTA (P = 0.02 and P = 0.04). Of the 34 patients in CTA, 4 (12%) also underwent angiography and 4 (12%) received a contrast esophagram. Of the 64 patients in nCTA who did not undergo a neck exploration, 19 (29%) underwent angiography, and 17 (26%) received a contrast esophagram. The use of CT angiogram increased over time with a concomitant decrease in the rate of neck explorations. PMID- 16468513 TI - Does pelvic hematoma on admission computed tomography predict active bleeding at angiography for pelvic fracture? AB - Pelvic angiography plays an increasing role in the management of pelvic fractures (PFs). Little has been written regarding the size of pelvic hematoma on admission computed tomography (CT) and how it relates to angiography results after PF. This is a retrospective review of trauma patients with PF who underwent an admission abdominal/pelvic CT scan and pelvic angiography from 2001 to 2003. CT pelvic hematoma was measured and classified as minimal or significant based on hematoma dimensions. Presence of a contrast blush on CT scan was also documented. Thirty seven patients underwent an admission CT scan and went on to pelvic angiography. Of the 22 patients with significant pelvic hematoma, 73 per cent (n = 16) had bleeding at angiography. Fifteen patients had minimal pelvic hematoma, with 67 per cent (n = 10) showing active bleeding at angiography. In addition, five of six patients (83%) with no pelvic hematoma had active bleeding at angiography. Six patients had a blush on CT scan, with five of these (83%) having a positive angiogram. But, 22 of 31 (71%) patients with no blush on CT scan had bleeding at angiography. The absence of a pelvic hematoma or contrast blush should not alter indications for pelvic angiography, as they do not reliably exclude active pelvic bleeding. PMID- 16468514 TI - Predictors of morbidity and mortality in patients with traumatic duodenal injuries. AB - The aim of our study is to determine factors that predict morbidity and mortality in patients with traumatic duodenal injury (DI). A retrospective review from July 1996 to March 2003 identified 52 patients admitted to our trauma center (age 24.4 +/- 2.1 years, ISS = 18.8 +/- 1.76). The mortality rate for patients with duodenal injury was 15.4 per cent (n = 8). The mechanisms of injury were blunt (62%), gun shot wound (GSW) (27%), and stab wound (SW) (11%). There was no difference in mortality based on mechanism of injury. Management was primarily nonoperative [n = 30 (57%)]. Of those with perforation (n = 22), 64 per cent underwent primary repair (n = 14), 23 per cent duodenal resection (n = 5), 9 per cent duodenal exclusion (n = 2), and one patient pancreaticoduodenectomy. The method of initial surgical management was not related to patient outcome. Univariate analysis demonstrated that nonsurvivors were older, more, hypotensive in the emergency department, had a more negative initial base deficit, had a lower initial arterial pH, and had a higher Injury Severity Score. Nonsurvivors were also more likely to have an associated inferior vena cava (IVC) injury. Multivariate regression analysis revealed age, initial lowest pH, and Glasgow Coma Score to be independent predictors of mortality, suggesting that the physiologic presentation of the patient is the most important factor in predicting mortality in patients with traumatic DIs. PMID- 16468515 TI - Is field hypotension a reliable indicator of significant injury in trauma patients who are normotensive on arrival to the emergency department? AB - There is a subset of trauma patients who are hypotensive in the field but normotensive on arrival to the emergency department (ED). Our objective was to evaluate the presence, type, and severity of injuries in these patients. Data were retrospectively reviewed from patients treated at a level 1 trauma center over 1 year. Hypotension was defined as systolic blood pressure (SBP) less than 90 mm Hg. Forty-seven patients were included. The mechanism of injury was blunt in 37 patients and penetrating in 10. The average field SBP was 76 +/- 11 mm Hg. The average SBP on arrival to the ED was 120 +/- 19 mm Hg. The average injury severity score (ISS) was 16.3 +/- 10.3 (range, 1-43). Twenty-four patients (51%) had significant injury (ISS > or = 16). Nine patients (19%) had critical injury (ISS > or = 25). Twenty-six patients (55%) required surgery, and 43 (91%) required ICU admission. Common injury sites included the head and neck (57%), thorax (44%), pelvis and extremities (40%), and abdomen (34%). Overall mortality was 10 per cent (n = 5). All patients that died had significant head and neck injuries (AIS > or = 3). Field hypotension was a significant marker for potential serious internal injury requiring prompt diagnostic workup. PMID- 16468516 TI - The quality of surgical resident interpretation of after-hour CT scans of the head and abdomen in trauma. AB - Computed tomography (CT) is an important diagnostic tool in the evaluation of trauma patients. Accurate interpretation of CT scans remains critical in decision making and the resultant quality of care. In our study, the records of a consecutive cohort of trauma patients who underwent after-hour CT scans of the head and abdomen between January 23 and June 30, 2004, at Kern Medical Center were reviewed. Three hundred thirty-five CT studies were collected in 211 patients. The accuracy of resident interpretation was 92.8 per cent for abdominal and 97.5 per cent for head CT. Resident readings were 93.2 per cent sensitive and 95.4 per cent specific with a positive predictive value of 85.2 per cent and a negative predictive value of 98.0 per cent. There were 16 (4.8%) instances of interpretation discrepancy between the surgical resident and attending radiologist. Most differences occurred in the evaluation of abdominal CT. In no instance was management or outcome for these patients affected. This data demonstrates a low error rate in resident interpretation of after-hour CT scans of the head and abdomen in trauma, but there remains a need for the continued review of the quality of surgical resident radiologic interpretation in situations when an attending radiologist is not immediately available. PMID- 16468517 TI - Use of recombinant factor VIIa for adjunctive hemorrhage control in trauma and surgical patients. AB - Recombinant factor VIIa (rFVIIa) has recently been described for patients with ongoing massive bleeding in a number of different clinical scenarios. A retrospective chart review was conducted at a public level I trauma center in order to describe the use of rFVIIa in trauma and surgical patients with massive bleeding despite surgical control. Fifteen trauma and general surgical patients underwent major operative procedures and developed coagulopathy requiring massive blood product transfusion. All patients had continued life-threatening hemorrhage despite surgical control of bleeding. The mean base deficit was 6 and arterial lactate was 9.0 mmol/L. An initial dose of rFVIIa was given intravenously, followed by a second dose if there was evidence of at least a partial response. Twelve of 15 patients who had been expected to die from hemorrhage survived for greater than 48 hours, and 7 survived to hospital discharge. A partial or complete hemostatic response to rFVIIa was noted in 12 of 15 patients. The number of blood products received after administration of rFVIIa was significantly reduced and the International Normalized Ratio (INR) decreased. Our experience demonstrates that rFVIIa may reduce or completely arrest coagulopathic bleeding in trauma and surgical patients after vascular control. PMID- 16468518 TI - The effect of older blood on mortality, need for ICU care, and the length of ICU stay after major trauma. AB - The purpose of this study was to determine if the quantity and age of blood is an independent risk factor for in-hospital mortality, need for intensive care unit (ICU) care, and an increased length of stay in the ICU. This was a retrospective cohort study performed at a level I trauma center between 2001 and 2003. Consecutive trauma patients who received at least 1 unit of packed red blood cells (PRBCs) were included. The number of units of PRBCs transfused and the ages of each unit of PRBCs were recorded. Other variables including the patient's age, sex, Trauma-Related Injury Severity Score (TRISS), and whether the blood was leukopoor were collected. End points included in-hospital mortality, need for ICU care, and the length of stay in the ICU (in days). Multivariable logistic and Poisson regression analyses were performed to model the independent effect of the dose of aged blood (defined as the product of the average age of all units received and the total number of units received) with respect to each end point while controlling for age, TRISS, the total number of units administered, and the proportion of blood that was leukopoor. During the study period, 275 patients were studied. Patients who received older blood had a significantly longer ICU stay (RR 1.15, 95% CI: 1.11-1.20), possibly reflecting a higher level of organ dysfunction. Patients who received older blood, however, did not have a significantly higher in-hospital mortality rate (OR 1.21, 95% CI: 0.87-1.69) or a significantly higher need for ICU care (OR 1.20, 95% CI: 0.87-1.64). The quantity of aged blood is an independent risk factor for length of ICU care. This may be a proxy indicator for multiple organ failure. Further research is required to define which patients may benefit from newer blood. PMID- 16468519 TI - Impact of an autotransfusion system during endoluminal repair of abdominal aortic aneurysms: does it decrease the use of allogeneic blood? AB - Endoluminal repair of abdominal aortic aneurysms (AAA) is associated with decreased blood loss and transfusion requirements when compared to open AAA repair. We evaluated the use of an intraoperative autotransfusion system during endoluminal exclusion of AAAs. Fifty endoluminal AAA procedures selected at random were reviewed retrospectively. Patients were divided into 4 groups according to their estimated blood loss: Group I, 20 patients (0-500 cc); Group II, 18 patients (501-1,000 cc); Group III, 7 patients (1,001-1,500 cc); and Group IV, 5 patients (1,501-3,400 cc). The average blood loss was 327 cc for Group I, 728 cc for Group II, 1,217 for Group III, and 2,125, for Group IV. The overall blood loss was 834 cc, but 75 per cent was recovered. Hematuria was always present when greater than 1,000 cc of blood was retransfused, but renal function was not affected even with the concomitant administration of radiographic contrast. Only one patient required an autologous blood transfusion. The use of an autotransfusion device is safe, efficacious, and appears to further limit the need for homologous blood transfusion in patients undergoing endoluminal AAA repair. PMID- 16468520 TI - VATS anatomic pulmonary resection in octogenarians. AB - Although modern techniques in anesthesia and surgery have reduced morbidity and mortality for pulmonary resection, some physicians still consider advanced age a contraindication to resection of lung cancer. We examined our experience with VATS lobectomy in octogenarians at Cedars-Sinai Medical Center over 12 years (1992-2004). There were 159 patients. Mean age was 83 years (range, 80-94 years) consisting of 61 males (38%) and 96 females (62%). Operations included 153 lobectomies (96%), 3 bilobectomies (2%), and 3 pneumonectomies (2%). Two operations were converted to thoracotomy (1%), one due to bleeding, and one due to poor visualization. Median hospital stay was 4.00 +/- 6.39 days. One hundred thirty-one patients (82%) had no complications. The most common complication was arrhythmias occurring in 8/159 (5%) patients. There were three perioperative deaths (1.8%). Pathology revealed 104 adenocarcinomas (65%), 25 squamous cell carcinomas (16%), 5 adeno-squamous carcinomas (3%), 7 bronchoalveolar carcinomas (4%), 7 large cell carcinomas (4%), 4 carcinoid tumors (3%), 4 non-small cell lung cancer (NSCLC) (3%), 1 mucoepidermoid carcinoma (< 1%), 1 lymphoma (< 1%), and 1 pulmonary metastasis (< 1%). Median follow-up was 29 months. The results of this series show that age alone is not a contraindication to the surgical treatment of lung cancer. PMID- 16468521 TI - Administration of nesiritide in patients after coronary artery bypass surgery induces brisk diuresis. AB - Recombinant human brain natriuretic peptide, nesiritide, has recently been used in limited studies to enhance postoperative diuresis. A retrospective chart review was conducted at a university hospital to assess the efficacy of nesiritide in cardiac surgery patients with fluid overload refractory to diuretics and dopamine. Nine out of 137 patients who underwent coronary artery bypass grafting at the institution from May 2003 to July 2004 exhibited fluid overload despite diuretics and dopamine. Those who did not respond to the therapy, as manifested by oliguria and heart failure, were started on nesiritide. Urine output, weight change, central venous pressure (CVP), pulmonary artery wedge pressure (PAWP), and serum creatinine were the main outcome measures. Within 6 hours after initiation of nesiritide, the average urine output increased from 28 to 130 mL/h. Serum creatinine levels were not significantly different after 24 hours. The mean CVP decreased from 14 to 10 within 12 hours while the PAWP decreased from 24 to 17 mm Hg. Systemic pressures did not change. One patient had to eventually undergo hemodialysis for complications of renal failure. Our experience demonstrates that infusion of nesiritide in patients with heart failure and fluid overload improves diuresis and hemodynamics without major side effects. PMID- 16468523 TI - Negative appendectomy rate: influence of CT scans. AB - Negative appendectomy rate varies significantly depending on patient age and sex. However, the impact of computed tomography (CT) scans on the diagnosis of appendicitis is unknown. The goal of this study was to examine the negative appendectomy rate using a statewide database and analyze the association of receipt of CT scan. Using the California Inpatient File, all patients undergoing appendectomy in 1999-2000 were identified (n = 75,452). Demographic and clinical data were analyzed, including procedure approach (open vs laparoscopic) and appendicitis type (negative, simple, abscess, peritonitis). Patients with CT scans performed were identified to compare the negative appendectomy rate. For the entire cohort, appendicitis type was 59 per cent simple, 10 per cent with abscess, 18.7 per cent with peritonitis, and 9.3 per cent negative. Males had a lower rate of negative appendicitis than females (6.0% vs 13.4%, P < 0.0001). The use of CT scans was associated with an overall lower negative appendectomy rate for females, especially in the < 5 years and > 45 years age categories. Use of CT scans in males does not appear to be efficacious, as the negative appendectomy rates were similar across all age categories. In conclusion, use of CT was associated with lower rate of negative appendectomy, depending on patient age and sex. PMID- 16468522 TI - Emerging trends in peptic ulcer disease and damage control surgery in the H. pylori era. AB - The prevalence of peptic ulcer disease (PUD) and the frequency of operation have been decreasing for decades. Immigration of patients harboring Helicobacter pylori may reverse these longstanding declines. The experience with a large public hospital population in an area of high immigration may portend future national trends. A 10-year retrospective study analyzed the changing demographics of PUD and the frequency and nature of surgical intervention. A total of 2,182 patients were diagnosed with PUD, 1,173 in the early period (1995-1999) and 1,009 in the recent period (2000-2004). The proportion of Hispanic patients increased from 39.3 per cent to 47.5 per cent (P = 0.017). The ratio of male to female patients decreased from 1.7:1 to 1.3:1 (P = 0.003). The PUD operation rate decreased from 6.7 per cent to 3.8 per cent (P = 0.004). Among operated patients, the frequency of H. pylori testing increased from 41.8 per cent to 81.6 per cent (P = 0.039). Acute perforation and bleeding necessitated the vast majority (87.2%) of operations. The use of acid-reducing operations declined from 50.6 per cent to 31.6 per cent in favor of nonacid-reducing "damage control" procedures. Contrary to historic trends, in the predominately immigrant public hospital patient population studied, 1) the incidence of PUD is decreasing only modestly, 2) male predominance is disappearing, 3) gastric ulcer (GU) is more prevalent than duodenal ulcer (DU), but DU requires operation more frequently than GU, and 4) there is a marked decrease in use of acid-reducing operations reflecting a new "damage control" surgical approach to acute PUD complications in the H. pylori era. PMID- 16468524 TI - Appendectomy during early pregnancy: what is the preferred surgical approach? AB - Appendectomy is the most common nongynecologic surgery performed during pregnancy. Although many surgeons offer laparoscopic appendectomy (LA) as an alternative to open appendectomy (OA) during early pregnancy, few studies have compared the effects of LA versus OA on the fetus and the outcome of the pregnancy. Twenty-eight consecutive females undergoing appendectomy for presumed appendicitis in the first two trimesters of pregnancy between January 2000 and December 2002 were identified retrospectively. Demographic information, weeks of pregnancy at operation, and surgical approach (LA or OA) were recorded. Study outcomes included operative and pregnancy-related complications, length of hospitalization, final outcome of pregnancy, and infant birth weight. Seventeen LA and 11 OA were performed. There were no significant differences in surgical or obstetrical complications, length of hospitalization, or birth weight between the two groups. Two cases of postoperative fetal demise were noted in the LA group. Although not statistically significant, the two fetal losses in the laparoscopic group are concerning. The current study did not demonstrate any advantages to the laparoscopic approach. Pending further investigation, the open approach may be preferred for appendectomy in pregnant patients during the first two trimesters of pregnancy. PMID- 16468525 TI - Laparoscopic versus open appendectomy: is the postoperative infectious complication rate different? AB - Despite the reported advantages of laparoscopic appendectomy (LA), there is debate about the postoperative infectious complication rate. Our study attempts to determine if the infectious complication rate between LA and open appendectomy (OA) is different. A retrospective review was conducted of all patients who underwent appendectomy at Kern Medical Center between 1999 and 2003. Age, sex, white blood cell count, temperature, pathology, and postoperative complications were identified. Fifty-seven patients underwent LA, and 159 patients underwent OA. The groups were well matched for demographics, white blood cell count, temperature, and percent perforated appendicitis. There was an overall 9.3 per cent complication rate. The infectious complication rate in OA versus LA group was statistically different (6.3% vs 17.6%, P = 0.04). The infectious complication rate in the LA group was significantly higher than in the OA group. Further large randomized trials are necessary to confirm our findings and to identify if LA is appropriate for a subset of appendicitis patients. PMID- 16468526 TI - Does initial choice of antimicrobial therapy affect length of stay for patients with complicated intra-abdominal infections? AB - Outcomes for complicated intra-abdominal infection are influenced by operation for source control, patient-related factors, and medical management, including antibiotic treatment. We analyzed length of stay (LOS) at 33 hospitals for 2,150 patients discharged between February 2002 and June 2003, who were > 18 years, had intra-abdominal infection, and received one of 6 first-line antimicrobials. A regression tree analysis selected important variables, their interactions, and their order of significance in explaining LOS. A linear mixed model evaluated the difference in LOS between treatment groups. Adjusted LOS was calculated by the least squares means from the model and was used to assess treatment differences. Mean LOS analyzed by initial antimicrobial therapy and stratified by diagnosis showed LOS for ampicillin/sulbactam and ertapenem to be significantly shorter from levofloxacin, ceftriaxone, and piperacillin/tazobactam (all P < 0.05). Adjusting for all other factors, the variables associated with severity (e.g., diagnosis, ICU stay, and comorbidities) had the greatest impact on adjusted LOS (all P < 0.001). Our findings indicate ampicillin/sulbactam and ertapenem were associated with shorter hospital stays, which may be explained by unaccounted for underlying severity of infection and/or by surgeons stratifying antimicrobial selection according to severity of illness. PMID- 16468527 TI - Elevated platelet count before ileal pouch-anal anastomosis for ulcerative colitis is associated with the development of chronic pouchitis. AB - Acute pouchitis (AP) after ileal pouch-anal anastomosis (IPAA) is common and easily treated. However, chronic pouchitis (CP) remains a difficult management problem and may represent a form of Crohn disease (CD) of the ileal pouch. Because CD patients have higher platelet counts than ulcerative colotis (UC) patients, we prospectively evaluated the association between preoperative platelet count and pouchitis development in 159 patients undergoing IPAA. Reactive thrombocytosis (RT) was defined as a platelet count > 450 x 10(9)/L. Median preoperative platelet count was 312 x 10(9)/L (range, 103 x 10(9)/L to 886 x 10(9)/L). One hundred twenty-five patients (79%) had a normal (150 x 10(9)/L to 450 x 10(9)/L) platelet count (-RT patient group). Twenty-eight patients (18%) had RT. Six patients (3%) had a platelet count below 150 x 10(9)/L. After a median follow-up of 13 months, 45 patients (28%) developed pouchitis. Pouchitis developed in 33 +RT patients (26%) versus 9 -RT patients (32%) (P = NS). UC patients who had +RT had a 25 per cent incidence of CP compared to only 7 per cent of those UC patients who had -RT (P = 0.03). The incidence of CP was significantly higher after IPAA in UC patients having thrombocytosis before surgery compared to UC patients having a normal platelet count before surgery. PMID- 16468529 TI - Weekly reading assignments and examinations result in sustained improvement in American Board of Surgery In-Training Examination (ABSITE) scores. AB - The American Board of Surgery In-Training Examination (ABSITE) score is used by general surgery training programs as a measure of the medical knowledge component of the Accreditation Council of Graduate Medical Education competencies. Poor performance on ABSITE (< 35%) has correlated with high failure rates on the American Board of Surgery Qualifying Examination. We previously demonstrated an improvement in ABSITE scores over a 1-year period through the initiation of weekly reading assignments and review examinations. We sought to determine whether this educational endeavor would result in a sustained improvement in ABSITE scores over several years. ABSITE scores from two successive, 3-year time periods (1999-2001, Group 1, 2002-2004, Group 2), were compared. For Group 1, no formal reading assignments or preparatory exams were given. For Group 2, weekly reading assignments and examinations were administered by the faculty. For Group 1, the mean ABSITE score was 60 per cent, versus 75 per cent (P < 0.01) for Group 2. In Group 1, 13 out of 76 scores (17%) were below 35 per cent, whereas in Group 2, only 4 out of 74 scores (5%) were below 35 per cent (P = 0.03). Sustained improvement in ABSITE scores and fewer scores < 35 per cent were achieved over a 3-year period with an educational program of weekly topic-specific reading assignments and written review examinations. PMID- 16468528 TI - Preoperative oral rofecoxib and postoperative pain in patients after laparoscopic cholecystectomy: a prospective, randomized, double-blinded, placebo-controlled trial. AB - Cyclooxygenase-2 (COX-2) inhibitors are a class of drugs that may avoid some of the side effects of narcotics and nonsteroidal anti-inflammatory drugs (NSAIDs). We performed a randomized, double-blinded, placebo-controlled trial giving a single oral dose of the COX-2 inhibitor rofecoxib 25 mg or placebo preoperatively to determine the impact upon postoperative pain, complications, narcotic use, and hospital stay after laparoscopic cholecystectomy. Investigators and patients were blinded. Pain was measured on a 10-point visual analogue scale. Eighty patients were randomized: 40 to the rofecoxib group and 40 to the placebo group. The amount of pain between the two groups postoperatively was equivalent. Pain was recorded at 1 hour, 4.03 +/- 1.93 in the rofecoxib group versus 4.38 +/- 1.34 in the placebo group (P = 0.36); at 6 hours, 3.00 +/- 1.12 in the rofecoxib group versus 2.78 +/- 0.78 in the placebo group (P = 0.42); and at 24 hours, 1.64 +/- 0.67 in the rofecoxib group versus 2.68 +/- 1.90 in the placebo group (P = 0.17). The amount of pain medication received and lengths of hospital stay was not significantly different between the two groups. Our data demonstrate no significant benefit of preoperative oral rofecoxib in patients undergoing laparoscopic cholecystectomy. PMID- 16468530 TI - Cardiac morbidity and mortality after surgery for gastrointestinal carcinomas. AB - The number of Americans undergoing surgery for gastrointestinal (GI) cancer is increasing, as is the prevalence of cardiovascular disease. Clinical risk factors have been found to be useful in predicting cardiac events after vascular procedures. Their utility for predicting cardiac events after GI carcinoma surgery is unclear. We performed a retrospective review in order to determine whether clinical risk factors are useful in predicting cardiac events in patients undergoing GI carcinoma surgery and to ascertain the incidence of postoperative cardiac events. From 1998 to 2003, 333 patients were identified, with an average age of 56 years. One hundred one (30.3%) patients had one or more clinical risk factors. The overall cardiac event rate was 3.9 per cent. Age > 70 years was the only risk factor associated with a cardiac event. There was a trend toward increased cardiac risk with increasing number of risk factors. In the absence of clinical risk factors, cardiac events after surgery for GI carcinoma are low. There is an increased cardiac risk in patients > 70 years and a trend toward increased cardiac events as the number of clinical risk factors increases. PMID- 16468531 TI - Contemporary indications for and early outcomes of abdominoperineal resection. AB - The purpose of this study was to review and characterize the indications and early outcomes of abdominoperineal resection (APR) when used in a colorectal practice in an academic setting. Data was collected from the charts of all patients undergoing APR in a retrospective manner. Data collected included demographic information and details regarding the clinical presentation. Operative factors, information regarding the postoperative course, and morbidity and mortality were evaluated. Forty-four patients were treated with an APR in this practice between the years 1992 and 2004. The indications for operation were primary rectal cancer (n = 31), recurrent rectal cancer (n = 6), intractable Crohn disease (n = 3), anal melanoma (n = 1), cloacogenic cancer (n = 1), squamous cell cancer (n = 1), and gastrointestinal stromal tumor (n = 1). Complications in the first 60 days affected 14 patients (32%). The most common complication was intra-abdominal/pelvic abscess formation occurring in 6 of these 14 patients (43%). Additional complications in the first 60 days included rectus flap necrosis, perineal wound evisceration, prolonged ileus, and urinary retention. There was no surgical mortality. Long-term complications occurred in 7 patients (16%), with parastomal hernia being the most common (43%). Although relatively infrequently used, APR will continue to play a role for selected patients in the future. Despite the significant morbidity associated with this surgery, APR may provide beneficial treatment for select cases of low rectal cancer, end-stage inflammatory bowel disease, and anal malignancies. PMID- 16468532 TI - Ultrasonography for interval assessment in the nonoperative management of hepatic trauma. AB - Abdominal ultrasonography (US) is gaining widespread acceptance as a valuable diagnostic tool in the initial evaluation of trauma victims. We investigated the utility of US as a follow-up radiologic study in nonoperative hepatic trauma. Patients with liver injury designated for nonoperative management were prospectively studied over a 2-year period at our primary adult resource center for trauma. Computed tomography (CT) and radiologist-performed US were obtained at admission and at 1 week. The ability of US to detect lesions, fluid, and complications was evaluated by comparing with the corresponding CT. Twenty-five hepatic trauma patients in the study were successfully managed nonoperatively and had both initial and follow-up US and CT scans: 1 (4%) grade I, 5 (20%) grade II, 7 (28%) grade III, 7 (28%) grade IV, and 5 (20%) grade V. Four complications developed [biloma (3) and biliary fistula (1)] in 3 patients with grade IV injury and 1 with a grade II injury. Interval US appropriately detected a complication or confirmed the absence of complication in all (13/13, 100%) patients with low grade (I-III) injury and only missed a small biloma in one patient with a grade IV injury. Interval US and CT agreement was 92 per cent for change in hemoperitoneum or parenchymal lesion. Ultrasonography is a convenient imaging modality in the evaluation of hepatic trauma. US is sufficient to detect or exclude complications in low-grade injuries. In high-grade injuries, US may be an adjunct to CT for definitive interval assessment. PMID- 16468533 TI - Recent trends in the management of combined pancreatoduodenal injuries. AB - In an effort to better characterize the natural history of pancreatoduodenal injuries, we present a review of clinical experiences in the treatment of combined traumatic pancreatoduodenal injuries, focusing on patients in extremis. Records of patients with abdominal trauma admitted to a level 1 trauma center from 1997 to 2001 were reviewed. Of 240 patients who sustained a pancreatic or duodenal injury, 33 had combined pancreatoduodenal injuries. Eighty-two per cent of the patients (27/33) in this series had penetrating injuries, 72 per cent (24) sustained gunshot wounds (GSW). Thirty-one patients were male, and the mean age was 33 years (range, 7-74). These patients presented with an average Injury Severity Score (ISS) of 22 +/- 12 and an average Glasgow Coma Score of 14 +/- 2. Overall length of stay was 39 +/- 59 days (range, 0-351 days). These 33 patients underwent a total of 57 laparotomies with an average of 1.7 operations per patient (range, 1 to 5 operations). Eighty-four per cent of the patients had an associated gastrointestinal injury and 45 per cent had a major vascular injury. Thirteen of the 33 (39%) patients presented in extremis, all 13 underwent an abbreviated laparotomy. The complication rate was 36 per cent, including fistula, abscess, pancreatitis, and organ dysfunction. There were 6 hospital deaths for a mortality rate of 18 per cent. Pancreatoduodenal injuries are associated with a variety of other serious injuries, which add to the overall complexity of these patients. Abbreviated laparotomy may be helpful when managing combined pancreatoduodenal injuries in patients who are in extremis. PMID- 16468534 TI - Extensive mobile thrombus of the internal carotid artery: a case report, treatment options, and a review of the literature. AB - The presence of a carotid stenosis, a floating thrombus, and a patient with clinical and CT evidence of a stroke represents a significant therapeutic dilemma to the clinician. The evidence of a stroke precludes any active treatment of the carotid stenosis safely, while the floating thrombus demands immediate attention. We recently were involved with just such a patient and chose a conservative approach of anticoagulation followed by operative intervention several weeks later. PMID- 16468535 TI - Portal vein thrombosis after laparoscopic colectomy: thrombolytic therapy via the superior mesenteric vein. AB - Portal vein thrombosis is a rare but well-reported complication after laparoscopic surgery. We present a case of portomesenteric venous thrombosis that occurred 8 days after a laparoscopic-assisted right hemicolectomy. Systemic anticoagulation failed to improve symptoms. The early postoperative state precluded the use of transarterial thrombolytic therapy. Transjugular intrahepatic catheter-directed infusion of urokinase into the superior mesenteric vein resulted in clearance of thrombus and resolution of symptoms. The published data on laparoscopy-induced splanchnic venous thrombosis and transjugular intrahepatic intramesenteric thrombolysis are discussed. PMID- 16468536 TI - Hyaluronic acid-carboxycellulose membrane (Seprafilm) reduces early postoperative small bowel obstruction in gastrointestinal surgery. AB - This study aimed to evaluate the efficacy of sodium hyaluronate and carboxymethylcellulose membrane (Seprafilm) on early postoperative small bowel obstruction (EPSBO). It also examined whether using Seprafilm affected surgical site infection rates in gastrointestinal surgery. One hundred eighty-four patients who had Seprafilm placed during gastrointestinal surgery between October 2000 and December 2003 were included in the study (Seprafilm group) and were compared with a matched cohort (control group) of patients operated on without Seprafilm. All patients were retrospectively assessed for EPSBO and surgical site infections. The incidence of EPSBO was significantly lower (P < 0.05) in the Seprafilm group (12/184) than in the control group (26/183). The incidence of surgical site infection between the Seprafilm (15.2%) and control (13.7%) groups was not statistically significant. The placement of Seprafilm helped to prevent EPSBO and had no significant adverse affect on surgical site infections. PMID- 16468537 TI - Long-term results of adrenalectomy in patients with aldosterone-producing adenomas: multivariate analysis of factors affecting unresolved hypertension and review of the literature. AB - The long-term surgical cure rate of patients with primary aldosteronism varies widely, and causes of persistent hypertension are not completely established. We reviewed retrospectively charts from 98 patients (range, 19-70 years old) with aldosterone-producing adenomas who underwent unilateral adrenalectomy. At a median follow-up of 81 months (range, 18-186 months), the mean blood pressure values improved in 95 out of 98 (96.9%) patients, although hypertension was cured only in 71 out of 98 (72.4%) patients. Multivariate analysis using a logistic regression model adjusted for duration of follow-up showed that only age of the patients and duration of the disease independently correlated with unresolved hypertension. The cumulative odds ratio (OR), obtained using the logistic regression function, was 5.38 (95% CI 1.78-16.22), and the OR of single variables were 1.32 (95% CI 0.36-19.83) and 4.56 (95% CI 1.41-14.78), respectively. By using discriminant analysis to derive a classification function for the prediction of unresolved hypertension, a maximum predictive power of 75 per cent was achieved. In conclusion, in patients with an aldosterone-producing adenoma undergoing surgery, the combination of age and duration of hypertension gave the best predictive power of a linear classification function and represented the main independent risk factors affecting hypertension cure rate. PMID- 16468538 TI - Crohn's disease presenting as necrotizing fasciitis: a case report. PMID- 16468539 TI - Profound refractory hypocalcemia after thyroidectomy in a patient receiving chronic oral bisphosphonate therapy. AB - The use of oral bisphosphonates in the primary care setting is increasing, most commonly to preserve bone density and prevent the development or progression of osteoporosis. We report a case of profound hypocalcemia occurring in a patient who underwent thyroidectomy while receiving chronic oral risedronate therapy for osteoporosis. In addition, the hypocalcemia was refractory to treatment for several days, necessitating a prolonged hospital stay. A review of the literature yielded no similar cases to date. We propose that the frequently observed transient hypocalcemia after thyroidectomy was exacerbated by the alterations in calcium homeostasis produced by bisphosphonate therapy and urge cessation of these agents for several weeks prior to surgery. A review of the relevant literature is provided. PMID- 16468540 TI - Hashimoto thyroiditis coexistent with papillary thyroid carcinoma. AB - Several studies report a higher rate of papillary thyroid carcinomas (PTC) in patients with Hashimoto thyroiditis (HT), indicating a possible correlation between the two diseases. We studied a group of 89 subjects undergoing surgery for thyroid carcinomas compared with a control group of 89 subjects operated on for normofunctioning goiter, and a second group of 47 patients undergoing total thyroidectomy for HT. Association with HT was found in 19 of the 71 PTC subjects (26.7%) and in 8 goiter patients (8.9%), which was a significant difference (P < 0.02). Thirteen of the HT patients, mostly with the nodular form, showed coexistent PTC (27.6%). HT and PTC coexisted in several morphological, immunohistochemical, and biomolecular aspects; increased incidence of PTC in HT patients might therefore indicate that HT is a precursor of thyroid cancer. Further studies are required, however, in order to confirm this hypothesis; until then, HT patients should undergo careful clinical and technical follow-up. PMID- 16468541 TI - Delayed diagnosis and management of an "occult" stab wound to the heart. AB - Stab wounds to the heart can present without clinical evidence of injury. Echocardiography has been advocated as an alternative to subxyphoid exploration in these cases. We present a case in which two serial echocardiography exams missed a diagnosis. Electrocardiographic changes and elevation of serial troponin I levels suggested the diagnosis, which was confirmed by thoracoscopy. Repair was facilitated by use of adenosine cardiac standstill. PMID- 16468542 TI - Successful treatment of Bouveret syndrome using holmium: YAG laser lithotripsy. AB - Although gallstone disease is highly prevalent, cholelithiasis causing gallstone ileus is uncommon. Consideration has been given for nonoperative strategies to resolve obstruction due to the significant age and comorbidities afflicting this population. A 94-year-old man presented with a 5-day history of abdominal distension and tenderness. CT scan revealed multiple large gallstones within the gallbladder, pneumobilia, and two ectopic gallstones (antrum of the stomach and distal ileum). The patient was taken to the operating room where an enterolithotomy and gastrotomy was performed with removal of gallstones and subsequent relief of obstruction. During the postoperative course, the patient developed symptoms of gastric outlet obstruction and underwent gastrointestinal endoscopy for diagnosis and treatment. Two large gallstones, present in the duodenum, were retracted into the stomach using a Roth net but could not be retrieved beyond the upper esophageal sphincter. A holmium: yttrium-aluminum garnet (Holmium: YAG) laser was used for fragmentation of the stones, with subsequent successful removal. This is the first documented successful use of the holmium: YAG laser for the treatment of recurrent gallstone ileus. Physicians should remember that in a small but important subgroup of patients, endoscopy accompanied by laser lithotripsy may prove beneficial. PMID- 16468543 TI - Families' perception of the value of timed daily "family rounds" in a trauma ICU. AB - Daily communications between the ICU trauma patients' families and the trauma team are often limited due to the unpredictable nature of subsequent patient admissions and operative procedures. In order to improve the lines of family physician communication and educate residents regarding family communication, our level I trauma center instituted daily "Family Rounds" (FR). FR occur at the same time every day, in the patient's ICU room. The purpose of this study was to determine whether families valued the scheduled daily FR, to establish whether FR improved the family-physician relationship, and to delineate strengths and weaknesses of the present structure of our FR. We mailed surveys to family members of trauma patients hospitalized in the trauma ICU for > or = 3 days. A total of 55 (22%) families responded. Combining "excellent" and "good" responses, 86.5 per cent of families looked forward to having a specific time of day to meet with the trauma team, and 90 per cent liked having rounds in the ICU room with the patient. However, 36 per cent did not like having only scheduled time for FR. The majority, 75 per cent, believed that all concerns were addressed during FR, and 84.9 per cent rated their overall experience as either excellent or good. Scheduled FR appear to improve communication between trauma surgeons and patients' families, enhance the family-physician relationship, and strengthen our surgical residency teaching program. PMID- 16468544 TI - Advances in treatment of colorectal cancer. AB - The purpose of this review is to provide the practicing surgeon with an outline of several significant developments in colorectal cancer treatment that have affected the care of patients. This review is not intended to report on every important publication of the past few years nor is it intended to be encyclopedic. The author simply hopes to provide a useful reference for surgeons in their daily practice. PMID- 16468545 TI - Evaluation of the potential acuity meter in predicting visual acuity after photodynamic therapy for choroidal neovascularization. AB - BACKGROUND AND OBJECTIVE: To evaluate the potential acuity meter in predicting visual acuity after photodynamic therapy of choroidal neovascular membranes caused by age-related macular degeneration. PATIENTS AND METHODS: Retrospective study of 55 eyes in 51 patients with subfoveal predominantly classic choroidal neovascularization due to age-related macular degeneration. Pre-photodynamic therapy Snellen visual acuity minus the visual acuity with the potential acuity meter was compared with the actual improvement in visual acuity. Significant improvement in visual acuity after photodynamic therapy was defined as a gain of more than 2 lines in Snellen visual acuity (0.2 logarithm of the minimal angle of resolution units). RESULTS: Eighteen (51.4%) of 35 eyes with a potential for improvement experienced an actual improvement in their Snellen visual acuity after photodynamic therapy (P < .001). All eyes lacking a potential for improvement failed to experience an actual improvement in visual acuity after photodynamic therapy. CONCLUSION: The eyes that demonstrated better visual acuity with the potential acuity meter in this study seemed more likely to experience an improvement in visual acuity after photodynamic therapy. PMID- 16468546 TI - Sutureless vitreoretinal surgery using a new 25-gauge transconjunctival system. AB - BACKGROUND AND OBJECTIVE: To evaluate the efficacy of vitreoretinal surgery using a new 25-gauge transconjunctival sutureless vitrectomy system. PATIENTS AND METHODS: Forty-one eyes of 41 consecutive patients were treated from July 2003 to October 2003. Diagnoses included diabetic vitreous hemorrhage (n = 19), diabetic tractional retinal detachment (n = 4), diabetic macular edema (n = 2), macular hole (n = 3), epiretinal membrane (n = 2), branch retinal vein occlusion (n = 3), central retinal vein occlusion (n = 1), vitreous opacity (n = 4), retinal detachment (n = 1), capsular block syndrome (n = 1), and submacular hemorrhage (n = 1). The procedure was combined with phacoemulsification in 14 eyes. RESULTS: All except 3 patients were operated on under retrobulbar anesthesia only. Four cases required the superior sclerotomy to be converted to a 20-gauge procedure. The mean operation time was 33 minutes and the mean balanced salt solution used was 59 cc. The mean visual acuity improved rapidly during 3 to 4 weeks following 25-gauge transconjunctival sutureless vitrectomy from 20/275 to 20/125, and then to 20/100 during the subsequent 5 months. Transient hypotony occurred in only 1 eye. Neither wound leakage nor infection was identified. CONCLUSION: Despite some limitations in surgical indications, 25-gauge transconjunctival sutureless vitrectomy appeared to increase the efficiency of vitrectomy and to facilitate postoperative visual recovery in various vitreoretinal diseases. PMID- 16468547 TI - Posterior capsule opacity and choroidal neovascularization in patients with age related macular degeneration. AB - BACKGROUND AND OBJECTIVE: To present a series of 3 patients with the dry form of age-related macular degeneration (AMD) in whom choroidal neovascularization (CNV) was observed following neodymium: yttrium-aluminum-garnet (Nd:YAG) capsulotomy. PATIENTS AND METHODS: Three consecutive patients aged 80 to 87 years (average = 83.7 years) with hard drusen or retinal pigment epithelial changes underwent uneventful cataract extraction between 6 months and 20 years before undergoing Nd:YAG capsulotomy for posterior capsule opacity. The patients were examined for retinal changes before laser treatment and at regular intervals after treatment. RESULTS: All patients developed CNV between 12 days and 1 month after capsulotomy that caused a decrease in the visual acuity from 20/20-20/40 to 20/200--counting fingers at 4 feet. The fellow eye did not show a substantial change in AMD over a year of follow-up. Two of the fellow eyes had disciform scar due to CNV before the cataract surgery, and these patients remained legally blind. CONCLUSIONS: The development of CNV after Nd:YAG capsulotomy suggests a temporal association between the two and calls for further study. Because posterior capsule opacity may mask CNV, patients with dry AMD and posterior capsule opacity who experience decreased visual acuity or glare may undergo indocyanine green angiography before it is decided to perform Nd:YAG capsulotomy. PMID- 16468548 TI - Clinical manifestations of orbital mass lesions at the Jules Stein Eye Institute, 1999-2003. AB - BACKGROUND AND OBJECTIVE: To describe the clinical manifestations, diagnoses, treatments, and outcomes of orbital mass lesions at a tertiary care referral center. PATIENTS AND METHODS: All cases of newly diagnosed or referred orbital tumors at the Jules Stein Eye Institute from 1999 to 2003 were reviewed retrospectively. Demographic and clinical data were extracted from the electronic oculoplastics registry of the Division of Orbital and Ophthalmic Plastic Surgery. RESULTS: Three hundred sixty-nine cases of orbital mass lesions were evaluated (167 males and 202 females; mean age = 48 years). The most common presenting symptoms were mass/proptosis, pain, swelling, inflammation, and diplopia. The most common categories of diagnosis were cystic or structural lesions, benign tumors, inflammatory processes, neuronal processes, and fibrous processes. Increasing age was associated with an increased incidence of primary and metastatic malignant tumors. Half of all cases required surgical intervention consisting of excision, debulking, or exenteration; 20% to 30% of cases were managed conservatively. CONCLUSIONS: The differential diagnosis of orbital mass lesions differs across age groups. No clinical sign or symptom is specific for the underlying diagnosis and the biological behavior of the abnormal process may be misleading. Therefore, a careful diagnostic approach that considers the benefit of imaging studies must be undertaken. Almost 50% of these mass lesions can be managed with nonsurgical intervention. PMID- 16468549 TI - Results of trabeculectomy with adjunctive intraoperative mitomycin C in Chinese patients with glaucoma. AB - BACKGROUND AND OBJECTIVE: To report the results of trabeculectomy with adjunctive intraoperative mitomycin C in Chinese patients with glaucoma. PATIENTS AND METHODS: The medical records of Chinese patients who received trabeculectomy with intraoperative mitomycin C from 1992 to 1998 were retrospectively reviewed. A "successful" trabeculectomy was defined as one after which the intraocular pressure (IOP) could be controlled to between 5 and 21 mm Hg, inclusive, with no more than 3 glaucoma medications. RESULTS: One hundred fourteen trabeculectomies in 105 eyes of 90 patients were retrospectively reviewed. Mean age +/- standard deviation was 48.1 +/- 21.9 years. Primary open-angle glaucoma (43.0%) and angle closure glaucoma (27.2%) were the most common diagnoses leading to trabeculectomy. The mean follow-up +/- standard deviation was 29.6 +/- 18.6 months (range, 6 to 92 months). The overall success rate at the last follow-up was 73.7% (84 of 114 trabeculectomies). CONCLUSIONS: Trabeculectomy with mitomycin C is a safe and effective procedure for the control of IOP in Chinese patients with glaucoma. PMID- 16468550 TI - Vitrectomy for stage 1 macular holes identified by optical coherence tomography. AB - BACKGROUND AND OBJECTIVE: To describe a series of patients with decreased vision secondary to stage 1 macular hole confirmed by optical coherence tomography (OCT) that failed to spontaneously resolve with observation. To determine whether current surgical techniques can prevent progression to a full-thickness macular hole and lead to improved visual acuity. PATIENTS AND METHODS: Retrospective, comparative case series presenting patient demographics, duration of symptoms, timing of surgery, preoperative and postoperative vision, and clinical outcome based on examination and OCT. RESULTS: Five eyes of five patients with stage 1 macular holes identified on OCT required surgical repair. All eyes showed anatomic closure and lack of progression to a full-thickness hole confirmed by OCT after one procedure. Mean preoperative visual acuity was 20/102 (range, 20/50 to 20/200) and mean postoperative visual acuity was 20/52 (range, 20/25 to 20/200). Average improvement for all five eyes was 3 Snellen lines, with four of the five (80%) improving 5 lines each. CONCLUSION: In symptomatic eyes with stage 1 macular holes that fail to spontaneously resolve with observation, vitrectomy with intraocular gas tamponade may prevent progression to a full-thickness hole and lead to improved visual acuity. PMID- 16468551 TI - Transpupillary thermotherapy alone and enhanced by standard doses of indocyanine green in pigmented rabbits. AB - BACKGROUND AND OBJECTIVE: To evaluate the chorioretinal effects of transpupillary thermotherapy (TTT) and TTT enhanced by standard doses of indocyanine green (ICG) in pigmented rabbits. MATERIALS AND METHODS: In 25 eyes, two retinal areas were irradiated using identical subthreshold diode laser irradiation. The subthreshold diode laser irradiation level was first determined using the other eye. ICG (0.5 mg/kg) was injected before irradiation of the second area. Red-free photographs, fluorescein angiography, and ICG videoangiography were performed after TTT. Specimens were prepared for light microscopy. RESULTS: In 35% of the cases, ICG videoangiography revealed a hypofluorescent spot in the area irradiated after injection of ICG that disappeared after ICG re-injection. None of the areas irradiated before ICG injection were hypofluorescent. Red-free photographs and fluorescein angiography were normal in most cases. Light microscopy demonstrated patent choriocapillaris and choroid in most areas. CONCLUSIONS: Subthreshold TTT, whether enhanced by ICG or not, did not cause significant choroidal or choriocapillaris vascular occlusion. Standard doses of ICG had no significant effect on TTT. PMID- 16468552 TI - Effects of inadequate anterior segment compensation on measurements with scanning laser polarimetry. AB - The effects of poor anterior segment compensation on scanning laser polarimetry measurements of the retinal nerve fiber layer (RNFL) were systematically explored. A prototype scanning laser polarimeter with an adjustable compensator to neutralize anterior segment birefringence was used. By systematically varying the magnitude and axis of anterior segment compensation in a healthy and a glaucomatous eye, marked changes were observed in RNFL appearance: the healthy eye could appear to have glaucomatous damage, whereas the glaucomatous eye could appear to have a thicker and healthier RNFL. Even small amounts of uncompensated corneal birefringence, which may occur in routine clinical use, resulted in apparent changes in RNFL morphology. Knowledge of this effect is important for clinicians when using scanning laser polarimetry in clinical practice. PMID- 16468553 TI - Optical coherence tomography study of optic disc melanocytoma. AB - Optic disc melanocytoma is a well-known clinical entity occurring as a deeply pigmented lesion on the optic nerve head. A 40-year-old man with a pigmented lesion overlying the left optic disc was evaluated with optical coherence tomography. The single high reflectance band from the anterior edge of the disc lesion with optical back shadowing behind on optical coherence tomography helped to pinpoint the diagnosis of the lesion as optic disc melanocytoma. Quantitative assessment of the mass with optical coherence tomography for its growth did not show any increase in size horizontally or vertically after 6 months. Optical coherence tomography is useful for obtaining the exact measurements of the mass lesion and may be used as a tool to follow the progress and growth pattern of the lesion. PMID- 16468554 TI - Intraoperative management of nonfunctioning Ahmed glaucoma valve implant. AB - A 62-year-old woman with neovascular glaucoma underwent filtering surgery with the Ahmed glaucoma valve (New World Medical, Inc., Rancho Cucamonga, CA). Intraoperative hypotony secondary to a defective valve was noted and managed by excising the valve mechanism. The implant was then treated as if it was non valved by placement of an obturator suture. The postoperative course was unremarkable for a non-valved implant. This case demonstrates how intraoperative hypotony secondary to overfiltration of an Ahmed glaucoma valve implant can be successfully managed by converting to a non-valved implant. PMID- 16468555 TI - Management of a displaced angle-supported anterior chamber intraocular lens. AB - A 49-year-old patient had a displacement of an anterior chamber intraocular lens (IOL) producing a cyclodialysis cleft and persistent hypotony with maculopathy. The patient underwent intraocular lens extraction, repair of the cyclodialysis cleft, and iris-fixation of an acrylic posterior chamber IOL with resolution of the hypotony and improvement of the maculopathy and best-corrected visual acuity. Many surgeons consider the implantation of an anterior chamber IOL as their first choice in the treatment of inadequate capsule support associated with cataract surgery, probably because it is a relatively easy procedure to perform. However, insufficient preoperative knowledge of the anterior segment biometry can lead to inadequate lens size and postoperative complications. PMID- 16468556 TI - Management of bilateral ectopia lentis et pupillae syndrome. AB - A 52-year-old patient presented with signs clinically consistent with ectopia lentis et pupillae syndrome. The patient was treated successfully with vitrectomy, dislocated lens removal using perfluorocarbon liquid and phacofragmentation in the vitreous cavity, pupil reconstruction, and scleral fixated intraocular lens implantation in both eyes. Despite the fact that the surgery was successful in technical terms, the final visual outcome was not as good as expected. This was caused by the optic nerve atrophy resulting from long lasting glaucoma. Nevertheless, the described surgical techniques may be considered an effective method of treatment in cases of ectopia lentis et pupillae syndrome. PMID- 16468557 TI - Resolution of macular scotoma after radial optic neurotomy in central retinal vein occlusion. AB - Radial optic neurotomy was performed on a hypertensive 65-year-old man with decreased vision in the right eye secondary to ischemic central retinal vein occlusion. Three months later, his visual acuity improved to 20/50. There was improvement in disc and macular edema, and reduction in retinal hemorrhages clinically and angiographically. Liquid crystal display microperimetry revealed improvement in fixation stability and macular sensitivity. Radial optic neurotomy appears to be an effective treatment alternative for selected patients with ischemic central retinal vein occlusion, and results in rapid improvement in visual acuity, central fixation, and macular scotoma as measured by a liquid crystal display microperimeter. Liquid crystal display microperimetry is a good and reliable alternative to the scanning laser ophthalmoscope for macular perimetry. PMID- 16468558 TI - Persistent choroidal thickening despite photodynamic therapy for circumscribed choroidal hemangioma. AB - A 19-year-old man presented with blurred vision in the right eye, which was diagnosed as a circumscribed choroidal hemangioma. The choroidal thickening on B scan ultrasonography persisted after receiving repeat photodynamic therapy with verteporfin, although there was regression of the tumor. PMID- 16468560 TI - Minimally invasive method for repair of rhegmatogenous retinal detachment following treatment for retinoblastoma. AB - A minimally invasive method for repair of rhegmatogenous retinal detachment following treatment of retinoblastoma is described. Two children with retinoblastoma developed rhegmatogenous retinal detachments following treatment with a combination of chemotherapy and cryotherapy. Each eye underwent a non drainage scleral buckling procedure without retinopexy of the retinal break. Postoperatively, the retinas reattached and retinopexy with indirect laser photocoagulation was performed during subsequent examinations under anesthesia. The retinas have remained attached with 2 to 5 years of follow-up. The tumor recurred in case 2, was treated with additional chemotherapy, and has undergone type 1 regression. Non-drainage scleral buckling without retinopexy is a useful technique for repairing rhegmatogenous retinal detachment in eyes with retinoblastoma. The retinal break can be treated postoperatively with indirect laser photocoagulation to minimize the chance of viable tumor cell dissemination. PMID- 16468559 TI - Laser ablation of an enlarging small melanocytic choroidal tumor: 16-year follow up and rationale for treatment. AB - Results of laser photocoagulation of a growing pigmented choroidal lesion for which the differential diagnosis included enlarging nevus and small melanoma and the rationale for such treatment are described. A 37-year-old woman underwent ablative laser photocoagulation for a small juxtapapillary pigmented choroidal lesion that was documented photographically to enlarge in diameter from 300 to 1,750 microns after 8 years. Sixteen years after laser photocoagulation, the visual acuity remains 20/20 with no further growth or recurrence of the lesion. Local treatment may be justified for a small pigmented choroidal lesion that is documented to grow. PMID- 16468561 TI - Lenticonus revisited. PMID- 16468562 TI - TSK fuzzy systems types II and III stability analysis: continuous case. AB - We propose a new approach for the stability analysis of continuous Sugeno Types II and III dynamic fuzzy systems. We introduce the concept of fuzzy positive definite and fuzzy negative definite systems and use them in arguments similar to those of traditional Lyapunov stability theory to derive new conditions for stability and asymptotic stability for continuous Type II/III dynamic fuzzy systems. To demonstrate the new approach, we apply it to numerical examples. PMID- 16468563 TI - Switching fuzzy controller design based on switching Lyapunov function for a class of nonlinear systems. AB - This paper presents a switching fuzzy controller design for a class of nonlinear systems. A switching fuzzy model is employed to represent the dynamics of a nonlinear system. In our previous papers, we proposed the switching fuzzy model and a switching Lyapunov function and derived stability conditions for open-loop systems. In this paper, we design a switching fuzzy controller. We firstly show that switching fuzzy controller design conditions based on the switching Lyapunov function are given in terms of bilinear matrix inequalities, which is difficult to design the controller numerically. Then, we propose a new controller design approach utilizing an augmented system. By introducing the augmented system which consists of the switching fuzzy model and a stable linear system, the controller design conditions based on the switching Lyapunov function are given in terms of linear matrix inequalities (LMIs). Therefore, we can effectively design the switching fuzzy controller via LMI-based approach. A design example illustrates the utility of this approach. Moreover, we show that the approach proposed in this paper is available in the research area of piecewise linear control. PMID- 16468564 TI - Genetic algorithm based methodology for breaking the steganalytic systems. AB - Steganalytic techniques are used to detect whether an image contains a hidden message. By analyzing various image features between stego-images (the images containing hidden messages) and cover-images (the images containing no hidden messages), a steganalytic system is able to detect stego-images. In this paper, we present a new concept of developing a robust steganographic system by artificially counterfeiting statistic features instead of the traditional strategy by avoiding the change of statistic features. We apply genetic algorithm based methodology by adjusting gray values of a cover-image while creating the desired statistic features to generate the stego-images that can break the inspection of steganalytic systems. Experimental results show that our algorithm can not only pass the detection of current steganalytic systems, but also increase the capacity of the embedded message and enhance the peak signal-to noise ratio of stego-images. PMID- 16468565 TI - Highly scalable and robust rule learner: performance evaluation and comparison. AB - Business intelligence and bioinformatics applications increasingly require the mining of datasets consisting of millions of data points, or crafting real-time enterprise-level decision support systems for large corporations and drug companies. In all cases, there needs to be an underlying data mining system, and this mining system must be highly scalable. To this end, we describe a new rule learner called DataSqueezer. The learner belongs to the family of inductive supervised rule extraction algorithms. DataSqueezer is a simple, greedy, rule builder that generates a set of production rules from labeled input data. In spite of its relative simplicity, DataSqueezer is a very effective learner. The rules generated by the algorithm are compact, comprehensible, and have accuracy comparable to rules generated by other state-of-the-art rule extraction algorithms. The main advantages of DataSqueezer are very high efficiency, and missing data resistance. DataSqueezer exhibits log-linear asymptotic complexity with the number of training examples, and it is faster than other state-of-the art rule learners. The learner is also robust to large quantities of missing data, as verified by extensive experimental comparison with the other learners. DataSqueezer is thus well suited to modern data mining and business intelligence tasks, which commonly involve huge datasets with a large fraction of missing data. PMID- 16468566 TI - A multiagent evolutionary algorithm for constraint satisfaction problems. AB - With the intrinsic properties of constraint satisfaction problems (CSPs) in mind, we divide CSPs into two types, namely, permutation CSPs and nonpermutation CSPs. According to their characteristics, several behaviors are designed for agents by making use of the ability of agents to sense and act on the environment. These behaviors are controlled by means of evolution, so that the multiagent evolutionary algorithm for constraint satisfaction problems (MAEA-CSPs) results. To overcome the disadvantages of the general encoding methods, the minimum conflict encoding is also proposed. Theoretical analyzes show that MAEA-CSPs has a linear space complexity and converges to the global optimum. The first part of the experiments uses 250 benchmark binary CSPs and 79 graph coloring problems from the DIMACS challenge to test the performance of MAEA-CSPs for nonpermutation CSPs. MAEA-CSPs is compared with six well-defined algorithms and the effect of the parameters is analyzed systematically. The second part of the experiments uses a classical CSP, n-queen problems, and a more practical case, job-shop scheduling problems (JSPs), to test the performance of MAEA-CSPs for permutation CSPs. The scalability of MAEA-CSPs along n for n-queen problems is studied with great care. The results show that MAEA-CSPs achieves good performance when n increases from 10(4) to 10(7), and has a linear time complexity. Even for 10(7) queen problems, MAEA-CSPs finds the solutions by only 150 seconds. For JSPs, 59 benchmark problems are used, and good performance is also obtained. PMID- 16468567 TI - Interactive segmentation of the cerebral lobes with fuzzy inference in 3T MR images. AB - Measurement of volume and surface area of the frontal, parietal, temporal and occipital lobes from magnetic resonance (MR) images shows promise as a method for use in diagnosis of dementia. This article presents a novel computer-aided system for automatically segmenting the cerebral lobes from 3T human brain MR images. Until now, the anatomical definition of cerebral lobes on the cerebral cortex is somewhat vague for use in automatic delineation of boundary lines, and there is no definition of cerebral lobes in the interior of the cerebrum. Therefore, we have developed a new method for defining cerebral lobes on the cerebral cortex and in the interior of the cerebrum. The proposed method determines the boundaries between the lobes by deforming initial surfaces. The initial surfaces are automatically determined based on user-given landmarks. They are smoothed and deformed so that the deforming boundaries run along the hourglass portion of the three-dimensional shape of the cerebrum with fuzzy rule-based active contour and surface models. The cerebrum is divided into the cerebral lobes according to the boundaries determined using this method. The reproducibility of our system with a given subject was assessed by examining the variability of volume and surface area in three healthy subjects, with measurements performed by three beginners and one expert user. The experimental results show that our system segments the cerebral lobes with high reproducibility. PMID- 16468568 TI - Output convergence analysis for a class of delayed recurrent neural networks with time-varying inputs. AB - This paper studies the output convergence of a class of recurrent neural networks with time-varying inputs. The model of the studied neural networks has different dynamic structure from that in the well known Hopfield model, it does not contain linear terms. Since different structures of differential equations usually result in quite different dynamic behaviors, the convergence of this model is quite different from that of Hopfield model. This class of neural networks has been found many successful applications in solving some optimization problems. Some sufficient conditions to guarantee output convergence of the networks are derived. PMID- 16468569 TI - A real-time automated system for the recognition of human facial expressions. AB - A fully automated, multistage system for real-time recognition of facial expression is presented. The system uses facial motion to characterize monochrome frontal views of facial expressions and is able to operate effectively in cluttered and dynamic scenes, recognizing the six emotions universally associated with unique facial expressions, namely happiness, sadness, disgust, surprise, fear, and anger. Faces are located using a spatial ratio template tracker algorithm. Optical flow of the face is subsequently determined using a real-time implementation of a robust gradient model. The expression recognition system then averages facial velocity information over identified regions of the face and cancels out rigid head motion by taking ratios of this averaged motion. The motion signatures produced are then classified using Support Vector Machines as either nonexpressive or as one of the six basic emotions. The completed system is demonstrated in two simple affective computing applications that respond in real time to the facial expressions of the user, thereby providing the potential for improvements in the interaction between a computer user and technology. PMID- 16468570 TI - Genetic programming for simultaneous feature selection and classifier design. AB - This paper presents an online feature selection algorithm using genetic programming (GP). The proposed GP methodology simultaneously selects a good subset of features and constructs a classifier using the selected features. For a c-class problem, it provides a classifier having c trees. In this context, we introduce two new crossover operations to suit the feature selection process. As a byproduct, our algorithm produces a feature ranking scheme. We tested our method on several data sets having dimensions varying from 4 to 7129. We compared the performance of our method with results available in the literature and found that the proposed method produces consistently good results. To demonstrate the robustness of the scheme, we studied its effectiveness on data sets with known (synthetically added) redundant/bad features. PMID- 16468571 TI - Some properties of the weighted OWA operator. AB - Based on the researches on ordered weighted average (OWA) operator, the weighted OWA operator (WOWA) and especially the quantifier guided aggregation method, with the generating function representation of regular increasing monotone (RIM) quantifier technique, we discuss the properties of WOWA operator with RIM quantifier in the respect of orness. With the continuous OWA and WOWA ideas recently proposed by Yager, an improvement on the continuous OWA and WOWA operator is proposed. The properties of WOWA are also extended from discrete to the continuous case. Based on these properties, two families of parameterized RIM quantifiers for WOWA operator are proposed, which have exponential generating function and piecewise linear generating function respectively. One interesting property of these two kinds of RIM quantifiers is that for any aggregated set (or variable) under any weighted (distribution) function, the aggregation values are always consistent with the orness (optimistic) levels, so they can be used to represent the decision maker's preference, and we can get the preference value of fuzzy sets or random variables with the orness level of RIM quantifier as their control parameter. PMID- 16468572 TI - Learning tactical human behavior through observation of human performance. AB - It is widely accepted that the difficulty and expense involved in acquiring the knowledge behind tactical behaviors has been one limiting factor in the development of simulated agents representing adversaries and teammates in military and game simulations. Several researchers have addressed this problem with varying degrees of success. The problem mostly lies in the fact that tactical knowledge is difficult to elicit and represent through interactive sessions between the model developer and the subject matter expert. This paper describes a novel approach that employs genetic programming in conjunction with context-based reasoning to evolve tactical agents based upon automatic observation of a human performing a mission on a simulator. In this paper, we describe the process used to carry out the learning. A prototype was built to demonstrate feasibility and it is described herein. The prototype was rigorously and extensively tested. The evolved agents exhibited good fidelity to the observed human performance, as well as the capacity to generalize from it. PMID- 16468573 TI - Classification of adaptive memetic algorithms: a comparative study. AB - Adaptation of parameters and operators represents one of the recent most important and promising areas of research in evolutionary computations; it is a form of designing self-configuring algorithms that acclimatize to suit the problem in hand. Here, our interests are on a recent breed of hybrid evolutionary algorithms typically known as adaptive memetic algorithms (MAs). One unique feature of adaptive MAs is the choice of local search methods or memes and recent studies have shown that this choice significantly affects the performances of problem searches. In this paper, we present a classification of memes adaptation in adaptive MAs on the basis of the mechanism used and the level of historical knowledge on the memes employed. Then the asymptotic convergence properties of the adaptive MAs considered are analyzed according to the classification. Subsequently, empirical studies on representatives of adaptive MAs for different type-level meme adaptations using continuous benchmark problems indicate that global-level adaptive MAs exhibit better search performances. Finally we conclude with some promising research directions in the area. PMID- 16468574 TI - A functional-based segmentation of human body scans in arbitrary postures. AB - This paper presents a general framework that aims to address the task of segmenting three-dimensional (3-D) scan data representing the human form into subsets which correspond to functional human body parts. Such a task is challenging due to the articulated and deformable nature of the human body. A salient feature of this framework is that it is able to cope with various body postures and is in addition robust to noise, holes, irregular sampling and rigid transformations. Although whole human body scanners are now capable of routinely capturing the shape of the whole body in machine readable format, they have not yet realized their potential to provide automatic extraction of key body measurements. Automated production of anthropometric databases is a prerequisite to satisfying the needs of certain industrial sectors (e.g., the clothing industry). This implies that in order to extract specific measurements of interest, whole body 3-D scan data must be segmented by machine into subsets corresponding to functional human body parts. However, previously reported attempts at automating the segmentation process suffer from various limitations, such as being restricted to a standard specific posture and being vulnerable to scan data artifacts. Our human body segmentation algorithm advances the state of the art to overcome the above limitations and we present experimental results obtained using both real and synthetic data that confirm the validity, effectiveness, and robustness of our approach. PMID- 16468575 TI - FITSK: online local learning with generic fuzzy input Takagi-Sugeno-Kang fuzzy framework for nonlinear system estimation. AB - Existing Takagi-Sugeno-Kang (TSK) fuzzy models proposed in the literature attempt to optimize the global learning accuracy as well as to maintain the interpretability of the local models. Most of the proposed methods suffer from the use of offline learning algorithms to globally optimize this multi-criteria problem. Despite the ability to reach an optimal solution in terms of accuracy and interpretability, these offline methods are not suitably applicable to learning in adaptive or incremental systems. Furthermore, most of the learning methods in TSK-model are susceptible to the limitation of the curse-of dimensionality. This paper attempts to study the criteria in the design of TSK models. They are: 1) the interpretability of the local model; 2) the global accuracy; and 3) the system dimensionality issues. A generic framework is proposed to handle the different scenarios in this design problem. The framework is termed the generic fuzzy input Takagi-Sugeno-Kang fuzzy framework (FITSK). The FITSK framework is extensible to both the zero-order and the first-order FITSK models. A zero-order FITSK model is suitable for the learning of adaptive system, and the bias-variance of the system can be easily controlled through the degree of localization. On the other hand, a first-order FITSK model is able to achieve higher learning accuracy for nonlinear system estimation. A localized version of recursive least-squares algorithm is proposed for the parameter tuning of the first-order FITSK model. The local recursive least-squares is able to achieve a balance between interpretability and learning accuracy of a system, and possesses greater immunity to the curse-of-dimensionality. The learning algorithms for the FITSK models are online, and are readily applicable to adaptive system with fast convergence speed. Finally, a proposed guideline is discussed to handle the model selection of different FITSK models to tackle the multi-criteria design problem of applying the TSK-model. Extensive simulations were conducted using the proposed FITSK models and their learning algorithms; their performances are encouraging when benchmarked against other popular fuzzy systems. PMID- 16468576 TI - A study of evolutionary multiagent models based on symbiosis. AB - Multiagent Systems with Symbiotic Learning and Evolution (Masbiole) has been proposed and studied, which is a new methodology of Multiagent Systems (MAS) based on symbiosis in the ecosystem. Masbiole employs a method of symbiotic learning and evolution where agents can learn or evolve according to their symbiotic relations toward others, i.e., considering the benefits/losses of both itself and an opponent. As a result, Masbiole can escape from Nash Equilibria and obtain better performances than conventional MAS where agents consider only their own benefits. This paper focuses on the evolutionary model of Masbiole, and its characteristics are examined especially with an emphasis on the behaviors of agents obtained by symbiotic evolution. In the simulations, two ideas suitable for the effective analysis of such behaviors are introduced; "Match Type Tile world (MTT)" and "Genetic Network Programming (GNP)". MTT is a virtual model where tile-world is improved so that agents can behave considering their symbiotic relations. GNP is a newly developed evolutionary computation which has the directed graph type gene structure and enables to analyze the decision making mechanism of agents easily. Simulation results show that Masbiole can obtain various kinds of behaviors and better performances than conventional MAS in MTT by evolution. PMID- 16468577 TI - On image matrix based feature extraction algorithms. AB - Principal component analysis (PCA) and linear discriminant analysis (LDA) are two important feature extraction methods and have been widely applied in a variety of areas. A limitation of PCA and LDA is that when dealing with image data, the image matrices must be first transformed into vectors, which are usually of very high dimensionality. This causes expensive computational cost and sometimes the singularity problem. Recently two methods called two-dimensional PCA (2DPCA) and two-dimensional LDA (2DLDA) were proposed to overcome this disadvantage by working directly on 2-D image matrices without a vectorization procedure. The 2DPCA and 2DLDA significantly reduce the computational effort and the possibility of singularity in feature extraction. In this paper, we show that these matrices based 2-D algorithms are equivalent to special cases of image block based feature extraction, i.e., partition each image into several blocks and perform standard PCA or LDA on the aggregate of all image blocks. These results thus provide a better understanding of the 2-D feature extraction approaches. PMID- 16468578 TI - A note on the spread of worms in scale-free networks. AB - This paper considers the spread of worms in computer networks using insights from epidemiology and percolation theory. We provide three new results. The first result refines previous work showing that epidemics occur in scale-free graphs more easily because of their structure. We argue, using recent results from random graph theory that for scaling factors between 0 and approximately 3.4875, any computer worm infection of a scale-free network will become an epidemic. Our second result uses this insight to provide a mathematical explanation for the empirical results of Chen and Carley, who demonstrate that the Countermeasure Competing strategy can be more effective for immunizing networks to viruses or worms than traditional approaches. Our third result uses random graph theory to contradict the current supposition that, for very large networks, monocultures are necessarily more susceptible than diverse networks to worm infections. PMID- 16468579 TI - Robust and fast learning for fuzzy cerebellar model articulation controllers. AB - In this paper, the online learning capability and the robust property for the learning algorithms of cerebellar model articulation controllers (CMAC) are discussed. Both the traditional CMAC and fuzzy CMAC are considered. In the study, we find a way of embeding the idea of M-estimators into the CMAC learning algorithms to provide the robust property against outliers existing in training data. An annealing schedule is also adopted for the learning constant to fulfill robust learning. In the study, we also extend our previous work of adopting the credit assignment idea into CMAC learning to provide fast learning for fuzzy CMAC. From demonstrated examples, it is clearly evident that the proposed algorithm indeed has faster and more robust learning. In our study, we then employ the proposed CMAC for an online learning control scheme used in the literature. In the implementation, we also propose to use a tuning parameter instead of a fixed constant to achieve both online learning and fine-tuning effects. The simulation results indeed show the effectiveness of the proposed approaches. PMID- 16468580 TI - Exponential synchronization of a class of neural networks with time-varying delays. AB - This paper aims to present a synchronization scheme for a class of delayed neural networks, which covers the Hopfield neural networks and cellular neural networks with time-varying delays. A feedback control gain matrix is derived to achieve the exponential synchronization of the drive-response structure of neural networks by using the Lyapunov stability theory, and its exponential synchronization condition can be verified if a certain Hamiltonian matrix with no eigenvalues on the imaginary axis. This condition can avoid solving an algebraic Riccati equation. Both the cellular neural networks and Hopfield neural networks with time-varying delays are given as examples for illustration. PMID- 16468581 TI - Robust H infinity static output feedback control of fuzzy systems: an ILMI approach. AB - This paper examines the problem of robust H infinity static output feedback control of a Takagi-Sugeno fuzzy system. The proposed robust H infinity static output feedback controller guarantees the pounds 2 gain of the mapping from the exogenous disturbances to the regulated output to be less than or equal to a prescribed level. The existence of a robust H infinity static output feedback control is given in terms of the solvability of bilinear matrix inequalities. An iterative algorithm based on the linear matrix inequality is developed to compute robust H infinity static output feedback gains. To reduce the conservatism of the design, the structural information of membership function characteristics is incorporated. A numerical example is used to illustrate the validity of the design methodologies. PMID- 16468582 TI - A sound and complete fuzzy temporal constraint logic. AB - In this work, we define an extended fuzzy temporal constraint logic (EFTCL) based on possibilistic logic. EFTCL allows us to handle fuzzy temporal constraints between temporal variables and, therefore, enables us to express interrelated events through fuzzy temporal constraints. EFTCL is compatible with a theoretical temporal reasoning model: the fuzzy temporal constraint networks (FTCN). The syntax, the semantics and the deduction and refutation theorems for EFTCL are similar to those defined for the sound and noncomplete fuzzy temporal constraint logic (FTCL). In this paper, a resolution principle for performing inferences which take these constraints into account is proposed for EFTCL. Moreover, we prove the soundness and the completeness of the refutation by resolution in EFTCL. PMID- 16468583 TI - Analysis of a master-slave architecture for distributed evolutionary computations. AB - This paper introduces a new mathematical model of the master-slave architecture for distributed evolutionary computations (EC). This model is validated using a concrete implementation based on the Distributed BEAGLE C++ framework. Results show that contrary to (current) popular belief, master-slave architectures are able to scale well over local area networks of workstations using off-the-shelf networking equipment. The main properties of the master-slave are also compared with those of the more mainstream island-model. PMID- 16468584 TI - The road less traveled: providing home care in rural communities. PMID- 16468585 TI - Rural home health care initiative. Transportation. PMID- 16468586 TI - All roads lead to quality at OSF Home Care. AB - Technology and accessibility play important roles in healthcare today, improving service to all patients, both rural and non-rural. According to OSF Home Care Administrator, Katie Jones, "The goal of home care is to serve patients in the comfort of their homes, no matter what roads we need to travel to get there." PMID- 16468587 TI - Telemanagement in hospice-a new frontier. PMID- 16468588 TI - Home health and hospice in rural America. PMID- 16468589 TI - Work together, win together: negotiating better contract reimbursement. PMID- 16468590 TI - Are we really prepared? Since 9/11, where has the urgency gone? AB - From this study, it was determined that all agencies have updated their Emergency Preparedness Plans. The recommendation to provide Universal ID Cards for home health care workers has not as yet been implemented. And, in conclusion of this part of the study, most agencies have not increased inter-agency cooperation. The section that discussed the recommendation to improve communication and transportation systems, due to the comprehensiveness of this subject matter, will be found in the next edition of CARING--Are We Really Prepared? Part II. In addition, an overall summary of all emergency preparedness recommendations will be discussed at that time. PMID- 16468591 TI - The mosaic of Canadian home care. PMID- 16468593 TI - The "state" of home care & hospice. AB - There is at least one home care and hospice industry association in each of our 50 United States and Puerto Rico. These associations represent tens of thousands of home care and hospice providers. They provide state and local leadership, professional and para-professional training; and other services designed to help home care and hospice providers care for our nation's sick, disabled and elderly. These are their stories. PMID- 16468592 TI - Home care in Lebanon. PMID- 16468594 TI - New hospice regulations allow discharge for cause: require clinical documentation for certification. Final rule effective January 23, 2006. PMID- 16468595 TI - The benefits and challenges of technology in the delivery of home health care in rural America. PMID- 16468596 TI - Selling to the "sticks". PMID- 16468598 TI - Pay for performance: great idea. But can it deliver great results? PMID- 16468597 TI - Q & A. You ask and Sam answers. PMID- 16468599 TI - Accelerated unlearning. PMID- 16468600 TI - The future of geriatrics. PMID- 16468601 TI - Telemonitoring technology: changing lives and changing home care. PMID- 16468602 TI - Caring thoughts. Bringing health care to America's frontier. PMID- 16468603 TI - [The legal and occupational health aspects of B and C type viral hepatitis]. AB - INTRODUCTION: Viral hepatitis is a problem of population health. Recent data show an estimated number of 400 million patients infected with hepatitis B and 170 million with hepatitis C virus in the world. The prevalence of HBV infected patients is estimated to 50000, the prevalence of the HCV patients to 70,000 in Hungary. AIMS: The authors report the legal and occupational health aspects of viral hepatitis. The knowledge of these facts could help both the patients and doctors. This paper is on the employment, occupational restrictions of the infected patients, the legal aspects of the discrimination and the vaccination. The authors report the knowledge, rules and restrictions relating to health service employees in a separate subchapter. They call the attention to the occupational health aspects in relation to the infected patients and health service employees from the point of view of the occupational health specialist, the commissioner responsible for safeguarding the patients' interest and the specialist attending the infected patients. PMID- 16468604 TI - [Continuing care of patients with cardiovascular risk in general practice: patients with dyslipidemia and their care]. AB - INTRODUCTION: The role of dyslipidemia is essential in the development of atherosclerosis, therefore continuing care of dyslipidemic patients is an extremely important task in cardiovascular prevention. AIMS: This study was aimed to investigate continuing care of dyslipidemic patients in general practices. Method of the study was a questionnaire survey, 397 patients of 39 general practices were involved into the study. RESULTS: The cause of their continuing care was hypercholesterolemia in 91.7%, hypertriglyceridemia in 69%, decreased HDL-C in 18.4%, and these alterations frequently occurred together. Hypertension in 77%, disturbances of carbohydrate metabolism in 41.7%, increased BMI in 43.2% were found. Patients were smokers in 26.5%, given up smoking in 13.2%. They had cardiovascular problems in 56.5%, cerebrovascular problems in 18.6% and peripheral vascular ones in 20.3%. Patients were treated with lipid lowering drugs in 87.7%, statins were used in 81.7%, fibrates in 29.6% and both of them in 11.5%. The main results of continuing care were: in the whole group frequency of cholesterol level > 5.2 mmol/l decreased by 13%, frequency of triglyceride level >1.7 mmol/l decreased by 4%; cholesterol target values were reached in 3% of patients with high cardiovascular risk, in 22.2% of patients with medium risk, and in 66.7% of patients with mild risk. CONCLUSIONS: These results suggest that there is a need for more intensive drug treatment of dyslipidemic patients to reach the target lipid levels, and for regular assessment of continuing care of these patients. PMID- 16468605 TI - [Cultural history of wine, the theoretical background of wine therapy]. AB - The knowledge of wine and grapes as old as the cultural history of human population. The small consumption of wine could be advantageous. Also it has been well established for several thousands of years, that it could cause acute and chronic injuries after higher consumption. Its antiinfectious effect in dermatological drugs had been known in the archeological ages. The unguents containing wine polyphenols are advantageous in some dermatological injuries and also in the treatment of muscular and articular alterations. Moderate consumption of wine may decrease the mortality rate of cardiovascular diseases. In healthy individuals consumption of one-two dl of wine can decrease the mortality rate of cardiovascular diseases. This quantity, however, may be injurious in pregnancy, in children, and in various organic diseases, especially in liver disease, as well as if combined with common drugs. PMID- 16468606 TI - [Abdominal partial resection of the vagina and colpopexy: experiences with the procedure for posthysterectomy vault prolapse in 74 cases]. AB - OBJECTIVE: To assess the abdominal partial resection of the vagina and infundibulopelvic colpopexia in women with posthysterectomy vaginal vault prolapse. OPERATIVE PROCEDURE AND MATERIAL: Slightly modifying the method described by Lampe, after laparotomy the elongated vaginal wall is resected and the vaginal vault is fixed by the sutures placed into the infundibulopelvic, sacrouterine, and round ligaments. The area is covered and elevated by the overlapping peritoneum. During the last 15 years (July 1990 - July 2005) the procedure was offered and performed in 74 women because of vaginal eversion (aged 28 to 84 years; average age at operation was 58.5 years) after abdominal (24 cases) or vaginal (39 cases) hysterectomy or supravaginal amputation (4 cases) or abdominal colpopexy (7 cases). In 16 cases, anterior or posterior colporrhaphy were subsequently performed because of cystocele or rectocele or both. RESULTS: Perioperative complications included two bladder injury (2.6%), and transitory voiding difficulty in five cases (6.7%). There was neither bowel nor ureter injury. Patients were followed up annually by pelvic examination; in one of the 74 patients the vaginal eversion partly relapsed and the colpopexy was repeated. In one patient 18 months later because of intraabdominal adhesions a laparotomy and adhesiolysis was performed. All patients have a functional vagina without urinary incontinence and without pelvic pain or any pelvic discomfort. CONCLUSION: The abdominal partial resection of the vagina and colpopexy to the pelvic ligaments seems simpler than other techniques that are commonly used and a safe and reliable operation for the correction of posthysterectomy vault prolapse and enterocele. A long term follow up is necessary to detect any late complications. The operation should be made only by gynecologists trained in the surgery of pelvic retroperitoneum. PMID- 16468607 TI - [Collagen type IV nephropathy: from thin basement membrane nephropathy to Alport syndrome]. AB - INTRODUCTION: Collagen type IV nephropathy includes the Goodpasture syndrome, thin basement membrane nephropathy and the Alport syndrome. Mutations in the coding Col(IV)A3/A4 and Col(IV)A5 genes are probable causes of the latter two. Thin basement membrane nephropathy is mostly familial and has an autosomal dominant inheritance, at least 40% of the families have hematuria that co segregates with the Col(IV)A3 and/or Col(IV)A4 loci. 85% of Alport syndrome cases are transmitted as an X-linked semidominant form due to Col(IV)A5 mutations. About 14% of Alport syndrome cases exhibit autosomal recessive, and 1% autosomal dominant inheritance, both caused by mutations in the Col(IV)A3 or Col(IV)A4 genes in boys and in girls. AIM: The co-segregation pattern of hematuria was examined in two families with thin basement membrane nephropathy and one family with the Alport syndrome, using short tandem repeat markers, spanning the Col(IV)A3/A4 and Col(IV)A5 loci to assess their linkage to the clinical symptoms and morphological alterations in the renal biopsy specimens. METHODS: Markers: Col(IV)A3: CAll and D2S401; Col(IV)A4: HaeIII/RFLP; and Col(IV)A5: DXS456, 2B6 and 2B20. RESULTS: The hematuria displayed autosomal dominant inheritance and co segregated with Col(IV)A3 markers in one of the thin basement membrane nephropathy families. In the second, the hematuria did not segregate with the Col(IV)A3/A4 or Col(IV)A5 loci, suggesting the possibility of another genetic locus for the disease. The Alport syndrome exhibited autosomal recessive inheritance and did not link to Col(IV)A5 markers, and the Col(IV)A3/A4 markers were informative only in part. CONCLUSION: Knowledge of the inheritance and genetic background of collagen type IV nephropathy will be very important in the diagnostics and genetic counseling in the future. PMID- 16468610 TI - [In memoriam Viktor Deutsch (1926-1984)]. PMID- 16468609 TI - [In memoriam Janos Loessl (1887-1955)]. PMID- 16468608 TI - [A pioneer of medical numismatics]. PMID- 16468611 TI - [The Human Genome Project, genetic viability and genetic epidemiology]. AB - The goal of the Human Genome Project to elucidate the complete sequence of the human genome has been achieved. The aims of the "post-genome" era are explaining the genetic information, characterisation of functional elements encoded in the human genome and mapping the human genetic variability as well. Two unrelated human beings also share 99.9% of their genomic sequence. The difference of 0.1% is the result of genetic polymorphisms: single nucleotide polymorphisms, repetitive sequences and insertion/deletion. The genetic differences, coupled with environmental exposures will determine the phenotypic variation we observe in health or disease. The disease-causing genetic variants can be identified by linkage analysis or association studies. The knowledge of human genome and application of multiple biomarkers will improve our ability to identify individuals at risk, so that preventive interventions can be applied, earlier diagnosis can be made and treatment can be optimized. PMID- 16468612 TI - [Continuing care of patients with cardiovascular risk in general practice: diabetic patients and their care]. AB - INTRODUCTION: Both types of diabetes mellitus are conditions with high cardiovascular risk. AIMS: This work was aimed to study the frequency of cardiovascular risk factors, macrovascular and microvascular complications and to assess the results of continuous diabetes care in an adult population with type 1 and type 2 diabetes. Method of the study was a questionnaire survey, altogether 400 patients, 49 with type 1 and 351 with type 2 diabetes, were enrolled to it. RESULTS: Frequency of cardiovascular complications, obesity, hypertension and dyslipidemia was higher in type 2 diabetes, smoking and microvascular complications were more frequent in type 1 diabetes. The ratio of conservative intensive and conventional insulin therapy in type 1 diabetes was 70% and 30%, respectively. Treatment methods used in type 2 diabetes were: diet only: 8%, oral antidiabetic therapy: 78%, antidiabetic agent and insulin: 7%, insulin therapy 7%. Ratios of the patients having at least three laboratory results were: fasting blood glucose: 50%, postprandial blood glucose: 30%, haemoglobin A(1C): 10%. Ratios of patients reached the target results were in type 1 and type 2 diabetes were: fasting blood glucose: 27% vs. 14%, postprandial blood glucose: 26% vs. 18%, haemoglobin-A: 29% vs. 34%. Ratios of the patients in micro- and macrovascular risk category were: fasting blood glucose: 59% vs. 68%, postprandial blood glucose: 54% and 53%, haemoglobin-A(1C): 40% vs. 27%. CONCLUSIONS: Frequency of investigations to estimate glycemic control as well as ratio of patients reached target values were rather low, ratio of patients in micro- and macrovascular risk category was high. For these reasons there is a need for a more intensive continuous diabetes care to reach better results. PMID- 16468613 TI - [Noninvasive assessment of endothelial function in hemodialyzed hypertensive patients by laser Doppler flowmetry]. AB - BACKGROUND: End stage renal disease and hypertension are associated with higher cardiovascular mortality. Endothelial dysfunction plays an important role in the pathogenesis of cardiovascular diseases. The authors investigated the endothelium dependent and -independent vasodilation in the forearm skin microcirculation and the plasma markers of endothelial damage in hypertensive hemodialysed patients and in normotensive control subjects. METHODS: Laser Doppler flowmetry with iontophoresis of acetylcholine and sodium nitroprusside and the postocclusive reactive hyperemia test was performed in 22 normal control subjects and in 21 hemodialysed patients with hypertension. Levels of endothelin-1, big-endothelin, and von Willebrand Factor were measured, as well. RESULTS: The average hyperemic response to the two doses of acetylcholine iontophoresis was 474 +/- 83%; 836 +/- 97% in the control subjects, and 160 +/- 26%; 360 +/- 67% in the hemodialysed patients group (p < 0.05). The vasodilation after the two doses of sodium nitroprusside was 381 +/- 60%, 782 +/- 81% in the control group and 186 +/- 42%; 379 +/- 63% in the dialysed patients group (p < 0.05 compared to control, respectively). The average peak flow during the postocclusive reactive hyperemia test was significantly lower in hemodialysed hypertensives (234 +/- 48%) compared to healthy control subjects (434 +/- 36%, p < 0.05). Levels of endothelin-1, big endothelin, von Willebrand Factor and von Willebrand Factor activity were significantly higher in the patient group compared to the control subjects. CONCLUSIONS: In hemodialysed hypertensive patients, both endothelium-dependent and -independent vasodilation are impaired. Markers of endothelial damage are elevated referring the progression of vascular disease. PMID- 16468614 TI - [The role of MR angiography in predicting operative results of microvascular decompression in patients with trigeminal neuralgia]. AB - INTRODUCTION AND OBJECTIVES: The results of medium-term outcome of microvascular decompression (MVD) for trigeminal neuralgia are presented. The authors compare the preoperative 3-dimension magnetic resonance angiography (MRA) results with the surgical findings during MVD. Information, provided by MRA, are evaluated regarding to the prognostic significance in typical TN, atypical TN and persistent idiopathic facial pain (PIFP). The significance of clinical symptoms and the type of neurovascular compression (NC) are investigated in respect of the postoperative success and recurrent symptoms. METHODS AND RESULTS: MRA was performed in 310 consecutive patients with TN and PIFP. The MRA image was positive in 179 (58%) of the 310 cases. 68.2% of the typical TN group, 49.2% of the atypical TN group and 3.2% of the PIFP group were positive. MVD was performed in 116 of the MRA positive cases. Four years following the MVD, 69% of the patients gave an excellent and 23% a good result. The surgical findings corresponded with the MRA images. NC could be ruled out in the background of PIFP. The rate of recurrent symptoms following MVD is 21% in the typical TN group while it is 41% in the atypical TN group. The pure venous compression showed 57% pain-recurrence rate following MVD. CONCLUSIONS: The clinical symptoms and preoperative MRA carry considerable information, which can predict the outcome of the MVD and the rate of recurrent symptoms. Atypical TN and venous compression are bad prognostic factors. PMID- 16468615 TI - [Juvenile haemochromatosis presenting as intractable congestive heart failure]. AB - Juvenile haemochromatosis is an autosomal, recessive inherited iron metabolism disorder. The rapid deterioration and malignant prognosis differentiate juvenile haemochromatosis from hereditary haemochromatosis. The authors summarize the history of a 25 year old man, who worked in Hungary as a guest worker living in Romania. No significant illness has occurred in his previous history. The abdominal pain was his first symptom and he was treated in different institutions, where cholecystitis, alcoholic hepatic disease, hepatic cirrhosis were considered as a cause of his symptoms. Some weeks later atrial tachycardia, and congestive heart failure were observed and he was sent to our Cardiology Department. The echocardiography revealed diffuse hypokinesis, serious systolic dysfunction (ejection fraction: 21%), grade II mitral and tricuspid insufficiency with pulmonary hypertension. Considering the rapid deterioration of his cardiac function, myocarditis was suspected. Myocardial biopsy and coronary arteriography were performed. Coronary arteries were normal. Ventricular fibrillation occurred during coronary arteriography. Myocardial biopsy revealed juvenile haemochromatosis. Special laboratory examinations (transferrin saturation) were made after biopsy, that also confirmed the diagnosis of juvenile haemochromatosis. Cardiac transplantation was planned. Some days after the diagnosis was made the patient died of cardiogenic shock and intractable heart failure. Autopsy revealed hypogonadism and serious haemochromatosis in different parenchymal organs. Juvenile haemochromatosis should be considered in every young patient with congestive heart failure of unknown etiology. PMID- 16468616 TI - Kernel matched subspace detectors for hyperspectral target detection. AB - In this paper, we present a kernel realization of a matched subspace detector (MSD) that is based on a subspace mixture model defined in a high-dimensional feature space associated with a kernel function. The linear subspace mixture model for the MSD is first reformulated in a high-dimensional feature space and then the corresponding expression for the generalized likelihood ratio test (GLRT) is obtained for this model. The subspace mixture model in the feature space and its corresponding GLRT expression are equivalent to a nonlinear subspace mixture model with a corresponding nonlinear GLRT expression in the original input space. In order to address the intractability of the GLRT in the feature space, we kernelize the GLRT expression using the kernel eigenvector representations as well as the kernel trick where dot products in the feature space are implicitly computed by kernels. The proposed kernel-based nonlinear detector, so-called kernel matched subspace detector (KMSD), is applied to several hyperspectral images to detect targets of interest. KMSD showed superior detection performance over the conventional MSD when tested on several synthetic data and real hyperspectral imagery. PMID- 16468617 TI - Restoring warped document images through 3D shape modeling. AB - Scanning a document page from a thick bound volume often results in two kinds of distortions in the scanned image, i.e., shade along the "spine" of the book and warping in the shade area. In this paper, we propose an efficient restoration method based on the discovery of the 3D shape of a book surface from the shading information in a scanned document image. From a technical point of view, this shape from shading (SFS) problem in real-world environments is characterized by 1) a proximal and moving light source, 2) Lambertian reflection, 3) nonuniform albedo distribution, and 4) document skew. Taking all these factors into account, we first build practical models (consisting of a 3D geometric model and a 3D optical model) for the practical scanning conditions to reconstruct the 3D shape of the book surface. We next restore the scanned document image using this shape based on deshading and dewarping models. Finally, we evaluate the restoration results by comparing our estimated surface shape with the real shape as well as the OCR performance on original and restored document images. The results show that the geometric and photometric distortions are mostly removed and the OCR results are improved markedly. PMID- 16468618 TI - Channel smoothing: efficient robust smoothing of low-level signal features. AB - In this paper, we present a new and efficient method to implement robust smoothing of low-level signal features: B-spline channel smoothing. This method consists of three steps: encoding of the signal features into channels, averaging of the channels, and decoding of the channels. We show that linear smoothing of channels is equivalent to robust smoothing of the signal features if we make use of quadratic B-splines to generate the channels. The linear decoding from B spline channels allows the derivation of a robust error norm, which is very similar to Tukey's biweight error norm. We compare channel smoothing with three other robust smoothing techniques: nonlinear diffusion, bilateral filtering, and mean-shift filtering, both theoretically and on a 2D orientation-data smoothing task. Channel smoothing is found to be superior in four respects: It has a lower computational complexity, it is easy to implement, it chooses the global minimum error instead of the nearest local minimum, and it can also be used on nonlinear spaces, such as orientation space. PMID- 16468619 TI - A linear feature extraction for multiclass classification problems based on class mean and covariance discriminant information. AB - A parametric linear feature extraction method is proposed for multiclass classification. The skeleton of the proposed method consists of two types of schemes that are complementary to each other with regard to the discriminant information used. The approximate pairwise accuracy criterion (aPAC) and the common-mean feature extraction (CMFE) are chosen to exploit the discriminant information about class mean and about class covariance, respectively. Choosing aPAC rather than the linear discriminant analysis (LDA) can also resolve the problem of overemphasized large distances introduced by LDA, while maintaining other decent properties of LDA. To alleviate the suboptimum problem caused by a direct cascading of the two different types of schemes, there should be a mechanism for sorting and merging features based on their effectiveness. Usage of a sample-based classification error estimation for evaluation of effectiveness of features usually costs a lot of computational time. Therefore, we develop a fast spanning-tree-based parametric classification accuracy estimator as an intermediary for the aPAC and CMFE combination. The entire framework is parametric-based. This avoids paying a costly price in computation, which normally happens to the sample-based approach. Our experiments have shown that the proposed method can achieve a satisfactory performance on real data as well as simulated data. PMID- 16468620 TI - Data driven image models through continuous joint alignment. AB - This paper presents a family of techniques that we call congealing for modeling image classes from data. The idea is to start with a set of images and make them appear as similar as possible by removing variability along the known axes of variation. This technique can be used to eliminate "nuisance" variables such as affine deformations from handwritten digits or unwanted bias fields from magnetic resonance images. In addition to separating and modeling the latent images-i.e., the images without the nuisance variables-we can model the nuisance variables themselves, leading to factorized generative image models. When nuisance variable distributions are shared between classes, one can share the knowledge learned in one task with another task, leading to efficient learning. We demonstrate this process by building a handwritten digit classifier from just a single example of each class. In addition to applications in handwritten character recognition, we describe in detail the application of bias removal from magnetic resonance images. Unlike previous methods, we use a separate, nonparametric model for the intensity values at each pixel. This allows us to leverage the data from the MR images of different patients to remove bias from each other. Only very weak assumptions are made about the distributions of intensity values in the images. In addition to the digit and MR applications, we discuss a number of other uses of congealing and describe experiments about the robustness and consistency of the method. PMID- 16468621 TI - Attention-based dynamic visual search using inner-scene similarity: algorithms and bounds. AB - A visual search is required when applying a recognition process on a scene containing multiple objects. In such cases, we would like to avoid an exhaustive sequential search. This work proposes a dynamic visual search framework based mainly on innerscene similarity. Given a number of candidates (e.g., subimages), we hypothesize is that more visually similar candidates are more likely to have the same identity. We use this assumption for determining the order of attention. Both deterministic and stochastic approaches, relying on this hypothesis, are considered. Under the deterministic approach, we suggest a measure similar to Kolmogorov's epsilon-covering that quantifies the difficulty of a search task. We show that this measure bounds the performance of all search algorithms and suggest a simple algorithm that meets this bound. Under the stochastic approach, we model the identity of the candidates as a set of correlated random variables and derive a search procedure based on linear estimation. Several experiments are presented in which the statistical characteristics, search algorithm, and bound are evaluated and verified. PMID- 16468622 TI - Global segmentation and curvature analysis of volumetric data sets using trivariate B-spline functions. AB - This paper presents a method to globally segment volumetric images into regions that contain convex or concave (elliptic) iso-surfaces, planar or cylindrical (parabolic) iso-surfaces, and volumetric regions with saddle-like (hyperbolic) iso-surfaces, regardless of the value of the iso-surface level. The proposed scheme relies on a novel approach to globally compute, bound, and analyze the Gaussian and mean curvatures of an entire volumetric data set, using a trivariate B-spline volumetric representation. This scheme derives a new differential scalar field for a given volumetric scalar field, which could easily be adapted to other differential properties. Moreover, this scheme can set the basis for more precise and accurate segmentation of data sets targeting the identification of primitive parts. Since the proposed scheme employs piecewise continuous functions, it is precise and insensitive to aliasing. PMID- 16468623 TI - A dynamic conditional random field model for foreground and shadow segmentation. AB - This paper proposes a dynamic conditional random field (DCRF) model for foreground object and moving shadow segmentation in indoor video scenes. Given an image sequence, temporal dependencies of consecutive segmentation fields and spatial dependencies within each segmentation field are unified by a dynamic probabilistic framework based on the conditional random field (CRF). An efficient approximate filtering algorithm is derived for the DCRF model to recursively estimate the segmentation field from the history of observed images. The foreground and shadow segmentation method integrates both intensity and gradient features. Moreover, models of background, shadow, and gradient information are updated adaptively for nonstationary background processes. Experimental results show that the proposed approach can accurately detect moving objects and their cast shadows even in monocular grayscale video sequences. PMID- 16468624 TI - Stereo matching with linear superposition of layers. AB - In this paper, we address stereo matching in the presence of a class of non Lambertian effects, where image formation can be modeled as the additive superposition of layers at different depths. The presence of such effects makes it impossible for traditional stereo vision algorithms to recover depths using direct color matching-based methods. We develop several techniques to estimate both depths and colors of the component layers. Depth hypotheses are enumerated in pairs, one from each layer, in a nested plane sweep. For each pair of depth hypotheses, matching is accomplished using spatial-temporal differencing. We then use graph cut optimization to solve for the depths of both layers. This is followed by an iterative color update algorithm which we proved to be convergent. Our algorithm recovers depth and color estimates for both synthetic and real image sequences. PMID- 16468626 TI - Individual recognition using gait energy image. AB - In this paper, we propose a new spatio-temporal gait representation, called Gait Energy Image (GEI), to characterize human walking properties for individual recognition by gait. To address the problem of the lack of training templates, we also propose a novel approach for human recognition by combining statistical gait features from real and synthetic templates. We directly compute the real templates from training silhouette sequences, while we generate the synthetic templates from training sequences by simulating silhouette distortion. We use a statistical approach for learning effective features from real and synthetic templates. We compare the proposed GEI-based gait recognition approach with other gait recognition approaches on USF HumanID Database. Experimental results show that the proposed GEI is an effective and efficient gait representation for individual recognition, and the proposed approach achieves highly competitive performance with respect to the published gait recognition approaches. PMID- 16468625 TI - Robust structure and motion from outlines of smooth curved surfaces. AB - This paper addresses the problem of estimating the motion of a camera as it observes the outline (or apparent contour) of a solid bounded by a smooth surface in successive image frames. In this context, the surface points that project onto the outline of an object depend on the viewpoint and the only true correspondences between two outlines of the same object are the projections of frontier points where the viewing rays intersect in the tangent plane of the surface. In turn, the epipolar geometry is easily estimated once these correspondences have been identified. Given the apparent contours detected in an image sequence, a robust procedure based on RANSAC and a voting strategy is proposed to simultaneously estimate the camera configurations and a consistent set of frontier point projections by enforcing the redundancy of multiview epipolar geometry. The proposed approach is, in principle, applicable to orthographic, weak-perspective, and affine projection models. Experiments with nine real image sequences are presented for the orthographic projection case, including a quantitative comparison with the ground-truth data for the six data sets for which the latter information is available. Sample visual hulls have been computed from all image sequences for qualitative evaluation. PMID- 16468627 TI - Wavelet approximation-based affine invariant shape representation functions. AB - In this paper, new wavelet-based affine invariant functions for shape representation are presented. Unlike the previous representation functions, only the approximation coefficients are used to obtain the proposed functions. One of the derived functions is computed by applying a single wavelet transform; the other function is calculated by applying two different wavelet transforms with two different wavelet families. One drawback of the previously derived detail based invariant representation functions is that they are sensitive to noise at the finer scale levels, which limits the number of scale levels that can be used. The experimental results in this paper demonstrate that the proposed functions are more stable and less sensitive to noise than the detail-based functions. PMID- 16468628 TI - Implicit meshes for surface reconstruction. AB - Deformable 3D models can be represented either as traditional explicit surfaces, such as triangulated meshes, or as implicit surfaces. Explicit surfaces are widely accepted because they are simple to deform and render, but fitting them involves minimizing a nondifferentiable distance function. By contrast, implicit surfaces allow fitting by minimizing a differentiable algebraic distance, but are harder to meaningfully deform and render. Here, we propose a method that combines the strength of both approaches. It relies on a technique that can turn a completely arbitrary triangulated mesh, such as one taken from the Web, into an implicit surface that closely approximates it and can deform in tandem with it. This allows both automated algorithms to take advantage of the attractive properties of implicit surfaces for fitting purposes and people to use standard deformation tools they feel comfortable for interaction and animation purposes. We demonstrate the applicability of our technique to modeling the human upper body, including face, neck, shoulders, and ears, from noisy stereo and silhouette data. PMID- 16468629 TI - Expression of extracellular domain of human vascular endothelial growth factor receptor-2 in Pichia pastoris. AB - OBJECTIVE: To investigate the feasibility of high expressing extracellular domain of human vascular endothelial growth factor receptor-2 (heVEGFR-2) with eukaryotic protein structure in Pichia pastoris. METHODS: We used PCR to amplify the DNA fragment encoding heVEGFR-2 from pORF-heVEGFR-2. The recombinant Pichia pastoris secretory expression vector(pPICZalphaA-heVEGFR-2)was constructed and transferred into Pichia pastoris X-33 by electroporation. The high expression transformnants were identified through its drug-resistant phenotype and methanol induction. RESULTS: As indicated by SDS-PAGE, the recombinant heVEGFR-2 protein with a molecular weight (MW) approximately 108 kDa, which reached 80 mg/L in the mass concentration, comprised 45% of the total expressed secreted proteins from Pichia pastoris X-33. The section of heVEGFR-2 had a MW approximately 106 kDa. The results of western blot analysis demonstrated that this protein could be specifically recognized by the rat monoclonal antibody against mouse VEGFR-2 (rat McAb against mVEGFR-2). CONCLUSION: The heVEGFR-2 with eukaryotic protein structure can get a high expression in Pichia pastoris. PMID- 16468630 TI - Promoter activity of SARS coronavirus 5' UTR sequence in eukaryotic cells. AB - OBJECTIVES: To investigate 5'UTR sequence in different SARS-CoV isolates, to identify the secondary structure, and to test the promoter activity of the cDNA sequence corresponding to SARS-CoV 5'UTR in eukaryotic cells. METHODS: 101 SARS CoV 5'UTR were aligned. One typical sequence containing full 264 nt was then subjected to be predicted its secondary structure. The pGL3-5'UTR and pGL3-a 5'UTR were constructed by substitution of SV40 promoter with SARS-CoV 5'UTR cDNA or its antisense sequence. Then the recombinant plasmids were transfected into HepG2 cells and the luciferase activities were detected. A set of deletion mutant plasmids, of which pGL3-5' UTR-1, pGL3-5' UTR-2, pGL3-5'UTR-3 and pGL3-5'UTR-4 are with 3, 2, 1, and 0 residual stem-loops of 3' termini respectively,were constructed from pGL3-5'UTR and were transfected into HepG2 cells to express reporter gene luc+, with pGL3-5'UTR containing full sequence as control. The luciferase activities expressed by the plasmids were measured. And then the total RNA of the transfected cells was extracted. Subsequently, by 5' Rapid Amplication of cDNA Ends (5'RACE), the PCR product was sequenced. The luciferase expressed by pGL3-5'UTR in various cells, the lung carcinoma cell line A549, hepatoma cell line HepG2, kidney cell Vero E6, cervical cancer cell line HeLa and human umbilical vein endothelial cell line ECV304 were measured and compared with each other. RESULTS: The full sequence of the SARS-CoV 5' UTR is a 264nt, and 18 deletion mutants were found. Totally, 5 site substitutions were found in 101 5'UTR sequences. The SARS-CoV 5'UTR RNA folded to form a stable secondary structure containing four stem-loop domains. The biggest and most complex one is the stem-loop II appearing a pseudoknot. Comparing with pGL3-a-5'UTR, pGL3-5'UTR expressed luciferase obviously. Both pGL3-5'UTR containing full sequence and pGL3 5'UTR-1 containing three stem-loops of 3' termini expressed the luciferase well. However, when lost stem-loop I and II , the pGL3-5'UTR-2, pGL3-5'UTR-3 and pGL3 5'UTR-4 almost didn't express luciferase. The 56th nucleotide of SARS-CoV 5'UTR was found to be the initiation site for transcription. Transfected with expression luciferase plasmid pGL3-5' UTR in which SARS-CoV 5' UTR acts as the promoter, the luciferase could express in five cell lines in different degrees. Ranked by the luciferase activity from the highest to the lowest, the order is A549, HepG2, ECV304, HeLa and Vero E6. CONCLUSIONS: A: The 5'UTR sequences of different SARS-CoV isolates are relatively conserved, and a full sequence would form a secondary structure containing four stem-loop domains. B: The cDNA sequence corresponding to SARS-CoV 5'UTR possessed a promoter activity in eukaryotic cells. C: The promoter domain of the SARS-CoV 5'UTR contains both stem loop I and II. D: The 56th nucleotide and its down stream TRS of SARS-CoV 5'UTR plays a key role in regulating transcription. E: Cells sourced from various tissues can provide efficient accessory factors for SARS-CoV 5'UTR sequence that acts as a promoter, and the lung-sourced cells may be the most suitable. PMID- 16468631 TI - [Studies of the possible mechanisms for protective effects of recombinant human growth hormone on intestinal mucosa barrier in rat sepsis]. AB - OBJECTIVE: To investigate the protective effects of recombinant human growth hormone (rhGH) on intestinal mucosal barrier in rat sepsis and explore its possible mechanisms. METHODS: E. coli was injected intraperitoneally to produce rats sepsis models. Forty-two female SD rats were randomly divided into the control group (group C), sepsis group (group S) and treatment group (group T). Group S and group T were further divided into 1 d and 3 d subgroups (T1d,T3d, Sld, S3d), respectively. The expression of IGF-1 mRNA in liver, expression of Bcl 2 protein in intestine, bacteria translocation, the levels of growth hormone(GH) and insulin-like growth factor-1 (IGF-1) in plasma, and the histological appearance of intestine were determined dynamically by means of RT-PCR, radioimmunoassay, immunohistochemical staining and other corresponding methods, respectively. RESULTS: (1) rhGH could significantly attenuate intestinal mucosal injuries and ameliorate bacteria translocation on sepsis rats. (2) The levels of Bcl-2 protein expression in intestine in group T (T1d:2441 +/- 117; T3d: 3628 +/- 235) were obviously higher than those of group S (S1d: 321 +/- 36; S3d: 1873 +/- 57) (P < 0.01). (3) The plasma levels of GH in group T (T1d: 1.28 +/- 0.24 microg/L; T3d: 2.14 +/- 0.48 microg/L) increased markedly than those of group S (S1d: 0.74 -/+ 0.12 microg/L; S3d: 0.60 +/- 0.18 microg/L) (P < 0.01). (4) The plasma levels of IGF-1 in group T (Tld: 168.94 +/- 65.67 microg/L; T3d: 201.56 +/ 64.98 microg/L) elevated significantly than those of group S (Sld: 116.72 +/- 13.96 microg/L; S3d:107.50 +/- 23.53 microg/L) (P < 0.05). (5) The levels of liver IGF-1 mRNA in group T (Tld: 0.98 +/- 0.20; T3d: 1.76 +/- 0.17) were significantly higher than those in group S (S1d: 0.38 +/- 0.09; S3d: 0.46 +/- 0.10) (P < 0.01). CONCLUSION: rhGH conferred protective efficacy in maintaining the integrity of intestinal mucosal barrier against sepsis in rat. The possible mechanisms might involve the rhGH-diminished apoptosis of intestinal mucosa cells and the rhGH-maintained intestinal mucosa barrier via the roles of GH and IGF-1. PMID- 16468632 TI - [Inhibition of HPV16 E6 oncogene in cervical cancer by RNA interference]. AB - OBJECTIVE: The efficiency of HPV16 E6 gene silenced by RNA interference in vitro and in vivo was assessed. METHODS: The specific siRNA of HPV16 E6 was designed and transfected into CaSki cells by liposome. Cell apoptotic rates and the changes in HPV16 E6 mRNA and protein before and after transfection were measured. Cervical cancer nude mice models were set up, siRNA was injected directly into subcutaneous tumor. The function of siRNA was evaluated by the changes in tumor volume, HPV16 E6 protein expression and apoptosis of tumor cells. RESULTS: In vitro research, the cell apoptotic rates were 7.7%, 11.8%, 37.4% and 12.6% respectively at 24 h, 48 h, 5th day and 9th day after transfection. The HPV16 E6 mRNA was reduced by 77%, 83%, 59% and 41% at 24 h, 48 h, 5th day and 9th day after transfection. The inhibition rates of E6 protein measured by Flow cytometry were 79.7%, 80.4%, 71.3% and 57.4% at 24 h, 48 h, 5th day and 9th day after transfection, which were confirmed by the results of Western blot. In vivo research, E6 siRNA administration groups had great power in inhibiting tumor growth, restraining E6 protein expression, increasing tumor necrosis and apoptosis. The result of repeated injections of siRNA was better than that of single injection. CONCLUSION: RNA interference with HPV16 E6 is specific and highly efficient in vitro and in vivo. PMID- 16468633 TI - [Inducing apoptosis of HeLa cell lines by transfection of c-myc antisense oligonucleotides]. AB - OBJECTIVE: To investigate the effect on apoptosis of HeLa cells by transfecting c myc antisense oligonucleotides and seek a new method to enhance radiosensitivity of tumor cells. METHODS: Immunohistochemistry staining and RT-PCR were conducted to identify the effect of ASODN. Giemsa staining, flow cytometry (FCM) and DNA ladder were performed to detect the apoptosis of HeLa cells. RESULTS: After transfection of ASODN (c-myc), the levels of both c-myc mRNA and C-MYC protein expression were reduced (P < 0.05). Giemsa staining demonstrated apoptosis-bodies on the c-myc ASODN-transfected cells. DNA ladder presented the typical ladder of apoptosis. The apoptosis rate of HeLa cells transfected with c-myc ASODN was (10.29 +/- 0.66)%, transfected c-myc ASODN combined irradiation was (16.83 +/- 0.57)%, which were much higher than that of control group (1.79 +/- 0.19)% (P < 0.05). CONCLUSION: The c-myc ASODN could effectively down-regulate the expression of c-myc; furthermore, transfection of c-myc ASODN could more effectively induce apoptosis of HeLa cells which subsequently enhanced radiosensitivity of HeLa cells. PMID- 16468634 TI - [Effects of nonylphenol on PCNA expression and aromatase expression in reproductive system of F1 generation]. AB - OBJECTIVE: To elucidate the mechanism of the toxicity of nonylphenol (NP) to reproductive development by analyzing the PCNA and Aromatase expression of the F1 generation male reproductive system in perinatal period. METHODS: Nonylphenol was administrated to pregnant rats at doses of 50, 100 and 200 mg/kg, respectively, for 7 to 20 days of pregnancy. The rats were sacrificed at 20 days of pregnancy and 2 days after parturition. Their serum levels of estradiol and nonylphenol were determined. The histopathological examination and immune histochemical analysis on testis and epididymis were also performed. RESULTS: Significantly higher serum estradiol and nonylphenol levels were found in the pregnant rats treated with nonylphenol. Immune histochemical analysis showed that the PCNA expression levels of testis tissues of F1 generation rats, both 20-day-old fetus and 2-day-old neonatal rats, in 100 mg/kg and 200 mg/kg experiment groups were much lower than those in the control group. Both PCNA expression of epididymis and Aromatase expression of testis of the F1 generation rats after 2 days of birth in 200 mg/kg group were also lower than those in the control group. CONCLUSION: Nonylphenol can impair the development of testis and epididymis of the F1 generation in perinatal period. PMID- 16468635 TI - [Expression of Cyt C in brain of rats poisoned by tetramine]. AB - OBJECTIVE: To investigate the evidence for the judgement on legal cases of intoxication by tetramine. METHODS: Fifty rats given tetramine by lavage were sacrificed at different time after intoxication. The expression of Cyt C was detected by immunohistochemistry, and the reasults were assessed by an image analysis system. RESULTS: There was a slight expression of Cyt C in control group. In intoxication group the expression of Cyt C increased after the rats were given tetramine for 30 minutes; it reached the peak at 24 hours after intoxication and kept up the level for 3 days, then it began to decrease and returned to the control level in 10 days. CONCLUSION: The regular change of Cyt C expression might be a useful sign for assessment in legal cases of intoxication by tetramine. PMID- 16468636 TI - [Study of rat's p53 gene damage and organ specificity induced by benzidine]. AB - OBJECTIVE: Studying the main target organs and the genetic toxicity mechanism of benzidine. METHODS: SD rats were given benzidine i.p. injection. DNA was extracted from rat's liver, kidney, lung and bladders. Then ss probes RDPCR was used to detect the damaged DNA of exon 7 of p53 gene. RESULTS: Hybridization bands were found in liver, bladder and lung tissues after ss probes RDPCR, while no hybridization bands were found in kidney tissues. CONCLUSION: The result indicates that benzidine can cause the DNA lesions of exon 7 of p53 gene and its major target organs are liver, bladder and lung. The toxicity mechanism of benzidine is probably related to p53 gene damage. PMID- 16468638 TI - [The effect of serum rich in platelet-released growth factors on proliferation of rat osteoblast in vitro]. AB - OBJECTIVE: To study the effect of serum rich in platlelet-released growth factors on the biological functions of rat osteoblast. METHODS: Rat osteoblasts were isolated from parietal bone of fetal Spraque-Dawley (SD) rat. Platelet-rich plasma(PRP) and platelet-poor plasma (PPP) obtained from healthy SD rat were activated by thrombin to get serum rich in growth factors (SRGF) or serum poor in growth factors (SPGF). The cells were treated with 1.25%, 2.5%, 5% SRGF or SPGF, and cellular mitogenic activity was evaluated by thiazoly blue (MTT) colorimetric assay. Results When compared with SPGF at the same concentration, SRGF promoted the proliferation of rat osteoblast significantly (P = 0.002 - 0.004), and the proliferation seemed to correlate with the concentration of SRGF (Spearman's correlation coefficient, r(s) = 0.834). CONCLUSION: SRGF is capable of upregulating the proliferation of rat osteoblast. PMID- 16468637 TI - [On the role of liver-enriched transcription factors in regulating HBV transcription and replication]. AB - OBJECTIVE: To investigate the effects of various liver-enriched transcription factors in regulating HBV transcription and replication, and to explore their potential roles in HBV hepatotropism. METHODS: The replication-competent HBV recombinant plasmid pHBV4.1 plus different liver-enriched transcription factor (HNF1, HNF3, HNF4, HNF6, C/EBP and RXRa/PPARa) expression plasmids were co transfected into nonhepatic cell lines (NIH3T3, HeLa, 293T, SW1353, CV-1 and COS1). The transcription levels of 3.5 kb, 2.4/2.1 kb and 0.7 kb HBV RNA were analyzed by Northern blot hybridization, and the level of HBV DNA replication intermediates was detected by Southern blot hybridization analysis. RESULTS: In the absence of co-transfected liver enriched transcription factor expression vectors, the 3.5 kb HBV RNA is not transcribed and HBV DNA replication is not detected after transfecting of NIH 3T3 cells with pHBV4.1. Expression of the liver-enriched transcription factor HNF4 or RXRalpha/PPARalpha, stimulates the transcription of 3.5 kb HBV RNA and the replication of HBV DNA. In contrast, expression of HNF1, HNF3, HNF6 and C/EBP does not stimulate the transcription of 3.5 kb HBV RNA and therefore does not activate viral replication. HNF4 and RXRalpha/PPARalpha were also shown to activate the transcription of 3.5 kb HBV RNA and viral replication in divers cell types including HeLa, 293T, SW1353, CV-1 and COS1 cells. Mutation of the proximal nucleocapsid HNF4 binding site results in a greatly decreased level of HNF4 or RXRalpha/PPARalpha dependent HBV replication. CONCLUSION: This study demonstrated that the liver-enriched transcription factors HNF4 and RXRa/PPARa can support HBV transcription and replication in nonhepatic cells, indicating that liver-specific gene transcription is one of the determinants of HBV hepatotropism. PMID- 16468639 TI - [Experimental study on the proliferation and function of osteoblast cell induced by pBLAST49-mVEGF gene transfection]. AB - OBJECTIVE: To assess the effects of extra VEGF gene expression on the proliferation of osteoblast cell. METHODS: The extra VEGF gene was induced into osteoblast cell. Then the expression of extra VEGF gene was detected, and its effects on the cell proliferation curve, cell cycle, on the synthesis of type I collagen and on the secretion of osteocalcium of osteoblast cell were investigated. RESULTS: At the first two days of culture, the cell number of pBLAST49-mVEGF gene transfer group was larger than that of the control group, yet no statistically significant difference was seen between the two groups (P > 0.05); however, significant difference was observed from the 3rd day of culture (P < 0.05). The (G2/M+S)% of the 1-5 generation osteoblast cell in pBLAST49-mVEGF gene transfer group was higher than that of the control group, there was significant difference between them (P < 0.05). The concentration of osteocalcium and expression of type I collagen of the 1-5 generation osteoblast cell in pBLAST49-mVEGF gene transfer group were significantly higher than those of the control group (P < 0.05). CONCLUSION: pBLAST49-mVEGF gene transfer can improve the proliferation and function of osteoblast cell. PMID- 16468640 TI - [Role of Ph-SA in enhancing bactericidal activity of beta-lactam antibiotics]. AB - OBJECTIVE: To identify the role of Ph-SA in inhibiting beta-lactamase secretion of S. aureus and in enhancing the bactericidal activity of antibiotics against antibiotic resistant bacteria. METHODS: To determine the effects of Ph-SA on beta lactamase product of bacteria and on bactericidal activity of penicillin, the growth rate and lactamase activity of penicillin-resistant S. aureus cells in exponential growth were detected after treatment with 1.43 X 10(-4) micromol/mL (10 microg/mL) Ph-SA (Group 1), 2.81 x 10(-2) micromol/mL (10 microg/mL) penicillin (Group 2), 0.715 x 10(-4) micromol/mL(5 microg/mL) Ph-SA + 1.40 x 10( 2) micromol/mL (5 microg/mL) penicillin (Group 3) respectively; and a control group was detected for comparison. The same method was used to detect whether Ph SA could enhance bactericidal activity of Cefazolin. Results It was revealed that 1.43 x 10(-4) micromol/mL (10 microg/mL) Ph-SA could inhibit beta-lactamase activity significantly, whereas no marked bactericidal activity was detected. The growth rate of bacteria cells treated with 0.715 x 10(-4) micromol/mL Ph-SA + 1.40 x 10(-2) mol/mL penicillin and that of bacteria cells treated with 0. 715 x 10(-4) micromol/mL (5 microg/mL) Ph-SA + 1.05 x 10(-2) micromol/mL (5 microg/mL) Cefazolin were the same as those treated by 2.81 x 10(-)2 micromol/mL (10 microg/mL)penicillin and 2.10 x 10(-2) micromol/mL (10 microg/mL) Cefazolin respectively, while the beta-lactamase activities of both groups were lower than control. CONCLUSION: Ph-SA may be of value in enhancing the bactericidal activity of antibiotics and in alleviating the resistance of bacteria by inhibition of beta-lactamase activity. PMID- 16468641 TI - [Relief effect of beta-galactosidase genetically engineered lactococcus lactis on the cell toxicity caused by lactose]. AB - OBJECTIVE: To assess the relief effect of beta-galactosidase genetically engineered Lactococcus lactis on the cell toxicity caused by lactose in vitro. METHODS: An in vitro toxic Caco-2 cell model caused by lactose was established to evaluate the relief effect of beta-galactosidase genetically engineered Lactococcus lactis. Cell morphological parameters and proliferation activity parameter were used. RESULTS: The in vitro toxic Caco-2 cell model caused by lactose was successfully established; the genetically engineered Lactococcus lactis constructed in the authors' laboratory could enable the Caco-2 cell to have normal appearance with the presence of lactose and could improve the proliferation activity with the presence of high concentration of lactose (P < 0.01). CONCLUSION: The beta-galactosidase genetically engineered Lactococcus lactis has significant relief effect on the cell toxicity caused by lactose in vitro, which lays a foundation for food-grade alternation of this bacterium. PMID- 16468642 TI - [Effects of Tanshinone IIA on procoagulant activity of human ECV304 cell line induced by NB4 cells]. AB - OBJECTIVE: To investigate the effects of Tanshinone IIA (Tan IIA) on procoagulant activity (PCA) of human ECV304 cells induced by acute promyelocytic leukemia cell line NB4 cells. METHODS: ECV304 monolayers were respectively incubated for different hours at 37 degrees C in the conditioned media (CM) of NB4 cells treated with 0.5 microg/mL Tan IIA(Tan IIA-NB4-CM), 0.3 microg/mL all-trans retinoidic acid (ATRA)(ATRA-NB4-CM), DMSO(DMSO-NB4-CM) or the RPMI1640 medium. ECV304 lysates were tested for PCA using the one-stage clotting assay as well as for tissue factor activity (TF: Act) using the chromogenic substrate assay; ECV304 cell monolayers were incubated for different hours at 37 degrees C in a medium system including 0.5 microg/mL Tan IIA and Tan IIA-NB4-CM, and the ECV304 cell lysates were tested for PCA in the same way as above. Also they were controlled by 0.3 microg/mL ATRA, DMSO or RPMI1640 medium. RESULTS: (1) The conditioned mediums from 0. 5 microg/mL Tan IIA that treated NB4 cells for 24, 72 and 120 hours respectively could elevate PCA of ECV cells, and this capability developed with the time of reaction. ATRA did the same as Tan IIA (P > 0.05). (2) 0.5 microg/mL Tan IIA down-regulated the PCA of ECV304 cells induced by Tan IIA NB4-CM, and the inhibitory effects increased with time, reaching the highest at 120 hours. (3) Tan IIA120 h-NB4-CM up-regulated TF:Act of ECV304 cells, and the effect increased with time. (4) 0. 5 microg/mL Tan IIA down-regulated PCA and TF: Act of ECV304 cells induced by Tan IIA-NB4-CM, and the inhibitory effect increased with time; simultaneously, the test was controlled with 0.3 microg/mL ATRA, the effects on PCA and TF: Act were not significantly different (P > 0.05). CONCLUSION: Tan IIA-NB4-CM can increase the levels of PCA and TF: Act of ECV304 cells through some unidentified factor; however, Tan IIA can obviously decrease the PCA and TF: Act levels of ECV304 cells induced by Tan IIA-NB4-CM. PMID- 16468643 TI - [The effects of 20(R)-Rg3 on lung carcinoma A549 cell line and endogenous VEGF secreted by tumor cells]. AB - OBJECTIVE: To assess the effect of 20(R)-Rg3 on the inhibition of angiogenesis of lung cancer and explore its role in the change of endogenous VEGF secreted by A549 tumor cells themselves. METHODS: A549 tumor cells were cultured with different concentrations of 20(R)-Rg3(10(-7) mol/L, 10(-6) mol/L, 10(-5) mol/L and 10(-4), mol/L), the VEGF contents were detected by ELASA method after 24 h, 48 h and 72 h. Cellular proliferation was detected by MTT method, and apoptosis was detected with flow cytometry. RESULTS: It was found that Rg3 had no significant inhibitory effect on the proliferation of A549 cells. The apoptosis rate was 29.8% after the cells being interfered with Rg3 at 3 x 10(-5) mol/L for 120 h. The levels of VEGF in the groups of Rg3 at 10(-5) mol/L and 10(-4) mol/L were lower than that in the contrast group (P < 0.05). The level of VEGF in the group of 10(-4) mol/L was lower than that in the group of 10(-7) mol/L and group of 10(-6) mol/L, and it was lower after 72 h of interference than that after 24 h and 48 h of interfernce. CONCLUSION: With the action of 20 (R)-Rg3, the level of VEGF secreted by A549 tumor cells themselves declined; the apoptosis of A549 tumor cells increased, especially at higher concentration and longer action time of Rg3. This might be one of the antiangiogenesis mechanisms of 20(R)-Rg3. PMID- 16468644 TI - [Effect of Huajiao volatile oil on Cu2+ -induced oxidative modification of low density lipoprotein in vitro]. AB - OBJECTIVE: To evaluate the effect of Huajiao volatile oil on copper mediated low density lipoprotein (LDL) oxidative modification. METHODS: LDL was isolated by ultracenfugation from normal human plasma. The extent of LDL peroxidation was measured in terms of thiobarbituric acid reactive substances (TBARS) expressed as malondialdehyde (MDA) equivalents. The electrophoresis mobility of LDL was determined as relative electrophoresis mobility (REM) on an agarose gel electrophoresis. RESULTS: The TBARS generation and the REM in Cu2+ -induced oxidation of LDL with Huajiao volatile oil at different concentrations were statistically different (P < 0.01). The Cu2+ -induced oxidation of LDL without addition of Huajiao volatile oil exhibited a lag phase of 1 h, a rapid increasing phase at 2 h-4 h, and a summit phase starting at 24 h. Then, with the addition of Huajiao volatile oil (40 mg/mL), the lag phase lasted 4 h, the rapid increasing phase was seen at 6 h-8 h, but the summit of TBARS was not on the decline. CONCLUSION: Huajiao volatile oil could protect LDL against Cu2+ -induced oxidative modification in vitro. PMID- 16468646 TI - [An experimental research of hyperthermic preconditioning on the spinal cord injury]. AB - OBJECTIVE: To assess the effects of hyperthermic preconditioning on spinal cord injury(SCI) in rats. METHODS: A total of 96 SD rats were divided into control(A), trauma(B) and hyperthermic preconditioning (C) group. SCI models were made by static compression. The animals were decapitated at 6 h, 12 h, 1 d, 2 d, 3 d, 1 w, 2 w and 3 w after injury. With in situ hybridization and immunohistochemistry, the expression of HSP70 after spinal cord injury was studied at the transcript level and translation level respectively. Neurological outcome was evaluated by the combined behavioral score (CBS). RESULTS: Neurological outcome in Group C was significantly higher than that in Group B (P < 0.05). In-situ hybridization and Immunohistochemistry analysis showed that expression of HSP70 in spinal cord elevated after hyperthermic preconditioning; furthermore, the level of HSP70 in Group C was significantly higher than that in Group B (P < 0.05). CONCLUSION: Hyperthermic preconditioning might improve neurological outcome after spinal cord injury in rats, and the protective mechanism in this connection might involve the induction of HSP70 synthesis in spinal cord. PMID- 16468645 TI - [Impact of Chai Qin Cheng Qi Decoction on cholinergic anti-inflammatory pathway in rats with severe acute pancreatitis]. AB - OBJECTIVE: To test the change of true choline esterase (TChE) and cholineacetyltransferase (ChAT) and their correlation with interleukin-6 (IL-6) and Tumor Necrosis Factor-alpha (TNF-alpha), and the impact of Chai Qin Cheng Qi Decoction (CQCQD) on cholinergic anti-inflammatory pathway in rats with severe acute pancreatitis (SAP). Methods Thirty SD rats were randomly divided into three groups: control group (pseudo-operated), SAP group and CQCQD-treated group, each with 10 rats. Blood samples were taken six hours after injection of testing agents for biochemical test, which included the test of amylase, TNF-alpha, IL-6, TChE, and ChAT. RESULTS: The rats in SAP group had higher levels of serum IL-6, TNF-alpha and TChE and lower levels of serum ChAT than those in control group (P < 0.05). The serum IL-6 was positively correlated with TChE (r = 0.95, P = 0.000) and negatively correlated with ChAT (r = -0.91, P = 0.000). The TNF-alpha was also positively correlated with TChE (r = 0.93, P = 0.000) and negatively correlated with ChAT (r = -0.95, P = 0.004). The rats in CQCQD-treated group had lower levels of serum IL-6, TNF-alpha and TChE and higher levels of serum ChAT than those in SAP group (P < 0.01). The increase of white blood cell, alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase was observed in the control group first and followed by the CQCQD-treated group and SAP group sequentially (P < 0.05). Conclusions Cholinergic anti-inflammatory pathway plays an important role in the pathological changes of SAP in rats. CQCQD can relieve the systemic inflammatory response syndrome and reduce the functional damage of organs through interference on the cholinergic antiinflammatory pathway. More studies are needed to reveal the mechanism of such impact. PMID- 16468647 TI - [Change of brainstem auditory evoked potential with different stimulating frequencies after closed diffuse brain injuries of rat]. AB - OBJECTIVE To investigate the change of peak latency (PL), interpeak latency (IPL) and peak latency ratio (PL ratio) of brainstem auditory evoked potential (BAEP) with different stimulating frequencies after closed diffuse brain injuries of rat,and provide experimental evidence for clinical diagnosis of traumatic brain injury in early stage and also for Forensic practice. METHODS: A rat model for closed diffuse brain injury was successfully constructed by hit the parietal bone with a spring-driven steel stick. The auditory brainstem response (ABR) evoked by clicking with low stimulating frequency of 10 Hz and high stimulating frequency of 50 Hz were respectively recorded at 6 h, 12 h, 2 d, 4 d after brain injury. The results of experimented group were compared with those of control group. RESULTS: The 2 d, 4 d V PL and the 6 h, 12 h, 2 d, 4 d III-V IPL of BAEP with 50 Hz stimulating frequency of model group became longer than those of control group (P < 0.05), but there was no significant difference observed between model and control group experimented with low stimulating frequency of 10 Hz (P > 0.05). The PL ratio of III/I, IV/I, V/I, IV/III and V/III with different stimulating frequencies had no significant postinjury changes. Conclusion The high stimulating frequency of BAEP is a reliable test for diagnosis of TBI in early period. Its sensitivity is superior to the low simulating frequency. PMID- 16468649 TI - [The analysis of T cell receptor Vbeta gene usage of cytolytic T-lymphocytes in coxsackievirus B3-induced myocarditis]. AB - OBJECTIVE: To analyze the T cell receptor (TCR) Vbeta gene usage of the cytolytic T-lymphocytes which result in immune-mediated myocyte injury in Coxsackievirus B3 induced myocarditis. METHODS: An experimental murine model with a myocarditic variant of Coxsackievirus B, type 3 has been developed with newborn Balb/c male mice. Mice were killed on day 7 after virus inoculated, which was the time when T cells show significant differentiation. With immunoadsorption, ACTLs, VCTLs, MCTLs are respectively isolated from mesenteric lymph nodes of Balb/c mice suffered from viral myocarditis. T-lymphocytes prepared from mesenteric lymph nodes of uninfected Balb/c mice served as control group. RT-PCR was performed with standard method. The Vbeta gene usage was analyzed by running the electophoresis of PCR products on agarose gel. RESULTS: T cells of control group expressed all Vbeta genes of 20 families. But the TCR VP gene usage of CTLs in experimental groups was markedly restricted. ACTL expressed predominantly Vbeta6, Vbeta8.1, Vbeta8.2, Vbeta8.3, MCTL expressed predominantly Vbeta5.1, Vbeta8.1, Vbeta8.2, Vbeta8.3 and VCTL expressed predominantly Vbeta7, Vbeta8.1, Vbeta8.2, Vbeta8.3. CONCLUSIONS: The TCR Vbeta gene usage of CTLs which result in immune mediated myocyte injury in Coxsackievirus B3-induced myocarditis is markedly restricted in cells stimulated by specific antigen. PMID- 16468648 TI - [Tumor cell targetability of folate receptor-mediated mitoxantrone albumin nanoparticles]. AB - OBJECTIVE: To study the tumor cell targetability of folate-conjugated mitoxantone loaded albumin nanoparticles (MTO-BSANP-folate). METHODS: Bovine albumin nanoparticles were prepared by desolvation method. The activated folic acid (N hydroxysuccinimide ester of folic acid) was conjuated to the surface of BSANP via the amino groups. The MTO-BSANP-folate was prepared by mixing folate-conjugated albumin nanoparticles with mitoxantrone and then cross-linked by glutaraldehyde. 3HTdR and flow cytometry were used to evaluate the targetability of MTO-BSANP folate. RESULTS: The encapsulation rate of folate-conjugated mitoxantrone albumin nanoparticles was (96.55 +/- 0.96)% and the drug loading was (9.66 +/- 0.10)%. The results of 3HTdR showed that the efficacy of MTO-BSANP-folate in killing SKOV3 cells was higher than that of MTO-BSANP-folate, and the results of flow cytometry showed that the apoptosis-promoting effect of MTO-BSANP-folate was 3.5 4.5 times higher than that of MTO-BSANP. CONCLUSION: MTO-BSANP-folate could be targeted, via folate receptor, to the tumor cells rich in folate receptors. PMID- 16468650 TI - [Analysis of the multi-amplified 5 STR loci and their allelic distribution in both Han populations in Chengdu City and Yunnan province]. AB - OBJECTIVE: To illuminate the multi-amplified 5 STR loci and their allelic distribution in Hans by means of STR-DNA typing with improved efficiency and decreased cost. METHODS: We have established an allelic ladder of D7S820, D13S317, D5S818, D3S1358 and Amelogenin loci via the cloning techniques. With this homemade allelic ladder, we established successfully a multiplexing polymerase chain reaction (PCR) method, followed by denaturing polyacrylamide gel electrophoresis (PAGE) and silver staining. DNA samples collected from 130 unrelated Han individuals in Yunnan and Chengdu were analyzed. The non overlapping of the allelic fragments of the five loci allowed the detection to be accomplished successfully. RESULTS: No difference of the genotyping results of the single locus amplification and multiplexing was observed. The genotype distributions of 4 STR were in accordance with the Hardy-Weinberg equilibrium. 7, 7, 8 and 8 alleles of D7S820, D13S317, D5S818, and D3S1358 loci were observed in Yunnan Han population, as well as 8, 7, 8 and 7 alleles in Chengdu Han population respectively. No significant difference in the allele distribution of these loci was seen between these two Han populations. CONCLUSION: This multiplexing system with home-made allelic ladder has a high combined discrimination power and exclusion power. It is a valuable tool in forensic science practice. PMID- 16468651 TI - [Adult-to-adult living donor liver transplantation]. AB - OBJECTIVE: To investigate the approach which could improve the survival rate of adult-to adult living donor liver transplantations, that right lobe liver grafts were performed by a modified technique. METHODS: During from March to June 2005, 13 patients underwent the living donor liver transplantation with right lobe grafts via the improved surgical techniques including the reconstruction of right hepatic vein; the tributaries of the middle hepatic vein were reconstructed by interpositioning a vein grafts; the anastomosis of the hepatic arteries and bile ducts. All cases underwent the direct anastomosis of right hepatic vein and IVC, of whom 5 cases were added with the reconstructions of right inferior hepatic vein, and other 5 cases added with reconstructing the tributaries of the middle hepatic vein by interpositing a vein graft to provide sufficient venous outflow. RESULTS: No donor suffered from severe complication or death. Four complications occurred in recipients who got the hepatic artery thrombosis (1 case), bile leakage (1 case), right subphrenic abscess (1 case) or pulmonary infection (1 case) respectively. The recipient with pulmonary infection died of MOF. The graft organ and recipient weight ratio (GRWR) were between 0.72% and 1.24%, that the ratios of 9 cases was < 1.0% and 2 cases < 0.8%, with no "small-for-size syndrome" occurred. CONCLUSIONS: The improved surgical techniques, especially the reconstruction of hepatic vein to provide sufficient venous outflow, can make the adult-to-adult living donor liver transplantation with right lobe liver grafts become a relatively safe operation and prevent the "small-for-size syndrome". PMID- 16468652 TI - [Solitary plasmacytoma: a clinicopathologic and immunophenotypic analysis of 43 cases]. AB - OBJECTIVE: To investigate the clinicopathologic and immunophenotypic features of solitary plasmacytoma and discuss the diagnosis and differential diagnosis of the tumor. METHODS: Clinicopathological study and follow up study were made on 43 cases of solitary plasmacytoma, including 24 cases of extramedullary plasmacytoma (EMP) and 19 cases of solitary plasmacytoma of bone (SPB). Immunohistochemical staining was performed by SP method using antibodies CD20, CD79a, PC, Ig kappa, Ig lamda, IgA, IgD, IgE, IgG and IgM. RESULTS: Rgarding EMP, the male/female ratio is 2.5 to 1, the age ranges from 3 to 70 years (mean 49). Clinically, the neoplasms of 22 cases were located in nasal cavity or nasosinus, 2 cases were located in lung and tonsil respectively. Rgarding SPB, the male/female ratio is 1:1, the age ranges from 36 to 72 years (mean 53). Clinically, the neoplasms of 8 cases were located in vertebrae, 3 cases in pelvis, 6 cases in clavicle, rib and femur respectively, 2 case in radius and tibia respectively. Histopathological examination revealed tumor cells diffuse infiltration with necrosis and ulceration; the well-differentiated tumor cells were similar to the normal plasmacyte in shape; the poorly-differentiated tumor resembled centroblasts; there were a few interstitial tissues with plenty of vessels or formation of blood lakes; and there was deposition of amyloid materials. Immunophenotypic analysis showed that all tumor cells were positive for CD79a and negative for CD20; that plasma cell marker (VS38C) was expressed in 41 cases; that light chain restriction was detected in all 43 cases( 31 for lamda and 12 for kappa), and that heavy chain expression was detected in 39 for IgG, 1 for IgA, 2 for both IgG and IgM, and 1 for IgA/IgG. CONCLUSION: The special histopathology in this group of solitary plasmacytomas is presented. The well-differentiated tumors should be distinguished from reactive plasmacytosis and MALT. And, when poorly differentiated solitary plasmacytomas were tentatively diagnosed, large cell lymphoma, melanoma and poorly differentiated carcinoma should be considered in differential diagnosis. PMID- 16468653 TI - [Effect of co-expressing M-bcr/abl and m-bcr/abl fusion gene transcripts on the clinical features of patients with chronic myelogenous leukemia at diagnosis]. AB - OBJECTIVE: To study the effect of co-expressing M-bcr/abl and m-bcr/abl fusion gene transcripts on the clinical features of the patients with chronic myelogenous leukemia (CML) at diagnosis. METHODS: m-bcr/abl fusion gene transcripts were detected by nested PCR. RESULTS: The percentage of the CML patients who co-expressed M-bcr/abl and m-bcr/abl was 51.4% at diagnosis. Comparison of the patients who co-expressed and those who expressed M-bcr/abl alone showed there was no difference in their hemoglobin concentrations, WBC counts, neutrophilic alkaline phosphatase (NAP) stains, Ph chromosomes and hepatosplenomegaly; however, significant difference in platelet count was seen. In the patients with b3a2 who co-expressed m-bcr/abl, an increased platelet count was noted, compared with that of patients with M-bcr/abl alone. But in patients with b2a2, the two groups showed no difference in all clinical features. Two patients had priapism, their m-bcr/abl being negative. CONCLUSIONS: CML patients who co-expressed b3a2 and m-bcr/abl showed a tendency to have an increased platelet count at diagnosis. Co-expressing m-bcr/abl has no the effect on the clinical features of CML patients with b2a2. PMID- 16468654 TI - [Distribution of fimA genotype of Porphyromonas gingivalis in Chinese periodontitis patients and its relationship with chronic periodontitis]. AB - OBJECTIVE: To investigate the distribution of fimA genotype of Porphyromonas gingivalis in Chinese periodontitis patients and try to find the relationship between fimA genotype and chronic periodontitis. METHODS: Subgingival plaque samples were collected from 101 periodontitis patients. P. gingivalis 16S rRNA primer and fimA type-specific primer were designed. The distribution of fimA genotype in periodontitis patients was detected by PCR. Clinical periodontal indices (PPD, CAL and BOP) were measured at the sample tooth's six points; namely, the mesio-, mid-, distobuccal and mesio-, mid-, distolingual points. RESULTS: P. gingivalis was detected in 89 periodontitis patients (88.1%). Among them, a single fimA genotype was detected in most subgingival plaque samples (65.1%), and the most prevalent fimA genotype was type II (43.8%), followed by type IV (40.4%); Type II fimA and IV fimA were more frequently detected in mild/moderate periodontitis group and severe periodontitis group. CONCLUSION: The findings indicate that P. gingivalis with type II fimA and IV fimA are more predominant in Chinese periodontitis, and the organisms are involved in the destructive progression of periodontitis in Chinese. PMID- 16468655 TI - [Expression of galectin-3 mRNA in gastric cancer with peritoneal metastasis]. AB - OBJECTIVE: To search for the role of galectin-3 gene in gastric cancer with peritoneal metastasis. METHODS: Using real-time quantitative RT-PCR for detecting the expression of galectin-3 mRNA in 35 matched samples, including primary cancer lesions, lymph node lesions, peritoneal metastasis and adjacent noncancerous mucosa. ANOVA and SNK-q test were used. RESULTS: The levels of mean expression of galectin-3 mRNA in gastric cancer (385.639 +/- 98. 534), in peritoneal metastasis (368.589 +/- 93.431) and in lymph node metastasis (375.920 +/- 94.346) were significantly higher than that in noncancerous mucosa (2.158 +/- 0.896); there were statistically significant differences respectively (P < 0.05). However, the levels of the expression in gastric cancer, in peritoneal metastasis and in lymph node metastasis were shown to be of no significant differences by multiple comparison, P > 0.05. The expression level in the group of gastric cancer was noted to have positive correlation with that in the group of peritoneal metastasis. These data indicated that the expression levels of galectin-3 in gastric cancer were well correlated to peritoneal metastasis. CONCLUSION: The overexpression of galectin-3 in gastric cancer is well correlated with peritoneal metastasis. Galectin-3 mRNA level may serve as a biological marker in the pre operative judgment on whether the gastric cancer has had peritoneal metastasis or has just the potential of metastasis. PMID- 16468656 TI - [Association between TGF-Beta1 in allograft and chronic allograft nephropathy]. AB - OBJECTIVE: To investigate the relation between TGF-beta1 in allograft and chronic allograft nephropathy (CAN). METHODS: The levels of urine TGF-beta1 were tested in 146 recipients whose renal function were normal from September 1, 2000 to January 31, 2001. Twenty recipients with the highest level of urine TGF-beta1 were classified in group A, while 20 other recipients with the lowest level of urine TGF-beta1 were classified in group B. In these two groups biopsies were carried out in 14 cases and 12 cases respectively, and TGF-beta1 mRNA in the biopsies was measured by RT-PCR. The levels of TGF-beta1 in the blood were also measured in the two groups. Three years later, the renal function was compared between the two groups. Biopsies were carried out in renal recipients whose creatinine is higher than normal. RESULTS: The level of TGF-beta1 in the blood showed no significant difference between the two groups; 3 years after transplantation, the loss of renal function in group A was severer than that in group B. The number levels of CAN cases in group A was larger than that in group B. The expression levels of TGF-beta1 and TGF-beta1 mRNA of the allografts were higher in group A than in group B; there were statistically significant differences between the two groups. CONCLUSION: The findings suggest that there is an association between TGF-beta1 in kidneys and CAN. The level of urine TGF beta1 after renal transplantation may predict future renal function. PMID- 16468657 TI - [Expression of Merlin in cortex of temporal lobe and in hippocampal CA1 region of the Kindling Model of Epilepsy induced by corciaria lactone in rats]. AB - OBJECTIVE: To explore the relationship between merlin and hippocampal sclerosis of temporal epilepsy. METHODS: The kindling model of epilepsy induced by corciaria lactone (CL) in rats was used. The expression of merlin in neuron of cortex of temporal lobe and hippocampal CA1 region was observed using immunohistochemistry method. Comparison of the amount of neuron with expression of merlin in the two locations was made between the kindled group, non-kindled group and control group. RESULTS: The expression of merlin in neuron of cortex of temporal lobe and hippocampal CA1 region of the kindled group was higher than the expression of the other two groups (P < 0.05), and there was no significant difference between the expression in the non-kindled group and that in the control group (P > 0.05). The expression of merlin in glial cell of the same region of all groups was seldom seen. CONCLUSION: The super-expression of merlin in neuron of cortex of temporal lobe and hippocampal CA1 region of the kindled rats may be involved in the process of neuronal apoptosis and hippocampal sclerosis. PMID- 16468659 TI - [Inflammatoin, oxidative stress and carbonyl stress in uremic patients]. AB - OBJECTIVE: To evaluate the states of inflammation, oxidative stress and carbonyl stress in uremic patients and analyze their relationships. METHODS: One hundred and twenty-eight cases were divided into 6 groups: non-dialysis uremia group (n=25), peritoneal dialysis group (n=19), hemophan(Hem) membrane dialysis group (n=25), polyamide (PS) membrane dialysis group (n=25), diabetes with normal renal function group (n= 23) and normal control group (n=11). Spectrophotometry and immune turbidimetry were used to measure the serum SOD, VitC, VitE, MDA and total carbonyl compounds (TCC) levels. RESULTS: Compared with non-uremia groups, the uremia groups had lower serum SOD, VitE and VitC levels, but higher CRP, MDA and TCC levels (P < 0.01). The peritoneal dialysis group had higher SOD, VitC, VitE levels in comparison with the Hem group, but did not differ from PS group in those levels (P > 0.05). The MDA and TCC levels in the peritoneal dialysis group were lower than those in the two hemodialysis groups, but there were no significant differences in CRP level between the groups. Compared with Hem group, the PS group had higher VitC, VitE levels, higher TCC clearance, and the same SOD, MDA, CRP and pre-dialysis TCC levels. When the variables were analyzed with TCC, the results of multi-variate regression showed that the standardized coefficients were MDA (0.727, P < 0.01), CRP (0.370, P < 0.01), SOD (0.192, P < 0.05), VitC (-0.153, P < 0.01), VitE (0.054, P = 0.30) respectively. CONCLUSION: Uremic patients are in inflammatory, oxidative-stress and carbonyl-stress states. Inflammation and oxidative stress are probably the important mechanism of carbonyl stress. It is not yet clear whether dialysis methods can influence uremic inflammatory, oxidative-stress or carbonyl-stress state. PMID- 16468658 TI - [Expression of matrix metalloproteinase and tissue inhibitor of metalloproteinase in adenomyosis]. AB - OBJECTIVE: To investigate the expressions of matrix metalloproteinase -2, -3 (MMP 2,-3) and tissue inhibitor of metalloproteinase-2 (TIMP-2) in internal uterine endometriosis tissue and endometrium from women with and without endometriosis throughout the menstrual cycle. METHODS: Immunohistochemical staining of tissues was performed to study the expressions and locations of MMP-2, MMP-3, TIMP-2 between women with and without endometriosis. The real-time PCR technique was applied to detected the mRNA expressions of MMP-2, MMP-3 and TIMP-2. RESULTS: We found that ectopic endometrium from patients with endometriosis expressed higher levels of MMP-2 and MMP-3 and lower levers of TIMP-2 than normal controls did (P < 0.05). MMP-2 and MMP-3 were detected strongly in both stromal and epithelial cells of ectopic endometrium from patients with endometriosis, when mostly detected in the epithelial cells in the control group. All eutopic and ectopic endometrium samples from women with and without endometriosis throughout the menstrual cycle showed similar expressions of MMP-2, MMP-3 and TIMP-2. Quantitative expressions of MMP-2 mRNA, MMP-3 mRNA and TIMP-2 mRNA were significantly lower in eutopic endometrium from controls compared with ectopic endometrium from patients with endometriosis (P < 0.05). Eutopic endometrium from controls in the proliferative phase showed significantly increased expressions of MMP-2 mRNA compared with that in the secretory phase (P < 0.05). CONCLUSION: The results suggest that ectopic endometrium with adenomyosis has more invasiveness and is prone to peritoneal implantation maybe to involved in the high expressions of MMP-2 and MMP-3 and the less expressions of TIMP-2 than endometrium from women without adenomyosis does. PMID- 16468660 TI - [Expression of deltaNp63 in salivary gland tumours]. AB - OBJECTIVE: To detect the expression of deltaNp63 in human salivary gland tumor and analyze its role in the malignant salivary gland tumors. METHODS: Collecting the samples from 68 cases of pathologically proven salivary gland tumours and investigating the microscopic sections with HE stain and immunohistochemical-SP stain. RESULTS: It was found that the expression of deltaNp63 is gradually increased in the malignant tumours, the myoepithelial cells and the basal cells of the salivary gland tumors are positive, and the expression of deltaNp63 is correlated directly with benign and malignant tumor. CONCLUSION: deltaNp63 plays an important role in salivary gland tumours. deltaNp63 possesses some special activity that is characteristic of oncogene. p63 is a sensitive and highly specific marker of myoepithelial cells in salivary gland tumours and an additional marker for defining myoepithelial histogenesis. p63 is of definite clinical value in falicitating diagnosis and differential diagnosis. PMID- 16468661 TI - [Analysis of the movement of long axis and the distribution of principal stress in abutment tooth retained by conical telescope]. AB - OBJECTIVE: To study the movement of long axis and the distribution of principal stress in the abutment teeth in removable partial denture which is retained by use of conical telescope. METHODS: An ideal three dimensional finite element model was constructed by using SCT image reconstruction technique, self programming and ANSYS software. The static loads were applied. The displacement of the long axis and the distribution of the principal stress in the abutment teeth was analyzed. RESULTS: There is no statistic difference of displacenat and stress distribution among different three-dimensional finite element models. Generally, the abutment teeth move along the long axis itself. Similar stress distribution was observed in each three-dimensional finite element model. The maximal principal compressive stress was observed at the distal cervix of the second premolar. CONCLUSION: The abutment teeth can be well protected by use of conical telescope. PMID- 16468662 TI - [Proteomic analysis of proteins related to retinoid acid resistance]. AB - OBJECTIVE: To evaluate the applicability of proteomic methods for shedding light on the mechanisms of All-trans retinoic acid resistance. METHODS: The expression of cellular proteins in the retinoid acid sensitive cell line NB4 and retinoid acid resistant cell line MR2 was analyzed using the two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Differentially expressed proteins were analyzed by mass spectrometry for peptide mass finger and identified by SWISS-PROT protein database. RESULTS: Approximately 600 spots appeared in an sodium dod(SDS-GEL). The match of the expression of cellular proteins between NB4 and MR2 was better. Three significantly differentially expressed proteins were screened and identified to DJ-1 and HSP70, as well as KIAA1289 of which the function is not yet known. CONCLUSION: The utilization of 2D-PAGE, coupled with mass spectrometry is effective in screening the drug resistance-associated proteins and could provide molecular targets for the elucidation of resistance mechanisms. PMID- 16468663 TI - [Separation and culture of mouse embryonic palatal mesenchymal cells in vitro]. AB - OBJECTIVE: To modify the operation of dissecting embryonic palatal shelves and purify the mouse embryonic palatal mesenchymal (EPM) cells in primary culture. METHODS: The embryonic palatal shelves were dissected using a surgical microscope by modified operation. Then the embryonic palatal shelves were incubated with Dispase and the isolated EPM cells were cultured. Immunofluorescence technique was used to identify the characteristics of cells. RESULTS: Embryonic palatal shelves could be dissected accurately and easily with a modified operation. The purified EPM cells contained scarcely epithelial cells. EPM cells were anti-HNK 1, S-100, vimentin positive and anti-CK negative. CONCLUSION: A modified method for dissecting embryonic palatal shelves and purifying the EPM cells of primary culture was established. PMID- 16468664 TI - [Long-term culture and differentiation of human glioma stem cells ]. AB - OBJECTIVE: To explore the culture and subculture conditions for glioma stem cells(GSCs) and to investigate the differentiation potential of GSCs. METHODS: The cells from human glioma were mechanically dissociated. Cells were cultured in N2 or B27 medium with basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF), and they were identified by immunocytochemistry. RESULTS: Glioma stem cells from human glioma have been successfully cultured. They formed typical neurospheres in suspension, and they could be cultured and passaged steadily in vitro. The majorities of the cells expressed vimentin and nestin, which were the markers for neural stem cells. They could differentiate into neurons and astrocytes, and express glial fibrillary acidic protein and beta III-tubulin respectively. CONCLUSION: Human glioma stem cells could be cultured from gliomas in vitro, and they could differentiate into neurons and astrocytes, thus providing a basis for further studies. PMID- 16468665 TI - [The pharmacokinetic study on cytarabine nanoparticle lyophilization injection in rabbits]. AB - OBJECTIVE: The pharmacokinetics of Cytarabine Polybutylcyanoacrylate Nanoparticles (Ara-C-PBCA-NP) lyophilization injection in rabbits was studied. METHODS: Ara-C water solution was taken as reference, the concentration of Ara-C was determined by HPLC, and the pharmacokinetic parameters were calculated. RESULTS: Pharmacokinetic properties of Ara-C water solution and Ara-C-PBCA-NP lyophilization injection complied with two-department model. The parameters t1/2 beta and MRT of Ara-C-PBCA-NP lyophilization injection were prolonged, and CL was reduced dramatically (P < 0.05). CONCLUSION: Ara-C-PBCA-NP lyophilization injection possesses prolonged retention time in vivo and significant sustained releasing character. PMID- 16468666 TI - I'm sick of being me: developmental themes in a suicidal adolescent. AB - Adolescence is a developmental transition period during which there are profound transformations in emotional, cognitive, and behavioral systems. Despite being a time of rapid development and increasing rates of suicidality, limited research has examined possible interrelationships. Through the use of a case study, this paper illustrates the role of developmental factors in a teenage girl's experience becoming and overcoming being suicidal. The processes of cognitive development, identity formation, and autonomy-seeking are discussed within the context of her story. PMID- 16468667 TI - A framework for studying minority youths' transitions to fatherhood: the case of Puerto Rican adolescents. AB - A theoretical framework is proposed for studying minority young men's involvement with their babies that combines the integrative model of minority youth development and a life course developmental perspective with Lamb's revised four factor model of father involvement. This framework posits a relationship between demographic and family background variables (such as education, employment, income, and family of origin) and fatherhood outcomes moderated by personal characteristics (such as sex-role ideology, acculturation, risk taking, and alienation) and mediated by definitions of fatherhood, life priorities normative for the culture under study, and sexual behavior. Once there is an acknowledged infant, a father's involvement is influenced by child characteristics, perceived fathering competence, social support, and quality of the relationship with the mother. PMID- 16468668 TI - Male and female delinquency trajectories from pre through middle adolescence and their continuation in late adolescence. AB - This study of male and female adolescent delinquency trajectories focuses on the prediction of late adolescence delinquency, based on earlier delinquency and social support. In this 3-wave longitudinal survey, 270 Dutch adolescents (113 males and 157 females) ages 12 to 14, were followed for a period of 6 years. For males, the level of delinquent activity in late adolescence strongly depends on earlier delinquent activities (R2 = .33, p < .0005). In contrast, the level of female delinquency in late adolescence is far less predictable (R2 = .18, p < .001), and could not be predicted from delinquent activities during pre and early adolescence, while support from the mother during late adolescence was associated with reduced delinquency for females. Different models may be needed to explain the development of delinquency for males versus females. PMID- 16468669 TI - Optimistic bias among potential perpetrators and victims of youth violence. AB - This study furthers the current understanding of optimistic bias regarding youth violence among high school students. Results from a survey of 387 urban high school students indicate a wide range of predictors of optimistic bias, including experience, demographics, and attitudes. Linkages to other developmental frameworks (personal fable and self-efficacy) suggest future directions for additional research. PMID- 16468670 TI - A review of research on the effects of religion on adolescent tobacco use published between 1990 and 2003. AB - An electronic search of Medline and PsycInfo produced 29 studies that specifically investigated the effects of religion on adolescent tobacco use. Independent (religion) and dependent (tobacco use) variables and variables controlled for in statistical analyses were categorized. Twenty-two of the 29 studies reported at least one significant effect of religion on tobacco use, with 31 of 43 separate analyses of religious variables yielding significant negative correlations between religion and tobacco use. Religion was inversely related to all measures of tobacco use (lifetime, occasional, and regular use), but the findings suggest religion's primary effect is its prohibitive influence against ever using tobacco. PMID- 16468671 TI - Family functioning and early onset of sexual intercourse in Latino adolescents. AB - The purpose of this study was to identify factors associated with early onset of sexual intercourse. Within an ecological system's conceptual framework, familial factors associated with early onset of sexual activity were identified in a sample of 425 adolescents from San Juan metro area schools. Measures included questions about sexual activity, sexual permissiveness, and such familial variables as: discipline, parental supervision, and parental support. Significant relationships were observed between early onset of sexual intercourse and parental supervision, discipline, parental support, and parents' marital status. Results suggest the key role of parents and family in prevention of HIV-risk behaviors among adolescents in terms of delaying sexual onset. Overall, the study described youths who postponed sexual activity as having greater support, supervision, and parental involvement. PMID- 16468672 TI - An existential view of adolescent development. AB - In clinical work with adolescents there is a stark similarity between what they experience and the concepts of existentialism. However, surprisingly very little has been written in terms of how the concepts of existentialism can or should be applied to this age group. Rather, existentialism seems to be a concept reserved for its application to the adult population. In fact, a search of the literature included in PsychINFO under "existentialism" and "adolescent development" results in only 5 hits. Perhaps by avoiding the use of these ideas as they might apply to adolescents, for various reasons a disservice is being done. Drawing on ideas posited in existential literature will not only benefit adolescents, it will assist caring adults in their work with them and encourage professionals to expand on existing ideas when conducting research and developing theories that apply to this age group. PMID- 16468673 TI - An exploration of the attitudinal and perceptual dimensions of body image among male and female adolescents from six Latin American cities. AB - Using survey methodology, this exploratory study examined the attitudinal and perceptual dimensions of body image among 1,272 eighth- and ninth-grade males and females from higher and lower socioeconomic backgrounds in Buenos Aires, Argentina (n = 195), Guatemala City, Guatemala (n = 212), Havana, Cuba (n = 213), Lima, Peru (n = 218), Panama City, Panama (n = 195), and Santiago, Chile (n = 239). The two dimensions of body image were assessed by showing the participants nine male and nine female silhouettes depicting body sizes ranging from extremely thin (#1) to extremely obese (#9). Approximately 50% of the male and female adolescents chose silhouette #3 as best depicting the attractive male body. Silhouette #2 was chosen by 45% of the females as best depicting feminine attractiveness, while 45% of the males chose silhouette #3. When identifying the silhouette that best depicted the healthy male body, 26% of the participants chose silhouette #2, 41% chose #3, and 19% chose #4. Silhouette #2 was selected by 33% of the adolescents as best depicting the healthy female body, 38% chose #3, and 14% chose #4. In all cities, the effect of body mass index (BMI) on body satisfaction was highly significant (p < 0.001). Almost 40% of the middle weight adolescents indicated a preference to be thinner, even though 72% of the males and 84% of the females in this subgroup already perceived themselves as thin. Similarly, 89% of the heavier weight participants indicated a preference to be thinner, even though only 10% of the males and 2% of the females in this subgroup perceived themselves as heavy. In five of the six cities, a significantly higher (p < 0.05) percentage of females than males desired to be thinner, the exception being Havana, where the distribution of desired change in body size was almost identical for males and females. Findings suggest a need for culturally sensitive, age- and gender-specific preventive interventions focusing on the importance of achieving a healthy weight during adolescence to promote body size acceptance among adolescents who are at a healthy weight. PMID- 16468674 TI - Psychometric properties of the Chinese Family Assessment Instrument in Chinese adolescents in Hong Kong. AB - This paper reports evidence on the factor structure, reliability, and validity of the Chinese Family Assessment Instrument (C-FAI), an instrument developed to assess family functioning in Chinese populations. A convenience sample of 1,462 adolescents from junior secondary schools completed the C-FAI and measures of parent-adolescent conflict. Exploratory and confirmatory factor analyses showed that five factors were abstracted from the scale, a finding which was similar to those reported in previous validation studies. Results also showed that the C-FAI was internally consistent and its scores were moderately correlated with measures of conflict with parents, providing support for the convergent validity of the measure. Finally, females perceived family functioning to be better than did males and age was linearly related to perceived family dysfunction. In conjunction with the previous findings, the present study suggests that the C-FAI can be used to objectively assess family functioning in Chinese adolescents. PMID- 16468675 TI - Mental health condition of the only-child: a study of urban and rural high school students in China. AB - The mental health of the only-child continues to generate interest in research literature. The present study examines the issue in China, where the one-child phenomenon is highest due to deliberate government policy. Subjects are 299 and 333 students in two high-rank high schools in urban Harebin and rural Qing an Xian, respectively (mean age = 17.2 years). Both locations are in the Heilongjiang Sheng Province of China. Results showed that urban only-children experienced significantly lower love awareness from family, higher neurotic and social depression, trait anxiety, perceived stressors, and interpersonal dependency than did urban non-only children. No significant differences were found in the rural only- and non-only children. Low love awareness from parents and peers was associated with high negative mental health conditions in the children. Low love awareness also precipitated perceived stressors which resulted in negative mental health in the covariance structure analysis. PMID- 16468676 TI - Parental attitudes and ego identity status of Turkish adolescents. AB - This study investigated the differences among 403 middle adolescents regarding Marcia's four identity statuses in terms of perceived parental attitudes and gender. The Extended Objective Measure of Ego Identity Status (EOMEIS-2) and Parental Education Scale (PAS) were used to collect data. Results showed that children of authoritative parents scored significantly higher on identity foreclosure than those of neglectful parents. Children of permissive parents scored significantly higher on identity foreclosure than those of neglectful parents. In addition, female students scored higher on achievement identity status while male students scored higher on identity foreclosure. PMID- 16468678 TI - Panning for gold: a clinician's guide to using research. AB - The gap between research and clinical practice is one of the key challenges facing family therapy. Clinicians often fail to incorporate research findings into their practice because they do not know how to search, evaluate, or apply research to their clinical work. The purpose of this article is to help clinicians become better consumers of research. This article explores the potential value of research to clinicians, as well as negative beliefs that clinicians may have about research. The article also describes how clinicians can use research to inform their clinical work, as well as potential challenges that can be encountered. PMID- 16468677 TI - Textual representations of diversity in COAMFTE accredited doctoral programs. AB - The use of the Internet is growing at a staggering pace. One significant use of the Internet is for potential students and the parents of potential students to explore educational possibilities. Along these lines potential marriage and family therapy students may have many questions that include a program's commitment to cultural diversity. This study utilized qualitative content analysis methodology in combination with critical race theory to examine how Commission On Accreditation for Marriage and Family Therapy Education (COAMFTE) accredited doctoral programs represented cultural text on their World Wide Web pages. Findings indicate that many COAMFTE-accredited doctoral programs re present programmatic information about diversity that appear to be incongruent with cultural sensitivity. These apparent incongruities are highlighted by the codification, inconsistent, and isolated use of cultural text. In addition, cultural text related to social justice was absent. Implications and suggestions are discussed. PMID- 16468679 TI - Supervising family therapy trainees in primary care medical settings: context matters. AB - The purpose of this article is to identify and describe four essential skills for effective supervision of family therapy trainees in primary care medical settings. The supervision skills described include: (1) Understand medical culture; (2) Locate the trainee in the treatment system; (3) Investigate the biological/health issues; and (4) Be attentive to the self-of-the-therapist. Recommendations are also made to help supervisors become better prepared for the questions medical family therapy trainees bring to supervision. PMID- 16468680 TI - Marriage and family therapist interns' experiences of growth. AB - Few studies have investigated therapist interns' growth experiences, both personal and professional, and the interplay between them. This qualitative study explored the reciprocal process of growth during clinical training: the personal life experiences that affect professional growth and the professional/clinical experiences that affect personal growth. Thirteen marriage and family therapist interns, using Internet discussion formats, identified numerous personal, clinical, and professional growth experiences. Implications of this study, illustrating the recursive and interactive flow between therapists' personal and professional growth, are presented. PMID- 16468681 TI - Practicing as a postmodern supervisor. AB - In this article, aspects of postmodern supervision are explored that highlight the blurring of boundaries that occurs between the multiple roles performed by supervisors. An approach to supervision is detailed that shows how a supervisor who navigates between roles can assist supervisees in constructing identities congruent with the stories they tell about themselves as successful therapists. Six of the many possible roles that supervisors play are illustrated through dialogue taken from a group supervision session. In the last part of this article, concepts that inform a postmodern approach to supervision, borrowed from the literature on narrative and constructionist therapy, are discussed. PMID- 16468682 TI - Supervisor multicultural competence and its relation to supervisory process and outcome. AB - This study investigated the direct and indirect effects of marriage and family therapy trainees' perceptions of their supervisors' multicultural competence in supervision on the supervisory working alliance, trainees' multicultural competence (case conceptualization abilities in etiology and treatment), and perceived supervision satisfaction. Path analyses revealed supervisor multicultural competence to be positively associated with supervisory working alliance and perceived supervision satisfaction. In addition, supervisor multicultural competence seemed to have a negative effect on trainee etiology conceptualization abilities. Finally, results suggested supervisory working alliance to be a significant mediator in the relationship between supervisor multicultural competence and supervision satisfaction. Findings are discussed within the context of theoretical, empirical, and practical implications for multicultural supervision. PMID- 16468683 TI - Burnout among mental health professionals: special considerations for the marriage and family therapist. AB - Burnout is a syndrome consisting of physical and emotional exhaustion resulting from negative self-concept, negative job attitudes, and loss of concern for clients. This research study explores potential predictors and prevalence of burnout among marriage and family therapists (MFTs). It evaluates the Maslach Burnout Inventory (MBI) to establish its applicability to MFTs. Our sample of 116 Clinical Members of the American Association for Marriage and Family Therapy responded to a mailed questionnaire including demographic information and the MBI. Overall, our sample reported low-to-moderate ranges of burnout. Differences were noted in degrees of burnout across job settings. Predictors of clinician burnout include hours worked per week and job setting. Factor analysis indicates that the MBI is an appropriate assessment tool for measuring burnout among MFTs. Implications for clinical practice are discussed. PMID- 16468684 TI - The relationship between marriage and family therapists and complementary and alternative medicine approaches: a national survey. AB - Respondents to a mail survey of a random sample (N = 424) of Clinical Members of the American Association for Marriage and Family Therapy provided information about their contexts of practice, use of complementary and alternative medicine (CAM), and relationships with CAM providers. Consistent with both national trends and the experience of psychologists as reported in a similar survey, the results of this survey suggest that marriage and family therapists have been affected significantly by and have a growing awareness of CAM practices. Limitations of the study and implications for the field are discussed. PMID- 16468686 TI - Strategies for architectural integration of ventilated caging systems. PMID- 16468685 TI - The relationship between marriage and family therapists and complementary and alternative medicine approaches: a qualitative study. AB - In this article, we delineate the qualitative phase of a mixed-method research study focused on understanding the relationship between Clinical Members of the American Association for Marriage and Family Therapy (MFT) and complementary alternative medicine (CAM). Based on an analysis of the data derived from telephone interviews with 54 respondents, we describe four themes: definitional issues, depth of awareness of CAM, fit with MFT, and ethical considerations. Our discussion focuses on the findings of this phase, considerations from the quantitative phase, and reflections on the research study as a whole. While acknowledging the limitations of the study, we conclude that the growing awareness of and involvement with CAM approaches and practitioners among MFTs suggest a need for further education for both professionals and clients. We also note the importance of additional research support for the use of CAM practices. PMID- 16468687 TI - Microenvironments in Microisolation Cages Using BALB/c and CD-1 Mice. PMID- 16468689 TI - Comparison of Two Enzyme-linked Immunosorbent Assays for Detecting Antibody to Rodent Viruses. PMID- 16468688 TI - Effect of the New USDA Cage Height Standard on Body Weights of Weanling Djungarian Dwarf Hamsters (Phodopus sungorus). PMID- 16468690 TI - Analysis of Commercial Laboratory Rat Rations for Fumonisin B1, a Mycotoxin Produced on Corn by Fusarium moniliforme. PMID- 16468692 TI - Performance coaching. PMID- 16468691 TI - Use of natural and provided shade by free-ranging rhesus monkeys on raccoon key, Florida: management considerations. PMID- 16468693 TI - Institutional Animal are and Use Committee Review of Animal Care and Use Programs. PMID- 16468694 TI - Evaluation of continuing education in the health professions. PMID- 16468695 TI - A method for jugular blood collection in rabbits. PMID- 16468696 TI - A Noninvasive, Cage-side Electrocardiogram and Blood Pressure Toxicology Screening Method for Beagles. PMID- 16468697 TI - Mite infestation in a rat colony. PMID- 16468698 TI - Elimination of Giardia muris from Syrian Hamsters. PMID- 16468699 TI - Reduction and refinement in animal usage for acute drug disposition study in Swine by use of multi-source sequential sampling. PMID- 16468700 TI - Rearing of crickets in quantity for feeding lab animals. PMID- 16468701 TI - Effectiveness of hand cleaning in sanitizing rabbit cages. PMID- 16468702 TI - Chronic jugular vein cannulation in beagles. PMID- 16468703 TI - Estimating maintenance feeding requirements for individually housed rhesus monkeys. PMID- 16468704 TI - Overview of hand washing by personnel in animal laboratory facilities. PMID- 16468705 TI - Validating a newly constructed animal facility. PMID- 16468706 TI - A Per diem Rate Comparison, Interrelating Budget Size, Percent Recovery, Facility Size, Accreditation, and Cost Accounting Factors. PMID- 16468707 TI - Characterization of preexisting humoral immunity specific for two cancer-testis antigens overexpressed at the mRNA level in non-small cell lung cancer. AB - In order to establish a rationale for immunotherapy for lung cancer, we have investigated immunological characteristics of tumor-associated antigens (TAAs) discovered through molecular approaches. Preexisting Abs specific to these predicted TAAs were examined using specimens of lung pleural effusions (LPEs) and sera in non-small cell lung cancer (NSCLC) patients. The novel cancer-testis (CT) antigens L514S and L552S were highly expressed in approximately half of the NSCLC tissues and established cell lines examined. When lung cancer patients in the USA and Japan were screened, 13%, 17%, and 5% were found to have Abs specific to recombinant L514S, L552S, and NY-ESO-1 proteins, respectively, whereas 48 normal donors had no Abs to these three CT antigens. The Ab titers specific to recombinant L552S and L514S proteins were similar to, and slightly lower than, Abs specific to NY-ESO-1 in stage IV NSCLC patients. To further characterize the preexisting specific Abs, the epitopes were analyzed using 20-aa length peptides entirely covering both antigens. An epitope common to the patients' L514S specific Abs was identified as aa 85-100 and multiple epitopes, including a major epitope (aa 141-160), were identified for L552S-specific Abs. The Ab epitopes thus identified are not found in human, animal, or bacterial proteins, other than L514S, L552S, or XAGE-1. These data clearly demonstrate that both molecularly defined CT antigens L514S and L552S are immunogenic, at least in terms of humoral responses, suggesting that both CT antigens are promising candidates for immunotherapy. PMID- 16468708 TI - Calix[4]arene alpha-aminophosphonic acids: asymmetric synthesis and enantioselective inhibition of an alkaline phosphatase. AB - [structure: see text] Chiral calix[4]arene alpha-aminophosphonic acids were obtained through diastereoselective Pudovik-type addition of sodium ethyl phosphites to the chiral calixarene imines, removal of chiral auxiliary groups, and mild dealkylation of phosphonate fragments. The diacids obtained show inhibitory activity toward porcine kidney alkaline phosphatase that depends considerably on the absolute configuration of the alpha-carbon atoms. PMID- 16468709 TI - Selective one-pot synthesis of trithiocarbonates, xanthates, and dithiocarbamates for use in RAFT/MADIX living radical polymerizations. AB - [reaction: see text] We report a facile route for the production of chain transfer agents for reversible addition fragmentation chain transfer (RAFT) and macromolecular design via the interchange of xanthates (MADIX) polymerizations, via a one-pot reaction. 1,1'-Thiocarbonyl diimidazole (TCDI) undergoes controlled monosubstitution when reacted with secondary thiols or alcohols. The intermediate S/O-esters of imidazole-N-thionocarboxylic acid react efficiently with a range of primary thiols, alcohols, and amines to form asymmetrical dithiocarbonates, trithiocarbonates, and dithiocarbamates, respectively. The synthesis provides a facile approach to the controlled radical polymerization of vinyl monomers through the reversible addition-fragmentation chain transfer (RAFT) mechanism. PMID- 16468710 TI - Very efficient, reusable copper catalyst for carbene transfer reactions under biphasic conditions using ionic liquids. AB - [reaction: see text] The complex {[HC(3,5-Me(2)pz)(3)]Cu(NCMe)}BF(4) catalyzes the transfer of the :CHCO(2)Et unit from ethyl diazoacetate to several saturated and unsaturated substrates with very high yields and under biphasic conditions using the ionic liquid [bmim][PF(6)] and hexane as the reaction medium. The catalyst has been tested for several cycles of recovery and reuse without any loss of activity. PMID- 16468711 TI - A new radical-based route to calothrixin B. AB - [structure: see text] A high-yielding totally regioselective intramolecular homolytic acylation of a quinoline ring constitutes the key step in a new synthesis of the pentacyclic indolo[3,2-j]phenanthridine alkaloid calothrixin B. PMID- 16468712 TI - Development of a novel, highly efficient halide-catalyzed sulfenylation of indoles. AB - [reaction: see text] The reaction of a variety of indoles with N-thioalkyl- and N thioarylphthalimides to produce 3-thioindoles is reported. Catalytic quantities of halide-containing salts are crucial to the success of this reaction. This highly efficient reaction provides sulfenylated indoles from bench-stable, readily available starting materials in good to excellent yields. PMID- 16468714 TI - Nickel-catalyzed rearrangement of 1-acyl-2-vinylcyclopropanes. A mild synthesis of substituted dihydrofurans. AB - [reaction: see text] Mild Ni(0)-catalyzed rearrangements of 1-acyl-2 vinylcyclopropanes have been developed. The room-temperature isomerizations afford dihydrofuran products in yields regularly greater than 90%. A highly substituted, stereochemically defined cyclopropane was employed in the rearrangement to evaluate the reaction mechanism. Product analysis indicates that the overall reaction proceeds with retention of configuration at the vinyl bearing stereogenic center. PMID- 16468713 TI - Synthesis and properties of a novel phenylazomethine dendrimer with a tetraphenylmethane core. AB - [structure: see text] A new type of phenylazomethine dendrimer with a tetraphenylmethane core was synthesized by a convergent method. The properties of the dendrimer were confirmed by thermal, rheological, TEM, and AFM measurements. A stepwise radial complexation was clearly observed with SnCl(2). PMID- 16468715 TI - Phenacenes from Diels-Alder trapping of photogenerated o-xylylenols: phenanthrenes and benzo[e]pyrene bisimide. AB - [structure: see text] The synthesis of phenanthrene and benzo[e]pyrene bisimides, 1 and 2, was accomplished via the Diels-Alder trapping of sterically congested o xylylenols photochemically generated from 3,6-dibenzoyl-o-xylene and 1,4 dibenzoyl-9,10-dihydroanthracene, respectively. Absorption and emission from 2 are red-shifted from 1 and unsubstituted benzo[e]pyrene. The fluorescence quantum yield for 2 is an order of magnitude lower than that of 1 and comparable to that of the parent benzo[e]pyrene. PMID- 16468716 TI - The Pennsylvania Green Fluorophore: a hybrid of Oregon Green and Tokyo Green for the construction of hydrophobic and pH-insensitive molecular probes. AB - [structure: see text] Fluorescent small molecules are powerful tools for exploring cellular biology. As a more hydrophobic, photostable, and less pH sensitive alternative to fluorescein, we synthesized Pennsylvania Green, a bright, monoanionic fluorophore related to Oregon Green and Tokyo Green. Comparison of membrane probes comprising N-alkyl-3beta-cholesterylamine linked to 4-carboxy-Tokyo Green (pK(a) approximately 6.2) and 4-carboxy-Pennsylvania Green (pK(a) approximately 4.8) revealed that only Pennsylvania Green was highly fluorescent in acidic early and recycling endosomes within living mammalian cells. PMID- 16468717 TI - Self-assembling of n-type semiconductor tri(phenanthrolino)hexaazatriphenylenes with a large aromatic core. AB - [reaction: see text] Large disk-shaped aromatic tri(phenanthrolino)hexaazatriphenylenes 5a, 5b, and 5c with six butyl, dodecyl, and 4-octylphenyl groups, respectively, were self-assembled both in solution and film state to form one-dimensional aggregates. Their n-type semiconducting nature was indicated from CV measurement, in which the first reduction potentials were evaluated at around -1.7 V (vs Fc/Fc(+)) in dichloromethane. PMID- 16468719 TI - Synthesis of alpha-onoceradiene-like terpene dimers by intermolecular metathesis processes. AB - [reaction: see text] New alpha-onoceradiene analogues having a terpene homodimer skeleton are accessible from Weinreb's amide 2 derived from commercial (R)-(+) sclareolide using an intermolecular metathesis reaction as the key step to build the linker joining both terpene moieties. PMID- 16468718 TI - Correlation of F0F1-ATPase inhibition and antiproliferative activity of apoptolidin analogues. AB - [structure: see text] Apoptolidin (1) exhibits potent and highly selective apoptosis inducing activity against sensitive cancer cell lines and is hypothesized to act by inhibition of mitochondrial F(0)F(1)-ATP synthase. A series of apoptolidin derivatives, including a new intermolecular Diels-Alder adduct, were analyzed for antiproliferative activity in E1A-transformed rat fibroblasts. Potent F(0)F(1)-ATPase inhibition was not a sufficient determinant of antiproliferative activity for several analogues, suggesting the existence of a secondary biological target or more complex mode of action for apoptolidin. PMID- 16468720 TI - Formal enantioselective synthesis of (+)-compactin. AB - [reaction: see text] The challenging structural features and important biological activity of (+)-compactin (1) explain the substantial synthetic interest that it has generated. We report a novel enantioselective approach to the advanced intermediate 2a, which constitutes a formal synthesis of (+)-1. The sequence utilizes MacMillan's organocatalytic Mukaiyama-Michael reaction, which stereoselectively adds the silyloxyfuran 6 to alpha,beta-unsaturated aldehyde 7. The chirality generated in this reaction guides the formation of the other three consecutive stereocenters found in 2a. PMID- 16468721 TI - Synthesis of the erythrina alkaloid 3-demethoxyerythratidinone. Novel acid induced rearrangements of its precursors. AB - [reaction: see text] A new strategy for the synthesis of 3 demethoxyerythratidinone has been developed and is based on an extraordinarily facile intramolecular Diels-Alder reaction of a 2-imido-substituted furan. During the course of the synthesis, several novel acid-induced rearrangement reactions were encountered. PMID- 16468722 TI - Novel linear and tribranched polymers with redox-active end groups via W(CO)6 initiated metathesis polymerization. AB - [reaction: see text] Synthesis, characterization, and electrochemistry of linear and tribranched polyphenylacetylenes having redox-active ferrocene and/or (arene)chromiumtricarbonyl as end groups are reported in this study. The methodology adopts polymerization of phenylacetylene by a metathesis pathway, initiated by W(CO)(6) when photolyzed. A distance-dependent electronic communication between the metal centers is demonstrated in these polymers by means of cyclic voltammetry. PMID- 16468723 TI - Simple protocol for NMR analysis of the enantiomeric purity of primary amines. AB - [reaction: see text] A practically simple three-component chiral derivatizing protocol for determining the enantiopurity of 13 chiral primary amines by (1)H NMR spectroscopic analysis is described, including analysis of those that contain remote stereocenters. PMID- 16468725 TI - Magnesiated unsaturated silylated cyanohydrins as synthetic equivalents of aromatic and heterocyclic grignard reagents bearing a ketone or an aldehyde. AB - [reaction: see text] The preparation of iodo-substituted aryl, heteroaryl, or cycloalkenyl ketones as silylated cyanohydrins allows the smooth performance of an I/Mg-exchange using i-PrMgCl.LiCl. A facile deprotection of the resulting functionalized products obtained by a reaction with electrophiles (acid chlorides, allylic bromide, benzylidene-p-toluenesulfonamide, and 3 iodocyclohexenone) produces polyfunctional ketones in good overall yields. This sequence can be extended to aromatic iodoaldehydes. In these cases, the deprotection of the silylated cyanohydrin functionality is best performed with aqueous CuSO(4) under basic conditions. PMID- 16468724 TI - Unequivocal synthesis of (Z)-alkene and (E)-fluoroalkene dipeptide isosteres to probe structural requirements of the peptide transporter PEPT1. AB - [reaction: see text] Described is a novel synthetic route for dipeptide isosteres containing (Z)-alkene and (E)-fluoroalkene units as cis-amide bond equivalents via organocopper-mediated reduction of gamma-acetoxy- or gamma,gamma-difluoro alpha,beta-unsaturated-delta-lactams. The synthesized isosteres were evaluated in terms of their affinities for the peptide transporter PEPT1. trans-Amide isosteres tended to possess higher affinities for PEPT1 as compared to the corresponding cis-amide bond equivalents. PMID- 16468726 TI - Palladium-catalyzed multistep reactions involving ring closure of 2 iodophenoxyallenes and ring opening of bicyclic alkenes. AB - [reaction: see text] An efficient ring closure of 2-iodophenoxy-, 2-iodobenzyloxy , and 2-iodobenzylaminoallenes followed by ring opening of oxabenzonorbornadienes leading to the synthesis of 2-benzofuranyl, 1H-isochromenyl, or 1,2 dihydroisoquinoline methyl-1,2-dihydro-1-naphthalenol derivatives catalyzed by palladium complexes is described. PMID- 16468727 TI - FeCl3-Catalyzed aminohalogenation of arylmethylenecyclopropanes and arylvinylidenecyclopropanes and corresponding mechanistic studies. AB - [reaction: see text] The aminochlorination of methylenecyclopropanes (MCPs) 1 and vinylidenecyclopropanes (VCPs) has been explored with use of FeCl(3) (20 mol %) as a Lewis acid catalyst in acetonitrile under convenient mild conditions. The stereochemistry has been unambiguously confirmed by X-ray structural analysis. The aziridinium-based mechanism, accounting for both regio- and stereoselectivity, has been carefully studied. A linear free-energy relationship study of this reaction confirms consistency with the Hammet equation. PMID- 16468728 TI - Modified (22pi) smaragdyrins with large two-photon absorption cross section: a structure function correlation. AB - [structure: see text] Two-photon absorption (TPA) cross-section values for a series of 22pi smaragdyrins bearing phenylacetylenylphenyl and [(phenylacetylenyl)phenylacetylenyl]phenyl meso links and their Rh(I) derivatives are reported. PMID- 16468730 TI - Stereoselective synthesis of (E)-alpha,beta-unsaturated esters via carbene catalyzed redox esterification. AB - [reaction: see text] Stereoselective, carbene-mediated redox esterification of alkynyl aldehydes provides mild and atom economical access to (E)-configurated, alpha,beta-unsaturated carboxylic esters. The organocatalytic method relies on the generation of activated carboxylates via extended/conjugated umpolung in the presence of catalytic amounts of carbene precursor and base. PMID- 16468729 TI - Allylation of aldehydes and imines: promoted by reuseable polymer-supported sulfonamide of N-glycine. AB - [reaction: see text] A allylation of aldehydes and imines (generated in situ from aldehydes and amines) with allyltributyltin promoted by recoverable and reusable the polymer-supported sulfonamide of N-glycine has been developed. Good to high yields were obtained in various cases. Most of the SnBu(3) residue can be recovered as Bu(3)SnCl. Highly stereoselective synthesis of N-Boc-(2S,3S)-3 hydroxy-2-phenylpiperidine 7 was achieved by using the P4a-mediated allylation of Boc-l-phenylglycinal as a key step. PMID- 16468731 TI - Rates of C-S bond cleavage in tert-alkyl phenyl sulfide radical cations. AB - [reaction: see text] Radical cations of tert-alkyl phenyl sulfides 1-4 have been generated photochemically in MeCN in the presence of the N methoxyphenanthridinium cation (MeOP(+)), and the rates of C-S bond cleavage have been determined by laser flash photolysis. PMID- 16468732 TI - Electronic effects of ring substituents on triplet benzylic biradicals. AB - [reaction: see text] UV irradiation of alpha-(o-alkylphenyl)acetophenones with a methoxy or cyano substituent para to the o-alkyl group of the alpha-aryl ring has revealed that a methoxy group slightly increases the stereoselectivity but not the quantum yield of indanol formation, whereas a cyano group greatly lowers both diastereoselectivity and quantum efficiency, confirming the likelihood that hydrogen-bonding of the hydroxy group to the alpha-phenyl ring plays an important role in the cyclization of the photogenerated triplet 1,5-biradical intermediates. PMID- 16468733 TI - Synthesis of 1-deoxysphingosine derivatives with conformationally restricted pyrrolidinediol head groups. AB - [structure: see text] A family of cyclic 1-deoxysphingolipid derivatives of structure 4 has been designed and synthesized, which may serve as tumorigenesis suppressors for various cancers. Compound 4 is a second-generation analogue developed from sphingosine (1), in which a hydroxyl substituent is moved from C1 to C5 and a methylene is added for conformational rigidity between the C2 nitrogen substituent and C4. The synthetic chemistry for pyrrolidine ring closure at C3-C4 features ring-closing metathesis followed by hydroboration-oxidation. PMID- 16468734 TI - CuI-Catalyzed tandem intramolecular amidation using gem-dibromovinyl systems. AB - [reaction: see text] Imidazoindolones are present as the key structural motif in the family of antifungals, fumiquinazolines, and the antagonist asperlicin. The first example of a CuI-catalyzed tandem intramolecular amidation forming substituted imidazoindolones from readily accessible ortho gem dibromovinylanilines is described. PMID- 16468735 TI - Novel and efficient synthetic path to proaporphine alkaloids: total synthesis of (+/-)-stepharine and (+/-)-pronuciferine. AB - [reaction: see text] A novel synthetic path to proaporphine alkaloids was established by employing aromatic oxidation with a hypervalent iodine reagent, where an unprecedented carbon-carbon bond forming reaction between the para position of a phenol group and an enamide-carbon took place smoothly to give the desired spiro-cyclohexadienone. PMID- 16468736 TI - Total synthesis, structural revision, and absolute configuration of (-) clavosolide [corrected] A. AB - [reaction: see text] Enantioselective synthesis of 3, a revised structure for clavosolide A, was completed. Both (1)H and (13)C NMR spectra of the natural and synthetic compounds were identical, and optical rotation measurements identified the absolute configuration of the natural clavosolide A as [corrected] 3. PMID- 16468738 TI - Regio- and diastereoselective reductive coupling of vinylepoxides catalyzed by titanocene chloride. AB - [reaction: see text] The Ti(III)-catalyzed reaction of a series of vinylepoxides leads, with regio- and E-diastereoselectivity control, to good-to-excellent yields of the corresponding homocoupling products. This homocoupling reaction, which involves a new C-C bond-forming method, takes place via a S(N)2' process between an allyltitanium species and the starting vinylepoxide. The process can be used for the rapid and efficient formation of highly valuable intermediates for organic synthesis, as well as new interesting homologues of natural products. PMID- 16468737 TI - Catalytic asymmetric prins cyclizations: cation generation and trapping with (BINAP)Pt dications. AB - [reaction: see text] (R)-[(tolBINAP)Pt(NC(6)F(5))(2)][SbF(6)](2) (5) catalyzes the highly enantioselective Prins reaction between 2-allylphenols and glyoxylate esters. Other Lewis acid catalysts favor glyoxylate-ene products. PMID- 16468739 TI - General method for synthesizing pyranoid glycals. A new route to allal and gulal derivatives. AB - [reaction: see text] Pyranoid glycals of all configurations can be obtained from pentoses through an olefination-cyclization-elimination sequence. The elimination can be carried out with excellent yields under radical conditions or by using common reductive reagents such as Zn/Cu, TiCl(4)/LiAlH(4), or lithium naphthalenide. The proposed method is appropriate for the synthesis of glycals with allo or gulo configurations because the cyclization step is more efficient for these substrates. PMID- 16468740 TI - Enantioselective total synthesis of convolutamydines B and E. AB - [reaction: see text] The first enantioselective total synthesis of convolutamydines B and E has been achieved using our vinylogous Mukaiyama aldol reaction. The synthesis features highly diastereoselective vinylogous Mukaiyama aldol reaction with isatin instead of aldehydes to construct a chiral center of convolutamydines. Additionally, the absolute configuration of natural convolutamydine B has been determined as R by its CD spectrum. PMID- 16468741 TI - DNA-protein cross-linking: model systems for pyrimidine-aromatic amino acid cross linking. AB - [reaction: see text] We have synthesized simple model systems to explore the possibility of photo-cross-linking between the pyrimidine bases and the side chains of the aromatic amino acids. Thymine/phenylalanine and thymine/tyrosine models gave cross-links, and thymine/tryptophan models gave complex mixtures; the cytosine/phenylalanine model was unreactive. The quantum yields for the model cross-linking reactions were 18-46 times smaller than those for thymine dimer formation. Biphotonic excitation contributes little to the yield of these reactions. PMID- 16468743 TI - A one-pot synthesis and functionalization of polyynes. AB - [reaction: see text] A one-pot synthesis and derivatization of diynes and triynes is reported. The polyyne framework is formed from a dibromoolefin precursor based on a carbenoid rearrangement, and the resulting Li-acetylide is then trapped in situ with an electrophile to provide functionalized di- and triynes. Alternatively, transmetalation of the Li-acetylide intermediate provides either the Zn- or Sn-acetylide, which then allows for the divergent preparation of diaryl polyynes or aryl ynones via palladium-catalyzed cross-coupling reactions. PMID- 16468742 TI - Design, total synthesis, and evaluation of novel open-chain epothilone analogues. AB - [structure: see text] The design, total synthesis, and biological evaluation of two open-chain analogues of epothilone incorporating the critical C1-C8 fragment and the aromatic side chain held together by a small molecular scaffold have been achieved. Biological evaluation revealed that further restraint between the flexible C1-C8 region and the molecular scaffold may be necessary for potent inhibition of cell proliferation. PMID- 16468744 TI - Gold(I)-catalyzed oxidative cleavage of a C-C double bond in water. AB - [reaction: see text] Oxidative cleavage of the C=C bond to afford ketone or aldehyde products with tert-butyl hydrogenperoxide (TBHP) as the oxidant can be catalyzed by AuCl with neocuproine (1) in water. PMID- 16468745 TI - Ring-opening dynamics of jadomycin A and B and dalomycin T. AB - [structure: see text] A novel oxazolone ring-opening and interconversion process between the two jadomycin diastereomeric forms has been characterized by NMR spectroscopy. An analogue, dalomycin T, has been isolated for the first time and does not undergo interconversion. PMID- 16468746 TI - Polyenylpyrroles and polyenylfurans from an Australian Isolate of the soil ascomycete Gymnoascus reessii. AB - [structure: see text] An Australian isolate of the soil ascomycete Gymnoascus reessii yielded a series of cytotoxic metabolites, including the known polyenylpyrroles rumbrin (1) and auxarconjugatin A (2), and the new rumbrin stereoisomer 12E-isorumbrin (3), as well as an unprecedented class of polyenylfurans exemplified by gymnoconjugatins A (4) and B (5). Structures were assigned with detailed spectroscopic analysis. PMID- 16468747 TI - Synthesis of bicyclo[3.1.0]hexanes functionalized at the tip of the cyclopropane ring. Application to the synthesis of carbocyclic nucleosides. AB - [reaction: see text] A general synthetic strategy for the preparation of functionalized bicyclo[3.1.0]hexanes is described. The new approach employs a cross metathesis step designed to functionalize the appropriate terminal olefin of the bicyclo[3.1.0]hexane precursor and a carbene-mediated intramolecular cyclopropanation reaction on the corresponding diazo intermediate. This combined methodology allowed the diastereoselective introduction of chemically diverse substituents at the tip of the cyclopropane group, except in cases where the substituents consisted of electron-withdrawing groups where a competing [3 + 2] cycloaddition predominated. PMID- 16468748 TI - Building addressable libraries: site selective coumarin synthesis and the "real time" signaling of antibody-coumarin binding. AB - [reaction: see text] The feasibility of using active semiconductor chips containing addressable arrays of microelectrodes for the "real-time" monitoring of biologically relevant binding events has been demonstrated by detecting the binding of a coumarin substrate by an anticoumarin antibody. The coumarin substrate was synthesized proximal to predetermined electrodes on the chip with the use of a Pd(II) reagent that was itself generated by using the selected electrodes. Once the coumarin was synthesized, its binding to the anticoumarin antibody was detected by monitoring the current associated with a ferrocene ferrocinium ion redox cycle that was established between the electrodes on the chip and a remote auxiliary electrode. PMID- 16468749 TI - Rapid assembly and in situ screening of bidentate inhibitors of protein tyrosine phosphatases. AB - [reaction: see text] We have successfully designed and synthesized a small library of protein tyrosine phosphatase (PTP) inhibitors, in which the so-called "click chemistry" or Cu(I)-catalyzed 1,3-dipolar alkyne-azide coupling reaction was carried out for rapid assembly of 66 different bidentate compounds. Subsequent in situ enzymatic screening revealed a potential PTP1B inhibitor (IC(50) = 4.7 microM) which is 10-100 fold more potent than other PTPs. PMID- 16468750 TI - New fluorophores with rod-shaped polycyano pi-conjugated structures: synthesis and photophysical properties. AB - [reaction: see text] Novel rod-shaped polycyano-oligo(phenyleneethynylene)s were synthesized by Pd cross-coupling reaction. Polycyano groups were found to greatly improve the emission efficiency (Phi(f)) of OPEs. By the end donor modification, we achieved the creation of very intense blue light-emitting fluorophore with the SMe group (Phi(f) = 0.972, log epsilon 4.89, lambda(em) 455 nm) and very intense yellow light-emitting fluorophore with the NMe(2) group (Phi(f) = 0.999, log epsilon 4.75, lambda(em) 555 nm). Contrasting Phi(f) solvent dependency of 6 and 7 and a linear relationship between Phi(f) and sigma(p)-X over the whole region of sigma(p)-X were also found. PMID- 16468751 TI - New ABC core for the synthesis of nonsymmetric star molecules. AB - [reaction: see text] An ABC core has been synthesized, with two orthogonal protecting groups (PG). This new multifunctional unit allows building of star shaped molecules with up to three different moieties. The synthesis of a star shaped mesogen based on an oligobenzoate scaffold with three arms of different lengths is demonstrated. PMID- 16468752 TI - Cobalt(II)-catalyzed cross-coupling between polyfunctional arylcopper reagents and aryl fluorides or tosylates. AB - [reaction: see text] Organocopper compounds prepared by the transmetalation of functionalized arylmagnesium halides with CuCN.2LiCl undergo smooth cross coupling reactions with aryl fluorides and tosylates bearing a carbonyl function in the ortho position in the presence of Co(acac)(2) (7.5 mol %), Bu(4)NI (1 equiv), and 4-fluorostyrene (20 mol %) as promoters in DME/THF/DMPU leading to polyfunctional aromatics or heterocycles. PMID- 16468754 TI - An iron-containing ionic liquid as recyclable catalyst for aryl grignard cross coupling of alkyl halides. AB - [reaction: see text] The ionic liquid butylmethylimidazolium tetrachloroferrate (bmim-FeCl(4)) was found to be a very effective and completely air stable catalyst for the biphasic Grignard cross-coupling with primary and secondary alkyl halides bearing beta-hydrogens. After simply decanting the product in the ethereal layer, the ionic liquid catalyst was successfully recycled four times. PMID- 16468753 TI - Palladium-catalyzed cross-coupling reactions of (2-pyridyl)allyldimethylsilanes with aryl iodides. AB - [reaction: see text] (2-Pyridyl)allyldimethylsilanes were found to be novel pyridyl transfer reagents in palladium-catalyzed cross-coupling reactions of various aryl iodides in the presence of silver(I) oxide as an activator. PMID- 16468755 TI - Potent algicides based on the cyanobacterial alkaloid nostocarboline. AB - [structure: see text] Nostocarboline and seven derivatives were prepared and displayed minimal inhibitory concentration (MIC) values >or=100 nM against the growth of Microcystis aeruginosa PCC 7806, Synechococcus PCC 6911, and Kirchneriella contorta SAG 11.81, probably via the inhibition of photosynthesis. The natural product hybrid nostocarboline/ciprofloxacin displayed additional antibacterial activity against several Gram-negative bacteria (MICs >or=0.7 microM). Nostocarboline can thus be considered a potent, selective, readily available, natural algicide. PMID- 16468756 TI - Hydrogen bond catalyzed direct reductive amination of ketones. AB - [reaction: see text] A novel, biomimetic concept for the direct reductive amination of ketones is described that relies on selective imine activation by hydrogen bond formation. The mild, acid- and metal-free process requires only catalytic amounts of thiourea as hydrogen bond donor and utilizes the Hantzsch ester for transfer hydrogenation. The method allows the efficient synthesis of structurally diverse amines. PMID- 16468758 TI - cis-endo-Bicyclo[1.1.1]pentane-1,2,3,4- tetracarboxylic acid and its derivatives. AB - [reaction: see text] We report the synthesis of cis-endo-bicyclo[1.1.1]pentane 1,2,3,4-tetracarboxylic acid and several of its derivatives with differentiated bridgehead and bridge positions, starting with tetracyclo[3.2.0.0(1,6).0(2,6)]heptane. PMID- 16468757 TI - Formal total synthesis of (-)-lepadiformine. AB - [reaction: see text] A stereocontrolled approach to the preparation of the Weinreb intermediate 3 has been developed. The important features of this approach are the creation of stereogenic centers through a cyclic amino acid ester-enolate Claisen rearrangement and the use of ring-closing metathesis for the construction of the azaspirocyclic skeleton. PMID- 16468759 TI - New terpenoids from Amentotaxus formosana. AB - [structure: see text] Amentoditaxone (1), possessing an unprecedented diterpenoid skeleton, along with two new terpenoids, amentotaxin WC (2) and amentotaxone (3), were established by extensive analysis of spectroscopic data. PMID- 16468760 TI - Synthesis of a peralkynylated pyrazino[2,3-g]quinoxaline. AB - [structure: see text] The synthesis of a hexaethynyl[2,3-g]pyrazinoquinoxaline and its crystal structure are reported. Starting from tetraaminobenzoquinone, condensation to bis(triisopropylsilyl)hexadiyne-2,3-dione affords 2,3,7,8 tetrakis(triisopropylsilylethynyl)pyrazino[2,3-g]quinoxaline-5,10-dione. Reaction with TIPS-CC-Li followed by reduction with hypophosphite in the presence of KI furnished the title molecule in a yield of 62%. Pd catalysis is not involved in any of these steps. PMID- 16468761 TI - Cage-shaped borate esters with enhanced Lewis acidity and catalytic activity. AB - [structure: see text] A cage shape causes high Lewis acidity and catalytic activity on boron. Borate esters that have cage-shaped ligands have accessible LUMO with lower eigenvalues than normal open-shaped borate esters. A large dihedral angle at C-O-B-O in cage-shaped borate esters induces less overlap between p-orbitals on O and B. The hetero-Diels-Alder reaction is effectively catalyzed by the cage-shaped borate, although the open-shaped borate does not act as a catalyst. PMID- 16468762 TI - Directed ortho-metalation of unprotected benzoic acids. Methodology and regioselective synthesis of useful contiguously 3- and 6-substituted 2 methoxybenzoic acid building blocks. AB - [reaction: see text] By treatment with s-BuLi/TMEDA at -78 degrees C, unprotected 2-methoxybenzoic acid is deprotonated exclusively in the position ortho to the carboxylate. A reversal of regioselectivity is observed when the acid is treated with n-BuLi/t-BuOK. These results are of general utility for the one-pot preparation of a variety of very simple 3- and 6-substituted 2-methoxybenzoic acids that are not easily accessible by conventional means. The potential usefulness of the method is demonstrated by the expedient synthesis of lunularic acid. PMID- 16468763 TI - A metallocene-pyrrolidinopyridine nucleophilic catalyst for asymmetric synthesis. AB - [reaction: see text] A highly active chiral 4-aminopyridine nucleophilic catalyst, available in three steps from (S,S)-hexane-2,5-diol, was applied to the asymmetric Steglich rearrangement of O-aceylated azlactones (1 mol % loading, up to 76% ee). PMID- 16468764 TI - Reagent-controlled asymmetric homologation of boronic esters by enantioenriched main-group chiral carbenoids. AB - [reaction: see text] Putative enantioenriched carbenoid species, (R)-1-chloro-2 phenylethylmagnesium chloride (9) and (S)-1-chloro-2-phenylethyllithium (26), generated in situ by sulfoxide ligand exchange from (-)-(R(S),R)-1-chloro-2 phenylethyl p-tolyl sulfoxide (8), effected the stereocontrolled homologation of boronic esters. sec-Alcohols derived from the product boronates by oxidation with basic hydrogen peroxide exhibited % ee closely approaching that of sulfoxide 8 in examples employing Li-carbenoid 26. PMID- 16468765 TI - A concise total synthesis of (-)-cylindricine C through a stereoselective intramolecular aza-[3 + 3] annulation strategy. AB - [reaction: see text] An enantioselective total synthesis of (-)-cylindricine C is described, featuring a diastereoselective intramolecular aza-[3 + 3] annulation strategy and an interesting halohydrin formation of the C4-5 olefin for construction of C4-ketone. This work provides a unique approach to this family of natural products. PMID- 16468766 TI - The Lewis acid-catalyzed nazarov reaction of 2-(n-methoxycarbonylamino)-1,4 pentadien-3-ones. AB - [reaction: see text] A highly efficient carbonylative Suzuki-Miyaura coupling reaction of lactam-derived vinyl triflates and alkenylboronic acids afforded 2-(N methoxycarbonylamino)-1,4-pentadien-3-ones as suitable substrates for the Nazarov reaction. The most competent Lewis acids for the Nazarov reaction were Cu(OTf)(2) (2 mol %) and Sc(OTf)(3) (3 mol %) in DCE, which provided the Nazarov products in excellent yield. As both the carbonylative coupling and the subsequent Nazarov reaction were high yielding, the overall methodology is a concise and efficient route to [1]pyrindine systems. PMID- 16468767 TI - Synthesis and characterization of soluble oligo (9,10-bisalkynylanthrylene)s. AB - [structure: see text] Soluble oligo(9,10-bialkynylanthrylene)s up to pentamers were synthesized by means of the Suzuki coupling reaction. A solution processed thin film field-effect transistor from pentamer OA-5b shows a charge carrier mobility of 2.95 x 10(-)(3) cm(2)/V x s. PMID- 16468768 TI - Stereoselective synthesis of beta-substituted beta-amino sulfones and sulfonamides via addition of sulfonyl anions to chiral N-sulfinyl imines. AB - [reaction: see text] A highly stereoselective synthesis of beta-amino sulfones and sulfonamides via addition of sulfonyl anions to chiral N-sulfinyl imines is described. The addition reaction proceeds in good yield (75-99%) and stereoselectivity. PMID- 16468769 TI - Palladium-catalyzed cross-coupling reactions of heterocyclic silanolates with substituted aryl iodides and bromides. AB - [reaction: see text] Sodium silanolates derived from a number of heterocyclic silanols undergo cross-coupling with a variety of aromatic iodides and bromides under mild conditions. In situ deprotonation of the silanols with an equivalent amount of sodium hydride in toluene generates the sodium salt that couples with iodides under the action of Pd(2)(dba)(3).CHCl(3) in good yield at room temperature to 50 degrees C. The aromatic bromides also couple with these salts under the action of the Pd(I) catalyst 12. PMID- 16468770 TI - Design and synthesis of a potent phorboxazole C(11-15) acetal analogue. AB - [structure: see text] We disclose here the design, synthesis, and biological evaluation of simplified Z- and E-C(2-3) alkynyl phorboxazole C(11-15) acetals (+)-7Z and (+)-7E, wherein the Z-isomer proved to be a potent nanomolar cytotoxic agent. Reevaluation of (+)-C(45-46) E-chloroalkenyl phorboxazole A (6) confirms subnanomolar activity across a broad panel of human cancer cell lines. PMID- 16468774 TI - Directed ortho metallation of deep cavity cavitands: functionalizing molecular concavity. AB - Directed ortho metallation (DOM) processes have been used to functionalize the cavity and rim of title cavitand 1. The preorganization of the host resulted in a considerable reduction in the range of products produced. Thus, whereas sixty nine products are possible from per-functionalization, only twelve were observed when the host was treated with three different alkyllithiums. PMID- 16468775 TI - N-nitroso- and N-nitraminotetrazoles. AB - N-Nitroso- (5a,c) and N-nitraminotetrazoles (6a-c) were synthesized from the corresponding aminotetrazoles (3a-c) either by the direct nitration with acetic anhydride/HNO3 or by dehydration of the corresponding nitrates (4a-c) with concentrated sulfuric acid. The conversion of the N-nitrosoaminotetrazoles (5a,c) with peroxytrifluoroacetic acid (CF3CO3H) yielded the corresponding nitramines in high yield (6a (82%), 6c (80%)). The N-nitroso- (5a,c) and N-nitraminotetrazoles (6a-c) have been fully characterized by vibrational (IR, Raman) and multinuclear NMR spectroscopy (14N/15N, 1H, 13C), mass spectrometry, and elemental analysis. A detailed discussion of the 15N chemical shifts and 1H-15N coupling constants is given. The molecular structures in the solid state were determined by single crystal X-ray diffraction (3a,c; 5a,c; 6a-c) and a detailed discussion of the molecular structures will be presented. Furthermore, the structure and bonding as well as N,N rotational barriers are discussed on the basis of theoretically obtained data (B3LYP/6-31G(d,p), NBO analysis). In the case of two N nitraminotetrazoles (6a,c) the physicochemical properties (e.g., D, P, delta(f)H degrees) were evaluated. The heat of formation was calculated to be positive for 6a and 6c (+2.8 and +85.2 kcal mol(-1), respectively) and the calculated detonation velocity with 5988 (6a) and 7181 (6c) m s(-1) reaches values of TNT and nitroglycerin. PMID- 16468777 TI - Zn(pybox)-complex-catalyzed asymmetric aqueous Mukaiyama-aldol reactions. AB - Catalytic asymmetric aldol reactions in aqueous media have been developed using chiral zinc complex. The aldol products have been obtained in high yields, high diastereocontrol, and good level of enantioselectivity. Various aromatic and alpha,beta-unsaturated aldehydes and silyl enol ethers derived from ketones can be employed in this reaction to provide the aldol adducts in good to high yield. The elaborated catalytic system has been found as selective for aliphatic aldehydes as well. PMID- 16468776 TI - Synthesis and properties of trans-3',4'-bridged nucleic acids having typical S type sugar conformation. AB - The synthesis of nucleoside analogues with a conformationally restricted sugar moiety is of great interest. The present research describes the synthesis of BNA (bridged nucleic acid) monomers 1 and 2 bearing a 4,7-dioxabicyclo[4.3.0]nonane skeleton and a methoxy group at the C2' position. Conformational analysis showed that the sugar moiety of these monomers is restricted in a typical S-type conformation. It was difficult to synthesize the phosphoramidite derivative of the ribo-type monomer 1, while the phosphoramidite of the arabino-type monomer 2 was successfully prepared and incorporated into oligodeoxynucleotides (ODNs). The hybridization ability of the obtained ODN derivatives containing 2 with complementary strands was evaluated by melting temperature (T(m)) measurements. As a result, the ODN derivatives hybridized with DNA and RNA complements in a sequence-selective manner, though the stability of the duplexes was lower than that of the corresponding natural DNA/DNA or DNA/RNA duplex. PMID- 16468778 TI - An improved aldehyde linker for the solid phase synthesis of hindered amides. AB - A novel aldehyde dual-linker system has been developed for the solid phase synthesis of sterically hindered amides. The linker [5-(4-formyl-3 hydroxyphenoxy)pentanoic acid] exploits an intramolecular oxygen-nitrogen acyl transfer mechanism to prepare compounds that are unattainable with current commercially available linkers. A dual linker system, exploiting the hyper-acid labile Sieber amide linker as part of the construct, enabled the initial reductive alkylation reactions of hindered amines and their subsequent acylation with a range of carboxylic acids with varying stereoelectronic properties to be monitored. Simple acylation conditions (HBTU/HOBt/NMM) sufficed to provide near quantitative reaction of test acids with support-bound hindered amines, reaction conditions which failed when commercial linkers were used. PMID- 16468779 TI - Synthesis of isoprostanyl phosphatidylcholine and isoprostanyl phosphatidylethanolamine. AB - The syntheses of two isoprostanyl phospholipids are described. A newly established route to 15-F(2t)-isoprostane and ent-15-epi-F(2t)-isoprostane has allowed for the selective preparation of 15-F(2t)-isoprostanyl phosphatidylethanolamine and ent-15-epi-F(2t)-isoprostanyl phosphatidylcholine. The nature of the headgroups dictates the coupling strategy used to attach the appropriately protected isoprostanes to the corresponding lysophospholipids. Preliminary 1H NMR and 31P NMR studies indicate that these isoprostanyl phospholipids aggregate in apolar solvents. PMID- 16468780 TI - A flexible stereospecific synthesis of polyhydroxylated pyrrolizidines from commercially available pyranosides. AB - Nitrogen-containing sugar analogues, known as azasugars or iminosugars, such as polyhydroxylated piperdines, pyrrolidines, pyrrolizidines, and indolizidines, have the potential to become important therapeutic agents due to their ability to inhibit glycosidases. Synthetic pathways that are able to systematically produce a variety of these azasugars are eagerly sought after, since even minute structural or stereochemical changes often significantly alter the degree of inhibition. The synthesis of tetrahydroxylated pyrrolizidines 40 and 41 starting from methyl alpha-d-glucopyranoside is described and will be used as a template to develop syntheses of all the stereoisomers of polyhydroxylated pyrrolizidine 9 as well as other analogous bicyclic polyhydroxylated iminosugars. The key steps in this synthesis involve a one-pot conversion of a halopyranoside to a divinylamine by employing a simultaneous Zn reduction and reductive amination of the resulting aldehyde. After protection of the amine, a ring-closing metathesis results in a multifunctional eight-membered ring that then undergoes an internal S(N)2 cyclization to form an alkene-containing pyrrolizidine 33. Dihydroxylation of the alkene followed by hydrogenolysis of the benzyl protecting groups results in tetrahydroxylated pyrrolizidines 40 and 41. PMID- 16468781 TI - Substituent-dependent anomeric effects as a source of conformational preference in pyridinium methylides. AB - A systematic density functional theory level investigation of differently substituted pyridinium methylides was carried out to determine the role of C(ylidic) lone-pair-associated hyperconjugative and negative hyperconjugative interactions in deciding conformational preferences. Deviation from the coplanar orientation of the carbanionic center with the pyridine ring and its substituent dependence has been found to correlate well with the relative opportunities for conjugative and negative hyperconjugative interactions of a ylidic moiety with different substituent groups present at the ylidic carbon. The contribution of individual n-->pi* conjugative, n-->sigma* negative hyperconjugative, and sigma- >pi* hyperconjugative interactions in a particular conformation of pyridinium dichlorophosphinomethylides was assessed from donor-acceptor stabilization energies, as obtained from natural bond orbital (NBO) analysis. The relative extent of conjugative and negative hyperconjugative interactions with the substituents present at the ylidic carbon plays an important role in permitting the delocalization of ylidic charge into the pyridine ring, thereby controlling the relative orientation of the latter with the carbanionic plane. PMID- 16468782 TI - Facile synthesis and characterization of naphthidines as a new class of highly nonplanar electron donors giving robust radical cations. AB - Naphthidines 2 were prepared by nickel-catalyzed amination of 1-chloronaphthalene followed by oxidative homocoupling of 1-naphthalene amines 1 using titanium(IV) tetrachloride. The electronic and magnetic properties of materials 2 were investigated by cyclic voltammetry and other electrochemical techniques, EPR and UV-visible spectroscopies, and magnetic susceptibility. It was demonstrated that compounds 2 could be easily and reversibly oxidized via a two-electron-transfer reaction into their bis(radical cation) 2(2.2+), which displays a substantial stability at room temperature (the half-life of 2(2.2+) estimated by EPR at 25 degrees C was 10 days). B3LYP/6-31G optimized structures of N,N'-bis(4 methoxyphenyl)-(1,1'-binaphthyl)-4,4'-diamine 2g shows significant differences in the torsion angle between the naphthalene moieties depending on its oxidation state. Twisted structures are preferred for neutral compounds, whereas more planar are favored for the oxidized forms 2g*+ and 2g(2.2+) to realize spin and/or charge delocalizations over the whole pi-system. Such conformation changes concerted with the electron transfers contribute to explain the unusual two electron process observed in the electrochemical behavior of 2g instead of the two single-electron transfers that would have been expected in the case of two successive oxidations. It is finally shown that the oxidation of 2g in CH2Cl2 with thianthrenium perchlorate (ThClO4) generates the dication 2g(2.2+) with singlet spin-multiplicity. PMID- 16468783 TI - Rigid molecular tripod with an adamantane framework and thiol legs. Synthesis and observation of an ordered monolayer on Au111. AB - Tripod-shaped trithiols 1-3, containing CH2SH groups at the three bridgehead positions of the adamantane framework and a halogen-containing group [Br (1), p BrC6H4 (2), or p-IC6H4 (3)] at the fourth bridgehead, were synthesized, and self assembled monolayers (SAMs) were prepared on atomically flat Au111 surfaces. The three-point chemisorption of these tripods was confirmed by polarization modulation infrared reflection absorption spectroscopy, which showed the absence of a S-H stretching band. Scanning tunneling microscopy of the SAM of 1 exhibited a hexagonal arrangement of the adsorbed molecule with a lattice constant of 8.7 angstroms. A unidirectionally oriented, head-to-tail array of 1, which allows the close approach of neighboring molecules, is proposed as a reasonable model of the two-dimensional crystal, where the adsorbed sulfur atoms form a quasi-(radical3 x radical3)R30 degrees lattice. The charge of the electrochemical reductive desorption of the SAM of 1 was in good agreement with the expected surface coverage, while the SAMs of 2 and 3 showed somewhat less (ca. 70%) charge. The large negative reduction peak potentials, observed for the SAM of 1, are taken to indicate a tight anchoring of this tripod by three sulfur atoms. PMID- 16468784 TI - Total synthesis of 8,12-iso-iPF3alpha-VI, an EPA-derived isoprostane: stereoselective introduction of the fifth asymmetric center. AB - [reaction: see text] A new and stereoselective approach for the synthesis of all syn isoprostanes is reported. This method, which is based on acid-catalyzed Diels Alder reaction, allows the introduction of the side chain with a predetermined stereochemistry of the hydroxy group. The first total synthesis of an eicosapentaenoic acid (EPA)-derived iP, 8,12-iso-iPF3alpha-VI 10, was performed using this approach. PMID- 16468785 TI - New glycomimetics: anomeric sulfonates, sulfenamides, and sulfonamides. AB - The synthesis of a variety of new 1-thio-D-glucopyranose derivatives oxidized at the sulfur atom is described, including seven 1-C-sulfonic acids, three sulfonate esters, three sulfinate esters, an S,S'-diglycosyl thiolsulfonate and thiolsulfinate, four S-glycosyl sulfenamides, an S-glycosyl sulfinamide, and two S-glycosyl sulfonamides. These compounds possess unusual anomeric functionality that might be resistant or even inhibitory to normal enzymatic carbohydrate processing, and therefore, they may be of future use in studies of enzyme inhibition, structure, mechanism, and function. PMID- 16468786 TI - Synthesis of a nonavalent mannoside glycodendrimer based on pentaerythritol. AB - A nonavalent glycodendrimer bearing terminal alpha-d-mannopyranoside units has been synthesized with a convergent approach. Terminal trivalent mannoside dendrons bearing p-halophenyl ethers were prepared by glycosylation of pentaerythritol derivatives having three 2-hydroxyethyl ether substituents. Two efficient routes were developed for the synthesis of the pentaerythritol-based core (17), which has three terminal propargyl ethers. Conditions were found under which the triple Sonogashira coupling reaction of the dendron and the tri-O propargyl ether (17) proceeded efficiently. The product was deprotected and it and precursors were fully characterized by NMR spectroscopy and FT-ICR mass spectrometry. PMID- 16468787 TI - Synthesis of azolyl carboximidamides as ligands for Zn(II) and Cu(II): application of the Zn(II) complexes as catalysts for the copolymerization of carbon dioxide and epoxides. AB - A series of novel S,N-heterocyclic (thiazolyl) substituted carboximidamides 3 and 4 was synthesized in yields up to 82% from specific triazinium salts 1 and primary or secondary amines 2 which additionally bear pyridine or imidazole units. These carboximidamides are used as tailor-made ligands for the complexation of Cu(II) and Zn(II). The coordination behavior of 3 and 4 and the properties of the resulting metal complexes are affected a significant extent especially by the nature of these amine substituents. The most important structural feature of the novel complexes is that the ligation of the metal cations is achieved by a 1,3,5-triazapentadienyl anion system, compare the X-ray structure of the model complex Cu-4d. Analogous Zn(II) complexes 5, 6a, 6b, 6c, 7a, and 7b were obtained from carboximidamides 3, 4a, 4b, 4c, 4d, and 4e after reaction with diethylzinc. Interestingly, these Zn(II) complexes possess an intrinsic activity to catalyze the copolymerization of cyclohexene oxide and carbon dioxide to give polycarbonates 15 (TON up to 113; Turn Over Number: moles of substrate 14 consumed per moles of zinc. Molecular weights: up to 206.10(3) Da). Contaminations of 15 by polyethers are produced only in remarkably small amounts. PMID- 16468788 TI - Unprecedented lability of the 5'-O-tert-butyldimethylsilyl group from 3'-spiro 5''-(4''-acylamino-1'',2''-oxathiole-2'',2''-dioxide) nucleoside derivatives via neighboring group participation of the 4''-acylamino residue. AB - Scarce examples of exceptionally mild desilylation of tert-butyldimethylsilyl (TBDMS) ether groups by neighboring group participation have been previously described. Here, we investigate, in detail, the discovery of the unusual lability of the 5'-TBDMS group on 4' '-acylamino TSAO derivatives in DMSO solution. The synthesis and comparative chemical stability studies in different solvents of a variety of 4' '-substituted TSAO derivatives bearing different carbonyl functionalities are reported. Modifications have also been performed at the 5' position of the TSAO molecule to gain insight into the structural requirements for the desilylation to occur. The role of the solvent has also been studied. Additionally, NMR and theoretical investigations have been carried out to get further insight into the conformational, geometric, and/or electronic parameters that may play a role in the "spontaneous" release of the 5'-TBDMS group. A silyl hydrolysis mechanism involving neighboring group participation of the 4' ' acylamino group is proposed. PMID- 16468789 TI - Synthesis of the proposed structure of mucoxin via regio- and stereoselective tetrahydrofuran ring-forming strategies. AB - An enantioselective total synthesis of the proposed structure of mucoxin (1) is described. Mucoxin, an annonaceous acetogenin isolated from bioactive leaf extracts of Rollinia mucosa, is the first acetogenin containing a hydroxylated trisubstituted tetrahydrofuran (THF) ring. This natural product is a highly potent and specific antitumor agent against MCF-7 (breast carcinoma) cell lines (ED50 = 3.7 x 10(-3) microg/mL compared to adriamycin, ED50 = 1.0 x 10(-2) microg/mL). The total synthesis described herein features two regio- and stereoselective THF ring-forming reactions. The 2,3,5-trisubstituted THF portion (C13-C17) was accessed using a highly regioselective cyclization of a methylene interrupted epoxydiol, and the 2,5-disubstituted THF ring (C8-C12) was conveniently assembled via a 1,2-n-triol cyclization strategy. The spectral data of the synthetic material and two of its diastereomers did not match the reported data for the natural product. On the basis of detailed spectroscopic analysis of the synthesized molecule, we reason that the spectral discrepancies are due to stereochemical misassignment of the natural product. PMID- 16468791 TI - Electrolytic partial fluorination of organic compounds. 83. Anodic fluorination of N-substituted pyrroles and its synthetic applications to gem-difluorinated heterocyclic compounds. AB - Anodic fluorination of N-substituted pyrroles was carried out in MeCN containing supporting fluoride salts such as Et3N-nHF (n = 2-5) and Et4NF-4HF with use of platinum electrodes under constant current conditions. Anodic fluorination of N methyl- and N-p-tosylpyrroles having an electron-withdrawing cyano group proceeded smoothly to provide the corresponding fluorinated products in moderate to excellent yields, while anodic fluorination of N-methylpyrrole devoid of an electron-withdrawing cyano group did not take place and a polymeric product was formed at the anode surface. In sharp contrast to the cases of N-methylpyrroles, even N-p-tosylpyrrole devoid of an electron-withdrawing cyano group underwent anodic fluorination efficiently. Diels-Alder reaction of 5,5-difluoro-1-methyl-3 pyrrolin-2-one (2e) derived from anodic fluorination of 2-cyano-1-methylpyrrole (2a) with various dienes was carried out to provide the cycloaddition products in excellent yields. Furthermore, Michael reaction of 2e with various nucleophiles was also successfully carried out to provide the Michael addition products in good to excellent yields. PMID- 16468790 TI - C-metalated nitriles: electrophile-dependent alkylations and acylations. AB - Sequential carbonyl addition-conjugate addition of Grignard reagents to 3 oxocyclohex-1-ene-1-carbonitrile generates C-magnesiated nitriles whose alkylation stereoselectivities intimately depend on the nature of the electrophile. The alkylation of these C-magnesiated nitriles with alkyl halides, sulfonates, and unstrained ketones occurs with the retention of the C-Mg configuration, whereas aldehyde and acyl cyanide acylations proceed with inversion of the stereochemistry. Mechanistic probes indicate that the stereoselectivity is controlled by stereoelectronic effects for most electrophiles, except allylic, benzylic, and cyclopropyl halides where single electron-transfer processes intervene. Screening numerous alkylations of C magnesiated nitriles with a diverse range of electrophiles reveals the reaction scope and delineates the fundamental stereoelectronic effects responsible for the highly unusual electrophile-dependent alkylations. PMID- 16468792 TI - Synthesis of highly substituted enantiopure piperazines and ketopiperazines from vicinal N-sulfinyl diamines. AB - Enantiopure 1-benzyl-2,3-disubstituted piperazines (4) have been synthesized by treatment of N-sulfinyl-N-benzyldiamino alcohols (1) with diethyl oxalate and sodium methoxide followed by reduction with borane. Alternatively, the sulfinamido group was preserved by an N-acylation/cyclization protocol using alpha-chloroacetyl chloride that led to the synthesis of N-sulfinyl ketopiperazines (11). Ensuing elimination of the sulfinyl group with NaH produced imino ketopiperazines (9) that are suitably functionalized for nucleophilic addition to the imino moiety. Stereoselective and high yielding allylation of imino ketopiperazines (9c) was achieved under Barbier conditions using CeCl3.7H2O as the additive. PMID- 16468794 TI - Structurally simple pyridine N-oxides as efficient organocatalysts for the enantioselective allylation of aromatic aldehydes. AB - A series of structurally simple pyridine N-oxides have readily been assembled from inexpensive amino acids and tested as organocatalysts in the allylation of aldehydes with allyl(trichloro)silane to afford homoallylic alcohols. (S)-proline based catalysts afforded the products derived from aromatic aldehydes in fair to good yields and in up to 84% enantiomeric excess (ee). The allylation of heteroaromatic, unsaturated, and aliphatic aldehydes was less satisfactory. By running the reaction in the presence of achiral and chiral additives and structurally different catalysts, we collected some insights into the relationship between the stereochemical outcome and the catalyst's structural features. Even if the ee's obtained are inferior to the best values observed with other catalysts, this work concurs to show that structurally simple pyridine N oxides can also promote the allylation reaction with satisfactory stereocontrol. PMID- 16468795 TI - Highly chemoselective methylation and esterification reactions with dimethyl carbonate in the presence of NaY faujasite. The case of mercaptophenols, mercaptobenzoic acids, and carboxylic acids bearing OH substituents. AB - In the presence of NaY faujasite, the reactions of dimethyl carbonate (DMC) with several ambident nucleophiles such as o- and p-mercaptophenols (1a,b), o- and p mercaptobenzoic acids (2a,b), o- and p-hydroxybenzoic acids (3a,b), mandelic and phenyllactic acids (4, 5), have been explored under batch conditions. Highly chemoselective reactions can be performed: at 150 degrees C, compounds 1 and 2 undergo only a S-methylation reaction, without affecting OH and CO2H groups; at 165 degrees C, acids 3-5 form the corresponding methyl esters, while both their aromatic and aliphatic OH substituents are fully preserved from methylation and/or transesterification processes. Typical selectivities are of 90-98% and isolated yields of products (S-methyl derivatives and methyl esters, respectively) are in the range of 85-96%. A comparative study with K2CO3 as a catalyst is also reported. Although the base (K2CO3) turns out to be more active than the zeolite, the chemoselectivity is elusive: compounds 2a,b undergo simultaneous S-methylation and esterification reactions, and acids 3-5 yield complex mixtures of products of O-methylation, O-methoxycarbonylation, and esterification of their OH and CO2H groups, respectively. Overall, the combined use of a nontoxic reagent/solvent (DMC) and a safe promoter (NaY) imparts a genuine ecofriendly nature to the investigated synthesis. PMID- 16468793 TI - Synthesis of new macrocyclic chiral manganese(III) Schiff bases as catalysts for asymmetric epoxidation. AB - We describe a general synthetic strategy for the preparation of a series of macrocyclic chiral manganese(III) salen complexes. The developed reaction pathway allows the modulation of the different key groups, namely, the chiral diimine, the bulky substituents in positions 3 and 3', and the linker used in the macrocyclization of the Schiff base. The different complexes presented here illustrate these readily available structural variations. The catalytic properties of the catalysts (5 mol %) were improved for the asymmetric epoxidation of 2,2'-dimethylchromene with NaOCl or H2O2 as oxygen atom donor. A large range of enantiomeric excesses was obtained (ee values from 30% to 96%), depending on the features and the stability of the complexes. The most efficient catalyst, in terms of stereoinduction (ee value = 96%), contains a diiminocyclohexyl moiety, ethyl groups in positions 3 and 3', and a short polyether junction arm. PMID- 16468796 TI - Solid-supported copper catalysts for atom-transfer radical cyclizations: assessment of support type and ligand structure on catalyst performance in the synthesis of nitrogen heterocycles. AB - A range of solid supported pyridinemethanimine 9-11 and polyamine 12-15 ligands have been prepared on silica, polystyrene, and JandaJel supports. The CuCl and CuBr complexes of these supported ligands have been used to assess both the effect of the ligand type and the nature of the support upon a representative range of copper-mediated atom transfer 5-exo-trig 6, 24-25, 5-exo-dig 26, 4-exo trig 28, and 5-endo-trig 27, 38 radical cyclizations to give nitrogen heterocycles. In addition, the effect of the nature of the support on the stereochemical outcome of the 5-exo cyclization of 25 has been probed. Generally, it was found that the type of support (e.g., polystyrene, silica, or JandaJel) had very little effect upon the efficiency and selectivity of the processes but that the nature of the ligand type immobilized was the important factor. Thus, the 5-exo cyclization of 6 and 24-26 proceeded more rapidly with the PMI ligands 9-11, whereas 4-exo cyclizations 28 and 5-endo radical polar crossover reactions 27 and 38 proceeded more efficiently with the JJ-TEDETA ligand 15. The efficiency of the supported ligands was also compared to their solution counterparts 4 and 5. The reusability of P-PMDETA ligand system 13 was assessed in the cyclization of 6. PMID- 16468797 TI - Methoxy-substituted stilbenes, styrenes, and 1-arylpropenes: photophysical properties and photoadditions of alcohols. AB - The photochemistry of trans-stilbene and four methoxy-substituted stilbene derivatives has been investigated in a variety of solvents. The fluorescence of all five trans isomers was quenched by 2,2,2-trifluoroethanol (TFE). Upon irradiation of the five substrates in TFE, the products derived from photoaddition of the solvent were detected. Nuclear magnetic resonance spectroscopy of the products formed by irradiation in TFE-OD indicated that the proton and nucleophile are attached to two adjacent atoms of the original alkene double bond. Irradiation of the corresponding methoxy-substituted styrenes and trans-1-arylpropenes in TFE produced the analogous solvent adducts. The photoaddition of TFE proceeded with the general order of reactivity: styrenes > trans-1-arylpropenes > trans-stilbenes. Transient carbocation intermediates were observed following laser flash photolysis of the stilbenes in 1,1,1,3,3,3 hexafluoro-2-propanol (HFIP). The results are consistent with a mechanism that involves photoprotonation of the substrates by TFE or HFIP, followed by nucleophilic trapping of short-lived carbocation intermediates. Compared to the other stilbene derivatives, trans-3,5-dimethoxystilbene displayed a large quantum yield of fluorescence and a low quantum yield of trans-cis isomerization in polar organic solvents. The unique photophysical properties of trans-3,5 dimethoxystilbene are attributed to formation of a highly polarized charge transfer excited state (mu(e) = 13.2 D). PMID- 16468799 TI - The scope and limitations of 1,3-stannyl shift-promoted intramolecular cyclizations of alpha-stannyl radicals with a formyl group. AB - Alpha-tributylstannyl radicals can be generated from the corresponding bromides or xanthates. These radicals undergo efficient intramolecular 1,5-cyclizations with a formyl group. The resulting beta-stannyl alkoxy radicals proceed through a 1,3-stannyl shift from carbon to oxygen to afford beta-stannyloxy radicals. This novel rearrangement is most likely irreversible and serves as a driving force to promote the cyclizations. Although the cyclization rates can be accelerated when the formyl group carries alpha-dimethyl substituents, unfortunately beta-scission of the alkoxy radicals becomes competitive with the 1,3-stannyl shift. The beta stannyloxy radicals can be employed in further cyclizations to obtain tandem cyclization products. PMID- 16468798 TI - Syn- and anti-selective Prins cyclizations of delta,epsilon-unsaturated ketones to 1,3-halohydrins with Lewis acids. AB - Ten acyclic and monocyclic delta,epsilon-unsaturated ketones, with and without methyl substituents on the double bond, underwent halide-terminated Prins (halo Prins) cyclizations under anhydrous conditions in the presence of Lewis acids. TiCl4, TiBr4, BCl3, and BBr3 promoted syn-selective cyclizations to sterically congested chloro- and bromohydrins, while SnCl4, SnBr4, InCl3, ZrCl4, and several other Lewis acids effected highly anti-selective reactions to furnish the corresponding trans halohydrins. The stronger Lewis acids (TiX4 and BX3) favor the syn process that involves axial delivery of a halide ligand. Competition experiments showed that substitution at the delta carbon (methallyl enones) led to increased rates (40-50-fold), while substitution at the epsilon position (cis and trans crotyl enones) retarded the rate and eroded the selectivity of the cyclizations. The trends in syn vs anti selectivity, reactivity, and effects of different Lewis acidic metal halides are rationalized by competitive reaction pathways proceeding through syn carbocation-halide ion pairs and a higher order transition state that leads to inversion of configuration and formation of trans halohydrins, along with cyclic olefins arising from proton elimination. PMID- 16468800 TI - Chiral phosphoramide-catalyzed enantioselective addition of allylic trichlorosilanes to aldehydes. Preparative and mechanistic studies with monodentate phosphorus-based amides. AB - The addition of allylic trichlorosilanes to benzaldehyde promoted by chiral phosphoramides to give the enantioenriched homoallylic alcohol has been investigated. In a survey of Lewis bases as activators for the addition of allyltrichlorosilane to benzaldehyde, phosphorus-based amides have been found to be the most effective promoters. To achieve asymmetric induction, chiral phosphoric triamides derived from chiral diamines have been developed and applied in the allylation reaction albeit with modest enantioselectivities. The addition of 2-butenylsilanes was highly diastereoselective, suggesting a closed, chair like transition structure. A detailed mechanistic study has been carried out to probe into the origin of activation. From a combination of nonlinear effects and kinetics studies, the reaction was found to likely involve two phosphoramides in both the rate and stereochemistry determining steps. These studies provided the background for the development of highly selective and reactive catalysts. PMID- 16468801 TI - Chiral phosphoramide-catalyzed enantioselective addition of allylic trichlorosilanes to aldehydes. Preparative studies with bidentate phosphorus based amides. AB - On the basis of the mechanistic insight that more than one Lewis basic moiety (phosphoramide) is involved in the rate- and stereochemistry-determining step of enantioselective allylation, bidentate chiral phosphoramides were developed. Different chiral phosphoramide moieties were connected by tethers of methylene chains of varying length. The rate and enantioselectivity of allylation with allyltrichlorosilane promoted by the bidentate phosphoramides was found to be highly dependent on the tether length. A new phosphoramide based on a 2,2' bispyrrolidine skeleton has been designed and afforded good yield, efficient turnover, and high enantioselectivity in allylation reactions. The synthesis of enantiopure 2,2'-bispyrrolidine was easily accomplished on large scale by photodimerization of pyrrolidine followed by resolution with L(or D)-tartaric acid. The scope of the allylation reaction was examined with variously substituted allylic trichlorosilanes and unsaturated aldehydes. This method has been applied to the construction of stereogenic, quaternary centers by the addition of unsymmetrically gamma-disubstituted allylic trichlorosilanes. PMID- 16468802 TI - Boron trifluoride-induced, new stereospecific rearrangements of chiral epoxy ethers. Ready access to enantiopure 4-(diarylmethyl)-1,3-dioxolanes and 4,5 disubstituted tetrahydrobenzo[c]oxepin-4-ols. AB - Upon treatment with BF3.Et2O at low temperature, enantiopure benzyl-type ethers of arylglycidols with electron withdrawing substituents at the skeletal aryl group and electron donating substituents at the benzyl group undergo stereospecific rearrangements of Friedel-Crafts type, leading to enantiopure 4 diarylmethyl-1,3-dioxolanes (2) or to enantiopure trans-4,5 disubstituted tetrahydrobenzo[c]oxepin-4-ols (5). The course of the reactions is controlled by the substitution pattern at the benzyl ether: While benzylic systems activated toward ipso substitution afford diarylmethanes 2 through a Friedel-Crafts reaction followed by fragmentation, benzylic systems activated toward ortho attack lead to enantiopure oxepinols 5 through a 7-endo-tet ring closure of Friedel-Crafts type. PMID- 16468803 TI - Tether-directed selective synthesis of fulleropyrrolidine bisadducts. AB - Selective synthesis of C60 bisadducts has been achieved by using the Prato 1,3 dipolar cycloaddition of tethered bis-azomethine ylides. New bis(benzaldehydes) 1 4 tethered by a rigid linker were prepared and used to direct the second cycloaddition of azomethine ylide to C60. Equatorial, trans-4, trans-3, trans-2, and trans-1 bisadducts have been selectively prepared with this approach. However, the introduction of chiral centers in the pyrrolidine rings in the course of the reaction complicated the chemistry, as a number of stereoisomers theoretically could be formed. The structure determination of the isomeric bisadducts was made based on spectroscopic data and theoretical calculations. To our best knowledge, this represents the first example of a systematic study on tether-directed selective synthesis of C60 fulleropyrrolidine bisadducts. PMID- 16468804 TI - Copper-promoted carbon-nitrogen bond formation with 2-iodo-selenophene and amides. AB - We present here carbon-nitrogen bond formation via a coupling reaction of 2-iodo selenophene catalyzed by Cu(I) in the presence of a base and an inexpensive ligand, and establish the first route to obtaining 2-nitrogen-selenophene derivatives in good yields. We can anticipate that this reaction works well with oxazolidinones, lactams, and aliphatic and aromatic amides, as nitrogen sources, in the absence of any supplementary additives. In addition, the reaction proceeded cleanly under mild reaction conditions and was sensitive to the ratio of amide/2-iodo-selenophene, as well as the nature of the ligand, base, and solvent. PMID- 16468805 TI - Dimethylzinc-mediated alkynylation of imines. AB - The treatment of various aromatic and aliphatic aldimines with a mixture of a terminal alkyne and a commercially available dimethylzinc solution in toluene yields the corresponding protected propargylic amines in moderate to excellent yields. The reaction proceeds in the absence of any activator. These observations led to the development of a three-component synthesis of propargylic amines in which the product was obtained upon mixing an aldehyde with ortho-methoxyaniline and phenylacetylene in the presence of dimethylzinc, through in situ formation of the corresponding imine. PMID- 16468807 TI - Metal-catalyzed oxidation and epoxidation of alpha-hydroxy vinyl and dienyl sulfoxides. AB - Treatment of acyclic alpha-hydroxyalkyl alpha,beta-unsaturated sulfoxides with t BuOOH/VO(acac)2 results in rapid oxidation to the unsaturated sulfones followed by an unusual regio- and stereoselective epoxidation at the unsaturated sulfones; this methodology has been applied to the preparation of carbohydrate-like fragments. PMID- 16468806 TI - Palladium(0)-catalyzed synthesis of chiral ene-allenes using alkenyl trifluoroborates. AB - Enantioenriched allenes serve as chiral transfer reagents, making them attractive synthetic targets. Herein, the synthesis of enantioenriched allenes utilizing a Pd(0)-catalyzed cross-coupling reaction of propargylic carbonates and phosphates with alkenyl trifluoroborates is reported. Di-, tri-, and tetrasubstituted allenes were synthesized in moderate to high optical yields. Several racemic allenes possessing various functional groups were also synthesized. PMID- 16468808 TI - Estimating the limiting reducing power of SmI2/H2O/amine and YbI2/H2O/amine by efficient reduction of unsaturated hydrocarbons. AB - The mixture of samarium diiodide, amine, and water (SmI2/H2O/Et3N) is known to be a particularly powerful reductant, but until now the limiting reducing power has not been determined. A series of unsaturated hydrocarbons with varying half-wave reduction potentials (E(1/2) = -1.6 to -3.4 V, vs SCE) have been treated with SmI2/H2O/Et3N and YbI2/H2O/Et3N, respectively. All hydrocarbons with potentials of -2.8 V or more positive were readily reduced with SmI2/H2O/Et3N, whereas all hydrocarbons with potentials of -2.3 V or more positive were readily reduced using YbI2/H2O/Et3N. This defines limiting values of the chemical reducing power of SmI2/H2O/Et3N to -2.8 V and of YbI2/H2O/Et3N to -2.3 V vs SCE. PMID- 16468809 TI - Scope and facial selectivity of the Prins-pinacol synthesis of attached rings. AB - Using a B-alkyl Suzuki cross-coupling as a key step, a concise and stereocontrolled synthesis of five- to eight-membered triisopropylsiloxy ethers having (2Z)-(6,6-dimethoxyhexylidene) or (2Z)-(5,5-dimethoxypentylidene) side chains was developed. Prins-pinacol reactions of these precursors promoted by SnCl4 provide bicyclic products in which 5-, 6-, or 7-membered rings are joined by a C-C single bond. Synthetically challenging attached ring systems in which both rings are chiral can be prepared in this fashion with high stereo- and enantioselectivity. Stabilizing through-space electrostatic interactions between the alpha-alkoxycarbenium ion and an axially positioned oxygen substituent are believed to play a significant role in organizing the transition structure of the Prins cyclization. PMID- 16468810 TI - Highly diastereoselective enolate addition of O-protected alpha-hydroxyacetate to (S(R))-tert-butanesulfinylimines: synthesis of taxol side chain. AB - The taxol side chain (S(R),2R,3S)-N-tert-butanesulfinyl-O-Boc-3-phenylisoserine benzyl ester 4c was synthesized through a lithium enolate addition of O-Boc-alpha hydroxyacetate benzyl ester 5c to benzylidene (S(R))-tert-butanesulfinamide 6a in excellent yield and diastereoselectivity. By similar approach, a series of enantiopure 3-substituted isoserine benzyl esters 4 useful for the semi-syntheses of taxol derivatives were also prepared in high to excellent yields and diastereoselectivities. The diastereoselective addition mechanism was discussed on the basis of the experimental observation. PMID- 16468812 TI - A conformationally flexible, urea-based tripodal anion receptor: solid-state, solution, and theoretical studies. AB - Tripodal tris(urea) cationic receptors 1 and 2 containing p-tolyl or octyl substituents, respectively, have been synthesized, and their association behavior with anionic guests has been studied via a variety of methods. The receptors are based around a hexasubstituted aryl core and contain both urea and pyridinium functionalities. For 1:1 complexes, anions reside within the central cavity of the host species, held by hydrogen bonds from both NH and CH donors. The following host-anion complexes have been characterized by X-ray crystallography: 1-(Br)3, 1-(PF6)3.2(CH3)2CO, and 1-(NO3)1.5(PF6)1.5. Each structure contains the receptor in a significantly different geometry, highlighting the anion-dependent conformational flexibility of 1. Solution 1H NMR spectroscopic titrations have shown the two host species to display significant affinity for both halides and hydrogen sulfate and strongly suggest the persistence of CH...X- interactions despite the presence of "stronger" NH donor groups. Variable-temperature 1H NMR studies on the more soluble octyl derivative 2 show that there is a distinct change in conformation associated with the formation of a 1:1 host/guest complex. Computations using density functional theory (with the B3LYP functional) have been employed to aid in understanding the geometry of the 1:1 host/chloride complexes of 1 and 2. These experiments suggest that the lowest energy conformation for 1-Cl is one in which the ureidopyridinium arms are orientated upward forming a cavity that is sealed by CH...pi interactions, effectively forming a unimolecular capsule, whereas for 2 a less symmetrical "2-up, 1-down" geometry is favored. PMID- 16468811 TI - Protonated 2-methyl-1,2-epoxypropane: a challenging problem for density functional theory. AB - Protonated epoxides feature prominently in organic chemistry as reactive intermediates. Herein, we describe 10 protonated epoxides using B3LYP, MP2, and CCSD/6-311++G calculations. Relative to CCSD, B3LYP consistently overestimates the C2-O bond length. Protonated 2-methyl-1,2-epoxypropane is the most problematic species studied, where B3LYP overestimates the C2-O bond length by 0.191 angstroms. Seventeen other density functional methods were applied to this protonated epoxide; on average, they overestimated the CCSD bond length by 0.2 angstroms. We present a range of data that suggest the difficulty for DFT methods in modeling the structure of the titled protonated epoxide lies in the extremely weak C2-O bond, which is reflected in the highly asymmetric charge distribution between the two ring carbons. Protonated epoxides featuring more symmetrical charge distribution and cyclic homologues featuring less ring strain are treated with greater accuracy by B3LYP. Finally, MP2 performed very well against CCSD, deviating in the C2-O bond length at most by 0.009 angstroms; it is, therefore, recommended when computational resources prove insufficient for coupled cluster methods. PMID- 16468813 TI - Reactions of cyclohexenyl iodonium tetrafluoroborate with bromide ion: retardation due to the formation of lambda3-bromoiodane. AB - The reaction of 4-tert-butylcyclohex-1-enyl(phenyl)iodonium tetrafluoroborate (1a) and the 4-chlorophenyl derivative (1b) with bromide ion was examined in methanol, acetonitrile, and chloroform. Products include those derived from the intermediate cyclohexenyl cation as well as 1-bromocyclohexene. Kinetic measurements show that the reaction of 1 is strongly retarded by the added bromide. The curved dependence of the observed rate constant on the bromide concentration is typical of a pre-equilibrium formation of the intermediate adduct with a fast bromide-independent reaction (solvolysis of the iodonium ion). The formation of the adduct, lambda3-bromoiodane, was also confirmed by the UV spectral change. The relative reactivity of the iodonium ion and lambda3 bromoiodane is evaluated to be on the order of 10(2). The bromide substitution product forms both via the S(N)1 reaction of the free iodonium ion and via the ligand coupling of the iodane. PMID- 16468814 TI - One-pot synthesis of cyclic nitrones and their conversion to pyrrolizidines: 7a epi-crotanecine inhibits alpha-mannosidases. AB - [reaction: see text] A new straightforward and inexpensive one-pot procedure is described for the preparation of enantiopure five-membered cyclic nitrones starting from the corresponding lactols. Its efficiency relies on the condensation of unprotected hydroxylamine with readily available lactols and on the chemoselectivity of the subsequent esterification with methanesulfonyl chloride. The targeted enantiomerically pure pyrroline N-oxides are versatile synthetic intermediates: one of the nitrones so-obtained has been converted into new polyhydroxypyrrolizidines, analogues of the alkaloids rosmarinecine and crotanecine, which were assayed for their inhibitory activities toward 22 commercially available glycosidase enzymes. One of them ((-)-7a-epi-crotanecine) is a potent and selective inhibitor of alpha-mannosidases from jack beans and almonds. PMID- 16468815 TI - Synthesis of xyloketal A, B, C, D, and G analogues. AB - A series of demethyl analogues of the natural products xyloketal A, B, C, D, and G have been prepared in a notably direct manner from 3-hydroxymethyl-2-methyl-4,5 dihydrofuran and a series of corresponding phenols. These syntheses featured a boron trifluoride diethyl etherate-promoted electrophilic aromatic substitution reaction as a key step. In the case of the synthesis of analogues of xyloketal A, the process was found to be highly efficient (up to 93% yield). The optimized isolated yield of these reaction products is remarkable in view of the fact that this transformation involves, minimally, six individual reactions. Moreover, these synthetic studies provide significant insight into the possible biogenic origin of the xyloketal natural products. PMID- 16468816 TI - Diversity-oriented synthesis of 3-iodochromones and heteroatom analogues via ICl induced cyclization. AB - The ICl-induced cyclization of heteroatom-substituted alkynones provides a simple, highly efficient approach to various 3-iodochromones and analogues. This process is run under mild conditions, tolerates various functional groups, and generally provides chromones in good to excellent yields. Subsequent palladium catalyzed transformations afford a rapid increase in molecular complexity and a convenient preparation of a wide range of functionally substituted chromones, furans, and polycyclic compounds. Iodothiochromenones and iodoquinolinones are also prepared by similar ICl-induced cyclizations. PMID- 16468817 TI - Synthesis and reactions of 2,2-[60]fullerenoalkanoyl chlorides. AB - 2,2-[60]Fullerenoalkanoyl chlorides (1a-d) were easily and securely prepared from the corresponding 2,2-[60]fullerenoalkanoic acids (2a-d) by the reaction with thionyl chloride in an unusual mixed solvent, CH2Cl2/dioxane. The characterization of 1a-d by 1H and 13C NMR, FT-IR, and MALDI-TOF-MASS was conducted for the first time. The 2,2-[60]fullerenoalkanoyl chlorides thus obtained were readily converted to the corresponding amides and esters in moderate to excellent yields by the condensation with amines and alcohols, respectively. Upon applying the condensation, [60]fullerene-biomolecule hybrids were easily prepared. PMID- 16468819 TI - Highly stereoselective beta-anomeric glycosidation of a 2'-deoxy syn-5' configured 4'-spironucleoside. AB - A direct enantioselective pathway that delivers exclusively the beta-anomer of a 4'-spironucleoside has been successfully developed. The key starting material is the enantiomerically pure dihydroxy lactone 19, which has proven amenable to conversion to glycal 22 via the chloro acetonide. This intermediate is then capped as the 3,5-O-(tetraisopropyldisiloxane-1,3-diyl) glycal. The latter can enter into N-iodosuccinimide-promoted glycosidation with persilylated thymine. Only the beta anomer is formed. Ensuing deiodination and desilylation proceed quantitatively to furnish the targeted, previously elusive anomer. PMID- 16468820 TI - Regio- and stereoselective synthesis of aziridino epoxides from cyclic dienes. AB - Two different routes for the regio- and stereoselective synthesis of aziridino epoxides from cyclic dienes have been explored. The first strategy involves regiospecific aziridination of cyclic diene derivatives and subsequent epoxidation with m-CPBA to yield cis-aziridino epoxides as major products. The second strategy utilizes regiospecific epoxidation of cyclic diene derivatives followed by Sharpless aziridination to provide exclusively trans-aziridino epoxides. Synthesis of both enantiomers of cis-aziridino epoxides from (R)-(-)- and (S)-(+)-carvones are also reported. PMID- 16468821 TI - Synthesis and reactivity of laquinimod, a quinoline-3-carboxamide: intramolecular transfer of the enol proton to a nitrogen atom as a plausible mechanism for ketene formation. AB - 5-Chloro-N-ethyl-1,2-dihydro-4-hydroxy-1-methyl-2-oxo-N-phenyl-3 quinolinecarboxamide (laquinimod, 2) is an oral drug in clinical trials for the treatment of multiple sclerosis. The final step in the synthesis of 2 is a high yielding aminolysis reaction of ester 1 with N-ethylaniline. An equilibrium exists between 1 and 2, and removal of formed methanol during the reaction is a prerequisite for obtaining high yields of 2 from 1. The reactivity of 1 and 2 is explained by a mechanistic model that involves a transfer of the enol proton to the exocyclic carbonyl substituent with concomitant formation of ketene 3. This proton transfer is especially facilitated for 2 because the intramolecular hydrogen bond to the carbonyl oxygen is weakened due to steric interactions. Both 1 and 2 undergo solvolosis reactions that obey first-order reaction kinetics, further supporting the theory that these two molecules are able to decompose unimolecularly into ketene 3. The solvent-dependent spectroscopic features of 2 indicate that the molecule mainly resides in two conformations. One conformation is favored in nonpolar solvents and is likely the result of intramolecular hydrogen bonding. The other conformation is favored in polar solvents and probably exhibits less intramolecular hydrogen bonding. PMID- 16468818 TI - Synthesis and characterization of a [3-15N]-labeled cis-syn thymine dimer containing DNA duplex. AB - Cis-syn thymine dimers are the major photoproducts of DNA and are the principal cause of mutations induced by sunlight. To better understand the nature of base pairing with cis-syn thymine dimers, we have synthesized a decamer oligodeoxynucleotide (ODN) containing a cis-syn thymine dimer labeled at the N3 of both T's with 15N by two efficient routes from [3-15N]-thymidine phosphoramidite. In the postsynthetic irradiation route, an ODN containing an adjacent pair of [3-15N]-labeled T's was irradiated and the cis-syn dimer containing ODN isolated by HPLC. In the mixed building block route, a mixture of cis-syn and trans-syn dimer-containing ODNs was synthesized from a mixture of [3 15N]-labeled thymine dimer phosphoramidites after which the cis-syn dimer containing ODN was isolated by HPLC. The N3-nitrogen and imino proton signals of an (15)N-labeled thymine dimer-containing decamer duplex were assigned by 2D 1H 15N heterocorrelated HSQC NMR spectroscopy, and the 15N-1H coupling constant was found to be 1.8 Hz greater for the 5'-T than for the 3'-T. The larger coupling constant is indicative of weaker H-bonding that is consistent with the more distorted nature of the 5'-base pair found in solution state NMR and crystallographic structures. PMID- 16468822 TI - On the mechanism of the Skraup-Doebner-Von Miller quinoline synthesis. AB - The mechanism of the formation of substituted quinolines from anilines and alpha,beta-unsaturated ketones has been studied by the use of 13C-labeled ketones in cross-over experiments. In the reaction of doubly labeled 13C(2,4) mesityl oxide, a 100% scrambling of the label in the quinoline product was observed, whereas only a small (5-10%) amount of the starting mesityl oxide showed scrambling of the label. Similarly, the reaction of triply labeled pulegone clearly shows that the label in the product is 100% scrambled, whereas the label in the starting pulegone is retained. On the basis of these studies, a mechanistic pathway for the Skraup quinoline synthesis is proposed that involves a fragmentation-recombination mechanism. The aniline component condenses with the alpha,beta-unsaturated ketone initially in a conjugate fashion, followed by a fragmentation to the corresponding imine and the ketone itself. These fragments recombine to form the quinoline product. PMID- 16468823 TI - Solvent and base dependence of copper-free palladium-catalyzed cross-couplings between terminal alkynes and arylic iodides: development of efficient conditions for the construction of gold(III)/free-base porphyrin dimers. AB - In this paper, our attempts to optimize the Heck alkynylation (copper-free Sonogashira) reaction are presented. An efficient copper-free coupling protocol was needed for the synthesis of gold/zinc porphyrin dimers because previous methods had failed. Previous studies have usually focused on ligands, whereas this work focuses on the choice of solvent and base. The catalytic system throughout the investigation was formed from the stable precursor [Pd2(dba)3.CHCl3] together with the ligand triphenylarsine, an easy-to-handle, air-stable ligand. A model study was conducted to examine the dependence of the Heck alkynylation on the solvent and base. The most successful modification proved to be the addition of methanol, as a cosolvent, in combination with a nucleophilic tertiary base. The success of the methanol additive is hypothesized to be caused by the presence of a rate-determining deprotonation step featuring a charge-separated transition state. Finally, the very high yielding and successful synthesis of a series of porphyrin systems using these new conditions is presented. For the first time, gold porphyrin substrates could efficiently be coupled in Heck alkynylation reactions. PMID- 16468824 TI - Kinetic acidity of supramolecular imidazolium salts-effects of substituent, preorientation, and counterions on H/D exchange rates. AB - The deprotonation of imidazolium salts to N-heterocyclic carbenes is often a decisive step in modern catalytic reactions. Therefore, we studied the H/D exchange of the C2 H of 15 imidazolium-substituted calix[4]arenes and 11 nonmacrocyclic model compounds in methanol/water (97:3). The influence of the counterion, substitution directly on the imidazolium unit or on the preorientating calixarene backbone could be studied. The observed exchange rates might give a rational for the suitability of the imidazolium salts as precursors in the Suzuki coupling. PMID- 16468825 TI - S(N)2 reaction of sulfur nucleophiles with hindered sulfamidates: enantioselective synthesis of alpha-methylisocysteine. AB - The work described here demonstrates that the five-membered cyclic alpha methylisoserine-derived sulfamidate, (R)-1, behaves as an excellent chiral building block for the ring-opening reaction by S(N)2 attack with sulfur nucleophiles at the quaternary carbon. As a synthetic application of this methodology, and to show that this sulfamidate is a valuable starting material, the synthesis of two new alpha-methylisocysteine derivatives has been carried out to cover the lack of alpha- and beta-methylated amino acids that incorporate the cysteine or isocysteine skeleton. These compounds are two new alpha,alpha disubstituted beta-amino acids (beta(2,2)-amino acids), and the synthetic routes involve nucleophilic ring opening followed by acid hydrolysis. PMID- 16468826 TI - Stereoselective uncatalyzed synthesis of 2,3-unsaturated-4-n-substituted-beta-O glycosides by means of a new D-galactal-derived N-(mesyl)-aziridine. AB - The reaction of the new D-galactal-derived allylic aziridine 1beta with O nucleophiles (alcohols and monosaccharides) affords, in a high to complete beta stereoselectivity, the corresponding 2,3-unsaturated-beta-O-glycosides bearing a beta-N-functionality at C4. PMID- 16468827 TI - Role of electron density and magnetic couplings on the nucleus-independent chemical shift (NICS) profiles of [2.2]paracyclophane and related species. AB - The nucleus-independent chemical shifts (NICS) and electron density profiles along the inner and outer regions defined by the two stacked aromatic rings of [2.2]paracyclophane have been analyzed and compared to those of free benzene and p-xylene and benzene dimers taken as reference models. It is found that stacked aromatic rings show a reduction of the NICS indicator of aromaticity as compared to the same free aromatic systems. This decrease of the NICS values upon stacking is not due to an increase of the electron density in the inner region between the rings (as claimed in a previous work) but is related to the magnetic couplings between superimposed rings that affect this measure of local aromaticity. The increase of local aromaticity in superimposed aromatic rings indicated by NICS is not real but the result of the coupling between the magnetic fields generated by the two stacked rings. This result warns about the use of NICS as a descriptor of aromaticity for species having superimposed aromatic rings. PMID- 16468829 TI - Symmetrical bisquinolones via metal-catalyzed cross-coupling and homocoupling reactions. AB - Functionalized 4,4'-bisquinolones can be efficiently synthesized by microwave assisted palladium(0)-catalyzed one-pot borylation/Suzuki cross-coupling reactions or via nickel(0)-mediated homocouplings of 4-chloroquinolin-2(1H)-one precursors. Both methods are also applicable to other types of symmetrical biaryls. PMID- 16468828 TI - Efficient and green telescoped process to 2-methoxy-3-methyl-[1,4]benzoquinone. AB - A telescoped process for the preparation of 2-methoxy-3-methyl-[1,4]benzoquinone is disclosed. When this novel process is compared to the prevailing method that utilizes Na2Cr2O7 as the oxidant, the novel process represents a high yielding (95%), green, and environmentally benign alternative with H2O2 and HNO3 as the oxidants and CH3COOH as the reaction medium. PMID- 16468830 TI - Heck, direct arylation, and hydrogenation: two or three sequential reactions from a single catalyst. AB - Palladium-catalyzed tandem multifunctional reactions leading to the synthesis of substituted biaryl molecules have been developed including tandem Heck-direct arylation and tandem-sequential Heck-direct arylation-hydrogenation. These reactions occur in good yield and have been employed in the synthesis of a cytotoxic biaryl compound. PMID- 16468831 TI - Effective asymmetric epoxidation of styrenes by chiral dioxirane. AB - High enantioselectivity (80-92% enantiomeric excess (ee)) has been obtained for the epoxidation of various styrenes using an easily prepared ketone (4) catalyst. PMID- 16468832 TI - First synthesis of free cholesterol-BODIPY conjugates. AB - Analogues of cholesterol (compounds 1 and 2) and coprostanol (compound 3) containing the BODIPY fluorophore in the aliphatic tail of the free sterol have been synthesized starting with bisnorcholenic acid, cholenic acid 3beta-acetate, and lithocholic acid, respectively. An ester linkage joining the fluorophore to the sterol nucleus interfered with the ability of the fluorescent sterol to pack with phospholipids in monolayers. However, an analogue in which the linker was devoid of polar atoms exhibited a substantially similar physical behavior to cholesterol in model membranes with respect to localization in raft domains. PMID- 16468833 TI - A general and chemoselective synthesis of phosphoramidates through reaction of silylated nucleoside di- and triphosphates with silylated amines containing multifunctional groups. AB - An approach to various phosphoramidates has been developed through one-pot chemoselective reaction of nucleoside di- and triphosphates with 3'-amino-3' deoxythymidine, d-glucosamine, and peptides mediated by trimethylsilyl chloride, and the corresponding conjugates via the linkage of phosphate were obtained. The method could be generally applied for the synthesis of phosphoramidates without any preprotection of polyphosphates and amines containing multifunctional groups. PMID- 16468834 TI - An improved synthesis of pyran-3,5-dione: application to the synthesis of ABT 598, a potassium channel opener, via Hantzsch reaction. AB - Ketoester 1 is cyclized to give pyran-3,5-dione 2 in 78% yield using a parallel addition of ketoester 1 and base NaO(t)Bu in refluxing THF. Compared to the previously reported procedures, these optimized conditions have significantly increased the yield of this transformation and the quality of pyran 2 and prove to be suitable for large-scale preparation. An application of 2 to the synthesis of ABT-598, a potassium channel opener, is demonstrated. PMID- 16468835 TI - Direct reaction of dibromoacetic acid with aldehydes promoted by samarium diiodide: an easy, efficient, and rapid synthesis of (E)-alpha,beta-unsaturated carboxylic acids with total stereoselectivity. AB - A promoted SmI2 direct reaction of dibromoacetic acid with different aldehydes, followed by an elimination reaction also promoted by samarium diiodide, affords (E)-alpha,beta-unsaturated carboxylic acids 2 with total stereoselectivity. A mechanism to explain this transformation is proposed. PMID- 16468836 TI - Ligand effects in the synthesis of N-heterocycles by intramolecular Heck reactions. AB - Chemo- and regioselectivity of intramolecular Heck reactions are dependent on the type of ligand employed. Six- to eight-membered benzolactams are produced in good yields using PPh3 as ligand. In contrast, a biaryl coupling occurred preferentially under ligandless conditions to form a dihydrophenanthridine product. Conformations of the seven- and eight-membered benzolactams in the solid state were examined by X-ray crystallography. PMID- 16468837 TI - Synthesis of 2-arylcycloalka-2,4-dienones using sulfone-based methodology. AB - Addition of the anion derived from (phenylthiomethyl)phenyl sulfone to a selection of aryl omega-alkenyl ketones gives adducts that via a sequence of Lewis acid catalyzed rearrangement, alpha-allylation, and metathesis give rise to 2-thio-4-cycloalkenones. These in turn give cycloalkadienones upon oxidation and elimination. An attempt to develop a polymer-supported variant fails because of the reversibility of sulfone anion addition. PMID- 16468838 TI - Synthesis of deuterium-labeled derivatives of dimethylallyl diphosphate. AB - Short practical syntheses for five deuterium-labeled derivatives of dimethylallyl diphosphate (DMAPP) useful for enzymological studies are reported. These include the preparation of the C1-labeled derivatives (R)-[1-2H]3-methylbut-2-enyl diphosphate ((R)-[1-2H]1-OPP) and (S)-[1-2H]3-methylbut-2-enyl diphosphate ((S) [1-2H]1-OPP), the C2-labeled derivative [2-2H]3-methylbut-2-enyl diphosphate ([2 2H]1-OPP), and the methyl-labeled derivatives (E)-[4,4,4-2H3]3-methylbut-2-enyl diphosphate ((E)-[4,4,4-2H3]1-OPP) and (Z)-[4,4,4-2H3]3-methyl-but-2-enyl diphosphate ((Z)-[4,4,4-2H3]1-OPP). PMID- 16468839 TI - Photochemical ring expansion of 4-azidouracil: a route to 5H-1,3,5-triazepin-2,4 dione in the nucleoside series. AB - Under aqueous conditions, 4-azidouracil/tetrazolo[1,5-c]pyrimidin-5(6H)-one nucleosides undergo a very efficient photochemical nitrogen elimination and ring expansion to 1,3,5-triazepin-2,4-dione nucleosides whose structure has been confirmed by X-ray crystallography. In contrast, when the photolysis was attempted under anhydrous conditions in the presence of a nucleophile, a ring contraction reaction occurred, affording 2-oxoimidazolone nucleosides. A mechanism to account for the formation of ring expansion and contraction reactions and involving a carbodiimide intermediate is proposed which is reminiscent of the known photochemical behavior of 2-azidopyridines/tetrazolo[1,5 a]pyridines. PMID- 16468840 TI - Regioselective 6-endo cyclizations of 2-indolylacyl radicals: total synthesis of the pyrido[4,3-b]carbazole alkaloid guatambuine. AB - A regioselective 6-endo reductive cyclization of 2-indolylacyl radicals constitutes the key step of a straightforward synthetic entry to the olivacine skeleton, illustrated by a total synthesis of the tetrahydropyridine alkaloid guatambuine. PMID- 16468841 TI - Synthesis of hydantoins from enantiomerically pure alpha-amino amides without epimerization. AB - A method for the preparation of enantiomerically pure hydantoins from optically pure alpha-amino amides utilizing triphosgene is described. We also propose that the racemization observed with 1,1'-carbonyldiimidazole (CDI) for this type of reaction is due to the imidazole carbamate intermediates. PMID- 16468845 TI - Generalizations of the Hohenberg-Kohn theorem: I. Legendre transform constructions of variational principles for density matrices and electron distribution functions. AB - Given a general, N-particle Hamiltonian operator, analogs of the Hohenberg-Kohn theorem are derived for functions that are more general than the particle density, including density matrices and the diagonal elements thereof. The generalization of Lieb's Legendre transform ansatz to the generalized Hohenberg Kohn functional not only solves the upsilon-representability problem for these entities, but, more importantly, also solves the N-representability problem. Restricting the range of operators explored by the Legendre transform leads to a lower bound on the true functional. If all the operators of interest are incorporated in the restricted maximization, however, the variational principle dictates that exact results are obtained for the systems of interest. This might have important implications for practical work not only for density matrices but also for density functionals. A follow-up paper will present a useful alternative approach to the upsilon- and N-representability problems based on the constrained search formalism. PMID- 16468846 TI - Using simultaneous diagonalization and trace minimization to make an efficient and simple multidimensional basis for solving the vibrational Schrodinger equation. AB - In this paper we improve the product simultaneous diagonalization (SD) basis method we previously proposed [J. Chem. Phys. 122, 134101 (2005)] and applied to solve the Schrodinger equation for the motion of nuclei on a potential surface. The improved method is tested using coupled complicated Hamiltonians with as many as 16 coordinates for which we can easily find numerically exact solutions. In a basis of sorted products of one-dimensional (1D) SD functions the Hamiltonian matrix is nearly diagonal. The localization of the 1D SD functions for coordinate qc depends on a parameter we denote alphac. In this paper we present a trace minimization scheme for choosing alphac to nearly block diagonalize the Hamiltonian matrix. Near-block diagonality makes it possible to truncate the matrix without degrading the accuracy of the lowest energy levels. We show that in the sorted product SD basis perturbation theory works extremely well. The trace minimization scheme is general and easy to implement. PMID- 16468847 TI - Formal relations connecting different approaches to calculate relativistic effects on molecular magnetic properties. AB - In the present work a set of formal relations connecting different approaches to calculate relativistic effects on magnetic molecular properties are proven. The linear response (LR) within the elimination of the small component (ESC), Breit Pauli, and minimal-coupling approaches are compared. To this end, the leading order ESC reduction of operators within the minimal-coupling four-component approach is carried out. The equivalence of all three approaches within the ESC approximation is proven. It is numerically verified for the NMR nuclear-magnetic shielding tensor taking HX and CH3X (X=Br,I) as model compounds. Formal relations proving the gauge origin invariance of the full relativistic effect on the NMR nuclear-magnetic shielding tensor within the LR-ESC approach are presented. PMID- 16468848 TI - Oscillator strengths of helium computed using Monte Carlo methods. AB - We have optimized trial wave functions for the three lowest states of the helium atom with symmetry 1S, 1P, 1D, 3S, 3P, and 3D using variational Monte Carlo methods. With these wave functions we then computed dipole oscillator strengths for the 1S-1P, 1P-1D, 3S-3P, and 3P-3D transitions using the length, velocity, and acceleration forms. Our values are in good agreement with the best results found in the literature. PMID- 16468849 TI - Interpolating moving least-squares methods for fitting potential-energy surfaces: further improvement of efficiency via cutoff strategies. AB - In standard applications of interpolating moving least squares (IMLS) for fitting a potential-energy surface (PES), all available ab initio points are used. Because remote ab initio points negligibly influence IMLS accuracy and increase IMLS time-to-solution, we present two methods to locally restrict the number of points included in a particular fit. The fixed radius cutoff (FRC) method includes ab initio points within a hypersphere of fixed radius. The density adaptive cutoff (DAC) method includes points within a hypersphere of variable radius depending on the point density. We test these methods by fitting a six dimensional analytical PES for hydrogen peroxide. Both methods reduce the IMLS time-to-solution by about an order of magnitude relative to that when no cutoff method is used. The DAC method is more robust and efficient than the FRC method. PMID- 16468850 TI - Gradient symplectic algorithms for solving the radial Schrodinger equation. AB - The radial Schrodinger equation for a spherically symmetric potential can be regarded as a one-dimensional classical harmonic oscillator with a time-dependent spring constant. For solving classical dynamics problems, symplectic integrators are well known for their excellent conservation properties. The class of gradient symplectic algorithms is particularly suited for solving harmonic-oscillator dynamics. By use of Suzuki's rule [Proc. Jpn. Acad., Ser. B: Phys. Biol. Sci. 69, 161 (1993)] for decomposing time-ordered operators, these algorithms can be easily applied to the Schrodinger equation. We demonstrate the power of this class of gradient algorithms by solving the spectrum of highly singular radial potentials using Killingbeck's method [J. Phys. A 18, 245 (1985)] of backward Newton-Ralphson iterations. PMID- 16468851 TI - Sources of error in electronic structure calculations on small chemical systems. AB - The sources of error in electronic structure calculations arising from the truncation of the one-particle and n-particle expansions are examined with very large correlation consistent basis sets, in some cases up through valence 10-zeta quality, and coupled cluster methods, up through connected quadruple excitations. A limited number of full configuration interaction corrections are also considered. For cases where full configuration interaction calculations were unavailable or prohibitively expensive, a continued fraction approximation was used. In addition, errors arising from corevalence and relativistic corrections are also probed for a number of small chemical systems. The accuracies of several formulas for estimating total energies and atomization energies in the complete basis set limit are compared in light of the present large basis set findings. In agreement with previous work, the CCSD(T) method is found to provide results that are closer to the CCSDTQ and full configuration-interaction results than the less approximate CCSDT method. PMID- 16468852 TI - Third-order multireference perturbation theory: the n-electron valence state perturbation-theory approach. AB - A formulation of the n-electron valence state perturbation theory (NEVPT) at the third order of perturbation is presented. The present implementation concerns the so-called strongly contracted variant of NEVPT, where only a subspace of the first-order interacting space is taken into account. The resulting strongly contracted NEVPT3 approach is discussed in three test cases: (a) the energy difference between the 3B1 and 1A1 states of the methylene molecule, (b) the potential-energy curve of the N2 molecule ground state, and (c) the chromium dimer (Cr2) ground-state potential-energy profile. Particular attention is devoted to the last case where large basis sets comprising also h orbitals are adopted and where remarkable differences between the second- and third-order results show up. PMID- 16468853 TI - Quadratic string method for determining the minimum-energy path based on multiobjective optimization. AB - Based on a multiobjective optimization framework, we develop a new quadratic string method for finding the minimum-energy path. In the method, each point on the minimum-energy path is minimized by integration in the descent direction perpendicular to path. Each local integration is done on a quadratic surface approximated by a damped Broyden-Fletcher-Goldfarb-Shanno updated Hessian, allowing the algorithm to take many steps between energy and gradient calls. The integration is performed with an adaptive step-size solver, which is restricted in length to the trust radius of the approximate Hessian. The full algorithm is shown to be capable of practical superlinear convergence, in contrast to the linear convergence of other methods. The method also eliminates the need for predetermining such parameters as step size and spring constants, and is applicable to reactions with multiple barriers. The effectiveness of this method is demonstrated for the Muller-Brown potential, a seven-atom Lennard-Jones cluster, and the enolation of acetaldehyde to vinyl alcohol. PMID- 16468854 TI - Inner-shell ionizations and satellites studied by the open-shell reference symmetry-adapted cluster/symmetry-adapted cluster configuration-interaction method. AB - Open-shell reference version of the symmetry-adapted cluster (SAC) and SAC configuration-interaction (CI) methods, termed open-shell reference (OR)-SAC and OR-SAC-CI methods, are developed and applied to inner-shell ionizations of CH4, NH3, H2O, and HF. The inner-shell ionization potentials and spectra calculated by the OR-SAC and OR-SAC-CI methods are in excellent agreement with the experimental data. Including both of the electron correlation and orbital relaxation is important for quantitative agreements. Timing comparisons with the SAC-CI general R calculations that give similar high accuracies show an efficiency of the present OR-SAC and OR-SAC-CI methods. PMID- 16468855 TI - Diffusion and velocity relaxation of a Brownian particle immersed in a viscous compressible fluid confined between two parallel plane walls. AB - The diffusion tensor and velocity correlation function of a Brownian particle immersed in a viscous compressible fluid confined between two parallel plane walls are calculated in point approximation. The fluid is assumed to satisfy stick boundary conditions at the walls. It is found that the velocity correlation function decays asymptotically with a negative t(-2) long-time tail due to coupling to overdamped sound waves. The coefficient of the long-time tail is calculated and shown to be independent of fluid viscosity. PMID- 16468856 TI - Efficient computation of the first passage time distribution of the generalized master equation by steady-state relaxation. AB - The generalized master equation or the equivalent continuous time random walk equations can be used to compute the macroscopic first passage time distribution (FPTD) of a complex stochastic system from short-term microscopic simulation data. The computation of the mean first passage time and additional low-order FPTD moments can be simplified by directly relating the FPTD moment generating function to the moments of the local FPTD matrix. This relationship can be physically interpreted in terms of steady-state relaxation, an extension of steady-state flow. Moreover, it is amenable to a statistical error analysis that can be used to significantly increase computational efficiency. The efficiency improvement can be extended to the FPTD itself by modelling it using a gamma distribution or rational function approximation to its Laplace transform. PMID- 16468857 TI - Local stress and heat flux in atomistic systems involving three-body forces. AB - Local densities of fundamental physical quantities, including stress and heat flux fields, are formulated for atomistic systems involving three-body forces. The obtained formulas are calculable within an atomistic simulation, in consistent with the conservation equations of thermodynamics of continuum, and can be applied to systems with general two- and three-body interaction forces. It is hoped that this work may correct some misuse of inappropriate formulas of stress and heat flux in the literature, may clarify the definition of site energy of many-body potentials, and may serve as an analytical link between an atomistic model and a continuum theory. Physical meanings of the obtained formulas, their relation with virial theorem and heat theorem, and the applicability are discussed. PMID- 16468858 TI - Explicitly correlated second-order perturbation theory using density fitting and local approximations. AB - Three major obstacles in electronic structure theory are the steep scalings of computer time with respect to system size and basis size and the slow convergence of correlation energies in orbital basis sets. Three solutions to these are, respectively, local methods, density fitting, and explicit correlation; in this work, we combine all three to produce a low-order scaling method that can achieve accurate MP2 energies for large systems. The errors introduced by the local approximations into the R12 treatment are analyzed for 16 chemical reactions involving 21 molecules. Weak pair approximations, as well as local resolution of the identity approximations, are tested for molecules with up to 49 atoms, over 100 correlated electrons, and over 1000 basis functions. PMID- 16468859 TI - A simple natural orbital mechanism of "pure" van der Waals interaction in the lowest excited triplet state of the hydrogen molecule. AB - A treatment of van der Waals (vdW) interaction by density-matrix functional theory requires a description of this interaction in terms of natural orbitals (NOs) and their occupation numbers. From an analysis of the configuration interaction (CI) wave function of the 3Sigmau + state of H2 and the exact NO expansion of the two-electron triplet wave function, we demonstrate that the construction of such a functional is straightforward in this case. A quantitative description of the vdW interaction is already obtained with, in addition to the standard part arising from the Hartree-Fock determinant /1sigmag(r1)1sigmau(r2)/, only two additional terms in the two-electron density, one from the first "excited" determinant /2sigmag(r1)2sigmau(r2)/ and one from the state of 3Sigmau + symmetry belonging to the (1pig)1(1piu)1 configuration. The potential-energy curve of the 3Sigmau + state calculated around the vdW minimum with the exact density-matrix functional employing only these eight NOs and NO occupations is in excellent agreement with the full CI one and reproduces well the benchmark potential curve of Kolos and Wolniewicz [J. Chem. Phys. 43, 2429 (1965)]. The corresponding terms in the two-electron density rho2(r1,r2), containing specific products of NOs combined with prefactors that depend on the occupation numbers, can be shown to produce exchange-correlation holes that correspond precisely to the well-known intuitive picture of the dispersion interaction as an instantaneous dipole-induced dipole (higher multipole) effect. Indeed, (induced) higher multipoles account for almost 50% of the total vdW bond energy. These results serve as a basis for both a density-matrix functional theory of van der Waals bonding and for the construction of orbital-dependent functionals in density-functional theory that could be used for this type of bonding. PMID- 16468860 TI - A general framework for non-Boltzmann Monte Carlo sampling. AB - Non-Boltzmann sampling (NBS) methods have been extensively employed in recent years, mainly due to their ability to enhance ergodicity in simulations of complex systems. In addition, they make possible reliable computation of equilibrium properties (ensemble averages, free-energy differences, and potentials of mean force) over continuous ranges of thermodynamic conditions. In this work, we put forward a general and systematic framework for NBS methods that allows a single set of equations and procedures to be applied to diverse systems. Moreover, we show how to exploit simulation data most effectively by obtaining continuous profiles of any mechanical properties, including structural quantities not directly related to the ensemble parameters. Finally, we demonstrate the usefulness of the developed formulation by applying it to spin systems, Lennard Jones fluids, and a model protein molecule (both in isolation and in the proximity of a flat wall). PMID- 16468861 TI - Ordering of limits in the Jarzynski equality. AB - We consider the sampling problems encountered in computing free-energy differences using Jarzynski's nonequilibrium work relation [Phys. Rev. Lett. 56, 2690 (1997)]. This relation expresses the free-energy change of a system, on which finite-time work is done, as an average over all possible trajectories of the system. This average can then be expressed as a cumulant expansion of the work. We study the scaling of these cumulants with an appropriately defined measure of phase-space accessibility epsilon and particle number N for two simple changes in state. We find that the cumulant expansion is slowly convergent for predominantly entropic processes, those where epsilon is considerably altered during the course of the process. An accurate determination of the free-energy change requires some knowledge of the behavior of the tails of the work distribution associated with the process. Jarzynski's irreversible work relation is only valid with the correct ordering of the infinite limits of N and epsilon, clarifying the regime of its applicability. PMID- 16468862 TI - Validation of Markov state models using Shannon's entropy. AB - Markov state models are kinetic models built from the dynamics of molecular simulation trajectories by grouping similar configurations into states and examining the transition probabilities between states. Here we present a procedure for validating the underlying Markov assumption in Markov state models based on information theory using Shannon's entropy. This entropy method is applied to a simple system and is compared with the previous eigenvalue method. The entropy method also provides a way to identify states that are least Markovian, which can then be divided into finer states to improve the model. PMID- 16468863 TI - Generation and characterization of highly vibrationally excited molecular beam. AB - A simple method to generate and characterize a pure highly vibrationally excited azulene molecular beam is demonstrated. Azulene molecules initially excited to the S4 state by 266-nm UV photons reach high vibrationally excited levels of the ground electronic state upon rapid internal conversion from the S4 electronically excited state. VUV laser beams at 157 and 118 nm, respectively, are used to characterize the relative concentrations of the highly vibrationally excited azulene and the rotationally and vibrationally cooled azulene in the molecular beam. With a laser intensity of 34 mJ/cm2, 75% of azulene molecules absorb a single 266-nm photon and become highly vibrationally excited molecules. The remaining ground-state azulene molecules absorb two or more UV photons, ending up either as molecular cations, which are repelled out of the beam by an electric field, or as dissociation fragments, which veer off the molecular-beam axis. No azulene without absorption of UV photons is left in the molecular beam. The molecular beam that contains only highly vibrationally excited molecules and carrier gas is useful in various experiments related to the studies of highly vibrationally excited molecules. PMID- 16468864 TI - Energy transfer of highly vibrationally excited azulene: collisions between azulene and krypton. AB - The energy-transfer dynamics between highly vibrationally excited azulene molecules and Kr atoms in a series of collision energies (i.e., relative translational energies 170, 410, and 780 cm(-1)) was studied using a crossed-beam apparatus along with time-sliced velocity map ion imaging techniques. "Hot" azulene (4.66 eV internal energy) was formed via the rapid internal conversion of azulene initially excited to the S4 state by 266-nm photons. The shapes of the collisional energy-transfer probability density functions were measured directly from the scattering results of highly vibrationally excited or hot azulene. At low enough collision energies an azulene-Kr complex was observed, resulting from small amounts of translational to vibrational-rotational (T-VR) energy transfer. T-VR energy transfer was found to be quite efficient. In some instances, nearly all of the translational energy is transferred to vibrational-rotational energy. On the other hand, only a small fraction of vibrational energy is converted to translational energy (V-T). The shapes of V-T energy-transfer probability density functions were best fit by multiexponential functions. We find that substantial amounts of energy are transferred in the backward scattering direction due to supercollisions at high collision energies. The probability for supercollisions, defined arbitrarily as the scattered azulene in the region 160 degrees 2000 cm(-1) is 1% and 0.3% of all other collisions at collision energies 410 and 780 cm(-1), respectively. PMID- 16468865 TI - A quasiclassical trajectory study for the N(4S)+O2(X 3Sigmag-)-->NO(X 2Pi)+O(3P) reaction on the new 2A' and 4A' potential-energy surfaces. AB - A quasiclassical trajectory study with the sixth-order explicit symplectic algorithm of the N(4S)+O2(X 3Sigmag-)-->NO(X 2Pi)+O(3P) atmospheric reaction has been performed by employing the new 2A' and 4A' potential-energy surfaces reported by Sayos et al. [J. Chem. Phys. 117, 670 (2002)]. For the translational temperature considered up to 10,000 K, the larger relative translational energy and the higher rovibrational levels of O2 molecule with respect to the previous works have been taken into account, and a clearer database about the character of the total reaction cross section has been presented in this work. The dependence of microscopic rate constants on the vibrational level of O2 molecule at T=3000, 5000, and 10,000 K has been exhibited, and we can see that the values of log10 k(T,v,J) vary almost linearly with the vibrational level of O2 molecule. The thermal rate constants at the translational temperature between 300 and 10,000 K have been evaluated and compared with the experimental and previous theoretical results. It is found that the thermal rate constants determined in this work have a better agreement with the experimental data and can provide a more valid theoretical reference at the translational temperature considered for the title reaction. PMID- 16468866 TI - Structures and relative stability of medium-sized silicon clusters. IV. Motif based low-lying clusters Si21-Si30. AB - Structures and relative stability of four families of low-lying silicon clusters in the size range of Sin(n=21-30) are studied, wherein two families of the clusters show prolate structures while the third one shows near-spherical structures. The prolate clusters in the first family can be assembled by connecting two small-sized magic clusters Sin (n=6, 7, 9, or 10) via a fused puckered-hexagonal-ring Si9 unit (a fragment of bulk diamond silicon), while those in the second family can be constructed on the basis of a structural motif consisting of a puckered-hexagonal-ring Si6 unit (also a fragment of bulk diamond silicon) and a small-sized magic cluster Sin (n=6, 7, 9, or 10). For Si21-Si29, the predicted lowest-energy clusters (except Si27) exhibit prolate structures. For clusters larger than Si25, the third family of near-spherical clusters becomes energetically competitive. These near-spherical clusters all exhibit endohedral cagedlike structures, and the cages are mostly homologue to the carbon fullerene cages which consist of pentagons and hexagons exclusively. In addition, for Si26-Si30, we construct a new (fourth) family of low-lying clusters which have "Y-shaped" three-arm structures, where each arm is a small-sized magic cluster (Si6, Si7, or Si10). Density-functional calculation with the B3LYP functional shows that this new family of clusters is also energetically competitive, compared to the two prolate and one near-spherical low-lying families. PMID- 16468867 TI - Geometric and electronic structures of metal-substitutional fullerene C59Sm and metal-exohedral fullerenes C60Sm. AB - The geometric and electronic structures of metal-substituted fullerene C59Sm and exohedral fullerenes C60Sm are studied using the density-functional theory. The geometric optimization shows that the replacement of a C atom with a Sm in C60 yields a stable substitutionally doped fullerene C59Sm, and among the five possible optimized geometries for C60Sm, the most favorable exohedral sites are above the center of a hexagon and a pentagon ring. The calculations for electronic structures show that the magnetic moment of Sm is preserved for all the stable structures as tiny hybridization takes place between the orbitals of the Sm atom and those of their neighboring carbons. Because of the small energy gaps and the half occupation of the highest occupied molecular orbitals, all the stable C60Sm isomers are inferred to be conductors. PMID- 16468868 TI - Dissociative ion-pair formation in collisions of fast potassium atoms with benzene and fluorobenzene. AB - Using a crossed molecular-beam technique, electron transfer is studied in collisions of fast potassium atoms with benzene and fluorobenzene molecules. The negative fragment ions formed in the collision region are extracted by a pulsed voltage, and their masses and energy release distributions analyzed by a time-of flight (TOF) method. The dominant fragment from C6H5F is F-. The phenyl ring fragments CH- and C2H- are also observed but with lower intensities. In the case of C6H6 the dominant anionic specie is C2H-. The kinetic-energy release distributions of the various fragments are derived from the width and shapes of the TOF peaks in the spectrum. PMID- 16468869 TI - Coarse-grained interaction potentials for polyaromatic hydrocarbons. AB - Using Kohn-Sham (KS) density-functional theory, we have studied the interaction between various polyaromatic hydrocarbon molecules. The systems range from monocyclic benzene up to hexabenzocoronene (hbc). For several conventional exchange-correlation functionals total potential-energy curves of interaction of the pi-pi stacking hbc dimer are reported. It is found that all pure local density or generalized gradient approximated functionals yield qualitatively incorrect predictions regarding structure and interaction. Inclusion of a nonlocal, atom-centered correction to the KS Hamiltonian enables quantitative predictions. The computed potential-energy surfaces of interaction yield parameters for a coarse-grained potential, which can be employed to study discotic liquid-crystalline mesophases of derived polyaromatic macromolecules. PMID- 16468870 TI - Photoelectron spectroscopy of AlnD2- (n=3-15): observation of chemisorption and physisorption of dideuterium on aluminum cluster anions. AB - Photoelectron spectroscopy is used to investigate aluminum dideuteride cluster anions, AlnD2- (n=3,6-15), produced by laser vaporization of a pure Al target with a D2-seeded helium carrier gas. Comparison between the well-resolved photoelectron spectra of AlnD2- and Aln- reveals the nature of interactions between D2 and Aln-. Depending on the size of the Aln- clusters and their electronic structure, three types of AlnD2- species are observed, dideuteride (dissociative chemisorption), molecular chemisorption, and physisorption. Striking spectral similarities are observed between photoelectron spectra of AlnD2- and Aln- for n=9, 11, 13, and 15, suggesting that D2 is physisorbed on these closed-shell Aln- clusters. For AlnD2- with n=3, 6, 7, and 10, completely different spectra are observed in comparison with the corresponding Aln- clusters, suggesting that the AlnD2- species may be characterized as dideuterides. For AlnD2- with n=8, 12, and 14, in which the Aln- clusters are open shell, the D2 is characterized as chemisorption on the basis of spectral shifts and similarities relative to those of the corresponding Aln- clusters. PMID- 16468871 TI - Methyl iodide clusters observed in gas phase by infrared cavity ring-down spectroscopy: the CH3 bending mode at 8 microm. AB - Infrared spectra of methyl iodide clusters produced in a supersonic jet have been observed in the CH3 bending region at 8 mum by cavity ring-down spectroscopy. The dependence of the spectral features on the mixing ratio of CH3I to He and on the stagnation pressure has allowed us to assign the absorption peaks, with the help of the previous results obtained by matrix-isolation technique [Ito et al., Chem. Phys. Lett. 343, 185 (2001)] and infrared cavity ring-down spectroscopy in the C H stretching region [Ito et al., Chem. Phys. 286, 337 (2003)]. Ab initio calculations at the MP2 level have been carried out up to tetramer to confirm the assignments. It has been found that the frequency shifts upon clustering (relative to monomer) observed in the bending region are not monotonic, in contrast to those in the C-H stretching region. The observed frequency shifts are discussed in terms of dispersion interaction and its variation upon vibrational excitation. PMID- 16468872 TI - Characterization of solvated electrons in hydrogen cyanide clusters: (HCN)n- (n=3, 4). AB - Theoretical studies of the solvated electrons (HCN)n- (n=3, 4) reveal a variety of electron trapping possibilities in the (HCN)n (n=3, 4) clusters. Two isomers for (HCN)3- and four isomers for (HCN)4- are obtained at the MP2aug-cc-pVDZ+dBF (diffusive bond functions) level of theory. In view of vertical electron detachment energies (VDEs) at the CCSD(T) level, the excess electron always "prefers" locating in the center of the system, i.e., the isomer with higher coordination number shows larger VDE value. However, the most stable isomers of the solvated electron state (HCN)3- and (HCN)4- are found to be the linear Cinfinitynu and Dinfinityh structures, respectively, but not the fullyl symmetric structures which have the largest VDE values. PMID- 16468873 TI - Confined helium atom low-lying S states analyzed through correlated Hylleraas wave functions and the Kohn-Sham model. AB - Calculation including the electron correlation effects is reported for the ground 1 1S and lowest triplet 1 3S state energies of the confined helium atom placed at the center of an impenetrable spherical box. While the adopted wave-functional treatment involves optimization of three nonlinear parameters and 10, 20, and 40 linear coefficients contained in wave functions expressed in a generalized Hylleraas basis set that explicitly incorporates the interelectronic distance r12, via a Slater-type exponent and through polynomial terms entering the expansion, the Kohn-Sham model employed here uses the Perdew and Wang exchange correlation functional in its spin-polarized version within the local-density approximation (LDA) with and without the self-interaction correction. All these calculations predict a systematic increase in the singlet-triplet energy splitting toward the high confinement regime, i.e., when the box radius is reduced. By using the variational results as benchmark, it is found that the LDA underestimates the singlet-triplet energy splitting, whereas the self-interaction correction overestimates such a quantity. PMID- 16468875 TI - Spectroscopic and spin-orbit calculations on the SO+ radical cation. AB - Highly correlated ab initio methods were used in order to generate the potential energy curves of the SO+ electronic states correlating to S+(4Su)+O(3Pg) and S+(2Du)+O(3Pg). These curves were used for deducing accurate spectroscopic properties for these electronic states. Our calculations predict the existence of a 2Phi state lying close in energy to the well-characterized b 4Sigma- state and several weakly bound quartet and doublet states located in the 6-9 eV internal energy range not identified yet. The spin-orbit integrals between these electronic states were evaluated using these highly correlated wave functions, allowing the discussion of the metastability and the predissociation processes forming S+ +O in their electronic ground states. Multistep spin-orbit-induced predissociation pathways are suggested. More specifically, the experimentally determined dissociative potential-energy curve [H. Bissantz et al., Z. Phys. D 22, 727 (1992)] proposed to explain the rapid SO+(b 4Sigma-, v> or =13)- >S+(4Su)+O(3Pg) reaction is found to coincide with the 2 4Pi potential-energy curve for short internuclear distances and with the repulsive 1 6Pi state for longer internuclear separations. PMID- 16468874 TI - Low-energy electron scattering from DNA and RNA bases: shape resonances and radiation damage. AB - Calculations are carried out to determine elastic-scattering cross sections and resonance energies for low-energy electron impact on uracil and on each of the DNA bases (thymine, cytosine, adenine, and guanine), for isolated molecules in their equilibrium geometry. Our calculations are compared with the available theory and experiment. We also attempt to correlate this information with experimental dissociation patterns through an analysis of the temporary anion structures that are formed by electron capture in shape resonances. PMID- 16468876 TI - Spectroscopic characterization of the C2-Ne van der Waals complex. AB - Binary complexes of C2 with rare-gas atoms (C2-Rg) have attracted theoretical interest as their potential-energy surfaces are predicted to support linear equilibrium geometries, without the local minimum for the T-shaped geometry that would be expected using a standard pair-potential model. In the present work we have explored the properties of C2-Ne using laser-induced fluorescence detection of the D 1Sigmau +-X 1Sigmag + transition. Bands of the complex were observed in association with the monomer 0-0 and 1-1 transitions. Rotationally resolved data yielded rotational constants of B'=0.099(3) cm(-1) and B"=0.100(3) cm(-1) for the excited and ground states, respectively. Analysis of the rovibrational energy level structure for C2(D)-Ne indicates that the complex has a linear equilibrium structure with a barrier to internal rotation of approximately 15 cm(-1). Data for the ground state validate a recent high-level ab initio calculation of the potential-energy surface for C2(X)-Ne. PMID- 16468877 TI - IR laser manipulation of cis<-->trans isomerization of 2-naphthol and its hydrogen-bonded clusters. AB - The cis<-->trans isomerization reaction has been carried out for 2-naphthol and its hydrogen (H) bonded clusters by infrared (IR) laser in the electronic excited state (S1) in supersonic jets. A specific isomer in the jet was pumped to the X-H stretching vibration in the S1 state, where X refers to C, O, or N atom, by using a stepwise UV-IR excitation, and the dispersed emission spectra of the excited species or generated fragments were observed. It was found that the isomerization occurs only in the H-bonded clusters but a bare molecule does not exhibit the isomerization in the examined energy region of Ev< or =3610 cm(-1), indicating a reduction of the isomerization barrier height upon the H bonding. The relative yield of the isomerization was observed as a function of internal energy. The isomerization yield was found to be very high at the low IR frequency excitation, and was rapidly reduced with the IR frequency due to the competition of the dissociation of the H bond within the isomer. Density-functional theory (DFT) and time-dependent DFT calculations were performed for estimating the barrier height of the isomerization for bare 2-naphthol and its cluster for electronic ground and excited states. The calculation showed that the isomerization barrier height is highly dependent on the electronic states. However, the reduction of the height upon the hydrogen bonding was not suggested at the level of our calculation. PMID- 16468878 TI - CH stretch/internal rotor dynamics in ethyl radical: high-resolution spectroscopy in the CH3-stretch manifold. AB - High-resolution IR absorption spectra of supersonically cooled ethyl radicals (Trot approximately 20 K) have been obtained in a slit supersonic jet discharge expansion, revealing first rotationally resolved data for CH-stretch excitation of the methyl group. Three different vibrational bands are observed, one parallel (k=0<--0) and two perpendicular (/k/1<--0), which for a nearly decoupled methyl rotor framework would correspond to symmetric and (nearly degenerate) asymmetric CH-stretch excitations. However, the splitting between the two asymmetric CH stretch excitations is anomalously large (approximately 125 cm(-1)), signaling the presence of interactions between the CH2 radical moiety and the opposing CH bond on the methyl group. This suggests an improved zeroth-order vibrational description as an isolated CH stretch, strongly redshifted by hyperconjugation, with localized vibrations in the remaining CH bonds split into symmetric and asymmetric stretches. Such a dynamical picture highlights a remarkably strong coupling between methyl CH-stretch vibrations and C-C torsional geometry and begins to elucidate discrepancies with previous matrix observations. PMID- 16468879 TI - Permanent electric dipole moments of four tryptamine conformers in the gas phase: a new diagnostic of structure and dynamics. AB - Rotationally resolved electronic spectroscopy in the gas phase, in the absence and presence of an applied electric field, has been used to determine the charge distribution of a cross section of the energy landscape of tryptamine (TRA). We report the magnitude and direction of the permanent electric dipole moments of the four TRA conformers GPyout, GPyup, GPhup, and Antiup in their S0 and S1 electronic states. Each dipole moment is unique, providing a powerful new tool for the conformational analysis of biomolecules in the gas phase. A comparison of the results for the different conformers of TRA reveals that the position and orientation of the ethylamine side chain play a major role in determining both the permanent and induced electric dipole moments of the different species in both electronic states. PMID- 16468880 TI - Photofragment coincidence imaging of small I-(H2O)n clusters excited to the charge-transfer-to-solvent state. AB - The photodissociation dynamics of small I-(H2O)n(n=2-5) clusters excited to their charge-transfer-to-solvent (CTTS) states have been studied using photofragment coincidence imaging. Upon excitation to the CTTS state, two photodissociation channels were observed. The major channel (approximately 90%) is a two-body process forming neutral I+(H2O)n photofragments, and the minor channel is a three body process forming I+(H2O)n-1+H2O fragments. Both processes display translational energy [P(ET)] distributions peaking at ET=0 with little available energy partitioned into translation. Clusters excited to the detachment continuum rather than to the CTTS state display the same two channels with similar P(ET) distributions. The observation of similar P(ET) distributions from the two sets of experiments suggests that in the CTTS experiments, I atom loss occurs after autodetachment of the excited [I(H2O)n-]* cluster or, less probably, that the presence of the excess electron has little effect on the departing I atom. PMID- 16468881 TI - Rotational averaging and optimization of laser-induced population transfer in molecules. AB - The dynamics of a molecule subject to a short laser pulse is investigated, with focus on the averaging over initial rotational states and on the optimization of laser parameters for the efficient population transfer between vibrational and electronic states. A relation is established between final-state populations obtained with a fixed orientation and those based on a full treatment of the rotational degrees of freedom. In the short-pulse approximation, rotational averaging amounts to integrating the fixed molecule results over all orientations. The theory is applied to a variety of model systems and verified with numerical calculations using Gaussian pulses. We calculate target state populations with three procedures, optimizing the laser pulse for a fixed orientation without orientational averaging, averaging without changing the laser parameters, and reoptimizing the parameters after averaging. The analysis of the two-level system provides a reference for the order of magnitude of the effects of averaging. The three-level system brings out the relevant role of the geometry of polarization vectors and transition dipoles. The multiphoton excitation of a Morse oscillator shows the importance of taking into account the dependence of resonance frequencies on the laser intensity. Within a proton transfer model we discuss the results obtained with and without chirping and we show that "optimizing after averaging" can be as effective as choosing a more refined pulse shape. PMID- 16468882 TI - Electron-impact ionization of helium clusters close to the threshold: appearance energies. AB - We have investigated the ionization threshold behavior of small helium cluster ions (cluster size n=2-10) formed via electron-impact ionization of neutral helium droplets and derive appearance energies for mass-selected cluster ions using a nonlinear least-square-fitting procedure. Moreover, we report magic numbers in the mass spectrum observed at the electron energy of 70 eV. The apparatus used for the present measurements is a hemispherical electron monochromator combined with a quadrupole mass spectrometer. Our experiment demonstrates that helium clusters are not only exclusively formed via direct ionization above the atomic ionization potential but also indirectly via autoionizing Rydberg states. The present results are compared with previous electron-impact and photoionization results. PMID- 16468883 TI - Theoretical investigation of the dissociation dynamics of vibrationally excited vinyl bromide on an ab initio potential-energy surface obtained using modified novelty sampling and feed-forward neural networks. AB - The reaction dynamics of vibrationally excited vinyl bromide have been investigated using classical trajectory methods on a neural network potential surface that is fitted to an ab initio database of 12 122 configuration energies obtained from electronic structure calculations conducted at the MP4(SDQ) level of theory using a 6-31G(d,p) basis set for the carbon and hydrogen atoms and Huzinaga's (43334334) basis set augmented with split outer s and p orbitals (4332143214) and a polarization f orbital with an exponent of 0.5 for the bromine atom. The sampling of the 12-dimensional configuration hyperspace of vinyl bromide prior to execution of the electronic structure calculations is accomplished by combining novelty-sampling methods, chemical intuition, and trajectory sampling on empirical and neural network surfaces. The final potential is obtained using a two-layer feed-forward neural network comprising 38 and 1 neurons, respectively, with hyperbolic tangent sigmoid and linear transfer functions in the hidden and output layers, respectively. The fitting is accomplished using the Levenberg-Marquardt algorithm with early stopping and Bayesian regularization methods to avoid overfitting. The interpolated potentials have a standard deviation from the ab initio results of 0.0578 eV, which is within the range generally regarded as "chemical accuracy" for the purposes of electronic structure calculations. It is shown that the potential surface may be easily and conveniently transferred from one research group to another. The files required for transfer of the vinyl bromide surface can be obtained from the Electronic Physics Auxiliary Publication Service. Total dissociation rate coefficients for vinyl bromide are obtained at five different excitation energies between 4.50 and 6.44 eV. Branching ratios into each of the six open reaction channels are computed at 24 vibrational energies in the range between 4.00 and 6.44 eV. The distribution of vibrational energies in HBr formed via three-center dissociation from vinyl bromide is determined and compared with previous theoretical and experimental results. It is concluded that the combination of ab initio electronic structure calculations, novelty sampling with chemical intuition and trajectories on empirical analytic surfaces, and feed-forward neural networks provides a viable framework in which to execute purely ab initio molecular-dynamics studies on complex systems with multiple open reaction channels. PMID- 16468884 TI - Benchmarking two-photon absorption with CC3 quadratic response theory, and comparison with density-functional response theory. AB - We present a detailed study of the effects of electron correlation on two-photon absorption calculated by coupled cluster quadratic response theory. The hierarchy of coupled cluster models CCS, CC2, CCSD, and CC3 has been used to investigate the effects of electron correlation on the two-photon absorption cross sections of formaldehyde (CH2O), diacetylene (C4H2), and water (H2O). In particular, the effects of triple excitations on two-photon transition cross sections are determined for the first time. In addition, we present a detailed comparison of the coupled cluster results with those obtained from Hartree-Fock and density functional response theories. We have investigated the local-density approximation, the pure Becke-Lee-Yang-Parr (BLYP) functional, the hybrid Becke-3 parameter-Lee-Yang-Parr (B3LYP), and the Coulomb-attenuated B3LYP (CAM-B3LYP) functionals. Our results show that the CAM-B3LYP functional, when used in conjuction with a one-particle basis-set containing diffuse functions, has much promise; however, care must still be exercised for diffuse Rydberg-type states. PMID- 16468885 TI - The application of a vacuum-ultraviolet Fourier transform spectrometer and synchrotron-radiation source to measurements of bands of NO. VII. The final report. AB - Photoabsorption measurements of NO bands have been made by vacuum-ultraviolet Fourier transform spectrometry with a resolution of 0.12 cm(-1) in the wavelength region of 166.2-196.2 nm. Accurate line positions are obtained for the delta(upsilon,0) bands with upsilon=2, 3, the epsilon(upsilon,0) bands with upsilon=2, 3, and the beta(upsilon,0) bands with upsilon=10,12,14. Absolute term values are found for the corresponding upper levels C(2,3), D(2,3), and B(10,12,14). Accurate rotational line integrated cross sections have also been obtained for the lines in these bands. Integrated cross sections reported in our earlier papers [J. Chem. Phys. 109, 1751 (1998); 112, 2251 (2000); 115, 3719 (2001); 116, 155 (2002); 117, 10621 (2002); 119, 8373 (2003)] have been revised, and the results reported here comprise the delta(upsilon,0) bands with upsilon=0 3, the epsilon(upsilon,0) bands with upsilon=0-3, the beta(upsilon,0) bands with upsilon=6,7,9-12,14, and the gamma(3,0) band. For each band, the band oscillator strength is obtained from the sum of the line strengths of all rotational lines, and these are compared with other published values. PMID- 16468886 TI - Probing mechanistic photochemistry of glyoxal in the gas phase by ab initio calculations of potential-energy surfaces and adiabatic and nonadiabatic rates. AB - In the present work, the wavelength-dependent mechanistic photochemistry of glyoxal in the gas phase has been explored by ab initio calculations of potential energy surfaces, surface crossing points, and adiabatic and nonadiabatic rates. The CHOCHO molecules in S1 by photoexcitation at 393-440 nm mainly decay to the ground state via internal conversion, which is followed by molecular eliminations to form CO, H2CO,H2, and HCOH. Upon photodissociation of CHOCHO at 350-390 nm, intersystem crossing to T1 followed by the C-C bond cleavage is the dominant process in this wavelength range, which is responsible for the formation of the CHO radicals. The C-C and C-H bond cleavages along the S1 pathway are energetically accessible upon photodissociation of CHOCHO at 290-310 nm, which can compete with the S1-->T1 intersystem crossing process. The present study predicts that the C-H bond cleavage on the S1 surface is probably a new photolysis pathway at high excitation energy, which has not been observed experimentally. In addition, the trans-cis isomerization is predicted to occur more easily in the ground state than in the excited states. PMID- 16468887 TI - Local control of the quantum dynamics in multiple potential wells. AB - The driven wave-packet dynamics in potentials exhibiting several potential wells is investigated. Therefore, local-control strategies are employed where the control field is constructed from the system's dynamics at any instant of time. It is shown that particles can be moved successively between various potential minima. Furthermore, results presented indicate that the intuitive local-control scheme allows for the initiation of a clockwise or counterclockwise rotational motion of a model molecular motor. PMID- 16468888 TI - Nanosecond rapid freezing of liquid benzene under shock compression studied by time-resolved coherent anti-Stokes Raman spectroscopy. AB - Nanosecond time-resolved coherent anti-Stokes Raman spectroscopy is used to investigate the shock-induced liquid-solid phase transition and crystallization of liquid benzene. Temporal evolution of the Raman shift of the ring-breathing and C-H stretching modes is investigated. A metastable supercompressed state and a liquid-solid phase transition are observed under shock compression. Time resolved Raman spectra reveal that the liquid state is initially a metastable state and rapidly transforms to the solid state within 25 ns under shock compression at 4.2 GPa. PMID- 16468889 TI - On the unsteady-state species separation of a binary liquid mixture in a rectangular thermogravitational column. AB - This paper investigates the unsteady-state species segregation of binary liquid mixtures in rectangular thermogravitational columns. The analysis leads to a procedure to obtain both molecular and thermal diffusion coefficients from transient separation measurements. Two models are presented: first, an ideal model where buoyancy only depends on temperature and second, a general model where buoyancy also varies with composition. Steady-state measurements are not required regardless of which model is chosen. As a result, the new procedure is faster than steady-state procedures. When either the molecular or thermal diffusion coefficient is known a priori, the other can be obtained without knowledge of fluid properties such as density, viscosity, thermal expansion, and compositional coefficients. PMID- 16468890 TI - Pair-correlation functions and phase separation in a two-component point Yukawa fluid. AB - We investigate the structure of a binary mixture of particles interacting via purely repulsive point Yukawa pair potentials with a common inverse screening length lambda. Using the hypernetted chain closure to the Ornstein-Zernike equations, we find that for a system with "ideal" (Berthelot mixing rule) pair potential parameters for the interaction between unlike species, the asymptotic decay of the total correlation functions crosses over from monotonic to damped oscillatory on increasing the fluid total density at fixed composition. This gives rise to a Kirkwood line in the phase diagram. We also consider a "nonideal" system, in which the Berthelot mixing rule is multiplied by a factor (1+delta). For any delta>0 the system exhibits fluid-fluid phase separation and remarkably the ultimate decay of the correlation functions is now monotonic for all (mixture) state points. Only in the limit of vanishing concentration of either species does one find oscillatory decay extending to r=infinity. In the nonideal case the simple random-phase approximation provides a good description of the phase separation and the accompanying Lifshitz line. PMID- 16468891 TI - An effective pair potential for thermodynamics and structural properties of liquid mercury. AB - The properties of liquid mercury are investigated by using an empirical effective pair potential. Its parameters are determined with the aid of Monte Carlo simulation along the liquid branch of the liquid-vapor coexistence curve. The complexity of the electronic structure of dense metal mercury supposes a state dependence of the interatomic interactions, while no more state dependence is found in the metal-nonmetal transition region. It is shown that the use of this effective potential leads to an accurate description of the structural and thermodynamic properties of the expanded liquid mercury. Then, the melting and freezing phenomena are investigated with that potential. Sharp melting and freezing temperatures are observed at 234 and 169 K, respectively. This large hysteresis loop between freezing and melting is consistent with the experiments for the bulk mercury. PMID- 16468892 TI - Critical behavior of the aqueous electrolytic system 3-methylpyridine+D2O+NaBr. AB - The system 3-methylpyridine(3MP) + water(H2O)+NaBr has been the subject of an intense scientific debate since the work of Jacob et al. [Phys. Rev. E. 58, 2188 (1988)] and Anisimov et al. [Phys. Rev. Lett. 85, 2336 (2000)]. The crossover critical behavior of this system seemed to show remarkable sensitivity to the weight fraction (X) of the ionic impurity NaBr. In the range X< or =0.10 the system displayed Ising behavior and a pronounced crossover to mean-field behavior in the range 0.10< or =X< or =0.16. A complete mean-field behavior was observed at X=0.17, a result that was later attributed to the existence of long-living nonequilibrium states in this system [Kostko et al., Phys. Rev. E. 70, 026118 (2004)]. In this paper, we report the near-critical behavior of osmotic susceptibility in the isotopically related ternary system, 3MP+heavy water(D2O)+NaBr. Detailed light-scattering experiments performed at exactly the same NaBr concentrations as investigated by Jacob et al. reveal that the system 3MP + D2O + NaBr shows a simple Ising-type critical behavior with gamma approximately 1.24 and nu approximately 0.63 over the entire NaBr concentration range 0< or =X< or =0.1900. The crossover behavior is predominantly nonmonotonic and is completed well outside the critical domain. An analysis in terms of the effective susceptibility exponent (gammaeff) reveals that the crossover behavior is nonmonotonic for 0< or =X< or =0.1793 and tends to become monotonic for X>0.1793. The correlation length amplitude xio, has a value of approximately 2 A for 0.0250< or =X< or =0.1900, whereas for X=0, xio approximately 3.179 A. Since isotopic H-->D substitution is not expected to change the critical behavior of the system, our results support the recent results obtained by Kostko et al. [Phys. Rev. E. 70, 026118 (2004)] that 3MP + H2O + NaBr exhibits universal Ising type critical behavior typical for other aqueous solutions. PMID- 16468893 TI - Theory of electron solvation in polar liquids: a continuum model. AB - The solvation of electrons in polar liquids is analyzed on the basis of an extended continuum model. In addition to the long-range electron-dipole interaction two short-range interactions are introduced. Among others one accounts for interactions with groups capable of forming hydrogen bonds and the second for quadrupolar characteristics of the liquid molecules. Both are induced by the orientation of the molecular dipole. Applying the scaling method a proper reaction coordinate is introduced and the solvation dynamics are discussed for the electron in the electronic ground state and after excitation to the p-type excited state. The observed spectral evolution of the transient absorption spectra, after two photon excitations for electrons in water and in methanol, is well described by this theory. An analytic estimate for the nonradiative deactivation from the electronically excited solvated electron is found to be consistent with an observed lifetime of 50 fs for the electron in water. The theory predicts an about three times slower internal conversion in methanol as solvent in comparison with water. PMID- 16468894 TI - Molecular-dynamics simulation of argon nucleation from supersaturated vapor in the NVE ensemble. AB - The possibility to conduct simulations of homogeneous nucleation of argon from a supersaturated vapor phase using a microcanonical or NVE ensemble is evaluated (NVE: number of particles N, volume V, and energy E are constant). In order to initiate a phase separation kinetic energy is removed from the system in one step which transfers the system into a supersaturated state. After this temperature jump the simulation is continued in a NVE ensemble. The simulations are performed for different initial-state points and different temperature jumps. The cluster formation and growth over the course of the adiabatic simulations are analyzed. The progression of the temperature being related to the cluster size in NVE systems is traced. Also the influence of the size of the simulation system is investigated. For a certain range of low supersaturation a dynamic coexistence between two states has been found. Furthermore, the obtained nucleation rates are correlated with two simple functions. By applying the nucleation theorems to these functions the size and excess energy of the critical cluster are estimated. The results are consistent with other theoretical data and experimental data available in the literature. PMID- 16468895 TI - The chemisorption of coronene on Si(001)-2 x 1. AB - Coronene (C24H12) adsorption on the clean Si(001)-2 x 1 surface was investigated by scanning tunneling microscopy and by density-functional calculations. The coronene adsorbed randomly at 25 degrees C on the surface and did not form two dimensional islands. The scanning tunneling microscopy measurements revealed three adsorption sites for the coronene molecule on the Si(001) surface at low coverage. The major adsorption configuration involves coronene bonding to four underlying Si atoms spaced two lattice spacings apart in a dimer row. The two minor adsorption configurations involve asymmetrical bonding of a coronene molecule between Si dimer rows and form surface species with a mirror plane symmetry to their chiral neighbor species. The two minor bonding arrangements are stabilized by a type-C defect on the Si(001) surface. PMID- 16468896 TI - Dynamic contact angle in rim instability of dewetting holes. AB - The effects of dynamic contact angle (thetad), between a substrate and the melt of a dewetting polymer thin film, on the evolution of rim instabilities of dewetting holes were reported. Various thetad's were achieved by covering SiOx surfaces with different coverage of octadecyltrichlorosilane. On each surface, the morphology of the dewetting holes was examined in detail as the hole grew to a certain size. Rim instabilities, in terms of undulations in both r and z directions, became more pronounced as thetad increased, under which condition, narrower and higher rims were also observed. Experimentally, atomic force microscopic scans of the rim were used to obtain the rim profile, which was predicted using thetad. The predicted rim profile was used, in combination with the analysis of Rayleigh instability of a cylindrical fluid, to interpret the rim instability. The model captures the basic trend of the rim instability dependency on thetad. The study demonstrates the importance of the substrate properties on the rim instability and the destabilization of polymer thin films during hole growth. PMID- 16468897 TI - Rheo-optical studies of carbon nanotube suspensions. AB - We use a polarization-modulation technique to investigate the optical anisotropy of multi- and single-wall carbon nanotubes suspended in a variety of solvents under simple shear flow. Measurements of birefringence and dichroism are performed as a function of shear rate, tube concentration, and solvent viscosity. At fixed volume fraction, the anisotropy increases with increasing shear stress due to enhanced flow alignment. At fixed shear stress, the anisotropy increases with volume fraction due to rotational excluded-volume interactions. By considering the rotational diffusivity as a function of nanotube length, diameter, concentration, and solvent viscosity, we demonstrate a leading-order scaling relation for the optical anisotropy in terms of rotary Peclet number Pe. At low Pe, our results are in qualitative agreement with the theoretical predictions of Doi and Edwards. At high Pe, our data suggest that the degree of nanotube alignment scales as Pe16. PMID- 16468898 TI - Structural characterizations and electronic properties of Ti-doped SnO2(110) surface: a first-principles study. AB - The Ti-doped SnO2(110) surface has been investigated by using first-principles method with a slab model. The geometrical optimizations and band-structure calculations have been performed for four possible doping models. Our results indicate that the substitution of Ti for sixfold-coordinated Sn atom at the top layer is most energetically favorable. Compared to the undoped surface, those Sn and O atoms located above Ti atom tend to move toward the bulk side. Besides the surface relaxations, the doping of Ti has significant influences on the electronic structures of SnO2(110) surface, including the value and position of minimum band gap, the components of valence and conduction bands, the distributions of the charge densities, and the work function of the surface. Furthermore, the effects introduced by the substitution of Ti atom observed in the experiments can be well explained when the sixfold-coordinated Sn atom at the first layer is replaced by Ti atom. PMID- 16468899 TI - Cn films (n=50, 52, 54, 56, and 58) on graphite: cage size dependent electronic properties. AB - Novel semiconducting materials have been prepared under ultrahigh-vacuum conditions by soft-landing mass-selected Cn+ (50< or =n<60; even n) on highly oriented pyrolytic graphite surfaces at mean kinetic energies of 6 eV. In all cases, Cn films grow according to the Volmer-Weber mechanism: the surface is initially decorated by two-dimensional fractal islands, which in later deposition stages become three-dimensional dendritic mounds. We infer that Cn aggregation is governed by reactive sites comprising adjacent pentagons (or heptagons) on individual cages. The resulting covalent cage-cage bonds are responsible for the unusually high thermal stability of the films compared to solid C60. The apparent activation energies for intact Cn sublimation range from 2.2 eV for C58 to 2.6 eV for C50 as derived from thermal desorption spectra. All Cn films exhibit a common valence-band ultraviolet photoelectron spectroscopy spectral feature located around the center of a broad highest occupied molecular-orbital (HOMO)-derived band (EB approximately 2.5 eV). This feature has been assigned to Cn units covalently linked to each other in polymeric structures. To within experimental accuracy, the same work function (4.8 eV) was determined for thick films of all Cn studied. In contrast, "HOMO" ionization potentials were cage size dependent and significantly lower than that obtained for C60. C58 exhibited the lowest HOMO (6.5 eV). Band gaps of Cn films have been determined by depositing small amounts of Cs atoms onto the topmost film layer. HOMO-lowest unoccupied molecular-orbital derived band gaps between 0.8 eV (C52) and 1.8 eV (C50) were observed, compared to 1.5 eV for solid C60. PMID- 16468900 TI - Defects-enhanced dissociation of H2 on boron nitride nanotubes. AB - With the density-functional theory and nudged elastic band method, the adsorption and dissociation of the hydrogen molecule on the boron nitride (BN) nanotubes with and without defects are studied theoretically. Hydrogen molecule physically adsorbs on the surface of the BN layer and nanotubes. The dissociation of the hydrogen molecule on the surface of the perfect BN layer and nanotubes is endothermic, and the energy barrier reduces with the decrease of the diameter of the tubes, while it is still larger than 2.0 eV for the (7,0) BN nanotube. Antisite, carbon substitutional, vacancy, and Stone-Wales 5775 defects on the wall of the tube are considered. With the presence of the defects, the dissociation of the hydrogen molecule becomes exothermic and the dissociation barrier can be reduced to about 0.67 eV. PMID- 16468901 TI - Finite-element implementation for electron transport in nanostructures. AB - We have modeled transport properties of nanostructures using Green's-function method within the framework of the density-functional theory. The scheme is computationally demanding, so numerical methods have to be chosen carefully. A typical solution to the numerical burden is to use a special basis-function set, which is tailored to the problem in question, for example, the atomic-orbital basis. In this paper we present our solution to the problem. We have used the finite-element method with a hierarchical high-order polynomial basis, the so called p elements. This method allows the discretation error to be controlled in a systematic way. The p elements work so efficiently that they can be used to solve interesting nanosystems described by nonlocal pseudopotentials. We demonstrate the potential of the implementation with two different systems. As a test system a simple Na-atom chain between two leads is modeled and the results are compared with several previous calculations. Secondly, we consider a thin hafnium dioxide (HfO2) layer on a silicon surface as a model for a gate structure of the next generation of microelectronics. PMID- 16468902 TI - Adsorption and diffusion of carbon dioxide and nitrogen through single-walled carbon nanotube membranes. AB - We have used atomically detailed simulations to examine the adsorption and transport diffusion of CO2 and N2 in single-walled carbon nanotubes at room temperature as a function of nanotube diameter. Linear and spherical models for CO2 are compared, showing that representing this species as spherical has only a slight impact in the computed diffusion coefficients. Our results support previous predictions that transport diffusivities of molecules inside carbon nanotubes are extremely rapid when compared with other porous materials. By examining carbon nanotubes as large as the (40,40) nanotube, we are able to compare the transport rates predicted by our calculations with recent experimental measurements. The predicted transport rates are in reasonable agreement with experimental observations. PMID- 16468903 TI - Quantum chemical modeling of electrochromism of tungsten oxide films. AB - A cluster model is proposed to describe the excitations in solid tungsten oxide. The density-functional theory approach is used to calculate the ground-state electronic structure of the model cluster and its optimum geometry; subsequently, time-dependent density-functional theory calculations are performed to obtain the oscillator strengths and energies of the excited states. The results are reported both for the electrically neutral cluster and for the cluster with an extra electron (mimicking the effect of electron injection from the cathode). They correctly locate the electrochemically active transition. The corresponding wave functions are delocalized, suggesting that electron localization at one tungsten center is rather unlikely, thereby shedding doubt as to the validity of the polaron model. Local lattice distortions presumably created at the stage of sample preparation are found to affect the excitation energies to a considerable extent, which explains the experimentally observable large width of optical absorption responsible for electrochromism. PMID- 16468904 TI - Isothermal desorption measurements of self-diffusion in supercooled o-terphenyl. AB - Isothermal desorption of o-terphenyl thin-film bilayers was used to measure self diffusion coefficients of supercooled o-terphenyl near the glass transition temperature (Tg=243 K). Diffusion coefficients from 10(-15.5) to 10(-12) cm2 s( 1) were obtained between 246 and 265 K. Protio and deuterio o-terphenyl were sequentially vapor deposited, then annealed to simultaneously diffuse and desorb the sample in a vacuum chamber. During the desorption of the bilayer, the concentration of each isotope was detected by a mass spectrometer, which revealed the extent of interfacial broadening. In these experiments, isotopic interdiffusion is indistinguishable from self-diffusion and the measured interfacial broadening is consistent with Fickian diffusion. The samples prepared under several different deposition conditions yielded the same self-diffusion coefficients, indicating that the experiments were conducted in the equilibrium supercooled liquid state. PMID- 16468905 TI - Crystallographic and morphological characterization of thin pentacene films on polycrystalline copper surfaces. AB - The degree of crystallinity, the structure and orientation of crystallites, and the morphology of thin pentacene films grown by vapor deposition in an ultrahigh vacuum environment on polycrystalline copper substrates have been investigated by x-ray diffraction and tapping-mode scanning force microscopy (TM-SFM). Depending on the substrate temperature during deposition, very different results are obtained: While at 77 K a long-range order is missing, the films become crystalline at elevated temperatures. From a high-resolution x-ray-diffraction profile analysis, the volume-weighted size of the crystallites perpendicular to the film surface could be determined. This size of the crystallites increases strongly upon changing temperature between room temperature and 333 K, at which point the size of individual crystallites typically exceeds 100 nm. In this temperature region, three different polymorphs are identified. The vast majority of crystallites have a fiber texture with the (001) net planes parallel to the substrate. In this geometry, the molecules are oriented standing up on the substrate (end-on arrangement). This alignment is remarkably different from that on single-crystalline metal surfaces, indicating that the growth is not epitaxial. Additionally, TM-SFM images show needlelike structures which suggest the presence of at least one additional orientation of crystallites (flat-on or edge-on). These results indicate that properties of thin crystalline pentacene films prepared on technologically relevant polycrystalline metal substrates for fast electronic applications may be compromised by the simultaneous presence of different local molecular aggregation states at all temperatures. PMID- 16468906 TI - Anomalous potentials from inverse analyses of interfacial polydisperse attractive colloidal fluids. AB - This paper investigates effects of using monodisperse inverse analyses to extract particle-particle and particle-surface potentials from simulated interfacial colloidal fluids of polydisperse attractive particles. Effects of polydispersity are investigated as functions of particle concentration and attractive well depth and range for van der Waals and depletion potentials. Forward Monte Carlo simulations are used to generate particle distribution functions for polydisperse interfacial colloidal fluids from which inverted potentials are obtained using an inverse Ornstein-Zernike analysis and an inverse Monte Carlo simulation method. Attractive potentials are successfully recovered for monodisperse colloidal fluids, but polydispersity that is unaccounted for in inverse analyses produces (1) apparent softening of strong forces, (2) anomalous repulsive and attractive interactions, and (3) aphysical particle overlaps. This investigation provides insights into the role of polydispersity in altering the equilibrium structure and corresponding inverted potentials of attractive colloidal fluids near surfaces. These findings should assist the design and interpretation of optical microscopy experiments involving interfacial colloidal fluids similar to the simulated experiments reported here. PMID- 16468907 TI - The effect of impurities on jamming in random sequential adsorption of elongated objects. AB - We consider the jamming aspect of random sequential adsorption of extended particles onto two-dimensional lattice by computer Monte Carlo simulations. The initial presence of impurities on the substrate disturbs this phenomenon significantly and we study here how the size and density of impurity particles affect the resulting jamming threshold. We present the formula for jamming threshold as a closed function of all important parameters (the size of primary particles, the size of impurity particles, and the final density of impurities). The fractal dimension of the space free of impurities is also discussed. PMID- 16468908 TI - Towards understanding ice nucleation by long chain alcohols. AB - We demonstrate that infrared spectra of water covered by a film of heptadecanol show a continuous spectral shift, from a band characteristic of liquid water to one characteristic of ice, as the temperature is ramped from -10 to -17 degrees C. Experiments with pure water and water covered by films of long chain alkanes showed no such spectral shift. Analysis of the CH2 stretching features in the alcohols' absorbance bands reveals simultaneous structural changes within the alcohol film. We hypothesize that the spectral shift in the water band is due to an increasing fraction of water molecules participating in icelike clusters and that these clusters are stabilized by the influence of the flexible alcohol film. PMID- 16468909 TI - Angular distributions of H-induced HD and D2 desorptions from the Si(100) surfaces. AB - We measured angular distributions of HD and D2 molecules desorbed via the reactions H+DSi(100)-->HD [abstraction (ABS)] and H+DSi(100)-->D2 [adsorption induced-desorption (AID)], respectively. It was found that the angular distribution of HD molecules desorbed along ABS is broader than that of D2 molecules desorbed along AID, i.e., the former could be fit with cos(2.0+/-0.2) theta, while the latter with cos(5.0+/-0.5) theta. This difference of the angular distributions between the two reaction paths suggests that their dynamic mechanisms are different. The observed cos2 theta distribution for the ABS reaction was reproduced by the classical trajectory calculations over the London Eyring-Polanyi-Sato potential-energy surfaces. The simulation suggests that the HD desorption along the ABS path takes place along the direction of Si-D bonds, but the apparent angular distribution is comprised of multiple components reflecting the different orientations of D-occupied Si dimers in the (2 x 1) and (1 x 2) double domain structures. PMID- 16468910 TI - The ordered thin-film growth of organic semiconductor on Ag(110). AB - Growth of ordered perylene thin films on the Ag(110) surface has been investigated with scanning tunneling microscope. By saturating the surface with a monolayer of perylene molecules, two kinds of ordered structures are simultaneously formed with flat-lying perylene molecules on the Ag(110) surface, in which one is commensurate relative to the Ag substrate with a periodicity of while the other is commensurate with a periodicity of (-2724). There is one molecule within the former unit cell with a surface coverage of 0.1 molecule per Ag atom, while there are two molecules within the latter unit cell, which gives a slightly lower surface coverage of 0.091 molecule per Ag atom. Ab initio calculations have been carried out to identify the adsorption geometry and bonding sites. PMID- 16468911 TI - Simultaneous in situ monitoring of surface and gas species and surface properties by modulation excitation polarization-modulation infrared reflection-absorption spectroscopy: CO oxidation over Pt film. AB - A method for in situ monitoring of surface and gas species utilizing separately the difference and sum reflectivity of two polarizations, normal and parallel to the surface, measured by polarization-modulation infrared reflection-absorption spectroscopy is presented. Surface and gas-phase spectra were separately but simultaneously obtained from the reflectivities. The technique is combined with modulation excitation spectroscopy to further enhance the sensitivity, and a small-volume cell was designed for this purpose. CO oxidation over a 40 nm Pt film on aluminum was investigated under moderate pressure (atmospheric pressure, 5% CO, and 5%-40% O2) at 373-433 K. The surface species involved in the oxidation process and the gas-phase species, both reactant (CO) and product (CO2), could be simultaneously monitored and analyzed quantitatively. In addition, the reflectivity change of the sample during the reaction was assigned to a near surface bulk property change, that is, surface reconstruction to the oxide phase. Under an O2-rich atmosphere, two reactive phases, denoted as low- and high activity phases, were identified. A large amount of atop CO was observed during the low-activity phase, while the adsorbed CO completely disappeared during the high-activity phase. The presence of an infrared-inactive CO2 precursor formed by the reaction between surface oxide and gaseous CO during the high-activity phase was inferred. The desorption of the CO2 precursor is facilitated under a CO-rich atmosphere, most likely, by surface reconstruction to metallic Pt and a competitive adsorption of CO on the surface. PMID- 16468912 TI - Molecular dynamics in n-alkanes: premelting phenomena and rotator phases. AB - Molecular dynamics simulations of the n-alkanes C18H38, C19H40, and C20H42 are reported for temperatures just below the melting point. Besides thermodynamic and average structural data for the ordered phase, we discuss the molecular motions initiating the rotator phases observed in spontaneous phase transitions in isothermal, isostress simulations. The RI phase of C19H40 is initiated by particular cork-screw-like jumps combining a quarter turn about the long molecular axis and a half-chain-period translation along the axis. This motion occurs between the minimum-energy conformation of the ordered crystal and a secondary minimum. Transient analogs of the RI and RII phases of the odd alkanes are found on melting C18H38 and C20H42. Collective motions within lamellae of molecules are prominent in the dynamics. PMID- 16468913 TI - Shear-induced parallel-to-perpendicular orientation transition in the amphiphilic lamellar phase: a nonequilibrium molecular-dynamics simulation study. AB - The present work is devoted to a study of the shear-induced parallel-to perpendicular orientation transition in the lamellar system by the large-scale nonequilibrium molecular-dynamics (NEMD) simulation. An effective generic model A2B2 tetramer for amphiphilies is used. The NEMD simulation produces unambiguous evidence that undulation instability along the vorticity direction sets in well above a critical shear rate and grows in magnitude as the shear rate is further increased. At a certain high shear rate, the coherent undulation instability grows so large that defects are nucleated and the global lamellar monodomain breaks into several aligned lamellar domains. Subsequently layers in these domains rotate into the perpendicular orientation with the rotation of chains towards the y direction, merge into a global perpendicular-aligned lamellar monodomain, and organize into a perfect well-aligned perpendicular lamellar phase by the migration and annihilation of edge dislocations and disclinations. The macroscopic observable viscosity as a function of time or shear rate is correlated with the structural response such as the mesoscopic domain morphology and the microscopic chain conformation. The onset of undulation instability concurs with the start-up of shear-thinning behavior. During the orientation transformation at the high shear rate, the complex time-dependent thixotropic behavior is observed. The smaller viscosity in the perpendicular lamellar phase gives an energetic reason for the shear-induced orientation transition. PMID- 16468914 TI - Control of structure and photophysical properties by protonation and subsequent intramolecular hydrogen bonding. AB - Protonation and subsequent intramolecular hydrogen bonding as methods to control chain structure and tune luminescence in heteroatomic conjugated polymers were reported experimentally [A. P. Monkman et al., J. Am. Chem. Soc. 124, 6049 (2002)]. In this paper, the structure and photophysical properties of the model teraryl compound of phenylene-pyridylene copolymer before and after protonation are theoretically studied with quantum chemistry methods. From the optimized ground states, intramolecular hydrogen bonding to the adjacent oxygen atom in the alkoxy substituent planarizes the backbone of the molecules, and the optimized detailed results of compound 9 before and after protonation, such as the dihedral angles between the central benzene and the two pyridyl rings, the bond lengths, and the bond angles, are consistent with the experimental results. From the results of the calculated excited states, the protonation and subsequent intramolecular hydrogen bonding result in the redshifts of the absorption, the increase of the ionization energy, the increase of the electron affinity, the decrease of the energy difference of the highest occupied molecular orbital and lowest unoccupied molecular orbital, the decrease of the binding gap, and the delocalization of the electron-hole coherence. The photophysical properties of compound 9 before and after protonation are further studied with a three dimensional real-space analysis method of transition and charge difference densities (study transition dipole moment and charge transfer in the absorption and fluorescence processes) and two-dimensional real-space analysis method of transition density matrices (study the electron-hole coherence and the excitation delocalization). The calculated results show theoretically an insight understanding on the influence of the protonation and subsequent intramolecular hydrogen bonding to chain structure and photophysical properties. PMID- 16468915 TI - Polyene photoisomerization rates: are they distinct in aqueous block copolymer micellar solutions and gels? AB - Photoisomerization of 3,3'-diethyloxadicarbocyanine iodide (DODCI) has been investigated in water, 5% and 30% aqueous triblock copolymer, poly(ethylene oxide)20-poly(propylene oxide)70-poly(ethylene oxide)20 (P123) by measuring the fluorescence quantum yields and lifetimes in the temperature range 293-318 K. Reports available in literature indicate that 5% aqueous P123 exists as micellar solution, whereas 30% aqueous P123 forms gel due to micelle-micelle entanglement. This study has been undertaken to find out how the polyene photoisomerization rates are influenced in the sol and gel phases. It has been observed that 60%-70% of DODCI is located in the palisade layer of the micelles in the sol as well as gel phases and the photoisomerization rate of this component is identical in both the phases at a particular temperature. The remainder of the probe is located in the interfacial region and isomerization rates of this fraction are slower by a factor of 1.4-1.1 in the gel phase compared with the micellar solution. The retardation of the isomerization rate in the gel phase has been explained on the basis of enhancement in the friction experienced by the probe due to micelle micelle entanglement at the interface. Compared to the isomerization rates in water, the rates of photoisomerization of DODCI located in the palisade layer, interfacial region of micellar solution, and interfacial region of the micelles in the gel phase are slower by factors of 3.5, 1.5-1.9, and 2, respectively. The outcome of this study validates the point that in organized media photoisomerization rates are sensitive to the localized friction, which is not uniform unlike in a homogeneous solution. PMID- 16468916 TI - Computing the starting state for Gibbs-Duhem integration. AB - Gibbs-Duhem integration implies the numerical integration of a Clapeyron equation. To start the numerical integration, an initial coexistence point and a corresponding initial slope of the Clapeyron equation are needed. In order to apply Gibbs-Duhem integration to all kinds of systems at diverse physical conditions, one has to investigate and assess the available methods that can be used to compute these initial values. This publication focuses on vapor-liquid equilibria in binary mixtures comprising chain molecules. The initial coexistence point is either computed with the NVbeta Gibbs ensemble or with the Npbeta+test molecule method with overlapping distributions, which is introduced in this publication. Although computationally demanding, the Npbeta+test molecule method with overlapping distributions is applicable at conditions where the NVbeta Gibbs ensemble fails. We investigated three methods that can be employed to compute the initial slope of the Clapeyron equation. The Widom method and the overlapping distributions difference method provide correct values for the initial slope. The difference method does only provide the correct answer in special cases. The possibility to judge the reliability of the results makes the overlapping distributions difference method the safest route to the initial slope. Gibbs Duhem integration requires the frequent computation of the slope of the Clapeyron equation. This slope depends on ensemble averages of the composition. A new bias method for efficient sampling of the composition in a semigrand-canonical simulation of chain molecules is presented. This bias method considerably enhances the composition sampling in systems comprising chain molecules of different sizes. PMID- 16468917 TI - An advanced Gibbs-Duhem integration method: theory and applications. AB - The conventional Gibbs-Duhem integration method is very convenient for the prediction of phase equilibria of both pure components and mixtures. However, it turns out to be inefficient. The method requires a number of lengthy simulations to predict the state conditions at which phase coexistence occurs. This number is not known from the outset of the numerical integration process. Furthermore, the molecular configurations generated during the simulations are merely used to predict the coexistence condition and not the liquid- and vapor-phase densities and mole fractions at coexistence. In this publication, an advanced Gibbs-Duhem integration method is presented that overcomes above-mentioned disadvantage and inefficiency. The advanced method is a combination of Gibbs-Duhem integration and multiple-histogram reweighting. Application of multiple-histogram reweighting enables the substitution of the unknown number of simulations by a fixed and predetermined number. The advanced method has a retroactive nature; a current simulation improves the predictions of previously computed coexistence points as well. The advanced Gibbs-Duhem integration method has been applied for the prediction of vapor-liquid equilibria of a number of binary mixtures. The method turned out to be very convenient, much faster than the conventional method, and provided smooth simulation results. As the employed force fields perfectly predict pure-component vapor-liquid equilibria, the binary simulations were very well suitable for testing the performance of different sets of combining rules. Employing Lorentz-Hudson-McCoubrey combining rules for interactions between unlike molecules, as opposed to Lorentz-Berthelot combining rules for all interactions, considerably improved the agreement between experimental and simulated data. PMID- 16468918 TI - Microphase separation in multiblock copolymer melts: Nonconventional morphologies and two-length-scale switching. AB - The phase behavior of AfmN(BN2AN2)B(1-fmN) multiblock copolymer melts is studied within the weak segregation theory. The interplay between ordering on different length scales is shown to cause dramatic changes both in the ordered phase symmetry and periodicity upon small variation of the architectural parameters of the macromolecules. Phase diagrams are presented in the (f,chiN) plane (chi is the Flory-Huggins parameter) for various values of the architecture parameters n and m. Near the critical surface, i.e., for (f-0.5)2<<1, such nonconventional cubic phases as the face-centered cubic (FCC), simple cubic (SC), (double) gyroid, and the so-called BCC(2) (single gyroid) are found to be stable. The lamellar morphology is shown to be replaced by BCC2, FCC, or SC (depending on the structural parameters) as the most stable low-temperature phase. PMID- 16468919 TI - Molecular mass and dynamics of poly(methyl acrylate) in the glass-transition region. AB - The segmental dynamics of bulk poly(methyl acrylate) (PMA) were studied as a function of molecular mass in the glass-transition region using 2H NMR and modulated differential scanning calorimetry (MDSC). Quadrupole-echo 2H NMR spectra were obtained for four samples of methyl-deuterated PMA-d3 with different molecular masses. The resulting spectra were fit using superpositions of simulated spectra generated from the MXQET simulation program, based on a model incorporating nearest-neighbor jumps from positions on the vertices of a truncated icosahedron (soccer-ball shape). The lower molecular-mass samples, influenced by the presence of more chain ends, showed more heterogeneity (broader distribution) and lower glass transitions than the higher molecular-mass samples. The MDSC experiments on both protonated and deuterated samples showed behavior consistent with the NMR results, but temperature shifted due to the different frequency range of the measurements in terms of both the position and breadth of the glass transition as a function of molecular mass. PMID- 16468920 TI - Liquid-gas separation in colloidal electrolytes. AB - The liquid-gas transition of an electroneutral mixture of oppositely charged colloids, studied by Monte Carlo simulations, is found in the low-temperature-low density region. The critical temperature shows a nonmonotonous behavior as a function of the interaction range, kappa(-1), with a maximum at kappasigma approximately 10, implying an island of coexistence in the kappa-rho plane. The system is arranged in such a way that each particle is surrounded by shells of particles with alternating charge. In contrast with the electrolyte primitive model, both neutral and charged clusters are obtained in the vapor phase. PMID- 16468921 TI - Editorial. PMID- 16468922 TI - Biomolecular applications of carbon nanotubes. AB - Carbon nanotubes are a significant addition to the emerging field of nanotube biotechnology. The biocompatibility, high structural integrity, and unique electronic and mechanical properties of carbon nanotubes complement or surpass those of self-assembled lipid nanotubes, peptide nanotubes, and template synthesised nanotubes (metals, polymers, semiconductors, and carbons). Carbon nanotubes are candidates for a range of biomolecular applications that is likely to widen considerably in the future. PMID- 16468923 TI - Implementation of a redox microarray: an experimental model for future nanoscale biomolecular computing using integrated circuits. AB - The possibility of constructing high-density parallel computing architectures using molecular electronics technology is explored. By employing molecular computing devices, new circuitsystem integration could be realised. To clarify the proposed concept, an experimental model of a redox microarray is presented. A first experimental system for a redox microarray consists of a two-dimensional array of platinum microelectrodes to catalyse reversible reactions of redox active molecules. Experimental results of active wave propagation in the redox microarray are presented to demonstrate the potential of molecular computing devices for creating artificially programmable reaction-diffusion dynamics for specific target applications. PMID- 16468924 TI - Computer-controlled piezoactuator for cell manipulation. AB - The development of a computer-controlled piezomanipulation system is presented for biomedical applications involving cells manipulation. An application of the developed system to intracytoplasmic sperm injection (ICSI) is presented. The hardware setup and the control strategies are described in full detail to illustrate the advantages of this approach compared to manual injection methods. The system developed offers the automated and efficient execution of the ICSI process, thus achieving an improved success rate in terms of the end objectives of the process. PMID- 16468925 TI - Theoretical study of stability problems for transport in a deformable microfabricated channel. AB - We investigate the stability problem related to the basic flows induced by peristaltic transport within the bounded deformable walls which are common in micro- andor nanobiotechnological applications. The neutral stability boundary is obtained by solving the relevant Orr-Sommerfeld equation via a verified preconditioned complex-matrix solver. The critical Reynolds number (when the wall is deformable) is 2886.5 which is much less than the conventional rigid-wall case ( approximately 5772, obtained by Orszag based on the spectral method). PMID- 16468926 TI - Planar silicon nitride waveguides for biosensing. AB - The principles of attenuation of the light intensity due to multiple reflections are realised in a planar silicon oxide (SiO(2))silicon nitride (Si(3)N(4)) waveguiding structure for the purpose of developing optical biosensors with improved sensitivity. The analysis of the experimental data shows that the large difference in refractive indices of core and cladding layers gives rise to an increase in sensitivity by a factor of 3 over previously reported structures. Composite polyelectrolyte self-assembled thin films containing cyclo-tetra chromotropylene as an indicator and enzymes glucose oxidase or urease were employed in the superstrate as a sensing membrane. Individual enzyme reactions as well as their inhibition by pesticides were studied by monitoring the intensity of light output from the planar waveguide. The results were compatible with those obtained by conventional ultraviolet-visible absorption spectroscopy. The instrument detection limit for Imidacloprid pesticide was found to be as low as 10 ppb in concentration. PMID- 16468927 TI - Preliminary electrochemical characterisation of cytochrome P4501A2-clozapine interaction. AB - Cytochromes P450 are a large superfamily of heme-thiolate enzymes involved in the metabolism of many different organic substrates such as drugs, fatty acids and toxic compounds. The aim of this work is to analyse the binding between the cytochrome P4501A2, in solution and in gel-matrix, and its substrate (clozapine), utilising voltammetric tests. The interaction measurements were carried out using two different screen printed electrodes (rhodium-graphite and graphite riboflavin), and the results were compared. It was demonstrated that it is possible to realise a biosensor prototype to detect the presence of clozapine indirectly by chronoamperometry. PMID- 16468928 TI - Editorial: AC electrokinetics of molecules and macromolecular assemblies. PMID- 16468929 TI - Influence of scale on electrostatic forces and torques in AC particulate electrokinetics. AB - Dielectrophoretic forces and torques move and manipulate biological cells, typically of the order of 10 mum ( approximately 10(-5) m) in diameter and ordinarily suspended in aqueous liquids, using electrodes with dimensions around 100 mum ( approximately 10(-4) m). The ability to exploit these same electromechanical effects for particles below 1 mum, that is, <10(-6) m, creates opportunities for remote manipulation and handling of subcellular components, biological macromolecules, and DNA. In this paper, Trimmer's bracket notation is adapted for systematic examination of the scaling laws governing electrokinetic behaviour. The purpose is to shed light on how critical performance measures relevant to the laboratory on a chip are affected by reducing particle sizes and electrode dimensions into the nanometre range. The scaling methodology facilitates consideration of the effect of electrode structure and particle size reduction on voltage, electric field, heating, and response time. Particles with induced moments, dipolar and quadrupolar, as well as permanent dipoles are examined. Separate consideration is given to electrical torque and its application in electrorotation and particle alignment. An eventual goal of these scaling studies is to identify the lower limit on the size of particles that can be manipulated effectively using electrokinetic phenomena. PMID- 16468930 TI - Dielectrophoretic manipulation of DNA. AB - The characterisation and spatial manipulation of cells by AC electrokinetic methods such as dielectrophoresis and electrorotation is well established. However, applications to submicroscopical objects like viruses and molecules have been rare. Only recently has the number of such studies risen more quickly due to the availability of suitable electrodes and a growing need for single molecule techniques. Of special interest is the spatial control of single DNA molecules for genetic investigations as well as for the building of well defined structures with nanometre resolution. Here a review is given of dielectrophoretic studies dealing with single and double stranded DNA emphasising single molecule aspects. PMID- 16468931 TI - Dielectrophoretic manipulation of surface-bound DNA. AB - Dielectrophoretic manipulation enables the positioning and orientation of DNA molecules for nanometer-scale applications. However, the dependence of the dielectrophoretic force and torque on the electric field magnitude and frequency has to be well characterised to realise fully the potential of this technique. DNA in solution is attracted to the strongest electric field gradient (i.e. the electrode edge) as a result of the dielectrophoretic force, while the dielectrophoretic torque aligns the DNA with its longest axis parallel to the electric field. In this work, the authors attached -DNA fragments (48 and 25 kilobases) to an array of gold microelectrodes via a terminal thiol bond and characterised the orientation and elongation as a function of electric field magnitude (0.1-0.8 MVm) and frequency (0.08-1.1 MHz). Maximum elongation was observed between 200 and 500 kHz for the attached DNA. Dielectrophoresis is limited by thermal randomisation at electric fields below 0.1 MVm and by electrothermal effects above 0.7 MVm. The authors conclude that dielectrophoresis can be used to manipulate surface-immobilised DNA reproducibly. PMID- 16468932 TI - Dielectrophoresis of microbioparticles in water with planar and capillary quadrupole electrodes. AB - Dielectrophoresis of single microbioparticles was measured in a planar quadrupole microelectrode (50 mum or 65 mum in working area radius) with a microscope. Carbon and polystyrene microparticles, yeast cells and DNA molecules (about 40 kbp) were adopted as a sample. Their dielectrophoretic mobilities were analysed quantitatively with their intrinsic and surface conductivity, their permittivities and their sizes as well as the conductivity and permittivity of aqueous media. Using the dielectrophoretic mobilities obtained with the planar quadrupole microelectrode, some instances of the separation performance between the microparticles were demonstrated with a fabricated capillary quadrupole microelectrode (82.5 mum in bore radius) under the field flow fractionation regime. PMID- 16468933 TI - Separation of latex spheres using dielectrophoresis and fluid flow. AB - The authors present a method for separation of two latex spheres populations using dielectrophoresis (DEP) and the fluid drag force. Microelectrodes of a suitable layout are used to trap one population of spheres, while the other one is dragged away from the electrodes by the generated fluid flow. The finite difference method is implemented in C++ to calculate the potential distribution by solving Laplace's equation. From the potential distribution, the DEP force on particles is calculated. The drag force on particles due to the liquid motion is calculated from the observed fluid velocity. The experimental results are shown to be in good agreement with the numerical solution. PMID- 16468934 TI - Aggregation profile characterisation in dielectrophoretic structures using bacteria and submicron latex particles. AB - A novel quantitative characterisation method for the measurement of anomalous low frequency aggregation processes on dielectrophoresis electrodes has been developed. Experimental evidence is provided for the relationship between the aggregation effect and AC electro-osmotical fluid motion theory. The aggregation profile dependence for E.coli bacteria, as a function of frequency and applied field, has been quantitatively examined. Additional experimental observations of the aggregation profiles of latex particles with dimensions of hundreds of nanometres, also confirm the relationship between this aggregation effect and the mentioned fluid motion theory. PMID- 16468935 TI - 3D focusing of nanoparticles in microfluidic channels. AB - Dynamic focusing of particles can be used to centre particles in a fluid stream, ensuring the passage of the particles through a specified detection volume. This paper describes a method for focusing nanoparticles using dielectrophoresis. The method differs from other focusing methods in that it manipulates the particles and not the fluid. Experimental focusing is demonstrated for a range of different particle types, and discussed in terms of the operational limits of the device. Dynamic numerical simulations of the particle motion in the device are presented and compared with the experimental results. The potential of the device for nanoparticle control and manipulation in microfluidic chips is discussed. PMID- 16468936 TI - Microdevices for separation, accumulation, and analysis of biological micro- and nanoparticles. AB - Microfabrication and performance of a novel microsystem for separation, accumulation and analysis of biological micro- and nanoparticles is reported. Versatile chip functions based on dielectrophoresis and microfluidics were integrated to isolate particles from complex sample solutions such as serum. A bead-based assay for virus detection is proposed. Separation of micro- and sub mum beads employing dielectrophoretic deflector and bandpass structures is demonstrated. Individual antibody coated beads with hepatitis A virus bound to their surface were trapped by negative dielectrophoresis in a field cage and analysed by fluorescence microscopy. PMID- 16468937 TI - Editorial: Molecular motors. PMID- 16468938 TI - Molecular motors: nature's nanomachines. AB - Molecular motors are protein-based machines that convert chemical potential energy into mechanical work. This paper aims to introduce the non-specialist reader to molecular motors, in particular, acto-myosin, the prototype system for motor protein studies. These motors produce their driving force from changes in chemical potential arising directly from chemical reactions and are responsible for muscle contraction and a variety of other cell motilities. PMID- 16468940 TI - Elucidation of the separate roles of myosins IIA and IIB during neurite outgrowth, adhesion and retraction. AB - The functions of nonmuscle myosin isoforms are key to an understanding of process outgrowth from nerve cells during animal development. Despite considerable structural similarity, myosin IIA and myosin IIB play distinct and complementary roles during the actin-based mechanisms of nerve process extension. An overview is given of evidence that implicates myosin IIB as the motor essential for nerve process outgrowth and myosin IIA both as the motor required to maintain cell adhesion to the substrate as well as the motor required to power retraction of the nerve cell process. These actions are placed in context within a model for nerve process extension that is consistent with many observations in the literature and provides testable hypotheses regarding possible roles for these nonmuscle myosin motors. The relevance of a fundamental understanding of the mechanisms underpinning nerve cell process extension to the application of nanotechnology in this area is also discussed. PMID- 16468939 TI - Modelling muscle motor conformations using low-angle X-ray diffraction. AB - New results on myosin head organization using analysis of low-angle X-ray diffraction patterns from relaxed insect flight muscle (IFM) from a giant waterbug, building on previous studies of myosin filaments in bony fish skeletal muscle (BFM), show that the information content of such low-angle diffraction patterns is very high despite the 'crystallographically low' resolution limit (65 A) of the spacings of the Bragg diffraction peaks being used. This high information content and high structural sensitivity arises because: (i) the atomic structures of the domains of the myosin head are known from protein crystallography; and (ii) myosin head action appears to consist mainly of pivoting between domains which themselves stay rather constant in structure, thus (iii) the intensity distribution among diffraction peaks in even the low resolution diffraction pattern is highly determined by the high-resolution distribution of atomically modelled domain mass. A single model was selected among 5000+ computer-generated variations as giving the best fit for the 65 reflections recorded within the selected resolution limit of 65 A. Clear evidence for a change in shape of the insect flight muscle myosin motor between the resting (probably like the pre-powerstroke) state and the rigor state (considered to mimic the end-of-powerstroke conformation) has been obtained. This illustrates the power of the low-angle X-ray diffraction method. The implications of these new results about myosin motor action during muscle contraction are discussed. PMID- 16468941 TI - Mathematical theory of molecular motors and a new approach for uncovering motor mechanism. AB - Molecular motors operate in an environment dominated by thermal fluctuations. A molecular motor may produce an active force at the reaction site to directly move the motor forward. Alternatively a molecular motor may generate a unidirectional motion by rectifying thermal fluctuations. In this case, the chemical reaction establishes free energy barriers to block the backward fluctuations. The effect of the chemical reaction on the motor motion can be represented by the motor potential profile (rectifying barrier andor active driving force). Different motor mechanisms are characterised by different motor potential profiles. The mathematical theory and properties of molecular motors are discussed and a mathematical framework is developed for extracting the motor potential profile from measured time series of motor position. As an example, we discuss the binding zipper model for the F(1) ATPase, which was motivated mainly by the fact that the motor potential profile of the F(1) ATPase is nearly a constant slope. PMID- 16468942 TI - Nanometre resolution tracking of myosin-1b motility. AB - The movement produced by a small number of myosin molecular motors was measured with nanometre precision using single-molecule fluorescence localisation methods. The positional precision of the measurements was sufficient to reveal fluctuations in sliding velocity due to stochastic interactions between individual myosin motors and the actin filament. Dependence of sliding velocity upon filament length was measured and fluctuations in velocity were quantified by autocorrelation analysis. Optical tweezers-based nanometry was used to measure the myosin-1b step-size directly. The 10 nm power-stroke and its duty cycle ratio were consistent with values derived from in vitro sliding assays. PMID- 16468943 TI - Polyunsaturated but not conjugated linoleic acid supplementation of leukemic U937 cells can act as an amplification factor for photofrin-mediated photodynamic therapy. AB - Polyunsaturated fatty acids located in leukemia cell membranes are excellent targets for peroxidation. They can significantly enhance the effectiveness of Photofrin-mediated photodynamic therapy (PDT)-induced cell killing. In this study, the peroxidizability of conjugated fatty acid isomers (9c,11t-linoleic acid and 9c,11c-linoleic acid) and polyunsaturated fatty acids (PUFAs; linoleic acid, gamma-linolenic acid and arachidonic acid) with 2,2'-azo-bis(2 amidinpropane)dihydrochloride, soybean lipoxygenase and photomediated peroxidation are compared with each other. Peroxidation was determined using different methods: by means of gas chromatography to estimate the fatty acid (FA) consumption, by photometry for the level of FA peroxides or phospholipid peroxides and by definition of the content of malondialdehyde for thiobarbituric acid reactive substances (TBARS). The results suggest that the generation of oxidation products from individual FAs indicate a different formation rate of oxidation products. Radical FA peroxides were produced most by polyunsaturated arachidonic acid, followed by linoleic acid and gamma-linolenic acid, whereas conjugated FA isomers did not generate peroxides. Accordingly, the levels of lipid peroxides and TBARS were substantially increased after incorporation and oxidation of polyunsaturated FAs into U937 cells and could significantly enhance the effectiveness of Photofrin-PDT-induced cytotoxicity. The results showed that PUFA, but not conjugated FA supplementation of U937 cells, can act as a PDT amplification factor. PMID- 16468945 TI - Behavioral assessment of intermittent wheel running and individual housing in mice in the laboratory. AB - Physical cage enrichment--exercise devices for rodents in the laboratory--often includes running wheels. This study compared responses of mice in enriched physical and social conditions and in standard social conditions to wheel running, individual housing, and open-field test. The study divided into 6 groups, 48 female BALB/c mice group housed in enriched and standard conditions. On alternate days, the study exposed 2 groups to individual running wheel cages. It intermittently separated from their cage mates and housed individually 2 groups with no running wheels; 2 control groups remained in enriched or standard condition cages. There were no significant differences between enriched and standard group housed mice in alternate days' wheel running. Over time, enriched, group housed mice ran less. Both groups responded similarly to individual housing. In open-field test, mice exposed to individual housing without running wheel moved more and faster than wheel running and home cage control mice. They have lower body weights than group housed and wheel running mice. Intermittent withdrawal of individual housing affects the animals more than other commodities. Wheel running normalizes some effects of intermittent separation from the enriched, social home cage. PMID- 16468944 TI - The career aspirations and location intentions of James Cook University's first cohort of medical students: a longitudinal study at course entry and graduation. AB - INTRODUCTION: James Cook University's School of Medicine enrolled its first cohort of 64 students in 2000. The School was established with the aim of increasing the number of medical graduates who understand rural, remote, Indigenous and tropical health issues and who would subsequently choose rural (non-metropolitan) practice. In December 2005, 58 students graduated the course. Several students left the cohort (either left the course or repeated earlier years of the course). In the early years, students who left the course were replaced by accelerated entry students. METHOD: A student-initiated survey, involving questionnaire and interview components, conducted in 2001 sought information on students' career aspirations and practice location intentions, as well as why students had chosen medicine and the James Cook University course in particular. A brief follow-up survey focused on career aspirations, location of internship and longer term practice location intentions was conducted in October 2005 (2 months before the students graduated). In each instance, participation was anonymous and no identifying information was recorded. Accelerated entry students' responses to the 2005 survey were included. RESULTS: In 2001, 57 (89%) of the 64 students participated in the study; and in 2005, 50 (86%) of the 58 graduating students responded to the questionnaire survey. Overall, the demographic composition of the 'cohort' changed little. In 2001, 66% of respondents intended to practice in non-metropolitan areas, with 64% so intending at graduation. This was reinforced by 64% choosing internship positions in non metropolitan areas of Queensland; 56% in north Queensland. Career aspirations changed appreciably between 2001 and 2005: the number of undecided students had halved, the numbers interested in general practice had reduced by one-third, the numbers considering surgery had reduced to one-third, and none was considering paediatrics at exit. Conversely, the number considering emergency medicine had almost doubled and more than doubled for obstetrics and gynaecology. DISCUSSION: Given that the majority of intern positions in Queensland are in the south-east of the state, it is noteworthy that two-thirds of students sought and accepted posts elsewhere, predominantly in northern Queensland. Changes in career aspirations reflect two issues. First, respondents' 2005 recollections of what their aspirations were in 2001 varied, reinforcing the value of the longitudinal process. Second, the change of intentions across time possibly reflects exposure to various disciplines and perhaps positive/negative experiences during clinical rotations in the final years of the course. The consistency across time with respect to intention to practice in rural (non-metropolitan) areas suggests that most respondents had eyes on rural practice from early in the course. The findings are in line with other studies that have demonstrated two important factors influence clinicians' decisions to take up non-metropolitan practice: (1) non-metropolitan background; and (ii) substantial and meaningful exposure to non metropolitan practice during the course. CONCLUSION: The findings support the School's contention and that of others around the world that medical education undertaken in non-metropolitan settings is the best vehicle for increasing the rural medical workforce. This study provides support for the development of regional medical schools that focus on local recruitment and health care need issues. PMID- 16468946 TI - Use of enclosure space by captive lion-tailed macaques (Macaca silenus) housed in Indian zoos. AB - Captive nonhuman animals use enclosure space differentially. Enclosure features strongly influence this. This study recorded both the enclosure space used by 47 captive lion-tailed macaques housed in 13 zoos across India and the behavior of the macaques. The exhibition of abnormal behaviors, food-related behaviors, and social interactions correlated significantly with the use of the edge zone (the part of the enclosure closest to the visitor area). Animals housed in barren enclosures used the edge zone to a significantly greater percentage than did those housed in complex exhibits. Percentages of autogrooming, social interactions, and food-related behaviors significantly correlated with the use of the enrich zone. Space use studies assist in recognizing areas within the enclosure, which captive animals actively use. Conversely, the studies can identify areas infrequently used and show how to make maximum use of these enclosure areas. Further studies targeting both the increase in percentages of natural behaviors exhibited and use of the enrich zone used the current study on captive lion-tailed macaques for their design. PMID- 16468948 TI - Caregiver perceptions of what indoor cats do "for fun". AB - In a survey conducted to determine enrichment activities for indoor cats in the caregivers' homes, 304 indoor-cat caregivers answered structured interviews about their cats' use of windows and other "fun activities." The survey asked caregivers about durations of window use, what cats watched through the windows, and in what other fun activities cats freely participated. The results indicate that, of 577 cats, 84.3% looked out windows a total of 5 hr or less a day, that cats observed 14 different categories of outdoor stimuli, and that cats participated in 17 categories of "other" fun activities. These results suggest that cats use windows and sunlight much less often than the welfare community may expect but that caregivers (owners), when possible, should consider providing access to these. PMID- 16468947 TI - Returning a recently adopted companion animal: adopters' reasons for and reactions to the failed adoption experience. AB - The return of a recently adopted companion animal places the nonhuman animal in jeopardy and may be painful and frustrating to the humans involved. However, if returners learn from the failed adoption experience, future adoptions may be more satisfactory for all concerned. In this study, 78 people who had adopted and returned dogs or cats to an animal shelter in a U.S. Midwestern city were interviewed regarding their reasons for return, reactions to the experience, and plans for future adoptions. Although some returners adjusted their pet ownership plans in potentially beneficial ways, most reacted by counseling greater forethought and planning before adopting. The last, although sound advice, had little to do with reasons for return, which primarily were problems that arose postadoption: pet behavior such as not getting along with other pets or children. Changing expectations about the development of new pet-family relationships and the provision of postadoption services might help adopters tolerate the adjustment period and handle problems without resorting to returning the animal. PMID- 16468949 TI - Welfare, health, and hygiene of laying hens housed in furnished cages and in alternative housing systems. AB - The aim of this review was to compare welfare, health, and hygienic status of laying hens housed in furnished cages and in alternative systems. In alternative systems (floor housing and aviaries), birds have more freedom of movement and a more complex environment than in furnished cages. However, housing birds in much larger flocks in alternative systems leads to an increased risk of feather pecking. Furthermore, air quality can be poorer in alternative systems than in furnished cages. This can affect health and hygienic status. There are only limited data on a direct comparison between furnished cages and alternative systems. Therefore, there is a need for an on-farm comparison of welfare, health, and hygienic status in these systems. PMID- 16468950 TI - Possible costs of radio-tracking a young adult female mantled howler monkey (Alouatta palliata) in deciduous habitat of Costa Rican tropical dry forest. PMID- 16468951 TI - Are we ready to give up on calcineurin inhibitors? PMID- 16468952 TI - C4d in liver allografts: a sign of antibody-mediated rejection? PMID- 16468953 TI - Management of hepatitis C in liver transplant recipients. AB - Recurrent hepatitis C virus (HCV) disease is the leading cause of graft loss in liver transplant recipients with pre-transplant HCV infection. While natural history is variable, median time to recurrent cirrhosis is less than a decade. Factors contributing to risk of recurrence and rate of fibrosis progression are only partially known. Older donor age, treatment of acute rejection, cytomegalovirus infection and high pre-transplant viral load are most consistently linked with worse outcomes. Whether these factors can be modified to positively impact on HCV disease progression is unknown. The main therapeutic approach for patients with recurrent HCV disease has been the treatment with interferon and ribavirin (RBV) once recurrent disease is documented or progressive. Efficacy is lower than in nontransplant patients and tolerability, especially of RBV, is a major limitation. Stable or improved fibrosis scores are seen in the majority of sustained responders. Optimal dose, duration and timing of treatment have not been determined. Alternative strategies under study include pre-transplant treatment of decompensated cirrhotics, preemptive antiviral therapy started within weeks of transplantation and prophylactic therapy using HCV antibodies. Ongoing studies may establish a future role for alternative treatment approaches. Additionally, limited overall efficacy of interferon-based therapy in the transplant setting highlights the urgent need for new drug therapies. PMID- 16468954 TI - Intravenous gammaglobulin (IVIG): a novel approach to improve transplant rates and outcomes in highly HLA-sensitized patients. AB - Intravenous immunoglobulin (IVIG) products are derived from pooled human plasma and have been used for the treatment of primary immunodeficiency disorders for more than 24 years. Shortly after their introduction, IVIG products were also found to be effective in the treatment of autoimmune and inflammatory disorders. Over the past 2 decades, the list of diseases where IVIG has a demonstrable beneficial effect has grown rapidly. These include Kawasaki disease, Guillain Barre syndrome, myasthenia gravis, dermatomyositis and demyelinating polyneuropathy. Recently, we have described a beneficial effect on the reduction of anti-HLA antibodies with subsequent improvement in transplantation of highly HLA-sensitized patients as well as a potent anti-inflammatory effect that is beneficial in the treatment of antibody-mediated rejection (AMR). These advancements have enabled transplantation of patients previously considered untransplantable. These studies and relevant mechanism(s) of action will be discussed here. PMID- 16468955 TI - Fat-derived hormone adiponectin combined with FTY720 significantly improves small for-size fatty liver graft survival. AB - Owing to the discrepancy between organ donation and the demand for liver transplantation, expanding the liver donor pool is of vital importance. However, marginal liver grafts, such as small-for-size and/or fatty grafts, were associated with primary graft nonfunction or poor function. Therefore, novel combination therapies to rescue small-for-size fatty liver grafts should be investigated. In this study, we applied a combination therapy using a fat-derived hormone adiponectin (anti-steatosis) plus immunomodulator FTY720 (anti inflammatory) in a rat liver transplantation model using small-for-size fatty liver grafts, and investigated the underlying protective mechanism such as anti steatosis, intra-graft energy metabolism, hepatic microcirculatory changes, cell signaling cascades for survival, apoptosis and inflammation. The current study demonstrated that even a single treatment of adiponectin or FTY720 improved the 7 day graft survival from 0% to 62.5% (p = 0.001). The combination therapy significantly increased the 7-day graft survival rate to 100% by remarkable attenuation of graft steatosis and acute phase inflammatory response, significant activation of cell survival Akt pathway and maintenance of intra-graft adenosine triphosphate metabolism and improvement of hepatic microcirculation. In conclusion, the fat-derived hormone adiponectin combined with FTY720 might be a novel combination drug therapy for prevention of small-for-size fatty liver graft injury. PMID- 16468957 TI - Link between immune cell infiltration and mitochondria-induced cardiomyocyte death during acute cardiac graft rejection. AB - Acute cardiac graft rejection (ACGR) is associated with cardiomyocyte apoptosis. We investigated the respective role of the Fas/FasL and mitochondrial permeability transition pore (mPTP) pathways in cardiomyocyte apoptosis accompanying ACGR. Heterotopic cardiac transplantations were performed in 7-9 week old C57BL6 or C3H mice. Wild type or Fas-deficient (lpr) mice underwent syngeneic (GS) or allogeneic (GA) transplantation, and received either saline or NIM811, a specific inhibitor of the mPTP. At day 5, we assessed ACGR by histology, cardiomyocyte apoptosis by caspase-3 activity and cytochrome c release, Ca(2+)-induced mPTP opening by a potentiometric approach, and expression of Fas, FasL, TNFalpha, perforin, granzyme using RT-PCR. Myocardial infiltration of CD8(+) T lymphocytes was performed by immunohistochemistry. Allogenic transplantation increased infiltration of inflammatory cells, upregulated FasL, perforin, granzyme, and TNFalpha, favored Ca(2+)-induced mPTP opening and increased caspase-3 activity and cytochrome c release in WT grafts. NIM811, but not Fas-deficiency, significantly reduced all these effects. NIM811 also limited infiltration of CD8(+) into WT and lpr transplants. These data suggest that the mPTP pathway plays a major role in cardiomyocyte apoptosis associated with ACGR. Inhibition of mPTP opening may attenuate cardiomyocyte apoptosis either directly or indirectly via a limitation of CD8(+) T-cell activation. PMID- 16468956 TI - Improved long-term graft survival after HO-1 induction in brain-dead donors. AB - Brain death (BD) of the donor, a risk factor uniquely relevant for organs derived from cadaver donors, influences organ quality by induction of various inflammatory events. Consequently ischemia/reperfusion injury is deteriorated and acute and chronic rejections accelerated. Donor treatment might be an approach to improve the quality of the graft. The induction of heme oxygenase 1 (HO-1) has been shown to exert beneficial effects in living-donor transplantation models. Therefore, we examined the impact of donor treatment with the selective inducer of HO-1, cobalt protoporphyrin (CoPP), on organ quality and transplant outcome in a standardized BD model in a F344-->LEW kidney transplant rat model. Immediately after BD induction, donor animals were administered a single dose of CoPP (5 mg/kg) and in control groups, HO-1 activity was blocked with zinc protoporphyrin (ZnPP, 20 mg/kg). Recipients of organs from brain-dead donors treated with CoPP survived significantly better than those from untreated brain-dead donors (p < 0.05) and intra-graft analysis showed improved histology (p < 0.05). Blockade of HO-1 with ZnPP decreased the survival rates (p < 0.05) comparable to untreated brain-dead donors. Our results demonstrate that HO-1 induction by one single treatment of CoPP in brain-dead donors leads to enhanced allograft survival. PMID- 16468958 TI - Modified two-layer preservation method (M-Kyoto/PFC) improves islet yields in islet isolation. AB - Islet allotransplantation can achieve insulin independence in patients with type I diabetes. Recent reports show that the two-layer method (TLM), which employs oxygenated perfluorochemical (PFC) and UW solution, is superior to simple cold storage in UW for pancreas preservation in islet transplantation. However, UW solution has several disadvantages, including the inhibition of Liberase activity. In this study, we investigated the features of a new solution, designated M-Kyoto solution. M-Kyoto solution contains trehalose and ulinastatin as distinct components. Trehalose has a cytoprotective effect against stress, and ulinastatin inhibits trypsin. In porcine islet isolation, islet yield was significantly higher in the M-Kyoto/PFC group compared with the UW/PFC group. There was no significant difference in ATP content in the pancreas between the two groups, suggesting that different islet yields are not due to their differences as energy sources. Compared with UW solution, M-Kyoto solution significantly inhibited trypsin activity in the digestion step; moreover, M-Kyoto solution inhibited collagenase digestion less than UW solution. In conclusion, the advantages of M-Kyoto solution are trypsin inhibition and less collagenase inhibition. Based on these data, we now use M-Kyoto solution for clinical islet transplantation from nonheart-beating donor pancreata. PMID- 16468959 TI - Role of natural killer cell subsets in cardiac allograft rejection. AB - To achieve donor-specific immune tolerance to allogeneic organ transplants, it is imperative to understand the cell types involved in acute allograft rejection. In wild-type mice, CD4(+) T cells are necessary and sufficient for acute rejection of cardiac allografts. However, when T-cell responses are suboptimal, such as in mice treated with costimulation-targeting agents or in CD28-deficient mice, and perhaps in transplanted patients taking immunosuppressive drugs, the participation of other lymphocytes such as CD8(+) T cells and NK1.1(+) cells becomes apparent. We found that host NK but not NKT cells were required for cardiac rejection. Ly49G2(+) NK cells suppressed rejection, whereas a subset of NK cells lacking inhibitory Ly49 receptors for donor MHC class I molecules was sufficient to promote rejection. Notably, rejection was independent of the activating receptors Ly49D and NKG2D. Finally, our experiments supported a mechanism by which NK cells promote expansion and effector function of alloreactive T cells. Thus, therapies aimed at specific subsets of NK cells may facilitate transplantation tolerance in settings of impaired T-cell function. PMID- 16468960 TI - Complete avoidance of calcineurin inhibitors in renal transplantation: a randomized trial comparing sirolimus and tacrolimus. AB - Calcineurin inhibitors have decreased acute rejection and improved early renal allograft survival, but their use has been implicated in the development of chronic nephrotoxicity. We performed a prospective, randomized trial in kidney transplantation comparing sirolimus-MMF-prednisone to tacrolimus-MMF-prednisone. Eighty-one patients in the sirolimus group and 84 patients in the tacrolimus group were enrolled (mean follow-up = 33 months; range 13-47 months). At 1 year, patient survival was similar in the groups (98% with sirolimus, 96% with tacrolimus; p = 0.42) as was graft survival (94% sirolimus vs. 92% tacrolimus, p = 0.95). The incidence of clinical acute rejection was 10% in the tacrolimus group and 13% in the sirolimus group (p = 0.58). There was no difference in mean GFR measured by iothalamate clearance between the tacrolimus and sirolimus groups at 1 year (61 +/- 19 mL/min vs. 63 +/- 18 mL/min, p = 0.57) or 2 years (61 +/- 17 mL/min vs. 61 +/- 19 mL/min, p = 0.84). At 1 year, chronicity using the Banff schema showed no difference in interstitial, tubular or glomerular changes, but fewer chronic vascular changes in the sirolimus group. This study shows that a CNI-free regimen using sirolimus-MMF-prednisone produces similar acute rejection rates, graft survival and renal function 1-2 years after transplantation compared to tacrolimus-MMF-prednisone. PMID- 16468961 TI - C4d in acute rejection after liver transplantation--a valuable tool in differential diagnosis to hepatitis C recurrence. AB - Hepatitis C is the most common indication for liver transplantation. Recurrence of HCV is universal leading to graft failure in up to 40% of all patients. The differentiation between acute rejection and recurrent hepatitis C is crucial as rejection treatments are likely to aggravate HCV recurrence. Histological examination of liver biopsy remains the gold standard for diagnosis of acute rejection but has failed in the past to distinguish between acute rejection and recurrent hepatitis C. We have recently reported that C4d as a marker of the activated complement cascade is detectable in hepatic specimen in acute rejection after liver transplantation. In this study, we investigate whether C4d may serve as a specific marker for differential diagnosis in hepatitis C reinfection cases. Immunohistochemical analysis of 97 patients was performed. A total of 67.7% of patients with acute cellular rejection displayed C4d-positive staining in liver biopsy whereas 11.8% of patients with hepatitis C reinfection tested positive for C4d. In the control group, 6.9% showed C4d positivity. For the first time we were able to clearly demonstrate that humoral components, represented by C4d deposition, play a role in acute cellular rejection after LTX. Consequently C4d may be helpful to distinguish between acute rejection and reinfection after LTX for HCV. PMID- 16468962 TI - Tacrolimus combined with two different dosages of sirolimus in kidney transplantation: results of a multicenter study. AB - Tacrolimus combined with mycophenolate mofetil (MMF) is an effective regimen in kidney transplantation. This study compared the efficacy of combining tacrolimus and two different dosages of sirolimus with an established tacrolimus-MMF regimen. Each day in addition to tacrolimus, 325 patients received 2 mg sirolimus (TAC-SRL2 mg), 325 patients received 0.5 mg sirolimus (TAC-SRL0.5 mg) and 327 patients 1 g MMF (TAC-MMF). The initial tacrolimus dose was 0.2 mg/kg/day. Sirolimus patients received loading doses of 6 or 1.5 mg, and daily doses of 2 or 0.5 mg thereafter. Steroid administration was identical for all groups. The incidence of biopsy-proven acute rejection was lower in the TAC-SRL2 mg group (15.7%) compared with the TAC-SRL0.5 mg (25.2%, p = 0.003) and the TAC-MMF groups (22.3%, p = 0.036). Six-month graft survival was 91.0% (TAC-SRL2 mg), 92.6% (TAC SRL0.5 mg) and 92.4% (TAC-MMF); the respective values for patient survival were 98.1%, 97.8% and 97.9%. Thirty-four patients (10.5%), 19 patients (5.8%) and 16 patients (4.9%) in the TAC-SRL2 mg, TAC-SRL0.5 mg and TAC-MMF groups, respectively, discontinued the study because of adverse events. Hyperlipemia was reported more often in the TAC-SRL2 mg group (24.0%) compared with 19.4% (TAC SRL0.5 mg) and 11.0% (TAC-MMF; p < 0.05). Combining 2 mg sirolimus/day with tacrolimus results in lower rates of acute rejection, but a higher incidence of adverse events. PMID- 16468963 TI - Explaining differences between hospitals in number of organ donors. AB - The shortage of donor organs calls for a careful examination of all improvement options. In this study, 80 Dutch hospitals were compared. They provided 868 donors in a 5-year period, constituting 91% of all donors in that period in The Netherlands. Multilevel regression analysis was used to explain the differences between hospitals. Potential explanatory variables were hospital-specific mortality statistics, donor policy and structural hospital characteristics. Of all donors, 81% came from one quarter of the hospitals, mainly larger hospitals. A strong relationship was found between the number of donors and hospital specific mortality statistics. Hospitals with a neurosurgery department had additional donors. Seven hospitals systematically underperformed over a period of 5 years. If these hospitals were to increase their donor efficiency to their expected value, it would lead to an increase of 10% in the number of donors. Most donors are found in large hospitals, implying that resources to improve donor recruitment should be channelled to larger hospitals. This study presents an efficient strategy toward a benchmark for hospitals of their organ donation rates. Some larger hospitals performed less well than others. This suggests that there is still room for improvement. There is no evidence for large undiscovered and unused pools of donor organs. PMID- 16468965 TI - Factors associated with publication following presentation at a transplantation meeting. AB - Full publication of abstracts presented at scientific meetings ranges from 25 74%. To determine the rate and factors associated with publication in organ transplantation, we examined abstracts presented at the American Transplant Congress in May 2000. Of 1147 abstracts, 607 (53%) achieved full publication at 4.5 years (mean 1.32 +/- 0.88 years). Fifty-nine percent (357/607) were published in three transplantation journals. For randomized trials, the proportion published was 61%. On multivariate analysis, industry sponsorship (OR 1.78; 95% CI 1.04-3.06), basic science research (OR 1.68; 95% CI 1.32-2.14), non-American center (OR 1.67; 95% CI 1.28-2.20) and oral presentation (OR 1.36; 95% CI 1.07 1.73) were independent predictors of full publication. Nearly half of all abstracts presented at a transplantation meeting remain unpublished. This finding needs to be considered when interpreting systematic reviews in the field of transplantation. PMID- 16468964 TI - Pre-implantation multiple cytokine mRNA expression analysis of donor lung grafts predicts survival after lung transplantation in humans. AB - While current donor selection with clinical findings is generally effective, the imprecise nature of the assessment forces clinicians to remain on the conservative side. A reliable biological marker would assist donor selection and would improve donor organ utilization. We collected biopsies from 169 donor lungs before implantation. Expression levels of IL-6, IL-8, IL-10, TNF-alpha, IFN-gamma and IL-1beta were measured by quantitative real-time RT-PCR (qRT-PCR). Seventeen cases died within 30 days after transplantation. No donor factor was significantly associated with 30-day mortality. Univariate analysis of the 84 cases for development of the prediction model showed that IL-6, IL-8, TNF-alpha and IL-1beta were risk factors for mortality and IL-10 and IFN-gamma were protective factors. We analyzed the cytokine expression ratios of risk to protective cytokines. A stepwise logistic regression for 30-day mortality demonstrated that a model containing the ratio of IL-6/IL-10 was the most predictive (p = 0.0013). When applied to the remaining 85 cases for validation, the test of model fit was significant (p = 0.039). Using the cytokine ratio, we were able to define three risk groups with striking differences in survival (p = 0.0003). Multi-cytokine analysis of the donor lung graft with qRT-PCR shows significant promise as a strategy to biologically evaluate the donor lung prior to implantation. PMID- 16468966 TI - Elective liver transplantation for the treatment of classical maple syrup urine disease. AB - An 8.5-year-old girl with classical maple syrup urine disease (MSUD) required liver transplantation for hypervitaminosis A and was effectively cured of MSUD over an 8-year clinical follow-up period. We developed a collaborative multidisciplinary effort to evaluate the effects of elective liver transplantation in 10 additional children (age range 1.9-20.5 years) with classical MSUD. Patients were transplanted with whole cadaveric livers under a protocol designed to optimize safe pre- and post-transplant management of MSUD. All patients are alive and well with normal allograft function after 106 months of follow-up in the index patient and a median follow-up period of 14 months (range 4-18 months) in the 10 remaining patients. Leucine, isoleucine and valine levels stabilized within 6 hours post-transplant and remained so on an unrestricted protein intake in all patients. Metabolic cure was documented as a sustained increase in weight-adjusted leucine tolerance, normalization of plasma concentration relationships among branched-chain and other essential and nonessential amino acids, and metabolic and clinical stability during protein loading and intercurrent illnesses. Costs and risks associated with surgery and immune suppression were similar to other pediatric liver transplant populations. PMID- 16468967 TI - Safety and immunogenicity of varicella-zoster virus vaccine in pediatric liver and intestine transplant recipients. AB - Primary varicella-zoster virus (VZV) infections following organ transplantation may cause significant morbidity. We examined the safety and immunogenicity of Varivax after transplantation as a potential prophylactic tool. Pediatric liver and intestine transplant recipients without history of chickenpox received one dose of Varivax. VZV humoral and cellular immunity were assessed before and > or =12 weeks after vaccination. Adverse events (AE) and management of exposure to wild type VZV were monitored. Sixteen VZV-naive subjects, 13-76 months of age, at 257-2045 days after transplantation were immunized. Five children developed mild local AE of short duration. Four subjects developed fever and four developed non injection site rashes, three of whom received acyclovir. Liver enzymes did not increase during the month after vaccination. Eighty-seven percent and 86% of children developed humoral and cellular immunity, respectively. There were five reported exposures to varicella in four children, none of which resulted in chickenpox. One subject received VZV-immunoglobulin and another subject with liver enzyme elevations after exposure received acyclovir; all remained asymptomatic. Varivax was safe and immunogenic in pediatric liver and intestine transplant recipients. Larger studies are needed to establish the efficacy and role of varicella vaccination after transplantation. PMID- 16468968 TI - Treatment of PTLD with rituximab or chemotherapy. AB - Information regarding treatment of post-transplant lymphoproliferative disease (PTLD) beyond reduction in immunosuppression (RI) is limited. We retrospectively evaluated patients receiving rituximab and/or chemotherapy for PTLD for response, time to treatment failure (TTF) and overall survival (OS). Thirty-five patients met inclusion criteria. Twenty-two underwent rituximab treatment, with overall response rate (ORR) 68%. Median TTF was not reached at 19 months and estimated OS was 31 months. In univariable analysis, Epstein-Barr virus (EBV) positivity predicted response and TTF. LDH elevation predicted shorter OS. No patient died of rituximab toxicity and all patients who progressed underwent further treatment with chemotherapy. Twenty-three patients received chemotherapy. ORR was 74%, median TTF was 10.5 months and estimated OS was 42 months. Prognostic factors for response included stage, LDH and allograft involvement by tumor. These factors and lack of complete response (CR) predicted poor survival. Twenty-six percent of the patients receiving chemotherapy died of toxicity. Rituximab and chemotherapy are effective in patients with PTLD who fail or do not tolerate RI. While rituximab is well tolerated, toxicity of chemotherapy is marked. PTLD patients requiring therapy beyond RI should be considered for rituximab, especially with EBV-positive disease. Chemotherapy should be reserved for patients who fail rituximab, have EBV-negative tumors or need a rapid response. PMID- 16468969 TI - CMV-specific CD8 T-cell dynamics in the blood and the lung allograft reflect viral reactivation following lung transplantation. AB - Despite the potentially high burden of cytomegalovirus (CMV)-related disease following lung transplantation, the role of the cytotoxic T-lymphocyte (CTL) response to CMV in this patient group is ill-defined. We assessed the CMV specific T-cell response in the blood and lung allograft of immunosuppressed lung transplant recipients receiving antiviral prophylaxis and following their withdrawal. While the proportion of CMV-specific CTL varied between patients, in the absence of CMV reactivation the level of CMV-specific CD8+ T cells in the blood remained stable over time. In the majority of patients CMV-specific cells could be detected in the lung allograft, often in the absence of viral DNA. Additionally, following primary CMV lung infection, CMV-specific CD8+ T cells were detected no earlier than 100 days post-transplantation but still prior to the detection of viral DNA in the lung allograft. Together these findings suggest that very low levels of CMV replication are sufficient to both prime and recruit CMV-specific CD8+ T cells to the MHC-mismatched lung allograft. The direct detection of CMV-specific T cells with an effector phenotype in the lung allograft suggests a protective antiviral function. This study provides a framework upon which the association between CMV and chronic allograft rejection can be further studied. PMID- 16468971 TI - Improved survival after live donor adult liver transplantation (LDALT) using right lobe grafts: program experience and lessons learned. AB - We present our program experience with 85 live donor adult liver transplantation (LDALT) procedures using right lobe grafts with five simultaneous live donor kidney transplants using different donors performed over a 6-year period. After an "early" 2-year experience of 25 LDALT procedures, program improvements in donor and recipient selection, preoperative imaging, donor and recipient surgical technique and immunosuppressive management significantly reduced operative mortality (16% vs. 3.3%, p = 0.038) and improved patient and graft 1-year survival in recipients during our "later" experience with the next 60 cases (January 2001 and March 2005; patient survival: early 70.8% vs. later 92.7%, p = 0.028; graft survival: Early 64% vs. later 91.1%, p = 0.019, respectively). Overall patient and graft survival were 82% and 80%. There was a trend for less postoperative complications (major and minor) with program experience (early 88% vs. later 66.7%; p = 0.054) but overall morbidity remained at 73.8%. Biliary complications (cholangitis, disruption, leak or stricture) were not influenced by program experience (early 32% vs. later 38%). Liver volume adjusted to 100% of standard liver volume (SLV) within 1 month post-transplant. Despite a high rate of morbidity after LDALT, excellent patient and graft survival can be achieved with program experience. PMID- 16468970 TI - Decreased risk of renal allograft thrombosis associated with interleukin-2 receptor antagonists: a report of the NAPRTCS. AB - Graft thrombosis is the most common cause of first year graft failure in pediatric renal transplantation. The North American Pediatric Renal Transplant Cooperative Study (NAPRTCS) database was analyzed for cases of graft failure due to thrombosis among patients transplanted from 1998 to 2004. The impact of interleukin-2 (IL-2) receptor antagonists as induction therapy was determined. There were a total of 51 graft failures due to thrombosis among the 2750 reported renal transplants (1.85%) (95% CI (1.39%, 2.41%)). This represents the most common cause of graft loss during the first year post-transplant accounting for 35% of first year losses and 18% of all graft losses. The incidence of thrombosis among patients who received IL-2 receptor antibodies was 1.07% (12/1126) compared to 2.40% (39/1624) among patients who did not (OR 0.44, 95% CI 0.23, 0.84, p = 0.014). Use of IL-2 receptor blockade was the only significant prognostic factor in a multivariate model with previously identified risk factors. Analysis of NAPRTCS data found that the use of IL-2 receptor antibodies as induction therapy is associated with a significantly decreased risk of graft failure due to thrombosis. This provocative finding requires further investigation to determine whether thrombotic failure can be decreased by this therapeutic strategy. PMID- 16468972 TI - The dynamics and associations of airway neutrophilia post lung transplantation. AB - Bronchoalveolar lavage (BAL) neutrophilia has been repeatedly observed in lung transplant recipients with established bronchiolitis obliterans syndrome (BOS). Little is known of the fluctuations in BAL and airway neutrophilic inflammation post-transplant. This prospective longitudinal study aimed to evaluate the dynamic changes of lung allograft neutrophils with time, immunosuppression, infection and BOS. A total of 28, initially healthy, BOS 0, lung transplant recipients underwent 134 bronchoscopic assessments, including BAL and endobronchial biopsies (EBB) (with immunohistochemistry) over 3-year follow up. Subsequently, 21 developed BOS 0p and 16 ultimately BOS. Compared to controls, there was early and persistent BAL neutrophilia (p < 0.05), contrasting with an initially normal EBB that shows a progressive increased airway wall neutrophil infiltrate. BAL neutrophilia (but not airway wall neutrophilia) was most striking when there was concomitant bronchopulmonary infection, particularly in the patients with BOS. Univariate and multivariate analyses suggested that BAL neutrophilia was linked to markers of infection while EBB neutrophilia was linked with coexistent inflammation with macrophages and lymphocytes. IN CONCLUSION: (i) BAL neutrophilia is predominantly associated with infection; (ii) Airway wall neutrophilia (as monitored by EBB) increases with time post-transplant and is not associated with infection; (iii) By itself, BOS is not the major contributor to BAL and EBB neutrophilia. PMID- 16468973 TI - Real-time contrast-enhanced sonography of renal transplant recipients predicts chronic allograft nephropathy. AB - Real-time contrast-enhanced sonography (RT-CES) can assess microvascular tissue perfusion using gas-filled microbubbles. The study was performed to evaluate the feasibility of RT-CES in detecting chronic allograft nephropathy (CAN) in comparison to color Doppler ultrasonography (CDUS). A total of 26 consecutive renal transplant recipients were prospectively studied using RT-CES and conventional CDUS. Transplant tissue perfusion imaging was performed by low-power imaging during i.v. administration of the sonocontrast Optison. Renal tissue perfusion was assessed quantitatively using flash replenishment kinetics of microbubbles to estimate renal blood flow A *beta (A = peak signal intensity, beta= slope of signal intensity rise). In contrast to conventional CDUS resistance and pulsatility indices, renal blood flow estimated by CES was highly significant related to S-creatinine (r =-0.62, p = 0.0004). Determination of renal blood flow by CES reached a higher sensitivity (91% vs. 82%, p < 0.05), specificity (82% vs. 64%, p < 0.05) and accuracy (85% vs. 73%, p < 0.05) for the diagnosis of CAN as compared to conventional CDUS resistance indices. Perfusion parameters derived from RT-CES significantly improve the early detection of CAN compared to conventional CDUS. RT-CES using low-power real-time perfusion imaging is a feasible method to evaluate microvascular perfusion in renal allograft recipients. PMID- 16468974 TI - Donor kidney volume and outcomes following live donor kidney transplantation. AB - Pre-donation kidney volume and function may be crucial factors in determining graft outcomes in kidney transplant recipients. We measured living donor kidney volumes by 3D helical computed tomography scanning and glomerular filtration rate (GFR) by (125)I-iothalamate clearances in 119 donors, and correlated these values with graft function and incidence of acute rejection at 2 years post transplantation. Kidney volume strongly correlated with GFR (Pearson r= 0.71, p < 0.001). Body size and male gender were independent correlates of larger kidney volumes, and body size and age were predictors of kidney function. The effects of transplanted kidney volume on graft outcome were studied in 104 donor-recipient pairs. A transplanted kidney volume greater than 120 cc/1.73 m(2) was independently associated with better estimated GFR at 2 years post-transplant when compared to recipients of lower transplanted kidney volumes (64 +/- 19 vs. 48 +/- 14 mL/min/1.73 m(2), p < 0.001). Moreover, recipients of lower volumes had a higher incidence of acute cellular rejection (16% vs. 3.7%, p = 0.046). In conclusion, kidney volume strongly correlates with function in living kidney donors and is an independent determinant of post-transplant graft outcome. The findings suggest that (1) transplantation of larger kidneys confers an outcome advantage and (2) larger kidneys should be preferred when selecting from otherwise similar living donors. PMID- 16468976 TI - Successful hepatorenal transplantation in hereditary amyloidosis caused by a frame-shift mutation in fibrinogen Aalpha-chain gene. AB - Hereditary systemic amyloidosis comprises several autosomal dominant diseases caused by mutations in a number of plasma proteins, including the fibrinogen Aalpha-chain. Four mutations in the fibrinogen Aalpha-chain that are able to induce amyloidosis have been identified so far, the most common being the Glu526Val mutation. We have observed a family in which the father and his son reached end-stage renal failure because of renal amyloidosis induced by a frame shift mutation in the fibrinogen Aalpha-chain gene producing a novel amyloid protein. Two kidney transplantations in the father and one in the son resulted in fast graft loss caused by recurrence of amyloid deposition. We then performed hepatorenal transplantation in the son. Three years later, liver and kidney functions are normal without recurrence of amyloid deposition. This case, together with three others with the Glu526Val mutation in the extensive literature, suggests that liver transplantation can cure hereditary fibrinogen amyloidosis, whatever the mutation may be. PMID- 16468975 TI - HLA type-independent method to monitor polyoma BK virus-specific CD4 and CD8 T cell immunity. AB - (Re)activation of quiescent viral diseases is a major problem in immunosuppressed transplant patients. Polyoma BK virus-associated nephropathy (PVAN) caused by active polyoma BK virus (BKV) infection became a main reason for graft loss in kidney transplantation. After diagnosis, most transplant centers react by reducing immunosuppression (IS) to allow the immune system to control the infection. However, the impact of reduced IS on BKV immunity is not well researched. Here we present an HLA type-independent method to monitor BKV specific T-cell immunity. Applying our method, viral protein 1-specific CD4+ and CD8+ T-cell responses were detected in patients with serum BKV-DNA levels >250 000 copies/mL. In addition, specific T-cell responses were also found in allograft-infiltrating cells. The method can be used to assess the impact of decreased immunosuppression on BKV immunity and to clarify the role of specific T cells in the pathogenesis of PVAN. We strongly recommend its implementation in future clinical studies. PMID- 16468977 TI - Successful renal transplantation in patients with chronic granulomatous disease. AB - Chronic granulomatous disease (CGD) is a genetic disease caused by structural mutations in the enzyme NADPH oxidase that results in severe immunodeficiency. End-stage renal disease occurs in this patient population, and is often attributed to the necessary use of nephrotoxic anti-infectives. In this report, we present the experiences of two centers in transplantation of three patients with CGD: one transplanted with CGD, one cured of his CGD with bone marrow transplantation who subsequently underwent kidney transplantation and one that received a kidney transplant prior to being cured of CGD via a sequential peripheral blood stem cell transplant (SCT). All three recipients have enjoyed excellent outcomes. Their courses demonstrate the absolute requirements for a multidisciplinary and compulsive approach before, during and after transplantation. These case reports also highlight the unexpectedly benign effects of immunosuppressive therapy in this patient population. PMID- 16468978 TI - HLA susceptibility to BKV infection. PMID- 16468980 TI - The ingredients of a successful donation by brain death program in Greece. PMID- 16468981 TI - Chromosome healing by de novo telomere addition in Saccharomyces cerevisiae. AB - The repair of spontaneous or induced DNA damage by homologous recombination (HR) in Saccharomyces cerevisiae will suppress chromosome rearrangements. Alternative chromosome healing pathways can result in chromosomal instability. One of these pathways is de novo telomere addition where the end of a broken chromosome is stabilized by telomerase-dependent addition of telomeres at non-telomeric sites. De novo telomere addition requires the recruitment of telomerase to chromosomal targets. Subsequently, annealing of the telomerase reverse transcriptase RNA template (guide RNA) at short regions of homology is followed by extension of the nascent 3'-end of the broken chromosome to copy a short region of the telomerase guide RNA; multiple cycles of this process yield the new telomere. Proteins including Pif1 helicase, the single-stranded DNA-binding protein Cdc13 and the Ku heterocomplex are known to participate in native telomere functions and also regulate the de novo telomere addition reaction. Studies of the sequences added at de novo telomeres have lead to a detailed description of the annealing extension-dissociation cycles that copy the telomerase guide RNA, which can explain the heterogeneity of telomeric repeats at de novo and native telomeres in S. cerevisiae. PMID- 16468982 TI - CelTOS, a novel malarial protein that mediates transmission to mosquito and vertebrate hosts. AB - The malarial parasite has two hosts in its life cycle, a vertebrate and a mosquito. We report here that malarial invasion into these hosts is mediated by a protein, designated cell-traversal protein for ookinetes and sporozoites (CelTOS), which is localized to micronemes that are organelles for parasite invasive motility. Targeted disruption of the CelTOS gene in Plasmodium berghei reduced parasite infectivity in the mosquito host approximately 200-fold. The disruption also reduced the sporozoite infectivity in the liver and almost abolished its cell-passage ability. Liver infectivity was restored in Kupffer cell-depleted rats, indicating that CelTOS is necessary for sporozoite passage from the circulatory system to hepatocytes through the liver sinusoidal cell layer. Electron microscopic analysis revealed that celtos-disrupted ookinetes invade the midgut epithelial cell by rupturing the cell membrane, but then fail to cross the cell, indicating that CelTOS is necessary for migration through the cytoplasm. These results suggest that conserved cell-passage mechanisms are used by both sporozoites and ookinetes to breach host cellular barriers. Elucidation of these mechanisms might lead to novel antimalarial strategies to block parasite's transmission. PMID- 16468983 TI - Aspergillus nidulans class V and VI chitin synthases CsmA and CsmB, each with a myosin motor-like domain, perform compensatory functions that are essential for hyphal tip growth. AB - The polarized synthesis of cell wall components such as chitin is essential for the hyphal tip growth of filamentous fungi. The actin cytoskeleton is known to play important roles in the determination of hyphal polarity in Aspergillus nidulans. Previously, we suggested that CsmA, a chitin synthase with a myosin motor-like domain (MMD), was involved in polarized chitin synthesis in a manner dependent on the interaction between the MMD and the actin cytoskeleton. The genome database indicates that A. nidulans possesses another gene encoding another chitin synthase with an MMD. In this study, we characterized this gene, which we designated csmB. The csmB null mutants examined were viable, although they exhibited defective phenotypes, including the formation of balloons and intrahyphal hyphae and the lysis of subapical regions, which were similar to those obtained with csmA null mutants. Moreover, csmA csmB double null mutants were not viable. Mutants in which csmB was deleted and the expression of csmA was under the control of the alcA promoter were viable but severely impaired in terms of hyphal growth under alcA-repressing conditions. We revealed that CsmB with three copies of a FLAG epitope tag localized at the hyphal tips and forming septa, and that the MMD of CsmB was able to bind to actin filaments in vitro. These results suggest that CsmA and CsmB perform compensatory functions that are essential for hyphal tip growth. PMID- 16468984 TI - The role of proton motive force in expression of the Staphylococcus aureus cid and lrg operons. AB - The Staphylococcus aureus cidABC and lrgAB operons have been shown to play a key role in the regulation of murein hydrolase activity and cell death in a manner thought to be analogous to bacteriophage-encoded holins and anti-holins respectively. Because of these functions, it has been proposed that the regulation of these operons is tightly controlled and responsive to key metabolic signals. The current study revealed the presence of two overlapping regulatory pathways controlling cidABC and lrgAB expression, one dependent on acetic acid and the other dependent on proton motive force (PMF). The latter pathway was analysed using agents that affect various aspects of the PMF. Gramicidin and carbonyl cyanide m-chlorophenylhydrazone (CCCP), antimicrobial agents that dissipate the DeltapH and membrane potential (DeltaPsi), both enhanced lrgAB expression. Restoration of the PMF by incubation of the bacteria in the presence of glucose restored lrgAB expression back to the uninduced state. In addition, valinomycin, which specifically collapses the DeltaPsi, also induced lrgAB expression. In contrast, nigericin, which dissipates the DeltapH component of the PMF, was found to have a minimal effect on DeltaPsi and lrgAB transcription. Finally, the DeltaPsi-inducible expression of lrgAB was shown to be dependent on the previously characterized LytSR two-component regulatory system that is involved in the regulation of autolysis. The results of this study support a model in which the LytSR regulatory system responds to a collapse in DeltaPsi by inducing the transcription of the lrgAB operon. PMID- 16468985 TI - Mycobacterium tuberculosis prcBA genes encode a gated proteasome with broad oligopeptide specificity. AB - Genes predicted to be associated with the putative proteasome of Mycobacterium tuberculosis (Mtb) play a critical role in defence of the bacillus against nitrosative stress. However, proteasomes are uncommon in eubacteria and it remains to be established whether Mtb's prcBA genes in fact encode a proteasome. We found that coexpression of recombinant PrcB and PrcA in Escherichia coli over a prolonged period at 37 degrees C allowed formation of an alpha(7)beta(7)beta(7)alpha(7), 750 kDa cylindrical stack of four rings in which all 14 beta-subunits were proteolytically processed to expose the active site threonine. In contrast to another Actinomycete, Rhodococcus erythropolis, Mtb's beta-chain propeptide was not required for particle assembly. Peptidolytic activity of the 750 kDa particle towards a hydrophobic oligopeptide was nearly two orders of magnitude less than that of the Rhodococcus 20S proteasome, and unlike eukaryotic and archaeal proteasomes, activity of the Mtb 750 kDa particle could not be stimulated by SDS, Mg(2+) or Ca(2+). Electron microscopy revealed what appeared to be obstructed alpha-rings in the Mtb 750 kDa particle. Deletion of the N-terminal octapeptide from Mtb's alpha-chain led to disappearance of the apparent obstruction and a marked increase of peptidolytic activity. Unlike proteasomes isolated from other Actinomycetes, the open-gate Mtb mutant 750 kDa particle cleaved oligopeptides not only after hydrophobic residues but also after basic, acidic and small, neutral amino acids. Thus, Mtb encodes a broadly active, gated proteasome that may work in concert with an endogenous activator. PMID- 16468986 TI - Structure of the Mycobacterium tuberculosis proteasome and mechanism of inhibition by a peptidyl boronate. AB - Mycobacterium tuberculosis (Mtb) has the remarkable ability to resist killing by human macrophages. The 750 kDa proteasome, not available in most eubacteria except Actinomycetes, appears to contribute to Mtb's resistance. The crystal structure of the Mtb proteasome at 3.0 A resolution reveals a substrate-binding pocket with composite features of the distinct beta1, beta2 and beta5 substrate binding sites of eukaryotic proteasomes, accounting for the broad specificity of the Mtb proteasome towards oligopeptides described in the companion article [Lin et al. (2006), Mol Microbiol doi:10.1111/j.1365-2958.2005.05035.x]. The substrate entrance at the end of the cylindrical proteasome appears open in the crystal structure due to partial disorder of the alpha-subunit N-terminal residues. However, cryo-electron microscopy of the core particle reveals a closed end, compatible with the density observed in negative-staining electron microscopy that depended on the presence of the N-terminal octapetides of the alpha-subunits in the companion article, suggesting that the Mtb proteasome has a gated structure. We determine for the first time the proteasomal inhibition mechanism of the dipeptidyl boronate N-(4-morpholine)carbonyl-beta-(1-naphthyl)-L-alanine-L leucine boronic acid (MLN-273), an analogue of the antimyeloma drug bortezomib. The structure improves prospects for designing Mtb-specific proteasomal inhibitors as a novel approach to chemotherapy of tuberculosis. PMID- 16468987 TI - CRZ1, a target of the calcineurin pathway in Candida albicans. AB - Calcineurin is a major player in calcium-dependent signal transduction pathways of eukaryotes. Calcineurin acts on transcription factors (e.g. CRZ1 in Saccharomyces cerevisiae) and governs the expression of genes in a species dependent fashion. In Candida albicans, the calcineurin pathway is involved in tolerance to antifungal agents, cation homeostasis and virulence. However, the components of the calcineurin pathway are still poorly investigated in this yeast species. Taking S. cerevisiae as a model to reconstitute this pathway, two CRZ1 like genes, CRZ1 and CRZ2 (for calcineurin-responsive zinc finger 1 and 2 genes), were found with C(2)H(2) zinc finger domains. Only CRZ1 was able to restore the calcium hypersusceptibility of a S. cerevisiae crz1Delta mutant and to mediate calcium-dependent gene expression in this yeast species. Several experiments showed that CRZ1 was dependent on calcineurin in C. albicans: (i) phenotypic analysis of a crz1Delta/Delta mutant showed impaired growth as compared with the wild type in the presence of cations (Ca(2+), Mn(2+)) as does a mutant lacking calcineurin subunit A (cnaDelta/Delta) and (ii) a green fluorescent protein (GFP) Crz1p fusion protein showed a calcium- and calcineurin-dependent nuclear localization. To further analyse the relationship between calcineurin and CRZ1, a comprehensive analysis of calcineurin/Crz1p-dependent gene expression following addition of Ca(2+) (200 mM) was performed. Among the expression of 264 genes altered by at least twofold, the upregulation of 60 genes was dependent on both calcineurin and CRZ1. Interestingly, a motif [5'-G(C/T)GGT-3'] with similarity to the target sequence of Crz1p (GNGGCG/TCA) from S. cerevisiae was identified as a putative regulatory sequence in the upstream regions of these calcineurin/Crz1p dependent genes. However, additional experiments showed that calcineurin may have other targets in addition to CRZ1. First, CRZ1 was not involved in tolerance to antifungal agents (fluconazole, terbinafine) on the opposite to calcineurin. Second, CRZ1 was only moderately influencing virulence in a mice model of infection which is in sharp contrast to the strong avirulence of cnaDelta/Delta mutant in the same animal model. Even though this work establishes CRZ1 as a calcineurin target, further studies are needed to identify other calcineurin dependent elements in C. albicans. PMID- 16468989 TI - Borrelia burgdorferi lipoproteins are secreted to the outer surface by default. AB - Borrelia spirochaetes are unique among diderm bacteria in their abundance of surface-displayed lipoproteins, some of which play important roles in the pathogenesis of Lyme disease and relapsing fever. To identify the lipoprotein sorting signals in Borrelia burgdorferi, we generated chimeras between the outer surface lipoprotein OspA, the periplasmic oligopeptide-binding lipoprotein OppAIV and mRFP1, a monomeric red fluorescent reporter protein. Localization of OspA and OppAIV point mutants showed that Borrelia lipoproteins do not follow the '+2' sorting rule which targets lipoproteins to the cytoplasmic or outer membrane of Gram-negative bacteria via the Lol pathway. Fusions of mRFP1 to short N-terminal lipopeptides of OspA, and surprisingly OppAIV, were targeted to the spirochaetal surface. Mutagenesis of the OspA N-terminus defined less than five N-terminal amino acids as the minimal secretion-facilitating signal. With the exception of negative charges, which can act as partial subsurface retention signals in certain peptide contexts, lipoprotein secretion occurs independent of N-terminal sequence. Together, these data indicate that Borrelia lipoproteins are targeted to the bacterial surface by default, but can be retained in the periplasm by sequence-specific signals. PMID- 16468988 TI - Rad52 depletion in Candida albicans triggers both the DNA-damage checkpoint and filamentation accompanied by but independent of expression of hypha-specific genes. AB - We have analysed the effect of RAD52 deletion in several aspects of the cell biology of Candida albicans. Cultures of rad52Delta strains exhibited slow growth and contained abundant cells with a filamentous morphology. Filamentation with polarization of actin patches was accompanied by the induction of the hypha specific genes (HSG) ECE1, HWP1 and HGC1. However, filament formation occurred in the absence of the transcription factors Efg1 and Cph1, even though disruption of EFG1 prevented expression of HSG. Therefore, expression of HSG genes accompanies but is dispensable for rad52Delta filamentation. However, deletion of adenylate cyclase severely impaired filamentation, this effect being largely reverted by the addition of exogenous cAMP. Filaments resembled elongated pseudohyphae, but some of them looked like true hyphae. Following depletion of Rad52, many cells arrested at the G2/M phase of the cell cycle with a single nucleus suggesting the early induction of the DNA-damage checkpoint. Filaments formed later, preferentially from G2/M cells. The filamentation process was accompanied by the uncoupling of several landmark events of the cell cycle and was partially dependent on the action of the cell cycle modulator Swe1. Hyphae were still induced by serum, but a large number of rad52 cells myceliated in G2/M. PMID- 16468991 TI - Replication-associated gene dosage effects shape the genomes of fast-growing bacteria but only for transcription and translation genes. AB - The bidirectional replication of bacterial genomes leads to transient gene dosage effects. Here, we show that such effects shape the chromosome organisation of fast-growing bacteria and that they correlate strongly with maximal growth rate. Surprisingly the predicted maximal number of replication rounds shows little if any phylogenetic inertia, suggesting that it is a very labile trait. Yet, a combination of theoretical and statistical analyses predicts that dozens of replication forks may be simultaneously present in the cells of certain species. This suggests a strikingly efficient management of the replication apparatus, of replication fork arrests and of chromosome segregation in such cells. Gene dosage effects strongly constrain the position of genes involved in translation and transcription, but not other highly expressed genes. The relative proximity of the former genes to the origin of replication follows the regulatory dependencies observed under exponential growth, as the bias is stronger for RNA polymerase, then rDNA, then ribosomal proteins and tDNA. Within tDNAs we find that only the positions of the previously proposed 'ubiquitous' tRNA, which translate the most frequent codons in highly expressed genes, show strong signs of selection for gene dosage effects. Finally, we provide evidence for selection acting upon genome organisation to take advantage of gene dosage effects by identifying a positive correlation between genome stability and the number of simultaneous replication rounds. We also show that gene dosage effects can explain the over representation of highly expressed genes in the largest replichore of genomes containing more than one chromosome. Together, these results demonstrate that replication-associated gene dosage is an important determinant of chromosome organisation and dynamics, especially among fast-growing bacteria. PMID- 16468990 TI - Ammonium permease-based sensing mechanism for rapid ammonium activation of the protein kinase A pathway in yeast. AB - In the yeast Saccharomyces cerevisiae starvation for nitrogen on a glucose containing medium causes entrance into G0 and downregulation of all targets of the PKA pathway. Re-addition of a nitrogen source in the presence of glucose causes rapid activation of trehalase and other PKA targets. Trehalase activation upon ammonium re-supplementation is dependent on PKA activity, but not on its regulatory subunit nor is it associated with an increase in cAMP. In nitrogen starved cells, ammonium transport and activation of trehalase are most active in strains expressing either the Mep2 or Mep1 ammonium permease, as opposed to Mep3. The non-metabolizable ammonium analogue, methylamine, also triggers activation of trehalase when transported by Mep2 but not when taken up by diffusion. Inhibition of ammonium incorporation into metabolism did not prevent signalling. Extensive site-directed mutagenesis of Mep2 showed that transport and signalling were generally affected in a similar way, although they could be separated partially by specific mutations. Our results suggest an ammonium permease-based sensing mechanism for rapid activation of the PKA pathway. Mutagenesis of Asn246 to Ala in Mep2 abolished transport and signalling with methylamine but had no effect with ammonium. The plant AtAmt1;1, AtAmt1;2, AtAmt1;3 and AtAmt2 ammonium transporters sustained transport and trehalase activation to different extents. Specific mutations in Mep2 affected the activation of trehalase differently from induction of pseudohyphal differentiation. We also show that Mep permease involvement in PKA control is different from their role in haploid invasive growth, in which Mep1 sustains and Mep2 inhibits, in a way independent of the ammonium level in the medium. PMID- 16468992 TI - A slipped-mispairing mutation in AgrA of laboratory strains and clinical isolates results in delayed activation of agr and failure to translate delta- and alpha haemolysins. AB - agr is a global regulator of staphylococcal virulence and other accessory gene functions, especially including the haemolysins. Lack of haemolysin production therefore generally represents a defect in agr function. An example of this is Staphylococcus aureus strain RN4220, a widely used laboratory strain that carries a nitrosoguanidine (MNNG)-induced mutation enabling it to accept DNA from Escherichia coli and other bacteria. We show here that the non-haemolytic phenotype of RN4220 is caused by an extra A residue in a run of seven As at the 3' end of agrA (agrA-8A). This causes a frameshift that results in the addition of three amino acyl residues to the C-terminal end of the protein. The 8A mutation does not inactivate the agr locus, but rather delays agr activation by 2 3 h, which results in failure to translate alpha- and delta-haemolysins, and hence, in a non-haemolytic phenotype. This mutation turned out not to be an adventitious consequence of MNNG mutagenesis, but rather had arisen in RN450, the immediate parent of RN4220. RN450 had become haemolytically heterogeneous in storage, and its non-haemolytic variants had the 8A mutation. The same mutation was also identified in a clinical isolate in which a non-haemolytic variant had arisen during the course of infection. Haemolytic activity in the mutant laboratory strains could be restored by the addition of auto-inducing peptide (AIP) early in growth, indicating that delayed production of RNAIII is responsible for the failure to translate alpha- and delta-haemolysins. Discovery of the 8A mutation has revealed the basis of the dissociation between agr activity and the non-haemolytic phenotype of RN4220, and has solved the long standing mystery of the variable non-haemolytic phenotype of its immediate parent, RN450. The occurrence of this mutation in a clinical isolate indicates that it is not simply a laboratory phenomenon, and may represent a naturally occurring mechanism for the modulation of agr activity. PMID- 16468993 TI - The ISWI and CHD1 chromatin remodelling activities influence ADH2 expression and chromatin organization. AB - Nucleosome remodelling complexes play a key role in gene activation in response to environmental changes by driving promoter chromatin to reach an accessible configuration. They also mediate genome-wide chromatin organization, although their role in processes other than activation-related chromatin remodelling are poorly understood. The Saccharomyces cerevisiae ADH2 gene represents an excellent model for understanding the role of chromatin structure and remodelling in gene regulation. Following glucose depletion, highly positioned promoter nucleosomes are destabilized leading to strictly regulated kinetics of transcriptional activation. Nevertheless, no chromatin remodelling activities responsible for establishing or remodelling ADH2 chromatin structure have been identified to date. Here we show that the absence of the Isw1 and Chd1 ATP-dependent chromatin remodelling activities delays the maximal expression of ADH2 without impairing the chromatin remodelling that occurs upon activation. Instead, a destabilized chromatin structure on the ADH2 coding and termination region is observed in the absence of Isw1 or Chd1 in repressing conditions. The specific Isw1 complex involved in this nucleosome repositioning is Isw1b because the deletion of Ioc2 and Ioc4, but not of Ioc3, causes the same phenotype as the deletion of Isw1. Moreover, the lack of Chd1 combined with the absence of Isw1 and Isw2 impairs nucleosome spacing along the ADH2 gene, and genome-wide in S. cerevisiae. Thus, the ISWI and Chd1 remodelling factors are not only involved in transcription related chromatin remodelling, but also are required to maintain a specific chromatin configuration across the yeast genome. PMID- 16468994 TI - Hfq-dependent alterations of the transcriptome profile and effects on quorum sensing in Pseudomonas aeruginosa. AB - The Pseudomonas aeruginosa quorum-sensing (QS) systems, Las and Rhl, control the production of several virulence factors and other proteins, which are important to sustain adverse conditions. A comparative transcriptome analysis of a rpoS (-) and a rpoS(-)hfq( -) strain indicated that the Sm-like RNA-binding protein Hfq affects approximately 5% of the P. aeruginosa O1 transcripts. Among these transcripts 72 were identified to be QS regulated. Expression studies revealed that Hfq does not control the master regulators of the Las system, LasR and LasI. Upon entry into stationary phase, Hfq exerted a moderate stimulatory effect on translation of the rhlR gene and on the qscR gene, encoding a LasR/RhlR homologue. However, Hfq considerably stimulated translation of the rhlI gene, encoding the synthetase of the autoinducer N-Butyryl-homoserine lactone (C4-HSL). Correspondingly, the C4-HSL levels were reduced in a hfq(-) strain. To elucidate the stimulatory effect of Hfq on rhlI expression we asked whether Hfq affects the stability of the regulatory RNAs RsmY and RsmZ, which have been implicated in sequestration of the translational repressor RsmA, which in turn is known to negatively regulate RhlI synthesis. We demonstrate that Hfq binds to and stabilizes the regulatory RNA RsmY, which is further shown to bind to the regulatory protein RsmA. A model for the Hfq regulatory network is presented, wherein an alleviation of the negative effect of RsmA accounts for the observed stimulation of rhlI expression by Hfq. The model is corroborated by the observation that a rsmY(-) mutant mimics the hfq(-) phenotype with regard to rhlI expression. PMID- 16468995 TI - Identification of a megaplasmid centromere reveals genetic structural diversity within the repABC family of basic replicons. AB - The basic replication unit of many plasmids and second chromosomes in the alpha proteobacteria consists of a repABC locus that encodes the trans- and cis-acting components required for both semiautonomous replication and replicon maintenance in a cell population. In terms of physical genetic organization and at the nucleotide sequence level, repABC loci are well conserved across various genera. As with all repABC-type replicons that have been genetically characterized, the 1.4 Mb pSymA and 1.7 Mb pSymB megaplasmids from the plant endosymbiont Sinorhizobium meliloti encode strong incompatibility (inc) determinants. We have identified a novel inc sequence upstream of the repA2 gene in pSymA that is not present on pSymB and not reported in other repABC plasmids that have been characterized. This region, in concert with the repA and repB genes, stabilizes a test plasmid indicating that it constitutes a partitioning (par) system for the megaplasmid. Purified RepB binds to this sequence and binding may be enhanced by RepA. We have isolated 19 point mutations that eliminate incompatibility, reduce RepB binding or the stabilization phenotype associated with this sequence and all of these map to a 16-nucleotide palindromic sequence centred 330 bp upstream of the repA2 gene. An additional five near-perfect repeats of this palindrome are located further upstream of the repA2 gene and we show that they share some conservation with known RepB binding sites in different locations on other repABC plasmids and to two sequences found on the tumour inducing plasmid of Agrobacterium tumefaciens. These additional palindromes also bind RepB but one of them does not display obvious incompatibility effects. A heterogenic distribution of par sequences demonstrates unexpected diversity in the structural genetic organization of repABC loci, despite their obvious levels of similarity. PMID- 16468996 TI - GTP drives myosin light chain 1 interaction with the class V myosin Myo2 IQ motifs via a Sec2 RabGEF-mediated pathway. AB - The yeast myosin light chain 1 (Mlc1p) belongs to a branch of the calmodulin superfamily and is essential for vesicle delivery at the mother-bud neck during cytokinesis due to is ability to bind to the IQ motifs of the class V myosin Myo2p. While calcium binding to calmodulin promotes binding/release from the MyoV IQ motifs, Mlc1p is unable to bind calcium and the mechanism of its interaction with target motifs has not been clarified. The presence of Mlc1p in a complex with the Rab/Ypt Sec4p and with Myo2p suggests a role for Mlc1p in regulating Myo2p cargo binding/release by responding to the activation of Rab/Ypt proteins. Here we show that GTP or GTPgammaS potently stimulate Mlc1p interaction with Myo2p IQ motifs. The C-terminus of the Rab/Ypt GEF Sec2p, but not Sec4p activation, is essential for this interaction. Interestingly, overexpression of constitutively activated Ypt32p, a Rab/Ypt protein that acts upstream of Sec4p, stimulates Mlc1p/Myo2p interaction similarly to GTP although a block of Ypt32 GTP binding does not completely abolish the GTP-mediated Mlc1p/Myo2p interaction. We propose that Mlc1p/Myo2p interaction is stimulated by a signal that requires Sec2p and activation of Ypt32p. PMID- 16468997 TI - Inactivation of the fibronectin-binding adhesin gene bbk32 significantly attenuates the infectivity potential of Borrelia burgdorferi. AB - Borrelia burgdorferi, the aetiological agent of Lyme disease, utilizes multiple adhesins to interact with both the arthropod vector and mammalian hosts it colonizes. One such adhesive molecule is a surface-exposed fibronectin-binding lipoprotein, designated BBK32. Previous characterization of BBK32-mediated fibronectin binding has been limited to biochemical analyses due to the difficulty in mutagenizing infectious isolates of B. burgdorferi. Here we report an alternative method to inactivate bbk32 via allelic exchange through use of a low-passage variant of B. burgdorferi strain B31 that is more readily transformed. The resulting mutant does not synthesize BBK32, exhibits reduced fibronectin binding in solid phase assays and manifests decreased interactions with mouse fibroblast cells relative to both the infectious parent and genetic complement. Furthermore, the bbk32 knockout was significantly attenuated in the murine model of Lyme disease, whereas a genetically complemented control was not, indicating that BBK32 is necessary for maximal B. burgdorferi infection in the mouse. To our knowledge this is the first mutational analysis of a surface exposed, functional borrelial lipoprotein adhesin whose activity is associated with the mammalian host environment. By analogy with other pathogens that utilize fibronectin binding as an important virulence determinant, the borrelial fibronectin-BBK32 interaction is likely to be important in B. burgdorferi specific pathogenic mechanisms, particularly in the context of dissemination, secondary colonization and/or persistence. PMID- 16468999 TI - PspB and PspC of Yersinia enterocolitica are dual function proteins: regulators and effectors of the phage-shock-protein response. AB - The phage-shock-protein (Psp) stress-response system is conserved in many bacteria and has been linked to important phenotypes in Escherichia coli, Salmonella enterica and also Yersinia enterocolitica, where it is essential for virulence. It is activated by specific extracytoplasmic stress events such as the mislocalization of secretin proteins. From studies of the Psp system in E. coli, the cytoplasmic membrane proteins PspB and PspC have only been proposed to act as positive regulators of psp gene expression. However, in this study we show that PspB and PspC of Y. enterocolitica are dual function proteins, acting both as regulators and effectors of the Psp system. Consistent with the current model, they positively control psp gene expression in response to diverse inducing cues. PspB and PspC must work together to achieve this regulatory function, and bacterial two-hybrid (BACTH) analysis demonstrated a specific interaction between them, which was confirmed by in vivo cross-linking. We also show that PspB and PspC play a second role in supporting growth when a secretin protein is overexpressed. This function is independent from their role as regulators of psp gene expression. Furthermore, whereas PspB and PspC must work together for their regulatory function, they can apparently act independently to support growth during secretin production. This study expands the current understanding of the roles played by PspB and PspC, and demonstrates that they cannot be considered only as positive regulators of psp gene expression in Y. enterocolitica. PMID- 16469000 TI - A C-terminal translocation signal is necessary, but not sufficient for type IV secretion of the Helicobacter pylori CagA protein. AB - Type IV secretion systems are increasingly recognized as important virulence determinants of Gram-negative bacterial pathogens. While the examination of several type IV-secreted proteins suggested that their secretion depends on C terminal signals, the nature of these signals and their conservation among different systems remain unclear. Here, we have characterized the secretion signal of the Helicobacter pylori CagA protein, which is translocated by the Cag type IV secretion apparatus into eucaryotic cells. The production of fusion proteins of CagA and green fluorescent protein (GFP) did not result in translocation of GFP to epithelial cells, but a fusion of GFP with the CagA C terminus exerted a dominant-negative effect upon wild-type CagA translocation. We show that CagA translocation depends on the presence of its 20 C-terminal amino acids, containing an array of positively charged residues. Interestingly, these positive charges are neither necessary nor sufficient for CagA translocation, but replacing the C-terminal region of CagA with that of other type IV-secreted proteins reconstitutes CagA translocation competence. Using a novel type IV translocation assay with a phosphorylatable peptide tag, we show that removal of the N-terminal part of the CagA protein renders the protein translocation incompetent as well. Thus, the Cag type IV secretion system seems to diverge from other systems not only with respect to its composition and architecture, but also in terms of substrate recognition and transport. PMID- 16468998 TI - A new protein superfamily includes two novel 3-methyladenine DNA glycosylases from Bacillus cereus, AlkC and AlkD. AB - Soil bacteria are heavily exposed to environmental methylating agents such as methylchloride and may have special requirements for repair of alkylation damage on DNA. We have used functional complementation of an Escherichia coli tag alkA mutant to screen for 3-methyladenine DNA glycosylase genes in genomic libraries of the soil bacterium Bacillus cereus. Three genes were recovered: alkC, alkD and alkE. The amino acid sequence of AlkE is homologous to the E. coli AlkA sequence. AlkC and AlkD represent novel proteins without sequence similarity to any protein of known function. However, iterative and indirect sequence similarity searches revealed that AlkC and AlkD are distant homologues of each other within a new protein superfamily that is ubiquitous in the prokaryotic kingdom. Homologues of AlkC and AlkD were also identified in the amoebas Entamoeba histolytica and Dictyostelium discoideum, but no other eukaryotic counterparts of the superfamily were found. The alkC and alkD genes were expressed in E. coli and the proteins were purified to homogeneity. Both proteins were found to be specific for removal of N-alkylated bases, and showed no activity on oxidized or deaminated base lesions in DNA. B. cereus AlkC and AlkD thus define novel families of alkylbase DNA glycosylases within a new protein superfamily. PMID- 16469001 TI - Apples and oranges. Re: 7.4-year update of the ongoing bicalutamide Early Prostate Cancer (EPC) trial programme. PMID- 16469002 TI - The third analysis of the bicalutamide Early Prostate Cancer programme. PMID- 16469003 TI - Dynamic variables: novel and perhaps better predictors of progression in benign prostatic hyperplasia. PMID- 16469004 TI - Y chromosome microdeletions and male infertility: who should be tested and why? PMID- 16469005 TI - Radical prostatectomy with positive surgical margins: how are patients managed? PMID- 16469006 TI - Current role of prostate-specific antigen kinetics in managing patients with prostate cancer. PMID- 16469007 TI - Management approaches to small renal tumours. PMID- 16469008 TI - Treatment targeted at vascular endothelial growth factor: a promising approach to managing metastatic kidney cancer. PMID- 16469009 TI - Functional outcomes and oncological efficacy of Vattikuti Institute prostatectomy with Veil of Aphrodite nerve-sparing: an analysis of 154 consecutive patients. AB - OBJECTIVES: To report updated results, at 1 year of follow-up, of a modified nerve-sparing robotic radical prostatectomy that preserves the lateral prostatic fascia (Veil of Aphrodite). PATIENTS AND METHODS: From January to December 2003, 154 consecutive men had a Vattikuti Institute prostatectomy with Veil of Aphrodite nerve-sparing by one surgeon. A prospective database recorded patient demographics, intraoperative, peri-operative, and pathological variables. Peri operative complications were recorded using the Clavien classification. Patients had serum prostate-specific antigen (PSA) levels measured every 3 months and self administered the International Prostate Symptom Score and Sexual Health Inventory for Male questionnaires before and at 1 year after surgery. RESULTS: The men had a mean age of 57.4 years, a mean body mass index of 27.2 kg/m(2), and a mean PSA level before surgery of 5.11 ng/mL. The mean operative duration was 122 min. At 1 year, 96% of the men reported having had intercourse and 71% had recovered normal erectile function. One man had a Clavien grade II complication, 4.6% of men with organ-confined disease had positive surgical margins, and no patient had a PSA recurrence at 12 months; 97% of the men were continent at 1 year, and the median time to continence was 14 days. CONCLUSION: Veil of Aphrodite nerve-sparing surgery provides better recovery of sexual function at 1 year than in contemporary series from centres of excellence, without compromising cancer control and urinary function. PMID- 16469010 TI - Positive influence of robotically assisted laparoscopic prostatectomy on the collaborative-care pathway for open radical prostatectomy. AB - OBJECTIVE: To define changes in the clinical pathway for open radical retropubic prostatectomy (RRP) and to determine their impact on patient care under the indirect influence of minimally invasive prostatectomy after implementing robotically assisted laparoscopic prostatectomy (RALP) at our institution and modifying the collaborative-care pathway. PATIENTS AND METHODS: We retrospectively reviewed the Vanderbilt Prostate Database since the initiation of RALP in May 2003, evaluating those patients who had open RRP. Data were analysed in three consecutive 5-month blocks to determine incremental changes in protocol and outcomes, including complications and re-admissions. RESULTS: Between May 2003 and August 2004, 196 patients had RALP and 183 had RRP. After May 2003, there was a progressive reduction for open RRP in the length of stay from 1.6 to 1.3 days. Analysis also showed an elimination of patient-controlled analgesia, a rapid diet advancement, and drain removal at the surgeon's discretion. There were no major complications after surgery, and three minor complications (1.6%). The rate of re-admission after open RRP was low throughout the study period (seven patients, 3.8%), and did not increase progressively. CONCLUSIONS: The management after open RRP was modified substantially after implementing RALP, with the two clinical pathways now similar. Despite these modifications in the treatment of patients having open RRP there was no increase in morbidity or compromise in patient outcome. Indeed, this study shows the feasibility and safety of a 1-day length of stay for open RRP. PMID- 16469011 TI - Recovery of serum testosterone after neoadjuvant androgen deprivation therapy and radical radiotherapy in localized prostate cancer. AB - OBJECTIVE: To prospectively evaluate the time-course of recovery of serum testosterone levels after a short course of luteinizing hormone-releasing hormone analogue (LHRHa) and radical radiotherapy to the prostate. PATIENTS AND METHODS: Testosterone, luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were sequentially measured prospectively in 59 men who received short-course LHRHa treatment and radiotherapy for localized prostate cancer. Measurements were made before treatment (baseline), during LHRHa treatment, and at 6, 12, 18, 24 and >40 weeks after the last LHRHa injection. RESULTS: The median (range) time from the first to last LHRHa injection was 116 (54-194) days. The mean (95% confidence interval) testosterone levels (in nmol/L) at baseline, during treatment and at 6, 12, 18, 24 and >40 weeks afterward were 12.0 (10.8-13.1), 0.6 (0.5-0.7), 1.4 (0.6-2.2), 11.4 (9.7-13), 12.2 (10.5-14), 10.4 (8.9-12) and 11.7 (10.5-13). Four men had low baseline testosterone levels (<6.1 nmol/L). At 6 weeks after the last LHRHa injection, no men had testosterone levels in the 'normal' range; 35% were in the normal range at 12 weeks, 85% at 18 weeks, 89% at 24 weeks, and 96% at 1 year. CONCLUSION: After LHRHa treatment and radiotherapy, the testosterone levels of most men had recovered to normal by 18-24 weeks after the last LHRHa injection. PMID- 16469012 TI - Is prostate-specific antigen (PSA) density better than the preoperative PSA level in predicting early biochemical recurrence of prostate cancer after radical prostatectomy? AB - OBJECTIVE: To investigate the relationship of preoperative prostate-specific antigen (PSA) level and PSA density with several clinical and pathological variables, including biochemical recurrence after radical prostatectomy (RP), and to compare the preoperative PSA level and PSA density as prognostic factors in prostate cancer. PATIENTS AND METHODS: The study included 348 patients who had a RP at one institution, with whole-mount specimens of the prostate examined by one pathologist. Univariate and multivariate analyses were used to assess the relationship of the preoperative PSA level and PSA density with clinical and pathological variables, and by receiver operating characteristic (ROC) analysis to evaluate the relative usefulness of the two factors as predictors for biochemical recurrence. RESULTS: The PSA level before RP was significantly correlated (Spearman's rank correlation) with patient age (P = 0.003), prostate weight (P < 0.001), cancer volume (P < 0.001) and Gleason score (P = 0.033), and with surgical margin status and pathological stage (both P < 0.001) in the RP specimen. In the multivariate analysis controlling for tumour stage, surgical margin status, and Gleason score, both PSA level and PSA density were significant predictors of PSA recurrence (P = 0.027 and 0.01, respectively). ROC analysis showed no statistical difference between the PSA level and PSA density in predicting PSA recurrence after RP (P = 0.40). CONCLUSIONS: These results show a significant correlation of the preoperative PSA level with other established prognostic factors for prostate cancer. In the multivariate analysis, both PSA level and PSA density were independent predictors of PSA recurrence. Because the PSA level is as effective as PSA density in predicting PSA recurrence, the extra effort required to calculate PSA density may not be warranted. We recommend that the PSA level before RP be considered in stratifying patients into different prognostic groups, and in determining the optimum management. PMID- 16469013 TI - Prostate carcinoma I: prognostic factors in radical prostatectomy specimens and pelvic lymph nodes. PMID- 16469014 TI - Prostate carcinoma II: prognostic factors in prostate needle biopsies. PMID- 16469015 TI - Changes in vascular flow after transdermal oestradiol therapy for prostate cancer: a mechanism for cardiovascular toxicity and benefit? AB - OBJECTIVE: To report the influence of transdermal oestradiol therapy on the vascular dynamics of men with advanced prostate cancer. PATIENTS AND METHODS: Twenty patients with newly diagnosed locally advanced or metastatic prostate cancer (10 each) were treated using transdermal oestradiol patches. The vascular flow was assessed 6-monthly before and during a year of therapy using arterial and venous Doppler and duplex ultrasonography, arterial and venous photoplethysmography and opto-electronic plethysmography. RESULTS: Arterial flow, as measured by the mean and peak systolic velocities and photoplethysmography, significantly increased over time. Arterial compliance initially decreased but had normalized after 12 months. The venous variables were unaffected. As a result, the total limb blood flow and the capillary filtration rate were significantly increased. CONCLUSION: Transdermal oestradiol therapy causes an increase in arterial but not venous flow, and an initial decrease in arterial compliance, which adapts to the physiological range with time. It is possible that these changes may account for the increase in cardiovascular toxicity seen in the early phase of oestrogen therapy, and the cardioprotective effect that accrues thereafter. PMID- 16469016 TI - Surgical treatment of adrenal metastasis from renal cell carcinoma: a single centre experience of 45 patients. AB - OBJECTIVE: To report, in a retrospective study, the diagnostic problems and oncological results of surgery in patients with either synchronous or metachronous adrenal metastasis, which are uncommon in renal cancer, at 2-10% of patients. PATIENTS AND METHODS: Of 1179 patients treated for renal cancer between 1987 and 2003, 914 had renal surgery with concomitant ipsilateral adrenalectomy (routinely in 875 and for abnormal findings on computed tomography, CT, in 39) and 15 contralateral adrenalectomy (all after suspicious findings on CT). During the follow-up after renal surgery, another 14 patients had adrenalectomy for CT evidence of an abnormal adrenal gland, contralateral to the previous renal tumour in 12 and bilaterally in two. RESULTS: Of 914 ipsilateral adrenal glands removed during renal surgery, 854 (93.5%) were normal on pathological examination, 28 (3%) had a benign pathology, six (0.8%) were directly infiltrated by the tumour and 26 (2.7%) were metastatic. For both benign and metastatic ipsilateral adrenal pathology, CT had sensitivity, specificity and positive/negative predictive values of 47%, 99%, 73% and 96%, respectively. Of 29 contralateral glands removed because of suspicious CT findings (15 at diagnosis of renal cancer, 14 during the follow-up) there was no abnormality in one (3.4%), a benign pathology in seven (24%) and a metastasis in 21 (72%). Thus there were 32 synchronous (incidence 2.7%; ipsilateral to the renal tumour in 24, contralateral in six and bilateral in two), and 13 metachronous adrenal metastases (incidence 1.0%; contralateral in 11 and bilateral in two). The metachronous metastases were diagnosed at a mean (range) interval of 30.6 (8-73) months after renal surgery. No ipsilateral adrenal metastases were discovered at diagnosis or during the follow-up in the 382 patients with an organ-confined renal tumour of <4 cm in diameter. Twenty seven patients with an isolated adrenal metastasis (synchronous in 14, metachronous in 13) had statistically significantly (P < 0.001) better survival than the 18 (all synchronous) with multiple sites of metastatic disease. In particular, there was long-term survival (mean 83 months) in 10 patients with an isolated adrenal metastasis. CONCLUSION: Sparing the ipsilateral adrenal is advisable only for organ-confined renal tumours of <4 cm in diameter; clinical local staging of renal cancer is the best predictor of the risk of adrenal metastasis. Conversely, CT had good diagnostic ability for the contralateral adrenal gland, especially during the follow-up. Some patients with isolated adrenal metastasis could be treated by metastasectomy, with long-term survival free of disease and confirming that, even if in a few and unselectable patients, removing all the neoplastic bulk can be curative. Nevertheless, the high rate of relapse underlines the need for an effective systemic therapy, and more so for widespread metastatic disease that currently cannot be cured. PMID- 16469017 TI - Immediate administration of intravesical mitomycin C after tumour resection for superficial bladder cancer. AB - OBJECTIVE: To assess the feasibility and safety of administering intravesical mitomycin C in theatre immediately after transurethral resection of bladder tumour (TURBT). PATIENTS AND METHODS: A protocol was developed to allow the safe administration of mitomycin C in theatre immediately after TURBT. Over a 32-month period all patients not excluded by the protocol were given mitomycin C in theatre after TURBT, and any adverse events reported. RESULTS: In all, 177 instillations were carried out; there were two minor patient-related complications, and no staff-related adverse events. CONCLUSION: The immediate administration of mitomycin C in theatre after TURBT is feasible and safe for patients and staff. It provides the earliest and surest prophylaxis against tumour cell re-implantation at TURBT. PMID- 16469018 TI - Long-term efficacy and safety of alfuzosin 10 mg once daily: a 2-year experience in 'real-life' practice. AB - OBJECTIVE: To assess the 2-year efficacy and safety of alfuzosin 10 mg once daily, a selective alpha(1)-adrenoceptor antagonist, in men complaining of lower urinary tract symptoms (LUTS) suggestive of benign prostate hyperplasia (BPH), in 'real life' practice. PATIENTS AND METHODS: In all, 839 European men with LUTS (mean age 67.3 years) were enrolled by general practitioners in a 2-year open label study with alfuzosin 10 mg once daily. They were asked to complete the International Prostate Symptom Score (IPSS), its appended eighth question (bother score), and the five domains (sexual drive, erection, ejaculation, problem assessment, and overall satisfaction) of the Brief Male Sexual Function Inventory (BSFI). The results were analysed at the endpoint in the intent-to-treat population. RESULTS: At the endpoint the total IPSS improved by 7 points (-38.5%) from baseline (P < 0.001) with 76.9% and 49.7% of men having an improvement of > or = 3 points and >6 points, respectively. There were also significant improvements in nocturia (-0.9, -30%; P < 0.001) and bother score (-1.8, -43%; P < 0.001) from baseline. Most patients (56%) perceived symptom relief within the first 2 weeks of treatment. All BSFI domains significantly improved from baseline (P < 0.05; <0.001 for overall satisfaction) and these improvements were more marked in men with severe LUTS at baseline. Alfuzosin 10 mg was well tolerated; the most common adverse event related to vasodilatation was dizziness/postural dizziness (3.1%). Ejaculatory disorders were uncommon (0.3%). Changes in blood pressure remained marginal, including in elderly men and those receiving antihypertensive agents. CONCLUSIONS: Alfuzosin 10 mg administered for 2 years in real practice is effective in improving LUTS and quality of life, and is well tolerated from a cardiovascular perspective, including in elderly men and those receiving antihypertensive co-medication. Ejaculatory disorders are uncommon. Alfuzosin may even slightly improve various domains of sexual function, such as sexual drive, erection, ejaculation and satisfaction with sexual life. PMID- 16469019 TI - A double-blind randomized dose-response study comparing daily doses of 5, 10 and 15 mg controlled-release oxybutynin: balancing efficacy with severity of dry mouth. AB - OBJECTIVE: To assess the efficacy, incidence of dry mouth and overall satisfaction with initial doses of 5, 10 and 15 mg of a new, once-daily, controlled-release (CR) form of oxybutynin for treating urge urinary incontinence (UUI). PATIENTS AND METHODS: Patients who reported urinary incontinence (UI) (one or more episodes/diary) and voiding frequency (eight or more voids/day) or urgency (one or more episodes/diary) during a 2-week baseline were randomized to once-daily 5, 10 or 15 mg CR oxybutynin for 4 weeks. Daily episodes of UI, voids, urgency, adverse events, dry mouth and satisfaction were recorded in a 3-day diary at baseline and after 4 weeks of treatment. In all, 237 patients were randomized and evaluated. RESULTS: Episodes of UI, voids and urgency were significantly reduced over the study period at all doses. Daily UI episodes were significantly lower with 15 mg/day than 5 and 10 mg/day. Dry mouth symptoms were similar in the 10 and 15 mg/day groups, and higher than in the 5 mg/day group. However, significantly greater overall satisfaction was reported with 15 than 5 mg/day. CONCLUSIONS: There were significant dose-response relationships with CR oxybutynin for both UI episodes and dry mouth. The greatest satisfaction was with 15 mg/day, and the severity of dry mouth was comparable at 10 mg/day, indicating that greater efficacy at the higher dose did not compromise tolerability. PMID- 16469020 TI - Predictive factors for nocturia in elderly men: a cross-sectional study in 21 general practices. AB - OBJECTIVE: To measure the prevalence of nocturia in general practice and to determine which factors are associated with nocturia. SUBJECTS AND METHODS: Data were collected from 3048 elderly men, who completed a questionnaire that was sent to every man aged 55-75 years in 21 general practices in Maastricht (the Netherlands). The symptom of nocturia was defined as two or more nocturnal voids. We investigated the prevalence of nocturia and the predictive relationships with the following factors: cerebrovascular disease, diabetes mellitus/insipidus (DMI), Parkinson's diseases, cardiovascular disease, hypertension, bladder/prostate cancer, kidney diseases, urinary bladder inflammation, congenital diseases (kidneys or prostate), using medical treatment for lower urinary tract symptoms, other treatment, psychological depression, symptoms suggestive of benign prostatic hyperplasia (BPH), and alcohol intake. RESULTS: Data from 2934 respondents were analysed; the prevalence of nocturia (two or more nocturnal voids) was 32.9% (965 men). The frequency of the number of nocturnal voids was: zero in 588 (20.0%), one in 1344 (45.8%), two in 611 (20.8%), three in 208 (7.1%), four in 70 (2.4%), and five or more in 76 (2.6%), with 37 values missing. A multivariate logistic regression analysis showed that nocturia in elderly men was significantly related to bladder/prostate cancer, cerebrovascular disease, treatment of voiding disorders, and moderate alcohol consumption. Next to these, BPH had a significant relationship with nocturia, especially in respondents with DMI and hypertension. Cardiovascular disease or hypertension was significantly related to nocturia, mutually replacing each other as a risk factor. CONCLUSION: Nocturia in elderly men is be related to many sources of potential risk factors: earlier urological diseases, cardiovascular and cerebrovascular diseases, BPH, DMI and behavioural habits. Some of these sources may interact and generate especially high risk in some groups for nocturia. PMID- 16469021 TI - An adjustable male sling for treating urinary incontinence after prostatectomy: a phase III multicentre trial. AB - OBJECTIVE: To evaluate the efficacy and safety of a new adjustable bulbourethral sling (Argus, Promedon SA, Cordoba, Argentina) in the treatment of male stress urinary incontinence (SUI) after prostate surgery. PATIENTS AND METHODS: In all, 48 patients with SUI because of prostatic surgery for prostate cancer (39) or benign prostatic hyperplasia (nine) had a new sling implanted in a multicentre trial at six institutions between April 2003 and September 2004. All patients were fully evaluated, including a questionnaire (International Consultation on Incontinence Questionnaire-Short Form, ICIQ-SF, range 0-21), endoscopy, and urodynamic evaluation. The Argus system comprises a 4.2 x 2.6 x 0.9 cm thick silicone foam pad for soft bulbar urethral compression. The pad is attached to the silicone cone columns that, after being passed with needles from the perineum to the abdominal wall, are adjusted with silicone washers to regulate and keep the desired tension against the urethra. The pad and washers are radio-opaque, which allows their position to be assessed during follow-up. The surgical technique was one described previously, with some modifications. RESULTS: At a mean (range) follow-up of 7.5 (1-17.5) months, 35 (73%) of the 48 patients were dry, five (10%) were improved, and eight (17%) were incontinent, including four (8%) who needed sling adjustment. The mean (range) ICIQ-SF improved from 19.2 (12 21) to 4 (0-21). There were three (6%) urethral perforations during surgery that were resolved by re-passing the needle. The sling was removed in three men (6%) due to erosion and in two (4%) due to infection. Seven (15%) cases of acute urinary retention resolved spontaneously, except for one that needed the sling loosening. No cases of chronic retention were reported. There was perineal discomfort and mild dysuria soon after surgery that resolved spontaneously after a few weeks. CONCLUSION: This new adjustable male sling safely and effectively controls sphincter incontinence in men after prostate surgery, with an acceptably low complication rate. The early results are encouraging; the Argus is a valid alternative to the artificial urinary sphincter, the standard therapy for this condition. PMID- 16469022 TI - Time to onset of improvement in symptoms of overactive bladder using antimuscarinic treatment. AB - OBJECTIVE: To evaluate the time to onset of statistically significant and clinically meaningful effects of trospium chloride in patients with an overactive bladder (OAB). PATIENTS AND METHODS: Data from a recent Phase III clinical study of trospium chloride were obtained, in which 658 patients with OAB were randomized (1 : 1) to placebo or trospium chloride 20 mg twice daily for 12 weeks. Original study safety endpoints consisting of adverse events, vital signs, electrocardiograms, and laboratory changes were collected, while original efficacy endpoints included number of toilet voids/day, urgency severity per toilet void, urge urinary incontinence (UUI), and volume voided per toilet void at weeks 1, 4, and 12. Results were also analysed using the OAB Symptom Composite Score (OAB-SCS). The efficacy analysis focused on changes from baseline in these endpoints from 1 to 7 days of treatment to establish the earliest point at which there was statistical significance or clinical efficacy. RESULTS: There were statistically significant improvements in efficacy over placebo in endpoints (toilet voids, urgency severity/void, UUI episodes, OAB-SCS) within a few days of treatment, with improvements in symptoms continuing to 7 days of treatment. There were clinically meaningful improvements in most endpoints by the end of the first week. CONCLUSIONS: The time to onset of the clinical effect should be studied more extensively to identify when patients might expect a clinically meaningful improvement in their OAB-related symptoms. PMID- 16469023 TI - Percutaneous tibial nerve stimulation in the treatment of refractory overactive bladder syndrome: is maintenance treatment necessary? AB - OBJECTIVE: To determine the effect of a pause in percutaneous tibial nerve stimulation (PTNS) in successfully treated patients with an overactive bladder (OAB), and the reproducibility of successful treatment when restored. PATIENTS AND METHODS: Eleven patients (mean age 51 years) with refractory OAB (more than seven voids and/or three or more urge incontinence episodes per day) were successfully treated with PTNS, and then discontinued treatment. Patients completed bladder diaries and quality-of-life (QoL) questionnaires (Short Form-36 and I-QoL) before (T1) and after a 6-week pause (T2) of maintenance PTNS, and again after re-treatment (T3). The first objective was defined as a > or = 50% increase in the incontinence episodes and/or voiding frequency in the bladder diary after T2. The second objective was defined as > or = 50% fewer incontinence episodes and/or voiding frequency in bladder diary after T3. RESULTS: At T2, seven of the 11 patients had a > or = 50% increase in incontinence episodes and/or voiding frequency in the bladder diary. The mean voided volume, nocturia, number of incontinence episodes and incontinence severity deteriorated significantly (P < 0.05). At T3, nine patients had > or = 50% fewer incontinence episodes and/or voiding frequency in the bladder diary. Nocturia, the number of incontinence episodes, incontinence severity, mean voided volume and quality of life improved significantly (P < 0.05). CONCLUSIONS: Continuous therapy is necessary in patients with OAB treated successfully by PTNS. The efficacy of PTNS can be reproduced in patients formerly treated successfully. PMID- 16469024 TI - Medical visits among adults with symptoms commonly associated with an overactive bladder. AB - OBJECTIVES: To examine nationally representative data and thus obtain estimates of the use of healthcare providers associated with the overactive bladder (OAB) symptoms, a condition characterized by frequency, urgency and nocturia, with or with no urge incontinence, as although it is ranked among the 10 most common chronic medical conditions in the USA, the level of OAB-associated medical treatment remains largely unknown. METHODS: To estimate the number of annual OAB associated medical visits among patients aged > or =18 years, three national databases in the USA (year 2000) were examined: the National Ambulatory Medical Care Survey, the National Hospital Ambulatory Medical Care Survey, and the National Hospital Discharge Survey. Population estimates were constructed using design-based statistical analyses to account for the complex survey designs of data. RESULTS: During 2000, adult Americans made 1.4 million (95% confidence interval 1.1-1.8 million) ambulatory visits to non-Federal office-based physicians with International Classification of Disease (ICD-9) coding indicative of OAB symptoms. Accounting for emergency and outpatient department visits, as well as non-Federal short-stay hospital discharges, the estimated number of medical visits with OAB-associated ICD-9 coding was <1.5 million. CONCLUSION: The prevalence of OAB was estimated to be 34 million adult Americans. When 1.4 million ambulatory visits were compared with this prevalence, as few as 4% of adult Americans with OAB sought medical treatment during the year 2000. The present results therefore suggest a large unmet medical need among the population of adult Americans with OAB. PMID- 16469025 TI - Frequency and nocturia after successful renal transplantation: a normal situation? AB - OBJECTIVE: To analyse lower urinary tract function before and after successful renal transplantation and compare the data with those from a healthy control group. PATIENTS AND METHODS: Data were gathered by retrospective analysis of 331 charts of patients transplanted between March 1998 and May 2003, using written questionnaires and personal interview, and investigation of 150 patients. The control group consisted of 150 urologically healthy volunteers. RESULTS: Frequency and nocturia were the main lower urinary tract symptoms. Frequency of more than six voids/day was reported by 87% and nocturia of more then one void/night by 93% of all patients after successful renal transplantation. There was no significant correlation with fluid intake, diuretic medication, gender or age. Over the years the number of voids tended to decrease but remained higher than in the control group. However, 94% of all patients were happy with the quality of life after renal transplantation. CONCLUSION: Frequency and nocturia are the two main characteristics of lower urinary tract function after renal transplantation, probably through a combination of high fluid intake, a long-term defunctionalized urinary bladder during renal replacement therapy, a denervated donor kidney, concomitant diseases and psychosocial distress. Quality of and satisfaction with life were not compromised. PMID- 16469026 TI - Beneficial effects of extended-release doxazosin and doxazosin standard on sexual health. AB - The prevalence of benign prostatic hypertrophy (BPH) and erectile dysfunction (ED) both increase with age, and increasing evidence suggests a common cause rather than independent age-related changes. Arterial hypertension often accompanies these urological disorders, suggesting the possibility that increased alpha-adrenoceptor activity may be causal in all three conditions. As evidence for this model, alpha-adrenoceptor antagonists such as doxazosin produce therapeutically beneficial effects in lowering blood pressure, reducing prostate growth and BPH symptoms, and relieving ED. At postjunctional alpha(1)-receptors in the corpus cavernosa, noradrenaline causes vascular smooth muscle cell contraction, restricting blood flow, resulting in penile detumescence. Just as alpha-adrenoceptor antagonism results in systemic vasorelaxation to lower blood pressure, the same mechanism in the penis modulates the effects of noradrenaline to favour vasodilatation, resulting in improved erectile function. Increasing clinical evidence attests to the effectiveness of doxazosin in relieving ED, even in patients refractory to ED-specific treatment, as well as in reducing BPH symptoms and elevated blood pressure. PMID- 16469027 TI - Comparative analysis of the penis corpora cavernosa in controls and patients with erectile dysfunction. AB - OBJECTIVE: To evaluate the presence of structural disorders of the corpora cavernosa in patients with erectile dysfunction (ED), as despite new drugs being effective in many men with ED, some aspects of structural disorders of the corpora cavernosa remain unknown. MATERIALS AND METHODS: Biopsy specimens were taken from the corpora cavernosa of seven patients (mean age 57.8 years, range 51 72) with severe ED who had a penile prosthesis implanted. The controls tissues were fragments of corpora cavernosa obtained from autopsies of six men (mean age 52.3 years, range 40-66) who died from causes unrelated to the urogenital system. For light microscopy, the specimens were processed routinely to paraffin wax, and by immunohistochemistry to evaluate elastic fibres, and by Masson's trichrome to analyse collagen and smooth muscle fibres. Stereological methods were used to quantitatively evaluate the different elements (as a percentage). RESULTS: The percentages of the different elements in the human penis of controls and men with ED, respectively, were: elastic fibres 13.2% and 9.1%; collagen fibres 40.8% and 41.6%; and smooth muscle, 40.4% and 42%. CONCLUSIONS: In patients with ED there was a statistically significant reduction in the percentage of elastic fibres, but no statistically significant difference in collagen and smooth muscle fibres, and no appreciable differences in collagen distribution between the groups. PMID- 16469028 TI - Risk factors for Peyronie's disease: a case-control study. AB - OBJECTIVE: To test some hypotheses about risk factors for Peyronie's disease (PD). PATIENTS AND METHODS: In a case-control study, 82 patients with PD, consecutively diagnosed at the first author's institution, were compared with 246 men visiting the same institution for dermatological diseases. Univariate and multivariate logistic regression analyses were used to assess the data. RESULTS: From the multivariate logistic regression analysis the risk factors for PD were: a history of genital and/or perineal injuries, transurethral prostatectomy, cystoscopy, diabetes mellitus, hypertension, lipoma, propranolol in therapy, Dupuytren's contracture in the medical history, ever having smoked, alcohol consumption, fibromatous lesions of the genital tract of the partner, and surgical intervention on the genital tract of the partner. CONCLUSION: The results of the present study are in line with the hypothesis that, in addition to genetic predisposition, trauma of the penis and systemic vascular diseases are risk factors for PD. Smoking and alcohol consumption also seem to have some role in the development of the disease. PMID- 16469029 TI - Early patency rates after the two-suture invagination technique of vaso epididymal anastomosis for idiopathic obstruction. AB - OBJECTIVES: To prospectively evaluate the results of vaso-epididymostomy, using a two-suture microsurgical invagination technique, when used for men with azoospermia due to an obstruction at the vaso-epididymal junction. PATIENTS AND METHODS: Between December 2002 and November 2004, 29 men with idiopathic obstructive azoospermia underwent vaso-epididymostomy with the two-suture invagination technique. The patency rate was assessed by return of sperm in the semen after surgery. RESULTS: In all, 23 men provided at least one postoperative semen sample. The mean (range) follow-up was 7.6 (1.5-30) months. In 11 of these men (48%), patency was shown at a mean of 3.2 (1.5-7) months after surgery. The median sperm density was 17 (10-65) million/mL. Four men had normal sperm density and motility (>20 million/mL; >50% total motility). CONCLUSIONS: Men with idiopathic vaso-epididymal junction obstruction can have significant sperm positivity after vaso-epididymostomy. With a patency rate of nearly half within a year of surgery, vaso-epididymostomy should be the first line of therapy for these men. PMID- 16469030 TI - Laparoscopic robotic-assisted management of pelvi-ureteric junction obstruction in patients with horseshoe kidneys: technique and 1-year follow-up. AB - OBJECTIVE: To report our experience with laparoscopic robotic-assisted management of pelvi-ureteric junction obstruction (PUJO) in patients with horseshoe kidneys. PATIENTS AND METHODS: Between January 2002 and May 2003, two men and a woman with horseshoe kidneys (mean age 44.6 years) had laparoscopic dismembered pyeloplasty with robotic assistance for PUJO with no division of the isthmus. Two patients had renal stones which were extracted during surgery. None of the patients had had previous abdominal surgery. RESULTS: The mean operative duration was 148.3 min, the mean estimated blood loss was <100 mL and the mean hospital stay was 7.6 days. Renal function was preserved in all three patients during the immediate and long-term follow-up as measured by intravenous urography. The three patients had durable clinical and radiographic success during a mean follow-up of 21 months. One patient needed complementary extracorporeal shockwave lithotripsy, and one had an episode of pyelonephritis, which was treated successfully. There were no other significant complications before or after surgery. CONCLUSION: Laparoscopic robotic-assisted pyeloplasty for horseshoe kidney is safe and feasible, offering the advantages of minimally invasive surgical procedures with enhanced laparoscopic skills related to the use of the robot. PMID- 16469031 TI - Magnetic resonance renal angiography and venography: an analysis of 111 consecutive scans before donor nephrectomy. AB - OBJECTIVE: To determine the accuracy of magnetic resonance imaging (MRI) renal angiography in predicting vascular anatomy before donor nephrectomy, to determine the significance of missed vessels and to ascertain whether vessels are missed because of technical limitations or errors in interpretation. PATIENTS AND METHODS: In all, 111 consecutive living donations were assessed; the anatomy on MRI before donation was compared with that at nephrectomy. The significance of additional arteries and veins was recorded at the time of donation, with extra vessels either anastomosed or sacrificed. Finally, the scans in which extra vessels had not been identified were re-examined to establish whether these could be identified retrospectively. RESULTS: In all, 93 kidneys had a single renal artery and 18 had two. All lower pole arteries were anastomosed and all upper pole arteries were sacrificed. Nine arteries were identified before surgery (five were to the lower pole), and nine were missed (four to the lower pole). There were 13 kidneys with more than one vein. Four of these were seen on MRI. However, an extra vein was anastomosed in only one case. On review of the imaging, three arteries were missed because of human error and six due to technical limitations. Of the nine missed veins, only three were easily identified retrospectively. Overall, using MRI as a preoperative investigation for the 111 consecutive cases, the surgeon encountered a previously unidentified accessory artery in nine (8%), and this required anastomosis in four (4%). CONCLUSION: MR angiography has the advantage over computed tomography (CT) of having virtually no side-effects, and if the small possibility is accepted of missing extra vessels because of technical limitation or interpretation, it is a good investigation. However, in light of the failure to visualize all arteries transplanted, we have started to use multi-slice (16-channel) CT to see if its improved spatial resolution alters the results. PMID- 16469032 TI - A modified sigma-rectum pouch (Mainz pouch II) technique: analysis of outcomes and complications on 220 patients. AB - OBJECTIVE: To evaluate the safety and efficacy of a modified technique for a sigma-rectum pouch (Mainz pouch II). PATIENTS AND METHODS: Between October 1994 and March 2003, a modified Mainz pouch II was constructed in 220 patients (153 men and 67 women, mean age 56.7 years) in a multicentre prospective study. The median follow-up (177 patients, 81%) was 21 (1-84) months. RESULTS: There were no peri-operative deaths; there were early complications in 24 patients, including prolonged ileus in six, pyelonephritis in 17, unilateral ureterohydronephrosis in 11, bilateral ureterohydronephrosis in two, and incipient renal failure in five. Late complications included ureteric implantation-site stenosis (unilateral in eight patients and bilateral in three). In all, 92 patients (52%) needed oral alkalizing medications and potassium supplementation because of hyperchloraemic metabolic acidosis. All but three patients (99%) were continent by day and at night. The mean (sd) voiding frequency was 4.2 (1.6) voids by day and 2.1 (0.5) at night. CONCLUSION: The Mainz pouch II is a simple and reproducible surgical technique, with good results in terms of mortality, morbidity, continence and overall quality of life. The main limitation is malignant transformation of the uretero-intestinal anastomosis. For selected cases, the technique is a good alternative to other types of continent urinary diversion. PMID- 16469033 TI - The 'learning curve' in hypospadias surgery. AB - OBJECTIVE: To provide an insight into the 'learning curve' of fellowship-trained paediatric urologists associated with hypospadias repair, as hypospadias surgery is one of the most common yet difficult procedures used by the paediatric urologist. PATIENTS AND METHODS: Prospective data were collected on 231 consecutive hypospadias operations performed by one paediatric urologist (M.H.) over a 5-year period, beginning with his first year after completing his fellowship. All patients were having their first surgery and none had a staged repair. Fistula formation was used as a surrogate for the complication rate, as it is an objective measurable outcome that is easily identified with little interobserver or parental/physician variability. The follow-up included several visits in the 15 months after repair, during which virtually all complications could be identified and addressed. RESULTS: The operative results improved throughout the 5 years of observation; there was a statistically significant decline in the fistula rate in each year of observation (P < 0.001; Kruskal Wallis exact test for ranked groups). The absolute reduction in fistula rates between the first 2 and the last 2 years was 12.7% (P < 0.02; chi squared). CONCLUSIONS: The science and surgery of hypospadiology is mostly and correctly delegated to the paediatric urologist. Even in the hands of a fellowship-trained paediatric urologist, a successful repair, as measured by complication rate, statistically improves with time and experience. PMID- 16469034 TI - Y-type urethral duplication in the male. AB - OBJECTIVE: To present a single series of Y-type duplication with an analysis of the presenting features, the management and outcome, as complete Y-type urethral duplication in the male is a rare congenital anomaly that presents many challenges. PATIENTS AND METHODS: We retrospectively reviewed patient charts, identifying those with urethral duplication, and then those with complete Y-type duplication. The age at presentation, investigations, management and outcomes were reviewed. RESULTS: Of 49 patients with a diagnosis of urethral duplication, 13 had the complete Y-type. Three were lost to follow-up and were therefore excluded from the study. The median (range) age at presentation was 2 months (birth to 10 years). All patients had other comorbidities, with anorectal malformations in seven, renal agenesis in four, sacral agenesis/partial agenesis in two, and unilateral/bilateral cryptorchidism in four. Presenting features included passage of urine perineally or rectally in seven, and dribbling from the orthotopic urethral meatus in one. The diagnosis was confirmed by micturating cysto-urethrography. A mean of 14 procedures was required to achieve a final complete repair, and included urethral reconstruction, repair of fistulae, urethral dilatation, and evaluative cystoscopy. Excluding cystoscopy and urethral dilatation, a mean (range) of 3 (1-5) reconstructive procedures was required in each patient. The median follow-up was 8 years. Five boys developed urethral strictures and one developed multiple fistulae. The best outcomes were in boys who had a staged urethral reconstruction. CONCLUSION: Due to the strong association with other congenital anomalies we recommend that all patients should be evaluated thoroughly, including a detailed physical examination, renal tract ultrasonography and spinal radiography. Surgical management remains a significant challenge, requiring many procedures with unpredictable outcomes, the goals of which should be to maintain continence and reconstruct the urethra with good cosmesis. The optimum management scheme must be individualized in this rare condition. PMID- 16469035 TI - A new fast-melting oral formulation of desmopressin: a pharmacodynamic study in children with primary nocturnal enuresis. AB - OBJECTIVE: To determine the pharmacodynamic properties of a new oral lyophilisate formulation of desmopressin (in single doses of 30, 60, 120, 240, 360 or 480 microg) in children with known primary nocturnal enuresis (PNE) and thus identify those dosages that could provide a duration of action corresponding to a typical length of night-time sleep in children with PNE; additional objectives were to determine the safety and tolerability of desmopressin in this population. PATIENTS AND METHODS: Children with PNE (mean three or more wet nights/week), aged 6-12 years, were randomized into a double-blind, placebo-controlled study. An overhydration technique was used before dosing to suppress endogenous vasopressin production and thereby ensure that any antidiuresis could be attributed to treatment. Dosing with desmopressin or placebo occurred when urinary production was >0.13 mL/min/kg. Urinary volume, osmolality and duration of urinary-concentrating action (above three threshold levels: 125, 200 and 400 mOsm/kg) were determined as endpoints. RESULTS: All 72 participants receiving desmopressin had a pharmacodynamic response to the drug, while there was no change in urinary output in the 12 placebo-treated patients. There was a clear relationship between desmopressin dose and duration of action and osmolality during action, although the three highest-dose groups had similar results. The mean duration of action of desmopressin at the lowest osmolality threshold level was 3.6-10.6 h, according to dose; for the highest threshold, the values were 1.3 8.6 h. CONCLUSION: Desmopressin, as the oral lyophilisate, causes a marked decrease in urinary output in hydrated children with PNE. A small dose range (120 240 microg) is likely to control diuresis for a period corresponding to a night's sleep (7-11 h) in most children with PNE. However, some patients might require a higher dose to obtain antidiuresis for the complete night. PMID- 16469036 TI - The functional effects of a c-kit tyrosine inhibitor on guinea-pig and human detrusor. AB - OBJECTIVE: To describe the effect of a specific c-kit receptor inhibitor (imatinib mesylate) on human detrusor strips in vitro and guinea-pig cystometry in vivo, and to show histological data suggesting differences in the distribution of interstitial cells of Cajal (ICC)-like cells in 'normal' and overactive human detrusor, as these cells have been identified as possible mediators of spontaneous activity and excitability in bladder smooth muscle. MATERIALS AND METHODS: Specimens of human detrusor were stained immunohistochemically with a c kit antibody. Human detrusor strips were mounted in a superfused organ-bath apparatus, and smooth muscle contraction was evoked with carbachol and electrical field stimulation in the presence and absence of imatinib mesylate. Also, guinea pig urodynamic studies were conducted before and after i.v. administration of imatinib mesylate, and changes in bladder variables and spontaneous activity were recorded. RESULTS: Imatinib mesylate (10(-6)M) inhibited evoked smooth muscle contraction and spontaneous activity in overactive human detrusor, with less effect on normal human tissue. Imatinib mesylate (10(-5)M) improved bladder capacity, compliance, voided volumes, urinary frequency, and reduced contraction thresholds and spontaneous activity during guinea-pig cystometry. c-kit labelling showed significantly more ICC-like cells in overactive human detrusor than in normal specimens. CONCLUSION: c-kit receptor blockers have inhibitory effects on guinea-pig and overactive human detrusor, possibly via c-kit receptors on bladder ICC-like cells. This and the possibility that there are more ICC-like cells in overactive bladder suggest that the c-kit receptor may provide a novel target for treating detrusor overactivity. PMID- 16469037 TI - Oral treatment with a vitamin D3 analogue (BXL628) has anti-inflammatory effects in rodent model of interstitial cystitis. AB - OBJECTIVE: To investigate the effects of a vitamin D3 analogue (BXL628) in a model of chronic cystitis, as calcitriol analogues might be an interesting new therapeutic option for interstitial cystitis, for although the cause of the disease remains unclear, the increase in mast cells in the mucosa and detrusor muscle are significant. MATERIALS AND METHODS: We devised a mouse model of allergen-induced allergic cystitis that is associated with the up-regulation of genes for interleukin-13, FcepsilonRIalpha and mast cells-derived proteases, a massive inflammatory reaction in the bladder tissue, and augmented levels of mast cell-derived protease 1 (MMCP1) detected in mouse sera. RESULTS: Oral administration of BXL628 significantly reduced the expression of interleukin-13, FcepsilonRIalpha and MMCP1 in the bladder. Furthermore, histological analysis showed a decrease in oedema and leukocyte infiltration in the bladder wall. BXL628 treatment reduced serum MMCP1 levels, indicating an effect on mast cell degranulation in vivo. CONCLUSIONS: Vitamin D3 analogues may successfully be used as anti-inflammatory agents in allergen-mediated inflammatory reactions. Moreover, the modulatory effect shown on mast cell activation by the BXL628 analogue strongly supports its potential therapeutic use in a possibly mast cell dependent disease such as human interstitial cystitis. PMID- 16469038 TI - Effects of long-term vardenafil treatment on the development of fibrotic plaques in a rat model of Peyronie's disease. AB - OBJECTIVES: To determine whether the phosphodiesterase-5 (PDE5) inhibitor, vardenafil, given orally and in different regimens, has a similar effect to that of the PDE5 inhibitor sildenafil, which prevented the development of a Peyronie's disease (PD)-like plaque formation induced by injecting transforming growth factor beta1 (TGF-beta1) into the tunica albuginea of the rat. MATERIALS AND METHODS: Vardenafil was given to male rats (eight per group) either in the drinking water or as an oral instillation once daily, at approximately 1 and approximately 3 mg/kg/day for 45 days after one injection with TGF-beta1 into the tunica albuginea, as an 'early preventive' treatment for TGF-beta1-induced formation of a PD-like plaque. Other groups received the two doses of vardenafil only in the drinking water, starting with a well-formed plaque, for 42 days ('late, therapeutic' administration). Sections of penile tissue were stained histochemically or immunohistochemically, followed by quantitative image analysis for collagen/smooth muscle and collagen III/I ratios, myofibroblast content (alpha-smooth muscle actin), TGF-beta1 expression, and apoptotic index. RESULTS: Preventative treatment with vardenafil at the higher dose (both continuous and once-daily treatments) reduced the collagen/smooth muscle and collagen III/I ratios, and the numbers of myofibroblasts and TGF-beta1-positive cells, and selectively increased the apoptotic index in the PD-like plaque. The lower dose was less effective, When vardenafil was given continuously in the drinking water for 41 days after the PD-like plaque was formed, there was only a partial reduction of the plaque. CONCLUSIONS: Long-term oral treatment with vardenafil slows and reverses the early stages of an experimental PD-like plaque in the rat, and might ameliorate a more advanced plaque. PMID- 16469039 TI - The effect of serotonin and serotonin antagonists on bladder cancer cell proliferation. AB - OBJECTIVE: To investigate the role of serotonin (5-hydroxytryptamine, 5HT) and its antagonists in the proliferation of high-grade bladder cancer cells (HT1376), as high-grade bladder cancer has a rapid rate of progression, invasion and recurrence, and 5HT antagonists inhibit the growth of the prostate cancer cell line (PC3). MATERIALS AND METHODS: HT1376 (human grade III transitional cell carcinoma) cells were incubated with either 5HT or 5HT antagonists (5HT(1A), 5HT(1B), 5HT(1D), 5HT(2), 5HT(3) and 5HT(4)). After 72 h, cell viability was assessed using the crystal violet assay. The presence of 5HT receptor subtypes on HT1376 cells and sections of human bladder cancer tissue was determined by immunohistochemistry and Western blot analysis. RESULTS: 5HT caused a dose dependent increase in the proliferation of HT1376 cells. The maximum increase in cell proliferation (12%; 12 samples, P < 0.001) was at 10(-8)m as compared to the control at 72 h. At 10(-4)m, 5HT(1A) antagonist (NAN-190 hydrobromide) and 5HT(1B) antagonist (SB224289 hydrochloride) had a 10% (12 samples, P < 0.001) and 93% (12, P < 0.001) inhibitory effect on HT1376 cell growth, respectively, compared to the control at 72 h. There was immunostaining for 5HT(1A) and 5HT(1B) receptors in HT1376 cells and malignant bladder tissue, confirming the presence of these two receptor subtypes. Western blot analysis showed the presence of 5HT(1A) and 5HT(1B) receptor proteins with bands of 46 kDa and 43 kDa, respectively. CONCLUSION: 5HT(1A) and to a greater extent 5HT(1B) antagonists significantly inhibit bladder cancer cell growth. This effect is probably mediated via the 5HT(1A) and 5HT(1B) receptors. These results highlight the potential use of 5HT(1A) and 5HT(1B) antagonists in the treatment of bladder cancer. PMID- 16469040 TI - Chemopreventive effects of cyclooxygenase-2 inhibitor and epidermal growth factor receptor kinase inhibitor on rat urinary bladder carcinogenesis. AB - OBJECTIVE: To examine the chemopreventive effects of a selective cyclooxygenase (COX)-2 inhibitor, meloxicam, and a selective epidermal growth factor (EGF) receptor tyrosine kinase inhibitor, gefitinib (as a single agent) on a carcinogen induced rodent bladder carcinogenesis model. MATERIALS AND METHODS: The study comprised 103 male Fisher-344 rats (8 weeks old); after initial carcinogen treatment for 8 weeks with 0.05%N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) in drinking water, the rats were divided into five groups, i.e. group 1, control (vehicle only); group 2, gefitinib high-dose (15 mg/kg by gavage once daily); group 3, gefitinib low-dose (5 mg/kg); group 4, meloxicam high-dose (1.8 mg/kg by gavage once daily); and group 5, meloxicam low-dose (0.6 mg/kg). Twelve weeks later the rats were killed; after fixing the bladder in 10% formalin, the number and size of hyperplasia and carcinoma foci were recorded microscopically in sections stained with haematoxylin and eosin, submitted entirely as multiple strips. RESULTS: The incidence of carcinoma, confirmed microscopically, was: control 14/20 (70%); high-dose gefitinib, 7/20 (35%); low-dose gefitinib, 7/20 (35%); high-dose meloxicam 7/21 (33%); and low-dose meloxicam, 12/20 (60%). The mean numbers of carcinomas per bladder in groups 1-5 were 1.2, 0.5, 0.4, 0.5 and 1.1, respectively. The incidence and the mean number of carcinomas per bladder were significantly lower in the treatment groups (P < 0.05) than in the control group, except in the low-dose meloxicam group. There were no significant adverse effects. CONCLUSION: Both meloxicam and gefitinib have inhibitory effects on rat bladder carcinogenesis with no significant adverse effects. A combination of these drugs would be worth studying for their synergistic effects. PMID- 16469041 TI - A comparison of p21 and p27 immunoexpression in benign glands, prostatic intraepithelial neoplasia and prostate adenocarcinoma. AB - OBJECTIVE: To assess the immunoexpression of p21 and p27 proteins in consecutive areas of benign glands, prostatic intraepithelial neoplasia (PIN) and prostate adenocarcinoma. PATIENTS AND METHODS: Tissue from 91 patients who had a radical prostatectomy was assessed by immunohistochemistry to evaluate the expression of p21 and p27 in adjacent areas of benign glands, PIN and prostate adenocarcinoma. The results were correlated with various clinicopathological variables, e.g. age, total prostate-specific antigen level, tumour stage, Gleason score, surgical margin involvement, extraprostatic extension, seminal vesicle invasion, and perineural invasion. RESULTS: The p27 score in PIN was lower than that in benign glands (P = 0.04) but there was no significant difference between the scores for PIN and tumour. By contrast, p21 expression was almost undetectable in benign glands, although there was substantially more in PIN and tumour (P < 0.01). Some patients had no expression of p21 in tumour tissue, and had lower p21 scores in benign glands and PIN areas (P < 0.05). Interestingly, these patients had a lower frequency of negative prognostic variables. The tumour p21 score was higher in patients with extraprostatic extension (P = 0.045) and tumour p27 expression was inversely correlated with seminal vesicle invasion (P = 0.028). Tumour and PIN p27 expression appeared to decrease with advancing age (P = 0.008 and 0.012, respectively). CONCLUSION: Higher p21 and lower p27 expression is correlated with adverse prognostic factors. The decline in p27 with advancing age in tumour and PIN areas may be a possible explanation of the greater frequency of prostate adenocarcinoma in older men. A p21-negative tumour subgroup with a lower frequency of adverse prognostic factors was identified, which needs to be further explored. PMID- 16469042 TI - What's the point? PMID- 16469045 TI - Diabetes mellitus is associated with subnormal levels of free testosterone in men. PMID- 16469046 TI - The need to reduce patient discomfort during transrectal ultrasonography-guided prostate biopsy: what do we know? PMID- 16469047 TI - Concomitant management of renal calculi and pelvi-ureteric junction obstruction with robotic laparoscopic surgery. PMID- 16469048 TI - Zoledronic acid is effective in preventing and delaying skeletal events in patients with bone metastases secondary to genitourinary cancers. PMID- 16469049 TI - Surgical management of penile carcinoma: the primary lesion. PMID- 16469050 TI - Interactions between DNA viruses, ND10 and the DNA damage response. AB - ND10 are small nuclear substructures that are defined by the presence the promyelocytic leukaemia protein PML. Many other proteins have been detected within ND10, a complexity that is reflected in reports of their involvement in multiple cellular pathways that include the regulation of gene expression, chromatin dynamics, protein modification, apoptosis, p53 function, senescence, DNA repair, the interferon response and viral infection. This review summarizes recent evidence of similarities between the behaviour of ND10 components and DNA repair pathway proteins in response to viral infection and DNA damage. ND10 structures become associated with the parental genomes and early replication compartments of many DNA viruses, and DNA repair pathway proteins are also recruited to these sites. Similarly, PML and DNA repair proteins are recruited to sites of DNA damage. The mechanisms by which these events might occur, and the implications for ND10 function in DNA virus infection and chromatin metabolism, are discussed. PMID- 16469051 TI - Ringing the alarm bells: signalling and apoptosis in influenza virus infected cells. AB - Small RNA viruses such as influenza viruses extensively manipulate host-cell functions to support their replication. At the same time the infected cell induces an array of defence mechanisms to fight the invader. These processes are mediated by a variety of intracellular signalling cascades. Here we will review the current knowledge of functional kinase signalling and apoptotic events in influenza virus infected cells and how these viruses have learned to misuse these cellular responses for efficient replication. PMID- 16469052 TI - Viral interactions with the cytoskeleton: a hitchhiker's guide to the cell. AB - The actin and microtubule cytoskeleton play important roles in the life cycle of every virus. During attachment, internalization, endocytosis, nuclear targeting, transcription, replication, transport of progeny subviral particles, assembly, exocytosis, or cell-to-cell spread, viruses make use of different cellular cues and signals to enlist the cytoskeleton for their mission. Viruses induce rearrangements of cytoskeletal filaments so that they can utilize them as tracks or shove them aside when they represent barriers. Viral particles recruit molecular motors in order to hitchhike rides to different subcellular sites which provide the proper molecular environment for uncoating, replicating and packaging viral genomes. Interactions between subviral components and cytoskeletal tracks also help to orchestrate virus assembly, release and efficient cell-to-cell spread. There is probably not a single virus that does not use cytoskeletal and motor functions in its life cycle. Being well informed intracellular passengers, viruses provide us with unique tools to decipher how a particular cargo recruits one or several motors, how these are activated or tuned down depending on transport needs, and how cargoes switch from actin tracks to microtubules to nuclear pores and back. PMID- 16469053 TI - Differential inductions of TNF-alpha and IGTP, IIGP by structurally diverse classic and non-classic lipopolysaccharides. AB - The innate immune system recognizes microbes by characteristic molecules like the Gram-negative lipopolysaccharide (LPS). Lipid A (the LPS bioactive moiety) signals through toll-like receptors (TLRs) to induce pro-inflammatory molecules and small GTPases of the p47 family involved in intracellular pathogen control. We tested TNF-alpha and p47-GTPase induction in macrophages using classical LPSs [lipid As with glucosamine backbones, ester- and amide-linked C14:0(3-OH) and C12 to C16 in acyloxyacyl groups] of wild type and mutant Escherichia coli and Yersinia species and non-classical LPSs [lipid As with diaminoglucose, ester linked 3-OH-fatty acids and C28:0(27-OH) and C23:0(29-OH) in acyloxyacyl groups] of plant endosymbionts (Rhizobium), intracellular pathogens (Brucella and Legionella) and phylogenetically related opportunistic bacteria (Ochrobactrum). Classical but not non-classical LPSs efficiently induced TNF-alpha, IIGP and IGTP p47-GTPase expression. Remarkably, the acyloxyacyl groups in classical LPSs necessary to efficiently induce TNF-alpha were not necessary to induce p47 GTPases, suggesting that different aspects of lipid A are involved in this differential induction. This was confirmed by using PPDM2, a non-endotoxic lipid A-structurally related synthetic glycolipid. Despite their different bioactivity, all types of LPSs signalled through TLR-4 and not through TLR-2. However, whereas TNF-alpha induction was myeloid differentiation factor 88 (MyD88)-dependent, that of p47-GTPases occurred via a MyD88-independent pathway. These observations show that different aspects of the LPS pathogen-associated molecular pattern may be triggering different signalling pathways linked to the same TLR. They also reinforce the hypothesis that non-classical lipid As act as virulence factors by favouring the escape from the innate immune system. PMID- 16469054 TI - TLR2 and TLR4 differentially regulate B7-1 resulting in distinct cytokine responses to the mycoplasma superantigen MAM as well as to disease induced by Mycoplasma arthritidis. AB - Mycoplasma arthritidis mitogen (MAM) is a superantigen secreted by M. arthritidis, an agent of murine arthritis and toxicity. We previously demonstrated that C3H mouse sub-strains differing in expression of Toll-like receptor 4 (TLR4), differed in immune reactivity to MAM due to differential engagement of TLR2 and TLR4. Here we examine the role of B7 co-stimulatory molecules in immune outcome and disease manifestations resulting from these different MAM/TLR2 and MAM/TLR4 interactions. Injections of MAM into C3H/HeJ mice upregulated expression of B7-1 but not B7-2 on peritoneal adherent cells, whereas B7-1 expression was lower on cells from C3H/HeSnJ mice. Anti-B7-1 antibody but not anti-B7-2, injected in vivo, changed the type 1 cytokines in MAM-injected C3H/HeJ mice to a type 2 cytokines and, conversely, the type 2 response in C3H/HeSnJ mice injected with anti-B7-1 shifted to a type 1 pattern. Whereas anti B7-2 exerted no effect on disease in either mouse strain, anti-B7-1 significantly delayed the lethal toxicity of M. arthritidis in C3H/HeJ mice but enhanced arthritis in C3H/HeSnJ mice. Thus, TLR-mediated regulation of B7-1 results in diverse cytokine profiles in C3H sub-strains, and that the interaction of MAM with different TLR(s) may differentially affect cytokine responses and ultimately, M. arthritidis disease. PMID- 16469055 TI - A vaccinia virus lacking A10L: viral core proteins accumulate on structures derived from the endoplasmic reticulum. AB - The assembly of the intracellular mature virus (IMV) of vaccinia virus (VV), the prototype member of the poxviridae, is poorly understood and controversial. We have previously proposed that the IMV is composed of a continuous double membraned cisterna derived from the smooth ER, whereby the genome-containing core is enwrapped by a part of this cisterna. In the present study we characterize a mutant virus in which the synthesis of the major core protein A10L can be conditionally expressed. Without A10L, IMVs are not made; immature viruses (IVs) and regularly stacked membrane structures that contain viral DNA, accumulate instead. By immunolabelling of thawed cryo-sections these stacks contain most of the viral core proteins and low levels of viral membrane proteins. Importantly, the stacked membranes could be labelled with antibodies to an ER marker protein, implying that they are derived from this cellular compartment. By electron tomography (ET) on semi-thin cryo-sections we show that the membranes of the stacks are continuous with the membranes of the IVs. Direct continuities with ER cisternae, to which the stacks are tightly apposed, were, however, not unequivocally seen. Finally, ET revealed how the IV membranes separated to become two-membrane profiles. Taken together, this study shows that VV core proteins and the viral DNA can coassemble onto ER-derived membranes that are continuous with the membranes of the IVs. PMID- 16469056 TI - Dictyostelium transcriptional host cell response upon infection with Legionella. AB - Differential gene expression of Dictyostelium discoideum after infection with Legionella pneumophila was investigated using DNA microarrays. Investigation of a 48 h time course of infection revealed several clusters of co-regulated genes, an enrichment of preferentially up- or downregulated genes in distinct functional categories and also showed that most of the transcriptional changes occurred 24 h after infection. A detailed analysis of the 24 h time point post infection was performed in comparison to three controls, uninfected cells and co-incubation with Legionella hackeliae and L. pneumophilaDeltadotA. One hundred and thirty-one differentially expressed D. discoideum genes were identified as common to all three experiments and are thought to be involved in the pathogenic response. Functional annotation of the differentially regulated genes revealed that apart from triggering a stress response Legionella apparently not only interferes with intracellular vesicle fusion and destination but also profoundly influences and exploits the metabolism of its host. For some of the identified genes, e.g. rtoA involvement in the host response has been demonstrated in a recent study, for others such a role appears plausible. The results provide the basis for a better understanding of the complex host-pathogen interactions and for further studies on the Dictyostelium response to Legionella infection. PMID- 16469057 TI - Species specificity of the Listeria monocytogenes InlB protein. AB - InlA and InlB mediate L. monocytogenes entry into eukaryotic cells. InlA is required for the crossing of the intestinal and placental barriers. InlA uses E cadherin as receptor in a species-specific manner. The human E-cadherin but not the mouse E-cadherin is a receptor for InlA. In human cells, InlB uses Met and gC1qR as receptors. By studying the role of InlB in vivo, we found that activation of Met by InlB is species-specific. In mice, InlB is important for liver and spleen colonization, but not for the crossing of the intestinal epithelium. Strikingly, the virulence of a DeltainlB deletion mutant is not attenuated in guinea pigs and rabbits. Guinea pig and rabbit cell lines do not respond to InlB, although expressing Met and gC1qR, but support InlB-mediated responses upon human Met gene transfection, indicating that InlB does not recognize or stimulate guinea pig and rabbit Met. In guinea pig cells, the effect of human Met gene transfection on InlB-dependent entry is increased upon cotransfection with human gc1qr gene, showing the additive roles of gC1qR and Met. These results unravel a second L. monocytogenes species specificity critical for understanding human listeriosis and emphasize the need for developing new animal models for studying InlA and InlB functions in the same animal model. PMID- 16469058 TI - The Crohn's disease-associated adherent-invasive Escherichia coli strain LF82 replicates in mature phagolysosomes within J774 macrophages. AB - Adherent-invasive Escherichia coli (AIEC) bacteria isolated from Crohn's disease patients are able to extensively replicate within macrophages in large vacuoles. The mechanism by which AIEC bacteria survive within phagocytic cells is unknown. This report describes the maturation of AIEC LF82-containing phagosomes within J774 macrophages. LF82-containing phagosomes traffic through the endocytic pathway as shown by the sequential acquisition and loss of EEA1 and Rab7 and by accumulation of Lamp-1, Lamp-2 and cathepsin D. We demonstrated that AIEC LF82 containing phagosomes mature into active phagolysosomes where bacteria are exposed to low pH and to the degradative activity of cathepsin D. Finally, we showed that an acidic environment is necessary for replication of AIEC LF82 bacteria within J774 macrophages. Thus, evidence is provided that AIEC LF82 bacteria do not escape from the endocytic pathway but undergo normal interaction with host endomembrane organelles and replicate within acidic and cathepsin D positive vacuolar phagolysosomes. PMID- 16469059 TI - Strong immunostimulatory activity of AT-oligodeoxynucleotide requires a six-base loop with a self-stabilized 5'-C...G-3' stem structure. AB - Lactobacillus gasseri OLL2716 has recently been discovered as a probiotic that suppresses the growth of Helicobacter pylori and reduces gastric mucosal inflammation in humans. This has resulted in the development of a new type of probiotic yoghurt 'LG21' in Japan. In our previous study, we found an immunostimulatory AT5ACL oligodeoxynucleotide (AT-ODN) containing a unique core sequence (5'-ATTTTTAC-3') in L. gasseri JCM1131(T). Interestingly, although the AT-ODN does not contain any CpG sequences, it exerts mitogenic activity in B cells and augments Th-1-type immune responses via Toll-like receptor 9. These findings prompted us to identify strong immunostimulatory non-CpG AT-ODNs that contain the 5'-ATTTTTAC-3' motif in the genomic sequence of L. gasseri OLL2716. We identified 280 kinds of AT-ODNs in the L. gasseri OLL2716 genome. Mitogenicity and NF-kappaB gene reporting assays showed that 13 of the 280 AT-ODNs were strongly immunostimulatory when in the TLR9 transfectant. Of these, AT-ODNs LGAT 145 and LGAT-243 were the most potent. With respect to the induction of Th-1-type cytokines, LGAT-243 had the greatest activity and was more potent than the swine prototype, ODN D25. We further found that a six-base secondary loop structure containing a self-stabilized 5'-C...G-3' stem sequence is important for potent immunostimulatory activity. These results show for the first time that AT-ODNs with a specific loop and stem structure are important factors for immunostimulatory activity. Finally, we found that novel strong immunostimulatory non-CpG AT-ODNs exist in the genome of probiotic lactic acid bacteria. PMID- 16469060 TI - Coxiella burnetii inhabits a cholesterol-rich vacuole and influences cellular cholesterol metabolism. AB - Coxiella burnetii directs the synthesis of a large parasitophorous vacuole (PV) required for replication. While some lysosomal characteristics of the PV have been described, the origin and composition of the PV membrane is largely undefined. Cholesterol is an essential component of mammalian cell membranes where it plays important regulatory and structural roles. Here we investigated the role of host cholesterol in biogenesis and maintenance of the C. burnetii PV in Vero cells. The C. burnetii PV membrane stained with filipin and was positive for the lipid raft protein flotillin-1, suggesting PV membranes are enriched in cholesterol and contain lipid raft microdomains. C. burnetii infection increased host cell cholesterol content by 1.75-fold with a coincident upregulation of host genes involved in cholesterol metabolism. Treatment with U18666A, lovastatin, or 25-hydroxycholesterol, pharmacological agents that inhibit cholesterol uptake and/or biosynthesis, altered PV morphology and partially inhibited C. burnetii replication. Complete inhibition of C. burnetii PV development and replication was observed when infected cells were treated with imipramine or ketoconazole, inhibitors of cholesterol uptake and biosynthesis respectively. We conclude that C. burnetii infection perturbs host cell cholesterol metabolism and that free access to host cholesterol stores is required for optimal C. burnetii replication. PMID- 16469061 TI - Co-ordinate but disproportionate activation of apoptotic, regenerative and inflammatory pathways characterizes the liver response to acute amebic infection. AB - The liver has the remarkable ability to respond to injury with repair and regeneration. The protozoan parasite Entamoeba histolytica is the major cause of liver abscess worldwide. We report a transcriptional analysis of the response of mouse liver to E. histolytica infection, the first study looking at acute liver infection by a non-viral pathogen. Focusing on early time points, we identified 764 genes with altered transcriptional levels in amebic liver abscess. The response to infection is rapid and complex, with concurrent increased expression of genes linked to host defence through IL-1, TLR2, or interferon-induced pathways, liver regeneration via activation of IL-6 pathways, and genes associated with programmed cell death possibly through TNFalpha or Fas pathways. A comparison of amebic liver infection with the liver response to partial hepatectomy or toxins reveals striking similarities between amebic liver abscess and non-infectious injury in key components of the liver regeneration pathways. However, the response in amebic liver abscess is biased towards apoptosis when compared with acute liver injury from hepatectomy, toxins, or other forms of liver infection. E. histolytica infection of the liver simultaneously activates inflammatory, regenerative and apoptotic pathways, but the sum of these early responses is biased towards programmed cell death. PMID- 16469062 TI - Differential expression of VirB9 and VirB6 during the life cycle of Anaplasma phagocytophilum in human leucocytes is associated with differential binding and avoidance of lysosome pathway. AB - Anaplasma phagocytophilum, an obligate intracellular bacterium, is the aetiologic agent of human granulocytic anaplasmosis (HGA). A. phagocytophilum virB/D operons encoding type IV secretion system are expressed in cell culture and in the blood of HGA patients. In the present study, their expression across the A. phagocytophilum intracellular developmental cycle was investigated. We found that mRNA levels of both virB9 and virB6 were upregulated during infection of human neutrophils in vitro. The antibody against the recombinant VirB9 protein was prepared and immunogold and immunofluorescence labelling were used to determine the VirB9 protein expression by individual organisms. Majority of A. phagocytophilum spontaneously released from the infected host cells poorly expressed VirB9. At 1 h post infection, VirB9 was not detectable on most bacteria associated with neutrophils. However, VirB9 was strongly expressed by A. phagocytophilum during proliferation in neutrophils. In contrast, with HL-60 cells, approximately 80% of A. phagocytophilum organisms associated at 1 h post infection expressed VirB9 protein and were colocalized with lysosome-associated membrane protein-1 (LAMP-1), whereas, VirB9-undetectable bacteria were not colocalized with LAMP-1. These results indicate developmental regulation of expression of components of type IV secretion system during A. phagocytophilum intracellular life cycle and suggest that bacterial developmental stages influence the nature of binding to the hosts and early avoidance of late endosome lysosome pathway. PMID- 16469063 TI - Genome-wide analysis of gene expression in adult Anopheles gambiae. AB - With their genome sequenced, Anopheles gambiae mosquitoes now serve as a powerful tool for basic research in comparative, evolutionary and developmental biology. The knowledge generated by these studies is expected to reveal molecular targets for novel vector control and pathogen transmission blocking strategies. Comparisons of gene-expression profiles between adult male and nonblood-fed female Anopheles gambiae mosquitoes revealed that roughly 22% of the genes showed sex-dependent regulation. Blood-fed females switch the majority of their metabolism to blood digestion and egg formation within 3 h after the meal is ingested, in detriment to other activities such as flight and response to environment stimuli. Changes in gene expression are most evident during the first, second and third days after a blood meal, when as many as 50% of all genes showed significant variation in transcript accumulation. After laying the first cluster of eggs (between 72 and 96 h after the blood meal), mosquitoes return to a nongonotrophic stage, similar but not identical to that of 3-day-old nonblood fed females. Ageing and/or the nutritional state of mosquitoes at 15 days after a blood meal is reflected by the down-regulation of approximately 5% of all genes. A full description of the large number of genes regulated at each analysed time point and each biochemical pathway or biological processes in which they are involved is not possible within the scope of this contribution. Therefore, we present descriptions of groups of genes displaying major differences in transcript accumulation during the adult mosquito life. However, a publicly available searchable database (http://www.angagepuci.bio.uci.edu/) has been made available so that detailed analyses of specific groups of genes based on their descriptions, functions or levels of gene expression variation can be performed by interested investigators according to their needs. PMID- 16469064 TI - Sequence variation in trypsin- and chymotrypsin-like cDNAs from the midgut of Ostrinia nubilalis: methods for allelic differentiation of candidate Bacillus thuringiensis resistance genes. AB - Midgut expressed alkaline serine proteases of Lepidoptera function in conversion of Bacillus thuringiensis (Bt) protoxin to active toxin, and reduced level of transcript T23 is associated with Ostrinia nubilalis resistance to Dipel Bt formulations. Three groups of trypsin- (OnT25, OnT23, and OnT3) and two chymotrypsin-like (OnC1 and OnC2) cDNAs were isolated from O. nubilalis midgut tissue. Intraspecific groupings are based on cDNA similarity and peptide phylogeny. Derived serine proteases showed a catalytic triad (His, Asp, and Ser; except transcript OnT23a), three substrate specificity-determining residues, and three paired disulphide bonds. RT-PCR indicated all transcripts are expressed in the midgut. Mendelian-inherited genomic markers for loci OnT23, OnT3 and OnC1 will be useful for association of alleles with bioassayed Bt toxin resistance phenotypes. PMID- 16469065 TI - Use of two transcription starts in the G6PD gene of the bark beetle Ips typographus. AB - The enzyme glucose-6-phosphate dehydrogenase (G6PD) of the bark beetle Ips typographus is derived from a gene that includes eight exons and spans over 7100 nucleotides (nt). By means of two transcription starts, the gene generates two mRNA isoforms that are present in similar amounts in the larvae, pupae and adults. The A isoform includes exon IA of 115 nt, which is followed by intron 1a extending to position 3457 of the gene. The B mRNA isoform begins with exon IB (100 nt) that occupies positions 3291-3390 within the 1a intron. Exons II to VII are included in both mRNA isoforms. The gene contains 31.6% (36.5% in the translated region) of the GC nucleotides. Two transcription starts and the exon/intron organization distinguish bark beetle G6PD from the homologous genes known in other insects. Two enzyme variants were detected in the protein extracts of individual bark beetles but their relationship to the A and B mRNA isoforms is uncertain. PMID- 16469067 TI - Evolution of spider silks: conservation and diversification of the C-terminus. AB - Analysis of DNA sequences coding for the C-terminus of spider silk proteins from a range of spiders suggests that many silk C-termini share a common origin, and that their physical properties have been highly conserved over several hundred million years. These physical properties are compatible with roles in protein synthesis, silk function and in recruiting accessory proteins. Phylogenetic relationships among different silk genes suggest that any recombination has been insufficient to homogenize the different types of silk gene, which appear to have evolved independently of one another. The types of nucleotide substitutions that have occurred suggest that selection may have operated differently in the various silk lineages. Amino acid sequences of flagelliform silk C-termini differ substantially from the other types of spider silk studied, but they are expected to have very similar physical properties and may perform a similar function. PMID- 16469066 TI - Gene expression profiles underlying alternative caste phenotypes in a highly eusocial bee, Melipona quadrifasciata. AB - To evaluate caste-biased gene expression in Melipona quadrifasciata, a stingless bee, we generated 1278 ESTs using Representational Difference Analysis. Most annotated sequences were similar to honey bee genes of unknown function. Only few queen-biased sequences had their putative function assigned by sequence comparison, contrasting with the worker-biased ESTs. The expression of six annotated genes connected to caste specificity was validated by real time PCR. Interestingly, queens that were developmentally induced by treatment with a juvenile hormone analogue displayed an expression profile clearly different from natural queens for this set of genes. In summary, this study represents an important first step in applying a comparative genomic approach to queen/worker polyphenism in the bee. PMID- 16469068 TI - Diptericin expression in bacteria infected Drosophila mbn-2 cells - effect of infection dose and phagocytosis. AB - Drosophila haemocytes play a key role in defence against microbial aggression. Their capacity to sense and dispose of bacteria and also to signal to other immune tissues is probably vital to overcome an infection. In this work we used the haemocyte-like mbn-2 cell line to investigate how expression of the antimicrobial peptide diptericin is affected after a high dose bacterial challenge with diaminopimelic acid (DAP)-peptidoglycan Gram-positive and Gram negative bacteria. We report that diptericin expression is negatively affected by high infection dose and rapid bacterial growth regardless of the type of infection and bacterial virulence and occurs in the absence of mbn-2 cell death. Furthermore we show that the mbn-2 cell population is heterogeneous, containing both phagocytic and nonphagocytic cells and that contact with large numbers of bacteria decreases diptericin expression in the phagocytic cell population. PMID- 16469069 TI - Extraordinary number of gene rearrangements in the mitochondrial genomes of lice (Phthiraptera: Insecta). AB - The arrangement of genes in the mitochondrial (mt) genomes of most insects is the same, or near-identical, to that inferred to be ancestral for insects. We sequenced the entire mt genome of the small pigeon louse, Campanulotes bidentatus compar, and part of the mt genomes of nine other species of lice. These species were from six families and the three main suborders of the order Phthiraptera. There was no variation in gene arrangement among species within a family but there was much variation in gene arrangement among the three suborders of lice. There has been an extraordinary number of gene rearrangements in the mitochondrial genomes of lice! PMID- 16469070 TI - A novel guanylyl cyclase receptor, BdmGC-1, is highly expressed during the development of the oriental fruit fly Bactrocera dorsalis (Hendel). AB - A novel receptor guanylyl cyclase (GC) has been identified from the oriental fruit fly Bactrocera dorsalis (Hendel) and has been designated BdmGC-1. Protein domain analysis revealed that BdmGC-1 possesses a characteristic domain organization similar to all known receptor GCs but with a unique carboxyl terminal extension. When overexpressed in 293T cells, BdmGC-1 manifests as a cell surface glycoprotein with a marked cGMP-generating activity but is unresponsive to all ligands known to activate mammalian receptor GCs. BdmGC-1 mRNAs were highly expressed during development but had low or no expression in adult tissues. On the basis of its unique sequence and distinct developmental expression pattern, BdmGC-1 represents a novel receptor GC that may play a critical role during the development of B. dorsalis. PMID- 16469071 TI - Receptor of activated C kinase 1 (RACK1) is necessary for the 20-hydroxyecdysone induced expression of the transcription factor CHR3 in the spruce budworm Choristoneura fumiferana. AB - To initiate moulting and metamorphosis, 20-hydroxyecdysone (20E) binds to its nuclear receptors and the ligand-receptor complex then mediates changes in gene expression. Phosphorylation of the receptors is required for their function. The intracellular signal transduction pathway that is involved in receptor phosphorylation remains elusive. This study provides evidence that the receptor of activated C kinase 1 (RACK1) and protein kinase C (PKC) signal transduction cascade is involved in the 20E-induced expression of the moult-associated transcription factor CHR3. A cDNA clone encoding a receptor of activated C kinase 1 was isolated from Choristoneura fumiferana (CfRACK1). This single copy gene coded a 36 kDa protein and was expressed ubiquitously in all of the developmental stages and the tissues tested, including the midgut, epidermis, fat body, head, Malpighian tubules, ovary and testis of larvae. High levels of the transcripts were also detected in a midgut-derived CF-203 cell line. We noticed that the green fluorescence protein-fused CfRACK1 protein was distributed in the cytosol surrounding the nuclei in stably transformed cells. Interference of CfRACK1 mRNA suppressed the 20E-induced expression of the transcription factor CHR3. Dequalinium-14; 1,1'-decamethylenebis-4-aminoquinaldinium diiodide (DECA), an inhibitor of RACK1 binding to protein kinase C, blocked the 20E-induced expression of CHR3 and accumulation of the ecdysone receptor (EcR) in the nuclei. All of these data together suggest that 20E-induced expression of CHR3 may involve phosphorylation of the ecdysone receptor component through the PKC/RACK1 signal transduction cascade, which facilitates the import of the receptor into the nuclei of cells. PMID- 16469072 TI - Distribution and prevalence of Wolbachia in introduced populations of the fire ant Solenopsis invicta. AB - Wolbachia are intracellular bacteria that induce phenotypic effects in many arthropod hosts to enhance their own transmission within host populations. Wolbachia commonly infect the Red Imported Fire Ant, Solenopsis invicta, in native South American populations. A previous study failed to detect Wolbachia in fire ants from the introduced range in the USA. We conducted an extensive study of individuals collected from 1157 nests from 10 widespread geographical populations in the USA. Wolbachia were detected in ants from two nearby populations in southern Mississippi, with different variants (wsp gene sequences) infecting ants from colonies of the multiple-queen (polygyne) vs. single-queen (monogyne) social forms. The parsimonious explanation for the presence of Wolbachia in introduced S. invicta is that there have been one or more recent introductions of Wolbachia-infected fire ants into the southern USA. PMID- 16469073 TI - Germ line transformation of the olive fly Bactrocera oleae using a versatile transgenesis marker. AB - The olive fruit fly (olive fly) Bactrocera oleae (Dacus), recently introduced in North America, is the most destructive pest of olives worldwide. The lack of an efficient gene transfer technology for olive fly has hampered molecular analysis, as well as development of genetic techniques for its control. We have developed a Minos-based transposon vector carrying a self-activating cassette which overexpresses the enhanced green fluorescent protein (EGFP). Efficient transposase-mediated integration of one to multiple copies of this vector was achieved in the germ line of B. oleae by coinjecting the vector along with in vitro synthesized Minos transposase mRNA into preblastoderm embryos. The self activating gene construct combined with transposase mRNA present a system with potential for transgenesis of very diverse species. PMID- 16469077 TI - Cytokeratin 15 expression in apocrine mixed tumors of the skin and other benign neoplasms with apocrine differentiation. AB - To clarify the features of apocrine mixed tumors (AMT) of the skin among benign neoplasms with apocrine differentiation in their relationship to follicular stem cells, we investigated the immunohistochemical expression of CK15 (LHK15 and C8/144B), which is a relatively specific marker of hair follicle stem cells in the bulge, in 35 cases of eight different benign neoplasms with presumed apocrine differentiation. All eight cases of AMT of the skin showed CK15 immunostaining of the neoplastic cells, and all four cases of syringocystadenoma papilliferum, all five cases of spiradenoma, and both cases of cylindroma also showed a focally positive reaction to CK15. None of the other benign neoplasms with presumed apocrine differentiation showed CK15 expression. In AMT of the skin, the proportion of CK15-positive cells in the follicular or sebaceous differentiation group (78.8%, average of four cases) was significantly higher than the group without this differentiation (8.8%, average of four cases). AMT of the skin are unique among benign neoplasms with apocrine differentiation in their substantial and constant CK15 expression, suggesting that they derive from multipotent epithelial stem cells in the bulge. AMT of the skin with follicular or sebaceous differentiation are considered to show an immature stage of apocrine differentiation still rich in stem cells or to originate from stem cells with an incompletely established apocrine fate. The partially positive reaction for CK15 in syringocystadenomas papilliferum and spiradenoma/cylindroma may depend on the ability to express CK15 in stem cells with an apocrine fate or result from the follicular and apocrine nature of this neoplasm. PMID- 16469078 TI - Temperament and character dimensions in patients with atopic dermatitis. AB - The aim of this study was to assess the personality traits of young male patients with atopic dermatitis (AD), and to examine the correlations between temperament and character dimensions with clinical and other psychological factors. Fifty young adult male AD patients and 83 healthy controls were examined using the temperament and character inventory, the Beck depression inventory and the state trait anxiety inventory. The AD patients scored higher on harm avoidance and lower on reward dependence, self-directedness and cooperativeness than the healthy controls. The illness duration and anxiety correlated negatively with the self-directedness score, and depression correlated negatively with reward dependence and the persistence scores in AD patients. These results suggest that AD patients have distinctive temperament and character dimensions compared to healthy controls. Moreover, illness duration and anxiety might be associated with some personality problems, and some temperament dimensions (e.g. reward dependence, persistence) may be linked to depressive symptoms in AD patients. PMID- 16469079 TI - Effects of alpha-hydroxy acids on the human skin of Japanese subjects: the rationale for chemical peeling. AB - Alpha-hydroxy acid (AHA) agents, such as glycolic acid and lactic acid, have been used as therapeutic agents for more than a quarter of a century. Recently, they have been used as agents to rejuvenate photo-aged skin. It is believed that these AHA agents induce the epidermis to remodel and accelerate desquamation, thus exerting their therapeutic effects. In this study, we investigated the histological differences in skin treated with glycolic, lactic, citric and acetic acids once daily for 6 weeks. The melanin pigments in the basal layer were less prominent in the glycolic and lactic acid-treated skin than in the citric and acetic acid-treated skin. The melanin deposits in the horny layers were equal for all AHA. However, the melanin deposits in the squamous layers were less prominent in the glycolic and lactic acid-treated skins than in the citric and acetic acid treated skins; this was analogous to observations of the basal layers. Collagen I and procollagen I were increased after treatment with glycolic, lactic and citric acid in the upper dermis, but were not increased with acetic acid treatment. However, the staining of the epidermis and dermis for matrix metalloproteinase-1 (MMP-1) after treatment was not significantly different among the agents. Our data suggest that longer treatment intervals with glycolic and lactic acid can cause improvements in both the epidermal and dermal components and support the usefulness of AHA for rejuvenating photo-damaged skin. PMID- 16469080 TI - A case of tinea nigra palmaris in Okinawa, Japan. AB - We report a case of tinea nigra on the left palm of a 13-year-old girl. She had noticed a pigmented, asymptomatic macule on the left palm approximately 4-5 years prior to her first visit to our hospital. The color of the lesion tended to change before and after a bath; it became lighter after a bath and darkened some time later. Physical examination revealed that the macule was 4 cm x 5 cm in size, dark brown in color and irregularly shaped. Direct potassium hydroxide (KOH) microscopic examination from skin scrapings revealed branched brown hyphae with light brown septa. A fungal culture on Sabouraud's agar media produced wet, medium brown, yeast-like colonies, the surface of which later became black and shiny. A slide culture disclosed light brown, elliptic or peanut-shaped conidia comprised of one to two ampullaceous cells. Scanning electron microscopic examination of the conidia showed both annellation conidia with lunate bud scars and sympodial conidiogenesis. Using extracted DNA from separately cultured fungi, we performed polymerase chain reaction with the primers specific to Hortaea werneckii. The results showed positive bands. We performed direct sequencing with the DNA segments from the positive bands. The causative fungus in our case was determined to be type C of H. werneckii on the grounds of the base sequences obtained. The final diagnosis of the present case was made as tinea nigra by H. werneckii. We also report a brief survey of all the cases of tinea nigra reported in Japan to date. PMID- 16469081 TI - Adult variant of self-healing papular mucinosis in a patient with rheumatoid arthritis: predominant proliferation of dermal dendritic cells expressing CD34 or factor XIIIa in association with dermal deposition of mucin. AB - According to a recent classification, self-healing papular mucinosis (SHPM) is a subtype of papular mucinosis (also known as lichen myxedematosus), which is in turn a type of idiopathic localized cutaneous mucinosis. SHPM tends to occur in children, but there have been a few reports of an adult type. We report a 70-year old Japanese woman who presented with reddish, rice-kernel-sized papules of a few days' duration on her right arm. She had a 25-year history of rheumatoid arthritis, which had been well treated with a low dose of corticosteroid as well as some other medications. No paraproteinemia or thyroid dysfunction were observed. The eruptions spontaneously resolved within 2.5 months of onset. Histological findings showed a well-circumscribed mucinous stroma surrounded by dermal mesenchymal cells, such as fibroblast-like cells in the middle of the dermis. Immunohistochemically, these cells were positive only for vimentin on the mucinous lesion. On the circumference of the mucinous lesion, these cells expressed either CD34 or factor XIIIa (FXIIIa). Because vimentin was common to dermal mesenchymal cells, we defined the cells expressing CD34 or FXIIIa, except for vimentin+ cells lacking CD34 or FXIIIa, as dermal dendritic cells (DDC). The findings of the present case suggest that CD34+ or FXIIIa+ DDC and tryptase positive mast cells on the perilesional area in combination with vimentin+ cells on the mucinous lesion might have given rise to the dermal deposition of mucin in our case. These cells, which are possibly activated in an autoimmune manner associated with rheumatoid arthritis, might play important roles in the development of dermal deposition of mucin in SHPM. PMID- 16469082 TI - Syringoma-like eccrine sweat duct proliferation induced by radiation. AB - We report a 45-year-old woman with breast cancer who had undergone surgery and radiation and anti-estrogen therapy and presented with many reddish papules in the irradiated breast area. Skin biopsy of the affected area disclosed proliferation of eccrine sweat ducts and cystic structures; the clinical and histopathological features were consistent with syringoma-like eccrine sweat duct proliferation. The lesions spread rapidly on her chest during radiation therapy and regressed spontaneously 3 weeks after its completion. We postulate that the lesions were induced by radiation, and promoted by anti-estrogen therapy. PMID- 16469083 TI - A case of bullous dermatitis artefacta possibly induced by a deodorant spray. AB - Dermatitis artefacta is one of a spectrum of factitious diseases etiologically responsible for skin lesions denied by patients. These factors often make it difficult to identify the causative agents of the condition. Herein, we report a case of bullous dermatitis artefacta in a 12-year-old girl, for which a deodorant spray was suspected as the probable cause. Pathological examination revealed subepidermal blistering with full-thickness necrosis of the epidermis, suggesting a thermo- or cryo-induced injury. Psychological testing demonstrated her immaturity and dependence. In searching for the causative agent, we suspected a deodorant spray as a blister-inducing agent. We succeeded in reproducing a similar blister lesion on the volunteer's healthy skin using the same spray. Psychiatric involvement significantly complicates the treatment of factitious diseases, including dermatitis artefacta. Cooperation among dermatologists, psychiatrists and the patient's family members is required for ensuring a favorable prognosis. PMID- 16469084 TI - Granulomas induced by subcutaneous injection of leuprorelin acetate. AB - Leuprorelin acetate, a chemotherapeutic agent used to treat prostate cancer, is a synthetic luteinizing hormone-releasing hormone (LHRH) agonist. We report a 75 year-old man who presented with several large subcutaneous nodules at the site of leuprorelin acetate injections for his prostatic cancer. A biopsy of the nodules disclosed epithelioid granulomatous inflammation and resulted in a diagnosis of drug-induced granulomatous reaction to leuprorelin acetate. PMID- 16469085 TI - A case of Sister Mary Joseph's nodule from prostatic cancer. AB - The metastasis of a visceral malignancy to the umbilicus is known as "Sister Mary Joseph's nodule" (SMJN). We recently encountered an 80-year-old man in whom umbilical metastasis of prostatic cancer was revealed during the course of treatment. The lesion was a red, firm, elastic nodule, 2.5 cm in diameter, with a bumpy, irregular surface, but without subjective symptoms. In histopathological examination, atypical cells with intensely stained, small quasi-circular nuclei and weakly eosinophilic cytoplasm were found in all layers of the dermis. In some parts of these tumor cells, a gland-like structure was observed. Immunohistochemical investigation of the invading tumor cells disclosed immunoreactivity for prostate specific antigen and prostatic acid phosphatase, supporting a diagnosis of SMJN from prostatic cancer. His general condition gradually deteriorated, and he died 2 months later. Prostate cancer often metastasizes to bones and lymph nodes, but rarely to the skin. Of all the cases of cutaneous metastasis of prostate cancer reported in Japan, only three metastases have occurred to the umbilical region. The present case is therefore an example of a very rare condition. PMID- 16469086 TI - Acquired ichthyosis associated with an overlap syndrome of systemic sclerosis and systemic lupus erythematosus. AB - Acquired ichthyosis is a condition accompanying many systemic illnesses such as lymphoma, sarcoidosis, dermatomyositis and systemic lupus erythematosus (SLE). Overlap syndromes are defined as clinical entities which satisfy each of the diagnostic criteria of two different connective tissue diseases concurrently or consecutively. The coexistence of SLE with systemic sclerosis has been very rarely reported. We describe a 33-year-old woman with an overlap syndrome consisting of systemic sclerosis and SLE who developed ichthyosis on her extremities. PMID- 16469087 TI - Adult-onset Still's disease with prurigo pigmentosa-like skin eruption. AB - A 34-year-old woman with adult-onset Still's disease (AOSD) developed prurigo pigmentosa-like lesions on her chest and upper back in addition to the typical rash of AOSD. A biopsy specimen taken from the upper back showed characteristic features of prurigo pigmentosa. The eruption and fever subsided immediately after the administration of 40 mg/day prednisolone, but arthralgia persisted even after intravenous pulse methylprednisolone therapy in combination with immunosuppressive drugs. Various atypical skin rashes, including prurigo pigmentosa-like lesions, have been reported in association with AOSD. Therefore, one should carefully follow the clinical course of a patient in order not to overlook these atypical cutaneous manifestations of AOSD. PMID- 16469088 TI - A case of Papillon-Lefevre syndrome associated with xanthogranulomatous pyelonephritis and hepatitis. AB - Papillon-Lefevre syndrome (PLS) is an autosomally recessive palmoplantar keratoderma accompanied by psoriasiform plaques on the extensor surfaces of extremities and leading to premature loss of deciduous and permanent teeth by progressive periodontitis. Patients with PLS may exhibit mental retardation, intracranial ectopic calcifications, nail dystrophies and a tendency to various infectious disorders, in addition to skin and oral findings. Herein, we report a 26-year-old man with PLS, who had experienced xanthogranulomatous pyelonephritis and hepatitis during childhood and adolescence. To the best of our knowledge, this is the first report of PLS associated with xanthogranulomatous inflammation. PMID- 16469089 TI - A case of diffuse plane normolipemic xanthomatosis associated with pancytopenia and monoclonal gammopathy. AB - We report a case of diffuse plane normolipemic xanthomatosis (DPNX) which showed poorly demarcated, uncommon, yellow macules symmetrically distributed on the nape, axillae and inguinal folds accompanied by severe, persistent itching. Histopathological and ultrastructural studies of skin biopsy specimens revealed the existence of some foamy cells and the deposition of neutral fat in the upper papillary dermis. Laboratory investigations and bone marrow aspirate smears showed that our patient had myelodysplastic syndrome (MDS) associated with pancytopenia and monoclonal gammopathy of undetermined significance. Because our patient had neither a malignant hematological disorder nor a severe systemic disease, monoclonal gammopathy might explain the pathogenesis of DPNX in the present case. PMID- 16469090 TI - Peristomal pyoderma gangrenosum associated with rectal adenocarcinoma. PMID- 16469091 TI - A case of confluent and reticulated papillomatosis that successfully responded to roxithromycin. PMID- 16469092 TI - Subcutaneous epithelioid angiomatous nodule: a variant of epithelioid hemangioma. PMID- 16469093 TI - Signs of positive selection of somatic mutations in human cancers detected by EST sequence analysis. AB - BACKGROUND: Carcinogenesis typically involves multiple somatic mutations in caretaker (DNA repair) and gatekeeper (tumor suppressors and oncogenes) genes. Analysis of mutation spectra of the tumor suppressor that is most commonly mutated in human cancers, p53, unexpectedly suggested that somatic evolution of the p53 gene during tumorigenesis is dominated by positive selection for gain of function. This conclusion is supported by accumulating experimental evidence of evolution of new functions of p53 in tumors. These findings prompted a genome wide analysis of possible positive selection during tumor evolution. METHODS: A comprehensive analysis of probable somatic mutations in the sequences of Expressed Sequence Tags (ESTs) from malignant tumors and normal tissues was performed in order to access the prevalence of positive selection in cancer evolution. For each EST, the numbers of synonymous and non-synonymous substitutions were calculated. In order to identify genes with a signature of positive selection in cancers, these numbers were compared to: i) expected numbers and ii) the numbers for the respective genes in the ESTs from normal tissues. RESULTS: We identified 112 genes with a signature of positive selection in cancers, i.e., a significantly elevated ratio of non-synonymous to synonymous substitutions, in tumors as compared to 37 such genes in an approximately equal sized EST collection from normal tissues. A substantial fraction of the tumor specific positive-selection candidates have experimentally demonstrated or strongly predicted links to cancer. CONCLUSION: The results of EST analysis should be interpreted with extreme caution given the noise introduced by sequencing errors and undetected polymorphisms. Furthermore, an inherent limitation of EST analysis is that multiple mutations amenable to statistical analysis can be detected only in relatively highly expressed genes. Nevertheless, the present results suggest that positive selection might affect a substantial number of genes during tumorigenic somatic evolution. PMID- 16469094 TI - The war on marijuana: the transformation of the war on drugs in the 1990s. AB - BACKGROUND: As the "war on drugs" enters the latter half of its third decade since being forged into the American lexicon by President Ronald Reagan, the public has grown more skeptical of the current strategy and has proven to be receptive to a broader consideration of alternatives to incarceration. This has been the case most notably with marijuana offenses, where the policy discussion has shifted in some localities to one of decriminalization or de-prioritizing law enforcement resources dedicated to pursuing possession offenses. Despite the increased profile surrounding marijuana policy in recent years, there remains a significant degree of misunderstanding regarding the current strategy, both in terms of how resources are being allocated and to what eventual gain. METHODS: Previous studies have analyzed drug offenses as a general category, but there has yet to be a single study that has focused specifically on marijuana offenders at all stages of the system. This report analyzes multiple sources of data for the period 1990-2002 from each of the critical points in the criminal justice system, from arrest through court processing and into the correctional system, to create an overall portrait of this country's strategy in dealing with marijuana use. RESULTS: The study found that since 1990, the primary focus of the war on drugs has shifted to low-level marijuana offenses. During the study period, 82% of the increase in drug arrests nationally (450,000) was for marijuana offenses, and virtually all of that increase was in possession offenses. Of the nearly 700,000 arrests in 2002, 88% were for possession. Only 1 in 18 of these arrests results in a felony conviction, with the rest either being dismissed or adjudicated as a misdemeanor, meaning that a substantial amount of resources, roughly 4 billion dollars per year for marijuana alone, is being dedicated to minor offenses. CONCLUSION: The results of this study suggest that law enforcement resources are not being effectively allocated to offenses which are most costly to society. The financial and personnel investment in marijuana offenses, at all points in the criminal justice system, diverts funds away from other crime types, thereby representing a questionable policy choice. PMID- 16469095 TI - Geostatistical analysis of disease data: visualization and propagation of spatial uncertainty in cancer mortality risk using Poisson kriging and p-field simulation. AB - BACKGROUND: Smoothing methods have been developed to improve the reliability of risk cancer estimates from sparsely populated geographical entities. Filtering local details of the spatial variation of the risk leads however to the detection of larger clusters of low or high cancer risk while most spatial outliers are filtered out. Static maps of risk estimates and the associated prediction variance also fail to depict the uncertainty attached to the spatial distribution of risk values and does not allow its propagation through local cluster analysis. This paper presents a geostatistical methodology to generate multiple realizations of the spatial distribution of risk values. These maps are then fed into spatial operators, such as in local cluster analysis, allowing one to assess how risk spatial uncertainty translates into uncertainty about the location of spatial clusters and outliers. This novel approach is applied to age-adjusted breast and pancreatic cancer mortality rates recorded for white females in 295 US counties of the Northeast (1970-1994). A public-domain executable with example datasets is provided. RESULTS: Geostatistical simulation generates risk maps that are more variable than the smooth risk map estimated by Poisson kriging and reproduce better the spatial pattern captured by the risk semivariogram model. Local cluster analysis of the set of simulated risk maps leads to a clear visualization of the lower reliability of the classification obtained for pancreatic cancer versus breast cancer: only a few counties in the large cluster of low risk detected in West Virginia and Southern Pennsylvania are significant over 90% of all simulations. On the other hand, the cluster of high breast cancer mortality in Niagara county, detected after application of Poisson kriging, appears on 60% of simulated risk maps. Sensitivity analysis shows that 500 realizations are needed to achieve a stable classification for pancreatic cancer, while convergence is reached for less than 300 realizations for breast cancer. CONCLUSION: The approach presented in this paper enables researchers to generate a set of simulated risk maps that are more realistic than a single map of smoothed mortality rates and allow the propagation of cancer risk uncertainty through local cluster analysis. Coupled with visualization and querying capabilities of geographical information systems, animated display of realizations can highlight areas that depart consistently from the general behavior observed across the region, guiding further investigation and control activities. PMID- 16469096 TI - Relation between air pollution and allergic rhinitis in Taiwanese schoolchildren. AB - BACKGROUND: Recent findings suggest that exposure to outdoor air pollutants may increase the risk of allergic rhinitis. The results of these studies are inconsistent, but warrant further attention. The objective of the study was to assess the effect of relation between exposure to urban air pollution and the prevalence allergic rhinitis among school children. METHODS: We conducted a nationwide cross-sectional study of 32,143 Taiwanese school children. We obtained routine air-pollution monitoring data for sulphur dioxide (SO2), nitrogen oxides (NOx), ozone (O3), carbon monoxide (CO), and particles with an aerodynamic diameter of 10 microm or less (PM10). A parent-administered questionnaire provided information on individual characteristics and indoor environments (response rate 92%). Municipal-level exposure was calculated using the mean of the 2000 monthly averages. The effect estimates were presented as odds ratios (ORs) per 10 ppb change for SO2, NOx, and O3, 100 ppb change for CO, and 10 microg/m3 change for PM10. RESULTS: In two-stage hierarchical model adjusting for confounding, the prevalence of allergic rhinitis was significantly associated with SO2 (adjusted odds ratio (OR) = 1.43, 95% confidence interval (CI): 1.25, 1.64), CO (aOR = 1.05, 95% CI: 1.04, 1.07), and NOx (aOR = 1.11, 95% CI: 1.08, 1.15). Contrary to our hypothesis, the prevalence of allergic rhinitis was weakly or not related to O3 (aOR = 1.05, 95% CI: 0.98, 1.12) and PM10 (aOR = 1.00, 95% CI: 0.99, 1.02). CONCLUSION: Persistent exposure to NOx, CO, and SO2 may increase the prevalence of allergic rhinitis in children. PMID- 16469097 TI - HotSwap for bioinformatics: a STRAP tutorial. AB - BACKGROUND: Bioinformatics applications are now routinely used to analyze large amounts of data. Application development often requires many cycles of optimization, compiling, and testing. Repeatedly loading large datasets can significantly slow down the development process. We have incorporated HotSwap functionality into the protein workbench STRAP, allowing developers to create plugins using the Java HotSwap technique. RESULTS: Users can load multiple protein sequences or structures into the main STRAP user interface, and simultaneously develop plugins using an editor of their choice such as Emacs. Saving changes to the Java file causes STRAP to recompile the plugin and automatically update its user interface without requiring recompilation of STRAP or reloading of protein data. This article presents a tutorial on how to develop HotSwap plugins. STRAP is available at http://strapjava.de and http://www.charite.de/bioinf/strap. CONCLUSION: HotSwap is a useful and time saving technique for bioinformatics developers. HotSwap can be used to efficiently develop bioinformatics applications that require loading large amounts of data into memory. PMID- 16469098 TI - Gene prediction in eukaryotes with a generalized hidden Markov model that uses hints from external sources. AB - BACKGROUND: In order to improve gene prediction, extrinsic evidence on the gene structure can be collected from various sources of information such as genome genome comparisons and EST and protein alignments. However, such evidence is often incomplete and usually uncertain. The extrinsic evidence is usually not sufficient to recover the complete gene structure of all genes completely and the available evidence is often unreliable. Therefore extrinsic evidence is most valuable when it is balanced with sequence-intrinsic evidence. RESULTS: We present a fairly general method for integration of external information. Our method is based on the evaluation of hints to potentially protein-coding regions by means of a Generalized Hidden Markov Model (GHMM) that takes both intrinsic and extrinsic information into account. We used this method to extend the ab initio gene prediction program AUGUSTUS to a versatile tool that we call AUGUSTUS+. In this study, we focus on hints derived from matches to an EST or protein database, but our approach can be used to include arbitrary user-defined hints. Our method is only moderately effected by the length of a database match. Further, it exploits the information that can be derived from the absence of such matches. As a special case, AUGUSTUS+ can predict genes under user-defined constraints, e.g. if the positions of certain exons are known. With hints from EST and protein databases, our new approach was able to predict 89% of the exons in human chromosome 22 correctly. CONCLUSION: Sensitive probabilistic modeling of extrinsic evidence such as sequence database matches can increase gene prediction accuracy. When a match of a sequence interval to an EST or protein sequence is used it should be treated as compound information rather than as information about individual positions. PMID- 16469099 TI - Inflammation and enhanced nociceptive responses to bladder distension produced by intravesical zymosan in the rat. AB - BACKGROUND: Mycotic infections of the bladder produce pain and inflammatory changes. The present study examined the inflammatory and nociceptive effects of the yeast cell wall component, zymosan, when administered into the urinary bladder in order to characterize this form of bladder sensitization. METHODS: Parametric analyses of the time-course (0-48 hr) and concentration (0-2% solutions) variables associated with intravesical zymosan-induced bladder inflammation were performed in female rats. Plasma extravasation of Evan's Blue dye was used as a measure of tissue inflammation. Cardiovascular and visceromotor responses to urinary bladder distension were used as measures of nociception. RESULTS: Zymosan-induced bladder inflammation, as indexed by plasma extravasation of Evan's Blue, was significantly greater in rats treated with either 1 or 2% solutions as compared to either 0.1 or 0.5% zymosan solutions. In time-course studies (1-48 hr post-treatment), 1% zymosan-induced inflammation progressively increased with time following administration, was greatest at 24 hr and began to normalize by 48 hr. In the studies of inflammation-induced changes in nociception, arterial blood pressure (ABP) and visceromotor responses to graded distension of the urinary bladder were significantly increased relative to controls 24 hr after zymosan administration. CONCLUSION: These studies provide important time-course and solution concentration parameters for studies of zymosan-induced inflammation of the bladder and suggest utility of this model for the study of bladder-related pain. PMID- 16469100 TI - The association between processes, structures and outcomes of secondary prevention care among VA ischemic heart disease patients. AB - BACKGROUND: Hyperlipidemia and hypertension are well-established risk factors for recurrent cardiovascular events among patients with ischemic heart disease (IHD). Despite national recommendations, concordance with guidelines for LDL cholesterol and blood pressure remains inadequate. The objectives of this study were to 1) determine concordance rates with LDL cholesterol and BP recommendations; and 2) identify patient factors, processes and structures of care associated with guideline concordance among VA IHD patients. METHODS: This was a cross sectional study of veterans with IHD from 8 VA hospitals. Outcomes were concordance with LDL guideline recommendations (LDL<100 mg/dl), and BP recommendations (<140/90 mm Hg). Cumulative logit and hierarchical logistic regression analyses were performed to identify patient factors, processes, and structures of care independently associated with guideline concordance. RESULTS: Of 14,114 veterans with IHD, 55.7% had hypertension, 71.5% had hyperlipidemia, and 41.6% had both conditions. Guideline concordance for LDL and BP were 38.9% and 53.4%, respectively. However, only 21.9% of the patients achieved both LDL <100 mg/dl and BP <140/90 mm Hg. In multivariable analyses, patient factors including older age and the presence of vascular disease were associated with worse guideline concordance. In contrast, diabetes was associated with better guideline concordance. Several process of care variables, including higher number of outpatient visits, higher number of prescribed medications, and a recent cardiac hospitalization were associated with better guideline concordance. Among structures of care, having on-site cardiology was associated with a trend towards better guideline concordance. CONCLUSION: Guideline concordance with secondary prevention measures among IHD patients remains suboptimal. It is hoped that the findings of this study can serve as an impetus for quality improvement efforts to improve upon secondary prevention measures and reduce the morbidity and mortality of patients with known IHD. PMID- 16469101 TI - Evolution of the NANOG pseudogene family in the human and chimpanzee genomes. AB - BACKGROUND: The NANOG gene is expressed in mammalian embryonic stem cells where it maintains cellular pluripotency. An unusually large family of pseudogenes arose from it with one unprocessed and ten processed pseudogenes in the human genome. This article compares the NANOG gene and its pseudogenes in the human and chimpanzee genomes and derives an evolutionary history of this pseudogene family. RESULTS: The NANOG gene and all pseudogenes except NANOGP8 are present at their expected orthologous chromosomal positions in the chimpanzee genome when compared to the human genome, indicating that their origins predate the human-chimpanzee divergence. Analysis of flanking DNA sequences demonstrates that NANOGP8 is absent from the chimpanzee genome. CONCLUSION: Based on the most parsimonious ordering of inferred source-gene mutations, the deduced evolutionary origins for the NANOG pseudogene family in the human and chimpanzee genomes, in order of most ancient to most recent, are NANOGP6, NANOGP5, NANOGP3, NANOGP10, NANOGP2, NANOGP9, NANOGP7, NANOGP1, and NANOGP4. All of these pseudogenes were fixed in the genome of the human-chimpanzee common ancestor. NANOGP8 is the most recent pseudogene and it originated exclusively in the human lineage after the human chimpanzee divergence. NANOGP1 is apparently an unprocessed pseudogene. Comparison of its sequence to the functional NANOG gene's reading frame suggests that this apparent pseudogene remained functional after duplication and, therefore, was subject to selection-driven conservation of its reading frame, and that it may retain some functionality or that its loss of function may be evolutionarily recent. PMID- 16469103 TI - The DtxR protein acting as dual transcriptional regulator directs a global regulatory network involved in iron metabolism of Corynebacterium glutamicum. AB - BACKGROUND: The knowledge about complete bacterial genome sequences opens the way to reconstruct the qualitative topology and global connectivity of transcriptional regulatory networks. Since iron is essential for a variety of cellular processes but also poses problems in biological systems due to its high toxicity, bacteria have evolved complex transcriptional regulatory networks to achieve an effective iron homeostasis. Here, we apply a combination of transcriptomics, bioinformatics, in vitro assays, and comparative genomics to decipher the regulatory network of the iron-dependent transcriptional regulator DtxR of Corynebacterium glutamicum. RESULTS: A deletion of the dtxR gene of C. glutamicum ATCC 13032 led to the mutant strain C. glutamicum IB2103 that was able to grow in minimal medium only under low-iron conditions. By performing genome wide DNA microarray hybridizations, differentially expressed genes involved in iron metabolism of C. glutamicum were detected in the dtxR mutant. Bioinformatics analysis of the genome sequence identified a common 19-bp motif within the upstream region of 31 genes, whose differential expression in C. glutamicum IB2103 was verified by real-time reverse transcription PCR. Binding of a His tagged DtxR protein to oligonucleotides containing the 19-bp motifs was demonstrated in vitro by DNA band shift assays. At least 64 genes encoding a variety of physiological functions in iron transport and utilization, in central carbohydrate metabolism and in transcriptional regulation are controlled directly by the DtxR protein. A comparison with the bioinformatically predicted networks of C. efficiens, C. diphtheriae and C. jeikeium identified evolutionary conserved elements of the DtxR network. CONCLUSION: This work adds considerably to our currrent understanding of the transcriptional regulatory network of C. glutamicum genes that are controlled by DtxR. The DtxR protein has a major role in controlling the expression of genes involved in iron metabolism and exerts a dual regulatory function as repressor of genes participating in iron uptake and utilization and as activator of genes responsible for iron storage and DNA protection. The data suggest that the DtxR protein acts as global regulator by controlling the expression of other regulatory proteins that might take care of an iron-dependent regulation of a broader transcriptional network of C. glutamicum genes. PMID- 16469102 TI - Adaptive evolution of chloroplast genome structure inferred using a parametric bootstrap approach. AB - BACKGROUND: Genome rearrangements influence gene order and configuration of gene clusters in all genomes. Most land plant chloroplast DNAs (cpDNAs) share a highly conserved gene content and with notable exceptions, a largely co-linear gene order. Conserved gene orders may reflect a slow intrinsic rate of neutral chromosomal rearrangements, or selective constraint. It is unknown to what extent observed changes in gene order are random or adaptive. We investigate the influence of natural selection on gene order in association with increased rate of chromosomal rearrangement. We use a novel parametric bootstrap approach to test if directional selection is responsible for the clustering of functionally related genes observed in the highly rearranged chloroplast genome of the unicellular green alga Chlamydomonas reinhardtii, relative to ancestral chloroplast genomes. RESULTS: Ancestral gene orders were inferred and then subjected to simulated rearrangement events under the random breakage model with varying ratios of inversions and transpositions. We found that adjacent chloroplast genes in C. reinhardtii were located on the same strand much more frequently than in simulated genomes that were generated under a random rearrangement processes (increased sidedness; p < 0.0001). In addition, functionally related genes were found to be more clustered than those evolved under random rearrangements (p < 0.0001). We report evidence of co-transcription of neighboring genes, which may be responsible for the observed gene clusters in C. reinhardtii cpDNA. CONCLUSION: Simulations and experimental evidence suggest that both selective maintenance and directional selection for gene clusters are determinants of chloroplast gene order. PMID- 16469104 TI - Barriers and incentives to orphan care in a time of AIDS and economic crisis: a cross-sectional survey of caregivers in rural Zimbabwe. AB - BACKGROUND: Africa is in an orphan-care crisis. In Zimbabwe, where one-fourth of adults are HIV-positive and one-fifth of children are orphans, AIDS and economic decline are straining society's ability to care for orphans within their extended families. Lack of stable care is putting thousands of children at heightened risk of malnourishment, emotional underdevelopment, illiteracy, poverty, sexual exploitation, and HIV infection, endangering the future health of the society they are expected to sustain. METHODS: To explore barriers and possible incentives to orphan care, a quantitative cross-sectional survey in rural eastern Zimbabwe asked 371 adults caring for children, including 212 caring for double orphans, about their well-being, needs, resources, and perceptions and experiences of orphan care. RESULTS: Survey responses indicate that: 1) foster caregivers are disproportionately female, older, poor, and without a spouse; 2) 98% of non-foster caregivers are willing to foster orphans, many from outside their kinship network; 3) poverty is the primary barrier to fostering; 4) financial, physical, and emotional stress levels are high among current and potential fosterers; 5) financial need may be greatest in single-orphan AIDS impoverished households; and 6) struggling families lack external support. CONCLUSION: Incentives for sustainable orphan care should focus on financial assistance, starting with free schooling, and development of community mechanisms to identify and support children in need, to evaluate and strengthen families' capacity to provide orphan care, and to initiate and support placement outside the family when necessary. PMID- 16469105 TI - Chemical immobilization of adult female Weddell seals with tiletamine and zolazepam: effects of age, condition and stage of lactation. AB - BACKGROUND: Chemical immobilization of Weddell seals (Leptonychotes weddellii) has previously been, for the most part, problematic and this has been mainly attributed to the type of immobilizing agent used. In addition to individual sensitivity, physiological status may play an important role. We investigated the use of the intravenous administration of a 1:1 mixture of tiletamine and zolazepam (Telazol) to immobilize adult females at different points during a physiologically demanding 5-6 week lactation period. We also compared performance between IV and IM injection of the same mixture. RESULTS: The tiletamine:zolazepam mixture administered intravenously was an effective method for immobilization with no fatalities or pronounced apnoeas in 106 procedures; however, there was a 25 % (one animal in four) mortality rate with intramuscular administration. Induction time was slightly longer for females at the end of lactation (54.9 +/- 2.3 seconds) than at post-parturition (48.2 +/- 2.9 seconds). In addition, the number of previous captures had a positive effect on induction time. There was no evidence for effects due to age, condition (total body lipid), stage of lactation or number of captures on recovery time. CONCLUSION: We suggest that intravenous administration of tiletamine and zolazepam is an effective and safe immobilizing agent for female Weddell seals. Although individual traits could not explain variation in recovery time, we suggest careful monitoring of recovery times during longitudinal studies (> 2 captures). We show that physiological pressures do not substantially affect response to chemical immobilization with this mixture; however, consideration must be taken for differences that may exist for immobilization of adult males and juveniles. Nevertheless, we recommend a mass-specific dose of 0.50-0.65 mg/kg for future procedures with adult female Weddell seals and a starting dose of 0.50 mg/kg for other age classes and other phocid seals. PMID- 16469106 TI - Laboratory-based surveillance of current antimicrobial resistance patterns and trends among Staphylococcus aureus: 2005 status in the United States. AB - BACKGROUND: The virulence, antimicrobial resistance, and prevalence of S. aureus underscores the need for up-to-date and extensive insights regarding antimicrobial susceptibility trends. One approach to meet this need is analysis of clinical laboratory-based surveillance data. METHODS: Data from The Surveillance Network-USA (TSN), an electronic surveillance network that collects microbiology data from 300 clinical microbiology laboratories across the United States, were used as the source for analysis that included prevalence of S. aureus in clinical specimens, MRSA and multi-drug resistance phenotype rates and trends according to patient location, geographic distributions, and specimen source. RESULTS: S. aureus was the most prevalent species isolated from inpatient specimens (18.7% of all bacterial isolates) and the second most prevalent (14.7%) from outpatient specimens. In March 2005 MRSA rates were 59.2%, 55%, and 47.9% for strains from non-ICU inpatients, ICU, and outpatients, respectively. This trend was noted in all nine US Bureau of Census regions and multi-drug resistance phenotypes (resistance to > or = 3 non-beta-lactams) was common among both inpatient MRSA (59.9%) and outpatient MRSA (40.8%). Greater than 90% of multi drug resistant MRSA were susceptible to trimethoprim-sulfamethoxazole, linezolid, and vancomycin. CONCLUSION: Prevalence of MRSA among both inpatient and outpatient specimens continues to increase with multi-drug resistance as a common phenotype. Continued emergence of outpatient MRSA that exhibit multi-drug resistant phenotypes has important implications for developing and evolving outpatient treatment guidelines. PMID- 16469107 TI - The match between motivation and performance management of health sector workers in Mali. AB - Human resources for health (HRH) play a central role in improving accessibility to services and quality of care. Their motivation influences this. In Mali, operational research was conducted to identify the match between motivation and the range and use of performance management activities. OBJECTIVES: To describe the factors motivating and demotivating health workers in Mali and match the motivators with the implementation of performance management. METHODS: First an exploratory qualitative study was conducted: 28 interviews and eight group discussions were held. This was followed by a cross-sectional survey, during which 370 health workers were interviewed. The study population consisted of health workers of eight professional groups. The following issues were investigated: motivating and demotivating factors; experiences with performance management, including: job descriptions, continuous education, supervision, performance appraisal and career development. FINDINGS: The study showed that the main motivators of health workers were related to responsibility, training and recognition, next to salary. These can be influenced by performance management (job descriptions, supervisions, continuous education and performance appraisal). Performance management is not optimally implemented in Mali, as job descriptions were not present or were inappropriate; only 13% of interviewees received 4x per year supervision, and training needs were not analysed. Some 48% of the interviewees knew their performance had been appraised in the last two years; the appraisals were perceived as subjective. No other methods were in place to show recognition. The results enabled the research team to propose adaptations or improvements upon existing performance management. CONCLUSION: The results showed the importance of adapting or improving upon performance management strategies to influence staff motivation. This can be done by matching performance management activities to motivators identified by operational research. PMID- 16469108 TI - Study protocol to investigate the effect of a lifestyle intervention on body weight, psychological health status and risk factors associated with disease recurrence in women recovering from breast cancer treatment [ISRCTN08045231]. AB - BACKGROUND: Breast cancer survivors often encounter physiological and psychological problems related to their diagnosis and treatment that can influence long-term prognosis. The aim of this research is to investigate the effects of a lifestyle intervention on body weight and psychological well-being in women recovering from breast cancer treatment, and to determine the relationship between changes in these variables and biomarkers associated with disease recurrence and survival. METHODS/DESIGN: Following ethical approval, a total of 100 patients will be randomly assigned to a lifestyle intervention (incorporating dietary energy restriction in conjunction with aerobic exercise training) or normal care control group. Patients randomised to the dietary and exercise intervention will be given individualised healthy eating dietary advice and written information and attend moderate intensity aerobic exercise sessions on three to five days per week for a period of 24 weeks. The aim of this strategy is to induce a steady weight loss of up to 0.5 Kg each week. In addition, the overall quality of the diet will be examined with a view to (i) reducing the dietary intake of fat to approximately 25% of the total calories, (ii) eating at least 5 portions of fruit and vegetables a day, (iii) increasing the intake of fibre and reducing refined carbohydrates, and (iv) taking moderate amounts of alcohol. Outcome measures will include body weight and body composition, psychological health status (stress and depression), cardiorespiratory fitness and quality of life. In addition, biomarkers associated with disease recurrence, including stress hormones, estrogen status, inflammatory markers and indices of innate and adaptive immune function will be monitored. DISCUSSION: This research will provide valuable information on the effectiveness of a practical, easily implemented lifestyle intervention for evoking positive effects on body weight and psychological well-being, two important factors that can influence long-term prognosis in breast cancer survivors. However, the added value of the study is that it will also evaluate the effects of the lifestyle intervention on a range of biomarkers associated with disease recurrence and survival. Considered together, the results should improve our understanding of the potential role that lifestyle-modifiable factors could play in saving or prolonging lives. PMID- 16469109 TI - Evaluation and selection of tandem repeat loci for a Brucella MLVA typing assay. AB - BACKGROUND: The classification of Brucella into species and biovars relies on phenotypic characteristics and sometimes raises difficulties in the interpretation of the results due to an absence of standardization of the typing reagents. In addition, the resolution of this biotyping is moderate and requires the manipulation of the living agent. More efficient DNA-based methods are needed, and this work explores the suitability of multiple locus variable number tandem repeats analysis (MLVA) for both typing and species identification. RESULTS: Eighty tandem repeat loci predicted to be polymorphic by genome sequence analysis of three available Brucella genome sequences were tested for polymorphism by genotyping 21 Brucella strains (18 reference strains representing the six 'classical' species and all biovars as well as 3 marine mammal strains currently recognized as members of two new species). The MLVA data efficiently cluster the strains as expected according to their species and biovar. For practical use, a subset of 15 loci preserving this clustering was selected and applied to the typing of 236 isolates. Using this MLVA-15 assay, the clusters generated correspond to the classical biotyping scheme of Brucella spp. The 15 markers have been divided into two groups, one comprising 8 user-friendly minisatellite markers with a good species identification capability (panel 1) and another complementary group of 7 microsatellite markers with higher discriminatory power (panel 2). CONCLUSION: The MLVA-15 assay can be applied to large collections of Brucella strains with automated or manual procedures, and can be proposed as a complement, or even a substitute, of classical biotyping methods. This is facilitated by the fact that MLVA is based on non-infectious material (DNA) whereas the biotyping procedure itself requires the manipulation of the living agent. The data produced can be queried on a dedicated MLVA web service site. PMID- 16469110 TI - Bayesian detection of periodic mRNA time profiles without use of training examples. AB - BACKGROUND: Detection of periodically expressed genes from microarray data without use of known periodic and non-periodic training examples is an important problem, e.g. for identifying genes regulated by the cell-cycle in poorly characterised organisms. Commonly the investigator is only interested in genes expressed at a particular frequency that characterizes the process under study but this frequency is seldom exactly known. Previously proposed detector designs require access to labelled training examples and do not allow systematic incorporation of diffuse prior knowledge available about the period time. RESULTS: A learning-free Bayesian detector that does not rely on labelled training examples and allows incorporation of prior knowledge about the period time is introduced. It is shown to outperform two recently proposed alternative learning-free detectors on simulated data generated with models that are different from the one used for detector design. Results from applying the detector to mRNA expression time profiles from S. cerevisiae showsthat the genes detected as periodically expressed only contain a small fraction of the cell cycle genes inferred from mutant phenotype. For example, when the probability of false alarm was equal to 7%, only 12% of the cell-cycle genes were detected. The genes detected as periodically expressed were found to have a statistically significant overrepresentation of known cell-cycle regulated sequence motifs. One known sequence motif and 18 putative motifs, previously not associated with periodic expression, were also over represented. CONCLUSION: In comparison with recently proposed alternative learning-free detectors for periodic gene expression, Bayesian inference allows systematic incorporation of diffuse a priori knowledge about, e.g. the period time. This results in relative performance improvements due to increased robustness against errors in the underlying assumptions. Results from applying the detector to mRNA expression time profiles from S. cerevisiae include several new findings that deserve further experimental studies. PMID- 16469111 TI - CBFbeta is a facultative Runx partner in the sea urchin embryo. AB - BACKGROUND: Runx proteins are developmentally important metazoan transcription factors that form a heterodimeric complex with the non-homologous protein Core Binding Factor beta (CBFbeta). CBFbeta allosterically enhances Runx DNA binding but does not bind DNA itself. We report the initial characterization of SpCBFbeta, the heterodimeric partner of SpRunt-1 from the sea urchin Stronylocentrotus purpuratus. RESULTS: SpCBFbeta is remarkably similar to its mammalian homologues, and like them it enhances the DNA binding of the Runt domain. SpCBFbeta is entirely of zygotic provenance and its expression is similar that of SpRunt-1, accumulating globally at late blastula stage then later localizing to endoderm and oral ectoderm. Unlike SpRunt-1, however, SpCBFbeta is enriched in the endodermal mid- and hindgut of the pluteus larva, and is not highly expressed in the foregut and ciliated band. We showed previously that morpholino antisense-mediated knockdown of SpRunt-1 leads to differentiation defects, as well as to extensive post-blastula stage apoptosis caused by under expression of the Runx target gene SpPKC1. In contrast, we show here that knockdown of SpCBFbeta does not negatively impact cell survival or SpPKC1 expression, although it does lead to differentiation defects similar to those associated with SpRunt-1 deficiency. Moreover, SpRunt-1 containing a single amino acid substitution that abolishes its ability to interact with SpCBFbeta retains the ability to rescue cell survival in SpRunt-1 morphant embryos. Chromatin immunoprecipitation shows that while the CyIIIa promoter engages both proteins, the SpPKC1 promoter only engages SpRunt-1. CONCLUSION: SpCBFbeta is a facultative Runx partner that appears to be required specifically for cell differentiation. PMID- 16469112 TI - First-dose analgesic effect of the cyclo-oxygenase-2 selective inhibitor lumiracoxib in osteoarthritis of the knee: a randomized, double-blind, placebo controlled comparison with celecoxib [NCT00267215]. AB - Cyclo-oxygenase-2 selective inhibitors are frequently used to manage osteoarthritis. We compared the analgesic efficacy of the novel cyclo-oxygenase-2 selective inhibitor lumiracoxib (Prexige) versus placebo and celecoxib in patients with knee osteoarthritis. This seven day, double-blind, placebo and active comparator controlled, parallel group study included 364 patients aged > or = 50 years with moderate-to-severe symptomatic knee osteoarthritis. Patients received lumiracoxib 400 mg/day (four times the recommended chronic dose in osteoarthritis; n = 144), placebo (n = 75), or celecoxib 200 mg twice daily (n = 145). The primary variable was actual pain intensity difference (100 mm visual analogue scale) between baseline and the mean of three hour and five hour assessments after the first dose. Actual pain intensity difference, average and worst pain, pain relief and functional status (Western Ontario and McMaster Universities Osteoarthritis Index [WOMAC]) were measured over seven days. Patients also completed a global evaluation of treatment effect at study end or premature discontinuation. For the primary variable, the superiority of lumiracoxib versus placebo, the noninferiority of lumiracoxib versus celecoxib, and the superiority of lumiracoxib versus celecoxib were assessed by closed test procedure adjusting for multiplicity, thereby maintaining the overall 5% significance level. In addition, celecoxib was assessed versus placebo in a predefined exploratory manner to assess trial sensitivity. Lumiracoxib provided better analgesia than placebo 3-5 hours after the first dose (P = 0.004) through to study end. The estimated difference between lumiracoxib and celecoxib 3-5 hours after the first dose was not significant (P = 0.185). Celecoxib was not significantly different from placebo in this analysis (P = 0.069). At study end 13.9% of lumiracoxib-treated patients reported complete pain relief versus 5.5% and 5.3% of celecoxib and placebo recipients, respectively. WOMAC total and subscales improved for both active treatments versus placebo except for difficulty in performing daily activities, for which celecoxib just failed to achieve significance (P = 0.056). In the patient's global evaluation of treatment effect, 58.1% of patients receiving lumiracoxib rated treatment as 'excellent' or 'good', versus 48.6% of celecoxib and 25.3% of placebo patients. Lumiracoxib was well tolerated. The overall incidence of adverse events was similar across treatment groups. PMID- 16469113 TI - Detailed analysis of the variability of peptidylarginine deiminase type 4 in German patients with rheumatoid arthritis: a case-control study. AB - Peptidylarginine deiminase type 4 (PADI4) genotypes were shown to influence susceptibility to rheumatoid arthritis (RA) in the Japanese population. Such an association could not previously be confirmed in different European populations. In the present study, we analysed exons 2-4 of PADI4 in 102 German RA patients and 102 healthy individuals to study the influence of PADI4 variability on RA susceptibility by means of haplotype-specific DNA sequencing. Analyses of the influence of PADI4 and HLA-DRB1 genotypes on disease activity and on levels of anti-cyclic citrullinated peptide antibodies were performed. Comparing the frequencies of PADI4 haplotype 4 (padi4_89*G, padi4_90*T, padi4_92*G, padi4_94*T, padi4_104*C, padi4_95*G, padi4_96*T) (patients, 14.7%; controls, 7.8%; odds ratio = 2.0, 95% confidence interval = 1.1-3.8) and carriers of this haplotype (patients, 27.5%; controls, 13.7%; odds ratio = 2.4, 95% confidence interval = 1.2-4.8), a significant positive association of PADI4 haplotype 4 with RA could be demonstrated. Other PADI4 haplotypes did not differ significantly between patients and controls. Regarding the individual PADI4 variants, padi4_89 (A-->G), padi4_90 (C-->T), and padi4_94 (C-->T) were significantly associated with RA (patients, 49.5%; controls, 38.7%; odds ratio = 1.6, 95% confidence interval = 1.1-2.3). Considering novel PADI4 variants located in or near to exons 2, 3, and 4, no quantitative or qualitative differences between RA patients (8.8%) and healthy controls (10.8%) could be demonstrated. While the PADI4 genotype did not influence disease activity and the anti-cyclic citrullinated peptide antibody level, the presence of the HLA-DRB1 shared epitope was significantly associated with higher anti-cyclic citrullinated peptide antibody levels (P = 0.033). The results of this small case-control study support the hypothesis that variability of the PADI4 gene may influence susceptibility to RA in the German population. Quantitative or qualitative differences in previously undefined PADI4 variants between patients and controls could not be demonstrated. PMID- 16469114 TI - The induction of CCN2 by TGFbeta1 involves Ets-1. AB - CCN2 is encoded by an immediate-early gene induced in mesenchymal cells during the formation of blood vessels, bone and connective tissue. It plays key roles in cell adhesion and migration, as well as matrix remodeling. CCN2 is overexpressed in fibrosis, arthritis and cancer; thus, an understanding of how to control CCN2 expression is likely to have importance in developing therapies to combat these pathologies. Previously, we found that the promoter sequence GAGGAATG is important for Ccn2 gene regulation in NIH 3T3 fibroblasts. In this report, we show that this sequence mediates activation of the CCN2 promoter by the ETS family of transcription factors. Endogenous Ets-1 binds this element of the CCN2 promoter, and dominant negative Ets-1 and specific Ets-1 small interfering RNA block induction of CCN2 expression by TGFbeta. In the absence of added TGFbeta1, Ets-1, but not the related fli-1, synergizes with Smad 3 to activate the CCN2 promoter. Whereas the ability of transfected Ets-1 to activate the CCN2 promoter is dependent on protein kinase C (PKC), Ets-1 in the presence of co-transfected Smad3 does not require PKC, suggesting that the presence of Smad3 bypasses the requirement of Ets-1 for PKC to activate target promoter activity. Our results are consistent with the notion that Smad3 and Ets-1 cooperate in the induction of the CCN2 promoter by TGFbeta1. Antagonizing Ets-1 might be of benefit in attenuating CCN2 expression in fibrosis, arthritis and cancer, and may be useful in modulating the outcome of these disorders. PMID- 16469115 TI - CD95-induced osteoarthritic chondrocyte apoptosis and necrosis: dependency on p38 mitogen-activated protein kinase. AB - One of the hallmarks of osteoarthritic cartilage is the loss of chondrocyte cellularity due to cell death. However, considerable controversy has recently arisen surrounding the extent of apoptotic cell death involved in development of osteoarthritis (OA). To shed light on this issue, we characterized cell death in primary OA chondrocytes mediated by the CD95 (Fas) pathway. Treatment of chondrocytes with anti-CD95 not only increased the rate of cell death but also increased the production of CD95 ligand by chondrocytes. This reveals a novel autocrine regulatory loop whereby activated chondrocytes may amplify CD95 signals by inducing synthesis of CD95 ligand. Multiple morphologic detection analyses indicated that apoptosis accounted for only a portion of chondrocyte death, whereas the other chondrocytes died by necrosis. Both chondrocyte apoptosis and necrosis depended on the activity of p38 mitogen-activated protein kinase (MAPK) within chondrocytes. Treatment of chondrocytes with the p38 MAPK inhibitor SB203580 abolished anti-CD95 induced cell death by inhibiting the activities of activating transcription factor-2 and caspase-3. In addition, inhibition of p38 MAPK activity in chondrocytes stimulated chondrocyte proliferation, as indicated by 5-bromo-2-deoxyuridine (BrdU) index. Thus, p38 MAPK is a potential therapeutic target, inhibition of which may maintain the cellularity of articular chondrocytes by inhibiting cell death and its amplification signal and by increasing cell proliferation. PMID- 16469116 TI - Association between microscopic brain damage as indicated by magnetization transfer imaging and anticardiolipin antibodies in neuropsychiatric lupus. AB - The pathogenetic role of anticardiolipin antibodies (aCLs) in patients with neuropsychiatric systemic lupus erythematosus (NPSLE) without cerebral infarcts remains elusive. Magnetization transfer imaging (MTI) has proved to be a sensitive tool for detecting diffuse microscopic brain damage in NPSLE patients. In this study we examined the correlation between grey and white matter magnetization transfer ratio (MTR) parameters and the presence of IgM and IgG aCLs and lupus anticoagulant in 18 patients with systemic lupus erythematosus and a history of NPSLE but without cerebral infarcts on conventional magnetic resonance imaging. Lower grey matter mean MTR (P < 0.05), white matter mean MTR (P < 0.05), white matter peak location (P < 0.05) and grey matter peak location (trend toward statistical significance) were observed in IgM aCL-positive patients than in IgM aCL-negative patients. No significant differences were found in MTR histogram parameters with respect to IgG aCL and lupus anticoagulant status, nor with respect to anti-dsDNA or anti-ENA (extractable nuclear antigen) status. This is the first report of an association between the presence of aCLs and cerebral damage in grey and white matter in NPSLE. Our findings suggest that aCLs are associated with diffuse brain involvement in NPSLE patients. PMID- 16469117 TI - Human autoantibodies against the 54 kDa protein of the signal recognition particle block function at multiple stages. AB - The 54 kDa subunit of the signal recognition particle (SRP54) binds to the signal sequences of nascent secretory and membrane proteins and it contributes to the targeting of these precursors to the membrane of the endoplasmic reticulum (ER). At the ER membrane, the binding of the signal recognition particle (SRP) to its receptor triggers the release of SRP54 from its bound signal sequence and the nascent polypeptide is transferred to the Sec61 translocon for insertion into, or translocation across, the ER membrane. In the current article, we have characterized the specificity of anti-SRP54 autoantibodies, which are highly characteristic of polymyositis patients, and investigated the effect of these autoantibodies on the SRP function in vitro. We found that the anti-SRP54 autoantibodies had a pronounced and specific inhibitory effect upon the translocation of the secretory protein preprolactin when analysed using a cell free system. Our mapping studies showed that the anti-SRP54 autoantibodies bind to the amino-terminal SRP54 N-domain and to the central SRP54 G-domain, but do not bind to the carboxy-terminal M-domain that is known to bind ER signal sequences. Nevertheless, anti-SRP54 autoantibodies interfere with signal-sequence binding to SRP54, most probably by steric hindrance. When the effect of anti-SRP autoantibodies on protein targeting the ER membrane was further investigated, we found that the autoantibodies prevent the SRP receptor-mediated release of ER signal sequences from the SRP54 subunit. This observation supports a model where the binding of the homologous GTPase domains of SRP54 and the alpha-subunit of the SRP receptor to each other regulates the release of ER signal sequences from the SRP54 M-domain. PMID- 16469118 TI - Serial determination of cyclic citrullinated peptide autoantibodies predicted five-year radiological outcomes in a prospective cohort of patients with early rheumatoid arthritis. AB - The objective of this study was to evaluate the potential of serially determined anti-cyclic citrullinated peptide (CCP) antibodies for predicting structural joint damage in patients with early rheumatoid arthritis (RA), compared to a single baseline determination. Ninety-nine RA patients with disease durations of less than one year and no history of disease-modifying antirheumatic drug therapy were followed prospectively for at least five years. Anti-CCP2 concentrations were measured using a second-generation ELISA. Sharp scores as modified by van der Heijde were determined on hand and foot radiographs. Anti-CCP2 antibodies were detected in 55.5% of patients at baseline and 63.6% at any time during the first three years. Presence of anti-CCP2 at any time during the first three years was associated with radiographic damage at baseline (odds ratio (OR), 3.66; 95% confidence interval (95% CI) 0.99-13.54) and with five year progression of the total Sharp score (OR, 3.17; 95% CI, 1.3-7.7), erosion score (OR, 5.3; 95% CI, 1.4-19.2) and joint space narrowing score (OR, 2.8; 95% CI, 1.15-6.8). The presence of anti-CCP2 or IgM RF at baseline did not predict these outcomes. Patients with negative anti-CCP2 tests throughout follow-up had less radiographic progression than patients with increasing anti-CCP2 concentrations; they did not differ from patients with decreasing anti-CCP2 antibody levels. HLADRB1* typing showed that progression of the mean modified Sharp score was not correlated with the presence of the shared epitope alleles. In conclusion, serially determined anti-CCP2 antibodies during the first three years of follow-up performs better than baseline determination for predicting radiographic progression in patients with early RA. PMID- 16469120 TI - Dendritic cell defects in patients with cancer: mechanisms and significance. AB - Dendritic cells (DCs) are a complex network of antigen-presenting cells that have an essential role in the modulation of primary immunity. There has been increasing evidence that DCs isolated from patients with malignancy demonstrate functional deficiencies that inhibit the capacity to mount an effective anti tumor response. In this issue of Breast Cancer Research, Pinzon-Charry and colleagues investigate one of the possible mechanisms by which tumors induce DC dysfunction to evade host immune surveillance. They demonstrate that DCs isolated from the circulation of patients with early-stage breast cancer exhibit increased rates of spontaneous apoptosis. In vitro studies suggest that a soluble factor secreted by breast cancer cells is responsible for this phenomenon. In contrast, ex vivo conditioning of DCs with CD-40 ligand and IL-12 was protective against tumor-induced apoptosis. PMID- 16469119 TI - Proteoglycan 4 downregulation in a sheep meniscectomy model of early osteoarthritis. AB - Osteoarthritis is a disease of multifactorial aetiology characterised by progressive breakdown of articular cartilage. In the early stages of the disease, changes become apparent in the superficial zone of articular cartilage, including fibrillation and fissuring. Normally, a monolayer of lubricating molecules is adsorbed on the surface of cartilage and contributes to the minimal friction and wear properties of synovial joints. Proteoglycan 4 is the lubricating glycoprotein believed to be primarily responsible for this boundary lubrication. Here we have used an established ovine meniscectomy model of osteoarthritis, in which typical degenerative changes are observed in the operated knee joints at three months after surgery, to evaluate alterations in proteoglycan 4 expression and localisation in the early phases of the disease. In normal control joints, proteoglycan 4 was immunolocalised in the superficial zone of cartilage, particularly in those regions of the knee joint covered by a meniscus. After the onset of early osteoarthritis, we demonstrated a loss of cellular proteoglycan 4 immunostaining in degenerative articular cartilage, accompanied by a significant (p < 0.01) decrease in corresponding mRNA levels. Early loss of proteoglycan 4 from the cartilage surface in association with a decrease in its expression by superficial-zone chondrocytes might have a role in the pathogenesis of osteoarthritis. PMID- 16469121 TI - Recent translational research: oncogene discovery by insertional mutagenesis gets a new boost. AB - Knowledge of the genes and genetic pathways involved in onco-genesis is essential if we are to identify novel targets for cancer therapy. Insertional mutagenesis in mouse models is among the most efficient tools to detect novel cancer genes. Retrovirus-mediated insertional mutagenesis received a tremendous boost by the availability of the mouse genome sequence and new PCR methods. Application of such advances were limited to lympho-magenesis but are now also being applied to mammary tumourigenesis. Novel transposons that allow insertional mutagenesis studies to be conducted in tumors of any mouse tissue may give cancer gene discovery a further boost. PMID- 16469122 TI - Do statins have a role in preventing or treating sepsis? AB - Statins have a variety of properties that are independent of their lipid lowering ability. These anti-inflammatory, antioxidant, immunomodulatory, and antiapoptotic features have been collectively referred to as pleiotropic effects. Severe sepsis is an intense infection-induced inflammatory syndrome that ultimately results in organ dysfunction. Because so many cascades are triggered during sepsis, merely blocking a single component may be insufficient to arrest the inflammatory process. A growing body of evidence suggests that statins may indeed have a protective effect against severe sepsis and reduce the rate of infection-related mortality. This novel primary prevention concept may have far reaching implications for the future management of serious infections. Moreover, it was recently shown that statins potentially improve outcome after the onset of sepsis. The stage is now set for randomized clinical trials that will determine the precise role, if any, that statins may have in preventing and treating sepsis. PMID- 16469123 TI - Choosing the right combination therapy in severe community-acquired pneumonia. AB - Recent studies have suggested that combination antibiotic therapy is preferable to monotherapy for severe community-acquired pneumonia (CAP). In this issue Mortensen and colleagues present retrospective data suggesting that combination therapy with a cephalosporin and a fluoroquinolone is inferior to combination therapy with a cephalosporin and a macrolide. Several mechanisms exist by which quinolones could be inferior to macrolides in combination therapy, so if these findings are confirmed by other groups they have significant implications for physicians treating patients with severe CAP. PMID- 16469124 TI - Towards a feasible algorithm for tight glycaemic control in critically ill patients: a systematic review of the literature. AB - INTRODUCTION: Tight glycaemic control is an important issue in the management of intensive care unit (ICU) patients. The glycaemic goals described by Van Den Berghe and colleagues in their landmark study of intensive insulin therapy appear difficult to achieve in a real life ICU setting. Most clinicians and nurses are concerned about a potentially increased frequency of severe hypoglycaemic episodes with more stringent glycaemic control. One of the steps we took before we implemented a glucose regulation protocol was to review published trials employing insulin/glucose algorithms in critically ill patients. METHODS: We conducted a search of the PubMed, Embase and Cochrane databases using the following terms: 'glucose', 'insulin', 'protocol', 'algorithm', 'nomogram', 'scheme', 'critically ill' and 'intensive care'. Our search was limited to clinical trials conducted in humans. The aim of the papers selected was required to be glycaemic control in critically ill patients; the blood glucose target was required to be 10 mmol/l or under (or use of a protocol that resulted in a mean blood glucose = 10 mmol/l). The studies were categorized according to patient type, desired range of blood glucose values, method of insulin administration, frequency of blood glucose control, time taken to achieve the desired range for glucose, proportion of patients with glucose in the desired range, mean blood glucose and frequency of hypoglycaemic episodes. RESULTS: A total of twenty-four reports satisfied our inclusion criteria. Most recent studies (nine) were conducted in an ICU; nine others were conducted in a perioperative setting and six were conducted in patients with acute myocardial infarction or stroke. Studies conducted before 2001 did not include normoglycaemia among their aims, which changed after publication of the study by Van Den Berghe and coworkers in 2001; glycaemic goals became tighter, with a target range between 4 and 8 mmol/l in most studies. CONCLUSION: Studies using a dynamic scale protocol combining a tight glucose target and the last two blood glucose values to determine the insulin infusion rate yielded the best results in terms of glycaemic control and reported low frequencies of hypoglycaemic episodes. PMID- 16469126 TI - Impact of computerized physician order entry on medication prescription errors in the intensive care unit: a controlled cross-sectional trial. AB - INTRODUCTION: Medication errors in the intensive care unit (ICU) are frequent and lead to attributable patient morbidity and mortality, increased length of ICU stay and substantial extra costs. We investigated if the introduction of a computerized ICU system (Centricity Critical Care Clinisoft, GE Healthcare) reduced the incidence and severity of medication prescription errors (MPEs). METHODS: A prospective trial was conducted in a paper-based unit (PB-U) versus a computerized unit (C-U) in a 22-bed ICU of a tertiary university hospital. Every medication order and medication prescription error was validated by a clinical pharmacist. The registration of different classes of MPE was done according to the National Coordinating Council for Medication Error Reporting and Prevention guidelines. An independent panel evaluated the severity of MPEs. We identified three groups: minor MPEs (no potential to cause harm); intercepted MPEs (potential to cause harm but intercepted on time); and serious MPEs (non intercepted potential adverse drug events (ADE) or ADEs, being MPEs with potential to cause, or actually causing, patient harm). RESULTS: The C-U and the PB-U each contained 80 patient-days, and a total of 2,510 medication prescriptions were evaluated. The clinical pharmacist identified 375 MPEs. The incidence of MPEs was significantly lower in the C-U compared with the PB-U (44/1286 (3.4%) versus 331/1224 (27.0%); P < 0.001). There were significantly less minor MPEs in the C-U than in the PB-U (9 versus 225; P < 0.001). Intercepted MPEs were also lower in the C-U (12 versus 46; P < 0.001), as well as the non-intercepted potential ADEs (21 versus 48; P < 0.001). There was also a reduction of ADEs (2 in the C-U versus 12 in the PB-U; P < 0.01). No fatal errors occurred. The most frequent drug classes involved were cardiovascular medication and antibiotics in both groups. Patients with renal failure experienced less dosing errors in the C-U versus the PB-U (12 versus 35 serious MPEs; P < 0.001). CONCLUSION: The ICU computerization, including the medication order entry, resulted in a significant decrease in the occurrence and severity of medication errors in the ICU. PMID- 16469125 TI - Evaluation of rapid screening and pre-emptive contact isolation for detecting and controlling methicillin-resistant Staphylococcus aureus in critical care: an interventional cohort study. AB - INTRODUCTION: Rapid diagnostic tests may allow early identification of previously unknown methicillin-resistant Staphylococcus aureus (MRSA) carriers at intensive care unit (ICU) admission. The aim of this study was twofold: first, to assess whether a new molecular MRSA screening test can substantially decrease the time between ICU admission and identification of MRSA carriers; and, second, to examine the combined effect of rapid testing and pre-emptive contact isolation on MRSA infections. METHOD: Since November 2003, patients admitted for longer than 24 hours to two adult ICUs were screened systematically on admission using quick, multiplex immunocapture-coupled PCR (qMRSA). Median time intervals from admission to notification of test results were calculated for a five-month intervention phase (November 2003-March 2004) and compared with a historical control period (April 2003-October 2003) by nonparametric tests. ICU-acquired MRSA infection rates were determined for an extended surveillance period (January 2003 through August 2005) and analyzed by Poisson regression methods. RESULTS: During the intervention phase, 97% (450/462) of patients admitted to the surgical ICU and 80% (470/591) of patients admitted to the medical ICU were screened. On-admission screening identified the prevalence of MRSA to be 6.7% (71/1053). Without admission screening, 55 previously unknown MRSA carriers would have been missed in both ICUs. Median time from ICU admission to notification of test results decreased from 87 to 21 hours in the surgical ICU (P < 0.001) and from 106 to 23 hours in the medical ICU (P < 0.001). In the surgical ICU, 1,227 pre-emptive isolation days for 245 MRSA-negative patients were saved by using the qMRSA test. After adjusting for colonization pressure, the systematic on-admission screening and pre-emptive isolation policy was associated with a reduction in medical ICU acquired MRSA infections (relative risk 0.3, 95% confidence interval 0.1-0.7) but had no effect in the surgical ICU (relative risk 1.0, 95% confidence interval 0.6 1.7). CONCLUSION: The qMRSA test decreased median time to notification from four days to one day and helped to identify previously unknown MRSA carriers rapidly. A strategy linking the rapid screening test to pre-emptive isolation and cohorting of MRSA patients substantially reduced MRSA cross-infections in the medical but not in the surgical ICU. PMID- 16469127 TI - Rescue treatment with terlipressin in children with refractory septic shock: a clinical study. AB - INTRODUCTION: Refractory septic shock has dismal prognosis despite aggressive therapy. The purpose of the present study is to report the effects of terlipressin (TP) as a rescue treatment in children with catecholamine refractory hypotensive septic shock. METHODS: We prospectively registered the children with severe septic shock and hypotension resistant to standard intensive care, including a high dose of catecholamines, who received compassionate therapy with TP in nine pediatric intensive care units in Spain, over a 12-month period. The TP dose was 0.02 mg/kg every four hours. RESULTS: Sixteen children (age range, 1 month-13 years) were included. The cause of sepsis was meningococcal in eight cases, Staphylococcus aureus in two cases, and unknown in six cases. At inclusion the median (range) Pediatric Logistic Organ Dysfunction score was 23.5 (12-52) and the median (range) Pediatric Risk of Mortality score was 24.5 (16-43). All children had been treated with a combination of at least two catecholamines at high dose rates. TP treatment induced a rapid and sustained improvement in the mean arterial blood pressure that allowed reduction of the catecholamine infusion rate after one hour in 14 out of 16 patients. The mean (range) arterial blood pressure 30 minutes after TP administration increased from 50.5 (37-93) to 77 (42 100) mmHg (P < 0.05). The noradrenaline infusion rate 24 hours after TP treatment decreased from 2 (1-4) to 1 (0-2.5) microg/kg/min (P < 0.05). Seven patients survived to the sepsis episode. The causes of death were refractory shock in three cases, withdrawal of therapy in two cases, refractory arrhythmia in three cases, and multiorgan failure in one case. Four of the survivors had sequelae: major amputations (lower limbs and hands) in one case, minor amputations (finger) in two cases, and minor neurological deficit in one case. CONCLUSION: TP is an effective vasopressor agent that could be an alternative or complementary therapy in children with refractory vasodilatory septic shock. The addition of TP to high doses of catecholamines, however, can induce excessive vasoconstriction. Additional studies are needed to define the safety profile and the clinical effectiveness of TP in children with septic shock. PMID- 16469128 TI - Molecular adsorbent recirculating system and hemostasis in patients at high risk of bleeding: an observational study. AB - INTRODUCTION: Liver failure is associated with reduced synthesis of clotting factors, consumptive coagulopathy, and platelet dysfunction. The aim of the study was to evaluate the effects of liver support using a molecular adsorbent recirculating system (MARS) on the coagulation system in patients at high risk of bleeding. METHODS: We studied 61 MARS treatments in 33 patients with acute liver failure (n = 15), acute-on-chronic liver failure (n = 8), sepsis (n = 5), liver graft dysfunction (n = 3), and cholestasis (n = 2). Standard coagulation tests, standard thromboelastography (TEG), and heparinase-modified and abciximab-fab modified TEG were performed immediately before and 30 minutes after commencement of MARS, and after the end of MARS treatment. Prostaglandin I2 was administered extracorporeally to all patients; 17 patients additionally received unfractioned heparin. RESULTS: Three moderate bleeding complications in three patients, requiring three to four units of packed red blood cells, were observed. All were sufficiently managed without interrupting MARS treatment. Although there was a significant decrease in platelet counts (median, 9 G/l; range, -40 to 145 G/l) and fibrinogen concentration (median, 15 mg/dl; range, -119 to 185 mg/dl) with a consecutive increase in thrombin time, the platelet function, as assessed by abciximab-fab-modified TEG, remained stable. MARS did not enhance fibrinolysis. CONCLUSION: MARS treatment appears to be well tolerated during marked coagulopathy due to liver failure. Although MARS leads to a further decrease in platelet count and fibrinogen concentration, platelet function, measured as the contribution of the platelets to the clot firmness in TEG, remains stable. According to TEG-based results, MARS does not enhance fibrinolysis. PMID- 16469130 TI - Imposed work of breathing during high-frequency oscillatory ventilation: a bench study. AB - INTRODUCTION: The ventilator and the endotracheal tube impose additional workload in mechanically ventilated patients breathing spontaneously. The total work of breathing (WOB) includes elastic and resistive work. In a bench test we assessed the imposed WOB using 3100 A/3100 B SensorMedics high-frequency oscillatory ventilators. METHODS: A computer-controlled piston-driven test lung was used to simulate a spontaneously breathing patient. The test lung was connected to a high frequency oscillatory ventilation (HFOV) ventilator by an endotracheal tube. The inspiratory and expiratory airway flows and pressures at various places were sampled. The spontaneous breath rate and volume, tube size and ventilator settings were simulated as representative of the newborn to adult range. The fresh gas flow rate was set at a low and a high level. The imposed WOB was calculated using the Campbell diagram. RESULTS: In the simulations for newborns (assumed body weight 3.5 kg) and infants (assumed body weight 10 kg) the imposed WOB (mean +/- standard deviation) was 0.22 +/- 0.07 and 0.87 +/- 0.25 J/l, respectively. Comparison of the imposed WOB in low and high fresh gas flow rate measurements yielded values of 1.63 +/- 0.32 and 0.96 +/- 0.24 J/l (P = 0.01) in small children (assumed body weight 25 kg), of 1.81 +/- 0.30 and 1.10 +/- 0.27 J/l (P < 0.001) in large children (assumed body weight 40 kg), and of 1.95 +/- 0.31 and 1.12 +/- 0.34 J/l (P < 0.01) in adults (assumed body weight 70 kg). High peak inspiratory flow and low fresh gas flow rate significantly increased the imposed WOB. Mean airway pressure in the breathing circuit decreased dramatically during spontaneous breathing, most markedly at the low fresh gas flow rate. This led to ventilator shut-off when the inspiratory flow exceeded the fresh gas flow. CONCLUSION: Spontaneous breathing during HFOV resulted in considerable imposed WOB in pediatric and adult simulations, explaining the discomfort seen in those patients breathing spontaneously during HFOV. The level of imposed WOB was lower in the newborn and infant simulations, explaining why these patients tolerate spontaneous breathing during HFOV well. A high fresh gas flow rate reduced the imposed WOB. These findings suggest the need for a demand flow system based on patient need allowing spontaneous breathing during HFOV. PMID- 16469131 TI - Do fluoroquinolones actually increase mortality in community-acquired pneumonia? PMID- 16469132 TI - Cardiac troponin level is not an independent predictor of mortality in septic patients requiring medical intensive care unit admission. PMID- 16469134 TI - Recently published papers: pulmonary care, pandemics, and eugenics in surviving sepsis? AB - Respiratory failure is one of the leading admission diagnoses on the critical care unit, and the journals have reflected this over the past few months. An understanding of the aetiology of pulmonary sepsis is important but your choice of ventilator gas humidification system is not. There are prophecies of more pandemics, but panic is futile because survival is all down to your genes. PMID- 16469133 TI - Reactive oxygen species: toxic molecules or spark of life? AB - Increases in reactive oxygen species (ROS) and tissue evidence of oxidative injury are common in patients with inflammatory processes or tissue injury. This has led to many clinical attempts to scavenge ROS and reduce oxidative injury. However, we live in an oxygen rich environment and ROS and their chemical reactions are part of the basic chemical processes of normal metabolism. Accordingly, organisms have evolved sophisticated mechanisms to control these reactive molecules. Recently, it has become increasingly evident that ROS also play a role in the regulation of many intracellular signaling pathways that are important for normal cell growth and inflammatory responses that are essential for host defense. Thus, simply trying to scavenge ROS is likely not possible and potentially harmful. The 'normal' level of ROS will also likely vary in different tissues and even in different parts of cells. In this paper, the terminology and basic chemistry of reactive species are reviewed. Examples and mechanisms of tissue injury by ROS as well as their positive role as signaling molecules are discussed. Hopefully, a better understanding of the nature of ROS will lead to better planned therapeutic attempts to manipulate the concentrations of these important molecules. We need to regulate ROS, not eradicate them. PMID- 16469135 TI - Does nutrition have a role in peripheral vascular disease? AB - Peripheral vascular disease (PVD) is a manifestation of systemic atherosclerosis in the lower limbs, and PVD patients have a 3- to 5-fold increased risk of cardiovascular mortality compared with age-matched controls. Nevertheless, recent reports show how PVD patients are undertreated with regard to CVD risk-factor reduction and the use of lipid-lowering or antiplatelet drugs. There is appreciable evidence that demonstrates the beneficial effects of certain nutrients and dietary habits in the prevention of CVD, but there has been little attention paid to the role of nutrients in PVD. The purpose of the present review is to provide an overview of our understanding of how foods could possibly benefit PVD. In the last few decades, several nutrients have arisen as potentially health-promoting in PVD. While nutritional interventions in PVD show positive clinical effects for fish oil, carnitine or vitamin E, others such as olive oil or vitamin C seem to interact only at a biochemical level by decreasing risk factors. Moreover, only epidemiological associations exist for the potential role of fibre, folates or vitamin B6 in this disease. In all cases, the limited data available provide no clear-cut evidence in favour of the clinical benefit of nutritional interventions aimed at reducing risk factors and ameliorating symptoms in PVD patients. No practical recommendations can be given at this stage, and further studies are clearly needed. PMID- 16469136 TI - Cold-pressed flaxseed oil reverses age-associated depression in a primary cell mediated adaptive immune response in the mouse. AB - The objective of this investigation was to determine the influence of flaxseed oil on responses representative of primary humoral and cell-mediated adaptive immune competence in immunosenescent mice. Male and female C57BL/6J mice, 85 weeks old, were randomized between two complete purified diets differing only in oil source (cold-pressed safflower or flaxseed). After 8 weeks, humoral competence was assessed in six mice per group as the serum haemagglutinin titre to sheep red blood cells (SRBC) and cell-mediated competence was assessed, in an additional six mice per group, as the delayed hypersensitivity response to SRBC. A zero-time control group (88 weeks old) and a young adult positive control group (12 weeks old) were each tested similarly (six per immune response), revealing age-related depression in both antibody and cell-mediated competence at 88 weeks of age. After the 8-week experimental period, the antibody response of the two test groups of geriatric mice remained below the young adult level (P=0.04) and the cell-mediated response of the safflower oil group also continued to exhibit age-related depression (20 % of young adult level, P=0.0002). By contrast, the anti-SRBC delayed hypersensitivity response of the flaxseed group no longer differed from the response of the young adults but exceeded that of the safflower and zero-time control senescent groups (P=0.0002). Depression in primary cell mediated competence, the most outstanding aspect of immunosenescence, can be addressed by means of a dietary source of 18 : 3n-3 without longer-chain PUFA. PMID- 16469137 TI - Maintenance threonine requirement and efficiency of its use for accretion of whole-body threonine and protein in Atlantic salmon (Salmo salar L.) fry. AB - Eighteen groups of seventy Atlantic salmon (Salmo salar L.) fry (initial mean body weight 0.8 (sd 0.01) g) were fed on semi-purified diets containing graded levels of l-threonine (Thr) in 15 litres aquaria at a temperature of 14.5+/-1 degrees C. Doses of Thr represented 1, 31, 41, 51, 62, 72, 83 and 93 % of its ideal level for optimum protein deposition. Indispensable amino acids other than Thr were included in the same proportion (on a g/16 g N basis) as in the Atlantic salmon fry whole-body carcass. Following 36 d of feeding and a 36 h fast, fry were killed for whole-body protein and amino acid analysis. Weight gain (r2 0.98), protein accretion (r2 0.97), and Thr accretion (r2 0.97) were linear (P<0.01) functions of Thr intake. Slope of the Thr accretion regression line showed that the efficiency of Thr utilisation above maintenance was 76 %. At zero Thr intake, fry lost 5.4 mg Thr/kg body weight0.75 per d. The Thr maintenance requirement was 7.2 mg/kg body weight0.75 per d and the Thr requirement for growth was 66 mg for 1 g protein deposition. Increasing doses of Thr resulted in increased (P<0.05) concentrations of histidine and lysine, and decreased concentrations of isoleucine in whole-body protein. The maintenance need for Thr represented 13.4 % of the total need for Thr. The data suggest that efficiency of Thr utilisation above maintenance is constant at all levels of Thr intake between 1 and 93 % of the level required for optimum protein deposition. PMID- 16469138 TI - Incorporating turnover in estimates of protein retention efficiency for different body tissues. AB - Formulated in terms of protein synthesis (PS) and protein retention (PR), a definition of turnover-related protein retention efficiency (kP) allows the expression kP=[1+(PS/PR)/6](-1), 6 representing the ratio of the energy equivalent of protein to the cost of synthesis. By combining plausible hormonal and cellular control mechanisms of protein (P) growth, it is possible to derive (PS/PR)=[Q{(P/alpha)(-(4/9)Y)-1}](-1)+1, allowing the calculation of kP by substitution. The symbol alpha represents the limit value of protein growth, while the term 4/9 derives from the power in the relationship between the concentration of growth factor-related activator in the nucleus and cell volume (cv). Y is the power in the relationship between cv and total tissue protein, and Q represents the proportion of growth factor-activated nuclei in a tissue. The proportion Q can be estimated from simple functions of intake rates or blood growth factor concentrations. Estimates of Y are derived from histological considerations or calculated from experimental observations; Y=1 for multinucleated skeletal muscle fibres and Y=1/3, 1/2, 1/6 on average for mononucleated cell tissues, skin or bone and viscera, respectively. To apply kP to the whole body, an average value of Y=1/2 can be taken. Experimental observations on tissue protein synthesis and breakdown rates yield direct estimates of kP in satisfactory agreement with comparable theoretical predictions. PMID- 16469139 TI - Dietary inulin intake and age can significantly affect absorption of the faecal marker dysprosium in rats. AB - It is believed that rare earth elements are not absorbed, and thus they are generally used in some mineral absorption studies as a faecal marker. The aim of the present study was to determine the effect of inulin intake and age on dysprosium (Dy) absorption in rats. Eighty male Wistar rats of four different ages (2, 5, 10 and 20 months) were randomised into either a control group or a group receiving 3.75 % inulin in their diet for 4 d and then 7.5 % inulin until the end of the study. The animals were fed fresh food and water ad libitum for 30 d. The intestinal absorption of Dy was determined from a 4 d (day 21 to day 25) balance study. Mean faecal Dy recovery (%) in the eight groups (3 months control, 3 months inulin, 6 months control, 6 months inulin, 11 months control, 11 months inulin, 21 months control, 21 months inulin) was 94.0 (sd 8.6), 64.8 (sd 10.1), 95.8 (sd 9.4), 81.5 (sd 12.1), 98.4 (sd 9.8), 87.8 (sd 9.5), 97.8 (sd 6.2) and 84.9 (sd 10.9), respectively. Our results showed clearly that dietary inulin intake decreased faecal Dy recovery in all four rat groups, and faecal Dy recovery was significantly higher in the old rats (10 and 20 months) than in the young and adult rats. These results show that the faecal recovery (or intestinal absorption) of Dy may vary greatly with nutritional or physiological states such as inulin intake or age. The use of rare earth elements as a faecal marker should be thus validated under each nutritional or physiological state before being employed in mineral absorption studies. PMID- 16469140 TI - Menadione is a metabolite of oral vitamin K. AB - Phylloquinone is converted into menaquinone-4 and accumulates in extrahepatic tissues. Neither the route nor the function of the conversion is known. One possible metabolic route might be the release of menadione from phylloquinone by catabolic activity. In the present study we explored the presence of menadione in urine and the effect of vitamin K intake on its excretion. Menadione in urine was analysed by HPLC assay with fluorescence detection. Urine from healthy male volunteers was collected before and after administration of a single dose of K vitamins. Basal menadione excretion in non-supplemented subjects (n 6) was 5.4 (sd 3.2) microg/d. Urinary menadione excretion increased greatly after oral intake of the K vitamins, phylloquinone and menaquinone-4 and -7. This effect was apparent within 1-2 h and peaked at about 3 h after intake. Amounts of menadione excreted in 24 h after vitamin K intake ranged, on a molar basis, from 1 to 5 % of the administered dose, indicating that about 5-25 % of the ingested K vitamins had been catabolized to menadione. Menadione excretion was not enhanced by phylloquinone administered subcutaneously or by 2',3'-dihydrophylloquinone administered orally. In archived samples from a depletion/repletion study (Booth et al. (2001) Am J Clin Nutr 74, 783-790), urinary menadione excretion mirrored dietary phylloquinone intake. The present study shows that menadione is a catabolic product of K vitamins formed after oral intake. The rapid appearance in urine after oral but not subcutaneous administration suggests that catabolism occurs during intestinal absorption. The observations make it likely that part of the menaquinone-4 in tissues results from uptake and prenylation of circulating menadione. PMID- 16469141 TI - Interaction of fish oil and a glucocorticoid on metabolic responses to an oral glucose load in healthy human subjects. AB - Compared with saturated fat, n-3 long-chain PUFA-rich fish oil improves insulin sensitivity in rats. We studied whether n-3 long-chain PUFA could prevent insulin resistance induced by dexamethasone (a glucocorticoid) in healthy human volunteers. A group of eight subjects was studied twice after a 2 d dexamethasone treatment, before and after a 3-week supplementation with fish oil (providing daily doses of 1.1 g 20 : 5n-3 and 0.7 g 22 : 6n-3). The subjects were studied during the basal state and over the 6 h following an oral glucose load (1 g/kg). Plasma glucose fluxes were traced with [6,6-2H2]glucose and [13C]glucose (naturally 13C-enriched corn glucose). Substrate oxidation was obtained from indirect calorimetry. Following fish oil supplementation, plasma glucose fluxes and substrate oxidation were maintained despite a 17 % reduction (P<0.05) in the area under the curve of plasma insulin response, suggesting an insulin sensitizing effect. PMID- 16469142 TI - Time course of the development of non-alcoholic hepatic steatosis in response to high-fat diet-induced obesity in rats. AB - The aim of the study was to characterize the time course of the development of high-fat diet-induced hepatic steatosis and its relation to body fat accretion and changes in plasma lipid profile. Female Sprague-Dawley rats were high-fat fed (HF; 42 %, kJ) for 1, 2, 4, 6, 12 and 16 weeks and compared to standard fed rats (SD). Data obtained from HF rats were further analysed by classifying the animals into obesity-prone and obesity-resistant. In HF rats, liver lipid content increased rapidly by approximately 200 % during the first 2 weeks, decreased almost to baseline levels between weeks 2 and 6, and re-increased by 17 % between weeks 6 and 16 (P<0.05). Body weight, body fat accretion, plasma leptin, NEFA and glycerol concentrations were higher in HF than in SD rats (P<0.05). These higher values were established in 2 weeks and the differences between the groups did not further enlarge from weeks 2 to 16. Obesity-prone rats depicted higher body weight and body fat accretion than obesity-resistant and SD rats. Surprisingly, however, liver lipid content was the same in obesity-prone as in obesity resistant rats as they were both higher than in SD rats (weeks 2 and 16; P<0.05). Our data support the hypothesis that the liver acts as a systemic buffer, largely increasing its lipid content in the early stage of high-fat feeding. Our results also suggest that the development of non-alcoholic hepatic steatosis is more linked to dietary fat ingestion than to body weight gain. PMID- 16469143 TI - Kinetics of the appearance of cereal alkylresorcinols in pig plasma. AB - Alkylresorcinols (AR) are phenolic lipids found in high amounts in wholegrain wheat and rye. They may be important as bioactive components and/or biomarkers of wholegrain wheat and rye intake. AR absorption was studied with pigs during a habitual diet and after a single meal of AR-rich diet. In study 1, four pigs were fed wholegrain rye (RD) and white wheat bread (WD) for 1 week in a cross-over design. Arterial and venous AR concentrations were determined over 540 min after feeding on days 5 and 7. In study 2, four pigs were fed a synthetic, starch-based diet for 5 d, and arterial and venous AR concentrations were determined over 960 min after feeding a single meal of RD. In study 1, plasma AR concentrations after consuming RD peaked at 120 min, remained elevated between 120 and 420 min, and were re-elevated on refeeding at 480 min. In study 2, AR appeared in the plasma after 60 min, peaked at 180-240 min, and remained elevated after 960 min. The apparent elimination half-life was 3.93 h, and the maximum plasma concentration was 666 (sem 35) nmol/l. Arterial and venous plasma AR concentrations were similar, suggesting that AR are absorbed via the lymphatic system. AR concentrations in pig plasma increase rapidly after a meal containing AR, and persist in plasma for at least 5 d, indicating that AR may be a useful biomarker of wholegrain wheat and rye intake. PMID- 16469144 TI - Meal-induced insulin sensitization in conscious and anaesthetized rat models comparing liquid mixed meal with glucose and sucrose. AB - We have recently shown that meal-induced insulin sensitization (MIS) occurs after feeding and decreases progressively to insignificance after 24 h of fasting and is caused by action of a hepatic insulin sensitizing substance (HISS). In order to carry out quantitative studies of MIS, some standardized meal intake is required. Our objective was to establish animal models to be tested in both the conscious and anaesthetized state using intragastric injection of liquid meals in order to quantify MIS. Insulin sensitivity was assessed before and 90 min after the meal using the rapid insulin sensitivity test (RIST) which is a transient euglycaemic clamp. Rats tested in the conscious state were instrumented under anaesthesia 6-9 d prior to testing with catheters in the carotid artery, jugular vein and stomach. Meals, injected into the stomach, consisted of a liquid mixed meal, sucrose, glucose or water. The glucose sequestration in response to insulin increased by 90 % and 61 % following the liquid mixed meal (10 ml/kg) in conscious and anaesthetized rats, respectively. Glucose, sucrose and water did not effectively activate MIS. MIS was completely reversed in the conscious model by atropine and completely prevented from developing in the anaesthetized model that had previously undergone hepatic denervation. Gastric administration of a liquid mixed meal but not glucose or sucrose is capable of activating MIS for purposes of mechanistic studies and quantification of the MIS process. The feeding signal is mediated by the hepatic parasympathetic nerves. PMID- 16469145 TI - Activity pattern and energy expenditure due to physical activity before and during pregnancy in healthy Swedish women. AB - Human pregnancy is associated with increased requirements for dietary energy and this increase may be partly offset by reductions in physical activity during gestation. Studies in well-nourished women have shown that the physical activity level (PAL), obtained as the total energy expenditure (TEE) divided by the BMR, decreases in late pregnancy. However, it is not known if this decrease is really caused by reductions in physical activity or if it is the result of decreases in energy expenditure/BMR (the so-called metabolic equivalent, MET) for many activities in late pregnancy. In the present study activity pattern, TEE and BMR were assessed in twenty-three healthy Swedish women before pregnancy as well as in gestational weeks 14 and 32. Activity pattern was assessed using a questionnaire and heart rate recording. TEE was assessed using the doubly labelled water method and BMR was measured by means of indirect calorimetry. When compared to the pre-pregnant value, there was little change in the PAL in gestational week 14 but it was significantly reduced in gestational week 32. Results obtained by means of the questionnaire and by heart rate recording showed that the activity pattern was largely unaffected by pregnancy. The findings support the following conclusion: in a population of well-nourished women where the activity pattern is maintained during pregnancy, the increase in BMR represents approximately the main part of the pregnancy-induced increase in TEE, at least until gestational week 32. PMID- 16469147 TI - Serum selenium determinants in French adults: the SU.VI.M.AX study. AB - The objective of the present work was to assess the relationship between serum Se concentrations and environmental determinants (i.e. lifestyle, social activity, geographic region, urban status, education, familial status, physical activity, BMI, tobacco, and food and alcohol consumption). Baseline results from 13 017 subjects (7876 women aged 35-60 and 5141 men aged 45-60) who participated in the SU.VI.M.AX (Supplementation en Vitamines et Mineraux Antioxydants) study were analysed. Fewer than 2 % of the volunteers had a serum Se status under 0.75 micromol/l, which has been quoted as the cut-off of biological Se sub-deficiency. Women had significantly lower serum Se concentrations than men (1.09 (sd 0.19) micromol/l (n 7423) and 1.14 (sd 0.20) micromol/l (n 4915), P<0.0001, respectively). Significant differences in serum Se concentrations were observed between geographic areas. In both sexes, the serum Se concentration increased with alcohol, meat and fish consumption, and decreased with smoking. In premenopausal women, the serum Se concentration was higher in contraceptive-pill users than in non-users. In women only, age was associated with increased serum Se concentrations, and obesity (BMI> or =30 kg/m2) was associated with decreased serum Se levels. In men, we observed a decrease in serum Se concentrations with increased consumption of vegetables and fruits. In conclusion, though few of the volunteers participating in the SU.VI.M.AX study had Se status in the sub deficiency range, 83 % of women and 75 % of men had serum concentrations below the value considered optimal for glutathione peroxidase activity. The largest Se associations in both sexes were found with regions, smoking, alcohol, meat and fish consumption. Further studies are needed to understand the difference in Se status between genders. PMID- 16469146 TI - Improvement of the human intestinal flora by ingestion of the probiotic strain Lactobacillus johnsonii La1. AB - To exert beneficial effects for the host, for example, improving the intestinal microflora, a probiotic must reach the intestine as a viable strain. These properties must be demonstrated by in vitro as well as in vivo methods. However, only a few well-designed human clinical studies have shown these properties. Lactobacillus johnsonii La1 has been shown to give many beneficial effects for the host, but it is unclear whether a viable strain of L. johnsonii La1 has the effect of improving host intestinal microflora. In the present study, a randomised double-blind placebo-controlled cross-over trial was conducted to elucidate the effect of L. johnsonii La1 on human intestinal microflora. Twenty two young healthy Japanese women were randomly divided into two groups, and either received fermented milk with L. johnsonii La1 or a fermented milk without L. johnsonii La1 (placebo) daily for 21 d. Consumption of the fermented milk: (a) increased total Bifidobacterium and Lactobacillus, and decreased lecithinase positive Clostridium in the faeces; (b) increased the faecal lactic acid concentrations; (c) decreased the faecal pH; (d) increased the defecation frequency. These changes were stronger than those observed with the placebo. L. johnsonii La1 was identified in all subjects only after the consumption of the fermented milk. These results suggest that L. johnsonii La1 can contribute to improve intestinal microflora with probiotic properties. PMID- 16469148 TI - The nutritional status of children with cystic fibrosis. AB - The importance of nutritional intervention for children with cystic fibrosis (CF) is well recognised. It would be expected that the increase in knowledge over the past decade would be reflected in improvements in nutritional status for the CF paediatric population. The aim of the present paper was to evaluate the nutritional status of children with CF, cross-sectionally and longitudinally. Body cell mass adjusted for gender and size (BCM/Htp) was measured in sixty-four children with CF to represent nutritional status and expressed as a Z-score. The cross-sectional results showed a mean BCM/Htp Z-score of 0.54 (sd 1.21), with males having a slightly higher Z-score than females but with a larger variation. At the initial measurement, only one female and one male were considered sub optimally nourished. The longitudinal analysis after 2 years showed that the mean population had a significantly decreased BCM/Htp Z-score; however, when each gender was analysed separately, this decrease was significant only in the males. At the final measurement, only two females and three males were considered sub optimally nourished. It is evident from our results that children with CF are well nourished, with only a small percentage considered malnourished. It appears that nutritional status decreases with age, with this decline being more evident in males. These results signify that although children with CF are better nourished with current treatment support, intervention needs to continue throughout a CF patient's life to counteract the changes that occur with age. PMID- 16469149 TI - 'Malnutrition Universal Screening Tool' predicts mortality and length of hospital stay in acutely ill elderly. AB - Malnutrition and its impact on clinical outcome may be underestimated in hospitalised elderly as many screening procedures require measurements of weight and height that cannot often be undertaken in sick elderly patients. The 'Malnutrition Universal Screening Tool' ('MUST') has been developed to screen all adults, even if weight and/or height cannot be measured, enabling more complete information on malnutrition prevalence and its impact on clinical outcome to be obtained. In the present study, 150 consecutively admitted elderly patients (age 85 (sd 5.5) years) were recruited prospectively, screened with 'MUST' and clinical outcome recorded. Although only 56 % of patients could be weighed, all (n 150) could be screened with 'MUST'; 58 % were at malnutrition risk and these individuals had greater mortality (in-hospital and post-discharge, P<0.01) and longer hospital stays (P=0.02) than those at low risk. Both 'MUST' categorisation and component scores (BMI, weight loss, acute disease) were significantly related to mortality (P<0.03). Those patients with no measured or recalled weight ('MUST' subjective criteria used) had a greater risk of malnutrition (P=0.01) and a poorer clinical outcome (P<0.002) than those who could be weighed and, within both groups, clinical outcome was worse in those at risk of malnutrition. The present study suggests that 'MUST' predicts clinical outcome in hospitalised elderly, in whom malnutrition is common (58 %). In those who cannot be weighed, a higher prevalence of malnutrition and associated poorer clinical outcome supports the importance of routine screening with a tool, like 'MUST', that can be used to screen all patients. PMID- 16469150 TI - Magnetic resonance imaging of the behaviour of oil-in-water emulsions in the gastric lumen of man. AB - Pre-processed foods often contain a high percentage of lipid, present as emulsions stabilised with various surface-active agents. The acidic gastric environment can affect the behaviour of such emulsions, modifying the lipid spatial distribution and, in turn, the rate of gastric emptying and nutrient delivery to the gut. The aim of the present study was to use echo-planar magnetic resonance imaging (EPI) to determine the behaviour of model olive oil emulsions during gastric processing. Six healthy male volunteers were intubated nasogastrically on two separate occasions and fed 500 ml 15 % (w/w) olive oil-in water, surfactant-stabilised emulsions designed to have identical droplet size distribution and which were either stable or unstable under gastric acid conditions. EPI was used to assess the oil fraction of the intragastric emulsions, gastric emptying and to visualise the spatial distribution of the oil at 10, 30 and 50 min postprandially. The in vivo imaging measurements of the oil volume fraction of the emulsions correlated well (r 0.66, acid-stable; r 0.52, acid-unstable) with that assayed in the gastric aspirates. Compared with the acid stable emulsion, the acid-unstable emulsion in the gastric lumen rapidly separated into lipid-depleted 'aqueous' and lipid layers. Phase separation in the acid-unstable meal allowed the oil-depleted component to empty first and more rapidly than the stable emulsion as determined by the gastric emptying curves. These pilot data suggest that gastric processing and emptying of high-fat foods could be manipulated by careful choice of emulsifier. PMID- 16469151 TI - The hypotensive response to oral fat is comparable but slower compared with carbohydrate in healthy elderly subjects. AB - The objective of the present study was to determine the comparative hypotensive responses to drinks containing predominantly fat and carbohydrate (CHO) in healthy elderly subjects. Using a randomised, cross-over study, the participants, twelve elderly subjects, six of them female (72.2 (sd 5.7) years), were investigated. On three separate days, blood pressure (BP) and heart rate were measured following ingestion of 300 ml drinks containing: (1) CHO (75 g glucose and 93 g Polyjoule (CHO polymer) providing 2732 kJ (653 kcal)); (2) 88 % fat (cream blended with milk providing 2732 kJ (653 kcal)); (3) water. Systolic BP decreased following the CHO drink (P<0.001) and the high-fat drink (P<0.001) but not water; there was no difference in the magnitude of the decrease between the CHO drink and the drink containing fat (13.4 v. 15.6 mmHg). However, the onset of the fall was slower after the fat-containing drink (13.0 v. 26.5 min (P=0.01); area under the curve for 0-30 min for CHO drink -6.5 v. fat-containing drink 125.4 mmHg x min (P=0.043)). We conclude that ingestion of a high-fat drink results in a comparable fall in BP to a CHO drink although the onset is relatively slower. These observations may have implications for the management of postprandial hypotension. PMID- 16469152 TI - Beef conjugated linoleic acid isomers reduce human cancer cell growth even when associated with other beef fatty acids. AB - Although many data are available concerning anticarcinogenic effects of industrial conjugated linoleic acid (CLA), few studies have reported the antitumour properties of CLA mixtures originating from ruminant products. The aim of the present study was to investigate the in vitro antiproliferative effects of beef CLA mixtures on breast, lung, colon, melanoma and ovarian human cancer cell lines. For this purpose, four fatty acid (FA) extracts prepared from beef lipid and varying in their CLA composition, their corresponding purified CLA-enriched fractions, and mixtures of pure synthetic CLA, the composition of which reproduced that of the four selected beef samples, were tested on cancer cell lines. Cancer cells were exposed for 48 h to medium containing 100 microm-FA and their proliferation was determined by quantifying cellular DNA content (Hoechst 33342 dye). Compared with cells incubated without FA, the number of cancer cells was reduced from 25 to 67 % (P<0.0001) following FA treatment. Antiproliferative effects of CLA mixtures varied in magnitude according to the source of FA, the CLA composition and the cell lines. CLA mixtures naturally present in beef inhibited the proliferation of human cancer cell lines, a high content in cis trans isomers allowing the most important antiproliferative effect. Beef total FA exhibited a greater growth-inhibitory activity than their corresponding CLA enriched fractions. These results suggested that either beef FA other than beef CLA could possess antiproliferative properties and/or the existence of complementary effects of non-conjugated FA and CLA, which could favour the antiproliferative properties of beef total FA. PMID- 16469153 TI - Body composition changes in haemodialysis patients with secondary hyperparathyroidism after parathyroidectomy measured by conventional and vector bioimpedance analysis. AB - Considering the negative effects of secondary hyperparathyroidism (SHPT) in patients with chronic renal failure (CRF), the objective of the present study was to evaluate body composition changes using conventional and vector bioimpedance analysis in patients before and after parathyroidectomy (PTX). Twelve adult patients, mean age 43.4 (sd 12.7) years, were evaluated prior to and 6 months after PTX. Diets were assessed with 3 d dietary records, and mean energy, protein, calcium and phosphorus intake were estimated from these inventories. Weight, height, BMI and bioelectrical impedance were measured; and biochemical markers of nutritional status (albumin and total protein) and bone metabolism (calcium, phosphorus and intact parathyroid hormone) were determined. No significant differences were observed in mean energy, protein and phosphorus after surgery. There was a significant increase in calcium intake after PTX (382.3 (sd 209.6) mg to 656.6 (sd 313.8) mg; P<0.05). Mean weight, BMI, conventional bioelectrical impedance measurements, total body fat, lean body mass and total body water were unaffected by surgery. However, the phase angle and reactance significantly increased after PTX (5.0 degrees (sd 1.4) to 5.6 degrees (sd 1.3); 44.1 (sd 15.6) Omega to 57.1 (sd 14.4) Omega, respectively). The high levels of intact parathyroid hormone before surgery had a negative effect on total body fat (r -0.69, P<0.05). After PTX, the mean albumin significantly increased (3.9 (sd 0.4) g/dl to 4.2 (sd 0.6) g/dl; P<0.05). PTX for SHPT is associated with certain changes in laboratory values, dietary intake and body composition. The latter is best seen with bioimpedance vector analysis. PMID- 16469154 TI - Longitudinal associations between body mass index and serum carotenoids: the CARDIA study. AB - Cross-sectional studies report an inverse association between BMI and serum carotenoid concentration. The present study examined the prospective association between BMI and the serum concentration of five carotenoids in the Coronary Artery Risk Development in Young Adults (CARDIA) study. Serum carotenoids (alpha carotene, beta-carotene, beta-cryptoxanthin, zeaxanthin/lutein, lycopene), BMI, dietary intake, physical activity and dietary supplement use were measured at years 0 and 7 in 3071 black and white male and female participants, who were either persistent smokers or non-smokers. Among non-smokers, year 0 BMI predicted year 7 serum carotenoid levels: obese subjects (BMI > or =30 kg/m2) had an average concentration of the sum of four carotenoids (alpha-carotene +beta carotene + zeaxanthin/lutein+beta-cryptoxanthin) that was 22 % lower than the concentration among subjects with a BMI of less than 22 kg/m2. In contrast, the sum of carotenoids among smokers was only 6 % lower. Relationships between BMI and serum lycopene were weak. The change from year 0 to year 7 in serum carotenoids, except for lycopene, was inversely associated with the change in BMI among non-smokers but not among smokers. Parallel findings were observed for BMI and serum gamma-glutamyl transferase level. In summary, the observation that BMI predicted the evolution of serum carotenoids during a 7-year follow-up among young non-smoking adults is consistent with the hypothesis that carotenoids are decreased in protecting against oxidative stress generated by adipose tissue, while smokers maintain a minimal level of serum carotenoids independent of adiposity. The results for lycopene were, however, discordant from those of the other carotenoids. PMID- 16469155 TI - Effect of soya protein on serum lipid profile and lipoprotein concentrations in patients undergoing hypercholesterolaemic haemodialysis. AB - Clinical trials have shown that soya protein reduces the concentrations of some atherogenic lipids in subjects with normal renal function. The present study examined the effects of soya protein on serum lipid concentrations and lipoprotein metabolism in patients on hypercholesterolaemic haemodialysis. Twenty six hypercholesterolaemic (total cholesterol > or =6.21 mmol/l) patients on haemodialysis were studied in a randomized, double-blind, placebo-controlled clinical trial. After a 4-week run-in phase, the subjects were randomly assigned to two groups. Isolated soya protein or milk protein 30 g was consumed daily as a beverage at breakfast or post-dialysis for 12 weeks. Soya protein substitution resulted in significant reductions in total cholesterol (17.2 (sd 8.9 )%), LDL cholesterol (15.3 (sd 12.5 )%), apo B (14.6 (sd 12.1 )%) and insulin (23.8 (sd 18.7) %) concentrations. There were no significant changes in HDL-cholesterol or apo A-I. These results indicate that replacing part of the daily protein intake with soya protein has a beneficial effect on atherogenic lipids and favourably affects lipoprotein metabolism in hypercholesterolaemic patients undergoing haemodialysis. PMID- 16469156 TI - Lupin kernel fibre foods improve bowel function and beneficially modify some putative faecal risk factors for colon cancer in men. AB - Consumption of some dietary fibres may benefit bowel health; however, the effect of Australian sweet lupin (Lupinus angustifolius) kernel fibre (LKFibre) is unknown. The present study examined the effect of a high-fibre diet containing LKFibre on bowel function and faecal putative risk factors for colon cancer compared to a control diet without LKFibre. Thirty-eight free-living, healthy men consumed an LKFibre and a control diet for 1 month each in a single-blind, randomized, crossover study. Depending on subject energy intake, the LKFibre diet was designed to provide 17-30 g/d fibre (in experimental foods) above that of the control diet. Bowel function self-perception, frequency of defecation, transit time, faecal output, pH and moisture, faecal levels of SCFA and ammonia, and faecal bacterial beta-glucuronidase activity were assessed. In comparison to the control diet, the LKFibre diet increased frequency of defecation by 0.13 events/d (P=0.047), increased faecal output by 21 % (P=0.020) and increased faecal moisture content by 1.6 % units (P=0.027), whilst decreasing transit time by 17 % (P=0.012) and decreasing faecal pH by 0.26 units (P<0.001). Faecal butyrate concentration was increased by 16 % (P=0.006), butyrate output was increased by 40 % (P=0.002) and beta-glucuronidase activity was lowered by 1.4 micromol/h per g wet faeces compared to the control diet (P<0.001). Addition of LKFibre to the diet incorporated into food products improved some markers of healthy bowel function and colon cancer risk in men. PMID- 16469157 TI - Intake of micronutrients high in animal-source foods is associated with better growth in rural Kenyan school children. AB - Observational studies have shown that children in developing countries consuming diets containing high amounts of bioavailable nutrients, such as those found in animal-source foods, grow better. The present study investigated which specific nutrients from the diet of Kenyan school children predicted their growth. The children (n 544, median age 7 years) participated in a 2-year long food supplementation study with animal-source foods. Height gain during the intervention period was positively predicted by average daily intakes of energy from animal-source foods, haem Fe, preformed vitamin A, Ca and vitamin B12. Weight gain was positively predicted by average daily intakes of energy from animal-source foods, haem Fe, preformed vitamin A, Ca and vitamin B12. Gain in mid-upper-arm muscle area was positively predicted by average daily intakes of energy from animal-source foods and vitamin B12. Gain in mid-upper-arm fat area was positively predicted by average daily intakes of energy from animal-source foods. Gain in subscapular skinfold thickness was not predicted by any of the nutrient intakes. Negative predictors of growth were total energy and nutrients that are contained in high amounts in plant foods. The study shows that growth was positively predicted by energy and nutrients that are provided in high amounts and in a bioavailable form in meat and milk, and their inclusion into the diets of children in developing countries should be part of all food-based programmes in order to improve micronutrient status and growth. PMID- 16469158 TI - Hyperglycaemia and reduced glucokinase expression in weanling offspring from dams maintained on a high-fat diet. AB - High-fat feeding reduces the expression of GLUT-2 and the glycolytic enzyme glucokinase (GK). The transcription factor, pancreatic duodenal homeobox-1 (Pdx 1), is important for beta-cell maintenance. The aim of the present study was to determine, in weanling Wistar rats, the effect of a maternal high-fat diet (HFD) during defined periods of gestation and lactation, on body weight, circulating glucose and insulin concentrations, and the expression of GLUT-2, GK and Pdx-1. At postnatal day 21, weights were recorded and glucose and insulin concentrations were measured. The expression levels for mRNA were quantified by LightCycler PCR. Pancreatic sections, immunostained for GLUT-2, GK or Pdx-1, were assessed by image analysis. Weanlings from dams fed an HFD throughout gestation were lighter, with heavier weanlings produced from dams fed an HFD throughout gestation and lactation. Both these groups of weanlings were normoglycaemic, all the others being hyperglycaemic. Hypoinsulinaemia was evident in weanlings from dams fed an HFD throughout gestation only and also for either the first week of lactation or throughout lactation. GLUT-2 mRNA expression was reduced and GLUT-2 immunoreactivity was increased in most of the weanlings. GK mRNA expression and immunoreactivity was reduced in most of the offspring. Pdx-1 mRNA expression was increased in weanlings from dams fed an HFD throughout both gestation and lactation and reduced in those from dams only fed a lactational HFD. Normal Pdx-1 immunoreactivity was found in all of the weanlings. A maternal HFD induces hyperglycaemia in weanlings concomitant with reduced GK expression which may compromise beta-cell function. PMID- 16469159 TI - Towards understanding of glycaemic index and glycaemic load in habitual diet: associations with measures of glycaemia in the Insulin Resistance Atherosclerosis Study. AB - Epidemiologic studies have applied the glycaemic index (GI) and glycaemic load (GL) to assessments of usual dietary intake. Results have been inconsistent particularly for the association of GI or GL with diabetes incidence. We aimed to advance understanding of the GI and GL as applied to food frequency questionnaires (FFQ) by evaluating GI and GL in relation to plasma measures of glycaemia. Included were 1255 adults at a baseline examination (1994-6) and 813 who returned for the 5-year follow-up examination. Usual diet, at both examinations, was assessed by a validated FFQ. GI and GL were evaluated in relation to average fasting glucose (two measures at each examination) and 2 h post-75 g glucose load plasma glucose (baseline and follow-up), and glycated haemoglobin (A1c; follow-up only); using generalized linear models. Correlation coefficients (r) for GI and GL related to measures of glycaemia, adjusted for total energy intake, ranged from -0.004 to 0.04 (all NS) for both examinations. Adjustment for potential confounders, for fasting glucose in models for 2 h glucose (to model incremental glucose) and for average fasting glucose in models for A1c (to account, in part, for overnight endogenous glucose production) also did not materially alter findings, nor did inclusion of data from both examinations together in linear mixed models. The present results call into question the utility of GI and GL to reflect glycaemic response to food adequately, when used in the context of usual diet. Further work is needed to quantify usual dietary exposures relative to glucose excursion and associated chronic glycaemia and other metabolic parameters. PMID- 16469160 TI - Soya phytonutrients act on a panel of genes implicated with BRCA1 and BRCA2 oncosuppressors in human breast cell lines. AB - Breast cancer is the most common cancer in women and a significant cause of death. Mutations of the oncosuppressor genes BRCA1 and BRCA2 are associated with a hereditary risk of breast cancer, and dysregulation of their expression has been observed in sporadic cases. Soya isoflavones have been shown to inhibit breast cancer in studies in vitro, but associations between the consumption of isoflavone-containing foods and breast cancer risk have varied in epidemiological studies. Soya is a unique source of the phytoestrogens daidzein (4',7 dihydroxyisoflavone) and genistein (4',5,7-trihydroxyisoflavone), two molecules that are able to inhibit the proliferation of human breast cancer cells in vitro. The aim of the present study was to determine the effects of genistein (5 microg/ml) and daidzein (20 microg/ml) on transcription in three human breast cell lines (one dystrophic, MCF10a, and two malignant, MCF-7 and MDA-MB-231) after 72 h treatment. The different genes involved in the BRCA1 and BRCA2 pathways (GADD45A, BARD1, JUN, BAX, RB1, ERalpha, ERbeta, BAP1, TNFalpha, p53, p21Waf1/Cip1, p300, RAD51, pS2, Ki-67) were quantified by real-time quantitative RT-PCR, using the TaqMan method and an ABI Prism 7700 Sequence Detector (Applied Biosystems). We observed that, in response to treatment, many of these genes were overexpressed in the breast cancer cell lines (MCF-7 and MDA-MB-231) but not in the dystrophic cell line (MCF10a). PMID- 16469161 TI - The facilitative effects of glucose ingestion on memory retrieval in younger and older adults: is task difficulty or task domain critical? AB - The ingestion of a glucose-containing drink has been shown to improve cognitive performance, particularly memory functioning. However, it remains unclear as to the extent to which task domain and task difficulty moderate the glucose enhancement effect. The aim of this research was to determine whether boosts in performance are restricted to particular classes of memory (episodic v. semantic) or to tasks of considerable cognitive load. A repeated measures (25 g glucose v. saccharin), counterbalanced, double-blind design was used with younger and older adults. Participants performed a battery of episodic (e.g. paired associate learning) and semantic memory (e.g. category verification) tasks under low and high cognitive load. Electrophysiological measures (heart rate and galvanic skin response) of arousal and mental effort were also gathered. The results indicated that whilst glucose appeared to aid episodic remembering, cognitive load did not exaggerate the facilitative effect. For semantic memory, there was little evidence to suggest that glucose can boost semantic memory retrieval even when the load was manipulated. One exception was that glucose facilitated performance during the difficult category fluency task. Regardless, the present findings are consistent with the domain-specific account in which glucose acts primarily on the hippocampal region, which is known to support episodic memory. The possible contribution of the hippocampus in semantic memory processing is also discussed. PMID- 16469162 TI - Effects of orally administered Lactobacillus casei DN-114 001 on the composition or activities of the dominant faecal microbiota in healthy humans. AB - The composition and activities of the faecal microbiota in twelve healthy subjects analysed in a single open study were monitored before (1-week baseline step), during (10 d supplementation step) and after (10 d follow-up step) the ingestion of a fermented milk containing Lactobacillus casei DN-114 001. Fluorescent in situ hybridisation with group-specific DNA probes, real-time PCR using L. paracasei group-specific primers and temporal temperature gradient gel electrophoresis (TTGE) using group-specific primers were carried out, together with bacterial enzyme activity and metabolite analyses to monitor the structure and activities of the faecal microbiota. L. casei DNA was detected in the faeces of all of the subjects by TTGE after 10 d supplementation. Its quantification by real-time PCR showed a 1000-fold increase during the test step compared with initial levels. No major modification in either the dominant members of the faecal microbiota or their activities was observed during the trial. In conclusion, the short-term consumption of a milk product containing L. casei DN 114 001 was accompanied by a high, transient increase in the quantity of this strain in the faeces of all of the subjects without markedly affecting biochemical or bacteriological factors. PMID- 16469163 TI - Oral immunoadjuvant activity of Lactobacillus casei subsp. casei in dextran-fed layer chickens. AB - We recently reported that synbiotic Lactobacillus casei subsp. casei together with specific substrate dextran elicited an enhancement in humoral immune response against bovine serum albumin (BSA) as a model antigen in BALB/c mice. The present study was designed to evaluate the oral immunoadjuvant effects of the synbiotic in layer chickens. Using a PCR assay, L. casei subsp. casei was detected specifically in the intestinal chyme of chickens (10 d of age, Julia strain) fed ad libitum on a diet supplemented with 75 mg dextran/kg (dextran supplemented diet, DSD) and administered orally with 10(7) colony-forming units (CFU) L. casei subsp. casei in 0.1 ml PBS with the aid of an intubation needle at 1, 2 and 3 d of age. Furthermore, oral administration of 10(7) CFU L. casei subsp. casei at 1-3 d of age significantly enhanced the production of anti-BSA antibody in DSD-fed chickens (60 d of age) administered orally with 1 mg BSA at 32 and 33 d of age and subcutaneously with 5 microg BSA at 33 d of age. In addition, among bacterial numbers tested, 10(6) CFU L. casei subsp. casei together with dextran induced an effective increase in humoral immune response to mixed inactivated vaccines against Newcastle disease and avian infectious bronchitis, and the treatment may be advantageous in protecting against these infectious diseases in chickens in actual application. These results suggest that dietary supplementation of L. casei subsp. casei with dextran leads to immunomodulation of humoral immune responses. PMID- 16469164 TI - Mechanisms involved in the antiplatelet effect of C-phycocyanin. AB - C-phycocyanin (cpc), a biliprotein isolated from Spirulina platensis, has been reported to exert many therapeutic and nutritional values. In the present study, we examined whether cpc has an antiplatelet activity in vitro and further investigated the possible anti-aggregatory mechanisms involved. Our results showed that preincubation of cpc (1-50 microg/ml) with rabbit washed platelets dose-dependently inhibited the platelet aggregation induced by collagen (10 microg/ml) or arachidonic acid (100 microm), with an IC50 of about 10 microg/ml. Furthermore, the thromboxane B2 formation caused by collagen or arachidonic acid was significantly inhibited by cpc due to suppression of cyclooxygenase and thromboxane synthase activity. Similarly, the rise of platelet intracellular calcium level stimulated by arachidonic acid and collagen-induced platelet membrane surface glycoprotein IIb/IIIa expression were also attenuated by cpc. In addition, cpc itself significantly increased the platelet membrane fluidity and the cyclic AMP level through inhibiting cyclic AMP phosphodiesterase activity. These findings strongly demonstrate that cpc is an inhibitor of platelet aggregation, which may be associated with mechanisms including inhibition of thromboxane A2 formation, intracellular calcium mobilization and platelet surface glycoprotein IIb/IIIa expression accompanied by increasing cyclic AMP formation and platelet membrane fluidity. PMID- 16469165 TI - Helminth fauna of Talpa spp. in the Palaearctic Realm. AB - The helminth fauna of the genus Talpa in the Palaearctic Realm is reviewed. Several helminth species reported in Talpa spp. by a number of authors are discussed, with reference to host specificity, parasite biology, and host ethology, ecology and phylogeny. Twelve species of cestodes were found, two of which exhibit stenoxenous specificity (Staphylocystis bacillaris and Multitesticulata filamentosa). Only three species of trematodes, Ityogonimus lorum, Ityogonimus ocreatus and Combesia macrobursata, are exclusive parasites of Talpa spp. The largest group are nematodes, with 37 species. Species of Tricholinstowia are parasites of holarctic talpids and several species of distinct genera, such as Capillaria, Soboliphyme and Trichuris, are found only in Talpa spp. Only acanthocephalans of the genus Moniliformis have been reported in moles of the genus Talpa. On the basis of these helminthological findings, the close phylogenetic relationship between moles (Talpidae) and shrews (Soricidae) supports the separation of the ordinal levels Soricomorpha and Erinaceomorpha. PMID- 16469166 TI - Comparative study on DNA sequences of ribosomal DNA and cytochrome c oxidase subunit 1 of mitochondrial DNA among five species of gnathostomes. AB - The nucleotide sequences of partial 18S, complete internal transcribed spacer region 1 (ITS1), complete 5.8S, complete ITS2 and partial 28S of ribosomal DNA (rDNA) and cytochrome c oxidase subunit 1 of mitochondrial DNA (MCOI) from five species of gnathostomes (G. spinigerum, G. doloresi, G. nipponicum, G. hispidum and G. binucleatum with the former four species being distributed in Japan and Asia) that cause human gnathostomiasis were compared by direct polymerase chain reaction cycle-sequencing. The nucleotide sequences of each region of the18S (613 bp), 5.8S (158 bp) and 28S (598 bp) rDNA from the five species were almost identical. The ITS1 region was different in length for the five species. The nucleotide sequences of each region of ITS2 and partial MCO1 regions were different among the five species. Therefore, these two regions can be used as genetic markers for identification of worms. PMID- 16469167 TI - Soil contamination with ascarid eggs in playgrounds in Kirikkale, Turkey. AB - Contamination of soil with feline and canine ascarid eggs in eight playgrounds in Kirikkale, Turkey was investigated monthly from February 2003 to January 2004. Dog faeces were also collected and all samples were examined using the zinc sulphate centrifugal flotation method. Eggs of Toxocara were observed in 5 of 8 (62.5%) of playgrounds examined and in 15.6% of 480 soil samples. The number of eggs varied from 1 to 11. Eggs were observed in soil samples collected in February, March to June, August and November, with embryonated eggs appearing in June and August. Eggs of Toxascaris leonina and Taenia spp. and oocysts of Isospora spp. were also found in 1.5%, 1.0% and 0.2% of soil samples, respectively. Of 26 samples of dog faeces collected, 7.7% were contaminated with Toxocara spp. and 11.5% with Taenia spp. The presence of Toxocara eggs in the city playgrounds and dog populations suggests a potential human health hazard due to toxocariasis. PMID- 16469169 TI - Fertilization state of Ascaris suum determined by electrorotation. AB - Electrorotation is a non-invasive technique that is capable of detecting changes in the morphology and physicochemical properties of microorganisms. The first detailed electrorotation study of the egg (ovum) of a parasitic nematode, namely Ascaris suum is described to show that electrorotation can rapidly differentiate between fertilized and non-fertilized eggs. Support for this conclusion is by optical microscopy of egg morphology, and also from modelling of the electrorotational response. Modelling was used to determine differences in the dielectric properties of the unfertilized and fertilized eggs, and also to investigate specific differences in the spectra of fertilized eggs only, potentially reflecting embryogenesis. The potential of electrorotation as an investigative tool is shown, as undamaged eggs can be subjected to further non destructive and destructive techniques, which could provide further insight into parasite biology and epidemiology. PMID- 16469168 TI - Enzyme-linked immunosorbent assay with worm vomit and cercarial secretions of Schistosoma mansoni to detect infections in an endemic focus of Burkina Faso. AB - Cercariae and adult Schistosoma mansoni were used to prepare, respectively, cercarial secretions (CS) and worm vomit (WoV). These were used as antigens in an enzyme-linked immunosorbent assay (ELISA) to test the IgG-reactivity of sera obtained in an S. mansoni-endemic area of Burkina Faso. Among the egg-excreting individuals (n = 240), 94.6% reacted positively with WoV, but only 62.9% with CS, thus suggesting a high diagnostic sensitivity of WoV, but not of CS. Among those individuals without detectable eggs in two Kato-Katz thick smears from different stool specimens (n = 215), the respective percentages of positive IgG reactivity were 78.1% and 63.3%. These positive reactions in the absence of detectable eggs are interpreted in terms of limited sensitivity of parasitological stool examinations. Optical density values in ELISA with CS, but not with WoV, correlated negatively with age, which may reflect decreasing exposure to cercariae in older individuals. PMID- 16469170 TI - A survey of waterfowl for echinostomes and schistosomes from Lake Wanaka and the Waitaki River watershed, New Zealand. AB - Waterfowl from Lake Wanaka and the Waitaki Lakes watershed of New Zealand's South Island were surveyed to find natural hosts with adult echinostomes and schistosomes to provide sufficient numbers of eggs for laboratory studies. The Canada goose (Branta canadensis) was found to host an echinostome determined to be a New Zealand strain of Echinostoma revolutum. The New Zealand scaup (Aythya novaeseelandia) concurrently hosts three species of echinostomes (E. revolutum, Echinoparyphium cinctum and E. recurvatum) plus two species of avian schistosomes (Trichobilharzia sp. and Dendritobilharzia pulverulenta). In the Canada goose and the New Zealand scaup, adult gravid echinostomes predominated over juveniles. In other waterfowl surveyed, very few echinostomes were found, with juveniles predominating. PMID- 16469171 TI - Influence of season and host age on wild boar parasites in Corsica using indicator species analysis. AB - The indicator value (Ind Val) method which combines measures of fidelity and specificity has been used in a study on wild boar parasites in Corsica during 2001-2003. Because of its resilience to changes in abundance, IndVal is a particularly effective tool for ecological bioindication. The Ind Val method showed how season can influence the occurrence of parasite species in the wild boar, and also identified parasites as bioindicators relative to host age. The randomization test identified five parasite species having a significant indicator value for the season (the ticks, Hyalomma aegyptium and Rhipicephalus sanguineus, the louse, Haematopinus suis and the nematodes Globocephalus urosubulatus and Ascaris suum and two indicator species of an age class (the nematodes G. urosubulatus and Metastrongylus sp.). Data on species composition and infection levels would help improve the monitoring and management of parasitism in Suidae populations. PMID- 16469172 TI - Molecular identification of Anisakis species from Pleuronectiformes off the Portuguese coast. AB - Anisakid nematodes belonging to the Anisakis simplex complex are highly prevalent in several fish species off the coast of Portugal and are an important zoonotic problem in the Iberian Peninsula. Two reproductively isolated sibling species of the Anisakis simplex complex were identified from Pleuronectiformes inhabiting the Portuguese coast using restriction fragment length polymorphism (RFLP). Recombinant genotypes corresponding to presumptive Anisakis simplex sensu stricto and Anisakis pegreffii hybrids were also detected by this technique, as well as the species Anisakis typica. Although 25 species of Pleuronectiformes were investigated, Anisakis spp. larvae were only found in seven: Arnoglossus imperialis, Arnoglossus laterna, Lepidorhombus boscii, Citharus linguatula, Platichthys flesus, Dicologlossa cuneata and Solea senegalensis. The occurrence of hybrids in relatively sedentary fishes such as the Pleuronectiformes suggests that the Portuguese coast may constitute an area of hybridization and, therefore, is of particular interest for the study of the process of hybridization and speciation for these anisakids. PMID- 16469173 TI - Changes in FaRP-like peptide levels during development of eggs from the plant parasitic cyst nematode, Heterodera glycines. AB - The plant-parasitic cyst nematode Heterodera glycines requires a host plant to complete its life cycle, which involves hatching of infective juveniles that parasitize through root entry. A laboratory population of H. glycines grown on soybean, Glycine max, undergoes a sharp increase in maturity between 5 and 6 weeks in culture, as measured by the proportion of eggs containing well developed pre-hatch juveniles (late development eggs) versus eggs without visible juveniles (early development eggs). The median percent of eggs classified as late development, representing all samples taken from 4 to 7 weeks in culture, was 61%. For all samples taken up to 5 weeks, 80% scored below the median. In samples taken after 5 weeks, 15% scored below the median. This shift in population maturity was accompanied by a significant increase (P < 0.01) in the number of hatched juveniles present in each sample. There was also a significant increase (P < 0.02) in amount of FaRP-like peptide detected by specific ELISA. Total FaRP levels increased from 0.18 +/- 0.07 fMol FLRFamide equivalents per ng protein in early development eggs to 0.40 +/- 0.17 in late development eggs. The level remained high in hatched juveniles. HPLC/ELISA detected as many as nine potential FaRPs in H. glycines, two of which were specifically increased (P < 0.005) in hatched juveniles. The association of FaRPs with maturing eggs and the possible involvement of these neuropeptides with juvenile hatching and motility are discussed. PMID- 16469174 TI - Diversity of Dilepididae (Cestoda: Cyclophyllidea) revealed by cytogenetic analysis. AB - Karyotypes of three dilepidid species: Molluscotaenia crassiscolex, Anomotaenia bacilligera and Dilepis undula, which have not been recorded previously, were studied using conventional Giemsa staining and comparative karyometric analysis. Twelve small biarmed chromosomes were observed in mitotic cells of M. crassiscolex, 16 biarmed chromosomes of gradually decreasing size were found in cells of A. bacilligera, while 18 elements were characteristic for D. undula. These data, together with information available in literature, prove the heterogeneity and possible polyphyletic nature of the family Dilepididae. PMID- 16469175 TI - Prevalence of amphistomiasis and its association with Fasciola gigantica infections in Zambian cattle from communal grazing areas. AB - To estimate prevalence of amphistomiasis and its association with bovine Fasciola gigantica infections in Zambia, 709 faeces of cattle presented for slaughter from three major cattle rearing areas of Central, Southern and Western provinces were analysed. The prevalence rate of amphistomes was 51.6%. Egg counts per gram (EPG) of faeces ranged from 0 to 385 with a mean (+/- SEM) of 11.96 +/- 1.07. The origin of the cattle had a significant influence (P < 0.001) on the prevalence rate. Fasciola gigantica infections accounted for 46.7% of the cattle examined. The mean EPG count (+/- SEM) was 6.3 +/- 0.66 with a range of 0 to 223. A total of 34.6% were mixed infections while single Fasciola and amphistome infections represented 12.1% and 17.1%, respectively. Significantly more cattle (63.8%) were infected with either single or both trematode infections (P < 0.0001) than not. Mixed trematode infections were highest in the Southern province (80.0%) while the lowest were recorded in Central province (16.3%). A positive correlation (r(2) = 0.0428) was present in mixed infections. In order to minimize losses, epidemiology and cross-resistance of amphistomiasis and fascioliasis should be studied in different ecological regions of Zambia to formulate efficient control programmes. PMID- 16469176 TI - Assessment of routine inspection methods for porcine cysticercosis in Zambian village pigs. AB - The value of tongue and meat inspection as diagnostic tools for porcine cysticercosis was assessed in 65 Zambian village pigs by comparing the results with carcass dissections. In addition, the intensity of infections, distribution and viability of cysts in infected pigs were measured. Five pigs (7.7%) were positive on tongue examination, while routine meat inspection showed 12 (18.5%) positives. However, carcass dissections detected cysticerci in 31 (47.7%) pigs. The range in number of cysticerci was 1 to 14,662 per carcass. Cysticerci were distributed throughout the carcass with the highest concentration in the heart, tongue and hind legs. In one animal 13 viable cysts were detected only in the brain. Fourteen pigs had more than 100 viable cysts, six between 2 and 100, and four had single cyst infections. Seven animals harboured only calcified cysts. These findings demonstrate the serious shortcomings of routine detection methods for porcine cysticercosis. While the specificity of tongue palpation and meat inspection was 100%, these tests failed to detect the infection in 83.9% and 61.3% of infected pigs, respectively. PMID- 16469178 TI - Plasma etching and ashing: a technique for demonstrating internal structures of helminths using scanning electron microscopy. AB - Plasma etching and ashing for demonstrating the three-dimensional ultrastructure of the internal organs of helminths is described. Adult worms of the cestode Caryophyllaeides fennica were dehydrated through an ethanol series, critical point dried (Polaron E3000) and sputter coated with 60% gold-palladium (Polaron E5100) and glued to a standard scanning electron microscope (SEM) stub positioned as required for ashing. After initial SEM viewing of worm surfaces for orientation, stubs were placed individually in the reactor chamber of a PT7150 plasma etching and ashing machine. Worms were exposed to a radio frequency (RF) potential in a low pressure (0.2 mbar) oxygen atmosphere at room temperature. The oxidation process was controlled by varying the times of exposure to the RF potential between 2 to 30 min, depending on the depth of surface tissue to be removed to expose target organs or tissues. After each exposure the oxidized layer was blown from the surface with compressed air, the specimen sputter coated, and viewed by SEM. The procedure was repeated as necessary, to progressively expose successive layers. Fine details of organs, cells within, and cell contents were revealed. Ashing has the advantage of providing three dimensional images of the arrangement of organs that are impossible to visualize by any other procedure, for example facilitating testes counts in cestodes. Both freshly-fixed and long-term stored helminths can be ashed. Ashing times to obtain the desired results were determined by trial so that some duplicate material was needed. PMID- 16469177 TI - Toxocara canis infections in a pig model: immunological, haematological and blood biochemistry responses. AB - The immunological, haematological and enzymatic responses to the inoculation in pigs of 100,000 embryonated eggs of Toxocara canis were studied. Fifteen females were inoculated and three remained as controls. Haematological values were analysed from day 7 p.i. until day 126 p.i. In the inoculated group, white blood cells were raised on day 14 p.i. and eosinophil values on days 7, 14, 21, 35 and 49 p.i. showing significant differences compared with controls (P < 0.05). Absolute eosinophil counts (per ml) presented two rises, the first on days 7, 14 and 21 p.i. and the second on days 35 and 49 p.i. Blood biochemistry was maintained within normal values. Serological examination by ELISA to determine antibody levels against Toxocara canis L2/L3 excretory-secretory (ES) antigens showed values higher than the positive cut-off (1:32) from day 7 p.i. and until the end of the study on day 126 p.i., presenting two peaks: one on day 28 p.i. and the second covering days 49 to 56 p.i. Western blots of sera of inoculated animals presented, from day 7 p.i., two polypeptide bands of 55 and 70 kDa MW and, from day 56 p.i., an additional band of 120 kDa MW, all of which persisted until the end of the study. Immunological responses were sustained over time. No direct correlation was observed between the rise in eosinophils and antibody titres. To validate the conclusions, more studies are required on the polypeptide bands. PMID- 16469179 TI - Unusual finding of Trichobilharzia sp. in Motacilla alba in the Czech Republic. AB - Adult male worms of Trichobilharzia sp. recovered from a pied wagtail (Motacilla alba) in the Czech Republic were found to belong to a new species of the genus. The finding of Trichobilharzia sp. in a passeriform bird in Europe represents an important discovery, as only anseriform birds have thus far been reported as final hosts of the European Trichobilharzia species. PMID- 16469180 TI - Sodium dodecyl sulphate as a rapid clearing agent for studying the hard parts of monogeneans and nematodes. AB - Hard structures of helminths have often been used for taxonomic identification but are usually not clearly defined when treated with conventional methods such as ammonium picrate-glycerin for monogeneans and glycerin for nematodes. The present study reports a rapid and simple technique to better resolve the hard parts of selected monogeneans and nematodes using 5-10% alkaline sodium dodecyl sulphate (SDS). In comparison with established methods, SDS-treated worms become more transparent. In monogeneans treated with SDS, clear details of the hooks, hook filaments, anchors, bars and the sclerotized copulatory organs could be observed. In SDS-treated nematodes, spicules and ornamentations of the buccal capsules could be clearly seen. PMID- 16469181 TI - Endocannabinoids in the intact retina: 3 H-anandamide uptake, fatty acid amide hydrolase immunoreactivity and hydrolysis of anandamide. AB - There is much evidence for an endocannabinoid system in the retina. However, neither the distribution of endocannabinoid uptake, the regulation of endocannabinoid levels, nor the role of endocannabinoid metabolism have been investigated in the retina. Here we focused on one endocannabinoid, anandamide (AEA), and its major hydrolyzing enzyme, fatty acid amide hydrolase (FAAH), in the goldfish retina. Immunoblots of FAAH immunoreactivity (IR) in goldfish retina, brain and rat retina, and brain homogenates showed a single band at 61 kDa that was blocked by preadsorption with peptide antigen. Specific FAAH IR (blocked by preadsorption) was most prominent over Muller cells and cone inner segments. Weaker label was observed over some amacrine cells, rare cell bodies in the ganglion cell layer, and in four lamina in the inner plexiform layer. FAAH activity assays showed that goldfish-retinal and brain homogenates hydrolyzed AEA at rates comparable to rat brain homogenate, and the hydrolysis was inhibited by methyl arachidonyl fluorophosphonate (MAFP) and N-(4 hydroxyphenyl) arachidonamide (AM404), with IC(50)s of 21 nM and 1.5 microM, respectively. Cellular 3H-AEA uptake in the intact retina was determined by in vitro autoradiography. Silver-grain accumulation at 20 degrees C was most prominent over cone photoreceptors and Muller cells. Uptake was significantly reduced when retinas were incubated at 4 degrees C, or preincubated with 100 nM MAFP or 10 microM AM404. There was no differential effect of blocking conditions on the distribution of silver grains over cones or Muller cells. The codistribution of FAAH IR and 3H-AEA uptake in cones and Muller cells suggests that the bulk clearance of AEA in the retina occurs as a consequence of a concentration gradient created by FAAH activity. We conclude that endocannabinoids are present in the goldfish retina and underlay the electrophysiological effects of cannabinoid ligands previously shown on goldfish cones and bipolar cells. PMID- 16469182 TI - Horizontal cells in the retina of a diurnal rodent, the agouti ( Dasyprocta aguti ). AB - The morphology and distribution of normally placed and displaced A horizontal cells were studied in the retina of a diurnal hystricomorph rodent, the agouti Dasyprocta aguti. Cells were labeled with anti-calbindin immunocytochemistry. Dendritic-field size reaches a minimum in the visual streak, of about 9,000 microm(2), and increases toward the retinal periphery both in the dorsal and ventral regions. There is a dorsoventral asymmetry, with dorsal cells being larger than ventral cells at equal distances from the streak. The peak value for cell density of 281 +/- 28 cells/mm(2) occurs in the center of the visual streak, decreasing toward the dorsal and ventral retinal periphery, paralleling the increase in dendritic-field size. Along the visual streak, the decline in cell density is less pronounced, remaining between 100-200 cells/mm(2) in the temporal and nasal periphery. Displaced horizontal cells are rare and occur in the retinal periphery. They tend to be smaller than normally placed horizontal cells in the ventral region, whilst no systematic difference was observed between the two cell groups in the dorsal region. Mosaic regularity was studied using nearest-neighbor analysis and the Ripley function. When mosaic regularity was determined removing the displaced horizontal cells, there was a slight increase in the conformity ratio, but the bivariate Ripley function indicated some repulsive dependence between the two mosaics. Both results were near the level of significance. A similar analysis performed in the capybara retina, a closely related hystricomorph rodent bearing a higher density of displaced horizontal cells than found in the agouti, suggested spatial independence between the two mosaics, normally placed versus displaced horizontal cells. PMID- 16469183 TI - Morphological characterization of the retinal degeneration in three strains of mice carrying the rd-3 mutation. AB - Retinal development in 3 strains of rd-3/rd-3 mutant mice, previously shown to have different rates of degeneration, was studied using light, electron, and immunofluorescence microscopy. The time course and phenotype of the degeneration as well as details on the mechanism of massive photoreceptor cell loss are compared with other known retinal degenerations in mice. Up until postnatal day (P) 10, the retinas of all three strains (RBF, 4Bnr, In-30) develop similarly to those of pigmented and nonpigmented controls. TUNEL-positive cells appear in the outer nuclear layer (ONL) by P14, and reach a maximum in all three mutant strains around P21. Scattered rods and cones form a loose, monolayered ONL by 8 weeks in the albino RBF strain, by 10 weeks in the albino 4Bnr strain, and by 16 weeks in the pigmented In-30 strain. Though the initial degeneration begins in the central retina, there is no preferred gradient of cell death between central and peripheral photoreceptors. Rods and cones are present at all ages examined. During development, stacks of outer segments (OS) form in all three strains though they never achieve full adult lengths, and often have disorganized, atypical OS. Rod opsin is expressed in the developing OS but is redistributed into plasma membrane as OS degeneration proceeds. Retinal pigment epithelial (RPE) cells of all mutant strains contain packets of phagocytosed OS, and their apical processes associate with the distal ends of the OS. At their synaptic sites, photoreceptor terminals contain ribbons apposed to apparently normal postsynaptic triads. As photoreceptors are lost, Muller cells fill in space in the ONL but they do not appear to undergo significant hypertrophy or migration, though during the degeneration, glial fibrillary acidic protein (GFAP) expression is gradually upregulated. Macrophage-like cells are found frequently in the subretinal space after the onset of photoreceptor apoptosis. As OS disappear, the RPE apical processes revert to simple microvilli. Late in the degeneration, some RPE cells die and neighboring cells appear to flatten as if to maintain confluence. In regions of RPE cell loss that happen to lie above retina where the ONL is gone, cells of the inner nuclear layer (INL), wrapped by Muller cell processes, may front directly on Bruch's membrane. PMID- 16469184 TI - Visual evoked potentials for red-green gratings reversing at different temporal frequencies: asymmetries with respect to isoluminance. AB - Human visual evoked potentials (VEPs) were recorded for abrupt reversals of 2 cycles/deg (c/deg) square-wave gratings combining high red-green contrast with different levels of luminance contrast. Response characteristics--2nd harmonic amplitudes and peak latencies as a function of luminance contrast--were compared for four different reversal rates ranging from 6.25 Hz to 12.5 Hz. At every reversal frequency, the VEP amplitude and latency plots were nonsymmetrical with respect to isoluminance. The amplitude dropped to a minimum within a region of rapid phase change, always at a red-green luminance contrast for which the green color had the higher luminance, at about 40% or 50% Michelson luminance contrast. The rapid phase shift around this contrast suggested a sudden change in the relative impact of VEP generators with different latencies, possibly dominated by parvocellular or magnocellular input. The most prominent VEP waveform through most of the luminance contrast range, P110, is interpreted in terms of a parvo mediated response that is attenuated with increasing reversal frequency. Contrast dependent changes in the P110 amplitude appear to be responsible for the VEP asymmetries reported here. PMID- 16469185 TI - Visual evoked potentials for reversals of red-green gratings with different chromatic contrasts: asymmetries with respect to isoluminance. AB - Human visual evoked potentials (VEPs) were recorded for abrupt 6.25-Hz reversals of 2 c/deg square-wave gratings combining red-green contrast with different levels of luminance contrast. Response characteristics-- amplitudes and peak latencies as a function of luminance contrast--were compared for four different pairs of red-green colors and an isochromatic yellow grating. For each of the red green color pairs, the plots of VEP amplitudes and latencies were nonsymmetrical with respect to isoluminance. The amplitude dropped to a minimum within a region of rapid phase change, at a different contrast for each color pair but always at a luminance contrast for which the greener color had the higher luminance. When the contrast-response curve for each of the four red-green pairs was modeled by a simple |CL - CM| opponency of L- and M-cone contrast using a fixed CL/CM weighting ratio of about two, there was a close correspondence between the contrast giving a null in the modeled response and that giving a minimum in the VEP amplitude. So for the stimulus parameters applied here, the reversal VEP appeared to be dominated by L/M-opponent response contributions for which the signed CL/CM-cone weighting ratio was close to a value of minus two rather than to a value of minus one, which is characteristic of the psychophysical red-green detection mechanism and representative of CL/CM weighting ratios for precortical cells in the parvocellular pathway. PMID- 16469186 TI - Glycine- and GABA-activated inhibitory currents on axon terminals of rabbit cone bipolar cells. AB - Glycine- and GABA-activated currents were examined in the axon terminals of 12 types of rabbit cone bipolar cells. In the superfused retinal slice, a cell was voltage clamped at 0 mV in the presence of cobalt; then glycine or GABA was puffed onto the axon terminal. Types CBa1, CBa2, and a few CBa1-2 cells demonstrated larger glycine-activated currents than GABA-activated ones. However, some OFF cells (CBa2(n), CBa1-2(n), CBa1(w)), most CBa1-2, and most ON cells (CBb3, CBb3-4, CBb3(n), and CBb4) displayed larger GABA-activated currents. The ON cell, CBb5, possessed only a GABA-activated current. The predominance of glycinergic currents in CBa1, CBa2, and a few CBa1-2 cells suggests a major input from the glycinergic AII amacrine cell and thus a key role for these cells in the rod bipolar pathway. Certain OFF cells (most CBa1-2) expressed larger GABA activated currents. All types expressed both GABA(A) and GABAC currents about equally, although most OFF types (CBa1, CB a2(n), CBa1-2, and CBa2(n)) displayed a slightly greater GABA(A) component. PMID- 16469187 TI - Stimulus-dependent correlated firing in directionally selective retinal ganglion cells. AB - Synchronous spiking has been postulated to be a meta-signal in visual cortex and other CNS loci that tags neuronal spike responses to a single entity. In retina, however, synchronized spikes have been postulated to arise via mechanisms that would largely preclude their carrying such a code. One such mechanism is gap junction coupling, in which synchronous spikes would be a by-product of lateral signal sharing. Synchronous spikes have also been postulated to arise from common source inputs to retinal ganglion cells having overlapping receptive fields, and thus code for stimulus location in the overlap area. On-Off directionally selective ganglion cells of the rabbit retina exhibit a highly precise tiling pattern in which gap junction coupling occurs between some neighboring, same preferred-direction cells. Depending on how correlated spikes arise, and for what purpose, one could postulate that synchronized spikes in this system (1) always arise in some subset of same-direction cells because of gap junctions, but never in non-same-preferred-directional cells; (2) never arise in same-directional cells because their receptive fields do not overlap, but arise only in different directional cells whose receptive fields overlap, as a code for location in the overlap region; or (3) arise in a stimulus-dependent manner for both same- and different-preferred-direction cells for a function similar to that postulated for neurons in visual cortex. Simultaneous, extracellular recordings were obtained from neighboring On-Off directionally selective (DS) ganglion cells having the same and different preferred directions in an isolated rabbit retinal preparation. Stimulation by large flashing spots elicited responses from DS ganglion-cell pairs that typically showed little synchronous firing. Movement of extended bars, however, often produced synchronous spikes in cells having similar or orthogonal preferred directions. Surprisingly, correlated firing could occur for the opposite contrast polarity edges of moving stimuli when the leading edge of a sweeping bar excited the receptive field of one cell as its trailing edge stimulated another. Pharmacological manipulations showed that the spike synchronization is enhanced by excitatory cholinergic amacrine-cell inputs, and reduced by inhibitory GABAergic inputs, in a motion-specific manner. One possible interpretation is that this synchronous firing could be a signal to higher centers that the outputs of the two DS ganglion cells should be "bound" together as responding to a contour of a common object. PMID- 16469188 TI - Retinal ganglion cell coding in simulated active vision. AB - The image on the retina is almost never static. Eye, head, and body movements, and externally generated motion create rapid and continual changes in the retinal image ("active vision"). Virtually all vision in animals such as primates, which make saccades as often as 3-4 times/s, is based on information that must be derived from the first few hundred milliseconds after sudden, global changes in the retinal image. These changes may be accompanied by large changes in area mean luminance, as well as higher order image contrast statistics. This study investigated how retinal ganglion cell responses, whose response properties have been typically studied and defined in a stable stimulus regime, are affected by sudden changes in mean luminance that are characteristic of active vision. Specifically, the steady-state responses of retinal ganglion cells to static or moving square-wave grating stimuli were recorded in an isolated, superfused rabbit eyecup preparation and compared to responses after saccade-like changes in luminance. The manner of coding after luminance changes was different for different ganglion cell classes; both suppression and enhancement of responses to patterns following luminance changes were found. Brisk-transient Off cells unambiguously signaled the darkening of the overall image, but were also modulated by the subsequently appearing grating stimulus. Several types of On center cell behavior were observed, ranging from strong suppression of the subsequent response by luminance changes, to strong enhancement. Overall, most ganglion cells distinguished static patterns after a luminance change via differences in their spike discharges nearly as well as before, although there were clear asymmetries between the On and Off pathways. Changes in mean luminance in some ganglion cells, such as On-Off directionally selective ganglion cells, could create large phase shifts in the response to patterned, moving stimuli, although these stimuli were still detected immediately after luminance changes. The results of this study show that the image dynamics of active vision may be a fundamental challenge for the visual system because of strong effects on retinal ganglion cell function. However, rapid extraction of unambiguous information after luminance changes appears to be encoded in differences in the spike discharges in different retinal ganglion cell classes. Asymmetries among ganglion cell classes in sensitivity to luminance changes may provide a basis by which some provide the "context" for interpreting the firing of others. PMID- 16469189 TI - Rod and cone function in coneless mice. AB - Transgenic coneless mice were initially developed to study retinal function in the absence of cones. In coneless mice created by expressing an attenuated diphtheria toxin under the control of flanking sequences from the human L-cone opsin gene, a small number of cones (3-5% of the normal complement) survive in a retina that otherwise appears structurally quite normal. These cones predominantly ( approximately 87% of the total) contain UV-sensitive photopigment. ERG recordings, photoreceptor labeling, and behavioral measurements were conducted on coneless and wild-type mice to better understand how the nature of this alteration in receptor complement impacts vision. Signals from the small residual population of UV cones are readily detected in the flicker ERG where they yield signal amplitudes at saturation that are roughly proportional to the number of surviving cones. Behavioral measurements show that rod-based vision in coneless mice does not differ significantly from that of wild-type mice, nor does their rod system show any evidence of age-related deterioration. Coneless mice are able to make accurate rod-based visual discriminations at light levels well in excess of those required to reach cone threshold in wild-type mice. PMID- 16469190 TI - Characteristics of period doubling in the human cone flicker electroretinogram. AB - Electroretinogram (ERG) responses of the cone system to a flickering stimulus can exhibit a cyclic variation in amplitude. This phenomenon of synchronous period doubling has been attributed to a nonlinear feedback mechanism within the retina that alters response gain. The aim of the present study was to investigate intersubject variability in period doubling in the ERG of the human cone system, and to assess the implications of this variability for signal processing within the retina. Period doubling was examined in a group of 12 visually normal subjects, using sinusoidal full-field flicker and harmonic analysis of the ERG waveforms. For all subjects, the ERG responses to 32-Hz flicker (a frequency commonly used clinically) were characterized by a harmonic component at the stimulus frequency and at higher harmonics that were integral multiples of the stimulus frequency, as expected. In addition, six of the subjects showed period doubling at 32 Hz, characterized by harmonic components at integer multiples of a frequency that was half the stimulus frequency (the subharmonic). However, the subharmonic itself did not exceed the noise level. These findings suggest that the subharmonic is generated prior to or at the site that produces the nonlinear higher harmonics of the ERG response, and that a subsequent band-pass filter attenuates this subharmonic. Examination of harmonic components of the subjects' ERG waveforms at other stimulus frequencies, as well as a cycle-by-cycle analysis of the ERG waveforms, suggested that individual differences in period doubling may be due to intersubject variation in the strength of the hypothesized feedback signal and/or the time constant of its decay. PMID- 16469191 TI - Gycine and GABA interact to regulate the nitric oxide/cGMP signaling pathway in the turtle retina. AB - Nitric oxide (NO) is a free radical that is important in retinal signal transduction and cyclic guanosine monophosphate (cGMP) is a critical downstream messenger of NO. The NO/cGMP signaling pathway has been shown to modulate neurotransmitter release and gap junction coupling in horizontal cells and amacrine cells, and increase the gain of the light response in photoreceptors. However, many of the mechanisms controlling the production of NO and cGMP remain unclear. Previous studies have shown activation of NO/cGMP production in response to stimulation with N-methyl-d-aspartate (NMDA) or nicotine, and the differential modulation of cGMP production by GABA(A) and GABA(C) receptors (GABA(A)Rs and GABA(C)Rs). This study used cGMP immunocytochemistry and NO imaging to investigate how the inhibitory GABAergic and glycinergic systems modulate the production of NO and cGMP. Our data show that blocking glycine receptors (GLYR) with strychnine (STRY) produced moderate increases in cGMP-like immunoreactivity (cGMP-LI) in select types of amacrine and bipolar cells, and strong increases in NO-induced fluorescence (NO-IF). TPMPA, a selective GABACR antagonist, greatly reduced the increases in cGMP-LI stimulated by STRY, but did not influence the increase in NO-IF stimulated by STRY. Bicuculline (BIC), a GABA(A)R antagonist, however, enhanced the increases in both the cGMP-LI and NO-IF stimulated by STRY. CNQX, a selective antagonist for alpha-Amino-3-hydroxy-5-methyl-4 isoxazoleproprionic acid hydrobromide/kainic acid (AMPA/KA) receptors, eliminated both the increases in cGMP-LI and NO-IF stimulated by STRY, while MK801, a selective antagonist for NMDA receptors, slightly increased the cGMP-LI and slightly decreased the NO-IF stimulated by STRY. Finally, double labeling of NO stimulated cGMP and either GLY or GABA indicated that cGMP predominantly colocalized with GLY. Taken together, these findings support the hypothesis that GLY and GABA interact in the regulation of the NO/cGMP signaling pathway, where GLY primarily inhibits NO production and GABA has a greater effect on cGMP production. Such interacting inhibitory pathways could shape the course of signal transduction of the NO/cGMP pathway under different physiological situations. PMID- 16469192 TI - Muscimol and baclofen differentially suppress retinotopic and nonretinotopic responses in visual cortex. AB - This study relates to local field potentials and single-unit responses in cat visual cortex elicited by contrast reversal of bar gratings that were presented in single, double, or multiple discrete patch (es) of the visual field. Concurrent stimulation of many patches by means of the pseudorandom, binary m sequence technique revealed interactions between their respective responses. An analysis identified two distinct components of local field potentials: a fast local component (FLC) and a slow distributed component (SDC). The FLC is thought to be a primarily postsynaptic response, as judged by its relatively short latency. It is directly generated by thalamocortical volleys following retinotopic stimulation of receptive fields of a small cluster of single cells, combined with responses to recurrent excitation and inhibition derived from the cells under study and immediately neighboring cells. In contrast, the SDC is thought to be an aggregate of dendritic potentials related to the long-range lateral connections (i.e. long-range coupling). We compared the suppressive effects of a GABA(A)-receptor agonist, muscimol, on the FLC and SDC with those of a GABA(B)-receptor agonist, baclofen, and found that muscimol more strongly suppressed the FLC than the SDC, and that the reverse was the case for baclofen. The differential suppression of the FLC and SDC found in the present study is consistent with the notion that intracortical electrical signals related to the FLC terminate on the somata and proximal/basal dendrites, while those related to the SDC terminate on distal dendrites. PMID- 16469194 TI - Cone visual pigments of aquatic mammals. AB - It has long been hypothesized that the visual systems of animals are evolutionarily adapted to their visual environment. The entrance many millions of years ago of mammals into the sea gave these new aquatic mammals completely novel visual surroundings with respect to light availability and predominant wavelengths. This study examines the cone opsins of marine mammals, hypothesizing, based on previous studies [Fasick et al. (1998) and Levenson & Dizon (2003)], that the deep-dwelling marine mammals would not have color vision because the pressure to maintain color vision in the dark monochromatic ocean environment has been relaxed. Short-wavelength-sensitive (SWS) and long wavelength-sensitive (LWS) cone opsin genes from two orders (Cetacea and Sirenia) and an additional suborder (Pinnipedia) of aquatic mammals were amplified from genomic DNA (for SWS) and cDNA (for LWS) by PCR, cloned, and sequenced. All animals studied from the order Cetacea have SWS pseudogenes, whereas a representative from the order Sirenia has an intact SWS gene, for which the corresponding mRNA was found in the retina. One of the pinnipeds studied (harp seal) has an SWS pseudogene, while another species (harbor seal) appeared to have an intact SWS gene. However, no SWS cone opsin mRNA was found in the harbor seal retina, suggesting a promoter or splice site mutation preventing transcription of the gene. The LWS opsins from the different species were expressed in mammalian cells and reconstituted with the 11-cis-retinal chromophore in order to determine maximal absorption wavelengths (lambda(max)) for each. The deeper dwelling Cetacean species had blue shifted lambda(max) values compared to shallower dwelling aquatic species. Taken together, these findings support the hypothesis that in the monochromatic oceanic habitat, the pressure to maintain color vision has been relaxed and mutations are retained in the SWS genes, resulting in pseudogenes. Additionally, LWS opsins are retained in the retina and, in deeper dwelling animals, are blue shifted in lambda(max). PMID- 16469193 TI - Homotypic constraints dominate positioning of on- and off-center beta retinal ganglion cells. AB - Beta retinal ganglion cells (RGCs) of the cat are classified as either on-center or off-center, according to their response to light. The cell bodies of these on- and off-center RGCs are spatially distributed into regular patterns, known as retinal mosaics. In this paper, we investigate the nature of spatial dependencies between the positioning of on- and off-center RGCs by analysing maps of RGCs and simulating these patterns. We introduce principled approaches to parameter estimation, along with likelihood-based techniques to evaluate different hypotheses. Spatial constraints between cells within-type and between-type are assumed to be controlled by two univariate interaction functions and one bivariate interaction function. By making different assumptions on the shape of the bivariate interaction function, we can compare the hypothesis of statistical independence against the alternative hypothesis of functional independence, where interactions between type are limited to preventing somal overlap. Our findings suggest that the mosaics of on- and off-center beta RGCs are likely to be generated assuming functional independence between the two types. By contrast, allowing a more general form of bivariate interaction function did not improve the likelihood of generating the observed maps. On- and off-center beta RGCs are therefore likely to be positioned subject only to homotypic constraints and the physical constraint that no two somas of opposite type can occupy the same position. PMID- 16469195 TI - Association amacrine cells of Ramon y Cajal: rediscovery and reinterpretation. AB - In 1895, by means of the Golgi method, Santiago Ramon y Cajal discovered a cell having a unique morphology in the avian retina. This cell had its cell body in the amacrine cell level of the inner nuclear layer, only a few rudimentary dendrites at the outermost level of the inner plexiform layer (IPL), and a long axon coursing horizontally and terminating in the IPL. Despite having defined amacrine cells as cells without axons, Cajal named this cell type "association amacrine cell" (AAC). This discovery was not confirmed by other investigators for nearly a century. Very recently, however, isthmo-optic target cells (IOTCs), which receive the terminals of centrifugal fibers emanating from the isthmo-optic nucleus, have been identified as one type of AAC. As summarized and discussed in this review, the morphology of the AACs as described by Cajal has been completely confirmed. However, since these cells appear to be classical polarized, monoaxonal neurons and lack the dendritic interactions that are typical of amacrine cells, they should be regarded as a distinct type of retinal interneuron and not as amacrine cells. PMID- 16469196 TI - Cone and rod inputs to murine retinal ganglion cells: evidence of cone opsin specific channels. AB - To identify ultraviolet (UV) and middle- (M) wavelength-sensitive cone and rod signals in murine retinal ganglion cells, single ganglion cell responses were studied in anesthetized, light-adapted C57/BL6 mice with tungsten microelectrodes driven through the sclera and vitreous to the neural retina. One hundred fifty four ganglion cells were examined in 43 retinas of 34 mice. The retina was stimulated with diffuse flashes and/or pulses of ultraviolet (360 nm) or green (520 nm) light in the presence and absence of a strong steady orange adapting light. Twelve ganglion cells were studied in the dark-adapted retina in order to identify the signals of rods. Three functionally different types of ganglion cells were found: (1) phasic responding cells (31%) with no spontaneous activity and large impulse amplitudes; (2) tonic responding cells (60%) with irregular, low frequency (5-10 Hz) spontaneous activity and smaller impulse amplitudes; and (3) metronome-like cells (9%) with regular, relatively high-frequency (20-40 Hz) spontaneous activity. A few cells (1%) had habituating responses. Every cell encountered was affected by diffuse stimulation. The more common two types were excited at either the ON or OFF or at both the ON and OFF phases of stimulation. Type III cells had weaker responses, sometimes only inhibited by turning off a light. In the light-adapted state, most cells received signals of the same polarity from UV- and M-cones but UV-cone inputs were usually more dominant, especially in ventral retina. A fraction of cells received signals from only UV- (18%) or only M- (3%) cones. In rare cases (2%) these cone inputs had an opposite polarity on the same cell. In the dark-adapted state, all cells were at least four or five logarithmic units more sensitive and more to green than ultraviolet light. The results indicate that co-expression of both UV-and M-cone opsins cannot be ubiquitous in murine retina. Some cones, especially UV cones, exist without the presence of any functional M-cone opsin. This must be the case to explain the presence of ganglion cells that receive inputs only from UV-cones and others that receive inputs of opposite polarity from UV- and M-cones. The results support the hypothesis that murine retina has the physiological capacity to relay signals to the brain that allow the sensing of chromatic contrast and color vision. PMID- 16469198 TI - Structural brain correlates of IQ changes in bipolar disorder. AB - BACKGROUND: There is increasing evidence that cognitive deficits are present in bipolar disorder (BP), but their neural correlates have not been fully explored. The aim of this study is to correlate structural brain abnormalities with cognitive performance in BP and to explore differences between clinical subtypes. METHOD: Thirty-six BP patients (13 men, 23 women) with a mean age of 39 years (range 21-63 years) underwent neuropsychological testing and imaging. Twenty-five patients had bipolar disorder I (BP I) and 11 had bipolar disorder II (BP II). Patients with co-morbid psychiatric diagnosis, drug and alcohol abuse or systemic illness were excluded. Correlations between cognitive performance and structural brain changes were explored using high-resolution anatomical imaging and magnetization transfer imaging (MTI). RESULTS: In the whole BP group the difference between estimated pre-morbid IQ and current IQ was significantly correlated with left-sided reduction of the magnetization transfer ratio (MTR) in the superior temporal gyrus, uncus and para-hippocampal gyrus. In BP II patients the areas where these correlations were significant extended to the right superior and middle temporal gyri, cingulate gyrus, pre-cuneus and adjacent frontal and parietal white matter. The volume of superior temporal white matter was also correlated with IQ difference in this subgroup. CONCLUSIONS: The study highlights the association between fronto-temporal abnormalities and decline in IQ in BP. The more extensive abnormalities present in BP II patients suggest that persistent depression, rather than mania, may be a key pathophysiological factor or that BP II represents a clinical phenotype with a higher risk of developing cognitive abnormalities. PMID- 16469199 TI - Expression of two cell wall proteins during the intracellular development of Encephalitozoon cuniculi: an immunocytochemical and in situ hybridization study with ultrathin frozen sections. AB - The microsporidian Encephalitozoon cuniculi is an obligate intracellular parasite that develops asynchronously inside parasitophorous vacuoles. Spore differentiation involves the construction of a cell wall commonly divided into an outer layer (exospore) and a thicker, chitin-rich inner layer (endospore). The developmental patterns of protein deposition and mRNA expression for 2 different spore wall proteins were studied using immunocytochemical and in situ hybridization procedures with ultrathin frozen sections. The onset of deposition of an exospore-destined protein (SWP1) correlated with the formation of lamellar protuberances during meront-to-sporont conversion. No evidence for a release of SWP1 towards the parasitophorous vacuole lumen was obtained. An endospore destined protein (EnP1) was detected early on the plasma membrane of meronts prior to extensive accumulation within the chitin-rich layer of sporoblasts. swp1 mRNA was preferentially synthesized in early sporogony while enp1 mRNA was transcribed during merogony and a large part of sporogony. The level of both mRNAs was reduced in mature spores. Considering the availability of the E. cuniculi genome sequence, the application of nucleic and/or protein probes to cryosections should facilitate the screening of various genes for stage-specific expression during microsporidian development. PMID- 16469201 TI - Morphological and molecular characterization of a new microsporidian (Protozoa: Microsporidia) isolated from Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae). AB - A microsporidium was isolated from larvae of Spodoptera litura (Fabricius) collected from Tamil Nadu, India. This microsporidian species is monomorphic, disporous and develops in direct contact with the cytoplasm of the host cell. The nuclear configuration of merogonic and sporogonic stages was diplokaryotic. The merogonic proliferative stage was unusual that normal development with 1, 2 and 4 binucleated forms were common, while large multinucleate meronts containing 8 and 12 small compact horseshoe-like diplokaryotic nuclei were also observed. The fresh spores were typically ovocylindrical in shape, with a mean size of 3.91 x 1.91 microm and the polar filament length was approximately 90 microm. Infection was systemic with mature spores produced in the midgut, nervous tissue, muscles, labial glands, gonads, tracheae, epidermis, Malpighian tubules and, most extensively, fat body tissues. The new isolate was highly pathogenic to S. litura larvae. Host specificity tests performed on 37 non-target hosts of 5 different insect orders revealed that the new isolate is pathogenic only to lepidopteran insects. We sequenced the 16S small subunit rRNA (SSU rRNA) gene of the isolate and compared it with 72 non-redundant microsporidian sequences from the GenBank. Based on the light microscopic studies and phylogenetic analyses, the new isolate is assigned to the genus Nosema. Significant differences in the SSU rRNA sequence were identified when compared with the type species Nosema bombycis and other closely related species viz., Nosema spodopterae. Structural differences were also observed in the 16S SSU rRNA between the new isolate and the two above mentioned microsporidian pathogens. We conclude that the microsporidian isolate reported here is distinctly different from the other known species and is likely to be a new species. PMID- 16469200 TI - Co-infection with Onchocerca volvulus and Loa loa microfilariae in central Cameroon: are these two species interacting? AB - Ivermectin treatment may induce severe adverse reactions in some individuals heavily infected with Loa loa. This hampers the implementation of mass ivermectin treatment against onchocerciasis in areas where Onchocerca volvulus and L. loa are co-endemic. In order to identify factors, including co-infections, which may explain the presence of high L. loa microfilaraemia in some individuals, we analysed data collected in 19 villages of central Cameroon. Two standardized skin snips and 30 mul of blood were obtained from each of 3190 participants and the microfilarial (mf) loads of both O. volvulus and L. loa were quantified. The data were analysed using multivariate hierarchical models. Individual-level variables were: age, sex, mf presence, and mf load; village-related variables included the endemicity levels for each infection. The two species show a certain degree of ecological separation in the study area. However, for a given individual host, the presence of microfilariae of one species was positively associated with the presence of microfilariae of the other (OR=1.79, 95% CI [1.43-2.24]). Among individuals harbouring Loa microfilariae, there was a slight positive relationship between the L. loa and O. volvulus mf loads which corresponded to an 11% increase in L. loa mf load per 100 O. volvulus microfilariae. Co-infection with O. volvulus is not sufficient to explain the very high L. loa mf loads harboured by some individuals. PMID- 16469202 TI - An independent non-linear latitudinal cline for the sn-glycerol-3-phosphate (alpha- Gpdh ) polymorphism of Drosophila melanogaster from eastern Australia. AB - Latitudinal variation of the polymorphic sn-glycerol-3-phosphate (alpha-Gpdh) locus in Drosophila melanogaster has been characterized on several continents; however, apparent clinal patterns are potentially confounded by linkage with an inversion, close associations with other genetic markers that vary clinally, and a tandem alpha-Gpdh pseudogene. Here we compare clinal patterns in alpha-Gpdh with those of other linked markers by testing field flies from eastern Australian locations collected in two separate years. The alpha-Gpdh variation exhibited a consistent non-linear cline reflecting an increase in the alpha-GpdhF allele at extreme latitudes. This pattern was not influenced by the In(2L)t inversion wherein this locus is located, nor was it influenced by the presence of the alpha Gpdh pseudogene, whose presence was ubiquitous and highly variable among populations. The alpha-Gpdh pattern was also independent of a cline in allozyme frequencies at the alcohol dehydrogenase (Adh) locus, and two length polymorphisms in the Adh gene. These results suggest clinal selection at the alpha-Gpdh locus that is partially or wholly unrelated to linear climatic gradients along the eastern coast of Australia. PMID- 16469203 TI - An effective correlation dimension and burst suppression ratio of the EEG in rat. Correlation with sevoflurane induced anaesthetic depth. AB - BACKGROUND AND OBJECTIVE: Anaesthesiologists need parameters that measure the depth of anaesthesia. In the context of this need, the present study investigated in rats how two variables from the electroencephalogram, the burst suppression ratio and effective correlation dimension correlated with a measure of anaesthetic depth as measured in the strength of a noxious withdrawal reflex. METHODS: Eight rats were exposed to different inspiratory concentrations of sevoflurane, each rat in two separate experiments. In the first experiment, spontaneously breathing animals could move freely and no painful stimuli were applied. In the second experiment, in mechanically ventilated restrained anaesthetized rats, the withdrawal reflex was measured every 80 s. In both experiments the electroencephalogram was continuously recorded. The concentration in the effector compartment was estimated using a first order two compartment model. Correlation dimension was computed following the Grassberger/Procaccia/Takens approach with optimized parameter settings to achieve maximum sensitivity to anaesthetic drug effects and enable real-time computation. The Hill, equation was fitted to the data, describing the effect as a function of sevoflurane concentration. RESULTS: Good correlations of Depth of Anaesthesia with correlation dimension as well as burst suppression ratio were established in both types of experiments. Arousal by noxious stimuli decreased burst suppression ratio and increased correlation dimension. The effective sevoflurane concentration associated with 50% of the maximum effect (C50) was higher in experiment II (stimulation) than in experiment I (no stimulation): i.e. for correlation dimension 2.18% vs. 0.60% and for burst suppression ratio 3.07% vs. 1.73%. The slope factors were: gammaCD = 4.15 vs. gammaCD = 1.73 and gammaBSR = 5.2 vs. gammaBSR = 5.4. Correlation dimension and burst suppression ratio both correlated with the strength of the withdrawal reflex with correlation coefficients of 0.46 and 0.66 respectively (P < 0.001). CONCLUSIONS: Both correlation dimension and burst suppression ratio are related to anaesthetic depth and are affected by noxious stimuli. The relationship between anaesthetic depth and burst suppression ratio is confirmed and the potential of correlation dimension is demonstrated. PMID- 16469204 TI - Predictors of mortality in ARDS patients referred to a tertiary care centre: a pilot study. AB - BACKGROUND AND OBJECTIVE: In order to identify parameters predicting intensive care unit mortality in patients transferred to a specialized tertiary centre because of progressive acute respiratory distress syndrome, an observational pilot study was carried out involving 94 patients. METHODS AND RESULTS: Forty-one patients (43.6%) died. Survival was defined as intensive care unit discharge. Survivors were younger (32.0 +/- 11.8 vs. 39.1 +/- 12.4 yr, P = 0.008), at admission they had a lower acute physiology and chronic health evaluation (APACHE) II score (21.7 +/- 5.4 vs. 25.4 +/- 5.2, P = 0.0009), higher PaO2/FiO2 (122 +/- 79 vs. 79 +/- 42 mmHg, P = 0.002), lower positive end-expiratory pressure (10.6 +/- 3.1 vs. 12.5 +/- 3.7 cmH2O, P = 0.02) and a lower Murray score (2.8 +/- 0.63 vs. 3.0 +/- 0.62, P = 0.04). No differences were observed for tidal volumes and peak inspiratory pressures. Days of hospitalization and mechanical ventilation prior to transferral were not related to survival. Multivariate analysis of variables assessed on admission detected only differences for age (P = 0.014) and APACHE II (P = 0.005). Odds ratio was 1.06 (95% confidence interval (CI): 1.013-1.119) for age and 1.21 (CI: 1.059-1.381) for APACHE II. Multivariate analysis of changes in respiratory parameters, APACHE II and Murray score during the first 3 days after transferral revealed a significant difference only for positive end-expiratory pressure (P < 0.008). Corresponding odds ratio was 2.40 (CI: 1.25-4.58) for an increase of 1 cmH2O/24 h. CONCLUSION: Age-related mortality in this small, but highly selected group of patients with established ARDS increased early in life even in a population with an overall mean age of 35.1 yr. APACHE II was the only clinical predictor for mortality on admission. The need for a substantial increase in positive end-expiratory pressure after transferral markedly reduced the chance to survive. PMID- 16469205 TI - Stress and burnout among Portuguese anaesthesiologists. AB - BACKGROUND AND OBJECTIVE: Anaesthesiology is considered a stressful occupation. Our purpose was to assess stress and burnout among Portuguese anaesthesiologists. METHODS: A cross-sectional survey based on an anonymous questionnaire was sent to all Portuguese anaesthesiologists registered by the Portuguese Medical Association. Data on patient characteristics and professional variables, perceived stress (Perceived Stress Scale), the subscale of work satisfaction of the Pressure Management Indicator scale, burnout (Maslach Burnout Inventory) and main stress factors were collected. The proportion of participation was 31.8%. RESULTS: The average value of perceived stress scale was 24.0 (range: 0-56). Among the 263 anaesthesiologists, 57.9% experienced emotional exhaustion, 44.8% lack of personal accomplishment and 90.9% depersonalisation. Lack of personal accomplishment increased with number of children among women but not among men; depersonalisation was more frequent among anaesthesiologists working in community hospitals and anaesthesiologists with leadership functions experienced less professional stress. CONCLUSION: There are stress conditions and burnout amongst Portuguese anaesthesiologists. The prevalence of depersonalisation was extremely high in the studied sample. Emotional exhaustion is partially explained by high perceived-stress and low satisfaction with organisation in the job according to Pressure Management Indicator scale. PMID- 16469206 TI - The impact of intraoperative propofol administration in the prevention of postoperative pruritus induced by epidural morphine. AB - BACKGROUND AND OBJECTIVE: We examined the efficacy of intraoperative propofol administration to prevent pruritus induced by epidural morphine. METHODS: Seventy patients ASA I-II undergoing combined epidural and general anaesthesia for hysterectomy were randomly assigned to two groups, Group P where anaesthesia was induced with propofol and fentanyl and maintained with propofol-nitrous oxide and Group S in which anaesthesia was induced with thiopental and fentanyl and maintained with sevoflurane-nitrous oxide. All patients received a ropivacaine epidural bolus with 3 mg morphine 1 h before the end of surgery. The incidence and severity of pruritus were evaluated every 4 h for the first 12 h postoperatively by blinded observers. RESULTS: The total incidence of pruritus was significantly higher (P = 0.024) in Group S (65.6%) compared to Group P (29%) between 4 and 8 h postoperatively. There were also significantly more patients (P = 0.03) reporting severe pruritus in Group S (22%) compared to Group P (0). CONCLUSION: Propofol-based general anaesthesia compared to thiopental-sevoflurane based anaesthesia reduces the incidence and severity of pruritus induced by a single injection of 3 mg epidural morphine with ropivacaine. PMID- 16469207 TI - Effect of sevoflurane preconditioning on ischaemia/reperfusion injury in the rat kidney in vivo. AB - BACKGROUND AND OBJECTIVE: Whereas the protective effect of anaesthetic and ischaemic preconditioning has been described for several organs, it is uncertain whether this mechanism is also effective in the kidney. We compared the effect of preconditioning with sevoflurane and preconditioning with short episodes of ischaemia on renal ischaemia/reperfusion injury in the rat in vivo. METHODS: Fourteen days after right-sided nephrectomy, anaesthetized male Wistar rats were randomly assigned to a sham-operated group (no arterial occlusion, n = 5) or underwent 45 min of left renal artery occlusion (control group, n = 9) followed by 3 days of reperfusion. Two further experimental groups of animals were preconditioned prior to ischaemia either by administering 1 MAC sevoflurane for 15 min followed by 10 min of washout (sevoflurane group, n = 10) or by subjecting the animals to three short episodes of renal ischaemia (ischaemia-preconditioned group, n = 8). Blood creatinine was measured during reperfusion and morphological damage was assessed by histological examination. RESULTS: Baseline creatinine values were similar in all four groups (0.7 +/- 0.2 mg dL-1; mean +/- SD) and remained unchanged in the sham-operated animals after 3 days (0.8 +/- 0.2 mg dL 1). Creatinine levels increased in the ischaemic preconditioning group (3.3 +/- 1.2 mg dL-1) and sevoflurane preconditioning group (4.0 +/- 1.1 mg dL-1) compared to the control group (1.6 +/- 0.6 mg dL-1). Morphological damage was less severe in the control group, i.e. in animals without preconditioning, than in both preconditioning groups. CONCLUSION: Neither sevoflurane nor ischaemic preconditioning preserves renal function or attenuates cell damage in the rat in vivo. PMID- 16469208 TI - The effect of a remifentanil bolus on the bispectral index of the EEG (BIS) in anaesthetized patients independently from intubation and surgical stimuli. AB - BACKGROUND AND OBJECTIVE: Remifentanil boluses are used in different clinical situations and the effects on bispectral index monitoring are unclear. We analysed the effect of a remifentanil bolus on the bispectral index of the electroencephalogram (bispectral index) under total intravenous anaesthesia with propofol and remifentanil. METHODS: ASA I-III patients were included in this study. All patients received a 2 microg k g-1 remifentanil bolus in a period free from stimuli. Bispectral index and haemodynamic data were collected from an A 2000XP bispectral index monitor (every second) and an AS/3 Datex monitor (every 5 s). Bispectral index data were analysed using the area under the curve. Mean arterial pressure and heart rate were averaged at each 30-s period and analysed using analysis of variance. RESULTS: A total of 240 bispectral index values were obtained per patient. The area under the curve between 90 and 120 s after the bolus was significantly lower than the basal area under the curve (average of all areas before the bolus, P < 0.05). Mean arterial pressure and heart rate were significantly reduced from 96.4 +/- 19.9 mmHg at the time of the bolus to 74.2 +/ 16.6 mmHg 120 s after, and from 70 +/- 16.4 bpm at the time of the bolus to 61 +/- 13.6 bpm after (P < 0.001), respectively. CONCLUSIONS: There was a significant reduction in the areas under the curve between 90-120 s following the bolus. Heart rate and blood pressure also showed significant reductions. Thus, remifentanil bolus given under total intravenous anaesthesia with propofol and remifentanil decreases bispectral index, an effect independent of intubation and surgical stimuli. PMID- 16469209 TI - Economic impact of bariatrics on a general surgery practice. AB - Bariatric surgery is currently a rapidly growing subsection of general surgery, with exponential expansion over the past decade. Many residency programs lacked sufficient experience in bariatrics, necessitating established surgeons to consider re-training and re-vamping of their practice to enter the field. The addition of bariatric surgery to a general surgery practice can present economic consequences, which are both positive and negative. Positive consequences include a potential new revenue source with a large population base. Negative consequences include increased employees, required paper-work and office resources, increased malpractice premiums, difficulties with appropriate reimbursement, and limitations on access to appointment time for non-bariatric cases. This paper reviews the potential economic impact of bariatric surgery on a general surgery practice and possible alternatives to manage these efficiently. PMID- 16469210 TI - Laparoscopic surgery for morbid obesity: 1,001 consecutive bariatric operations performed at The Bariatric Institute, Cleveland Clinic Florida. AB - BACKGROUND: Morbid obesity is an epidemic in America. This series evaluates the safety and efficacy in the first 1,001 laparoscopic bariatric operations performed at The Bariatric Institute, Cleveland Clinic Florida. METHODS: A retrospective review was conducted examining all patients undergoing a primary bariatric procedure (either laparoscopic gastric bypass or laparoscopic gastric banding) from July 2000 to December 2003. RESULTS: 2 surgeons performed 1,001 laparoscopic bariatric operations. Average age was 47 (19-75) years, average BMI was 55.6 (35-97) kg/m2, and average ASA class was III. Excess weight loss was 51% at 6 months, 73.4% at 1 year for the gastric bypass group and 54% at 1 year for the laparoscopic banding group. The overall complication rate was 31.8% (12.4% major and 19.4% minor) in the gastric bypass group and 13% in the laparoscopic banding group. There was no postoperative mortality. CONCLUSION: Laparoscopic bariatric surgery is feasible and safe for weight loss. Results obtained have been comparable to those reported for the open approach for weight loss, with a similar major morbidity rate and an improved mortality rate. PMID- 16469211 TI - Clinical evaluation of fibrin glue in the prevention of anastomotic leak and internal hernia after laparoscopic gastric bypass: preliminary results of a prospective, randomized multicenter trial. AB - BACKGROUND: Gastro-jejunal anastomotic leak and internal hernia can be life threatening complications of laparoscopic Roux-en-Y gastric bypass (LRYGBP), ranging from 0.1-4.3% and from 0.8-4.5% respectively. The safety and efficacy of a fibrin glue (Tissucol) was assessed when placed around the anastomoses and over the mesenteric openings for prevention of anastomotic leaks and internal hernias after LRYGBP. METHODS: A prospective, randomized, multicenter, clinical trial commenced in January 2004. Patients with BMI 40-59 kg/m2, aged 21-60 years, undergoing LRYGBP, were randomized into: 1) study group (fibrin glue applied on the gastro-jejunal and jejuno-jejunal anastomoses and the mesenteric openings); 2) control group (no fibrin glue, but suture of the mesenteric openings). 322 patients, 161 for each arm, will be enrolled for an estimated period of 24 months. Sex, age, operative time, time to postoperative oral diet and hospital stay, early and late complications rates are evaluated. An interim evaluation was conducted after 15 months. RESULTS: To April 2005, 204 patients were randomized: 111 in the control group (mean age 39.0+/-11.6 years, BMI 46.4 +/- 8.2) and 93 in the fibrin glue group (mean age 42.9+/-11.7 years, BMI 46.9+/-6.4). There was no mortality or conversion in both groups; no differences in operative time and postoperative hospital stay were recorded. Time to postoperative oral diet was shorter for the fibrin glue group (P = 0.0044). Neither leaks nor internal hernias have occurred in the fibrin glue group. The incidence of leaks (2 cases, 1.8%) and the overall reoperation rate were higher in the control group (P=0.0165). CONCLUSION: The preliminary results suggest that Tissucol application has no adverse effects, is not time-consuming, and may be effective in preventing leaks and internal hernias in morbidly obese patients undergoing LRYGBP. PMID- 16469212 TI - Prevention of pouch dilatation after laparoscopic adjustable gastric banding. AB - BACKGROUND: The major long-term complication of laparoscopic adjustable gastric banding (LAGB) is dilatation of the gastric pouch, that is reported with a frequency ranging from 1 to 25%, and often requires removal of the band. In addition to the usual recommendations of bariatric surgery centers and dietetic advice to prevent this complication, over the last 4 years we introduced a technical modification of the procedure. METHODS: From Nov 1993 to Dec 2004, 684 morbidly obese patients underwent adjustable gastric banding, 83 patients by open surgery and 601 patients by laparoscopy. The first 323 patients (group A) were operated by the perigastric approach, and 57 patients (group B) were operated by the pars flaccida approach. Since Dec 2000, 304 patients (group C) were operated with a modified pars flaccida technique, which consisted in suturing the gastric lesser curvature below the band with one or two stitches to the right phrenic crus to secure the band in place. RESULTS: In group A, the most important late complication was irreversible dilatation of the gastric pouch, which occurred in 35 patients (10.8%), and required removal of the band in 30 cases and replacement in 5. In group B, there were 3 pouch dilatations (5.2%). In group C, only 4 dilatations occurred (1.31%), which required 3 band removals and 1 band replacement. CONCLUSION: Dilatation of the gastric pouch appears to be dramatically reduced by our minor technical modification of band placement. PMID- 16469213 TI - Revision of failed laparoscopic adjustable gastric banding to Roux-en-Y gastric bypass. AB - BACKGROUND: The most common bariatric surgical operation in Europe, laparoscopic adjustable gastric banding (LAGB), is reported to have a high incidence of long term complications. Also, insufficient weight loss is reported. We investigated whether revision to Roux-en-Y gastric bypass (RYGBP) is a safe and effective therapy for failed LAGB and for further weight loss. METHODS: From Jan 1999 to May 2004, 613 patients underwent LAGB. Of these, 47 underwent later revisional Roux-en-Y gastric bypass (RYGBP). Using a prospectively collected database, we analyzed these revisions. All procedures were done by two surgeons with extensive experience in bariatric surgery. RESULTS: All patients were treated with laparoscopic (n=26) or open (n=21) RYGBP after failed LAGB. Total follow-up after LAGB was 5.5+/-2.0 years. For the RYGBP, mean operating time was 161+/-53 minutes, estimated blood loss was 219+/-329 ml, and hospital stay was 6.7+/-4.5 days. There has been no mortality. Early complications occurred in 17%. There was only one late complication (2%)--a ventral hernia. The mean BMI prior to any form of bariatric surgery was 49.2+/-9.3 kg/m2, and decreased to 45.8+/-8.9 kg/m2 after LAGB and was again reduced to 37.7+/-8.7 kg/m2 after RYGBP within our follow-up period. CONCLUSION: Conversion of LAGB to RYGBP is effective to treat complications of LAGB and to further reduce the weight to healthier levels in morbidly obese patients. PMID- 16469214 TI - Value of preoperative upper endoscopy in patients undergoing laparoscopic gastric bypass. AB - BACKGROUND: Preoperative evaluation of patients undergoing laparoscopic Roux-en-Y gastric bypass (LRYGBP) has included esophagogastroduodenoscopy (EGD) with little data to substantiate its use. METHODS: A retrospective analysis was conducted of patients from Feb 04 to Mar 05 who underwent preoperative EGD and subsequently LRYGBP. RESULTS: 169 patients underwent EGD prior to surgery. Their mean age was 41.1 years (range 14-66), mean BMI 49.7 (range 35-78), and 82% were females. There were no complications from EGD. Significant findings in patients at EGD included gastric ulceration in 3 (2%), duodenal ulcer in 1 (0.7%), Barrett's esophagus in 2 (1.3%), and a GI stromal tumor (GIST) in 1 (0.7%). EGD revealed hiatal hernias in 56 (35.2%), esophagitis in 28 (17%), Schatzki's ring in 5 (3%), gastritis in 43 (27%), gastric polyps in 8 (5%), and duodenitis in 9 (6%). 53 patients (33.3%) had a negative EGD. Ulcer and severe gastritis, esophagitis, and duodenitis diagnosed preoperatively were treated medically before surgery. 9 hiatal hernias were repaired intraoperatively. The patient with the GIST underwent laparoscopic near-total gastrectomy and gastric bypass, while 1 patient with an antral polyp underwent laparoscopic partial gastrectomy in addition to the LRYGBP. CONCLUSION: EGD is essential for diagnosis of GI diseases including tumors, ulcers, and hiatal hernias that alter the medical and surgical management of patients undergoing gastric bypass. PMID- 16469215 TI - Intragastric balloon for preoperative weight reduction in candidates for laparoscopic gastric bypass with massive obesity. AB - BACKGROUND: Laparoscopic bariatric operations can be technically challenging in massively obese patients. The potential of the intragastric balloon for preoperative weight reduction was evaluated in candidates for laparoscopic Roux en-Y gastric bypass (LRYGBP) with super obesity. METHODS: From January 2004 to March 2005, 10 patients with super obesity who were potential candidates for LRYGBP were included in a prospective observational study. An intragastric balloon was placed endoscopically under general anesthesia. Patients were then followed by a multidisciplinary team until removal of the balloon after 6 months. Weight, BMI, and percent excess weight lost (%EWL) were monitored after 1, 3 and 6 months. RESULTS: The 10 patients were all female with mean +/- SD age of 33+/ 11 years (range 17-51). Initial weight was 175+/-25 kg (range 135-223) and initial BMI was 64+/-7 kg/m2 (range 59-78). After completion of 1, 3 and 6 months, weight, BMI, and %EWL reached respectively: 166+/-27 kg*, 165+/-27 kg*, and 169+/-26 kg*; 61.1+/-7.6 kg/m2*, 60.8+/-8.4 kg/m2*, and 61+/-8.2 kg/m2*; 9+/ 5%, 10+/-7%, 7+/-6%. (*=P<0.01 vs preop). CONCLUSION: In potential candidates for LRYGBP with super obesity, preoperative placement of an intragastric balloon can reduce the excess weight by 10% within 3 months. Extending this period failed to improve these results further. PMID- 16469216 TI - A prospective comparison of vertical banded gastroplasty and Roux-en-Y gastric bypass in a non-superobese population. AB - BACKGROUND: In the non-superobese population, consensus is currently unavailable in bariatric surgery. We report the results of a prospective comparison of vertical banded gastroplasty (VBG) and Roux-en-Y gastric bypass (RYGBP) in a non superobese population. METHODS: From 1994 to 2000, 179 patients with clinically severe obesity underwent various surgical procedures in our department. During this time a prospective study was undertaken in order to compare VBG with RYGBP in morbidly obese patients with a BMI <50 kg/m2. Based on specific criteria including eating behavior, 68 patients were selected to undergo RYGBP and 35 VBG. All patients have undergone complete follow-up evaluation at 1, 3, 6, and 12 months postoperatively and every year thereafter. RESULTS: All patients have now completed their 5th postoperative year. Mean follow-up period to date is 96.5+/ 12.2 months for VBG and 67.6+/-11.3 months for RYGBP. 3 patients (8.6%) in the VBG group and 9 patients (13.2%) in the RYGBP group are lost to follow-up. Mean excess weight loss (EWL) was always better in the RYGBP group (P=0.0013). The percentage of failure, defined as EWL <25%, was not significantly different between the two procedures. No statistically significant differences were observed between the 2 groups in the total number of non-metabolic complications, and the only statistically significant difference observed in metabolic complications was vitamin B12 deficiency after RYGBP. Frequency of vomiting was significantly less and quality of eating significantly better in RYGBP than in VBG patients. CONCLUSION: This prospective long-term study, with nearly complete follow-up, suggests that in the non-superobese population, preoperative eating habits may play a role in choosing the most appropriate bariatric operation for each patient. Although RYGBP is associated with better mean weight loss outcomes, the percentage of patients who achieved and maintained > or = 50% EWL after VBG in this pre-selected patient population was not significantly different. Each type of operation has advantages and disadvantages, and, if properly chosen, a purely restrictive procedure can be successful for some patients. Therefore, it can be said that the decision regarding which bariatric procedure to perform in non-superobese patients must be based on in-depth preoperative evaluation as well as the patients' own preferences and outcome expectations. PMID- 16469217 TI - Ethnic differences in obesity and surgical weight loss between African-American and Caucasian females. AB - BACKGROUND: In the general population, African-American females are more obese and resistant to weight loss than Caucasian women. In the present study, we examined the severity of obesity among morbidly obese African-American and Caucasian females, studied the effectiveness of Roux-en-Y gastric bypass (RYGBP), and sought to identify factors contributing to obesity and weight loss. METHODS: The study population included 153 morbidly obese females randomly selected from our general bariatric patient population. Anthropometric measurements consisted of body weight, body mass index (BMI), excess weight, and waist, hip, thigh, and neck circumferences. Factors that may contribute to obesity included age, age of obesity onset, number of childbirths, calorie intake, diet composition, and degree of psychological distress. The effects of RYBGP were studied in weight matched groups of African-American and Caucasian females (n=37 per group) at weight loss nadir, i.e. 12 to 18 months after surgery. RESULTS: We found that morbid obesity is more severe among African-American than Caucasian females. The greater degree of obesity of African-American, as compared to Caucasian, females is not due to ethnic differences in calorie intake, diet composition, age or age of obesity onset, number of childbirths, and psychological distress. RYGBP is less effective in reducing body fat and, consequently, excess body weight of the African-American than the Caucasian females, suggesting possible ethnic differences in fat metabolism. CONCLUSION: African-American females with morbid obesity have greater adiposity than do Caucasian women and lose significantly less body fat after RYGBP. PMID- 16469218 TI - Does gastric dilatation limit the success of sleeve gastrectomy as a sole operation for morbid obesity? AB - BACKGROUND: Sleeve gastrectomy as the sole bariatric operation has been reported for high-risk super-obese patients or as first-step followed by Roux-en-Y gastric bypass (RYGBP) or duodenal switch (DS) in super-super obese patients. The efficacy of laparoscopic sleeve gastrectomy (LSG) for morbidly obese patients with a BMI of <50 kg/m2 and the incidence of gastric dilatation following LSG have not yet been investigated. METHODS: 23 patients (15 morbidly obese, 8 super obese) were studied prospectively for weight loss following LSG. The incidence of sleeve dilatation was assessed by upper GI contrast studies in patients with a follow-up of >12 months. RESULTS: Patients who underwent LSG achieved a mean excess weight loss (EWL) at 6 and 12 months postoperatively of 46% and 56%, respectively. No significant differences were observed in %EWL comparing obese and super-obese patients. At a mean follow-up of 20 months, dilatation of the gastric sleeve was found in 1 patient and weight regain after initial successful weight loss in 3 of the 23 patients. CONCLUSION: LSG has been highly effective for weight reduction for morbid obesity even as the sole bariatric operation. Gastric dilatation was found in only 1 patient in this short-term follow-up. Weight regain following LSG may require conversion to RYGBP or DS. Follow-up will be necessary to evaluate long-term results. PMID- 16469219 TI - Evaluation of gastric greater curvature invagination for weight loss in rats. AB - BACKGROUND: Many bariatric endocopic or surgical procedures performed today reduce gastric capacity and/or induce an early sensation of satiety, alone or in combination with a distal enteric intervention. A form of prosthetic gastric wrap was used in the past for treating obesity with a high rate of reintervention. Nissen gastric fundoplication used in the treatment of gastroesophageal reflux disease induces a small but significant weight loss. We report the effect of gastric greater curvature invagination on weight in rats. METHODS: 30 rats were randomly divided into 3 groups. 10 rats in the first group (sham) were anesthesized and weighed. The rats from the second group (lap) were in addition submitted to a laparotomy plus visceral manipulation. In the third group (inv), invagination of the greater curvature of the stomach was added. All animals were weighed on the 7th and 21st days. They were then autopsied on the 21st day. RESULTS: The mean body weight of the invagination group became statistically less than the laparotomy and sham groups at 7 and 21 days. The mean weight of the peritesticular fat pad from the inv group was also significantly less than from the sham group but not different from the lap group. CONCLUSION: Gastric greater curvature invagination significantly decreases weight in rats. PMID- 16469220 TI - A comparison of wedge and needle hepatic biopsy in open bariatric surgery. AB - BACKGROUND: Morbidly obese patients, despite normal laboratory tests and no clinical evidence of liver disease, present a high prevalence of hepatic histological changes. Liver biopsy is able to provide the diagnosis, staging and assessment of follow-up of hepatic disease, thus helping to define clinical management. There is no agreement on which biopsy technique provides better material for analysis. Considering that subcapsular fibrosis is a common finding, sampling from deeper sites is necessary to achieve an adequate histological assessment. METHODS: A study was done in 264 consecutive morbidly obese patients who underwent open Roux-en-Y gastric bypass between July 2001 and Sept 2004, in whom an intraoperative liver biopsy was taken. The first 107 were wedge biopsies, and the last 157 were needle biopsies. The histological degree of steatosis, presence of fibrosis and adequacy of material from the 2 biopsy techniques were compared. RESULTS: Degree of steatosis in both sampling techniques showed no statistical difference (P=0.132). The presence of fibrosis in wedge biopsies (46.1% fibrosis, n 41) was significantly higher than in needle biopsies (13.7% fibrosis, n 20), P<0.001. As expected, sample size of needle biopsies was smaller than that obtained by the wedge technique (P<0.001), but there was no difference in the quality of material obtained (P=0.95). CONCLUSION: Needle biopsies were as effective as wedge biopsies in assessing the degree of steatosis in morbidly obese patients. More important, the presence of subcapsular fibrosis in needle biopsies was less than in wedge biopsies, suggesting an adequate tissue sample by the less invasive technique. PMID- 16469221 TI - Prevalence and predictive factors of non-alcoholic steatohepatitis (NASH) in morbidly obese patients undergoing bariatric surgery. AB - BACKGROUND: In patients with morbid obesity selected for bariatric surgery, previous studies have shown a prevalence of NASH varying from 2.6% to 91%. The prevalence of NASH and extensive fibrosis were studied in a prospective cohort of patients with morbid obesity requiring bariatric surgery, to identify predictive factors of NASH. METHODS: From July 01 to Sept 02, every patient requiring bariatric surgery had a liver biopsy. The diagnosis of NASH was established using Lee's criteria. RESULTS: 92 patients (85 women, age 38 +/- SEM 11 years) were analyzed. Mean BMI was 45.7 +/- 5.1 kg/m2. 35 patients had lobular inflammation. 9 patients had steatosis associated with lobular necrotic and inflammatory foci and ballooning degeneration or pericellular fibrosis. No cirrhosis or extensive fibrosis was evidenced. The prevalence of NASH in this population was 9.8%. Waist/hips ratio and BMI were independent predictors of lobular inflammation, but only BMI was an independent factor of NASH in multivariate analysis. CONCLUSION: In this prospective cohort of patients at bariatric surgery, the prevalence of NASH was 9.8%. BMI was the only predictive factor for NASH. PMID- 16469222 TI - Melanocortin-4 receptor gene and complications after gastric banding. AB - BACKGROUND: Mutations of the melanocortin-4 receptor (MC4R) gene are associated with up to 5.8% of monogenetic causes of obesity. Correlations between defects in MC4R and complications after laparoscopic adjustable gastric banding (LAGB) have recently been reported. The aim of this study was to investigate whether in our patient population band-associated complications can be correlated with MC4R defects, which in turn could be a contraindication for gastric banding. METHODS: Of 370 morbidly obese patients operated between Dec 1996 and May 2004 with LAGB, 37 required re-operation, by re-banding or biliopancreatic diversion/duodenal switch for band-associated complications. Genomic DNA was extracted from leucocytes of these 37 patients using standard methods. The entire MC4R-gene was amplified by PCR and sequenced on an ABI Prism 3100 automated DNA sequencer. Any detected mutation or polymorphism was verified utilizing a high-fidelity proofreading polymerase. RESULTS: No mutation was seen in 35 patients (95%). The polymorphism Ile251Leu (A1144C) which was found in one patient is known not to be associated with obesity. A silent mutation Ile198Ile (C594T) was found in another patient. Based on published data, approximately 12 patients of the 37 would have been expected to carry one of the known obesity-associated MC4R mutations, but none of these was found. CONCLUSION: In our patient material, we could not confirm the observation previously published that MC4R defects are associated with a higher complication rate following LAGB. Thus, we do not recommend routine general screening for MC4R defects before LAGB. PMID- 16469223 TI - The objective of psychological evaluation in the process of qualifying candidates for bariatric surgery. AB - Psychosocial and behavioral variables play an important role in both the development and treatment of obesity. Therefore, in the process of qualifying the patient for bariatric surgery, it is necessary to professionally evaluate his/her psychological state. Such evaluation is very helpful in the identification of factors potentially disturbing the effectiveness of the treatment. Clinical interviews with a group of 80 patients were conducted by a psychologist in the pre- and post-surgical period. The qualitative analysis of the interviews led to the identification of the major elements which should become the object of psychological evaluation in the process of qualifying patients for bariatric surgery. Conducting a clinical interview comprising these elements allows one to evaluate their potential influence on the process of surgical treatment of obesity and to provide optimal psychological support for the patient before and after the surgery. PMID- 16469224 TI - Intracolonic penetration of the laparoscopic adjustable gastric banding tube. AB - The laparoscopic adjustable gastric band (LAGB) has become a common, usually successful treatment for morbid obesity. Complications occur in 10-20%, most commonly band slippage and port system problems, especially infection of the port area. A rare complication is penetration of the band into the stomach. We report the penetration of the tube connecting the port system with the gastric band, into the colon 1 year after treatment of a port infection. PMID- 16469225 TI - Detachment of the connecting tube from the port and migration into jejunal wall. AB - After laparoscopic adjustable gastric banding (LAGB), early recognition and treatment of uncommon complications are important. A 36-year-old man who had undergone LAGB presented at our clinic with weight gain of 14 kg during the prior 6 months. During investigation, detachment of the connecting tube from the port and migration of this tube into the jejunal wall were found. The detached port was replaced with a new port, and laparoscopically-assisted jejunorraphy was performed. PMID- 16469246 TI - [The understanding of western medicine on disease in the 18th century.] AB - The concept of modern disease is formed by organic pathology, which was born in the 18(th) century and opened the studies in the history of taking disease as a core. This article expounds the level of western medicine on disease in the 18(th) century by discussing the formation of organic pathology, classification of disease, and method of diagnosis. PMID- 16469226 TI - Late sigmoid colon internal herniation into the jejuno-jejunostomy mesenteric defect after laparoscopic Roux-en-Y gastric bypass. AB - A 49-year-old female with morbid obesity (BMI 42) underwent laparoscopic Roux-en Y gastric bypass (LRYGBP). 10 months after the operation, she presented to the hospital with intermittent mid-abdominal pain. An internal hernia of the sigmoid colon through a mesenteric defect of the jejuno-jejunostomy was found. Although small bowel internal herniation has been widely documented, the finding of large bowel internal herniation has not been previously reported. Maintaining a high index of suspicion and a low threshold for urgent intervention are required when evaluating patients with vague abdominal complaints after LRYGBP. PMID- 16469247 TI - [Selection of medical talents under the influence of imperial examination system in the Tang Dynasty.] AB - The Tang Dynasty was an important period in which China's ancient imperial examination system was formed and perfected. The medical examination at that time was influenced by imperial examination system, too. Medical talents were chosen according to a clear regulation of exam subjects imitating "Examination based on Imperial College law", "Ming Jing" and "Ming Fa" in the Tang Dynasty. The ways of choosing sheng tu, gong ju, zhi ju, dai zhao were also used for choosing medical talents, and the practical subjects of medical examination and qualified standard were also made, which can be used as a reference for TCM education, examination, selecting and checking of TCM talents today. PMID- 16469248 TI - [Clinical application of several Taoist magical pellets in the Tang Dynasty.] AB - Alchemy is one the arts of Taoist practice. Clinical application of arsenics, mercury, lead, and alloys brought out by alchemy is the primal effect on TCM clinic in the Tang Dynasty. Their chemical compounds such as arsenides, realgar, silver mercuric alloys, mercuric chlorides, all played an important role in TCM clinical practice in the Tang Dynasty, exerting effects on later ages until today. PMID- 16469249 TI - [Investigation on bei die.] AB - Overseas doctors hold that depression has the symptom of "inappropriate guilt" and TCM has no discussions and treatment on it. This article discusses the origin and development, etiology and pathogenesis, therapeutic principles, prescription of Bei Die, revealing that TCM has had quite comprehensive record on the symptoms and treatment of "inappropriate guilt." PMID- 16469250 TI - [Establishment and development of Xuan Fu (sweat pore) Theory.] AB - Xuan Fu (sweat pore) theory was derived from Huangdi Nei Jing (Canon of Internal Medicine), formulated by Liu Wan-su in his Su Wen Xuan Ji Yuan Bing Shi (Pattern of Mysterious Mechanism of Pathogenesis in Plain Questions) in the Jin Dynasty, and perfected by doctors later. The article introduces the establishment and development of Xuan Fu (sweat pore) theory from its concept and function, pathology for its stasis, therapeutical principles and treatment for opening Xuan Fu, its special theory and modern exploration in its essence. PMID- 16469251 TI - [Influence factors of culture for oint selection in ancient time.] AB - Most original acupuncture and moxibustion prescriptions in ancient times were stemmed from clinical experience, seldom influenced by theories. Acupoints selected in ancient prescriptions were influenced by many factors with the development of TCM theory, continuous infiltration of philosophic and religious ideology, and miscopying and misunderstanding of literature. Those are the very reasons that cause misunderstanding of ancient acupuncture and moxibustion prescriptions. PMID- 16469252 TI - [The Whole Story of The Endeavor for Legalization of Chinese Medicine Education in The Republican peitod of China.] AB - During the Republic an period of China, the inhibition and repulsion of Chinese medicine from the government and Westernized factions hadn't been ceasing all the while. In the meantime, the Chinese medicine education suffered unprecedented devastation and loss. In order to strive for the legalization of Chinese medicine education, the circle set up some Chinese medicine schools and brought up many persons with ability in this field. PMID- 16469253 TI - [Evolution of the definition of transient ischemic attack (TIA).] AB - The definition of TIA began with one-hour cutoff in 1950s and established with 24 hours cutoff during 1960s and 1970s. During the period when no imaging techniques could help to differentiate TIA from ischemic stroke, such definition could only contribute to clinical classification, with inevitable shortcomings. With the development of imaging technique and thrombolysis therapy, TIA has become a good opportunity for identifying and managing ischemic cerebrovascular diseases. The New TIA definition employs the one-hour cutoff with emphasis on no brain lesion evidence. The new definition may help to identify and manage ischemic cerebrovascular diseases earlier. Revision of the definition of diseases calls for the combination of the development of medical science and clinical practice. PMID- 16469254 TI - [History of taxonomy of Headache.] AB - It takes a long history for human being to understand headache disorder clearly. Fifteen years after its original edition was published, a revised International Classification of Headache Disorder has been unveiled by the International Headache Society (HIS) in 2003, thus, a clear operationalized standard for the headache taxonomy and diagnosis for headache is available. This will galvanize headache community and stimulate the development of headache research. By reviewing the history of headache taxonomy in China and western countries, historical experiences may contribute to make further advances not only in the headache research field, but also in the whole area of medical science. PMID- 16469255 TI - [The origin of Red Cross and its history in China.] AB - This article discusses the origin of Red Cross and its history and general condition in China, and reviewes and analyzes that Red Cross is a lofty project for human being. On October 26(th) in 1863, Henry Dunant, the founder of Red Cross, set up the rescue organization which undertook the load of rescuing and protected people's lives and healthy. Red Cross actively performs the responsibility of rescue, efficiently develops series of movement, and makes great contribution to our country's reform and development, our nation's stability and world's peace. PMID- 16469257 TI - [In Process Citation] PMID- 16469256 TI - [The factors influencing the spread of Chinese herbal medicine as reflected in the application of Sanchi (Radix Notoginseng).] AB - The curative effect of medicine is the principal one among so many factors that influence its spread. As an effective blood-stanching herb, Sanchi (Radix Notoginseng) is approved in nearly 20 years and is cultivated widely with its sources ensured. PMID- 16469258 TI - [A brief discussion on western public health history.] AB - The study of public health history, bound to follow the development of public health facilities, is an important branch among all the studies of western social history of medicine. The development of public health history has experienced a course from optimism to objectivity as public health facilities. The study of public health history embraces three major aspects, hygiene, social welfare and philanthropy, those all have a distinct sociality and reality. PMID- 16469260 TI - [In Process Citation] PMID- 16469261 TI - [In Process Citation] PMID- 16469259 TI - [In Process Citation] PMID- 16469262 TI - [In Process Citation] PMID- 16469263 TI - [In Process Citation] PMID- 16469264 TI - Diabetic peripheral neuropathy: an update on pathogenesis and management. PMID- 16469265 TI - Imaging of the brain and cerebral vasculature in patients with suspected stroke: advantages and disadvantages of CT and MRI. AB - Although neuroimaging remains the foundation for the diagnosis of cerebrovascular disease, ongoing technologic advances have now opened up new frontiers for stroke evaluation and treatment. Neuroimaging studies can provide crucial information regarding tissue injury (size, location, and degree of reversibility of ischemic injury as well as presence of hemorrhage), vessel status (site and severity of stenoses and occlusions), and cerebral perfusion (size, location, and severity of hypoperfusion). This information can be combined to identify patients with salvageable penumbral tissue who may benefit most from acute therapies. The multimodal combinations of advanced imaging techniques, particularly in the realm of CT and MRI, have emerged as the most promising noninvasive approaches to acute stroke evaluation. PMID- 16469266 TI - Latest advances in intracerebral hemorrhage. AB - Despite the fact that intracerebral hemorrhage (ICH) is the deadliest and least treatable of all stroke subtypes, historically researchers have directed most of their efforts toward ischemic strokes. However in the past few years this tendency has been changing, and several studies are showing very interesting results that allow us to believe that in the following years ICH management will change dramatically, paralleling the recent revolution that ischemic stroke treatment experienced in the past decade. Studies offering a better understanding of risk factors, pathophysiology, and treatment will help in primary and secondary prevention and also in developing therapeutic strategies to reduce brain damage. This review comments on some of the most relevant publications during the past year in any field related to ICH. PMID- 16469268 TI - Advances in treatment of acute ischemic stroke. AB - Stroke carries a severe toll in terms of loss of life and disability for patients and their families. Until 10 years ago, physicians, and in particular neurologists, had a conservative, non-aggressive approach to this devastating disease. The advent of thrombolytic therapy not only proved that acute ischemic stroke is treatable, but also that early reperfusion can dramatically change the outcome of acute stroke patients. As a result of these trials, intravenous (IV) tissue plasminogen activator (t-PA) has been approved for treatment of acute ischemic stroke within 3 hours after symptom onset in the United States, Canada, Australia, and the European Union. The near future is extremely promising. Imaging modalities, such as diffusion- and perfusion-weighted images, as well as CT perfusion and CT angiography, to better select patients for treatment are now routinely performed in most academic medical centers. Novel IV and intra-arterial (IA) agents have been developed and tested. Emerging therapies will soon be available to increase the therapeutic windows for thrombolysis both by better screening patients using MRI or CT and by new IV and IA treatments. Several multicenter controlled trials in both imaging-guided decisions and therapeutic agents are either completed or being performed. We review data on advancement in imaging and treatment of acute ischemic stroke, in particular focusing on pharmacologic and mechanical IA thrombolysis. PMID- 16469267 TI - Heart diseases and stroke. AB - A heterogeneous group of heart diseases, varying by cardiac anatomy and physiology, are responsible for a variety of embolic materials that result in cerebral and systemic embolism. Atrial fibrillation accounts for nearly half of all cardioembolic stroke, particularly in the elderly, but in the young congenital septal abnormalities such as patent foramen ovale play a more important role. Therapeutic strategies include stabilization or treatment of the underlying heart disease and prevention of embolism. Oral anticoagulant therapy is indicated for many but decision making on balancing the benefits versus risks can be difficult. New endovascular therapies and devices have become available and await clinical trials that allow them to be compared with existing medical therapies. PMID- 16469269 TI - Does ACE inhibitor therapy delay onset and progression of cardiac dysfunction in duchenne muscular dystrophy? PMID- 16469270 TI - Amyotrophic lateral sclerosis associated with mutations in the CuZn superoxide dismutase gene. AB - This review highlights recent epidemiologic, clinical-genetic, and neurochemical advances in our understanding of sporadic amyotrophic lateral sclerosis (ALS) and their relationships to familial ALS caused by superoxide dismutase (SOD1) gene mutations. It is of fundamental importance to recognize that ALS is a biologically heterogeneous syndrome in which genetics, environment, and aging are inter-related. The discovery of mutations in the SOD1 gene is the greatest breakthrough in ALS research since Charcot's description of the disorder, but the putative toxic gain of function of mutant SOD1 remains elusive despite intense research. Currently, two dominant theories for the pathogenesis of SOD1 mutations exist: specific protein cytotoxicity and protein aggregation. Mutant SOD1 interacts specifically with neurofilament-light chain mRNA and the dynein/dynactin complex, suggesting that cytoskeletal defects and axonal transport are key players. In addition, mutant SOD1 protein has increased propensity to form aggregate-prone monomers, and the degree of instability correlates inversely with length of survival; therefore, increased propensity to aggregate may be the unifying common denominator for the 119 diverse SOD1 mutations. PMID- 16469271 TI - Challenges for gene therapy for muscular dystrophy. AB - Gene therapy for muscular dystrophy represents a promising avenue of pursuit for a disease with a limited repertoire of treatment. Recent successes in the research arena using adeno-associated viral vectors should accelerate the movement of gene-based therapeutics for muscle disorders into the clinic. Nevertheless, significant challenges remain before gene therapy can deliver on the promises avowed by early pioneers of the field. This review examines recent progress and the hurdles remaining to achieve gene-based treatment therapies for muscular dystrophy. PMID- 16469272 TI - Immune-mediated autonomic neuropathies. AB - Improved recognition and availability of noninvasive testing of autonomic disorders has prompted a better understanding of disease mechanisms of some disease forms, especially potentially treatable immune-mediated autonomic neuropathies. Development is acute, subacute, or less commonly chronic. Autonomic involvement is common and an important cause of morbidity and mortality in Guillain-Barre syndrome. Acute autonomic neuropathy can affect parasympathetic, sympathetic, and enteric nerves or neurons and is associated with antibodies to ganglionic nicotinic acetylcholine receptors. These antibodies appear to be causative based on a rabbit immunization model and serum transfer studies from patients and animals. Other important immune autonomic disorders discussed include Lambert-Eaton myasthenic syndrome, some forms of orthostatic intolerance, chronic autonomic neuropathy, and Sjogren syndrome. Paraneoplastic autonomic disorders are clinically indistinguishable and associated with various overlapping antibody associations, including anti-Hu (ANNA-1), ganglionic acetylcholine receptors, CRMP-5, and PCA-2. PMID- 16469273 TI - Hereditary spastic paraplegia. AB - The hereditary spastic paraplegias (HSPs) comprise a large group of inherited neurologic disorders. HSP is classified according to the mode of inheritance, the HSP locus when known, and whether the spastic paraplegia syndrome occurs alone or is accompanied by additional neurologic or systemic abnormalities. Analysis of 11 recently discovered HSP genes provides insight into HSP pathogenesis. Hereditary spastic paraplegia is a clinical diagnosis for which laboratory confirmation is sometimes possible, and careful exclusion of alternate and co-existing disorders is an important element in HSP diagnosis. Treatment for HSP is presently limited to symptomatic reduction of muscle spasticity, reduction in urinary urgency, and strength and gait improvement through physical therapy. Prenatal genetic testing in HSP is possible for some individuals with the increasing availability of HSP gene analysis. PMID- 16469274 TI - Current therapies for neuromuscular manifestations of paraneoplastic syndromes. AB - The neuromuscular manifestations of paraneoplastic disorders result in diverse syndromes that may also occur in patients without cancer. In general, treatment of these disorders is the same whether or not there is an underlying malignancy. However, when the disorder is believed to be paraneoplastic, the main concern should be prompt detection and treatment of the tumor, as this has been shown to offer the best chance for neurologic stabilization or improvement. The paraneoplastic neuromuscular disorders can be divided into two main categories: those that are directly mediated by antibodies and those that are believed to result from other immune-mediated mechanisms, including cytotoxic T-cell responses with or without association with specific antibodies. For disorders in which the antibodies are pathogenic, therapy is aimed at removing the antibodies. For the other disorders, adjuvant therapies are for the most part empiric and include a variety of immunosuppressant and immunomodulatory agents. PMID- 16469275 TI - [Electrolyte disorders following colonic cleansing for imaging studies]. AB - BACKGROUND AND OBJECTIVE: An adequate bowel cleansing is needed prior to radiologic and endoscopic procedures. However, it may have a number of adverse effects, including abnormalities of calcium-phosphorus homeostasis. PATIENTS AND METHOD: This was an observational prospective study in a hospital practice setting. We included consecutive inpatients (n = 47) subjected to a barium enema or colon endoscopy. Prior cleansing was done as indicated by the attending physician by using a low-salt oral poliethylenglicol (PEG) solution, oral sodium phosphate or a phosphate-containing enema. RESULTS: PEG solution frequently caused mild increases in serum sodium, and decreases in serum potassium. Oral phosphate caused a significant increase in serum phosphorus and parathormone concentrations, whereas it decreased serum calcium. Mild hyperphosphatemia was found in 57% of cases, and hypocalcemia in 36%. Phosphate enema also increased serum phosphate, causing mild hyperphosphatemia (33% cases). Although in the whole subgroup of enema-treated patients there were no significant changes in serum calcium, mild hypocalcemia was found in 27% cases. CONCLUSIONS: Bowel cleansing procedures, particularly those using oral phosphate salts, frequently induce hyperphosphatemia and other abnormalities in serum electrolytes. Although usually transitory and without overt clinical consequences, clinicians should be aware of this potential risk, especially in elderly patients and those with impaired renal function. PMID- 16469276 TI - [Quality of care indicators related to premature readmission and death after hospitalization for heart failure]. AB - BACKGROUND AND OBJECTIVE: To determine wether the quality of care criteria applied and the treatment provided to patients hospitalized after congestive heart failure were appropriate to reduce rates of premature readmission and death. PATIENTS AND METHOD: We analyzed the epidemiologic, clinical and quality of care data proposed by 3 international organizations: JCAHO, AHA/ACC and ACOVE Project. The dependent variable was defined as readmission or death during the 30 days after discharge. A multivariate analysis was made using multiple binary logistic regression of the parameters of quality of care and treatment appropriateness. RESULTS: 225 hospital discharge records were analyzed. There were 21 readmissions and 3 deaths (i.e., 24 cases [10.7%] with a positive dependent variable). 162 records (72%) corresponded to patients aged 65 years and over, who presented a total of 18 (8%) readmissions or premature deaths. A positive association between readmission or premature death was found with regard to 2 variables: appropriate treatment with beta-blockers (odds ratio [OR] = 0.34) and the Charlson index (OR = 3,79 for score of 3 or more vs. score of 2 or less). In the case of patients aged 65 years and over the same 2 variables were positively associated, with OR similar to those cited (OR = 0.31 and 3.21, respectively). No association was found between premature readmission or death and the overall evaluation of the criteria referred to by AHA/ACC, JCAHO or the ACOVE Project. CONCLUSIONS: Premature readmission or death of patients with heart failure is more determined by the characteristics of the clinical state of patients (the Charlson comorbidity index) and by the appropriateness of the treatment applied (treatment with beta-blockers) than by the accomplishment of quality of care criteria as proposed by the cited scientific organizations. PMID- 16469277 TI - [Cardiac surgery in octogenarians: current situation and future perspectives]. AB - BACKGROUND AND OBJECTIVE: Increasing life expectancy makes cardiac surgery in octogenarians not very uncommon. We evaluated the characteristics and outcomes of patients aged over 80 undergoing cardiac surgery in our centre. PATIENTS AND METHOD: We retrospectively analyzed preoperative risk factors, in-hospital morbimortality, and long term survival in 120 patients aged 80 years--mean age (standard deviation): 81.53 (1.83)--who underwent cardiac surgery between 2000 and 2005, in comparison with 2425 younger patients referred to our center during the same period. RESULTS: Preoperatively, octogenarians presented arterial hypertension, renal failure, unstable angina, left main coronary stenosis and New York Heart Association (NYHA) functional class III or IV more frequently than younger patients. Combined cardiac surgery procedures (valvular plus coronary) were also significatively higher (20% vs. 9.85%). Hospital mortality (15.8%) was twice as higher as in the non octogenarian population and postoperative intensive care units and hospital stay was longer. 45% suffered some postoperative complication. In the multivariate analysis, urgent procedure, mitral valve surgery and presentation of any postoperative complication were identified as predictors of mortality, and female sex was identified as a risk factor for postoperative morbidity. Survival at one, 3 and 5 years was 91.20%, 80.73% and 69.20%, respectively. 98.7% of the patients were in I-II NYHA functional class and 92.5% satisfied with overall surgery outcome when they were interviewed. CONCLUSIONS: Despite more preoperative co-morbidity and worse functional class, cardiac surgery in octogenarians can be performed with an acceptable mortality. Our patients have a satisfactory quality of life and would undergo a cardiac surgery procedure again. Specific measurements in preoperative and immediate postoperative periods, in order to reduce higher morbidity rates, are advisable. PMID- 16469278 TI - [Purgative with high sodium phosphate contents: efficacious but not so safe]. PMID- 16469279 TI - [Legionnaires disease. From the first case to the outbreak]. PMID- 16469280 TI - [The threat of avian influenza human pandemic]. PMID- 16469281 TI - [Prevalence of Legionella in cooling towers in the Community of Madrid]. PMID- 16469282 TI - [Ignoring familiar decision for increasing organ donations?]. PMID- 16469284 TI - [Simple closure and Helicobacter pylori eradication in perforated ulcer]. PMID- 16469286 TI - [Too many theories of health promotion?]. PMID- 16469285 TI - [Atypical presentation of MEN 1 syndrome]. PMID- 16469287 TI - [Fournier's gangrene with fatal progress by a fish bone]. PMID- 16469288 TI - Genetic variability of Brazilian populations of Lymnaea columella (Gastropoda: Lymnaeidae), an intermediate host of Fasciola hepatica (Trematoda: Digenea). AB - In Brazil, Lymnaea columella is the most important intermediate host of Fasciola hepatica, the etiological agent of fasciolosis, which is a parasitic disease of veterinarian and human importance. Random amplified polymorphic DNA (RAPD) was used to investigate the genetic variability within and among nine Brazilian populations of L. columella comprising 205 individuals. A number of four primers were used for analysis of molecular variance (AMOVA). Out of 83 RAPD markers, 63 (76%) were polymorphic and revealed 119 unique RAPD profiles. The levels of genetic variability found in the populations were low and most of the genetic variation was interpopulational (81.6%) when compared to intrapopulational variability (18.4%). These results are in accordance with the dynamics and distribution of the populations analyzed. PMID- 16469289 TI - [Acute abdomens of parasitic origin: retrospective analysis of 135 cases]. AB - OBJECTIVE: To assess acute abdomens incidence of parasitic origin at Yaounde (Cameroon) and evaluate their different management modalities. MATERIALS AND METHODS: Retrospective study from January 1973 to December 2002 of patients managed at Department of Surgery, Central Hospital, Yaounde (Cameroon). RESULTS: Among 3464 acute abdomens managed by laparotomy during this period, 135 patients (3.9%) had a parasitic origin. Ninety-seven patients (79%) were operated on before 1990. Among these 135 patients, 63 (47%) had peritonitis secondary to liver amoebic abscess intraperitoneal rupture, 24 (18%) had acute intestinal obstruction due to an ascaridioma, 21 (15%) had appendicitis of parasitic origin, 15 (11%) had amoebic typhlitis, 4 (3%) had pancreatitis and/or angiocholitis caused by the obstruction of ampulla of Vater by an adult ascaris worm, 4 (3%) had intestinal perforation by ascaris, and 4 (3%) had intussusception (tricocephalus). CONCLUSIONS: Incidence of abdominal emergencies of parasitic origin is unfrequent but not rare at Central Hospital, Yaounde, Cameroon. This incidence is decreasing over time. However, liver amoebic abscess intraperitoneal rupture is the most common cause of peritonitis and is still associated with postoperative high mortality rate. Preoperative diagnosis could lead to non operative management that may be associated with better prognosis. PMID- 16469290 TI - Expression of a novel P275L variant of NADH:cytochrome b5 reductase gives functional insight into the conserved motif important for pyridine nucleotide binding. AB - The clinical disorder of recessive congenital methemoglobinemia (RCM, OMIN 250800) is associated with mutations in NADH:cytochrome b5 reductase (cb5r) and manifests as cyanosis from birth. Screening a cyanotic infant indicated elevated methemoglobin levels and decreased cb5r activity suggesting RCM. Sequencing the DIA1 gene encoding cb5r revealed a novel mutation, C27161T (NCBI accession number: NT_011520), resulting in replacement of proline at amino acid 275 with leucine (P275L). To understand how this mutation would affect cb5r's function, the P275L variant was expressed in a heterologous expression system and spectroscopic, thermodynamic, and thermostability studies were performed. The leucine substitution at residue 275 was found to significantly decrease the affinity towards the physiological reducing substrate, NADH, without affecting the activity of the P275L variant. From the rat model, residue 275 is predicted to be part of a conserved "CGPPPM" motif important for the binding and correct positioning of the NADH reducing substrate. Thus P275 influences the interaction with NADH which was confirmed by the change in affinity towards the physiological reducing substrate. PMID- 16469291 TI - The LTR promoter of the rat oncomodulin gene is regulated by cell-line specific accessibility in the LTR U3 region. AB - By germline insertion, a long terminal repeat (LTR) of an intracisternal A particle type IAP retrovirus has overtaken the transcriptional control of the rat oncomodulin (OM) gene, which codes for a high affinity Ca2+-binding protein with modulatory capacity. In order to get insights into regulatory mechanisms of LTR directed OM gene expression we tested promoter activity of this LTR by transient transfection of transformed rat fibroblasts with this sequence placed 5' of the human growth hormone hGH reporter gene. The OM LTR is a strong promoter but does not follow an expression pattern similar to the one of the OM gene. Genomic sequencing showed a good correlation between CpG hypomethylation in the OM LTR and OM transcription among various cell lines and tissues. DNase I mapping of a 18 kb fragment containing the OM gene and 5' flanking sequences revealed cell line specific hypersensitivity sites located within the U3 region of the LTR element. Several cis-elements in the OM LTR promoter exhibiting cell-line specific occupancy were identified by in vivo DMS-footprinting. Detailed analysis of protein interactions with two such sequence elements in vitro revealed binding of ubiquitously expressed nuclear factors within an AP-1 (activator protein 1) and a intracisternal A-particle upstream enhancer recognition sequence. Protein occupancy to the latter sequence is significantly reduced by CpG methylation. These results indicate that cell-line specificity of OM expression is dictated by factor accessibility to the LTR promoter. PMID- 16469292 TI - Wnt-10b promotes differentiation of skin epithelial cells in vitro. AB - To evaluate the role of Wnt-10b in epithelial differentiation, we investigated the effects of Wnt-10b on adult mouse-derived primary skin epithelial cells (MPSEC). Recombinant Wnt-10b protein (rWnt-10b) was prepared using a gene engineering technique and MPSEC were cultured in its presence, which resulted in morphological changes from cuboidal to spindle-shaped and inhibited their proliferation. Further, involvement of the canonical Wnt signal pathway was also observed. MPSEC treated with rWnt-10b showed characteristics of the hair shaft and inner root sheath of the hair follicle, in results of Ayoub Shklar staining and immunocytochemistry. Further, the cells expressed mRNA for differentiated epithelial cells, including keratin 1, keratin 2, loricrin, mHa5, and mHb5, in association with a decreased expression of the basal cell marker keratin 5. These results suggest that Wnt-10b promotes the differentiation of MPSEC. PMID- 16469293 TI - Effect of Nd3+ ion on carboxylation activity of ribulose-1,5-bisphosphate carboxylase/oxygenase of spinach. AB - Neodymium (Nd), as a member of rare earth elements, proved to enhance the photosynthesis rate and organic substance accumulation of spinach through the increase in carboxylation activity of Rubisco. Although the oxygenase activity of spinach Rubisco was slightly changed with the Nd(3+) treatment, the specific factor of Rubisco was greatly increased. It was partially due to the promotion of Rubisco activase (R-A) activity but mainly to the formation of Rubisco-Rubisco activase super-complex, a heavier molecular mass protein (about 1200kD) comprising both Rubisco and Rubisco activase. This super-complex was found during the extraction procedure of Rubisco by the gel electrophoresis and Western-blot studies. The formation of Rubisco-R-A super-complex suggested that the secondary structure of the protein purified from the Nd(3+)-treated spinach was different from that of the control. Extended X-ray absorption fine structure study of the 'Rubisco' purified from the Nd(3+)-treated spinach revealed that Nd was bound with four oxygen atoms and two sulfur atoms of amino acid residues at the Nd-O and Nd-S bond lengths of 2.46 and 2.89A, respectively. PMID- 16469294 TI - Sequestering ErbB2 in endoplasmic reticulum by its autoinhibitor from translocation to cell surface: an autoinhibition mechanism of ErbB2 expression. AB - ErbB2 is differentially overexpressed in tumor versus host tissues, suggesting that an autoregulation mechanism may modulate the expression of ErbB2 and control cell growth. A truncated ErbB2 extracellular domain, herstatin has been shown to bind to ErbB2 and inhibit the growth of tumor cells expressing ErbB2. In the present study, the interaction of herstatin and ErbB2 in vivo was observed by confocal microscopy. The aggregation of ErbB2 and herstatin was found in endoplasmic reticulum (ER). The decrease of ErbB2 on the cell surface was accompanied with the increased colocalization of ErbB2 and herstatin in the cytoplasm, suggesting that the formation of ErbB2/herstatin complex may prevent transit from ER to cell surface of ErbB2. The formation of ErbB2 and herstatin complex was further confirmed by immunoprecipitation. The results demonstrate that sequestering ErbB2 molecules intracellularly by herstatin may be a possible mechanism of the cell growth inhibition. PMID- 16469295 TI - The effects of blood feeding and exogenous supply of tryptophan on the quantities of xanthurenic acid in the salivary glands of Anopheles stephensi (Diptera: Culicidae). AB - Xanthurenic acid (XA), produced as a byproduct during the biosynthesis of insect eye pigment (ommochromes), is a strong inducer of Plasmodium gametogenesis at very low concentrations. In previous studies, it was shown that XA is present in Anopheles stephensi (Diptera: Culicidae) mosquito salivary glands and that during blood feeding the mosquitoes ingested their own saliva into the midgut. Considering these two facts together, it is therefore likely that XA is discharged with saliva during blood feeding and is swallowed into the midgut where it exerts its effect on Plasmodium gametocytes. However, the quantities of XA in the salivary glands and midgut are unknown. In this study, we used high performance liquid chromatography with electrochemical detection to detect and quantify XA in the salivary glands and midgut. Based on the results of this study, we found 0.28+/-0.05 ng of XA in the salivary glands of the mosquitoes, accounting for 10% of the total XA content in the mosquito whole body. The amounts of XA in the salivary glands reduced to 0.13+/-0.06 ng after mosquitoes ingested a blood meal. Approximately 0.05+/-0.01 ng of XA was detected in the midgut of nonblood fed An. stephensi mosquitoes. By adding synthetic tryptophan as a source of XA into larval rearing water (2 mM) or in sugar meals (10 mM), we evaluated whether XA levels in the mosquito (salivary glands, midgut, and whole body) were boosted and the subsequent effect on infectivity of Plasmodium berghei in the treated mosquito groups. A female specific increase in XA content was observed in the whole body and in the midgut of mosquito groups where tryptophan was added either in the larval water or sugar meals. However, XA in the salivary glands was not affected by tryptophan addition to larval water, and surprisingly it reduced when tryptophan was added to sugar meals. The P. berghei oocyst loads in the mosquito midguts were lower in mosquitoes fed tryptophan treated sugar meals than in mosquitoes reared on tryptophan treated larval water. Our results suggest that mosquito nutrition may have a significant impact on whole body and midgut XA levels in mosquitoes. We discuss the observed parasite infectivity results in relation to XA's relationship with malaria parasite development in mosquitoes. PMID- 16469296 TI - Nuclear translocation and overexpression of GAPDH by the hyper-pressure in retinal ganglion cell. AB - To investigate the effect of hyper-pressure on retinal ganglion cells (RGC-5), RGC-5 cells were exposed to an ambient hydrostatic pressure of 100 mmHg. Upon treatment, the proliferation of RGC-5 cells was inhibited and neuronal apoptosis was detected by specific apoptosis marker TUNEL (terminal deoxynucleotidyl transferase dUTP nick-end labeling). To probe into the mechanism mediating the apoptosis of RGC-5 cells in 100 mmHg, protein profile alterations following hyper pressure treatment were examined using two-dimensional gel electrophoresis (2-DE) followed by MALDI-TOF. Out of the 400 protein spots of RGC-5 cells detected on 2 DE gels, 37 differentially expressed protein spots were further identified using in gel tryptic digestion and mass spectrometry. Among these proteins, glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) was significantly expressed 10 times more in 100 mmHg than in normal pressure. The accumulation of GAPDH in the nucleus and its translocation from the cytosol to the nucleus in 100 mmHg were observed using a microscope. These results suggest that the hyper-pressure induced apoptosis in RGC-5 cells may be involved with not only the increase of GAPDH expression, but also the accumulation and the translocalization of GAPDH to the nucleus. PMID- 16469297 TI - Microdosimetry for conventional and supra-electroporation in cells with organelles. AB - Conventional electroporation (EP) by 0.1 to 1 kV/cm pulses longer than 100 micros, and supra-electroporation by 10 to 300 kV/cm pulses shorter than 1 micros cause different cellular effects. Conventional EP delivers DNA, proteins, small drugs, and fluorescent indicators across the plasma membrane (PM) and causes moderate levels of phosphatidylserine (PS) translocation at the PM. We hypothesize that supra-EP is central to intracellular effects such as apoptosis induction and higher levels of PS translocation. Our cell system model has 20,000 interconnected local models for small areas of the PM and organelle membranes, small regions of aqueous media, appropriate resting potentials, and the asymptotic EP model. Conventional EP primarily affects the PM, but with a hint of endoplasmic reticulum involvement. Supra-EP can involve all of a cell's membrane at the largest fields. Conventional EP fields tend to go around cells, but supra EP fields go through cells, extensively penetrating organelles. PMID- 16469298 TI - Bridging the gaps in 3D structure of the inositol 1,4,5-trisphosphate receptor binding core. AB - Calcium concentration is strictly regulated in all cells. The inositol 1,4,5 trisphosphate receptor (IP(3)R), which forms a homotetrameric Ca2+ release channel in the endoplasmic reticulum, is one of the key molecules responsible for this regulation. The opening of this channel requires binding of two intracellular messengers, which are inositol 1,4,5-trisphosphate (IP(3)) and Ca2+. To promote the Ca2+-channel gating and release from the endoplasmic reticulum, IP(3) binds to the amino-terminal region of IP(3)R. Recently, the crystal structure of IP(3)R-binding core in complex with its ligand was presented [I. Bosanac, J.R. Alattia, T.K. Mai, J. Chan, S. Talarico, F.K. Tong, K.I. Tong, F. Yoshikawa, T. Furuichi, M. Iwai, T. Michikawa, K. Mikoshiba, M. Ikura, Structure of the inositol 1,4,5-trisphosphate receptor binding core in complex with its ligand, Nature 420 (2002) 696-700; I. Bosanac, H. Yamazaki, T. Matsu ura, T. Michikawa, K. Mikoshiba, M. Ikura, Crystal structure of the ligand binding suppressor domain of type 1 inositol 1,4,5-trisphosphate receptor, Mol. Cell 17 (2005) 193-203]. The space positions of residues 289-301 (segment A), 320 350 (segment B), 373-386 (segment C), and 529-545 (segment D) were not determined by the X-ray crystallography. To bridge these gaps, the computer modeling of physiologically meaningful low-energy 3D structures of the segments A-D of the inositol 1,4,5-trisphosphate receptor has been carried out by using a hierarchical conformational search algorithm combining two approaches: knowledge based homology modeling and ab initio conformational search strategy. The structure analysis suggests a Ca2+-binding site of high affinity formed by residues 296-335, several low-energy regular secondary structure units within the segment B, and a number of hinge regions within the segments A-D, important for the receptor functioning. PMID- 16469299 TI - Marked sexual dimorphism of lacrimal gland peroxidase in hamster: repression by androgens and estrogens. AB - Peroxidase secreted in tears by the lacrimal glands is a marker of secretory activity of these glands and is believed to have an antimicrobial function. We report for the first time a marked sex difference in lacrimal gland (LG) peroxidase in hamsters ( approximately 3.4-fold higher activity in females), which is due to an unusual repression by physiological levels of androgens in males. LG peroxidase activity was markedly induced in a time-dependent manner after gonadectomy in males and also females ( approximately 8- and 2-fold, respectively) and was strongly repressed by androgen treatment in a dose- and time-dependent manner. Estrogen treatment of gonadectomized hamsters could also repress LG peroxidase but not below female levels. These repressions by androgens and estrogens were significantly prevented upon co-treatment with their respective receptor antagonists. Western blotting showed that differences in LG peroxidase specific activity, in different sex hormonal states and treatments were due to changes in the levels of peroxidase protein in LG. A tear peroxidase with a clear sex difference suggests that it might also have other novel function(s) in hamster tears. PMID- 16469300 TI - Antibody-bound amyloid precursor protein upregulates ornithine decarboxylase expression. AB - Alzheimer's disease is a neurodegenerative disorder characterised by extracellular accumulation of the Abeta peptide, derived from the amyloid precursor protein (APP). The function of APP as a cell surface receptor was examined by ligand-mimicking using an antibody against the APP extracellular domain. Alterations in gene expression evoked by antibody-bound APP were analysed using human pathway-finder gene arrays and the largest change in expression levels was found for ornithine decarboxylase (ODC). These results were confirmed by Western blotting which showed even higher upregulation on the protein level. APP knockdown by RNAi verified that upregulation of ODC was APP-mediated. This APP signalling event did not require gamma-secretase cleavage, as it was independent of the presence of presenilin-1 or -2. The induced ODC expression was rapid and biphasic, resembling growth-factor stimulated signalling events. This study shows that antibody-bound APP leads to altered gene expression that may be relevant to AD. PMID- 16469301 TI - Target-mediated drug disposition and dynamics. AB - Nonlinear pharmacokinetics and pharmacodynamics may result from several capacity limited processes and often represent complicating factors in characterizing the pharmacological properties of drugs. Target-mediated drug disposition (TMDD) corresponds to a special case wherein a significant proportion of a drug (relative to dose) is bound with high affinity to a pharmacological target, such that this interaction is reflected in the pharmacokinetic properties of the drug. Dose-dependent effects on apparent pharmacokinetic parameters may manifest, including the steady-state volume of distribution and total systemic clearance. Although a few small molecular weight compounds have been identified to exhibit TMDD, the incidence of TMDD is likely to increase particularly among emerging biotechnology pharmaceuticals. The goal of this commentary is to describe the basic tenets of TMDD and discuss several mathematical modeling approaches for characterizing this phenomenon. Whereas traditional pharmacokinetic/pharmacodynamic models assume that the amount of the drug-target complex is negligible relative to the total amount of drug in the body, integrated mechanism-based models of TMDD incorporate the binding and stoichiometry of drug-target binding. These models may be utilized to infer the time-course of inaccessible system variables, such as the in vivo density of the drug-target complex, and provide a suitable platform for ascertaining the apparent pharmacodynamic implications of TMDD. PMID- 16469302 TI - Reduced pain sensitivity in mice lacking latexin, an inhibitor of metallocarboxypeptidases. AB - Latexin, the endogenous protein inhibitor of the A/B subfamily of metallocarboxypeptidases, is expressed in small nociceptive neurons in sensory ganglia and in a subset of neurons in the telencephalon. In this study, we generated latexin-deficient mice that exhibited increased tail-flick latency compared to wild-type animals upon noxious heat stimulation. The reduced pain sensitivity in the mutants was rescued by the systemic administration of a plant carboxypeptidase inhibitor that inhibits the A/B subfamily of metallocarboxypeptidases. These findings suggest that latexin is involved in the transmission of pain. PMID- 16469303 TI - Gene expression analysis of the development of congenital hydrocephalus in the H Tx rat. AB - To discover candidate genes in the pathogenesis of congenital hydrocephalus, gene arrays were utilized to analyze transcripts from the midbrain region of 5-day-old H-Tx rats; these animals develop hydrocephalus due to closure of their cerebral aqueduct between embryonic day 18 and post-natal day 5. Of the 15,924 transcripts assayed, we detected 47 differentially expressed transcripts representing 23 genes and 24 expressed sequence tags (ESTs); 17 transcripts (7 genes and 10 ESTs) were upregulated and 30 (16 genes and 14 ESTs) were downregulated in the hydrocephalic animals relative to control non-hydrocephalic animals. Seven of these genes, Cck, Nfix, Lgals3, Gsta1, Xdh, Tnf, and Tfpi-2, can be linked to hydrocephalus. In addition, 17 genes that displayed altered expression in our study are not currently known to be associated with the presence or development of hydrocephalus. These results indicate that a relatively few number of transcripts were found to be altered in the development of hydrocephalus in this model. This is the first experiment of its kind to identify changes in gene expression in a congenital model of rodent hydrocephalus that are occurring locally in the area surrounding the cerebral aqueduct. Studies are now needed to examine these candidate genes and their cognate proteins to delineate their role in hydrocephalus. PMID- 16469304 TI - Serum markers of vascular inflammation in dyslipemia. AB - Atherosclerosis is widely recognized as an inflammatory disease because systemic and local inflammatory events mediate all phases of plaque development and progression. Basic and clinical studies have focused on identifying potentially useful markers of inflammation. In this article, we review the inflammatory pathogenesis of atherosclerosis, and highlight recent results of several of the more promising markers of inflammation for cardiovascular risk assessment. Of these markers, the most reliable and accessible for clinical use is currently high-sensitivity C-reactive protein (CRP). At present, most clinical guidelines do not recommend routine measurement of these inflammatory markers. However, these serum markers of vascular inflammation may be useful as an adjunct to lipid screening, especially for patients whose lipid values may not be severely elevated, but who are at intermediate risk according to scoring systems that take into account multiple established risk factors. In addition, since the pleiotropic effects of statins include the inhibition of inflammatory response, serum inflammatory markers could also be useful for monitoring this action. Nevertheless, several issues have to be evaluated before the measurements of inflammatory markers can be used for cardiovascular risk prediction in either clinical practice or in clinical trials evaluating anti-atherosclerotic drugs. PMID- 16469305 TI - Developmental timing in Dictyostelium is regulated by the Set1 histone methyltransferase. AB - Histone-modifying enzymes have enormous potential as regulators of the large scale changes in gene expression occurring during differentiation. It is unclear how different combinations of histone modification coordinate regimes of transcription during development. We show that different methylation states of lysine 4 of histone H3 (H3K4) mark distinct developmental phases of the simple eukaryote, Dictyostelium. We demonstrate that the enzyme responsible for all mono, di and tri-methylation of H3K4 is the Dictyostelium homolog of the Set1 histone methyltransferase. In the absence of Set1, cells display unusually rapid development, characterized by precocious aggregation of amoebae into multicellular aggregates. Early differentiation markers are abundantly expressed in growing set1 cells, indicating the differentiation program is ectopically activated during growth. This phenotype is caused specifically by the loss of Set1 catalytic activity. Set1 mutants induce premature differentiation in wild type cells, indicating Set1 regulates production of an extra-cellular factor required for the correct perception of growth conditions. Microarray analysis of the set1 mutants reveals genomic clustering of mis-expressed genes, suggesting a requirement for Set1 in the regulation of chromatin-mediated events at gene clusters. PMID- 16469306 TI - Evidence for motoneuron lineage-specific regulation of Olig2 in the vertebrate neural tube. AB - Within the motoneuron precursor (pMN) domain of the developing spinal cord, the bHLH transcription factor, Olig2, plays critical roles in pattern formation and the generation of motor neuron and oligodendrocyte precursors. How are the multiple functions of Olig2 regulated? We have isolated a large BAC clone encompassing the human OLIG2 locus that rescues motor neuron and oligodendrocyte development but not normal pattern formation in Olig2(-/-) embryos. Within the BAC clone, we identified a conserved 3.6 kb enhancer sub-region that directs reporter expression specifically in the motor neuron lineage but not oligodendrocyte lineage in vivo. Our findings indicate complex regulation of Olig2 by stage- and lineage-specific regulatory elements. They further suggest that transcriptional regulation of Olig2 is involved in segregation of pMN neuroblasts. PMID- 16469307 TI - Calcium and magnesium in drinking water and risk of death from acute myocardial infarction in Taiwan. AB - Many studies have examined the association between cardiovascular disease mortality and water hardness. However, the results have not been consistent. This report examines whether calcium and magnesium in drinking water are protective against acute myocardial infarction (AMI). All eligible AMI deaths (10,094 cases) of Taiwan residents from 1994 to 2003 were compared with deaths from other causes (10,094 controls), and the levels of calcium and magnesium in drinking water of these residents were determined. Data on calcium and magnesium levels in drinking water throughout Taiwan have been obtained from the Taiwan Water Supply Corporation. The control group consisted of people who died from other causes and the controls were pair matched to the cases by sex, year of birth, and year of death. The adjusted odd ratios (95% confidence interval) were 0.79 (0.73-0.86) for the group with water calcium levels between 25.1 and 42.4 mg/L and 0.71 (0.65 0.77) for the group with calcium levels of 42.6 mg/L or more. After adjustment for calcium levels in drinking water, there was no difference between the groups with different levels of magnesium. The results of the present study show that there is a significant protective effect of calcium intake from drinking water on the risk of death from AMI. PMID- 16469308 TI - Kinase inhibitors and airway inflammation. AB - Kinases are believed to play a crucial role in the expression and activation of inflammatory mediators in the airway, in T-cell function and airway remodelling. Important kinases such as Inhibitor of kappaB kinase (IKK)2, mitogen activated protein (MAP) kinases and phsopho-inositol (PI)3 kinase regulate inflammation either through activation of pro-inflammatory transcription factors such as activating protein-1 (AP-1) and nuclear factor kappaB (NF-kappaB), which are activated in airway disease, or through regulation of mRNA half-life. Selective kinase inhibitors have been developed which reduce inflammation and some characteristics of disease in animal models. Targeting specific kinases that are overexpressed or over active in disease should allow for selective treatment of respiratory diseases. Interest in this area has intensified due to the success of the specific Abelson murine leukaemia viral oncogene (Abl) kinase inhibitor imatinib mesylate (Gleevec) in the treatment of chronic myelogenous leukaemia. Encouraging data from animal models and primary cells and early Phase I and II studies in other diseases suggest that inhibitors of p38 MAP kinase and IKK2 may prove to be useful novel therapies in the treatment of severe asthma, chronic obstructive pulmonary disease (COPD), cystic fibrosis and other inflammatory airway diseases. PMID- 16469309 TI - Antioxidant enzymes activity involvement in luteolin-induced human lung squamous carcinoma CH27 cell apoptosis. AB - Luteolin (3',4',5,7-tetrahydroxyflavone) is an active constituent of Lonicera japonica (Caprifoliaceae), and has been reported to produce anti-tumor activities. However, the apoptosis-inducing activity of luteolin still remains unknown. Flavonoids have been found to possess prooxidant and antioxidant action. The biological and pharmacological effect of flavonoid may depend upon its behavior as either an antioxidant or a prooxidant. Our experiments found that luteolin-induced CH27 cell apoptosis was accompanied by activation of antioxidant enzymes, such as superoxide dismutase and catalase, but not through the production of reactive oxygen species and disruption of mitochondrial membrane potential. Therefore, the effects of luteolin on CH27 cell apoptosis were suspected to result from the antioxidant rather than the prooxidant action of luteolin. PMID- 16469310 TI - Beta-adrenoceptor responses of the airways: for better or worse? AB - Beta2-adrenoceptor agonists are the first-line treatment of asthma and chronic obstructive pulmonary disease (COPD), in which a short-acting beta2-adrenoceptor agonist is used as required for relief of bronchoconstriction. A long-acting beta2-adrenoceptor agonist may be added to an inhaled corticosteroid as step 3 in the management of chronic asthma. Long-acting beta2-adrenoceptor agonists may also be added in treatment of COPD. This review examines the beneficial and detrimental effects of beta2-adrenoceptor agonists. The beneficial effects of beta2-adrenoceptor agonists are mainly derived from their bronchodilator activity which relieves the bronchiolar narrowing and improves air flow. The potential anti-inflammatory actions of stabilizing mast cell degranulation and release of inflammatory and bronchoconstrictor mediators, is considered. Other potential beneficial responses include improvements in mucociliary clearance and inhibition of extravasation of plasma proteins that is involved in oedema formation in asthma. The side effects of beta2-adrenoceptor agonists are primarily related to beta2-adrenoceptor-mediated responses at sites outside the airways. Of major concern has been the development of tolerance and this is discussed in relation to incidence of increased morbidity and mortality to asthma over the past three decades. A clinical aspect of beta2-adrenoceptor pharmacology in recent years has been the recognition of genetic polymorphism of the receptor and how this affects responses to and tolerance to beta2-adrenoceptor agonists. A controversial feature of beta2-adrenoceptor agonists is their stereoisomerism and whether the inactive (S)-isomer of salbutamol had detrimental actions in the commercially used racemate. The consensus is that despite these adverse properties, beta2 adrenoceptor agonist remains the most useful pharmacological agents in the management of asthma and COPD. PMID- 16469311 TI - SUMO modification through rapamycin-mediated heterodimerization reveals a dual role for Ubc9 in targeting RanGAP1 to nuclear pore complexes. AB - SUMOs (small ubiquitin-related modifiers) are eukaryotic proteins that are covalently conjugated to other proteins and thereby regulate a wide range of important cellular processes. The molecular mechanisms by which SUMO modification influences the functions of most target proteins and cellular processes, however, remain poorly defined. A major obstacle to investigating the effects of SUMO modification is the availability of a system for selectively inducing the modification or demodification of an individual protein. To address this problem, we have developed a procedure using the rapamycin heterodimerizer system. This procedure involves co-expression of rapamycin-binding domain fusion proteins of SUMO and candidate SUMO substrates in living cells. Treating cells with rapamycin induces a tight association between SUMO and a single SUMO substrate, thereby allowing specific downstream effects to be analyzed. Using RanGAP1 as a model SUMO substrate, the heterodimerizer system was used to investigate the molecular mechanism by which SUMO modification targets RanGAP1 from the cytoplasm to nuclear pore complexes (NPCs). Our results revealed a dual role for Ubc9 in targeting RanGAP1 to NPCs: In addition to conjugating SUMO-1 to RanGAP1, Ubc9 is also required to form a stable ternary complex with SUMO-1 modified RanGAP1 and Nup358. As illustrated by our studies, the rapamycin heterodimerizer system represents a novel tool for studying the molecular effects of SUMO modification. PMID- 16469312 TI - Refinement of the mouse model of congenital toxoplasmosis. AB - The goals of the present investigation, focusing on the BALB/c mouse model of congenital toxoplasmosis, were: (1) to find a method to determine pregnancy in the mouse. The method has 100% sensitivity and 72% specificity; (2) to test congenital transmission during the chronic stage of toxoplasmosis. This occurred in 2 of 10 mice tested; (3) to investigate the relationship between the infective dose and the rate of congenital transmission. This was not demonstrated for doses of 10(2) to 10(3) bradyzoites and oocysts of Prugniaud, M3 and M7741 strains, with transmission rates of 3 of 8 to 6 of 10 mice inoculated; (4) to determine homologous and heterologous protection. Homologous protection was demonstrated with Prugniaud cysts, and heterologous protection was found between ME-49 and M3 cysts. This finding is consistent with the uniform natural protection against congenital toxoplasmosis seen in immune women and ewes. PMID- 16469313 TI - Reconstitution of the enzyme AroA and its glyphosate tolerance by fragment complementation. AB - 5-Enolpyruvylshikimate-3-phosphate (EPSP) synthase (AroA) is a key enzyme in the aromatic amino acid biosynthetic pathway in microorganisms and plants, and is the target of the herbicide glyphosate. Glyphosate tolerance activity of the enzyme could be obtained by natural occurrence or by site-directed mutagenesis. A functional Pseudomonas putida AroA was obtained by co-expression of two protein fragments AroA(P. putida)-N210 and AroA(P. putida)-C212 in Escherichia coli aroA mutant strain AB2829. From sequence analysis, the equivalent split site on E. coli AroA was chosen for further study. The result indicated that functional E. coli AroA could also be reconstituted from two protein fragments AroA(E. coli) N218 and AroA(E. coli)-C219, under both in vivo and in vitro conditions. This result suggested that the fragment complementation property of this family of enzyme may be general. Additional experiments indicated that the glyphosate tolerance property of AroA could also be reconstituted in parallel with its enzyme activity. The implication of this finding is discussed. PMID- 16469314 TI - Antifungal activity of synthetic peptide derived from halocidin, antimicrobial peptide from the tunicate, Halocynthia aurantium. AB - Halocidin is an antimicrobial peptide isolated from the hemocytes of the tunicate. Among the several known synthetic halocidin analogues, di-K19Hc has been previously confirmed to have the most profound antibacterial activity against antibiotic-resistant bacteria. This peptide has been considered to be an effective candidate for the development of a new type of antibiotic. In this study, we have assessed the antifungal activity of di-K19Hc, against a panel of fungi including several strains of Aspergillus and Candida. As a result, we determined that the MICs of di-K19Hc against six Candida albicans and two Aspergillus species were below 4 and 16 microg/ml, respectively, thereby indicating that di-K19Hc may be appropriate for the treatment of several fungal diseases. We also conducted an investigation into di-K19Hc's mode of action against Candida albicans. Our colony count assay showed that di-K19Hc killed C. albicans within 30s. Di-K19Hc bound to the surface of C. albicans via a specific interaction with beta-1,3-glucan, which is one of fungal cell wall components. Di K19Hc also induced the formation of ion channels within the membrane of C. albicans, and eventually observed cell death, which was confirmed via measurements of the K+ released from C. albicans cells which had been treated with di-K19Hc, as well as by monitoring of the uptake of propidium iodide into the C. albicans cells. This membrane-attacking quality of di-K19Hc was also visualized via confocal laser and scanning electron microscopy. PMID- 16469316 TI - Functional and evolutionary analyses on expressed intronless genes in the mouse genome. AB - Using computational approaches we have identified 2017 expressed intronless genes in the mouse genome. Evolutionary analysis reveals that 56 intronless genes are conserved among the three domains of life--bacteria, archea and eukaryotes. These highly conserved intronless genes were found to be involved in essential housekeeping functions. About 80% of expressed mouse intronless genes have orthologs in eukaryotic genomes only, and thus are specific to eukaryotic organisms. 608 of these genes have intronless human orthologs and 302 of these orthologs have a match in OMIM database. Investigation into these mouse genes will be important in generating mouse models for understanding human diseases. PMID- 16469315 TI - Extracellular superoxide dismutase exists as an octamer. AB - Human extracellular superoxide dismutase (EC-SOD) is involved in the defence against oxidative stress induced by the superoxide radical. The protein is a homotetramer stabilised by hydrophobic interactions within the N-terminal region. During the purification of EC-SOD from human aorta, we noticed that material with high affinity for heparin-Sepharose formed not only a tetramer but also an octamer. Analysis of the thermodynamic stability of the octamer suggested that the C-terminal region is involved in formation of the quaternary structure. In addition, we show that the octamer is composed of both aEC-SOD and iEC-SOD folding variants. The presence of the EC-SOD octamer with high affinity may represent a way to influence the local concentration of EC-SOD to protect tissues specifically sensitive to oxidative damage. PMID- 16469317 TI - J1 acylase, a glutaryl-7-aminocephalosporanic acid acylase from Bacillus laterosporus J1, is a member of the alpha/beta-hydrolase fold superfamily. AB - J1 acylase, a glutaryl-7-aminocephalosporanic acid acylase (GCA) isolated from Bacillus laterosporus J1, has been conventionally grouped as the only member of class V GCA, although its amino acid sequence shares less than 10% identity with members of other classes of GCA. Instead, it shows higher sequence similarities with Rhodococcus sp. strain MB1 cocaine esterase (RhCocE) and Acetobacter turbidans alpha-amino acid ester hydrolase (AtAEH), members of the alpha/beta hydrolase fold superfamily. Homology modeling and secondary structure prediction indicate that the N-terminal region of J1 acylase has an alpha/beta-hydrolase folding pattern. The catalytic triads in RhCocE and AtAEH were identified in J1 acylase as S125, D264 and H309. Mutations to alanine at these positions were found to completely inactivate the enzyme. These results suggest that J1 acylase is a member of the alpha/beta-hydrolase fold superfamily with a serine-histidine aspartate catalytic triad. PMID- 16469318 TI - Low ambient temperature increases food intake and dropping production, leading to incorrect estimates of hormone metabolite concentrations in European stonechats. AB - Non-invasive methods to measure steroid hormone metabolites in bird droppings or mammalian feces have become very popular. However, the accuracy of these measurements may be affected by many factors. Here, we use the stonechat (Saxicola torquata) as a passerine bird model to test whether differences in ambient temperature affect food intake and dropping production and whether these changes lead to measurement artefacts in hormone metabolite concentrations. In addition, we tested for diurnal patterns in hormone metabolites. We held European stonechats in climate chambers and subjected them to two different long-term ambient temperature regimes, +5 degrees C and +22 degrees C. As expected, food intake and dropping production was higher at +5 degrees C than at +22 degrees C. Plasma concentrations of corticosterone and testosterone did not differ between different ambient temperature regimes. However, corticosterone and testosterone metabolite concentrations (in ng/g) were significantly lower at +5 degrees C than at +22 degrees C. When we measured the rate of hormone metabolite excretion (in picogram per hour) instead of the concentration, there was no difference between treatment groups. Thus, the measurement of hormone metabolite concentrations can be flawed because, depending on the treatment, similar amounts of hormone metabolites can be excreted into very different amounts of droppings. In conclusion, hormone metabolite concentration measurements are sensitive to changes in ambient temperature and probably any other factor that alters metabolic rates. Any study involving systematic changes in metabolism--i.e., during molt, migration, hibernation, egg production, or seasonal comparisons- needs to take these caveats into account. PMID- 16469319 TI - Intrauterine growth restriction. AB - This study reviewed the screening, diagnosis, prophylaxis, and treatment of intrauterine growth restriction using the PubMed database for key words and the Cochrane database for systematic reviews. Identification of risk factors and measurement of symphysis-fundus height are currently the screening standards. Diagnosis is verified by ultrasonography. Accuracy of diagnosis may be improved by using customized fetal growth curves, symphysis-fundus height charts, and 3 dimensional ultrasonographic evaluation and measuring umbilical artery Doppler dimensional ultrasonographic evaluation measuring umbilical artery Doppler impedance. Prophylaxis with acetylsalicylic acid, started in the first or second trimester or combined with heparin before conception, may reduce the incidence of growth restriction in specific groups at high risk. Active management may reduce incidence in patients with mild to moderate asthma, and targeted treatment of infections may also be beneficial. Antenatal corticosteroid treatment also reduces the perinatal morbidity and mortality associated with IUGR. Bed rest has no demonstrated beneficial effects. PMID- 16469320 TI - Characterisation and expression of an Hsp70 gene from Parastrongyloides trichosuri. AB - Parastrongyloides trichosuri is a nematode parasite of Australian brushtail possums that has an alternative free-living life cycle which can be readily maintained indefinitely in a laboratory setting. The ability to maintain this parasite in a free-living cycle and induce it to parasitism at the free-living L1 stage makes this an excellent model for the study of genes associated with parasitism. A 70kD protein from infective larvae of P. trichosuri that appears to be immunogenic in infected possums has been identified as a heat shock protein (Hsp)70 homologue. The complete gene for Pt-Hsp70 was cloned and sequenced. The protein encoded by the Pt-Hsp70 gene is the likely orthologue of the Caenorhabditis elegans protein, Hsp70A, also known as hsp-1. Reverse transcriptase-PCR data indicate that Pt-Hsp70 (designated Pt-hsp-1) is expressed at readily detectable levels in all developmental stages of both the parasitic and free-living P. trichosuri life cycles and the promoter is mildly inducible by heat shock. Bioinformatic analysis of expressed sequence tag databases indicates that C. eleganshsp-1 homologues, together with C. eleganshsp-3 homologues, are the predominant members of the Hsp70 superfamily that are normally expressed in parasitic stages of the Strongyloididae family. Promoter fusions to a beta galactosidase coding sequence were prepared and introduced into wild type C. elegans to produce transgenic nematodes. Reporter gene expression was clearly present within embryonic cells and within intestinal cells of larval and adult stages. Thus, the expression of the Pt-hsp-1 promoter within P. trichosuri and transgenic C. elegans appears similar to the known expression of C. elegans hsp 1. This promoter should be of value in efforts to develop genetic manipulation tools for P. trichosuri. PMID- 16469321 TI - Testing the efficacy of RNA interference in Haemonchus contortus. AB - RNA interference (RNAi) is widely used in Caenorhabditis elegans to identify gene function and has been adapted as a high throughput screening method to identify genes involved in essential processes. We have been examining whether RNAi could also be used on the strongylid parasitic nematode Haemonchus contortus to study gene function. Eleven genes were targeted in L1 and exsheathed L3 H. contortus larvae with RNAi methodologies which have been shown to be effective in C. elegans and parasitic nematodes-feeding, soaking and electroporation. Reverse transcriptase-PCR and, where possible, protein assays were carried out to examine decreases in mRNA and protein levels. RNAi soaking in dsRNA to beta-tubulin and sec-23, a gene involved in vesicle transport, resulted in specific decreases in mRNA levels in exsheathed L3 larvae. No signs of specific decreases in expression levels were observed for the other nine genes tested. Following electroporation of dsRNA in L1 stage larvae, significant decreases were observed for two out of four genes tested. These findings suggest that the RNAi pathway is functional in H. contortus and that, under certain conditions, it is possible to suppress gene expression by RNAi. However, it only works on a limited number of genes and in some cases the effect is small and difficult to reproduce. This indicates that the RNAi approaches established for C. elegans and other nematodes have limited efficacy in H. contortus. This may reflect differences between nematode species in dsRNA uptake and transport into cells and between cells. PMID- 16469322 TI - Mild folate deficiency induces a proatherosclerotic phenotype in endothelial cells. AB - Low folate/high homocysteine (Hcy) is an established risk marker for cardiovascular disease (CVD). Some in vivo studies suggest low folate may independently contribute to CVD. To study the effects of mild folate deficiency on endothelial function, we adapted the EA.hy 926 endothelial cell line to growth in medium containing 23 nM folic acid (LO cells) or 9 microM folic acid (HI cells). Folate derivatives were substantially depleted in LO cells relative to HI cells. No differences were seen in intracellular homocysteine, S adenosylmethionine (SAM), S-adenosylhomocysteine (SAH), the SAM:SAH ratio, or global DNA methylation, and there was no consistent difference in secreted homocysteine. A greater percentage of LO than HI cells were in S phase of the cell cycle; supplementation of LO cells with thymidine/hypoxanthine prevented this. LO cells were more elongated than HI cells and did not form tight monolayers. Stress fibers were very prominent in LO but not HI cells. Treatment of LO cells with rho kinase inhibitors abolished stress fibers and partially normalized cell shape. LO cell monolayers were more permeable than HI cell monolayers at confluence, and MCP-1 mRNA and protein expression was higher in LO than HI cells. Our results suggest that mild folate deficiency is proatherosclerotic. PMID- 16469323 TI - Different effects of oral conjugated estrogen and transdermal estradiol on arterial stiffness and vascular inflammatory markers in postmenopausal women. AB - AIMS: We compared the effects of oral conjugated equine estrogen (CEE) therapy and transdermal estradiol therapy on pulse wave velocity (PWV) and circulating levels of vascular inflammatory markers in postmenopausal women and we also explored the interrelationship between the change in PWV and the changes in vascular inflammatory markers. METHODS AND RESULTS: In a randomized 12-month trial, 28 postmenopausal women received a continuous oral CEE plus cyclic medroxyprogesterone acetate (MPA), 28 received a continuous transdermal estradiol patch plus cyclic MPA, and 27 did not receive either therapy. In each subject, we measured the brachial-ankle PWV (baPWV) using an automated device, the blood pressure, and the circulating levels of vascular inflammatory markers (C-reactive protein [CRP], cell adhesion molecules [CAMs], monocyte chemoattractant protein-1 [MCP-1], and matrix metalloproteinase [MMP-9]) before and 12 months after the start of the study. Oral CEE therapy did not change the baPWV but significantly increased the CRP and MMP-9 levels (P<0.05, each) and significantly decreased the CAMs and MCP-1 levels (P<0.05, each). Transdermal estradiol therapy significantly decreased the baPWV, and the CAMs and MCP-1 levels (P<0.05, each) but had no effect on the CRP or MMP-9 levels. No significant changes were seen in the control group. The blood pressures of the subjects remained unchanged. In the transdermal estradiol group, the change in baPWV was not significantly correlated with the changes in vascular inflammatory markers. CONCLUSION: Transdermal estradiol, but not oral CEE therapy, may have antiatherosclerotic effects by improving arterial stiffness. The reduction in baPWV may contribute to the direct effect of estrogen, but not to the decrease in estrogen-induced vascular inflammatory markers. PMID- 16469325 TI - Extended DLVO interactions between spherical particles and rough surfaces. AB - An "extended DLVO" approach that includes Lifshitz-van der Waals, Lewis acid base, and electrostatic double layer interactions is used to describe interaction energies between spherical particles and rough surfaces. Favorable, unfavorable, and intermediate deposition conditions are simulated using surface properties representing common aquatic colloids and polymeric membranes. The surface element integration (SEI) technique and Derjaguin's integration method are employed to calculate interaction energy. Numerical simulations using SEI demonstrate that nanometer scale surface roughness features can produce a distribution of interaction energy profiles. Local interaction energies are statistically analyzed to define representative interaction energy profiles-minimum, average, and maximum-for various combinations of simulated particles and surfaces. In all cases, the magnitude of the average interaction energy profile is reduced, but the reduction of energy depends on particle size, asperity size, and density of asperities. In some cases, a surface that is on average unfavorable for deposition (repulsive) may possess locally favorable (attractive) sites solely due to nanoscale surface roughness. A weighted average of the analytical sphere sphere and sphere-plate expressions of Derjaguin reasonably approximates the average interaction energy profiles predicted by the SEI model, where the weighting factor is based on the fraction of interactions involving asperities. PMID- 16469324 TI - Separation and on-line concentration of saponins from Panax notoginseng by micellar electrokinetic chromatography. AB - A simple, reproducible and sensitive micellar electrokinetic chromatography (MEKC) method was developed for the separation and determination of 10 saponins from Panax notoginseng. Field-enhanced sample injection with reverse migrating micelles (FESI-RMM) was used for on-line concentration of the saponins. The effects of concentrations of sodium dodecyl sulfate (SDS) and organic modifier, the sample matrix, the injection time of water plug, the injection voltage and injection time of sample on the separation and stacking efficiency were investigated. The optimum buffer contained 10mM H3PO4, 140 mM SDS, 20% acetonitrile and 15% 2-propanol and the pH of buffer was 2.4. The sample solution was diluted with 15 mM SDS and injected for 15s with -8 kV after injection of 2s water plug. Under the optimum conditions, the analytes were well separated with very high plate number (8.0 x 10(5)-1.2 x 10(6) N/m); the limits of detection of the analytes were 1.7-6.3 microg/mL. The high sensitivity permitted the determination of two minor saponins Rh1 (1.01 mg/g) and Rg2 (0.62 mg/g) in P. notoginseng, which were not determined in the literature. PMID- 16469326 TI - Thermodynamic quantities of surface formation of aqueous electrolyte solutions. VI. Comparison with typical nonelectrolytes, sucrose and glucose. AB - To demonstrate an important distinction between the electrolytes and nonelectrolytes, surface tension of aqueous solutions of typical nonelectrolytes, sucrose and glucose, was measured as a function of temperature and concentration. The presence of sucrose or glucose molecules in the surface region affects the surface tension in the same way as the presence of an ion does. There is, however, a difference in the temperature coefficient of the surface tension between typical nonelectrolyte solutions, sucrose and glucose, and alkali halide solutions. The entropy of surface formation of sucrose and glucose solutions is the same as that of pure water, while that of alkali halide solutions decreases with concentration. The relation between this entropy change and the formation of electric double layers was discussed. PMID- 16469327 TI - Solubility of ethylene in aqueous solution of sodium dodecyl sulfate at ambient temperature and near the hydrate formation region. AB - The solubility of ethylene in aqueous solutions of sodium dodecyl sulfate (SDS) at different concentrations was measured at temperature 298.2 K and near the hydrate formation region. The effect of SDS on the gas solubility was studied and the solubilities of ethylene in a single micelle under different conditions were evaluated. It was found that the micelle solubilization was obvious, especially in the region near hydrate formation conditions. The CMC of SDS solution was also evaluated based on the solubility vs SDS concentration curves and it was found that it decreased with decreasing temperature. PMID- 16469328 TI - Consonant and syllable structure patterns in childhood apraxia of speech: developmental change in three children. AB - Changes in consonant and syllable-level error patterns of three children diagnosed with childhood apraxia of speech (CAS) were investigated in a 3-year longitudinal study. Spontaneous speech samples were analyzed to assess the accuracy of consonants and syllables. Consonant accuracy was low overall, with most frequent errors on middle- and late-developing sounds. Omissions and substitutions were the dominant error types. Analysis of syllables revealed higher frequencies of error on complex mono- and polysyllables. Multiple regression analyses revealed that consonant accuracy is predicted by syllable shape accuracy and polysyllable frequency. Improvement was noted over time, although irregular patterns of consonant and syllable-level errors persisted across the period studied. Findings suggest that consonant errors in CAS are related to syllable-level deficits, namely difficulty constructing syllabic frames for speech production targets. LEARNING OUTCOMES: On the basis of this article, the reader will be able to (1) describe the deficits in consonant production demonstrated by the participants, (2) analyze the relationship between consonant production and syllable-level patterns of error and (3) consider the value of addressing syllable construction as a therapeutic goal. PMID- 16469329 TI - In vitro methods for generating CD8+ T-cell clones for immunotherapy from the naive repertoire. AB - Innovations in gene discovery and the analysis of gene expression are facilitating the identification of a growing number of antigens that could potentially be targeted for immunotherapy of tumors. Methods to reliably generate antigen-specific T-cell responses in vitro would be useful not only to screen candidate antigens for immunogenicity prior to embarking on in vivo vaccination trials, but also to generate T-cell lines or clones that could be used directly for adoptive immunotherapy approaches. Although many techniques have proven successful for expanding ex vivo effector cells from antigen-specific memory CD8(+) cells that have been primed in vivo, methods to reliably generate high avidity CTL clones from the naive repertoire have not been well described. Various methods for the induction and expansion of antigen-specific CD8(+) CTL clones from healthy A2(+) donors were compared, using WT1 as a model tumor associated antigen for which there is a low frequency of precursor T cells in naive individuals. In contrast to the well-studied Melan-A/MART-1 (Melan-A) A2 restricted response, for which the CD8(+) T-cell precursor frequency in the naive repertoire is unusually high, successful expansion of WT1-specific CD8(+) T cells appeared to be more dependent upon cell culture conditions. In particular, primary stimulation with autologous peptide-loaded monocyte-derived DC generated in 48 h (DC2d) was more effective in expanding WT1-reactive populations of CTL than stimulation with DC generated using the more standard week-long protocol (DC7d). Adding supplemental IL-7 2 to 3 days after initiation of a stimulation cycle expanded antigen-specific cells within CTL lines more efficiently than including the cytokine from the beginning of the cycle. Following primary stimulation with peptide-loaded mature DC, subsequent restimulation with peptide loaded PBMC as the stimulators was more effective at expanding antigen-specific cells than repeated stimulation with mature DC. Using these techniques, high avidity CTL clones specific for an A()0201-restricted epitope of WT1 have been generated from nearly all normal A2(+) donors tested. Such clones have been demonstrated to be capable of recognizing and lysing leukemic cells, and will soon be tested for therapeutic activity in clinical trials of adoptive immunotherapy in patients with relapsed leukemia after transplantation. PMID- 16469330 TI - Topological frustration and the folding of interleukin-1 beta. AB - The cytokine, interleukin-1beta (IL-1beta), adopts a beta-trefoil fold. It is known to be much slower folding than similarly sized proteins, despite having a low contact order. Proteins are sufficiently well designed that their folding is not dominated by local energetic traps. Therefore, protein models that encode only the folded structure and are energetically unfrustrated (Go-type), can capture the essentials of the folding routes. We investigate the folding thermodynamics of IL-1beta using such a model and molecular dynamics (MD) simulations. We develop an enhanced sampling technique (a modified multicanonical method) to overcome the sampling problem caused by the slow folding. We find that IL-1beta has a broad and high free energy barrier. In addition, the protein fold causes intermediate unfolding and refolding of some native contacts within the protein along the folding trajectory. This "backtracking" occurs around the barrier region. Complex folds like the beta-trefoil fold and functional loops like the beta-bulge of IL-1beta can make some of the configuration space unavailable to the protein and cause topological frustration. PMID- 16469331 TI - An atomic model of the thin filament in the relaxed and Ca2+-activated states. AB - Contraction of striated muscles is regulated by tropomyosin strands that run continuously along actin-containing thin filaments. Tropomyosin blocks myosin binding sites on actin in resting muscle and unblocks them during Ca2+ activation. This steric effect controls myosin-crossbridge cycling on actin that drives contraction. Troponin, bound to the thin filaments, couples Ca2+ concentration changes to the movement of tropomyosin. Ca2+-free troponin is thought to trap tropomyosin in the myosin-blocking position, while this constraint is released after Ca2+-binding. Although the location and movements of tropomyosin are well known, the structural organization of troponin on thin filaments is not. Its mechanism of action therefore remains uncertain. To determine the organization of troponin on the thin filament, we have constructed atomic models of low and high-Ca2+ states based on crystal structures of actin, tropomyosin and the "core domain" of troponin, and constrained by distances between filament components and by their location in electron microscopy (EM) reconstructions. Alternative models were also built where troponin was systematically repositioned or reoriented on actin. The accuracy of the different models was evaluated by determining how well they corresponded to EM images. While the initial low and high-Ca2+ models fitted the data precisely, the alternatives did not, suggesting that the starting models best represented the correct structures. Thin filament reconstructions were generated from the EM data using these starting models as references. In addition to showing the core domain of troponin, the reconstructions showed additional detail not present in the starting models. We attribute this to an extension of TnI linking the troponin core domain to actin at low (but not at high) Ca2+, thereby trapping tropomyosin in the OFF-state. The bulk of the core domain of troponin appears not to move significantly on actin, regardless of Ca2+ level. Our observations suggest a simple model for muscle regulation in which troponin affects the charge balance on actin and hence tropomyosin position. PMID- 16469332 TI - Nuclear transport of trimeric assembly intermediates exerts a morphogenetic control on the icosahedral parvovirus capsid. AB - The connection between nuclear transport and morphogenesis of a large macromolecular entity has been investigated using the karyophylic capsid of the parvovirus minute virus of mice (MVM) as a model. The VP1 (82 kDa) and VP2 (63 kDa) proteins forming the T = 1 icosahedral MVM capsid at the respective 1:5 molar ratio of synthesis, could be covalently cross-linked with dimethyl suberimidate into two types of oligomeric assemblies, which were present at stoichiometric amounts in infected cell extracts and purified viral particles. The larger species contained VP1 and corresponded in size (200 kDa) to a heterotrimer of one VP1 and two VP2 subunits. The smaller species contained VP2 only and corresponded in size (180 kDa) to a homotrimer. The introduction of bulky residues or the truncation of side-chains involved in multiple interactions at the interfaces between trimers of VPs in the MVM capsid, produced the accumulation of trimeric intermediates that were competent in nuclear translocation but not in capsid assembly. These results indicate that MVM maturation proceeds by cytoplasmic oligomerization of the capsid subunits into two types of trimers, which are the assembly intermediates competent to translocate across the nuclear membrane. Consistent with this conclusion, mutations at basic residues that inactivate a previously identified beta-stranded nuclear localization motif, which notably are not involved in inter or intra subunit contacts, led to cytoplasmic retention of the two types of trimers, with no evidence for other assembly intermediates. Although a fraction of the VP1 containing trimers were translocated into the nucleus driven by the conventional nuclear transport signal of VP1 N terminus, their further assembly in the absence of the VP2-only trimers yielded large molecular mass amorphous aggregates. Therefore, the nuclear transport stoichiometry of assembly intermediates may exert a morphogenetic quality control on macromolecular complexes like the MVM capsid. PMID- 16469333 TI - The structure of gelsolin bound to ATP. AB - Calcium activation of the actin-modifying properties of gelsolin is sensitive to ATP. Here, we show that soaking calcium-free gelsolin crystals in ATP-containing media results in ATP occupying a site that spans the two pseudosymmetrical halves of the protein. ATP binding involves numerous polar and hydrophobic contacts and is identical for the two copies of gelsolin related by non-crystallographic symmetry within the crystal. The gamma-phosphate of ATP participates in several charge-charge interactions consistent with the preference of gelsolin for ATP, as a binding partner, over ADP. In addition, disruption of the ATP-binding site through Ca2+ activation of gelsolin reveals why ATP binds more tightly to the inactive molecule, and suggests how the binding of ATP may modulate the sensitivity of gelsolin to calcium ions. Similarities between the ATP and PIP2 interactions with the C-terminal half of gelsolin are evident from their overlapping binding sites and in that both molecules bind more tightly in the absence of calcium ions. We propose a model for how PIP2 may bind to calcium-free gelsolin based on the ATP-binding site. PMID- 16469335 TI - Resource holding potential, subjective resource value, and game theoretical models of aggressiveness signalling. AB - Empirical evidence suggests that aggressiveness (willingness to enter into, or escalate an aggressive interaction) may be more important than the ability to win fights in some species. Both empirical and theoretical traditions treat aggressiveness as a distinct property from the ability (RHP) or motivation (subjective resource value) to win a fight. I examine how these three traits are clearly distinct when modelled using a simple strategic model of escalation. I then examine game theoretical models of agonistic communication and demonstrate that models in which aggressiveness is signalled require: (1) a trait, aggressiveness, which is neither a correlate, nor consequence of RHP or motivation, (2) a handicap which negates any benefit to be gained through the use of a particular signal, and (3) the absence of any other asymmetry which could be used to assign roles to players. I conclude that it is unlikely that these assumptions are ever met, and that empirical examples of "aggressiveness" are far more likely to represent long-term differences in subjective resource value. PMID- 16469334 TI - The effects of the checkpoints program on parent-imposed driving limits and crash outcomes among Connecticut novice teen drivers at 6-months post-licensure. AB - INTRODUCTION: Because crash rates are highly elevated during the first months of licensure, it is advisable for parents to limit teen driving so that teens can gain independent driving experience under less dangerous driving conditions. This report describes the effect of the Checkpoints Program on parent limits on novice teen driving through six months post-licensure. METHODS: Nearly one-quarter of all Connecticut teens who obtained a learner's permit over a 9-month period were recruited, providing a final sample of 3,743 who obtained licenses within the next 16 months. Families were randomized to the intervention or comparison condition. Intervention families received by mail a series of persuasive communications related to high-risk teen driving and a parent-teen driving agreement, while on the same schedule comparison families received standard information on driver safety. RESULTS: Families who participated in the Checkpoints Program reported significantly greater limits on teen driving at licensure, 3-months, and 6-months post-licensure. However, there were no differences in reported risky driving behavior, violations, or crashes. CONCLUSION: This is the first statewide study testing the efficacy of the Checkpoints Program. The results indicate that it is possible to foster modest increases in parental restrictions on teen driving limits during the first six months of licensure using passive persuasive communications, but that the levels of restriction obtained were not sufficient to protect against violations and crashes. PMID- 16469336 TI - An effective method to increase solvability in biochemical systems using S system. AB - In this paper we propose an effective method to estimate the intrinsic values in an immobilized enzyme system, i.e., Michaelis constant K(m) and the maximum reaction rate V(m). We combine three techniques: (1) the non-linear least square method for estimating the kinetic values, (2) orthogonal collocation with the Gauss integration method, and (3) Newton-Raphson method (NRM) or S-system method (SSM) as Newton-like method. We build a procedure to combine the first two methods to estimate the unknown kinetic values in a system. We apply this procedure to solve the intrinsic kinetic parameters determination problem in an immobilized enzyme systems following Michaelis-Menten reaction. To demonstrate the effectiveness of the current method, we test their convergence performance in detail. The results show that the basin of attraction in the current method is extremely enlarged compared with that of the S-system alone. We suggest that the current method is one of the most effective ways to solve fairly complicated biochemical reaction systems in general. PMID- 16469337 TI - Estimating kinetic parameters from HIV primary infection data through the eyes of three different mathematical models. AB - The dynamics of HIV-1 infection consist of three distinct phases starting with primary infection, then latency and finally AIDS or drug therapy. In this paper we model the dynamics of primary infection and the beginning of latency. We show that allowing for time delays in the model better predicts viral load data when compared to models with no time delays. We also find that our model of primary infection predicts the turnover rates for productively infected T cells and viral totals to be much longer than compared to data from patients receiving anti-viral drug therapy. Hence the dynamics of the infection can change dramatically from one stage to the next. However, we also show that with the data available the results are highly sensitive to the chosen model. We compare the results using analysis and Monte Carlo techniques for three different models and show how each predicts rather dramatic differences between the fitted parameters. We show, using a chi(2) test, that these differences between models are statistically significant and using a jackknifing method, we find the confidence intervals for the parameters. These differences in parameter estimations lead to widely varying conclusions about HIV pathogenesis. For instance, we find in our model with time delays the existence of a Hopf bifurcation that leads to sustained oscillations and that these oscillations could simulate the rapid turnover between viral strains and the appropriate CTL response necessary to control the virus, similar to that of a predator-prey type system. PMID- 16469339 TI - A new method for measuring reaction times for odour detection at iso-intensity: Comparison between an unpleasant and pleasant odour. AB - A psychophysical detection test was developed to measure the reaction time of human subjects to a pleasant and an unpleasant odour. The response latencies to stimulation with a malodour (valeric acid) and pleasant odorant (amyl acetate) were compared over a range of different stimulus strengths. By expressing reaction time as a function of detection rate, the responses to the two odours can be compared at iso-intensity across the concentration range. This is the first study that allows odorants to be compared at the same intensity over a range of concentrations. The malodour valeric acid was detected more rapidly than amyl acetate; at the 50% detection level the reaction time for the detection of amyl acetate was 1.74 s compared 1.36 s for valeric acid (380 ms or 22% faster). Women were significantly faster than men at detecting both the unpleasant (by 18%) and pleasant (by 26%) odour at the 50% detection level and this disparity increased with decreasing stimulus strength. In conclusion, we demonstrate the ability of a new method for the measurement of reaction times to odour detection to discriminate between two different odours - a malodour and a non-malodour. PMID- 16469338 TI - Chronic low-dose administration of nicotine facilitates recovery and synaptic change after focal ischemia in rats. AB - The current study examines the effects of chronic administration of nicotine on motor behavior after focal stroke in rats. Animals were trained in a tray reaching task for 2weeks and then they were divided into: (1) control+saline (2) control+nicotine (3) stroke+saline, and (4) stroke+nicotine groups. Lesions were produced by devascularization of the surface blood vessels of the sensorimotor cortex contralateral to the forepaw used for reaching. Forty-eight hours after the lesions, and for a total of 12days, animals received two daily injections of either nicotine (0.3mg/kg) or saline (0.9%). Animals were tested in a motor battery 1week after the lesions and every other week for a total of 7weeks. Pyramidal cells in forelimb and cingulate areas were then examined for dendritic length and branching using a Golgi-Cox procedure. Behavioral results demonstrated that by the end of the testing stroke+nicotine animals showed significant behavioral improvement relative to stroke+saline animals. Stroke+nicotine animals showed an increase in dendritic length and branching in pyramidal cells of the forelimb and cingulate areas. The results suggest that the behavioral enhancement in the stroke+nicotine group might be attributable to the enhanced dendritic growth in residual cortical motor regions. PMID- 16469340 TI - A selective serotonin reuptake inhibitor reduces REM sleep in the homing pigeon. AB - Avian and mammalian 'rapid eye movement' sleep (REM sleep) resemble each other in several aspects. However, the question of whether REM sleep has a shared evolutionary ancestry in birds and mammals has yet to be thoroughly explored. The brain regions and neurotransmitter systems involved in the generation of mammalian REM sleep are phylogenetically ancient, and are also found in extant birds and reptiles. Several pharmacological experiments in birds indicate that similar neural substrates are involved in the regulation of avian and mammalian sleep. However, because the drugs used in these studies generally resulted in non specific sleep loss, the neurochemical regulation of avian REM sleep in particular remains uncertain. The selective serotonin reuptake inhibitor (SSRI) zimelidine is known to reduce REM sleep in mammals. If avian REM sleep is similarly regulated by serotonin, it would be expected that an acute dose of a SSRI should also reduce avian REM sleep. To investigate a putative role of serotonin in the regulation of avian REM sleep, changes in sleep electroencephalogram (EEG) and behavior were recorded in five pigeons (Columba livia) after the administration of an acute dose of zimelidine. Our results demonstrate that the effects of zimelidine on avian REM sleep are comparable to those observed in mammals, indicating that serotonin may serve a similar function in the control of avian and mammalian REM sleep. PMID- 16469341 TI - Cytotoxic biotransformed products from cinobufagin by Mucor spinosus and Aspergillus Niger. AB - Cinobufagin (1) was one of important cardenolidal steroids and major components of Chan'Su, a famous traditional Chinese medicine. Bioconversion of cinobufagin by the fungi of Mucor spinosus and Aspergillus niger were investigated. Nine bioconversion products were obtained from M. spinosus and seven products from A. niger. Their structures were elucidated by high-resolution fast atom bombardment mass spectroscopy (HR-FAB-MS), extensive NMR techniques, including (1)H NMR, (13)C NMR, DEPT, (1)H-(1)H correlation spectroscopy (COSY), two-dimensional nuclear Overhauser effect correlation spectroscopy (NOESY), heteronuclear multiple quantum coherence (HMQC) and heteronuclear multiple bond coherence (HMBC). The in vitro cytotoxic activities against human hepatoma cells (HepG2, SMMC-7221 and BEL-7402) and human leukemia cells (K562, HL-60 and HEL) of all bioconversion products were determined by the MTT method, and their structure activity relationships (SAR) were discussed. PMID- 16469342 TI - Surprising evolutionary predictions from enhanced ecological realism. AB - A focus on the eco-evolutionary feedback continually operating between a population's evolution and its environment helps to appreciate the generality of ESS theory. Here we illustrate, through a sequence of four examples, how respecting such feedback in the evolutionary dynamics of quantitative traits may result in qualitatively unexpected outcomes. Reviewing existing insights and complementing these with new results, we show (1) that evolutionary matrix games are fundamentally degenerate and allow a natural unfolding, (2) that selection driven extinction may not be rare in nature, (3) that evolutionary epidemiology should not rely on R0 maximization, and (4) why the occurrence of Hardy-Weinberg proportions generically requires an evolutionary explanation. PMID- 16469343 TI - The evolutionary consequences of plasticity in host-pathogen interactions. AB - Interactions between individuals such as hosts and pathogens are often characterized by substantial phenotypic plasticity. Pathogens sometimes alter their exploitation strategies in response to defensive strategies adopted by their host and vice versa. Nevertheless, most game-theoretic models developed to explain the evolution of pathogen and host characteristics assume that no such plasticity occurs. Allowing for phenotypic plasticity in these models is difficult because one must focus on the evolution of pathogen and host reaction norms, and then allow for the potentially indefinite reciprocal changes in pathogen and host behaviour that occur during an infection as a result of their interacting reaction norms. Here, we begin to address these issues for a simple host-pathogen system in which the pathogen exhibits a level of virulence and the host exhibits a level of immune clearance. We find, quite generally, that plasticity promotes the evolution of higher levels of cooperation, in this case leading to reduced levels of both virulence and clearance. PMID- 16469344 TI - Replicative fidelity of lentiviral vectors produced by transient transfection. AB - Previous investigations have estimated the human immunodeficiency virus type 1 (HIV) base pair substitution rate to be approximately 10(-4) to 10(-5) per round of viral replication, and HIV has been hypothesized to be more error-prone than other retroviruses. Using a single cycle reversion assay, we unexpectedly found that the reversion rates of HIV, avian leukosis virus and Moloney murine leukemia virus were the same, within statistical error. Because both the viral enzyme reverse transcriptase (RT) and cellular RNA polymerase II (RNAP) are required for viral replication, we hypothesized that the similar reversion rates actually reflect the intrinsic error rate of RNAP, which is the enzyme common to all three retroviruses in the reversion assay. To address this possibility, HIV vectors with the U3 region replaced by a reporter reversion cassette were constructed and vector supernatant produced by transient transfection. All single integrant revertant cell lines showed the identical mutations at both long terminal repeats. This indicates that either RNAP or another cellular enzyme is responsible for these reversions, or that HIV RT only makes errors during first strand synthesis. Additionally, when HIV particles were rescued from an integrated vector as opposed to being produced by transient transfection, the reversion rate was significantly lower, suggesting that one or more factors in the virus-producing cells plays a role in the fidelity of retroviral replication. These results have implications regarding the fidelity of the transgene after transient transfection production of lentiviral vector supernatants. PMID- 16469345 TI - Molecular characterization of a novel simian immunodeficiency virus lineage (SIVtal) from northern talapoins (Miopithecus ogouensis). AB - Simian immunodeficiency viruses (SIVs) are found in an extensive number of African primates, and humans continue to be exposed to these viruses by hunting and handling of primate bushmeat and following occupational exposures to captive nonhuman primates. Here, we report the molecular characterization of a new SIV lineage, SIVtal, from wild-caught and captive talapoin monkeys (Miopithecus ogouensis) from Cameroon and U.S. zoos, respectively. Phylogenetic tree analyses of a small fragment in the pol gene indicated that all SIVtal strains clustered together forming a single species-specific lineage. Full-length sequence analysis for two strains, SIVtal-00CM266 and SIVtal-01CM8023, from wild-caught animals in Cameroon confirmed that SIVtal was distinct from all primate lentiviruses isolated so far and represents a new SIV lineage. Phylogenetic analyses in different viral genes showed a significant clustering of the SIVtal lineage with the Cercopithecus-specific SIVs. In addition, SIVtal and Cercopithecus-specific SIVs share functional motifs in Gag and Env that distinguish them from other primate lentiviruses. Like SIVsyk and SIVdeb, a vpu gene homologue was also absent in SIVtal. Although northern talapoins belong to the Miopithecus genus, their SIVs belong to the Cercopithecus SIV lineage, suggesting evolution from a common ancestor or cross-species transmission between both primate genera. PMID- 16469346 TI - The effect of genome length on ejection forces in bacteriophage lambda. AB - A variety of viruses tightly pack their genetic material into protein capsids that are barely large enough to enclose the genome. In particular, in bacteriophages, forces as high as 60 pN are encountered during packaging and ejection, produced by DNA bending elasticity and self-interactions. The high forces are believed to be important for the ejection process, though the extent of their involvement is not yet clear. As a result, there is a need for quantitative models and experiments that reveal the nature of the forces relevant to DNA ejection. Here, we report measurements of the ejection forces for two different mutants of bacteriophage lambda, lambdab221cI26 and lambdacI60, which differ in genome length by approximately 30%. As expected for a force-driven ejection mechanism, the osmotic pressure at which DNA release is completely inhibited varies with the genome length: we find inhibition pressures of 15 atm and 25 atm, for the short and long genomes, respectively, values that are in agreement with our theoretical calculations. PMID- 16469347 TI - Vaccination of ducks with a whole-cell vaccine expressing duck hepatitis B virus core antigen elicits antiviral immune responses that enable rapid resolution of de novo infection. AB - As a first step in developing immuno-therapeutic vaccines for patients with chronic hepatitis B virus infection, we examined the ability of a whole-cell vaccine, expressing the duck hepatitis B virus (DHBV) core antigen (DHBcAg), to target infected cells leading to the resolution of de novo DHBV infections. Three separate experiments were performed. In each experiment, ducks were vaccinated at 7 and 14 days of age with primary duck embryonic fibroblasts (PDEF) that had been transfected 48 h earlier with plasmid DNA expressing DHBcAg with and without the addition of anti-DHBcAg (anti-DHBc) antibodies. Control ducks were injected with either 0.7% NaCl or non-transfected PDEF. The ducks were then challenged at 18 days of age by intravenous inoculation with DHBV (5 x 10(8) viral genome equivalents). Liver biopsies obtained on day 4 post-challenge demonstrated that vaccination did not prevent infection of the liver as similar numbers of infected hepatocytes were detected in all vaccinated and control ducks. However, analysis of liver tissue obtained 9 or more days post-challenge revealed that 9 out of 11 of the PDEF-DHBcAg vaccinated ducks and 8 out of 11 ducks vaccinated with PDEF DHBcAg plus anti-DHBc antibodies had rapidly resolved the DHBV infection with clearance of infected cells. In contrast, 10 out of 11 of the control unvaccinated ducks developed chronic DHBV infection. In conclusion, vaccination of ducks with a whole-cell PDEF vaccine expressing DHBcAg elicited immune responses that induced a rapid resolution of DHBV infection. The results establish that chronic infection can be prevented via the vaccine-mediated induction of a core-antigen-specific immune response. PMID- 16469348 TI - Size-invariant but viewpoint-dependent representation of faces. AB - The present study investigated the role of size and view on face discrimination, using a novel set of synthetic face stimuli. Face discrimination thresholds were measured using a 2AFC match-to-sample paradigm, where faces were discriminated from a mean face. In Experiment 1, which assessed the effect of size alone, subjects had to match faces that differed in size up to four-fold. In Experiment 2 where only viewpoint was manipulated, a target face was presented at one of four different views (0 degree front, 6.7 degrees, 13.3 degrees, and 20 degrees side) and subsequent matches appeared either at the same or different view. Experiment 3 investigated how face view interacts with size changes, and subjects matched faces differing both in size and view. The results were as follows: (1) size changes up to four-fold had no effect on face discrimination; (2) threshold for matching different face views increased with angular difference from frontal view; (3) size differences across different views had no effect on face discrimination. Additionally, the present study found a perceptual boundary between 6.7 degrees and 13.3 degrees side views, grouping 0 degrees front and 6.7 degrees side views together and 13.3 degrees and 20 degrees side views together. This suggests categorical perception of face view. The present study concludes that face view and size are processed by parallel mechanisms. PMID- 16469349 TI - The long and the short of it: spatial statistics at fixation vary with saccade amplitude and task. AB - We recorded over 90,000 saccades while observers viewed a diverse collection of natural images and measured low level visual features at fixation. The features that discriminated between where observers fixated and where they did not varied considerably with task, and the length of the preceding saccade. Short saccades (<8 degrees) are image feature dependent, long are less so. For free viewing, short saccades target high frequency information, long saccades are scale invariant. When searching for luminance targets, saccades of all lengths are scale-invariant. We argue that models of saccade behaviour must account not only for task but also for saccade length and that long and short saccades are targeted differently. PMID- 16469350 TI - Flash lag in depth. AB - The perceived position of a moving target at a particular point in time, indicated by a flash, is often judged to be different from its actual location. Here, we show that the position of a target moving in depth is also systematically mislocalized. We used three types of targets moving in depth at a range of speeds from 2 to 16 cm/s. (i) A target realistically rendered that included concordant looming, disparity, and perspective cues. (ii) A random dot surface whose depth was defined by disparity, without concordant perspective or looming cues. (iii) A surface of dynamic random dots whose depth was defined by disparity with no consistent motion visible monocularly. Subjects viewed the targets moving either towards or away from them and indicated whether the targets appeared to be nearer or farther than a continuously present reference depth at the moment that a flash was presented. A staircase procedure was used to null, and thus measure, any perceptual displacement from the reference depth. A flash lag in depth was found in which the target appeared ahead of its true position, displaced by a constant amount of time depending on the stimulus type and the direction of motion (towards or away). The time displacement varied from 76 ms (for the realistic target moving away from the observer) to 263 ms (for static random dots moving towards). These effects may depend on the confidence with which subjects were able to judge the location of our various targets: greater confidence leading to a smaller temporal displacement. PMID- 16469351 TI - Multiple sources of information and time-to-contact judgments. AB - Four experiments examined how the visual system deals with multiple information sources for perceiving dynamic events. Two tau-type optical variables, one defined by the expanding object's image and the other defined by the expanding angular extent composed of the line of sight and the object's shadow, were manipulated in time-to-contact judgments. When the information specified by both variables was consistent, little perceptual accuracy was gained by having two information sources. When the two sources conflicted, perceptual accuracy deteriorated in proportion to the degree of conflict. Based on these results, we concluded that the visual system integrates multiple sources of event-specific information, and that a reliable source of information can be the shadows cast by moving objects. PMID- 16469352 TI - Metacontrast masking suggests interaction between visual pathways with different spatial and temporal properties. AB - We examined the spatiotemporal characteristics of metacontrast using sinusoidal grating stimuli as the target and mask for quantitative comparison with the functional properties of the visual cortex. The magnitude of metacontrast effects depended on the stimulus features such as the orientation and spatial frequency of the target and mask. The characteristics of metacontrast dynamically changed depending on the stimulus onset asynchrony (SOA). At short SOAs (0 to approximately 40 ms), metacontrast exhibited a high stimulus feature specificity and a low contrast sensitivity, whereas at long SOAs ( approximately 40 to 80 ms), metacontrast exhibited a low stimulus feature specificity and a high contrast sensitivity. We suggest that metacontrast is explained by the interaction between two parallel visual pathways: one with a low contrast sensitivity and a high feature specificity, and the other with a high contrast sensitivity and a low feature specificity. PMID- 16469353 TI - Illusory color mixing upon perceptual fading and filling-in does not result in 'forbidden colors'. AB - A retinally stabilized object readily undergoes perceptual fading. It is commonly believed that the color of the apparently vanished object is filled in with the color of the background because the features of the filled-in area are determined by features located outside the stabilized boundary. Crane, H. D., & Piantanida, T. P. (1983) (On seeing reddish green and yellowish blue. Science, 221, 1078 1080) reported that the colors that are perceived upon full or partial perceptual fading can be 'forbidden' in the sense that they violate color opponency theory. For example, they claimed that their subjects could perceive "reddish greens" and "yellowish blues." Here we use visual stimuli composed of spatially alternating stripes of two different colors to investigate the characteristics of color mixing during perceptual filling-in, and to determine whether 'forbidden colors' really occur. Our results show that (1) the filled-in color is not solely determined by the background color, but can be the mixture of the background and the foreground color; (2) apparent color mixing can occur even when the two colors are presented to different eyes, implying that color mixing during filling in is in part a cortical phenomenon; and (3) perceived colors are not 'forbidden colors' at all, but rather intermediate colors. PMID- 16469354 TI - Sinusoid = light bar + dark bar? AB - A sinusoidal grating can be viewed as a series of light and dark bars. Here we measure the contrast discrimination thresholds for light and dark bars individually, and find that the contrast discrimination thresholds for the whole sinusoid can be explained as ideal summation of the light and dark bar thresholds. We propose a model for light bar, dark bar, and sinusoidal contrast discrimination which involves local light adaptation and multiplicative noise. The model accounts for the data very well, and also accounts for contrast discrimination of light and dark edges. PMID- 16469355 TI - Stereo dynamics are not scale-dependent. AB - The experiments reported here focus on the temporal dynamics of stereopsis in an effort to shed light on how low level mechanisms might contribute to the execution of coarse-to-fine processing in the human stereo system. Because previous studies have used a variety of stimuli and configurations, we assess the effect of exposure duration on stereo thresholds using band-limited Gabor patches for a range of stimulus configurations. In preliminary studies, we found that the best stereo sensitivity-spatial frequency relationship was obtained when using configurations in which the size and target-reference spacing were consistent with spatially scaled stimuli. Sub-optimal stereo sensitivity as a function of spatial frequency was observed when the size and separation were fixed. Further, we found that the temporal properties of stereopsis were consistently sustained in nature irrespective of the stimulus spatial frequency content. This latter finding suggests that if coarse-to-fine stereo processing does occur it does not follow as a consequence of the dynamics of low-level disparity transduction. PMID- 16469356 TI - Enhancing biodegradability of priority substances (pesticides) by solar photo Fenton. AB - In this paper, we present the photo-Fenton treatment in a solar pilot-plant scale of several EU priority hazardous substances (Alachlor, Atrazine, Chlorfenvinphos, Diuron and Isoproturon) dissolved in water. The results have been evaluated not only from the point of view of contaminant disappearance and mineralisation, but also of toxicity reduction and enhancement of biodegradability. Degradation was monitored by total organic carbon, pesticide concentration by HPLC-UV, inorganics released by ion chromatography, and biodegradability by the Zahn-Wellens (Z-W) test. The total volume of the solar photoreactor, composed of compound parabolic collectors with a total area of 4.16m2, was between 70 and 82 L. The treatment was shown to be effective, mineralising all of the pesticides tested, both alone and in mixtures. In order to find out the conditions for biocompatibility using the photo-Fenton reaction as a pre-treatment step, wastewater inoculated with unacclimated municipal sludge containing pesticides after certain degradation time was evaluated by the Z-W test. Biodegradability was enhanced (70% considered biodegradable) by the photo-Fenton treatment after 12-25min. It may be concluded that the photo-Fenton treatment consistently enhances biodegradability of wastewater containing pesticides. PMID- 16469357 TI - The changing state of contamination in the Lagoon of Venice. Part 2: heavy metals. AB - In order to verify whether pollution is increasing or decreasing, in 25 locations uniformly distributed in the central part of the Lagoon of Venice, a transitional environment suffering from man's urban and industrial activities, the same sampling scheme was repeated three times (in 1987, 1993 and 1998) over a 12-year period during which the lagoonal environment underwent substantial changes. Superficial sediments were sampled and analysed for heavy metals and total organic carbon contents, grain size and density. In general heavy metal contents were found to be correlated, with concentrations above the background level, e.g., for Hg, a concentration factor of 24 was observed in 1987. A temporal decrease in concentrations was observed for most of the metals. Detailed analysis on a smaller spatial scale showed that contamination significantly decreases from the inner border of the lagoon seawards, as highlighted in contour maps. The role of the Porto Marghera industrial zone as a source of pollutants at the border of the Lagoon was confirmed. The decrease in contamination could not be attributed only to a decrease in the intensity of sources, but also to erosion processes, worsened by intensive harvesting of clams with hydraulic dredges. PMID- 16469358 TI - Diethylaminoethyl-cellulose clean-up of a large volume naphthenic acid extract. AB - The Athabasca oil sands of Alberta, Canada contain an estimated 174 billion barrels of bitumen. During oil sands refining processes, an extraction tailings mixture is produced that has been reported as toxic to aquatic organisms and is therefore collected in settling ponds on site. Investigation into the toxicity of these tailings pond waters has identified naphthenic acids (NAs) and their sodium salts as the major toxic components, and a multi-year study has been initiated to identify the principal toxic components within NA mixtures. Future toxicity studies require a large volume of a NA mixture, however, a well-defined bulk extraction technique is not available. This study investigated the use of a weak anion exchanger, diethylaminoethyl-cellulose (DEAE-cellulose), to remove humic like material present after collecting the organic acid fraction of oil sands tailings pond water. The NA extraction and clean-up procedure proved to be a fast and efficient method to process large volumes of tailings pond water, providing an extraction efficiency of 41.2%. The resulting concentrated NA solution had a composition that differed somewhat from oil sands fresh tailings, with a reduction in the abundance of lower molecular weight NAs being the most significant difference. This reduction was mainly due to the initial acidification of tailings pond water. The DEAE-cellulose treatment had only a minor effect on the NA concentration, no noticeable effect on the NA fingerprint, and no significant effect on the mixture toxicity towards Vibrio fischeri. PMID- 16469359 TI - Photolytic degradation of quinalphos in natural waters and on soil matrices under simulated solar irradiation. AB - The photochemical persistence of quinalphos, one of the most widely used organophosphorous insecticides, was investigated in a variety of environmental matrices such as natural waters and soils of different composition. Simulated solar irradiation was obtained using a xenon arc lamp (Suntest CPS+ apparatus) giving an irradiation intensity of 750 W m(-2) equivalent to a light dose per hour of irradiation of 2,700 kJ m(-2). The phototransformation rates were determined using solid-phase microextraction (SPME) and ultrasonic extraction (USE) coupled to GC-FTD, while the identification of photoproducts was carried out by GC-MS. In water samples, the degradation kinetics followed a pseudo-first order reaction and photolysis half-lives ranged between 11.6 and 19.0 h depending on the constitution of the irradiated media. Dissolved organic matter (DOM) has a predominant retarding effect, while nitrate ions accelerated the photodegradation kinetics. In soil samples, the degradation kinetics was monitored on 1mm soil layer prepared on glass TLC plates. The kinetic behaviour of quinalphos was complex and characterized by a double step photoreaction, fast in the first 4h of irradiation followed by a slow degradation rate up to 64 h. The photolysis half life of quinalphos was shorter in sandy soil compared to the rest of the soil samples, varying between 16.9 and 47.5 h, and showing a strong dependence on the composition of the irradiated media. Among the transformation products formed mainly through photohydrolysis and photoisomerization processes, some photoproduct structures were proposed according to their mass spectral information. PMID- 16469360 TI - Biodegradation of 2,4,6-trichlorophenol and associated hydraulic conductivity reduction in sand-bed columns. AB - The aim of this research was to investigate the long-term hydraulic conductivity changes in sand-bed columns exposed to 2,4,6-trichlorophenol (TCP). Continuous flow laboratory studies were conducted using sand-bed columns (15 cm i.d.; 200 cm length) at 20+/-1 degrees C during 365 d. The influence of (i) initial loads of 2,4,6-TCP (15, 30, 45 and 60 mg kg(-1) of 2,4,6-TCP), and (ii) recirculating water velocity (0.09, 0.56 and 1.18 cm min(-1)) on the biodegradation of 2,4,6 TCP and hydraulic conductivity changes in the sand-bed columns were investigated. The experimental results indicated that biodegradation of 2,4,6-TCP followed pseudo-first-order kinetics in the range of k(1)=0.01-1.64 d(-1), and it was influenced by initial load (p<0.01) and recirculating water velocity (p<0.01). Indigenous microbial biomass growth and changes resulted in a spatial (180 cm) and temporal (365 d) reduction of hydraulic conductivity in the sand-bed columns by up to two orders of magnitude during biodegradation of 2,4,6-TCP. The fastest hydraulic conductivity reductions were observed in the sand-bed column operated at the highest recirculating water velocity and highest cumulative load of 2,4,6 TCP following 365 d of continuous treatment (p<0.05). PMID- 16469361 TI - Raw material use and behavioral modernity: Howiesons Poort lithic foraging strategies. PMID- 16469362 TI - The lack of association between four point mutations in the promoter region of the toll-like 4 receptor gene and myocardial infarction. AB - The toll-like receptor 4 gene product (TLR4) has been implicated in the pathogen recognition response mechanism because it plays a central role in the transcriptional activation of the host defense system. Activation of TLR4 initiates an intracellular signaling cascade, via the adapter protein MyD88 (myeloid differentiation factor 88), which leads to the activation of NF-kappaB transcriptional factor, and ultimately to the induction of a pro-inflammatory response. This inflammatory response has been increasingly associated with atherosclerosis. Recent analyses on two polymorphisms of TLR4, which affect the extracellular domain of the receptor, have been shown to give rise to an attenuated innate immune defense which may contribute to disease susceptibility. We have investigated the significance of four new substitutions found by re sequencing in the 5'-proximal promoter region of the TLR4 gene in a case-control study of acute myocardial infarction. Our results found no statistically significant association between these genetic variants and MI. PMID- 16469363 TI - Aqueous extracts of Ocimum basilicum L. (sweet basil) decrease platelet aggregation induced by ADP and thrombin in vitro and rats arterio--venous shunt thrombosis in vivo. AB - OBJECTIVE: To study the effects of aqueous extract of Ocimum basilicum L (OBL) on platelet aggregation and experimental thrombus. METHODS: Platelet aggregation induced by ADP (5 muM) and thrombin (4 UI), and thrombus weight in an arteriovenous thrombosis (AVT) model were tested after 2 weeks treatment with 15, 75 and 375 mg/kg OBL orally in rats, compared to 8.8 mg/kg/day aspirin. AVT was also tested 2 h after 75 mg/kg OBL orally, after 3 and 7 days treatment, and one, three and seven days after the end of a two-week treatment. Analysis was done by ANOVA followed by protected t-tests (Tukey). RESULTS: OBL (15, 75, 375 mg/Kg) dose-dependently inhibits platelet aggregation by ADP and thrombin, with 75 mg/kg/day having approximately the same effect as 8.8 mg/kg/day aspirin. ADP induced aggregation reached 45%, 28% and 18% for OBL, respectively, 15, 75, 375 mg/kg compared to 71% for control and 27% for aspirin (all p<0.01 except aspirin vs. OBL 75 mg/kg/day p=0.7). Thrombin-induced aggregation reached 33%, 22%, 21% for OBL, respectively, 15, 75, 375 mg/kg compared to 67% for control and 48% for aspirin (all p<0.01 except OBL 75 vs. OBL 375 mg/kg/day, p=1.0). Compared to a control thrombus weight of 48.1 mg (SD 4.9), thrombus weight was 29.4 (3.3), 19.0 (1.9) and 12.3 (1.7) after treatment for 2 weeks with 15, 75 and 375 mg/kg OBL, respectively, and 27.4 (5.3) after 8.8 mg/kg aspirin (all p<0.001 except aspirin vs. OBL 75 mg/kg/day p=1.0). Maximum effect of OBL was reached after one week's treatment. The effect subsided between 3 and 7 days. CONCLUSION: OBL possesses an inhibitory effect on platelet aggregation induced by ADP and thrombin, that is dose-dependent and results in an anti-thrombotic effect in vivo which develops progressively over 7 days and disappears over 3-7 days. The active ingredient now needs to be characterized. PMID- 16469364 TI - The prevalence of the prothrombin gene variant C20209T in African-Americans and Caucasians and lack of association with venous thromboembolism. PMID- 16469365 TI - MR imaging in endometrial carcinoma as a diagnostic tool for the absence of myometrial invasion. AB - OBJECTIVE: To evaluate whether magnetic resonance imaging (MRI) is a reliable modality for verifying preoperative diagnosis of stage IA endometrial carcinoma. METHODS: One hundred sixteen patients with endometrial carcinoma underwent preoperative pelvic non-contrast T2-weighted or dynamic MRI. We compared the interpretations of the MRI results with the histological findings of the resected uterus. RESULTS: In assessing the depth of myometrial invasion, the accuracy of MRI was 62.1%. As to the presence of cancerous myometrial invasion, the positive predictive value was 94.4% as high as previously reported by other institutions. However, the negative predictive value, the probability of the absence of myometrial invasion, was only 42.2%. Even when dynamic study was applied to the patient, the value only improved up to 60.0%. CONCLUSION: MRI has a definite advantage in evaluating deep myometrial invasion, but not the absence of invasion. We should take precautions against the risk of under-diagnosis when selecting stage IA endometrial carcinoma with use of MRI to preserve fertility or to eliminate lymphadenectomy. PMID- 16469366 TI - Effectiveness of creatine monohydrate on seizures and oxidative damage induced by methylmalonate. AB - Methylmalonic acidemias are metabolic disorders caused by a severe deficiency of methylmalonyl CoA mutase activity, which are characterized by neurological dysfunction, including convulsions. It has been reported that methylmalonic acid (MMA) accumulation inhibits succinate dehydrogenase (SDH) and beta hydroxybutyrate dehydrogenase activity and respiratory chain complexes in vitro, leading to decreased CO2 production, O2 consumption and increased lactate production. Acute intrastriatal administration of MMA also induces convulsions and reactive species production. Though creatine has been reported to decrease MMA-induced convulsions and lactate production, it is not known whether it also protects against MMA-induced oxidative damage. In the present study we investigated the effects of creatine (1.2-12 mg/kg, i.p.) and MK-801 (3 nmol/striatum) on the convulsions, striatal content of thiobarbituric acid reactive substances (TBARS) and on protein carbonylation induced by MMA. Moreover, we investigated the effect of creatine (12 mg/kg, i.p.) on the MMA induced striatal creatine and phosphocreatine depletion. Low doses of creatine (1.2 and 3.6 mg/kg) protected against MMA-induced oxidative damage, but did not protect against MMA-induced convulsions. A high dose of creatine (12 mg/kg, i.p.) and MK-801 (3 nmol/striatum) protected against MMA-induced seizures (evidenced by electrographic recording), protein carbonylation and TBARS production ex vivo. Furthermore, acute creatine administration increased the striatal creatine and phosphocreatine content and protected against MMA-induced creatine and phosphocreatine depletion. Our results suggest that an increase of the striatal high-energy phosphates elicited by creatine protects not only against MMA-induced convulsions, but also against MMA-induced oxidative damage. Therefore, since NMDA antagonists are limited value in the clinics, the present results indicate that creatine may be useful as an adjuvant therapy for methylmalonic acidemic patients. PMID- 16469367 TI - GnRH-agonist induction of fertile estrus with either natural mating or artificial insemination, followed by birth of pups in gray wolves (Canis lupus). AB - Although captive populations of endangered species such as the Mexican gray wolf (Canis lupus baileyi) can benefit from artificial insemination to accomplish genetic exchange, reliable techniques for timing insemination are lacking. We used the generic gray wolf (C. lupus) to test the efficacy of a short-acting GnRH agonist implant, deslorelin, for inducing estrus. Of five females receiving implants on 17 or 18 January 2003, two mated naturally 10-17 days later, and the others were artificially inseminated using fresh semen, one on day 7 and all three on day 11. Relaxin tests revealed that one artificially inseminated female and both naturally mated females were pregnant on 1 March, and all three gave birth to healthy puppies on 4-6 April. Of the artificially inseminated females, only the one who subsequently conceived and gave birth was judged to be in cytologic estrus at the time of insemination. Two females were treated again with deslorelin on 12 January 2004, followed by collection of fecal samples for hormone analysis. One female, who was housed with a male, copulated on day 17 but did not conceive; the other was not with an adult male. Fecal progestin and estrogen profiles suggested that estrus, but not ovulation, was induced. These results indicated that deslorelin could induce fertile estrus in the gray wolf, although individual response varied. Further investigation is needed to better define and control the interval between implant insertion and ovulation for optimal timing of insemination. PMID- 16469368 TI - In vitro canine oocyte nuclear maturation in homologous oviductal cell co-culture with hormone-supplemented media. AB - The aim of the present research was to verify the influence of oviductal cell co culture previously supplemented with steroids (estrogen, progesterone, or both) on IVM rates for oocytes from anestrous bitches that were cultured in vitro for 48, 72 and 96 h. Oocytes harvested from anestrous bitches were selected and allocated into four groups: Group 1 (co-culture in oviductal epithelial cells without hormonal supplementation-control); Group 2 (estrogen supplementation); Group 3 (progesterone supplementation); Group 4 (estrogen+progesterone supplementation). The oviductal epithelial cell culture was established 72 h prior to oocyte co-culture. After periods of 48, 72 and 96 h, the degree of oocyte nuclear maturation was assessed. Co-culture in oviductal epithelial cells with estrogen was not as beneficial for canine IVM as supplementation with progesterone and estrogen, or progesterone supplementation alone. Therefore, it was feasible to use co-culture with oviductal epithelial cells obtained from anestrous bitches for IVM (monolayer culture with oviduct cells previously supplemented with progesterone). Final stages of oocyte maturation were achieved at 72 and 96 h of culture; therefore, the duration of maturation for oocytes obtained from bitches in different stages of the estrous cycle should be taken into account. PMID- 16469369 TI - In vitro PGF2alpha production by endometrium and corpus luteum explants from pregnant and nonpregnant diestrus bitches and placental explants from pregnant bitches. AB - To better understand the process of slow luteal regression of the nonpregnant cycle in dogs and the acute luteolysis that occurs prepartum, the present study investigated in vitro PGF2alpha production by the endometrium, corpus luteum and placental explants obtained at known times of the cycle from pregnant bitches (days 63, 64 and immediately postpartum; day 0 = estimated day of the ovulatory LH surge) and from nonpregnant diestrus bitches (approximately days 65, 75 and 85). Both basal PGF2alpha production and its production in the presence of the protein kinase C (PKC) stimulator 12,13-phorbol dibutyrate (PDBu) were determined. For PDBu-supplemented incubations, mean PGF2alpha production (pg/mL/mg/6 h) by endometrium explants of the nonpregnant bitches in late diestrus was highest on day 65 (205 +/- 87) and reduced to low levels (38 +/- 17 and 11 +/- 11) on days 75 and 85, respectively. The production by corpus luteum explants from these bitches was significantly less on day 65 (46 +/- 14) than that of the day 65 endometrium explants, and was slightly increased on day 85 (103 +/- 52). The corresponding mean PGF2alpha production by the endometrium explants of pregnant bitches was on average much greater (i.e., two to three fold) compared to nonpregnant bitches (P < 0.01) and involved high concentrations at day 64 (1523 +/- 467) and postpartum, compared to somewhat lower levels on day 63 (830+/-65); luteal PGF production (165 +/- 4) was also higher than in nonpregnant bitches around day 65. For pregnant bitches, PGF production per gram of tissue in the endometrium explants was greater than for the CL or placenta explants (180 +/- 37). Therefore, the endometrium of the pregnant bitch has an increased capability to produce PGF2alpha immediately prepartum, which on a tissue weight basis, exceeds that of either corpora lutea or the placenta. However, assuming a larger mass of placental tissue in vivo, we inferred that the placenta may contribute substantially to peripheral PGF concentrations. PMID- 16469370 TI - Artificial insemination with frozen semen in dogs: a retrospective study of 10 years using a non-surgical approach. AB - From 1994 to 2003, a total of 526 bitches of 99 different breeds were artificially inseminated in 685 estrus cycles with domestic (n = 353) or imported (n = 332) frozen-thawed semen from 368 males. The overall whelping rate was 73.1% and mean (+/- S.E.M.) litter size 5.7 +/- 0.1 pups. The whelping rate was higher after intrauterine insemination (75.0%; n = 665) than after intravaginal insemination (10.0%, n = 20; P < 0.05). Insemination at the optimal time resulted in a higher whelping rate (78%, n = 559; P < 0.01) and larger litter size (5.8 +/ 0.2; P < 0.05) than inseminations performed late or too late (55.7% and 4.5 +/- 0.5, n = 61). Two inseminations (n = 384) yielded a higher whelping rate (P < 0.05) and mean litter size (P < 0.01) than one insemination (n = 241), 78.1% and 6.0 +/- 0.2 and 70.5% and 5.1 +/- 0.2, respectively. For inseminations performed at the optimal time, however, the whelping rate was not significantly different for bitches inseminated twice (79.3%, n = 358) versus once (76.8%, n = 168), but the litter size was larger (6.0 +/- 0.2 and 5.3 +/- 0.3). Semen classified as of poor quality (progressive motility < 50% or percentage abnormal sperm > 20%) resulted in a lower whelping rate (P < 0.01) than semen classified as of good quality (progressive motility > or = 50% and percentage abnormal sperm < or = 20%), 61 and 77%, respectively. Small breeds (n = 50) had a smaller litter size (3.9 +/- 0.3; P < 0.01) than larger breeds (medium [5.7 +/- 0.3, n = 94], large [5.9 +/- 0.2, n = 295] or giant breeds [6.1 +/- 0.5, n = 62] [P < 0.01]). Bitches older than 6 years had a lower whelping rate (68.2%) than younger ones (77.0%; P < 0.05). The duration of pregnancy was longer (P < 0.01) for bitches with a litter size of < 3 pups (61.7 +/- 0. 4 days, n = 30) than for bitches with larger litters (60.5 +/- 0.1 days, n = 177). These results show the potential of transcervical intrauterine insemination for routine artificial insemination in dogs. The results with frozen semen inseminations were optimised by inseminating bitches < or = 6 years old 2 and 3 days after ovulation with semen of good quality from males < or = 8 years old. PMID- 16469371 TI - RNA integrity and the effect on the real-time qRT-PCR performance. AB - The assessment of RNA integrity is a critical first step in obtaining meaningful gene expression data. Working with low-quality RNA may strongly compromise the experimental results of downstream applications which are often labour-intensive, time-consuming, and highly expensive. Using intact RNA is a key element for the successful application of modern molecular biological methods, like qRT-PCR or micro-array analysis. To verify RNA quality nowadays commercially available automated capillary-electrophoresis systems are available which are on the way to become the standard in RNA quality assessment. Profiles generated yield information on RNA concentration, allow a visual inspection of RNA integrity, and generate approximated ratios between the mass of ribosomal sub-units. In this review, the importance of RNA quality for the qRT-PCR was analyzed by determining the RNA quality of different bovine tissues and cell culture. Independent analysis systems are described and compared (OD measurement, NanoDrop, Bioanalyzer 2100 and Experion). Advantage and disadvantages of RNA quantity and quality assessment are shown in performed applications of various tissues and cell cultures. Further the comparison and correlation between the total RNA integrity on PCR performance as well as on PCR efficiency is described. On the basis of the derived results we can argue that qRT-PCR performance is affected by the RNA integrity and PCR efficiency in general is not affected by the RNA integrity. We can recommend a RIN higher than five as good total RNA quality and higher than eight as perfect total RNA for downstream application. PMID- 16469372 TI - An in vivo evaluation of bonding ability of comprehensive antibacterial adhesive system incorporating MDPB. AB - OBJECTIVES: This study examined the in vivo bonding ability to sound dentin of antibacterial adhesive systems incorporating an antibacterial monomer MDPB based on morphological evaluation of the resin-dentin interface. METHODS: Class V cavities were prepared on the buccal surfaces of the teeth of a beagle dog and a composite filling performed using (1) commercial self-etching system Liner Bond 2 (LB primer+LB bond), (2) experimental primer containing 5% MDPB and LB bond, (3) LB primer and experimental bonding-resin containing 2.5% MDPB, or (4) combination of experimental primer and bonding-resin. After 7 days, the tooth crown was cut and fixed in half-Karnovsky's solution, and the sectioned surface observed under scanning electron microscopy (SEM) after treatment with phosphoric acid and NaOCl. The ultrastructure of the bonding interface was also examined by transmission electron microscopy (TEM). Microtensile bond strengths (microTBS) of each group were measured using extracted teeth. RESULTS: SEM demonstrated that all groups produced a 1-2microm thick hybrid layer with funnel shaped resin tags, although the length of tags was shorter for the group in which MDPB-containing bonding-resin was used. TEM examination supported good adhesion of the comprehensive adhesive system employing MDPB-containing primer/bonding-resin, showing integrity between resin and dentin. There were no significant differences in microTBS among the four groups tested (p>0.05, ANOVA). SIGNIFICANCE: This study confirmed that the experimental antibacterial adhesive systems employing MDPB-containing primer or/and bonding-resin could produce an effective bond under in vivo conditions. PMID- 16469373 TI - Variation of depth of cure and intensity with distance using LED curing lights. AB - OBJECTIVES: The purpose of the study was to determine the correlation between intensity of light-emitting diode (LED) and tungsten-halogen light sources, and depth of cure of a resin composite at different distances. METHODS: Four LED curing lights (Flashlite 1001, Freelight 2, Smartlite IQ and Ultralume 5) and one tungsten halogen (Optilux 501, with 8 and 11 mm tips) were evaluated. Intensity was measured according a modified ISO Standard 10650 at distances of 0, 2, 4, 6, 8, 10 mm between the light tip and detector. Depth of cure (DOC) of TPH Spectrum shade A2 was measured according to the international standard ISO 4049 at the same distances. RESULTS: For all lights, intensity decreased as distance increased. The authors documented a logarithmic correlation between intensity and distance for all lights except the Smartlite IQ, Ultralume 5 and the Optilux 501 with the 11 mm tip, which showed a linear relationship between intensity and distance. All lights demonstrated a logarithmic correlation between intensity and DOC, and a linear correlation between DOC and distance. Smartlite IQ and Optilux 501 (11 mm tip) also had the least reduction in intensity and DOC at 10 mm. SIGNIFICANCE: Clinicians often an experience difficulty placing the light tip close to the resin surface when curing resin composites. While both intensity and DOC decrease with increasing distance, the relationship between these factors and distance may not be similar for all lights and may depend on the characteristics of individual lights. PMID- 16469374 TI - Solution properties of targacanthin (water-soluble part of gum tragacanth exudate from Astragalus gossypinus). AB - Solution properties of tragacanthin (the water-soluble part of gum tragacanth) were studied by gel permeation chromatography (GPC) combined with multi-angle light scattering and viscometry at 25 degrees C. Photon correlation spectroscopy was used to determine the hydrodynamic radius. Ultrasonic degradation was applied to obtain biopolymer fractions of different molecular weights. The dependence of intrinsic viscosity [eta] and radius of gyration (s2)z(1/2) on weight average molecular mass M(w) for this biopolymer were found to be [eta] = 9.077 x 10(-5) M(w)(0.87) (dL g(-1)) and (s2)z(1/2) in the range of M(w) from 1.8 x 10(5) to 1.6 x 10(6). The conformational parameters of tragacanthin were calculated to be 1111 nm for molar mass per unit contour length (M(L)), 26 nm for persistence length (q) and 1.87 ratio of R(g)/R(h). It was found that the Smidsrod parameter B, the empirical stiffness parameter was 0.013, which is lower than that of several polysaccharides indicating the stiff backbone for tragacanthin. The rheological behavior of aqueous solutions of gum tragacanth and its insoluble and soluble fractions (bassorin and tragacanthin, respectively) were studied. For concentrations equal to 1%, at 25 degrees C and in the absence of salt, bassorin solution showed the highest viscosity and shear thinning behaviour. Power law and Williamson models were used to describe the rheological behaviour of bassorin and tragacanthin, respectively. Oscillatory shear experiments showed a gel like structure for the bassorin but for tragacanthin the oscillatory data were as would be expected for semi-dilute to concentrated solution of entangled, random coil polymers. NaCl changed the steady and oscillatory rheological properties of both fractions and in this way the final viscosity of bassorin was even less than tragacanthin. The calculated activation energy for bassorin and tragacanthin indicated a more rapid decrease in viscosity with temperature for tragacanthin. The plot of eta(sp,0) versus C[eta] revealed that the transition from dilute to semi-dilute regime occurs at C*[eta] = 2.82 for tragacanthin. PMID- 16469375 TI - Effective transfection of cells with multi-shell calcium phosphate-DNA nanoparticles. AB - Coated calcium phosphate nanoparticles were prepared for cell transfection. A calcium phosphate nanoparticle served as core which was then coated with DNA for colloidal stabilisation. The efficiency of transfection could be considerably increased by adding another layer of calcium phosphate on the surface, thereby incorporating DNA into the particle and preventing its degradation within the cell by lysosomes. A subsequent outermost layer of DNA on the calcium phosphate gave a colloidal stabilisation. The efficiency of such multi-shell particles was significantly higher than that of simple DNA-coated calcium phosphate nanoparticles. The transfection efficiency of EGFP-encoding DNA was tested with different cell lines (T-HUVEC, HeLa, and LTK). The dispersions were stable and could be used for transfection after 2 weeks of storage at 4 degrees C without loss of efficiency. PMID- 16469376 TI - Placental oxidative stress in a rat model of preeclampsia. AB - The onset of preeclampsia is associated with increased maternal insult that could affect placental function. By increasing sodium intake (0.9% or 1.8% NaCl in drinking water) during the last week of gestation in the rat, we developed an animal model that shows many characteristics of preeclampsia such as increased blood pressure, decreased circulatory volume and diminished activity of the renin angiotensin-aldosterone system. The aim of the present study was to determine in this model whether maternal perturbations in pregnancy lead to placental oxidative stress. Sprague-Dawley pregnant rats receiving salted-water were compared to not-supplemented pregnant rats. Markers of oxidative stress, ensuing cell death, and changes in the production of vasoactive substances (prostanoids: thromboxane, TxB(2); and prostacyclin, PGF(1alpha)) and the pro-inflammatory cytokine tumour necrosis factor-alpha (TNF-alpha) were measured in the placenta. In tissue from pregnant rats on 1.8% NaCl supplement, 8-iso-PGF(2alpha) levels, TxB(2)/6-keto-PGF(1alpha) ratios, total TNF-alpha RNA expression, as well as the apoptotic index (Bax/Bcl-2 ratio) and endothelial nitric oxide synthase protein expression increase while total glutathione content decreases. These findings demonstrate that maternal insult during gestation induced an imbalance in the oxidative environment in the placenta favouring oxidation. This was accompanied by an increased synthesis of vasoconstrictive substances and TNF-alpha by the placenta as well as the increased rate of placental cell apoptosis. PMID- 16469377 TI - Translocation (13;17)(q14;q25) as a novel chromosomal abnormality in acute myeloid leukemia-M4. AB - We report a case of AML-M4 in which G-band karyotyping revealed a previously unreported t(13;17)(q14;q25) in metaphase preparations. The breakpoints at 13q14 and 17q25 are associated with poor prognosis. The MSF and FKHR genes are located on 17q25 and 13q14, respectively. This report of AML-M4 harboring t(13;17)(q14;q25) as a unique cytogenetic abnormality provides more data on the leukomogenesis with rearrangements related with 13q14 and 17q25. PMID- 16469378 TI - Heavy metal contamination and antioxidant response of a freshwater bryozoan (Lophopus crystallinus Pall., Phylactolaemata). AB - Monthly variations of total glutathione, glutathione-S-transferases, glutathione peroxidase, glutathione reductase, glyoxalase I, glyoxalase II, and catalase were studied in Lophopus crystallinus, a freshwater bryozoan, collected from Lake Piediluco. Specimens were sampled during the vegetative period from November to April for two different cycles: cycle I from 1996 to 1997 and cycle II from 1999 to 2000. Simultaneously, analyses of heavy metal content such as lead, cadmium, chromium, nickel, copper, iron, and zinc were determined in the samples of L. crystallinus. Specimens of cycle II displayed highest chromium and iron accumulation and elevated levels of biochemical parameters. Total glutathione content, catalase, and glutathione peroxidase activities, were related with the highest metal concentrations in L. crystallinus from Lake Piediluco. PMID- 16469379 TI - Assessing the aquatic hazard of commercial hydrocarbon resins. AB - Hydrocarbon resins are used to modify polymer products to achieve desired functional properties for a diverse range of products. These complex hydrocarbon based mixtures are typically poorly soluble in water. However, resins may leach lower-molecular-weight monomers or impurities upon contact with water, thus posing a potential hazard to the aquatic environment. The bioavailability and toxicity of leachable constituents of four solid and three liquid resins were evaluated by analyzing water-accommodated fractions prepared with each resin, using biomimetic solid phase microextraction (SPME) techniques. Liquid resins exhibited concentrations of bioavailable constituents that were sufficiently elevated to cause acute toxicity to the aquatic organism Daphnia magna. All solid resins exhibited lower bioavailable concentrations of leachable constituents that were unlikely to pose an aquatic toxicity concern. Since observed toxicity of both resin types was generally consistent with bioavailable concentrations determined using SPME fiber measurements, it is concluded that this approach provides a convenient in vitro screening tool that can help reduce the use of animal testing in environmental hazard assessment of complex hydrocarbon-based substances. PMID- 16469380 TI - Crucian carp (Carassius carassius) VTG monoclonal antibody: development and application. AB - The vitellogenin (VTG) in fish has been used as an important biomarker for monitoring endocrine disrupting compounds (EDCs). This paper reports the development of a new monoclonal antibody (McAb) against the VTG of crucian carp (Carassius carassius). The McAb has a molecular weight of 149.4 kDa (heavy chain: 53.1 kDa; light chain: 21.6 kDa), and double diffusion indicated that it belongs to the IgG1 subclass. The titer is 10(5)-10(6) and the affinity constant (K(aff)) is 7.0 x 10(8)L/mol, showing the high specificity and sensitivity of the antibody. The established sandwich ELISA was sensitive with a detection limit of VTG 0.98 ng/mL. The cross-reactivity of antibody was detected in cyprinids such as rare minnow, zebrafish, and carp. This ELISA was used to detect the variation of VTG in crucian carp exposed to secondary effluent and reclaimed water from the Gaobeidian sewage treatment plant (SRP). The VTG induction in secondary effluent was higher than that in reclaimed effluent and the VTG levels in juvenile crucian carp increased with increasing exposure time. The VTG concentration in male fish from downstream of the Gaobeidian STP was 88.62+/-827.73 microg/mL, while that from a control site was undetectable. PMID- 16469381 TI - Representing revolution: icons of industrialization. AB - Appreciating pictures entails a consideration not only of the people, objects and landscape that their artists have chosen to portray, but also an imagining of what has been excluded. The term 'Industrial Revolution' has been given multiple meanings, and this article (part of the Science in the Industrial Revolution series) explores some of these by exposing the messages concealed inside some of the most enduring images of the Revolution. PMID- 16469383 TI - The origins of research into the origins of life. AB - Most scientists at the end of the 19th and the beginning of the 20th century chose to ignore the question of the origin of life on Earth, regarding it as too mysterious and complex to handle. Yet, in the early 1950s an experimental field devoted to the study of the problem made its first steps. The pioneering theories of several scientists in the first decades of the 20th century played a major role in this transformation, notably those of the Russian biochemist Alexander I. Oparin and the British geneticist and biochemist J.B.S. Haldane. The ideas of the lesser-known American psycho-physiologist Leonard Troland also made a significant contribution to subsequent developments in origin-of-life research. Therefore, it is well worth taking a look at the professional, philosophical and ideological commitments that shaped the approaches of the three scientists to origin-of-life research. PMID- 16469384 TI - Cross-presentation of a human malaria CTL epitope is conformation dependent. AB - Little is known about the role of conformation on the antigen processing by antigen presenting cells. Using a well-defined antigen containing two disulfide bridges, the synthetic C-terminal fragment 282-383 derived from Plasmodium falciparum circumsporozoite protein (PfCS 282-383), we show that the reduced form is presented in vitro more efficiently than its oxidized counterpart, inducing stronger CTL recognition. In addition, only the reduced form can be presented by the TAP independent T2 cell line. Thus, the reduced form is processed by TAP dependent and independent pathways. PMID- 16469385 TI - A(1) adenosine receptor signal and AMPK involving caspase-9/-3 activation are responsible for adenosine-induced RCR-1 astrocytoma cell death. AB - Extracellular adenosine reduced viability of RCR-1 rat astrocytoma cells in a dose (0.3-10mM)- and treatment time (24-72h)-dependent manner. In the apoptosis assay using propidium iodide (PI) and annexin V, treatment with adenosine (1mM) for 72h increased the population of PI-negative/annexin V-positive cells, that is related to early apoptosis, and that of PI-positive/annexin V-positive cells, that is related to late apoptosis/secondary necrosis. In addition, nuclei of cells treated with adenosine (1mM) for 72h were reactive to an antibody against single-stranded DNA. Adenosine activated caspase-3, -8 and -9, but mitochondrial membrane potentials were not affected. Adenosine-induced RCR-1 cell death was significantly inhibited by 8-CPT, an antagonist of A(1) adenosine receptors, and forskolin, an adenylate cyclase activator. SQ22536, an adenylate cyclase inhibitor, alternatively, exhibited an effect similar to adenosine. CHA, an agonist of A(1) adenosine receptors, activated caspase-3 and -9, but not caspase 8. Adenosine-induced cytotoxicity of RCR-1 cells was also significantly inhibited by dipyridamole, an inhibitor of adenosine transporter, and AMDA, an inhibitor of adenosine kinase. AICAR, an activator of AMP-activated protein kinase (AMPK), reduced RCR-1 cell viability, but synergistic effect was not obtained with co treatment with adenosine and AICAR. AICAR activated caspase-3 and -9, but not caspase-8. An additive inhibition was found in the co-presence of 8-CPT and dipyridamole. Extracellular adenosine, thus, appears to activate caspase-9 followed by the effector caspase, caspase-3, at least via two independent pathways linked to A(1) adenosine receptor-mediated adenylate cyclase inhibition and adenosine uptake into cells/conversion to AMP/activation of AMPK, possibly regardless of mitochondrial damage, thereby leading to RCR-1 cell death, dominantly by apoptosis. Moreover, caspase-8 activation could again contribute to adenosine-induced cytotoxicity, although the underlying mechanism is currently unknown. Collectively, the results of the present study may represent a new pathway for caspase activation relevant to diverse adenosine signals in cell death. PMID- 16469387 TI - The centrifugal visual system of vertebrates: a comparative analysis of its functional anatomical organization. AB - The present review is a detailed survey of our present knowledge of the centrifugal visual system (CVS) of vertebrates. Over the last 20 years, the use of experimental hodological and immunocytochemical techniques has led to a considerable augmentation of this knowledge. Contrary to long-held belief, the CVS is not a unique property of birds but a constant component of the central nervous system which appears to exist in all vertebrate groups. However, it does not form a single homogeneous entity but shows a high degree of variation from one group to the next. Thus, depending on the group in question, the somata of retinopetal neurons can be located in the septo-preoptic terminal nerve complex, the ventral or dorsal thalamus, the pretectum, the optic tectum, the mesencephalic tegmentum, the dorsal isthmus, the raphe, or other rhombencephalic areas. The centrifugal visual fibers are unmyelinated or myelinated, and their number varies by a factor of 1000 (10 or fewer in man, 10,000 or more in the chicken). They generally form divergent terminals in the retina and rarely convergent ones. Their retinal targets also vary, being primarily amacrine cells with various morphological and neurochemical properties, occasionally interplexiform cells and displaced retinal ganglion cells, and more rarely orthotopic ganglion cells and bipolar cells. The neurochemical signature of the centrifugal visual neurons also varies both between and within groups: thus, several neuroactive substances used by these neurons have been identified; GABA, glutamate, aspartate, acetylcholine, serotonin, dopamine, histamine, nitric oxide, GnRH, FMRF-amide-like peptides, Substance P, NPY and met-enkephalin. In some cases, the retinopetal neurons form part of a feedback loop, relaying information from a primary visual center back to the retina, while in other, cases they do not. The evolutionary significance of this variation remains to be elucidated, and, while many attempts have been made to explain the functional role of the CVS, opinions vary as to the manner in which retinal activity is modified by this system. PMID- 16469388 TI - Semi-automated quantification of axonal densities in labeled CNS tissue. AB - Current techniques used to quantify axons often rely upon manual quantification or potentially expensive commercially available programs for automated quantification. We describe a computerized method for the detection and quantification of axons in the rat CNS using readily available free software. Feature J, a java-based plug-in to the imaging software NIH Image J, faithfully detects linear structures such as axons in confocal or bright-field images using a Hessian-based algorithm. We validated the method by comparing values obtained by manual and automated analyses of axons induced to grow in response to neurotrophin over-expression in the rat spinal cord. We also demonstrated that the program can be used to quantify neurotrophin-induced growth of lesioned serotonergic axons in the rat cortex, where manual measurement would be impractical due to dense axonal growth. The use of this software suite provided faster and less biased quantification of labeled axons in comparison to manual measurements at no cost. PMID- 16469389 TI - Cognitive impairment in bipolar disorder in old age: literature review and findings in manic patients. AB - BACKGROUND: Descriptions of aged patients with bipolar (BP) disorder have commented on cognitive impairments. However, the literature regarding cognitive test performance in this population has apparently been scant. METHOD: 1. We reviewed studies reporting cognitive performance in aged BP patients. 2. We compared the performance of elderly BP manic patients and aged community comparison subjects on the Mini-Mental State Examination (MMSE) and the Mattis Dementia Rating Scale (DRS). RESULTS: 1. Seven published studies of cognitive measures in aged BP patients were identified. They utilized different assessment methods and addressed different illness states, but they indicate impairments in these patients. 2. In our sample, the manic patients (n=70) had lower MMSE scores and DRS scores than did the comparison subjects (n=37). In these patients, cognitive scores were not significantly associated with Mania Rating Scale scores. LIMITATIONS: The patients in our study were assessed cross-sectionally, and they were treated naturalistically. CONCLUSIONS: Manic or depressed BP elders have impaired cognitive function; in some patients these impairments may persist. Research characterizing these impairments and their clinical implications is warranted. PMID- 16469386 TI - Oxidative dealkylation DNA repair mediated by the mononuclear non-heme iron AlkB proteins. AB - DNA can be damaged by various intracellular and environmental alkylating agents to produce alkylation base lesions. These base damages, if not repaired promptly, may cause genetic changes that lead to diseases such as cancer. Recently, it was discovered that some of the alkylation DNA base damage can be directly removed by a family of proteins called the AlkB proteins that utilize a mononuclear non-heme iron(II) and alpha-ketoglutarate as cofactor and cosubstrate. These proteins activate dioxygen and perform an unprecedented oxidative dealkylation of the alkyl adducts on DNA heteroatoms. This review summarizes the discovery of this activity and the recent research advances in studying this unique DNA repair pathway. The focus is placed on the chemical mechanism and function of these proteins. PMID- 16469390 TI - Glial cell number and neuron/glial cell ratios in postmortem brains of bipolar individuals. AB - BACKGROUND: There are several lines of evidence implicating a glial abnormality in the pathophysiology of bipolar disorder. Previous studies have reported a wide range of abnormalities but with little consistency between their findings. METHODS: Sixty frozen postmortem temporal cortical brain samples from normal, schizophrenic, bipolar, and depressed subjects were obtained from the Stanley Neuropathology Consortium. Nissl stain was utilized to visualize cellular structures. The sections were analyzed for neuron and glial content using the computer programs SPOT Advanced and Metamorph. Three representative fields from each subject were counted and the average numbers obtained. RESULTS: There was no critical difference in glia number across the four diagnostic groups. Compared with normal controls, area occupied by glia was reduced in bipolar subjects (P = 0.018), and the ratio of glial area to neuronal area was reduced (P = 0.028). DISCUSSION: The percent glial space was substantially lower in bipolar disorder subjects compared to normal controls, suggesting that the glia may be smaller in bipolar subjects. The size reduction may be reflective of glial dysfunction. LIMITATIONS: Postmortem interval of 29.4 +/- S.D. 13.4 h may have affected cellular structure and reduced the quality of the staining. PMID- 16469391 TI - A shared neuroimmunological mechanism for drugs of addiction? PMID- 16469393 TI - Complications in paediatric airway management--pneumothorax. PMID- 16469392 TI - Monoclonal MOG-reactive autoantibody from progressive EAE has the characteristics of a natural antibody. AB - A.SW mice sensitized with myelin oligodendrocyte glycoprotein (MOG)92-106 is an animal model for progressive multiple sclerosis (MS). We isolated MOG-reactive monoclonal antibodies that were immunoglobulin (Ig)M and polyreactive, similar to natural autoantibodies. Upon analysis of the variable (V) light chains and the diversity (D) and joining (J) regions of V heavy chains, we found they were identical to germ line Vkappa19/28, Jkappa5, DFL16.1e and JH4, respectively. The sequence of the VH region had 99.7% and 100% identity at the nucleotide and amino acid levels, respectively, compared with the germ line encoded antibody, P3, of the Q52 family. Although A strain mice have been reported to have an insertion in BAFF-R, the receptor for BAFF (B cell activation factor from the tumor necrosis factor family), which could explain our results, A.SW mice have no mutations in BAFF-R. PMID- 16469394 TI - Late postnatal onset of hearing loss due to GJB2 mutations. AB - GJB2 mutations account for approximately 50% of recessive non-syndromic deafness, with 35delG being the most prevalent. Homozygous 35delG mutations cause pre lingual, non-progressive hearing loss that is detected on newborn hearing screening programmes. We present a sibling pair with homozygous 35delG mutations, who passed hearing tests in early infancy and developed progressive sensorineural hearing loss, one requiring a cochlear implant. These cases illustrate that deafness due to such mutations may have a late onset and consequently be missed on neonatal screening programmes and they may present an argument to consider neonatal screening for GJB2 mutations in order to aid early intervention. PMID- 16469395 TI - A comparison of Lewis and Fischer rat strains on autoshaping (sign-tracking), discrimination reversal learning and negative auto-maintenance. AB - Lewis (LEW) and Fischer (F344) rat strains differ on a number of physiological characteristics, such as hypothalamic-pituitary-adrenal (HPA) axis activity, as well as on behavioral tasks, including those that measure impulsivity and drug reward. Since autoshaping, the phenomenon where animals approach and contact reward-paired conditioned stimuli, has been linked to HPA axis functioning, impulsivity and drug taking, the present study compared LEW and F344 rats on the rate of acquisition and performance of the autoshaping response. Rats were trained on an autoshaping procedure where insertions of one retractable lever (CS(+)) were paired response-independently with food, while insertions of another lever (CS(-)) were not paired with food. LEW rats acquired the autoshaping response more rapidly and also performed the autoshaping response at a higher rate than F344 rats. No differences between the strains were observed when rats were trained on a discrimination reversal where the CS(+) and CS(-) levers were reversed or during a negative auto-maintenance phase where CS(+) lever contacts cancelled food delivery. Potential physiological mechanisms that might mediate the present results, including strain differences in HPA axis and monoamine neurotransmitter activity, are discussed. The finding that LEW (as compared to F344 rats) more readily acquire autoshaping and perform more responses is consistent with research indicating that LEW rats behave more impulsively and more readily self-administer drugs of abuse. PMID- 16469396 TI - A novel 78-kDa fatty acyl-CoA synthetase (ACS1) of Babesia bovis stimulates memory CD4+ T lymphocyte responses in B. bovis-immune cattle. AB - Antigen-specific CD4+ T lymphocyte responses contribute to protective immunity against Babesia bovis, however the antigens that induce these responses remain largely unknown. A proteomic approach was used to identify novel B. bovis antigens recognized by memory CD4+ T cells from immune cattle. Fractions obtained from merozoites separated by continuous-flow electrophoresis (CFE) that contained proteins ranging from 20 to 83 kDa were previously shown to stimulate memory CD4+ lymphocyte responses in B. bovis-immune cattle. Expression library screening with rabbit antiserum raised against an immunostimulatory CFE fraction identified a clone encoding a predicted 78 kDa protein. BLAST analysis revealed sequence identity of this B. bovis protein with Plasmodium falciparum fatty acyl coenzyme A synthetase (ACS) family members (PfACS1-PfACS11), and the protein was designated B. bovis acyl-CoA synthetase 1 (ACS1). Southern blot analysis indicated that B. bovis ACS1 is encoded by a single gene, although BLAST analysis of the preliminary B. bovis genome sequence identified two additional family members, ACS2 and ACS3. Peripheral blood lymphocytes and CD4+ T cell lines from B. bovis-immune cattle proliferated significantly against recombinant ACS1 protein, consistent with its predicted involvement in protective immunity. However, immune sera from cattle recovered from B. bovis infection did not react with ACS1, indicating that epitopes may be conformationally dependent. PMID- 16469397 TI - FTF and LRH-1, two related but different transcription factors in human Caco-2 cells: their different roles in the regulation of bile acid transport. AB - The apical sodium dependent bile acid transporter (ASBT) mediates ileal bile acid reabsorption. The transcription factors, liver receptor homologue-1 (LRH-1:mouse) and fetoprotein transcription factor (FTF:human), are presumably orthologues. Bile-acid induced negative feedback regulation of mouse (m) and human (h) ASBT occurs via LRH-1 and RAR/RXR, respectively. hASBT has a potential FTF cis element, although its functional role is unknown. hASBT and mASBT promoter constructs and an FTF cis-element mutated hASBT (hASBT/FTFmu) were assessed in human Caco-2 cells treated with chenodeoxycholic acid (CDCA) and/or co transfected with hFTF, mLRH-1, or specific small interfering FTF or LRH-1 RNA (siFTF or siLRH). Basal promoter activity was reduced in hASBT/FTFmu, although bile acid response persisted. hFTF activated hASBT but not mASBT, while mLRH-1 activated mASBT but not hASBT. siFTF reduced hASBT but not mASBT activity; siLRH reduced mASBT but not hASBT activity. siLRH but not siFTF abrogated bile acid responsiveness. Electrophoretic mobility shift assays demonstrated distinct and specific binding of the mLRH-1 or hFTF cis-elements. In conclusion, FTF and LRH-1 are two related but different transcription factors in human Caco-2 cells, suggesting that they may be homologues and not orthologues. FTF is not involved directly in bile acid mediated negative feedback regulation of the ASBT. PMID- 16469398 TI - Zebrafish z-otu, a novel Otu and Tudor domain-containing gene, is expressed in early stages of oogenesis and embryogenesis. AB - Several studies have suggested that Otu domain had de-ubiquitinating activity and Tudor domain was important for the formation of germ cells. Here, we reported a novel zebrafish ovary-specific gene containing Otu and Tudor domain, z-otu, which was expressed at stages I-III oocytes and embryonic stages from zygotes to early blastula during embryonic cells maintained their totipotency. Therefore, z-otu might link the ubiquitin signaling pathway to early oogenesis and maintaining the totipotency of embryonic cell. PMID- 16469399 TI - The measurement of change in functional ability: dealing with attrition and the floor/ceiling effect. AB - The purpose was to describe four-year change in functional ability among older persons and the relationship to sex, age, and other background factors. The baseline study, performed in 1986, is based on a random sample of older persons (n=1261). Follow-up data were collected four-years later (n=912). The analyses of change in functional ability were based on the assumption that the categories reflected an underlying latent continuous dimension. The change in functional ability, DeltaFA, was calculated by a logistic model for paired observations and applied in parallel analyses with and without inclusion of the dead to deal with the attrition problem. Fifty percent had no change in functional ability, 37% had declined and 13% improved. Models including the dead showed more functional decline with increasing age but this was not the case when the dead were excluded. Functional change was not related to sex, functional ability at baseline, relative wealth, social network, self-rated health, and life satisfaction. Inclusion of the dead in statistical models for the study of change in functional ability reduced the attrition problem. A logistic model for paired observations of functional ability at two points in time reduced the problem related to the floor/ceiling problem. PMID- 16469400 TI - Use of competitive DNA hybridization to identify differences in the genomes of bacteria. AB - Although recent technological advances in DNA sequencing and computational biology now allow scientists to compare entire microbial genomes, comparisons of closely related bacterial species and individual isolates by whole-genome sequencing approaches remains prohibitively expensive for most laboratories. Here we report the development and testing of a biochemical approach for targeted sequencing of only those chromosomal regions that differ between two DNA preparations. The method, designated GFE (genome fragment enrichment) uses competitive solution hybridization and positive selection to obtain genomic DNA fragments that are present in one pool of fragments but not another. Repeated comparisons of the genomes of Enterococcus faecalis and E. faecium led to the identification of 225 putative genome-specific DNA fragments. Species and strain variations within these fragments were confirmed by both experimental and bioinformatic analyses. The E. faecalis genome-specific sequences identified included both a preponderance of those predicted to encode surface-exposed proteins, as well as several previously described unique marker regions embedded within highly conserved rrn operons. The GFE strategy we describe efficiently identified genomic differences between two enterococcal genomes, and will be widely applicable for studying genetic variation among closely related bacterial species. PMID- 16469401 TI - Gene expression array analyses predict increased proto-oncogene expression in MMTV induced mammary tumors. AB - Exogenous infection by milk-borne mouse mammary tumor viruses (MMTV) typically induce mouse mammary tumors in genetically susceptible mice at a rate of 90-95% by 1 year of age. In contrast to other transforming retroviruses, MMTV acts as an insertional mutagen and under the influence of steroid hormones induces oncogenic transformation after insertion into the host genome. As these events correspond with increases in adjacent proto-oncogene transcription, we used expression array profiling to determine which commonly associated MMTV insertion site proto oncogenes were transcriptionally active in MMTV induced mouse mammary tumors. To verify our gene expression array results we developed real-time quantitative RT PCR assays for the common MMTV insertion site genes found in RIII/Sa mice (int 1/wnt-1, int-2/fgf-3, int-3/Notch 4, and fgf8/AIGF) as well as two genes that were consistently up regulated (CCND1, and MAT-8) and two genes that were consistently down regulated (FN1 and MAT-8) in the MMTV induced tumors as compared to normal mammary gland. Finally, each tumor was also examined histopathologically. Our expression array findings support a model whereby just one or a few common MMTV insertions into the host genome sets up a dominant cascade of events that leave a characteristic molecular signature. PMID- 16469402 TI - Mimivirus and the emerging concept of "giant" virus. AB - The recently discovered Acanthamoeba polyphaga Mimivirus is the largest known DNA virus. Its particle size (750 nm), genome length (1.2 million bp) and large gene repertoire (911 protein coding genes) blur the established boundaries between viruses and parasitic cellular organisms. In addition, the analysis of its genome sequence identified many types of genes never before encountered in a virus, including aminoacyl-tRNA synthetases and other central components of the translation machinery previously thought to be the signature of cellular organisms. In this article, we examine how the finding of such a giant virus might durably influence the way we look at microbial biodiversity, and lead us to revise the classification of microbial domains and life forms. We propose to introduce the word "girus" to recognize the intermediate status of these giant DNA viruses, the genome complexity of which makes them closer to small parasitic prokaryotes than to regular viruses. PMID- 16469403 TI - [Management of adhesive capsulitis of the shoulder]. PMID- 16469404 TI - Antioxidants versus corticosteroids in the treatment of severe alcoholic hepatitis--a randomised clinical trial. AB - BACKGROUND/AIMS: Severe alcoholic hepatitis is associated with high morbidity and short-term mortality. Corticosteroids are the only widely used therapy but established contraindications to treatment or the risk of serious side-effects limit their use. The perceived need for alternative treatments together with the theoretical benefits of anti-oxidant therapy triggered the design of a randomised clinical trial comparing these treatment modalities. METHODS: One hundred and one patients were randomized into a clinical trial of corticosteroids or a novel antioxidant cocktail with a primary endpoint of 30-day mortality. RESULTS: At 30 days there were 16 deaths (30%) in the corticosteroid treated group compared with 22 deaths (46%) in the antioxidant treated group (P=0.05). The odds of dying by 30 days were 2.4 greater for patients on antioxidants (95% confidence interval 1.0-5.6). A diagnosis of sepsis was made more frequently in the AO group (P=0.05), although microbiologically proven episodes of infection occurred more often in the CS group (P<0.01). The survival advantage for corticosteroid treated patients was lost at 1 year of follow-up (P=0.43). CONCLUSIONS: This study has shown that corticosteroids in the form of prednisolone 30 mg daily are superior to a broad antioxidant cocktail in the treatment of severe alcoholic hepatitis. PMID- 16469405 TI - Growth and differentiation of colony-forming human hepatocytes in vitro. AB - BACKGROUND/AIMS: Parenchymal hepatocytes (PHs) of rat contain colony-forming parenchymal hepatocytes (CF-PHs) as a small fraction. We aimed to demonstrate the presence of CF-PHs in humans and characterize them with respect to growth and differentiation potential. METHODS: Human PHs were co-cultured with Swiss 3T3 cells in the medium containing human serum, EGF, nicontinamide, and ascorbic acid 2-phosphate. To examine differentiation potential hepatocytes were cultured on gels of Matrigel Matrix. RESULTS: Few PHs formed colonies, the colony-forming efficiency being as low as 0.01-0.09%. The CF-PHs could be subcultured up to 7 passages. They showed a liver epithelial cell-like morphology, and immunocytochemically positive for albumin (ALB), cytokeratin (CK) 7, 8, 18, and 19 in a pre- and early phase-confluence, whereas they showed a typical differentiated hepatocyte-like morphology, and positive for alpha(1)-antitrypsin, but negative for CK7 and 19 in condensed regions at confluence. The CF-PHs at late confluence expressed mRNAs of ALB, HNF4, and isoforms of cytochrome P450 at low levels. However, when cultured on Matrigel, these cells expressed them at high levels comparable to those of original PHs. CONCLUSIONS: We concluded that the human liver contains highly replicative hepatic progenitor-like cells as a minute population that retain a normal differentiation potential. PMID- 16469406 TI - Hammerhead ribozymes with cleavage site specificity for NUH and NCH display significant anti-hepatitis C viral effect in vitro and in recombinant HepG2 and CCL13 cells. AB - BACKGROUND/AIMS: Four different ribozymes (Rz) targeting the hepatitis C virus (HCV) 5'-non-coding region (NCR) at nucleotide (nt) positions GUA 165 (Rz1), GUC 270 (Rz2), GUA 330 (Rz3) and GCA 348 (Rz1293) were compared for in vitro cleavage using a 455 nt HCV RNA substrate. The GUA 330 (Rz3) and GCA 348 (Rz1293) ribozymes, both targeting the HCV loop IV region, were found to be the most efficient, and were further analyzed in an in vitro translation system. METHODS: For this purpose RNA transcribed from a construct encoding a HCV-5'-NCR luciferase fusion protein was used. Cleavage-inactive (Rz1426), mismatch (Rz1293m) or unrelated ribozymes (Rz1437) were synthesized as controls for Rz 1293. HCV specificity was analysed by competition experiments using sense and mismatch oligodeoxynucleotides HCVrzCI and HCVrzMM, respectively. RESULTS: A chemically modified nuclease-resistant variant of the GCA 348 cleaving ribozyme was selected for cell culture experiments using recombinant HepG2 or CCL13 cell lines stably transfected with a HCV-5'-NCR-luciferase target construct. CONCLUSIONS: This ribozyme (Rz1293) showed an inhibitory activity of translation of more than 70% thus verifying that the GCA 348 cleavage site in the HCV loop IV is an accessible target site in vivo and may be suitable for the development of novel optimized hammerhead structures. PMID- 16469408 TI - Therapeutic effect of transplanting HGF-treated bone marrow mesenchymal cells into CCl4-injured rats. AB - BACKGROUND/AIMS: The autologous transplantation of bone marrow cells is a promising treatment for liver disease. Pluripotent bone marrow stem cells can differentiate into hepatocytes, but few reports address the therapeutic effect of transplanting these stem cells into damaged liver in vivo. Here, we transplanted bone marrow-derived mesenchymal cells (BMMCs) to test their effect in liver injured rats. METHODS: Rat bone marrow cells were cultivated for 2 weeks in the presence or absence of hepatocyte growth factor (HGF), labeled with a fluorescent marker, and transplanted by injection into CCl(4)-injured rats. Blood samples collected 4 weeks later were analyzed for albumin production and transaminase levels. The amount of fibrosis was determined by histology. RESULTS: RT-PCR analysis detected alpha-fetoprotein and albumin mRNAs in BMMCs cultured with HGF for 2 weeks. Albumin protein was also produced in the BMMC cultures by a subpopulation of cells. Transplantation of the BMMCs into liver-injured rats restored their serum albumin level and significantly suppressed transaminase activity and liver fibrosis. These effects were not seen when the BMMCs were cultured without HGF. CONCLUSIONS: The transplantation of BMMCs cultured with HGF effectively treats liver injury in rats. This is a promising technique for autologous transplantation in humans with liver injury. PMID- 16469409 TI - A secure web-based approach for accessing transitional health information for people with traumatic brain injury. AB - A web-based transitional health record was created to provide regional healthcare professionals with ubiquitous access to information on people with brain injuries as they move through the healthcare system. Participants included public, private, and community healthcare organizations/providers in Eastern Ontario (Canada). One hundred and nineteen service providers and 39 brain injury survivors registered over 6 months. Fifty-eight percent received English and 42% received bilingual services (English-French). Public health providers contacted the regional service coordinator more than private providers (52% urban centres, 26% rural service providers, and 22% both areas). Thirty-five percent of contacts were for technical difficulties, 32% registration inquiries, 21% forms and processes, 6% resources, and 6% education. Seventeen technical enquiries required action by technical support personnel: 41% digital certificates, 29% web forms, and 12% log-in. This web-based approach to clinical information sharing provided access to relevant data as clients moved through or re-entered the health system. Improvements include automated digital certificate management, institutional health records system integration, and more referral tracking tools. More sensitive test data could be accessed on-line with increasing consumer/clinician confidence. In addition to a strong technical infrastructure, human resource issues are a major information security component and require continuing attention to ensure a viable on-line information environment. PMID- 16469407 TI - Over-expression of interleukin-6 enhances cell survival and transformed cell growth in human malignant cholangiocytes. AB - BACKGROUND/AIMS: Over-expression of IL-6 has been implicated in cholangiocarcinoma growth but the cellular mechanisms involved are unknown. Our aims were to assess the mechanisms by which over-expression of IL-6 promotes transformed cell growth in malignant cholangiocytes. METHODS: Stably transfected cell lines over-expressing IL-6 were derived from malignant human cholangiocytes. Transformed cell growth was assessed by anchorage independent growth in vitro and by xenograft growth in nude mice. Expression of the anti-apoptotic protein Mcl-1 was quantitated by immunoblot analysis and by real-time PCR. Gene silencing was performed using siRNA. Dominant negative upstream kinase activators and isoform specific constructs were used to evaluate the involvement of p38 MAP kinase signaling pathways. RESULTS: Over-expression of IL-6 increased xenograft growth, anchorage independent growth and cell survival but did not significantly alter cell proliferation. The basal expression of Mcl-1 was increased in IL-6 over expressing cells. Selective knockdown of Mcl-1 by siRNA increased gemcitabine induced cytotoxicity. Moreover, IL-6 increased Mcl-1 mRNA and protein expression via a p38 MAPK dependent mechanism. CONCLUSIONS: These data demonstrate a major role of survival signaling pathways in mediating the effects of IL-6 over expression in cholangiocarcinoma growth. Mcl-1 is identified as a mediator of IL 6-induced tumor cell survival and shown to be transcriptionally regulated by IL-6 via a p38 MAPK dependent pathway. We conclude that modulation of IL-6 mediated survival signaling pathways involving the p38 MAPK or downstream targets such as Mcl-1 may prove useful therapeutic strategies for human cholangiocarcinoma. PMID- 16469410 TI - Long-duration, weekly treatment with gemcitabine plus vinorelbine for non-small cell lung cancer: a multicenter phase II study. AB - In this phase II study, gemcitabine and vinorelbine were combined at suboptimal doses for weekly administration in advanced non-small cell lung cancer (NSCLC). The primary objectives were to determine objective response rate (ORR) and time to progression (TTP). Secondary endpoints were safety and overall survival. Chemonaive patients with histologically or cytologically confirmed stage IIIB or IV NSCLC received vinorelbine (25 mg/m2) immediately followed by gemcitabine (800 mg/m2) once each week (on day 1) for 6 months without rest. From May 1998 to May 1999, 40 patients were enrolled (85% males; 70% stage IV) with a median age of 65.5. A total of 478 doses were administered, with a median of 9 per patient (range 2-72). The ORR was 27.5% (95% CI, 15.1-44.1%). The median TTP was 3.5 months (95% CI, 2.9-4.4 months). At a median follow-up of 6.5 months, the median survival was 11.6 months, and survival rates at 1 and 2 year(s) were 47.5% and 15.8%, respectively. The most common grade 3/4 hematologic toxicity was neutropenia, in 70% of patients, with febrile neutropenia in 28%. The most common grade 3/4 non-hematologic toxicity was transaminase elevation, in 22.5% of patients, which was transient and reversible. The other most prominent toxicities were, unexpectedly, pulmonary and cardiac toxicities. Based on these results, weekly, long-term administration of gemcitabine-vinorelbine appears to be an active regimen in NSCLC that warrants further investigation. PMID- 16469411 TI - Does high intake of fruit and vegetables improve lung cancer survival? AB - The objective of the study was to examine the prognostic effect of dietary intake of fruit and vegetables on lung cancer patients. We used data on 57,053 participants in the Danish prospective cohort study, 'Diet, Cancer and Health'. Patients in whom lung cancer was diagnosed constituted the final study cohort and were followed from the date of diagnosis until the date of death or 11 March 2004. A total of 353 participants had lung cancer. Increasing levels of intake of fruit and vegetables show a tendency toward decreased hazard of dying: the Cox proportional hazard model estimated a hazard ratio (HR) of 0.84 (95% confidence interval [CI], 0.59-1.21) for high intake of vegetables by current smokers and an HR of 0.81 (95% CI, 0.58-1.15) for high intake of fruits with low intake as the reference. In contrast, high intake of potatoes increased the hazard of dying (HR, 1.51; 95% CI, 1.12-2.23). Our study suggests that high intake of fruit and vegetables might have a favourable effect on the prognosis of lung cancer patients, but a high intake of potatoes appears to increase the hazard of dying. PMID- 16469412 TI - Sorption of naphthoic acids and quinoline compounds to estuarine sediment. AB - The sorption of 16 ionizable organic compounds (IOCs) to an estuarine sediment was measured in synthetic estuarine water as a function of IOC concentration (1 100 microM) at fixed ionic strength (0.4 M), pH (7.6), and sediment concentration (0.018 g sediment kg(-1) suspension). Of the 16 IOCs, 11 were naphthoic acids and five were quinoline compounds. The linear sorption distribution coefficient (Kd) was used to correlate sorption to IOC physicochemical and molecular characteristics. With respect to naphthoic acid, sorption increased with the addition of ortho-substituent groups and with increasing chain length of the 1 acid group, and the greatest increase occurred with ortho-hydroxyl, carbonyl, and carboxyl groups. With respect to quinoline, sorption decreased with substituent group addition (except for nitro group) and with additional heterocyclic N atoms. For the naphthoic acids, log Kd exhibited a positive correlation with water solubility (log Sw) indicative of sorption primarily to mineral surfaces under the solution chemistry. For the quinoline compounds, log Kd exhibited a negative correlation with log Sw and a positive correlation with n-octanol/water partition coefficient (log K(OW)) indicative of sorption primarily to organic matter. For both compounds, poor or no correlations were established between log Kd and acid dissociation constant (pKa1), and between log Kd and a variety of molecular connectivity indexes. The results from this study demonstrate that the sorption of IOCs differ depending on their backbone structure and may differ between parent compound and ionizable degradation product. PMID- 16469413 TI - Heart disease among post-menopausal women: acceptability of functional foods as a preventive measure. AB - This study reports the results of eight focus group interviews with post menopausal women (50-59 years, N=73) in Denmark and United Kingdom regarding their views on heart disease among women and the potential acceptability of functional foods as a means of prevention. Although, heart disease was recognised as a serious health problem and its occurrence among women recognised, it was predominantly conceived as a health problem affecting men and as being largely due to the relatively unhealthy lifestyles of men. Only a very small minority of participants were aware of the contribution of menopause to heart disease among women. Functional foods were generally not conceived as a means of prevention that is commensurate with the serious character of heart disease. It emerged from ranking tasks that functional foods do not constitute a product category as such. They occupy an anomalous position between 'food' on the one hand and 'medicine' on the other. Participants tend to dislike the idea of a 'personal' food not intended to comprise part of a shared meal in the household, and also tend to distrust health claims promoted by the food industry. This study indicates a need for further research in these areas. PMID- 16469414 TI - Awareness, acceptance of and willingness to buy genetically modified foods in Urban China. AB - There is concern about the extent to which consumers will accept genetically modified (GM) foods if they are commercialized in China. The evidence from the existing literature is mixed and sometimes confusing. The objective of this study is to conduct a large in-depth face-to-face in-house survey that examines the consumers' awareness, acceptance of and willingness to buy GM foods in China. To achieve this objective, a well-designed consumer survey was conducted in 11 cities of five provinces in Eastern China in 2002 and 2003. The results indicate that despite much less information on GM foods available publicly in China, more than two thirds of consumers in urban areas have heard of GM foods. But their knowledge on biotechnology was limited. Chinese consumers' acceptance of and willingness to buy GM foods was much higher than in other countries. Chinese consumers also demonstrated great variance in their acceptance of different GM foods. Information and prices of GM foods were two important factors affecting consumers' attitudes toward GM foods. Based on the findings of this study and given that our sample is in the more developed eastern Urban China, we conclude that the commercialization of GM foods is not likely to receive great resistance from the consumers in China. PMID- 16469415 TI - Investigation on recombinant hirudin via oral route. AB - The possibility for oral administration of peptide recombinant hirudin variant (rHV2-K47) as an anticoagulant agent was evaluated in several aspects. The proteolytic properties of rHV2-K47 and its stability during storage were examined by in vitro experiments. Radiolabeled rHV2-K47 was infused into the duodenum of rats and rHV2-K47 absorbed into serum was shown to be intact by electrophoresis pattern. The in vivo coagulation time of blood from mouse was prolonged significantly after oral administration of rHV2-K47. The bioavailability (F) of rHV2-K47 via oral route reached 10.11% in comparison with intravenous administration as gold standard. All the results suggested that rHV2-K47 could be delivered successfully via the oral route. PMID- 16469416 TI - Nitric oxide-evoked glutamate release and cGMP production in cerebellar slices: control by presynaptic 5-HT1D receptors. AB - We previously reported that pre- and postsynaptic 5-hydroxytryptamine (5-HT) receptors effectively control glutamatergic transmission in adult rat cerebellum. To investigate where 5-HT acts in the glutamate ionotropic receptors/nitric oxide/guanosine 3',5'-cyclic monophosphate (cGMP) pathway, in the present study 5 HT modulation of the cGMP response to the nitric oxide donor S-nitroso penicillamine (SNAP) was studied in adult rat cerebellar slices. While cGMP elevation produced by high-micromolar SNAP was insensitive to 5-HT, 1 microM SNAP, expected to release nitric oxide in the low-nanomolar concentration range, elicited cGMP production and endogenous glutamate release both of which could be prevented by activating presynaptic 5-HT1D receptors. Released nitric oxide appeared responsible for cGMP production and glutamate release evoked by 1 microM SNAP, as both the effects were mimicked by the structurally unrelated nitric oxide donor 2-(N,N-diethylamino)-diazenolate-2-oxide (0.1 microM). Dependency of the 1 microM SNAP-evoked release of glutamate on external Ca2+, sensitivity to presynaptic release-regulating receptors and dependency on ionotropic glutamate receptor functioning, suggest that nitric oxide stimulates exocytotic-like, activity-dependent glutamate release. Activation of ionotropic glutamate receptors/nitric oxide synthase/guanylyl cyclase pathway by endogenously released glutamate was involved in the cGMP response to 1 microM SNAP, as blockade of NMDA/non-NMDA receptors, nitric oxide synthase or guanylyl cyclase, abolished the cGMP response. To conclude, in adult rat cerebellar slices low-nanomolar exogenous nitric oxide could facilitate glutamate exocytotic-like release possibly from parallel fibers that subsequently activated the glutamate ionotropic receptors/nitric oxide/cGMP pathway. Presynaptic 5-HT1D receptors could regulate the nitric oxide-evoked release of glutamate and subsequent cGMP production. PMID- 16469417 TI - The neural cell adhesion molecule NCAM regulates neuritogenesis by multiple mechanisms of interaction. AB - The neural cell adhesion molecule NCAM and its glycosylation with polysialic acid (polySia) are crucially involved in proliferation, migration and differentiation of neural progenitors. Modification with polySia, homophilic and heterophilic interactions set the function of NCAM, but little is known on their interplay. We have shown recently that removal of polySia induces neuronal differentiation via heterophilic NCAM interactions at cell contacts between SH-SY5Y neuroblastoma cells. Here we analyze the additional impact of NCAM-positive fibroblasts as a ligand-presenting cellular environment, a model often used to demonstrate the neuritogenic effect of homophilic NCAM interactions. Native SH-SY5Y cells did not respond to interactions with fibroblast NCAM. However, after induction of neuronal differentiation by retinoic acid the previously ineffective NCAM signals activated extracellular signal-regulated kinase (ERK) and promoted neuritogenesis. Removal of polySia increased neuritogenesis in retinoic acid treated cells additive to the NCAM substrate effect. The change in responsiveness to substrate NCAM was associated with a rearrangement of polysialylated NCAM away from its enrichment at homotypic cell-cell contacts and with the appearance of non-polysialylated NCAM, i.e. changes facilitating NCAM interactions with the substrate. Thus, heterophilic and homophilic NCAM interactions are integrated into the cell's response yet they have the capacity to independently trigger neuritogenesis. The actual occurrence of each of these interactions, however, depends on the cellular context, targeted cell surface presentation of NCAM and the dynamic regulation of its modification by polysialic acid. In summary, this study reveals how the complex interplay of NCAM interactions and polysialylation provides an elaborate system to regulate neuritogenesis. PMID- 16469418 TI - Systemic hypoperfusion is associated with executive dysfunction in geriatric cardiac patients. AB - The present study examines the relationship between systemic hypoperfusion via cardiac output (CO) and neuropsychological performances emphasizing executive function in an aging cohort. Geriatric outpatients with treated, stable cardiovascular disease (CVD) and no history of neurological illness (n=72, ages 56-85) were administered cognitive measures with an emphasis on executive functioning. Echocardiogram findings were used to stratify participants into two groups: low CO (<4.0 L/min) and normal CO (> o r=4.0 L/min). Between-group comparisons were made using ANCOVAs adjusting for systolic blood pressure. The low CO group performed significantly worse than the normal CO group on DKEFS Tower Test and DKEFS Trail Making Test. No significant between-group differences were noted for any of the other cognitive indices. Findings suggest that reduced CO is associated with poorer executive functioning among geriatric outpatients with stable CVD, as the cognitive profile emphasizes a relationship between systemic hypoperfusion and problems with sequencing and planning. The executive dysfunction profile may be secondary to reduced blood flow to vulnerable subcortical structures implicated in frontal-subcortical circuitry. PMID- 16469419 TI - Structural MRI covariance patterns associated with normal aging and neuropsychological functioning. AB - Structural magnetic resonance imaging (MRI) studies have shown dramatic age associated changes in grey and white matter volume, but typically use univariate analyses that do not explicitly test the interrelationship among brain regions. The current study used a multivariate approach to identify covariance patterns of grey and white matter tissue density to distinguish older from younger adults. A second aim was to examine whether the expression of the age-associated covariance topographies is related to performance on cognitive tests affected by normal aging. Eighty-four young (mean age=24.0) and 29 older (mean age=73.1) participants were scanned with a 1.5T MRI machine and assessed with a cognitive battery. Images were spatially normalized and segmented to produce grey and white matter density maps. A multivariate technique, based on the subprofile scaling model, was used to capture sources of between- and within-group variation to produce a linear combination of principal components that represented a "pattern" or "network" that best discriminated between the two age groups. Univariate analyses were also conducted with statistical parametric maps. Grey and white matter covariance patterns were identified that reliably discriminated between the groups with greater than 0.90 sensitivity and specificity. The identified patterns were similar for the univariate and multivariate techniques, and involved widespread regions of the cortex and subcortex. Age and the expression of both patterns were significantly associated with performance on tests of attention, language, memory, and executive functioning. The results suggest that identifiable networks of grey and white matter regions systematically decline with age and that pattern expression is linked to age-related cognitive decline. PMID- 16469420 TI - Peptide-binding assays and HLA II transgenic Abeta degrees mice are consistent and complementary tools for identifying HLA II-restricted peptides. AB - The identification of MHC class II-restricted peptides has become a priority for the development of peptide-based prophylactic and therapeutic vaccines. The aim of this study was to assess the correlations between peptide-binding assays on purified HLA II molecules and immunization of human HLA II transgenic mice deficient in murine class II molecules (Abeta degrees ). We used as models two MHC class II-restricted peptides, one derived from the HIV Nef regulatory protein (Nef (56-68)) and the other from the Schistosoma mansoni 28-kDa glutathione-S transferase (Sm28GST (190-211)). High correlations were found between the two approaches, which showed that the Nef (56-68) and Sm28GST (190-211) peptides may represent promiscuous ligands for HLA-DQ and for HLA-DR molecules, respectively. We suggest a rational method based on the combination of peptide-binding assays and HLA II transgenic mice experiments as consistent and complementary tools for selecting T helper epitopes. PMID- 16469421 TI - High-resolution alpha spectrometry under field conditions - fast identification of alpha particle emitting radionuclides from air samples. AB - Alpha particle emitting radionuclides may be released into the atmosphere as a result of a severe nuclear accident, the malevolent use of radioactive material or some other incident. Rapid detection methods are then of crucial importance for carrying out well-timed protective actions. The present paper demonstrates that a method in which an aerosol sample is collected on a filter and directly analysed by high-resolution alpha spectrometry in a vacuum chamber, but without radiochemical sample treatment, is suitable for field measurements. A detection limit of less than 0.1 Bqm(-3) for (239)Pu is achieved assuming 1-h sampling using a glass-fibre filter and 1-h data acquisition. This value is higher than in the case when sampling is performed using a porous membrane filter. A measurement system consisting of standard components is mounted in a vehicle designed for rapid field measurements. PMID- 16469422 TI - Long-term efficiency and stability of wetlands for treating wastewater of a lead/zinc mine and the concurrent ecosystem development. AB - A constructed wetland system in Guangdong Province, South of China has been used for treating Pb/Zn mine discharge since 1985. The performance in the purification of the mine discharge and the concurrent ecosystem development within the system during the period of 1985-2000 has been studied. The untreated wastewater contained rather high concentrations of cadmium (Cd) (0.05 mg L(-1)), lead (Pb) (11.5 mg L(-1)), and zinc (Zn) (14.5 mg L(-1)), which greatly exceed the upper limits for industrial wastewater discharge in China. The constructed wetland system effectively removed Cd by 94.00%, Pb by 99.04%, Zn by 97.30%, and total suspended solids (TSS) by 98.95% from the mine discharge over a long period (over 16 years) leading to significant improvement in water quality; it was also found that there were no significantly annual or monthly variations in pH values, As, Cd, Hg, Pb, and Zn concentrations in water collected from the outlet of the wetland. Moreover, diversity and abundance of living organisms, including protozoan, higher plants, terrestrial animals, and birds, increased gradually. The 16-year monitoring results showed a reciprocal relationship, at a certain extent, between restoration of the wetland ecosystem, in other words, the maturity of the wetland, and the long-term efficiency and stability on purifying heavy metal-contaminated wastewater. PMID- 16469423 TI - Heptachlor epoxide induces a non-capacitative type of Ca2+ entry and immediate early gene expression in mouse hepatoma cells. AB - The effects of the organochlorine (OC) liver tumor promoter heptachlor epoxide (HE) and a related non-tumor promoting OC, delta-hexachlorocyclohexane (delta HCH), on the dynamics of intracellular calcium (Ca2+) were investigated in mouse 1c1c7 hepatoma cells. HE induced a non-capacitative, Ca2+ entry-like phenomenon, which was transient and concentration-dependent with 10 and 50 microM HE. The plasma membrane Ca2+ channel blocker SKF-96365 antagonized this HE-induced Ca2+ entry. delta-HCH failed to induce Ca2+ entry, rather it antagonized the HE induced Ca2+ entry. Both HE and delta-HCH induced Ca2+ release from endoplasmic reticulum (ER) at treatment concentrations as low as 10 microM; at 50 microM, the former induced 5x as much Ca2+ release as the latter. The HE-induced Ca2+ release from the ER was antagonized using the IP3 receptor/channel blocker xestospongin C, suggesting that HE induces ER Ca2+ release through the IP3 receptor/channel pore. These results show that the effect of HE on cellular Ca2+ mimics that of mitogens such as epidermal and hepatocyte growth factors. They also provide insight into the similarities and differences between tumorigenic and non tumorigenic OCs, in terms of the mechanisms and the extent of the [Ca2+]i increased by these agents. PMID- 16469424 TI - European regulations on nutraceuticals, dietary supplements and functional foods: a framework based on safety. AB - This article describes the legislation that is relevant in the marketing of functional foods in the European Union (EU), how this legislation was developed as well as some practical consequences for manufacturers, marketers and consumers. It also addresses some concrete examples of how the EU's safety requirements for food products have impacted a range of product categories. In the late nineties, research into functional ingredients was showing promising prospects for the use of such ingredients in foodstuffs. Due mainly to safety concerns, these new scientific developments were accompanied by an urgent call for legislation. The European Commission 2000 White Paper on Food Safety announced some 80 proposals for new and improved legislation in this field. Among others, it foresaw the establishment of a General Food Law Regulation, laying down the principles of food law and the creation of an independent Food Authority endowed with the task of giving scientific advice on issues based upon scientific risk assessment with clearly separated responsibilities for risk assessment, risk management and risk communication. Since then, more than 90% of the White Paper proposals have been implemented. However, there is not, as such, a regulatory framework for 'functional foods' or 'nutraceuticals' in EU Food Law. The rules to be applied are numerous and depend on the nature of the foodstuff. The rules of the general food law Regulation are applicable to all foods. In addition, legislation on dietetic foods, on food supplements or on novel foods may also be applicable to functional foods depending on the nature of the product and on their use. Finally, the two proposals on nutrition and health claims and on the addition of vitamins and minerals and other substances to foods, which are currently in the legislative process, will also be an important factor in the future marketing of 'nutraceuticals' in Europe. The cornerstone of EU legislation on food products, including functional foods and nutraceuticals is 'safety'. Decisions on the safety-basis of legislation are based on risk analysis, in which scientific risk assessment is performed by the European Food Safety Authority and risk management is performed by the European Commission, the Member States, and in case of legislation, together with the European Parliament. In the risk management phase, both the precautionary principle and other legitimate factors may be considered in choosing the best way of dealing with an issue. Due to the numerous pieces of legislation applying and to the different procedures to be followed, the process of having 'functional foods' ready for the market is certainly a costly and time-consuming task. However, it may also be clearly worth it in terms of market success and improved consumer health. PMID- 16469425 TI - The impact of cGMP compliance on consumer confidence in dietary supplement products. AB - The FDA estimates that US citizens spend more than $ 8.5 billion a year on dietary supplements and world wide the market is estimated at more than $ 60 billion. However, although a majority of consumers express confidence in the safety of these products, 74% believe the government should be more involved in ensuring that these products are safe and efficacious. Recent regulatory initiatives such as the imminent adoption of cGMPs for dietary supplements in the US, implementation of cGMPs in Canada and the recent EU dietary supplement initiative represent legislative and industry response to public clamor for more comprehensive oversight of dietary supplements. Regardless of mandated practices, the majority of dietary supplement manufacturers have done an excellent job of protecting the safety and quality of their products. The promulgation of these cGMPs will help ensure consumers that equal standards are followed throughout the industry. For some companies with established processes based on existing food or pharmaceutical cGMP regulations, the transition will be relatively painless while, for many, it will represent a significant increase in the level of documentation and testing. However, consumers deserve and demand that products meet standards for safety and quality and the implementation of cGMPs for these products are an important first step. Although the cGMPs are designed to ensure products are safe from a standpoint of identity, purity, quality, strength and composition, they do not address preclinical or clinical testing of ingredients for safety or efficacy. This would involve ingredients meeting the requirements of Generally Recognized as Safe (GRAS) status or going through the New Dietary Ingredient (NDI) process. PMID- 16469426 TI - The full-length cDNA of anticoagulant protein infestin revealed a novel releasable Kazal domain, a neutrophil elastase inhibitor lacking anticoagulant activity. AB - Infestins are Kazal-type serine proteinase inhibitors found in the midgut of the Chagas' disease vector, Triatoma infestans. In previous studies, we characterized two double-headed infestins with potent anticoagulant activity; infestin 1-2, which inhibits thrombin and infestin 3-4, a factor XIIa inhibitor. In the present work, we have cloned the full-length cDNA of infestins' precursor. The translated cDNA predicted a polypeptide containing a signal peptide and seven Kazal-type domains, four domains from infestin 1-2 and infestin 3-4, and three new domains. Northern blot analysis confirmed that infestins are synthesized in a single transcript (approximately 1,800 bp) in the insect midgut, but not in salivary glands. Based on the cDNA sequence, the three new Kazal domains were named infestin 1R, 2R and 3R. Infestin 2R-3R has 77% amino acid sequence identity to infestin 1-2 and the same basic amino acid residue at P1 position in the inhibitory reactive site suggesting that these two proteins have a similar inhibitory specificity. In contrast, infestin 1R has two different characteristics when compared to the other infestins: i) a hydrophobic amino acid residue at P1 position in the inhibitory reactive site and ii) a prediction to be processed as a single Kazal domain. These two characteristics were experimentally demonstrated by the purification of native infestin 1R from T. infestans midgut. Native infestin 1R was shown to be processed as a single Kazal domain by mass spectrometry and it was able to inhibit neutrophil elastase, subtilisin A and chymotrypsin. To further characterize infestin 1R inhibitory activity, it was expressed as a recombinant protein in bacteria. Recombinant infestin 1R inhibited neutrophil elastase with the same K(i) of the native inhibitor. Moreover, it inhibited subtilisin A, chymotrypsin and proteinase K but did inhibit neither thrombin nor coagulation assays. In conclusion, unlike the other described infestins, infestin 1R did not present anticoagulant activity and is processed as a single Kazal domain with inhibitory specificity towards proteases that hydrolyze peptide bonds after hydrophobic amino acid residues. PMID- 16469427 TI - Site-directed mutagenesis study of the microenvironment characteristics of Lys213 of Saccharomyces cerevisiae phosphoenolpyruvate carboxykinase. AB - Saccharomyces cerevisiae phosphoenolpyruvate (PEP) carboxykinase catalyzes the reversible formation of oxaloacetate and adenosine triphosphate from PEP, adenosine diphosphate and carbon dioxide, and uses Mn(2+) as the activating metal ion. Comparison with the crystalline structure of homologous Escherichia coli PEP carboxykinase [Tari et al. Nature Struct. Biol. 4 (1997) 990-994] shows that Lys(213) is one of the ligands to Mn(2+) at the enzyme active site. Coordination of Mn(2+) to a lysyl residue is infrequent and suggests a low pK(a) value for the epsilon-NH(2) group of Lys(213). In this work, we evaluate the role of neighboring Phe(416) in contributing to provide a low polarity microenvironment suitable to keep the epsilon-NH(2) of Lys(213) in the unprotonated form. Mutation Phe416Tyr shows that the introduction of a hydroxyl group in the lateral chain of the residue produces a substantial loss in the enzyme affinity for Mn(2+), suggesting an increase of the pK(a) of Lys(213). A study of the effect of pH on K(m) for Mn(2+) indicate that the affinity of recombinant wild type enzyme for the metal ion is dependent on deprotonation of a group with pK(a) of 7.1+/-0.2, compatible with the low pK(a) expected for Lys(213). This pK(a) value increases at least 1.5 pH units upon Phe416Tyr mutation, in agreement with the expected effect of an increase in the polarity of Lys(213) microenvironment. Theoretical calculations of the pK(a) of Lys(213) indicate a value of 6.5+/-0.9, and it increases to 8.2+/-1.6 upon Phe416Tyr mutation. Additionally, mutation Phe416Tyr causes a loss of 1.3 kcal mol(-1) in the affinity of the enzyme for PEP, an effect perhaps related to the close proximity of Phe(416) to Arg(70), a residue previously shown to be important for PEP binding. PMID- 16469429 TI - Complications of transurethral resection of the prostate (TURP)--incidence, management, and prevention. AB - OBJECTIVES: To update the complications of transurethral resection of the prostate (TURP), including management and prevention based on technological evolution. METHODS: Based on a MEDLINE search from 1989 to 2005, the 2003 results of quality management of Baden-Wurttemberg, and long-term personal experience at three German centers, the incidence of complications after TURP was analyzed for three subsequent periods: early (1979-1994); intermediate (1994-1999); and recent (2000-2005) with recommendations for management and prevention. RESULTS: Technological improvements such as microprocessor-controlled units, better armamentarium such as video TUR, and training helped to reduce perioperative complications (recent vs. early) such as transfusion rate (0.4% vs. 7.1%), TUR syndrome (0.0% vs. 1.1%), clot retention (2% vs. 5%), and urinary tract infection (1.7% vs. 8.2%). Urinary retention (3% vs. 9%) is generally attributed to primary detrusor failure rather than to incomplete resection. Early urge incontinence occurs in up to 30-40% of patients; however, late iatrogenic stress incontinence is rare (<0.5%). Despite an increasing age (55% of patients are older than 70), the associated morbidity of TURP maintained at a low level (<1%) with a mortality rate of 0-0.25%. The major late complications are urethral strictures (2.2-9.8%) and bladder neck contractures (0.3-9.2%). The retreatment rate range is 3-14.5% after five years. CONCLUSIONS: TURP still represents the gold standard for managing benign prostatic hyperplasia with decreasing complication rates. Technological alternatives such as bipolar and laser treatments may further minimize the risks of this technically difficult procedure. PMID- 16469430 TI - Testicular zinc finger protein recruits histone deacetylase 2 and suppresses the transactivation function and intranuclear foci formation of agonist-bound androgen receptor competitively with TIF2. AB - We previously reported that testicular zinc finger protein (TZF) is a corepressor for androgen receptor (AR). The present study demonstrated that a central portion (amino acids 512-663) of TZF, TZF(512-663), is responsible for both binding to AR and repressing the transactivation. TZF recruited endogenous histone deacetylase 2 (HDAC2) and formed a complex with agonist-bound AR. Imaging analyses showed that TZF and TZF(512-663) were recruited by AR and simultaneously impaired distinct AR foci formation. Quantification of the foci number using a three dimensional imaging method revealed that the number of intranuclear AR foci was related to its transactivation activity. Moreover, increased levels of TZF dissociated a coactivator, TIF2, from the AR foci and vice versa. These results indicate that the ligand-dependent transactivation function of AR is quantitatively related to its intranuclear foci formation, and suggest that corepressors, such as TZF, act on these intranuclear events competitively with coactivators. PMID- 16469431 TI - Differential regulation of cell growth and gene expression by FGF-2 and FGF-4 in pituitary lactotroph GH4 cells. AB - Fibroblast growth factors, FGF-2 and FGF-4, are reported to play divergent roles in pituitary differentiation and tumor formation, stimulating cell differentiation or proliferation, respectively. However, mitogenic responses to FGFs have not been extensively characterized and little is known about the molecular mechanisms by which specific FGF isoforms may mediate distinct biological responses. Here we show that FGF-4 but not FGF-2 stimulated DNA synthesis and cell proliferation in GH4 cells. Microarray analyses revealed that FGF-4 induced expression of several oncogenes, growth factor receptors and cell cycle control proteins (e.g. cyclin D3/cdk4, N-myc, c-Raf, insulin and thyroid hormone receptors) while FGF-2 had no effect or down regulated these same genes. These transcriptional responses are consistent with a proliferative and/or tumorigenic role for FGF-4 versus a growth inhibitory effect of FGF-2. FGF-2 and FGF-4 also differentially regulated MAP kinase phosphorylation, which may underlie their isoform-specific effects on cell growth and gene expression. PMID- 16469432 TI - Id-1 overexpression in invasive ductal carcinoma cells is significantly associated with intratumoral microvessel density in ER-negative/node-positive breast cancer. AB - The aim of this study is to investigate the possible role of inhibitor of DNA binding (Id-1) overexpression in human breast cancer. We examined Id-1 expression by immunohistochemistry in 263 human breast cancers, 15 in situ lesions and 248 invasive cancers to investigate the relationship between its expression and various clinicopathological factors. Id-1 expression was significantly higher in invasive ductal carcinoma than in in situ ductal carcinoma or other invasive cancer subtypes (P=0.029 and 0.006, respectively). We also examined the association between Id-1 expression and tumor angiogenesis by measuring microvessel densities (MVD). Regarding the endothelial cells of microvessels showed negative or very weak Id-1 expression, Id-1 overexpression was found to be significantly related to MVD (P=0.014). Furthermore, Id-1 overexpression was found to be significantly associated with higher MVD in the ER-negative and node involved subgroups of breast cancer (P=0.040 and 0.046, respectively). These data indicate that Id-1 overexpression is significantly associated with tumor angiogenesis, especially in the ER-negative and node-positive subtypes of invasive breast cancer. Thus, Id-1 presents a possible therapeutic antitumor target molecule in ER-negative and node-positive breast cancer. PMID- 16469433 TI - Sprouty and cancer: the first terms report. AB - The Ras/Erk signaling pathway has a central role in development of multi-cellular organisms as well as in signal transmission in the mature individual. Recently, a family of genes, designated Sprouty, induced by the Ras/Erk pathway was found to specify proteins that inhibited the upstream pathway. Being in a position that is likely to control well-characterized oncogene products suggested that the expression levels of the Sprouty genes may be relevant in human carcinogenesis. Early data on the deregulation of Sprouty expression in breast, prostate and liver cancers is discussed along with the notion that some of them might have potential as tumour markers or that the derived proteins may act as tumour suppressors. PMID- 16469434 TI - Red soil as a regenerable sorbent for high temperature removal of hydrogen sulfide from coal gas. AB - In this study, hydrogen sulfide (H(2)S) was removed from coal gas by red soil under high temperature in a fixed-bed reactor. Red soil powders were collected from the northern, center and southern of Taiwan. They were characterized by XRPD, porosity analysis and DCB chemical analysis. Results show that the greater sulfur content of LP red soils is attributed to the higher free iron oxides and suitable sulfidation temperature is around 773K. High temperature has a negative effect for use red soil as a desulfurization sorbent due to thermodynamic limitation in a reduction atmosphere. During 10 cycles of regeneration, after the first cycle the red soil remained stable with a breakthrough time between 31 and 36 min. Hydrogen adversely affects sulfidation reaction, whereas CO exhibits a positive effect due to a water-shift reaction. COS was formed during the sulfidation stage and this was attributed to the reaction of H(2)S and CO. Results of XRPD indicated that, hematite is the dominant active species in fresh red soil and iron sulfide (FeS) is a product of the reaction between hematite and hydrogen sulfide in red soils. The spinel phase FeAl(2)O(4) was found during regeneration, moreover, the amount of free iron oxides decreased after regeneration indicating the some of the free iron oxide formed a spinel phase, further reducting the overall desulfurization efficiency. PMID- 16469435 TI - Studies of removal of platinum(IV) ion microquantities from the model solutions of aluminium, copper, iron, nickel and zinc chloride macroquantities on the anion exchanger Duolite S 37. AB - Platinum has been widely applied in catalytic industry and the recovery of noble metals from industrial wastes becomes an economic issue. The laboratory studies of platinum(IV) microquantities removal from 1M aluminium, copper, iron, nickel and zinc chloride solutions in 0.1M hydrochloric acid solutions on the anion exchanger Duolite S 37 of the functional secondary and tertiary amine groups were carried out. For this anion exchanger the fraction extracted values (%E, Pt(IV)) as well as the sorption isotherms were determined depending on the kind of aqueous phase and phase contact time. Moreover, the bed and weight distribution coefficients as well as working and total ion-exchange capacities were calculated from the platinum(IV) breakthrough curves. Kinetic parameters were determined. PMID- 16469436 TI - Analysis of n-alkanes in water samples by means of headspace solvent microextraction and gas chromatography. AB - A simple and efficient headspace solvent microextraction (HSME) was developed for the simultaneous determination of the trace concentrations of some n-alkanes in water samples. Therefore, a microdrop of an organic solvent was extruded from the needle tip of a gas chromatographic syringe to the headspace above the surface of the solution in a sealed vial. Then the volatile organic compounds are extracted and concentrated in the microdrop. Next, the microdrop was retracted into the microsyringe and injected directly into the gas chromatograph. Experimental parameters which control the performance of HSME such as the type of microextraction solvent, organic drop and sample volume, sample stirring rate, sample solution and microsyringe needle temperatures, salt addition and exposure time profiles were investigated and optimized. Finally, the enrichment factor, dynamic linear range (DLR), limit of detection (LOD) and precision of the method were evaluated. Using optimum extraction conditions, good linearity with correlation coefficients in the range of 0.995LUV>SUV. An opposite behaviour was observed for FATMLV, probably due to the freezing phase required by such a preparation method, which reduced the complex solubility. Moreover, it was not possible to use higher complex concentrations, due to the destabilizing effect of cyclodextrins toward the liposomal membrane. Permeability studies of drug-HPbetaCyd complexes, directly in solution or incorporated in liposomes, performed across artificial membranes simulating the skin behaviour, highlighted, as expected, a prolonged release effect of liposomal formulations. Furthermore, the drug permeation rate depended on the vesicle characteristics and varied in the order: SUV>MLV=FATMLV>LUV. Therefore, the most suitable liposome preparation method can be suitably selected on the basis of drug encapsulation efficiency and/or desired drug release rate. PMID- 16469461 TI - The dorsal cochlear nucleus as a participant in the auditory, attentional and emotional components of tinnitus. AB - The dorsal cochlear nucleus (DCN) has been modeled in numerous studies as a possible source of tinnitus-generating signals. This hypothesis was originally developed on the basis of evidence that the DCN becomes hyperactive following exposure to intense noise. Since these early observations, evidence that the DCN is an important contributor to tinnitus has grown considerably. In this paper, the available evidence to date will be summarized. In addition, the DCN hypothesis of tinnitus can now be expanded to include possible involvement in other, non-auditory components of tinnitus. It will be shown by way of literature review that the DCN has direct connections with non-auditory brainstem structures, such as the locus coeruleus, reticular formation and raphe nuclei, that are implicated in the control of attention and emotional responses. The hypothesis will be presented that attentional and emotional disorders, such as anxiety and depression, which are commonly associated with tinnitus, may result from an interplay between these non-auditory brainstem structures and the DCN. Implicit in this hypothesis is that attempts to develop effective anti-tinnitus therapies are likely to benefit from a greater understanding of how the levels of activity in the DCN are influenced by different states of activation of these non auditory brainstem structures and vice versa. PMID- 16469462 TI - Effect of Cardiospermum halicacabum on ethanol-induced gastric ulcers in rats. AB - Ethanol extract of Cardiospermum halicacabum Linn. (Sapindaceae), in a concentration dependant manner (200-600mg/kg) inhibited gastric ulcers induced by oral administration of absolute ethanol. Further, the extract administration to rats resulted in an increase in levels of gastric glutathione and a decrease in alkaline phosphatase activity. The extract also exhibited potent in vitro hydroxyl radical scavenging and inhibition of lipid peroxidation activities. The extract was found to be devoid of any conspicuous acute and short-term toxicity in rats. PMID- 16469463 TI - Antiviral activities of purified compounds from Youngia japonica (L.) DC (Asteraceae, Compositae). AB - The ethanol extract of a biannual medicinal herb, Youngia japonica (commonly known as Oriental hawk's beard) was reported previously to have potent antiviral activity against respiratory syncytial virus (RSV) cultured in HEp-2 cells. Three anti-microbial agents, namely 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, and luteolin-7-O-glucoside were subsequently purified and chemically characterized from the ethanol extract of Youngia japonica. The two dicaffeoylquinic acids exhibited prominent anti-RSV with 50% inhibitory concentration (IC50) of 0.5 microg/ml in vitro. Luteolin-7-O-glucoside together with the two dicaffeoylquinic acids were also manifested to have some antibacterial activity towards the causal agents of food-borne disease, namely Vibrio cholerae and Vibrio parahaemolyticus at the concentration of 2mg/ml. Bacillus cereus was sensitive to 3,4-dicaffeoylquinic acid and 3,5 dicaffeoylquinic acid only, but not to luteolin-7-O-glucoside. PMID- 16469464 TI - Evaluation of thoracic abnormalities on 64-row multi-detector row CT: comparison between axial images versus coronal reformations. AB - PURPOSE: To evaluate the capability of coronal reformations of chest on 64-row MDCT in demonstrating thoracic abnormalities in comparison with axial images. MATERIALS AND METHODS: Thirty-eight consecutive patients who underwent pulmonary CTA on 64-row MDCT were retrospectively studied with institutional review board (IRB) approval. Contiguous 2 mm axial and coronal images were reviewed independently with a 1-week interval, by consensus reading of two board-certified radiologists. Overall image quality was graded using a five-point scale. Abnormalities in mediastinum, hilum, pulmonary vessels, aorta, heart, esophagus, pleura, chest wall, and lung parenchyma were scored: 1 = definitely absent, 2 = probably absent, 3 = equivocal, 4 = probably present, 5 = definitely present. Scores on axial and coronal images were compared using weighted kappa analysis. RESULTS: Overall image quality was not different with statistical relevance between axial and coronal images (mean/median scores; 3.7/4; 3.6/4, respectively, P = 0.286, Wilcoxon signed-rank test). Significant agreement was observed between axial and coronal scores (mean weighted kappa, 0.661; range, 0.362-1). Agreement was almost perfect for pneumothorax, lung and pleural mass, effusion and consolidation (weighted kappa=0.833-1); substantial for pulmonary embolism, trachea, mediastinal lymphadenopathy and non-skeletal chest wall lesion, heart, esophagus, and emphysema (weighted kappa, 0.618-0.799); moderate for atelectasis, mediastinum, hilar nodes, aorta, other lung lesions, skeletal chest wall lesions, linear scarring, nodules > 1 cm, pulmonary artery abnormalities and pleural thickening (weighted kappa, 0.405-0.592); and fair for nodules < 1 cm (weighted kappa = 0.362). CONCLUSION: Coronal reformations on 64-row MDCT had substantial agreement with axial images for evaluation of the majority of thoracic abnormalities. PMID- 16469465 TI - Osmoprotection of Salmonella enterica serovar Typhimurium by Ngamma acetyldiaminobutyrate, the precursor of the compatible solute ectoine. AB - N(gamma)-acetyl-2,4-diaminobutyrate (NADA), the precursor of the compatible solute ectoine, was shown to function as an osmoprotectant for the non-halophilic bacterium Salmonella enterica serovar Typhimurium. The addition of NADA containing extracts of an ectoine synthase mutant of the broad salt-growing halophile Chromohalobacter salexigens DSM 3043(T) could alleviate the inhibitory effects of high salinity in S. enterica, which lacks the ectoine biosynthetic pathway. NADA, purified from extracts of the mutant, protected S. enterica against salinity stress. This osmoprotective effect was slightly lower than that of ectoine, but more potent than that of hydroxyectoine. Accumulation of purified NADA by S. enterica was demonstrated by (13)C-NMR spectroscopy and HPLC analysis. In addition, it was shown that NADA was taken up by S. enterica via the ProP and ProU transport systems, which are known to transport glycine betaine and proline. This finding provides evidence that these permeases can recognize a diaminoacid that carries an unsubstituted alpha-amino group. This is the first time that NADA has been connected with osmoprotective functions in non-halophilic bacteria. PMID- 16469466 TI - Analysis of lecithin-cholesterol mixtures using Raman spectroscopy. AB - FT-Raman spectroscopy has been used to investigate interactions between lecithin and cholesterol. Raman spectra of lecithin show multiple peaks which can be classified into three regions: hydrophobic chain, interfacial, and headgroup regions. Binary lipid mixtures (1:1, w/w, lecithin:cholesterol) were prepared by physical mixing, granulation, coprecipitation, hydration and heating (65 degrees C), and heating (120 degrees C). Regardless of the preparation method, no changes in the spectra were observed in the hydrophobic region. A shift in the wavenumber of the choline methyl asymmetric stretching mode was observed when the samples were prepared by coprecipitation, hydration and heating (65 degrees C), and heating (120 degrees C). This may indicate a modification of phospholipids in the headgroup region in these samples. The difference in degrees of frequency shift (physical mixing approximately granulationor=96%) and uniform (purity >or=97%). After serum- as well as LTB-4-induced transmigration granulocytes produced the same amounts of reactive oxygen species (ROS) as non-migrated cells in HK cell suspension. Their morphology, staining characteristics and flow cytometric scatter characteristics, plus their ability to produce ROS characterised the transmigrated granulocytes as neutrophils. The quantitative transmigration system described here could also serve as an excellent tool for the selective attraction and isolation of highly purified carp neutrophils from HK cell suspensions. PMID- 16469506 TI - Role of each residue in catalysis in the active site of pyrimidine nucleoside phosphorylase from Bacillus subtilis: a hybrid QM/MM study. AB - Pyrimidine nucleoside phosphorylase (PYNP) catalyzes the reversible phosphorolysis of pyrimidines in the nucleotide synthesis salvage pathway. We have built a model of a closed active conformation of the three-dimensional structure of PYNP from Bacillus subtilis. Using docking, molecular dynamics, and hybrid quantum-mechanical/molecular-mechanical methods to study the reaction mechanics between PYNP and a substrate, we identified the role of each residue in the active site during the entire catalytic process. The results indicate that the function of His(82), Arg(169), and Lys(188) is to stabilize the uridine in a high-energy conformation by means of electrostatic interactions and that these residues are involved in catalysis. In addition, the function of Asp(162) is likely to activate Lys(188) for phosphorolytic catalysis through polarization effects. PMID- 16469508 TI - Gamma-band activity dissociates between matching and nonmatching stimulus pairs in an auditory delayed matching-to-sample task. AB - Electro- and magnetoencephalography studies have suggested that increased gamma band activity (GBA) is a correlate of activated neural stimulus representations. In this study, a delayed matching-to-sample paradigm for auditory spatial information was employed to investigate the role of magnetoencephalographic gamma band activity in the differentiation between matching and nonmatching stimulus pairs. Twelve subjects made same-different judgments about the lateralization angle of pairs of filtered noise stimuli (S1 and S2) presented with 0.8-s delays. One half of the subjects had to respond to matching stimulus pairs, the other half to nonmatching stimulus pairs. Cortical oscillatory activity in the memory task was compared to a control task requiring the detection of background noise intensity changes. Memory-related GBA increases were revealed over midline parietal areas in the middle of the delay phase and during the presentation of S2 and over frontocentral areas at the end of the delay phase. This replicated previous findings. In addition, nonmatching trials were associated with increased GBA over right parietal areas in response to S2. The midline parietal GBA increase during S2 in the memory condition may have reflected the representation of S1 needed for a comparison between S1 and S2. When S1 and S2 were identical, no further representation was required. In contrast, for nonmatching pairs, a second representation was activated over right parietal areas. PMID- 16469510 TI - Measuring dopamine neuromodulation in the thalamus: using [F-18]fallypride PET to study dopamine release during a spatial attention task. AB - We used the highly selective D2/D3 dopamine PET radioligand [F-18]fallypride to demonstrate that cognitive task induced dopamine release can be measured in the extrastriatal region of the thalamus, a region containing 10-fold fewer D2 dopamine receptors than the striatum. Human studies were acquired on 8 healthy volunteers using a single [F-18]fallypride injection PET imaging session. A spatial attention task, previously demonstrated to increase FDG uptake in the thalamus, was initiated following a period of radioligand uptake. Thalamic dopamine release was statistically tested by measuring time-dependent alterations in the kinetics (focusing on specific binding) of the [F-18]fallypride using the linearized extension of the simplified reference region model. Voxel-based analysis of the dynamic PET data sets revealed a high correlation (r = 0.86, P = 0.0067) between spatial attention task performance and thalamic dopamine release. Various aspects of the kinetic model were analyzed to address concerns such as blood flow artifacts and model bias, as well as issues with task timing and regional variations in D2/D3 receptor density. In addition to the thalamus, measurement of dopamine neuromodulation using [F-18]fallypride and a single injection PET protocol can be extended to other extrastriatal regions of the brain, such as the amygdala, hippocampus, and regions of the temporal cortex. However, issues of task timing and detection sensitivity will vary depending on regional D2/D3 dopamine receptor density. Measurements of extrastriatal dopamine neuromodulation hold great promise to further our understanding of extrastriatal dopamine involvement in normal cognition and neuropsychiatric pathology. PMID- 16469509 TI - Estimating linear cortical magnification in human primary visual cortex via dynamic programming. AB - Human primary visual cortex is organized retinotopically, with adjacent locations in cortex representing adjacent locations on the retina. The spatial sampling in cortex is highly nonuniform: the amount of cortex devoted to a unit area of retina decreases with increasing retinal eccentricity. This sampling property can be quantified by the linear cortical magnification factor, which is expressed in terms of millimeters of cortex per degree of visual angle. In this paper, we present a new method using dynamic programming and fMRI retinotopic eccentricity mapping to estimate the linear cortical magnification factor in human primary visual cortex (V1). We localized cortical activity while subjects viewed each of seven stationary contrast- reversing radial checkerboard rings of equal thickness that tiled the visual field from 1.62 to 12.96 degrees of eccentricity. Imaging data from all epochs of each ring were contrasted with data from fixation epochs on a subject-by-subject basis. The resulting t statistic maps were then superimposed on a local coordinate system constructed from the gray/white matter boundary surface of each individual subject's occipital lobe, separately for each ring. Smoothed maps of functional activity on the cortical surface were constructed using orthonormal bases of the Laplace-Beltrami operator that incorporate the geometry of the cortical surface. This allowed us to stably track the ridge of maximum activation due to each ring via dynamic programming optimization over all possible paths on the cortical surface. We estimated the linear cortical magnification factor by calculating geodesic distances between activation ridges on the cortical surface in a population of five normal subjects. The reliability of these estimates was assessed by comparing results based on data from one quadrant to those based on data from the full hemifield along with a split-half reliability analysis. PMID- 16469511 TI - Aspirin "resistance". AB - A variable responsiveness to antiplatelet drugs is a clinical phenomenon that does not principally differ from other drug treatments in other therapeutic fields. The pharmacological part is to clarify whether a "true" resistance exists in pharmacological terms, i.e., a reduced potency of the compound to work as suggested and to find out the underlying cellular mechanism(s). Two principally different methods of laboratory control for platelet sensitivity to aspirin (ASA) are available: measurement of platelet function (ex vivo) or measurement of inhibition of thromboxane formation. Both methods have limitations and did not yet result in a generally accepted definition of a pharmacological ASA "resistance". The new typological approach of Weber et al. [A.A. Weber, B. Przytulski, A. Schanz, et al., Towards a definition of aspirin resistance: a typological approach. Platelets 13 (2002) 37.] helps to identify different subtypes of ASA resistance in pharmacological terms by combining in vitro aggregometry with thromboxane measurement. Using this method, a "true" pharmacological resistance, associated with a reduced antiplatelet response to ASA and reduced inhibition of thromboxane formation, was found in patients undergoing coronary artery bypass surgery. Platelets of these patients expressed a hitherto unknown isoform of COX-2-COX-2a which might generate a different gene product. In this context, it is interesting that CABG patients express transiently an immunoreactive COX-2 protein with lower molecular weight. Studies on the significance of this finding for ASA resistance are in progress. PMID- 16469512 TI - Platelet regulation by NO/cGMP signaling and NAD(P)H oxidase-generated ROS. AB - Platelets play a crucial role in the physiology of primary hemostasis and pathophysiological processes such as arterial thrombosis. Accumulating evidence suggests a key regulatory role of both NO and reactive oxygen species (ROS) in platelets. While the inhibitory role of NO/cGMP signaling in both murine and human platelets is well established, recent data suggest that intracellular ROS generation is involved in platelet activation. Thrombin-induced intracellular ROS production was inhibited by NAD(P)H oxidase inhibitors (DPI and apocynin), cyclooxygenase inhibitor (acetylsalicylic acid), and superoxide scavengers (tiron and MnTMPyP). Furthermore, thrombin (Trap6)-induced platelet aggregation and thrombus formation on collagen under high shear was inhibited by NAD(P)H oxidase inhibitors (DPI and apocynin), whereas secretion and platelet shape change were not affected. Inhibition of alphaIIbbeta3 activation by NAD(P)H oxidase inhibitors and superoxide scavengers was independent of NO/cGMP signaling demonstrating a direct role of platelet NAD(P)H oxidase-generated ROS for integrin alphaIIbbeta3 activation. PMID- 16469513 TI - Platelet interactions with vessel wall components during thrombogenesis. AB - We discuss the interrelationships of platelet glycoprotein receptors, von Willebrand factor, collagen, non-collagen matrix protein, and vascular cells in the complex events of platelet plug and subsequent thrombus formation. PMID- 16469514 TI - Selective detection of ordered sodium signals via the central transition. AB - Given the correlation between the concentrations of ordered (23)Na and the onset of tissue disorders, the ability to select the signal from ordered (23)Na over that of free (23)Na is of particular importance and can greatly enhance the potential of (23)Na-MRI as a diagnostic tool. Here, we describe a simple method that selectively detects the central transition of ordered sodium while minimizing the signal from free sodium. Our method relies upon the influence of the quadrupolar interaction on nutation frequencies and may also benefit solid state imaging experiments. Both a liquid crystalline environment and a cartilage sample are used to demonstrate a clean separation between anisotropic and isotropic regions in the experiments. PMID- 16469515 TI - LTCI, a novel chymotrypsin inhibitor of the potato I family from the earthworm Lumbricus terrestris. Purification, cDNA cloning, and expression. AB - A novel chymotrypsin inhibitor of the potato I protease inhibitor family from the earthworm Lumbricus terrestris was purified. The inhibitor, named LTCI, was isolated by methanol extraction, affinity chromatography on immobilized methylchymotrypsin, and ion exchange chromatography followed by RP-HPLC. The 7076 Da inhibitor consists of a single polypeptide chain of 64-amino-acid residues without disulfide bridges. LTCI is the first of the potato I protease inhibitors with Tyr in position P1 of the reactive site. cDNA analysis revealed that LTCI is produced as a 86-amino-acid precursor with a 22-amino-acid secretory signal peptide. RT-PCR analysis demonstrates that LTCI mRNA is expressed in body wall, intestine, and coelomocytes. The recombinant LTCI was produced in Escherichia coli as a fusion protein with intein and chitin binding domain using IMPACT-CN system. PMID- 16469516 TI - A cutaneous cyst caused by brucellosis with a negative serological test. PMID- 16469517 TI - Clinical and microbiological features of dientamoebiasis in patients suspected of suffering from a parasitic gastrointestinal illness: a comparison of Dientamoeba fragilis and Giardia lamblia infections. AB - OBJECTIVES: To describe the clinical and microbiological features of Dientamoeba fragilis and Giardia lamblia infected patients, and to analyze the genetic variation of D. fragilis strains. METHODS: For a period of two years, all stool samples collected from patients suspected of having a parasitic gastrointestinal infection were examined according to our specific triple feces test (TFT) protocol. A retrospective case-control study was performed on D. fragilis and G. lamblia infected patients. Furthermore, PCR and genotyping by restriction fragment length polymorphism (RFLP) were performed upon the former. RESULTS: D. fragilis (6.3%) and G. lamblia (7.1%) were the most common pathogenic protozoa isolated out of 448 patients studied. Symptoms most frequently encountered with D. fragilis and G. lamblia infection were abdominal pain (69.2% and 72.4%, respectively) and diarrhea (61.5% and 79.3%, respectively). However, patients with D. fragilis infections suffered significantly less frequently from nausea and/or vomiting, anorexia and weight loss. After treatment, all D. fragilis and G. lamblia infected patients presenting a negative TFT follow-up also reported a complete resolution of their symptoms. Only genotype 1 could be detected in D. fragilis infected patients. CONCLUSIONS: D. fragilis and G. lamblia were the most frequently encountered parasites in our study population. Improved diagnostic tests are essential tools to study the prevalence and pathogenesis of D. fragilis. PMID- 16469518 TI - Familial brucellosis: A report of four patients. PMID- 16469519 TI - Epidemiological pattern of classical Borna disease and regional genetic clustering of Borna disease viruses point towards the existence of to-date unknown endemic reservoir host populations. AB - Classical Borna disease (cBD), a non-purulent encephalitis of solipeds and sheep, is endemic in certain areas of central Europe. The etiologic agent is Borna disease virus (BDV), thus far the only member of the family Bornaviridae. Based on epidemiological patterns of cBD and recent phylogenetic findings this review hypothesizes the possible existence of yet unknown BDV reservoir host populations, and analyzes critically BDVs from outside endemic regions. PMID- 16469520 TI - Memory T cells protect against Plasmodium vivax infection. AB - Immunity induced by Plasmodium vivax infection leads to memory T cell recruitment activated during "relapse" or "re-infection". This study aims to characterise memory T cells in patients with acute or convalescent P. vivax infection. Lymphocytes were collected from patients infected by P. vivax, immune controls and naive controls. The proportion of immature memory T cells, expressing CD45RO(+)CD27(+), and mature cells lacking CD27 was assessed. A statistically significant increase in the median percentage of memory T cell subsets expressing CD4(+) was observed in material from patients with an acute infection compared with that from either naive or immune controls. The high percentage of memory T cells in infected patients was maintained until 60 days post treatment. The immune controls living in a malaria endemic area had a somewhat increased proportion of memory T cell subsets expressing CD8(+). An approximately three fold increase of these cell types was shown in patients with an acute infection and the level persisted until 60 days post treatment. Phenotypic characterisation of the peripheral lymphocytes during acute infection revealed that a large fraction of the lymphocytes carried the gammadelta phenotypes suggesting a role for these cells in the early response against P. vivax. Very low levels of P. vivax specific antibody were found. This might suggest that cell-mediated immunity may play a greater role in the development of naturally acquired protection against P. vivax infection than humoral immunity. Our results provide further insight into the mechanism of cell-mediated immunity to P. vivax infection that could be important for the future development of a successful vaccine and anti-malarial drug designation. PMID- 16469521 TI - Upregulation of surface proteins in Leishmania donovani isolated from patients of post kala-azar dermal leishmaniasis. AB - Five to fifteen percent of visceral leishmaniasis (VL) patients in India develop post kala-azar dermal leishmaniasis (PKDL), usually 1-2 years after apparent clinical cure. There is evidence pointing to a role played by the host immune responses in the disease pathogenesis, however, the contribution of changes in parasite gene expression has not been explored. Highly sensitive gene expression microarray technology was employed to identify genes that are differentially expressed in Leishmania parasites isolated from PKDL patients in comparison with those from VL. Hybridization on Leishmania donovani genomic microarray comprised of unique clones allowed us to identify 46/2268 (2%) clones that showed statistically significant (P<0.05) changes in expression (1.5-3.5-fold) in parasites of PKDL origin compared to those of VL origin. Sequence analysis of six genomic clones, consistently showing approximately 2-fold higher expression in PKDL parasites, revealed significant homology with gp63, gp46, putative amastin, a putative reductase and a possible calpain-like protein. The gene products showing upregulated expression in PKDL isolates may be candidates playing a role in the altered clinical manifestation in PKDL. Such differentially expressed genes hold the key to understanding the parasite genetic factors that contribute to the persistence after clinical cure of VL. PMID- 16469522 TI - Toward xeno-free culture of human embryonic stem cells. AB - The culture of human embryonic stem cells (hESCs) is limited, both technically and with respect to clinical potential, by the use of mouse embryonic fibroblasts (MEFs) as a feeder layer. The concern over xenogeneic contaminants from the mouse feeder cells may restrict transplantation to humans and the variability in MEFs from batch-to-batch and laboratory-to-laboratory may contribute to some of the variability in experimental results. Finally, use of any feeder layer increases the work load and subsequently limits the large-scale culture of human ES cells. Thus, the development of feeder-free cultures will allow more reproducible culture conditions, facilitate scale-up and potentiate the clinical use of cells differentiated from hESC cultures. In this review, we describe various methods tested to culture cells in the absence of MEF feeder layers and other advances in eliminating xenogeneic products from the culture system. PMID- 16469523 TI - Visual and oculomotor selection: links, causes and implications for spatial attention. AB - Natural scenes contain far more information than can be processed simultaneously. Thus, our visually guided behavior depends crucially on the capacity to attend to relevant stimuli. Past studies have provided compelling evidence of functional overlap of the neural mechanisms that control spatial attention and saccadic eye movements. Recent neurophysiological work demonstrates that the neural circuits involved in the preparation of saccades also play a causal role in directing covert spatial attention. At the same time, other studies have identified separable neural populations that contribute uniquely to visual and oculomotor selection. Taken together, all of the recent work suggests how visual and oculomotor signals are integrated to simultaneously select the visual attributes of targets and the saccades needed to fixate them. PMID- 16469524 TI - Neuroeconomics: cross-currents in research on decision-making. AB - Despite substantial advances, the question of how we make decisions and judgments continues to pose important challenges for scientific research. Historically, different disciplines have approached this problem using different techniques and assumptions, with few unifying efforts made. However, the field of neuroeconomics has recently emerged as an inter-disciplinary effort to bridge this gap. Research in neuroscience and psychology has begun to investigate neural bases of decision predictability and value, central parameters in the economic theory of expected utility. Economics, in turn, is being increasingly influenced by a multiple systems approach to decision-making, a perspective strongly rooted in psychology and neuroscience. The integration of these disparate theoretical approaches and methodologies offers exciting potential for the construction of more accurate models of decision-making. PMID- 16469525 TI - Perirhinal cortical contributions to object perception. AB - The traditional theory of the medial temporal lobe (MTL) memory system asserts that the primate MTL (hippocampus, perirhinal, entorhinal and parahippocampal cortices) is exclusively involved in consolidating declarative memories. However, several recent reports have directly challenged this dogma by arguing that MTL structures also contribute to perception. Controversy remains as many of the behavioural tasks used have confounded memory with perception. We review the evidence here and highlight new studies in humans and macaques that indicate a perceptual role for MTL in the absence of such confounds. We argue that the challenge to MTL memory system theory is substantiated and that the implications are considerable, namely that most psychologists and neuroscientists have held a fundamentally flawed view of how memory is implemented in the brain. PMID- 16469526 TI - Incidence of oral and oropharyngeal cancer in United Kingdom (1990-1999) -- recent trends and regional variation. AB - This study aimed to determine whether the incidence of oral cancer is continuing to rise in the UK and if this varies geographically. A descriptive epidemiological study of oral cancer incidence in 12 UK cancer registries (1990 1999) was undertaken. Poisson regression models were employed to assess trends. There were 32,852 oral cancer cases registered (1990-1999). Statistically significant increases in incidence of 18% and 30% were seen in males and females respectively (p<0.01). The trend was observed in younger (<45 years) and older (45+ years) age groups (p<0.01) with 3.5% and 2.4% average annual increases respectively. These increases were consistent for the majority of regions in the older group. For the younger group the increases in incidence were more rapid and differed geographically. Incidence remains higher in men than women, in older compared with younger groups, and in northern regions. These findings provide evidence of a continuing increase in the burden of oral cancer across the UK. PMID- 16469528 TI - Clinical features of cystadenolymphoma (Warthin's tumor) of the parotid gland: a retrospective comparative study of 96 cases. AB - The details of the etiopathogenesis of cystadenolymphoma (Warthin's tumor) are still unclear. To explore the possible risk factors for the development of this tumor, medical records of 81 patients with 96 Warthin's tumors of the parotid glands were compared retrospectively with those of 91 patients with pleomorphic adenoma. The medical history and clinical tumor characteristics of all patients were similar. There were no significant differences between these two patient groups with respect to concomitant diseases, regular medications, and preoperative laboratory findings. However, a significant male predominance of patients with Warthin's tumor could be noted (P<0.05). The male to female ratio was 3.3:1 in patients with Warthin's tumor. Multifocal Warthin's tumor were detected in five cases (6.2%), and 10 patients (12.3%) had bilateral lesions. The odds ratio for the incidence of Warthin's tumor among current smokers compared with never smokers was 8.3 (P<0.0001). Compared with never smokers, clearly higher odds of Warthin's tumor was observed in heavy smokers (more than 30 pack years) (odds ratio=24.1, P<0.0001) than patients who smoked less than 30 pack years (odds ratio=4.9, P<0.0001). PMID- 16469527 TI - Hypoxia enhances c-Met/HGF receptor expression and signaling by activating HIF 1alpha in human salivary gland cancer cells. AB - Hypoxia increases the invasive and metastatic potential of tumor cells. Increased expression of c-Met/hepatocyte growth factor (HGF)-receptor protein in response to hypoxia in thyroid papillary carcinomas is hypoxia-inducible factor-1 (HIF-1) dependent. Both HGF and c-Met are expressed in human salivary gland cancers. In the current study, we tested whether c-Met expression was regulated by hypoxia and HIF-1alpha using two human salivary gland cancer cell lines: GFP-ACC2 and GFP ACCM. Hypoxia enhanced the expression of HIF-1alpha in both cell lines, whereas c Met was markedly induced only in the GFP-ACCM cells, which have metastatic potential. In the latter, hypoxia also promoted HGF-induced invasiveness. Synthetic small-interfering RNA specific for HIF-1alpha inhibited HIF-1alpha expression in the GFP-ACCM cells, and also suppressed the increase in c-Met expression and HGF-induced invasiveness under hypoxic conditions. These results suggest that hypoxia activates the HGF/c-Met system via HIF-1alpha in human salivary gland cancers and might be involved in their metastasis. PMID- 16469530 TI - Orthogonal polarization spectral (OPS) imaging and topographical characteristics of oral squamous cell carcinoma. AB - Tumor microcirculatory characteristics until now have only been assessed by histological examination of biopsies or invasive imaging technique. The recent introduction of orthogonal polarization spectral (OPS) imaging as a new tool for in vivo visualization of human microcirculation makes it possible to acquire high resolution images of the oral mucosa. In this study we report the microcirculatory changes in ten patients with oral squamous cell carcinoma of the tongue and compared the images to the normal contralateral side as the control. All carcinomas were T2 tumors without evidence for lymph node metastasis. The carcinomas were characterized by chaotic and dilated vessels accompanied by numerous areas of haemorrhage. The OPS technique seems very promising in the assessment of oral squamous cell carcinoma microcirculatory characteristics and may possibly play a future role in both the detection of early oral mucosal vascular aberrations and the effect of anti-tumor agents on the tumor microvasculature. PMID- 16469529 TI - Eosinophilic granuloma of the jaws: an analysis of clinical and radiographic presentation. AB - Eosinophilic granuloma (EG) is the localized and mildest form of histiocytosis-X group of diseases. In this study, 22 patients of pathologically proven EG involving the jaws were retrospectively studied concerning the clinical and radiographic presentation. The clinical and radiographic presentation of the 22 cases was quite varied, and the accuracy of clinical and radiographic diagnosis was low. However, the clinical and radiographic presentation correlated with the lesion site, and accordingly all the jaw lesions could be divided into three types (Alveolar Type, Intraosseous Type, and Mixed Type). Our classification based on the lesion sites reveals the correlation between the clinical and radiographic presentation of EG and the lesion sites, and may be useful in the identification of lesions of EG in the jaws. PMID- 16469531 TI - Predictors of urinary tract infection based on artificial neural networks and genetic algorithms. AB - BACKGROUND: Among women who present with urinary complaints, only 50% are found to have urinary tract infection. Individual urinary symptoms and urinalysis are not sufficiently accurate to discriminate those with and without the diagnosis. METHODS: We used artificial neural networks (ANN) coupled with genetic algorithms to evolve combinations of clinical variables optimized for predicting urinary tract infection. The ANN were applied to 212 women ages 19-84 who presented to an ambulatory clinic with urinary complaints. Urinary tract infection was defined in separate models as uropathogen counts of > or =10(5) colony-forming units (CFU) per milliliter, and counts of > or =10(2) CFU per milliliter. RESULTS: Five variable sets were evolved that classified cases of urinary tract infection and non-infection with receiver-operating characteristic (ROC) curve areas that ranged from 0.853 (for uropathogen counts of > or =10(5) CFU per milliliter) to 0.792 (for uropathogen counts of > or =10(2) CFU per milliliter). Predictor variables (which included urinary frequency, dysuria, foul urine odor, symptom duration, history of diabetes, leukocyte esterase on urine dipstick, and red blood cells, epithelial cells, and bacteria on urinalysis) differed depending on the pathogen count that defined urinary tract infection. Network influence analyses showed that some variables predicted urine infection in unexpected ways, and interacted with other variables in making predictions. CONCLUSIONS: ANN and genetic algorithms can reveal parsimonious variable sets accurate for predicting urinary tract infection, and novel relationships between symptoms, urinalysis findings, and infection. PMID- 16469532 TI - Alteration of serum cytokine balances among different phases of chronic hepatitis B virus infection. AB - To understand the pathogenesis of chronic hepatitis B virus (HBV) infection, we examined the serum levels of IL-10, TNF-alpha IL-12 p70, and IL-12 p40 in 77 patients chronically infected with HBV and 19 controls. The patients were classified into four groups: asymptomatic carriers (ASC), patients with chronic hepatitis (CH), patients with liver cirrhosis (LC), and patients with hepatocellular carcinomas (HCC). The cytokine values among these groups were compared and their relations to clinical parameters were investigated. All these cytokine values were higher in the patient groups than in controls. IL-10 and TNF alpha became higher in accordance with the progress of the disease phases, from ASC to LC, and lowest when the patients had HCC. IL-12 p40 was also lowest in HCC, however, the group with highest levels was CH. IL-12 p70 was unchanged among ASC, CH, and LC, but were raised in HCC. Serial analyses for the cytokine values in the same patients showed the similar tendencies. Regression analysis showed the significant correlations between ALT and IL-10. Serum cytokine values well reflected the pathological differences of the individual disease phases, and may become useful indices to understand the pathogenesis of chronic HBV infection. PMID- 16469533 TI - Signalling diacylglycerol pyrophosphate, a new phosphatidic acid metabolite. AB - Diacylglycerol pyrophosphate (DGPP) is a novel phospholipid that has been found in plants and yeast but not in higher animals. It is produced through phosphorylation of phosphatidic acid (PA) by the novel enzyme PA kinase (PAK). In plants, DGPP is virtually absent in non-stimulated cells but its concentration increases within minutes in response to various stimuli, including osmotic stress and pathogen attack, implying a role in stress signalling. DGPP is broken down by the enzyme DGPP phosphatase (DPP). DPP-encoding genes have been cloned from Arabidopsis thaliana and Saccharomyces cerevisiae (DPP1). In S. cerevisiae, the expression of DPP1 is regulated coordinately with the majority of genes encoding enzymes involved in phospholipid biosynthesis. PMID- 16469534 TI - Emergent EEG is helpful in neurology critical care practice. PMID- 16469536 TI - Tei index and neurohormonal activation in patients with incident heart failure: serial changes and prognostic value. AB - BACKGROUND: Natriuretic peptides and Tei index are useful indices for risk stratification in advanced left ventricular dysfunction (LVD). Their role in early stages is less clear. AIMS: In relation to first diagnosis of LVD to assess the relation of plasma B-type-natriuretic peptide (NT-proBNP) with Tei index, assess serial changes in indices, and to assess the value of indices to predict functional status. METHODS: Doppler echocardiography and neurohormonal analysis were performed (n=150). NYHA class was registered. RESULTS: Tei index correlated with p-NT-proBNP (r=0.75, p<0.0001), and changes in indices correlated (r=0.36, p=0.001) in LVD (n=80). No functional improvement (n=47) was related to a median increase in Tei index (-0.2, -0.16; 0.09); an improvement (n=31) to a reduction (0.06; -0.19; 0.35), p=0.02. In the group with functional improvement, more patients had >/=30% reduction in p-NT-proBNP (75% vs. 45%, p<0.01). Addition of NT-proBNP or Tei index to a clinical model, of no functional improvement, improved log-likelihood chi(2) from 9.32 to 20.18 (p=0.001) and 20.67 (p=0.001). CONCLUSION: Tei index and p-NT-proBNP demonstrated a fair correlation. Unimproved NYHA class was related to progressive LVD and might be identified by monitoring Tei index or p-NT-proBNP. Advanced LVD and high pre-treatment p-NT-proBNP levels indicated a potential of improvement in functional status. PMID- 16469538 TI - Effects of autoantibodies removed by immunoadsorption from patients with dilated cardiomyopathy on neonatal rat cardiomyocytes. AB - INTRODUCTION: Immunoadsorption has been shown to improve cardiac performance and reduce mortality in patients with dilated cardiomyopathy. In this study, the underlying mechanism for these beneficial effects was investigated in cultured rat cardiomyocytes. METHODS AND RESULTS: Immunoadsorption was performed in patients with dilated cardiomyopathy (n=7). Antibody-induced complement-dependent cytotoxicity was investigated by colorimetric MTT. Autoantibodies against the beta(1)-adrenoceptor were detected by ELISA and purified. Column eluent from six patients exhibited a cytotoxic effect, three patients were positive for the beta(1)-adrenoceptor autoantibodies. The purified autoantibodies were able to visualize the beta(1)-adrenoceptors by immunocytofluorescence on rat cardiomyocytes, and also displayed partial agonist properties and induced a positive chronotropic effect, which were blocked by the beta(1)-selective antagonist bisoprolol and the peptide corresponding to the beta(1)-adrenoceptor. Column eluent from one patient induced apoptosis in nick end labelling test (8.1+/-1.7% vs. 2.9+/-1.2% in control, p<0.05). CONCLUSION: Autoantibodies removed by immunoadsorption from patients with dilated cardiomyopathy have a pathophysiological role, as shown by the complement-dependent cytotoxicity and chronotropic action on rat cardiomyocytes. This implies that removal of circulating autoantibodies might be part of the underlying mechanism for improved cardiac function. PMID- 16469539 TI - Fructose 1,6-diphosphate administration attenuates post-ischemic ventricular dysfunction. AB - BACKGROUND: Cardiomyocyte energy production during ischemia depends upon anaerobic glycolysis inefficiently yielding two ATP per glucose. Substrate augmentation with fructose 1,6-diphosphate (FDP) bypasses the ATP consuming steps of glucokinase and phosphofructokinase thus yielding four ATP per FDP. This study evaluated the impact of FDP administration on myocardial function after acute ischemia. METHODS: Male Wistar rats, 250-300 g, underwent 30 min occlusion of the left anterior descending coronary artery followed by 30 min reperfusion. Immediately prior to both ischemia and reperfusion, animals received an intravenous bolus of FDP or saline control. After 30 min reperfusion, myocardial function was evaluated with a left ventricular intracavitary pressure/volume conductance microcatheter. For bioenergetics studies, myocardium was isolated at 5 min of ischemia and assayed for ATP levels. RESULTS: Compared to controls (n=8), FDP animals (n=8) demonstrated significantly improved maximal left ventricular pressure (100.5+/-5.4 mmHg versus 69.1+/-1.9 mmHg; p<0.0005), dP/dt (5296+/-531 mmHg/s versus 2940+/-175 mmHg/s; p<0.0028), ejection fraction (29.1+/ 1.7% versus 20.4+/-1.4%; p<0.0017), and preload adjusted maximal power (59.3+/ 5.0 mW/microL(2) versus 44.4+/-4.6 mW/microL(2); p<0.0477). Additionally, significantly enhanced ATP levels were observed in FDP animals (n=5) compared to controls (n=5) (535+/-156 nmol/g ischemic tissue versus 160+/-9.0 nmol/g ischemic tissue; p<0.0369). CONCLUSIONS: The administration of the glycolytic intermediate, FDP, by intravenous injection, resulted in significantly improved myocardial function after ischemia and improved bioenergetics during ischemia. PMID- 16469540 TI - Loss of autoimmune T cells correlates with brain diseases: possible implications for schizophrenia? AB - T-cell-mediated autoimmunity participates in physiological defense, maintenance and repair of the adult brain. However, unless such autoimmune responses to insults are rigorously controlled, they might lead to an autoimmune disease or other immune-related defects, where destructive activity outweighs the beneficial effect. Here, we discuss these apparently contradictory effects of autoimmunity in schizophrenic patients, whose typical immune aberrations have prompted recent speculation about an autoimmune-related etiology. We found that, although schizophrenic patients have active immune systems, they often lack autoimmune clones specifically reactive to a major myelin protein, myelin basic protein (MBP). This, in conjunction with our discovery in rodents that T cells that recognize brain-resident proteins are needed for normal cognitive functioning, led us to propose an immune-based neurodevelopmental hypothesis, in which autoimmune-T-cell deficiency is suggested to cause onset or progression of schizophrenia. PMID- 16469542 TI - Human rights aspects of safe motherhood. AB - Maternal deaths in developing countries are often the ultimate tragic outcome of the cumulative denial of women's human rights. Women are not dying because of untreatable diseases. They are dying because societies have yet to make the decision that their lives are worth saving. Maternity is a social function and not a disease. When women are risking death to give life, they are entitled to have their own right to life and health protected. Societal attitudes of looking at women as means and not ends have resulted in the denial of women's rights to essential maternity services. A signal of hope is that safe motherhood is now on the world agenda as one of eight Millennium Development Goals. The global community of obstetricians has a major responsibility to help make motherhood safer for all women. PMID- 16469541 TI - Metabolic therapeutics in angina pectoris: history revisited with perhexiline. AB - The ever-increasing burden of ischaemic heart disease and its common manifestation chronic angina pectoris calls for the exploration of other treatment options for those patients who despite the maximum conventional pharmacological and surgical interventions continue to suffer. Such exploration has led to the increasing use of new metabolically acting antianginal agents and the re-emergence of an old and somewhat forgotten pharmacological agent, perhexiline maleate. This review aims to update the cardiac nurse with knowledge to manage the care a patient receiving perhexiline maleate treatment and provide a brief review of three new metabolic agents: trimetazidine, ranolazine and etomoxir. PMID- 16469543 TI - Selective intratumoral amplification of an antiangiogenic vector by an oncolytic virus produces enhanced antivascular and anti-tumor efficacy. AB - The development of effective cancer therapy will require the simultaneous targeting of multiple steps in tumor development. We have previously described an antiangiogenic gene therapy vector, Ad Flk1-Fc, which expresses a soluble VEGF receptor capable of inhibiting tumor angiogenesis and growth. We have also described an oncolytic virus, dl922/947, whose replication and subsequent cytotoxicity are restricted to cancer cells with a loss of the G1-S cell cycle checkpoint. Here we have optimized methods for combining these therapies, yielding significantly greater anti-tumor effects than the respective monotherapies. In cultured tumor lines, co-infection with both Ad Flk1-Fc and dl922/947 allowed replication and repackaging of the replication-deficient Ad Flk1-Fc and enhanced soluble VEGF receptor expression. Similar repackaging and increased gene expression were demonstrated in vivo using bioluminescence imaging studies. Finally, coadministration of these therapeutic viral therapies in vivo produced significantly enhanced anti-tumor effects in colon HCT 116 and prostate PC-3 xenografts in mice. This increased therapeutic benefit correlated with replication of Ad Flk1-Fc viral genomes, increased intratumoral levels of Flk1-Fc protein, and decreased microvessel density, consistent with enhanced antiangiogenic activity. PMID- 16469544 TI - The Na+, K+, 2Cl- cotransporter of estuarine pufferfishes (Sphoeroides testudineus and S. greeleyi) in hypo- and hyper-regulation of plasma osmolality. AB - The pufferfishes Sphoeroides testudineus and Sphoeroides greeleyi are estuarine species that osmoregulate efficiently, but S. testudineus tolerates seawater dilution to a much higher degree than S. greeleyi. This study aimed at testing whether NKCC is involved with their differential tolerance of seawater dilution, through the analysis of in vivo furosemide (NKCC inhibitor) injection both on hypo-regulation (in 35 per thousand salinity) and hyper-regulation (in 5 per thousand salinity). After exposure for 6 h or 5 days to both salinities, blood samples were obtained for determination of plasma osmolality, chloride, sodium and hematocrit, and muscle samples for determination of water content. Furosemide injection led to increased plasma osmolality and sodium in 35 per thousand and decreased osmolality and chloride in 5 per thousand, when compared to saline injected controls. Furosemide injection led to hematocrit reduction in both salinities, and muscle water content increase in 5 per thousand and decrease in 35 per thousand in S. testudineus. The results are compatible with NKCC working in branchial NaCl secretion in 35 per thousand, in both species, and a higher role in cell volume regulation in blood and muscle cells of S. testudineus, in both salinities, which could partially explain the stronger capacity of S. testudineus to tolerate seawater dilution during low tide. PMID- 16469545 TI - Using hit curves to compare search algorithm performance. AB - Databases continue to grow but the metrics available to evaluate information retrieval systems have not changed. Large collections such as MEDLINE and the World Wide Web contain many relevant documents for common queries. Ranking is therefore increasingly important and successful information retrieval systems, such as Google, have emphasized ranking. However, existing evaluation metrics such as precision and recall, do not directly account for ranking. This paper describes a novel way of measuring information retrieval performance using weighted hit curves adapted from the field of statistical detection to reflect multiple desirable characteristics such as relevance, importance, and methodologic quality. In statistical detection, hit curves have been proposed to represent occurrence of interesting events during a detection process. Similarly, hit curves can be used to study the position of relevant documents within large result sets. We describe hit curves in light of a formal model of information retrieval, show how hit curves represent system performance including ranking, and define ways to statistically compare performance of multiple systems using hit curves. We provide example scenarios where traditional measures are less suitable than hit curves and conclude that hit curves may be useful for evaluating retrieval from large collections where ranking performance is crucial. PMID- 16469546 TI - Cardiovascular autonomic nervous response to postural change in 610 healthy Japanese subjects in relation to age. AB - To determine the effect of aging on the cardiovascular response to postural change, we examined the cardiovascular sympathetic and parasympathetic response to active standing in 610 healthy Japanese subject (6-83 years) measuring the initial heart rate (HR) response for 3 min in the supine and standing position, we also measured the coefficient of variation of R-R interval (CV(R-R)). As a result, the cardiovascular response to active standing demonstrated a different change with aging between sympathetic and parasympathetic. Sympathetic function was in a sthenia state in young subjects, and that this function declined with age increasing. Whereas, parasympathetic function was immature enough to inhibit the sympathetic tone in young subjects and matured at 20 years of age, and had an ability to inhibit sympathetic tone. CV(R-R) show a linear change that decline with age increasing. These results indicated that the cardiovascular parasympathetic response to active standing shows a characteristic change with aging that differs from cardiovascular parasympathetic at rest represented by CV(R-R). The present study is the first report to demonstrate the cardiovascular response to standing in relation to aging in large population. These results suggested that the cardiovascular response to postural change is dependent on subject's age. PMID- 16469547 TI - Purification of capsular polysaccharide from Neisseria meningitidis serogroup C by liquid chromatography. AB - Neisseria meningitidis serogroup C capsular polysaccharide (MenCPS) is an important antigen against meningococcal infection. This paper describes a new purification methodology employing liquid chromatography that resulted in a polysaccharide showing the characteristics recommended by the World Health Organization for vaccine purposes. In this method, steps of the traditional procedure that yield low recovery and use toxic materials were modified. The present process consists in the following steps: (1) continuous flow centrifugation of the culture for removal of the cells; (2) supernatant concentration by tangential filtration (100 kDa cutoff); (3) addition of 0.5% DOC, heating to 55 degrees C during 30 min and tangential filtration (100 kDa cutoff); (4) anion exchange chromatography (Source 15Q) and (5) size exclusion chromatography (Sepharose CL-4B). The polysaccharide C fraction obtained in that way was dialyzed and freeze-dried. The structural identity of the polysaccharide was demonstrated by (1)H-NMR spectrometry. PMID- 16469548 TI - A molecular dynamics simulation of SNase and its hydration shell at high temperature and high pressure. AB - Temperature- and pressure-induced unfolding of staphylococcal nuclease (SNase) was studied by Royer, Winter et al. using a variety of experimental techniques (SAXS, FT-IR and fluorescence spectroscopy, DSC, PPC, densimetry). For a more detailed understanding of the underlying mechanistic processes of the different unfolding scenarios, we have carried out a series of molecular dynamics (MD) computer simulations on SNase. We investigated the initial changes of the structure of the protein upon application of pressure (up to 5 kbar) and discuss volumetric and structural differences between the native and pressure pre denatured state. Additionally, we have obtained the compressibility of the protein and hydration water and compare these data with experimental results. As water plays a crucial role in determining the structure, dynamics and function of proteins, we undertook a detailed analysis of the structure of the interfacial water and the protein-solvent H-bond network as well. Moreover, we report here also MD results on the temperature-induced unfolding of SNase. The time evolution of the protein volume and solvent accessible surface area during thermal unfolding have been investigated, and we present a detailed discussion of the temperature-induced unfolding pathway of SNase in terms of secondary and tertiary structural changes. PMID- 16469549 TI - Genistein alone and in combination with the mammalian lignans enterolactone and enterodiol induce estrogenic effects on bone and uterus in a postmenopausal breast cancer mouse model. AB - The use of phytoestrogens, such as isoflavones and lignans, for treatment of postmenopausal breast cancer is increasing, but their effects on bone and other major organs are not clear. While the isoflavone genistein (GEN) has been shown to prevent or slow the loss of bone mineral density (BMD), the effect of lignans enterodiol (END) and enterolactone (ENL) are unknown. In this study, we determined in ovariectomized mice with human MCF-7 breast tumor xenografts the effects of the lignans, and GEN, alone and in combination, on bone and uterus. Mice with established MCF-7 tumors were fed a basal diet (AIN-93G), divided into 5 groups, and given daily subcutaneous injections (10 mg/kg body weight) of either ENL, END, GEN, a mixture of these compounds (MIX), or vehicle as a negative control for 22 weeks. Results showed that GEN acts estrogenically in both the uterus and bone by increasing the uterus weight, femur BMD, and femur biomechanical strength (yield load), while the lignans do not. However, treatment with MIX induced minimal effects on femur biomechanical strength parameters but significantly increased uterus weight. A significant positive correlation was observed between MCF-7 tumor volume and femur BMD and biomechanical strength parameters (femur peak load and yield load) but not with uterus weight, suggesting that the uterus may respond differently to phytoestrogens compared to MCF-7 tumors and bone. It is concluded that GEN induces beneficial effects on bone but has adverse effects on tumors and uterus in this model of postmenopausal breast cancer. The lignans do not exert adverse effects on any tissue, however, when combined with GEN, they exert an adverse effect on the uterus. PMID- 16469550 TI - Ornithine alpha-ketoglutarate increases mineralization and mechanical properties of tibia in turkeys. AB - Skeletal disorders in rapidly growing poultry are commonplace. This study was performed to investigate the effect of ornithine alpha-ketoglutarate (OKG) administration during the last 7 weeks of life on structural properties, mineralization, and mechanical endurance of skeleton in turkeys at slaughter. Healthy HB Medium Bronze female turkeys were randomly assigned to two weight matched groups at the age of 12 weeks. OKG was administered orally to the experimental group (N=17) at the dose of 0.4 g/kg body weight per day, while the control group (N=16) received an equal dose of the vehicle. The turkeys were slaughtered at the age of 19 weeks and the tibiae were isolated for analysis. The effect of OKG on skeletal system development in turkeys was evaluated in relation to both geometrical and mechanical properties as well as quantitative computed tomography (QCT). Free amino acids concentrations were assessed with the use of ion-exchange chromatography. Significantly increased bone mineral density of the trabecular and the cortical bone of tibia in the turkeys given OKG for the last 7 weeks of production cycle were observed (P<0.05). OKG administration improved mechanical endurance of the tibia estimated by the three-point bending test (P<0.01). Plasma amino acid analyses showed increased level of aspartate, proline, alanine, valine, isoleucine, leucine, and ornithine (all P<0.05) after OKG treatment, whereas cystathionine concentration was decreased (P=0.03). Obtained results indicate that oral OKG administration has beneficial effects on skeletal development in fast growing turkeys and this effect is connected with increased amino acid synthesis. These observations may serve to improve skeletal properties in birds, especially when considering that skeletal disorders often affect the tibia and the proper function of the skeletal system plays an essential role in animal welfare and poultry production. PMID- 16469555 TI - Modifying urology residency training: time to speed up or slow down? PMID- 16469551 TI - Shear stress facilitates tissue-engineered odontogenesis. AB - Numerous studies have demonstrated the effect of shear stress on osteoblasts, but its effect on odontogenic cells has never been reported. In this study, we focused on the effect of shear stress on facilitating tissue-engineered odontogenesis by dissociated single cells. Cells were harvested from the porcine third molar tooth at the early stage of crown formation, and the isolated heterogeneous cells were seeded on a biodegradable polyglycolic acid fiber mesh. Then, cell-polymer constructs with and without exposure to shear stress were evaluated by in vitro and in vivo studies. In in vitro studies, the expression of both epithelial and mesenchymal odontogenic-related mRNAs was significantly enhanced by shear stress for 2 h. At 12 h after exposure to shear stress, the expression of amelogenin, bone sialoprotein and vimentin protein was significantly enhanced compared with that of control. Moreover, after 7 days, alkaline phosphatase activity exhibited a significant increase without any significant effect on cell proliferation in vitro. In vivo, enamel and dentin tissues formed after 15 weeks of in vivo implantation in constructs exposure to in vitro shear stress for 12 h. Such was not the case in controls. We concluded that shear stress facilitates odontogenic cell differentiation in vitro as well as the process of tooth tissue engineering in vivo. PMID- 16469556 TI - Prognostic factors for renal cell carcinoma: integrating laboratory and molecular factors. PMID- 16469557 TI - Insights into the natural history and treatment of benign prostatic hyperplasia. PMID- 16469558 TI - Botulinum toxin and cellular proliferation in the prostate. PMID- 16469559 TI - Varicoceles. PMID- 16469560 TI - Prostate needle biopsies containing prostatic intraepithelial neoplasia or atypical foci suspicious for carcinoma: implications for patient care. AB - PURPOSE: We identified information critical for patient treatment on prostate needle biopsies diagnosed with prostatic intraepithelial neoplasia or atypical foci suspicious for carcinoma. MATERIALS AND METHODS: A search was performed using the MEDLINE database and referenced lists of relevant studies to obtain articles addressing the significance of finding PIN or atypical foci suspicious for carcinoma on needle biopsy. RESULTS: There were certain results concerning PIN. 1) Low grade PIN should not be documented in pathology reports due to poor interobserver reproducibility and a relatively low risk of cancer following re biopsy. 2) The expected incidence of HGPIN on needle biopsy is between 5% and 8%. 3) Although the diagnosis of HGPIN is subjective, interobserver reproducibility for its diagnosis is fairly high among urological pathologists, and yet only moderate among pathologists without special expertise in prostate pathology. 4) The median risk recorded in the literature for cancer following the diagnosis of HGPIN on needle biopsy is 24.1%, which is not much higher than the risk reported in the literature for repeat biopsy following a benign diagnosis. 5) The majority of publications that compared the risk of cancer in the same study following a needle biopsy diagnosis of HGPIN to the risk of cancer following a benign diagnosis on needle biopsy show no differences between the 2 groups. 6) Clinical and pathological parameters do not help stratify which men with HGPIN are at increased risk for a cancer diagnosis. 7) A major factor contributing to the decreased incidence of cancer following a diagnosis of HGPIN on needle biopsy in the contemporary era is related to increased needle biopsy core sampling, which detects many associated cancers on initial biopsy, such that re-biopsy, even with good sampling, does not detect many additional cancers. 8) It is recommended that men do not need routine repeat needle biopsy within the first year following the diagnosis of HGPIN, while further studies are needed to confirm whether routine repeat biopsies should be performed several years following a HGPIN diagnosis on needle biopsy. There were certain results concerning atypical glands suspicious for carcinoma. 1) An average of 5% of needle biopsy pathology reports are diagnosed as atypical glands suspicious for carcinoma. 2) Cases diagnosed as atypical have the highest likelihood of being changed upon expert review and urologists should consider sending such cases for consultation in an attempt to resolve the diagnosis as definitively benign or malignant before subjecting the patient to repeat biopsy. 3) Ancillary techniques using basal cell markers and AMACR (alpha-methyl-acyl-coenzyme A racemase) can decrease the number of atypical diagnoses, and yet one must use these techniques with caution since there are numerous false-positive and false-negative results. 4) The average risk of cancer following an atypical diagnosis is approximately 40%. 5) Clinical and pathological parameters do not help predict which men with an atypical diagnosis have cancer on repeat biopsy. 6) Repeat biopsy should include increased sampling of the initial atypical site, and adjacent ipsilateral and contralateral sites with routine sampling of all sextant sites. Therefore, it is critical for urologists to submit needle biopsy specimens in a manner in which the sextant location of each core can be determined. 7) All men with an atypical diagnosis need re-biopsy within 3 to 6 months. CONCLUSIONS: It is critical for urologists to distinguish between a diagnosis of HGPIN and that of atypical foci suspicious for cancer on needle biopsy. These 2 entities indicate different risks of carcinoma on re-biopsy and different recommendations for followup. PMID- 16469561 TI - Efficacy and safety of sacral nerve stimulation for urinary urge incontinence: a systematic review. AB - PURPOSE: We systematically reviewed the evidence on the efficacy and safety of sacral nerve stimulation (SNS) for severe urge incontinence. MATERIALS AND METHODS: A systematic review was performed of primary studies of SNS for urge incontinence published in English between 1966 and May 2003, and identified in major electronic databases. Two reviewers independently selected studies, assessed their methodological quality and extracted data. RESULTS: Four randomized controlled trials and 30 case series were identified. Evidence from the randomized controlled trials, involving approximately 120 patients, showed that about 80% achieved continence or greater than 50% improvement in their main incontinence symptoms after SNS compared with about 3% of controls receiving conservative treatments while waiting for an implant. While case series were larger, they were methodologically less reliable. However, they showed similar results with 67% of patients becoming dry or achieving a greater than 50% improvement in symptoms after implantation. Incontinence episodes, leakage severity, voiding frequency and pad use were significantly lower after implantation. Benefits were reported to persist 3 to 5 years after implantation. Adverse events were documented in 27 studies. Overall the reoperation rate in implanted cases was 33%. The most common reason for surgical revision was relocation of the generator because of pain and infection. Common complications were pain at the implant or lead site in 25% of patients, lead related problems such as lead migration in 16%, replacement and repositioning of the implanted pulse generator in 15%, wound problems in 7%, adverse effects on bowel function in 6%, infection in 5% and generator problems in 5%. Permanent removal of the electrodes was reported in 9% of patients. Technical changes with time have been associated with decreased complication rates. CONCLUSIONS: There is evidence indicating that SNS is effective for decreasing symptoms in patients with urge incontinence. Adverse events occurred in about half of the implanted cases and surgical revision was performed in 33%. No major irreversible complications were reported in the studies reviewed. Further research is required on the long-term effects of and quality of life in patients with SNS. PMID- 16469562 TI - Prevalence, characteristics and implications of premature ejaculation/rapid ejaculation. AB - PURPOSE: Premature ejaculation/rapid ejaculation is a common but incompletely understood male sexual dysfunction. The purposes of this review are to 1) raise awareness of the prevalence and characteristics of PE/RE, its impact on the male and his partner, and the lack of approved medications indicated for its treatment, 2) encourage dialogue about PE/RE between physicians and patients, and 3) stimulate the development of appropriate new therapies. MATERIALS AND METHODS: A MEDLINE search was performed to retrieve articles relating to PE/RE pathophysiology, etiology, impact, diagnosis and treatment. Sexual medicine journals not indexed in MEDLINE, sexual medicine texts and congress abstracts were also reviewed. RESULTS: No universally accepted definition, licensed treatment, validated screening instrument or diagnostic criteria have been established for PE/RE, and its pathophysiology and etiology are incompletely understood. Additional barriers are the reluctance of patients and physicians to talk about PE/RE and the lack of knowledge regarding available treatments. Current pharmacological treatments include off label uses of antidepressants, topical anesthetics or phosphodiesterase-5 inhibitors. All are associated with drawbacks that limit their efficacy. Psychological counseling and behavioral therapy have a valuable role, although resources for this modality are limited. CONCLUSIONS: Prevalence rates of 20% to 30% and negative effects on the quality of life of men and their partners illustrate the need for improved, standardized methods of PE/RE diagnosis, assessment and treatment. Medications indicated specifically for PE/RE and effective on an as needed basis are required. Behavioral therapies should emphasize pleasure, arousal, control, confidence and satisfaction, and they may have the best success when coupled with pharmacological approaches. PMID- 16469563 TI - Laparoscopic partial nephrectomy for centrally located renal tumors. AB - PURPOSE: LPN is frequently reserved for small, peripherally located tumors. Centrally located tumors typically require complex intracorporeal suturing and reconstruction with hilar clamping, which is a laparoscopically advanced maneuver given the constraints of renal ischemia. We retrospectively compared our experience with central vs peripheral tumors treated with LPN. MATERIALS AND METHODS: Between January 2001 and March 2004, 363 patients underwent LPN for tumor. The tumor was located centrally in 154 patients and peripherally in 209. Central tumors were defined as tumors centrally extending into the kidney in direct contact with or invading into the pelvicaliceal system and/or renal sinus on preoperative 3-dimensional computerized tomography. Lesions with no contact with the pelvicaliceal system were classified as peripheral. Preoperative, intraoperative, postoperative and pathological data were compared. RESULTS: Central tumors were larger (median 3 vs 2.4 cm, p < 0.001) and had larger specimens at surgery (median 43 vs 22 gm, p < 0.001) than peripheral tumors. Although blood loss was similar (median 150 cc), central tumors required longer warm ischemia time (median 33.5 vs 30 minutes, p < 0.001), operative time (median 3.5 vs 3 hours, p = 0.008) and hospital stay (median 67 vs 60 hours, p < 0.001). A positive cancer margin occurred in 1 patient per group. Median postoperative serum creatinine was similar (1.2 vs 1.1 mg/dl). Intraoperative and late postoperative complications were comparable. However, more early postoperative complications occurred in the central group (6% vs 2%, p = 0.05). CONCLUSIONS: LPN for central tumors can be performed safely by an experienced laparoscopic surgeon with perioperative outcomes comparable to those of peripheral tumors. Given the requisite laparoscopic expertise, indications for LPN should be expanded to include centrally located tumors. PMID- 16469564 TI - Partial nephrectomy for small renal masses: an emerging quality of care concern? AB - PURPOSE: The recent popularization of laparoscopic radical nephrectomy may beget underuse of partial nephrectomy. To evaluate this concern we used the SEER registry to characterize national practice patterns for the surgical management of small renal masses. MATERIALS AND METHODS: Between 1988 and 2001, 14,647 patients with primary tumor size 7 cm or less were treated surgically for locoregional kidney cancer. The proportion of patients treated with PN was determined and stratified by year of diagnosis and tumor size. Multivariate models were developed to identify independent determinants of PN use and overall survival following surgical treatment of kidney cancer. RESULTS: Overall 1,401 patients (9.6%) were treated with PN vs 13,246 (90.4%) who underwent total nephrectomy. For tumors 7 cm or less, the use of PN increased progressively between 1988 (4.6%) and 2001 (17.6%, p < 0.001). Despite this trend PN remained fairly uncommon even for the smallest renal masses. Among patients with tumors less than 2 cm, 14% underwent PN in 1988 to 1989 vs 42% in 2000 to 2001. For tumors 2 to 4 cm the corresponding proportions were 5% and 20%, respectively (p < 0.001). Younger patient age, smaller tumor size and more recent diagnostic year were independent determinants of PN use (all p values < 0.05). All cause mortality was similar for patients treated with PN vs TN (HR 0.9, 95% CI 0.8 1.1). CONCLUSIONS: Despite more frequent application during the last 2 decades, nationwide use of PN remains relatively uncommon, even for the smallest renal masses. Recognizing the favorable outcomes associated with preservation of renal parenchyma, our findings identify a possible quality of care concern that should be addressed by the urological community. PMID- 16469566 TI - Prognostic value of thrombocytosis in renal cell carcinoma. AB - PURPOSE: We correlated platelet count to renal tumor characteristics, and evaluated the potential prognostic value of thrombocytosis in localized and metastatic tumors. MATERIALS AND METHODS: A total of 804 patients operated on for a renal tumor in 2 French centers were included in this study. In all cases TNM stage, Fuhrman grade, tumor size, nodal invasion, ECOG score, histological subtype and platelet count were recorded. Survival estimates were compared with the Kaplan-Meier method and multivariate analysis was performed using a Cox model. RESULTS: There were 538 (66.9%) males and 266 (33%) females (ratio 2.02) in the patient group. Median age was 62 years. T stages were T1, T2, T3 and T4 in 284 (35.3%), 116 (14.4%), 387 (48.1%) and 17 (2.1%), respectively. There were 112 (13.9%) and 126 (15.7%) patients with nodal and metastatic invasion, respectively. Platelet count was strongly correlated with T stage (p < 0.001), Fuhrman grade (p < 0.001), tumor size (p < 0.001), nodal invasion (p < 0.001) and the presence of distant metastasis (p = 0.01). In univariate analysis all studied factors were significantly predictive of survival (p = 0.0001). Thrombocytosis had an impact on prognosis in localized (p < 0.001) and metastatic (p = 0.003) disease. Several factors were retained in multivariate analysis, namely TNM stage, Fuhrman grade, tumor size, ECOG score and platelet count. In cases with a platelet count of less than 450,000/mm3, the 5-year survival rate was 70%, compared to 38% when the platelet count was 450,000/mm3 or greater. CONCLUSIONS: Platelet count appears to be an independent prognostic factor in renal cell carcinoma. It reflects a cascade of biological events correlated with tumor aggressiveness. This observation opens new perspectives for exploring carcinogenesis mechanisms and tumor progression in renal cell carcinoma. PMID- 16469567 TI - Renal cell carcinoma with nodal metastases in the absence of distant metastatic disease (clinical stage TxN1-2M0): the impact of aggressive surgical resection on patient outcome. AB - PURPOSE: Nodal disease in the setting of metastatic renal cell carcinoma is associated with poor prognosis. However, to our knowledge the biology of nodal metastases in the absence of metastatic disease is unknown. We reviewed our experience with treating this subset of patients with aggressive surgical resection. MATERIALS AND METHODS: A total of 2,643 patients underwent nephrectomy at our institution between 1993 and 2003, including 40 with positive lymph nodes but no systemic metastases. All 40 patients underwent nephrectomy with extended retroperitoneal lymphadenectomy and they are the subjects of this study. Pathological characteristics and clinical outcomes were assessed. RESULTS: Median patient age was 58 years and 62% of the patients were male. Median tumor size was 11 cm. Local stage was T1 in 3% of cases, T2 in 17%, T3a in 30%, T3b in 47% and T4 in 3%. Perinephric fat invasion was present in 77% of patients and positive margins were identified in 17%. Nodal status was N1 in 30% of patients and N2 in 70%, including 10 with masses of matted nodes. Histology was conventional in 63% of cases and papillary in 17%. The remaining 20% of patients had sarcomatoid dedifferentiation. Excluding the 10 patients with matted nodes the median number of nodes harvested per patient was 7 with a median of 2 that were positive. Extranodal extension was present in 70% of cases, while in 70% disease recurred at a median of 4.9 months. Median actuarial disease specific survival was 20.3 months. At a median followup of 17.7 months 30% of patients had no evidence of disease, 8% had disease and 62% had died. On multivariate analysis more than 1 positive node was predictive of decreased recurrence-free survival (HR 2.83, 95% CI 1.06 to 7.61, p = 0.039) and overall survival (HR 9.33, 95% CI 1.85 to 47.09, p = 0.007). CONCLUSIONS: Nodal metastasis with renal cell carcinoma is an independent predictor of prognosis in patients with M0 disease. Even in the absence of distant metastatic disease patients with positive nodes should be targeted for aggressive surgical resection, followed by clinical trials of adjuvant therapy to improve the outcome. PMID- 16469568 TI - Virtual cystoscopy by intravesical instillation of dilute contrast medium: preliminary experience. AB - PURPOSE: We determined whether virtual cystoscopy based on helical data sets can yield urinary capabilities similar to those achieved by fiber-optic cystoscopy. MATERIALS AND METHODS: A total of 11 patients with ultrasound detected bladder tumors underwent pelvic CT as a single volumetric scan after preliminary bladder distention with saline mixed with contrast medium using a 6Fr infant feeding tube. Cystoscopy was simulated based on a 3-dimensional helical CT data set in real time. Source raw CT data for virtual cystoscopy were reconstructed and navigation was done in real time using surface rendering navigation software. These images and findings were then compared with conventional cystoscopy findings. RESULTS: An attenuation gradient of 350 HU between the vesical mucosa and urine was noted. Two of the 14 tumors (11 patients) were missed and each was 0.7 cm. All tumors greater than 0.9 cm were detected. Overall sensitivity was 85.7%. There were no false-positive findings. There was good correlation with tumor location and size. The ureteral orifices and their relationship to the tumor were also well appreciated. Subtle mucosal changes on conventional cystoscopy were not delineated by virtual cystoscopy. CONCLUSIONS: Our method of instilling dilute contrast medium in the bladder offers an excellent attenuation gradient. The overall sensitivity of tumor detection is better than that reported for intravenous contrast medium and pneumocystoscopy. PMID- 16469569 TI - Novel approach of human epidermal growth factor receptor 2 detection in noninvasive and invasive transitional cell carcinoma of the bladder. AB - PURPOSE: In recent years over expression of HER2 has been identified in malignant tumors of organs other than breast. Studies of bladder carcinoma that analyzed HER2 protein expression and gene amplification with a variety of nonstandardized methods have shown heterogeneous results. The results reported vary from 2% to 74% of protein over expression, to 4% to 59% of gene amplification of HER2, possibly due to differences in study design, material selection or laboratory methodology. MATERIALS AND METHODS: In the present study archival tissue from 87 patients with noninvasive papillary (25) and invasive (62) TCC was analyzed for amplification of the HER2 gene and over expression of its encoded protein. HER2 protein expression was detected by immunohistochemistry using the HercepTest. Routinely processed paraffin embedded tissue was investigated for HER2 gene amplification using CISH and FISH. RESULTS: Of the invasive 37 (58%) and of the noninvasive 19 (76%) transitional cell carcinomas investigated showed over expression of the HER2 protein (3+ and 2+) using a standardized immunohistochemical method. HER2 gene amplification assays performed on positive cases evaluated by immunohistochemistry were obtained in 81% and 43% of 3+ and 2+ HER2 protein over expressing invasive, respectively, and in 21% of noninvasive papillary bladder tumors. HER2 gene amplification detection results using CISH and FISH showed a concordance of 100%. The occurrence of aneusomy of chromosome 17 in association with HER2 gene amplification was investigated. CONCLUSIONS: Validation of the HER2 oncogene in bladder cancer may allow for the potential use of Herceptin(R) antibody therapy. Therefore, the appropriate treatment approach has to be based on reliable and standardized analysis. Our results indicate that CISH could provide an accurate and practical alternative to FISH for the clinical diagnosis of HER2 oncogene amplification in bladder cancer. PMID- 16469571 TI - The case for early cystectomy in the treatment of nonmuscle invasive micropapillary bladder carcinoma. AB - PURPOSE: Micropapillary bladder carcinoma is a rare variant of UC. Due to paucity of data regarding treatment outcomes, patients with nonmuscle invasive micropapillary UC often receive intravesical therapy in an attempt at bladder preservation. MATERIALS AND METHODS: We reviewed the records of all patients evaluated at our institution who had micropapillary UC of the bladder. Of these, 44 had nonmuscle invasive disease at presentation and form the basis of this report. RESULTS: Mean patient age was 64.3 years (range 45 to 81) with a male-to female ratio of 13:1. Stage distribution at presentation was 5 Ta (11%), 4 CIS (9%) and 35 T1 (80%). Median CSS was 81 months. Kaplan-Meier estimates of 5 and 10-year CSS rates were 64% and 26%, respectively. Intravesical BCG therapy was attempted in 27 patients (61%). Of these 27 patients, 67% (18 of 27) had progression (cT2 or greater), including 22% in whom metastatic disease developed. Only 19% of patients (5 of 27, all T1) remain disease-free with an intact bladder at a median followup of 30 months. A total of 30 patients (68%) underwent cystectomy. Among patients who underwent cystectomy after progression (18), median CSS was 61.7 months with no patient surviving 10 years, whereas among those undergoing cystectomy as initial therapy (12), median survival was not reached and the 10-year CSS rate was 72%. CONCLUSIONS: Intravesical BCG therapy appears to be ineffective against micropapillary UC. Our results suggest that the optimal treatment strategy for nonmuscle invasive micropapillary UC is radical cystectomy performed before progression. PMID- 16469572 TI - A critical analysis of perioperative mortality from radical cystectomy. AB - PURPOSE: Operative mortality from radical cystectomy has decreased as a result of improvements in surgical and anesthetic care. We reviewed the perioperative deaths from a large group of patients treated with radical cystectomy for primary bladder cancer. MATERIALS AND METHODS: All perioperative mortalities from radical cystectomy were identified from a single high volume institution. The medical records were reviewed to assess the cause of death as well as possible contributing factors. RESULTS: From August 1971 to December 2001, 1,359 patients with primary bladder cancer were treated with radical cystectomy and pelvic iliac lymphadenectomy at our institution. Of these patients, 27 (2%) died within 30 days of surgery or before discharge from hospital. Median patient age at surgery was 67 years (range 47 to 78) and males accounted for 81% of the patients. The median time to death was 28 days from cystectomy (range 0 to 80). Most deaths were cardiovascular related (including acute myocardial infarction, cerebrovascular accident, arterial thrombosis) or due to septic complications with resulting multi-organ system failure, followed by pulmonary embolism, hepatic failure and hemorrhage. Septic related mortality was most often associated with postoperative urine or bowel leak. While most deaths occurred before hospital discharge, 2 patients died at home due to a late pulmonary embolus. No association was seen between pathological stage or type of urinary diversion and mortality. CONCLUSIONS: Perioperative mortality from radical cystectomy is low in this group of patients. Most deaths are due to cardiovascular or septic complications. Careful patient selection and meticulous surgical technique may help decrease the incidence of perioperative mortality. PMID- 16469574 TI - The detection and potential economic value of complexed prostate specific antigen as a first line test. AB - PURPOSE: Prostate cancer detection is subject to a number of variables that can lead to unnecessary biopsies and associated costs. Measuring cPSA has been proposed as an alternative to tPSA for the early detection of prostate cancer. MATERIALS AND METHODS: Between November 1998 and April 2000, 1,362 men underwent transrectal ultrasound guided biopsies at 7 institutions. Of 1,243 evaluable men 467 with tPSA between 2.5 and 6.0 ng/ml, and normal digital rectal examination were analyzed. Statistical analysis used to compare cancer detection rates between PSA assays was performed using the Mann-Whitney U test. A separate group of 2,807 men who participated in a free cancer detection program was used to determine the current tPSA distribution and assess the economic impact of cPSA. RESULTS: Cancer was detected in 31.5% of the men (147 of 467) with tPSA between 2.5 and 6.0 ng/ml. Using a 2.2 ng/ml cPSA cutoff point detected 93.9% of cancers and would have avoided 20.3% of unnecessary biopsies in men with tPSA between 2.5 and 4.0 ng/ml. A 2.2 ng/ml cPSA cutoff point achieved an 11.9% overall decrease in the number of unnecessary biopsies in the tPSA range of 2.5 to 6.0 ng/ml with accompanying 98% sensitivity. The decrease in unnecessary biopsies is potentially associated with substantial health care cost savings. CONCLUSIONS: In the clinically relevant sensitivity ranges a 2.2 ng/ml cPSA cutoff point decreases the number of unnecessary biopsies and maintains higher specificity than a tPSA threshold of 2.5 ng/ml, illustrating the potential value of cPSA as a first line diagnostic test. PMID- 16469576 TI - Pathological characteristics of prostate cancer detected through prostate specific antigen based screening. AB - PURPOSE: Since the introduction of PSA testing for CaP, there has been an increase in CaP detection. However, it is uncertain to what extent clinically insignificant tumors are being diagnosed and treated. In a large, community based population we determined the pathological characteristics of screening detected cancers. MATERIALS AND METHODS: From 1989 to 2001, 35,661 men were enrolled in a longitudinal prostate cancer screening study. Data were available on 3,492 of the 3,568 men (98%) diagnosed with CaP during this study period. Radical prostatectomy was performed in 2,254 men (63%). Clinical stage, Gleason score and pathological analysis were recorded and analyzed in the context of preoperative PSA, digital rectal examination findings, PSA velocity and the year of cancer detection. RESULTS: CaP was detected in 10% of men. Virtually all cases were clinically localized (99.8%) and approximately 70% treated with radical prostatectomy were pathologically organ confined disease. Fewer than 10% of tumors would be considered clinically insignificant based on 2 previously published pathological criteria. CONCLUSIONS: Compared to the high prevalence of CaP found in autopsy studies there is a lower detection rate using current screening protocols. Although the outcomes are unknown if these tumors had been left untreated, the majority met pathological criteria for significant cancer. PMID- 16469577 TI - Evidence to support a continued stage migration and decrease in prostate cancer specific mortality. AB - PURPOSE: We evaluated whether the proportion of patients with a postoperative PSA DT less than 3 months, a surrogate for PCSM, decreased significantly during the PSA era. MATERIALS AND METHODS: Between July 1988 and July 2002, 3,719 men with clinically localized prostate cancer treated with RP comprised the study cohort. A chi-square metric was used to compare the preoperative and postoperative characteristics, 5-year actual PSA failure rates, and PSA-DTs for patients treated during the 2 equally divided eras of the early PSA era, July 1988 to July 1995 and the late PSA era, August 1995 to July 2002. RESULTS: Patients presenting in the more recent PSA era were of younger age (p < 0.0001), with earlier stage (p < 0.0001) and lower grade disease (p = 0.01). Similarly, patients had lower grade (p < 0.001), stage (p < 0.0001), and positive margin (p < 0.0001) and lymph node rates (p = 0.0002) at RP. The 5-year actual PSA failure rates decreased from 14.3% in the early PSA era to 2.5% in the later PSA era (p < 0.0001). There was a 37% reduction in the proportion of patients with a PSA-DT less than 3 months, corresponding to a decrease in absolute magnitude from 9% to 5.7% between the 2 eras. Absolute reductions of 3.1% and 9% were also noted for the proportion of PSA-DTs of 3 to 5.99 months and 6 to 11.99 months, respectively, whereas PSA-DTs of 12 months or greater increased by 15.3%. CONCLUSIONS: During the recent PSA era, postoperative PSA failure has significantly decreased and PSA-DTs have increased, suggesting that PCSM will continue to decrease. PMID- 16469578 TI - Prostate specific antigen remains an independent predictor of cancer at prostate biopsy in black american men but not in white men: results from a consecutive series of 914 men. AB - PURPOSE: Black American men may be at increased risk for prostate cancer but differences in prebiopsy parameters between black and white men have not been fully examined. Therefore, we identified the prebiopsy parameters that may be predictive of prostate cancer in black and white men. MATERIALS AND METHODS: From January 2000 to July 2004, 914 consecutive men undergoing prostate needle biopsy were prospectively examined by a single urologist. Urinary symptoms were measured by AUASS. Prebiopsy parameters recorded were PSA, free PSA, DRE, age, race, biopsy history, prostate volume, TRUS lesion and AUASS. RESULTS: Prostate biopsy was performed in 914 men with a mean age of 63.9 years. Mean PSA in the entire cohort was 11.2 ng/ml (median 5.8). Abnormal TRUS and abnormal DRE were found in 37% and 52% of men, respectively. Mean AUASS was 9.6. The overall positive biopsy rate was 37%. In black and white men the positive biopsy rate was 44% and 34%, respectively. Analysis of AUASS indicated that 47% of patients had low symptom scores (less than 7), 39% had moderate scores (8 to 19) and 14% had severe scores (20 to 35). Multivariate analysis revealed that PSA was an independent predictor of positive biopsy in black but not in white men (p = 0.001 and 0.79, respectively). Multivariate analysis also showed that race alone was an independent predictor of positive prostate biopsy (p = 0.013). CONCLUSIONS: PSA remains an independent predictor of positive prostate biopsy on multivariate analysis. Other independent predictors are black race, age, low AUASS, prostate volume, abnormal DRE, no previous biopsy and abnormal TRUS. In the black group low AUASS, PSA, no previous biopsy and DRE were unique independent predictors, while in the white group age and abnormal TRUS were unique predictors. PMID- 16469580 TI - Classification and regression tree analysis for the prediction of aggressive prostate cancer on biopsy. AB - PURPOSE: Prostate cancer screening allows early cancer detection but not all patients benefit from subsequent therapy. Thus, identifying patients who are likely to harbor aggressive cancer could significantly decrease the number of prostate biopsies performed. MATERIALS AND METHODS: Data were collected on 1,563 consecutive referred men with serum PSA 10 ng/ml or less who underwent an initial prostate biopsy. Predictors of aggressive cancer (Gleason sum 7 or greater) were identified using CART analysis. Model building was done in a randomly selected training set (70% of the data) and validation was completed using the remaining data. RESULTS: Cancer was detected in 406 men (26.1%). Gleason 7 or greater cancer was found in 130 men (8.3%). CART created a decision tree that identified certain groups at risk for aggressive cancer, namely 1) PSAD greater than 0.165 ng/ml/cc, and 2) PSAD greater than 0.058 to 0.165 ng/ml/cc or less, age greater than 57.5 years and prostate volume greater than 22.7 cc. The incidence of aggressive prostate cancer was 1.1% when PSAD was 0.058 ng/ml/cc or less in the validation set. The sensitivity and specificity of CART for identifying men with aggressive cancer were 100% and 31.8% for model building data, and 91.5% and 33.5% for the validation data set, respectively. CONCLUSIONS: CART identified groups at risk for aggressive prostate cancer. Application of this CART could decrease unnecessary biopsies by 33.5% when only a diagnosis of high grade prostate cancer would lead to subsequent therapy. PMID- 16469581 TI - Prostate cancer diagnosed after repeat biopsies have a favorable pathological outcome but similar recurrence rate. AB - PURPOSE: We investigated whether repeat prostate biopsies are associated with more favorable prognoses, less extensive disease or higher rates of IC in patients who are ultimately diagnosed with prostate cancer and treated with RRP. MATERIALS AND METHODS: We examined standard clinical and pathological data on 1,357 patients treated with RRP from 1983 to 2001. In addition, we noted the rate of IC in a subgroup of 847 patients in whom tumor volume was measured. RESULTS: Cancer was found in 1,042 patients (77%) at the first biopsy, in 227 (17%) at the second biopsy, in 59 (4%) at the third biopsy and in 29 (2%) at the fourth or later biopsy. Patients with 2 or greater biopsies had a higher rate of clinical T1c stage cancer and larger prostates than patients with only 1 biopsy (each p < 0.0001). After RRP patients with 1 biopsy had a lower rate of organ confined tumors (61% vs 75%, p < 0.0001), and a higher rate of extracapsular extension, seminal vesicle invasion, lymph node metastases and Gleason sum 7 or greater than other patients. IC was found in 10% of patients with 1 biopsy and 18% of those with 2 or greater biopsies (p = 0.018). Despite these more favorable pathological outcomes there was no difference in biochemical recurrence rate. CONCLUSIONS: Although we found that a greater number of biopsies was related to a better pathological outcome after RRP, the number of biopsies did not predict disease recurrence. The increasing number of biopsies currently being performed, especially in patients with larger prostates, likely results in higher rates of IC. PMID- 16469583 TI - Significance of atypical and suspicious small acinar proliferations, and high grade prostatic intraepithelial neoplasia on prostate biopsy: implications for cancer detection and biopsy strategy. AB - PURPOSE: We reviewed our results from a urological pathology reference laboratory with respect to the incidence of HGPIN, and atypical and suspicious lesions in the spectrum of ASAP. Subsequent CaP findings on repeat biopsy with relevant clinical implications were assessed. MATERIALS AND METHODS: A review of 42,667 prostate biopsies was performed. We defined atypical and suspicious as variants of ASAP with suspicious being more worrisome for CaP. Findings were correlated with the location of CaP on repeat prostate biopsy. RESULTS: The rate of subsequent CaP detection was significantly higher for an initial diagnosis of suspicious findings (51% or 54 of 107 cases) than for atypical findings (34% or 39 of 116) or HGPIN (22% or 79 of 358, p < 0.001). CaP was found on the same side of the prostate in 61 of 78 (78%), 30 of 39 (77%) and 41 of 54 patients (76%) with initial HGPIN, atypical and suspicious biopsies, respectively. There was no significant difference among the 3 groups in the likelihood of future CaP at the same site or the same side of the prostate. CONCLUSIONS: Patients with a suspicious biopsy were significantly more likely to have CaP on future biopsy than those with atypical findings or HGPIN, suggesting that there may be divisions with prognostic significance in the larger category of ASAP. To our knowledge the reproducibility of recognizing such divisions remains to be established. Neither atypical nor suspicious lesions were more likely than HGPIN to predict CaP at the same site or side of the prostate as the original diagnosis. Repeat biopsy may be indicated in any patient with HGPIN, or atypical or suspicious lesions and this biopsy should not be limited to the site or side of the original pathological findings. PMID- 16469585 TI - Do the benefits of finasteride outweigh the risks in the prostate cancer prevention trial? AB - PURPOSE: The Prostate Cancer Prevention Trial demonstrated that finasteride could reduce the incidence of prostate cancer by 25%. However, its use was also associated with an increased risk of high grade cancer resulting in uncertainty surrounding the net benefits of therapy. MATERIALS AND METHODS: We used the Montreal Prostate Cancer Model, a validated Markov model of prostate cancer progression, to compare the forecasted survival in treated and untreated men. The conditions of the model were varied to reflect different assumptions about whether the cancer grade difference observed in the PCPT was real or a treatment associated artifact, and whether cancers detected on end of study biopsies were clinically significant. RESULTS: For a hypothetical cohort of 1,000, 62-year-old men treated with finasteride, an increased survival of 140 life-years (0.14 years per individual) is forecasted if all diagnosed cancers are considered. If tumor grade differences are held to be artifactual, the forecasted benefits increase to 200 life-years. However, if the tumor grade difference is real and only clinically detected cancers are considered, estimated increased survival is only 20 life-years (0.02 years per individual). CONCLUSIONS: The primary prevention of prostate cancer with finasteride looks promising. However, at the present time it should only be considered with caution until we have answered critical questions surrounding the difference in cancer grade observed in the PCPT and the clinical significance of cancers detected on protocol directed end of study biopsies. PMID- 16469587 TI - Validation of a nomogram for prediction of side specific extracapsular extension at radical prostatectomy. AB - PURPOSE: We have previously have reported a tree structured regression model for predicting SS-ECE. Others recently reported a logistic regression based SS-ECE nomogram. We developed a nomogram and compared the performance and discriminant properties of the tree regression and the nomogram in a contemporary cohort of European patients treated with radical retropubic prostatectomy. MATERIALS AND METHODS: The cohort consisted of 1,118 patients with pretreatment prostate specific antigen 0.1 to 73.2 ng/ml (median 6.6). Each of the 2,236 prostate lobes was considered separately. Clinical stage, pretreatment PSA, biopsy Gleason sum, percent positive cores and percent cancer in the biopsy specimen were used as predictors in a logistic regression model predicting SS-ECE. Regression coefficients were then used to generate an SS-ECE nomogram. Performance characteristics and discriminant properties of the previously published tree regression were also tested in the same cohort. For internal validation and to decrease overfit bias 200 bootstrap re-samples were applied to accuracy estimates for each method. RESULTS: ECE was present in 303 of 1,118 radical retropubic prostatectomy specimens (27%) and in 385 lobes (17%). In logistic regression models all variables were statistically significant multivariate predictors of SS ECE except the percent of positive biopsy cores (p = 0.7). Bootstrap corrected predictive accuracy of the SS-ECE nomogram was 0.840 vs 0.700 for the tree regression model. CONCLUSIONS: Logistic regression based nomogram predictions of SS-ECE are highly accurate and represent a valuable aid for assessing the risk of ECE prior to surgery. PMID- 16469589 TI - Robotic extraperitoneal radical prostatectomy: an alternative approach. AB - PURPOSE: Laparoscopic radical prostatectomy with or without a robot has been increasingly performed worldwide, primarily using a transperitoneal approach. We report our experience with daVinci(R) robot assisted extraperitoneal laparoscopic radical prostatectomy. MATERIALS AND METHODS: A total of 325 patients underwent robot assisted extraperitoneal laparoscopic radical prostatectomy for clinically localized prostate cancer at our center during a 2-year period. Perioperative data, and oncological and functional results were prospectively recorded. RESULTS: Perioperative demographics included mean age, PSA and Gleason score, which were 60 years (range 42 to 76), 6.6 ng/ml (range 0.6 to 26) and 6 (range 5 to 9), respectively. Preoperative clinical stage was 81%, 16% and 3% for T1c, T2a and T2b, respectively. Average total operative time was 130 minutes (range 80 to 480). Intraoperative data included a mean blood loss of 196 cc with no open conversions. Bilateral, unilateral and nonnerve sparing prostatectomy was performed in 70%, 24% and 6% of patients, respectively. Of the patients 96% were discharged home within 8 to 23 hours of surgery. Pathological stage was pT2a, pT2b, pT3a and pT3b in 18%, 63%, 14% and 5% of all radical prostatectomy specimens, respectively, with an overall positive surgical margin rate of 13%. Two of 92 patients had positive nodal disease after lymph node dissection. Continence and erectile function were measured. CONCLUSIONS: The extraperitoneal approach offers the advantages of improved dexterity and visualization of the robot, while avoiding the abdominal cavity and potential associated morbidity. As surgeons gain more experience with this new technology, the extraperitoneal approach simulating the standard open retropubic technique is likely to gain popularity. PMID- 16469591 TI - Impact of patient age at treatment on outcome following radical retropubic prostatectomy for prostate cancer. AB - PURPOSE: Historically young patients with prostate cancer have been found to have poorer outcomes. Recent studies suggest favorable pathological findings and improved survival in younger patients undergoing RRP. We assessed age at treatment as a predictor of post-RRP survival. MATERIALS AND METHODS: We identified 5,509 patients treated with RRP for prostate cancer at our institution between 1987 and 1995. Age at treatment was classified into categories of younger than 55, 55 to 59, 60 to 64, 65 to 69 and 70 years or older. CSS, sPFS and biochemical PFS were estimated by the Kaplan-Meier method and analyzed using Cox proportional hazard models. RESULTS: Younger patients had lower preoperative prostate specific antigen, and tumor grade and stage. CSS, sPFS and biochemical PFS were similar across age groups but overall survival decreased with older age at treatment. After multivariate adjustment the risk of cancer death was lower in patients 70 years or older (RR 0.53, 95% CI 0.30 to 0.90), while the risk of progression was lower in all age groups compared to that in men younger than 55 years (RR 0.57 to 0.62). On stratified subset analysis sPFS was progressively worse with younger age in patients with high risk pathological findings. However, the addition of age to multivariate models incorporating preoperative prostate specific antigen, pathological features and adjuvant therapy failed to improve their predictive value for CSS and sPFS. CONCLUSIONS: Despite more favorable clinicopathological features younger patients undergoing RRP for prostate cancer have survival similar to that of older counterparts. Given the greater proportionate impact of prostate cancer on survival, it is particularly important to pursue aggressive treatment in younger patients. PMID- 16469592 TI - Predictors of symptom severity in patients with chronic prostatitis and interstitial cystitis. AB - PURPOSE: Numerous studies have been performed to identify potential risk factors for CP/CPPS and IC. However, few studies have been done to identify predictors of disease severity. MATERIALS AND METHODS: A total of 174 men with CP/CPPS and 111 women with IC completed questionnaires to quantify symptom severity and identify demographic, medical and psychosocial characteristics. Symptom severity was assessed with the National Institutes of Health CPSI in men, and the O'Leary-Sant ICSI and problem index in women. Univariate and multivariate analyses were performed to identify characteristics predictive of worse symptoms. RESULTS: The mean National Institutes of Health CPSI score in men was 15.32, and the mean O'Leary-Sant ICSI and problem index in women was 19.17. The most commonly reported comorbidities were allergies, sinusitis, erectile dysfunction and irritable bowel syndrome in men, and allergies, urinary incontinence, sinusitis and irritable bowel syndrome in women. In the 2 sexes self-reported urinary frequency and urgency, worse depression scores and lower education level were independent predictors of worse symptom severity. In men additional independent predictors were self-reported pelvic pain, fibromyalgia and previous heart attack, and in women an additional independent predictor was postmenopausal status. CONCLUSIONS: There are several common medical conditions associated with urological pelvic pain syndromes in men and women. Few of them were predictive of symptoms severity in this analysis. Self-reported pelvic pain symptoms, education and depression severity were the factors most strongly predictive of symptom severity in patients with CP/CPPS and IC. PMID- 16469594 TI - Renal and extrarenal predictors of nephrectomy from the national trauma data bank. AB - PURPOSE: The kidney is injured in 1.4% to 3.0% of all trauma cases. The management of renal injuries is controversial, as reflected in regional and institutional variations in treatment preferences. Using a national trauma database we identified independent risk factors for nephrectomy. MATERIALS AND METHODS: The population was selected from the National Trauma Data Bank, a voluntary data repository containing all trauma admissions to 268 participating trauma centers. Patients with renal injuries were identified by Abbreviated Injury Scale codes. Patient demographic, associated injuries and facility characteristics were recorded. Univariate and Poisson regression analysis with clustering by facility was performed. RESULTS: Renal injury was present in 8,465 patients. Nephrectomy was performed in 4% of all blunt and 21% of all cases of penetrating renal injuries. Only 0.5% of blunt renal injury cases underwent repair compared with 15% of those of penetrating injuries. On multivariate analysis renal injury severity was the strongest predictor of nephrectomy. The relative risk of nephrectomy for grade V renal injuries was 146 (95% CI 74 to 289) and 33 (95% CI 13 to 89) in the blunt and penetrating models, respectively. The need for laparotomy and surgery on other intra-abdominal organs predicted nephrectomy in patients with blunt and penetrating injuries. Hospital trauma designation did not statistically impact nephrectomy rates. CONCLUSIONS: The severity of renal injury based on the AAST organ injury scale for Renal Trauma is the strongest risk factor for nephrectomy. The need for surgery on other intra abdominal injuries increases the risk of nephrectomy to a lesser extent. In cases of blunt trauma severe renal injury usually necessitates nephrectomy. PMID- 16469596 TI - Technique and results of urethroplasty for female stricture disease. AB - PURPOSE: Urethral stricture disease in females is uncommon and is often treated with repeat dilation or internal urethrotomies. Various surgical techniques to repair strictures have been described with successful results. However, these techniques are cumbersome to use. The vaginal inlay flap is simple and easy to learn. To our knowledge this is the first report of its use and clinical results in a series of patients from 2 institutions. MATERIALS AND METHODS: Eight symptomatic women with a history of traumatic or difficult catheterization, a history of at least 1 urethral dilation or urethrotomy and difficult or a failed attempt at catheter placement underwent urethroplasty. The technique consisted of incising the posterior aspect of the stricture and advancing a vaginal inlay flap. A retrospective chart review was performed. RESULTS: Followup was 1 to 9 years. All patients had subjective relief of symptoms and could easily catheterize with a 14Fr catheter. Average caliber of the urethra increased from 9.25Fr to 16.5Fr and post-void residual urine decreased from 130 to 15 cc. One patient with a hypotonic bladder was in retention, which resolved during 3 months. One patient underwent repeat dilation once 3 weeks after the primary procedure with no recurrence. No patient had stress urinary incontinence. There were no immediate or delayed serious complications. CONCLUSIONS: Urethral stricture disease in females is an uncommon entity that can cause voiding symptoms, recurrent infections, retention and renal impairment. This method of surgical repair offers a durable result and has a low incidence of complications. PMID- 16469598 TI - Feminizing reconstructive surgery for ambiguous genitalia: the Leipzig experience. AB - PURPOSE: We report the results of feminizing reconstructive surgery for ambiguous genitalia with regard to the degree of virilization according to Prader and the long-term outcome with special emphasis on sexual intercourse. MATERIALS AND METHODS: Patients 16 to 46 years old with CAH (41) and MPH (17) were followed continuously by an interdisciplinary team that provided standardized hormone substitution and reconstructive surgery. More virilized genitalia were treated using a 2-stage procedure. RESULTS: Of patients with CAH no surgery and clitoris reduction were done in 4 and 2 (Prader I and II), no surgery, a 1-stage and a 2 stage procedure were done in 2, 3 and 4 (Prader III), a 2-stage procedure, a 1 stage procedure and surgery for fistula were done in 16, 4 and 2, respectively, while 1 refused surgery (Prader IV), and a 2-stage procedure was done in 5, including pull-through vaginoplasty in 2 (Prader V). Of patients with MPH no surgery was done in 6 (Prader I and II), a 1-stage procedure and a 2-stage procedure were done in 3 and 1 (Prader III), respectively, and a 2-stage procedure was done in 6, while 1 refused surgery (Prader IV). Revision vaginoplasties were necessary in 7 patients (12.1%) because of scar stenosis in those who underwent 1-stage as well as 2-stage procedures. None of our patients required reconstructive surgery to create a neovagina. Of the 58 patients 32 achieved sexual intercourse and in 17 the postoperative result would allow sexual intercourse, while in 3 the possibility of sexual intercourse is uncertain. For 3 patients sexual intercourse would not be possible since feminizing reconstructive surgery was refused. One patient could not be followed. CONCLUSIONS: Two-stage surgery leads to a favorable outcome in patients with CAH and MPH, and virilized genitalia (Prader IV and V). PMID- 16469599 TI - Childhood urinary symptoms predict adult overactive bladder symptoms. AB - PURPOSE: A relationship between childhood urinary symptoms and adult lower urinary tract symptoms in women is often clinically suspected. In this analysis we investigated the relationship between childhood and adult urinary symptoms in middle-aged women. MATERIALS AND METHODS: A population based cohort of 2,109 women 40 to 69 years old who were members of a large health maintenance organization was randomly selected from age and race strata. Through self reported questionnaires, women recalled a childhood history of and current urinary lower urinary tract symptoms, including frequent daytime urination, nocturia, urinary incontinence, nocturnal enuresis and UTIs. Current incontinence was also classified as urge or stress incontinence. Multivariate analysis was used to evaluate the association between childhood and current lower urinary tract symptoms controlling for age, race, hysterectomy status, parity, oral estrogen use, body mass index and diabetes. RESULTS: Women who reported childhood daytime frequency were more likely to report adult urgency (OR 1.9, 95% CI 1.3 2.6, p < 0.001). Frequent nocturia in childhood was strongly associated with adult nocturia (OR 2.3, 95% CI 1.5-3.5, p < 0.001). Childhood daytime incontinence was associated with adult urge incontinence (OR 2.6, 95% CI 1.1-5.9, p < 0.05), as was childhood nocturnal enuresis (OR 2.7, CI 1.3-5.5, p < 0.01). A history of more than 1 childhood UTI was associated with adult UTIs (OR 2.6, 95% CI 1.5-4.5, p < 0.001). CONCLUSIONS: Childhood urinary symptoms and UTIs were significantly associated with adult overactive bladder symptoms. There is a need to investigate the significance of childhood symptoms as predictors of eventual adult disorders to determine whether treatment of childhood symptoms will alter the prevalence of eventual adult disorders, and if such a history should alter clinical care of the older adult with OAB symptoms. PMID- 16469600 TI - Brain activation during detrusor overactivity in patients with Parkinson's disease: a positron emission tomography study. AB - PURPOSE: Patients with Parkinson's disease often have urine storage symptoms, such as urinary urgency, frequency and incontinence, which are induced by detrusor overactivity. However, little is known of the mechanisms inducing detrusor overactivity in this disease. We have previously examined the human brain response to bladder filling in healthy male volunteers using positron emission tomography. We hypothesized that brain activation patterns in response to bladder filling would be different in patients with Parkinson's disease. MATERIALS AND METHODS: Nine male patients with Parkinson's disease were catheterized via the urethra for bladder filling and intravesical pressure monitoring. We performed positron emission tomography, consisting of tasks 1 and 2. For task 1 the bladder was maintained empty via the urethral catheter. For task 2 room temperature water was dripped for bladder filling until the onset of detrusor overactivity. Data acquisition for task 2 was done during detrusor overactivity. Data on each scan were summed on a computer and further analyzed using a statistical parametric mapping procedure. RESULTS: Significant brain activation during detrusor overactivity was found in the periaqueductal gray, supplementary motor area, cerebellar vermis, insula, putamen and thalamus. The most prominent activation was found in the cerebellum. The pons was not activated during detrusor overactivity. CONCLUSIONS: Alteration in brain activation sites in response to bladder filling may be related to the pathophysiology of detrusor overactivity in patients with Parkinson's disease. PMID- 16469603 TI - Sacral neuromodulation in women with idiopathic detrusor overactivity incontinence: decreased overactivity but unchanged bladder contraction strength and urethral resistance during voiding. AB - PURPOSE: We evaluated the effect of sacral (S3) nerve neuromodulation on voiding in women with idiopathic detrusor overactivity incontinence. MATERIALS AND METHODS: Urodynamic measurements in all patients implanted in 1990 to 2003 were reconsidered. Patients were included if these measurements, which were done at baseline and after 6 months, could be analyzed completely and reliably. Maximum detrusor pressure, amplitude of the highest involuntary detrusor contraction and end fill volume were used as parameters characterizing the degree of detrusor overactivity. Urethral resistance and bladder contraction strength during voiding were characterized by the bladder outlet obstruction index, the urethral resistance factor, average pressure, the slope of the low pressure side of the pressure flow plot, the bladder contractility index and the bladder contraction strength parameter. RESULTS: A total of 33 women were included. Detrusor overactivity parameters were significantly improved at followup. In addition, the supine position of the patient during filling in followup measurements proved less provocative with respect to overactivity than the standing position in the majority of measurements at baseline. Consequently bladder volumes at which voiding was initiated were considerably higher at followup. Changes in the parameters characterizing urethral resistance and bladder contraction strength during voiding were not unambiguous. However, exactly those parameters that appeared volume independent in a previous study were not significantly different. CONCLUSIONS: Our study confirmed the depressant effect of sacral (S3) nerve neuromodulation on detrusor overactivity. No effect on urethral resistance and bladder contraction strength during voiding could be demonstrated using volume independent parameters. PMID- 16469601 TI - Safety and tolerability of tolterodine for the treatment of overactive bladder in men with bladder outlet obstruction. AB - PURPOSE: Antimuscarinic therapy for men with OAB and BOO is perceived as a potential risk for urinary retention. Using pressure flow urodynamics, we evaluated the safety of tolterodine vs placebo in men with OAB and BOO. MATERIALS AND METHODS: Men (older than 40 years) with BOO and confirmed detrusor overactivity were randomized to tolterodine (2 mg twice daily in 149) or placebo (in 72) for 12 weeks. Primary end points were Qmax and pdetQmax. RESULTS: Median treatment differences in Qmax (-0.7 ml per second, 95% CI -1.6 to 0.4) and pdetQmax (-7 cm H2O, 95% CI -3 to 11) were comparable. Tolterodine significantly reduced the BOOI vs placebo (-9 vs 0, p < 0.02). There were significant treatment differences in volume to first detrusor contraction (+59 ml, 95% CI 19-100) and maximum cystometric capacity (+67 ml, 95% CI 35-103), favoring tolterodine over placebo (p < 0.003). Change in PVR was significantly greater among patients treated with tolterodine (+25 ml) than placebo (0 ml, p < 0.004). There were no significant between-group differences in the incidence of adverse events. Urinary retention was reported by 1 patient treated with placebo. CONCLUSIONS: Tolterodine did not adversely affect urinary function in men with OAB and BOO. Urinary flow rate was unaltered, and there was no evidence of clinically meaningful changes in voiding pressure and PVR or urinary retention. Tolterodine was well tolerated. These results suggest that antimuscarinics can be safely administered in men with BOO. PMID- 16469605 TI - Recurrent pelvic floor defects after abdominal sacral colpopexy. AB - PURPOSE: The incidence of site specific pelvic organ prolapse defects following sacral colpopexy is not clearly reported. We evaluated site specific pelvic organ defects after colpopexy and determined its impact on patient satisfaction. MATERIALS AND METHODS: A total of 40 women with vault prolapse underwent abdominal sacral colpopexy, culdeplasty and paravaginal repair. Followup consisted of pelvic examination and satisfaction assessment every 6 months. The Baden-Walker classification was used and prolapse halfway to the introitus (grade II) or greater was considered significant prolapse. Surgical failure was identified as grade III prolapse or greater. Satisfaction was assessed on a scale of 1 to 3 with 3 being highly satisfied and according to whether patients perceived a successful outcome. RESULTS: A total of 40 patients with an average age of 66.5 years (range 48 to 81) had an average followup of 25.5 months (range 18 to 42). Of the 40 patients 22 (55%) did not have significant prolapse, including 14 with no prolapse, and 8 with grade I cystocele and/or rectocele. Of the 40 patients 18 (45%) had recurrent significant prolapse, including cystocele in 8 (grades II and III in 4 each), rectocele in 6 (grades II and III in 2 and 4, respectively), and grade II cystocele and rectocele in 3. There was 1 case of recurrent vault prolapse. Eight of 40 cases (20%) were considered surgical failures. Patients without prolapse were highly satisfied (average score 2.95) and 100% considered surgery to have been successful. The recurrent prolapse group was less satisfied (mean score 2.5) and 66.7% considered the surgery successful. CONCLUSIONS: Recurrent pelvic organ prolapse is not an uncommon finding after colpopexy and it may adversely affect patient satisfaction. PMID- 16469607 TI - Transobturator versus transabdominal mid urethral slings: a multi-institutional comparison of obstructive voiding complications. AB - PURPOSE: In the last year TO slings have become an increasingly popular alternative to TA slings for the surgical treatment of SUI. Proposed advantages of the transobturator approach include improved speed, safety and the reduction of obstructive complications. We assessed outcomes of TO and TA slings in a large series of women treated at several institutions to compare the rate of obstructive complications from these procedures. MATERIALS AND METHODS: We reviewed the charts of 504 consecutive women who had synthetic mid urethral sling procedures (154 TO or 350 TA) performed by 24 different urologists for SUI at 8 institutions from 2002 to 2004. Obstructive complications were defined as increased PVR (greater than 100 cc), or the need for CIC, prolonged Foley catheter drainage or urethrolysis. RESULTS: While TO and TA sling procedures appeared to be similarly efficacious in eliminating the need for incontinence pad use (TO 89%, TA 86%, p = 0.36), the transobturator approach was associated with fewer obstructive complications (TO 11.0%, TA 18.3%, p < 0.05). Urethrolysis was required in none of the 154 TO cases and 8 of 350 (2.3%) TA cases. Concomitant pelvic surgery did not significantly increase the likelihood of obstructive voiding complications in either group. CONCLUSIONS: Although TO and TA sling procedures had similar short-term results for decreasing pad use in patients with stress urinary incontinence, the transobturator approach is associated with fewer obstructive voiding complications. PMID- 16469608 TI - Tracking of longitudinal changes in measures of benign prostatic hyperplasia in a population based cohort. AB - PURPOSE: We characterized how longitudinal changes in PSA, prostate size, maximum urinary flow rates and lower urinary tract symptoms track together over time. MATERIALS AND METHODS: In 1990, 2,115 white men, randomly selected from the Olmsted County, Minnesota population, completed validated questionnaires during a home visit by a study assistant. A 25% random subsample underwent clinical evaluation including transrectal ultrasonography, serum PSA and assessment of maximum urinary flow rates. Examinations and questionnaires were repeated biennially through 2002. Longitudinal changes in these measurements were estimated with 2-stage models. Annualized changes were correlated and also dichotomized at various percentiles and examined in age adjusted logistic regression models predicting symptom increases in the upper 80th percentile. RESULTS: Correlations between changes in prostate volume, PSA levels, maximum flow rates and urinary symptoms were modest (age adjusted Spearman correlation coefficients: volume and symptoms 0.08, p = 0.06; PSA and symptoms 0.06, p = 0.20; maximum flow rate and symptoms -0.08, p = 0.05). However, PSA and prostate volume annual increases in the upper 80th percentile were each associated with an approximately 2-fold increased risk of symptom changes in the upper 80th percentile. As PSA and prostate volume changes increased from the 50th to the 90th percentiles, the odds of having symptoms in the upper 80th percentile also increased. CONCLUSIONS: While overall correlations among changes in each of these variables were modest, men with more rapid annual increases in PSA levels and prostate volumes were also likely to have more rapid increases in urinary symptoms. PMID- 16469610 TI - Geographic variations in the use of medical and surgical therapies for benign prostatic hyperplasia. AB - PURPOSE: Patients with BPH have several treatment options. Little is known about geographic variations in surgical rates for BPH and the market relationships to medical therapy, health resources and sociodemographic factors. MATERIALS AND METHODS: We conducted a cross-sectional study using administrative data from 5 states in 2000. Rates of surgical and medical therapy were calculated per 100,000 men 55 years old or older. Main outcome measures were county level weighted coefficient of variation and systematic component of variation in therapy rates, as well as surgery rates as a function of medication dispensing rates, health care resources and sociodemographic characteristics. RESULTS: North Carolina had the lowest surgery rates (26.3 minimally invasive procedures and 332.1 invasive surgeries per 100,000) and finasteride dispensing rates (503.5 per 100,000). Overall rates of medical therapy were 5 times higher than surgery rates. Geographic variations in surgical and medical therapy rates were significant for each state, and North Carolina had the greatest variation. An increase of 11.6 per 100,000 (95% CI, 6.5-55.8) in annual county level finasteride dispensing would be associated with a decrease in the surgery rate of 1 per 100,000, controlling for other variables. CONCLUSIONS: There is significant systematic variation in rates of surgical and medical therapy for BPH at county and state levels. The relationship between finasteride and surgery in randomized clinical trials is generalizable to the marketplace. Finasteride rates are inversely related to surgery rates, and tamsulosin rates are positively associated with surgery rates. Surgery rates are not significantly associated with urologists per capita. PMID- 16469611 TI - Consequences of iliac arterial atheroma on renal transplantation. AB - PURPOSE: Because recipient age has significantly increased in the last 15 years, surgeons must sometimes deal with atherosclerotic lesions of the iliac arterial system. Arterial restoration during renal transplantation should now be less frequent due to better preoperative screening and the prevention of arteriosclerosis in patients on renal transplantation waiting lists but in some patients EIA atheroma may require an additional surgical vascular procedure during renal transplantation. We describe the role of iliac artery atherosclerosis and the technical aspects of arterial restoration performed in patients who have undergone renal transplantation since 1985. MATERIALS AND METHODS: In a series of 1,110 cadaveric renal transplantations performed between 1985 and 2000, 38 patients required endarterectomy during renal transplantation and 69 were considered not to require any special procedure. RESULTS: In the 38 patients requiring endarterectomy a total of 12 end-to-end arterial anastomoses were performed and 6 ASs (50%) were observed, while 26 side-to-end arterial anastomoses were performed with only 1 AS (4%). Patient and graft survival curves showed a significant negative correlation with the severity of atherosclerosis. CONCLUSIONS: Preoperative assessment of the EIA is mandatory before renal transplantation. Renal transplantation can be performed in patients with an atheromatous EIA if the artery can be clamped for endarterectomy. In our experience side-to-end anastomosis using a donor patch onto the EIA provides better results by avoiding AS after endarterectomy. However, despite vascular repair graft survival is significantly lower in patients with atheromatous lesions requiring endarterectomy. PMID- 16469612 TI - Penile prosthetic surgery in neurologically impaired patients: long-term followup. AB - PURPOSE: Penile prosthetics are a viable option for erectile dysfunction in neurologically impaired patients. Penile implants can also be used to facilitate the management of urinary drainage when penile retraction has made this difficult. MATERIALS AND METHODS: Between 1980 and 1996, 245 neurologically impaired patients with a mean age of 40.8 years (range 16 to 75), including 188 with paraplegia, 57 with quadriplegia and 197 with spinal cord injuries, were treated for erectile dysfunction and/or urinary incontinence with penile prosthesis implantation. The mean history of paralysis was 11.2 years (range 1 to 52). After neuro-urological evaluation all patients included in this study were considered candidates for penile prosthesis implantation. A followup program for treatment success, patient satisfaction, problems and complications was subsequently initiated. RESULTS: During 17 years a total of 293 surgical procedures in 245 patients were done with the implantation of 147 semirigid (Jonas), 113 self-contained inflatable (Dynaflex) and 33 inflatable 3-piece (AMS 700) prostheses. There were 3 patient groups based on the indication for penile prosthetic surgery, namely group 1-134 patients with urinary management only, group 2-60 with erectile dysfunction only, and group 3-51 with urinary management and erectile dysfunction. At a mean followup of 7.2 years (maximum 17) 195 patients were reevaluated in clinic. In 122 patients (90.3%) urinary management problems were resolved. Erectile dysfunction treatment was successful in 76 patients (82.6%). There were 43 revisions for technical reasons and infections. The infection rate was 5% (12 patients). The perforation rate was different for different implant devices, that is 18.1% (15 of 83 cases) for semirigid devices, 2.4% (2 of 84) for self-contained inflatable devices and 0% (0 of 28) for inflatable 3-piece devices. CONCLUSIONS: The implantation of a penile prosthesis is a safe procedure for erectile dysfunction and/or urinary incontinence in neurologically impaired patients. Based on technical advances the complication rates significantly decreased during the years. The implantation of an inflatable 3-piece penile prosthesis in a neurologically impaired patient is a safe and viable procedure. Indications include the management of erectile dysfunction and problematic urinary collection. PMID- 16469614 TI - Increased expression of hypoxia-inducible factor-1alpha in the internal spermatic vein of patients with varicocele. AB - PURPOSE: Varicocele is recognized as a cause of male infertility. Testis hypoxia may be one of the possible mechanisms of varicocele. We examined whether tissue hypoxia occurred in the ISV of patients with varicocele by detecting the expression of HIF-1alpha. MATERIALS AND METHODS: The study group consisted of 8 patients with grade 3 left varicocele. The control group consisted of 6 volunteers with left indirect inguinal hernia. Using a left inguinal surgical incision, a 1 cm section of ISV was resected from each patient in both groups as specimens for immunoblotting and immunohistochemical staining of HIF-1alpha. Results were analyzed using Student's t test. RESULTS: HIF-1alpha immunoblots from both groups revealed a single band. The relative intensity of the HIF-1alpha protein band was 10.92 +/- 2.70 in the control group and 73.15 +/- 8.93 in patients with varicocele (ie 7-fold higher). CONCLUSIONS: HIF-1alpha expression in the ISV of patients with varicocele was significantly higher than in the control group. This directly shows that hypoxia related pathophysiological changes have occurred in the ISV of patients with varicocele and that hypoxia may have also occurred in the testicular tissue. Thus, it would be of interest to investigate whether decreasing HIF-1alpha activation and testis hypoxia could reduce the recurrence of varicocele. To our knowledge, this is the first report on HIF-1alpha expression in human ISV. Additional studies will be necessary to clarify the relationship between testis hypoxia and male infertility in patients with varicocele. PMID- 16469616 TI - Optimizing the outcome of microsurgical subinguinal varicocelectomy using isosulfan blue: a prospective randomized trial. AB - PURPOSE: Varicoceles are the main cause of correctible male factor infertility. Loss of testicular volume and abnormal seminal parameters are indications for varicocelectomy. We assessed a prospective randomized trial comparing standard microsurgical varicocele repair with the microsurgical approach using isosulfan blue. We determined if the use of isosulfan blue reduces the incidence of postoperative hydroceles following microsurgical varicocele repair. MATERIALS AND METHODS: Two groups of 25 consecutive patients each were randomized. Indications for surgery included fertility problems as well as scrotal pain and difference in testicular size. All patients underwent microsurgical repair using the subinguinal approach. Patients in group 1 underwent standard microsurgery. Those in group 2 had additional isosulfan blue administration. All were investigated 3 and 6 months postoperatively regarding varicocele recurrence, hydrocele, atrophy, pain or other complications. RESULTS: There were no intraoperative complications. At 3 months postoperatively the incidence of hydrocele in group 1 was 20% decreasing to 16% at 6 months, whereas in group 2 no hydrocele was detectable. There were 2 patients in each group who reported ongoing pain, and 1 in each presented with recurrent varicocele. No atrophy was seen. In 1 patient blue pigmentation of the left hemiscrotum was seen at 3 months but vanished at 6 months. CONCLUSIONS: Microsurgical repair of varicoceles using isosulfan blue helps in identifying and preserving the lymphatic drainage. It prevents postoperative hydrocele, the most common postoperative complication, without any supplementary risk to the patient. Additionally, testicular edema causing impaired spermatogenesis can be avoided. PMID- 16469618 TI - The International Consultation on Incontinence Modular Questionnaire: www.iciq.net. AB - PURPOSE: In 1998 the first ICI was held in Monaco, sponsored by WHO and organized by the International Continence Society and International Consultation on Urological Diseases. The Scientific Committee recognized the need to develop a universally applicable questionnaire for wide application across international populations in clinical practice and research to assess urinary incontinence, facilitating the comparison of findings from different settings and studies, in a manner similar to the International Prostate Symptom Score. MATERIALS AND METHODS: An Advisory Board was formed to steer the development of the ICIQ and a decision was made to extend the concept further, developing the ICIQ Modular Questionnaire. RESULTS: The first module developed was the ICIQ Short Form Questionnaire for urinary incontinence. ICIQ modules have been developed or adapted for urinary tract symptoms and they are being developed for vaginal and lower bowel symptoms. Additional sexual matters and quality of life modules will become available for each condition area. Modules to assess patient satisfaction are expected to be of particular use for assessing treatment effectiveness. The ICIQ Advisory Board recently proposed the development of the ICIQ website, which is anticipated to be crucial for informing potential users of the phase of development of all ICIQ modules. CONCLUSION: The ICIQ can offer a full range of urinary tract symptom questionnaires. The website will aim to attract collaborators committed to the concept of this internationally accepted modular questionnaire who are willing to help with its development. PMID- 16469617 TI - Self-esteem, confidence and relationship satisfaction of men with erectile dysfunction treated with sildenafil citrate: a multicenter, randomized, parallel group, double-blind, placebo controlled study in the United States. AB - PURPOSE: We assessed the change in confidence, relationships and self-esteem, and its correlation with erectile function in men with ED treated with sildenafil citrate in the first United States based, double-blind, placebo controlled, randomized trial assessed by the validated SEAR. MATERIALS AND METHODS: This 12 week flexible dose (25, 50 or 100 mg) trial determined change scores from baseline to end of treatment for the 5 SEAR components (Sexual Relationship domain, Confidence domain, Self-Esteem subscale [prespecified as the primary end point], Overall Relationship subscale and Overall score), and their correlations with the IIEF and event log data, as well as correlations between SEAR components and a general efficacy question at the end of treatment. RESULTS: Compared with the placebo group (125 patients, mean age +/- SD 55 +/- 13 years, mean years ED 3.8 +/- 4.2), the sildenafil group (128 patients, mean age +/- SD 56 +/- 12, mean years ED 4.6 +/- 4.3) had significantly greater improvements in all 5 SEAR components (p < 0.0001) and all sexual function measures. SEAR component scores showed significant correlations with IIEF Erectile Function domain scores (r range 0.34 to 0.69, p < 0.0001), other IIEF domain scores (p < 0.0001), percentage of successful intercourse attempts (p < 0.0001) and frequency of erection that allowed satisfactory intercourse (p < 0.0001). CONCLUSIONS: In this study of men with ED, sildenafil produced substantial improvements in self esteem, confidence and relationship satisfaction as measured by SEAR scores, which showed moderate to high positive correlations with IIEF scores. PMID- 16469620 TI - Predictors of medication adherence and associated health care costs in an older population with overactive bladder syndrome: a longitudinal cohort study. AB - PURPOSE: We examined the relationship between self-reported health status data, subsequent antimuscarinic medication adherence and health care service use in older adults with OAB syndrome in a managed care setting. MATERIALS AND METHODS: This was a longitudinal cohort study of older adults in the southeastern United States with OAB who completed a health status assessment, used antimuscarinic medications and were enrolled in an HMO continuously for 1 to 3 years. Demographic, clinical and use related economic variables were also retrieved from the administrative claims data of patient HMOs. Prescription refill patterns were used to measure medication adherence. Associations were examined with a sequential, mixed model regression approach. RESULTS: A total of 275 patients were included. The severity of comorbidity (Charlson index), patient perception of quality of life (Short Form-12 scores) and total number of prescribed medications during the year prior to enrollment in a Medicare HMO were independently associated with decreased antimuscarinic MPRs after enrollment. After controlling for other variables increased antimuscarinic MPR remained the strongest predictor of decreased total annual health care costs (5.6% decrease in annual costs with every 10% increase in MPR, p < 0.001). CONCLUSIONS: We found strong associations between decreased antimuscarinic medication adherence and increased health care service use in older adults with OAB in a managed care setting. Health status assessments completed at enrollment had the potential to identify enrollees at higher risk for nonadherent behaviors and poor health related outcomes. PMID- 16469622 TI - Hydrocele formation following laparoscopic varicocelectomy. AB - PURPOSE: Hydrocele is a known complication of varicocelectomy. We evaluated the incidence of hydrocele following laparoscopic varicocelectomy at our institution. MATERIALS AND METHODS: A total of 89 boys were treated with laparoscopic ligation of the spermatic vessels for clinically palpable varicoceles between January 2000 and December 2003. Charts were retrospectively reviewed. A total of 10 patients were excluded because they were lost to followup or presented with recurrent varicocele. Followup consisted of office visits with physical examinations at 1 and 12 months postoperatively. Patient charts were reviewed for perioperative variables, operative technique and complications. RESULTS: Only 1 of 79 patients (1.3%) had persistent varicocele with a mean of 20.7 months of followup. A total of 18 patients (22.8%) had development of hydrocele postoperatively, of whom 9 required hydrocelectomy. In addition, 2 of these 9 patients needed repeat hydrocelectomy. Of the 57 patients with greater than 6 months of followup 29.8% had development of hydrocele. A higher rate of hydrocele formation (31.1%) was also noted in patients who underwent ligation and division of the spermatic vessels rather than ligation alone (11.8%, p = 0.04). CONCLUSIONS: Our series demonstrates a high rate of hydrocele formation following laparoscopic varicocelectomy, particularly in patients with longer followup. The incidence of hydrocele after laparoscopic varicocelectomy may be underreported. However, there appears to be a statistically significant decrease in hydroceles when the internal spermatic vessels are simply ligated rather than ligated and divided. Despite its ease and low failure rate, the standard technique of laparoscopic varicocelectomy requires reexamination, potentially allowing modifications that may decrease hydrocele formation, such as salvaging lymphatics and avoiding division of the vessels. PMID- 16469623 TI - Ureteral access sheath for the management of pediatric renal and ureteral stones: a single center experience. AB - PURPOSE: Ureteral access sheaths were developed to facilitate difficult ureteroscopic access in adults. We report our use of the ureteral access sheath during standard ureteroscopic procedures in the pediatric stone population. MATERIALS AND METHODS: Eight patients underwent flexible ureteroscopy and holmium laser lithotripsy for symptomatic renal and ureteral calculi between October 2003 and October 2004. Average patient age was 9.3 years (range 4 to 13). There were 4 males and 4 females. Five of 8 patients had bilateral stones and underwent bilateral endoscopic stone treatment at the same operation. A 14Fr 20 cm Flexor ureteral access sheath was used for dilation and access of the ureter. Flexible pediatric ureteroscope was used to enter the ureter and collecting system for stone retrieval and fragmentation. RESULTS: All patients were rendered stone-free in a single operative setting. Average operative time was 99 minutes (range 65 to 130). Average total stone burden for the group was 9 +/- 3 mm. All patients received stents after the procedure. There were no perioperative complications. All patients were followed with renal-bladder ultrasound and plain x-ray. All patients were stone-free at a mean followup of 10 months (median 11). CONCLUSIONS: Routine use of a ureteral access sheath in children facilitates flexible ureteroscopy with lithotripsy of multiple renal and proximal ureteral stones. The procedure is safe and time efficient, and is associated with minimal morbidity. Larger prospective studies in children with longer followup are warranted. PMID- 16469625 TI - Correction of hypospadias with a vertical preputial island flap: a 23-year experience. AB - PURPOSE: We report our experience using the preputial island flap technique (Scuderi procedure) to correct penile hypospadias. MATERIALS AND METHODS: A total of 152 patients underwent repair between 1982 and 2004. Nine patients (6%) had proximal hypospadias, 46 (30%) had mid penile hypospadias and 97 (64%) had distal hypospadias. A total of 146 patients (96%) had not previously undergone surgical treatment, while 6 (4%) had undergone surgery. RESULTS: After the primary repair 3 patients had fistula and 10 had mild stenosis. The immediate success rate was 91.4% (139 of 152 patients), which later increased to 98% (149 of 152) after nonsurgical treatment of the stenoses. CONCLUSIONS: Preputial island flap urethroplasty is a versatile operation that corrects hypospadias and is particularly indicated if there is associated severe penile curvature, with a low complication rate and superior cosmetic results. PMID- 16469627 TI - Simplified technique to create a concealed catheterizable stoma: the VR flap. AB - PURPOSE: We describe a surgical modification of VQZ plasty, the VR flap. In addition to avoiding skin surface mucosal prolapse and preventing stenosis, this procedure is easier to describe and perform. MATERIALS AND METHODS: A total of 12 patients (4 males and 8 females) 6 to 14 years old (mean age 11.6) underwent simultaneous creation of a continent catheterizable channel using the Mitrofanoff principle and MACE procedure. VQZ plasty was modified by using a quadrilateral flap positioned at a 90-degree angle from the V flap instead of immediately next to it. The quadrilateral flap is used to create a skin channel allowing the use of a shorter conduit and avoiding exposed mucosa. The modification creates a linear closure. RESULTS: A total of 24 CCCs (Mitrofanoff 12, MACE 12) were done simultaneously in 12 patients. Of the channels 12 were implanted in the umbilicus (CCC 6, MACE 6) and 12 in a stoma created by the VR flap. There were no significant complications. One patient required surgical revision and 1 required stomal dilation for stomal stenosis. All patients were continent through all channels. Mean followup was 18 months. CONCLUSIONS: This modification offers good cosmetic and functional results, and the outcome compares favorably with other series. The procedure is easy to understand and perform, and its application is not limited by the thickness of the abdominal wall. Moreover, it allows for an easier closure and limits the Z-shaped appearance of the closure. PMID- 16469628 TI - Orthotopic continent urinary diversion after radical cystectomy in pediatric patients with genitourinary rhabdomyosarcoma. AB - PURPOSE: We present our experience with orthotopic continent urinary diversion following radical cystectomy due to genitourinary rhabdomyosarcoma in children not responding to radiotherapy or chemotherapy. MATERIALS AND METHODS: Four children with persistent genitourinary rhabdomyosarcoma underwent radical cystectomy with reconstruction of an orthotopic ileal neobladder. RESULTS: Average followup was 50.7 months. One patient died at 12 months postoperatively due to pulmonary recurrence. All patients were able to void completely at regular intervals. Renal function was normal in all patients. Average reservoir capacity was 250 ml. Daytime and nighttime urinary continence was achieved in all patients. CONCLUSIONS: The orthotopic continent urinary diversion is technically feasible even in small children. It represents a safe long-term option and offers good quality of life after disfiguring surgery. In fact, patients are able to void spontaneously through the urethra. PMID- 16469630 TI - New contralateral vesicoureteral reflux following dextranomer/hyaluronic Acid implantation: incidence and identification of a high risk group. AB - PURPOSE: To our knowledge the incidence of NCVUR following the endoscopic treatment of VUR with Dx/HA has not been reported previously. We evaluated the outcomes in a group of patients to determine the incidence, and to attempt to identify risk factors. MATERIALS AND METHODS: A total of 126 children with primary unilateral VUR underwent unilateral Dx/HA implantation at our institutions. The incidence of NCVUR was determined by postoperative VCUG. Indications for surgery, patient age and gender, preoperative grade of VUR and volume of Dx/HA injected were assessed as possible risk factors for NCVUR. RESULTS: Of the patients 96 (76.2%) were female, and mean age was 4.8 years. The principal indications for Dx/HA implantation were persistent reflux in 56 patients (44.4%) and primary therapy in 51 (40.5%). At followup VCUG 17 patients (13.5%) had NCVUR. No variable independently appeared to influence the incidence of NCVUR. Statistical analysis suggests that females younger than 5 years have an increased incidence of NCVUR (13 of 62, or 21% vs 4 of 64, or 6.3% of the remaining patients, p = 0.016). CONCLUSIONS: NCVUR occurred in approximately 13% of our patients. Patients with higher preoperative VUR grade or a lower number of preoperative VCUGs and those undergoing treatment as primary therapy did not have an increased incidence. Girls younger than 5 years had the highest incidence of NCVUR, and initial bilateral injection may be a consideration for this group. Further effort directed at identifying the etiology and risk factors for NCVUR is needed. PMID- 16469632 TI - Repeated intradetrusor botulinum toxin type A in children with neurogenic bladder due to myelomeningocele. AB - PURPOSE: We evaluated the effect of repeated intradetrusor injections of BTA in pediatric myelomeningocele not responding to medical management. MATERIALS AND METHODS: After baseline history, physical examination and urodynamic assessment BTA was injected (5 IU/kg body weight, maximum 300 IU) at 10 to 30 sites. Clinical and urodynamic assessment was performed at 3 months after each injection. Re-treatment was offered when clinical symptoms returned. RESULTS: A total of 20 patients (average age 13 years) received BTA injections. Of the patients 13 became continent. MBC increased from 215.6 +/- 58.8 cc to 338.3 +/- 98.4 cc (p < 0.01), MDP decreased from 43 +/- 13.7 cm H2O to 21.6 +/- 10.5 cm H2O (p < 0.01) and compliance increased from 5.2 +/- 2.6 ml/cm H2O to 13 +/- 6.9 ml/cm H2O (p < 0.01). At an average of 8.1 months after the first injection all 13 patients received a second injection, which led to similar improvement, ie MBC increased from 200.5 +/- 41.6 cc to 404.2 +/- 57.8 cc (p < 0.001), MDP decreased from 48.18 +/- 6.1 cm H2O to 27.8 +/- 3.7 cm H2O (p < 0.01) and compliance increased from 6.0 +/- 3.1 ml/cm H2O to 15.1 +/- 5.2 ml/cm H2O (p < 0.01). Among the responders 3 received 3 injections and 1 received 4 injections, all of whom exhibited improvement similar to that seen initially. Among our initial cohort of 20 patients 7 failed to improve initially and 6 failed to improve after a second injection. CONCLUSIONS: BTA seems to be a simple and safe way to postpone or avoid invasive procedures in two thirds of children with myelomeningocele not responding to usual medical treatment. PMID- 16469633 TI - Endoscopic injection with polydimethylsiloxane for the treatment of pediatric urinary incontinence in the neurogenic bladder: long-term results. AB - PURPOSE: We report the long-term results of endoscopic injection of PDMS in correcting urinary incontinence in children with neurogenic bladder. MATERIALS AND METHODS: We performed a single center, retrospective study of patients receiving endoscopic injections of PDMS. All procedures were performed as outpatient surgery using endoscopic guidance. Procedure tolerance was assessed at 15 days, and efficacy was evaluated at 3 months, 6 months and annually thereafter. Success was defined as periods of dryness between bladder voiding of more than 4 hours during the daytime without the need to wear pads. Improvement was defined as periods of dryness greater than 2 and less than 4 hours with occasional protection. RESULTS: A total of 49 children (21 boys and 28 girls) have received 1 or more injections of PDMS since 1995. Etiology was spina bifida in 41 patients (84%), with surgery (enterocystoplasty and/or bladder neck reconstruction) performed previously in 27 patients. Mean patient age was 14 years (standard deviation 4.8). Mean volume of PDMS per treatment was 3.6 ml. At the end of the 6-year mean followup 16 patients (33%) were continent and 7 (14%) were improved. Continence was unchanged in the remaining 26 patients (53%). After a significant deterioration of the results the outcome remained almost unchanged from 18 months of followup. Bladder neck surgery, bladder hyperactivity if medically controlled and gender have no influence statistically on the long-term results. CONCLUSIONS: Injection of PDMS for incontinence of neurogenic origin is a reliable technique that achieves long-term continence in almost a third of the patients. This procedure can be used either as first line treatment or in addition to other surgical techniques. The initial success of the procedure seems predictive of success in the long term, and results are stable after 18 months of followup. PMID- 16469634 TI - Cost analysis of neonatal circumcision in a large health maintenance organization. AB - PURPOSE: We studied the costs of newborn circumcision in relation to its health benefits later in life. MATERIALS AND METHODS: We conducted a retrospective database analysis using direct internal cost data from Kaiser Permanente Northern California-a large health maintenance organization-and published cost data (including the cost of medically indicated postneonatal circumcision). The study cohort consisted of 14,893 male infants born in 1996. One-way sensitivity analysis was used to demonstrate the impact of selected variables in the model. Monte Carlo analysis was used to determine the 95% confidence intervals. RESULTS: Postneonatal circumcision was 10 times as expensive as neonatal circumcision (1,921 dollars per infant vs 165 dollars per newborn), and was medically indicated for 9.6% of uncircumcised males. Cost benefits of circumcision resulted from prevention of infant urinary tract infection, balanoposthitis, phimosis, HIV infection and penile cancer. Assuming initial neonatal circumcision cost to be 200 dollars, the future health care cost offset (avoided) was calculated as 183 dollars (range 93 dollars to 303 dollars in 95% of simulations). CONCLUSIONS: Multiple lifetime medical benefits of neonatal circumcision can be achieved at little or no cost. Because postneonatal circumcision is so expensive, its rate is the most important factor determining future cost savings from newborn circumcision. PMID- 16469635 TI - Intestinal metaplasia of the bladder. PMID- 16469636 TI - Nutcracker phenomenon. PMID- 16469637 TI - Bipolar and multipolar radio frequency ablation with resistance controlled power output: standardized ex vivo kidney tissue evaluation. AB - PURPOSE: We investigated a newly developed bipolar and multipolar RF ablation system with an internally cooled electrode and resistance controlled power output in a standardized model of perfused ex vivo kidney tissue. MATERIALS AND METHODS: RF energy was applied at different power levels (20, 30 and 60 W) for 1, 3, 5 and 9 minutes. Each treatment parameter was repeated 5 times. For the 20/30 W levels a bipolar electrode with an active conducting part of 20/30 mm was selected. At 60 W 2 bipolar electrodes with an active conducting part (30 mm each) were connected. Lesion volumes and shapes were calculated by measuring the maximum vertical, long axis and short axis diameters of the macroscopic lesion. RESULTS: Lesion volume increased significantly with the treatment time and generator power applied (p < 0.0001). Lesion size in multipolar ablated zones was larger than that in bipolar ablated zones. A reliable dose-effect relationship existed between the generator power/applied treatment time and ablated tissue lesion size. All lesions were elliptical. CONCLUSIONS: Bipolar and multipolar RF ablation with an internally cooled electrode and tissue resistance control represent an interesting advance in RF technology. The development of lesion size and volume is predictable, while a uniform lesion shape can be achieved in perfused ex vivo kidney tissue. Further in vivo trials are required to test whether complete and reliable tumor tissue ablation is possible with this system. PMID- 16469638 TI - Gene expression profiling of ERBB receptors and ligands in human transitional cell carcinoma of the bladder. AB - PURPOSE: The ErbB driven growth pathway has been implicated in most human epithelial malignancies. Therefore, its blockade is a promising therapeutic strategy and several candidate drugs are currently undergoing clinical trials. Paradoxically little is known of the expression pattern or clinical significance of the 4 ErbB receptors and their 11 ligands in TCC of the bladder. MATERIALS AND METHODS: To obtain further insight into the molecular pathogenesis of TCC we used quantitative real-time reverse transcriptase-polymerase chain reaction assay to quantify mRNA expression of the 4 ERBB and their 11 known ligand genes, including recently described EPGN/epigen, in 73 tumor samples. RESULTS: The level of mRNA of 4 ligand genes (EGF, NRG1, NRG2 and NRG3) was extremely low, that is detectable but not quantifiable. Six genes were over expressed (ERBB2, TGFA, HB EGF, AREG, EREG and EPGN), 3 were under expressed (ERBB1, ERBB4 and NRG4) and 2 were over or under expressed (ERBB3 and BTC). ERBB2 and AREG expression differed between early stage tumors (pTa grade 1) and normal samples. The most marked differences in expression were ERBB3, EREG and NRG4 between superficial and muscle invasive tumors (p = 0.0069, 0.00007 and 0.0000001, respectively), and TGFA and NRG4 between low and high grade superficial tumors, and between pT1 or greater and pTa tumors. CONCLUSIONS: This study shows the involvement of the ERBB family and ligand genes in TCC. Most receptor and ligand genes are deregulated at different stages of carcinogenesis, implying that they should be studied simultaneously. Quantitative real-time reverse transcriptase-polymerase chain reaction could be used to determine ErbB signaling pathway status in individuals with a view to tailored therapy. PMID- 16469639 TI - Characterization of a panel of cell lines derived from urothelial neoplasms: genetic alterations, growth in vivo and the relationship of adenoviral mediated gene transfer to coxsackie adenovirus receptor expression. AB - PURPOSE: Cell lines have become an essential component for the investigation of cancer. We have developed a panel of cell lines derived from human urothelial cancers and we describe some of their important characteristics. MATERIALS AND METHODS: Ten human urothelial cancer cell lines were characterized by their growth in athymic nude mice, CAR expression and their susceptibility to adenoviral mediated transfer of the green fluorescence protein gene. TP53 mutation status and immunochemical analysis of p53, pRB and p16 were also examined. RESULTS: Five cell lines rapidly produced tumors in athymic nude mice. Two cell lines produced tumors in 1 month, 1 produced them in 3 months and 2 were nontumorigenic. The cell lines varied in CAR expression and in their susceptibility to adenoviral mediated gene transduction. There was no direct correlation between CAR expression and susceptibility to adenoviral mediated gene transduction. Seven cell lines had TP53 mutations, of which 2 had large deletions and did not express p53 protein by immunostaining. All cell lines expressed abnormal pRB by immunochemical analysis (3 had no staining and 7 had homogenously strong staining) and 8 did not express p16 (7 showed homogeneously strong pRB staining). CONCLUSIONS: Our panel of 10 human urothelial cell lines differed in genetic alterations, growth in nude mice, susceptibility to adenoviral mediated gene transduction, and expression of p53, p16 and pRB. The availability of various urothelial cancer cell lines with differing genotypic and phenotypic features will facilitate further research into bladder cancer. PMID- 16469640 TI - Botulinum toxin type a inhibits calcitonin gene-related peptide release from isolated rat bladder. AB - PURPOSE: Increasing evidence suggests that sensory nerve dysfunction may underlie several urological disorders, including interstitial cystitis and sensory urgency. We determined the effect of botulinum toxin type A (Allergan, Irvine, California) on baseline and chemically evoked release of the sensory neuropeptide, calcitonin gene-related peptide in an isolated bladder preparation. MATERIALS AND METHODS: Whole rat bladders were incubated in a series of tissue baths containing physiological salt solution. Following bladder equilibration in PSS sequential incubation was performed and this sample was used to measure baseline CGRP release. To evoke CGRP release tissue was subsequently incubated in PSS containing capsaicin (30 nM) and adenosine triphosphate (10 microM). To measure the effect of BTX-A on baseline and evoked CGRP release bladders were incubated for 6 hours in an organ bath containing BTX-A (50 microM) or vehicle prior to bladder equilibration. CGRP release was determined by radioimmunoassay. RESULTS: Mean baseline release of CGRP +/- SEM was 346 +/- 44 pg/gm. Adenosine triphosphate/capsaicin application increased CGRP release by 75% over baseline (606 +/- 98 pg/gm, p < 0.005). BTX-A application resulted in a 19% decrease in baseline release of CGRP, although this difference did not achieve statistical significance. BTX-A application significantly decreased evoked CGRP by 62% vs control (606 +/- 98 vs 229 +/- 21 pg/gm, p < 0.005). CONCLUSIONS: BTX-A application inhibits the evoked release of CGRP from afferent nerve terminals in isolated rat bladder. This finding suggests a potential clinical benefit of BTX-A for the treatment of interstitial cystitis or sensory urgency. PMID- 16469641 TI - Molecular size affects urine excretion of pentosan polysulfate. AB - PURPOSE: In human subjects only a small percent of oral PPS is found in urine. Commercially available PPS is a heterogeneous mixture with varying molecular weights. Our hypothesis was that only the low molecular weight fraction reaches the urine. MATERIALS AND METHODS: Urine was obtained from patients with IC who were chronically receiving PPS. The amount and molecular size of PPS in the urine were determined by enzyme-linked immunosorbent assay and molecular sieve chromatography. PPS was purified from Elmiron capsules and fractionated into LMW and HMW fractions. Urine recovery of PPS was measured in rabbits after oral or intravenous administration of unfractionated, LMW or HMW PPS. RESULTS: The median urine PPS level in 34 patients with IC was 1.2 microg/ml (range 0.5 to 27.7). All PPS recovered from IC urine was LMW. After intravenous administration in rabbits the median recovery in urine was 47.2% (range 19.7% to 73.2%) for unfractionated PPS, 74.6% (range 31.4% to 96.3%) for LMW and 3.3% (range 2.5% to 5.0%) for HMW. After oral administration in rabbits the median recovery in urine was 7.4% (range 2.1% to 46.0%) for LMW and 0.10% (range 0.0% to 0.3%) for HMW. CONCLUSIONS: In patients with IC who are on oral PPS the PPS recovered in the urine is all of LMW. In rabbits the HMW fraction of PPS is recovered in small amounts from urine after intravenous administration and not at all after oral administration. PMID- 16469642 TI - Maternal separation uncouples reflex from spontaneous voiding in rat pups. AB - PURPOSE: Rat pups only void when the perigenital-bladder reflex is activated by the mother rat licking the perineum. Maternal separation causes bladder distention as well as stress responses and anxiety behaviors in adult rats. We determined if MS would change voiding reflex maturation in neonatal rats. MATERIALS AND METHODS: A total of 14 Sprague-Dawley rat pups were subjected to 6 hours of daily MS and 17 were subjected to 6 hours of MS with bladder emptying by perigenital stimulation at 3 hours on postnatal days 2 to 14. Age matched controls for the 2 groups remained with the mother. Spontaneous voiding in awake pups from 1 to 3 weeks was monitored in a metabolic cage and perigenital-bladder reflex latency was determined from 1 to 7 weeks. Cystometry was performed at 9 weeks with the rats under urethane anesthesia. RESULTS: Spontaneous voiding began at 3 weeks in all animals. The latency of the perigenital-bladder reflex at 3 weeks was shorter than the latency at 2 days in MS animals (3.3 vs 6.4 seconds, p < 0.01) but not in control or MSPG animals. MS animals maintained the perigenital bladder reflex 2 weeks longer than control animals. The spontaneous voiding behavior of MSPG animals was similar to that in controls. CONCLUSIONS: Intermittent bladder distention delays withdrawal of the spinal perigenital bladder reflex but it does not affect maturation of the supraspinal bladder bladder reflex that controls spontaneous voiding in older rats. This suggests that increased bladder afferent firing can selectively modulate spinal but not supraspinal mechanisms controlling postnatal changes in voiding function. PMID- 16469643 TI - Expression of smoothelin in the normal and the overactive human bladder. AB - PURPOSE: We established the expression pattern of smoothelin, a marker protein for contractile smooth muscle cells, in the human detrusor and investigated its possible impact on bladder overactivity. MATERIALS AND METHODS: Detrusor samples of 13 overactive bladders (sensory urge and detrusor instability) were obtained before botulinum toxin injection and compared to those of 8 normally contractile, nonobstructed bladders obtained during radical cystectomy. Smoothelin mRNA expression patterns were investigated by Northern blot and variant specific reverse transcriptase-polymerase chain reaction as well as by quantitative reverse transcriptase-polymerase chain reaction on laser capture, microdissected smooth muscle. At the protein level smoothelin was investigated by standard and quantitative immunohistochemistry. RESULTS: The bladder muscularis expressed vascular and visceral smoothelin isoforms, and 2 of the known splice variants. In the smooth muscle of patients with detrusor instability and sensory urge a significant 2.4 and 2.2-fold increase, respectively, in smoothelin variant 1 mRNA was observed in comparison to that of normal controls. Analyses at the smoothelin protein level confirmed significant up-regulation in these bladder dysfunctions by a factor of 2.3 and 1.8, respectively. No significant difference in smoothelin expression was observed between detrusor instability and sensory urge. CONCLUSIONS: Increased expression of smoothelin in patients with detrusor instability and sensory urge implies that the etiology of these dysfunctions includes changes in myogenic parameters. In addition, our data support the new classification of the International Continence Society for overactive bladder proposing that sensory urge and detrusor instability represent a single clinical entity. PMID- 16469644 TI - Novel action of botulinum toxin on the stromal and epithelial components of the prostate gland. AB - PURPOSE: Intraprostatic injection of BTX-A has demonstrated clinical improvement in men with bladder outlet obstruction. We investigated the mechanisms of action of BTX-A on the prostate. MATERIALS AND METHODS: Adult male Sprague-Dawley rats were injected with varying doses of BTX-A into the prostate and the prostates were harvested after 1 or 2 weeks. The effects of BTX-A on prostate histology, and the proliferative and apoptotic indexes were determined using hematoxylin and eosin staining, proliferative cell nuclear antigen staining and TUNEL staining, respectively. Changes in alpha(1A) adrenergic receptor and androgen receptor were evaluated by Western blotting. RESULTS: One week after BTX-A injection generalized prostate atrophy was observed. There was a significant increase in apoptotic cells (12, 16 and 22-fold), and decrease in proliferative cells (38%, 77% and 80%) and alpha(1A) adrenergic receptor (13%, 80% and 81%) for 5, 10 and 20 U, respectively. There was no significant change in androgen receptors. The effects were decreased 2 weeks after BTX-A treatment. CONCLUSIONS: BTX-A injection into the prostate alters cellular dynamics by inducing apoptosis, inhibiting proliferation and down-regulating alpha(1A) adrenergic receptors. BTX A may potentially be the drug that has dual actions on the static and dynamic components of benign prostatic hyperplasia. PMID- 16469645 TI - Role of nitric oxide in the relaxation elicited by sildenafil in penile resistance arteries. AB - PURPOSE: We investigated the role of the vascular endothelium and the L arginine/nitric oxide pathway in the vasorelaxant effect of the phosphodiesterase type 5 inhibitor sildenafil in penile resistance arteries. MATERIALS AND METHODS: Second or third order branches of the horse deep intracavernous penile artery were mounted in microvascular myographs. The vasodilator effects of sildenafil and the NO donor SNAP (S-nitrosoacetyl-D,L-penicillamine) were evaluated in the absence and presence of the endothelium and inhibitors of the NO/cGMP (cyclic guanosine 3',5'-monophosphate) pathway. RESULTS: In phenylephrine precontracted, endothelium intact arteries sildenafil elicited potent relaxations that were markedly decreased by the blockade of soluble guanylate cyclase with ODQ (1H [1,2,4]oxadiazolo[4-3a]quinoxalin-1-one). Endothelium removal and the inhibition of NO synthase with N(G)-nitro-L-arginine (L-NOARG) caused pronounced inhibition of sildenafil elicited relaxations but not of SNAP induced responses. Combined treatment with the cyclooxygenase blocker indomethacin and L-NOARG caused significantly greater inhibition of sildenafil relaxations than that produced by L-NOARG alone. Inhibitors of the cGMP (PKG) and the cAMP (cyclic adenosine 3',5' monophosphate) dependent protein kinases Rp-8-Br-PET-cGMPS (beta-phenyl-1, N2 etheno-8-bromoguanosine-3',5'-cyclic monophosphorothioate, Rp-isomer) and Rp-8 CPT-cAMPS (Rp-8-CPT-cAMPS (8-(4-chlorophenylthio) adenosine-3',5'-cyclic monophosphorothioate, Rp-isomer), respectively, inhibited the sildenafil concentration-relaxation curves. The relaxant responses of SNAP were markedly decreased by PKG inhibitor and to a lesser extent by cAMP dependent protein kinase inhibitor. CONCLUSIONS: The current results demonstrate a potent relaxant effect of sildenafil in penile resistance arteries due in part to cGMP accumulation and to the enhanced effects of basal released, endothelial derived NO acting through PKG activation. Cross-activation of the cAMP signaling pathway by sildenafil is also suggested. PMID- 16469646 TI - Re: Complete primary repair of bladder exstrophy in children presenting late and those with failed initial closure: single center experience. PMID- 16469647 TI - Re: Transcutaneous electrocavernosography: a tool for recording the electromyographic activity of the corpora cavernosa. PMID- 16469648 TI - Re: Regional lymph node staging using lymphotropic nanoparticle enhanced magnetic resonance imaging with ferumoxtran-10 in patients with penile cancer. PMID- 16469649 TI - Re: Oxidative stress in arteriogenic erectile dysfunction: prophylactic role of antioxidants. PMID- 16469651 TI - Anatomy and physiology of the larynx. PMID- 16469652 TI - Diagnostic and therapeutic pitfalls in phonosurgery. PMID- 16469653 TI - Surgical management of sulcus vocalis and vocal fold scarring. PMID- 16469654 TI - Injection laryngoplasty. PMID- 16469655 TI - Practical applications of laryngeal framework surgery. PMID- 16469656 TI - Phonosurgery for pitch alteration: feminization and masculinization of the voice. PMID- 16469657 TI - Laryngeal dystonia. PMID- 16469658 TI - Treatment of adductor spasmodic dysphonia with selective laryngeal adductor denervation and reinnervation surgery. PMID- 16469659 TI - Office-based laryngeal procedures. PMID- 16469660 TI - Contemporary management of laryngeal papilloma in adults and children. PMID- 16469661 TI - Laser applications in laryngology: past, present, and future. PMID- 16469662 TI - Endoscopic treatment for early glottic cancer: indications and oncologic outcome. PMID- 16469663 TI - Reconstruction of glottic defects after endoscopic cordectomy: voice outcome. PMID- 16469664 TI - Eosinophilic esophagitis: its role in aerodigestive tract disorders. PMID- 16469666 TI - Clinical governance in gynaecological surgery. AB - The main elements of clinical governance are defining standards of high-quality care and then achieving high-quality care by reviewing practice in relation to these standards and effecting change as required to improve services. This chapter examines the current framework for clinical governance in the UK. The roles of the various national organisations involved in the implementation of clinical governance are described. The role of audit, risk management, appraisals and inspections in this process are discussed. The application of national standards of care to gynaecological surgery is then considered, with a focus on relevant guidelines and on the legal concepts of consent and confidentiality. PMID- 16469667 TI - Tuberculous enteritis presenting as small-bowel obstruction. PMID- 16469668 TI - Dysphagia as a clue to mucous membrane pemphigoid. PMID- 16469669 TI - Von Meyenburg complexes (bile duct hamartomas). PMID- 16469670 TI - Stability of human methanogenic flora over 35 years and a review of insights obtained from breath methane measurements. AB - BACKGROUND & AIMS: Breath methane measurements reflect the in situ activity of the methanogenic colonic flora. Thirty-five years ago we found that 34% of healthy adult subjects were methane producers (breath methane level >1 ppm above atmospheric methane levels). The current study presents a new survey of breath methane levels designed to determine if the activity of the methanogenic flora has changed over the past 35 years. In addition, we review insights into the methanogenic flora that have resulted from breath methane measurements. METHODS: The end-alveolar breath methane concentrations of 212 healthy adults living in the Minneapolis area were determined via gas chromatography. The influence of sex, age, and bowel movement frequency on methane production was assessed. RESULTS: The findings that 36.4% of participants were methane producers, with a mean methane concentration in these producers of 16.6 ppm, are strikingly similar to the values of 33.6% and 15.2 ppm observed 35 years ago. Neither sex nor age showed a statistically significant relationship to methane production. There was a negative correlation between frequency of bowel movements and breath methane concentration in methane producers. CONCLUSIONS: The activity of the methanogenic flora of healthy adults remained remarkably stable over the past 35 years despite widespread antibiotic use and dietary changes. A literature review revealed that many associations have been shown between methane production and clinical states, but it remains to be determined if methanogens actively influence human physiology or are simply a marker of colonic function. PMID- 16469671 TI - Effects of Helicobacter pylori and nonsteroidal anti-inflammatory drugs on peptic ulcer disease: a systematic review. AB - BACKGROUND & AIMS: The aim was to systematically review the interactions between Helicobacter pylori (HP) infection and NSAID use on the risk of uncomplicated or bleeding peptic ulcer. METHODS: All relevant full articles published in MEDLINE from January 1989-June 2004 were included. Sensitivity analyses for type of controls or use of aspirin or non-aspirin NSAIDs were performed. RESULTS: In 21 studies involving 10,146 patients, uncomplicated peptic ulcer was more common in HP-positive than HP-negative patients (pooled odds ratio [OR], 2.17) or in HP positive than HP-negative NSAID users (OR, 1.81). In 6 age-matched controlled studies, ulcer was more common in HP-positive than HP-negative patients (OR, 4.03), irrespective of NSAID use, and in NSAID users than non-users (OR, 3.10), irrespective of HP status; the risk of ulcer was 17.54-fold higher in HP-positive NSAID users than HP-negative non-users. The use of aspirin or non-aspirin NSAIDs did not affect the results. Ulcer bleeding was evaluated in 17 studies involving 4084 patients. NSAID use was more frequent in bleeding patients than control subjects (OR, 5.13), irrespective of HP status and type of controls. In contrast, HP infection in bleeding patients compared with control subjects was less frequent in the 8 studies with ulcer cases as control subjects (OR, 0.40) and more frequent in the 9 studies with uninvestigated subjects as controls (OR, 2.56). In the latter studies, presence compared with the absence of both HP and NSAIDs increased the risk of bleeding 20.83-fold. CONCLUSION: HP infection and NSAID use represent independent and synergistic risk factors for uncomplicated and bleeding peptic ulcer. PMID- 16469672 TI - Advances and perspectives in the genetics of inflammatory bowel diseases. AB - Several clinical and biological phenotypes define complex diseases such as inflammatory bowel disease (IBD). There is a critical role of the caspase recruitment domain protein 15/nucleotide-binding oligomerization protein 2 dependent (CARD15-NOD2) sensing of bacterial cell wall components in health and disease. The current etiologic model for IBD emphasizes an interaction between susceptibility and modifier genes along with environmental factors. Together, these lead to disease progression. However, further work should clarify the pathophysiological mechanisms leading to IBD and how innate immune signaling confers susceptibility to intestinal inflammation. This is a prerequisite for rational clinical management of IBD. Genetic, functional, serologic testing and development of therapeutics in IBD are discussed. PMID- 16469673 TI - Osteoporosis in patients with inflammatory bowel disease. PMID- 16469674 TI - Selective use of selective nonsteroidal anti-inflammatory drugs in inflammatory bowel disease. PMID- 16469675 TI - Doctors, AGA, and industry: steps toward improving the relationship. PMID- 16469676 TI - The sensitivity of multichannel intraluminal impedance and the pH probe in the evaluation of gastroesophageal reflux in children. AB - BACKGROUND & AIMS: Multichannel intraluminal impedance (MII) has been incorporated into gastroesophageal reflux evaluations in children despite a lack of evidence comparing it to the gold standard pH probe. The aim of this study was to compare these two technologies. METHODS: Twenty-five consecutive, untreated children undergoing pH-MII recording were studied. Sensitivities for the pH probe and MII were defined as: (acid+pH-only episodes)/(acid+non-acid+pH-only episodes) and (acid+non-acid episodes)/(acid+non-acid+pH-only episodes), respectively. Sensitivities were compared using paired t testing. After the analysis was performed, the pH-MII tracings of 25 age-matched children taking acid suppression therapy were compared and sensitivity calculations were performed. RESULTS: In untreated patients, 1845 reflux episodes were detected, and 1702 were detected in treated patients. The mean sensitivities of pH probe and MII in untreated patients were 80.6 +/- 18.2% and 76.1 +/- 13.5%, respectively (P = .41). The mean sensitivities of pH probe and MII in treated patients were 47.2 +/- 36.0% and 80.3 +/- 21.1%, respectively (P = .005). Twenty-five percent of reflux episodes were pH-only episodes. There were no differences in the mean number of acid or non-acid reflux episodes in patients with normal and abnormal pH probe studies. Instead, there were significantly more pH-only episodes in patients with abnormal probe studies. CONCLUSIONS: The sensitivity of MII equals that of the pH probe in untreated patients but is superior to the pH probe in treated patients. pH-only episodes are numerous in pediatrics and should be included in pH-MII analyses. PMID- 16469677 TI - Frozen section analysis of esophageal endoscopic mucosal resection specimens in the real-time management of Barrett's esophagus. AB - BACKGROUND & AIMS: The aim of this study was to assess the validity of frozen section analysis of endoscopic mucosal resection (EMR) specimens from Barrett's esophagus as compared with permanent sections for the detection of neoplasia. Frozen sections help to give immediate feedback for surgical procedures. It has not been determined whether EMR can be adequately interpreted by using frozen sections to aid endoscopists in completely resecting neoplastic lesions. METHODS: EMR specimens from Barrett's esophagus with high-grade dysplasia (HGD) and/or carcinoma were tested by frozen section. Pathologists evaluated EMR specimens for the depth of invasion as well as the appearance of clear margins of resection. The kappa statistic was calculated to assess the degree of agreement between the frozen section and permanent section diagnoses. RESULTS: Twenty-three consecutive patients underwent 30 EMRs with frozen section diagnosis. Frozen section revealed a carcinoma in 7 specimens (23%) and dysplasia in 20 (66%). Permanent sections found carcinoma in 8 specimens (26%), dysplasia in 19 specimens (63%), and normal or nondysplastic Barrett's esophagus in the remainder. The kappa statistic for the depth of invasion of EMR specimens was 0.93 (near perfect agreement). The kappa statistic for the margins of the EMR specimens was 0.80 (excellent agreement). CONCLUSIONS: This study indicated that frozen section analysis of esophageal EMR specimens is valid as compared with permanent section. This technique might allow rapid evaluation about the degree and depth of involvement of cancers. This allows physicians to make decisions regarding further therapy if margins are involved or decrease the use of EMR for histologically benign appearing lesions. PMID- 16469678 TI - Splenic hypofunction and the spectrum of autoimmune and malignant complications in celiac disease. AB - BACKGROUND & AIMS: We investigated the prevalence of functional hyposplenism in autoimmune disorder (AID)-associated and complicated celiac disease (CD). In addition, because the association between hyposplenism and overwhelming infections caused by Streptococcus pneumoniae in patients with CD is well known, we investigated whether immunoglobulin (Ig)M memory B cells, which are responsible for protection against infections by encapsulated bacteria and require the spleen for their generation and/or survival, mirror the reduced splenic function in CD. METHODS: Peripheral blood samples were collected from 73 adult CD patients (27 with AID, 36 without AID, and 10 with CD-related complications including enteropathy-associated T-cell lymphoma, refractory sprue, and ulcerative jejunoileitis). Thirty-four non-CD patients with AID, 35 healthy volunteers, and 29 splenectomized patients also were studied. Counting of erythrocytes with membrane abnormalities (pitted red cells) was used as an indicator of splenic function, and flow cytometry was performed to analyze peripheral blood B cells. RESULTS: A significantly higher risk for hyposplenism was found in AID-associated CD (59%) and complicated CD (80%) than in uncomplicated CD without AID (19%). In AID-associated CD, the degree of splenic function did not correlate to the duration of gluten-free diet. In AID-associated and complicated CD, the frequency of circulating IgM memory B cells was significantly lower than in CD patients without AID or healthy subjects. A significant inverse correlation between IgM memory B cells and pitted red cells was found in all 73 CD patients. CONCLUSIONS: The prevalence of splenic hypofunction is increased in CD with AID and in complicated CD, and is not related to the duration of gluten-free diet. IgM memory B cells are reduced in AID-associated and complicated CD. This defect, which is related to the impairment of splenic function, might predispose hyposplenic CD patients to infections by encapsulated bacteria. PMID- 16469679 TI - Health-related quality of life in patients attending a gastroenterology outpatient clinic: functional disorders versus organic diseases. AB - BACKGROUND & AIMS: Several gastrointestinal (GI) disorders have major effects on health-related quality of life (HRQOL), but there are few direct comparisons between functional GI disorders and organic GI diseases. This study aimed to compare HRQOL between these 2 groups and to assess factors of importance for HRQOL. METHODS: Three hundred ninety-nine consecutive patients attending a GI outpatient clinic completed HRQOL instruments (Short Form 36 [SF-36] and Psychological General Well-Being index [PGWB]) and the Gastrointestinal Symptom Rating Scale (GSRS). For the analyses we divided the patients into 2 diagnostic groups: functional GI disorders (n = 112) and organic GI diseases (n = 287). RESULTS: Compared with norm values on SF-36 and PGWB, both patient groups exhibited profound reductions in HRQOL. After correcting for age, gender, and disease duration, patients with a functional GI disorder had significantly lower scores than patients with an organic GI disease on 6 of 8 SF-36 domains and 5 of 6 PGWB domains. Vitality and anxiety on PGWB, abdominal pain and diarrhea on GSRS, age, and gender independently contributed to the physical component score of SF-36 (adjusted R(2) = 32%). Patients with a functional GI disorder had more severe reflux, abdominal pain, constipation, and indigestion, but the severity of diarrhea did not differ between the groups. HRQOL was reduced with increasing severity of GI symptoms. CONCLUSION: GI disorders have profound effects on HRQOL, and the impact is greater in patients with functional GI disorders as compared with organic GI diseases. The reduction in HRQOL is associated with the severity of both psychological and GI symptoms. PMID- 16469680 TI - Prevalence and mechanism of nonsteroidal anti-inflammatory drug-induced clinical relapse in patients with inflammatory bowel disease. AB - BACKGROUND & AIMS: It has been variably suggested that nonselective NSAIDs and cyclooxygenase (COX)-2 selective inhibitors aggravate or ameliorate clinical disease activity in patients with inflammatory bowel disease. We assessed the effect of these drugs in patients with inflammatory bowel disease (n = 209) and the possible mechanisms. METHODS: First, patients with quiescent Crohn's disease and ulcerative colitis received the non-NSAID analgesic acetaminophen (n = 26) and the conventional NSAIDs naproxen (n = 32), diclofenac (n = 29), and indomethacin (n = 22) for 4 weeks. The Harvey-Bradshaw index was used to define relapse. Second, to assess the mechanism of relapse, intestinal inflammation was quantitated (fecal calprotectin) before and during treatment (20 patients/group) with acetaminophen, naproxen (topical effect, COX-1 and -2 inhibitor), nabumetone (COX-1 and -2 inhibitor), nimesulide (selective COX-2 inhibitor), and low-dose aspirin (selective COX-1 inhibition). RESULTS: Nonselective NSAIDs were associated with a 17%-28% relapse rate within 9 days of ingestion. No patient had an early relapse on acetaminophen, nimesulide, or aspirin, whereas those on naproxen and nabumetone (20%) experienced relapse. These clinical relapses were associated with escalating intestinal inflammatory activity. CONCLUSIONS: NSAID ingestion is associated with frequent and early clinical relapse of quiescent inflammatory bowel disease, and the mechanism appears to be due to dual inhibition of the COX enzymes. Selective COX-2 inhibition with nimesulide and COX 1 inhibition with low-dose aspirin appear to be well-tolerated in the short-term. PMID- 16469682 TI - Aberrant methylation of the eyes absent 4 gene in ulcerative colitis-associated dysplasia. AB - BACKGROUND & AIMS: This study explored the eyes absent 4 (EYA4) gene promoter methylation in noncolitic colorectal tissues and assessed its discrimination for neoplasia in chronic ulcerative colitis (CUC). METHODS: The methylation status of noncolitic specimens was confirmed by direct bisulfite sequencing. Methylation specific polymerase chain reaction (MSP) primers were designed to evaluate colorectal tissues, including 50 noncolitic patients comprising 24 normal epithelia, 14 polyps, and 12 cancers. The assay was tested on tissues from 67 CUC patients including 31 surveillance neoplasia-positive patients and nonneoplastic controls including 22 CUC surveillance-negative and 14 CUC short-disease duration. Remote colonic tissue was included from each of 27 of the 31 CUC neoplasia cases. The expression of EYA4 was quantified in cell lines by use of reverse-transcription polymerase chain reaction. RESULTS: Within noncolitic tissues, bisulfite sequencing showed EYA4 promoter hypermethylation in 80% (8 of 10) of colorectal cancers but in none (0 of 9) of the normal tissues. MSP was positive in 81% (21 of 26) of cancers and polyps and in only 4% (1 of 14) of normal mucosa. In CUC, MSP was positive in 81% (25 of 31) of neoplastic cases but in none (0 of 36) of the nonneoplastic controls. RNA expression was decreased in methylated compared with unmethylated cell lines (P < .001). Treatment with 5-Aza 2'-deoxycytidine (DAC)/Trichostatin (TSA) increased the overall messenger RNA expression (P = .005). CONCLUSIONS: The EYA4 gene promoter is hypermethylated commonly in sporadic and colitic neoplasia and may be associated with gene silencing. EYA4 methylation represents a candidate marker for CUC surveillance. PMID- 16469681 TI - Safety of celecoxib in patients with ulcerative colitis in remission: a randomized, placebo-controlled, pilot study. AB - BACKGROUND & AIMS: The safety of selective cyclooxygenase-2 inhibitors in patients with ulcerative colitis in remission is unknown. METHODS: We performed a placebo-controlled pilot trial to evaluate the safety of celecoxib in patients with ulcerative colitis in remission who had a present or past history of nonspecific arthritis, arthralgia, or other condition amenable to nonsteroidal anti-inflammatory drug therapy. A total of 222 patients with ulcerative colitis in remission were randomized to receive oral celecoxib 200 mg or placebo twice daily for 14 days. Remission was defined as a total Mayo Clinic score of 2 points or less and an endoscopic score of 1 point or less. Disease exacerbation was defined as a total Mayo Clinic score of 5 points or more and an increase in the endoscopic score of 1 point or more. The primary analysis was disease exacerbation through day 14 among patients who underwent randomization, had at least 1 dose of study drug, and had both endoscopy and Mayo Clinic disease activity index scores at the baseline and final assessments. RESULTS: Three percent of patients in the celecoxib group experienced disease exacerbation through day 14, as compared with 4% in the placebo group (P = .719). Eleven percent of patients in each group experienced a bowel-related adverse event (P > .20). CONCLUSIONS: Therapy with celecoxib for up to 14 days did not have a greater relapse rate than placebo in patients with ulcerative colitis in remission who had a present or past history of nonspecific arthritis, arthralgia, or other condition amenable to nonsteroidal anti-inflammatory drug therapy. PMID- 16469684 TI - Histologic spectrum of nonalcoholic fatty liver disease in morbidly obese adolescents. AB - BACKGROUND & AIMS: To characterize the spectrum of nonalcoholic fatty liver disease (NAFLD) in morbidly obese adolescents, we correlated liver histology with clinical features and compared findings with reported adult data. We hypothesized that NAFLD would be less severe as a result of younger age and shorter duration of obesity, but portal inflammation and fibrosis would be more prevalent. METHODS: Cross-sectional study was made of 41 adolescent subjects, 13-19 years old (mean, 16 years), 61% female, 83% non-Hispanic white, mean body mass index 59 kg/m(2), undergoing gastric bypass with liver biopsy. Liver biopsies were graded and staged as proposed by the NASH Clinical Research Network. Data were analyzed by using descriptive statistics, analysis of variance, and Fisher exact tests. RESULTS: Eighty-three percent had NAFLD: 24% steatosis alone, 7% isolated fibrosis with steatosis, 32% nonspecific inflammation and steatosis, and 20% nonalcoholic steatohepatitis (NASH). Twenty-nine percent had fibrosis; none had cirrhosis. Abnormal ALT (P = .05) and AST (P = .01) were more prevalent in NASH. Mean fasting glucose was significantly higher in NASH (P = .05), but prevalence of the metabolic syndrome was not significantly different. CONCLUSIONS: NAFLD was very prevalent in morbidly obese adolescents, but severe NASH was uncommon. In contrast to morbidly obese adults, lobular inflammation, significant ballooning, and perisinusoidal fibrosis were rare, whereas portal inflammation and portal fibrosis were more prevalent, even in those who did not meet criteria for NASH. These findings might support use of a modified scoring system for pediatric NASH. Presence of the metabolic syndrome in morbidly obese adolescents did not distinguish NASH from steatosis alone. PMID- 16469683 TI - Cholangioscopy and cholangioscopic forceps biopsy in patients with indeterminate pancreaticobiliary pathology. AB - BACKGROUND & AIMS: We report the usefulness of cholangioscopy in patients with indeterminate pancreaticobiliary pathology. METHODS: A prospective collection of 62 consecutive patients during a period of 2.5 years who were referred to our tertiary referral center for cholangioscopy for indeterminate strictures suspicious for malignancy were included. Tissue sampling followed cholangioscopic visualization. Biopsies were obtained under direct visualization (cholangioscopy directed) or through the duodenoscope (cholangioscopy-assisted). RESULTS: Sixty two patients had 72 examinations. Forty patients had nondiagnostic sampling before cholangioscopy. Indications were stricture (n = 67: 16 primary sclerosing cholangitis, 51 non-primary sclerosing cholangitis), ductal dilation, or intraductal mass (n = 5). Biopsies were not performed in 19 because cholangioscopy did not identify suspicious lesions. Of the remaining 53 procedures, 29 underwent either cholangioscopy-directed or cholangioscopy assisted biopsy, and 24 had both. Cholangioscopy findings consisted of primary sclerosing cholangitis only (n = 18), benign stricture or inflammatory changes (n = 18), bile duct cancer (n = 14), normal (n = 10), pancreatic cancer (n = 5), and other (n = 7). Fifty-eight patients (94%) had follow-up for a mean of 12.4 months (95% confidence interval, 10.1-14.7). Sixteen of 18 (89%) patients with a final diagnosis of malignancy were detected with cholangioscopy. The 2 missed cancers were intrahepatic cholangiocarcinomas. Overall, sensitivity to detect malignancy by cholangioscopy with and without biopsy was 89%, specificity 96%, positive predictive value 89%, and negative predictive value 96%. CONCLUSIONS: Cholangioscopy with and without biopsy is highly accurate in diagnosing and excluding pancreaticobiliary malignancy in patients with indeterminate strictures. PMID- 16469686 TI - Professionalizing the postdoctoral experience. AB - The first postdoctoral association was launched in the United States 13 years ago. Although postdoctoral associations have made tremendous progress toward improving the lives of postdoctoral fellows, their job is not finished yet. PMID- 16469685 TI - Chronic hepatitis B: a critical appraisal of current approaches to therapy. AB - BACKGROUND & AIMS: Treatment of chronic hepatitis B (CHB) involves a number of complex and controversial issues. Expert opinions may differ from those of practicing hepatologists and gastroenterologists. We aimed to explore this issue further after a critical review of the literature. METHODS: A panel of 14 international experts graded the strength of evidence for 16 statements addressing 3 content areas: patient selection, therapeutic end points, and treatment options. Available data relating to the statements were reviewed critically in 3 small work groups. After discussion of each statement with the entire panel, the experts voted anonymously to accept or reject statements based on the strength of evidence and their experience. A total of 241 members of the American Association for the Study of Liver Diseases (AASLD) responded to the same statements and their responses were compared with those of the experts. A discordant response was defined as a difference of more than 20% in any of the 5 graded levels of response (accept or reject) between the 2 groups. RESULTS: With the exception of 2 statements, the experts' responses were relatively uniform. However, the responses of the AASLD members were discordant from the experts in 12 statements, spanning all 3 content areas. CONCLUSIONS: Several areas of disagreement on the management of CHB exist between experts and AASLD members. Our results indicate a potential knowledge gap among practicing hepatologists. Better educational efforts are needed to meet the challenge of managing this complex disorder in which even expert opinion occasionally may disagree. PMID- 16469687 TI - Creationism and evolution: it's the American way. AB - The recent ruling in the Kitzmiller v. Dover court case that intelligent design is a form of religion and cannot be taught alongside evolution in science classes in US public schools garnered worldwide attention. But why is the antievolution movement so powerful in the United States? PMID- 16469688 TI - Sex steroids and the free hormone hypothesis. PMID- 16469690 TI - Rejuvenating exhausted T cells during chronic viral infection. AB - In a recent paper in Nature, show that the immunoreceptor PD-1 is upregulated by "exhausted" T cells during the chronic phase of viral infection in mice. Remarkably, blocking the interaction between PD-1 and its ligand, PD-L1, reactivates these T cells and reduces viral load. PMID- 16469691 TI - To kill but not be killed: a delicate balance. AB - Before launching a missile, it is necessary to design an efficient safety net for self-protection. In this issue of Cell, Ellermeier et al. (2006) describe the mechanism underlying a biological safety net for the soil bacterium Bacillus subtilis. This bacterium protects itself from a toxic protein it secretes by upregulating an immunity protein, which it does by sequestering a transcriptional repressor at the plasma membrane. PMID- 16469692 TI - How the ER stays in shape. AB - Understanding the molecular mechanisms that control the architecture of organelles is an area of intense study. In this issue of Cell, Voeltz et al. (2006) report that two membrane proteins, Rtn4a/NogoA and DP1/Yop1p, are responsible for the generation of tubular morphology in the endoplasmic reticulum (ER). The unusual membrane topology of these proteins may directly contribute to ER curvature. PMID- 16469693 TI - Inflammation and sex steroid receptors: a motif for change. AB - Homeostasis in reproductive tissues requires integration of hormonal and inflammatory signals. In this issue of Cell, Zhu et al. (2006) discover that proinflammatory signals switch repressed steroid hormone receptors into transcriptional activators by targeting TAB2, an adaptor protein that tethers corepressors. These findings have implications for the treatment of endocrine resistant cancers. PMID- 16469694 TI - MEDEA takes control of its own imprinting. AB - Genomic imprinting is an essential epigenetic process that controls the size of seeds in flowering plants. In Arabidopsis, DEMETER activates the maternal copy of the imprinted MEDEA Polycomb gene. In this issue of Cell, Gehring et al. (2006) demonstrate that this activation involves DNA demethylation of MEDEA by DEMETER. Remarkably, they also find that silencing of the paternal MEDEA allele is independent of DNA methylation and is controlled by maternal expression of MEDEA itself. PMID- 16469695 TI - TOR signaling in growth and metabolism. AB - The target of rapamycin (TOR) is a conserved Ser/Thr kinase that regulates cell growth and metabolism in response to environmental cues. Here, highlighting contributions from studies in model organisms, we review mammalian TOR complexes and the signaling branches they mediate. TOR is part of two distinct multiprotein complexes, TOR complex 1 (TORC1), which is sensitive to rapamycin, and TORC2, which is not. The physiological consequences of mammalian TORC1 dysregulation suggest that inhibitors of mammalian TOR may be useful in the treatment of cancer, cardiovascular disease, autoimmunity, and metabolic disorders. PMID- 16469696 TI - Cryo-EM reconstruction of dengue virus in complex with the carbohydrate recognition domain of DC-SIGN. AB - Dengue virus (DENV) is a significant human pathogen that causes millions of infections and results in about 24,000 deaths each year. Dendritic cell-specific ICAM3 grabbing nonintegrin (DC-SIGN), abundant in immature dendritic cells, was previously reported as being an ancillary receptor interacting with the surface of DENV. The structure of DENV in complex with the carbohydrate recognition domain (CRD) of DC-SIGN was determined by cryo-electron microscopy at 25 A resolution. One CRD monomer was found to bind to two glycosylation sites at Asn67 of two neighboring glycoproteins in each icosahedral asymmetric unit, leaving the third Asn67 residue vacant. The vacancy at the third Asn67 site is a result of the nonequivalence of the glycoprotein environments, leaving space for the primary receptor binding to domain III of E. The use of carbohydrate moieties for receptor binding sites suggests a mechanism for avoiding immune surveillance. PMID- 16469697 TI - DEMETER DNA glycosylase establishes MEDEA polycomb gene self-imprinting by allele specific demethylation. AB - MEDEA (MEA) is an Arabidopsis Polycomb group gene that is imprinted in the endosperm. The maternal allele is expressed and the paternal allele is silent. MEA is controlled by DEMETER (DME), a DNA glycosylase required to activate MEA expression, and METHYLTRANSFERASE I (MET1), which maintains CG methylation at the MEA locus. Here we show that DME is responsible for endosperm maternal-allele specific hypomethylation at the MEA gene. DME can excise 5-methylcytosine in vitro and when expressed in E. coli. Abasic sites opposite 5-methylcytosine inhibit DME activity and might prevent DME from generating double-stranded DNA breaks. Unexpectedly, paternal-allele silencing is not controlled by DNA methylation. Rather, Polycomb group proteins that are expressed from the maternal genome, including MEA, control paternal MEA silencing. Thus, DME establishes MEA imprinting by removing 5-methylcytosine to activate the maternal allele. MEA imprinting is subsequently maintained in the endosperm by maternal MEA silencing the paternal allele. PMID- 16469698 TI - Structural basis for bacterial transcription-coupled DNA repair. AB - Coupling of transcription and DNA repair in bacteria is mediated by transcription repair coupling factor (TRCF, the product of the mfd gene), which removes transcription elongation complexes stalled at DNA lesions and recruits the nucleotide excision repair machinery to the site. Here we describe the 3.2 A resolution X-ray crystal structure of Escherichia coli TRCF. The structure consists of a compact arrangement of eight domains, including a translocation module similar to the SF2 ATPase RecG, and a region of structural similarity to UvrB. Biochemical and genetic experiments establish that another domain with structural similarity to the Tudor-like domain of the transcription elongation factor NusG plays a critical role in TRCF/RNA polymerase interactions. Comparison with the translocation module of RecG as well as other structural features indicate that TRCF function involves large-scale conformational changes. These data, along with a structural model for the interaction of TRCF with the transcription elongation complex, provide mechanistic insights into TRCF function. PMID- 16469699 TI - Bruno acts as a dual repressor of oskar translation, promoting mRNA oligomerization and formation of silencing particles. AB - Prior to reaching the posterior pole of the Drosophila oocyte, oskar mRNA is translationally silenced by Bruno binding to BREs in the 3' untranslated region. The eIF4E binding protein Cup interacts with Bruno and inhibits oskar translation. Validating current models, we directly demonstrate the mechanism proposed for Cup-mediated repression: inhibition of small ribosomal subunit recruitment to oskar mRNA. However, 43S complex recruitment remains inhibited in the absence of functional Cup, uncovering a second Bruno-dependent silencing mechanism. This mechanism involves mRNA oligomerization and formation of large (50S-80S) silencing particles that cannot be accessed by ribosomes. Bruno dependent mRNA oligomerization into silencing particles emerges as a mode of translational control that may be particularly suited to coupling with mRNA transport. PMID- 16469700 TI - A dynamically localized protease complex and a polar specificity factor control a cell cycle master regulator. AB - Regulated proteolysis is essential for cell cycle progression in both prokaryotes and eukaryotes. We show here that the ClpXP protease, responsible for the degradation of multiple bacterial proteins, is dynamically localized to specific cellular positions in Caulobacter where it degrades colocalized proteins. The CtrA cell cycle master regulator, that must be cleared from the Caulobacter cell to allow the initiation of chromosome replication, interacts with the ClpXP protease at the cell pole where it is degraded. We have identified a novel, conserved protein, RcdA, that forms a complex with CtrA and ClpX in the cell. RcdA is required for CtrA polar localization and degradation by ClpXP. The localization pattern of RcdA is coincident with and dependent upon ClpX localization. Thus, a dynamically localized ClpXP proteolysis complex in concert with a cytoplasmic factor provides temporal and spatial specificity to protein degradation during a bacterial cell cycle. PMID- 16469701 TI - A three-protein signaling pathway governing immunity to a bacterial cannibalism toxin. AB - We describe a three-protein signal-transduction pathway that governs immunity to a protein toxin involved in cannibalism by the spore-forming bacterium Bacillus subtilis. Cells of B. subtilis enter the pathway to sporulate under conditions of nutrient limitation but delay becoming committed to spore formation by killing nonsporulating siblings and feeding on the dead cells. Killing is mediated by the exported toxic protein SdpC. We report that extracellular SdpC induces the synthesis of an immunity protein, SdpI, that protects toxin-producing cells from being killed. SdpI, a polytopic membrane protein, is encoded by a two-gene operon under sporulation control that contains the gene for an autorepressor, SdpR. The autorepressor binds to and blocks the promoter for the operon. Evidence indicates that SdpI is also a signal-transduction protein that responds to the SdpC toxin by sequestering the SdpR autorepressor at the membrane. Sequestration relieves repression and stimulates synthesis of immunity protein. PMID- 16469702 TI - A receptor-modifying deamidase in complex with a signaling phosphatase reveals reciprocal regulation. AB - Signal transduction underlying bacterial chemotaxis involves excitatory phosphorylation and feedback control through deamidation and methylation of sensory receptors. The structure of a complex between the signal-terminating phosphatase, CheC, and the receptor-modifying deamidase, CheD, reveals how CheC mimics receptor substrates to inhibit CheD and how CheD stimulates CheC phosphatase activity. CheD resembles other cysteine deamidases from bacterial pathogens that inactivate host Rho-GTPases. CheD not only deamidates receptor glutamine residues contained within a conserved structural motif but also hydrolyzes glutamyl-methyl-esters at select regulatory positions. Substituting Gln into the receptor motif of CheC turns the inhibitor into a CheD substrate. Phospho-CheY, the intracellular signal and CheC target, stabilizes the CheC:CheD complex and reduces availability of CheD. A point mutation that dissociates CheC from CheD impairs chemotaxis in vivo. Thus, CheC incorporates an element of an upstream receptor to influence both its own effect on receptor output and that of its binding partner, CheD. PMID- 16469703 TI - A class of membrane proteins shaping the tubular endoplasmic reticulum. AB - How is the characteristic shape of a membrane bound organelle achieved? We have used an in vitro system to address the mechanism by which the tubular network of the endoplasmic reticulum (ER) is generated and maintained. Based on the inhibitory effect of sulfhydryl reagents and antibodies, network formation in vitro requires the integral membrane protein Rtn4a/NogoA, a member of the ubiquitous reticulon family. Both in yeast and mammalian cells, the reticulons are largely restricted to the tubular ER and are excluded from the continuous sheets of the nuclear envelope and peripheral ER. Upon overexpression, the reticulons form tubular membrane structures. The reticulons interact with DP1/Yop1p, a conserved integral membrane protein that also localizes to the tubular ER. These proteins share an unusual hairpin topology in the membrane. The simultaneous absence of the reticulons and Yop1p in S. cerevisiae results in disrupted tubular ER. We propose that these "morphogenic" proteins partition into and stabilize highly curved ER membrane tubules. PMID- 16469704 TI - Endoplasmic reticulum stress activates cleavage of CREBH to induce a systemic inflammatory response. AB - Regulated intramembrane proteolysis (RIP) of endoplasmic reticulum (ER) membrane anchored transcription factors is known to maintain sterol homeostasis and to mediate the unfolded protein response (UPR). Here, we identified CREBH as a RIP regulated liver-specific transcription factor that is cleaved upon ER stress and required to activate expression of acute phase response (APR) genes. Proinflammatory cytokines increase expression of ER membrane-anchored CREBH. In response to ER stress, CREBH is cleaved by site-1 and site-2 proteases to liberate an amino-terminal fragment that transits to the nucleus to activate transcription of the genes encoding serum amyloid P-component (SAP) and C reactive protein (CRP). Proinflammatory cytokines and lipopolysaccharide activate the UPR and induce cleavage of CREBH in the liver in vivo. Together, our studies delineate a molecular mechanism for activation of an ER-localized transcription factor, CREBH, and reveal an unprecedented link by which ER stress initiates an acute inflammatory response. PMID- 16469705 TI - The E3 ubiquitin ligase itch couples JNK activation to TNFalpha-induced cell death by inducing c-FLIP(L) turnover. AB - The proinflammatory cytokine tumor necrosis factor (TNF) alpha signals both cell survival and death. The biological outcome of TNFalpha treatment is determined by the balance between NF-kappaB and Jun kinase (JNK) signaling; NF-kappaB promotes survival, whereas JNK enhances cell death. Critically, identity of a JNK substrate that promotes TNFalpha-induced apoptosis has been outstanding. Here we show that TNFalpha-mediated JNK activation accelerates turnover of the NF-kappaB induced antiapoptotic protein c-FLIP, an inhibitor of caspase-8. This is not due to direct c-FLIP phosphorylation but depends on JNK-mediated phosphorylation and activation of the E3 ubiquitin ligase Itch, which specifically ubiquitinates c FLIP and induces its proteasomal degradation. JNK1 or Itch deficiency or treatment with a JNK inhibitor renders mice resistant in three distinct models of TNFalpha-induced acute liver failure, and cells from these mice do not display inducible c-FLIP(L) ubiquitination and degradation. Thus, JNK antagonizes NF kappaB during TNFalpha signaling by promoting the proteasomal elimination of c FLIP(L). PMID- 16469706 TI - Macrophage/cancer cell interactions mediate hormone resistance by a nuclear receptor derepression pathway. AB - Defining the precise molecular strategies that coordinate patterns of transcriptional responses to specific signals is central for understanding normal development and homeostasis as well as the pathogenesis of hormone-dependent cancers. Here we report specific prostate cancer cell/macrophage interactions that mediate a switch in function of selective androgen receptor antagonists/modulators (SARMs) from repression to activation in vivo. This is based on an evolutionarily conserved receptor N-terminal L/HX7LL motif, selectively present in sex steroid receptors, that causes recruitment of TAB2 as a component of an N-CoR corepressor complex. TAB2 acts as a sensor for inflammatory signals by serving as a molecular beacon for recruitment of MEKK1, which in turn mediates dismissal of the N-CoR/HDAC complex and permits derepression of androgen and estrogen receptor target genes. Surprisingly, this conserved sensor strategy may have arisen to mediate reversal of sex steroid dependent repression of a limited cohort of target genes in response to inflammatory signals, linking inflammatory and nuclear receptor ligand responses to essential reproductive functions. PMID- 16469708 TI - [Single nucleotide polymorphisms in NOS3 A-922G, T-786C and G894T: a correlation study of the distribution of their allelic combinations with hypertension in chinese Han population]. AB - To study single nucleotide polymorphisms (SNP) in A-922G, T-786C and G894T of endothelial nitric oxide synthase (NOS3) and to correlate the distribution of their allelic combinations with hypertension in Chinese Han nationality population, genomic DNA was isolated from venous blood leukocytes from 192 unrelated patients with hypertension (95 females and 97 males) and 122 healthy unrelated individuals (46 females and 76 males) as controls. SNPs of NOS3 A-922G, T-786C and G894T were genotyped by allele-specific primer (ASP) PCR. The distribution of genotype combinations of three SNPs was determined by clustering analysis. There were no difference in allele genotype distribution frequency and haplotype frequency of NOS3 G894T, NOS3 A-922G and NOS3 T-786C between the essential hypertension group and the healthy population (P>0.05). According to sex stratification, no association between essential hypertension and SNP of NOS3 A-922G,NOS3 T-786C or NOS3 G894T has been found in either the male subgroup or the female subgroup. In respect of allele genotype combination frequency in the natural distribution of NOS3 A-922 G, NOS3 T-786C and NOS3 G894T SNP, there was significant difference only in the allele genotype combination frequency of NOS3 G894G+A-922G+T-786T between the hypertension group and the healthy group (P<0.05, Chi2=4.5944). According to sex stratification, there were no significant difference in all above allele genotype combination frequency in three sites of NOS3 SNP between the hypertension male subgroup and the healthy male subgroup (P>0.05). There was significant difference in the allele combination frequency of NOS3 G894G +A-922G+T-786C between the hypertension female subgroup and the healthy female subgroup(P<001, Chi2=8.502). There was no association of SNP in NOS3 A-922G, NOS3 T-786C or NOS3 G894T with hypertension in the Chinese Han nationality population, nor was there a sex difference. The combination frequency of allele NOS3 G894G + A-922G + T-786C in the hypertension female subgroup was much lower than that in the healthy female subgroup, suggesting that female population with this combination genotype may be less susceptible to hypertension. PMID- 16469707 TI - p120-catenin mediates inflammatory responses in the skin. AB - Although p120-catenin regulates adherens junction (AJ) stability in cultured cells, genetic studies in lower eukaryotes have not revealed a role for this protein in vivo. Using conditional targeting in mice, we show that p120 null neonatal epidermis exhibits reduced intercellular AJ components but no overt disruption in barrier function or intercellular adhesion. As the mice age, however, they display epidermal hyperplasia and chronic inflammation, typified by hair degeneration and loss of body fat. Using skin engraftments and anti inflammatory drugs, we show that these features are not attributable to reductions in junctional cadherins and catenins, but rather NFkB activation. Both in vivo and in vitro, p120 null epidermal cells activate nuclear NFkB, triggering a cascade of proinflammatory NFkB targets. Although the underlying mechanism is likely complex, we show that p120 affects NFkB activation and immune homeostasis in part through regulation of Rho GTPases. These findings provide important new insights into p120 function. PMID- 16469710 TI - [Genetic polymorphism of penta e locus in four Chinese nationalities]. AB - To study the genetic polymorphism of Penta E locus in four Chinese nationalities using home made reagent kits, DNA samples were obtained from about 400 unrelated peoples of four different Chinese nationalities. As a result, we found 20 alleles in the four nationalities with frequencies ranging from 0.0048 to 0.2396. The genotype frequencies of Penta E locus met Hardy-Weinberg equilibrium. It proved that Penta E locus was a high polymorphic STR genetic marker and was valuable for forensic science. PMID- 16469709 TI - [Genetic analysis of the UGT1A1 gene mutation sites in a Chinese family suffered from Gilbert's syndrome]. AB - To learn the variation in the gene for UGT1A1 enzyme, the genetic mechanism in a Chinese Han nationality family suffered from Gilbert's syndrome was studied. At first, genomic DNA from peripheral blood of the sufferer in this family was used for amplifying all of the five exons of the UGT1A1 gene by PCR, and then direct sequencing of the PCR product was applied to analyze gene mutation. The results showed that there existed a G-->A homozygous transition at nucleotide 211 leading the substitution of arginine for glycine at position 71 of corresponding protein product (G71R) and a T-->G homozygous transition at nucleotide 1456 leading the substitution of aspartic acid for tyrosine at position 486 of corresponding protein product (Y486D). No mutation was detected in promoter region and the splicing junction sites. The relevant mutation sites of the other family members were sequenced and identified to be heterozygous in the two above-mentioned mutation sites and in the TA repeat mutation in the promoter region. Furthermore, fresh blood samples were collected from all of the members to detect the serum bilirubin levels to determine the sufferer. The result was consistent with the mutation analysis. It could thus be inferred that this family was caused by mutation in the open reading frame of the gene UGT1A1. PMID- 16469711 TI - [Knockdown of PLK1 mRNA by special siRNA]. AB - To study knockdown effect of small interfering RNA (siRNA) to PLK1 (Polo-like kinase 1) mRNA in colorectal cancer cell line SW480 and its mitosis and growth was changed. Ten special siRNA molecules were designed targeting different sites of PLK1 mRNA sequence and chemically synthesized. The siRNA molecules were transfected into SW480 by Oligofectamine. The gene mRNA level was assayed by Real Time PCR. The changes of PLK1 protein, SW480 cell cycle and survival percentage was checked by Western-blot, Flow cytometry and Cell counter assays respectively. All 10 siRNA molecules knocked PLK1 mRNA down in different level. Of them P1, P4 and P9 showed over 80% knockdown efficiency and the others had more than 20% knockdown efficiency to PLK1 mRNA. The best knockdown effect over 95% of all groups was at 25 nmol/L of a mixture with P1, P4 and P9 siRNA equally. In this situation the protein was very less and the cells were blocked at G2 phase of cell cycle. After 72 h cell survival percentages were consistent with PKL1 mRNA level change by siRNA gradient concentration. The results showed that siRNA targeting PLK1 mRNA had effectively knocked PLK1 mRNA down in SW480 cell line. And a blended siRNAs held the best knockdown effect. The cell was blocked on the mitosis and growth. PMID- 16469712 TI - [Genome-wide search for linkage to attention deficit hyperactivity disorder (ADHD) on the X chromosome]. AB - Attention deficit hyperactivity disorder (ADHD) is the most common childhood onset behavioral. Boys are more often affected than girls. Family, twin and adoption studies have supported a strong genetic basis. The etiology of this disorder is not clear. Molecular genetic and pharmacological studies suggest the involvement of dopaminergic and noradrenergic neurotransmitter systems in ADHD, e.g , Several reports have found association between ADHD and the dopamine receptor gene DRD-4.the dopamine transporter gene DAT1, and the catechol-o methyltransferase. Our previous studies showed an association between ADHD and the DXS7 locus, which is located in closely linked to the MAO gene, and MAOA gene on chromosome X. To test this hypothesis, we used the genome scan for a predisposing locus on chromosome X to ADHD. We used the tramsmission/disequilibrium test (TDT) to test for linkage between a VNTR polymorphism at the 48 markers of chromosome X and DSM-III-R oliagnosed ADHD in 84 nuclear families of the Chinese population. The TDT analysis revealed linkage between ADHD and the DXS1214(TDT: Chi2=18.1, df=7, P<0.01), DXS8102(TDT: Chi2=7.9, df=3, P<0.05), DXS1068(TDT: Chi2=21.9, df=9, P<0.01), DXS8015(TDT: Chi2=14.6, df=7, P<0.05), DXS1059(TDT: Chi2=27.8, df=10, P<0.01) and DXS8088(TDT: Chi2=20.4, df=3, P<0.01).The data showed that susceptibility loci might reside in chromosome Xp11.4-Xp21 and Xq23 for ADHD. PMID- 16469713 TI - [Molecular cloning and functional analysis of bovine mammary gland matrix attachment regions]. AB - The bovine genomic DNA was extracted from bovine blood, then bovine mammary gland matrix attachment region (BMARs) was cloned using a pair of primers, which were designed based on the related sequences in GenBank through bio-software Primer5.0 and Vector7.0. Upon preliminary analysis with bio-software, BMARs was TA cloned into PMD-18 T vector. By means of adding Kpn2 I and Xho I to 5' upstream of sensitive and antisensitive primers respectively, expressing vector BE was constructed after BMAR was cloned into the downstream of the reporter gene in pEGFP-C1. Bovine ear fibroblast cells were transfected by expressing vector BE with Lipofectamine.Compared with control bovine ear fibroblast cells transfected with pEGFP-C1, the effect of cloned BMR was apparent in dispelling position effect and enhancing gene expression. PMID- 16469714 TI - [A new polymorphic site of ESR gene in pigs]. AB - ESR (Estrogen Receptor) gene has been determined to be one of the major genes affecting phenotype of litter size without any genetic negative correlation to growth and carcass trait. Recent studies mainly focused on the PvuII enzyme site found by Rothschild in 1996. An optimized standard PCR-SSCP protocol was employed to detect the exon 7 of ESR gene. The results showed that there were a new mutation site and three genotypes, which may be used as a new DNA marker site to further research. PMID- 16469716 TI - [Separation and characterization of mitochondrial COI pseudogenes in Scylla paramamosain]. AB - Mitochondrial DNA marker has been applied in genetic structure and phylogenetic analysis, but the differences between nuclear mitochondrial pseudogenes (Numts) and mitochondrial DNA complicate phylogenetic analysis. Using the universal and special primers, 34 pseudogene sequences (Numts) and 5 real mtDNA COI sequences were amplified respectively from 34 individuals of Scylla paramamosain from the coast of southeast China. Twenty-nine haplotypes were identified and two groups (Group I and Group II) were determined according to the homology of the 34 pseudogene sequences. Two insertions and an 8-bp deletion were found in some sequences of Group I, but no indel (insertion and deletion) was found in Group II. Compared to mtDNA COI sequences, the two groups had their conservative sites respectively. At least two independent nuclear transfer events are required to explain the codon position biases revealed through the pairwise comparisons of the 34 Numts, one for Group I and one for Group II. The results showed the necessity of great care in interpreting PCR-generated mtDNA sequences using universal primers when used in population or evolutionary studies in genus Scylla. PMID- 16469715 TI - [Study of single nucleotide polymorphism of A-FABP gene and its association with fatness traits in chicken]. AB - In this experiment, Beijing-You chicken was used to detect single nucleotide polymorphisms (SNPs) in A-FBAP gene and to study the correlation between its genotype and the trait of fat accumulation. The first exon of the gene was amplified by one pair of primers, and SNPs were detected by the technique of single strand conformation polymorphism (SSCP) and finally confirmed by sequencing. Results of analysis of variance showed that a significant difference existed among abdominal fat percentage, subcutaneous fat thickness and intramuscular fat contents in breast musculature with different genotypes (P<0.01). It implied that A-FABP gene could be a major effector gene or could be linked to gene(s) which significantly affect fat metabolism in chicken. PMID- 16469717 TI - [Genetic analysis and RAPD marker of the genes for brachytic stem trait in soybean]. AB - Three crosses between NG94-156 (brachytic stem) and three varieties (normal stem) were made, and F2 segregative population and two recombined inbred line populations(F(7:8)) were obtained. Genetic analysis indicated that the brachytic stem of NG94-156 was controlled by two duplicate recessive genes. In searching for RAPD marker linked to the genes controlling brachytic stem, 260 RAPD primers were applied to screen four parents of three combinations and RIL. Polymorphic bands revealed by the primer S-506 exhibited the best repeatability among all primers. Linkage analysis indicated the genetic distance between S-506(1600) and brachytic stem gene was 6.94 cM. PMID- 16469718 TI - [Molecular cloning of a novel low-molecular-weight glutenin subunit gene from wheat variety "Chuanmai 42"]. AB - Using PCR-amplification method, the coding sequences of a novel low-molecular weight glutenin (LMW-GS) gene, LMWCM42-1, was isolated from the genomic DNA of wheat (Triticum aestivum L.) variety 'Chuanmai 42'. The full coding region of this gene consisted of 846 nucleotides, and encoded a protein with 281 amino acids. LMWCM42-1 is a typical low-molecular-weight glutenin subunit (LMW-GS) gene. In spite of high similarity with the known LMW-GSs, LMWCM42-1 was quite different from the known LMW-GS in the N-terminal, central repetitive domain and C-terminal domain. It was found that LMWCM42-1 was located within the Glu-D3 locus by phylogenetic analysis. PMID- 16469719 TI - [Genetic transformation of peach immature cotyledons with its antisenes ACO gene]. AB - Genetic transformation of peach immature cotyledons with its ACO antisense gene was studied by using particle bombardment method through Agrobacterium tumefaciens. Kanr shoots and Kanr plantlet were obtained. The plantet with ACO antisenes gene through Agrobacterium tumefaciens was obtained by micrografting technique and survived for nearly one month. The results of the PCR, PCR southern, genomes southern hybridization analysis and GUS color reaction of some Kanr materials showed in some degree that the peach ACO antisens gene was integrated into peach genomes. PMID- 16469720 TI - [pBR322-Red mediated gene knockin, sites and expression in E. coli chromosome]. AB - Genes lacZ, lacY and lacA in the lac opron of E. coli chromosome were respectively substituted with gene luc by using plasmid pBR322-Red, selection counterselection system kan/sacB and various strategies of Red homologous recombination including Red mediated linearized double-stranded DNA homologous recombination and Red mediated recombineering with overlapping single stranded DNA oligonucleotides. Then, a series of new strains, CWL2, CWL4 and CWL6, were constructed and we found that they can express protein Luc efficiently. To further study the expression of exogenous genes at the site of lacZ, we have constructed a strain named CWD1 by knockin the cholera toxin B subunit(ctxb) gene at the lacZ site, then we found that CWD1 can express protein CTB efficiently and CTB was secreted out of the cell. So we assured that the sites of structure genes in the lac operon of Escherichia coli chromosome were suitable for expressing foreign genes. PMID- 16469721 TI - [A new method of molecular marker--RMAPD]. AB - Random microsatellite amplify polymorphic DNA (RMAPD) is a new method of molecular marker. It can reveal genome DNA polymorphisms of organism only when random primer and microsatellite forward or reverse primer are combined together as a pair of primers by the PCR reaction system with Taq DNA polymerase, MgCl2, dNTPs and contemplate DNA. The core question of RMAPD is validity of the primers used. A lot of experiments of RMAPD in 69 Xinong Saanen dairy goats showed that RMAPD primers were effective. Comparison of RMAPD, microsatellite and RAPD markers demonstrated that RMAPD was different from each other in primers, amplification protocol and repetition. RMAPD is not equal to RAPD, but a extendable RAPD. Therefore, RMAPD is regarded as a new method of molecular marker. As it has many characteristics of DNA marker, RMAPD has a widest potential application in animal breeding and genetics, such as analysis of genetic structure and relationship and MAS. PMID- 16469722 TI - [Rapid determination of transient expression characteristics of drought- and dehydration-inducible promoters from plants]. AB - The selection of suitable inducible-promoters is one of the most important chains in transgenic researches for plant tolerances to drought, dehydration and other abiotic stresses. Based on laboratory researches in the past years, it was set up a set of methods that could be used to rapidly determine the transient expression characteristics of drought- and dehydration-inducible promoters by using whole barley seedling and plant leaves in vitro as bombardment objects. Promoters Dhn4s, Dhn8s, HVA1s, Rab16Bj and wsi18j isolated from barley and rice were able to express GFP in the leaves of barley, wheat, rice, sorghum and fern after drying treatment, but not in laves of mungbean and tomato. The qualitative expression of HVA1s and wsi18j in different organs and tissues of barley was identified. A method for quantitative analysis of promoter transient expression was established by means of GFP foci/GUS foci counting or GUS activity/XYN activity test. Finally, the value and prospect of the methodology were discussed in terms of its application to the analysis of plant promoters inducible by environmental factors. PMID- 16469724 TI - [Application of DNA-related techniques in avian molecular phylogeny]. AB - The DNA techniques most commonly used in avian molecular phylogeny include DNA hybridization, RFLP and DNA sequence analysis, among which DNA sequence analysis is supposed to be the most effective and reliable. DNA hybridization techniques have been widely used in aves, based on which a new avian classification system was born. In avian RFLP analyses, mtDNA are widely used as target sequences. Mitochondrial DNA genes are the most frequently used in avian molecular phylogeny. Although mtDNA phylogenies are likely to be correct in many cases, use of mtDNA sequences can be problematic with such constraints as unilateral inheritance, multiple substitutions, saturations at the third-coded sites, strong bias in base composition and probable nuclear pseudogenes of mtDNA sequences. Although bias are still on the mtDNA sequences, more and more authors turn to nuclear DNA sequences and prefer to a combination of mtDNA and nuclear DNA sequences. And single-copy nuclear DNA receives the most favor. scnDNA introns can perform well in recovering relationships among intermediate to even distantly related congeneric species. scnDNA exons can be used in avian higher ranks. With the exception of molecular markers' own problems including variable rates of nucleotide site evolution, gene hybridization, gene horizontal transfer and lineage sorting, avian molecular phylogeny also faces methodological problems, such as molecular markers selection, taxon sampling and data processing. More attention should be paid to the standardization of methods, not to the new molecular markers. PMID- 16469723 TI - [Rational evolutionary design and modification of gene: a short-path of direct evolution]. AB - Directed evolution is a powerful means in academic study and industrial application to modified and functionally improved proteins. There have been significant and impressive advances using the artificial method. Rational Evolutionary Design utilizes structural and sequence alignment information to create new genes and proteins. Rational Evolutionary Design has recently emerged as an attractive approach for studying function of proteins. The review mainly introduced four aspects: preferred codon usage, structural and sequence alignment, key domain analysis, site-directed recombination. PMID- 16469725 TI - [Advances in researches on genetic markers of 2n gametes]. AB - 2n gametes are the result of meiotic mutation during micro- and mega-sporogenesis that bear the sporophytic rather than the gametophytic chromosome number. This paper reviewed the genetic markers including the morphologic, cytological, isozymes and DNA markers, which have been employed in the generation, inheritance, heterozygosity and marker-assisted breeding of 2n gametes based on the frequency of large pollen grains, cytological analyses, unexpected occurrence of polyploidy progeny and the associations between parents and progenies. Based on a better understanding of the genetic control and meiotic mutations responsible for 2n gametes formation, this could open up new prospects in the use of 2n gametes for plant breeding. It should be possible to use marker- assisted selection of superior 2n gametes for polyloid breeding. PMID- 16469726 TI - [Key enzymes in starch synthesis in plants]. AB - Starch, the most common form of stored carbon in plants, is both the major food source for mankind and important raw material for many industries. It is composed of two types of alpha-1,4-linked glucan polymer: essentially unbranched amylose and regularly branched amylopectin, and synthesized in photosynthetic and non photosynthetic organs. Starch is synthesized via four committed enzyme steps: ADP Glc pyrophosphorylase, which synthesizes sugar nucleotide precursors; starch synthase, which extends the alpha-1,4-linked glucan chains using ADP-Glc; starch branching enzymes, which introduce alpha-1,6 branch points to form amylopectin; and starch debranching enzymes, which hydrolyze alpha-1,6 branches in glucans. In this paper, recent advances in biochemical characterizations and gene engineering concerning these enzymes were reviewed, and the achievements in gene engineering involved in manipulation of starch amount and quality were also cited. PMID- 16469727 TI - [Approaches for SNP genotyping]. AB - With the completion of the Human Genome Project (HGP), typing single nucleotide polymorphisms (SNP) has become one of the main tasks in the post-genome era. Consequently, a robust, flexible and cost-effective technique for SNP typing is essential to analyze a large number of SNPs. The latest genotyping technologies and the relative detection platforms were introduced systematically. The principle of SNP typing was described in detail in respect of allele-specific hybridization, restriction enzyme digestion, primer extension and oligonucleotide ligation assay, as well as the genotyping platforms of plate readers, genechips, bead array and mass spectrometry. Moreover, the way to the high-throughput genotyping in the future was briefly discussed. PMID- 16469728 TI - The sour side of sweet. PMID- 16469729 TI - Mucking with metabolism. PMID- 16469730 TI - When good Cdk5 turns bad. AB - The cyclin-dependent kinase-5 (Cdk5) is critical to normal mammalian development and has been implicated in synaptic plasticity, learning, and memory in the adult brain. But Cdk-5 activity has also been linked to neurodegenerative diseases. Could a single protein have opposing effects? A new study shows that production of a neuronal protein capable of regulating Cdk-5 activity can turn Cdk-5 from "good" to "bad." The findings may have implications for the development and treatment of conditions like Alzheimer's disease. PMID- 16469731 TI - Olfactory loss in aging. AB - Olfactory loss is a common age-related complaint that may be caused by changes in the anatomy of the structures required for olfaction (for example, loss of olfactory receptor cells) or in the environment surrounding the receptor cell (for example, altered nasal mucus composition). However, aging, as well as age related diseases and medications, may also alter the distribution, density, or function of specific receptor proteins, ion channels, or signaling molecules that affect the ability of neural elements throughout the olfactory pathway to signal and process odorant information. Although a great deal has been learned about the prevalence and nature of age-related olfactory loss, we are just beginning to explore avenues to prevent or alleviate this sensory deficit. Some studies suggest that, rather than being a necessary outcome of aging, age-associated factors such as chronic diseases, medications, and dental and sinus problems are the primary culprits in causing olfactory impairment. This idea suggests optimism in that, as we address these other age-related health issues, the prevalence of olfactory loss will lessen as well. PMID- 16469732 TI - Modulation of Nav1.5 channel function by an alternatively spliced sequence in the DII/DIII linker region. AB - In the present study, we identified a novel splice variant of the human cardiac Na(+) channel Na(v)1.5 (Na(v)1.5d), in which a 40-amino acid sequence of the DII/DIII intracellular linker is missing due to a partial deletion of exon 17. Expression of Na(v)1.5d occurred in embryonic and adult hearts of either sex, indicating that the respective alternative splicing is neither age-dependent nor gender-specific. In contrast, Na(v)1.5d was not detected in the mouse heart, indicating that alternative splicing of Na(v)1.5 is species-dependent. In HEK293 cells, splice variant Na(v)1.5d generated voltage-dependent Na(+) currents that were markedly reduced compared with wild-type Na(v)1.5. Experiments with mexiletine and 8-bromo-cyclic AMP suggested that the trafficking of Na(v)1.5d channels was not impaired. However, single-channel recordings showed that the whole-cell current reduction was largely due to a significantly reduced open probability. Additionally, steady-state activation and inactivation were shifted to depolarized potentials by 15.9 and 5.1 mV, respectively. Systematic mutagenesis analysis of the spliced region provided evidence that a short amphiphilic region in the DII/DIII linker resembling an S4 voltage sensor of voltage-gated ion channels is an important determinant of Na(v)1.5 channel gating. Moreover, the present study identified novel short sequence motifs within this amphiphilic region that specifically affect the voltage dependence of steady state activation and inactivation and current amplitude of human Na(v)1.5. PMID- 16469735 TI - Identification of functional domains within the septation initiation network kinase, Cdc7. AB - The septation initiation network (SIN) serves to coordinate cytokinesis with mitotic exit in the fission yeast Schizosaccharomyces pombe. SIN components Spg1 and Cdc7 together play a central role in regulating the onset of septation and cytokinesis. Spg1, a Ras-like GTPase, localizes to the spindle pole bodies (SPBs) throughout the cell cycle. It is converted to its GTP-bound (active) state during mitosis, only to become inactivated at one SPB during anaphase and at both SPBs as cells exit mitosis. Cdc7 functions as an effector kinase for Spg1, binding to Spg1 in its GTP-bound state, and therefore is present at both SPBs during mitosis and asymmetrically at only one during anaphase. Interestingly, the kinase activity of Cdc7 does not vary across the cell cycle, suggesting the possibility that Cdc7 kinase activity is independent of Spg1 binding. Consistent with this, we found that Cdc7 associates with Spg1 only during mitosis. To learn more about the essential role of Cdc7 kinase in the SIN and its regulation, we undertook a structure/function analysis and identified independent functional domains within Cdc7. We found that a region adjacent to the kinase domain is responsible for Spg1 association and identified an overlapping but distinct SPB localization domain. In addition Cdc7 associates with itself and exists as a dimer in vivo. PMID- 16469734 TI - Regulation of epithelial Na+ channels (ENaC) by methylation: a novel methyltransferase stimulates ENaC activity. AB - Aldosterone acts to increase apical membrane permeability by activation of epithelial Na(+) channels (ENaC). We have previously shown that aldosterone activates ENaC early in the course of its action by stimulating the methylation of the beta subunit of this heteromeric channel in A6 cells. Aldosterone also stimulates the expression and methylation of k-ras in A6 cells. To determine whether aldosterone-stimulated methylations are seen in mammalian cells, we examined the effect of aldosterone on methylation and ras activation in a continuous line of cultured epithelial cells derived from mouse cortical collecting duct (CCD) and determined that beta mENaC is a substrate for methylation by an enzyme contained in CCD cells. Aldosterone stimulated protein base labile methylation in CCD cells. Aldosterone stimulated Na(+) transport in CCD cells within 1 h of addition and without an increase in cellular amount of any ENaC subunits over the first 4 h. Inhibition of methylation, using the inhibitor 3-deaza-adenosine, blocked the stimulation of Na(+) transport induced by aldosterone at early time points (1-4 h) without affecting cellular amounts of any ENaC subunits. In contrast to 3-deaza-adenosine (3-DZA), which inhibits all methylation reactions, specific inhibitors of small G-protein methylation or prenylation had no effect on the early aldosterone-induced current. Overexpression of isoprenylcysteine carboxylmethyltransferase (PCMTase), the enzyme that methylates ras, had little effect on basal transport but enhanced aldosterone-stimulated transport in A6 cells. Overexpression of PCMTase in CCD cells had no effect on either basal or aldosterone-stimulated transport. Moreover PCMTase had no effect on ENaC activity when co-expressed in Xenopus oocytes. Aldosterone had no effect on either message or protein levels of k-ras in CCD cells. Searching a mouse kidney library, we identified a methyltransferase that stimulates ENaC activity in Xenopus oocytes without affecting surface expression of ENaC. Our results demonstrate that aldosterone stimulates protein methylation in CCD cells, and this is required for expression of the early transport response. In CCD cells this effect is not mediated via methylation of ras, which is not induced by aldosterone in these cells, and the enzyme that methylates ras has no direct effect on ENaC activity. beta ENaC is a substrate for methylation in CCD cells. A novel methyltransferase that stimulates ENaC directly has been identified in CCD cells. PMID- 16469733 TI - Rho family Guanine nucleotide exchange factor Brx couples extracellular signals to the glucocorticoid signaling system. AB - Glucocorticoids regulate many crucial biologic functions through their cytoplasmic/nuclear glucocorticoid receptors (GR). Excess, deficiency, or alteration in tissue sensitivity to glucocorticoids has been associated with major causes of human morbidity and mortality. Brx, a cytoplasmic Rho family guanine nucleotide exchange factor, binds to and influences the activity of several nuclear hormone receptors. We examined the functional and molecular interactions between GR and Brx. The glucocorticoid sensitivity of lymphocytes obtained from mice haplo-insufficient for Brx was significantly decreased. Conversely, GR-mediated transcriptional activity of a glucocorticoid response element (GRE)-mediated glucocorticoid-responsive promoter was enhanced by Brx in a guanine nucleotide exchange factor domain-dependent fashion. Brx interacted with GR, forming a ternary complex with RhoA. In a chromatin immunoprecipitation assay, Brx and RhoA were co-precipitated with GREs only in the presence of ligand activated GR. Extracellularly administered lysophosphatidic acid, which activates its signaling cascade through a specific membrane GTP-binding protein (G-protein) coupled receptor in a G-protein alpha(13)-, Brx-, and RhoA-dependent fashion, enhanced GR transcriptional activity, whereas depletion of endogenous Brx attenuated this effect. These findings suggest that glucocorticoid signaling and, hence, the tissue sensitivity to glucocorticoids, may be coupled to extracellular signals via Brx and small G-proteins. Nuclear Brx might act as a local GRE-GR transcriptosome activator by mediating the effect of small G-proteins on glucocorticoid-regulated genes. PMID- 16469737 TI - New insights into the role of conserved, essential residues in the GTP binding/GTP hydrolytic cycle of large G proteins. AB - The GTP hydrolytic (GTPase) reaction terminates signaling by both large (heterotrimeric) and small (Ras-related) GTP-binding proteins (G proteins). Two residues that are necessary for GTPase activity are an arginine (often called the "arginine finger") found either in the Switch I domains of the alpha subunits of large G proteins or contributed by the GTPase-activating proteins of small G proteins, and a glutamine that is highly conserved in the Switch II domains of Galpha subunits and small G proteins. However, questions still exist regarding the mechanism of the GTPase reaction and the exact role played by the Switch II glutamine. Here, we have characterized the GTP binding and GTPase activities of mutants in which the essential arginine or glutamine residue has been changed within the background of a Galpha chimera (designated alpha(T)*), comprised mainly of the alpha subunit of retinal transducin (alpha(T)) and the Switch III region from the alpha subunit of G(i1). As expected, both the alpha(T)*(R174C) and alpha(T)*(Q200L) mutants exhibited severely compromised GTPase activity. Neither mutant was capable of responding to aluminum fluoride when monitoring changes in the fluorescence of Trp-207 in Switch II, although both stimulated effector activity in the absence of rhodopsin and Gbetagamma. Surprisingly, each mutant also showed some capability for being activated by rhodopsin and Gbetagamma to undergo GDP-[(35)S]GTPgammaS exchange. The ability of the mutants to couple to rhodopsin was not consistent with the assumption that they contained only bound GTP, prompting us to examine their nucleotide-bound states following their expression and purification from Escherichia coli. Indeed, both mutants contained bound GDP as well as GTP, with 35-45% of each mutant being isolated as GDP-P(i) complexes. Overall, these findings suggest that the R174C and Q200L mutations reveal Galpha subunit states that occur subsequent to GTP hydrolysis but are still capable of fully stimulating effector activity. PMID- 16469736 TI - The RACK1 homologue from Trypanosoma brucei is required for the onset and progression of cytokinesis. AB - The receptor for activated C kinase 1 (RACK1) is a conserved scaffold protein that helps regulate a range of cell activities including cell growth, shape, and protein translation. We report that a homologue of RACK1 is required for cytokinesis in pathogenic Trypanosoma brucei. The protein, referred to as TRACK, is comprised of WD repeat elements and can complement cpc2 null mutants of Schizosaccharomyces pombe. TRACK is expressed throughout the trypanosome life cycle and is distributed predominantly in a perinuclear region and the cytoplasm but not along the endoplasmic reticulum, mitochondrion, or cleavage furrow of dividing cells. When tetracycline-inducible RNA interference (RNAi) is used to deplete the cellular content of TRACK, the cells remain metabolically active, but growth is inhibited. In bloodstream forms, growth arrest is due to a delay in the onset of cytokinesis. By contrast, procyclic forms are able to initiate cytokinesis in the absence of TRACK but arrest midway through cell cleavage. The RNAi cells undergo multiple rounds of partial cytokinesis and accumulate nuclei and cytoplasmic extensions with attached flagella. The TRACK RNAi construct is also inducible within infected mice. Under these conditions parasites are eliminated from peripheral blood within 3 days post-infection. Taken as a whole, these data indicate that trypanosomes utilize a RACK1 homologue to regulate the final stages of mitosis. Moreover, disrupting the interaction between TRACK and its partners might be targeted in the design of novel therapies. PMID- 16469738 TI - Targeted disruption of tyrosylprotein sulfotransferase-2, an enzyme that catalyzes post-translational protein tyrosine O-sulfation, causes male infertility. AB - Tyrosine O-sulfation is a post-translational modification mediated by one of two Golgi tyrosylprotein sulfotransferases (TPST-1 and -2) expressed in all mammalian cells. Tyrosine sulfation plays an important role in the function of some known TPST substrates by enhancing protein-protein interactions. To explore the role of these enzymes in vivo and gain insight into other potential TPST substrates, TPST 2-deficient mice were generated by targeted disruption of the Tpst2 gene. Tpst2(+/-) mice appear normal and, when interbred, yield litters of normal size with a Mendelian distribution of the targeted mutation. Tpst2(-/-) mice have moderately delayed growth but appear healthy and attain normal body weight by 10 weeks of age. In contrast to Tpst1(-/-) males that have normal fertility, Tpst2( /-) males are infertile. Tpst2(-/-) sperm are normal in number, morphology, and motility in normal media and appear to capacitate and undergo acrosomal exocytosis normally. However, they are severely defective in their motility in viscous media and in their ability to fertilize zona pellucida-intact eggs. Adhesion of Tpst2(-/-) sperm to the egg plasma membrane is reduced compared with wild type sperm, but sperm-egg fusion is similar or even increased. These data strongly suggest that tyrosine sulfation of unidentified substrate(s) play a crucial role in these processes and document for the first time the critical importance of post-translational tyrosine sulfation in male fertility. PMID- 16469739 TI - Cysteine string protein monitors late steps in cystic fibrosis transmembrane conductance regulator biogenesis. AB - We examined the role of the cysteine string protein (Csp) in cystic fibrosis transmembrane conductance regulator (CFTR) biogenesis in relation to another J domain protein, Hdj-2, a recognized CFTR cochaperone. Increased expression of Csp produced a dose-dependent reduction in mature (band C) CFTR and an increase in immature (band B) CFTR. Exogenous expression of Hdj-2 also increased CFTR band B, but unlike Csp, Hdj-2 increased band C as well. The Csp-induced block of CFTR maturation required Hsp70, because a J-domain mutant (H43Q) that interferes with the ability of Csp to stimulate Hsp70 ATPase activity relieved the Csp-induced block of CFTR maturation. Nevertheless, Csp H43Q still increased immature CFTR. Csp-induced band B CFTR was found adjacent to the nucleus, co-localizing with calnexin, and it remained detergent-soluble. These data indicate that Csp did not block CFTR maturation by promoting the aggregation or degradation of immature CFTR. Csp knockdown by RNA interference produced a 5-fold increase in mature CFTR and augmented cAMP-stimulated CFTR currents. Thus, the production of mature CFTR is inversely related to the expression level of Csp. Both Csp and Hdj-2 associated with the CFTR R-domain in vitro, and Hdj-2 binding was displaced by Csp, suggesting common interaction sites. Combined expression of Csp and Hdj-2 mimicked the effect of Csp alone, a block of CFTR maturation. But together, Csp and Hdj-2 produced additive increases in CFTR band B, and this did not depend on their interactions with Hsp70, consistent with direct chaperone actions of these proteins. Like Hdj-2, Csp reduced the aggregation of NBD1 in vitro in the absence of Hsp70. Our data suggest that both Csp and Hdj-2 facilitate the biosynthesis of immature CFTR, acting as direct CFTR chaperones, but in addition, Csp is positioned later in the CFTR biogenesis cascade where it regulates the production of mature CFTR by limiting its exit from the endoplasmic reticulum. PMID- 16469740 TI - The latent membrane protein 1 of Epstein-Barr virus (EBV) primes EBV latency cells for type I interferon production. AB - Epstein-Barr virus (EBV) latency has been associated with a variety of human cancers. Latent membrane protein 1 (LMP-1) is one of the key viral proteins required for transformation of primary B cells in vitro and establishment of EBV latency. We have previously shown that LMP-1 induces the expression of several interferon (IFN)-stimulated genes and has antiviral effect (Zhang, J., Das, S. C., Kotalik, C., Pattnaik, A. K., and Zhang, L. (2004) J. Biol. Chem. 279, 46335 46342). In this report, a novel mechanism related to the antiviral effect of LMP 1 is identified. We show that EBV type III latency cells, in which LMP-1 is expressed, are primed to produce robust levels of endogenous IFNs upon infection of Sendai virus. The priming action is due to the expression of LMP-1 but not EBV nuclear antigen 2 (EBNA-2). The signaling events from the C-terminal activator regions of LMP-1 are essential to prime cells for high IFN production. LMP-1 mediated activation of NF-kappaB is apparently necessary and sufficient for LMP-1 mediated priming effect in DG75 cells, a human B cell line. IFN regulatory factor 7 (IRF-7) that can be activated by LMP-1 is also implicated in the priming action. Taken together, these data strongly suggest that LMP-1 may prime EBV latency cells for IFN production and that the antiviral property of LMP-1 may be an intrinsic part of EBV latency program, which may assist the establishment and/or maintenance of viral latency. PMID- 16469741 TI - A mutation in aminopeptidase N (CD13) isolated from a patient suffering from leukemia leads to an arrest in the endoplasmic reticulum. AB - Human aminopeptidase N (APN) is used as a routine marker for myelomonocytic cells in hematopoietic malignant disorders. Its gene and surface expressions are increased in cases of malignant transformation, inflammation, or T cell activation, whereas normal B and resting T cells lack detectable APN protein expression. In this study we elucidated the intracellular distribution, expression pattern, and enzymatic activity of a naturally occurring mutation in the coding region of the APN gene. At physiological temperatures the mutant protein is enzymatically inactive, persists as a mannose-rich polypeptide in the endoplasmic reticulum, and is ultimately degraded by an endoplasmic reticulum associated degradation pathway. It shows in part the distinct behavior of a temperature-sensitive mutant with a permissive temperature of 32 degrees C, leading to correct sorting of the Golgi compartment accompanied by the acquisition of proper glycosylation but without reaching the cell-surface membrane and without regaining its enzymatic activity. Because the patient bearing this mutation suffered from leukemia, possible links to the pathogenesis of leukemia are discussed. PMID- 16469742 TI - A chlorophyll a/b-binding protein homolog that is induced by iron deficiency is associated with enlarged photosystem I units in the eucaryotic alga Dunaliella salina. AB - Adaptation of the halotolerant alga Dunaliella salina to iron deprivation involves extensive changes of chloroplast morphology, photosynthetic activities, and induction of a major 45-kDa chloroplast protein termed Tidi. Partial amino acid sequencing of proteolytic peptides suggested that Tidi resembles chlorophyll a/b-binding proteins which compose light-harvesting antenna complexes (LHC) (Varsano, T., Kaftan, D., and Pick, U. (2003) J. Plant Nutr. 26, 2197-2210). Here we show that Tidi shares the highest amino acid sequence similarity with light harvesting I chlorophyll a/b-binding proteins from higher plants but has an extended proline-rich N-terminal domain. The accumulation of Tidi is reversed by iron supplementation, and its level is inversely correlated with photosystem I (PS-I) reaction center proteins. In native gel electrophoresis, Tidi co-migrates with enlarged PS-I-LHC-I super-complexes. Single particle electron microscopy analysis revealed that PS-I units from iron-deficient cells are larger (31 and 37 nm in diameter) than PS-I units from control cells (22 nm). The 77 K chlorophyll fluorescence emission spectra of isolated complexes suggest that the Tidi-LHC-I antenna are functionally coupled to the reaction centers of PS-I. These findings indicate that Tidi acts as an accessory antenna of PS-I. The enlargement of PS-I antenna in algae and in cyanobacteria under iron deprivation suggests a common limitation that requires rebalancing of the energy distribution between the two photosystems. PMID- 16469743 TI - NMR study of the electron transfer complex of plant ferredoxin and sulfite reductase: mapping the interaction sites of ferredoxin. AB - Plant ferredoxin serves as the physiological electron donor for sulfite reductase, which catalyzes the reduction of sulfite to sulfide. Ferredoxin and sulfite reductase form an electrostatically stabilized 1:1 complex for the intermolecular electron transfer. The protein-protein interaction between these proteins from maize leaves was analyzed by nuclear magnetic resonance spectroscopy. Chemical shift perturbation and cross-saturation experiments successfully mapped the location of two major interaction sites of ferredoxin: region 1 including Glu-29, Glu-30, and Asp-34 and region 2 including Glu-92, Glu 93, and Glu-94. The importance of these two acidic patches for interaction with sulfite reductase was confirmed by site-specific mutation of acidic ferredoxin residues in regions 1 and 2, separately and in combination, by which the ability of mutant ferredoxins to transfer electrons and bind to sulfite reductase was additively lowered. Taken together, this study gives a clear illustration of the molecular interaction between ferredoxin and sulfite reductase. We also present data showing that this interaction surface of ferredoxin significantly differs from that when ferredoxin-NADP(+) reductase is the interaction partner. PMID- 16469744 TI - Myocyte enhancer factors 2A and 2C induce dilated cardiomyopathy in transgenic mice. AB - Cardiac hypertrophy and dilation are mediated by neuroendocrine factors and/or mitogens as well as through internal stretch- and stress-sensitive signaling pathways, which in turn transduce alterations in cardiac gene expression through specific signaling pathways. The transcription factor family known as myocyte enhancer factor 2 (MEF2) has been implicated as a signal-responsive mediator of the cardiac transcriptional program. For example, known hypertrophic signaling pathways that utilize calcineurin, calmodulin-dependent protein kinase, and MAPKs can each affect MEF2 activity. Here we demonstrate that MEF2 transcription factors induced dilated cardiomyopathy and lengthening of myocytes. Specifically, multiple transgenic mouse lines with cardiac-specific overexpression of MEF2A or MEF2C presented with cardiomyopathy at base line or were predisposed to more fulminant disease following pressure overload stimulation. The cardiomyopathic response associated with MEF2A and MEF2C was not further altered by activated calcineurin, suggesting that MEF2 functions independently of calcineurin in this response. In cultured cardiomyocytes, MEF2A, MEF2C, and MEF2-VP16 overexpression induced sarcomeric disorganization and focal elongation. Mechanistically, MEF2A and MEF2C each programmed similar profiles of altered gene expression in the heart that included extracellular matrix remodeling, ion handling, and metabolic genes. Indeed, adenoviral transfection of cultured cardiomyocytes with MEF2A or of myocytes from the hearts of MEF2A transgenic adult mice showed reduced transient outward K(+) currents, consistent with the alterations in gene expression observed in transgenic mice and partially suggesting a proximal mechanism underlying MEF2-dependent cardiomyopathy. PMID- 16469745 TI - Activating transcription factor 3, a stress-inducible gene, suppresses Ras stimulated tumorigenesis. AB - ATF3 is a stress-inducible gene that encodes a member of the ATF/CREB family of transcription factors. Current literature indicates that ATF3 affects cell death and cell cycle progression. However, controversies exist, because it has been demonstrated to be a negative or positive regulator of these processes. We sought to study the roles of ATF3 in both cell death and cell cycle regulation in the same cell type using mouse fibroblasts. We show that ATF3 promotes apoptosis and cell cycle arrest. Fibroblasts deficient in ATF3 (ATF3(-/-)) were partially protected from UV-induced apoptosis, and fibroblasts ectopically expressing ATF3( /-) under the tet-off system exhibited features characteristic of apoptosis upon ATF3 induction. Furthermore, ATF3(-/-) fibroblasts transitioned from G(2) to S phase more efficiently than the ATF3(+/+) fibroblasts, suggesting a growth arrest role of ATF3. Consistent with the growth arrest and pro-apoptotic roles of ATF3, ATF3(-) fibroblasts upon Ras transformation exhibited higher growth rate, produced more colonies in soft agar, and formed larger tumor upon xenograft injection than the ATF3(+/+) counterparts. ATF3(-/-) cells, either with or without Ras transformation, had increased Rb phosphorylation and higher levels of various cyclins. Significantly, ATF3 bound to the cyclin D1 promoter as shown by chromatin immunoprecipitation (ChIP) assay and repressed its transcription by a transcription assay. Taken together, our results indicate that ATF3 promotes cell death and cell arrest, and suppresses Ras-mediated tumorigenesis. Potential explanations for the controversy about the roles of ATF3 in cell cycle and cell death are discussed. PMID- 16469746 TI - Molecular cloning and characterization of chick SPACRCAN. AB - MY-174, a monoclonal antibody that reacts with specific sialylated O-linked glycoconjugates of chick SPACR (sialoprotein associated with cones and rods), also recognizes another molecule of 300 kDa. Here, we verified that this 300-kDa molecule is chick SPACRCAN (sialoproteoglycan associated with cones and rods), another member of a novel interphotoreceptor matrix molecule family. Screening for chick SPACRCAN was carried out by plaque hybridization using a probe for chick SPACR. Specific polyclonal antibodies raised against chick SPACRCAN were used for the following experiments. To determine whether the 300-kDa molecule detected by MY-174 was identical to 300-kDa chick SPACRCAN, the migrations of these bands were examined after various glycosidase digestions. Furthermore, the expression levels were measured during retinal development and compared with those of chick SPACR. The results demonstrated that the 300-kDa molecule recognized by MY-174 was chick SPACRCAN, and we further identified it as a proteoglycan with chondroitin sulfate chains. SPACRCAN had heavily sialylated N- and O-linked glycoconjugates, and its MY-174 antigenicity was abolished by O glycanase treatment after neuraminidase treatment, as observed for chick SPACR. During retinal development, the mRNA and core protein expression levels, MY-174 antigenicity, and hyaluronan binding ability of SPACRCAN peaked around embryonic day 17 and then gradually decreased, whereas the corresponding expression levels of SPACR simply increased, but not its hyaluronan binding ability. The MY-174 reactivity of SPACRCAN in the adult retina was decreased compared with that in the newborn retina, whereas that of SPACR was increased. The decreased hyaluronan binding of SPACR was induced by an inhibitory effect of the excess of sialic acids in the adult stage. Thus, with similar core protein structures and specific sialylated glycoconjugates but distinct chondroitin sulfate chains, SPACRCAN and SPACR may have separate roles in the retina due to their differing expression profiles during development. PMID- 16469747 TI - Estrogen receptor-alpha binds p53 tumor suppressor protein directly and represses its function. AB - Estrogen receptor-alpha (ERalpha) promotes proliferation of breast cancer cells, whereas tumor suppressor protein p53 impedes proliferation of cells with genomic damage. Whether there is a direct link between these two antagonistic pathways has remained unclear. Here we report that ERalpha binds directly to p53 and represses its function. The activation function-2 (AF-2) domain of ERalpha and the C-terminal regulatory domain of p53 are necessary for the interaction. Knocking down p53 and ERalpha by small interfering RNA elicits opposite effects on p53-target gene expression and cell cycle progression. Remarkably, ionizing radiation that causes genomic damage disrupts the interaction between ERalpha and p53. Ionizing radiation together with ERalpha knock down results in additive effect on transcription of endogenous p53-target gene p21 (CDKN1) in human breast cancer cells. Our findings reveal a novel mechanism for regulating p53 and suggest that suppressing p53 function is an important component in the pro proliferative role of ERalpha. PMID- 16469748 TI - Triptolide, an inhibitor of the human heat shock response that enhances stress induced cell death. AB - Molecular chaperones, inducible by heat shock and a variety of other stresses, have critical roles in protein homeostasis, balancing cell stress with adaptation, survival, and cell death mechanisms. In transformed cells and tumors, chaperones are frequently overexpressed, with constitutive activation of the heat shock transcription factor HSF1 implicated in tumor formation. Here, we describe the activity of triptolide, a diterpene triepoxide from the plant Triptergium wilfordii, as an inhibitor of the human heat shock response. Triptolide treatment of human tissue culture cells prevented the inducible expression of heat shock genes, shown by suppression of an HSP70 promoter-reporter construct and by suppression of endogenous HSP70 gene expression. Upon examining the steps in the HSF1 activation pathway, we found that triptolide abrogates the transactivation function of HSF1 without interfering in the early events of trimer formation, hyperphosphorylation, and DNA binding. The ability of triptolide to inhibit the heat shock response renders these cells sensitive to stress-induced cell death, which may be of great relevance to cancer treatments. PMID- 16469749 TI - Individual Timp deficiencies differentially impact pro-MMP-2 activation. AB - Membrane-type matrix metalloproteinases (MT-MMPs) have emerged as key enzymes in tumor cell biology. The importance of MT1-MMP, in particular, is highlighted by its ability to activate pro-MMP-2 at the cell surface through the formation of a trimolecular complex comprised of MT1-MMP/tissue inhibitor of metalloproteinase-2 (TIMP-2)/pro-MMP-2. TIMPs 1-4 are physiological MMP inhibitors with distinct roles in the regulation of pro-MMP-2 processing. Here, we have shown that individual Timp deficiencies differentially affect MMP-2 processing using primary mouse embryonic fibroblasts (MEFs). Timp-3 deficiency accelerated pro-MMP-2 activation in response to both cytochalasin D and concanavalin A. Exogenous TIMP 2 and N-TIMP-3 inhibited this activation, whereas TIMP-3 containing matrix from wild-type MEFs did not rescue the enhanced MMP-2 activation in Timp-3(-/-) cells. Increased processing of MMP-2 did not arise from increased expression of MT1-MMP, MT2-MMP, or MT3-MMP or altered expression of TIMP-2 and MMP-2. To test whether increased MMP-2 processing in Timp-3(-/-) MEFs is dependent on TIMP-2, double deficient Timp-2(-/-)/-3(-/-) MEFs were used. In these double deficient cells, the cleavage of pro-MMP-2 to its intermediate form was substantially increased, but the subsequent cleavage of intermediate-MMP-2 to fully active form, although absent in Timp-2(-/-) MEFs, was detectable with combined Timp-2(-/-)/-3(-/-) deficiency. TIMP-4 associates with MMP-2 and MT1-MMP in a manner similar to TIMP 3, but its deletion had no effect on pro-MMP-2 processing. Thus, TIMP-3 provides an inherent regulation over the kinetics of pro-MMP-2 processing, serving at a level distinct from that of TIMP-2 and TIMP-4. PMID- 16469750 TI - Steady-state kinetics and inhibitory action of antitubercular phenothiazines on mycobacterium tuberculosis type-II NADH-menaquinone oxidoreductase (NDH-2). AB - Type-II NADH-menaquinone oxidoreductase (NDH-2) is an essential respiratory enzyme of the pathogenic bacterium Mycobacterium tuberculosis (Mtb) that plays a pivotal role in its growth. In the present study, we expressed and purified highly active Mtb NDH-2 using a Mycobacterium smegmatis expression system, and the steady-state kinetics and inhibitory actions of phenothiazines were characterized. Purified NDH-2 contains a non-covalently bound flavin adenine dinucleotide cofactor and oxidizes NADH with quinones but does not react with either NADPH or oxygen. Ubiquinone-2 (Q2) and decylubiquinone showed high electron-accepting activity, and the steady-state kinetics and the NADH-Q2 oxidoreductase reaction were found to operate by a ping-pong reaction mechanism. Phenothiazine analogues, trifluoperazine, Compound 1, and Compound 2 inhibit the NADH-Q2 reductase activity with IC50 = 12, 11, and 13 microm, respectively. Trifluoperazine inhibition is non-competitive for NADH, whereas the inhibition kinetics is found to be uncompetitive in terms of Q2. PMID- 16469751 TI - Mitochondrial function in Leydig cell steroidogenesis. AB - The first and rate-limiting step in the biosynthesis of steroid hormones is the transfer of cholesterol into mitochondria, which is facilitated by the steroidogenic acute regulatory (StAR) protein. Recent studies of Leydig cell function have focused on the molecular events controlling steroidogenesis; however, few studies have examined the importance of the mitochondria. The purpose of this investigation was to determine which aspects of mitochondrial function are necessary for Leydig cell steroidogenesis. MA-10 tumor Leydig cells were treated with 8-bromo-cAMP (cAMP) and site-specific mitochondrial disrupters, pro-oxidants, and their effects on progesterone synthesis, StAR expression, mitochondrial membrane potential (delta psi(m)) and ATP synthesis were determined. Dissipating delta psi(m) with CCCP inhibited progesterone synthesis, even in the presence of newly synthesized StAR protein. The electron transport inhibitor antimycin A significantly reduced cellular ATP, inhibited steroidogenesis, and reduced StAR protein expression. The F0/F1 ATPase inhibitor oligomycin reduced cellular ATP and inhibited progesterone synthesis and StAR protein expression, but had no effect on delta psi(m). Disruption of pH with nigericin significantly reduced progesterone production and StAR protein, but had minimal effects on delta psi(m). Sodium arsenite at low concentrations inhibited StAR protein but not mRNA expression and inhibited progesterone without disrupting delta psi(m). The mitochondrial Ca2+ inhibitor Ru360 also inhibited StAR protein expression. These results demonstrate that delta psi(m), ATP synthesis, delta pH and [Ca2+]mt are all required for steroid biosynthesis, and that mitochondria are sensitive to oxidative stress. These results suggest that mitochondria must be energized, polarized, and actively respiring to support Leydig cell steroidogenesis and alterations in the state of mitochondria may be involved in regulating steroid biosynthesis. PMID- 16469752 TI - Intra-arterial hepatic fotemustine for the treatment of liver metastases from uveal melanoma: experience in 101 patients. AB - BACKGROUND: Exclusive liver metastases occur in up to 40% of patients with uveal melanoma associated with a median survival of 2-7 months. Single agent response rates with commonly available chemotherapy are below 10%. We have investigated the use of fotemustine via direct intra-arterial hepatic (i.a.h.) administration in patients with uveal melanoma metastases. PATIENTS AND METHODS: A total of 101 patients from seven centers were treated with i.a.h. fotemustine, administered intra-arterially weekly for a 4-week induction period, and then as a maintenance treatment every 3 weeks until disease progression, unacceptable toxicity or patient refusal. RESULTS: A median of eight fotemustine infusions per patient were delivered (range 1-26). Catheter related complications occurred in 23% of patients; however, this required treatment discontinuation in only 10% of the patients. The overall response rate was 36% with a median overall survival of 15 months and a 2-year survival rate of 29%. LDH, time between diagnosis and treatment start and gender were significant predictors of survival. CONCLUSIONS: Locoregional treatment with fotemustine is well tolerated and seems to improve outcome of this poor prognosis patient population. Median survival rates are among the longest reported and one-third of the patients are still alive at 2 years. PMID- 16469753 TI - Multicenter phase III randomized trial of polychemotherapy (CVD regimen) versus the same chemotherapy (CT) plus subcutaneous interleukin-2 and interferon-alpha2b in metastatic melanoma. AB - BACKGROUND: The addition of cytokines to chemotherapy (CT) has obtained encouraging but contradictory results in metastatic melanoma. In this phase III trial, we compared the effects of CT [cisplatin, vindesine and dacarbazine (CVD)] with those of concurrent biochemotherapy (bioCT) consisting of CVD plus interleukin-2 and interferon-alpha2b. PATIENTS AND METHODS: A total of 151 untreated metastatic melanoma patients were randomized, 75 on arm A (cisplatin 30 mg/m2 on days 1-3, vindesine 2.5 mg/m2 on day 1 and dacarbazine 250 mg/m2 on days 1-3), and 76 on arm B (same CVD scheme plus interferon-alpha2b on days 1-5 and interleukin-2 on days 1-5 and 8-15, both administered subcutaneously), either recycled every 3 weeks. Response was assessed every two cycles. RESULTS: Ten percent of the patients were alive at a median of 52 months from start of therapy. We observed a response rate (RR) of 21% on arm A versus 33% on arm B; three patients (4%) given bioCT had complete responses (CRs). Median time to progression (TTP) was identical; median overall survival (OS) time was 12 months on arm A and 11 months on arm B. CONCLUSIONS: BioCT is not better than CT alone; the trend in favor of the bioCT in terms of RR did not translate into better TTP or OS. Therefore, bioCT cannot be recommended as standard first-line therapy for metastatic melanoma. PMID- 16469754 TI - Biphasic metaplastic sarcomatoid carcinoma of the breast. AB - BACKGROUND: Breast biphasic metaplastic sarcomatoid carcinoma (MSC) is rare and aggressive. We analyzed 100 patients treated at M. D. Anderson Cancer Center (MDACC) with 213 MSC and 98 carcinosarcoma patients identified through the Surveillance, Epidemiology and End-Results (SEER) database to describe clinical and pathologic characteristics. PATIENTS AND METHODS: We searched the MDACC (1985 2001) and SEER databases (1988-2001) for breast MSC and carcinosarcoma patients. RESULTS: We identified 100 MDACC MSC patients: 66% had node-negative disease and 6% distant metastases at presentation. Median recurrence-free survival (RFS) of 94 patients with stages I-III disease was 74 months (range 3-74), with 52% 5-year RFS [95% confidence interval (CI) 0.42-0.63]. Median overall survival in these patients was not reached, with 64% 5-year survival (95% CI 0.54-0.75). The initial stage of the tumor, but not use of adjuvant chemo- or radiotherapy, had a strong association with outcome. The pathologic complete response rate to neoadjuvant chemotherapy was 10%. Median survival from the time of recurrent disease was 14 months (range 1-55). Tumors were usually hormone receptor- and HER2/neu-negative. SEER data were consistent with MDACC findings. CONCLUSIONS: Breast MSC and carcinosarcoma are aggressive, treatment-refractory tumors with shared clinical features and outcome similar to poorly differentiated receptor negative adenocarcinomas. New therapeutic agents are needed. PMID- 16469756 TI - Famine, Turks, and plague: impressions from Graz. PMID- 16469755 TI - The activity of methylated and non-methylated selenium species in lymphoma cell lines and primary tumours. AB - BACKGROUND: Diffuse large B-cell lymphoma patients with low serum selenium concentration at presentation have a lower response rate and overall survival than patients with higher serum selenium. The co-administration of selenium with conventional chemotherapy may be useful in these patients. PATIENTS AND METHODS: We investigated the activity of two selenium species, methylseleninic acid (MSA) and selenodiglutathione (SDG) in a panel of human lymphoma cell lines and in a primary lymphoma culture system. RESULTS: Both compounds demonstrated cytostatic and cytotoxic activity with EC(50) values in the range 1.0-10.2 microM. Cell death was associated with an increase in the sub-G1 (apoptotic) fraction by flow cytometry and was not preceded by any obvious cell cycle arrest. SDG, but not MSA, resulted in marked increases in intracellular ROS, particularly in CRL2261 and SUD4 cells in which the cytotoxic activity of SDG was partly, or completely, inhibited by n-acetyl cysteine, suggesting a dependence on ROS for activity in some cells. Both MSA and SDG showed a concentration dependent reduction in percentage viability after a 2-day exposure in primary lymphoma cultures, with EC(50) values in the range 39-300 microM and 9-28 microM, respectively. CONCLUSION: The selenium compounds MSA and SDG induce cell death in lymphoma cell lines and primary lymphoma cultures, which with SDG may be partly attributable to the generation of ROS. PMID- 16469757 TI - Local health systems in 21st century: who cares?-An exploratory study on health system governance in Amsterdam. AB - BACKGROUND: There is a growing awareness that there should be a public health perspective to health system governance. Its intrinsic population health orientation provides the ultimate ground for determining the health needs and governing collaborative care arrangements within which these needs can be met. Notwithstanding differences across countries, population health concerns are not central to European health reforms. Governments currently withdraw leaving governance roles to care providers and/or financiers. Thereby, incentives that trigger the uptake of a public health perspective are often ignored. METHODS: In this study we addressed this issue in the city of Amsterdam. Using a qualitative study design, we explored whether there is a public health perspective to the governance practices of the municipality and the major sickness fund in Amsterdam. And if so, what the scope of this perspective is. And if not, why not. RESULTS: Findings indicate that the municipality has a public health perspective to local health system governance, but its scope is limited. The municipality facilitates rather than governs health care provision in Amsterdam. Furthermore, the sickness fund runs major financial risks when adapting a public health perspective. It covers an insured population that partly overlaps the Amsterdam population. Returns on investments in population health are therefore uncertain, as competitors would also profit from the sickness fund's investments. CONCLUSION: The local health system in Amsterdam is not consistently aligned to the health needs of the Amsterdam population. The Amsterdam case is not unique and general consequences for local health system governance are discussed. PMID- 16469758 TI - Breastfeeding beliefs and practices among migrant mothers in slums of Diyarbakir, Turkey, 2001. AB - BACKGROUND: A qualitative investigation and a population survey were conducted to explore the breastfeeding beliefs and practices of mothers who were forced to migrate from their original villages and were currently living in the slums of Diyarbakir in Turkey. METHODS: Qualitative data collection on breastfeeding beliefs was conducted using in-depth interviews. In-depth interviews were tape recorded. Quantitative data on breastfeeding practices were collected using a structured questionnaire. RESULTS: Mothers generally have a positive attitude towards breastfeeding, but colostrum is usually perceived negatively. No woman was found to feed her infant exclusively by breastfeeding. Only 9.9% of mothers initiated breastfeeding within the first hour of birth. Forty per cent of mothers started solid foods before 4 months. Mother's education appeared as a significant factor influencing the introduction of colostrum to the newborn. Mothers with lower education generally believed that the colostrum should not be fed to the infant and that a pregnant woman's milk is unhealthy for the baby. There was also a belief that 'working under the sun' decreased the quality of milk of a mother. CONCLUSION: Cultural beliefs have a significant influence on breastfeeding practices. Some of these practices are potentially harmful to newborns. Health education programmes should address these beliefs and practices in culture sensitive ways. PMID- 16469759 TI - The German cervical cancer screening model: development and validation of a decision-analytic model for cervical cancer screening in Germany. AB - BACKGROUND: We sought to develop and validate a decision-analytic model for the natural history of cervical cancer for the German health care context and to apply it to cervical cancer screening. METHODS: We developed a Markov model for the natural history of cervical cancer and cervical cancer screening in the German health care context. The model reflects current German practice standards for screening, diagnostic follow-up and treatment regarding cervical cancer and its precursors. Data for disease progression and cervical cancer survival were obtained from the literature and German cancer registries. Accuracy of Papanicolaou (Pap) testing was based on meta-analyses. We performed internal and external model validation using observed epidemiological data for unscreened women from different German cancer registries. The model predicts life expectancy, incidence of detected cervical cancer cases, lifetime cervical cancer risks and mortality. RESULTS: The model predicted a lifetime cervical cancer risk of 3.0% and a lifetime cervical cancer mortality of 1.0%, with a peak cancer incidence of 84/100,000 at age 51 years. These results were similar to observed data from German cancer registries, German literature data and results from other international models. Based on our model, annual Pap screening could prevent 98.7% of diagnosed cancer cases and 99.6% of deaths due to cervical cancer in women completely adherent to screening and compliant to treatment. Extending the screening interval from 1 year to 2, 3 or 5 years resulted in reduced screening effectiveness. CONCLUSIONS: This model provides a tool for evaluating the long term effectiveness of different cervical cancer screening tests and strategies. PMID- 16469760 TI - Identification of novel proteases and immunomodulators in the secretions of schistosome cercariae that facilitate host entry. AB - Schistosomiasis, caused by parasitic helminths, remains a serious human disease in the tropics. Cercariae of Schistosoma mansoni infect their hosts by direct skin penetration, aided by secretions from acetabular and head glands. Both proteolytic and immunomodulatory properties have been ascribed to the released material, but to date only five isoforms of elastase and one putative anti inflammatory protein (Sm16) have been cloned. We analyzed secretions from mechanically transformed cercariae by two-dimensional electrophoresis. An average gel image was created and compared with a separation of soluble larval extract, revealing a less complex spot pattern in the secretions with 60% of the spots matched to the larval extract. Subsequent tandem mass spectrometric analysis identified 48 spots from the released material, representing approximately 80% of its normalized volume. Twenty-nine of these are likely to originate in the vesicles, and 18 are likely to originate in the cytosol of the glands (the latter class being present due to holocrine secretion); one is unknown. The vesicular proteins were significantly more enriched than the cytosolic proteins in the released material when compared with the larval extract. A novel metalloproteinase (termed SmPepM8) was the second most abundant constituent after three isoforms of cercarial elastase. In addition, a dipeptidyl peptidase IV (SmDPP IV) was discovered but in much smaller quantity. A new serine protease inhibitor (SmSerp_c) was also prominent. Along with Sm16, four potential immunomodulators were identified, three with similarity to venom allergens (SmSCP_a, _b, and _c) and one with homology to the potassium channel blockers in scorpion venom (SmKK7). Interrogation of the expressed sequence tag database found transcripts encoding the majority of vesicular proteins present solely in the intramolluscan stages of the life cycle. Distinct patterns of radiolabel incorporation suggested three separate origins for the vesicular proteins. All the novel constituents merit investigation as vaccine candidates, and the potential immunomodulators merit investigation as therapeutic agents. PMID- 16469761 TI - Accessing research participants in schools: a case study of a UK adolescent sexual health survey. AB - While methods and results of school-based studies have been reported widely in the literature, little published information exists on the practical aspects of recruiting schools and students into a study. This paper reflects on the experiences of a UK-based sexual health survey among 3007 students aged 15-18 years. The survey explored beliefs, attitudes and behaviours in relation to sexual health. This case study highlights significant aspects of planning and conducting successful large-scale research in schools, focusing on the process of conducting the research rather than outcomes. As such, the paper will benefit those intending to sample a school-based population. The key features of effective and feasible research in schools are outlined in four areas: (i) adopting suitable research tools, (ii) selecting and contacting schools, (iii) selecting students within schools and (iv) the importance of fieldworkers. On site and post-data collection feedback from teachers are incorporated into the discussion of good practice in partnership working with schools and students in research. We conclude by discussing fieldwork experiences and outlining key recommendations for researchers across disciplines engaging in school-based studies. PMID- 16469762 TI - Ticlopidine-induced lupus with renal involvement. PMID- 16469763 TI - Comparison of peritoneal dialysis and haemodialysis after renal transplant failure. AB - BACKGROUND: A growing number of patients are returning to dialysis after renal transplant failure. The aim of this study is to determine whether peritoneal dialysis (PD) is a safe and good treatment option for these patients. METHODS: All patients returning to PD or haemodialysis (HD) after renal transplant failure before 1 October 2002 at the University Hospital Gasthuisberg, Leuven, Belgium, were evaluated. Data were collected until death, retransplantation (reTx), transfer to HD or PD or until 1 January 2003. RESULTS: Twenty-one patients starting PD (PDpostTx-group) and 39 patients starting HD (HDpostTx-group) after renal transplant failure were included in the study. There were no significant differences in age, sex, serum albumin- and CRP-levels at baseline. The total time on renal replacement therapy at transplant failure and time to transplant failure did not differ between the two groups either. Furthermore, the baseline comorbidity was similar in both groups. During follow-up, the outcome did not differ significantly between the two groups. However, there was a tendency towards higher patient survival and reTx tended to be more frequent in the PDpostTx-group. Moreover, patients in the HDpostTx-group tended to accrue more new comorbidity. The incidence of peritonitis and the evolution of dialysis adequacy (renal and peritoneal Kt/V and creatinine clearances) with time in the PDpostTx-group was similar to that seen in our centre's PD patients who had never undergone transplantation before. CONCLUSIONS: This study suggests that the outcome in patients starting PD after renal transplant failure is at least as good as the outcome in those starting HD. Although these observational findings warrant further confirmation, PD therefore can be regarded as a safe and good treatment option for patients returning to dialysis after renal transplant failure. PMID- 16469764 TI - Quality assurance of neuroradiology in a District General Hospital. AB - BACKGROUND: An increasing number of neurologists in District General Hospitals (DGHs) rely on local neuroimaging reports from general radiologists. AIM: To determine the level of disagreement between general radiologists and neuroradiologists in reporting neuroimaging from patients referred to a neurologist. DESIGN: Prospective observational study. METHODS: We studied 232 patients referred for a neuroradiologist's report on neuroimaging over a 17-month period. Pre-planned comparisons included primary and secondary report findings, length of report and suggestions for additional investigations. RESULTS: Of the 593 patients assessed during the study period, a neuroradiologist's report was sought for 232 (39%): 119 men, 113 women, mean age 46.1 (SD 17.6) years. Primary findings differed in 37 patients (15.9%) (95%CI 11.5-21.3). Reports from neuroradiologists changed subsequent management in 31 (13.4%) (95%CI 9.3-18.4). Differences in secondary findings occurred in 52 (22.4%) (95%CI 17.2-28.3), and differences in either primary or secondary outcomes in 77 (33.2%) (95%CI 27.2 39.6). The level of disagreement in primary findings was as frequent among patients investigated with magnetic resonance imaging as among computerized tomogram-only patients (p = 0.13). Neuroradiologists recommended additional investigations for 24 patients (10.3%) (95%CI 6.7-15.0) and provided longer reports than general radiologists (p < 0.001). DISCUSSION: Neuroimaging reports of some patients differ substantially between general radiologists and neuroradiologists. Optimal management of neurological patients in DGHs may require timely access to neuroradiologists. PMID- 16469765 TI - Unusual causes of hypokalaemia and paralysis. AB - We demonstrate how the application of physiological principles may help to identify unusual causes of a very low plasma potassium (K+) concentration (P(K)) and paralysis. In the two patients described, the short time course of the illness suggested that there was an acute shift of K+ into cells. The combination of a low rate of excretion of K+, the absence of a metabolic acid-base disorder, and the fact that the clinical findings occurred very soon after a large intake of carbohydrate supported this impression. Surprisingly, the P(K) remained low for many hours after these stimuli to shift K+ into cells had abated. The missing link in this story was eventually provided by the attending medical team with the help of their mentor, Professor McCance. PMID- 16469766 TI - Mash1 is required for generic and subtype differentiation of hypothalamic neuroendocrine cells. AB - The neuroendocrine hypothalamus regulates a number of critical biological processes and underlies a range of diseases from growth failure to obesity. Although the elucidation of hypothalamic function has progressed well, knowledge of hypothalamic development is poor. In particular, little is known about the processes underlying the neurogenesis and specification of neurons of the ventral nuclei, the arcuate and ventromedial nuclei. The proneural gene Mash1 is expressed throughout the basal retrochiasmatic neuroepithelium and loss of Mash1 results in hypoplasia of both the arcuate and ventromedial nuclei. These defects are due to a failure of neurogenesis and apoptosis, a defect that can be rescued by ectopic Ngn2 under the control of the Mash1 promoter. In addition to its role in neurogenesis, analysis of Mash1(-/-), Mash1(+/-), Mash1(KINgn2/KINgn2), and Mash1(KINgn2/+) mice demonstrates that Mash1 is specifically required for Gsh1 expression and subsequent GHRH expression, positively regulates SF1 expression, and suppresses both tyrosine hydroxylase (TH) and neuropeptide Y (NPY) expression. Although Mash1 is not required for propiomelanocortin (POMC) expression, it is required for normal development of POMC(+) neurons. These data demonstrate that Mash1 is both required for the generation of ventral neuroendocrine neurons as well as playing a central role in subtype specification of these neurons. PMID- 16469767 TI - Socs2 and elf5 mediate prolactin-induced mammary gland development. AB - The proliferative phase of mammary alveolar morphogenesis is initiated during early pregnancy by rising levels of serum prolactin and progesterone, establishing a program of gene expression that is ultimately responsible for the development of the lobuloalveoli and the onset of lactation. To explore this largely unknown genetic program, we constructed transcript profiles derived from transplanted mammary glands formed by recombination of prolactin receptor (Prlr) knockout or wild-type mammary epithelium with wild-type mammary stroma. Comparison with profiles derived from prolactin-treated Scp2 mammary epithelial cells produced a small set of commonly prolactin-regulated genes that included the negative regulator of cytokine signaling, Socs2 (suppressor of cytokine signaling 2), and the ets transcription factor, E74-like factor 5 (Elf5). Homozygous null mutation of Socs2 rescued the failure of lactation and reduction of mammary signal transducer and activator of transcription 5 phosphorylation that characterizes Prlr heterozygous mice, demonstrating that mammary Socs2 is a key regulator of the prolactin-signaling pathway. Reexpression of Elf5 in Prlr nullizygous mammary epithelium restored lobuloalveolar development and milk production, demonstrating that Elf5 is a transcription factor capable of substituting for prolactin signaling. Thus, Socs2 and Elf5 are key members of the set of prolactin-regulated genes that mediate prolactin-driven mammary development. PMID- 16469768 TI - Sex-dependent liver gene expression is extensive and largely dependent upon signal transducer and activator of transcription 5b (STAT5b): STAT5b-dependent activation of male genes and repression of female genes revealed by microarray analysis. AB - Sexual dimorphism in mammalian liver contributes to sex differences in physiology, homeostasis, and steroid and foreign compound metabolism. Many sex dependent liver genes are regulated by sex differences in pituitary GH secretion, with the transcription factor, signal transducer and activator of transcription (STAT5b), proposed to mediate signaling by the pulsatile, male plasma GH profile. Presently, a large-scale gene expression study was conducted using male and female mice, wild type and Stat5b inactivated, to characterize sex differences in liver gene expression and their dependence on STAT5b. The relative abundance of individual liver RNAs was determined for each sex-genotype combination by competitive hybridization to 23,574-feature oligonucleotide microarrays. Significant sex differences in hepatic expression were seen for 1603 mouse genes. Of 850 genes showing higher expression in males, 767 (90%) were down-regulated in STAT5b-deficient males. Moreover, of 753 genes showing female-predominant expression, 461 (61%) were up-regulated in STAT5b-deficient males. In contrast, approximately 90% of the sex-dependent genes were unaffected by STAT5b deficiency in females. Thus: 1) STAT5b is essential for sex-dependent liver gene expression, a characteristic of approximately 1600 mouse genes (4% of the genome); 2) male predominant liver gene expression requires STAT5b, or STAT5b-dependent factors, which act in a positive manner; and 3) many female-predominant liver genes are repressed in males in a STAT5b-dependent manner. Several of the STAT5b-dependent male genes encode transcriptional repressors; these may include direct STAT5b targets that repress female-predominant genes in male liver. Several female predominant repressors are elevated in STAT5b-deficient males; these may contribute to the major loss of male gene expression seen in the absence of STAT5b. PMID- 16469769 TI - P190-B Rho GTPase-activating protein overexpression disrupts ductal morphogenesis and induces hyperplastic lesions in the developing mammary gland. AB - p190-B Rho GTPase activating protein is essential for mammary gland development because p190-B deficiency prevents ductal morphogenesis. To investigate the role of p190-B during distinct stages of mammary gland development, tetracycline regulatable p190-B-overexpressing mice were generated. Short-term induction of p190-B in the developing mammary gland results in abnormal terminal end buds (TEBs) that exhibit aberrant budding off the neck, histological anomalies, and a markedly thickened stroma. Overexpression of p190-B throughout postnatal development results in increased branching, delayed ductal elongation, and disorganization of the ductal tree. Interestingly, overexpression of p190-B during pregnancy results in hyperplastic lesions. Several cellular and molecular alterations detected within the aberrant TEBs may contribute to these phenotypes. Signaling through the IGF pathway is altered, and the myoepithelial cell layer is discontinuous at sites of aberrant budding. An increase in collagen and extensive infiltration of macrophages, which have recently been implicated in branching morphogenesis, is observed in the stroma surrounding the p190-B-overexpressing TEBs. We propose that the stromal response, disruption of the myoepithelial layer, and alterations in IGF signaling in the p190-B-overexpressing mice impact the TEB architecture, leading to disorganization and increased branching of the ductal tree. Moreover, we suggest that alterations in tissue architecture and the adjacent stroma as a consequence of p190-B overexpression during pregnancy leads to loss of growth control and the formation of hyperplasia. These data demonstrate that precise control of p190-B Rho GTPase-activating protein activity is critical for normal branching morphogenesis during mammary gland development. PMID- 16469770 TI - Involvement of p38 mitogen-activated protein kinase and inducible nitric oxide synthase in apoptotic signaling of murine and human male germ cells after hormone deprivation. AB - This study investigates the role of p38 MAPK, inducible nitric oxide synthase (iNOS), and the intrinsic pathway signaling in male germ cell death in rats after hormonal deprivation by a potent GnRH antagonist treatment. Germ cell apoptosis, involving exclusively middle (VII-VIII) stages, was activated by d 5 after GnRH antagonist treatment. Initiation of germ cell apoptosis was preceded by p38 MAPK activation and induction of iNOS. p38 MAPK activation and iNOS induction were further accompanied by a marked perturbation of the BAX/BCL-2 rheostat, cytochrome c, and DIABLO release from mitochondria, caspase activation, and poly(ADP-ribose) polymerase cleavage. Concomitant administration of aminoguanidine, a selective iNOS inhibitor, significantly prevented hormone deprivation-induced germ cell apoptosis. Inhibitors of iNOS or p38 MAPK were also effective in preventing human male germ cell apoptosis induced by hormone-free culture conditions. Together, these results establish a new signal transduction pathway involving p38 MAPK and iNOS that, through activation of the intrinsic pathway signaling, promotes male germ cell death in response to a lack of hormonal stimulation across species. PMID- 16469771 TI - In vitro evidence that upstream of N-ras participates in the regulation of parathyroid hormone messenger ribonucleic acid stability. AB - Calcium and phosphate regulate PTH gene expression posttranscriptionally through the binding of trans-acting factors to a defined cis-acting instability element in the PTH mRNA 3'-untranslated region (UTR). We have previously defined AU-rich binding factor 1 as a PTH mRNA binding and stabilizing protein. We have now identified, by affinity chromatography, Upstream of N-ras (Unr) as another PTH mRNA 3'-UTR binding protein. Recombinant Unr bound the PTH 3'-UTR transcript, and supershift experiments with antibodies to Unr showed that Unr is part of the parathyroid RNA binding complex. Finally, because there is no parathyroid cell line, the functionality of Unr in regulating PTH mRNA levels was demonstrated in cotransfection experiments in heterologous human embryonic kidney 293 cells. Depletion of Unr by small interfering RNA decreased simian virus 40-driven PTH gene expression in human embryonic kidney 293 cells transiently cotransfected with the human PTH gene. Overexpression of Unr increased the rat full-length PTH mRNA levels but not a PTH mRNA lacking the terminal 60-nucleotide cis-acting protein binding region. Unr also stabilized a chimeric GH reporter mRNA that contained the rat PTH 63-nucleotide cis-acting element but not a truncated PTH element. Therefore, Unr binds to the PTH cis element and increases PTH mRNA levels, as does AU-rich binding factor 1. Our results suggest that Unr, together with the other proteins in the RNA binding complex, determines PTH mRNA stability. PMID- 16469772 TI - Activation function 1 of glucocorticoid receptor binds TATA-binding protein in vitro and in vivo. AB - The mechanism through which the glucocorticoid receptor (GR) stimulates transcription is still unclear, although it is clear that the GR affects assembly of the transcriptional machinery. The binding of the TATA-binding protein (TBP) to the TATA-box is accepted as essential in this process. It is known that the GR can interact in vitro with TBP, but the direct interaction of TBP with GR has not been previously characterized quantitatively and has not been appreciated as an important step in assembling the transcriptional complex. Herein, we demonstrate that the TBP-GR interaction is functionally significant by characterizing the association of TBP and GR in vitro by a combination of techniques and confirming the role of this interaction in vivo. Combined analysis, using native gel electrophoresis, sedimentation equilibrium, and isothermal microcalorimetry titrations, characterize the stoichiometry, affinity, and thermodynamics of the TBP-GR interaction. TBP binds recombinant GR activation function 1 (AF1) with a 1:2 stoichiometry and a dissociation constant in the nanomolar range. In vivo fluorescence resonance energy transfer experiments, using fluorescently labeled TBP and various GR constructs, transiently transfected into CV-1 cells, show GR TBP interactions, dependent on AF1. AF1-deletion variants showed fluorescence resonance energy transfer efficiencies on the level of coexpressed cyan fluorescent protein and yellow fluorescent protein, indicating that the interaction is dependent on AF1 domain. To demonstrate the functional role of the in vivo GR-TBP interaction, increased amounts of TBP expressed in vivo stimulated expression of GR-driven reporters and endogenous genes, and the effect was also specifically dependent on AF1. PMID- 16469773 TI - Gastric inhibitory polypeptide as an endogenous factor promoting new bone formation after food ingestion. AB - Calcium plays a fundamental role as second messenger in intracellular signaling and bone serves as the body's calcium reserve to tightly maintain blood calcium levels. Calcium in ingested meal is the main supply and inadequate calcium intake causes osteoporosis and bone fracture. Here, we describe a novel mechanism of how ingested calcium is deposited on bone. Meal ingestion elicits secretion of the gut hormone gastric inhibitory polypeptide (GIP) from endocrine K cells in the duodenum. Bone histomorphometrical analyses revealed that bone formation parameters in the mice lacking GIP receptor (GIPR(-/-)) were significantly lower than those of wild-type (GIPR(+/+)) mice, and that the number of osteoclasts, especially multinuclear osteoclasts, was significantly increased in GIPR(-/-) mice, indicating that GIPR(-/-) mice have high-turnover osteoporosis. In vitro examination showed the percentage of osteoblastic cells undergoing apoptosis to be significantly decreased in the presence of GIP. Because GIPR(-/-) mice exhibited an increased plasma calcium concentration after meal ingestion, GIP directly links calcium contained in meal to calcium deposition on bone. PMID- 16469774 TI - A novel activating mutation in the RET tyrosine kinase domain mediates neoplastic transformation. AB - We report the finding of a novel missense mutation at codon 833 in the tyrosine kinase of the RET proto-oncogene in a patient with a carcinoma of the thyroid. In vitro experiments demonstrate that the R833C mutation induces transformed foci only when present in the long 3' splice isoform and, in keeping with a model in which the receptor has to dimerize to be completely activated, glial cell line derived neurotrophic factor stimulation leads the RET(R833C) receptor to a higher level of activation. Tyrosine kinase assays show that the RET(R833C) long isoform has weak intrinsic kinase activity and phosphorylation of an exogenous substrate is not elevated even in the presence of glial cell line-derived neurotrophic factor. Furthermore, the R833C mutation is capable of sustaining the transformed phenotype in vivo but does not confer upon the transformed cells the ability to degrade the basement membrane in a manner analogous to metastasis. Our functional characterization of the R833C substitution suggests that, like the V804M and S891A mutations, this tyrosine kinase mutation confers a weak activating potential upon RET. This is the first report demonstrating that the introduction of an intracellular cysteine can activate RET. However, this does not occur via dimerization in a manner analogous to the extracellular cysteine mutants. PMID- 16469776 TI - Understanding efficiency of human muscular movement exemplifies integrative and translational physiology. PMID- 16469775 TI - Fast IPSCs in rat thalamic reticular nucleus require the GABAA receptor beta1 subunit. AB - Synchrony within the thalamocortical system is regulated in part by intranuclear synaptic inhibition within the reticular nucleus (RTN). Inhibitory postsynaptic currents (IPSCs) in RTN neurons are largely characterized by slow decay kinetics that result in powerful and prolonged suppression of spikes. Here we show that some individual RTN neurons are characterized by highly variable mixtures of fast, slow and mixed IPSCs. Heterogeneity arose largely through differences in the contribution of an initial decay component (tau(D) approximately 10 ms) which was insensitive to loreclezole, suggesting involvement of the GABA(A) receptor beta(1) subunit. Single-cell RT-PCR revealed the presence of beta(1) subunit mRNA only in those neurons whose IPSCs were dominated by a rapid and prominent initial decay phase. These data show that brief, beta(1)-dependent, loreclezole insensitive IPSCs are present in a subpopulation of RTN neurons, and suggest that striking differences in IPSC heterogeneity within single neurons can result from of the presence or absence of a single GABA(A) receptor subunit. PMID- 16469777 TI - Calcium signalling in muscle: a milestone for modulation studies. PMID- 16469778 TI - Fetal exposure to excess glucocorticoid is unlikely to explain the effects of periconceptional undernutrition in sheep. AB - Periconceptional undernutrition alters fetal growth, metabolism and endocrinology in late gestation. The underlying mechanisms remain uncertain, but fetal exposure to excess maternal glucocorticoids has been hypothesized. We investigated the effects of periconceptional undernutrition on maternal hypothalamic-pituitary adrenal axis function and placental 11beta-hydroxysteroid dehydrogenase type 2 (11betaHSD2) activity. Ewes received maintenance feed (N, n= 20) or decreased feed from -60 to +30 days from mating to achieve 15% weight loss after an initial 2-day fast (UN, n= 21). Baseline plasma samples and arginine vasopressin (AVP) corticotrophin-releasing hormone (CRH) challenges were performed on days -61, 57, -29, -1, +29, 33, and 49 from mating (day 0). Maternal adrenal and placental tissue was collected at 50 days. Baseline plasma levels of adrenocorticotrophic hormone (ACTH) and cortisol decreased in the UN group (P < 0.0001). ACTH response to AVP-CRH was greater in UN ewes during undernutrition (P= 0.03) returning to normal levels after refeeding. Cortisol response to AVP-CRH was greater in UN ewes after the initial 2-day fast, but thereafter decreased and was lower in UN ewes from mating until the end of the experiment (P= 0.007). ACTH receptor, StAR and p450c17 mRNA levels were down-regulated in adrenal tissue from UN ewes. Placental 11betaHSD2 activity was lower in UN than N ewes at 50 days (P= 0.014). Moderate periconceptional undernutrition results in decreased maternal plasma cortisol concentrations during undernutrition and after refeeding, and adrenal resistance to ACTH for at least 20 days after refeeding. Fetal exposure to excess maternal cortisol is unlikely during the period of undernutrition, but could occur later in gestation if maternal plasma cortisol levels return to normal while placental 11betaHSD2 activity remains low. PMID- 16469779 TI - Development of baroreflex function and hind limb vascular reactivity in the horse fetus. AB - This study investigated, in vivo, the mechanisms underlying the development of cardiovascular function in the horse fetus, with particular relevance to baroreflex function and hind limb vascular arterial reactivity to constrictor agonists. Under general anaesthesia, vascular catheters were inserted and a Transonic flow probe was implanted around one of the metatarsal arteries of 13 horse fetuses, either at 0.6 of gestation (n= 6) or at 0.9 of gestation (n= 7, term approximately 335 days). At least 5 days after surgery, pressor, vasoconstrictor and cardiac chronotropic responses to exogenous bolus doses of phenylephrine, angiotensin II and arginine vasopressin were recorded. Fetal cardiac baroreflex slopes were obtained using the peak pressor and heart rate responses to increasing doses of phenylephrine. Fetal treatment with phenylephrine, angiotensin II and vasopressin produced significant changes in arterial blood pressure, hind limb vascular resistance and heart rate. Pressor and vasopressor responses to all agonists were greater at 0.9 than at 0.6 of gestation; however, fetal cardiac baroreflex sensitivity decreased with advancing gestational age. Correlation analysis revealed that fetal plasma cortisol rather than gestational age was a greater determinant of pressor and vasopressor reactivity. In contrast, gestational age rather than cortisol better determined heart rate and baroreflex responsiveness in the equine fetus. The data show that development of cardiovascular function in the equine fetus occurs via cortisol dependent and -independent pathways. PMID- 16469781 TI - Placental adaptive responses and fetal programming. AB - Fetal programming occurs when the normal pattern of fetal development is disrupted by an abnormal stimulus or 'insult' applied at a critical point in in utero development. This then leads to an effect, for example diabetes or hypertension, which manifests itself in adult life. As the placenta is the regulator of nutrient composition and supply from mother to fetus and the source of hormonal signals that affect maternal and fetal metabolism, appropriate development of the placenta is crucial to normal fetal development. Placental function evolves in a carefully orchestrated developmental cascade throughout gestation. Disruption of this cascade can lead to abnormal development of the placental vasculature or of the trophoblast. Timing of a developmental 'insult' will be critical in consequent placental function and hence programming of the fetus. The 'insults' that alter placental development include hypoxia and abnormal maternal nutrient status, to which the placenta may adapt by alterations in transporter expression and activity to maintain fetal growth or by epigenetic regulation of placental gene expression. Hypoxia is physiological for organogenesis and placental tissue normally exists in a relatively hypoxic environment, but intrauterine growth restriction (IUGR) and pre-eclampsia are associated with a greater degree of trophoblast hypoxia. The metabolic activity of placental mitochondria leads to oxidative stress even in normal pregnancy which is exacerbated further in IUGR, diabetic and pre-eclamptic pregnancies and may also give nitrative stress known to lead to covalent modification and hence altered activity of proteins. Hypoxia, oxidative and nitrative stress all alter placenta development and may be a general underlying mechanism that links altered placental function to fetal programming. PMID- 16469780 TI - Fetal programming of hypothalamo-pituitary-adrenal function: prenatal stress and glucocorticoids. AB - Prenatal stress (PS) and maternal exposure to exogenous glucocorticoids can lead to permanent modification of hypothalamo-pituitary-adrenal (HPA) function and stress-related behaviour. Both of these manipulations lead to increased fetal exposure to glucocorticoids. Glucocorticoids are essential for many aspects of normal brain development, but exposure of the fetal brain to an excess of glucocorticoids can have life-long effects on neuroendocrine function. Both endogenous glucocorticoid and synthetic glucocorticoid exposure have a number of rapid effects in the fetal brain, including modification of neurotransmitter systems and transcriptional machinery. Such fetal exposure permanently alters HPA function in prepubertal, postpubertal and ageing offspring, in a sex-dependent manner. Prenatal stress and exogenous glucocorticoid manipulation also lead to the modification of behaviour, brain and organ morphology, as well as altered regulation of other endocrine systems. It is also becoming increasingly apparent that the timing of exposure to PS or synthetic glucocorticoids has tremendous effects on the nature of the phenotypic outcome. Permanent changes in endocrine function will ultimately impact on health in both human and animal populations. PMID- 16469782 TI - Activity-dependent regulation of synaptic strength and neuronal excitability in central auditory pathways. AB - Neural activity plays an important role in regulating synaptic strength and neuronal membrane properties. Attempts to establish guiding rules for activity dependent neuronal changes have led to such concepts as homeostasis of cellular activity and Hebbian reinforcement of synaptic strength. However, it is clear that there are diverse effects resulting from activity changes, and that these changes depend on the experimental preparation, and the developmental stage of the neural circuits under study. In addition, most experimental evidence on activity-dependent regulation comes from reduced preparations such as neuronal cultures. This review highlights recent results from studies of the intact mammalian auditory system, where changes in activity have been shown to produce alterations in synaptic and membrane properties at the level of individual neurons, and changes in network properties, including the formation of tonotopic maps. PMID- 16469783 TI - Evidence for altered placental blood flow and vascularity in compromised pregnancies. AB - The placenta is the organ that transports nutrients, respiratory gases, and wastes between the maternal and fetal systems. Consequently, placental blood flow and vascular development are essential components of normal placental function and are critical to fetal growth and development. Normal fetal growth and development are important to ensure optimum health of offspring throughout their subsequent life course. In numerous sheep models of compromised pregnancy, in which fetal or placental growth, or both, are impaired, utero-placental blood flows are reduced. In the models that have been evaluated, placental vascular development also is altered. Recent studies found that treatments designed to increase placental blood flow can 'rescue' fetal growth that was reduced due to low maternal dietary intake. Placental blood flow and vascular development are thus potential therapeutic targets in compromised pregnancies. PMID- 16469784 TI - Effects of secreted oligomers of amyloid beta-protein on hippocampal synaptic plasticity: a potent role for trimers. AB - The accumulation of amyloid beta-protein (Abeta) in brain regions serving memory and cognition is a central pathogenic feature of Alzheimer's disease (AD). We have shown that small soluble oligomers of human Abeta that are naturally secreted by cultured cells inhibit hippocampal long-term potentiation (LTP) in vitro and in vivo and transiently impair the recall of a complex learned behaviour in rats. These results support the hypothesis that diffusible oligomers of Abeta initiate a synaptic dysfunction that may be an early event in AD. We now report detailed electrophysiological analyses that define conditions under which acute application of soluble Abeta inhibits hippocampal synaptic plasticity in wild-type mice. To ascertain which Abeta assemblies contribute to the impairment of LTP, we fractionated oligomers by size-exclusion chromatography and found that Abeta trimers fully inhibit LTP, whereas dimers and tetramers have an intermediate potency. Natural Abeta oligomers are sensitive to heat denaturation, primarily inhibit the induction phase of LTP, and cause a sustained impairment of LTP even after extensive washout. We observed no effects of Abeta oligomers on presynaptic vesicle release. LTP in juvenile mice is resistant to the effects of Abeta oligomers, as is brain-derived-neurotrophic-factor-induced LTP in adult hippocampus. We conclude that specific assemblies, particularly timers, of naturally secreted Abeta oligomers are potent and selective inhibitors of certain forms of hippocampal LTP. PMID- 16469785 TI - Calcium-sensing mechanism in TRPC5 channels contributing to retardation of neurite outgrowth. AB - The calcium- and sodium-permeable transient receptor potential channel TRPC5 has an inhibitory role in neuronal outgrowth but the mechanisms governing its activity are poorly understood. Here we propose a mechanism involving the neuronal calcium sensor-1 (NCS-1) protein. Inhibitory mutants of TRPC5 and NCS-1 enhance neurite outgrowth similarly. Mutant NCS-1 does not inhibit surface expression of TRPC5 but generally suppresses channel activity, irrespective of whether it is evoked by carbachol, store depletion, lanthanides or elevated intracellular calcium. NCS-1 and TRPC5 are in the same protein complex in rat brain and NCS-1 directly binds to the TRPC5 C-terminus. The data suggest protein protein interaction between NCS-1 and TRPC5, and involvement of this protein complex in retardation of neurite outgrowth. PMID- 16469787 TI - Muscle metaboreflex contribution to cardiovascular regulation during dynamic exercise in microgravity: insights from mission STS-107 of the space shuttle Columbia. AB - One of the most important features of prolonged weightlessness is a progressive impairment of muscular function with a consequent decrease in exercise capacity. We tested the hypothesis that the impairment in musculo-skeletal function that occurs in microgravity results in a potentiation of the muscle metaboreflex mechanism and also affects baroreflex modulation of heart rate (HR) during exercise. Four astronauts participating in the 16 day Columbia shuttle mission (STS-107) were studied 72-71 days before launch and on days 12-13 in-flight. The protocol consisted of 6 min bicycle exercise at 50% of individual V(o2,max) followed by 4 min of postexercise leg circulatory occlusion (PECO). At rest, systolic (S) and diastolic (D) blood pressure (BP), R-R interval and baroreflex sensitivity (BRS) did not differ significantly between pre- and in-flight measurements. Both pre- and in-flight, SBP increased and R-R interval and BRS decreased during exercise, whereas DBP did not change. During PECO preflight, SBP and DBP were higher than at rest, whereas R-R interval and BRS recovered to resting levels. During PECO in-flight, SBP and DBP were significantly higher whereas R-R interval and BRS remained significantly lower than at rest. The part of the SBP response (delta) that was maintained by PECO was significantly greater during spaceflight than before (34.5 +/- 8.8 versus 13.8 +/- 11.9 mmHg, P = 0.03). The tachycardic response to PECO was also significantly greater during spaceflight than preflight (-141.5 +/- 25.2 versus - 90.5 +/- 33.3 ms, P = 0.02). This study suggests that the muscle metaboreflex is enhanced during dynamic exercise in space and that the potentiation of the muscle metaboreflex affects the vagally mediated arterial baroreflex contribution to HR control. PMID- 16469786 TI - Mechanisms of CO2/H+ chemoreception by respiratory rhythm generator neurons in the medulla from newborn rats in vitro. AB - We investigated mechanisms of CO(2)/H(+) chemoreception in the respiratory centre of the medulla by measuring membrane potentials of pre-inspiratory neurons, which are putative respiratory rhythm generators, in the brainstem-spinal cord preparation of the neonatal rat. Neuronal response was tested by changing superfusate CO(2) concentration from 2% to 8% at constant HCO(3)(-) concentration (26 mm) or by changing pH from 7.8 to 7.2 by reducing HCO(3)(-) concentration at constant CO(2) (5%). Both respiratory and metabolic acidosis lead to depolarization of neurons with increased excitatory synaptic input and increased burst rate. Respiratory acidosis potentiated the amplitude of the neuronal drive potential. In the presence of tetrodotoxin (TTX), membrane depolarization persisted during respiratory and metabolic acidosis. However, the depolarization was smaller than that before application of TTX, which suggests that some neurons are intrinsically, and others synaptically, chemosensitive to CO(2)/H(+). Application of Ba(2+) blocked membrane depolarization by respiratory acidosis, whereas significant depolarization in response to metabolic acidosis still remained after application of Cd(2+) and Ba(2+). We concluded that the intrinsic responses to CO(2)/H(+)changes were mediated by potassium channels during respiratory acidosis, and that some other mechanisms operate during metabolic acidosis. In low-Ca(2+), high-Mg(2+) solution, an increased CO(2) concentration induced a membrane depolarization with a simultaneous increase of the burst rate. Pre-inspiratory neurons could adapt their baseline membrane potential to external CO(2)/H(+) changes by integration of these mechanisms to modulate their burst rates. Thus, pre-inspiratory neurons might play an important role in modulation of respiratory rhythm by central chemoreception in the brainstem-spinal cord preparation. PMID- 16469788 TI - Removal of gating in voltage-dependent ClC-2 chloride channel by point mutations affecting the pore and C-terminus CBS-2 domain. AB - Functional and structural studies demonstrate that Cl(-) channels of the ClC family have a dimeric double-barrelled structure, with each monomer contributing an identical pore. Studies with ClC-0, the prototype ClC channel, show the presence of independent mechanisms gating the individual pores or both pores simultaneously. A single-point mutation in the CBS-2 domain of ClC-0 has been shown to abolish slow gating. We have taken advantage of the high conservation of CBS domains in ClC channels to test for the presence of a slow gate in ClC-2 by reproducing this mutation (H811A). ClC-2-H811A showed faster opening kinetics and opened at more positive potentials than ClC-2. There was no difference in [Cl( )](i) dependence. Additional neutralization of a putative pore gate glutamate side chain (E207V) abolished all gating. Resolving slow and fast gating relaxations, however, revealed that the H811A mutation affected both fast and slow gating processes in ClC-2. This suggests that slow and fast gating in ClC-2 are coupled, perhaps with slow gating contributing to the operation of the pore E207 as a protopore gate. PMID- 16469789 TI - Propriospinal neurons contribute to bulbospinal transmission of the locomotor command signal in the neonatal rat spinal cord. AB - This study examines whether propriospinal transmission contributes to descending propagation of the brainstem locomotor command signal in the in vitro neonatal rat spinal cord. Using double bath partitions, synaptic transmission was suppressed in the cervicothoracic region while monitoring locomotor-like activity on lumbar ventral roots evoked by either chemical or electrical stimulation of the brainstem. Locomotor-like activity induced by electrical stimulation was more stable (cycle period coefficient of variation (CV) 11.7 +/- 6.1%) than the rhythm induced by chemical stimulation (CV 31.3 +/- 6.4%). Ca(2+)-free bath solution, elevated Mg(2+) ion concentration, excitatory amino acid receptor antagonists (AP5 and/or CNQX), and the muscarinic receptor antagonist, atropine, were used in attempts to block synaptic transmission. Each of these manipulations, except muscarinic receptor blockade, was capable of blocking locomotor-like activity induced by brainstem stimulation. However, locomotor-like activity induced by higher intensity electrical stimulation of the brainstem (1.2-5 times threshold) was relatively refractory to synaptic suppression using AP5 and CNQX, and Ca(2+) free solution was more effective if combined with high Mg(2+) (15 mm) or EGTA. Enhancement of neuronal excitation in the cervicothoracic region, using Mg(2+) free bath solution, facilitated brainstem activation of locomotor-like activity in the lumbar cord, consistent with a propriospinal mechanism of locomotor signal propagation. Blockade of brainstem-induced locomotor-like activity was related to the number of cervicothoracic segments exposed to synaptic suppression, being most effective if five or more segments were included. These results provide direct evidence that propriospinal pathways contribute to bulbospinal activation of the locomotor network in the in vitro neonatal rat brainstem-spinal cord preparation, and suggest that a propriospinal system is recruited in parallel with long direct projections that activate the locomotor network. PMID- 16469791 TI - Does a negative screening colonoscopy ever need to be repeated? AB - BACKGROUND AND AIMS: Screening colonoscopy is thought to be a powerful and cost effective tool to reduce colorectal cancer incidence and mortality. Whether and when colonoscopy with negative findings has to be repeated is not well defined. The aim of this study was to assess the long term risk of clinically manifest colorectal cancer among subjects with negative findings at colonoscopy. PATIENTS: 380 cases and 485 controls participating in a population based case-control study in Germany. METHODS: Detailed history and results of previous colonoscopies were obtained by interview and from medical records. Adjusted relative risks of colorectal cancer among subjects with a previous negative colonoscopy compared with those without previous colonoscopy were estimated according to time since colonoscopy. RESULTS: Subjects with previous negative colonoscopy had a 74% lower risk of colorectal cancer than those without previous colonoscopy (adjusted odds ratio (aOR) = 0.26 (95% confidence interval, 0.16 to 0.40)). This low risk was seen even if the colonoscopy had been done up to 20 or more years previously. Particularly low risks were seen for sigma cancer (aOR = 0.13 (0.04 to 0.43)) and for rectal cancer (aOR = 0.19 (0.09 to 0.39)), and after a negative screening colonoscopy at ages 55 to 64 (aOR = 0.17 (0.08 to 0.39)) and older (aOR = 0.21 (0.10 to 0.41)). CONCLUSIONS: Subjects with negative findings at colonoscopy are at very low risk of colorectal cancer and might not need to undergo repeat colonoscopy for 20 years or more, if at all. The possibility of extending screening intervals to 20 years or more might reduce complications and increase the feasibility and cost-effectiveness of colonoscopy based screening programmes. PMID- 16469790 TI - Tuning and playing a motor rhythm: how metabotropic glutamate receptors orchestrate generation of motor patterns in the mammalian central nervous system. AB - Repeated motor activities like locomotion, mastication and respiration need rhythmic discharges of functionally connected neurons termed central pattern generators (CPGs) that cyclically activate motoneurons even in the absence of descending commands from higher centres. For motor pattern generation, CPGs require integration of multiple processes including activation of ion channels and transmitter receptors at strategic locations within motor networks. One emerging mechanism is activation of glutamate metabotropic receptors (mGluRs) belonging to group I, while group II and III mGluRs appear to play an inhibitory function on sensory inputs. Group I mGluRs generate neuronal membrane depolarization with input resistance increase and rapid fluctuations in intracellular Ca(2+), leading to enhanced excitability and rhythmicity. While synchronicity is probably due to modulation of inhibitory synaptic transmission, these oscillations occurring in coincidence with strong afferent stimuli or application of excitatory agents can trigger locomotor-like patterns. Hence, mGluR-sensitive spinal oscillators play a role in accessory networks for locomotor CPG activation. In brainstem networks supplying tongue muscle motoneurons, group I receptors facilitate excitatory synaptic inputs and evoke synchronous oscillations which stabilize motoneuron firing at regular, low frequency necessary for rhythmic tongue contractions. In this case, synchronicity depends on the strong electrical coupling amongst motoneurons rather than inhibitory transmission, while cyclic activation of K(ATP) conductances sets its periodicity. Activation of mGluRs is therefore a powerful strategy to trigger and recruit patterned discharges of motoneurons. PMID- 16469792 TI - Abnormal oral mucosal light reflectance: a new clinical marker of high risk for colorectal cancer. AB - BACKGROUND: A familial predisposition to colorectal cancer (CRC) has been clearly established, consisting of familial clustering in 15-20% and clear hereditary aetiology in 5-10% of overall CRC cases. Early identification of families and individuals at high risk is essential as intensive surveillance has been demonstrated to reduce cancer incidence and overall mortality. In the present study, the value of oral mucosal light reflectance in identifying hereditary non polyposis colorectal cancer (HNPCC) carriers was investigated. METHODS: Twenty members of six different genetically unrelated HNPCC kindred and 30 genetically unrelated age and sex matched healthy controls were examined. Lower gingival and vestibular oral mucosal reflectance was measured using an imaging spectrophotometer. RESULTS: HNPCC carriers showed significantly lower values in the 590-700 nm wavelength range (p99.5%. CONCLUSIONS: These preliminary results show that elastic scattering spectroscopy has the potential to target conventional biopsies in Barrett's surveillance saving significant endoscopist and pathologist time with consequent financial savings. This technique now requires validation in prospective studies. PMID- 16469794 TI - Analysis of the influence of OCTN1/2 variants within the IBD5 locus on disease susceptibility and growth indices in early onset inflammatory bowel disease. AB - BACKGROUND AND AIMS: The OCTN1 (SLC22A4 1672C-->T) and OCTN2 (SLC22A5 -207G-->C) variants within the IBD5 locus have been associated with susceptibility to adult onset Crohn's disease (CD), but their contribution in children has not been examined. METHODS: These OCTN1/2 variants and IBD5 marker single nucleotide polymorphisms (SNPs) (IGR2096a_1, IGR2198a_1, and IGR2230a_1) were examined in 299 Scottish children (200 with CD, 74 with ulcerative colitis (UC), and 25 with indeterminate colitis (IC)), together with 502 parents (for transmission disequilibrium testing) and 256 controls. RESULTS: All SNPs were in strong linkage disequilibrium (D' >0.94). TDT analysis showed association of the OCTN1 variant with inflammatory bowel disease (IBD) (p = 0.01) and CD (p = 0.04). Allele frequencies of the OCTN1/2 variants were significantly higher in IBD/CD cases (p<0.04). The homozygous mutant OCTN1/2 haplotype was increased in IBD (24.3% v 16.1%, p = 0.02) and UC (28.2% v 16.1%, p = 0.02) compared with controls. The OCTN1/2 variants were not independent of the background IBD5 risk haplotype in conferring disease susceptibility. Unifactorial analysis in CD patients showed that carriage of the TC haplotype was associated with lower weight, height, and BMI centile (<9(th) centile) at diagnosis (weight: 87.9% v 67.3% (p = 0.002), odds ratio (OR) = 3.52 (95% confidence interval, 1.51 to 8.22); height: 84.1% v 68.4% (p<0.05), OR = 2.44 (1.00 to 5.99); BMI: 79.6% v 61.1% (p = 0.02), OR = 2.49 (1.14 to 5.44)), and lower weight centile at follow up (87.5% v 64.6% (p = 0.03), OR = 3.83 (1.03 to 14.24)). Multifactorial binary logistic regression analysis confirmed association of the TC haplotype with lower weight centile at diagnosis (p = 0.02, OR = 3.41 (1.20 to 9.66)). CONCLUSIONS: These data implicate variants within the IBD5 haplotype, as determinants of disease susceptibility and growth indices in early onset IBD. The OCTN1/2 variants remain potential positional candidate genes, but require further analysis. PMID- 16469797 TI - Experience-induced changes in taste identification of monosodium glutamate (MSG) are reversible. AB - A few studies have reported experience-inducible changes in human taste and olfactory sensitivities. However, no study thus far has systematically characterized the stability of the enhanced sensitivities. In our previous study, we found increases in taste identification ability for monosodium glutamate (MSG) in subjects who had been briefly exposed to MSG in food for 10 days. Here, we tested the temporal stability of the enhanced taste identification ability. First, we exposed a group of 20 subjects to MSG in food and then compared their sensitivities to MSG with those of a control group. When tested on day 11 or 12, the mean MSG taste identification ability of the MSG-exposed group was significantly higher than the control group. Next, 11 of the subjects who were exposed to MSG in food initially, and then stopped being exposed performed significantly poorer in identifying MSG after 10 days of the nonexposure than they did 10 days before. In contrast, nine subjects who were exposed to MSG initially and continued being exposed maintained their high identification levels. These results support earlier finding of the plasticity in the taste identification of MSG and show that the enhanced identification ability can be reversed rapidly when MSG exposure is not sustained. PMID- 16469798 TI - Enhancement of cortisol-induced 11beta-hydroxysteroid dehydrogenase type 1 expression by interleukin 1beta in cultured human chorionic trophoblast cells. AB - Chorion is the most abundant site of 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) expression within intrauterine tissues. It is important to study the regulation of 11beta-HSD1 expression in the chorion in terms of local cortisol production during pregnancy. Using real-time PCR and enzyme activity assay, we found that cortisol (1 mum) and IL-1beta (10 ng/ml) for 24 h significantly increased 11beta-HSD1 mRNA expression and reductase activity in cultured human chorionic trophoblasts. A further significant increase of 11beta HSD1 mRNA expression and reductase activity was observed with cotreatment of cortisol and IL-1beta. To explore the mechanism of induction, 11beta-HSD1 promoter was cloned into pGL3 plasmid expressing a luciferase reporter gene. By transfecting the constructed vector into WISH cells, an amnion-derived cell line, we found that cortisol (1 microM) or IL-1beta (10 ng/ml) significantly increased reporter gene expression. Likewise, an additional increase in reporter gene expression was observed with cotreatment of cortisol and IL-beta. To explore the physiological significance of 11beta-HSD1 induction in the chorion, we studied the effect of cortisol on cytosolic phospholipase A(2) and cyclooxygenase 2 expression. We found that treatment of chorionic trophoblast cells with cortisol (1 microM) induced both cytosolic phospholipase A(2) and cyclooxygenase 2 mRNA expression. We conclude that cortisol up-regulates 11beta-HSD1 expression through induction of promoter activity, and the effect was enhanced by IL-1beta, suggesting that more biologically active glucocorticoids could be generated in the fetal membranes in the presence of infection, which may consequently feed forward in up-regulation of prostaglandin synthesis. PMID- 16469799 TI - Continuous human metastin 45-54 infusion desensitizes G protein-coupled receptor 54-induced gonadotropin-releasing hormone release monitored indirectly in the juvenile male Rhesus monkey (Macaca mulatta): a finding with therapeutic implications. AB - The effect of continuous administration of the C-terminal fragment of metastin, the ligand for the G protein-coupled receptor, GPR54, on GnRH-induced LH secretion was examined in three agonadal, juvenile male monkeys whose responsiveness to GnRH was heightened by pretreatment with a chronic pulsatile iv infusion of synthetic GnRH. After bolus injection of 10 microg human (hu) metastin 45-54 (equivalent to kisspeptin 112-121), the GPR54 agonist was infused continuously at a dose of 100 microg/h and elicited a brisk LH response for approximately 3 h. This rise was then followed by a precipitous drop in LH despite continuous exposure of GPR54 to metastin 45-54. On d 4, during the final 3 h of the infusion, single boluses of hu metastin 45-54 (10 microg), N-methyl-DL aspartic acid (NMDA) (10 mg/kg) and GnRH (0.3 microg) were administered to interrogate each element of the metastin-GPR54-GnRH-GnRH receptor cascade. Although the NMDA and GnRH boluses were able to elicit LH pulses, that of hu metastin 45-54 was not, demonstrating functional integrity of GnRH neurons (NMDA) and GnRH receptors (NMDA and GnRH) but desensitization of GPR54. The desensitization of GPR54 by continuous hu metastin 45-54 administration has therapeutic implications for a variety of conditions currently being treated by GnRH and its analogs, including restoration of fertility in patients with abnormal GnRH secretion (i.e. idiopathic hypogonadotropic hypogonadism and hypothalamic amenorrhea) and selective, reversible suppression of the pituitary gonadal axis to achieve suppression of gonadal steroids (i.e. precocious puberty, endometriosis, uterine fibroids, and prostate cancer). PMID- 16469800 TI - Identification of the chicken growth hormone-releasing hormone receptor (GHRH-R) mRNA and gene: regulation of anterior pituitary GHRH-R mRNA levels by homologous and heterologous hormones. AB - GHRH stimulates GH secretion in chickens as in mammals. However, nothing is known about the chicken GHRH receptor (GHRH-R). Here we report the cDNA sequence of chicken GHRH-R. Comparison of the cDNA sequence with the chicken genome localized the GHRH-R gene to chicken chromosome 2 and indicated that the chicken GHRH-R gene consists of 13 exons. Expression of all exons was confirmed by RT-PCR amplification of pituitary mRNA. The amino acid sequence predicted by the GHRH-R cDNA is homologous to that in other vertebrates and contains seven transmembrane domains and a conserved hormone-binding domain. The predicted size of the GHRH-R protein (48.9 kDa) was confirmed by binding of (125)I-GHRH to chicken pituitary membranes and SDS-PAGE. GHRH-R mRNA was readily detected by RT-PCR in the pituitary but not in the hypothalamus, total brain, lung, adrenal, ovary, or pineal gland. Effects of corticosterone (CORT), GHRH, ghrelin, pituitary adenylate cyclase-activating peptide, somatostatin (SRIF), and TRH on GHRH-R and GH gene expression were determined in cultures of chicken anterior pituitary cells. GHRH-R and GH mRNA levels were determined by quantitative real-time RT PCR. Whereas all treatments affected levels of GH mRNA, only CORT, GHRH, and SRIF significantly altered GHRH-R mRNA levels. GHRH-R gene expression was modestly increased by GHRH and suppressed by SRIF at 4 h, and CORT dramatically decreased levels of GHRH-R mRNA at 72 h. We conclude that adrenal glucocorticoids may substantially impact pituitary GH responses to GHRH in the chicken through modulation of GHRH-R gene expression. PMID- 16469801 TI - Activation of p70S6K induces expression of matrix metalloproteinase 9 associated with hepatocyte growth factor-mediated invasion in human ovarian cancer cells. AB - The expression of hepatocyte growth factor (HGF) receptor, encoded by the Met oncogene, is elevated in ovarian and a variety of cancers. Here we show that human ovarian cancer cells with high Met expression were more sensitive to the cell motility and invasion effect of HGF. Met down-regulation by small interfering RNAs or K252a resulted in reduced migration in response to HGF. The invasive/migratory phenotype activated by HGF can be blocked by specific inhibitors of the phosphatidylinositol-3-kinase (PI3K) cascade, inhibitor of p70(S6K), and also the expression of a dominant-negative Akt, demonstrating that HGF transmits the motogenic signal through PI3K and Akt to p70(S6K). A significant role for p70(S6K) in cell invasion is further supported by the observation that expression of constitutively active forms of p70(S6K) is sufficient to induce invasive and migratory phenotypes in ovarian cancer cells. Importantly, activation of p70(S6K) stimulated expression and proteolytic activity of matrix metalloproteinase (MMP)-9 and cellular invasion, whereas it had little effect on MMP-2, suggesting for the first time that MMP-9 up regulation by p70(S6K) as a key step for HGF-induced invasion and migration. These data suggest that interfering p70(S6K) may provide a novel means of controlling tumor cell invasiveness. PMID- 16469802 TI - Effect of interferon-tau administration on endometrium of nonpregnant ewes: a comparison with pregnant ewes. AB - In ruminants, conceptus interferon-tau (IFNT) alters maternal physiology to accommodate a pregnancy. We hypothesized that the effectiveness of IFNT on extending corpus luteum (CL) life span in nonpregnant ewes would depend upon the dose and manner of administration and would be correlated with the response in gene expression in endometrium. We anticipated that IFNT, whether administered im or by uterine infusion, would mimic changes observed in pregnancy. Ewes were assigned to five treatments: 1) uterine infusion of saline; 2) uterine infusion of ovine IFNT4 (200 microg/d); 3) saline im injection; 4) im injection of IFNT4 at low dose (200 microg/d); and 5) high dose (2 mg/d). CL life span was increased in groups 2 and 5, but not in 1, 3, and 4. Endometrial RNA extracted from groups 1-5 on d 14 and from d 14 pregnant and nonbred (cyclic) ewes was used to assess expression of 70 genes on microarrays. When pregnant and cyclic ewes were compared, 30 genes were up-regulated and nine down-regulated during pregnancy. Responses were slightly less in groups 2 and 5 but were much lower in group 4. The majority of the highly up-regulated genes were associated with antiviral responses. Those down-regulated included ones for IGF-II, hypoxia-inducible factor 1alpha, oxytocin receptor, prostaglandin F synthase, and cyclooxygenase-2. Quantitative PCR for selected genes confirmed these data and revealed that similar gene expression changes occurred in the CL of pregnant and group 2 ewes. IFNT treatment mimics pregnancy, but relatively high doses of im-injected IFNT are required to elicit a full endometrial response. PMID- 16469804 TI - Dynamic nongenomic actions of thyroid hormone in the developing rat brain. AB - Two well-characterized nongenomic actions of thyroid hormone in cultured brain tissues are: 1) regulation of type 2 iodothyronine 5'deiodinase (D2) activity and 2) regulation of actin polymerization. In particular, the latter is likely to have profound effects on neuronal migration in the developing brain. In this study, we determined whether these nongenomic actions also occurred in vivo during brain development. Neonatal hypothyroidism was induced by propylthiouracil given to pregnant dams beginning on d17 of gestation and continued throughout the neonatal period. On postnatal d 14, rats were injected with either cold or [(125)I]-labeled iodothyronines and killed sequentially after injection. In contrast to reports in the adult rat, all three iodothyronines readily and equally entered developing brain tissues. As expected, cerebrocortical D2 activity was markedly elevated in the hypothyroid brain and both reverse T(3) (rT(3)) and T(4) rapidly decreased D2 to euthyroid levels within 3 h. Furthermore, cerebellar G-actin content in the hypothyroid rat was approximately 5-fold higher than in the euthyroid rat. Again, both rT(3) and T(4) rapidly decreased the G-actin content by approximately 50%, with a reciprocal increase in F-actin content to euthyroid levels without altering total actin. Neither T(3) nor vehicle had any effect on D2 activity in the cortex or G- or F-actin content in the cerebellum. The thyroid hormone-dependent regulation of actin polymerization in the rat brain provides a mechanism by which this morphogenic hormone can influence neuronal migration independent of the need for altered gene transcription. Furthermore, these data suggest a prominent role for rT(3) during brain development. PMID- 16469803 TI - Estren behaves as a weak estrogen rather than a nongenomic selective activator in the mouse uterus. AB - A proposed membrane-mediated mechanism of rapid nongenomic response to estrogen has been the intense focus of recent research. Estren, a synthetic steroid, is reported to act selectively through a rapid membrane-mediated pathway, rather than through the classical nuclear estrogen receptor (ER)-mediated pathway, to maintain bone density in ovariectomized mice without uterotropic effects. To evaluate the mechanism and physiological effects of estren, we studied responses in adult ovariectomized mice. In a 3-d uterine bioassay, we found that 300 microg estren significantly increased uterine weight; in comparison, a more maximal response was seen with 1 mug estradiol (E2). The estren response was partly ERalpha independent, because ERalpha knockout (alphaERKO) uteri also exhibited a more moderate weight increase. Estren induced epithelial cell proliferation in wild-type, but not alphaERKO, mice, indicating ERalpha dependence of the epithelial growth response. Examination of estren-regulated uterine genes by microarray indicated that early (2 h) changes in gene expression are similar to the early responses to E2. These gene responses are ERalpha dependent, because they are not seen in alphaERKO mice. Later estren-induced changes in gene expression (24 h) are blunted compared with those seen 24 h after E2. In contrast to early genes, these later estren responses are independent of ERalpha, because the alphaERKO shows a similar response to estren at 24 h. We found that E2 or estren treatments lead to depletion of ERalpha in the uterine cytosol fraction and accumulation in the nuclear fraction within 30-60 min, consistent with the ability of estren to regulate genes through a nuclear ERalpha rather than a nongenomic mechanism. Interestingly, estren, but not E2, induces accumulation of androgen receptor (AR) in the nuclear fraction of both wild-type and alphaERKO samples, suggesting that AR might be involved in the later ERalpha-independent genomic responses to estren. In conclusion, our studies suggest that estren is weakly estrogenic in the mouse uterus and might induce nuclear ERalpha- and AR mediated responses. Given its activity in our uterine model, the use of estren as a bone-selective clinical compound needs to be reconsidered. PMID- 16469805 TI - Insulin-like growth factor (IGF) binding protein-3 attenuates prostate tumor growth by IGF-dependent and IGF-independent mechanisms. AB - IGF binding protein (IGFBP)-3 inhibits cell growth and promotes apoptosis by sequestering free IGFs. In addition IGFBP-3 has IGF-independent, proapoptotic, antiproliferative effects on prostate cancer cells in vitro. Expression of the large T-antigen (Tag) under the long probasin promoter (LPB) in LPB-Tag mice results in prostate tumorigenesis. To investigate the IGF-dependent and IGF independent effects of IGFBP-3 on prostate tumor growth, we crossed LPB-Tag mice with cytomegalovirus (CMVBP-3) and phosphoglycerate kinase (PGKBP-3) mice that overexpress IGFBP-3 under the cytomegalovirus promoter and the phosphoglycerate kinase promoter, respectively, and also I56G/L80G/L81G-mutant IGFBP-3 (PGKmBP-3) mice that express I56G/L80G/L81G-IGFBP-3, a mutant, that does not bind IGF-I but retains IGF-independent proapoptotic effects in vitro. Prostate tumor size and the steady-state level of p53 were attenuated in LPB-Tag/CMVBP-3 and LPB Tag/PGKBP-3 mice, compared with LPB-Tag/wild-type (Wt) mice. A more marked effect was observed in LPB-Tag/CMVBP-3, compared with LPB-Tag/PGKBP-3, reflecting increased levels of transgene expression in CMVBP-3 prostate tissue. No attenuation of tumor growth was observed in LPB-Tag/PGKmBP-3 mice during the early tumor development, indicating that the inhibitory effects of IGFBP-3 were most likely IGF dependent during the initiation of tumorigenesis. At 15 wk of age, epidermal growth factor receptor expression was increased in LPB-Tag/Wt and LPB-Tag/PGKmBP-3 tissue, compared with LPB-Tag/PGKBP-3. IGF receptor was increased in all transgenic mice, but pAkt expression, a marker of downstream IGF I action, was increased only in LPB-Tag/Wt and LPB-Tag/PGKmBP-3. After 15 wk of age, a marked reduction in tumor growth was apparent in LPB-Tag/PGKmBP-3 mice, indicating that the IGF-independent effects of IGFBP-3 may be important in inhibiting tumor progression. PMID- 16469806 TI - Vasoactive intestinal polypeptide regulates dynamic changes in astrocyte morphometry: impact on gonadotropin-releasing hormone neurons. AB - Recent studies suggest that astrocytes modulate the GnRH-induced LH surge. In particular, we have shown that the surface area of astrocytes that ensheath GnRH neurons exhibits diurnal rhythms. Vasoactive intestinal polypeptide (VIP) influences numerous aspects of astrocyte function in multiple brain regions and is a neurotransmitter in the suprachiasmatic nucleus (SCN) that affects GnRH neurons. The goals of this study were to: 1) assess whether astrocytes that surround GnRH neurons express VIP receptors, 2) determine the effects VIP suppression in the SCN on the morphometry of astrocytes surrounding GnRH neurons, and 3) assess whether this effect mimics aging-like changes in surface area of astrocytes. Young rats were ovariectomized (d 0), implanted with cannulae into the SCN (d 5), injected with VIP antisense (antioligo) or random sequence oligonucleotides, implanted with capsules containing 17beta-estradiol dissolved in oil (d 7), and perfused at 0300, 1400, and 1800 h (d 9). Brains were processed for immunocytochemistry. Our results demonstrate that astrocytes in close apposition to GnRH neurons express VIP receptors. Antioligo treatment blocked diurnal rhythms in surface area of astrocytes ensheathing GnRH neurons. The absence of diurnal rhythms resembles observations in middle-aged rats. Together these findings suggest that the ability of the VIP-containing neurons in the SCN to relay diurnal information to GnRH neurons may be by influencing dynamic changes in the morphometry of astrocytes that surround GnRH neurons. Furthermore, the absence of a VIP rhythm in aging animals may lead to altered GnRH activity via astrocyte-dependent mechanisms. PMID- 16469807 TI - Leptin is required for uncoupling protein-1-independent thermogenesis during cold stress. AB - We investigated the role of leptin in regulating energy metabolism through induction of uncoupling protein (UCP)-1-based brown fat thermogenesis by comparing phenotypes of energy balance in ob/ob and double-mutant ob/ob.Ucp1(-/-) mice. Measurements of adiposity and lean body mass (nuclear magnetic resonance), energy expenditure (indirect calorimetry), body weight, food intake, and core body temperature were determined in the two mutant stocks of 3-month-old mice maintained at an initial ambient temperature of 28 C for 21 d and then at 21 C for 16 d, and finally with leptin administration for 8 d at 21 C. No phenotypic differences between ob/ob and ob/ob.Ucp1(-/-) mice were detected, suggesting that UCP1-based thermogenesis is not essential for the regulation of adiposity in ob/ob mice at temperatures between 21 and 28 C. Although both Ucp1(-/-) and ob/ob mice can survive in extreme cold at 4 C, provided they are adapted to the cold by gradually lowering ambient temperature, ob/ob.Ucp1(-/-) mice could not adapt and survive at temperatures lower than 12 C unless they were administered leptin. As the ambient temperature was reduced from 20 to 16 C, ob/ob.Ucp1(-/-) mice treated with leptin have elevated levels of circulating T(3) that correlate with elevated sarcoendoplasmic reticulum Ca(2+) ATPase 2a mRNA levels in gastrocnemius muscle. Furthermore, ob/ob.Ucp1(-/-) mice, treated with T(3), were able to maintain body temperature and stimulate sarcoendoplasmic reticulum Ca(2+) ATPase 2a expression when the ambient temperature was gradually reduced to 4 C. Thus, in the absence of UCP1, leptin-induced thermogenesis protects body temperature in part through its action on the thyroid hormone axis. PMID- 16469809 TI - Brain uptake of intranasally applied radioiodinated leptin in Wistar rats. AB - Leptin is mainly synthesized and secreted by fat cells in proportion to adipose tissue mass. Under physiological conditions, this hormone reduces food intake and increases thermogenesis through interactions with neurons in the central nervous system. However, transport of leptin into the central nervous system via the blood-brain barrier is saturable, and in obesity the feedback signal to the brain is markedly insufficient. In recent experiments we have shown, that intranasal (i.n.) delivery of leptin reduces food intake in rats. The aim of the present study was to explore the distribution of i.n. delivered leptin within brain, blood, and peripheral tissues. Application of [(125)I]leptin (0.03, 0.1, and 0.2 mg/kg) into male Wistar rats' nares (n = 8 per group) leads to supraphysiological brain leptin concentrations 30 min after application, with contents in the hypothalamus (7.3 +/- 2.6, 5.9 +/- 1.6, and 13.8 +/- 5.7 ng/g; P = 0.023; F = 6.157) being significantly higher than the brain average (1.2 +/- 0.2, 3.9 +/- 1.0, and 6.0 +/- 1.9 ng/g). In contrast, contents in the occipital/entorhinal cortex were lower than the brain average, indicating a minor participation of the transport via cerebrospinal fluid, which would have favored cerebrospinal fluid exposed surfaces. In experiments employing the application of unlabeled leptin administered iv, we were able to show that excess blood leptin does not diminish brain uptake of i.n. leptin (as indicated by [(125)I]leptin), supporting a direct transport from nose to brain by circumvention of the blood-brain barrier. This study thus clearly demonstrates a rapid and highly effective transport of leptin from nose to brain. PMID- 16469808 TI - Diet-genotype interactions in the development of the obese, insulin-resistant phenotype of C57BL/6J mice lacking melanocortin-3 or -4 receptors. AB - Loss of brain melanocortin receptors (Mc3rKO and Mc4rKO) causes increased adiposity and exacerbates diet-induced obesity (DIO). Little is known about how Mc3r or Mc4r genotype, diet, and obesity affect insulin sensitivity. Insulin resistance, assessed by insulin and glucose tolerance tests, Ser(307) phosphorylation of insulin receptor substrate 1, and activation of protein kinase B, was examined in control and DIO wild-type (WT), Mc3rKO and Mc4rKO C57BL/6J mice. Mc4rKO mice were hyperphagic and had increased metabolic efficiency (weight gain per kilojoule consumed) relative to WT; both parameters increased further on high-fat diet. Obesity of Mc3rKO was more dependent on fat intake, involving increased metabolic efficiency. Fat mass of DIO Mc3rKO and Mc4rKO was similar, although Mc4rKO gained weight more rapidly. Mc4rKO develop hepatic insulin resistance and severe hepatic steatosis with obesity, independent of diet. DIO caused further deterioration of insulin action in Mc4rKO of either sex and, in male Mc3rKO, compared with controls, associated with increased fasting insulin, severe glucose intolerance, and reduced insulin signaling in muscle and adipose tissue. DIO female Mc3rKO exhibited very modest perturbations in glucose metabolism and insulin sensitivity. Consistent with previous data suggesting impaired fat oxidation, both Mc3rKO and Mc4rKO had reduced muscle oxidative metabolism, a risk factor for weight gain and insulin resistance. Energy expenditure was, however, increased in Mc4rKO compared with Mc3rKO and controls, perhaps due to hyperphagia and metabolic costs associated with rapid growth. In summary, DIO affects insulin sensitivity more severely in Mc4rKO compared with Mc3rKO, perhaps due to a more positive energy balance. PMID- 16469811 TI - The endogenous estrogen status regulates microglia reactivity in animal models of neuroinflammation. AB - It has been previously demonstrated that 17beta-estradiol (E(2)) inhibits the response of microglia, the resident brain macrophages, to acute injuries in specific brain regions. We here show that the effect of E(2) in acute brain inflammation is widespread and that the hormone reduces the expression of inflammatory mediators, such as monocyte chemoattractant protein-1, macrophage inflammatory protein-2, and TNF-alpha, induced by lipopolysaccharide, demonstrating that microglia are a direct target of estrogen action in brain. Using the APP23 mice, an animal model of Alzheimer's disease reproducing chronic neuroinflammation, we demonstrate that ovary ablation increases microglia activation at beta-amyloid (Abeta) deposits and facilitates the progression of these cells toward a highly reactive state. Long-term administration of E(2) reverts the effects of ovariectomy and decreases microglia reactivity compared with control animals. In this animal model, these events do not correlate with a reduced number of Abeta deposits. Finally, we show that E(2) inhibits Abeta induced expression of scavenger receptor-A in macrophage cells, providing a mechanism for the effect of E(2) on Abeta signaling observed in the APP23 mice. Altogether, our observations reveal a substantial involvement of endogenous estrogen in neuroinflammatory processes and provide novel mechanisms for hormone action in the brain. PMID- 16469810 TI - Importance of uterine cell death, renewal, and their hormonal regulation in hamsters that show progesterone-dependent implantation. AB - This study was initiated to investigate the significance of uterine cell death and proliferation during the estrous cycle and early pregnancy and their correlation with sex steroids in hamsters where blastocyst implantation occurs in only progesterone-primed uteri. The results obtained in hamsters were also compared with mice where blastocyst implantation occurs in progesterone-primed uteri if estrogen is provided. Apoptotic cells in the uterus were detected by using terminal deoxynucleotide transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) technique. Uterine cell proliferation was determined by 5-bromo-2'-deoxyuridine labeling followed by immunohistochemistry and methyl tritiated [(3)H]thymidine labeling. Active caspase-3, an executor protein of cell death, expression was assayed by immunohistochemistry/immunofluorescence. Our results demonstrate that epithelial proliferation on the second day after mating marks the initiation of pregnancy-related uterine changes in both species despite their differences in hormonal requirements. Hamsters and mice showed subtle differences in uterine proliferative and apoptotic patterns during early pregnancy and in response to steroids. There existed almost a direct correlation between apoptosis and caspase-3 expression, suggesting uterine cell death mostly involves the caspase pathway. Consistent with these findings, we showed, for the first time, that execution of uterine epithelial cell apoptosis by caspase-3 is important for blastocyst implantation because a caspsase-3 inhibitor N-acetyl DEVD-CHO when instilled inside the uterine lumen on d 3 of pregnancy inhibits implantation in hamsters and mice. The overall results indicate that uterine cell apoptosis and proliferation patterns are highly ordered cell-specific phenomena that play an important role in maintaining the sexual cycle and pregnancy associated uterine changes. PMID- 16469812 TI - Role of the alternate pathway of dihydrotestosterone formation in virilization of the Wolffian ducts of the tammar wallaby, Macropus eugenii. AB - Dihydrotestosterone in androgen target tissues is formed under most circumstances by the 5alpha-reduction of testosterone, but an alternate pathway involves the oxidation of androstanediol to dihydrotestosterone. To investigate the mechanism by which androgens virilize the Wolffian ducts in the tammar wallaby, [(3)H]progesterone was incubated with testes from d 10 and 19 pouch young, and radioactivity was recovered in testosterone and androstanediol at both ages. Analysis of the intermediates indicates that androstanediol was formed both from testosterone via 5alpha-reduction and 3alpha-keto reduction and directly from 5alpha-reduced progestogens. 5alpha-Reductase activity was high in minces of mesonephros/epididymis from d 6-21 pouch young. When minces of urogenital tract tissues from d 19 pouch young were incubated with [(3)H]testosterone, [(3)H]dihydrotestosterone, and [(3)H]androstanediol, dihydrotestosterone was the principal androgen formed in the mesonephros/epididymis, urogenital sinus, and urogenital tubercle, whereas androstanediol was the principal androgen formed by the testis. In intact pouch young studied between d 10 and 34, administration of the 5alpha-reductase inhibitor, 17beta-(N,N-diethyl)carbamoyl-4-methyl-4-aza 5alpha-androstan-3-one, blocked virilization of the Wolffian ducts in males, and administration of androstanediol caused virilization of the Wolffian ducts in females. We conclude that dihydrotestosterone, largely formed in the tissue by the oxidation of androstanediol derived from the testes and also the 5alpha reduction of testosterone, is responsible for Wolffian duct virilization in this species. PMID- 16469814 TI - Importance of risk stratification models in cardiac surgery. PMID- 16469815 TI - Simultaneous triple kissing stenting in an unprotected left main coronary artery. PMID- 16469813 TI - Indirect regulation of human dehydroepiandrosterone sulfotransferase family 1A member 2 by thyroid hormones. AB - Thyroid hormone, T(3), regulates cell metabolism, differentiation, and development. cDNA microarrays were performed to study the mechanism of target gene regulation after T(3) treatment in a thyroid hormone receptor-alpha (TRalpha)-overexpressing hepatoma cell line (HepG2-TRalpha). The differentially expressed target genes are several metabolic enzymes, including dehydroepiandrosterone-sulfotransferase family 1A member 2 (SULT2A1). Enzyme SULT2A1 was elevated roughly 5-fold at the protein level and 9-fold increase at the mRNA level after 48 h T(3) treatment in HepG2-TRalpha cells. Cycloheximide inhibited T(3)-induced SULT2A1 expression, suggesting that regulation was indirect. SULT2A1 has been reported to be regulated by the two transcription factors, steroidogenic factor 1 (SF1) and GATA, in the human adrenal gland. T(3) induced a 2.5- to 3.5-fold elevation of SF1 at the protein level and a 6.2-fold increase at the RNA level in HepG2-TRalpha cells. About seven SF1 binding sites exist on the SULT2A1 gene. To identify and localize the critical SF1 binding site, series of deletion mutants of SULT2A1 promoter fragments in pGL2 plasmid were constructed. The promoter activity of the SULT2A1 gene was enhanced about 2.8- to 7.1-fold by T(3). The -228 SF1 binding site was identified as the most critical site because deleting this region reduced T(3)-induced expression. Transcription factor SF1 application enhanced the -228 but not -117 reporter plasmid activities. SULT2A1 and SF1 up-regulation at protein and RNA levels in thyroidectomized rats occurred after T(3) application. In summary, this work demonstrated that the SULT2A1 gene was mediated by SF1 and indirectly regulated by T(3). Further study is required to elucidate the physiological importance of SULT2A1 induction mediated by T(3). PMID- 16469816 TI - Alike the metabolic syndrome, hypertension is more than just increased blood pressure. PMID- 16469817 TI - Myotendinous plasticity to ageing and resistance exercise in humans. AB - The age-related loss of muscle mass known as senile sarcopenia is one of the main determinants of frailty in old age. Molecular, cellular, nutritional and hormonal mechanisms are at the basis of sarcopenia and are responsible for a progressive deterioration in skeletal muscle size and function. Both at single-fibre and at whole-muscle level, the loss of force exceeds that predicted by the decrease in muscle size. For single fibres, the loss of intrinsic force is mostly due to a loss in myofibrillar protein content. For whole muscle, in addition to changes in neural drive, alterations in muscle architecture and in tendon mechanical properties, exemplified by a reduction in tendon stiffness, have recently been shown to contribute to this phenomenon. Resistance training can, however, cause substantial gains in muscle mass and strength and provides a protective effect against several of the cellular and molecular changes associated with muscle wasting and weakness. In old age, not only muscles but also tendons are highly responsive to training, since an increase in tendon stiffness has been observed after a period of increased loading. Many of the myotendinous factors characterizing ageing can be at least partly reversed by resistance training. PMID- 16469818 TI - Nitric oxide synthase inhibition reduces O2 cost of force development and spares high-energy phosphates following contractions in pump-perfused rat hindlimb muscles. AB - The purpose of the present experiments was to test the hypotheses that: (i) nitric oxide synthase (NOS) inhibition reduces the O2 cost of force development across a range of contractile demands; and (ii) this reduced O2 cost of force development would be reflected in a sparing of intramuscular higher energy phosphates. Rat distal hindlimb muscles were pump perfused in situ and electrically stimulated (200 ms trains with pulses at 100 Hz, each pulse 0.05 ms duration) for 1 min each at 15, 30 and 60 tetani min(-1) and for 2 min at 90 tetani min(-1) in three groups: 0.01 mM adenosine; 1 mM D-NAME and 0.01 mM adenosine (D-NAME); and 1 mM L-NAME and 0.01 mM adenosine (L-NAME). The gastrocnemius-plantaris-soleus muscle group was freeze clamped post-contractions for metabolite analyses. Force was 19% higher and oxygen uptake (VO2) was 20% lower with L-NAME versus adenosine, and there was a 35% reduction in VO2/time integrated tension versus adenosine and 24% versus D-NAME that was independent of contraction frequency. L-NAME treatment produced a 33% sparing of muscle phosphocreatine (PCr), and intramuscular lactate was no different between groups. In contrast, D-NAME reduced force by 30%, VO2 by 29% and the O2 cost of force development by 15% compared with adenosine, but had no effect on the degree of intramuscular ATP and PCr depletion. These results show that NOS inhibition improved the metabolic efficiency of force development, either by improving the ATP yield for a given O2 consumption or by reducing the ATP cost of force development. In addition, these effects were independent of contraction frequency. PMID- 16469820 TI - Defibrillation of the heart: insights into mechanisms from modelling studies. AB - Despite its critical role in restoring cardiac rhythm and thus in saving human life, cardiac defibrillation remains poorly understood. Further mechanistic inquiry is hampered by the inability of presently available experimental techniques to resolve, with sufficient accuracy, electrical behaviour confined to the depth of the ventricles. The objective of this review article is to demonstrate that realistic 3-D simulations of the ventricular defibrillation process in close conjunction with experimental observations are capable of bringing a new level of understanding of the electrical events that ensue from the interaction between fibrillating myocardium and applied shock. The article does this by reviewing the results of two studies, one on vulnerability to electric shocks and another on defibrillation. An overview of the modelling tools used in these studies is also provided. PMID- 16469819 TI - Trimetazidine improved Ca2+ handling in isoprenaline-mediated myocardial injury of rats. AB - Dysregulation of intracellular Ca2+ homeostasis plays an important role in mediating myocardial injury. We tested the hypothesis that treatment with trimetazidine (TMZ) would improve intracellular Ca2+ handling in myocardial injury of rats. The control group received saline only (10 ml kg(-1) day(-1), i.p.) for 7 days. In a second group, isoprenaline (ISO; 5 mg kg(-1) day(-1), s.c.) was administered to rats for 2 days to induce an acute injury of the myocardium. In a third group, treatment with TMZ (10 mg kg(-1) day(-1), i.p.) was initiated 1 day before ISO administration and continued for 7 days (n = 7 rats in each group). Histopathological evaluation showed that TMZ prevented ISO-induced myocardial damage. TMZ preserved the ATP levels and decreased the maleic dialdehyde (MDA) content in the hearts compared with ISO-treated rats. The diastolic [Ca2+]i measured by loading with fura-2 AM in isolated cardiomyocytes was increased significantly in ISO-treated rats compared to the control animals. TMZ prevented the rise of diastolic [Ca2+]i and the depression of caffeine induced Ca2+ transients caused by ISO administration. The reduction in sarcoplasmic reticulum (SR) Ca2+ content in the heart cells and in cardiac SR Ca2+-ATPase activity in ISO-treated rats was abolished by TMZ, although there were no differences in SR Ca2+-ATPase protein levels between the control, ISO and ISO + 7 mz-treated rats. In addition, TMZ prevented the reduction in sarcolemmal L-type Ca2+ current density in the heart cells induced by ISO treatment. These results demonstrate that the treatment of rats with TMZ inhibited the increase of diastolic [Ca2+]i and prevented the decrease of SR Ca2+ content, SR Ca2+-ATPase activity and L-type Ca2+ current density in cardiomyocytes in ISO-mediated myocardial injury of rats. These changes in Ca2+ handling could help to explain the favourable action of TMZ in myocardial injury. PMID- 16469821 TI - Glucocorticoids promote chondrogenic differentiation of adult human mesenchymal stem cells by enhancing expression of cartilage extracellular matrix genes. AB - In the adult human, mesenchymal stem cells (hMSCs) resident in the bone marrow retain the capacity to proliferate and differentiate along multiple connective tissue lineages, including cartilage. Glucocorticoids (GCs) are required for chondrogenic differentiation of hMSCs in vitro; however, the exact role of GCs in this process is not known. In this study, we examined the effects of dexamethasone (DEX) on chondrogenic differentiation of hMSCs in the presence or absence of DEX, transforming growth factor-beta (TGF-beta), or DEX plus TGF-beta. GC treatment upregulated gene expression of cartilage matrix components aggrecan, dermatopontin, and collagen type XI; enhanced TGF-beta-mediated upregulation of collagen type II and cartilage oligomeric matrix protein; and increased aggrecan and collagen type II production as well as cartilage matrix-sulfated proteoglycans as assessed by immunohistochemistry and alcian blue staining. Inclusion of an antagonist of GCs inhibited expression of chondrogenic differentiation markers, suggesting that the GC effects during chondrogenesis are mediated by the GC receptor (GR). Steady levels of the major active form of GR, GRalpha, were detected in both undifferentiated and differentiating hMSCs, whereas the dominant-negative isoform GRbeta, present at low levels in undifferentiated hMSCs, was downregulated during chondrogenesis. In the presence of DEX and TGF-beta, expression of a collagen type II gene promoter luciferase reporter construct in hMSCs was upregulated. However, coexpression of GRbeta dramatically inhibited promoter activity, suggesting that GRalpha is required for GC-mediated modulation of chondrogenesis and that GCs may play an important role in the maintenance of cartilage homeostasis. PMID- 16469822 TI - Primary prevention of latex related sensitisation and occupational asthma: a systematic review. AB - METHODS: Eight primary prevention intervention studies on natural rubber latex (NRL) published since 1990 were identified and reviewed. This is the largest evidence base of primary prevention studies for any occupational asthmagen. RESULTS: Review of this small and largely observational evidence base supports the following evidence statement: Substitution of powdered latex gloves with low protein powder-free NRL gloves or latex-free gloves greatly reduces NRL aeroallergens, NRL sensitisation, and NRL-asthma in healthcare workers. Evidence in support of this statement is ranked SIGN level 2+, referring to well conducted case-control or cohort studies with a low risk of confounding, bias, or chance and a moderate probability that the relationship is causal. CONCLUSION: Substitution of powdered latex gloves with low protein powder-free NRL gloves or latex-free gloves promises benefits to both workers' health and cost and human resource savings for employers. This message should be broadly disseminated beyond the hospital sector to include other healthcare settings (such as aged care facilities) as well as food service and other industries where latex gloves might be used. PMID- 16469823 TI - The French National Mesothelioma Surveillance Program. AB - OBJECTIVES: The French National Mesothelioma Surveillance Program (NMSP) was established in 1998 by the National Institute for Health Surveillance (InVS). Its objectives are to estimate the trends in mesothelioma incidence and the proportion attributable to occupational asbestos exposure, to help improve its pathology diagnosis, to assess its compensation as an occupational disease, and to contribute to research. METHODS: The NMSP records incident pleural tumours in 21 French districts that cover a population of approximately 16 million people (a quarter of the French population). A standardised procedure of pathological and clinical diagnosis ascertainment is used. Lifetime exposure to asbestos and to other factors (man made mineral fibres, ionising radiation, SV40 virus) is reconstructed, and a case-control study was also conducted. The proportion of mesothelioma compensated as an occupational disease was assessed. RESULTS: Depending on the hypothesis, the estimated number of incident cases in 1998 ranged from 660 to 761 (women: 127 to 146; men: 533 to 615). Among men, the industries with the highest risks of mesothelioma are construction and ship repair, asbestos industry, and manufacture of metal construction materials; the occupations at highest risk are plumbers, pipe-fitters, and sheet-metal workers. The attributable risk fraction for occupational asbestos exposure in men was 83.2% (95% CI 76.8 to 89.6). The initial pathologist's diagnosis was confirmed in 67% of cases, ruled out in 13%, and left uncertain in the others; for half of the latter, the clinical findings supported a mesothelioma diagnosis. In all, 62% applied for designation of an occupational disease, and 91% of these were receiving workers' compensation. CONCLUSIONS: The NMSP is a large scale epidemiological surveillance system with several original aspects, providing important information to improve the knowledge of malignant pleural mesothelioma, such as monitoring the evolution of its incidence, of high risk occupations and economic sectors, and improving pathology techniques. PMID- 16469824 TI - Angiotensin II type 1 receptors and the intestinal microvascular dysfunction induced by ischemia and reperfusion. AB - The acute phase of intestinal ischemia-reperfusion (I/R) injury is mediated by leukocytes and is characterized by oxidative stress and blood cell recruitment. Upregulation of angiotensin II type 1 receptors (AT1-R) has been implicated in the pathogenesis of conditions associated with oxidative stress. The AT1-R antagonist Losartan (Los) attenuates leukocyte recruitment following I/R. However, the role of AT1-R in intestinal I/R injury and the associated platelet leukocyte interactions remains unclear. The objective of this study was to define the contribution of AT1-R to I/R-induced blood cell recruitment in intestinal venules. Leukocyte and platelet adhesion were quantified by intravital microscopy in the small bowel of C57Bl/6 [wild-type (WT)] mice exposed to sham operation or 45 min of ischemia and 4 h of reperfusion. A separate WT group received Los for 7 days before gut I/R (WT-I/R + Los). AT1-R bone marrow chimeras that express AT1-R on the vessel wall but not blood cells also underwent I/R. Platelet and leukocyte adhesion as well as AT1-R expression in the gut microvasculature were significantly elevated after I/R. All of these responses were attenuated in the WT-I/R + Los group, compared with untreated I/R mice. A comparable abrogation of I/R-induced blood cell adhesion was noted in AT1-R bone marrow chimeras. I/R induced platelet adhesion was unaltered in mice overexpressing Cu,Zn-SOD or mice deficient in NAD(P)H oxidase. These data suggest that although gut I/R upregulates endothelial expression of AT1-R, engagement of these angiotensin II receptors on blood cells is more important in eliciting the prothrombogenic and proinflammatory state observed in postischemic gut venules, through a superoxide independent pathway. PMID- 16469825 TI - Ghrelin acts on the dorsal vagal complex to stimulate pancreatic protein secretion. AB - Ghrelin receptors are present in the central nervous system. We hypothesized that ghrelin released from the stomach acts as an endocrine substance and stimulates brain stem vagovagal circuitry to evoke pancreatic secretion. In an in vivo anesthetized rat model, an intravenous infusion of ghrelin at doses of 5, 10, and 25 nmol increased pancreatic protein secretion from a basal level of 125 +/- 6 to 186 +/- 8, 295 +/- 12, and 356 +/- 11 mg/h, respectively. Pretreatment with atropine or hexamethonium or an acute vagotomy, but not a perivagal application of capsaicin, completely abolished pancreatic protein secretion responses to ghrelin. In conscious rats, an intravenous infusion of ghrelin at a dose of 10 nmol resulted in a 2.2-fold increase in pancreatic protein secretion over basal volume. Selective ablation of the area postrema abolished pancreatic protein secretion stimulated by intravenous infusion of ghrelin but did not alter the increase in pancreatic protein secretion evoked by diversion of bile-pancreatic juice. Immunohistochemical staining showed a marked increase in the number of c Fos-expressing neurons in the area postrema, nucleus of the solitary tract, and dorsal motor nucleus of the vagus after an intravenous infusion of ghrelin in sham-lesioned rats; selective ablation of the area postrema eliminated this increase. In conclusion, ghrelin stimulates pancreatic secretion via a vagal cholinergic efferent pathway. Circulating ghrelin gains access to the brain stem vagovagal circuitry via the area postrema, which represents the primary target on which peripheral ghrelin may act as an endocrine substance to stimulate pancreatic secretion. PMID- 16469826 TI - Short-chain fatty acid mediated phosphorylation of heat shock protein 25: effects on camptothecin-induced apoptosis. AB - Although short-chain fatty acid (SCFA)-induced heat shock protein 25 (Hsp25) is associated with increased cellular resistance to injury, withdrawal of lumenal butyrate in vivo is associated with intestinal epithelial injury and apoptosis. Recognizing that SCFA-dependent posttranslational modification of Hsp25 may involve altered Hsp25 phosphorylation, we hypothesized that butyrate regulates Hsp25 phosphorylation and secondarily affects cellular responses to apoptosis inducing agents. Intestinal epithelial crypt IEC-18 cells were treated with butyrate, propionate, or the histone deacetylase inhibitor trichostatin A for 6 24 h. Immunolocalization of Hsp25 was examined by confocal laser microscopy. Hsp25 phosphorylation was characterized using two-dimensional isoelectric focusing gel electrophoresis. Hsp25 accumulation in cytoskeletal- and mitochondrial-enriched fractions was examined by immunoblotting. The activation of p38 MAP kinase was determined using phospho-specific antibodies and MAPKAPK 2 kinase assays. The effects of SCFA on apoptosis were studied by ELISA detection of cleaved DNA and using antibodies recognizing cleaved caspase-3. Five millimolar butyrate induced no significant injury to IEC-18 cells. Hsp25 did not accumulate in Triton X-100-insoluble cytoskeletal fractions with butyrate treatment but did localize to mitochondria in a p38 MAP kinase-dependent manner. Hsp25 phosphorylation was induced by butyrate, propionate, and trichostatin A. Butyrate-mediated changes in Hsp25 phosphorylation coincide with the activation of the p38 MAP kinase and MAPKAPK 2. Butyrate, propionate, and low-dose trichostatin A confer significant protection from camptothecin-induced apoptosis, which was not reversed by the p38 inhibitor SB203580. We conclude that butyrate mediated phosphorylation of Hsp25 is associated with significant resistance to apoptosis, which appears to be independent of p38-mediated targeting of Hsp25 to mitochondria. PMID- 16469827 TI - Differential regulation of the JNK/AP-1 pathway by S-adenosylmethionine and methylthioadenosine in primary rat hepatocytes versus HuH7 hepatoma cells. AB - S-adenosylmethionine (AdoMet) and 5'-methylthioadenosine (MTA) exert a protective action on apoptosis induced by okadaic acid in primary rat hepatocytes but not in human transformed HuH7 cells. In the present work, we analyzed the role played by the JNK/activator protein (AP)-1 pathway in this differential effect. Okadaic acid induced the phosphorylation of JNK and c-Jun and the binding activity of AP 1 in primary hepatocytes, and pretreatment with either AdoMet or MTA prevented those effects. In HuH7 cells, pretreatment with either AdoMet or MTA did not affect JNK and c-Jun activation or AP-1 binding induced by okadaic acid. In both cell types, p38 was activated by okadaic acid, but neither AdoMet nor MTA presented a significant effect on its activity. Therefore, the differential effect of both AdoMet and MTA on the JNK/AP-1 pathway could explain their antiapoptotic effect on primary hepatocytes and the lack of protection they show against okadaic acid-induced apoptosis in hepatoma cells. PMID- 16469828 TI - Helicobacter pylori-secreted factors inhibit dendritic cell IL-12 secretion: a mechanism of ineffective host defense. AB - Helicobacter pylori evades host immune defenses and causes chronic gastritis. Immunity against intestinal pathogens is largely mediated by dendritic cells, yet the role of dendritic cells in acute H. pylori infection is largely unknown. We observed the recruitment of dendritic cells to the gastric mucosa of H. pylori infected mice. Bone marrow-derived dendritic cells from mice responded to live H. pylori by upregulating the expression of proinflammatory cytokine mRNA (i.e., IL 1alpha, IL-1beta, and IL-6). The supernatant from dendritic cells stimulated with H. pylori for 18 h contained twofold higher levels of IL-12p70 than IL-10 and induced the proliferation of syngeneic splenocytes and type 1 T helper cell cytokine release (IFN-gamma and TNF-alpha). These responses were significantly lower compared with those induced by Acinetobacter lwoffi, another gastritis causing pathogen more susceptible to host defenses. Analysis of whole H. pylori sonicate revealed the presence of a heat-stable factor secreted from H. pylori that specifically inhibited IL-12 but not IL-10 release from dendritic cells activated by A. lwoffi. Our findings suggest that dendritic cells participate in the host immune response against H. pylori and that their suppression by H. pylori may explain why infected hosts fail to prevent bacterial colonization. PMID- 16469829 TI - Foxl1-deficient mice exhibit aberrant epithelial cell positioning resulting from dysregulated EphB/EphrinB expression in the small intestine. AB - The winged helix transcription factor Foxl1, expressed in the gut mesenchyme, regulates epithelial cell proliferation and differentiation through the Wnt/beta catenin pathway. To better understand the role of Foxl1 in epithelial morphogenesis, we examined the tissue structure and positioning of epithelial cells in the small intestine of Foxl1-deficient mice. The small intestine of Foxl1-deficient mice manifested aberrant crypt structure, including widely distributed Paneth cells, which coincided with the ectopic and increased expression of EphB2 and EphB3, which are key regulators of epithelial cell positioning. Furthermore, real-time quantitative PCR indicated that a subset of Wnt family genes was highly expressed in the gut mesenchyme of Foxl1-deficient mice compared with that of wild-type mice. Such an increase in Wnt expression was remarkable in the mesenchyme, where the aberrant Paneth cell positioning was observed by in situ hybridization. Foxl1 plays an important role in the maintenance of crypt architecture and epithelial cell positioning through the mesenchymal-epithelial interaction in the small intestine. This interaction is essential for the normal regulation of the Wnt/beta-catenin pathway and the subsequent EphB/EphrinB expression. PMID- 16469830 TI - Development of fetal haemoglobin-blood cells (F cells) within colorectal tumour tissues. AB - AIM: To evaluate the sources of fetal haemoglobin (HbF) as an indicator in cancer. An immunohistochemical study was carried out on some of the most common kinds of cancer. All of these cancers had serologically high levels of HbF as evaluated previously. METHODS: Immunoaffinity-purified anti-HbF was immunohistochemically used to study F cell distribution in the following cancers: colorectal adenocarcinoma, urinary bladder transitional cell carcinoma, brain tumours, lung carcinoma, breast adenocarcinoma, leukaemia, Burkitt's lymphoma and endometrial carcinoma. RESULTS: In colorectal adenocarcinoma, HbF-containing red blood cells (FRBC) were present within thin-walled vessels or were disposed in dense clusters within the tumour. Some of these cells were nucleated or binucleated HbF-erythroblasts or HbF-normoblasts (FNBS). In two cases, high levels of mitoses within HbF-erythroblasts were observed. In half of the cases with transitional cell carcinoma of the urinary bladder, regional intratumoral blood vessels were found to contain 5-50% FRBC. In the other tumours examined, F cells were not observed. FRBCs, however, were occasionally observed in the regional lymph nodes of some of these cancers. CONCLUSIONS: The evaluation of HbF as a potential plasma marker is suggested by the high concentration of FRBCs in colorectal tumours. The apparent development of FRBCs in colorectal tumour tissues is an interesting observation, as these cells were previously thought to develop in medullary or lymphoid tissues. It is thus suggested that the colonic microenvironment may stimulate extramedullary fetal-type haematopoiesis. PMID- 16469831 TI - Metabolomic profiling of heat stress: hardening and recovery of homeostasis in Drosophila. AB - Frequent exposure of terrestrial insects to temperature variation has led to the evolution of protective biochemical and physiological mechanisms, such as the heat shock response, which markedly increases the tolerance to heat stress. Insight into such mechanisms has, so far, mainly relied on selective studies of specific compounds or characteristics or studies at the genomic or proteomic levels. In the present study, we have used untargeted NMR metabolomic profiling to examine the biological response to heat stress in Drosophila melanogaster. The metabolite profile was analyzed during recovery after exposure to different thermal stress treatments and compared with untreated controls. Both moderate and severe heat stress gave clear effects on the metabolite profiles. The profiles clearly demonstrated that hardening by moderate heat stress led to a faster reestablishment of metabolite homeostasis after subsequent heat stress. Several metabolites were identified as responsive to heat stress and could be related to known physiological and biochemical responses. The time course of the recovery of metabolite homeostasis mirrored general changes in gene expression, showing that recovery follows the same temporal pattern at these two biological levels. Finally, our data show that heat hardening permits a quicker return to homeostasis, rather than a reduction of the acute metabolic perturbation and that the reestablishment of homeostasis is important for obtaining maximal heat hardening effect. The results display the power of NMR metabolomic profiling for characterization of the instantaneous physiological condition, enabling direct visualization of the perturbation of and return to homeostasis. PMID- 16469832 TI - Hypothermia-enhanced splenic cytokine gene expression is independent of the sympathetic nervous system. AB - Splenic nerve denervation abrogates enhanced splenic cytokine gene expression responses to acute heating, demonstrating that hyperthermia-induced activation of splenic sympathetic nerve discharge (SND) increases splenic cytokine gene expression. Hypothermia alters SND responses; however, the role of the sympathetic nervous system in mediating splenic cytokine gene expression responses to hypothermia is not known. The purpose of the present study was to determine the effect of hypothermia on the relationship between the sympathetic nervous system and splenic cytokine gene expression in anesthetized F344 rats. Gene expression analysis was performed using a microarray containing 112 genes, representing inflammatory cytokines, chemokines, cytokine/chemokine receptors and housekeeping genes. A subset of differentially expressed genes was verified by real-time RT-PCR analysis. Splenic SND was decreased significantly during cooling (core temperature decreased from 38 to 30 degrees C) in splenic-intact rats but remained unchanged in sham-cooled splenic-intact rats (core temperature maintained at 38 degrees C). Hypothermia upregulated the transcripts of several genes, including, chemokine ligands CCL2, CXCL2, CXCL10, and CCL20, and interleukins IL-1alpha, IL-1beta, and IL-6. Gene expression responses to hypothermia were similar for the majority of cytokine genes in splenic-intact and splenic-denervated rats. These results suggest that hypothermia-enhanced splenic cytokine gene expression is independent of splenic SND. PMID- 16469833 TI - Testosterone and estrogen have opposing actions on inflammation-induced plasma extravasation in the rat temporomandibular joint. AB - The present study was designed to test the hypothesis that estrogen exacerbates inflammation of the temporomandibular joint (TMJ). Evans blue dye was used to quantify plasma extravasation (PE) around the rat TMJ. In an initial set of experiments, TMJ PE was compared in naive intact male and female rats, as well as in both groups after complete Freund's adjuvant (CFA)-induced inflammation of the TMJ. In contrast to our hypothesis, TMJ PE was significantly greater in both naive and CFA-inflamed male rats than in females. To determine whether these differences were due to gonadal hormones, four additional groups of rats were studied: gonadectomized (Gx) males and females, Gx males with chronic testosterone (T) replacement, and Gx females with chronic estrogen (E) replacement. The sex difference in baseline TMJ PE appeared to reflect the actions of T. However, in the presence of TMJ inflammation, T augmented TMJ PE in males, while E attenuated TMJ PE in females. Changes in PE were also assessed in the contralateral TMJ. Results from this analysis indicated that there is a transient contralateral increase in TMJ PE in females but not males. Given that there is an inverse relationship between PE and joint damage, our results suggest that testosterone may mitigate, but estrogen may exacerbate, TMJ damage, particularly in the presence of overt inflammation. Importantly, our results may help explain both the higher prevalence and severity of temporomandibular disorder pain in females than males. PMID- 16469834 TI - Visually guided whole cell patch clamp of mouse supraoptic nucleus neurons in cultured and acute conditions. AB - Recent advances in neuronal culturing techniques have supplied a new set of tools for studying neural tissue, providing effective means to study molecular aspects of regulatory elements in the supraoptic nucleus of the hypothalamus (SON). To combine molecular biology techniques with electrophysiological recording, we modified an organotypic culture protocol to permit transfection and whole cell patch-clamp recordings from SON cells. Neonatal mouse brain coronal sections containing the SON were dissected out, placed on a filter insert in culture medium, and incubated for at least 4 days to allow attachment to the insert. The SON was identifiable using gross anatomical landmarks, which remained intact throughout the culturing period. Immunohistochemical staining identified both vasopressinergic and oxytocinergic cells present in the cultures, typically appearing in well-defined clusters. Whole cell recordings from these cultures demonstrated that certain properties of the neonatal mouse SON were comparable to adult mouse magnocellular neurons. SON neurons in both neonatal cultures and acute adult slices showed similar sustained outward rectification above -60 mV and action potential broadening during evoked activity. Membrane potential, input resistance, and rapidly inactivating potassium current density (IA) were reduced in the cultures, whereas whole cell capacitance and spontaneous synaptic excitation were increased, perhaps reflecting developmental changes in cell physiology that warrant further study. The use of the outlined organotypic culturing procedures will allow the study of such electrophysiological properties of mouse SON using whole cell patch-clamp, in addition to various molecular, techniques that require longer incubation times. PMID- 16469835 TI - Intranuclear coupling of hypothalamic magnocellular nuclei by glutamate synaptic circuits. AB - Magnocellular neurons of the supraoptic nucleus (SON) and paraventricular nucleus (PVN) display bursting activity that is synchronized under certain conditions. They receive excitatory synaptic inputs from intrahypothalamic glutamate circuits, some of which are activated by norepinephrine. Ascending noradrenergic afferents and intrahypothalamic glutamate circuits may be responsible for the generation of synchronous bursting among oxytocin neurons and/or asynchronous bursting among vasopressin neurons located in the bilateral supraoptic and paraventricular nuclei. Here, we tested whether magnocellular neurons of the PVN receive excitatory synaptic input from the contralateral PVN and the region of the retrochiasmatic SON (SONrx) via norepinephrine-sensitive internuclear glutamate circuits. Whole cell patch-clamp recordings were performed in PVN magnocellular neurons in coronal hypothalamic slices from male rats, and the ipsilateral SONrx region and contralateral PVN were stimulated using electrical and chemical stimulation. Electrical and glutamate microdrop stimulation of the ipsilateral SONrx region or contralateral PVN elicited excitatory postsynaptic potentials/currents (EPSP/Cs) in PVN magnocellular neurons mediated by glutamate release, revealing internuclear glutamatergic circuits. Microdrop application of norepinephrine also elicited EPSP/Cs, suggesting that these circuits could be activated by activation of noradrenergic receptors. Repetitive electrical stimulation and drop application of norepinephrine, in some cases, elicited bursts of action potentials. Our data reveal glutamatergic synaptic circuits that interconnect the magnocellular nuclei and that can be activated by norepinephrine. These internuclear glutamatergic circuits may provide the functional architecture to support burst generation and/or burst synchronization in hypothalamic magnocellular neurons under conditions of activation. PMID- 16469836 TI - Roles of glutamatergic and serotonergic mechanisms in reflex control of the external urethral sphincter in urethane-anesthetized female rats. AB - This study was conducted to examine reflex mechanisms that mediate urinary bladder and external urethral sphincter (EUS) coordination in urethane anesthetized female Sprague-Dawley rats. We investigated the properties of EUS reflexes elicited by electrical stimulation of pelvic nerve afferent axons (pelvic-EUS reflex). The changes in the reflexes induced by bladder distension and administration of agonists or antagonists for glutamatergic or serotonergic receptors were examined. The reflexes consisted of an early response (ER, 18- to 22-ms latency) and a late, long-duration (>100-ms latency) response (LR), which consisted of bursts of activity at 20- to 160-ms interburst intervals. In a few experiments, a reflex with an intermediate (40- to 70-ms) latency was also identified. With the bladder empty, the ER, but not the LR, was detected in the majority of experiments. The LR was markedly enhanced when the bladder was distended. The ER remained, but the LR was abolished, after spinal cord transection at T8-T9. The ER and LR were significantly decreased 75 and 35%, respectively, by the N-methyl-D-aspartate receptor antagonist MK-801 (0.3 mg/kg iv), but only decreased 18 and 14%, respectively, by the alpha-amino-5 methylisoxazole-4-propionate receptor antagonist LY-215490 (3 mg/kg iv). The serotonin (5-HT1A) receptor agonist 8-hydroxy-2-(di-n-propylamino)-tetralin (1 mg/kg iv) enhanced spontaneous EUS activity and the pelvic-EUS reflex. WAY-100635 (0.1-1 mg/kg iv), a 5-HT1A antagonist, reversed the effect of 8-hydroxy-2-(di-n propylamino)-tetralin and suppressed EUS activity and the pelvic-EUS reflex. These results indicate that glutamatergic and serotonergic mechanisms are important in the reflex pathways underlying bladder- sphincter coordination in rats. PMID- 16469839 TI - A novel approach to assess insulin sensitivity reveals no increased insulin sensitivity in mice with a dominant-negative mutant hepatocyte nuclear factor 1alpha. AB - In phenotype experiments in mice, determination of dynamic insulin sensitivity often uses the insulin tolerance test. However, the interpretation of this test is complicated by the counterregulation occurring at low glucose. To overcome this problem, we determined the dynamic insulin sensitivity after inhibition of endogenous insulin secretion by diazoxide (25 mg/kg) in association with intravenous administration of glucose plus insulin (the DSGIT technique). Estimation of insulin sensitivity index (SI) by this technique showed good correlation to SI from a regular intravenous glucose tolerance test (r = 0.87; P < 0.001; n = 15). With DSGIT, we evaluated dynamic insulin sensitivity in mice with a rat insulin promoter (beta-cell-targeted) dominant-negative mutation of hepatic nuclear factor (HNF)-1alpha [RIP-DN HNF-1alpha (Tg) mice]. When insulin was administered exogenously at the same dose in Tg and wild-type (WT) mice, plasma insulin levels were higher in WT, indicating an increased insulin clearance in Tg mice. When the diazoxide test was used, different doses of insulin were therefore administered (0.1 and 0.15 U/kg in WT and 0.2 and 0.25 U/kg in Tg) to achieve similar insulin levels in the groups. Minimal model analysis showed that SI was the same in the two groups (0.78 +/- 0.21 x 10(-4) min.pmol(-1).l(-1) in WT vs. 0.60 +/- 0.11 in Tg; P = 0.45) as was the glucose elimination rate (P = 0.27). We conclude that 1) the DSGIT technique determines the in vivo dynamic insulin action in mice, 2) insulin clearance is increased in Tg mice, and 3) chronic islet dysfunction in RIP-DN HNF-1alpha mice is not compensated with increased insulin sensitivity. PMID- 16469838 TI - Regulation of plasma vasopressin and renin activity in conscious hindlimb unloaded rats. AB - Cardiovascular deconditioning occurs in astronauts after spaceflight or in individuals subjected to bed rest. It is characterized by an increased incidence of orthostatic intolerance. The mechanisms responsible for orthostatic intolerance are likely multifactorial and may include hypovolemia, autonomic dysfunction, and vascular and cardiac alterations. The arterial baroreflex is an important compensatory mechanism in the response to an orthostatic stress. In a previous study, we demonstrated that arterial baroreflex mediated sympathoexcitation was blunted in hindlimb-unloaded (HU) rats, a model of cardiovascular deconditioning. The arterial baroreflex also contributes to the regulation of vasoactive hormones including vasopressin and angiotensin II. In the present study, we tested the hypothesis that the neurohumoral response to hypotension is also attenuated in rats after 14 days of hindlimb unloading. To test this hypothesis, the vasodilator diazoxide (15 or 25 mg/kg) or saline (0.9%) was administered to produce hypotension or control conditions, respectively, in conscious HU and control rats. Plasma samples were collected and assayed for vasopressin and plasma renin activity (PRA). Diazoxide (25 mg/kg) produced significant increases in vasopressin and PRA compared with saline controls. HU rats exhibited significantly higher levels of vasopressin at rest and the increase in vasopressin levels during hypotension was enhanced by hindlimb unloading. Neither resting nor hypotension-induced PRA was altered by hindlimb unloading. These data suggest that although baroreflex-mediated sympathoexcitation is blunted by hindlimb unloading, hypotension-induced vasopressin release is enhanced and hypotension-induced PRA is unaffected. Increased circulating vasopressin may serve to compensate for blunted baroreflex regulation of sympathetic nervous activity produced by hindlimb unloading or may actually contribute to it. PMID- 16469837 TI - Effects of prenatal ethanol exposure and sex on the arginine vasopressin response to hemorrhage in the rat. AB - AVP synthesis, storage, and osmotically stimulated release are reduced in young adult rats exposed prenatally to ethanol (PE). Whether the reduced release of AVP to the osmotic stimulus is due to impairment of the vasopressin system or specifically to an osmoreceptor-mediated release is not known. The present experiments were done, therefore, to determine whether a hemorrhage-induced AVP response would also be diminished in PE-exposed rats. Pregnant rats were fed either a control liquid diet [no prenatal ethanol (NPE)] or a liquid diet with 35% of the calories from ethanol from days 7-21 of pregnancy. Offspring were weaned at 3 wk of life. At 11 wk of age, femoral arterial catheters were surgically placed, and blood volumes were determined at 12 wk. Three days later, two hemorrhages of 10% of the blood volume were performed with samples taken before and 10 min after the hemorrhages. After a 20% blood loss, plasma AVP was 19% higher in NPE rats than in the PE rats despite no differences in mean arterial blood pressure (MABP). Also, hypothalamic AVP mRNA and pituitary AVP content were reduced in PE rats. Furthermore, confirming an earlier report of sex differences in AVP release, the hemorrhage-induced hormone response was twofold greater in female rats than male rats, regardless of previous ethanol exposure. These studies demonstrate that the AVP response to hemorrhage is reduced in PE rats independently of differences in MABP. The data are compatible with a theory of a reduced number of hemorrhage-responsive vasopressinergic neurons capable of stimulated AVP release in PE rats. PMID- 16469840 TI - Androgens augment renal vascular responses to ANG II in New Zealand genetically hypertensive rats. AB - Males develop higher blood pressure than do females. This study tested the hypothesis that androgens enhance responsiveness to ANG II during the development of hypertension in New Zealand genetically hypertensive (NZGH) rats. Male NZGH rats were obtained at 5 wk of age and subjected to sham operation (Sham) or castration (Cas) then studied at three age groups: 6-7, 11-12, and 16-17 wk. Mean arterial blood pressure (MAP), heart rate (HR), and renal blood flow (RBF) measurements were recorded under Inactin anesthesia. These variables were measured after enalapril (1 mg/kg) treatment and during intravenous ANG II infusion (20, 40, and 80 ng/kg/min). Plasma testosterone was measured by ELISA. Angiotensin type 1 (AT1) receptor expression was assessed by Western blot analysis and RT-PCR. ANG II-induced MAP responses were significantly attenuated in Cas NZGH rats. At the highest ANG II dose, MAP increased by 40+/-4% in Sham vs. 22+/-1% in Cas NZGH rats of 16-17 wk of age. Similarly, renal vascular resistance (RVR) responses to ANG II were reduced by castration (209+/-20% in Sham vs. 168+/-10% in Cas NZGH rats at 16-17 wk of age). Castration also reduced MAP recorded in conscious NZGH rats of this age group. Testosterone replacement restored baseline MAP and the pressor and RVR responses to ANG II. Castration reduced testosterone concentrations markedly. Testosterone treatment restored these concentrations. Neither castration nor castration+testosterone treatment affected AT1 receptor mRNA or protein expression. Collectively, these data suggest that androgens modulate renal and systemic vascular responsiveness to ANG II, which may contribute to androgen-induced facilitation of NZGH rat hypertension. PMID- 16469841 TI - Cadmium and calcium uptake in isolated mitochondria-rich cell populations from the gills of the freshwater rainbow trout. AB - A novel cell isolation technique was used to characterize cadmium and calcium uptake in distinct populations of gill cells from the adult rainbow trout (Oncorhynchus mykiss). A specific population of mitochondria-rich (MR) cell, termed the PNA+ MR cell (PNA is peanut lectin agglutinin), was found to accumulate over threefold more 109Cd than did PNA- MR cells, pavement cells (PV cells), and mucous cells during a 1-h in vivo exposure at 2.4 microg/l 109Cd. In vitro 109Cd exposures, performed in standard PBS and Cl- -free PBS, at concentrations from 1 to 16 microg/l 109Cd, were also carried out to further characterize Cd2+ uptake kinetics. As observed during in vivo experiments, PNA+ MR cells accumulated significantly more 109Cd than did other cell types when exposures were performed by an in vitro procedure in PBS. Under such conditions, Cd2+ accumulation kinetics in all cell types could be described with Michaelis Menten relationships, with Km values of approximately 3.0 microg/l Cd (27 nM) for both MR cell subtypes and 8.6 microg/l Cd (77 nM) for PV cells. In similar experiments performed in Cl- -free conditions, a significant reduction in 109Cd accumulation in PNA+ MR cells was seen but not in PNA- MR or in PV cells. In vitro 45Ca fluxes were also performed to determine the cellular localization of Ca2+ transport in these functionally distinct populations of gill cells. 45Ca uptake was most pronounced in PNA+ MR cells, with levels over threefold higher than those found in either PNA(-) MR or in PV cells. Results from the present study suggest that the PNA+ MR cell type is a high-affinity and high-capacity site for apical entry of Cd2+ and Ca2+ in the gill epithelium of rainbow trout. PMID- 16469842 TI - Mitochondrial carriers in the cytoplasmic state have a common substrate binding site. AB - Mitochondrial carriers link biochemical pathways in the cytosol and mitochondrial matrix by transporting substrates across the inner mitochondrial membrane. Substrate recognition is specific for each carrier, but sequence similarities suggest the carriers have similar structures and mechanisms of substrate translocation. By considering conservation of amino acids, distance and chemical constraints, and by modeling family members on the known structure of the ADP/ATP translocase, we have identified a common substrate binding site. It explains substrate selectivity and proton coupling and provides a mechanistic link to carrier opening by substrate-induced perturbation of the salt bridges that seal the pathway to and from the mitochondrial matrix. It enables the substrate specificity of uncharacterized mitochondrial carriers to be predicted. PMID- 16469843 TI - Principles of self-assembly of helical pores from dendritic dipeptides. AB - The self-assembly of the dendritic dipeptides (4-3,4-3,5)nG2-CH2-Boc-L-Tyr-L-Ala OMe and their achiral dendritic alcohol (4-3,4-3,5)nG2-CH2OH precursors, both with n = 1-16, where n represents the number of methylenic units in the alkyl groups of the dendron, are reported. All chiral dendritic dipeptides and achiral dendritic alcohols self-assemble into helical porous columns that are stable in both solution and solid state. The pore diameter (D(pore)) of the columns self assembled from dendritic dipeptides is approximately 10 A larger than that of structures assembled from dendritic alcohols. The increase of the D(pore) at the transition from dendritic alcohol to dendritic dipeptide is accompanied by a decreased solid angle of the building block. This trend is in agreement with previous pore size-solid angle dependences observed with different protective groups of the dipeptide and primary structures of the dendron. However, within the series of dendritic alcohols and dendritic dipeptides with various n, the D(pore) increases when the solid angle increases. The results of these investigations together with those of previous studies on the role of dipeptide stereochemistry and protective groups on this self-assembly process provide the molecular principles required to program the construction of supramolecular helical pores with diameter controlled at the A level from a single dendritic dipeptide architecture. These principles are expected to be valid for libraries of dendritic dipeptides based on dendrons and dipeptides with various primary structures. PMID- 16469844 TI - Hypoxia-inducible myoglobin expression in nonmuscle tissues. AB - Myoglobin (Myg) is an oxygen-binding hemoprotein that is widely thought to be expressed exclusively in oxidative skeletal and cardiac myocytes, where it plays a key role in coping with chronic hypoxia. We now show in a hypoxia-tolerant fish model, that Myg is also expressed in a range of other tissues, including liver, gill, and brain. Moreover, expression of Myg transcript was substantially enhanced during chronic hypoxia, the fold-change induction being far greater in liver than muscle. By using 2D gel electrophoresis, we have confirmed that liver expresses a protein corresponding to the Myg-1 transcript and that it is significantly up-regulated during hypoxia. We have also discovered a second, unique Myg isoform, distinct from neuroglobin, which is expressed exclusively in the neural tissue but whose transcript expression was unaffected by environmental hypoxia. Both observations of nonmuscle expression and a brain-specific isoform are unprecedented, indicating that Myg may play a much wider role than previously understood and that Myg might function in the protection of tissues from deep hypoxia and ischemia as well as in reoxygenation and reperfusion injury. PMID- 16469845 TI - Homotypic fusion of early endosomes: SNAREs do not determine fusion specificity. AB - Membrane fusion in the secretory pathway is mediated by soluble N-ethylmaleimide sensitive factor attachment receptor (SNARE) proteins. Different fusion steps are thought to be effected by independent sets of SNAREs, but it is unclear whether specificity is determined by an intrinsic specificity of SNARE pairing or by upstream factors. Using a newly developed microscopy-based assay, we have investigated the SNARE specificity of homotypic early endosomal fusion. We show that early endosomes contain multiple sets of SNAREs, including, in addition to the putative early endosomal SNAREs, those involved in exocytosis and in fusion of late endosomes. We demonstrate that fusion is largely mediated by a complex formed by syntaxin 13, syntaxin 6, vti1a, and VAMP4, whereas the exocytic and late endosomal SNAREs play little or no role in the reaction. In contrast, proteoliposomes reconstituted with early endosomal SNAREs promiscuously fuse with liposomes containing exocytotic or late endosomal SNAREs. We conclude that the specificity of SNARE pairing does not suffice to determine the specificity of organelle fusion. PMID- 16469846 TI - Docosahexaenoic acid selectively inhibits plasma membrane targeting of lipidated proteins. AB - Membrane localization of lipidated cytosolic signaling proteins is mediated by interactions between specific lipid anchors and membranes, but little is known about the regulatory role of membrane composition in lipidated protein membrane targeting. Here, using green fluorescent protein (GFP) chimeras and quantitative fluorescence microscopy in living mouse colonocytes, we show that docosahexaenoic acid (DHA), a dietary polyunsaturated fatty acid (PUFA) with membrane lipid modifying properties, selectively inhibits plasma membrane (PM) targeting and increases the endomembrane localization of lipidated proteins that are cytoplasmic cargo in the exocytic pathway, without affecting the exocytic pathway itself. DHA selectivity seems to be dictated by the protein trafficking route, independent of the functional state of proteins and the location and composition of membrane anchors. DHA enrichment in cell membranes was required to elicit the inhibitory effect. These data reveal that membrane lipid composition influences cell signaling by modulating intracellular trafficking and localization of membrane proteins, providing a potential molecular mechanism for the documented health benefits of DHA. PMID- 16469847 TI - Transcription factor gata4 regulates cardiac BCL2 gene expression in vitro and in vivo. AB - The transcription factor GATA-4 protects cardiomyocytes against doxorubicin induced cardiotoxicity. Here, we report the identification of Bcl2 as a direct target gene of GATA4 that may mediate the prosurvival function of GATA4 in cardiomyocytes. Bcl2 transcript and protein levels were reduced by doxorubicin in neonatal rat ventricular cardiomyocytes (NRVC) and in mouse heart as determined by RT-PCR and Western blot analysis. The reduction in Bcl2 was prevented by overexpression of GATA4 in NRVC and in transgenic mouse heart. Also, expression of GATA4 increased baseline Bcl2 levels by 30% in NRVC and 2.7-fold in transgenic heart, indicating the sufficiency of GATA4 to up-regulate Bcl2 gene expression. GATA4 knockdown by siRNA reduced Bcl2 levels by 48% in NRVC, suggesting that GATA4 is required for Bcl2 constitutive gene expression. Transfection of HEK cells with GATA4 plasmids activated Bcl2 promoter and elevated Bcl2 protein levels. Deletion and mutagenesis analysis revealed that a consensus GATA motif at base -266 on the promoter conserved across multiple species is partially responsible for the promoter activity. Electrophoretic mobility shift and chromatin immunoprecipitation assays demonstrate that GATA4 directly bound to this GATA site. Together, these results indicate that GATA4 positively regulates cardiac Bcl2 gene expression in vitro and in vivo. PMID- 16469849 TI - Biosurfactants: potential applications in medicine. AB - The use and potential commercial application of biosurfactants in the medical field has increased during the past decade. Their antibacterial, antifungal and antiviral activities make them relevant molecules for applications in combating many diseases and as therapeutic agents. In addition, their role as anti-adhesive agents against several pathogens indicates their utility as suitable anti adhesive coating agents for medical insertional materials leading to a reduction in a large number of hospital infections without the use of synthetic drugs and chemicals. This review looks at medicinal and therapeutic perspectives on biosurfactant applications. PMID- 16469850 TI - Successful voriconazole therapy of disseminated Fusarium verticillioides infection in an immunocompromised patient receiving chemotherapy. PMID- 16469848 TI - The triage of damaged proteins: degradation by the ubiquitin-proteasome pathway or repair by molecular chaperones. AB - Accumulation of damaged proteins is causally related to many age-related diseases. The ubiquitin-proteasome pathway (UPP) plays a role in selective degradation of damaged proteins, whereas molecular chaperones, such as heat shock proteins, are involved in refolding denatured proteins. This work demonstrates for the first time that the UPP and molecular chaperones work in a competitive manner and that the fates of denatured proteins are determined by the relative activities of the UPP and molecular chaperones. Enhanced UPP activity suppresses the refolding of denatured proteins whereas elevated chaperone activity inhibits the degradation of denatured proteins. CHIP, a co-chaperone with E3 activity, plays a pivotal role in determining the fates of the damaged proteins. The delicate balance between UPP-mediated degradation and refolding of denatured proteins is governed by relative levels of CHIP and other molecular chaperones. Isopeptidases, the enzymes that reverse the actions of CHIP, also play an important role in determining the fate of denatured proteins. PMID- 16469851 TI - Macrolide resistance of Staphylococcus aureus and Haemophilus species associated with long-term azithromycin use in cystic fibrosis. AB - OBJECTIVES: Azithromycin is used to modulate exuberant inflammatory response in patients with cystic fibrosis (CF). The purpose of this study was to determine the association between long-term use of azithromycin in CF patients and change over time in macrolide susceptibility of Staphylococcus aureus and Haemophilus spp. METHODS: The study was performed at the Erasmus MC-Sophia Children's Hospital. CF patients' sputum cultures were obtained at routine visits and at pulmonary exacerbations. All cultures between January 1999 and March 2004 were included. Antibiotic susceptibility of S. aureus and Haemophilus spp. was tested routinely. Susceptibility was compared with isolates from sputum of non-CF patients. Logistic regression was used to analyse the association between azithromycin use and resistance, adjusting for age, Pseudomonas carriage and time trends. RESULTS: In March 2004 one-third of CF patients were on azithromycin maintenance treatment. S. aureus (715 isolates) and/or Haemophilus (537 isolates) were cultured in 141 of the 155 patients on one or more occasions. The study period was divided into octiles. Erythromycin resistance in S. aureus increased from 6.9 to 53.8% and clarithromycin resistance in Haemophilus spp. from 3.7 to 37.5%. Resistance but also isolation rates were strongly related to azithromycin use. Resistance of 3217 S. aureus control isolates remained stable and resistance of 3257 Haemophilus controls increased, although at a slower rate than CF isolates. CONCLUSIONS: Over a 4 year period, azithromycin maintenance therapy in our CF population was associated with an increase in macrolide resistance in S. aureus and Haemophilus spp. PMID- 16469856 TI - Influence of imprecision on ROC curve analysis for cardiac markers. AB - BACKGROUND: There has been considerable debate regarding the impact of assay imprecision on the performance of cardiac biomarkers for diagnosis of acute coronary syndromes (ACS) and risk stratification for future adverse cardiac events. METHODS: Using existing data from 2 published clinical trials, we used a resampling method to statistically introduce 5%, 10%, and 20% imprecision to results for B-type natriuretic peptide (BNP) and cardiac troponin I (cTnI) and examined its impact on ROC curve analysis. RESULTS: Superimposition of artificial imprecision produced no significant difference in the area under the ROC curve observed for BNP for diagnosis of heart failure or for cTnI for 30-day risk stratification of patients with ACS. CONCLUSION: Assay imprecision does not appear to be a critical determinant in the interpretation of cardiac marker results for patients with heart disease. PMID- 16469852 TI - Proceedings of the SMBE Tri-National Young Investigators' Workshop 2005. Mutation rate and the cost of complexity. AB - Two recent theoretical studies of adaptation suggest that more complex organisms tend to adapt more slowly. Specifically, in Fisher's "geometric" model of a finite population where multiple traits are under optimizing selection, the average progress ensuing from a single mutation decreases as the number of traits increases--the "cost of complexity." Here, I draw on molecular and histological data to assess the extent to which on a large phylogenetic scale, this predicted decrease in the rate of adaptation per mutation is mitigated by an increase in the number of mutations per generation as complexity increases. As an index of complexity for multicellular organisms, I use the number of visibly distinct types of cell in the body. Mutation rate is the product of mutational target size and population mutation rate per unit target. Despite much scatter, genome size appears to be positively correlated with complexity (as indexed by cell-type number), which along with other considerations suggests that mutational target size tends to increase with complexity. In contrast, effective population mutation rate per unit target appears to be negatively correlated with complexity. The net result is that mutation rate probably does tend to increase with complexity, although probably not fast enough to eliminate the cost of complexity. PMID- 16469858 TI - Utility of thrombin-generation assay in the screening of factor V G1691A (Leiden) and prothrombin G20210A mutations and protein S deficiency. AB - BACKGROUND: The thrombin-generation assay has a variety of clinical uses, including diagnosis of thromboembolism-related disease, and particular profiles are associated with thrombophilic risk factors. The aim of this study was to evaluate the use of this assay in screening and identifying patients who require specific thrombophilic testing. METHODS: We used a 2-step approach to perform specific thrombophilic testing and thrombin-generation assays on 169 consecutive patients. The first step was to identify particular profiles of thrombin generation corresponding to each type of thrombophilic risk factor and to determine the pertinent variables related to thrombin generation. We then performed ROC curve analysis for each predefined variable to determine the relevant cutoffs for identification of patients in need of further testing (negative predictive value, 100%). RESULTS: Suggestive profiles were seen in factor V Leiden (n = 49) and prothrombin (n = 12) mutations and in protein S deficiency (n = 12). ROC curves showed that factor V Leiden may be excluded when the difference between lag times obtained in the absence and presence of activated protein C (APC) is >1.5 min and that prothrombin G20210A may also be excluded when the peak thrombin concentration is 63%. CONCLUSION: The thrombin generation assay represents a promising tool for screening thrombophilic risk factors, particularly in patients who are carriers of factor V Leiden or prothrombin G20210A mutations and patients with protein S deficiency. PMID- 16469857 TI - Total polyphenol intake estimated by a modified Folin-Ciocalteu assay of urine. AB - BACKGROUND: Plant polyphenols have been studied largely because of the possibility that they might underlie the protective effects afforded by fruit and vegetable intake against cancer and other chronic diseases. Measurement of polyphenol content excreted in urine as an indicator of polyphenol consumption may offer a routine screening method that could be used for these pathologies. METHODS: Thirty-six healthy volunteers each received 2 interventions, one with a polyphenol-rich food (cocoa beverage) and one with a polyphenol-free food (milk) as a control, in a randomized cross-over design with 1-week intervals. The total polyphenol content excreted in urine during the 6 h after consumption of the test meals was measured by a modified Folin-Ciocalteu assay after sample cleanup by solid-phase extraction. RESULTS: The mean (SD) concentrations of polyphenols excreted in the urine 6 h after consumption of the test meals differed significantly: 140.95 (49.27) mg catechin/g of creatinine after the polyphenol rich meal vs 90.43 (46.07) mg catechin/g of creatinine after the control meal (P <0.05). CONCLUSIONS: This method allows analysis of a large number of samples per day, which is ideal for use in epidemiologic studies and may enable estimation of polyphenol consumption and determination of their possible role in preventing of certain pathologies, such as cancer, cardiovascular and degenerative diseases. PMID- 16469859 TI - Antiprimer quenching-based real-time PCR and its application to the analysis of clinical cancer samples. AB - BACKGROUND: Nucleic acid amplification plays an increasingly important role in genetic analysis of clinical samples, medical diagnostics, and drug discovery. We present a novel quantitative PCR technology that combines the advantages of existing methods and allows versatile and flexible nucleic acid target quantification in clinical samples of widely different origin and quality. METHODS: We modified one of the 2 PCR primers by use of an oligonucleotide "tail" fluorescently labeled at the 5' end. An oligonucleotide complementary to this tail, carrying a 3' quenching molecule (antiprimer), was included in the reaction along with 2 primers. After primer extension, the reaction temperature was lowered such that the antiprimer hybridizes and quenches the fluorescence of the free primer but not the fluorescence of the double-stranded PCR product. The latter provides real-time fluorescent product quantification. This antiprimer based quantitative real-time PCR method (aQRT-PCR) was used to amplify and quantify minute amounts of input DNA for genes important to cancer. RESULTS: Simplex and multiplex aQRT-PCR demonstrated linear correlation (r(2) >0.995) down to a DNA input equivalent to 20 cells. Multiplex aQRT-PCR reliably identified the HER-2 gene in microdissected breast cancer samples; in formalin-fixed, paraffin embedded specimens; and in plasma circulating DNA from cancer patients. Adaptation to multiplex single-nucleotide polymorphism detection via allele specific aQRT-PCR allowed correct identification of apolipoprotein B polymorphisms in 51 of 51 human specimens. CONCLUSION: The simplicity, versatility, reliability, and low cost of aQRT-PCR make it suitable for genetic analysis of clinical specimens. PMID- 16469860 TI - Relationship between cortisol increment and basal cortisol: implications for the low-dose short adrenocorticotropic hormone stimulation test. AB - BACKGROUND: We analyzed the low-dose (1 microg) rapid adrenocorticotropic hormone test (LDST) in 17 patients with a normal hypothalamic-pituitary-adrenal axis to determine reference intervals for the LDST on the basis of poststimulation cortisol increments. METHODS: We analyzed test results for 17 patients (14 females and 3 males; age range, 18-46 years) who had received a 2-mL aliquot of low-dose (1 microg) adrenocorticotropic hormone prepared from one 250-microg vial of Synacthen diluted in 500 mL of sterile normal saline solution. Sampling took place at 0, 20, 30, and 60 min post stimulation. The cortisol increment was plotted against basal cortisol. RESULTS: We observed a marked interdependence of the basal cortisol concentration with the increase in cortisol concentration. The relationship was inverse and linear with the best fit observed at 30 min post stimulation. The lower 95% prediction limit for basal cortisol at the zero increment was 400 nmol/L with a mean concentration of 600 nmol/L. CONCLUSIONS: We propose that a peak cortisol concentration <400 nmol/L is a sufficient single criterion for abnormal adrenal function as assessed by the LDST. Concentrations of 400-600 nmol/L are in the gray area, and those >600 nmol/L confirm normal adrenal function. Repeat analyses with larger sample sizes are warranted to confirm these observations. PMID- 16469862 TI - Acid in the airways. Focus on "Hyperacidity of secreted fluid from submucosal glands in early cystic fibrosis". PMID- 16469863 TI - Role of AKT in cyclic strain-induced endothelial cell proliferation and survival. AB - Endothelial cells (ECs) are exposed to repetitive cyclic strain (CS) in vivo by the beating heart. The aim of this study was to assess the influence of CS amplitude and/or frequency on EC proliferation and survival and to determine the role of AKT in CS-induced EC proliferation and survival. Cultured bovine aortic ECs were exposed to 10% strain at a frequency of 60 (60 cpm-10%) or 100 (100 cpm 10%) cycles/min or 15.6% strain at a frequency of 60 cycles/min (60 cpm-15.6%). AKT, glycogen synthase kinase (GSK)-3beta, BAD, and cleaved caspase-3 were activated by CS in ECs. Increasing the magnitude or frequency of strain resulted in an earlier phosphorylation of GSK-3beta, although the magnitude of phosphorylation was similar. After CS at 60 cpm-10% for 24 h, the number of nontransfected ECs was significantly increased by 8.5% (P < 0.05). We found that the number of apoptotic ECs was slightly decreased with exposure to CS. ECs transfected with kinase-dead AKT (KA179) as well as plasmids containing a point mutation in the pleckstrin homology domain of AKT (RC25) not only prevented AKT, GSK-3beta, and BAD phosphorylation but also inhibited the CS-induced increase in cell number as well as the CS-induced protection against apoptosis (both P < 0.05). The ratio of 5'-bromo-2'-deoxyuridine-positive cells was increased when ECs transfected with RC25 and KA179 as well as nontransfected ECs and ECs transfected with Lipofectamine 2000 were exposed to CS. We conclude that AKT is important in enhancing the survival of ECs exposed to CS but is not involved in EC proliferation. PMID- 16469861 TI - Physiological role of ROCKs in the cardiovascular system. AB - Rho-associated kinases (ROCKs), the immediate downstream targets of RhoA, are ubiquitously expressed serine-threonine protein kinases that are involved in diverse cellular functions, including smooth muscle contraction, actin cytoskeleton organization, cell adhesion and motility, and gene expression. Recent studies have shown that ROCKs may play a pivotal role in cardiovascular diseases such as vasospastic angina, ischemic stroke, and heart failure. Indeed, inhibition of ROCKs by statins or other selective inhibitors leads to the upregulation and activation of endothelial nitric oxide synthase (eNOS) and reduction of vascular inflammation and atherosclerosis. Thus inhibition of ROCKs may contribute to some of the cholesterol-independent beneficial effects of statin therapy. Currently, two ROCK isoforms have been identified, ROCK1 and ROCK2. Because ROCK inhibitors are nonselective with respect to ROCK1 and ROCK2 and also, in some cases, may be nonspecific with respect to other ROCK-related kinases such as myristolated alanine-rich C kinase substrate (MARCKS), protein kinase A, and protein kinase C, the precise role of ROCKs in cardiovascular disease remains unknown. However, with the recent development of ROCK1- and ROCK2 knockout mice, further dissection of ROCK signaling pathways is now possible. Herein we review what is known about the physiological role of ROCKs in the cardiovascular system and speculate about how inhibition of ROCKs could provide cardiovascular benefits. PMID- 16469864 TI - Activation of the cannabinoid type-1 receptor mediates the anticonvulsant properties of cannabinoids in the hippocampal neuronal culture models of acquired epilepsy and status epilepticus. AB - Cannabinoids have been shown to have anticonvulsant properties, but no studies have evaluated the effects of cannabinoids in the hippocampal neuronal culture models of acquired epilepsy (AE) and status epilepticus (SE). This study investigated the anticonvulsant properties of the cannabinoid receptor agonist R(+)-[2,3-dihydro-5-methyl-3-[(morpholinyl)methyl]pyrrolol[1,2,3 de]-1,4 benzoxazinyl]-(1-naphthalenyl)methanone (WIN 55,212-2) in primary hippocampal neuronal culture models of both AE and SE. WIN 55,212-2 produced dose-dependent anticonvulsant effects against both spontaneous recurrent epileptiform discharges (SRED) (EC50 = 0.85 microM) and SE (EC50 = 1.51 microM), with total suppression of seizure activity at 3 microM and of SE activity at 5 microM. The anticonvulsant properties of WIN 55,212-2 in these preparations were both stereospecific and blocked by the cannabinoid type-1 (CB1) receptor antagonist N (piperidin-1-yl-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3 carboxamidehydrochloride (SR141716A; 1 microM), showing a CB1 receptor-dependent pathway. The inhibitory effect of WIN 55,212-2 against low Mg2+-induced SE is the first observation in this model of total suppression of SE by a selective pharmacological agent. The clinically used anticonvulsants phenytoin and phenobarbital were not able to abolish low Mg2+-induced SE at concentrations up to 150 microM. The results from this study show CB1 receptor-mediated anticonvulsant effects of the cannabimimetic WIN 55,212-2 against both SRED and low Mg2+-induced SE in primary hippocampal neuronal cultures and show that these in vitro models of AE and SE may represent powerful tools to investigate the molecular mechanisms mediating the effects of cannabinoids on neuronal excitability. PMID- 16469865 TI - Novel polymeric prodrug with multivalent components for cancer therapy. AB - We designed, synthesized, and evaluated in vitro and in vivo a novel targeted anticancer polymeric prodrug containing multiple copies of tumor targeting moiety [synthetic luteinizing hormone-releasing hormone (LHRH) peptide, analog of LHRH] and anticancer drug (camptothecin). One, two, or three molecules of the targeting peptide and anticancer drug were covalently conjugated with bis(2-carboxyethyl) polyethylene glycol polymer using citric acid as a multivalent spacer. We showed that LHRH peptide was bound to extracellular receptors and localized in plasma membrane of cancer cells. The designed tumor-targeted prodrug increased the solubility of anticancer drug and offered cytoplasmic and/or nuclear delivery of drug to cancer cells expressing LHRH receptors. The multicomponent prodrug containing three copies of the targeting peptide and drug was almost 100 times more cytotoxic and substantially had enhanced antitumor activity compared with the analogous nontargeted prodrug and prodrugs containing one or two copies of active components. PMID- 16469866 TI - Pharmacological and behavioral profile of N-(4-fluorophenylmethyl)-N-(1 methylpiperidin-4-yl)-N'-(4-(2-methylpropyloxy)phenylmethyl) carbamide (2R,3R) dihydroxybutanedioate (2:1) (ACP-103), a novel 5-hydroxytryptamine(2A) receptor inverse agonist. AB - The in vitro and in vivo pharmacological properties of N-(4-fluorophenylmethyl)-N (1-methylpiperidin-4-yl)-N'-(4-(2-methylpropyloxy)phenylmethyl)carbamide (2R,3R) dihydroxybutanedioate (2:1) (ACP-103) are presented. A potent 5-hydroxytryptamine (5-HT)(2A) receptor inverse agonist ACP-103 competitively antagonized the binding of [(3)H]ketanserin to heterologously expressed human 5-HT(2A) receptors with a mean pK(i) of 9.3 in membranes and 9.70 in whole cells. ACP-103 displayed potent inverse agonist activity in the cell-based functional assay receptor selection and amplification technology (R-SAT), with a mean pIC(50) of 8.7. ACP-103 demonstrated lesser affinity (mean pK(i) of 8.80 in membranes and 8.00 in whole cells, as determined by radioligand binding) and potency as an inverse agonist (mean pIC(50) 7.1 in R-SAT) at human 5-HT(2C) receptors, and lacked affinity and functional activity at 5-HT(2B) receptors, dopamine D(2) receptors, and other human monoaminergic receptors. Behaviorally, ACP-103 attenuated head-twitch behavior (3 mg/kg p.o.), and prepulse inhibition deficits (1-10 mg/kg s.c.) induced by the 5-HT(2A) receptor agonist (+/-)-2,5-dimethoxy-4-iodoamphetamine hydrochloride in rats and reduced the hyperactivity induced in mice by the N methyl-d-aspartate receptor noncompetitive antagonist 5H-dibenzo[a,d]cyclohepten 5,10-imine (dizocilpine maleate; MK-801) (0.1 and 0.3 mg/kg s.c.; 3 mg/kg p.o.), consistent with a 5-HT(2A) receptor mechanism of action in vivo and antipsychotic like efficacy. ACP-103 demonstrated >42.6% oral bioavailability in rats. Thus, ACP-103 is a potent, efficacious, orally active 5-HT(2A) receptor inverse agonist with a behavioral pharmacological profile consistent with utility as an antipsychotic agent. PMID- 16469867 TI - Amplification of the major satellite DNA family (FA-SAT) in a cat fibrosarcoma might be related to chromosomal instability. AB - Most mammalian chromosomes have satellite DNA sequences located at or near the centromeres, organized in arrays of variable size and higher order structure. The implications of these specific repetitive DNA sequences and their organization for centromere function are still quite cloudy. In contrast to most mammalian species, the domestic cat seems to have the major satellite DNA family (FA-SAT) localized primarily at the telomeres and secondarily at the centromeres of the chromosomes. In the present work, we analyzed chromosome preparations from a fibrosarcoma, in comparison with nontumor cells (epithelial tissue) from the same individual, by in situ hybridization of the FA-SAT cat satellite DNA family. This repetitive sequence was found to be amplified in the cat tumor chromosomes analyzed. The amplification of these satellite DNA sequences in the cat chromosomes with variable number and appearance (marker chromosomes) is discussed and might be related to mitotic instability, which could explain the exhibition of complex patterns of chromosome aberrations detected in the fibrosarcoma analyzed. PMID- 16469868 TI - Bcr-Abl reduces endoplasmic reticulum releasable calcium levels by a Bcl-2 independent mechanism and inhibits calcium-dependent apoptotic signaling. AB - The Bcr-Abl oncoprotein plays a major role in the development and progression of chronic myeloid leukemia (CML). Several studies have suggested that the expression levels of Bcr-Abl are elevated at disease progression to blast crisis and that this plays a significant role in the achievement of drug resistance. We have established cell lines expressing low and high levels of Bcr-Abl to study the molecular mechanisms involved in disease progression and drug resistance. It is now known that the endoplasmic reticulum (ER) can play a major role in the regulation of apoptosis. We therefore investigated whether Bcr-Abl expression modulates ER homeostasis and interferes with ER-mediated apoptotic pathways to promote survival. Bcr-Abl-expressing cells exhibit a decreased amount of free releasable calcium in the ER as well as a weaker capacitative calcium entry response, relative to parental cells. This effect is independent of Bcl-2, which is a known modulator of ER calcium homeostasis. The reduction in ER releasable calcium results in inhibition of the ER/mitochondria-coupling process and mitochondrial calcium uptake. This study demonstrates a novel downstream consequence of Bcr-Abl signaling. The ability to negate calcium-dependent apoptotic signaling is likely to be a major prosurvival mechanism in Bcr-Abl expressing cells. PMID- 16469869 TI - ICAM-2 mediates neutrophil transmigration in vivo: evidence for stimulus specificity and a role in PECAM-1-independent transmigration. AB - ICAM-2 has been implicated in leukocyte transmigration in vitro, but there is little in vivo evidence to support this. To address this, neutrophil migration was investigated in ICAM-2-deficient mice (KO) and in wild-type (WT) mice treated with an anti-ICAM-2 blocking monoclonal antibody (mAb) (3C4). In a peritonitis model, IL-1beta-induced accumulation of neutrophils was significantly reduced in mice treated with 3C4 (51% inhibition) and in KO mice (41% inhibition). In contrast, TNF-alpha- or thioglycolate-induced responses were not suppressed in KO mice. Analysis of IL-1beta-induced leukocyte responses in cremasteric venules of KO animals by intravital microscopy indicated a defect in transmigration (44% inhibition) but not rolling or adhesion. As found before, TNF-alpha-induced leukocyte transmigration was unaltered in the KO mice. WT mice treated with the anti-ICAM-2 mAb also exhibited a selective reduction in leukocyte transmigration in response to IL-1beta while an anti-ICAM-1 mAb inhibited both leukocyte adhesion and transmigration. Interestingly, mAb 3C4 significantly suppressed IL 1beta-induced neutrophil transmigration in PE-CAM-1 KO animals in the peritonitis model but not in the cremaster muscle. The findings provide direct evidence for the involvement of ICAM-2 in neutrophil transmigration in vivo, though this role appears to be stimulus specific. Furthermore, ICAM-2 appears capable of mediating PECAM-1-independent leukocyte transmigration. PMID- 16469870 TI - Proteasome activity restricts lentiviral gene transfer into hematopoietic stem cells and is down-regulated by cytokines that enhance transduction. AB - The therapeutic potential of hematopoietic stem cell (HSC) gene therapy can be fully exploited only by reaching efficient gene transfer into HSCs without compromising their biologic properties. Although HSCs can be transduced by HIV derived lentiviral vectors (LVs) in short ex vivo culture, they display low permissivity to the vector, requiring cytokine stimulation to reach high frequency transduction. Using stringent assays of competitive xenograft repopulation, we show that early-acting cytokines synergistically enhanced human HSC gene transfer by LVs without impairing engraftment and repopulation capacity. Using S-phase suicide assays, we show that transduction enhancement by cytokines was not dependent on cell cycle progression and that LVs can transduce quiescent HSCs. Pharmacologic inhibition of the proteasome during transduction dramatically enhanced HSC gene transfer, allowing the reach of very high levels of vector integration in their progeny in vivo. Thus, LVs are effectively restricted at a postentry step by the activity of this proteolytic complex. Unexpectedly, cytokine stimulation rapidly and substantially down-regulated proteasome activity in hematopoietic progenitors, highlighting one mechanism by which cytokines may enhance permissiveness to LV gene transfer. These findings demonstrate that antiviral responses ultimately mediated by proteasomes strongly limit the efficiency of HSC transduction by LVs and establish improved conditions for HSC based gene therapy. PMID- 16469872 TI - Dasatinib (BMS-354825) targets an earlier progenitor population than imatinib in primary CML but does not eliminate the quiescent fraction. AB - Dasatinib (BMS-354825), a novel dual SRC/BCR-ABL kinase inhibitor, exhibits greater potency than imatinib mesylate (IM) and inhibits the majority of kinase mutations in IM-resistant chronic myeloid leukemia (CML). We have previously demonstrated that IM reversibly blocks proliferation but does not induce apoptosis of primitive CML cells. Here, we have attempted to overcome this resistance with dasatinib. Primitive IM-resistant CML cells showed only single copy BCR-ABL but expressed significantly higher BCR-ABL transcript levels and BCR ABL protein compared with more mature CML cells (P = .031). In addition, CrKL phosphorylation was higher in the primitive CD34(+)CD38(-) than in the total CD34(+) population (P = .002). In total CD34(+) CML cells, IM inhibited phosphorylation of CrKL at 16 but not 72 hours, consistent with enrichment of an IM-resistant primitive population. CD34(+)CD38(-) CML cells proved resistant to IM-induced inhibition of CrKL phosphorylation and apoptosis, whereas dasatinib led to significant inhibition of CrKL phosphorylation. Kinase domain mutations were not detectable in either IM or dasatinib-resistant primitive CML cells. These data confirm that dasatinib is more effective than IM within the CML stem cell compartment; however, the most primitive quiescent CML cells appear to be inherently resistant to both drugs. PMID- 16469871 TI - Efficient inhibition of HIV-1 replication in human immature monocyte-derived dendritic cells by purified anti-HIV-1 IgG without induction of maturation. AB - During mucosal HIV transmission, immature dendritic cells (DCs) present in the mucosa are among the first cellular targets of the virus. Previous studies have analyzed the inhibition of HIV-1 transfer from human mature DCs to T lymphocytes by neutralizing IgG, but so far no in vitro data regarding the capacity of antibodies to inhibit HIV-1 infection of immature DCs have been reported. Here, we found an increased HIV-inhibitory activity of monoclonal IgG and purified polyclonal IgG when immature monocyte-derived dendritic cells (iMDDCs) were used as target cells instead of autologous blood lymphocytes. We showed that FcgammaRII is involved in the mechanism for inhibiting HIV-1 infection of iMDDCs by IgG, whereas no induction of maturation was detected at concentrations of IgG that result in a 90% reduction of HIV replication. After induction of FcgammaRI expression on iMDDCs by IFN-gamma, an augmentation of the HIV-inhibitory activity of IgG, related to the expression of FcgammaRI, was observed. Taken together, our results demonstrate the participation of FcgammaRs in HIV-1 inhibition by IgG when iMDDCs are the targets. We propose that IgG is able to efficiently inhibit HIV-1 replication in iMDDCs and should be one of the components to be induced by vaccination. PMID- 16469873 TI - Uncoupling of T-cell effector functions by inhibitory killer immunoglobulin-like receptors. AB - Killer immunoglobulin-like receptors (KIRs) are a family of regulatory cell surface molecules expressed on natural killer (NK) cells and memory T-cell subsets. Their ability to prevent the formation of an activation platform and to inhibit NK cell activation is the basis of the missing self model of NK cell function. The benefits of KIR expression for T-cell biology are unclear. We studied how KIR2DL2 regulates T-cell function. Engagement of KIR2DL2 by the ligand human leukocyte antigen (HLA)-Cw3 did not affect conjugate formation between CD4(+)KIR2DL2(+) T cells and superantigen-pulsed target cells or the development of mature immune synapses with lipid rafts. KIR2DL2 and the corresponding HLA-C ligand were initially recruited to the peripheral supramolecular activation cluster (pSMAC). Consequently, KIR2DL2 engagement did not inhibit the phosphorylation of early signaling proteins and T-cell-receptor (TCR)-mediated cytotoxicity or granule exocytosis. After 15-30 minutes, KIR2DL2 moved to the central supramolecular activation cluster (cSMAC), colocalizing with CD3. TCR synapses dissociated, and phosphorylated phospholipase C (PLC)-gamma1, Vav1, and extracellular signal-regulated kinase 1/2 (ERK1/2) were reduced 90 minutes after stimulation. Gene array studies documented that the inhibition of late signaling events by KIR2DL2 affected transcriptional gene activation. We propose that KIRs on memory T cells operate to uncouple effector functions by modifying the transcriptional profile while leaving granule exocytosis unabated. PMID- 16469874 TI - A prospective study of the natural history of transient leukemia (TL) in neonates with Down syndrome (DS): Children's Oncology Group (COG) study POG-9481. AB - A unique transient leukemia (TL) has been described in newborns with Down syndrome (DS; or trisomy 21 mosaics). This leukemia has a high incidence of spontaneous remission; however, early death and subsequent development of acute megakaryoblastic leukemia (AMKL) have been reported. We prospectively evaluated 48 infants with DS and TL to determine the natural history and biologic characteristics of this disease, identify the clinical characteristics associated with early death or subsequent leukemia, and assess the incidence of subsequent leukemia. Blast cells associated with TL in DS infants exhibited FAB M(7) morphology and phenotype. Most infants (74%) had trisomy 21 (or mosaicism) as the only cytogenetic abnormality in the blast cells. Most children were able to spontaneously clear peripheral blasts (89%), normalize blood counts (74%), and maintain a complete remission (64%). Early death occurred in 17% of infants and was significantly correlated with higher white blood cell count at diagnosis (P < .001), increased bilirubin and liver enzymes (P < .005), and a failure to normalize the blood count (P = .001). Recurrence of leukemia occurred in 19% of infants at a mean of 20 months. Development of leukemia was significantly correlated with karyotypic abnormalities in addition to trisomy 21 (P = .037). Ongoing collaborative clinical studies are needed to determine the optimal role of chemotherapy for infants at risk for increased mortality or disease recurrence and to further the knowledge of the unique biologic features of this TL. PMID- 16469875 TI - SHP1 tyrosine phosphatase negatively regulates NPM-ALK tyrosine kinase signaling. AB - Anaplastic large-cell lymphoma (ALCL) is frequently associated with the 2;5 translocation and expresses the NPM-ALK fusion protein, which possesses a constitutive tyrosine kinase activity. We analyzed SHP1 tyrosine phosphatase expression and activity in 3 ALK-positive ALCL cell lines (Karpas 299, Cost, and SU-DHL1) and in lymph node biopsies (n = 40). We found an inverse correlation between the level of NPM-ALK phosphorylation and SHP1 phosphatase activity. Pull down and coimmunoprecipitation experiments demonstrated a SHP1/NPM-ALK association. Furthermore, confocal microscopy performed on ALCL cell lines and biopsy specimens showed the colocalization of the 2 proteins in cytoplasmic bodies containing Y664-phosphorylated NPM-ALK. Dephosphorylation of NPM-ALK by SHP1 demonstrated that NPM-ALK was a SHP1 substrate. Downregulation of SHP1 expression by RNAi in Karpas cells led to hyperphosphorylation of NPM-ALK, STAT3 activation, and increase in cell proliferation. Furthermore, SHP1 overexpression in 3T3 fibroblasts stably expressing NPM-ALK led to the decrease of NPM-ALK phosphorylation, lower cell proliferation, and tumor progression in nude mice. These findings show that SHP1 is a negative regulator of NPM-ALK signaling. The use of tissue microarrays revealed that 50% of ALK-positive ALCLs were positive for SHP1. Our results suggest that SHP1 could be a critical enzyme in ALCL biology and a potential therapeutic target. PMID- 16469876 TI - Molecular dissection of Meis1 reveals 2 domains required for leukemia induction and a key role for Hoxa gene activation. AB - The Hoxa9 and Meis1 genes represent important oncogenic collaborators activated in a significant proportion of human leukemias with genetic alterations in the MLL gene. In this study, we show that the transforming property of Meis1 is modulated by 3 conserved domains, namely the Pbx interaction motif (PIM), the homeodomain, and the C-terminal region recently described to possess transactivating properties. Meis1 and Pbx1 interaction domain-swapping mutants are dysfunctional separately, but restore the full oncogenic activity of Meis1 when cotransduced in primary cells engineered to overexpress Hoxa9, thus implying a modular nature for PIM in Meis1-accelerated transformation. Moreover, we show that the transactivating domain of VP16 can restore, and even enhance, the oncogenic potential of the Meis1 mutant lacking the C-terminal 49 amino acids. In contrast to Meis1, the fusion VP16-Meis1 is spontaneously oncogenic, and all leukemias harbor genetic activation of endogenous Hoxa9 and/or Hoxa7, suggesting that Hoxa gene activation represents a key event required for the oncogenic activity of VP16-Meis1. PMID- 16469877 TI - C/EBPalpha determines hematopoietic cell fate in multipotential progenitor cells by inhibiting erythroid differentiation and inducing myeloid differentiation. AB - C/EBPalpha is an essential transcription factor required for myeloid differentiation. While C/EBPalpha can act as a cell fate switch to promote granulocyte differentiation in bipotential granulocyte-macrophage progenitors (GMPs), its role in regulating cell fate decisions in more primitive progenitors is not known. We found increased numbers of erythroid progenitors and erythroid cells in C/EBPalpha(-/-) fetal liver (FL). Also, enforced expression of C/EBPalpha in hematopoietic stem cells resulted in a loss of erythroid progenitors and an increase in myeloid cells by inhibition of erythroid development and inducing myeloid differentiation. Conditional expression of C/EBPalpha in murine erythroleukemia (MEL) cells induced myeloid-specific genes, while inhibiting erythroid-specific gene expression including erythropoietin receptor (EpoR), which suggests a novel mechanism to determine hematopoietic cell fate. Thus, C/EBPalpha functions in hematopoietic cell fate decisions by the dual actions of inhibiting erythroid and inducing myeloid gene expression in multipotential progenitors. PMID- 16469879 TI - Structure of the hydrophilic domain of respiratory complex I from Thermus thermophilus. AB - Respiratory complex I plays a central role in cellular energy production in bacteria and mitochondria. Its dysfunction is implicated in many human neurodegenerative diseases, as well as in aging. The crystal structure of the hydrophilic domain (peripheral arm) of complex I from Thermus thermophilus has been solved at 3.3 angstrom resolution. This subcomplex consists of eight subunits and contains all the redox centers of the enzyme, including nine iron sulfur clusters. The primary electron acceptor, flavin-mononucleotide, is within electron transfer distance of cluster N3, leading to the main redox pathway, and of the distal cluster N1a, a possible antioxidant. The structure reveals new aspects of the mechanism and evolution of the enzyme. The terminal cluster N2 is coordinated, uniquely, by two consecutive cysteines. The novel subunit Nqo15 has a similar fold to the mitochondrial iron chaperone frataxin, and it may be involved in iron-sulfur cluster regeneration in the complex. PMID- 16469878 TI - Functional analysis of zebrafish microfibril-associated glycoprotein-1 (Magp1) in vivo reveals roles for microfibrils in vascular development and function. AB - Mutations in fibrillin-1 (FBN1) result in Marfan syndrome, demonstrating a critical requirement for microfibrils in vessel structure and function. However, the identity and function of many microfibril-associated molecules essential for vascular development and function have yet to be characterized. In our morpholino based screen for members of the secretome required for vascular development, we identified a key player in microfibril formation in zebrafish embryogenesis. Microfibril-associated glycoprotein-1 (MAGP1) is a conserved protein found in mammalian and zebrafish microfibrils. Expression of magp1 mRNA is detected in microfibril-producing cells. Analysis of a functional Magp1-mRFP fusion protein reveals localization along the midline and in the vasculature during embryogenesis. Underexpression and overexpression analyses demonstrate that specific Magp1 protein levels are critical for vascular development. Integrin function is compromised in magp1 morphant embryos, suggesting that reduced integrin-matrix interaction is the main mechanism for the vascular defects in magp1 morphants. We further show that Magp1 and fibrillin-1 interact in vivo. This study implicates MAGP1 as a key player in microfibril formation and integrity during development. The essential role for MAGP1 in vascular morphogenesis and function also supports a wide range of clinical applications, including therapeutic targets in vascular disease and cardiovascular tissue engineering. PMID- 16469880 TI - Reactive and nonreactive scattering of H2 from a metal surface is electronically adiabatic. AB - The Born-Oppenheimer approximation of uncoupled electronic and nuclear motion is a standard tool of the computational chemist. However, its validity for molecule metal surface reactions, which are important to heterogeneous catalysis, has been questioned because of the possibility of electron-hole pair excitations. We have performed experiments and calculations on the scattering of molecular hydrogen from a catalytically relevant metal surface, obtaining absolute probabilities for changes in the molecule's velocity parallel to the representative Pt(111) surface. The comparison for in-plane and out-of-plane scattering and results for dissociative chemisorption in the same system show that for hydrogen-metal systems, reaction and diffractive scattering can be accurately described using the Born-Oppenheimer approximation. PMID- 16469881 TI - Progressive disruption of cellular protein folding in models of polyglutamine diseases. AB - Numerous human diseases are associated with the chronic expression of misfolded and aggregation-prone proteins. The expansion of polyglutamine residues in unrelated proteins is associated with the early onset of neurodegenerative disease. To understand how the presence of misfolded proteins leads to cellular dysfunction, we employed Caenorhabditis elegans polyglutamine aggregation models. Here, we find that polyglutamine expansions disrupted the global balance of protein folding quality control, resulting in the loss of function of diverse metastable proteins with destabilizing temperature-sensitive mutations. In turn, these proteins, although innocuous under normal physiological conditions, enhanced the aggregation of polyglutamine proteins. Thus, weak folding mutations throughout the genome can function as modifiers of polyglutamine phenotypes and toxicity. PMID- 16469882 TI - Seizing the opportunities. PMID- 16469883 TI - Scientific misconduct. Investigations document still more problems for stem cell researchers. PMID- 16469884 TI - Women's health. Study yields murky signals on low-fat diets and disease. PMID- 16469885 TI - Molecular biology. Protein tail modification opens way for gene activity. PMID- 16469886 TI - Tectonics. An early date for raising the roof of the world. PMID- 16469887 TI - Astrophysics. Dwarf galaxies may help define dark matter. PMID- 16469889 TI - Ecology. Salvage logging research continues to generate sparks. PMID- 16469888 TI - Neuroscience. Bullied mice implicate brain's reward pathway in mood disorders. PMID- 16469890 TI - Science policy. A budget with big winners and losers. PMID- 16469892 TI - Fish biology. Fishing for common ground. PMID- 16469891 TI - Anil Kakodkar interview. Breaking up (a nuclear program) is hard to do. PMID- 16469893 TI - Evolution. Darwin's place on campus is secure--but not supreme. PMID- 16469894 TI - Evolution. Is ID on the way out? PMID- 16469900 TI - Peer review and new investigators. PMID- 16469901 TI - Keeping the DSCOVR mission alive. PMID- 16469902 TI - How to measure national stereotypes? PMID- 16469904 TI - Comment on "Zircon thermometer reveals minimum melting conditions on earliest Earth" II. AB - Watson and Harrison (Reports, 6 May 2005, p. 841) interpreted low temperatures (approximately 700 degrees C) for Hadean zircons as evidence of the existence of wet, minimum-melting conditions within 200 million years of solar system formation. However, high-temperature melts (approximately 900 degrees C) are zircon-undersaturated and crystallize zircon only after substantial temperature drop during fractional crystallization. Zircon thermometry cannot distinguish between low- and high-temperature Hadean igneous sources. PMID- 16469903 TI - Comment on "Zircon thermometer reveals minimum melting conditions on earliest Earth" I. AB - Watson and Harrison (Reports, 6 May 2005, p. 841) proposed a model for early Earth magmatism based on crystallization temperatures of Hadean zircons. However, detrital zircon populations are skewed relative to the composition of their source terrains, Archaean isotopic and geochemical mantle signatures preclude reincorporation of Hadean continental crust into the early mantle, and the effects of early impacts should be considered. PMID- 16469905 TI - Ethics. Incidental findings in brain imaging research. PMID- 16469906 TI - Cell biology. Double knockout blow for caspases. PMID- 16469908 TI - Astronomy. Is the mystery of cosmic magnetic fields solved? PMID- 16469907 TI - Sociology. Experimental macro sociology: predicting the next best seller. PMID- 16469910 TI - Chemistry. Building molecules with carbon monoxide reductive coupling. PMID- 16469909 TI - Cell signaling. A sophisticated scaffold wields a new trick. PMID- 16469911 TI - GE Prize winner. How molecular motors move. PMID- 16469916 TI - Flowers and fungi use scents to mimic each other. AB - Some flowering plants mimic the scent and appearance of mushroom fruiting bodies. Fungi may also mimic flowers. In addition, infection of plants by certain fungi can direct the plant to develop nonfunctional floral-like structures that nonetheless primarily serve the reproductive advantage of the fungus. These various mimicries may serve to attract insects that in turn spread fungal spores or plant pollen, thus facilitating sexual reproduction of the cryptic organism. PMID- 16469913 TI - Gene regulatory networks and the evolution of animal body plans. AB - Development of the animal body plan is controlled by large gene regulatory networks (GRNs), and hence evolution of body plans must depend upon change in the architecture of developmental GRNs. However, these networks are composed of diverse components that evolve at different rates and in different ways. Because of the hierarchical organization of developmental GRNs, some kinds of change affect terminal properties of the body plan such as occur in speciation, whereas others affect major aspects of body plan morphology. A notable feature of the paleontological record of animal evolution is the establishment by the Early "Cambrian of virtually all phylum-level body plans. We identify a class of GRN component, the kernels" of the network, which, because of their developmental role and their particular internal structure, are most impervious to change. Conservation of phyletic body plans may have been due to the retention since pre Cambrian time of GRN kernels, which underlie development of major body parts. PMID- 16469915 TI - The molecular basis for wine grape quality--a volatile subject. AB - Volatile organic compounds are important flavor components of finished wines. In addition to winemaking practices, which shape wine quality, cultivation of the grape berries in the vineyard each season affects the production of volatile organic compounds as well as other chemical components that ultimately contribute to our perception of flavor in finished wines. By studying how berry flavor components are determined by the interplay of vine genotypes, the environment, and cultivation practices at the molecular level, scientists will develop advanced tools and knowledge that will aid viticulturalists in consistently producing balanced, flavorful berries for wine production. PMID- 16469917 TI - Biosynthesis of plant volatiles: nature's diversity and ingenuity. AB - Plant volatiles (PVs) are lipophilic molecules with high vapor pressure that serve various ecological roles. The synthesis of PVs involves the removal of hydrophilic moieties and oxidation/hydroxylation, reduction, methylation, and acylation reactions. Some PV biosynthetic enzymes produce multiple products from a single substrate or act on multiple substrates. Genes for PV biosynthesis evolve by duplication of genes that direct other aspects of plant metabolism; these duplicated genes then diverge from each other over time. Changes in the preferred substrate or resultant product of PV enzymes may occur through minimal changes of critical residues. Convergent evolution is often responsible for the ability of distally related species to synthesize the same volatile. PMID- 16469918 TI - Volatile signaling in plant-plant interactions: "talking trees" in the genomics era. AB - Plants may "eavesdrop" on volatile organic compounds (VOCs) released by herbivore attacked neighbors to activate defenses before being attacked themselves. Transcriptome and signal cascade analyses of VOC-exposed plants suggest that plants eavesdrop to prime direct and indirect defenses and to hone competitive abilities. Advances in research on VOC biosynthesis and perception have facilitated the production of plants that are genetically "deaf" to particular VOCs or "mute" in elements of their volatile vocabulary. Such plants, together with advances in VOC analytical instrumentation, will allow researchers to determine whether fluency enhances the fitness of plants in natural communities. PMID- 16469919 TI - Plant volatile compounds: sensory cues for health and nutritional value? AB - Plants produce many volatile metabolites. A small subset of these compounds is sensed by animals and humans, and the volatile profiles are defining elements of the distinct flavors of individual foods. Flavor volatiles are derived from an array of nutrients, including amino acids, fatty acids, and carotenoids. In tomato, almost all of the important flavor-related volatiles are derived from essential nutrients. The predominance of volatiles derived from essential nutrients and health-promoting compounds suggests that these volatiles provide important information about the nutritional makeup of foods. Evidence supporting a relation between volatile perception and nutrient or health value will be reviewed. PMID- 16469920 TI - The age of the Sahara desert. AB - In the Sahara region, the age of onset of the desert condition has been uncertain until now. Here we report on the discovery of 7,000,000-year-old eolian dune deposits from the northern Chad Basin. This geological archive is the oldest known evidence for desert occurrence in the Sahara. PMID- 16469922 TI - The role of pair dispersion in turbulent flow. AB - Mixing and transport in turbulent flows-which have strong local concentration fluctuations-are essential in many natural and industrial systems including reactions in chemical mixers, combustion in engines and burners, droplet formation in warm clouds, and biological odor detection and chemotaxis. Local concentration fluctuations, in turn, are intimately tied to the problem of the separation of pairs of fluid elements. We have measured this separation rate in an intensely turbulent laboratory flow and have found, in quantitative agreement with the seminal predictions of Batchelor, that the initial separation of the pair plays an important role in the subsequent spreading of the fluid elements. These results have surprising consequences for the decay of concentration fluctuations and have applications to biological and chemical systems. PMID- 16469921 TI - Reductive cyclotrimerization of carbon monoxide to the deltate dianion by an organometallic uranium complex. AB - Despite the long history of the Fischer-Tropsch reaction, carbon monoxide has proven remarkably resistant to selective homologation under mild conditions. Here, we find that an organouranium(III) complex induces efficient reductive trimerization of carbon monoxide at room temperature and pressure. The result is a triangular, cyclic C3O(2-)3, or deltate, dianion held between two uranium(IV) units. The bonding within the C3O(2-)3 unit and its coordination to the two U centers have been analyzed by x-ray diffraction and density functional theory computational studies, which show a stabilizing C-C agostic interaction between the C3 core and one U center. Solution nuclear magnetic resonance studies reveal a rapid equilibration of the deltate unit between the U centers. PMID- 16469923 TI - Late Quaternary atmospheric CH4 isotope record suggests marine clathrates are stable. AB - One explanation for the abrupt increases in atmospheric CH4, that occurred repeatedly during the last glacial cycle involves clathrate destabalization events. Because marine clathrates have a distinct deuterium/hydrogen (D/H) isotope ratio, any such destabilization event should cause the D/H ratio of atmospheric CH4 (deltaD(CH4)) to increase. Analyses of air trapped in the ice from the second Greenland ice sheet project show stable and/or decreasing deltaD(CH4) values during the end of the Younger and Older Dryas periods and one stadial period, suggesting that marine clathrates were stable during these abrupt warming episodes. Elevated glacial deltaD(CH4) values may be the result of a lower ratio of net to gross wetland CH4 emissions and an increase in petroleum based emissions. PMID- 16469924 TI - The spatial extent of 20th-century warmth in the context of the past 1200 years. AB - Periods of widespread warmth or cold are identified by positive or negative deviations that are synchronous across a number of temperature-sensitive proxy records drawn from the Northern Hemisphere. The most significant and longest duration feature during the last 1200 years is the geographical extent of warmth in the middle to late 20th century. Positive anomalies during 890 to 1170 and negative anomalies during 1580 to 1850 are consistent with the concepts of a Medieval Warm Period and a Little Ice Age, but comparison with instrumental temperatures shows the spatial extent of recent warmth to be of greater significance than that during the medieval period. PMID- 16469925 TI - Histone H4-K16 acetylation controls chromatin structure and protein interactions. AB - Acetylation of histone H4 on lysine 16 (H4-K16Ac) is a prevalent and reversible posttranslational chromatin modification in eukaryotes. To characterize the structural and functional role of this mark, we used a native chemical ligation strategy to generate histone H4 that was homogeneously acetylated at K16. The incorporation of this modified histone into nucleosomal arrays inhibits the formation of compact 30-nanometer-like fibers and impedes the ability of chromatin to form cross-fiber interactions. H4-K16Ac also inhibits the ability of the adenosine triphosphate-utilizing chromatin assembly and remodeling enzyme ACF to mobilize a mononucleosome, indicating that this single histone modification modulates both higher order chromatin structure and functional interactions between a nonhistone protein and the chromatin fiber. PMID- 16469927 TI - Translational regulators maintain totipotency in the Caenorhabditis elegans germline. AB - The molecular mechanisms that maintain totipotency of the germline are not well understood. Here, we show that two conserved translational regulators, MEX-3 and GLD-1, are essential for maintaining totipotency in the Caenorhabditis elegans germline. In mex-3 gld-1 mutants, germ cells transdifferentiate into various somatic cell types such as muscles or neurons. Our findings implicate RNA regulation in the maintenance of totipotency, suggest that multiple mechanisms maintain totipotency at different stages of germline development, and establish a genetically tractable model for studying the development of teratomas. PMID- 16469926 TI - Caspases 3 and 7: key mediators of mitochondrial events of apoptosis. AB - The current model of apoptosis holds that upstream signals lead to activation of downstream effector caspases. We generated mice deficient in the two effectors, caspase 3 and caspase 7, which died immediately after birth with defects in cardiac development. Fibroblasts lacking both enzymes were highly resistant to both mitochondrial and death receptor-mediated apoptosis, displayed preservation of mitochondrial membrane potential, and had defective nuclear translocation of apoptosis-inducing factor (AIF). Furthermore, the early apoptotic events of Bax translocation and cytochrome c release were also delayed. We conclude that caspases 3 and 7 are critical mediators of mitochondrial events of apoptosis. PMID- 16469928 TI - Experimental study of inequality and unpredictability in an artificial cultural market. AB - Hit songs, books, and movies are many times more successful than average, suggesting that "the best" alternatives are qualitatively different from "the rest"; yet experts routinely fail to predict which products will succeed. We investigated this paradox experimentally, by creating an artificial "music market" in which 14,341 participants downloaded previously unknown songs either with or without knowledge of previous participants' choices. Increasing the strength of social influence increased both inequality and unpredictability of success. Success was also only partly determined by quality: The best songs rarely did poorly, and the worst rarely did well, but any other result was possible. PMID- 16469929 TI - The nucleosomal surface as a docking station for Kaposi's sarcoma herpesvirus LANA. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) latency-associated nuclear antigen (LANA) mediates viral genome attachment to mitotic chromosomes. We find that N terminal LANA docks onto chromosomes by binding nucleosomes through the folded region of histones H2A-H2B. The same LANA residues were required for both H2A-H2B binding and chromosome association. Further, LANA did not bind Xenopus sperm chromatin, which is deficient in H2A-H2B; chromatin binding was rescued after assembly of nucleosomes containing H2A-H2B. We also describe the 2.9-angstrom crystal structure of a nucleosome complexed with the first 23 LANA amino acids. The LANA peptide forms a hairpin that interacts exclusively with an acidic H2A H2B region that is implicated in the formation of higher order chromatin structure. Our findings present a paradigm for how nucleosomes may serve as binding platforms for viral and cellular proteins and reveal a previously unknown mechanism for KSHV latency. PMID- 16469930 TI - Neurochemical modulation of response inhibition and probabilistic learning in humans. AB - Cognitive functions dependent on the prefrontal cortex, such as the ability to suppress behavior (response inhibition) and to learn from complex feedback (probabilistic learning), play critical roles in activities of daily life. To what extent do different neurochemical systems modulate these two cognitive functions? Here, using stop-signal and probabilistic learning tasks, we show a double dissociation for the involvement of noradrenaline and serotonin in human cognition. In healthy volunteers, inhibition of central noradrenaline reuptake improved response inhibition but had no effect on probabilistic learning, whereas inhibition of central serotonin reuptake impaired probabilistic learning with no effect on response inhibition. PMID- 16469932 TI - Effects of combined inhibition of ATP-sensitive potassium channels, nitric oxide, and prostaglandins on hyperemia during moderate exercise. AB - ATP-sensitive potassium (KATP) channels have been suggested to contribute to coronary and skeletal muscle vasodilation during exercise, either alone or interacting in a parallel or redundant process with nitric oxide (NO), prostaglandins (PGs), and adenosine. We tested the hypothesis that KATP channels, alone or in combination with NO and PGs, regulate exercise hyperemia in forearm muscle. Eighteen healthy young adults performed 20 min of moderate dynamic forearm exercise, with forearm blood flow (FBF) measured via Doppler ultrasound. After steady-state FBF was achieved for 5 min (saline control), the KATP inhibitor glibenclamide (Glib) was infused into the brachial artery for 5 min (10 microg.dl(-1).min(-1)), followed by saline infusion during the final 10 min of exercise (n = 9). Exercise increased FBF from 71 +/- 11 to 239 +/- 24 ml/min, and FBF was not altered by 5 min of Glib. Systemic plasma Glib levels were above the therapeutic range, and Glib increased insulin levels by approximately 50%, whereas blood glucose was unchanged (88 +/- 2 vs. 90 +/- 2 mg/dl). In nine additional subjects, Glib was followed by combined infusion of NG-nitro-L arginine methyl ester (L-NAME) plus ketorolac (to inhibit NO and PGs, respectively). As above, Glib had no effect on FBF but addition of L-NAME + ketorolac (i.e., triple blockade) reduced FBF by approximately 15% below steady state exercise levels in seven of nine subjects. Interestingly, triple blockade in two subjects caused FBF to transiently and dramatically decrease. This was followed by an acute recovery of flow above steady-state exercise values. We conclude 1) opening of KATP channels is not obligatory for forearm exercise hyperemia, and 2) triple blockade of NO, PGs, and KATP channels does not reduce hyperemia more than the inhibition of NO and PGs in most subjects. However, some subjects are sensitive to triple blockade, but they are able to restore FBF acutely during exercise. Future studies are required to determine the nature of these compensatory mechanisms in the affected individuals. PMID- 16469933 TI - Effect of short-term sprint interval training on human skeletal muscle carbohydrate metabolism during exercise and time-trial performance. AB - Our laboratory recently showed that six sessions of sprint interval training (SIT) over 2 wk increased muscle oxidative potential and cycle endurance capacity (Burgomaster KA, Hughes SC, Heigenhauser GJF, Bradwell SN, and Gibala MJ. J Appl Physiol 98: 1895-1900, 2005). The present study tested the hypothesis that short term SIT would reduce skeletal muscle glycogenolysis and lactate accumulation during exercise and increase the capacity for pyruvate oxidation via pyruvate dehydrogenase (PDH). Eight men [peak oxygen uptake (VO2 peak)=3.8+/-0.2 l/min] performed six sessions of SIT (4-7x30-s "all-out" cycling with 4 min of recovery) over 2 wk. Before and after SIT, biopsies (vastus lateralis) were obtained at rest and after each stage of a two-stage cycling test that consisted of 10 min at approximately 60% followed by 10 min at approximately 90% of VO2 peak. Subjects also performed a 250-kJ time trial (TT) before and after SIT to assess changes in cycling performance. SIT increased muscle glycogen content by approximately 50% (main effect, P=0.04) and the maximal activity of citrate synthase (posttraining: 7.8+/-0.4 vs. pretraining: 7.0+/-0.4 mol.kg protein -1.h-1; P=0.04), but the maximal activity of 3-hydroxyacyl-CoA dehydrogenase was unchanged (posttraining: 5.1+/-0.7 vs. pretraining: 4.9+/-0.6 mol.kg protein -1.h-1; P=0.76). The active form of PDH was higher after training (main effect, P=0.04), and net muscle glycogenolysis (posttraining: 100+/-16 vs. pretraining: 139+/-11 mmol/kg dry wt; P=0.03) and lactate accumulation (posttraining: 55+/-2 vs. pretraining: 63+/-1 mmol/kg dry wt; P=0.03) during exercise were reduced. TT performance improved by 9.6% after training (posttraining: 15.5+/-0.5 vs. pretraining: 17.2+/-1.0 min; P=0.006), and a control group (n=8, VO2 peak=3.9+/-0.2 l/min) showed no change in performance when tested 2 wk apart without SIT (posttraining: 18.8+/-1.2 vs. pretraining: 18.9+/-1.2 min; P=0.74). We conclude that short-term SIT improved cycling TT performance and resulted in a closer matching of glycogenolytic flux and pyruvate oxidation during submaximal exercise. PMID- 16469931 TI - Essential role of BDNF in the mesolimbic dopamine pathway in social defeat stress. AB - Mice experiencing repeated aggression develop a long-lasting aversion to social contact, which can be normalized by chronic, but not acute, administration of antidepressant. Using viral-mediated, mesolimbic dopamine pathway-specific knockdown of brain-derived neurotrophic factor (BDNF), we showed that BDNF is required for the development of this experience-dependent social aversion. Gene profiling in the nucleus accumbens indicates that local knockdown of BDNF obliterates most of the effects of repeated aggression on gene expression within this circuit, with similar effects being produced by chronic treatment with antidepressant. These results establish an essential role for BDNF in mediating long-term neural and behavioral plasticity in response to aversive social experiences. PMID- 16469934 TI - The structural basis of airways hyperresponsiveness in asthma. AB - We hypothesized that structural airway remodeling contributes to airways hyperresponsiveness (AHR) in asthma. Small, medium, and large airways were analyzed by computed tomography in 21 asthmatic volunteers under baseline conditions (FEV1 = 64% predicted) and after maximum response to albuterol (FEV1 = 76% predicted). The difference in pulmonary function between baseline and albuterol was an estimate of AHR to the baseline smooth muscle tone (BSMT). BSMT caused an increase in residual volume (RV) that was threefold greater than the decrease in forced vital capacity (FVC) because of a simultaneous increase in total lung capacity (TLC). The decrease in FVC with BSMT was the major determinant of the baseline FEV1 (P < 0.0001). The increase in RV correlated inversely with the relaxed luminal diameter of the medium airways (P = 0.009) and directly with the wall thickness of the large airways (P = 0.001). The effect of BSMT on functional residual capacity (FRC) controlled the change in TLC relative to the change in RV. When the FRC increased with RV, TLC increased and FVC was preserved. When the relaxed large airways were critically narrowed, FRC and TLC did not increase and FVC fell. With critical large airways narrowing, the FRC was already elevated from dynamic hyperinflation before BSMT and did not increase further with BSMT. FEV1/FVC in the absence of BSMT correlated directly with large airway luminal diameter and inversely with the fall in FVC with BSMT. These findings suggest that dynamic hyperinflation caused by narrowing of large airways is a major determinant of AHR in asthma. PMID- 16469935 TI - The weight loss elicited by cobalt protoporphyrin is related to decreased activity of nitric oxide synthase in the hypothalamus. AB - Administration of cobaltic protoporphyrin IX (CoPP) into the third ventricle of the brain by intracerebroventricular injection in rodents is known to result in transient hypophagia and remarkably prolonged weight loss. The mechanism of action of CoPP in eliciting these effects is unknown. It is known that nitric oxide plays a role in food intake and that the hyperphagia that results from a wide variety of genetic, physiological, and pharmacological stimuli can be blocked by the administration of inhibitors of the enzyme nitric oxide synthase (NOS). We demonstrate that intracerebroventricular administration of compounds that alter nitrergic tone can also change food ingestion and weight gain patterns in normophagic rats. We also demonstrate that CoPP decreases NOS activity but that it paradoxically increases neuronal NOS transcript expression and increases neuronal NOS protein content on Western blotting. PMID- 16469936 TI - Acclimatory-phase specificity of gene expression during the course of heat acclimation and superimposed hypohydration in the rat hypothalamus. AB - The induction of the heat-acclimated phenotype involves reprogramming the expression of genes encoding both constitutive and inducible proteins. In this investigation, we studied the global genomic response in the hypothalamus during heat acclimation, with and without combined hypohydration stress. Rats were acclimated for 2 days (STHA) or for 30 days (LTHA) at 34 degrees C. Hypohydration (10% decrease in body weight) was attained by water deprivation. 32P-labeled RNA samples from the hypothalamus were hybridized onto cDNA Atlas array (Clontech no. 1.2) membranes. Clustering and functional analyses of the expression profile of a battery of genes representing various central regulatory functions of body homeostasis demonstrated a biphasic acclimation profile with a transient upregulation of genes encoding ion channels, transporters, and transmitter signaling upon STHA. After LTHA, most genes returned to their preacclimation expression levels. In both STHA and LTHA, genes encoding hormones and neuropeptides, linked with metabolic rate and food intake, were downregulated. This genomic profile, demonstrating an enhanced transcription of genes linked with neuronal excitability during STHA and enhanced metabolic efficiency upon LTHA, is consistent with our previously established integrative acclimation model. The response to hypohydration was characterized by an upregulation of a large number of genes primarily associated with the regulation of ion channels, cell volume, and neuronal excitability. During STHA, the response was transiently desensitized, recovering upon LTHA. We conclude that hypohydration overrides the heat acclimatory status. It is notable that STHA and hypohydration gene profiles are analogous with the physiological profile described in the response to various types of brain injury. PMID- 16469937 TI - Chronic hypoxic decreases in soluble guanylate cyclase protein and enzyme activity are age dependent in fetal and adult ovine carotid arteries. AB - The present study tests the hypothesis that chronic hypoxia enhances reactivity to nitric oxide (NO) through age-dependent increases in soluble guanylate cyclase (sGC) and protein kinase G (PKG) activity. In term fetal and adult ovine carotids, chronic hypoxia had no significant effect on mRNA levels for the beta1 subunit of sGC, but depressed sGC abundance by 16% in fetal and 50% in adult arteries, through possible depression of rates of mRNA translation (15% in fetal and 50% in adult) and/or increased protein turnover. Chronic hypoxia also depressed the catalytic activity of sGC, but only in fetal arteries (63%). Total sGC activity was reduced by chronic hypoxia in both fetal (69%) and adult (37%) carotid homogenates, but this effect was not observed in intact arteries when sGC activity was measured by timed accumulation of cGMP. In intact arteries treated with 300 microM 3-isobutyl-1-methylxanthine (IBMX), chronic hypoxia dramatically enhanced sGC activity in fetal (186%) but not adult (89%) arteries. This latter observation suggests that homogenization either removed an sGC activator, released an sGC inhibitor, or altered the phosphorylation state of the enzyme, resulting in reduced activity. In the absence of IBMX, chronic hypoxia had no significant effect on rates of cGMP accumulation. Chronic hypoxia also depressed the ability of the cGMP analog, 8-(p-chlorophenylthio)-cGMP, to promote vasorelaxation in both fetal (8%) and adult (12%) arteries. Together, these results emphasize the fact that intact and homogenized artery studies of sGC activity do not always yield equivalent results. The results further suggest that enhancement of reactivity to NO by chronic hypoxia must occur upstream of PKG and can only be possible if changes in cGMP occurred in functional compartments that afforded either temporal or chemical protection to the actions of phosphodiesterase. The range and age dependence of hypoxic effects observed also suggest that some responses to hypoxia must be compensatory and homeostatic, with reactivity to NO as the primary regulated variable. PMID- 16469938 TI - Differences in the time course of proximal and distal airway response to inhaled histamine studied by synchrotron radiation CT. AB - We studied the kinetics of proximal and distal bronchial response to histamine aerosol in healthy anesthetized and mechanically ventilated rabbits up to 60 min after histamine administration using a novel xenon-enhanced synchrotron radiation computed tomography imaging technique. Individual proximal airway constriction was assessed by measuring the luminal cross-sectional area. Distal airway obstruction was estimated by measuring the ventilated alveolar area after inhaled xenon administration. Respiratory system conductance was assessed continuously. Proximal airway cross-sectional area decreased by 57% of the baseline value by 20 min and recovered gradually but incompletely within 60 min. The ventilated alveolar area decreased immediately after histamine inhalation by 55% of baseline value and recovered rapidly thereafter. The results indicate that the airway reaction to inhaled histamine and the subsequent recovery are significantly slower in proximal than in distal bronchi in healthy rabbit. The findings suggest that physiological reaction mechanisms to inhaled histamine in the airway walls of large and small bronchi are not similar. PMID- 16469939 TI - Can RGS4 polymorphisms be viewed as credible risk factors for schizophrenia? A critical review of the evidence. AB - There has been a recent explosion in the list of putative susceptibility genes for schizophrenia (SZ). These genes have been identified on the basis of presumed pathogenesis, linkage, and genetic association studies. While several promising candidates have arisen, identification of a conclusive genetic risk factor has remained elusive. The proof would be most compelling if it stemmed from all three of these domains. In this review, we consider such evidence in relation to the regulator of G-protein signaling 4 (RGS4), a gene localized to chromosome 1q23. Disorder-specific changes in RGS4 mRNA levels have been observed in post-mortem brain samples; linkage has been reported at chromosome 1q23; and several association studies have concluded that significant associations exist. The latter are supported by a recently conducted meta-analysis. Thus, there is suggestive evidence in each of these domains implicating a role for RGS4 in SZ susceptibility. However, analogous to other promising susceptibility candidates, the nature of the genetic association, the precise polymorphism(s) conferring risk, and the functional implications of sequence variation at this gene are unclear. We review the published data and place them in the context of suggested criteria for establishing a candidate gene as a credible susceptibility factor for disorders with non-Mendelian patterns of inheritance. PMID- 16469940 TI - An industry perspective on the NIMH consensus statement on negative symptoms. AB - Negative symptoms of schizophrenia remain an area of substantial unmet clinical need. By convening a consensus conference in January 2005, the NIMH has taken a leading role in stimulating a resurgence of interest in methodological considerations related to development of new medications for treating negative symptoms. One audience for this work is clinical researchers in industry. They must take ideas, like those emerging from this consensus meeting, and determine whether they can be applied to their global trials in order to meet the needs of a broad group of customers, which include patients, clinicians, regulators, and payers. This article takes the ideas that surfaced from the NIMH consensus work and interprets them in terms of issues that industry faces for its clinical trials. Particular emphasis is given to addressing hurdles to study design and analysis that come when developing broad-spectrum or adjunctive agents that may be effective for negative symptoms. PMID- 16469941 TI - Prenatal infection as a risk factor for schizophrenia. AB - Accumulating evidence suggests that prenatal exposure to infection contributes to the etiology of schizophrenia. This line of investigation has been advanced by birth cohort studies that utilize prospectively acquired data from serologic assays for infectious and immune biomarkers. These investigations have provided further support for this hypothesis and permitted the investigation of new infectious pathogens in relation to schizophrenia risk. Prenatal infections that have been associated with schizophrenia include rubella, influenza, and toxoplasmosis. Maternal cytokines, including interleukin-8, are also significantly increased in pregnancies giving rise to schizophrenia cases. Although replication of these findings is required, this body of work may ultimately have important implications for the prevention of schizophrenia, the elaboration of pathogenic mechanisms in this disorder, and investigations of gene environment interactions. PMID- 16469942 TI - Review of clinical correlates of P50 sensory gating abnormalities in patients with schizophrenia. AB - A large percentage of patients with schizophrenia are characterized by an abnormality in P50 sensory gating. This abnormality has been shown to be genetically linked to the alpha-7 nicotinic receptor and is transiently reversed by acute nicotine administration. These observations have led to the development of pharmacological treatments designed to improve sensory gating. However, if normalization of P50 gating abnormalities is to guide drug development, then it becomes important to delineate the clinical correlates of enhanced P50 gating. We conducted a review of all available articles through March 2005 that have examined this issue. We found that, despite the prominent role that P50 abnormalities have played in our understanding of schizophrenia, there is a relative dearth of data examining P50 clinical correlates. There is evidence suggestive of an association between P50 and measures of attention, and multiple studies have failed to document a cross-sectional or longitudinal relationship between P50 and positive, negative, or other symptoms. These results suggest that considerably more work needs to be done to understand and validate the clinical significance of this impairment. PMID- 16469943 TI - Health status of individuals with serious mental illness. AB - We examined indices of the health of persons with serious mental illness. A sample of 100 adults with schizophrenia and 100 with major mood disorder were recruited from randomly selected outpatients who were receiving community-based psychiatric treatment. Participants were surveyed about health indicators using items from the National Health and Nutrition Examination Study III and the National Health Interview Survey. Their responses were compared with those of matched samples from the general population surveys. A total of 1% of persons with serious mental illness, compared with 10% from the general population sample, met criteria for all 5 of selected health indicators: nonsmoker, exercise that meets recommended standards, good dentition, absence of obesity, and absence of serious medical co-occurring illness. Within the mentally ill group, educational level, but not a diagnosis of schizophrenia versus mood disorder, was independently associated with a composite measure of health behaviors. We conclude that an examination of multiple health indicators may be used to measure overall health status in persons with serious mental illness. PMID- 16469945 TI - Persistent high levels of plasma oxidized low-density lipoprotein after acute myocardial infarction predict stent restenosis. AB - OBJECTIVE: Recently, elevated levels of plasma oxidized low-density lipoprotein (LDL) have been shown to relate to plaque instability in human atherosclerotic lesions. We investigated prospectively patients admitted with acute myocardial infarction (AMI) who underwent primary coronary stenting to evaluate whether the 6-month outcome could be predicted by measuring plasma oxidized LDL (ox-LDL) levels at the time of hospital discharge. METHODS AND RESULTS: Plasma ox-LDL levels were measured in 102 patients with AMI undergoing primary coronary stenting using a highly sensitive ELISA method. Measurements were taken on admission and at discharge, and the findings related to the clinical outcome. At 6-month follow-up, angiographic stent restenosis occurred in 25 (25%) of the 102 AMI patients. Plasma ox-LDL levels at discharge were significantly (P=0.0074) higher in the restenosis group than those in the no-restenosis group (1.03+/-0.65 versus 0.61+/-0.34 ng/5 microg LDL protein). Multiple regression analysis showed that only plasma ox-LDL levels at discharge were a statistically significant independent predictor for late lumen loss after stenting (beta=0.645; P<0.0001). CONCLUSIONS: This prospective study demonstrates that persistence of an increased level of plasma ox-LDL at discharge is a strong independent predictor of stent restenosis at 6-month follow-up in AMI patients. PMID- 16469946 TI - Cardiac and vascular hypertrophy in Fabry disease: evidence for a new mechanism independent of blood pressure and glycosphingolipid deposition. AB - OBJECTIVE: Fabry disease is an X-linked disorder resulting from alpha galactosidase A deficiency. The cardiovascular findings include left ventricular hypertrophy (LVH) and increased intima-media thickness of the common carotid artery (CCA IMT). The current study examined the possible correlation between these parameters. To corroborate these clinical findings in vitro, plasma from Fabry patients was tested for possible proliferative effect on rat vascular smooth muscle cells (vascular smooth muscle cell [VSMC]) and mouse neonatal cardiomyocytes. METHODS AND RESULTS: Thirty male and 38 female patients were enrolled. LVH was found in 60% of men and 39% of women. Increased CCA IMT was equally present in males and females. There was a strong positive correlation between LV mass and CCA IMT (r2=0.27; P<0.0001). VSMC and neonatal cardiomyocyte proliferative response in vitro correlated with CCA IMT (r2=0.39; P<0.0004) and LV mass index (r2=0.19; P=0.028), respectively. CONCLUSIONS: LVH and CCA IMT occur concomitantly in Fabry suggesting common pathogenesis. The underlying cause may be a circulating growth-promoting factor whose presence has been confirmed in vitro. PMID- 16469944 TI - Can medication-free research ever be ethical in older people with psychotic disorders? PMID- 16469948 TI - Pericellular proteases in angiogenesis and vasculogenesis. AB - Pericellular proteases play an important role in angiogenesis and vasculogenesis. They comprise (membrane-type) matrix metalloproteinases [(MT-)MMPs], serine proteases, cysteine cathepsins, and membrane-bound aminopeptidases. Specific inhibitors regulate them. Major roles in initiating angiogenesis have been attributed to MT1-matrix metalloproteinase (MMP), MMP-2, and MMP-9. Whereas MT MMPs are membrane-bound by nature, MMP-2 and MMP-9 can localize to the membrane by binding to alphavbeta3-integrin and CD44, respectively. Proteases switch on neovascularization by activation, liberation, and modification of angiogenic growth factors and degradation of the endothelial and interstitial matrix. They also modify the properties of angiogenic growth factors and cytokines. Neovascularization requires cell migration, which depends on the assembly of protease-protein complexes at the migrating cell front. MT1-MMP and urokinase (u PA) form multiprotein complexes in the lamellipodia and focal adhesions of migrating cells, facilitating proteolysis and sufficient support for endothelial cell migration and survival. Excessive proteolysis causes loss of endothelial cell-matrix interaction and impairs angiogenesis. MMP-9 and cathepsin L stimulate the recruitment and action of blood- or bone-marrow-derived accessory cells that enhance angiogenesis. Proteases also generate fragments of extracellular matrix and hemostasis factors that have anti-angiogenic properties. Understanding the complexity of protease activities in angiogenesis contributes to recognizing new targets for stimulation or inhibition of neovascularization in disease. PMID- 16469947 TI - Decreased high-density lipoprotein (HDL) particle size, prebeta-, and large HDL subspecies concentration in Finnish low-HDL families: relationship with intima media thickness. AB - OBJECTIVE: High-density lipoprotein (HDL) cholesterol correlates inversely with the risk of coronary heart disease (CHD). The precise antiatherogenic mechanisms of HDL subspecies are not thoroughly elucidated. We studied the relationship between carotid intima-media thickness (IMT) and HDL subspecies distribution in Finnish families with low HDL cholesterol and premature CHD. METHODS AND RESULTS: Altogether, 148 members of Finnish low-HDL families and 133 healthy control subjects participated in our study. HDL particle size was significantly smaller in affected family members (HDL < or =10th Finnish age-sex specific percentile) compared with unaffected family members and control subjects (9.1+/-0.04 nm versus 9.5+/-0.05 nm, P<0.0001, versus 9.8+/-0.03 nm, P<0.0001 [mean+/-SE]). Large HDL2b particles as well as prebeta-HDL concentration were significantly decreased among the affected family members. Mean IMT was significantly higher in the affected family members than in the control subjects (0.85+/-0.01 mm versus 0.79+/-0.01 mm; P<0.0001). Age, HDL2b, systolic blood pressure, and prebeta-HDL were significant independent determinants of mean IMT. CONCLUSIONS: The decreased levels of HDL2b and prebeta-HDL reflect the potentially efflux-deficient HDL subspecies profile in the affected low-HDL family members. Decreased HDL particle size caused by the decrease of plasma concentration of HDL2b and decreased prebeta-HDL levels correlate with increased IMT. PMID- 16469949 TI - PKCdelta is necessary for Smad3 expression and transforming growth factor beta induced fibronectin synthesis in vascular smooth muscle cells. AB - OBJECTIVE: The purpose of these studies is to investigate the mechanism by which transforming growth factor (TGF)beta1 regulates the synthesis of the extracellular matrix protein fibronectin (FN). METHODS AND RESULTS: TGFbeta1 elicited a time-dependent induction of FN protein and mRNA in A10 rat aortic smooth muscle cells (SMCs). Ectopic expression of Smad3 in A10 cells stimulated both basal and TGFbeta1-induced FN expression, whereas expression of Smad7 eliminated the TGFbeta response. Because TGFbeta activated PKCdelta in SMCs, we tested the role of PKCdelta in regulation of FN expression. Inhibition of PKCdelta activity by rottlerin or dominant-negative adenovirus (AdPKCdelta DN) blocked TGFbeta1's induction of FN, whereas overexpression of PKCdelta enhanced TGFbeta's effect. Moreover, aortic SMCs isolated from PKCdelta(-/-) mice exhibited diminished FN induction in response to TGFbeta. Furthermore, we found that Smad3 protein and mRNA were markedly reduced in AdPKCdelta DN-treated A10 cells and in PKCdelta null cells. Finally, restoring Smad3 in rottlerin-treated A10 and PKCdelta null cells rescues the ability of TGFbeta to upregulate FN protein and mRNA expression. CONCLUSIONS: Our data suggest that TGFbeta-activated PKCdelta is critical to maintain normal expression of Smad3, which in turn is required for the induction of fibronectin. PKCdelta represents a promising target for treating the fibroproliferative response after arterial injury. PMID- 16469950 TI - High glucose activates nuclear factor of activated T cells in native vascular smooth muscle. AB - OBJECTIVE: Hyperglycemia has been suggested to play a role in the development of vascular disease associated with diabetes. Atypical Ca2+ signaling and gene expression are characteristic of vascular dysfunction; however, little is known regarding the effects of high glucose on Ca2+-dependent transcription in the vascular wall. METHODS AND RESULTS: Using confocal immunofluorescence, we show that modest elevation of extracellular glucose (ie, from 2 to 11.5 mmol/L) increased [Ca2+]i, leading to nuclear accumulation of nuclear factor of activated T cells (NFAT) in intact cerebral arteries from mouse. This was accompanied by increased NFAT-dependent transcriptional activity. Both the increase in Ca2+ and NFAT activation were prevented by the ectonucleotidase apyrase, suggesting a mechanism involving the release of extracellular nucleotides. We provide evidence that the potent vasoconstrictors and growth stimulators UTP and UDP mediate glucose-induced NFAT activation via P2Y receptors. NFAT nuclear accumulation was inhibited by the voltage-dependent Ca2+ channel blockers verapamil and nifedipine, the calcineurin inhibitor cyclosporine A, and the novel NFAT blocker A-285222. High glucose also regulated glycogen synthase kinase 3beta and c-Jun N terminal kinase activity, yielding decreased kinase activity and reduced export of NFAT from the nucleus, providing additional mechanisms underlying the glucose induced NFAT activation. CONCLUSIONS: Our results identify the calcineurin/NFAT signaling pathway as a potential metabolic sensor for the arterial smooth muscle response to high glucose. PMID- 16469951 TI - Dynamics and plasticity of Weibel-Palade bodies in endothelial cells. AB - Agonist-induced release of endothelial cell specific storage granules, designated Weibel-Palade bodies (WPBs), provides the endothelium with the ability to rapidly respond to changes in its micro-environment. Originally being defined as an intracellular storage pool for von Willebrand factor (VWF), it has recently been shown that an increasing number of other components, including P-selectin, interleukin (IL)-8, eotaxin-3, endothelin-1, and angiopoietin-2, is present within this subcellular organelle, implicating a role for WPB exocytosis in inflammation, hemostasis, regulation of vascular tone and angiogenesis. Recent studies emphasize that WPBs provide a dynamic storage compartment whose contents can be regulated depending on the presence of inflammatory mediators in the vascular micro-environment. Additionally, release of WPBs is tightly regulated and feedback mechanisms have been identified that prevent excessive release of bioactive components from this subcellular organelle. The ability to regulate both contents and exocytosis of WPBs endows these endothelial cell specific organelles with a remarkable plasticity. This is most likely needed to allow for controlled delivery of bioactive components into the circulation on vascular perturbation. PMID- 16469953 TI - Structure-related initiation of reentry by rapid pacing in monolayers of cardiac cells. AB - This study examines how a zigzag pattern of conduction, a form of structural heterogeneity frequently found in old or diseased hearts, affects the vulnerability to reentry during rapid pacing. A central rectangular island (8x4 mm) containing a predefined zigzag pattern was created in cultured isotropic monolayers of neonatal rat ventricular myocytes. Impulse propagation was optically mapped from 253 sites using voltage-sensitive dye and was anisotropic within the zigzag island. With increasing interval between neighboring transverse connections (a), relative to the distance between longitudinal strands (b), transverse conduction velocity (CV) decreased to 66+/-6%, 20+/-2%, and 15+/-2% of CV in the surrounding isotropic region, whereas longitudinal CV increased to 102+/-8%, 113+/-12%, and 131+/-23% for a:b ratios of 1:1, 1:5, and 1:9, respectively. During rapid pacing, propagation distal to the island was steered from the side of the island with more transverse connections ("dominant" side) toward the side with fewer connections ("weak" side). Increased asymmetry in the pattern accentuated this effect, and resulted in increased rate-dependent differences in CV on the 2 sides. Consequently, a functional obstacle formed on the weak side, followed by development of single loop reentry. The reentrant wave revolved around a line of block defined by the border of the island. Reentry chirality was determined by the weak side location, and the pacing rate needed to initiate reentry decreased with increased asymmetry in the pattern. In conclusion, reentry is readily induced by rapid pacing in confluent cardiac cell monolayers containing a central and asymmetric island of zigzag conduction. PMID- 16469954 TI - Soluble adenylyl cyclase reveals the significance of cAMP compartmentation on pulmonary microvascular endothelial cell barrier. AB - Subtle elevations in cAMP localized to the plasma membrane intensely strengthen endothelial barrier function. Paradoxically, pathogenic bacteria insert adenylyl cyclases (ACs) into eukaryotic cells generating a time-dependent cytosolic cAMP increase that disrupts rather than strengthens the endothelial barrier. These findings bring into question whether membrane versus cytosolic AC activity dominates in control of cell adhesion. To address this problem, a mammalian forskolin-sensitive soluble AC (sACI/II) was expressed in pulmonary microvascular endothelial cells. Forskolin stimulated this sACI/II construct generating a small cytosolic cAMP-pool that was not regulated by phosphodiesterases or Galphas. Whereas forskolin simultaneously activated the sACI/II construct and endogenous transmembrane ACs, the modest sACI/II activity overwhelmed the barrier protective effects of plasma membrane activity to induce endothelial gap formation. Retargeting sACI/II to the plasma membrane retained AC activity but protected the endothelial cell barrier. These findings demonstrate for the first time that the intracellular location of cAMP synthesis critically determines its physiological outcome. PMID- 16469952 TI - Regulation of vascular endothelial growth factor expression and vascularization in the myocardium by insulin receptor and PI3K/Akt pathways in insulin resistance and ischemia. AB - OBJECTIVE: This study characterized the role of insulin receptors and resistance on vascular endothelial growth factor (VEGF) expression and myocardial vascularization in physiological conditions and after ischemia. METHODS AND RESULTS: Cardiac microvascular density was reduced by 30% in insulin-resistant Zucker fatty rats versus lean controls. This was associated with a parallel 40% inhibition of insulin-stimulated activation of both Akt and VEGF expression in the myocardium and cardiomyocytes. In contrast, the activation of Erk1/2 by insulin remained unchanged. In cultured cardiomyocytes, insulin or insulin-like growth factor (IGF)-1 increased VEGF mRNA and protein expression by 2-fold. Inhibition of PI3K/Akt, especially Akt2-mediated cascades but not the Ras/MEK/Erk pathway, using chemical inhibitors, dominant negative adenoviral constructs, or siRNA approaches suppressed VEGF mRNA expression by insulin. Ventricular tissues from muscle insulin receptor knockout (MIRKO) mice, which lack insulin receptors in the myocardium, have significant reductions in insulin but not IGF-1 signaling, VEGF expression, and vascular density before and after ischemia versus controls. CONCLUSIONS: Insulin regulates VEGF gene expression and vascularization in the myocardium specifically via insulin receptors and the activation of PI3K/Akt pathway. Selective inhibition of this pathway may lead to the decreases in VEGF expression and capillary density in the myocardium of patients with insulin resistance. PMID- 16469955 TI - Pulsatile cardiac tissue grafts using a novel three-dimensional cell sheet manipulation technique functionally integrates with the host heart, in vivo. AB - We devised a method of fabricating easily transplantable scaffoldless 3D heart tissue, made with a novel cell-sheet (CS) technology from cultured cardiomyocytes using a fibrin polymer coated dish. In the present study, we tested in vivo electrical communication which is essential for improving heart function between the host heart and the grafted CS. The epicardial surface of the ventricle of an anesthetized open-chest nude rat was ablated by applying a heated metal. Bilayered CS was obtained from neonatal rat primary culture. CS was transplanted onto the injured myocardial surface (sMI) (sMI+sheet group). The rats were allowed to recover for 1 to 4 weeks, to stabilize the grafts. Action potentials (APs) from the excised perfused heart were monitored by the fluorescence signal of di-4ANEPPS with a high speed charge-coupled device camera. The APs were observed under epicardial pacing of the host heart or the CS grafts. The pacing threshold of the current output was measured in the sMI+sheet group and in the nongrafted sMI group at the center of the sMI and in the normal zone (Nz). Bidirectional AP propagation between the sMI and Nz was observed in the sMI+sheet group (n=14), but was blocked at the marginal area of the sMI in the sMI group (n=9). The ratio of the pacing threshold (sMI/Nz) was significantly lower in the sMI+sheet than in the sMI group (3.0+/-0.7, 19.0+/-6.1 respectively P<0.05). There were neither spontaneous nor pacing-induced arrhythmias in these two groups. Bidirectional smooth AP propagation between the host heart and the grafted CS was observed. This finding suggested functional integration of this CS graft with the host heart without serious arrhythmia. PMID- 16469956 TI - Looking beyond the lumen to predict cerebrovascular events: "the road less travelled by". PMID- 16469958 TI - Metabolic syndrome and the risk of stroke in middle-aged men. AB - BACKGROUND AND PURPOSE: The metabolic syndrome, a clustering of disturbed glucose and insulin metabolism, obesity and abdominal fat distribution, dyslipidemia, and hypertension is associated with cardiovascular diseases. The aim of this study was to examine the relationship of metabolic syndrome, as defined by National Cholesterol Education Program (NCEP) and World Health Organization (WHO) criteria, with the risk for stroke. METHODS: Population-based cohort study with an average follow-up of 14.3 years from eastern Finland. A total of 1131 men with no history of cardiovascular disease and diabetes at baseline participated. Sixty five strokes occurred, of which 47 were ischemic strokes. RESULTS: Men with the metabolic syndrome as defined by the NCEP criteria had a 2.05-fold (95% CI, 1.03 to 4.11; P=0.042) risk for all strokes and 2.41-fold (95% CI, 1.12 to 5.32; P=0.025) risk for ischemic stroke, after adjusting for socioeconomic status, smoking, alcohol, and family history of coronary heart disease. Additional adjustment for ischemic changes during exercise test, serum low-density lipoprotein cholesterol, plasma fibrinogen, energy intake for saturated fats, energy expenditure of leisure time physical activity, and white blood cell count, the results remained significant. The risk ratios among men with metabolic syndrome as defined by the WHO criteria were 1.82 (95% CI, 1.01 to 3.26; P=0.046) for all strokes and 2.16 (95% CI, 1.11 to 4.19; P=0.022) for ischemic stroke. After further adjustment, the respective risks were 2.08 (95% CI, 1.12 to 3.87; P=0.020) and 2.47 (95% CI, 1.21 to 5.07; P=0.013). CONCLUSIONS: The risk of any stroke is increased in men with metabolic syndrome, in the absence of stroke, diabetes and cardiovascular disease at baseline. Prevention of the metabolic syndrome presents a great challenge for clinicians with respect to stroke. PMID- 16469957 TI - Association between carotid plaque characteristics and subsequent ischemic cerebrovascular events: a prospective assessment with MRI--initial results. AB - BACKGROUND AND PURPOSE: MRI is able to quantify carotid plaque size and composition with good accuracy and reproducibility and provides an opportunity to prospectively examine the relationship between plaque features and subsequent cerebrovascular events. We tested the hypothesis that the characteristics of carotid plaque, as assessed by MRI, are possible predictors of future ipsilateral cerebrovascular events. METHODS: A total of 154 consecutive subjects who initially had an asymptomatic 50% to 79% carotid stenosis by ultrasound with > or =12 months of follow-up were included in this study. Multicontrast-weighted carotid MRIs were performed at baseline, and participants were followed clinically every 3 months to identify symptoms of cerebrovascular events. RESULTS: Over a mean follow-up period of 38.2 months, 12 carotid cerebrovascular events occurred ipsilateral to the index carotid artery. Cox regression analysis demonstrated a significant association between baseline MRI identification of the following plaque characteristics and subsequent symptoms during follow-up: presence of a thin or ruptured fibrous cap (hazard ratio, 17.0; P< or =0.001), intraplaque hemorrhage (hazard ratio, 5.2; P=0.005), larger mean intraplaque hemorrhage area (hazard ratio for 10 mm2 increase, 2.6; P=0.006), larger maximum %lipid-rich/necrotic core (hazard ratio for 10% increase, 1.6; P=0.004), and larger maximum wall thickness (hazard ratio for a 1-mm increase, 1.6; P=0.008). CONCLUSIONS: Among patients who initially had an asymptomatic 50% to 79% carotid stenosis, arteries with thinned or ruptured fibrous caps, intraplaque hemorrhage, larger maximum %lipid-rich/necrotic cores, and larger maximum wall thickness by MRI were associated with the occurrence of subsequent cerebrovascular events. Findings from this prospective study provide a basis for larger multicenter studies to assess the risk of plaque features for subsequent ischemic events. PMID- 16469962 TI - Chronic Chlamydia pneumoniae infection and stroke in Cameroon: a case-control study. AB - BACKGROUND AND PURPOSE: To determine the relationship between chronic Chlamydia pneumoniae infection and stroke in Cameroon. METHODS: Sixty-four consecutive stroke patients 26 to 80 years of age were enrolled at 2 tertiary hospitals in Yaounde, Cameroon, between March 2000 and December 2001 and matched for age and sex to 64 controls. We measured IgG (1/64) and IgA (1/16) titers against C pneumoniae in both patients and controls using a validated microimmunofluorescence technique. RESULTS: There was no significant difference between cases and controls with respect to hypertension (P=0.2), smoking (P=0.53), alcohol intake (P=0.8), body mass index (P=0.49), waist-to-hip ratio (P=0.14), and diabetes (P=0.76). IgA antibodies were detected in 50 (78.1%) patients and 27 (42.2%) controls (odds ratio [OR] 4.29; 95% CI, 1.84 to 11.56; P=0.0002), and IgG antibodies in 41 (64.1%) patients and 35 (54.7%) controls (OR, 1.46; 95% CI, 0.68 to 3.22; P=0.29). For confirmed thrombotic stroke, the association with IgA antibodies became stronger (OR, 21.0; 95% CI, 3.38 to 868.45; P<0.0001), but there was still no association with IgG antibodies (OR, 1.86; 95% CI, 0.69 to 5.50; P=0.18). CONCLUSIONS: Our study shows a strong statistical association between (IgA, and not IgG, as a serological marker of) chronic C pneumoniae infection and stroke for the first time in a resident indigenous African population. These findings, if confirmed, may have important policy implications (in terms of antibiotic use in stroke prevention) in sub Saharan Africa. PMID- 16469961 TI - Cerebral microhemorrhages predict new disabling or fatal strokes in patients with acute ischemic stroke or transient ischemic attack. AB - BACKGROUND AND PURPOSE: Cerebral microhemorrhages (MHs) are common among patients presenting with acute ischemic stroke and may predict both subsequent ischemic and hemorrhagic strokes. METHODS: We prospectively studied patients with and without MHs presenting within 12 hours of their ischemic stroke or transient ischemic attack (TIA). A magnetic resonance (MR) scan was performed within 24 hours of symptom(s) onset. The primary outcome was disabling or fatal stroke at 18 months. RESULTS: An MR scan was done in 236 patients with acute ischemic stroke or TIA. Forty-five (19.1%) patients had an MH on a baseline MR scan. Patients with MHs were 2.8x (10.8% versus 4.0%; P=0.036) more likely to have a subsequent disabling or fatal stroke than patients without an MH. The risk of symptomatic intracerebral hemorrhage was not statistically significant among MH and non-MH patients (3.3% versus 0.8%; P=0.31). CONCLUSIONS: The presence of cerebral MH(s) in patients with acute ischemic stroke or TIA predicts recurrent disabling and fatal strokes. This risk is mainly assumed by recurrent ischemic strokes. PMID- 16469966 TI - DNA integrity tests in clinical practice: it is not a simple matter of black and white (or red and green). PMID- 16469967 TI - Penile erection during transurethral surgery. AB - Intraoperative penile erection during endoscopic surgery, although an infrequent occurrence, is a troublesome complication and a challenge to the urologist. It is difficult to perform the procedure during penile erection, because various complications may occur. The etiology is unclear, and a number of pharmacological remedies have been discussed in the literature. Herein, we describe the treatment and outcomes for 3 patients with intraoperative penile erection and provide a brief review of the associated literature. Intraoperative penile erection is a rare event during transurethral procedures, with a frequency of approximately 0.1% in our institution. To our knowledge, no generally accepted protocols for the prevention or treatment of this phenomenon have been reported in the literature. We recommend intracorporeal injection of 250 microg of phenylephrine: detumescence occurred rapidly in all patients after a single injection. The mode of administration is simple, and no complications have been reported. PMID- 16469968 TI - Language development in deaf children's interactions with deaf and hearing adults: a Dutch longitudinal study. AB - The language development of two deaf girls and four deaf boys in Sign Language of the Netherlands (SLN) and spoken Dutch was investigated longitudinally. At the start, the mean age of the children was 3;5. All data were collected in video recorded semistructured conversations between individual children and deaf and hearing adults. We investigated the lexical richness and syntactic complexity of the children's utterances in SLN and spoken Dutch, as well as language dominance and interactional participation. Richness and complexity increase over time, as well as children's participation. An important outcome is that syntactic complexity is higher in utterances with both sign and speech. SLN does not have higher outcomes on richness or complexity, but is dominant in terms of frequency of use. PMID- 16469969 TI - The relative difficulty of signed arithmetic story problems for primary level deaf and hard-of-hearing students. AB - This study determines the relative difficulty and associated strategy use of arithmetic (addition and subtraction) story problems when presented in American Sign Language to primary level (K-3) deaf and hard-of-hearing students. Results showed that deaf and hard-of-hearing students may consider and respond to arithmetic story problems differently than their hearing peers, with the critical dimension in problem difficulty being based on the operation typically used to solve the problem, not the story within the problem. The types of strategies used by the students supported the order of problem difficulty. The visual-spatial nature of the problem presentation appeared not to assist the deaf and hard-of hearing students in solving the problems. Factors that may have contributed to this pattern of problem difficulty are discussed so that educators can better align mathematics instruction to the thinking of the deaf child. PMID- 16469970 TI - Degrade to create: developmental requirements for ubiquitin-mediated proteolysis during early C. elegans embryogenesis. AB - The ubiquitin protein conjugation system tags proteins with the small polypeptide ubiquitin. Most poly-ubiquitinated proteins are recognized and degraded by the proteasome, a large multi-subunit protease. Ubiquitin-dependent protein degradation is used as a regulatory tool for many essential processes, the best studied of which is eukaryotic cell cycle progression. More recently, genetic studies in C. elegans have identified multiple roles for the ubiquitin system in early development, where ubiquitin-dependent protein degradation governs such diverse events as passage through meiosis, cytoskeletal regulation and cell fate determination. PMID- 16469972 TI - The Drosophila formin DAAM regulates the tracheal cuticle pattern through organizing the actin cytoskeleton. AB - Formins are involved in a wide range of cellular processes that require the remodeling of the actin cytoskeleton. Here, we have analyzed a novel Drosophila formin, belonging to the recently described DAAM subfamily. In contrast to previous assumptions, we show that DAAM plays no essential role in planar cell polarity signaling, but it has striking requirements in organizing apical actin cables that define the taenidial fold pattern of the tracheal cuticle. These observations provide evidence the first time that the function of the taenidial organization is to prevent the collapse of the tracheal tubes. Our results indicate that although DAAM is regulated by RhoA, it functions upstream or parallel to the non-receptor tyrosine kinases Src42A and Tec29 to organize the actin cytoskeleton and to determine the cuticle pattern of the Drosophila respiratory system. PMID- 16469971 TI - Association of tracheal placodes with leg primordia in Drosophila and implications for the origin of insect tracheal systems. AB - Adaptation to diverse habitats has prompted the development of distinct organs in different animals to better exploit their living conditions. This is the case for the respiratory organs of arthropods, ranging from tracheae in terrestrial insects to gills in aquatic crustaceans. Although Drosophila tracheal development has been studied extensively, the origin of the tracheal system has been a long standing mystery. Here, we show that tracheal placodes and leg primordia arise from a common pool of cells in Drosophila, with differences in their fate controlled by the activation state of the wingless signalling pathway. We have also been able to elucidate early events that trigger leg specification and to show that cryptic appendage primordia are associated with the tracheal placodes even in abdominal segments. The association between tracheal and appendage primordia in Drosophila is reminiscent of the association between gills and appendages in crustaceans. This similarity is strengthened by the finding that homologues of tracheal inducer genes are specifically expressed in the gills of crustaceans. We conclude that crustacean gills and insect tracheae share a number of features that raise the possibility of an evolutionary relationship between these structures. We propose an evolutionary scenario that accommodates the available data. PMID- 16469973 TI - The innate immune system: friend and foe. PMID- 16469974 TI - Fast food and sedentary lifestyle: a combination that leads to obesity. PMID- 16469975 TI - The acute and chronic toxic effects of vitamin A. AB - The acute and chronic effects of vitamin A toxicity are well documented in the literature. Emerging evidence suggests that subtoxicity without clinical signs of toxicity may be a growing concern, because intake from preformed sources of vitamin A often exceeds the recommended dietary allowances (RDA) for adults, especially in developed countries. Osteoporosis and hip fracture are associated with preformed vitamin A intakes that are only twice the current RDA. Assessing vitamin A status in persons with subtoxicity or toxicity is complicated because serum retinol concentrations are nonsensitive indicators in this range of liver vitamin A reserves. The metabolism in well-nourished persons of preformed vitamin A, provided by either liver or supplements, has been studied by several research groups. To control vitamin A deficiency, large therapeutic doses are administered in developing countries to women and children, who often are undernourished. Nevertheless, little attention has been given to the short-term kinetics (ie, after absorption but before storage) of a large dose of vitamin A or to the short and long-term effects of such a dose given to lactating women on serum and breast-milk concentrations of retinol and its metabolites. Moreover, appropriate dosing regimens have not been systematically evaluated to ascertain the quantitative improvement in vitamin A status of the women and children who receive these supplements. The known acute and chronic effects of vitamin A toxicity have been reported previously. However, further research is needed to ascertain the areas of the world in which subclinical toxicity exists and to evaluate its effects on overall health and well-being. PMID- 16469976 TI - Associations of body fat and its changes over time with quality of life and prospective mortality in hemodialysis patients. AB - BACKGROUND: In maintenance hemodialysis (MHD) patients, a larger body size is associated with better survival but a worse self-reported quality of life (QoL). It is not clear whether muscle mass or body fat confers the survival advantage. OBJECTIVE: We hypothesized that both a low baseline body fat percentage and a loss of fat over time were independently associated with higher mortality but with a better QoL score. DESIGN: In 535 adult MHD patients, body fat was measured directly with the use of near infrared interactance and QoL was measured with a Short Form 36 questionnaire. The patients were followed for < or =30 mo. RESULTS: Across four 12% increments of body fat at baseline, the reported QoL scores were progressively lower (P < 0.01). After a multivariate adjustment for demographics and surrogates of muscle mass and inflammation (ie, midarm muscle circumference, serum creatinine, and proinflammatory cytokines), 46 patients with body fat of <12% had a death hazard ratio (HR) 4 times that of 199 patients with body fat content between 24% and 36% (HR: 4.01; 95% CI: 1.61, 9.99; P = 0.003). In 411 MHD patients whose body fat was remeasured after 6 mo, a fat loss (< or =-1%) was associated with a death risk 2 times that of patients who gained fat (> or =1%) after a multivariate adjustment (HR: 2.06; 95% CI: 1.05, 4.05; P = 0.04). CONCLUSIONS: A low baseline body fat percentage and fat loss over time are independently associated with higher mortality in MHD patients even after adjustment for demographics and surrogates of muscle mass and inflammation, whereas a tendency toward a worse QoL is reported by MHD patients with a higher body fat percentage. Obesity management in dialysis patients may need reconsideration. PMID- 16469977 TI - Effect of a high-protein breakfast on the postprandial ghrelin response. AB - BACKGROUND: The most satiating macronutrient appears to be dietary protein. Few studies have investigated the effects of dietary protein on ghrelin secretion in humans. OBJECTIVE: This study was designed to investigate whether a high-protein (HP) breakfast is more satiating than a high-carbohydrate breakfast (HC) through suppression of postprandial ghrelin concentrations or through other physiologic processes. DESIGN: Fifteen healthy men were studied in a single-blind, crossover design. Blood samples and subjective measures of satiety were assessed frequently for 3 h after the consumption of 2 isocaloric breakfasts that differed in their protein and carbohydrate content (58.1% of energy from protein and 14.1% of energy from carbohydrate compared with 19.3% of energy from protein and 47.3% of energy from carbohydrate). The gastric emptying rate was indirectly assessed with the acetaminophen absorption test. RESULTS: The HP breakfast decreased postprandial ghrelin secretion more than did the HC breakfast (P < 0.01). Ghrelin concentrations were correlated with glucose-dependent insulinotropic polypeptide (r = -0.65; 95% CI: -0.85, -0.29) and glucagon concentrations (r = -0.47; 95% CI: -0.75, -0.03). Compared with the HC breakfast, the HP breakfast increased glucagon (P < 0.0001) and cholecystokinin (P < 0.01), tended to increase glucose dependent insulinotropic polypeptide (P = 0.07) and glucagon-like peptide 1 (P = 0.10), and decreased the gastric emptying rate (P < 0.0001). Appetite ratings were not significantly different between the 2 treatments, and the HP breakfast did not significantly affect ad libitum energy intake. CONCLUSIONS: The HP breakfast decreased postprandial ghrelin concentrations more strongly over time than did the HC breakfast. High associations between ghrelin and glucose dependent insulinotropic polypeptide and glucagon suggest that stimulation of these peptides may mediate the postprandial ghrelin response. The HP breakfast also reduced gastric emptying, probably through increased secretion of cholecystokinin and glucagon-like peptide 1. PMID- 16469980 TI - Effects of butter high in ruminant trans and monounsaturated fatty acids on lipoproteins, incorporation of fatty acids into lipid classes, plasma C-reactive protein, oxidative stress, hemostatic variables, and insulin in healthy young men. AB - BACKGROUND: Evidence suggests that ruminant trans fatty acids (FAs), such as vaccenic acid, do not increase the risk of ischemic heart disease (IHD). However, the effects of ruminant trans FAs on risk markers of IHD have been poorly investigated. OBJECTIVE: The objective was to investigate the effect of butter with a naturally high content of vaccenic acid and a concomitantly higher content of monounsaturated FAs on classic and novel risk markers of IHD. DESIGN: In a double-blind, randomized, 5-wk, parallel intervention study, 42 healthy young men were given 115 g fat/d from test butter that was high in vaccenic acid (3.6 g vaccenic acid/d) or a control butter with a low content of vaccenic acid. Blood and urine samples were collected before and after the intervention. RESULTS: The intake of the vaccenic acid-rich diet resulted in 6% and 9% lower total cholesterol and plasma HDL-cholesterol concentrations, respectively, than did the intake of the control diet (P = 0.05 and 0.002, respectively), whereas the ratio of total to HDL cholesterol did not differ significantly between the groups. The FA composition of lipid classes reflected the FAs' proportion of the test butter. No other differences were observed. CONCLUSIONS: Butter high in ruminant trans and monounsaturated FAs resulted in significantly lower total and HDL cholesterol than did the control butter with higher amounts of saturated FAs. It may be that the differences were due to the greater content of monounsaturated FAs and the lesser content of saturated FAs in the butter rich in ruminant trans FAs, rather than to the content of vaccenic acid per se. PMID- 16469978 TI - Effects of dietary saturated, monounsaturated, and n-3 fatty acids on blood pressure in healthy subjects. AB - BACKGROUND: The quantity and quality of fats consumed in the diet influence the risk of cardiovascular disease (CVD). Although the effect of diet on plasma lipids and lipoproteins is well documented, less information exists on the role of fats on blood pressure (BP). OBJECTIVE: The objective was to evaluate the effects of different types of dietary fat on BP in healthy subjects. DESIGN: Healthy subjects (n = 162) were randomly assigned for 3 mo to follow 1 of 2 isoenergetic diets: 1 rich in monounsaturated fatty acids (MUFA diet) and the other rich in saturated fatty acids (SFA diet). Each group was further randomly assigned to receive supplementation with fish oil (3.6 g n-3 fatty acids/d) or placebo. RESULTS: Systolic BP (SBP) and diastolic BP (DBP) decreased with the MUFA diet [-2.2% (P = 0.009) and -3.8% (P = 0.0001), respectively] but did not change with the SFA diet [-1.0% (P = 0.2084) and -1.1% (P = 0.2116)]. The MUFA diet caused a significantly lower DBP than did the SFA diet (P = 0.0475). Interestingly, the favorable effects of MUFA on DBP disappeared at a total fat intake above the median (>37% of energy). The addition of n-3 fatty acids influenced neither SBP nor DBP. CONCLUSIONS: Changing the proportions of dietary fat by decreasing SFAs and increasing MUFAs decreased diastolic BP. Interestingly, the beneficial effect on BP induced by fat quality was negated by the consumption of a high total fat intake. The addition of n-3 fatty acids to the diet had no significant effect on BP. PMID- 16469979 TI - Effect of a low-fat diet on fatty acid composition in red cells, plasma phospholipids, and cholesterol esters: investigation of a biomarker of total fat intake. AB - BACKGROUND: The utility of fatty acids (FAs) as biomarkers of total fat intake is unknown. OBJECTIVE: We compared FA changes in red cells (RCs), plasma phospholipids (PLs), and cholesterol esters (CEs) in response to a low-fat diet (LFD) and a moderate-fat diet (MFD) and assessed whether individual or combination of FAs predict LFD. DESIGN: Postmenopausal women (n = 66) were randomly assigned to receive an LFD (17% of energy from fat) or an MFD (34% of energy from fat) for 6 wk. All foods were provided. FAs in diets and blood were determined by gas-liquid chromatography. FA changes between baseline and end of study were compared across diets by using t tests. FA predictors of an LFD were selected by logistic regression. RESULTS: Many FAs in RCs, PLs, and CEs responded differently to the 2 diets. Changes from baseline with an LFD for palmitic acid (16:0) (3-11% increase), behenic (22:0) and lignoceric (24:0) acids (3-20% decrease, in RCs and PLs only), cis-monounsaturated FA (MUFA) (25-35% increase), linoleic acid (18:2n-6) (11-13% decrease), trans octadecanoic acids (trans 18:1) (7-20% decrease), and n-6 highly unsaturated FA (HUFA) (2-8% increase) were significantly different from changes with an MFD. Individually, 18:2n-6 and trans 18:1 were strong predictors of an LFD [receiver operating characteristic (ROC) curves: 0.92-0.80). A logistic regression model with trans 18:1, 18:2n-6, and vaccenic acid (18:1n-7) predicted an LFD with high specificity and sensitivity (ROC curves: 0.99). CONCLUSIONS: Saturated FA, cisMUFA, n-6 HUFA, and exogenous FAs greatly differed in their response to the LFD and MFD. Parallel responses were observed in RCs, PLs, and CEs. A model with a combination of FAs almost perfectly differentiated the consumption of 34% fat from that of 17% fat. PMID- 16469982 TI - Optimized predictions of absolute and relative amounts of body fat from weight, height, other anthropometric predictors, and age 1. AB - BACKGROUND: Body mass index (BMI) is the dominating weight-for-height index, but its validity as a body fat (BF) index has not been properly examined. OBJECTIVES: Our aims were to establish and validate optimal weight-for-height indexes for predicting absolute and relative (percentage) amounts of BF, to examine whether other commonly available anthropometric variables or age could add to the predictive power, and to explore the upper limit for percentage BF. DESIGN: One thousand one hundred twelve randomly selected subjects, and an additional 149 obese subjects, were included in the study. The subjects were randomly allocated to either a primary study group or a validation group. BF was measured with dual energy X-ray absorptiometry. The relations between weight/heightx (W/Hx) and BF (absolute or percentage) were examined for values of the exponent x that ranged from 0.0 to 3.0. The predictive power of equations that were based on optimal weight-for-height indexes was compared with equations based on weight, height, other anthropometric variables, and age. RESULTS: Absolute BF was optimally and linearly predicted by W/H1, whereas the percentage BF was optimally and nonlinearly predicted by W/H2. The percentage BF asymptotically approached 52% in women and 56% in men. The percentage BF increased only marginally from BMI (in kg/m2) values of >35 in women and >60 in men. Predictions of absolute BF were associated with smaller errors (8.5% for men and 5.7% for women) than were predictions of percentage BF (8.7% for men and 7.9% for women). The addition of other anthropometric measurements for both men and women, and the addition of age for women only, in the regression analyses moderately reduced these errors. CONCLUSION: Our data suggest that W/H may be a more optimal weight-for-height index than is BMI, particularly at high body weights. PMID- 16469981 TI - Effect of soy protein varying in isoflavone content on serum lipids in healthy young men. AB - BACKGROUND: Previous research supports a role for soy protein in reducing serum lipids; however, few studies involved healthy male subjects or focused on soy isoflavones (or did both). OBJECTIVE: The objective was to ascertain the effects of soy protein varying in isoflavone content on serum lipids in healthy young men. DESIGN: Thirty-five males (x +/- SD age: 27.9 +/- 5.7 y) consumed milk protein isolate (MPI), low-isoflavone soy protein isolate (low-iso SPI; 1.64 +/- 0.19 mg aglycone isoflavones/d), and high-isoflavone SPI (high-iso SPI; 61.7 +/- 7.4 mg aglycone isoflavones/d) for 57 d each, separated by 4-wk washout periods, in a randomized crossover design. Blood samples were collected at the beginning and end of each treatment period, and total, LDL, and HDL cholesterol; triacylglycerols; apolipoprotein (apo) B; apo A-I; and C-reactive protein (CRP) were measured in serum. Twenty-four-hour urine samples were collected for 3 consecutive days at the end of each treatment period and analyzed for isoflavones. RESULTS: Urinary isoflavones were significantly greater with consumption of the high-iso SPI than with that of the low-iso SPI or MPI. The differences between the 3 treatments with respect to individual serum lipids were not significant, but the ratios of total to HDL cholesterol, LDL to HDL cholesterol, and apo B to apo A-I were significantly lower with both SPI treatments than with MPI treatment. CONCLUSION: Soy protein, regardless of isoflavone content, modulates serum lipid ratios in a direction beneficial for cardiovascular disease risk in healthy young men. PMID- 16469983 TI - Effects of variation in protein and carbohydrate intake on body mass and composition during energy restriction: a meta-regression 1. AB - BACKGROUND: It is unclear whether low-carbohydrate, high-protein, weight-loss diets benefit body mass and composition beyond energy restriction alone. OBJECTIVE: The objective was to use meta-regression to determine the effects of variations in protein and carbohydrate intakes on body mass and composition during energy restriction. DESIGN: English-language studies with a dietary intervention of > or =4200 kJ/d (1000 kcal/d), with a duration of > or =4 wk, and conducted in subjects aged > or =19 y were considered eligible for inclusion. A self-reported intake in conjunction with a biological marker of macronutrient intake was required as a minimum level of dietary control. A total of 87 studies comprising 165 intervention groups met the inclusion criteria. RESULTS: After control for energy intake, diets consisting of < or =35-41.4% energy from carbohydrate were associated with a 1.74 kg greater loss of body mass, a 0.69 kg greater loss of fat-free mass, a 1.29% greater loss in percentage body fat, and a 2.05 kg greater loss of fat mass than were diets with a higher percentage of energy from carbohydrate. In studies that were conducted for >12 wk, these differences increased to 6.56 kg, 1.74 kg, 3.55%, and 5.57 kg, respectively. Protein intakes of >1.05 g/kg were associated with 0.60 kg additional fat-free mass retention compared with diets with protein intakes < or =1.05 g/kg. In studies conducted for >12 wk, this difference increased to 1.21 kg. No significant effects of protein intake on loss of either body mass or fat mass were observed. CONCLUSION: Low-carbohydrate, high-protein diets favorably affect body mass and composition independent of energy intake, which in part supports the proposed metabolic advantage of these diets. PMID- 16469984 TI - Whole grains, bran, and germ in relation to homocysteine and markers of glycemic control, lipids, and inflammation 1. AB - BACKGROUND: Intake of whole grains is inversely associated with risk of diabetes and ischemic heart disease in observational studies. The lower risk associated with high whole-grain intakes may be mediated through improvements in glycemic control, lipid profiles, or reduced inflammation. OBJECTIVE: The aim was to examine whether the intake of whole grains, bran, and germ is related to homocysteine, plasma markers of glycemic control (fasting insulin, hemoglobin A1c, C-peptide, and leptin), lipids (total cholesterol, triacylglycerol, HDL cholesterol, and LDL cholesterol), and inflammation (C-reactive protein, fibrinogen, and interleukin 6). DESIGN: This was a cross-sectional study of the relations of whole grains, bran, and germ intakes with homocysteine and markers of glycemic control, lipids, and inflammation in 938 healthy men and women. RESULTS: Whole-grain intake was inversely associated with homocysteine and markers of glycemic control. Compared with participants in the bottom quintile of whole-grain intake, participants in the highest quintile had 17%, 14%, 14%, and 11% lower concentrations of homocysteine (P < 0.01), insulin (P = 0.12), C peptide (P = 0.03), and leptin (P = 0.03), respectively. Inverse associations were also observed with total cholesterol (P = 0.02), HDL cholesterol (P = 0.05), and LDL cholesterol (P = 0.10). Whole-grain intake was not associated with the markers of inflammation. Whole-grain intake was most strongly inversely associated with markers of glycemic control in this population. CONCLUSION: The results suggest a lower risk of diabetes and heart disease in persons who consume diets high in whole grains. PMID- 16469985 TI - Potato and french fry consumption and risk of type 2 diabetes in women. AB - BACKGROUND: Potatoes, a high glycemic form of carbohydrate, are hypothesized to increase insulin resistance and risk of type 2 diabetes. OBJECTIVE: The objective was to examine prospectively the relation between potato consumption and the risk of type 2 diabetes. DESIGN: We conducted a prospective study of 84,555 women in the Nurses' Health Study. At baseline, the women were aged 34-59 y, had no history of chronic disease, and completed a validated food-frequency questionnaire. The participants were followed for 20 y with repeated assessment of diet. RESULTS: We documented 4496 new cases of type 2 diabetes. Potato and french fry consumption were both positively associated with risk of type 2 diabetes after adjustment for age and dietary and nondietary factors. The multivariate relative risk (RR) in a comparison between the highest and the lowest quintile of potato intake was 1.14 (95% CI: 1.02, 1.26; P for trend = 0.009). The multivariate RR in a comparison between the highest and the lowest quintile of french fry intake was 1.21 (95% CI: 1.09, 1.33; P for trend < 0.0001). The RR of type 2 diabetes was 1.18 (95% CI: 1.03, 1.35) for 1 daily serving of potatoes and 1.16 (95% CI: 1.05, 1.29) for 2 weekly servings of french fries. The RR of type 2 diabetes for substituting 1 serving potatoes/d for 1 serving whole grains/d was 1.30 (95% CI: 1.08, 1.57). The association between potato consumption and risk of type 2 diabetes was more pronounced in obese women. CONCLUSIONS: Our findings suggest a modest positive association between the consumption of potatoes and the risk of type 2 diabetes in women. This association was more pronounced when potatoes were substituted for whole grains. PMID- 16469987 TI - Human vitamin E requirements assessed with the use of apples fortified with deuterium-labeled alpha-tocopheryl acetate. AB - BACKGROUND: Little is known about factors that modulate dietary alpha-tocopherol bioavailability. OBJECTIVES: The study aimed to assess the efficacy of vitamin E fortified apples as a low-fat vitamin E delivery system, the influence of fat on vitamin E absorption, and human vitamin E requirements by using plasma alpha tocopherol kinetics at a dosage of alpha-tocopherol found in food. DESIGN: Apples fortified with deuterium-labeled alpha-tocopheryl acetate were consumed by 5 participants at a breakfast containing 0%, 6%, or 21% kcal from fat in 3 sequential trials. The trials were separated by a 2-wk washout period. Blood samples were obtained up to 72 h, and plasma was analyzed for labeled and unlabeled alpha-tocopherol. RESULTS: Compared with observations in the 0% fat trial, the maximum observed plasma d6-alpha-tocopherol concentrations (Cmax) and the areas under the curve increased 2- and 3-fold during the 6% and 21% fat trials, respectively. The mean (+/-SD) estimated percentage d6-alpha-tocopherol absorbed increased from 10 +/- 4% during the 0% fat trial to 20 +/- 3% and 33 +/- 5% during the 6% and 21% fat trials, respectively. The mean time to Cmax (9 +/- 2 h), fractional disappearance rates (0.022 +/- 0.003 pools/d), and half-lives (32 +/- 4 h) did not differ significantly between the trials. With the use of fractional disappearance rates and baseline plasma alpha-tocopherol concentrations, the estimated daily plasma alpha-tocopherol efflux was 13-14 mg. The estimated rate of alpha-tocopherol delivery to tissues was 5 mg/d. CONCLUSIONS: Given an estimated 33% absorption, the amount of dietary vitamin E needed daily to replace irreversible losses is or =70 y. They completed a self-administered questionnaire that included questions about the frequency of green tea consumption. We evaluated cognitive function by using the Mini-Mental State Examination with cutoffs of <28, <26, and <24 and calculated multivariate-adjusted odds ratios (ORs) of cognitive impairment. RESULTS: Higher consumption of green tea was associated with a lower prevalence of cognitive impairment. At the <26 cutoff, after adjustment for potential confounders, the ORs for the cognitive impairment associated with different frequencies of green tea consumption were 1.00 (reference) for < or =3 cups/wk, 0.62 (95% CI: 0.33, 1.19) for 4-6 cups/wk or 1 cup/d, and 0.46 (95% CI: 0.30, 0.72) for > or =2 cups/d (P for trend = 0.0006). Corresponding ORs were 1.00 (reference), 0.60 (95% CI: 0.35, 1.02), and 0.87 (95% CI: 0.55, 1.38) (P for trend = 0.33) for black or oolong tea and 1.00 (reference), 1.16 (95% CI: 0.78, 1.73), and 1.03 (95% CI: 0.59, 1.80) (P for trend = 0.70) for coffee. The results were essentially the same at cutoffs of <28 and <24. CONCLUSION: A higher consumption of green tea is associated with a lower prevalence of cognitive impairment in humans. PMID- 16469996 TI - Predictors of weight gain in a Mediterranean cohort: the Seguimiento Universidad de Navarra Study 1. AB - BACKGROUND: High consumption of sugar-sweetened drinks has been associated with weight gain and obesity in the United States. This trend may also be affecting populations with different eating patterns who increasingly are adopting typical US dietary patterns. OBJECTIVE: We assessed whether the consumption of sweetened drinks and other food items increased the likelihood of weight gain in a Mediterranean population. DESIGN: This was a prospective cohort analysis of 7194 men and women with a mean age of 41 y who were followed-up for a median of 28.5 mo with mailed questionnaires. Dietary exposure was assessed with a previously validated semiquantitative food-frequency questionnaire. RESULTS: During follow up, we observed that 49.5% of the participants increased their weight (x weight gain: 0.64 kg; 95% CI: 0.55, 0.73 kg). In the participants who had gained > or =3 kg in the 5 y before baseline, the adjusted odds ratio of subsequent weight gain for the fifth quintile compared with the first quintile of sugar-sweetened soft drink consumption was 1.6 (95% CI: 1.2, 2.1; P for trend = 0.02). This association was absent in the participants who had not gained weight in the 5-y period before baseline. The consumption of hamburgers, pizza, and sausages (as a proxy for fast-food consumption) was also independently associated with weight gain (adjusted odds ratio for the fifth compared with the first quintile = 1.2; 95% CI: 1.0, 1.4; P for trend = 0.05). We also found a significant, but weaker, association between weight gain and both red meat and sweetened fruit juice consumption. CONCLUSION: In a Mediterranean cohort, particularly in the participants who had already gained weight, an increased consumption of sugar sweetened soft drinks and of hamburgers, pizza, and sausages was associated with a higher risk of additional subsequent weight gain. PMID- 16469997 TI - Schistosomiasis japonica, anemia, and iron status in children, adolescents, and young adults in Leyte, Philippines 1. AB - BACKGROUND: Observational and interventional evidence supports a relation between human schistosomiasis and anemia; however, the exact causal mechanisms remain unclear. Eggs translocating across the intestinal or bladder wall may result in extracorporeal blood loss with subsequent iron deficiency. Alternatively, anemia may result from cytokine-mediated dyserythropoiesis, as seen in anemia of inflammation. OBJECTIVES: By evaluating the cross-sectional relation between the intensity of Schistosoma japonicum infection, hemoglobin concentration, and iron status in 7-30-y-old persons from S. japonicum-endemic rice-farming villages in the province of Leyte, Philippines, we assessed the relative contribution of iron deficiency and anemia of inflammation to schistosomiasis-associated anemia. DESIGN: We enrolled 627 S. japonicum-infected and 111 S. japonicum-uninfected persons. We obtained stool samples to quantify S. japonicum infection and venous blood samples for hemograms and measures of iron status and inflammation. RESULTS: Intensity of S. japonicum infection was independently associated with hemoglobin (beta = -0.24; 95% CI: -0.31, -0.17). Persons with high-intensity infection had a greater risk of iron deficiency anemia (adjusted prevalence odds ratio: 6.6; 95% CI: 2.9, 14.7), but there was no evidence of this relation in low intensity infections. In contrast, anemia without iron deficiency was prevalent across all intensities (adjusted prevalence odds ratio: 3.8; 95% CI: 1.5, 9.5). CONCLUSIONS: Storage iron deficiency is a major contributor to anemia in high intensity S. japonicum infection. A high prevalence of anemia without iron deficiency, exclusion of other mechanisms of anemia, and the evidence of low bioavailable iron suggest that anemia of inflammation contributes to S. japonicum associated anemia at all infection intensities. PMID- 16469998 TI - Vitamin K status of healthy Japanese women: age-related vitamin K requirement for gamma-carboxylation of osteocalcin. AB - BACKGROUND: Vitamin K deficiency is associated with low bone mineral density and increased risk of bone fracture. Phylloquinone (K1) and menaquinone 4 (MK-4) and 7 (MK-7) are generally observed in human plasma; however, data are limited on their circulating concentrations and their associations with bone metabolism or with gamma-carboxylation of the osteocalcin molecule. OBJECTIVES: The objectives were to measure the circulating concentrations of K1, MK-4, and MK-7 in women and to ascertain whether each form of vitamin K is significantly associated with bone metabolism. DESIGN: Plasma concentrations of K1, MK-4, MK-7, undercarboxylated osteocalcin (ucOC; measured by using the new electrochemiluminescence immunoassay), intact osteocalcin (iOC), calcium, and phosphorus; bone-derived alkaline phosphatase activity; and concentrations of urinary creatinine, N terminal telopeptide, and deoxypyridinoline were measured in healthy women (n = 396). RESULTS: On average, MK-7 and MK-4 were the highest and lowest, respectively, of the 3 vitamers in all age groups. K1 and MK-7 correlated inversely with ucOC, but associations between nutritional basal concentration of MK-4 and ucOC were not observed. Multiple regression analysis indicated that not only K1 and MK-7 concentrations but also age were independently correlated with ucOC concentration and the ratio of ucOC to iOC. The plasma K1 or MK-7 concentration required to minimize the ucOC concentration was highest in the group aged > or =70 y, and it decreased progressively for each of the younger age groups. CONCLUSIONS: The definite role of ucOC remains unclear. However, if submaximal gamma-carboxylation is related to the prevention of fracture or bone mineral loss, circulating vitamin K concentrations in elderly people should be kept higher than those in young people. PMID- 16469999 TI - High-protein diet, obesity, and the environment. PMID- 16470002 TI - Living Well to 100: Nutrition, Genetics, Inflammation. Proceedings of a conference, Boston, Massachusetts, USA, November 15-16, 2004. PMID- 16470001 TI - Do unsaturated fatty acids function as endogenous antibacterial and antiviral molecules? PMID- 16470000 TI - Assessment of biomarker selection in selenium-deficiency disease. PMID- 16470003 TI - What is healthy aging in the 21st century? AB - In the coming years, human aging will be one of the biggest challenges faced by industrialized countries. The average life expectancy is continuously increasing and we may be faced with spending more years in poor health. Because aging is a relatively modern phenomenon, we lack knowledge for a proper understanding of this process. Current biological thinking emphasizes that organisms are encoded for early survival and reproduction, humans not excluded. Aging is not programmed nor is it inevitable. Life span is the result of the interactions between genes and the environment in which we live. In the original habitat, genes encoding early survival and reproduction were optimized in an everlasting attempt to increase fitness and prevent the species from extinction. Aging is best explained as a cost of optimizing fitness because investments in body maintenance and repair cannot be maximized. The environment also determines how a gene influencing life span is expressed over a lifetime. When the conditions in which we live significantly improve, mortality decreases, evolutionary pressures for early survival and reproduction relax, and further resources can be invested in body maintenance and repair, which increases both average life expectancy and maximum life span. Increasing our understanding of the aging process and applying available interventions will help to protect and preserve healthy aging. PMID- 16470004 TI - Evidence for nutritional benefits in prolonging wellness. AB - Healthy aging involves the interaction between genes, the environment, and lifestyle factors, particularly diet and physical activity. Worldwide, the increase in life span has led to an increase in morbidity and mortality as the result of chronic, lifestyle-influenced diseases such as type 2 diabetes, cardiovascular disease, and cancer. Nutrient deficiency diseases are giving way to energy imbalances, and links between diet and chronic disease are becoming clearer. The global demographic, epidemiologic, and nutrition transitions are dramatic and point to an urgent need to focus on preventive approaches in health care. Thus, nutrition research has shifted from focusing exclusively on alleviating nutrient deficiencies to also stressing chronic disease prevention. Ongoing initiatives to optimize long-term health and promote healthy aging are based on the concept of functional fitness, ie, the ability to lead an active and healthy life. The Dietary Reference Intakes provide a framework for assessing nutrient adequacy at the population and individual levels. In addition, the Healthy Eating Index provides a single summary measure of diet quality. To effect changes in lifestyles to optimize health as we age, health care providers need to consider all the lifestyle and environmental factors contributing to suboptimal eating and lifestyle patterns. PMID- 16470005 TI - Starting down the right path: nutrition connections with chronic diseases of later life. AB - Thirty years have been added to the average life expectancy of Americans over the past century. It is a reasonable expectation that Americans will achieve an average life span of > or =100 y within this century. The most dramatic decreases in early-life and midlife mortality coincided with advances in medicine; curative medicine has played a lesser role. The aging of the population alone has already increased health care costs, and as we move toward even longer lives, these costs will likely increase even more. Therefore, establishing and safeguarding optimal health from early life must become increasingly important concerns for governments and health care providers if they are to allocate resources wisely and ensure and maintain a high quality of life in the population. A prevention oriented, life cycle approach is critical to establishing and maintaining health throughout life. This approach can delay and compress morbidity and the social toll associated with chronic disease and disability for as long as possible into old age. Good evidence exists that early nutrition affects key risk factors for chronic degenerative diseases of middle and later life, such as osteoporosis and cardiovascular disease. The influence of nutrition on health status and morbidity supports primary, secondary, and tertiary prevention of disease and intervention strategies at each point in the process. The objective of such a prevention oriented model is to enable people to live well for longer, while minimizing chronic disability. Starting down the right path with appropriate nutrition and staying on it by eating well are important components of healthy aging. PMID- 16470006 TI - Nutritional mechanisms that influence cardiovascular disease. AB - Current evidence suggests that most significant risk factors for heart disease have been identified. Although age, sex, and genetics are important unmodifiable risk factors, most new cases of acute myocardial infarctions today can be predicted by the presence and level of 9 risk (or cardioprotective) factors that can easily be assessed and, most importantly, modified. These risk factors are the same in almost every geographic region and in every racial/ethnic group worldwide and are consistent in men and women. Eight of these 9 risk factors are influenced by diet, and most act by promoting atherogenesis, which is the most important background condition for cardiovascular disease. Dietary interventions mostly affect atherogenesis by modulating, at the cellular level, proinflammatory processes that initiate and perpetuate endothelial dysfunction, plaque formation, and, eventually, plaque rupture. For example, there is now enough evidence, both epidemiologic and clinical, of the beneficial effects of n-3 fatty acids. Either as part of a normal low-fat diet or as supplements, these fatty acids are now recommended to prevent cardiovascular disease. This review will summarize the mechanisms by which diet may influence atherogenesis through the early inception, progression, and clinical emergence of atherosclerosis, with a special focus on n 3 fatty acids. PMID- 16470007 TI - Nutritional modulators of bone remodeling during aging. AB - Bone mass declines progressively with age in both men and women from the age of approximately 30 y. Increased longevity will inevitability be associated with an increase in the incidence of osteoporosis, its associated complications, and incurred health care costs. Current pharmacologic approaches focus on inhibiting bone resorption in those with osteoporosis but do little to improve bone mass. Increased understanding of the cellular events responsible for normal bone formation has led to multiple pathways that can be targeted to positively influence bone mass. Bone morphogenetic proteins (BMPs) have been shown to stimulate bone formation, and the BMP2 gene was recently linked to osteoporosis. BMP-2 therefore represents one potential molecular target to identify new agents to simulate bone formation. Research is accumulating on the positive effects of dietary sources that stimulate the BMP2 promoter and their effects on bone formation. Flavonoids and statins occur naturally in food products and have been shown to promote bone formation. It may be possible to influence peak bone mass by dietary means and to decrease the risk of osteoporosis in later life. To ease the future burden of osteoporosis, focusing on prevention will be key, and this could include dietary interventions to stimulate bone formation. PMID- 16470008 TI - Evidence for genetic variation as a factor in maintaining health. AB - For many chronic diseases, the influence of genetics is subtle and complex and does not conform to simple Mendelian patterns of inheritance as is seen with single-gene disorders. Genetic variation can influence the propensity for the initiating event, the progression to a clinical disease state, and the trajectory of disease. One example of how genetic variations may affect complex diseases is provided by the interleukin 1 family of cytokines. This cytokine family plays a key role in mediating inflammation, which is a central component of many chronic diseases, including coronary artery disease and rheumatoid arthritis. Recent research has identified many sequence variations in the regulatory DNA of the genes coding for important members of the interleukin 1 family, and these variations are associated with differential effects on the inflammatory response. These in turn alter the risk of some diseases in which inflammation plays a role and also affect physiologic responses, such as the inflammatory response to exercise. As this new genetic knowledge is developed and extended, it may be possible to make health care interventions at an earlier stage, before clinical disease is established, rather than after tissues have been permanently damaged. PMID- 16470009 TI - Influence of human genetic variation on nutritional requirements. AB - Genetic variation is known to affect food tolerances among human subpopulations and may also influence dietary requirements, giving rise to the new field of nutritional genomics and raising the possibility of individualizing nutritional intake for optimal health and disease prevention on the basis of an individual's genome. However, because gene-diet interactions are complex and poorly understood, the use of genomic knowledge to adjust population-based dietary recommendations is not without risk. Whereas current recommendations target most of the population to prevent nutritional deficiencies, inclusion of genomic criteria may indicate subpopulations that may incur differential benefit or risk from generalized recommendations and fortification policies. Current efforts to identify gene alleles that affect nutrient utilization have been enhanced by the identification of genetic variations that have expanded as a consequence of selection under extreme conditions. Identification of genetic variation that arose as a consequence of diet as a selective pressure helps to identify gene alleles that affect nutrient utilization. Understanding the molecular mechanisms underlying gene-nutrient interactions and their modification by genetic variation is expected to result in dietary recommendations and nutritional interventions that optimize individual health. PMID- 16470010 TI - Genetic interactions with diet influence the risk of cardiovascular disease. AB - Single-nucleotide polymorphisms are an integral component of the evolutionary process that over millennia have resulted from the interaction between the environment and the human genome. Relatively recent changes in diet have upset this interaction with respect to the nutritional environment, but nutritional science is beginning to better understand the interaction between genes and diet, with the resulting potential to influence cardiovascular disease risk by dietary modification. Single-nucleotide polymorphisms in several genes have been linked to differential effects in terms of lipid metabolism; however, even a simple model of benefit and risk is difficult to interpret in terms of dietary advice to carriers of the various alleles because of conflicting interactions between different genes. The n-3 family of polyunsaturated fatty acids is underrepresented in our modern diet; much of the benefit of polyunsaturated fatty acids found in studies of various polymorphisms seems to be linked to increased n 3 polyunsaturated fatty acid intake. The nascent science of nutrigenomics faces many challenges; more and better research is needed to clarify the picture, rebut scepticism, and re-invigorate the discussion concerning genetic polymorphism and its interaction with diet. PMID- 16470011 TI - Interleukin 1 and interleukin 18 as mediators of inflammation and the aging process. AB - In this review, 2 cytokines are discussed with respect to the inflammatory processes that are fundamental to aging and mortality. Both interleukin (IL)-1 and IL-18 are members of the same structural family (IL-1 family, or IL-F); there are presently 9 members of this family, but with the exception of IL-1alpha, IL 1beta, and IL-18, the others are antagonists or remain without known function. IL 1alpha is an intracellular cytokine with properties of both a cytokine and a transcription factor. IL-1beta and IL-18 are closely related; both possess a similar three-dimensional structure, and their respective precursor forms are inactive until cleaved by the intracellular cysteine protease caspase-1. Patients with mutations in the NALP3 gene, which controls the activity of caspase-1, readily secrete more IL-1beta and IL-18 and suffer from systemic inflammatory diseases. Patients with defects in this gene have high circulating concentrations of IL-6, serum amyloid A, and C-reactive protein, each of which decrease rapidly upon blockade of the IL-1 receptor, which suggests that IL-1beta contributes to the elevation of these markers of the inflammatory mechanisms of aging. Animal studies support the concept that IL-1beta and IL-18 participate in the pathogenesis of atherosclerosis. For example, overexpression of the IL-18 binding protein, a naturally occurring, specific inhibitor of IL-18, prevents the spontaneous development of atherosclerosis in apolipoprotein E-deficient mice. From human and animal studies, one may conclude that IL-1beta and IL-18 participate in fundamental inflammatory processes that increase during the aging process. PMID- 16470012 TI - Inflammation and cardiovascular disease mechanisms. AB - The traditional view of atherosclerosis as a lipid storage disease crumbles in the face of extensive and growing evidence that inflammation participates centrally in all stages of this disease, from the initial lesion to the end-stage thrombotic complications. Investigators now appreciate that narrowing arteries do not necessarily presage myocardial infarction and that simply treating narrowed blood vessels does not prolong life. Although invasive approaches such as angioplasty and coronary artery bypass will remain necessary in some cases, we now understand that at least some of the cardiovascular benefits attributable to medical treatment and lifestyle modification (diet and physical activity) may result from reductions in inflammatory processes. PMID- 16470013 TI - Obesity and the role of adipose tissue in inflammation and metabolism. AB - Recent discoveries, notably of the hormones leptin and adiponectin, have revised the notion that adipocytes are simply a storage depot for body energy. Instead, adipocytes are also endocrine organs, with multiple metabolic roles in regulating whole-body physiology. Small adipocytes in lean individuals promote metabolic homeostasis; the enlarged adipocytes of obese individuals recruit macrophages and promote inflammation and the release of a range of factors that predispose toward insulin resistance. Exercise activates the AMP-activated protein kinase (AMPK) in muscle and other tissues, a pathway that increases fat oxidation and glucose transport. Importantly, the adipocyte hormones leptin and adiponectin also activate AMPK; remarkably, the same pathway is activated by certain antidiabetic agents such as thiazolidinediones. Increasingly, our understanding of the adipocyte as an endocrine organ is leading to new insights into obesity and health. PMID- 16470014 TI - Genetics of aging: implications for drug discovery and development. AB - Aging is not a passive activity, but an actively regulated metabolic process. Specific genes have been identified that regulate aging, although aging, and consequently longevity, is only partially under genetic influence. It is also possible to increase life span by environmental modification; for example, caloric restriction can increase life span. Because human life span is long, directly studying aging in humans is impractical. Fortunately, significant insights into aging can be achieved by studying short-lived organisms, such as yeast, worms, and fruit flies. Many of the molecular pathways regulating aging in these lower organisms are conserved in mammals and overlap with pathways regulating metabolism. For example, an insulin-growth hormone signaling system has been implicated in regulating aging and longevity in both worms and mammals. Furthermore, the dysregulation of glucose homeostasis is a hallmark of aging in humans. In fact, type 2 diabetes, a disease of glucose homeostasis, can be conceptualized as a form of accelerated aging. Consistent with this, aging and diabetes are both common risk factors for a wide range of diseases. Because aging and diabetes are intimately related at a molecular level, diabetes may be able to provide the link between disease treatment (eg, diabetes) and the prevention of age-related diseases. If specific molecular pathways controlling the rate of aging can be modulated genetically, then perhaps they can be modulated pharmacologically. These insights may ultimately have an important impact on the discovery and development of drugs to both treat and prevent a wide range of diseases. PMID- 16470016 TI - Interleukin 1 genetics, inflammatory mechanisms, and nutrigenetic opportunities to modulate diseases of aging. AB - Inflammation plays a central role in many diseases of aging, and genetic differences in the inflammatory response appear to influence different disease courses among individuals. Variations in the genes for the family of interleukin 1 (IL-1) proteins are inherited together in a small set of patterns and provide an example of the role of inflammatory genetics as a modifier of diseases of aging. The IL-1 genetic variations are associated with variation in both the inflammatory response and the clinical presentation of a range of diseases, including coronary artery disease, Alzheimer disease, gastric cancer, and periodontitis. This growing understanding of the role of genetic variation in inflammation and chronic disease presents opportunities to identify healthy persons who are at increased risk of disease and to potentially modify the trajectory of disease to prolong healthy aging. Nutrition represents one of the promising approaches to modulation of the risk of diseases of aging because of the effects of certain nutrients on gene expression. One of the most practical applications of nutritional modulation of chronic disease may be nutrients that regulate the expression of key inflammatory genes. PMID- 16470017 TI - The different paths to 100. AB - Many people believe that the older a person gets, the sicker he or she becomes. The result can be quite a pessimistic view of very old age. If this were true, most if not all centenarians would have significant disability. However, approximately 90% of centenarians in a population-based study were functionally independent at the average age of 92 y. Thus, to achieve extreme old age, a much more enabling point of view emerges: the older an individual gets, the healthier he or she has been. Centenarians thus have the potential to represent a model of relative resistance to age-related diseases and slower aging. Currently, 1 in every 10 000 persons in the United States is 100 y of age or older. This prevalence is quickly changing, however, and it is likely that most industrialized nations will soon experience twice that prevalence, or one centenarian per 5000 persons. The ability to survive to extreme old age appears to be the result of a complex combination of genetics, environment, lifestyle, and luck. Understanding the genetics of the very old, and identifying the molecular drivers of longevity (or of mortality), is a potentially powerful approach to discovering and targeting the pathways mediating aging and disease susceptibility and developing preventive and therapeutic agents that will allow more of the population to age in good health. PMID- 16470015 TI - Inflammation and neurodegenerative diseases. AB - The decline in mental fitness associated with Alzheimer disease is accompanied by physical changes in the brain, including the development of characteristic plaques and neurofibrillary tangles, but the pathogenesis of those changes is not clear. Recent work suggests that the activation of microglia in response to injury, illness, aging, or other causes begins a cascade of events that can best be characterized as an inflammatory process. This cascade is mediated at first by the proinflammatory cytokine interleukin 1, which is overexpressed by the activated microglia. Through various pathways, interleukin 1 causes neuronal death, which activates more microglia, which in turn release more interleukin 1 in a self-sustaining and self-amplifying fashion. Over a period of years, this slow, smoldering inflammation in the brain destroys sufficient neurons to cause the clinical signs of Alzheimer disease. PMID- 16470018 TI - The future of living well to 100. PMID- 16470020 TI - Vernon R Young: the man, the scientist, the visionary. PMID- 16470021 TI - New functions for amino acids: effects on gene transcription and translation. AB - Amino acids act to regulate multiple processes related to gene expression, including modulation of the function of the proteins that mediate messenger RNA (mRNA) translation. By modulating the function of translation initiation and elongation factors, amino acids regulate the translation of mRNA on a global scale and also act to cause preferential changes in the translation of mRNAs encoding particular proteins or families of proteins. However, amino acids do not directly regulate the function of translation initiation and elongation factors, but instead modulate signaling through pathways traditionally considered to be solely involved in mediating the action of hormones. The best-characterized example of amino acid-induced regulation of a signal transduction pathway is one involving a protein kinase referred to as the mammalian target of rapamycin (mTOR), through which the branched-chain amino acids, particularly leucine, act to modulate the function of proteins engaged in both global mRNA translation and the selection of specific mRNAs for translation. Less understood at this point in time is evidence suggesting that amino acids also act to regulate mRNA translation through mTOR-independent mechanisms. The goal of the present review is to briefly summarize studies, primarily those performed in the laboratories of the authors, that focus on the role of the branched-chain amino acids in the regulation of mRNA translation in skeletal muscle. PMID- 16470022 TI - Arginine: beyond protein. AB - Arginine, a semiessential or conditionally essential amino acid in humans, is one of the most metabolically versatile amino acids and serves as a precursor for the synthesis of urea, nitric oxide, polyamines, proline, glutamate, creatine, and agmatine. Arginine is metabolized through a complex and highly regulated set of pathways that remain incompletely understood at both the whole-body and the cellular levels. Adding to the metabolic complexity is the fact that limited arginine availability can selectively affect the expression of specific genes, most of which are themselves involved in some aspect of arginine metabolism. This overview highlights selected aspects of arginine metabolism, including areas in which our knowledge remains fragmentary and incomplete. PMID- 16470025 TI - Vernon R Young Family Foundation: hopes for the future. PMID- 16470024 TI - What do -omics mean for the science and policy of the nutritional sciences? AB - The development of systems biology is revolutionizing the way we are studying and learning about human health. It is a way of thinking and a systematic attempt to integrate information from several fields of study (physical, biological, chemical, engineering, etc) to develop a more kinetic and real-time understanding of complex biological processes. It uses mathematical modeling tools to chart dynamic interactions between the components of a biological system, eg, genes, transcripts, proteins, metabolites, and cells, to simulate and analyze networks and pathways and the spatial and temporal relations that exist in biological systems. The term -omics represents the rigorous study of various collections of molecules, biological processes, or physiologic functions and structures as systems, represented most prominently by genomics. In the field of nutrition, had a systems approach been applied to evaluating the effect of vitamin A status on mortality rates in young children in developing countries, it might not have taken 20 y to go from the initial epidemiologic observations to global vitamin A supplementation programs. Better understanding of the functional biology of retinoids on different tissues that mediate host resistance to infection, and their synergistic interactions in biological, metabolic, and functional terms, could have provided a plausible mechanism for the observed effect on mortality. There are 3 policy take-home messages: 1) When controversies exist, invest in the science needed to sort them out. 2) Increase the amounts of money available for health research and interventions relevant to developing countries. 3) Ensure that policymakers understand the issues and why they are important and understand the science and its relevance. PMID- 16470023 TI - Network analysis of plasma and tissue amino acids and the generation of an amino index for potential diagnostic use. AB - BACKGROUND: Few studies exist on the use of metabolic profiling of amino acids to examine underlying physiologic and disease states. OBJECTIVE: We aimed to introduce a new method for studying relations among amino acids and to generate a diagnostic index, or amino index, based on amino acid concentrations. DESIGN: For network analysis, 35 Fischer-344 rats were randomly divided into 7 groups and fed diets containing 5%, 10%, 15%, 20%, 30%, 50%, or 70% protein. Amino acid concentrations in plasma and various organs were used to derive correlation coefficients that were then used to construct correlation networks. To build a diagnostic index for diabetic rats, the plasma amino acid concentrations of diabetic and normal rats were analyzed by using a novel algorithm developed to generate amino acid-based indexes. Plasma amino acid concentrations from human growth hormone transgenic rats and insulin-treated diabetic rats were used to evaluate the index obtained for diabetes. Dimethylnitrosamine-treated Sprague Dawley rats were used to generate an index for hepatic fibrosis. RESULTS: The scatter plots of plasma amino acid concentrations showed distinct patterns in different organs that were due to the different protein contents of the diets. Network analysis showed that data-driven networks for blood and tissue could be obtained. We derived a diagnostic index for the discrimination of diabetic rats with both sensitivity and specificity >97% and another surrogate index for liver hydroxyproline with a correlation of r2= 0.85. CONCLUSIONS: Correlation-based network analysis may help to uncover specific physiologic conditions or states. A novel approach using amino acid molar ratios was shown to generate indexes that can be used to separate animal disease models and monitor the progression of a disease parameter. Some of the methods described here may be applicable to the clinical setting. PMID- 16470029 TI - President Bush's proposals for healthcare reform. PMID- 16470028 TI - The tobacco industry in developing countries. PMID- 16470026 TI - Heme catalyzes tyrosine 385 nitration and inactivation of prostaglandin H2 synthase-1 by peroxynitrite. AB - The mechanism by which the inflammatory enzyme prostaglandin H(2) synthase-1 (PGHS-1) deactivates remains undefined. This study aimed to determine the stabilizing parameters of PGHS-1 and identify factors leading to deactivation by nitric oxide species (NO(x)). Purified PGHS-1 was stabilized when solubilized in beta-octylglucoside (rather than Tween-20 or CHAPS) and when reconstituted with hemin chloride (rather than hematin). Peroxynitrite (ONOO(-)) activated the peroxidase site of PGHS-1 independently of the cyclooxygenase site. After ONOO(-) exposure, holoPGHS-1 could not metabolize arachidonic acid and was structurally compromised, whereas apoPGHS-1 retained full activity once reconstituted with heme. After incubation of holoPGHS-1 with ONOO(-), heme absorbance was diminished but to a lesser extent than the loss in enzymatic function, suggesting the contribution of more than one process to enzyme inactivation. Hydroperoxide scavengers improved enzyme activity, whereas hydroxyl radical scavengers provided no protection from the effects of ONOO(-). Mass spectral analyses revealed that tyrosine 385 (Tyr 385) is a target for nitration by ONOO(-) only when heme is present. Multimer formation was also observed and required heme but could be attenuated by arachidonic acid substrate. We conclude that the heme plays a role in catalyzing Tyr 385 nitration by ONOO(-) and the demise of PGHS-1. PMID- 16470030 TI - A new direction for NHS community services. PMID- 16470027 TI - Aminophospholipid glycation and its inhibitor screening system: a new role of pyridoxal 5'-phosphate as the inhibitor. AB - Peroxidized phospholipid-mediated cytotoxity is involved in the pathophysiology of a number of diseases [i.e., the abnormal increase of phosphatidylcholine hydroperoxide (PCOOH) found in the plasma of type 2 diabetic patients]. The PCOOH accumulation may relate to Amadori-glycated phosphatidylethanolamine (deoxy-D fructosyl PE, or Amadori-PE), because Amadori-PE causes oxidative stress. However, lipid glycation inhibitor has not been discovered yet because of the lack of a lipid glycation model useful for inhibitor screening. We optimized and developed a lipid glycation model considering various reaction conditions (glucose concentration, temperature, buffer type, and pH) between PE and glucose. Using the developed model, various protein glycation inhibitors (aminoguanidine, pyridoxamine, and carnosine), antioxidants (ascorbic acid, alpha-tocopherol, quercetin, and rutin), and other food compounds (L-lysine, L-cysteine, pyridoxine, pyridoxal, and pyridoxal 5'-phosphate) were evaluated for their antiglycative properties. Pyridoxal 5'-phosphate and pyridoxal (vitamin B(6) derivatives) were the most effective antiglycative compounds. These pyridoxals could easily be condensed with PE before the glucose/PE reaction occurred. Because PE-pyridoxal 5'-phosphate adduct was detectable in human red blood cells and the increased plasma Amadori-PE concentration in streptozotocin-induced diabetic rats was decreased by dietary supplementation of pyridoxal 5'-phosphate, it is likely that pyridoxal 5'-phosphate acts as a lipid glycation inhibitor in vivo, which possibly contributes to diabetes prevention. PMID- 16470031 TI - Is amiodarone safe in heart failure? PMID- 16470032 TI - Research and development in stroke services. PMID- 16470033 TI - Half of extra money for the NHS has gone on pay rises. PMID- 16470035 TI - Patient takes her fight for trastuzumab to the High Court. PMID- 16470037 TI - Inhaled insulin is approved in Europe and United States. PMID- 16470038 TI - Doctors with history of drug misuse will face spot checks. PMID- 16470039 TI - UK report supports EU proposals to test drugs in children. PMID- 16470040 TI - Italy introduces "baby boxes" to save lives of abandoned newborns. PMID- 16470042 TI - UK defies US by funding abortion programmes. PMID- 16470048 TI - Spain's smoking restrictions spark tobacco price war. PMID- 16470045 TI - Watchdog rules that Merck violated the UK industry code of practice. PMID- 16470049 TI - Scientists refute claims that pesticides cause ill health. PMID- 16470056 TI - Identifying a hepatitis B outbreak by molecular surveillance: a case study. PMID- 16470055 TI - The first seizure and its management in adults and children. PMID- 16470057 TI - Opiate toxicity in patients with renal failure. PMID- 16470058 TI - Venous and arterial leg ulcers. PMID- 16470060 TI - Mexico and the tobacco industry: doing the wrong thing for the right reason? PMID- 16470062 TI - Applying clinical epidemiological methods to health equity: the equity effectiveness loop. PMID- 16470061 TI - British American Tobacco's erosion of health legislation in Uzbekistan. PMID- 16470063 TI - Sex workers to pay the price: a street sex worker responds to new government strategy. PMID- 16470064 TI - Sex workers to pay the price: prostitution strategy is a missed opportunity. PMID- 16470065 TI - Partial smoking ban would worsen health inequalities. PMID- 16470066 TI - Turning round NHS deficits: clinical governance, 1998-2006: RIP. PMID- 16470067 TI - Turning round NHS deficits: deficits are not for turning. PMID- 16470069 TI - Turning round NHS deficits: ...is more difficult for PCTs. PMID- 16470068 TI - Turning round NHS deficits: who says there are surplus hospital beds? PMID- 16470070 TI - Diagnostics is not Cinderella of health technology assessment. PMID- 16470071 TI - Venous thromboembolism: heparins are of porcine origin. PMID- 16470072 TI - Venous thromboembolism: stockings are important. PMID- 16470073 TI - Venous thromboembolism: potentially dangerous diagnostic pitfalls arise from diagnostic tests. PMID- 16470074 TI - Research governance is important. PMID- 16470076 TI - Diuretics, hypokalemia, and cardiac arrhythmia: a 20-year controversy. PMID- 16470075 TI - Students validate problem based learning. PMID- 16470077 TI - Prevalence, awareness, treatment, and control of hypertension: United States National Health and Nutrition Examination Survey 2001-2002. AB - The prevalence, awareness, treatment, and control of hypertension in the United States are analyzed using the National Health and Nutrition Examination Survey (NHANES) database covering the period 1988-2002. Mean body mass index was 26.1+/ 0.1 kg/m2 in 1988-1991 and 27.9+/-0.2 kg/m2 in 2001-2002 (p < 0.001). In the same period, the prevalence of diabetes mellitus increased from 5.0% to 6.5% (p = 0.03). Diastolic blood pressure was 73.3+/-0.2 mm Hg in 1988-1991 and 71.6+/-0.4 mm Hg in 2001-2002 (p < 0.001). Among the 18-39 years and 60 years and older age groups, the prevalence of hypertension increased significantly since 1988-1991. Multiple regression shows age, body mass index, and being non-Hispanic black were significantly associated with hypertension. In the period 1988-2002, the percentage receiving treatment and the percentage with blood pressure controlled increased significantly. In 2001-2002, significantly more people with hypertension and diabetes reached a blood pressure target of <130/85 mm Hg. Overall, the control rates were low, especially among middle-aged Mexican American men (8%). PMID- 16470078 TI - Providing end-organ protection with renin-angiotensin system inhibition: the evidence so far. AB - Much debate surrounds the question of the optimal therapeutic choices for medication to control blood pressure and reduce cardiovascular events. Experimental evidence suggests that drugs that block the renin-angiotensin system retard vascular disease through their direct ability to antagonize the effects of angiotensin II, which has vasoconstrictive, vascular proliferative, and atherosclerotic effects. It is not known how to separate the potential vascular protective effects of the drugs from their antihypertensive benefits. Clinical trial evidence indicates that achieved lower blood pressure goals almost always confer cardiovascular risk reduction advantages. There is also evidence, however, that successful antihypertensive regimens incorporating a renin-angiotensin system blocker, such as an angiotensin-converting enzyme inhibitor or an angiotensin receptor blocker, provide more cardiovascular risk reduction benefit compared with regimens that do not incorporate a renin-angiotensin blocker. This includes composite or specific end points involving reduction of stroke, myocardial infarction, or development of end-stage renal disease. PMID- 16470079 TI - Management strategies for patients with hypertension and diabetes: why combination therapy is critical. AB - Hypertension is commonly associated and acts synergistically with diabetes in increasing the risk of macrovascular and microvascular diabetic complications. Large-scale clinical trials have demonstrated that this risk is significantly reduced by intensive antihypertensive treatment, and accordingly, the Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure guideline has further lowered the blood pressure goals for diabetic subjects to <130/80 mm Hg. This implies that most diabetic patients will require the combination of two or more antihypertensive agents to achieve this blood pressure target. Although the most effective combination strategy in diabetes has not yet been determined in large-scale randomized clinical trials, a combination that includes at least one agent that interrupts the renin-angiotensin system appears to not only have a good safety profile, but may also provide additional renal and cardiovascular protection. Other antihypertensive agents should be added based on the patients risk profile and overall treatment regimen to achieve blood pressure goal. PMID- 16470080 TI - Drug-induced metabolic syndrome. AB - The metabolic syndrome is a cluster of risk factors associated with an increased risk for cardiovascular disease and type 2 diabetes. Based on data from 1988 to 1994, it is estimated that 24% of adults in the United States meet the criteria for diagnosis of the metabolic syndrome. The use of certain medications may increase the risk of the metabolic syndrome by either promoting weight gain or altering lipid or glucose metabolism. Health providers should recognize and understand the risk associated with certain medications and appropriately monitor for changes related to the metabolic syndrome. Careful attention to drug choices should be paid in patients who are overweight or have other risk factors for diabetes or cardiovascular disease. PMID- 16470081 TI - Is the development of diabetes with antihypertensive therapy a problem?--Pro. AB - Some questions about new-onset diabetes (NOD) must still be completely addressed: 1) its incidence; 2) the possible association between NOD and some classes of antihypertensive drugs; and 3) its prognostic impact. It is well known that diuretics and beta blockers can increase plasma glucose and, in available hypertension trials, diuretics and beta blockers caused a higher incidence of NOD than new antihypertensive drugs. NOD heralds a high risk of major cardiovascular events, but the absolute difference between old and new drugs was too small to significantly drive the differences in cardiovascular event rates between the two groups of treatment. This evidence suggests a judicious use of drugs more frequently associated with NOD in subjects at high risk of diabetes (impaired fasting glucose, overweight, family history of diabetes, low high-density lipoprotein cholesterol levels). The lowest effective dose of these drugs should be used, plasma glucose should be checked periodically, and concomitant lifestyle measures to prevent diabetes should be implemented with resolution. PMID- 16470082 TI - Is new-onset diabetes of clinical significance in treated hypertensive patients?- Con. AB - There is little doubt that diabetes is more common in hypertensive than normotensive individuals. There are also some data suggesting that the use of certain antihypertensive agents, i.e., diuretics and more specifically some beta blockers will increase the occurrence of new onset diabetes (NOD) when compared to other medications, especially angiotensin-converting enzyme inhibitors or angiotensin receptor blockers. The clinical significance of this 1%-3.5% difference, however, has not been established. Different studies report different outcomes. In large outcome trials the occurrence of NOD did not effect mortality or morbidity outcomes. Although one study reported that NOD had the same prognosis as pretreatment diabetes, another did not. At present, data are insufficient to suggest that NOD is of important clinical significance or that present treatment recommendations, especially regarding the use of diuretics, should be changed. PMID- 16470083 TI - Aldosterone--a hormone of cardiovascular adaptation and maladaptation. AB - Aldosterone stimulates reabsorption of sodium, sustaining blood volume and pressure in the face of salt deprivation or extracellular fluid depletion. The steroid also stimulates excretion of potassium, protecting extracellular fluid from excessive levels of that ion. These two actions are relatively rapid and clearly adaptive when appropriately initiated and terminated, but maladaptive when prolonged or excessive, causing hypertension and electrolyte imbalance. Aldosterone and other mineralocorticoids exert slower, direct effects on cells in the heart, kidneys, and vessels, leading to hypertrophy, fibrosis, and dysfunction contributing to degenerative cardiovascular diseases. The maladaptive actions of aldosterone are exacerbated by sodium chloride, angiotensin, endothelin, and certain growth factors. Damage can be minimized by antagonists of aldosterone receptors, inhibitors of the renin system, depletion of salt, and repletion of potassium and magnesium. Specific inhibitors of fibrosis and hypertrophy, and more effective inhibitors of the renin system should be useful in the future. PMID- 16470084 TI - What's in a pulse? PMID- 16470085 TI - Analysis of recent papers in hypertension. PMID- 16470086 TI - Acute and chronic oral magnesium supplementation: effects on endothelial function, exercise capacity, and quality of life in patients with symptomatic heart failure. AB - Endothelial dysfunction is an important pathophysiologic mechanism in the progression of heart failure. The objective of the present study was to determine the effects of acute and chronic oral magnesium supplementation on endothelial function in patients with symptomatic heart failure. Twenty-two symptomatic chronic heart failure patients were randomized to receive 800 mg oral magnesium oxide daily or placebo for 3 months. Data collected included large and small arterial elasticity/compliance, hemodynamic parameters, exercise capacity, and quality-of-life score at baseline, 1 week, and 3 months. Patients who received magnesium had improved small arterial compliance at 3 months from baseline compared with placebo. This study suggests that chronic supplementation with oral magnesium is well tolerated and could improve endothelial function in symptomatic heart failure patients. PMID- 16470088 TI - Peripheral edema masks the diagnoses of P pulmonale, P mitrale, and biatrial abnormality: clinical implications for patients with heart failure. AB - Peripheral edema (PED) impacts the size of the electrocardiogram, attenuating the amplitude and duration of QRS complexes and P waves. Diagnostic configurations of P pulmonale (Pp), P mitrale (Pm), and biatrial abnormality (Pb) are based on measurements of the amplitude and duration of the P waves. To investigate whether PED masks the diagnoses of Pp, Pm, and Pb, 15 patients (nine who suffered PED and six who did not gain weight during hospitalization, who served as controls) were studied. Of nine patients with PED, one had Pp, four had Pm, and four had Pb on admission, but such configurations were abolished after development of PED, except in one patient with Pm. Of six controls, one patient had Pp, three had Pm, and two had Pb, both on admission and at discharge. There was no difference in sex (p=0.61) or age (p=0.27) between the patients with PED and the controls; however, patients with PED were sicker and eventually died, while none of the controls did so. Electrocardiographic atrial abnormalities can be masked by PED, resulting from the attenuation of P waves and mediated by a decrease in the electrical impedance of the body's volume conductor due to water overload. These have clinical implications for patients with heart failure. PMID- 16470087 TI - Ambulatory hemodynamic monitoring from an implanted device: components of continuous 24-hour pressures that correlate to supine resting conditions and acute right heart catheterization. AB - Information from an implantable hemodynamic monitoring system (IHM) aids in management of patients with heart failure. This study identified which components of 24-hour IHM data best estimate resting conditions. Thirty-two patients with heart failure received an IHM in the right ventricular (RV) outflow tract. RV hemodynamics were divided into seven components of a 24-hour recording and were compared with resting supine values. Ambulatory pressures approximating rest were then compared with acute invasive catheterization values. Resting RV pressures from the IHM averaged 41+/-16/10+/-6 mm Hg and estimated pulmonary artery diastolic pressure was 21+/-8 mm Hg. Nighttime (midnight to 4 a.m.) minimum pressures from the IHM best approximated supine resting conditions. RV and pulmonary artery pressures during catheterization were higher than the nighttime minimum, although RV diastolic pressure was not statistically different. Minimum RV and pulmonary artery pressures during nighttime approximate observed resting conditions; invasive catheterization pressures are higher than IHM resting values. PMID- 16470089 TI - Assigning patients with heart failure to observation status: B-type natriuretic peptide, ejection fraction, or physician judgment. AB - It is of clinical and financial importance to identify those heart failure patients who are likely to improve rapidly. The authors evaluated, as predictors of short-term resolution, three clinical variables often used to predict long term outcome. Consenting patients admitted to the emergency department with dyspnea were examined daily until resolution (symptom reversion to baseline absent worsening clinical signs or x-ray). The authors then compared hours to resolution of heart failure with serum B-type natriuretic peptide (BNP), ejection fraction (EF), and admitting physician prediction. They calculated negative and positive predictive values for EF, BNP, and physician opinion using quintile and published severity standards as cutoffs (for EF and BNP). Among 85 patients, BNP <400 pg/mL and <1000 pg/mL had poor positive predictive values (34% and 22%, respectively), as did EF (21%) and physician opinion (16%-21%, depending on physician confidence). Combining tests did not improve prediction. These results do not support the use of BNP, EF, or clinical assessment in triaging heart failure patients to short-stay status. PMID- 16470091 TI - Managing the post-left ventricular assist device patient. AB - The implantation of ventricular assist devices allows the opportunity for patients with intractable heart failure to have improved quality and quantity of life. The devices may be implanted after failed attempts to wean from bypass, as a bridge to transplantation, or as destination therapy. Key issues following the implantation of assist devices include the prevention of right ventricular failure, appropriate pharmacologic management, prevention and management of infection, and detection and treatment of device dysfunction. PMID- 16470092 TI - Reversible attenuation of the ECG voltage due to peripheral edema associated with treatment with a COX-2 inhibitor. AB - A 74-year-old man developed peripheral edema as a side effect of the cyclooxygense-2 selective receptor inhibitor rofecoxib, which he had been taking for severe chronic arthritis. Discontinuation of rofecoxib led to augmentation of electrocardiographic (ECG) voltage and loss of weight gain (and reversibility of peripheral edema), which correlated well (r=0.82; p=0.0002). Other good correlations of the weight and other ECG variables and intercorrelations of ECG parameters underscore the multiple reversible influences peripheral edema has on the ECG. This case highlights an enhanced role of the ECG in monitoring patient therapy with other than strictly cardiovascular drugs. Recently, a syndrome pertaining to the influence of peripheral edema on the ECG was described; its mechanism is via the transforming effect of the body volume conductor on the surface transfer of the heart's potentials. The objective of this report is to describe a patient who developed peripheral edema as a side effect of a cyclooxygenase-2 selective receptor inhibitor. PMID- 16470090 TI - Heart failure, myocardial stunning, and troponin: a key regulator of the cardiac myofilament. AB - This review discusses post-translational modifications of myofilament regulatory proteins, particularly troponin, associated with heart failure and myocardial stunning--two common disease processes. Altered phosphorylation, partial proteolysis and, possibly, oxidative damage to myofilament proteins may result in abnormalities in both systolic and diastolic function. At a molecular level, these changes may lead to abnormalities in crossbridge cycling and tension development and result in inefficiencies in utilization of energy. Understanding these alterations may lead to new targeted therapies. PMID- 16470094 TI - Historical vignettes in heart failure. PMID- 16470096 TI - What happens to the public interest when medical care becomes a business? PMID- 16470097 TI - Saving lives and saving money in the 21st century. PMID- 16470095 TI - Hyponatremia and heart failure--treatment considerations. AB - Hyponatremia as it occurs in the heart failure patient is a multifactorial process. The presence of hyponatremia in the heart failure patient correlates with both the severity of the disease and its ultimate outcome. The therapeutic approach to the treatment of hyponatremia in heart failure has traditionally relied on attempts to improve cardiac function while at the same time limiting fluid intake. In more select circumstances, hypertonic saline, loop diuretics, and/or lithium or demeclocycline have been used. The latter two compounds act by retarding the antidiuretic effect of vasopressin but carry with their use the risk of serious renal and/or cardiovascular side effects. Alternatively, agents that selectively block the type 2 vasopressin receptor increase free water excretion without any of the adverse consequences of other therapies. Conivaptan, lixivaptan, and tolvaptan are three such aquaretic drugs. Vasopressin receptor antagonists will redefine the treatment of heart failure-related hyponatremia and may possibly evolve as adjunct therapies to loop diuretics in diuretic-resistant patients. PMID- 16470098 TI - Can we cross the quality chasm? The case for realistic optimism. PMID- 16470099 TI - The clinical examination is still the lynchpin of quality cardiovascular care. PMID- 16470102 TI - Pay for performance: challenges and opportunities. PMID- 16470100 TI - The inevitability of insurance disputes, and tips to ensure health insurance providers pay. PMID- 16470101 TI - Pay for performance: health care revived or health care imbibed? PMID- 16470103 TI - Medicare health support: reinventing chronic care. AB - Medicare Health Support is a groundbreaking new initiative to help chronically ill Medicare beneficiaries improve their health and quality of life. If the ongoing pilot programs are successful, they may have far-reaching effects on population health and patient care. PMID- 16470105 TI - Personal reflections on coronary artery calcium quantitation by CT. PMID- 16470104 TI - Coronary calcium scanning. AB - While there is no doubt that high-risk patients (those with more than a 20% 10 year risk of a future cardiovascular event) need more aggressive preventive therapy, a majority of cardiovascular events occur in individuals at intermediate risk (10%-20% 10-year risk). Data suggest that it will be most cost-effective to concentrate screening efforts on this group of patients. Coronary artery calcium has been shown to be highly specific for atherosclerosis, occurring only in the intima of the coronary arteries. There is evidence to show that elevated coronary calcium scores are predictive of cardiovascular events, both independently of and incrementally to conventional cardiovascular risk factors. Based on current available data, patients with increased plaque burdens (increased coronary calcium scores) are approximately 10 times more likely to suffer a cardiac event over the next 3-5 years. Coronary calcium scores have outperformed conventional risk factors, high sensitivity C-reactive protein, and carotid intima-media thickness as a predictor of cardiovascular events. Both electron beam tomography and multidetector computed tomography can accurately detect and quantify the coronary calcium scores. In summary, coronary calcium detection significantly improves the accuracy of global cardiovascular risk prevention, the noninvasive tracking of the atherosclerotic burden, and the prediction of cardiovascular events. PMID- 16470107 TI - William Osler: slow pulse, stokes-adams disease, and sudden death in families. AB - In 1903, William Osler, then at Johns Hopkins University, published "On the So Called Stokes-Adams Disease (Slow Pulse with Syncopal Attacks, etc.)" in The Lancet, classifying a syndrome in evolution. There are thinly disguised references to a brother and to himself in the article, suggesting that Osler was concerned about a family and personal predisposition. Osler's decision to move to Oxford was triggered in part by his personal concerns about cardiac disease. Then, in 1909, Osler contributed a chapter on Stokes-Adams disease to Allbutt and Rolleston's A System of Medicine, complemented by a brilliant pathologic section by the renowned anatomist-morphologist, Arthur Keith. Osler's original contributions involved his emphasis on the importance of family history, his careful clinical and natural history observations, and his recognition of the familial occurrence of bradycardia, Stokes-Adams disease, sudden death, and cardiomyopathy. PMID- 16470106 TI - Evolution and scaling of atrioventricular conduction time in mammals: part 1. AB - Scaling can be defined as the adjustment of a structure, a function, or an organ to the size of the mammalian body. An example is the size of the heart in relation to the size of the body. The duration of the PR interval on the electrocardiogram (atrioventricular delay) in relation to the size of the heart is a perplexing example of scaling. During evolution, mammalian species changed their shape, size, and function while adapting to the habitat in which they had to live and survive. This review deals with the problem of the apparent mismatch in scaling of the atrioventricular delay (PR interval) in relation to the size of the mammalian heart from mouse to whale. PMID- 16470108 TI - Trouble breathing: use of echocardiography to define mechanism(s) and guide treatment. PMID- 16470109 TI - Sequential myocardial perfusion imaging and cardiac CT: what to do with incidental CT findings? PMID- 16470110 TI - CT angiography: a case from the Wisconsin Heart Hospital. PMID- 16470111 TI - Atrial (? sinus) tachycardia with block (well concealed). PMID- 16470112 TI - The maturing immune system: implications for development and testing HIV-1 vaccines for children and adolescents. PMID- 16470113 TI - Fluorodeoxyglucose imaging in healthy subjects with HIV infection: impact of disease stage and therapy on pattern of nodal activation. AB - OBJECTIVES: Nodal uptake in areas of lymphocyte activation can be visualized using fluorodeoxyglucose (FDG). Various patterns of FDG accumulation in HIV positive subjects have been described previously and hypothesized to potentially represent regions of active HIV replication and or nodal activation. We evaluated the utility of FDG scanning as a tool to study HIV pathogenesis. DESIGN: We evaluated FDG biodistribution visually and quantitatively in HIV-negative individuals and various groups of HIV-infected subjects to determine the impact on pattern of nodal activation of: HIV infection; stage of HIV infection and degree of viremia; and HAART. In addition, we attempted to image anatomical site(s) of on-going HIV replication in subjects with suppressed HIV viremia on ART, but who subsequently discontinued ART. METHOD: We performed FDG imaging on five groups: HIV-negative, HIV-positive with early infection, HIV-positive with advanced disease, HIV-positive with suppressed viral loads, and HIV-positive who stopped ART. RESULTS: Healthy HIV subjects with suppressed viral loads and HIV negative individuals had no or little FDG nodal accumulation or any other hypermetabolic areas, whereas viremic subjects with early and advanced HIV had increased FDG in peripheral nodes, indicating that FDG potentially identifies areas of HIV replication. FDG biodistribution was similar between early and advanced-stage. Four of five subjects taken off ART had negative baseline scans but developed nodal uptake and increases in viral load. CONCLUSIONS: Abnormal FDG accumulation occurs in nodes of subjects with detectable viral loads. Interruption of effective ART results in activation of previously quiescent nodal areas. PMID- 16470114 TI - Broad neutralization and complement-mediated lysis of HIV-1 by PEHRG214, a novel caprine anti-HIV-1 polyclonal antibody. AB - OBJECTIVES: To assess the potency, breadth of action, and mechanism of action of the polyclonal goat anti-HIV antibody, PEHRG214. DESIGN: Typical human antibody responses to HIV-1 infection are unable to neutralize virus efficiently, clear the infection, or prevent disease progression. However, more potent neutralizing antibodies may be capable of playing a pivotal role in controlling HIV replication in vivo. PEHRG214 is a polyclonal caprine antibody raised against purified HIV-associated proteins, such that epitopes that are immunologically silent in humans may potentially be recognized in another species. It has been administered safely to HIV-infected individuals in Phase I clinical trials. METHODS: The anti-HIV activity of PEHRG214 was assessed using neutralization and virion lysis assays. The target proteins for PEHRG214 activity were investigated using flow cytometry and by adsorption of anti-cell antibodies from the antibody cocktail. RESULTS: PEHRG214 strongly neutralized a diverse range of primary HIV-1 isolates, encompassing subtypes A to E and both CCR5 and CXCR4 phenotypes. Neutralization was enhanced by the presence of complement. PEHRG214 also induced complement-mediated lysis of all HIV-1 isolates tested, and recognized or cross reacted with a number of host cell proteins. Lysis was abrogated by adsorption with T and/or B cells expressing GPI-linked proteins, but not by GPI-deficient B cells or red blood cells. CONCLUSIONS: PEHRG214 was found to potently neutralize and lyse HIV-1 particles. By targeting host cell proteins present in the viral envelope, which are conserved among all strains tested, PEHRG214 potentially opens up a highly novel means of eliminating circulating virus in infected individuals. PMID- 16470115 TI - Viral dynamics after starting first-line HAART in HIV-1-infected children. AB - BACKGROUND: After starting HAART, the plasma HIV-1 RNA (pVL) declines rapidly to undetectable levels in most treated adults and children. The viral dynamics in children are assumed to differ from those in adults. Therefore viral decay and time to reach a pVL of < 400 copies/ml during the first weeks after starting HAART were studied in a cohort of HIV-1-infected children. METHODS: Viral decay expressed as half-life and time to reach a pVL of < 400 copies/ml in 39 HIV-1 infected children starting HAART were calculated and correlated with age, pretreatment with antiretroviral mono- or duo-therapy, and baseline pVL. RESULTS: Baseline pVL correlated with age (r, -0.41; P = 0.01). Median half-life of the virus was 2.1 days (interquartile range, 1.8-3.0 days). No correlation was found between the half-life of the virus and the baseline pVL at the start of treatment, antiretroviral pretreatment or age. Eight children did not reach a pVL of < 400 copies/ml with the first allocated medication regimen. These children were significantly younger than those in whom HIV was successfully suppressed (P = 0.009). The remaining 31 children reached a pVL of < 400 copies/ml in a median of 8.1 weeks after the start of therapy; time to reach a pVL of < 400 copies/ml was only correlated with baseline pVL. CONCLUSIONS: These results suggest that pVL at baseline correlated with age. HAART was able to suppress pVL below the lower limit of detection in children with a viral decay rate of 2.1 days, similar to adults and irrespective of baseline pVL. PMID- 16470116 TI - Hepatic steatosis in HIV-HCV coinfected patients: analysis of risk factors. AB - OBJECTIVE: To evaluate the prevalence and severity of steatosis and possible interactions between steatosis, host factors, viral factors, and treatment for HIV infection in HIV-hepatitis C virus (HCV) coinfected patients. METHODS: Steatosis was assessed among 395 HIV-HCV coinfected patients who were enrolled in the ANRS trial HC02 Ribavic and for whom histological data were available. Steatosis was graded as follows: 0 (none); 1 (< 30% hepatocytes containing fat); 2 (30-70%); 3 (> 70%). RESULTS: Steatosis was present in 241 patients (61%), of whom 149 (38%) had grade 1, 64 (16%) grade 2 and 28 (7%) grade 3. In multivariate analysis, the following five independent risk factors were associated with steatosis: HCV genotype 3 [odds ratio (OR), 3.02; 95% confidence interval (CI), 1.91-4.79; P < 0.0001], the mean METAVIR fibrosis score (OR, 1.43; 95% CI, 1.11 1.84; P = 0.0053), the body mass index (BMI; OR, 1.13; 95% CI, 1.05-1.21; P = 0.0013), HCV viral load (OR. 1.65; 95% CI, 1.22-2.23; P = 0.0012) and ferritin (OR, 1.13; 95% CI, 1.06-1.21; P < 0.0003). As HCV genotype 3 was a risk factor for steatosis, further exploratory analyses were stratified according to the HCV genotype (1 and 3). Factors independently associated with steatosis were BMI and HCV viral load in patients with HCV genotype 3 infection and the mean METAVIR fibrosis score, the BMI and ferritin in patients with HCV genotype 1 infection. CONCLUSION: Steatosis is particularly frequent in HIV-HCV coinfected patients, who appear to have the same risk factors for steatosis as HCV monoinfected patients. None of the characteristics of HIV infection, including antiretroviral therapy, was independently associated with steatosis. PMID- 16470117 TI - Differential susceptibility of human thymic dendritic cell subsets to X4 and R5 HIV-1 infection. AB - OBJECTIVES: Human thymus can be infected by HIV-1 with potential consequences on immune regeneration and homeostasis. We previously showed that CD4 thymocytes preferentially replicate CXCR4 tropic (X4) HIV-1 dependently on interleukin (IL) 7. Here we addressed the susceptibility of thymic dendritic cells (DC) to HIV-1 infection. METHODS: We investigated the replication ability of CXCR4 or CCR5 (R5) tropic HIV-1 in thymic micro-explants as well as in isolated thymic CD11clowCD14- DC, CD11chighCD14+ DC and plasmacytoid DC subsets. RESULTS: Thymic tissue was productively infected by both X4 and R5 viruses. However, X4 but not R5 HIV-1 replication was enhanced by IL-7 in thymic micro-explants, suggesting that R5 virus replication occurred in cells other than thymocytes. Indeed, we found that R5 HIV-1 replicated efficiently in DC isolated from thymic tissue. The replicative capacity of X4 and R5 viruses differed according to the different DC subsets. R5 but not X4 HIV-1 efficiently replicated in CD11chighCD14+ DC. In contrast, no HIV-1 replication was detected in CD11clowCD14- DC. Both X4 and R5 viruses efficiently replicated in plasmacytoid DC, which secreted interferon alpha upon HIV-1 exposure. Productive HIV-1 infection also caused DC loss, consistent with different permissivity of each DC subset. CONCLUSIONS: Thymic DC sustain high levels of HIV-1 replication. DC might thus be the first target for R5 HIV-1 infection of thymus, acting as a Trojan horse for HIV-1 spread to thymocytes. Furthermore, DC death induced by HIV-1 infection may affect thymopoiesis. PMID- 16470118 TI - Safety and tolerability of tenofovir vaginal gel in abstinent and sexually active HIV-infected and uninfected women. AB - OBJECTIVES: To establish the highest practical dose and frequency (HPDF) of 0.3% or 1% tenofovir vaginal gel applied once or twice daily by sexually abstinent HIV uninfected women, and to evaluate the safety, tolerability and systemic pharmacokinetics of the HPDF in abstinent and sexually active HIV-negative and HIV-infected women. METHODS: Eighty-four women, enrolled in sequential cohorts, used the study product for 14 consecutive intermenstrual days. Safety laboratory assessments and pelvic examinations were carried out during five study visits, with colposcopy at enrollment and on day 14. Samples for pharmacokinetics were collected before and after the initial tenofovir gel use and at day 13. RESULTS: The 1% tenofovir gel used twice daily was as well tolerated as other regimens used by the 48 HIV-negative sexually abstinent women, establishing the HPDF. Although 92% of the women reported at least one adverse event, the majority were mild (87%) and involved the genitourinary tract (70%). One possibly product related severe adverse event involving lower abdominal cramping was reported by a sexually abstinent woman who used 0.3% gel twice daily. Serum tenofovir levels were low but detectable in 14 of the 25 women. No new HIV RNA resistance mutations were detected after 2 weeks of tenofovir gel in the 24 HIV-infected participants. No significant systemic toxicity was detected. CONCLUSION: A 2-week course of 1% tenofovir vaginal gel used twice daily was well tolerated in sexually abstinent and sexually active HIV-negative and HIV-positive women. Systemic tenofovir absorption occurred. Expanded safety and effectiveness testing is warranted. PMID- 16470119 TI - Impact of pregnancy on abacavir pharmacokinetics. AB - OBJECTIVE: To describe abacavir pharmacokinetics during pregnancy and postpartum; physiological changes during pregnancy are known to affect antiretroviral drug disposition. DESIGN: The Pediatric AIDS Clinical Trials Group P1026s study is an on-going, prospective, non-blinded pharmacokinetic study of pregnant women receiving one or more antiretroviral drugs for routine clinical care, including a cohort receiving abacavir 300 mg twice daily. METHODS: Serial plasma samples (predose, 1, 2, 4, and 6 h postdose) obtained antepartum (30-36 weeks of gestation) and again postpartum (6-12 weeks after delivery) were assayed for abacavir concentration by reversed-phase high-performance liquid chromatography. RESULTS: Antepartum evaluations were available for 25 women [mean age, 28.6 years (SD, 6); mean third-trimester weight 92 kg (SD, 35.4); and race/ethnicity 52% black, 28% Hispanic, 16% white, 4% Asian], with geometric mean abacavir area under the concentration-time curve (AUC) of 5.9 mg.h/l [90% confidence interval (CI), 5.2-6.8] and maximum plasma concentration (Cmax) of 1.9 mg/l (90% CI, 1.6 2.2). Seventeen women completed postpartum sampling, and the ratios of antepartum to postpartum AUC and Cmax were 1.04 (90% CI, 0.91-1.18) and 0.79 (90% CI, 0.65 0.98), respectively. CONCLUSIONS: Abacavir AUC during pregnancy was similar to that at 6-12 weeks postpartum and to that for non-pregnant historical controls (5.8 mg.h/l). Consequently, pregnancy does not appear to affect overall abacavir exposure significantly or to necessitate dose adjustments. PMID- 16470120 TI - Acute renal failure in hospitalized patients with HIV: risk factors and impact on in-hospital mortality. AB - BACKGROUND: Kidney disease is an increasingly important complication of HIV. OBJECTIVES: To examine the incidence and predictors of acute renal failure before and after the introduction of HAART, and the impact of acute renal failure on in hospital mortality in the post-HAART era. METHODS: Adults hospitalized in acute care hospitals in New York State during 1995 (pre-HAART) or 2003 (post-HAART) were identified from the state Planning and Research Cooperative System database. HIV status was defined by primary or secondary diagnosis code. The impact of HIV and HAART on the incidence of acute renal failure and mortality, and the impact of acute renal failure on mortality, was assessed using chi analysis and multivariate regression. RESULTS: There were 52,580 HIV-infected patients discharged from hospital in 1995 and 25,114 in 2003. Compared with uninfected patients, HIV-infected patients had an increased incidence of acute renal failure in both the pre-HAART [adjusted odds ratio (OR), 4.62; 95% confidence interval (CI), 4.30-4.95] and post-HAART eras (adjusted OR, 2.82; 95% CI, 2.66-2.99). In the post-HAART cohort, acute renal failure was associated with traditional predictors such as age, diabetes mellitus, and chronic kidney disease, as well as acute or chronic liver failure or hepatitis coinfection (P < 0.001 for all comparisons). Acute renal failure was associated with mortality among HIV infected patients in the post-HAART era (OR, 5.83; 95% CI, 5.11-6.65). CONCLUSIONS: Acute renal failure remains common among hospitalized patients with HIV and is associated with chronic kidney disease, liver disease, and increased mortality. PMID- 16470121 TI - Vpr and HIV-1 disease progression: R77Q mutation is associated with long-term control of HIV-1 infection in different groups of patients. AB - BACKGROUND: Vpr (viral protein R) is a 96 amino acids soluble protein that is expressed late during viral replication. Recent studies have focused on the role of a mutation at position 77 that might be associated with the condition of long term non-progression, but data are still controversial. PATIENTS AND METHODS: Fifteen long-term non-progressors (LTNP), 19 therapy-naive HIV-1-infected patients with progressive disease (Pr), 23 HIV-1-infected patients receiving sub optimal therapy with dual nucleoside [nucleoside reverse transcriptase inhibitor (NRTI)] therapy but efficiently controlling viral replication (STP) and 19 antiretroviral therapy multi-experienced patients with actively replicating virus (MEP) were analysed. HIV-RNA was extracted from plasma samples, the Vpr region was amplified, cloned and sequenced. The Pol gene was amplified, directly sequenced and analysed using Sequence Navigator software. RESULTS: A significantly higher prevalence of the R77Q mutation was evidenced both in LTNP (86.7%) and STP (73.9%) in comparison with Pr (42.1%) and MEP (42.1%), (P = 0.007). Comparing groups of patients with progressive disease (Pr + MEP) and groups with non-progressive disease (LTNP + STP) the probability of harbouring the R77Q mutation was significantly higher in non-progressors (odds ratio, 5.16; P = 0.001). CONCLUSIONS: Our results support the hypothesis of the association of R77Q mutation in the Vpr gene with delayed progression of HIV-1 disease. R77Q does not seem to be linked to a particular viral strain but might be associated to immunologic selection. The R77Q mutation might reduce CD4+ T-cell depletion possibly affecting T-cell survival in vivo by altering the pro-apoptotic activity of Vpr. PMID- 16470122 TI - Effect of hepatitis C coinfection on discontinuation and modification of initial HAART in primary HIV care. AB - OBJECTIVES: To estimate the effect of hepatitis C (HCV) coinfection on time to first occurrence of either discontinuation or modification of initial HAART among previously antiretroviral therapy-naive HIV-infected patients. METHODS: The analysis included antiretroviral therapy-naive patients who initiated HAART prior to November 2003 and were participating in the University of North Carolina Center for AIDS Research, HIV/AIDS observational clinical cohort. The effect of HCV status on time to first occurrence of either HAART discontinuation or modification was assessed using Kaplan-Meier survival estimates and multivariable proportional hazards regression was used to estimate hazard ratios. RESULTS: Of 296 patients initiating HAART, 22% were coinfected with HCV. During a median follow-up of 473 days [interquartile range (IQR), 167-940] from HAART initiation, 104 (35%) patients discontinued and 91 (31%) modified their first regimen. Reasons for discontinuation and modification were comparable by HCV serostatus and included treatment failure (12%), toxicity (41%), and barriers to adherence (47%). The median time to first occurrence of either discontinuation or modification among HCV-infected patients was 401 days (IQR, 128-821), and among HCV-uninfected patients was 493 days (IQR, 204-952) (P = 0.22). After adjustment for baseline demographic and clinical characteristics, the hazard ratio contrasting HCV-infected with HCV-uninfected patients was 1.39 (95% confidence interval, 0.95-2.03; P = 0.09). CONCLUSION: HCV coinfection was only marginally associated with a shorter duration of an initial HAART regimen, suggesting optimization of a first HAART regimen may not appreciably depend on HCV serostatus. PMID- 16470123 TI - Correlates of HIV infection among former blood/plasma donors in rural China. AB - BACKGROUND: In 1995, when the first cases of HIV infection were reported among former plasma donors (FPDs), the Chinese government closed all commercial plasma collection stations. OBJECTIVE: To determine the prevalence of HIV among FPDs and non-donors in affected villages in Anhui, China. METHODS: A cross-sectional survey was conducted among residents, aged 25-55 years, in 40 villages randomly selected from villages with many former blood/plasma donors, using a two-stage clustered sampling method. A questionnaire was administered face-to-face to 1997 villagers without collecting any identifying information, and venous blood specimens were collected for HIV testing with two enzyme-linked immunosorbent assays and western blotting. EpiData was used for data entry, and STATA was used for data analysis. RESULTS: Overall HIV prevalence was 10.8%, with values of 15.1% among FPDs and 4.8% among non-donors. Among FPDs, factors associated with HIV infection included: donating plasma more than 10 times [odds ratio (OR) 4.09; P < 0.001] compared with subjects who donated 1-3 times; spouse being HIV positive (OR, 4.06; P = 0.001); and being male (OR, 2.04; P = 0.011). Condom use was rare, and was not associated with HIV infection (OR, 1.09; P = 0.872). Among non-plasma donors, spouse being HIV-positive (OR, 11.07, P < 0.001) and having multiple sexual partners (OR, 7.04; P = 0.006) were associated with HIV infection. CONCLUSIONS: The prevalence of HIV infection is high among rural residents in villages with former commercial plasma businesses. Plasma but not blood donations were associated with HIV infection. The HIV/AIDS epidemic has spread to non-donors primarily through sexual transmission. HIV/AIDS education, testing, and condoms should be promoted urgently to prevent further transmission. PMID- 16470124 TI - Association between genital schistosomiasis and HIV in rural Zimbabwean women. AB - OBJECTIVE: To determine the association between female genital Schistosoma haematobium infection and HIV. DESIGN AND METHODS: A cross-sectional study with a 1-year follow-up. Gynecological and laboratory investigations were performed for S. haematobium and HIV. Sexually transmitted infections, demographic and urogenital history were analysed as confounders. The participants were 527 sexually active, non-pregnant, non-menopausal women between the ages of 20 and 49 years. The setting was a rural Zimbabwean community where S. haematobium related lesions were found in 46% of the women, HIV in 29% and herpes simplex type- 2 (HSV-2) in 65%. RESULTS: In permanent residents (>3 years residency), HIV was found in 41% (29/70) of women with laboratory proven genital schistosomiasis as opposed to 26% HIV positive (96/375) in the schistosomal ova negative group [odds ratio (OR), 2.1; 95% confidence interval (CI), 1.2-3.5; P = 0.008. In multivariate analysis S. haematobium infection of the genital mucosa was significantly associated with HIV seropositivity (adjusted OR, 2.9; 95% CI, 1.11 7.5; P = 0.030). All seven women who became HIV positive during the study period (seroincidence 3.1%) had signs of S. haematobium at baseline. In accordance with other studies HIV was significantly associated with HSV-2 (OR, 3.0; 95% CI, 1.7 5.3; P < 0.001), syphilis and human papillomavirus. The highest HIV prevalence (45%) was found in the 25-29 years age group. CONCLUSION: Women with genital schistosomiasis had an almost three-fold risk of having HIV in this rural Zimbabwean community. Prospective studies are needed to confirm the association. PMID- 16470125 TI - Mobility and HIV in Tanzanian couples: both mobile persons and their partners show increased risk. AB - OBJECTIVE: To investigate how mobility is related to sexual risk behavior and HIV infection, with special reference to the partners who stay behind in mobile couples. METHODS: HIV status, sexual behavior and demographic data of 2800 couples were collected in a longitudinal study in Kisesa, rural Tanzania. People were considered short-term mobile if they had slept outside the household at least once on the night before one of the five demographic interviews, and long term mobile if they were living elsewhere at least once at the time of a demographic round. RESULTS: Overall, whereas long-term mobile men did not report more risk behavior than resident men, short-term mobile men reported having multiple sex partners in the last year significantly more often. In contrast, long-term mobile women reported having multiple sex partners more often than resident women (6.8 versus 2.4%; P = 0.001), and also had a higher HIV prevalence (7.7 versus 2.7%; P = 0.02). In couples, men and women who were resident and had a long-term mobile partner both reported more sexual risk behavior and also showed higher HIV prevalence than people with resident/short-term mobile partners. Remarkably, risk behavior of men increased more when their wives moved than when they were mobile themselves. CONCLUSIONS: More sexual risk behavior and an increased risk of HIV infection were seen not only in mobile persons, but also in partners staying behind. Interventions aiming at reducing risk behavior due to mobility should therefore include partners staying behind. PMID- 16470126 TI - Decline in sexually transmitted infection prevalence and HIV incidence in female barworkers attending prevention and care services in Mbeya Region, Tanzania. AB - OBJECTIVE: To assess trends in sexually transmitted infection (STI) prevalence and HIV incidence and associated factors in a cohort of female barworkers exposed to behavioural interventions and STI screening and treatment. METHODS: An open cohort of 600 female barworkers in Mbeya Region, Tanzania was offered 3-monthly information and education sessions on HIV/STI and reproductive health, voluntary HIV counselling and testing and clinical health check-ups including STI syndromic management with simple STI laboratory support. Outcome assessments included HIV, herpes simplex virus type 2 (HSV-2) and syphilis serology, polymerase chain reaction for Neisseria gonorrhoeae, Chlamydia trachomatis and ulcerative STI, microscopy for Trichomonas vaginalis, Candida albicans and bacterial vaginosis and interviews on sociodemographic and behavioural characteristics. RESULTS: Over a period of 30 months 600 barworkers were enrolled at the baseline examination round and 153 thereafter as replacements for losses to follow-up. At 3-monthly examinations the prevalence of gonorrhoea declined steadily from 22.2 to 6.8% (odds ratio for trend per quarter: 0.81; P < 0.001). The prevalence of all other STI/RTI, except for genital herpes and bacterial vaginosis, also decreased significantly. HIV incidence declined from 13.9/100 to 5.0/100 person-years over three consecutive 9-month periods. HIV incidence was significantly associated with genital ulcers and positive syphilis serology, but not with genital herpes or HSV-2 seropositivity. CONCLUSION: A relatively simple intervention consisting of regular 3-monthly STI screening and syndromic management in combination with HIV/STI information and counselling sessions was well accepted and effective in reducing STI among barworkers. Such interventions should be implemented more widely in high-risk environments in sub-Saharan Africa. PMID- 16470127 TI - Immune-based interventions in HIV infection: doing the right studies, getting the right answers. PMID- 16470128 TI - The effect of atorvastatin treatment on HIV-1-infected patients interrupting antiretroviral therapy. AB - We conducted a pilot study to assess the effect of atorvastatin on HIV replication. Patients with stable HAART-controlled infection interrupted therapy and were randomly assigned to a control group or to start atorvastatin 40 or 80 mg/day. Statin groups showed lower serum cholesterol but similar viral loads and CD4 T-cell counts to the control group at weeks 4 and 12. Paradoxically, baseline serum cholesterol, but not atorvastatin, influenced viral rebound at week 4. PMID- 16470129 TI - Dual tropism of HIV-1 envelopes derived from renal tubular epithelial cells of patients with HIV-associated nephropathy. AB - The phenotype of HIV-1 gp120 envelope derived from renal epithelium and peripheral blood mononuclear cells (PBMC) of patients with HIV-associated nephropathy was investigated in vitro. Chimeric viruses were derived from kidney or blood and used to infect primary CD4+T cells, cell lines expressing single co receptors and a renal epithelial cell line HPT-1. HIV-1 variants derived from renal epithelium were dual tropic whereas simultaneously derived viruses from PBMC were R5-tropic. Utilization of alternative co-receptors CCR3, BONZO and BOB, also differed. PMID- 16470130 TI - Variability in the P6gag domains of HIV-1 involved in viral budding. AB - The genetic variability within PT/SAP, LYP and LXXLF HIV-1 P6gag motifs, required for the binding to Tsg101 and AIP1 cellular host proteins during viral budding, was examined in 122 HIV-infected subjects. PT/SAP duplications were statistically more frequent in B versus non-B subtypes. Substitutions at LYP were fourfold less frequent in antiretroviral-experienced only in clade B. P6gag variability across HIV-1 subtypes and after antiretroviral exposure may influence interactions with host cells involved in viral budding. PMID- 16470131 TI - Long-term HIV-specific responses and delayed resumption of antiretroviral therapy after peptide immunization targeting dendritic cells. AB - Long-term HIV-specific immune responses and clinical outcomes were evaluated in HIV-infected patients previously immunized with p24-like peptides (Vacc-4x) targeting dendritic cells (DC). Vacc-4x-induced cellular immune responses were unchanged 1.5 years after completing immunization, and 62% were still off combined antiretroviral treatment (CART). The magnitude of early Vacc-4x responses determined whether the resumption of CART was clinically indicated 2 years after enrollment. These observations encourage further exploration of both Vacc-4x and other HIV peptide-based immunization regimens targeting DC. PMID- 16470132 TI - Human bites: a rare risk factor for HIV transmission. PMID- 16470133 TI - Cidofovir treatment of HIV-associated cytomegalovirus polyradiculopathy. PMID- 16470135 TI - Pakistan/India open borders ... to HIV? PMID- 16470134 TI - Concerns regarding a randomized trial of two postexposure prophylaxis regimens. PMID- 16470136 TI - HIV-discordant couples and parenthood: how are we dealing with the risk of transmission? PMID- 16470138 TI - The 'vices' in devices. PMID- 16470139 TI - A futuristic perspective on clinical studies of cardiac resynchronization therapy for heart failure patients. AB - PURPOSE OF REVIEW: Heart failure is a major public health problem. Many heart failure patients have electrical and mechanical ventricular dyssynchrony, which are risk factors for death in heart failure patients. RECENT FINDINGS: Cardiac resynchronization therapy, by stimulating both ventricles, is a strategy to improve ventricular dyssynchrony. SUMMARY: This paper describes the historic development of this therapy; reviews the results of completed clinical cardiac resynchronization therapy studies, and discusses ongoing and future studies. PMID- 16470140 TI - Pacing delivered rate and rhythm control for atrial fibrillation. AB - PURPOSE OF REVIEW: Device therapy for atrial fibrillation remains contentious despite the recognized benefit of atrial pacing in sinus node dysfunction. There are various new specialized pacing algorithms that aim to provide rhythm or rate control in atrial fibrillation. We review the various options for device therapy and the evidence available concerning their effectiveness. RECENT FINDINGS: Randomized trials on preventative algorithms for atrial fibrillation have not shown consistent benefit. Anti-tachycardia pacing for atrial fibrillation has inherent problems illustrated in this review and has failed to demonstrate objective improvement except in the case of atrial flutter. Several large randomized trials have demonstrated an adverse outcome with right ventricular apical pacing. These studies have shown an increase in atrial fibrillation with ventricular pacing. Recent studies have emphasised the importance of right ventricular apical pacing in burden of atrial fibrillation and therefore we discuss the likely confounding effect on previous trials and speculate on future directions. SUMMARY: The use of a device with atrial fibrillation prevention algorithms in a patient with a bradycardia indication for pacing is not unreasonable but there is no hard evidence of benefit. Patients with sinus node dysfunction should be paced in the atrium alone. There is no indication for use of a device for atrial fibrillation without a conventional indication for pacing. PMID- 16470141 TI - Rate control in atrial fibrillation: looking beyond the average heart rate. AB - PURPOSE OF REVIEW: The aim of this article is to provide a perspective on rate control in atrial fibrillation which emphasizes patient wellbeing (exercise tolerance, symptoms, quality of life) over attempts to reduce resting or exercise heart rate to an arbitrary range. RECENT FINDINGS: Recent trials of rhythm versus rate control strategies of treatment in patients with atrial fibrillation suggest that rate control is a viable first line strategy in many patients. The adverse consequences of atrial fibrillation with rapid ventricular response are partly due to factors other than rate itself, such as irregularity of ventricular response, and variable changes in autonomic nervous system output. Digoxin, calcium channel blockers, and beta-blockers cause a similar reduction in resting heart rate. Beta blockers are the most potent at reducing exercise heart rate, followed by calcium channel blockers and digoxin. Exercise tolerance is occasionally improved by digoxin, sometimes improved by calcium channel blockers and not improved by (and sometimes decreased by) beta-blockers. Information about quality of life with different rate control regimens is sparse. SUMMARY: Rate control in atrial fibrillation provides important benefits to patients in terms of symptoms, quality of life and prevention of late consequences of uncontrolled rate (such as tachycardia induced cardiomyopathy). Restricting treatment objectives to achievement of a specific heart rate range on resting or exercise electrocardiogram may result in lack of patient benefit or worsened symptoms. Understanding the nuances of rate control when treating individual patients and interpreting existing evidence allows patients to experience the most benefit from this treatment strategy. PMID- 16470143 TI - Congenital mitral valve surgery: techniques and results. AB - PURPOSE OF REVIEW: Congenital lesions of the mitral valve are rare. Conservative surgery is recognized as the best option. In complex anatomy, however, replacement is the only solution to achieve an acceptable result. This review aims to study the long-term follow-up of classical treatments, conservative or replacement, and to examine new technical advances. RECENT FINDINGS: The long term results of conservative surgery are confirmed with a low incidence of reoperation except in mitral valve stenosis. The Ross II operation using a pulmonary autograft is a difficult technique that may be useful in the youngest patient group when prosthetic devices cannot be used. SUMMARY: In the last few years, surgery of congenital mitral valve lesions has gained from echocardiography, which shows the exact function and anatomy of the mitral valve. The tendency is to avoid multistage operations. Valve replacement by biologic material (Ross II) is still under clinical evaluation. PMID- 16470144 TI - Heart transplantation for end-stage valvular disease: indications and results. AB - PURPOSE OF REVIEW: The aim of this study is to appraise the indications of a small group of heart transplanted patients with valvular disease, to analyse both their particular issues and results compared with the etiologies of other transplanted patients. RECENT FINDINGS: Analysis of recent data shows that valvular patients represent between 3 and 5% of transplantation indications. This proportion of valvular patients had a tendency to decrease in many countries. These patients on the whole have undergone multiple reoperations. Pulmonary resistance analysis has to be especially rigorous for this group. A primary excess mortality is directly related to multiple reoperations. Mean and long-term results are then strictly comparable with other etiologies. There are a few very specific indications for recurrent endocarditis in this group. SUMMARY: Valvular patients represent a subgroup of transplanted patients with a slight primary excess mortality but with identical long-term results. PMID- 16470145 TI - The real impact of randomized clinical trials in heart valve surgery. AB - PURPOSE OF REVIEW: The aim of this article is to evaluate the findings of clinical trials in cardiac valve surgery and to determine the real impact in standard of care. Also, publications on randomized clinical trials were reviewed as to integrity and validity. Nineteen randomized clinical trials were identified in 11 areas of operative and clinical management. RECENT DEVELOPMENTS: The Veterans Affairs and Edinburgh Heart Valve Trials confirmed the guidelines for indications for bioprostheses and mechanical prostheses. Even current prostheses have advanced technologies but the same valve-related complications determine indications. Randomized clinical trials of mechanical prostheses failed to determine prosthesis superiority. Bioprostheses of specific manufacturers contribute sub-optimal hemodynamics in small sizes. Two trials showed lack of superiority between aortic stented and stentless bioprostheses. Autografts, not allografts, are indicated for children because of structural valve deterioration of allografts. Atrial ablation surgery with concomitant mitral valve reconstruction/replacement is safe and efficacious with at least two energy sources. Minimally invasive aortic valve replacement does not provide superior results to conventional surgery. Patient-managed anticoagulation provides the most favourable thromboembolic and hemorrhagic rates with mechanical prostheses. Prosthesis sewing cuff impregnation with a bactericidal agent to reduce the incidence of prosthetic valve endocarditis was stopped because of increased incidence of major paravalvular leak requiring reoperation. SUMMARY: Randomized clinical trials, although limited in number, have provided advancement of the standard of care. Randomized clinical trials are indicated in the management of mild to moderate ischemic mitral regurgitation and evaluation of new transcatheter technologies to conventional surgery. PMID- 16470146 TI - Combined percutaneous coronary intervention and valve surgery. AB - PURPOSE OF REVIEW: This article reviews unique advantages emerging in valvular heart disease as the technology of invasive cardiology and cardiac surgery begin to merge. RECENT FINDINGS: Minimally invasive valve surgery is increasing in popularity and has helped to reduce morbidity. In addition, preoperative or intraoperative treatment of coronary artery disease by a percutaneous approach has simplified operations and allowed more liberal use of non-traditional incisions. SUMMARY: Percutaneous intervention with drug-eluting stents has provided early evidence for decreased restenosis and improved long-term patency rates. At the same time, cardiac surgery has moved toward less invasive approaches performed in new imaging arenas known as 'hybrid' operating rooms. Combining these technological advances is providing unique solutions to valvular heart disease also requiring revascularization, and will likely become the next horizon for strategies in cardiovascular medicine. PMID- 16470147 TI - Cardiac retransplantation: an ethical dilemma. AB - PURPOSE OF REVIEW: The evolution of scientific advancements that paved the way for clinical cardiac transplantation spans the era of the 20 century, heart transplantation has revolutionized therapy for end-stage heart failure. Demand far exceeds supply, resulting in a long waiting period, and an increasing number of deaths while on a waiting list. The shortage of donors poses dilemmas for allocation of organs and managing the waiting list. RECENT FINDINGS: The disparity between the demand and supply for donor hearts makes cardiac retransplantation an ethical issue with some patients being allowed a second transplant while some patients are dying on the waiting list before receiving their first transplant, especially with overall sub-optimal outcomes compared with primary transplantation. SUMMARY: The cardiac transplant community is mandated to closely monitor the results of cardiac retransplantation to identify the appropriate candidate who should receive a retransplantation. PMID- 16470148 TI - Mechanical circulatory support therapy as a bridge to transplant or recovery (new advances). AB - PURPOSE OF REVIEW: Large pulsatile left ventricular assist devices have been used as bridge-to-transplant therapy for the past 20 years. Over the past 2 years, a number of smaller rotary pumps have been introduced into clinical trials in the United States, Europe, and Australia. These devices offer the potential for smaller operations, greater resistance to infection, and new opportunities for bridge-to-recovery therapy. RECENT FINDINGS: Ongoing trials with axial flow devices support greater durability and less device-related infection than the HeartMate XVE (Thoratec Corporation, Pleasanton, California, USA). A greater tendency for pump thrombus and a higher anticoagulation requirement, however, are disadvantages compared with the HeartMate device. SUMMARY: Large pulsatile left ventricular assist devices have been the mainstay of mechanical support. The combination of durability and smaller pump size has become a focus of mechanical circulatory support in the current era. Ongoing clinical trials in the United States with three rotary pumps in both bridge-to-transplant and destination trials will likely result in a major increase in options for circulatory support. At least five other small, durable devices have recently entered non-US trials and are poised for clinical studies in the US. The National Institutes of Health sponsored Interagency Registry for Mechanically Assisted Circulatory Support (INTERMACS) will facilitate the introduction of new technology and study patient and device outcomes. PMID- 16470149 TI - Should we be doing routine biopsy after heart transplantation in a new era of anti-rejection? AB - PURPOSE OF REVIEW: The endomyocardial biopsy has defined the diagnosis of rejection in cardiac transplantation and has historically been a vital tool when rejection rates following transplantation were high. Surveillance biopsies have been the cornerstone of post-transplant management, as signs or symptoms of rejection are non-specific. With significant improvements in immunosuppressive therapy, however, the incidence of clinically significant rejection has declined, bringing into question the need for routine surveillance biopsy. This article reviews the current role of the endomyocardial biopsy in the management of patients following cardiac transplantation. RECENT FINDINGS: The endomyocardial biopsy is also limited by sub-optimal interobserver reproducibility, a lack of consensus with regard to treating certain grades of rejection, and often a lack of histological findings in patients with hemodynamic compromise, which frequently responds to anti-rejection therapy. Recent refinements, however, have allowed improved diagnosis of antibody mediated rejection, a relatively recently recognized entity. Moreover, a number of non-invasive modalities have been investigated recently as potential substitutes for the endomyocardial biopsy in detecting rejection. SUMMARY: Despite the development of a variety of non invasive methods for the detection of rejection, the endomyocardial biopsy will remain important in the management of patients following cardiac transplantation, as non-invasive techniques are associated with low specificity for the diagnosis of rejection. A new standardized classification will likely improve the utility of the biopsy by simplifying interpretation of cellular rejection and importantly allowing recognition of antibody-mediated rejection. PMID- 16470150 TI - Cardiac allograft vasculopathy: pathology, prevention and treatment. AB - PURPOSE OF REVIEW: Cardiac transplantation is a recognized therapy for end-stage heart failure. Graft coronary artery disease is a chief determinant of long-term survival following cardiac transplantation. There are multiple purported etiologies for graft coronary artery disease including both immunologic and nonimmunologic factors. Immunologic factors include human leukocyte antigen mismatching, cytokine production, and activation of the cellular immune system. Nonimmunologic factors include diabetes, hypertension, hyperlipidemia, and cytomegalovirus infection, just to name a few. There are also donor and recipient factors including age, prior coronary artery disease in the donor heart, and mode of donor brain death. RECENT FINDINGS: The diagnosis of graft coronary artery disease is especially difficult, partially due to the de-innervated allograft, as well as to its inherent predilection to affect the medium-sized and smaller arteries in a concentric and diffuse nature. Conventional angiography can overlook this condition because of the lack of eccentric plaques in larger epicardial arteries. Intravascular ultrasonography, by contrast, is more sensitive in detecting graft coronary artery disease but is unable to visualize the entire arterial system. Treatment is challenging and often unrewarding, leading to re-transplantation. Prevention is therefore ideal and involves protection against endothelial injury before and during transplantation as well as after transplantation, with decreased ischemic time, aggressive attention to early rejection, risk factor modification, and close follow-up. SUMMARY: This review will look at the pathophysiology of graft coronary artery disease, current diagnostic and therapeutic choices, as well as existing and future directions. PMID- 16470151 TI - Xenotransplantation: an ethical dilemma. AB - PURPOSE OF REVIEW: Xenotransplantation is an attractive alternative to allogeneic transplantation because of the intractable shortage of donor organs. The latest published developments in this area are discussed as well as recent publications regarding the continuing debates on the ethics of xenotransplantation and the safeguards that should be imposed to assure the minimization of risk to the patient and to human societies. RECENT FINDINGS: Several laboratories are concentrating on the development of genetically manipulated donor pigs in order to bypass the immunological barriers against transplantation of porcine organs and cells into humans. Preliminary experiments with these new pig strains suggest that this strategy has a high likelihood of success with respect to hyperacute rejection. Ethical considerations have been extended to the establishment of regulatory, legal, and ethical frameworks that will be necessary to minimize the risk of xenotransplantation to the recipients, their families, and to the worldwide community. SUMMARY: Xenotransplantation, because of the complexity of the medical, ethical and legal issues, will likely remain a controversial issue. If the scientific problems are solved, the decision to proceed with clinical application of this technique will depend on a collective decision guided by ethical, regulatory, and legal frameworks established by consensus. PMID- 16470152 TI - Bibliography. Current world literature. Arrhythmias. PMID- 16470153 TI - Bibliography. Current world literature. Orthopedics. PMID- 16470154 TI - Thrombosis in children with malignancy. AB - PURPOSE OF REVIEW: The association between thrombosis and malignancy in adults is well known. Children are now surviving malignancies that previously resulted in mortality. Complications, however, occur including thrombosis that result in mortality and morbidity. This review will explore the association in children and discuss the epidemiology and evidence-based diagnosis and treatment of thrombosis in this cohort. RECENT FINDINGS: Thrombin is an important link between malignancy, metastases, and inflammation. The accurate diagnosis of thrombosis in the upper venous system is directly related to the diagnostic technique used. Antithrombotic treatment is challenging owing to higher risks of bleeding, for example, decreased platelet counts, tiffelitis, large vascular tumor, etc. SUMMARY: Thrombosis and malignancy are associated in children. Properly designed studies are urgently required to further define the epidemiology of thrombosis in different malignancies and to find the best way to diagnose and treat thrombosis in children. PMID- 16470155 TI - Early integration of pediatric palliative care: for some children, palliative care starts at diagnosis. AB - PURPOSE OF REVIEW: Pediatric palliative care, with its emphasis on symptom management and quality of life, is an important aspect of care of children with life-threatening illnesses. We review recent publications with implications for care of these children. RECENT FINDINGS: Invasive and life-sustaining measures continue to be part of care for many children with life-threatening illnesses, even at the end of life. While these measures may seem reasonable when recovery is possible, they may not fit with a family's preferences for end-of-life care. One possible cause of the prevalence of invasive measures in children at the end of life is that complex illness trajectories in children make it difficult to predict the timing of death. Inadequate communication by clinicians can also lead to poor preparation for the end-of-life period. Early integration of palliative care allows for improved symptom management, parental adjustment, and preparation for the end-of-life care period. Families who have the opportunity to prepare for the end-of-life period, including learning what to expect, are more likely to feel that their care has been of high quality. Bereaved parents also recognize the value of talking about death with their children. SUMMARY: Early integration of palliative care can allow children and families to make decisions about care that fit with their values, and should become a standard of care for all children with life-threatening illnesses. PMID- 16470156 TI - Advances in understanding the genetic basis for bone-marrow failure. AB - PURPOSE OF REVIEW: Inherited marrow failure syndromes (IMFSs) are rare genetic diseases with varying degrees of cytopenia. Many of the syndromes are also characterized by nonhematological manifestations and a high risk of cancer. This review summarizes recent advances in understanding the genetic background of the common IMFSs. RECENT FINDINGS: Over recent years, numerous known and novel genes have been found to be associated with IMFSs. Although the functions of the proteins are largely unknown, they are postulated to play critical roles in fundamental cellular processes such as DNA repair, telomere maintenance, RNA metabolism, ribosomal biogenesis, growth-factor-signaling pathways and cell survival. For example, the telomere-related genes, DKC1 and TERC, have been identified as causes of dyskeratosis congenita. Also, homozygosity for the common cancer-associated gene, BRCA2, has been found to cause a rare subtype of Fanconi anemia. SUMMARY: The knowledge of the genetics of IMFSs has started to be translated into clinical practice. The identification of IMFS-related genes provided new diagnostic tools and better classification of the various disorders. Also, these advances enabled the design of clinical trials using gene therapy and preimplantation genetic diagnosis followed by in-vitro fertilization for selection of suitable embryos for hematopoietic stem-cell transplantation. PMID- 16470157 TI - The Ponseti method for treatment of congenital club foot. AB - PURPOSE OF REVIEW: This review of the Ponseti technique for the treatment of congenital club foot covers a topic of recently renewed interest. Pediatric orthopedists and parents have become increasingly enthusiastic about the success of this technique, which has been practiced continuously at the University of Iowa since 1948 but only recently become widely utilized. RECENT FINDINGS: Current literature has emphasized the reproducibility of these excellent results among multiple centers throughout the world. In addition, the role of the Internet, and accessibility of medical information have been central elements in the development of this phenomenon. SUMMARY: The current research should convince physicians that all children should be managed by the Ponseti technique at the outset. This research should also reassure physicians and parents that the overwhelming majority of children with club feet can be successfully managed, without the need for major reconstructive surgery. PMID- 16470158 TI - Congenital muscular torticollis: current concepts and review of treatment. AB - PURPOSE OF REVIEW: The purpose of this review is to better understand the spectrum of disease in torticollis, which is the third most common pediatric orthopaedic diagnosis in childhood. Besides the benign muscular tightness of the sternocleidomastoid muscle leading to the classic head position, the differential diagnosis of the wry neck include sequelae to inflammatory, ocular, neurologic or orthopedic diseases. Patients present with a stiff and tilted neck, and therefore require a thorough and systematic work-up, including a complete physical and neurologic examination and cervical spine radiographs. RECENT FINDINGS: Recent findings show that magnetic resonance imaging of the brain and neck is no longer considered cost-effective, or necessary, in congenital muscular torticollis. Observation and physical therapy, with or without bracing, is usually an effective treatment in most cases, especially if instituted within the first year of life. Botox has recently been shown to be an effective intermediate method of treatment for more resistant cases of congenital muscular torticollis. In those presenting after the age of 1 year, there is an increased rate of sternocleidomastoid muscle lengthening. The lengthening may improve the range of motion, but not necessarily the plagiocephaly, facial asymmetry, or cranial molding. SUMMARY: It is important to differentiate muscular from nonmuscular torticollis. Congenital muscular torticollis is benign; missing a case of nonmuscular torticollis could be potentially life threatening. PMID- 16470159 TI - Pediatric tibia fractures: current concepts. AB - PURPOSE OF REVIEW: Fracture of the tibia is a common occurrence in children. The operative treatment of pediatric tibia fractures has undergone a recent change. However, there is no clear consensus regarding the superiority of one treatment option. RECENT FINDINGS: The literature clearly supports the fact that the vast majority of pediatric tibia fractures can and should be managed nonoperatively. This is secondary to their inherent stability. A variety of factors including fracture type, location, severity and patient age determine the best treatment options for a particular fracture. A thorough understanding of these factors and how they affect outcome, help the clinician formulate the proper plan of treatment. SUMMARY: A randomized prospective controlled trial will be necessary to establish which surgical options are superior for which type of pediatric tibia fracture. Until then, recent studies have indicated that flexible intramedullary nails may lead to a shorter time to union and a decreased rate of refracture when compared with external fixation of unstable tibial shaft fractures. What remains unclear are the specific indications and contraindication for the use of flexible nails. External fixation still remains a successful treatment option for unstable tibial shaft fractures. PMID- 16470160 TI - The management of osteoid osteoma: updates and controversies. AB - PURPOSE OF REVIEW: Osteoid osteoma is a benign painful bone tumor usually found in the lower extremities of children and young adults. It has been traditionally treated by surgical excision. Despite the small size of the lesion, the operative procedure for its removal can be extensive, but still sometimes remains incomplete. The purpose of this review is to highlight and discuss current developments in the management of osteoid osteoma. RECENT FINDINGS: During the past decade, efforts were deployed to minimize bone removal, lessen the risk of pathologic fracture and the need for bone grafting, and thereby shorten the period of convalescence. Improved methods for the precise localization of an osteoid osteoma with use of radioisotope scanning or computed tomography scan have made it possible to treat this lesion with more limited and effective operations, mainly in deep and non-easily accessible osteoid osteomas. SUMMARY: Although they bear the criticism of lacking histological proof for diagnosis of osteoid osteoma, minimally invasive techniques, such as computed tomography guided percutaneous radiofrequency thermal ablation and laser photocoagulation have become the methods of choice for the treatment of all localizations except those in contact with neural structures (awaiting further research and experience), provided that the diagnosis is based on a typical clinical, scintigraphic and computed tomography presentation. PMID- 16470161 TI - The widening scope of coronaviruses. AB - PURPOSE OF REVIEW: In the past 2 years, at least three distinct human coronaviruses have been discovered, including the etiological agent associated with severe acquired respiratory syndrome (SARS). These recently discovered viruses, with the exception of the SARS associated coronavirus (SARS-CoV), are likely to be common respiratory viruses and may be responsible for a substantial proportion of respiratory tract disease. RECENT FINDINGS: The SARS-CoV first appeared in 2002 and spread rapidly around the globe. Although the worldwide spread of SARS-CoV may have been halted, the emergence of this new virus demonstrates the potential threat represented by species-to-species transmission of coronaviruses. NL63, initially isolated from a young child with lower respiratory tract disease, represents a group of newly described group I coronaviruses that have been identified worldwide, which are associated with both upper and lower respiratory tract disease, particularly in young children. The distribution of HKU1, a newly identified group II coronavirus, is not yet established. NL63 and HKU1 are related to the common human coronaviruses 229E and OC43, respectively. SUMMARY: The discovery of at least three new human coronaviruses represents significant advances in the investigation of human respiratory tract disease. Further studies are required to fully define the impact of these new pathogens. PMID- 16470162 TI - Role of breast milk in acquisition of cytomegalovirus infection: recent advances. AB - PURPOSE OF REVIEW: Congenital infection with cytomegalovirus is a major cause of disability in newborns. Recently, there has been increased emphasis on the study of postnatally acquired cytomegalovirus infection. One route by which cytomegalovirus infections are acquired in newborns is via consumption of breast milk from cytomegalovirus-seropositive, lactating mothers. The purpose of this review is to summarize recent studies of breast-milk-acquired cytomegalovirus infections in newborns, particularly in low-birth-weight premature infants. RECENT FINDINGS: Nearly all cytomegalovirus-seropositive women will reactivate and shed cytomegalovirus during lactation, as demonstrated by sensitive polymerase chain reaction techniques, as well as by viral culture of breast milk. A substantial proportion of infants exposed to cytomegalovirus in breast milk will acquire a primary cytomegalovirus infection. Although acquisition of cytomegalovirus by this route is seldom of consequence in healthy term infants, cytomegalovirus infections in low-birth-weight premature infants have been demonstrated to cause symptomatic illness, including hepatitis, neutropenia, thrombocytopenia, and a 'sepsis-like' state. SUMMARY: Cytomegalovirus is commonly shed in human milk, and cytomegalovirus-seropositive women can transmit this infection via breast-feeding. The benefits of breast-feeding greatly outweigh the minimal risk, if any, of infections transmitted to term infants. Caution is warranted, however, in low-birth-weight premature infants, who are at increased risk of cytomegalovirus disease. Interventions to screen breast milk, or to attempt to render breast milk noninfectious through treatments such as freezing, may be warranted in high-risk premature infants. PMID- 16470163 TI - Fostering acceptance of human papillomavirus and herpes simplex virus vaccines among adolescents and parents. AB - PURPOSE OF REVIEW: Vaccines for human papillomavirus and herpes simplex virus are currently in development, and these vaccines could be an effective and cost effective strategy to control these diseases. Young adolescents are likely to be the target recipients of these vaccines. These vaccines, however, will only be successful in reducing the prevalence of these diseases if accepted. This manuscript will focus on the issues associated with the acceptability of these vaccines by healthcare providers, parents, and adolescents/young adults. RECENT FINDINGS: Recent studies with healthcare providers, parents, and adolescent/young adults have found support for the acceptability of the human papillomavirus and herpes simplex virus vaccines. Protecting adolescents from serious diseases may be a more important priority for parents than concern about how the disease is transmitted. SUMMARY: It is likely that vaccines for human papillomavirus and herpes simplex virus will find acceptance among these groups. Strategies to increase acceptability and foster access, however, will still be needed. PMID- 16470164 TI - Recent trends in meningococcal epidemiology and current vaccine recommendations. AB - PURPOSE OF REVIEW: In 2005, a new meningococcal vaccine, tetravalent meningococcal conjugate vaccine (MCV4) was approved for use. This review discusses the population for whom it is recommended and the logic underlying these recommendations. RECENT FINDINGS: The older meningococcal vaccine, composed of capsular polysaccharide, had limited use in civilian populations. The Centers for Disease Control and the American Academy of Pediatrics now recommend the routine vaccination of adolescents with MCV4. This recommendation is largely based on newer epidemiologic data showing a considerable risk of meningococcal disease in late adolescence, most of which is preventable with vaccine. CONCLUSION: The new MCV4 vaccine should be used in primary-care practices for the immunization of all adolescents and high-risk children and adults. In the future, further studies are likely to show it to be safe and effective for younger children too. PMID- 16470165 TI - The use of fluoroquinolones in children. AB - PURPOSE OF REVIEW: The fluoroquinolones are an important group of antibiotics widely used in the treatment of various infectious diseases in adults, as a result of an excellent spectrum of activity, good tissue penetration and convenient ways of administration. Their use in children is limited as a result of possible fluoroquinolone-induced joint/cartilage toxicity observed mainly in juvenile animal studies. RECENT FINDINGS: Fluoroquinolones were successfully used in immunocompromised children and also in those suffering from multidrug resistant Gram-negative infections (including neonatal infections and multidrug resistant enteric infections caused by Salmonella and Shigella spp.). The fluoroquinolones were shown to be efficacious and well tolerated in the treatment of complicated cases of acute otitis media. The emergence of Streptococcus pneumoniae with reduced susceptibility to fluoroquinolones has been described worldwide. No arthropathy associated with fluoroquinolone use in children was evident. SUMMARY: With the exception of cystic fibrosis and life-endangering infections, the use of fluoroquinolones in pediatrics should be limited to Gram negative neonatal meningitis, Salmonella and Shigella spp. infections, chronic suppurative otitis media and some cases of complicated acute otitis media. Unskilled use of fluoroquinolones in children, particularly in community-acquired lower respiratory infections, could accelerate the emergence of pneumococcal resistance. PMID- 16470166 TI - Office pediatrics: current perspectives on the outpatient evaluation and management of lower respiratory infections in children. AB - PURPOSE OF REVIEW: The outpatient evaluation and management of a child with lower respiratory infection (LRI) remain a challenge to clinicians worldwide. This update will discuss current problems and new developments in the outpatient evaluation and treatment of pediatric LRIs. RECENT FINDINGS: The cause of pediatric LRIs remains partially defined. Mixed infections and 'unknown' organisms may be important sources of clinical illness. A wider incidence of atypical bacteria LRIs (notably Mycoplasma pneumoniae) in children is now recognized. Viral LRIs from rhinoviruses and human metapneumovirus are increasingly detected in children. Human metapneumovirus may compound the clinical severity of pediatric LRIs, specifically in combination with respiratory syncytial virus. Innovations in testing for viral LRIs offer a promising tool for the outpatient evaluation and management of pediatric LRIs. SUMMARY: Neither clinical symptoms nor findings on chest radiographs can reliably distinguish children with bacterial LRIs from those with viral or atypical pathogens. The efficacy of outpatient antibiotic treatment of atypical bacteria LRIs remains unproven. Multiplex rapid viral testing may ultimately help to refine strategies for outpatient management. The outpatient treatment of viral LRIs remains limited to supportive care; the 'value' of bronchodilators, epinephrine, or corticosteroids for treatment of bronchiolitis is unconfirmed. PMID- 16470167 TI - Adolescent and adult pertussis: disease burden and prevention. AB - PURPOSE OF REVIEW: According to surveillance data provided by the Centers for Disease Control and Prevention, the rates of pertussis disease in adolescents and adults have been increasing. This is likely due to increased recognition and waning vaccine-induced immunity. RECENT FINDINGS: The presentation of pertussis in adolescents and adults is generally a persistent cough, but more serious complications have been reported. In addition, adolescents and adults often serve as sources of pertussis infection in infants and young children. SUMMARY: Acellular pertussis vaccines combined with diphtheria and tetanus toxoids have proven to be well tolerated, immunogenic and effective in reducing pertussis disease in adolescents and adults. These vaccines are currently being recommended to replace the booster diphtheria and tetanus toxoid vaccines in adolescents. Recommendations for the use of these vaccines in adults are still being formulated. PMID- 16470168 TI - Bibliography. Current world literature. Hematology and oncology. PMID- 16470169 TI - RhoC promotes human melanoma invasion in a PI3K/Akt-dependent pathway. AB - Overexpression of the small GTPase, RhoC, in various human cancers has been correlated with high metastatic ability and poor prognosis. Rho-kinase (ROCK) is an important effector of Rho GTPases. The oncogenic serine/threonine kinase Akt (also known as PKB) is a downstream effector of phosphatidylinositol-3 kinase (PI3K). Akt activation contributes to the neoplastic phenotype by promoting cell cycle progression, increasing antiapoptotic functions, and enhancing tumor cell invasion. Rho signaling via ROCK has been previously shown either to activate or to downregulate PI3K/Akt. Using a human radial growth phase melanoma cell line, WM35, we have established stable transfectants that overexpress RhoC (called WM35RhoC). We found that overexpression of RhoC increased phosphorylated-Akt (Ser473/474/472, pAkt) expression and promoted cell invasion. Inhibition of RhoC with C3 transferase downregulated pAkt expression and decreased cell invasion in these cells. In addition, inhibition of PI3K, Akt, or ROCK partially decreased invasion. Further, inhibition of PI3K but not ROCK decreased the pAkt level. These results suggest that RhoC promotes invasion in part via activation of a PI3K/Akt pathway, in a manner independent of ROCK signaling. We propose that RhoC promotes melanoma progression via separate mechanisms that regulate the PI3K/Akt pathway and the ROCK signaling pathway. PMID- 16470170 TI - Retinoid-induced epidermal hyperplasia is mediated by epidermal growth factor receptor activation via specific induction of its ligands heparin-binding EGF and amphiregulin in human skin in vivo. AB - Retinoids are widely used in the treatment of photoaging to stimulate dermal repair. However, retinoids also induce epidermal hyperplasia, which can lead to excessive scaling. Scaling is the major deterrent to topical retinoid therapy. Keratinocyte growth is strongly stimulated via ligand activation of EGFR. We examined regulation of EGFR ligands by retinoids and the role of EGFR in retinoid induced hyperplasia in human skin in vivo. Topical treatment of human skin with all-trans retinoic acid (tRA) induces EGFR ligands heparin-binding (HB)-EGF and amphiregulin (AR), and reduces betacellulin mRNA levels. Laser capture microdissection-coupled real-time reverse transcription-PCR reveals that tRA increases HB-EGF mRNA throughout the epidermis, whereas AR induction is limited to basal keratinocytes. Topical tRA activates extracellular signal-regulated kinase 1/2 (Erk1/2) downstream EGFR effectors in human skin in vivo. tRA increases the soluble forms of AR and HB-EGF proteins, and induces epidermal hyperplasia, in human skin organ culture. Neutralization of HB-EGF or AR with specific antibodies strongly reduces tRA-induced epidermal hyperplasia. Finally, inhibition of EGFR activation by genistein reduces epidermal hyperplasia caused by topical retinoid treatment. These data demonstrate the central role of EGFR activation in retinoid-induced epidermal hyperplasia, and suggest that EGFR inhibitors can mitigate retinoid-induced scaling. PMID- 16470171 TI - Epitopes in the linker subdomain region of envoplakin recognized by autoantibodies in paraneoplastic pemphigus patients. AB - Sera from paraneoplastic pemphigus (PNP) immunoprecipitate multiple antigens from human epidermal protein extract. In this study, we further characterized the autoantibodies in 12 PNP sera. Immunoblotting using recombinant linker subdomains of envoplakin, periplakin, desmoplakin, and bullous pemphigoid antigen I found that 11 of the 12 sera recognized linker subdomains of envoplakin and periplakin. We then synthesized 12 peptides covering the linker subdomain of envoplakin for ELISA. One of the peptides, peptide no. 8, was recognized by nine out of the 12 sera with a higher affinity. A method of ligand-receptor binding assay was designed and performed using this peptide labeled with fluorescence as the ligand. Peptide no. 8 bound to CD20+ cells in Castleman's tumors from the patients whose sera were positive to this peptide by ELISA. Our data suggest that the linker subdomain of plakin proteins may be one of the major areas recognized by PNP autoantibodies, and epitopes in the linker subdomain of envoplakin recognized by PNP autoantibodies with a high affinity are dispersed in several areas and are variable among PNP patients. We also demonstrate that B-lymphocyte clones specifically reacting to epidermal proteins exist in Castleman's tumors from PNP. PMID- 16470172 TI - Cutaneous microdialysis as a novel means of continuously stimulating eccrine sweat glands in vivo. AB - Previous studies of the pharmacological regulation of sweat gland function in humans have administered agonists or antagonists systemically, by local intradermal injection or by iontophoresis. This has not allowed prolonged or steady-state activation of sweat glands to be examined. In this study, we used the technique of dermal microdialysis to administer pharmacological agents singly and in combination for up to 5 hours. Muscarinic stimulation with pilocarpine nitrate (50 mug ml(-1) to 1.66 mg ml(-1)) produced a sigmoid dose response curve, with maximal sweating (measured as transepidermal water loss) (mean 70 g m(-2) hour(-1)) after 15 minutes. This was sustained at steady-state levels (55 g m(-2) hour(-1)) until perfusion stopped. Perfusion with atropine (0.003 mg ml(-1)) reduced sweating below baseline and blocked pilocarpine-induced sweating completely. Noradrenaline (0.005 mg ml(-1)) induced much lower sweat rates than pilocarpine (56.8+/-1.62 g m(-2) hour(-1) vs 8.2+/-1.2 g m(-2) hour(-1), respectively, P<0.001) and this was unaffected by co-administration of atropine. This method has made it possible to show that sweat glands are capable of sustaining near maximal activity for at least 5 hours. The method has future application in investigation of conditions with disordered sweat gland activity. PMID- 16470173 TI - Clonal persistence and evolution during a decade of recurrent melanoma. AB - A patient with metastatic cutaneous melanoma responsive to immunotherapy experienced several recurrences over a decade of observation. With each recurrence, biopsies were obtained and cell lines generated. A rare mutation of the beta-catenin gene and an unbalanced methylation of the androgen receptor were documented in all cell lines. Karyotyping and comparative genomic hybridization identified consistent genetic traits in spite of divergent phenotypes, suggesting that all the metastases were derived from the same primary tumor, although they were each probably not derived from the most recent previous metastasis in a sequential manner. Thus, metastatic melanoma recurs from a common progenitor cell and phenotypic changes occur around a central core of genetic stability. This observation may bear significance for the development of targeted anticancer therapies. PMID- 16470174 TI - In vivo and ex vivo UV-induced analysis of pigmentation gene expressions. PMID- 16470175 TI - Heavy water labeling of keratin as a non-invasive biomarker of skin turnover in vivo in rodents and humans. AB - Measurement of skin turnover has been problematic in humans. Heavy water (2H2O) labeling has recently been developed as a safe, simple method to study in vivo kinetics of many biosynthetic processes, including DNA and protein synthesis. Here, we apply this approach to the measurement of 2H incorporation into skin keratin and show close agreement between keratin and keratinocyte turnover data in the epidermis of rodents. Elevated turnover rates of both keratin and keratinocytes were observed in the epidermis of the flaky skin mouse, although topical treatments effective in human psoriasis had no effect on either turnover rate in these mice. In humans, keratin turnover was monitored non-invasively by serial tape stripping during and after 2H2O labeling. Kinetic data were consistent with previous estimates of epidermal turnover, with a lag time of 18 days before label appeared at the skin surface and a transit time of 4-5 weeks. Variability in skin keratin turnover rates was present among healthy individuals. In summary, 2H2O labeling of skin keratin represents a non-invasive approach for assessing skin turnover dynamics in pre-clinical models and in human subjects. PMID- 16470176 TI - FK506 controls CD40L-induced systemic autoimmunity in mice. AB - Autoimmunity results from loss of mechanisms controlling self-reactivity. Autoimmune disorders play an increasingly important role and CD40-CD40 ligand (CD40L) interaction on immunocompentent cells is able to break established immunotolerance. To study the effects of the calcineurin-inhibitor FK506 on CD40L induced systemic autoimmunity, CD40L transgenic (tg) mice, which spontaneously develop a mixed connective tissue-like disease, were treated with FK506 after onset of overt autoimmunity. Interestingly, FK506-treated CD40L tg mice showed significantly reduced autoimmune dermatitis scores and markedly decreased numbers of lesional infiltrating leukocytes. This finding was associated with diminished lymphadenopathy induced by FK506 treatment. Furthermore, FK506 suppressed the development of cytotoxic/autoreactive CD8(+) T cells as evidenced by the reduced expression of T cell activation markers in treated CD40L tg mice. Importantly, FK506 induced a significant reduction in autoantibody titers in the serum of CD40L tg animals. As CD40L tg mice develop nephritis leading to loss of renal function proteinuria was determined after FK506 injections. Notably, FK506 treatment re-established renal function as indicated by significantly reduced uric protein concentrations of treated CD40L tg mice. Together, these findings show the beneficial therapeutic effects of FK506 for the treatment of CD40L induced autoimmunity. Additionally, these results demonstrate that FK506 is able to suppress ongoing severe autoimmune responses. PMID- 16470177 TI - Increased JunB mRNA and protein expression in psoriasis vulgaris lesions. PMID- 16470178 TI - Delayed cutaneous wound healing in mice lacking solute carrier 11a1 (formerly Nramp1): correlation with decreased expression of secretory leukocyte protease inhibitor. AB - Control of macrophage functions by natural resistance-associated macrophage protein 1 (NRAMP1) has proven to be important for murine resistance to several intracellular pathogens, including Mycobacterium bovis BCG and Salmonella typhimurium, although the exact molecular mechanism of its action remains unknown. We identified secretory leukocyte protease inhibitor (SLPI) as a novel candidate gene whose expression is dependent on Nramp1 gene expression using B10A.Nramp1+/+ and B10A.Nramp1-/- macrophage cell lines in vitro, as well as mice bearing the resistance alleles (wild type (WT)) of the Nramp1 and mice with an ablated Nramp1 gene (knockout (KO)). We established that B10A.Nramp1+/+ cells spontaneously expressed a 10-fold higher level of SLPI messenger RNA (mRNA) compared to B10A.Nramp1-/- expression. Similarly, protein secretion was detected only in supernatants from B10A.Nramp1+/+ macrophages. Induction of SLPI in excisional cutaneous wounds and, most importantly, in macrophages infiltrating these wounds was significantly higher in Nramp1 WT mice compared to KO mice. These differences in SLPI expression in vivo correlated with a significant delay in the kinetics of wound healing in Nramp1 KO mice compared to WT controls. Taken together, these results suggest for the first time that Nramp1 controls macrophage SLPI mRNA and protein expression, and can also have an important effect on the kinetics of wound healing. PMID- 16470179 TI - Doxorubicin-induced alopecia is associated with sebaceous gland degeneration. AB - Alopecia, accompanied by skin dryness, is one of the distressing side effects often occurring in chemotherapy-treated cancer patients. Little is known of the effects of chemotherapy on sebaceous glands, despite their importance in hair follicle homeostasis. This study investigates sebaceous gland morphology and the response of SZ95 sebaceous gland cell line to doxorubicin (DXR) treatment. The morphology of sebaceous glands during intraperitoneal DXR treatment was investigated by optical and electron microscopy in a 7-day-old rat model and further confirmed in an adult mouse model. Moreover, in vitro studies using the SZ95 sebaceous gland cell line were performed to assess the response of sebocytes to DXR in terms of cell proliferation, apoptosis, and necrosis. DXR treatment induced sebaceous gland regression and occasionally caused their complete disappearance. This observed damage and disappearance preceded DXR-induced hair loss. In vitro experiments using the SZ95 sebaceous gland cell line indicated that DXR treatment induced a differentiation process leading to premature sebocytes apoptosis. Owing to the importance of the sebaceous gland in hair follicle homeostasis, DXR-induced involution of this gland might be related to subsequent hair loss. PMID- 16470180 TI - Functional characterization and expression analysis of the proteinase-activated receptor-2 in human cutaneous mast cells. AB - Proteinase-activated receptor-2 (PAR2) belongs to a new G protein-coupled receptor subfamily activated by serine proteinases. PAR2 has been demonstrated to play a role during inflammation and immune response in different tissues including the skin. We examined whether PAR2 is functionally expressed by cutaneous human primary skin mast cells (HPMC) and the human mast cell line 1 (HMC-1). Reverse transcription-polymerase chain reaction and FACS analysis show expression of PAR2 both at the RNA and protein level. HPMCs and HMC-1 also express PAR1, PAR3, and PAR4. Ca-mobilization studies demonstrate functional PAR2 expressed by human skin mast cells, as shown by natural and synthetic PAR2 agonists. PAR2 agonists induced histamine release from HPMC indicating a role of PAR2 in regulating inflammatory and immune responses by skin mast cells. Double immunofluorescence staining reveals colocalization of PAR2 with tryptase in the majority of human skin mast cells. In conclusion, trypsin and tryptase as well as specific agonists for PAR2 were able to induce Ca2+ mobilization in HPMCs, and agonists of PAR2 induce the release of histamine from these cells. Thus, PAR2 may be an important regulator of skin mast cell function during cutaneous inflammation and hypersensitivity. PMID- 16470181 TI - Circulating tumor cells and detection of the melanoma-associated antigen HMW-MAA in the serum of melanoma patients. PMID- 16470182 TI - Genomic analysis defines a cancer-specific gene expression signature for human squamous cell carcinoma and distinguishes malignant hyperproliferation from benign hyperplasia. AB - Using high-density oligonucleotide arrays, we measured expression of >12,000 genes in surgical excisions of invasive human squamous cell carcinomas (SCCs) versus site-matched control skin. This analysis defined >1,900 genes with altered expression in SCCs that were statistically different from controls. As SCCs are composed of epithelial cells, which are both hyperplastic and invasive, we sought to define gene sets associated with these biologic processes by comparing gene expression to psoriasis vulgaris, which is a condition of benign keratinocyte hyperplasia without invasiveness or pre-malignant potential. Through this analysis, we found genes that were commonly upregulated in both conditions and unique genes with increased expression in SCCs. Differential gene regulation in these two conditions was confirmed by real-time reverse transcription-PCR and immunohistochemistry. We found that benign hyperplasia is associated with upregulation of genes including DEFB4 (defensin B4), SERPINB3 (serine proteinase inhibitor, member 3), STAT1 (signal transducer and activator of transcription 1), K16 (keratin 16), CEACAMs (carcinoembryonic antigen-related cell adhesion molecules), and WNT 5A (wingless-type MMTV integration site family, member 5A). WNT receptor frizzled homolog 6 (FZD6) and prostaglandin-metabolizing enzyme hydroxyprostaglandin dehydrogenase were increased in SCC alone. Growth factor pleiotrophin (PTN) was expressed at higher levels in non-tumor-bearing skin adjacent to excised SCC. SCC was further characterized by upregulation of matrix metalloproteinases 1, 10, and 13, cathepsin L2, cystatin E/M as well as STAT3 and microseminoprotein, beta (MSMB), and downregulation of inducible nitric oxide synthase, granzyme B, CD8, and CD83. The current study defines a unique gene expression signature for cutaneous SCC in humans and suggests potential roles for WNT, FZD, and PTN in the pathogenesis of SCC. PMID- 16470184 TI - Supporting the support groups. PMID- 16470183 TI - An important role of lymphatic vessels in the control of UVB-induced edema formation and inflammation. PMID- 16470185 TI - Prevention of retrograde stone migration during ureteroscopy. PMID- 16470186 TI - Transobturator versus retropubic suburethral tapes for stress urinary incontinence. PMID- 16470201 TI - Can an erectogenic pharmacotherapy regimen after radical prostatectomy improve postoperative erectile function? PMID- 16470200 TI - Does postoperative adjuvant radiotherapy reduce biochemical progression after radical prostatectomy? PMID- 16470202 TI - What are the operating characteristics of PSA screening for prostate cancer? PMID- 16470203 TI - Extracorporeal shock-wave lithotriptors: why newer may not be better. PMID- 16470204 TI - Comparison of biochemical progression rates after radical prostatectomy and radiotherapy for localized prostate cancer. PMID- 16470205 TI - Is a combination of alarm and drug therapies effective in nocturnal enuresis? PMID- 16470206 TI - Does addition of 13-cis-retinoic acid to interferon-alpha2a improve survival in patients with metastatic RCC? PMID- 16470207 TI - How, why and when should urologists evaluate male sexual function? AB - Male sexual dysfunction-a term that is commonly used to refer to erectile dysfunction, premature ejaculation, decreased libido and impaired orgasm-is the primary complaint encountered by many urologists. Despite the high prevalence and bothersome nature of these complaints, they are frequently neglected in clinical practice. This paper highlights clinical situations in which urologists should systematically evaluate male sexual functioning. These include men who present with several common urologic disorders, such as pelvic trauma, malignancies, and lower urinary tract symptoms associated with benign prostatic hyperplasia, neurologic disorders and infertility. Studies have shown that erectile dysfunction might be a clinical marker of endothelial dysfunction, and consequently of undetected diabetes, hypertension, dyslipidemia, coronary artery disease and depression. We also address the question of whether urologists should adopt wide-ranging screening regimens for sexual dysfunction. PMID- 16470208 TI - Surgery Insight: advantages and pitfalls of surgical techniques for the correction of bladder exstrophy. AB - Staged reconstruction repair of bladder exstrophy results in hydronephrosis or renal scarring in 15-25% of patients. A cosmetically acceptable and functional phallus can be achieved in 85% of patients, 20-30% of whom will require more than one operation for penile reconstruction. Episodes of penile glans loss or corporal loss are rarely reported with this technique. Widely disparate results relating to complete urinary continence and volitional voiding have been published, with urinary continence reported to occur in 7-85% of patients. The need for bladder augmentation to obtain urinary continence also varies, with reports that somewhere between 10% and 90% of patients require an augmentation procedure to gain urinary continence. Complete primary repair of bladder exstrophy using the penile disassembly technique results in hydronephrosis or renal scarring in 0-30% of patients, and hypospadias, as a consequence of this repair, will occur in 30-70% of patients. Loss of the glans and corpora appear more frequently with penile disassembly than in staged reconstruction of bladder exstrophy, however, the exact incidence of this complication is unknown. Reported complete urinary continence and volitional voiding rates are also varied following penile disassembly, ranging from 25-65%. A modified bladder-neck reconstruction to gain urinary continence is reportedly required in 15-90% of patients, with 5-10% requiring both bladder augmentation and bladder-neck reconstruction. Experience with complete primary repair of bladder exstrophy, using the penile disassembly approach, seems promising but is not a panacea. To outline the risks and benefits regarding the various surgical techniques for bladder exstrophy, we would recommend the establishment of a national registry for patients with this disorder. PMID- 16470209 TI - Mechanisms of Disease: the role of nerve growth factor in the pathophysiology of bladder disorders. AB - The case is compelling for the involvement of nerve growth factor (NGF) in the pathogenesis of lower urinary tract disease, especially in conditions with altered neural function. Remodeling of the micturition pathways occurs following experimental bladder-outlet obstruction, denervation, spinal cord injury, cystitis, and diabetes mellitus. Clinically, NGF levels are elevated in the bladders of men with benign prostatic hyperplasia, women with interstitial cystitis and in patients with idiopathic overactive bladder. Blockade of NGF, using either an endogenous antibody or an antibody against the NGF receptor, prevents neural plasticity and bladder overactivity in experimental models of these conditions. The ability of NGF to trigger bladder overactivity might rely on altering the properties of sodium or potassium channels (or their expression) in bladder afferent fibers. Therapies based on altered NGF levels, or changes in channel properties in afferent nerves, represent an intriguing avenue of investigation for the management of detrusor overactivity or diabetic cystopathy. PMID- 16470210 TI - Treatment of Peyronie's disease with oral pentoxifylline. AB - BACKGROUND: A 51-year-old male presented with a penile deformity without obvious etiology. Physical examination revealed hard plaques in the dorsal and ventral penis. INVESTIGATIONS: Physical examination, penile ultrasound. DIAGNOSIS: Peyronie's disease. MANAGEMENT: Oral pentoxifylline. PMID- 16470211 TI - Inducible nitric oxide synthase activity is essential for inhibition of prostatic tumor growth by interferon-beta gene therapy. AB - We have previously reported that adenoviral vector-mediated interferon (IFN)-beta gene therapy inhibits orthotopic growth of human prostate cancer cells in nude mice. The purpose of this study was to determine efficacy and mechanisms of this therapy in immune-competent mice. TRAMP-C2Re3 mouse prostate cancer cells infected with 100 multiplicity of infection (MOI) of adenoviral vector encoding for mouse IFN-beta (AdmIFN-beta), but not AdE/1 (a control adenoviral vector), produced approximately 60 ng/10(5) cells/24 h of IFN-beta. The tumorigenicity of AdmIFN-beta-transduced cells was dramatically reduced in the prostates of C57BL/6 mice. A single intratumoral injection of 2 x 10(9) PFU (plaque-forming unit) of AdmIFN-beta inhibited tumor growth by 70% and prolonged survival of tumor-bearing mice. Intriguingly, this AdmIFN-beta therapy did not alter the growth of tumors in inducible nitric oxide synthase (iNOS)-null C57BL/6 mice. Immunohistochemical analysis revealed that treatment of tumors with AdmIFN-beta in wild-type C57BL/6 mice led to increased iNOS expression, decreased microvessel density, decreased cell proliferation, and increased apoptosis. Furthermore, quantitative reverse transcriptional PCR analysis showed that AdmIFN-beta therapy, in C57BL/6 but not the iNOS-null counterparts, reduced levels of the mRNAs for angiopoietin, basic fibroblast growth factor, matrix metalloproteinase-9, transforming growth factor beta1, vascular endothelial growth factor (VEGF)-A, and VEGF-B, as well as the antiapoptotic molecule endothelin-1. These data indicated that IFN-beta gene therapy could be effective alternative for the treatment of locally advanced prostate cancer and suggest an obligatory role of NO in IFN-beta antitumoral effects in vivo. PMID- 16470214 TI - The first cut is the deepest: basic surgical training in ophthalmology. PMID- 16470212 TI - Modification of the p53 transgene of a replication-competent adenovirus prevents mdm2- and E1b-55kD-mediated degradation of p53. AB - Clinical efficacy of adenovirus-mediated cancer gene therapy has been limited thus far. To improve its oncolytic effect, a replication-competent adenoviral vector was previously constructed to express high levels of p53 at a late time point in the viral life cycle. p53 expression from this vector improved tumor cell killing and viral spread in vitro. However, p53 function is antagonized by cellular mdm2 and adenoviral E1b-55kD, both of which are known to bind to and inactivate p53. Therefore, a new vector (Adp53W23S) that expresses a modified p53 transgene, which does not bind to E1b-55kd and mdm2, was constructed. The modified p53 protein was demonstrated to have a substantially prolonged half life, and its localization was predominantly nuclear. Viral replication was unaffected by expression of the modified p53 and cancer cell killing was improved in vitro. However, in a xenograft model, efficacy was not significantly different from control virus. In conclusion, expression of a degradation-resistant p53 transgene late in the life cycle of a replication-competent adenovirus improves p53 stability and cancer cell killing in vitro. However, other factors, such as the adenoviral E1b-19kD and E1a proteins, which oppose p53 function, and limitations to viral spread need to be addressed to further improve in vivo efficacy. PMID- 16470215 TI - Acute subretinal macular haemorrhage following an accidental electrical shock. PMID- 16470216 TI - Accuracy of choroidal melanoma diagnosis by general ophthalmologists: a prospective study. AB - PURPOSE: To estimate the proportion of 'false positives' in patients referred with a diagnosis of suspected choroidal melanoma by general ophthalmologists to an ocular oncology centre. METHODS: A prospective study of patients referred by general ophthalmologists to an ocular oncology centre was undertaken over a 14 week period. The diagnosis was made clinically in patients receiving radiotherapy or phototherapy and was confirmed by histopathology in patients requiring fine needle aspiration biopsy (FNAB) or enucleation. RESULTS: A total of 132 new patients were seen in 10 consecutive ocular oncology clinics between 29 March 2004 and 5 July 2004. The mean age was 62 years (range 28-88 years) and 60 (55%) were female. Among the 83 suspected malignant posterior segment lesions, the suspected diagnosis included choroidal melanoma (73), choroidal metastasis (6), 'choroidal tumour' (3), and 'Solid retinal detachment' (1). Only 50 of the 73 suspected melanomas were confirmed (68.5%; 95% CI, 57-78%), the oncologist's diagnosis in the remaining 23 being choroidal naevus (10), choroidal metastasis (1), circumscribed choroidal haemangioma (2) and others (10). Only one of six patients with suspected metastases had this condition, the remainder having melanoma (1), lymphoma (1), circumscribed choroidal haemangioma (1), and others (2). The 'choroidal tumours' and 'solid detachments' were found to be chorio retinal disciform scar (1), varix of vortex vein (1), eccentric CNV (1), and subretinal haemorrhage (1) CONCLUSION: Approximately 30% of patients referred to an ocular oncology service with the diagnosis of choroidal melanoma have an incorrect diagnosis. PMID- 16470217 TI - An atypical presentation of Wernicke's encephalopathy in an 11-year-old child. PMID- 16470219 TI - Ocular surface squamous neoplasia in a renal transplant recipient on immunosuppressive therapy. PMID- 16470220 TI - Comment on 'smoking delays the response to treatment in episcleritis and scleritis'. PMID- 16470221 TI - Solitary nasal neurofibroma presenting as compressive optic neuropathy. PMID- 16470223 TI - Dissection of the NF-kappaB signalling cascade in transgenic and knockout mice. AB - Studies in transgenic and knockout mice have made a major contribution to our current understanding of the physiological functions of the NF-kappaB signalling cascade. The generation and analysis of mice with targeted modifications of individual components of the NF-kappaB pathway tremendously advanced our knowledge of the roles of the NF-kappaB proteins themselves, and also of the many activators and negative regulators of NF-kappaB. These studies have highlighted the complexity of the NF-kappaB system, by revealing the multiple interactions, redundancies, but also diverse functions, performed by the different molecules participating in the regulation of NF-kappaB signalling. Furthermore, inhibition or enforced activation of NF-kappaB in transgenic mice has uncovered the critical roles that NF-kappaB plays in the pathogenesis of various diseases such as liver failure, diabetes and cancer. PMID- 16470224 TI - Specific resistance upon lentiviral TRAIL transfer by intracellular retention of TRAIL receptors. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) selectively induces apoptosis in many transformed cells, suggesting TRAIL as an ideal candidate for cancer gene therapy. A main obstacle in cancer therapy is intrinsic or acquired therapy resistance of malignant cells. To study induction of resistance against TRAIL, we generated lentiviral vectors allowing efficient TRAIL expression and apoptosis induction in a variety of human cancer cell lines. Within days upon TRAIL overexpression, cells became resistant towards TRAIL, but not to CD95 ligation or DNA damage by cisplatin. Cell surface expression of TRAIL receptors 1 and 2 was completely abrogated in resistant cells due to intracellular retention of the receptors by TRAIL. SiRNA directed against TRAIL resensitized the resistant cells by restoring cell surface expression of TRAIL receptors. These findings represent a novel resistance mechanism towards TRAIL, specifically caused by TRAIL overexpression, and question the use of TRAIL expression in tumor-cell targeting gene therapy. PMID- 16470222 TI - RhoA GTPase inactivation by statins induces osteosarcoma cell apoptosis by inhibiting p42/p44-MAPKs-Bcl-2 signaling independently of BMP-2 and cell differentiation. AB - Osteosarcoma is the most common primary bone tumour in young adults. Despite improved prognosis, resistance to chemotherapy remains responsible for failure of osteosarcoma treatment. The identification of signals that promote apoptosis may provide clues to develop new therapeutic strategies for chemoresistant osteosarcoma. Here, we show that lipophilic statins (atorvastatin, simvastatin, cerivastatin) markedly induce caspases-dependent apoptosis in various human osteosarcoma cells, independently of bone morphogenetic protein (BMP)-2 signaling and cell differentiation. Although statins increased BMP-2 expression, the proapoptotic effect of statins was not prevented by the BMP antagonist noggin, and was abolished by mevalonate and geranylgeranylpyrophosphate, suggesting the involvement of defective protein geranylgeranylation. Consistently, lipophilic statins induced membrane RhoA relocalization to the cytosol and inhibited RhoA activity, which resulted in decreased phospho-p42/p44- mitogen-activated protein kinases (MAPKs) and Bcl-2 levels. Constitutively active RhoA rescued phospho p42/p44-MAPKs and Bcl-2 and abolished statin-induced apoptosis. Thus, lipophilic statins induce caspase-dependent osteosarcoma cell apoptosis by a RhoA-p42/p44 MAPKs-Bcl-2-mediated mechanism, independently of BMP-2 signaling and cell differentiation. PMID- 16470225 TI - Anti-idiotype-mediated epitope spreading and diminished phagocytosis by a human monoclonal antibody recognizing late-stage apoptotic cells. AB - Apoptotic cells are considered an important auto-antigenic source in diseases such as systemic lupus erythematosus (SLE). A human monoclonal antibody demonstrating exquisite specificity towards late-stage apoptotic cells was generated from an SLE patient. Polyreactive recognition of ribonucleoproteins Ro52 and Ro60 was observed. The antibody significantly diminished the phagocytosis of apoptotic cells and a concomitant decrease in transforming growth factor-beta (TGF-beta) secretion was observed. Light and heavy chain sequencing revealed the antibody to be in essentially germline configuration. Elicited anti idiotypic antibodies bound distinct self-antigens and showed augmented reactivity towards apoptotic cells as well. Thus, near-germline encoded antibodies recognizing antigens externalized during the process of apoptosis can mediate a variety of potentially pathogenic effects; decreases in the phagocytic uptake of dying cells would constitute a disease-perpetuating event and stimulation of the idiotypic network could lead to intermolecular epitope spreading, increasing the range of molecular targets.. PMID- 16470226 TI - Integrated signalling pathways for mast-cell activation. AB - Mast-cell activation mediated by the high-affinity receptor for IgE (FcepsilonRI) is considered to be a key event in the allergic inflammatory response. However, in a physiological setting, other receptors, such as KIT, might also markedly influence the release of mediators by mast cells. Recent studies have provided evidence that FcepsilonRI-dependent degranulation is regulated by two complementary signalling pathways, one of which activates phospholipase Cgamma and the other of which activates phosphatidylinositol 3-kinase, using specific transmembrane and cytosolic adaptor molecules. In this Review, we discuss the evidence for these interacting pathways and describe how the capacity of KIT, and other receptors, to influence FcepsilonRI-dependent mast-cell-mediator release might be a function of the relative abilities of these receptors to activate these alternative pathways. PMID- 16470227 TI - Type A and B RNase P RNAs are interchangeable in vivo despite substantial biophysical differences. AB - We show that structural type A and B bacterial ribonuclease P (RNase P) RNAs can fully replace each other in vivo despite the many reported differences in their biogenesis, biochemical/biophysical properties and enzyme function in vitro. Our findings suggest that many of the reported idiosyncrasies of type A and B enzymes either do not reflect the in vivo situation or are not crucial for RNase P function in vivo, at least under standard growth conditions. The discrimination of mature tRNA by RNase P, so far thought to prevent product inhibition of the enzyme in the presence of a large cellular excess of mature tRNA relative to the precursor form, is apparently not crucial for RNase P function in vivo. PMID- 16470228 TI - Inhibition of apoptosis by survivin improves transplantation of pancreatic islets for treatment of diabetes in mice. AB - Survivin is a cancer gene implicated in inhibition of apoptosis and regulation of mitosis, but its function in normal cells has remained elusive. Here, we show that transgenic mice expressing survivin in pancreatic islet beta-cells show no changes in cell proliferation, as determined by islet size or islet number. Transplantation of survivin transgenic islets in diabetic recipient mice affords long-term engraftment and stable correction of hyperglycaemia. This involves intrinsic inhibition of beta-cell apoptosis, in vivo, and global transcriptional changes in pancreatic islets with upregulation of stress response genes, antagonists of cytokine signalling and promoters of angiogenesis. These broad cytoprotective functions of survivin in vivo might be beneficial for gene therapy of diabetes. PMID- 16470230 TI - Impact of adjuvant breast cancer chemotherapy on fatigue, other symptoms, and quality of life. AB - PURPOSE/OBJECTIVES: To identify differences in fatigue, other physical symptoms, and psychological symptoms and their relationship to quality of life (QOL) during chemotherapy and as long as one year after. DESIGN: Longitudinal, descriptive design embedded in a pilot intervention study. SETTING: Midwestern urban oncology clinics and patient homes. SAMPLE: 25 Caucasian women, aged 40-65 years (-X = 54.3), with stage I or II breast cancer receiving doxorubicin-based chemotherapy. METHODS: The Piper Fatigue Scale, Hospital Anxiety and Depression Scale, Symptom Experience Scale, and Medical Outcomes Study Short-Form General Health Survey were completed before and after each treatment; 30, 60, and 90 days after the last treatment; and one year after the first treatment. MAIN RESEARCH VARIABLES: Fatigue, physical and psychological symptoms, and QOL. FINDINGS: Fatigue levels were moderately intense during treatments and decreased significantly over time. Sleep disturbances and pain were the most frequent, intense, and distressing other physical symptoms. Anxiety was highest at baseline, and depression was highest during the fourth chemotherapy treatment. Fatigue was correlated with other physical and psychological symptoms at some times during treatments and consistently following treatments. Higher fatigue was associated with lower QOL in several domains. CONCLUSIONS: Fatigue is associated with other physical and psychological symptoms that fluctuate during and after treatment. Higher fatigue compromises QOL. IMPLICATIONS FOR NURSING: Interventions targeting primary or cluster symptoms can reduce the impact of adjuvant chemotherapy on fatigue, other symptoms, and QOL. PMID- 16470229 TI - Effects of spirituality in breast cancer survivors. AB - PURPOSE/OBJECTIVES: To examine the effects of spirituality (meaning in life and prayer) on a sense of well-being among women who have had breast cancer. DESIGN: Descriptive, correlational, cross-sectional. SETTING: Rural and urban communities in central Texas. SAMPLE: Convenience sample of 84 women, 34-80 years of age diagnosed less than one year previously (36%), within the previous one to five years (38%), and more than five years previously (26%). METHODS: The study approach consisted of completing a questionnaire assessing personal and cancer characteristics, aspects of spirituality (meaning in life and prayer), and physical and psychological responses to breast cancer. MAIN RESEARCH VARIABLES: Meaning in life, prayer, and physical and psychological responses to breast cancer. FINDINGS: Meaning in life was positively related to psychological responses and negatively related to physical responses. Prayer was positively related to psychological well-being. Women with higher prayer scale scores reported lower education levels, less income to meet their needs, and closer relationships with God. Meaning in life mediated the impact of breast cancer on physical and psychological well-being. CONCLUSIONS: Strong relationships exist among spirituality and personal and cancer characteristics. Meaning in life mediated the effects of breast cancer on well-being in breast cancer survivors. IMPLICATIONS FOR NURSING: The findings support healthcare providers encouraging women diagnosed with breast cancer to explore their spirituality as an effective resource for dealing with the physical and psychological responses to cancer. PMID- 16470232 TI - Cancer nursing. PMID- 16470231 TI - Seeking and managing hope: patients' experiences using the Internet for cancer care. AB - PURPOSE/OBJECTIVES: To describe the experiences of patients with cancer using the Internet for information and support to manage the self-care aspects of illness and treatment, including symptom management. RESEARCH APPROACH: Heideggerian hermeneutics branch of phenomenology. SETTING: The interviews took place in outpatient settings in the northeastern United States, including clinics, patients' homes, and the researchers' office. PARTICIPANTS: 20 patients self identified as users of the Internet for cancer care. METHODOLOGIC APPROACH: Data were collected by informal interviews that provided the narrative stories for hermeneutic analysis. MAIN RESEARCH VARIABLES: Internet use for cancer care and patient-provider relationship. FINDINGS: Five related themes and one constitutive pattern described patients' experiences. The themes were retrieving and filtering Internet information according to personal situation by Internet-savvy people in patients' support networks, seeking hope from the newest treatment options while coping with fear in manageable "bytes," self-care for personal illness situations with meaningful information regarding symptom management, empowering patients as partners when Internet information served as a second opinion in decision making and validating treatment decisions, and Internet as providing peer support. The constitutive pattern was Internet use as assisting patients in discovering ways to live with cancer as a chronic illness instead of a death sentence. CONCLUSIONS: Patients with cancer are incorporating Internet use into their cancer care. They perceive changing provider-patient relationships when they participate in treatment decisions. INTERPRETATION: Computer-savvy patients and their personal support networks will avail themselves of Internet information, creating the need for new interaction patterns and relationships with providers. PMID- 16470233 TI - Conceptualizing and measuring physical functioning in cancer survivorship studies. AB - PURPOSE/OBJECTIVES: To propose a conceptual model to guide the design of intervention studies to improve physical functioning in cancer survivors. DATA SOURCES: Conceptualizations of physical functioning in people without cancer and exercise studies in breast cancer survivors. DATA SYNTHESIS: Most exercise studies measured only one of three possible dimensions of physical functioning, and mediators seldom were tested. CONCLUSIONS: Careful selection, naming, and measurement of physical functioning outcomes could maximize generation of new knowledge. More frequent testing of mediators could show how interventions affect physical functioning. IMPLICATIONS FOR NURSING: Nurse scientists designing interventions to increase exercise, reduce symptoms, or manage side effects of treatment in cancer survivors are likely to measure physical functioning outcomes. Consistent measurement, terminology, and reporting of physical functioning outcomes in these studies will facilitate communication among nurse scientists and hasten translation of knowledge into clinical practice. PMID- 16470234 TI - Predicting and modifying substance use in childhood cancer survivors: application of a conceptual model. AB - PURPOSE/OBJECTIVES: To identify factors that predict or modify substance use in childhood cancer survivors and to describe how a risk-counseling intervention reduced young survivors' substance use. DESIGN: Secondary analysis of clinical trial data and primary analysis of medical record data. SETTING: Outpatient clinic. SAMPLE: 149 females and 118 males 12-18 years of age whose cancer had been in remission for at least two years were randomly assigned to intervention (n = 132) and standard care (n = 135) groups. METHODS: Self-report questionnaires, abstracted medical record data, confirmatory factor analysis, and structural equation modeling. MAIN RESEARCH VARIABLES: Smoking, alcohol consumption, knowledge, risk perceptions, motivation, and worry about cancer and treatment effects. FINDINGS: Three factors directly predicted substance use at baseline: being in a higher grade in school (independent of age), feelings of being more susceptible to late effects of cancer therapy, and worrying more about cancer and its treatment. At follow-up a year later, grade in school and worry predicted increased substance use. In addition, a desire to change health behavior, influenced by the intervention and gender, predicted decreased substance use. The mechanism of influence of the intervention was evident: The intervention led to a need to change, which precipitated a desire to change and ultimately resulted in decreased substance use. CONCLUSIONS: Young survivors' worries and concerns about their cancer and treatment-related late effects are a new intervention target. Motivation is sensitive to behavioral change interventions and positively affects risk reduction. IMPLICATIONS FOR NURSING: Two new intervention strategies to address the impact of survivors' concerns about their cancer and its treatment are implied: (a) Replace substance use with new coping methods to reduce fear and anxiety, and (b) tailor motivation-based interventions to age and gender to communicate graphically and realistically to survivors the personal importance of behavioral change in modifying the risks of late effects. PMID- 16470235 TI - Delirium in older adults with cancer: implications for practice and research. AB - PURPOSE/OBJECTIVES: To provide a comprehensive review of the literature and existing evidence-based findings on delirium in older adults with cancer. DATA SOURCES: Published articles, guidelines, and textbooks. DATA SYNTHESIS: Although delirium generally is recognized as a common geriatric syndrome, a paucity of empirical evidence exists to guide early recognition and treatment of this sequelae of cancer and its treatment in older adults. Delirium probably is more prevalent than citations note because the phenomenon is under-recognized in clinical practice across varied settings of cancer care. CONCLUSIONS: Extensive research is needed to formulate clinical guidelines to manage delirium. A focus on delirium in acute care and at the end of life precludes identification of this symptom in ambulatory care, where most cancer therapies are used. Particular emphasis should address the early recognition of prodromal signs of delirium to reduce symptom severity. IMPLICATIONS FOR NURSING: Ongoing assessment opportunities and close proximity to patients' treatment experiences foster oncology nurses' mastery of this common exemplar of altered cognition in older adults with cancer. Increasing awareness of and knowledge delineating characteristics of delirium in older patients with cancer can promote early recognition, optimum treatment, and minimization of untoward consequences associated with the historically ignored example of symptom distress. PMID- 16470236 TI - Expanded treatment options in the adjuvant therapy of colon cancer: implications for oncology nurses. AB - PURPOSE/OBJECTIVES: To review the role of adjuvant therapy in the treatment of patients with colon cancer. DATA SOURCES: Published articles, Internet sources, and books. DATA SYNTHESIS: Colon cancer is a very common cancer in men and women. Chemotherapy, consisting primarily of 5-fluorouracil, has been used to treat colon cancer since the 1950s, but additional effective agents against metastatic disease now are available. The options for adjuvant chemotherapy have increased dramatically. Ongoing studies are evaluating the role of biologics in adjuvant therapy of colon cancer. CONCLUSIONS: Use of oxaliplatin in the adjuvant setting has further defined exciting new therapy options for patients with colon cancer. IMPLICATIONS FOR NURSING: Oncology nurses caring for patients with colon cancer should be aware of new changes in therapy options. Although the addition of new therapies increases the tools in the drug arsenal for the common disease, management of toxicities of therapy is crucial as well. This article reviews changes in therapy options and toxicity management, including discussion of key issues for oncology nurses in the care of patients with colon cancer. PMID- 16470237 TI - Relief of symptoms, side effects, and psychological distress through use of complementary and alternative medicine in women with breast cancer. AB - PURPOSE/OBJECTIVES: To identify use of complementary and alternative medicine (CAM) for relief of symptoms and side effects among women diagnosed with breast cancer and to identify demographic and clinical factors associated with the use of CAM in these patients. DESIGN: A descriptive, cross-sectional survey. SETTING: Clinics and community groups in the Tampa Bay area and community groups in a rural midwestern area. SAMPLE: A convenience sample of 105 predominantly Caucasian women (mean age = 59 years) with a diagnosis of breast cancer was recruited from the Tampa Bay area and a rural midwestern area. METHODS: The instrument used to gather the data was the Use of Complementary Therapies Survey. The reasons for choosing 33 individual CAM treatments were tabulated. The frequency of use was calculated according to four reasons: (a) to reduce physical symptoms or side effects, (b) to reduce psychological distress, (c) to gain a feeling of control over treatment, or (d) because of dissatisfaction with traditional medical care. Least-squares regression models were fit to identify independent demographic and clinical predictors of CAM therapy use. MAIN RESEARCH VARIABLES: Use of CAM for relief of physical and psychological distress. FINDINGS: Patients used all categories of CAM therapies to reduce physical symptoms and side effects. The most frequently cited reason for use of CAM was to reduce the symptom of psychological distress, whereas the lowest frequency of CAM use was because of dissatisfaction with traditional medical care. Traditional and ethnic medicines frequently were used to reduce physical symptoms and side effects, followed by diet and nutritional supplements. The most frequently used CAM therapy category cited for gaining a feeling of control over treatment was use of diet and nutritional supplements. Previous chemotherapy and having more than a high school education were associated with more frequent use of diet and nutritional supplements and stress-reducing techniques. CONCLUSIONS: Frequency of specific use according to type of CAM was higher and more specific than reported in other studies. Patients who had undergone chemotherapy were most likely to use CAM. IMPLICATIONS FOR NURSING: Oncology nurses are in a key position to identify which symptoms or side effects patients are experiencing and which CAM therapies may be helpful to relieve patients' symptoms related to treatment and psychological distress related to their cancer. PMID- 16470238 TI - Sociocultural context of mammography screening use. AB - PURPOSE/OBJECTIVES: To examine variations in cultural and health beliefs about mammography screening among a socioeconomically diverse sample of African American and Caucasian women and to identify which set of beliefs predicted mammography screening adherence. DESIGN: Descriptive, retrospective, cross sectional study. SETTING: Community-based organizations and public housing. SAMPLE: 111 African American women and 64 Caucasian women, aged 40 years or older, with no history of breast cancer. METHODS: Telephone and in-person structured interviews were conducted. Items used previously validated scales based on the Cultural Assessment Model for Health and the Health Belief Model. MAIN RESEARCH VARIABLES: Race or ethnicity, education, income, personal space, health temporal orientation, personal control, fatalism, susceptibility, benefits, barriers, self-efficacy, and mammography screening adherence. FINDINGS: African American women were more fatalistic about breast cancer and perceived fewer benefits to screening. Mammography screening-adherent women were more future oriented, believed that they had less control over finding health problems early, had fewer barriers to screening, and experienced more physical spatial discomfort during the screening procedure than nonadherent women. CONCLUSIONS: Several of the cultural beliefs were not significantly different by race or ethnicity. However, cultural and health beliefs were significant predictors of mammography screening. IMPLICATIONS FOR NURSING: Theoretically based cultural beliefs are important to consider for behavioral interventions to increase mammography screening in African American and Caucasian women. PMID- 16470239 TI - Factors affecting mammography behavior and intention among Korean women. AB - PURPOSE/OBJECTIVES: To understand factors that influence the mammography experience and intention to receive mammography among Korean women using the Health Belief Model and subjective norm of the Theory of Reasoned Action. DESIGN: Cross-sectional. SETTING: Two university hospitals and one general hospital in Korea. SAMPLE: A convenience sample of 310 women aged 30 years and older who visited participating hospitals during the data collection period. METHODS: Self administered questionnaire. MAIN RESEARCH VARIABLES: Perceived susceptibility and severity, perceived benefits and barriers, self-efficacy, normative beliefs, and motivation to comply. FINDINGS: Age, self-efficacy, and perceived susceptibility were significantly associated with participants' mammography experience, whereas knowledge, self-efficacy, perceived susceptibility, perceived barriers, subjective norm, and income were significant in predicting women's intention to receive mammography. CONCLUSIONS: The combined model synthesizing the Health Belief Model and the Theory of Reasoned Action was more effective in predicting mammography intention than in explaining mammography experience. IMPLICATIONS FOR NURSING: Tailored health education and health promotion programs to promote mammography screening among Korean women should be developed based on women's perceptions and norms. PMID- 16470240 TI - Women undergoing breast diagnostics: the lived experience of spirituality. AB - PURPOSE/OBJECTIVES: To explore perceptions of spirituality in women who had undergone a breast diagnostic experience. RESEARCH APPROACH: Qualitative, phenomenologic study using Giorgi's approach. SETTING: An outpatient comprehensive breast assessment center. PARTICIPANTS: 20 Caucasian women, aged 30 89, who had just completed the diagnostic process, including definitive diagnosis, regarding a breast abnormality. METHODOLOGIC APPROACH: In-depth, semistructured, tape-recorded, and transcribed interviews analyzed using the Giorgi method of coding, transforming, and synthesizing data. MAIN RESEARCH VARIABLES: Descriptions of spirituality, spiritual needs, and supporting spirituality. FINDINGS: Two themes emerged: creating a focused isolation and seeking connections. Women created a private mental world in which to concentrate on the stressful diagnostic process. Within the isolation, women explored their personal strength and their connection to God or their spiritual beliefs. When the stress began to overwhelm, they sought out loved ones for support and diversion. Women found the center's staff to be supportive; however, many did not wish to speak to an agency chaplain during the uncertain period. CONCLUSIONS: Women needed to handle the stress alone, with reliance on spirituality and God that was balanced with a need for specific connections to family members or close friends. INTERPRETATION: Nurses can support women's need to focus and can assist family members to understand their role during the diagnostic period. The diagnostic period can be used to talk with women about their spiritual beliefs, their needs, and possible referral to a chaplain. PMID- 16470241 TI - Integrative review of lycopene and breast cancer. AB - PURPOSE/OBJECTIVES: To perform an integrative research analysis of the literature regarding lycopene, its antioxidant capacity, and its nutritive and physiologic effects on breast cancer. DATA SOURCES: Articles published from 1990-2004 using PubMed as the primary retrieval base. DATA SYNTHESIS: Forty articles were retrieved and a dimensional analysis was performed based on Fawcett's integrative review process that focused on the following categories: antioxidant, bioavailability, breast cancer risk, and dietary factors. CONCLUSIONS: Noted gaps in the literature included lack of a theoretical model and multiple nutritional variables studied that affected statistical interpretation and application. Findings suggest that additional research is needed to effectively study the specific phytochemical attributes of lycopene with regard to breast cancer. IMPLICATIONS FOR NURSING: The emerging area of health-derived benefits from food sources such as lycopene requires additional inquiry into the examination of physiological and nutritional parameters. Nurses should include antioxidant therapy in their base of knowledge when caring for patients with breast cancer. PMID- 16470242 TI - Women's perceptions of the effectiveness of telephone support and education on their adjustment to breast cancer. AB - PURPOSE/OBJECTIVES: Social support is believed to be important in helping women adjust to breast cancer. Reports have suggested limited positive effects of social support on well-being, mood disturbances, and relationships with significant others for women who receive telephone support. Women's perceptions of the role of social support in recovery, however, has had limited study. The purpose of this study was to describe women's perceptions of their emotional and interpersonal adaptations to breast cancer after their involvement in a randomized clinical trial in which one group received educational materials and telephone support from oncology nurses and another group received educational materials only. RESEARCH APPROACH: Content analysis was used to discover women's perceptions of their emotional and interpersonal adaptation to breast cancer following their participation in a study in which one group received educational materials and telephone support from oncology nurses and another group received educational materials only. SETTING: All participants were interviewed by telephone in their homes. PARTICIPANTS: 77 of 106 women with breast cancer from a randomized clinical trial were interviewed about their expectations of their adaptations and the effectiveness of the experimental and social support intervention delivered by telephone. METHODOLOGIC APPROACH: Telephone interviews were recorded on audiotape and transcribed for analysis. Structured interviews were completed by a non-nurse interviewer. Frequency counts were obtained from the responses to items and comments were clustered for themes. MAIN RESEARCH VARIABLES: Emotional and interpersonal adaptations to breast cancer, educational materials, and telephone support from oncology nurses. FINDINGS: Fifty-four percent of the women who received the telephone support interventions reported improvement in attitude, whereas 43% of the interviewed women in the control group reported improvement. Only three participants, all in the control group, reported worsened emotional status. The percentage of those reporting improved or unchanged physical status was about equal in each group. The majority of participants in both groups indicated that their levels of involvement in activities remained the same or increased. Forty-six percent of participants in the intervention group reported improved relationships with their spouses compared to 38% in the control group. Women from both groups indicated that the diagnosis of cancer had caused them to review their lives and make changes in their relationships and activities. CONCLUSIONS: Participants who received telephone support for one year, in addition to educational materials, reported improvement in their attitudes toward their breast cancer and better relationships with their significant others. INTERPRETATION: The women's perceptions are consistent with quantitative results from the clinical trial. This article reports additional evidence that telephone support is an effective alternative to support groups and may be appropriate for those with limited access to such groups because of geography, work demands, or family situations. PMID- 16470243 TI - Blocking the alpha 4 integrin-paxillin interaction selectively impairs mononuclear leukocyte recruitment to an inflammatory site. AB - Antagonists to alpha4 integrin show promise for several autoimmune and inflammatory diseases but may exhibit mechanism-based toxicities. We tested the capacity of blockade of alpha4 integrin signaling to perturb functions involved in inflammation, while limiting potential adverse effects. We generated and characterized mice bearing a Y991A mutation in alpha4 integrin [alpha4(Y991A) mice], which blocks paxillin binding and inhibits alpha4 integrin signals that support leukocyte migration. In contrast to the embryonic-lethal phenotype of alpha4 integrin-null mice, mice bearing the alpha4(Y991A) mutation were viable and fertile; however, they exhibited defective recruitment of mononuclear leukocytes into thioglycollate-induced peritonitis. Alpha4 integrins are essential for definitive hematopoiesis; however, the alpha4(Y991A) mice had intact lymphohematopoiesis and, with the exception of reduced Peyer's patches, normal architecture and cellularity of secondary lymphoid tissues. We conclude that interference with alpha4 integrin signaling can selectively impair mononuclear leukocyte recruitment to sites of inflammation while sparing vital functions of alpha4 integrins in development and hematopoiesis. PMID- 16470244 TI - Pericytes limit tumor cell metastasis. AB - Previously we observed that neural cell adhesion molecule (NCAM) deficiency in beta tumor cells facilitates metastasis into distant organs and local lymph nodes. Here, we show that NCAM-deficient beta cell tumors grew leaky blood vessels with perturbed pericyte-endothelial cell-cell interactions and deficient perivascular deposition of ECM components. Conversely, tumor cell expression of NCAM in a fibrosarcoma model (T241) improved pericyte recruitment and increased perivascular deposition of ECM molecules. Together, these findings suggest that NCAM may limit tumor cell metastasis by stabilizing the microvessel wall. To directly address whether pericyte dysfunction increases the metastatic potential of solid tumors, we studied beta cell tumorigenesis in primary pericyte-deficient Pdgfb(ret/ret) mice. This resulted in beta tumor cell metastases in distant organs and local lymph nodes, demonstrating a role for pericytes in limiting tumor cell metastasis. These data support a new model for how tumor cells trigger metastasis by perturbing pericyte-endothelial cell-cell interactions. PMID- 16470245 TI - Elastin fragments drive disease progression in a murine model of emphysema. AB - Mice lacking macrophage elastase (matrix metalloproteinase-12, or MMP-12) were previously shown to be protected from the development of cigarette smoke-induced emphysema and from the accumulation of lung macrophages normally induced by chronic exposure to cigarette smoke. To determine the basis for macrophage accumulation in experimental emphysema, we now show that bronchoalveolar lavage fluid from WT smoke-exposed animals contained chemotactic activity for monocytes in vitro that was absent in lavage fluid from macrophage elastase-deficient mice. Fractionation of the bronchoalveolar lavage fluid demonstrated the presence of elastin fragments only in the fractions containing chemotactic activity. An mAb against elastin fragments eliminated both the in vitro chemotactic activity and cigarette smoke-induced monocyte recruitment to the lung in vivo. Porcine pancreatic elastase was used to recruit monocytes to the lung and to generate emphysema. Elastin fragment antagonism in this model abrogated both macrophage accumulation and airspace enlargement. PMID- 16470246 TI - Loss of IRF-4-binding protein leads to the spontaneous development of systemic autoimmunity. AB - IFN regulatory factor 4-binding (IRF-4-binding) protein (IBP) is a novel type of activator of Rho GTPases that is recruited to the immunological synapse upon TCR stimulation. Here we demonstrate that loss of IBP leads to the spontaneous development of a systemic autoimmune disorder characterized by the accumulation of effector/memory T cells and IgG+ B cells, profound hypergammaglobulinemia, and autoantibody production. Similar to human SLE, this syndrome primarily affects females. T cells from IBP-deficient mice are resistant to death in vitro as well as in vivo and exhibit selective defects in effector function. In the absence of IBP, T cells respond suboptimally to TCR engagement, as demonstrated by diminished ERK1/2 activation, decreased c-Fos induction, impaired immunological synapse formation, and defective actin polymerization. Transduction of IBP deficient T cells with a WT IBP protein, but not with an IBP mutant lacking the Dbl-like domain required for Rho GTPase activation, rescues the cytoskeletal defects exhibited by these cells. Collectively, these findings indicate that IBP, a novel regulator of Rho GTPases, is required for optimal T cell effector function, lymphocyte homeostasis, and the prevention of systemic autoimmunity. PMID- 16470248 TI - Molecular genetics of Alzheimer's disease and aging. AB - Alzheimer's disease is a genetically complex disorder associated with multiple genetic defects, either mutational or of susceptibility. Although potentially associated with an accelerated stochastically driven aging process, Alzheimer's disease is an independent clinical entity in which the aging process exerts a deleterious effect on brain activity in conjunction with polymodal genetic factors and other pathological conditions (i.e., age-related cerebrovascular deterioration) and environmental factors (i.e., nutrition). Alzheimer's disease genetics does not explain in full the etiopathogenesis of this disease. Therefore, it is likely that environmental factors and/or epigenetic phenomena also contribute to Alzheimer's disease pathology and phenotypic expression of dementia. The genomics of Alzheimer's disease is still in its infancy, but this field is aiding the understanding of novel aspects of this disease, including genetic epidemiology, multifactorial risk factors, pathogenic mechanisms associated with genetic networks and genetically regulated metabolic cascades. Alzheimer's disease genomics is also helping to develop new strategies in pharmacogenomic research and prevention. Functional genomics, proteomics, pharmacogenomics, high-throughput methods, combinatorial chemistry and modern bioinformatics will greatly contribute to accelerate drug development for Alzheimer's disease and other complex disorders. The multifactorial genetic dysfunction in dementia includes mutational loci (APP, PS1, PS2, TAU) and diverse susceptibility loci (APOE, alpha2M, alphaACT, LRP1, IL1 alpha, TNF, ACE, BACE, BCHE, CST3, MTHFR, GSK3 beta, NOS3 and many other genes) distributed across the human genome, probably converging in a common pathogenic mechanism that leads to premature neuronal death, in which mitochondrial DNA mutations may contribute to increased genetic variability and heterogeneity. In Alzheimer's disease, multiple pathogenic events, including genetic factors, accumulation of aberrant or misfolded proteins, protofibril formation, ubiquitin-proteasome system dysfunction, excitotoxic reactions, oxidative and nitrosative stress, mitochondrial injury, synaptic failure, altered metal homeostasis, dysfunction of axonal and dendritic transport, and chaperone misoperation may converge in pathogenic pathways leading to premature death and neurodegeneration. Some of these mechanisms are common to several neurodegenerative disorders, which differ depending upon the gene(s) affected and the involvement of specific genetic networks, together with epigenetic factors and environmental events. Many genes potentially associated with Alzheimer's disease in some studies cannot be confirmed as candidate genes in replication studies, indicating that methodological problems and genomic complexity are leading to erroneous conclusions. A different approach to Alzheimer's disease functional genomics is to integrate individual genetic information in polygenic genotypes (haplotype like model) and to investigate genotype-phenotype correlations and genotype related pharmacogenomic behaviors. The application of functional genomics to Alzheimer's disease can be a suitable strategy for molecular diagnosis and for understanding pathophysiological mechanisms associated with Alzheimer's disease related neurodegeneration. Furthermore, the pharmacogenomics of Alzheimer's disease may contribute in the future to optimize drug development and therapeutics, increasing efficacy and safety, and reducing side-effects and unnecessary costs. PMID- 16470247 TI - CFTR is required for PKA-regulated ATP sensitivity of Kir1.1 potassium channels in mouse kidney. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel plays vital roles in fluid transport in many epithelia. While CFTR is expressed along the entire nephron, its function in renal tubule epithelial cells remains unclear, as no specific renal phenotype has been identified in cystic fibrosis. CFTR has been proposed as a regulator of the 30 pS, ATP-sensitive renal K channel (Kir1.1, also known as renal outer medullar K [ROMK]) that is critical for K secretion by cells of the thick ascending limb (TAL) and distal nephron segments responsive to aldosterone. We report here that both ATP and glibenclamide sensitivities of the 30 pS K channel in TAL cells were absent in mice lacking CFTR and in mice homozygous for the deltaF508 mutation. Curcumin treatment in deltaF508-CFTR mice partially reversed the defect in ATP sensitivity. We demonstrate that the effect of CFTR on ATP sensitivity was abrogated by increasing PKA activity. We propose that CFTR regulates the renal K secretory channel by providing a PKA-regulated functional switch that determines the distribution of open and ATP-inhibited K channels in apical membranes. We discuss the potential physiological role of this functional switch in renal K handling during water diuresis and the relevance to renal K homeostasis in cystic fibrosis. PMID- 16470251 TI - Internet-based asthma education - a novel approach to compliance: a case report. AB - Asthma costs Canadians over 1.2 billion dollars per annum and, despite advances, many asthmatic patients still have poor control. An action plan, symptom diary and measurement of peak expiratory flow have been shown to improve clinical outcomes. Effective educational interventions are an important component of good care. However, many rural sites lack not only access to education but physician care as well. It is reasonable, therefore, that an Internet-based asthma management program may be used as an approach. In the present case report, a novel approach that may increase access in these poorly serviced areas is presented. In an Internet-based asthma management program, patients are reviewed by a physician, receive education and are given a unique password that provides program access. Patients record symptoms and peak expiratory flow rates. The present case report shows that a patient can be assisted through an exacerbation, thus averting emergency intervention and stabilizing control, even when travelling on another continent. PMID- 16470249 TI - Risk of ruling out severe acute respiratory syndrome by ruling in another diagnosis: variable incidence of atypical bacteria coinfection based on diagnostic assays. AB - BACKGROUND: Severe acute respiratory syndrome (SARS) caused the first epidemic of the 21st century and continues to threaten the global community. OBJECTIVE: To assess the incidence of coinfection in patients confirmed to have SARS-associated coronavirus (SARS-CoV) infection, and thus, to determine the risk of ruling out SARS by ruling in another diagnosis. METHODS: The present report is a retrospective study evaluating the incidence and impact of laboratory-confirmed SARS-CoV and other pulmonary pathogens in 117 patients. These patients were evaluated in a Toronto, Ontario, community hospital identified as the epicentre for the second SARS outbreak. RESULTS: Coinfection with other pulmonary pathogens occurred in patients with SARS. Seventy-three per cent of the patient population evaluated had laboratory-confirmed SARS-CoV infection. Serology showing acute or recent Chlamydophila pneumoniae or Mycoplasma pneumoniae infection revealed an incidence of 30% and 9%, respectively, in those with SARS. These rates are similar to previously published studies on coinfection in pneumonia. All nucleic acid diagnostic assays were negative for C pneumoniae and M pneumoniae in respiratory samples from patients with SARS having serological evidence for these atypical pathogens. CONCLUSIONS: Diagnostic assays for well-recognized pulmonary pathogens have limitations, and ruling out SARS-CoV by ruling in another pulmonary pathogen carries significant risk. Despite positive serology for atypical pathogens, in a setting where clinical suspicion for SARS is high, specific tests for SARS should be performed to confirm or exclude a diagnosis. PMID- 16470252 TI - Small wonders, big future: the development of environmental nanotechnology. AB - Nanotechnology--the engineering of atomic-scale devices and systems--is set to become one of the major industries of the 21st century. Environmental applications are emerging as a key area of interest. But alongside the benefits, there are concerns that nanotechnologies could bring new risks to the environment and human health. Are nanotechnologies friend or foe? And how can analysts contribute to the debate? PMID- 16470256 TI - Simultaneous measurement of ventilation using tracer gas techniques and VOC concentrations in homes, garages and vehicles. AB - Air exchange rates and interzonal flows are critical ventilation parameters that affect thermal comfort, air migration, and contaminant exposure in buildings and other environments. This paper presents the development of an updated approach to measure these parameters using perfluorocarbon tracer (PFT) gases, the constant injection rate method, and adsorbent-based sampling of PFT concentrations. The design of miniature PFT sources using hexafluorotoluene and octafluorobenzene tracers, and the development and validation of an analytical GC/MS method for these tracers are described. We show that simultaneous deployment of sources and passive samplers, which is logistically advantageous, will not cause significant errors over multiday measurement periods in building, or over shorter periods in rapidly ventilated spaces like vehicle cabins. Measurement of the tracers over periods of hours to a week may be accomplished using active or passive samplers, and low method detection limits (<0.025 microg m(-3)) and high precisions (<10%) are easily achieved. The method obtains the effective air exchange rate (AER), which is relevant to characterizing long-term exposures, especially when ventilation rates are time-varying. In addition to measuring the PFT tracers, concentrations of other volatile organic compounds (VOCs) are simultaneously determined. Pilot tests in three environments (residence, garage, and vehicle cabin) demonstrate the utility of the method. The 4 day effective AER in the house was 0.20 h(-1), the 4 day AER in the attached garage was 0.80 h(-1), and 16% of the ventilation in the house migrated from the garage. The 5 h AER in a vehicle traveling at 100 km h(-1) under a low-to-medium vent condition was 92 h( 1), and this represents the highest speed test found in the literature. The method is attractive in that it simultaneously determines AERs, interzonal flows, and VOC concentrations over long and representative test periods. These measurements are practical, cost-effective, and helpful in indoor air quality and other investigations. PMID- 16470257 TI - The use of semipermeable membrane devices as passive samplers to determine persistent organic compounds in indoor air. AB - In the study reported here semipermeable membrane devices (SPMDs) were used to sample 28 PAHs and 19 PCBs in the gas phase in 15 single-family houses located in an area where domestic wood burning is widespread. Eight of the households used wood burning appliances whereas the others used other systems for residential heating. Most of the studied compounds were found in the houses: the PAHs at levels that were similar to or slightly higher than published SPMD-sampled levels for background or urban sites in Sweden, and the PCBs at levels that were somewhat lower than those recently found in both indoor and outdoor urban locations. A principal component analysis revealed that wood-burning heating systems may contribute to PAHs in indoor air. The sources may be emissions indoors or penetration from outdoors. The convenience of SPMD technology facilitates its use for semi-quantitative screening and monitoring of various persistent organic compounds indoors in dwellings and working environments. PMID- 16470258 TI - Comparison of 24 h averaged VOC monitoring results for residential indoor and outdoor air using Carbopack X-filled diffusive samplers and active sampling--a pilot study. AB - Analytical results obtained by thermal desorption GC/MS for 24 h diffusive sampling of 11 volatile organic compounds (VOCs) are compared with results of time-averaged active sampling at a known constant flow rate. Air samples were collected with co-located duplicate diffusive sampling tubes and one passivated canister. A total of eight multiple-component sampling events took place at fixed positions inside and outside three private homes. Subsequently, a known amount of sample air was transferred from the canister to an adsorbent tube for analysis by thermal desorption GC/MS. Results for the 11 most prevalent compounds--Freon 11, 1,3-butadiene, benzene, toluene, tetrachloroethene, ethylbenzene, m,p-xylene, o xylene, 4-ethyltoluene, 1,3,5-trimethylbenzene, and p-dichlorobenzene--show that the ratio of average study values (diffusive sampling to active sampling) is 0.92 with 0.70 and 1.14 extreme ratios. Absolute percent difference for duplicate samples using diffusive sampling was <10% for the four most prevalent compounds. Agreement between the two sampling approaches indicates that the prediction of approximately constant diffusive sampling rates based on previous laboratory studies is valid under the field conditions. PMID- 16470250 TI - Factors associated with poor asthma control in children aged five to 13 years. AB - BACKGROUND: Most children with asthma should be able to achieve acceptable control. However, are there differences between those with acceptable and poor control, and if so, how can health care approaches be modified accordingly? OBJECTIVE: To examine the characteristics of elementary school children aged five to 13 years with acceptable and poor levels of asthma control. METHODS: The present cross-sectional study of children with asthma used five indicators of control, as outlined by the Canadian Asthma Consensus Report, to categorize acceptable and poor asthma control. RESULTS: Of 153 children, 115 (75%) were rated as having poorly controlled asthma. Of those with poor control, 65 (64%) children were currently using inhaled corticosteroids, and 65% of those reported using inhaled corticosteroids daily versus as needed. Fifty-one per cent of the children with poorly controlled asthma had exposure to tobacco smoke, whereas 79% of the children with asthma under acceptable control were from households with no smokers (P=0.002). The poor control group also had significantly worse parental perceptions of the psychosocial impact of asthma on their child. No significant difference was found in the percentage of those who had written action plans in the poor control group (28%) compared with the acceptable control group (26%), and similar percentages in each group stated that they used the plans. CONCLUSIONS: Despite the high use of inhaled corticosteroids, the majority of children had poorly controlled asthma. The poor control group had more exposure to tobacco smoke and a worse psychosocial impact due to asthma. Few children had past asthma education and action plans, suggesting that there is a need to improve access to and tools for education. PMID- 16470259 TI - Chamber evaluation of a portable GC with tunable retention and microsensor-array detection for indoor air quality monitoring. AB - The evaluation of a novel prototype instrument designed for on-site determinations of VOC mixtures found in indoor working environments is described. The instrument contains a miniature multi-stage preconcentrator, a dual-column separation module with pressure-tunable retention capabilities, and an integrated array of three polymer-coated surface acoustic wave sensors. It was challenged with dynamic test-atmospheres of a set of 15 common indoor air contaminants at parts-per-billion concentrations within a stainless-steel chamber under a range of conditions. Vapours were reliably identified at a known level of confidence by combining column retention times with sensor-array response patterns and applying a multivariate statistical test of pattern fidelity for the chromatographically resolved vapours. Estimates of vapour concentrations fell within 7% on average of those determined by EPA Method TO-17, and limits of detection ranged from 0.2 to 28 ppb at 25 degrees C for 1 L samples collected and analyzed in <12 min. No significant humidity effects were observed (0-90% RH). Increasing the chamber temperature from 25 to 30 degrees C reduced the retention times of the more volatile analytes which, in turn, demanded alterations in the scheduling of column-junction-point pressure (flow) modulations performed during the analysis. Reductions in sensor sensitivities with increasing temperature were predictable and similar among the sensors in the array such that most response patterns were not altered significantly. Short-term fluctuations in concentration were accurately tracked by the instrument. Results indicate that this type of instrument could provide routine, semi-autonomous, near-real-time, multi-vapour monitoring in support of efforts to assess air quality in office environments. PMID- 16470260 TI - An indoor test campaign of the tomography long path differential optical absorption spectroscopy technique. AB - In this study we validate the two-dimensional long path DOAS tomography measurement technique by means of an indoor experiment with well-known concentration distributions. The experiment was conducted over an area of 10 m x 15 m using one and two cylindrical polycarbonate containers of diameter 2 m, respectively, filled with NO2. The setup was realized with three of the multibeam instruments recently developed by Pundt and Mettendorf (Appl. Opt., 2005, in press), which allow the simultaneous measurement along at least four light paths each. The configuration consisted of twelve simultaneous light beams, 39 horizontal light paths in total, and 18 different cylinder positions inside the field. It was found that for the discretization and inversion technique shown here reconstructions of the concentration distributions from experimental data agree well with simulated reconstructions. In order to draw conclusions for atmospheric applications, numerical studies including instrumental errors were carried out. It was found that with the presented measurement setup it is possible to measure and reconstruct one or two NO2 plumes of 600 m diameter and average concentrations above 4.2 ppbv each, on a scale of 13.5 km2. Theoretical investigations show that it should be possible to localize and quantify 600 m diameter plumes of SO2 > 1.5 ppbv, H2CO > 6.3 ppbv, HONO > 3.2 ppbv, and ozone > 46.2 ppbv. Larger plumes can be measured with higher precision. PMID- 16470261 TI - Contamination of Canadian and European bottled waters with antimony from PET containers. AB - Using clean lab methods and protocols developed for measuring Sb in polar snow and ice, we report the abundance of Sb in fifteen brands of bottled water from Canada and forty-eight from Europe. Comparison with the natural abundance of Sb in pristine groundwaters, water bottled commercially in polypropylene, analyses of source waters prior to bottling, and addition of uncontaminated groundwater to PET bottles, provides unambiguous evidence of Sb leaching from the containers. In contrast to the pristine groundwater in Ontario, Canada containing 2.2 +/- 1.2 ng l(-1) Sb, 12 brands of bottled natural waters from Canada contained 156 +/- 86 ng l(-1) and 3 brands of deionized water contained 162 +/- 30 ng l(-1); all of these were bottled in PET containers. Natural water from Ontario bottled in polypropylene contained only 8.2 +/- 0.9 ng l(-1). Comparison of three German brands of water available in both glass bottles and PET containers showed that waters bottled in PET contained up to 30 times more Sb. To confirm that the elevated Sb concentrations are due to leaching from the PET containers, water was collected in acid-cleaned LDPE bottles from a commercial source in Germany, prior to bottling; this water was found to contain 3.8 +/- 0.9 ng l(-1) Sb (n = 5), compared with the same brand of water purchased locally in PET bottles containing 359 +/- 54 ng l(-1) (n = 6). This same brand of water in PET bottles, after an additional three months of storage at room temperature, yielded 626 +/- 15 ng l( 1) Sb (n = 3). Other German brands of water in PET bottles contained 253-546 ng l(-1) Sb (n = 5). The median concentration of Sb in thirty-five brands of water bottled in PET from eleven other European countries was 343 ng l(-1) (n = 35). As an independent check of the hypothesis that Sb is leaching from PET, the pristine groundwater from Canada (containing 2.2 +/- 1.2 ng l(-1) Sb) was collected from the source using PET bottles from Germany: this water contained 50 +/- 17 ng l( 1) Sb (n = 2) after only 37 days, even though it was stored in the refrigerator, and 566 ng l(-1) after six months storage at room temperature. PMID- 16470262 TI - Urinary 8-hydroxy-2'-deoxyguanosine in inhabitants chronically exposed to arsenic in groundwater in Cambodia. AB - Arsenic concentrations in hair and urine, and urinary levels of 8-hydroxy-2' deoxyguanosine (8-OHdG), a marker of oxidative DNA damage, were examined for inhabitants of the Mekong Basin in Kratie Province, Cambodia. Also, the arsenic levels of tube-well water were determined. Total arsenic concentrations in tube well water ranged from <1 to 886 microg L(-1), and 44.8% of these exceeded the WHO drinking water guideline of 10 microg L(-1). Elevated levels of arsenic were observed in the human hair and urine, and also a significant positive correlation was observed between the concentrations in hair and urine. These results suggest that the inhabitants are chronically exposed to arsenic through drinking the tube well water. Levels of urinary 8-OHdG were higher for the subjects with higher arsenic levels in hair and urine, suggesting that induction of oxidative DNA damage was caused by chronic exposure to arsenic in tube-well water for the inhabitants in Kratie Province. To our knowledge, this is the first report on the oxidative DNA damage caused by chronic exposure to arsenic in groundwater for the inhabitants in Cambodia. PMID- 16470263 TI - Characterization and diurnal variation of size-resolved inorganic water-soluble ions at a rural background site. AB - Water-soluble inorganic ions in aerosol samples have been studied. The sample collection took place during summer in 2003 at a European background site which is operating within the framework of the European Monitoring and Evaluation Program. Gent type PM10 stacked filter unit (SFU) samplers were operated in parallel on a day and night basis to collect particles in separate coarse (2.0-10 microm) and fine (<2.0 microm) size fractions. Particulate masses were measured gravimetrically; the filters from one of the SFU samplers were analyzed by particle-induced X-ray emission spectrometry (PIXE) and instrumental neutron activation analysis (INAA). Filters from the other SFU sampler were analyzed by ion chromatography (IC) for major inorganic anions (MSA-, NO2(-), NO3(-), Cl-, Br , SO4(2-), oxalate) and cations (Na+, K+, NH4(+), Mg2+, Ca2+). The water-soluble inorganic ions measured were responsible for 44% and 16% of the total fine and coarse particulate mass, respectively. In the fine size fraction, the main ionic components were SO4(2-) and NH4(+) accounting for about 90% of fine ionic mass. In the coarse fraction the main ionic components were Ca2+ and NO3(-), followed by SO4(2-). Significant day and night difference in the mass concentrations was observed only for fine NO3(-). The molar ratios of fine NH4(+) to SO4(2-) indicated their complete neutralization to (NH4)2SO4. According to the cation-to anion ratios the coarse particles were alkaline, while the fine particles were slightly acidic or neutral. By comparing the corresponding concentrations obtained from PIXE/INAA and IC, we determined the water-extractable part of the individual species. We also investigated the effect of long-range transported air masses on the local air concentrations, and we found that the air quality of this background monitoring station was affected by regional pollution sources. PMID- 16470264 TI - The distribution and composition of hydrocarbons in sediments from the Fladen Ground, North Sea, an area of oil production. AB - The distribution and composition of hydrocarbons in sediment from the Fladen Ground oilfield in the northern North Sea have been investigated. The total PAH concentrations (2- to 6-ring parent and alkylated PAHs, including the 16 US EPA PAHs) in sediments were relatively low (<100 microg kg(-1) dry weight). The PAH, the Forties crude and diesel oil equivalent concentrations were generally higher in sediment of fine grain size and higher organic carbon concentration. PAH distributions and concentration ratios indicated a predominantly pyrolytic input, being dominated by the heavier, more persistent, 5- and 6-ring compounds, and with a high proportion of parent PAHs. The n-alkane profiles of a number of the sediments contained small, high boiling point, UCMs, indicative of weathered oil arising from a limited petrogenic input. The geochemical biomarker profiles of the sediments that contained UCMs showed a small bisnorhopane peak and a high proportion of norhopane relative to hopane, indicating that there was contamination from both Middle Eastern and North Sea oils. Therefore contamination was not directly as a result of oil exploration activity in the area. The most likely source of petrogenic contamination was from general shipping activity. PMID- 16470265 TI - Towards 2020 vision in Singapore. PMID- 16470266 TI - Therapeutic hypothermia for neonatal hypoxic-ischaemic encephalopathy. PMID- 16470267 TI - The clinical predictors of hypertension and sleepiness in an Asian population with sleep-disordered breathing. AB - INTRODUCTION: The objective of this study was to identify the clinical predictors for hypertension and sleepiness in an Asian population with sleep-disordered breathing (SDB). MATERIALS AND METHODS: This is a retrospective, consecutive case series of 228 patients with symptoms of SDB referred to a tertiary level sleep disorders unit. A full sleep history, body mass index (BMI), Epworth Sleepiness Score (ESS), apnoea-hypopnoea index (AHI) and lowest oxygen saturation were recorded. All patients had an in-hospital polysomnogram. AHI > or =5 defined SDB. ESS >8 defined sleepiness. RESULTS: A BMI of 25 had a positive and negative predictive value of 75% and 56%, respectively, for predicting AHI > or =5, area ROC (receiver operating curve) = 0.668. Patients with AHI >5 had 3 times the risk of developing hypertension compared to the population with AHI <5. Using stepwise multivariate analyses with constant, age, BMI and lowest oxygen desaturation were predictors for hypertension. Patients with AHI > or =5 were 1.88 times more likely to have ESS >8. After stepwise multivariate analyses, the subjective complaint of sleepiness and lowest oxygen saturation during sleep were predictors of ESS >8. CONCLUSION: The lowest oxygen saturation attained during sleep was the common factor for hypertension and sleepiness in our Asian SDB population. Mild obesity predisposes to AHI > or =5 in our population, the threshold at which the risk of hypertension is 3-fold compared with AHI <5. The subjective complaint of sleepiness predicted an ESS of >8. PMID- 16470268 TI - Clinical experience with three combination regimens for the treatment of high risk febrile neutropenia. AB - INTRODUCTION: The objective of this study was to compare the safety and efficacy of ceftazidime (2 g every 8 h), piperacillin/tazobactam (4 g/500 mg every 6 h), and meropenem (1 g every 8 h), when combined with amikacin (15 mg/kg once daily), in the empirical treatment of high-risk febrile neutropenic episodes in patients with haematological malignancy. MATERIALS AND METHODS: A prospective, comparative study designed in the haematology unit of a university hospital in Turkey. RESULTS: A total of 89 febrile episodes in 60 neutropenic patients were treated; 29 febrile episodes in 23 patients with ceftazidime plus amikacin (group 1), 30 episodes in 25 patients with piperacillin/tazobactam plus amikacin (group 2), and 30 episodes in 25 patients with meropenem plus amikacin (group 3). The 3 groups were comparable in terms of age, sex, underlying malignancy, pretherapy neutrophil counts, duration of neutropenia and types of infections. Neutropenia, since the start of fever, persisted for > or =10 days in all of the episodes in the 3 study groups. Nearly all of the episodes were seen in patients with acute leukaemia. In 25.8% (23/89) of the febrile neutropenia episodes, an aetiologic organism was isolated, with gram-negative bacteria being the most commonly isolated. The success without modification rates were 34.5%, 30% and 36.7% for groups 1, 2 and 3, respectively (P >0.05). After modification with a different class of antimicrobial therapy, the response rates increased to 65.5%, 63.3% and 70% for groups 1, 2 and 3, respectively (P >0.05). The mean duration of treatment and the time to defervescence were also comparable in all groups. In all arms, side effects were minimal. CONCLUSIONS: It is concluded that the 3 regimens were equally effective and safe in the empirical treatment of high-risk febrile neutropenic episodes. PMID- 16470269 TI - Factors associated with functional decline of hospitalised older persons following discharge from an acute geriatric unit. AB - INTRODUCTION: Older persons are likely to develop functional impairment following hospitalisation. Several studies in the West have examined the factors associated with functional decline following the older person's discharge from hospital but there are little data on Asian populations. This study aims to look at the associated risk factors in our local population, following admission to an acute geriatric unit. MATERIALS AND METHODS: This is a retrospective, cohort study. Patients who were discharged from an inpatient geriatric unit over a 3-month period were recruited. Data including their demographic information, functional status prior to admission and at the time of discharge, and medical conditions were obtained from the inpatient medical notes. A follow-up telephone interview was conducted at 3 months to determine the functional status of these patients at that point in time. RESULTS: Following hospitalisation, 40.4% of patients developed functional decline. Of those discharged, 29.6% showed functional decline at 3 months. The principal diagnosis, hypoalbuminaemia, tendency to fall, premorbid functional independence and the length of hospitalisation were associated with functional decline during hospitalisation, while hypoalbuminaemia, the presence of bedsores, institutionalisation, the length of hospitalisation and premorbid functional dependence were important factors associated with functional decline between the time of discharge and 3 months after. In the multivariable predictive model, independent predictors of functional decline at the time of discharge included patient's tendency to fall, premorbid functional independence and the length of hospitalisation, while the presence of bedsores was the only significant predictor of functional decline 3 months post-discharge. CONCLUSIONS: Many elderly patients developed new functional impairment following hospitalisation. Several factors were found to be associated with this functional decline, though no single predictive model similar to the other published studies was identified. PMID- 16470270 TI - Clinical and social correlates of duration of untreated psychosis in first episode psychosis patients. AB - INTRODUCTION: This study aims to examine the socio-demographic and clinical correlates of the duration of untreated psychosis (DUP) in first-episode psychosis patients in an Asian country. MATERIAL AND METHODS: Three hundred thirty-four patients from the Early Psychosis Intervention Programme (EPIP) of Singapore were recruited for the study. Socio-demographic data were collected and patients were diagnosed using SCID I (Structural Clinical Interview for DSM-IV Axis I Disorders). Other assessment scales were used to assess the level of psychopathology, overall functioning and the awareness of mental illness. RESULTS: Mean (SD) DUP was 16.3 (31.5) months. Patients who were single, unemployed, or brought by the police had a significantly longer DUP. Patients with a diagnosis of schizophrenia had a longer DUP as compared to patients with other forms of psychosis. The better functioning patients as indicated by a higher Global Assessment of Functioning score and those who were more insightful had a shorter DUP. CONCLUSIONS: This study suggests that certain socio demographic features and clinical diagnosis may determine DUP. PMID- 16470271 TI - Factors causing delay in discharge of elderly patients in an acute care hospital. AB - INTRODUCTION: Prolonged hospitalisation not only increases cost, it is also associated with other complications. Length of stay (LOS) is one of the indicators that reflect total cost of care during hospitalisation. So, it is of paramount importance to find out why elderly patients overstay in acute care hospitals and address these issues proactively. Since no local data is available, a study was planned to ascertain reasons why the discharging of elderly patients from hospital is delayed and whether these reasons are avoidable. MATERIALS AND METHODS: Long-stay patients were defined as those whose LOS had notably exceeded the average LOS for the Diagnosis Related Group (DRG), based on principal admitting diagnosis. A separate analysis showed that the specialty-specific long stay marker for geriatric medicine was 28 days, so casenote review was done for all patients with LOS of more than 28 days who were discharged from geriatric medicine service of an acute care hospital during a 1-year study period. Information was collected on demographic profile, functional and cognitive status, past medical and social history, admitting medical diagnoses, discharge limiting and delaying factors. RESULTS: During the study period, 150 patients stayed over 28 days and 137 casenotes were available for review. The mean age of the patients was 84 years, 55.5% were female, 77.4% were Chinese, mean abbreviated mental test (AMT) score was 3 and mean modified Barthel's score was 11. The commonest primary diagnosis was sepsis followed by neurological problems, falls-related complication and cardiovascular diseases. The 2 most common discharge limiting factors (the final event which resulted in delay in discharge) were social issues (54, 39.4%) and sepsis (47, 34.3%). Of 47 patients with sepsis, 37 (78.7%) were nosocomial infection. Urinary tract infection and pneumonia were the 2 most common nosocomial infections. The 4 most common factors contributing to delayed discharge (various problems that surfaced throughout the hospital stay) were sepsis (94, 68.6%), deconditioning (65, 47.4%), social issues (52, 38.0%) and cardiovascular disorders (37, 27.0%). CONCLUSION: Elderly patients are more prone to hospitalisation-related complications like nosocomial infection and deconditioning leading to prolonged hospital stay. Early interventions can reduce these complications. Early identification of social issues and prompt discharge planning should be done to avoid delay in discharge. PMID- 16470273 TI - A rotavirus vaccine for infants: the Asian experience. AB - INTRODUCTION: Severe rotavirus gastroenteritis in children causes significant morbidity worldwide and substantial deaths in developing countries. Hence, a live attenuated vaccine Rotarix was developed with human strain RIX4414 of G1P1A P[8] specificity. RIX4414 trials in infants have begun in developed and developing countries worldwide. An overview of RIX4414 in developed and developing countries and prospects with this vaccine in Asia are presented. METHODS: Completed RIX4414 trials have been reviewed. RESULTS: Two oral doses of RIX4414 were well tolerated with a reactogenicity profile similar to placebo. RIX4414 was also highly immunogenic, e.g., in a dose-ranging study conducted in Singapore, 98.8% to 100% of infants had a vaccine take after 2 doses. RIX4414 did not affect the immune response of simultaneously administered routine infant vaccines. RIX4414 significantly reduced severe rotavirus gastroenteritis in settings where multiple serotypes including the emerging G9 type co-circulated. CONCLUSION: These encouraging results warrant further evaluation of the vaccine worldwide and especially in developing countries with the highest need. Therefore, evaluation of the Rotarix vaccine is continuing in large phase III trials in Asia and worldwide. PMID- 16470272 TI - Phase II trial of gemcitabine and cisplatin sequentially administered in Asian patients with unresectable or metastatic non-small cell lung cancer. AB - INTRODUCTION: The aim of this study was to assess toxicity and response in the sequential administration of gemcitabine followed by cisplatin in unresectable or metastatic non-small cell lung cancer. MATERIALS AND METHODS: Twenty-three patients were enrolled in this study. Gemcitabine was given at 1,250 mg/m2 on days 1 and 8, for four 21-day cycles. RESULTS: There were 4 patients with partial responses. 5 patients with stable disease and 10 patients with progressive disease, giving a response rate of 21%. The median time to disease progression was 3.3 months. The median overall survival was 14.6 months. Toxicities graded 3 or 4 included anaemia (13.0%), neutropaenia (13.0%), supraventricular tachycardia (4.3%), and nausea and vomiting (4.3%). CONCLUSION: Although these results show similar efficacy to single-agent treatment regimens, the low toxicity profile and promising survival outcome with this regimen are important points for consideration. PMID- 16470274 TI - A case of cystic schwannoma of the lesser sac. AB - INTRODUCTION: Benign cystic schwannoma of the lesser sac is an extremely rare condition and only 3 reports were found in our review of the English literature. CLINICAL PICTURE: We describe a 58-year-old man with a large (5.2 x 6.7 x 7.6 cm) epigastric mass with solid and cystic components detected on sonography and computed tomography. Close association with the pancreas, stomach and liver led to a mistaken diagnosis of pancreatic cystadenoma/cystadenocarcinoma. TREATMENT AND OUTCOME: The mass was surgically resected and our patient has been well since. CONCLUSION: This case draws the reader's attention to a rare condition that may mimic other sinister lesions, and highlights the pitfalls of diagnosis. Depending on the size and extent of the lesion, imaging may assist in characterisation of a schwannoma of the lesser sac. Surgical resection would still be the end point of management, especially when mass effect causes debilitating symptoms. Surgery is usually associated with good outcome, and tumour recurrence is unusual, provided adequate margins are resected. PMID- 16470275 TI - Swallowed foreign bodies in children: report of four unusual cases. AB - INTRODUCTION: Although a majority of ingested foreign bodies (FBs) pass down the gastrointestinal tract spontaneously, those that are sharp, pointed or large in size need removal to avert serious complications. We highlight the urgent need and utility of endoscopic accessories and technical artistry in safe retrieval of FBs in children. CLINICAL PICTURE: Four children had accidentally swallowed a nail, metallic dumbbell, open safety pin and a cushion pin respectively. They were symptom-free and the abdominal plain radiographs revealed foreign body in the stomach in all the cases. TREATMENT: Oesophago-gastro-duodenoscopy (OGD) was done in all the patients and could retrieve the nail, metallic dumbbell and open safety pin successfully using a Dormia basket, a polypectomy snare and a pair of rat-tooth forceps respectively. The cushion pin had migrated to the duodeno jejunal junction within 4 hours of ingestion and necessitated open duodenotomy and retrieval. OUTCOME: All patients did well after the procedure with no complications. CONCLUSIONS: Swallowed FBs with pointed or sharp ends or large enough to cross the pylorus and duodenal sweep need removal and in the majority of the cases they can be retrieved by OGD. Sharp or pointed FBs that have crossed the second part of the duodenum necessitate urgent laparotomy for retrieval to prevent complications. PMID- 16470276 TI - Clinical cases seen in tsunami hit Banda Aceh: from a primary health care perspective. AB - The 2004 Indian Ocean earthquake and tsunami caused catastrophic damage to many cities on the rim of the Indian Ocean. Banda Aceh in Sumatra, Indonesia was particularly badly hit due to its close proximity to the epicenter. The Singapore Armed Forces Medical Team was one of the earliest medical teams to arrive in Banda Aceh, providing primary health care to the survivors. In the first 17 days of its operation, more than 2000 injured and sick were seen by the Medical Team at 2 locations within Banda Aceh. Approximately one-third of the patients suffered from infected superficial wounds on their limbs and faces. Many developed deep-seated necrosis of the skin tissue, requiring repeated wound dressing and debridement. Another one-third suffered from respiratory tract infection, some due to aspiration of sea water. There were relatively few major trauma and fracture cases seen, and there were a few cases of compression barotrauma of the tympanic membranes resulting from underwater immersion in the sea water. The casualty patterns seem consistent with those reported in other tsunami disasters, which differ distinctly from those seen in a pure earthquake disaster. PMID- 16470278 TI - [Nutritional intervention and primary prevention of type 2 diabetes mellitus: a systematic review]. AB - Type 2 diabetes mellitus is one of the most important public health problems in the world. Its increasing prevalence in several countries and the difficult metabolic control of individuals with the disease justify studying strategies for primary prevention. Recent randomized clinical trials suggest that nutritional intervention may have an important impact on risk reduction for type 2 diabetes mellitus among individuals at high risk. The present systematic review describes the effect of lifestyle intervention trials on primary prevention of type 2 diabetes mellitus. PMID- 16470279 TI - [Hospital Information Systems and their application in public health]. AB - The objective of this study was to survey the scientific production of applications for data from the Hospital Information System of Brazil's Unified National System (SUS) in analyses of relevant Public Health issues. To find articles published in scientific journals from 1984 to 2003, the authors consulted the SciELO, MEDLINE, and Virtual Public Health Library databases. To locate monographs, theses, and dissertations, the authors consulted the websites of institutions providing Master's and doctoral courses in Public Health. A total of 76 articles were identified for the reference period, and they were classified into five categories with different analytical approaches. Although the Hospital Information System in the SUS has incomplete coverage and there are uncertainties about the reliability of its data, the range of studies showed internal consistency with current knowledge, reinforcing the system's importance and the need to understand its strengths and weaknesses. PMID- 16470280 TI - [Social representations by health professionals of sexual violence against women: a study in three municipal public maternity hospitals in Rio de Janeiro, Brazil]. AB - The purpose of this study was to analyze social representations of sexual violence against women, as constructed and reproduced in prenatal care settings in three municipal maternity hospitals in Rio de Janeiro, Brazil. This qualitative research explored two themes: ideas about and explanations of sexual violence committed against woman. The forty-five interviews conducted with health professionals were examined using thematic content analysis. The results show that social representations of sexual violence against women were associated with ideas of suffering, behavioral disturbances, and forced sexual intercourse. The explanations offered for why this type of violence occurs included gender relations, urban violence, and ascription of blame to the victim. It can be concluded that hegemonic patterns of asymmetrical relations persist, even in the discourse of maternity health professionals, who are the point of reference for attending to victims of sexual violence. Incorporating the analytical category of gender into healthcare professional training could make prenatal care an important gateway for the recognition and management of sexual violence against women. PMID- 16470281 TI - [Asthma-related mortality, Brazil, 2000: a study using multiple causes of death]. AB - Asthma is underestimated in mortality statistics that only look at the underlying cause of death. We studied deaths of residents in the Brazilian states of Ceara, Pernambuco, Bahia, Minas Gerais, Rio de Janeiro, Sao Paulo, and Rio Grande do Sul, as well as total deaths in Brazil, where asthma was recorded in any line or part of the death certificate. Causes of death were processed using the Multiple Cause Tabulator. In Brazil, the death rate increased about 50% when all mentions of asthma in death certificates were considered. This increase differed between 36% in Ceara and 76% in Rio Grande do Sul. The highest rates occurred among women and the elderly. The majority of deaths occurred in hospitals, except in Ceara, where the majority occurred at home. Diseases of the circulatory and respiratory systems, emphasizing chronic obstructive pulmonary diseases, were found to be underlying and associated (nonunderlying) causes in deaths related to asthma. Mortality with asthma as the underlying cause was greater in Brazil than in Australia, England and Wales, Scotland, Sweden, and the United States. PMID- 16470282 TI - [Adolescents as research subjects and free informed consent: knowledge and opinion of researchers and adolescents]. AB - This article presents the results of a study that evaluated the knowledge and opinions of researchers and adolescents that served as their research subjects on the legal norms that regulate the participation of the latter as research subjects, the capacity of adolescents to make autonomous decisions regarding participation, and the adolescent experience after agreeing to take part in a study. This was a qualitative study with a convenience sample, the size of which was defined by the criteria of informational redundancy. Interviews were conducted with three researchers who had used adolescents as research subjects and nine of these subjects. This number of interviews was sufficient to reach informational redundancy. Data was collected through recorded semi-structured interviews, with open questions. All the researchers were familiar with some legal document related to the participation of adolescents as subjects of research. On the other hand, the adolescents were surprised because they were not aware of the existence of such documents. However, they considered them necessary for their own protection. In general, researchers and adolescents believe that adolescents have the capacity to decide autonomously to participate as research subjects. The adolescents affirmed that they had decided to volunteer conscientiously. PMID- 16470283 TI - [Association between noise-induced hearing loss and tinnitus]. AB - The objective of this study was to verify an association between noise-induced hearing loss (NIHL) and tinnitus in workers exposed to occupational noise. Workers with a history of work noise exposure who attended two audiology outpatient clinics were interviewed and evaluated. The statistical association between NIHL and tinnitus was evaluated using an adjusted linear regression model, with tinnitus as the dependent variable and NIHL, classified into six levels, as the independent variable, with age and length of noise exposure as covariates. Data were collected from 284 workers between April and October, 2003, in Bauru, Sao Paulo State, Brazil. Tinnitus was observed to increase with the progression of auditory damage, controlled by age and length of noise exposure. These results suggest that investment in hearing conservation programs is needed, especially for source noise emission control and for intervention in hearing loss progression that is caused by noise exposure, in order to maintain auditory health and reduce the associated symptoms. PMID- 16470284 TI - [Factors associated with breastfeeding duration in three cities in the region of Alto Jequitinhonha, Minas Gerais, Brazil]. AB - The objective of this study was to identify and analyze variables associated with shorter duration of breastfeeding in the Alto Jequitinhonha region, Minas Gerais, Brazil. This was a cross-sectional study, using retrospective data, in three cities of the region. All children up to 24 months of age were considered in the study, with a total sample of 450 children. Statistical analyses employed the Kaplan-Meier method and Cox regression model. Three variables were found to be associated with breastfeeding duration. Risk of premature interruption was 1.59 [1.08;2.36] times higher if the father had completed the second grade or greater in school, 1.52 [1.00;2.34] times higher if the father did not live with the child, and 3.07 [2.17;4.34] times higher if child had used a pacifier. Thus, greater education of fathers, use of pacifiers by children, and fathers not living with the children were factors associated with shorter duration of breastfeeding. PMID- 16470285 TI - [Sexual relations and the perception of risk of acquiring STD/AIDS among young adult men in Lima, Peru]. AB - The purpose of this study was to identify men's co-responsibility in the negotiation process of dual protection, according gender, in their first and most recent sexual relationships, and their perception of the risk of STD/AIDS. We surveyed 750 males from 19 to 29 years of age in 2,250 randomly selected households in metropolitan Lima, Peru. The majority mentioned condoms as a contraceptive technique (95%). Their first sexual relationships occurred with a female friend, with a stable partner, or with a known woman. Almost half used some type of protection, principally condoms in order to avoid pregnancy, but their consistent and correct use was low. For 39.5%, the most recent sexual relationship was with a stable partner, a female friend or "known" woman, a wife, or a female co-resident. Dialogue about protection increased between the first and the most recent sexual relationships. The perception of risk of acquiring STD/AIDS was low, and they tended to use protection only with an "unknown partner". PMID- 16470286 TI - [Use of medications by elderly Brazilian retirees: 1--methodology and response rates of a multicenter survey]. AB - This paper reports methodological aspects and response rates of a survey with the main objective of describing the drug utilization profile of retirees of the Institute of National Social Security of Brazil, 60 years of age or older, in three simple random samples: Brazil (countrywide), Belo Horizonte municipality, and Rio de Janeiro municipality. This cross-sectional study employed two approaches: mail (self-report questionnaires) and home interviews (questionnaires completed by interviewers). This strategy allowed evaluation of response agreement between the mail and home interview approaches. In the national sample of 3,000 individuals, 1,025 only responded to the self-report questionnaire. Despite this the low response rate, there was reasonable resemblance between respondents and non-respondents, suggesting that this sample is representative of the target population. In Belo Horizonte and Rio de Janeiro the initial samples of 800 individuals were replicated for both approaches, mail and home interview. The response rates to the mail surveys were 46.8% and 34.4% in Belo Horizonte and Rio de Janeiro, respectively. The response rates to the home interviews were 80.3% and 70.7% in Belo Horizonte and Rio de Janeiro, respectively, after unavoidable attrition. PMID- 16470287 TI - [Prevalence of occupational exposures to potentially infectious materials among dentists and dental assistants]. AB - Dentists and dental assistants work in conditions that favor the occurrence of occupational exposures to potentially infectious materials. The aims of this study are: to determine the prevalence of occupational exposures throughout professional life and in the previous year, to identify the circumstances of exposures, and to verify if there exists a relationship between their occurrence and the use of personal protective equipment. 289 dentists and 104 dental assistants from the city of Florianopolis, Brazil, participated in this study. Data were collected through self-report questionnaires. The prevalence of occupational exposures throughout professional life was higher among dentists (94.5%) than dental assistants (80.8%), while in the previous year it was similar between dentists (39.1%) and assistants (39.4%). However, considering the exposures in the previous year, percutaneous injuries were more frequent among assistants (95.2%) than dentists (60.7%). Consistent use of protective eyewear was statistically associated with less splashes in the eyes in dentists (p = 0,004). Education is recommended to reduce the frequency of occupational exposures in the study population. PMID- 16470288 TI - [Non-medical use of psychoactive medicines among elementary and high school students in Southern Brazil]. AB - The objective of this study is to assess the prevalence of and risk factors for the non-medical use of psychoactive medicines among students at public and private schools of Passo Fundo, Southern Brazil. A cross-sectional study was carried out using a questionnaire administered to 5,057 students from the 5th grade of elementary school to the 3rd year of high school. The questionnaire contained questions about the use of amphetamines, tranquilizers, barbiturates, anticholinergics, opioids, appetite stimulants, and anabolic steroids. Of the sample total, 7.7% had consumed tranquilizers sometime during their lives, 6.4% had used amphetamines, 2.2% had used anabolic steroids, and 1.1% had used barbiturates. Female students reported significantly greater consumption of tranquilizers and amphetamines, while anabolic steroid use was more prevalent among males. The pattern of psychoactive medicine consumption among children and adolescents students is comparable with the pattern among adults. The findings of this research suggest the need to include children and adolescents in media campaigns and other education programs to prevent the non-medical use and abuse of psychoactive medicines. PMID- 16470289 TI - [Perinatal deaths and childbirth healthcare evaluation in maternity hospitals of the Brazilian Unified Health System in Belo Horizonte, Minas Gerais, Brazil, 1999]. AB - This paper analyzes the association between perinatal mortality and factors related to hospital care during labor, considering that healthcare assessment is needed in order to reduce perinatal mortality. A population-based case-control study was conducted with 118 perinatal deaths (cases) and 492 births (controls) that took place in maternity hospitals of the Brazilian Unified Health System (SUS) in Belo Horizonte, Minas Gerais, Brazil. Male sex, prematurity, diseases during pregnancy, low birth weight, newborn diseases, lack of prenatal care, lack of partograph use during labor, and less than one fetus assessment per hour during labor were significantly associated with perinatal deaths. In the multiple regression analysis, lack of partograph use during labor and type of hospital were associated with perinatal deaths. These results indicate inadequate quality of care in maternity hospitals and show that health services structure and health care process are related to perinatal mortality due to preventable causes. PMID- 16470290 TI - [Work conditions and mental health among doctors from Salvador, Bahia, Brazil]. AB - This cross-sectional study investigated the association between work conditions and minor psychological disorders (MPD) in a random sample of 350 doctors from Salvador City, State of Bahia, Brazil. Data were gathered using an individual, self-report questionnaire that incorporated the demand-control model (Job Content Questionnaire) to evaluate psychosocial aspects of the doctors' work and the Self report Questionnaire (SRQ-20) to evaluate their mental health. The study found increased work overload, overtime and extended work shifts, multiple jobs, low income per hour worked, and unstable work arrangements with payment per procedure. The prevalence of MPD was 26.0% and was more strongly associated with aspects of the psychological demands of work than with the doctors' control over it. Doctors with high strain (high demand and low control) jobs showed MPD prevalence of 3.07 (95%IC: 1.38-6.85) times higher than those in low strain (low demand and high control) jobs. PMID- 16470292 TI - [Prevalence of overweight in adolescents: a population-based study in a southern Brazilian city]. AB - The prevalence of overweight and its association with demographic, socioeconomic, behavioral, and biological variables were evaluated. The sample included 810 adolescents (10-19 years of age) living in the urban area of Pelotas, a southern Brazilian city. Overweight was defined as a body mass index > or = the 85th percentile, according to sex and age, and compared to the First National Health and Nutrition Examination Survey, following the WHO recommendation for adolescents. A clustering sampling strategy was used, and both the crude and adjusted analyses (Poisson Regression) took this strategy into account. The prevalence of overweight was 19.3% (95%CI: 16.6-22.0) and there was no difference between the sexes. The following groups presented a greater probability of being overweight: those classified in the wealthiest socioeconomic groups, those who had dieted to lose weight within the previous 3 months, those who watch 4 or more hours of television per day, and those who have less than 3 regular meals per day. After stratification by gender, high socioeconomic level was associated with greatest risk of overweight among boys. Dieting to lose weight during the previous 3 months, 4 or more hours of television viewing per day, and less than 3 formal meals per day were risk factors for overweight among girls. PMID- 16470291 TI - [A comparative analysis of the ambulatory care production register in oncology in the Brazilian Unified Health System]. AB - The response of healthcare services to new information systems depends upon, among other factors, their local organization. This study aimed at comparing the chemotherapy and radiotherapy production registers for the cities Rio de Janeiro and Belo Horizonte, Brazil, in the Ambulatory Information System of the Unified Brazilian Health System (in Portuguese, SUS) between November/1998 and February/2003. Both cities showed a pattern of increase for the chemotherapy production series, while the radiotherapy series showed such increase only for Rio de Janeiro. Comparing Box-Jenkins models for each series, Rio de Janeiro showed a delay of three months in recording production, due to public services. In Belo Horizonte, the delay was shorter than in Rio de Janeiro and there was no difference between public and non-public services. The observed differences between the cities seem to be associated with the fact that Belo Horizonte already had a mechanism to control and evaluate production before the introduction of the system in 1998. PMID- 16470293 TI - [Lead poisoning and child health: integrated efforts to combat this problem]. AB - An epidemiological survey was carried out by technicians of the State Health Secretary and the Municipal Health Secretary of Bauru, Sao Paulo, Brazil, due to excessive atmospheric lead emissions caused by a battery manufacturer. This survey included 853 children from 0 to 12 years old, in a 1,000-meter area from the polluting source, in Bauru (2002). The blood lead levels of children in the exposed group were higher than those in the control group (p < 0.05). 314 children were found to have dosages equal or superior to 10 microg/dl, the limit stipulated by Centers for Disease Control and Prevention. Public services, universities, and volunteers developed some activities aiming at child diagnosis and treatment. The Municipal Health Secretary coordinated remediation initiatives such as: scraping the superficial surface of streets, internal aspiration of houses with professional equipment, and washing and sealing tanks. Through this work, the Lead Poisoning Study and Research Group (GEPICCB) shares an integrated, interdisciplinary, and interinstitutional action proposal. PMID- 16470294 TI - [Prenatal care in an university hospital: evaluating the process]. AB - The objective of this study was to evaluate the prenatal care process referring to the Prenatal Assistance to Low Risk Pregnant Women Program of the University Hospital of Maringa, Parana State, Brazil. It was made a selection of quality criteria which evaluate the precocity in the process and the adequacy in the number of consultation during prenatal period, measured by the Humanization Program of Prenatal and Birth of the Health Ministry and the adequacy on the use of the prenatal care. The study showed that 44.5% of pregnant women started late the prenatal care suggest of the low offer and search to better quality care, which is manifested by spontaneous transference of pregnant women to other services to the program. The prenatal consultations were confirmed and the results showed a mean of 9.8 consultations per pregnant woman--higher than the recommended national parameters. The indicators used and developed to the evaluation of process still present some obstacles to the organizational access and need to define strategies which guarantee this fundamental policy of the Brazilian Unified Health System. PMID- 16470295 TI - [Living conditions and infant mortality in the State of Parana, Brazil, 1997/2001]. AB - This objective of this paper was to verify the relations between the living and infrastructure conditions of municipalities in the State of Parana, Brazil, and infant mortality rate and its components. An ecological study was carried out having the 399 municipalities of the State being classified into five clusters of living conditions. Infant, neonatal and postneonatal rates were calculated for the period 1997/2001. The mean infant, neonatal and postneonatal mortality rates in the State were, respectively, 19.3, 12.0 and 7.3 per 1,000 live births. Postneonatal mortality rates ranged from 8.7 (cluster of worst living conditions) to 6.3 per 1,000 live births (in the cluster classified as having the best living conditions). Regarding the neonatal component, clusters classified as holding the best and the worst living conditions presented the lowest values: 11.8 and 11.4 per 1,000 live births, respectively. We conclude that the postneonatal component still adequately discriminate living conditions in the State of Parana. However, there is a need of complementary studies that aim to clarify the causes of similar neonatal mortality rates observed in the richest and in the poorest cluster of municipalities. PMID- 16470298 TI - [Educators, advisors, therapists? Youth, sexuality, and social intervention]. AB - Sexual and reproductive health programs directed at youth have a recent history, related to international politics, especially the International Conference on Population and Development in Cairo, which in 1994 recommended specific services for adolescents and proposed to reduce substantially the pregnancies in this social group. This effort is an opportunity to consider young people as citizens with rights, or also as new clients and health consumers. In this article, we analyze how this conflict is addressed in the Sexual and reproductive health programs, and the limits and range of the mediating role played by civil employees who carry out health interventions, specially when they enter into relationships with other agents of socialization of adolescents (fathers, mothers, educators, etc.), but also with the public health system. In this field we identified the predominant perspectives of social intervention that seeks to direct and structure the possible field of action of youth in sexual matters. These perspectives emphasize individual responsibility in relation to self care and articulate very well with the effective model of health, based on market rationality. PMID- 16470297 TI - [Juvenile trajectories and social interventions: repercussions for social practices and health (Rio de Janeiro, Brazil)]. AB - This article analyzes the perceptions of a group of youth from the lower economic levels of Rio de Janeiro, Brazil, about the repercussions of social programs on their affective/sexual and educational/professional trajectories. From a comparative perspective, we examined the perspectives, practices, and conditions of life of 42 boys and girls, between 18 and 24 years of age, who did and did not participate in social projects directed at professionalizing and promoting the population. In particular, we discuss the impacts of interventions on concepts of health and AIDS, on indicators of social mobility and professionalization, and on the formation of social networks, including the dimension of gender. We sought to address the effects of civic organizations on the low-income youth population, in the context of socioeconomic crisis and of few social policy advances. We emphasize that in spite of the limitations of such interventions in the greater social context, the experience of social support promoted by the projects contributes to the expansion of life perspectives and the symbolic capital of this social group, with positive impacts for the health field. PMID- 16470299 TI - [Sexual behaviors and incidence of sexual and reproductive health programs among poor secondary students and public university students in Cali, Colombia]. AB - In order to qualitatively evaluate sexual behaviors or practices and the incidence of sexual and reproductive health programs in adolescent and postadolescent users educated in Cali, Colombia, two quota samples were carried out, one in schools in poor neighborhoods of the city and one at Valle University (UNIVALLE). In addition, a limited number of twelve open interviews was made among two different student population groups, the schools and the university. The study addresses the sexual behaviors of adolescents and postadolescents, contraceptive methods and prevention of STDs and HIV/AIDS, and the uses of sexual and reproductive health programs by both populations, controlling for social class (socioeconomic stratum), gender, and race. Two sexual and reproductive health programs were taken as reference: that of PROFAMILIA (Pro-Wellbeing of the Colombian Family Association), and the sexual and reproductive health program of the University Medical Service of the UNIVALLE. Results show how the behaviors and responses to sexual and reproductive health programs are mediated by multiple sociological categories that orient the analysis. PMID- 16470300 TI - [Occupational violence problems in an emergency hospital in Londrina, Parana, Brazil]. AB - Workplace violence is currently a worrying factor at many hospitals. The objectives of this study were to characterize the occupational violence problems detected by health workers in an emergency hospital. The study was exploratory and transversal, with a quantitative data approach. It took place at the emergency ward of a general hospital in Londrina, Parana. The study population included 33 people from the nursing team and 14 medical doctors. Data were collected through interviews with health workers and staff managers, as well as assessment of violence records from the last seven years. 100% of nurses, 88.9% of technicians, 88.2% of nurse assistants, and 85.7 % of doctors said that they had been victims of workplace violence, although not documented. The types of violence suffered included verbal assault (95.2%) and moral and sexual battery (33.3%). Preventive measures to reduce occupational violence should be applied. PMID- 16470301 TI - [Mother-child health and nutrition of Kaiowa and Guarani indigenous children, Caarapo Reserve, Mato Grosso do Sul, Brazil]. AB - The objective of this study was to evaluate the nutritional status of indigenous children and to determine the conditions of mother-child health. A cross sectional study was performed with a sample of 137 children from 0 to 59 months of age from Kaiowa and Guarani indigenous communities, Caarapo Reserve, Mato Grosso do Sul, Brazil. Interviews were carried out using a questionnaire covering the conditions of mother-child health. Nutritional evaluation was performed using anthropometric measurements (weight and height). Results showed that 19.7% of mothers had not undergone prenatal examination and 53.3% had home births. Malnutrition in children was 18.2% and 34.1% for the weight/age and height/age indexes, respectively. The proportion of children with malnutrition, when separated by sex, age and education level of the mother, did not show a significant statistical difference for both indexes. This study documented a high occurrence of infant malnutrition and a worrisome mother-child health situation. PMID- 16470302 TI - Specific structural features of syndecans and heparan sulfate chains are needed for cell signaling. AB - The syndecans, heparan sulfate proteoglycans, are abundant molecules associated with the cell surface and extracellular matrix and consist of a protein core to which heparan sulfate chains are covalently attached. Each of the syndecan core proteins has a short cytoplasmic domain that binds cytosolic regulatory factors. The syndecans also contain highly conserved transmembrane domains and extracellular domains for which important activities are becoming known. These protein domains locate the syndecan on cell surface sites during development and tumor formation where they interact with other receptors to regulate signaling and cytoskeletal organization. The functions of cell surface heparan sulfate proteoglycan have been centered on the role of heparan sulfate chains, located on the outer side of the cell surface, in the binding of a wide array of ligands, including extracellular matrix proteins and soluble growth factors. More recently, the core proteins of the syndecan family transmembrane proteoglycans have also been shown to be involved in cell signaling through interaction with integrins and tyrosine kinase receptors. PMID- 16470303 TI - Pim-1 kinase inhibits the activation of reporter gene expression in Elk-1 and c Fos reporting systems but not the endogenous gene expression: an artifact of the reporter gene assay by transient co-transfection. AB - We have studied the molecular mechanism and signal transduction of pim-1, an oncogene encoding a serine-threonine kinase. This is a true oncogene which prolongs survival and inhibits apoptosis of hematopoietic cells. In order to determine whether the effects of Pim-1 occur by regulation of the mitogen activated protein kinase pathway, we used a transcriptional reporter assay by transient co-transfection as a screening method. In this study, we found that Pim 1 inhibited the Elk-1 and NFkappaB transcriptional activities induced by activation of the mitogen-activated protein kinase cascade in reporter gene assays. However, Western blots showed that the induction of Elk-1-regulated expression of endogenous c-Fos was not affected by Pim-1. The phosphorylation and activation of neither Erk1/2 nor Elk-1 was influenced by Pim-1. Also, in the gel shift assay, the pattern of endogenous NFkappaB binding to its probe was not changed in any manner by Pim-1. These data indicate that Pim-1 does not regulate the activation of Erk1/2, Elk-1 or NFkappaB. These contrasting results suggest a pitfall of the transient co-transfection reporter assay in analyzing the regulation of transcription factors outside of the chromosome context. It ensures that results from reporter gene expression assay should be verified by study of endogenous gene expression. PMID- 16470304 TI - Infection with human papillomaviruses of sexual partners of women having cervical intraepithelial neoplasia. AB - Epidemiological studies show that human papillomaviruses (HPV) are strongly related to cervical cancer and cervical intraepithelial neoplasias (CIN). Unlike the case for women, there are no consistent data on the natural history of HPV in the male population even though these viruses are prevalent in males. We carried out a prospective study to assess the prevalence of HPV in males as well as the factors that determine such infections in 99 male sexual partners of women with CIN. The genitalia of the males were physically examined and subjected to peniscopy for the collection of scrapings which were subjected to the polymerase chain reaction and restriction fragment length polymorphism to detect HPV. Of the 99 males sampled, 54 (54.5%) were positive for HPV DNA, 24% of whom presented normal peniscopy, 28% presented evident clinical lesions and 48% isolated lesions consistent with subclinical infection. In the HPV-negative group, 53% showed normal peniscopy, 4% presented evident clinical lesions and 42% isolated lesions consistent with subclinical infection. The study detected a statistically significant association (P < 0.02, Pearson chi-square test) between HPV infection and both the mean number of sexual partners which a male had during his life and the mean number of sexual partners in the year prior to testing. Viral types 6 and 11 were most frequently encountered. The study shows that infection with HPV was frequent in male sexual partners of women with CIN. PMID- 16470305 TI - Liver mitochondrial dysfunction and oxidative stress in the pathogenesis of experimental nonalcoholic fatty liver disease. AB - Oxidative stress and hepatic mitochondria play a role in the pathogenesis of nonalcoholic fatty liver disease. The aim of the present study was to evaluate the role of hepatic mitochondrial dysfunction and oxidative stress in the pathogenesis of the disease. Fatty liver was induced in Wistar rats with a choline-deficient diet (CD; N = 7) or a high-fat diet enriched with PUFAs-omega-3 (H; N = 7) for 4 weeks. The control group (N = 7) was fed a standard diet. Liver mitochondrial oxidation and phosphorylation were measured polarographically and oxidative stress was estimated on the basis of malondialdehyde and glutathione concentrations. Moderate macrovacuolar liver steatosis was observed in the CD group and mild liver steatosis was observed in the periportal area in the H group. There was an increase in the oxygen consumption rate by liver mitochondria in respiratory state 4 (S4) and a decrease in respiratory control rate (RCR) in the CD group (S4: 32.70 +/- 3.35; RCR: 2.55 +/- 0.15 ng atoms of O2 min-1 mg protein-1) when compared to the H and control groups (S4: 23.09 +/- 1.53, 17.04 +/- 2.03, RCR: 3.15 +/- 0.15, 3.68 +/- 0.15 ng atoms of O2 min-1 mg protein-1, respectively), P < 0.05. Hepatic lipoperoxide concentrations were significantly increased and the concentration of reduced glutathione was significantly reduced in the CD group. A choline-deficient diet causes moderate steatosis with disruption of liver mitochondrial function and increased oxidative stress. These data suggest that lipid peroxidation products can impair the flow of electrons along the respiratory chain, causing overreduction of respiratory chain components and enhanced mitochondrial reactive oxygen species. These findings are important in the pathogenesis of nonalcoholic fatty liver disease. PMID- 16470306 TI - CYP2A6 and CYP2E1 polymorphisms in a Brazilian population living in Rio de Janeiro. AB - Cytochrome P450 (CYP) is a superfamily of enzymes involved in the metabolism of endogenous compounds and xenobiotics. CYP2A6 catalyzes the oxidation of nicotine and the activation of carcinogens such as aflatoxin B1 and nitrosamines. CYP2E1 metabolizes ethanol and other low-molecular weight compounds and can also activate nitrosamines. The CYP2A6 and CYP2E1 genes are polymorphic, altering their catalytic activities and susceptibility to cancer and other diseases. A number of polymorphisms described are ethnic-dependent. In the present study, we determined the genotype and allele frequencies of the main CYP2A6 and CYP2E1 polymorphisms in a group of 289 volunteers recruited at the Central Laboratory of Hospital Universitario Pedro Ernesto. They had been residing in the city of Rio de Janeiro for at least 6 months and were divided into two groups according to skin color (white and non-white). The alleles were determined by allele specific PCR (CYP2A6) or by PCR-RFLP (CYP2E1). The frequencies of the CYP2A6*1B and CYP2A6*2 alleles were 0.29 and 0.02 for white individuals and 0.24 and 0.01 for non-white individuals, respectively. The CYP2A6*5 allele was not found in the population studied. Regarding the CYP2E1*5B allele, we found a frequency of 0.07 in white individuals, which was statistically different (P < 0.05) from that present in non-white individuals (0.03). CYP2E1*6 allele frequency was the same (0.08) in both groups. The frequencies of CYP2A6*1B, CYP2A6*2 and CYP2E1*6 alleles in Brazilians are similar to those found in Caucasians and African Americans, but the frequency of the CYP2E1*5B allele is higher in Brazilians. PMID- 16470307 TI - Lycopene and beta-carotene protect in vivo iron-induced oxidative stress damage in rat prostate. AB - It has been suggested that iron overload may be carcinogenic. In the present study, we evaluated the effect of plasma and prostate carotenoid concentration on oxidative DNA damage in 12-week-old Wistar rats treated with intraperitoneal (ip) ferric nitrilotriacetate (Fe-NTA) (10 mg Fe/kg). Plasma beta-carotene and lycopene concentrations were measured as a function of time after ip injection of carotenoids (10 mg kg(-1) day(-1) beta-carotene or lycopene) in rats. The highest total plasma concentration was reached 3 and 6 h after ip injection of lycopene or beta-carotene, respectively. After 5 days of carotenoid treatment, lycopene and beta-carotene were present in the 0.10-0.51 nmol/g wet tissue range in the prostate. Using a sensitive method to detected 8-oxo-7,8-dihydro-2' deoxyguanosine (8-oxodGuo) by HPLC/EC, the level of 8-oxodGuo in rat prostate DNA was significantly higher (6.3 +/- 0.6 residues/10(6) dGuo) 3 h after Fe-NTA injection compared with control rats (1.7 +/- 0.3 residues/10(6) dGuo). Rats supplemented with lycopene or beta-carotene for 5 days prior to Fe-NTA treatment showed a reduction of about 70% in 8-oxodGuo levels to almost control levels. Compared with control rats, the prostate of Fe-NTA-treated animals showed a 78% increase in malondialdehyde accumulation. Lycopene or beta-carotene pre-treatment almost completely prevented lipid damage. Epidemiological studies have suggested a lower risk of prostate cancer in men reporting a higher consumption of tomato products. However, before associating this effect with tomato sauce constituents, more information is required. The results described here may contribute to the understanding of the protective effects of carotenoids against iron-induced oxidative stress. PMID- 16470308 TI - High expression of human carboxypeptidase M in Pichia pastoris: purification and partial characterization. AB - Carboxypeptidase M (CPM) is an extracellular glycosylphosphatidyl-inositol anchored membrane glycoprotein, which removes the C-terminal basic residues, lysine and arginine, from peptides and proteins at neutral pH. CPM plays an important role in the control of peptide hormones and growth factor activity on the cell surface. The present study was carried out to clone and express human CPM in the yeast Pichia pastoris in order to evaluate the importance of this enzyme in physiological and pathological processes. The cDNA for the enzyme was amplified from total placental RNA by RT-PCR and cloned in the vector pPIC9, which uses the methanol oxidase promoter and drives the expression of high levels of heterologous proteins in P. pastoris. The cpm gene, after cloning and transfection, was integrated into the yeast genome, which produced the active protein. The recombinant protein was secreted into the medium and the enzymatic activity was measured using the fluorescent substrate dansyl-Ala-Arg. The enzyme was purified by a two-step protocol including gel filtration and ion-exchange chromatography, resulting in a 1753-fold purified active protein (16474 RFU mg protein(-1) min(-1)). This purification protocol permitted us to obtain 410 mg of the purified protein per liter of fermentation medium. SDS-PAGE showed that recombinant CPM migrated as a single band with a molecular mass similar to that of native placental enzyme (62 kDa), suggesting that the expression of a glycosylated protein had occurred. These results demonstrate for the first time the establishment of a method using P. pastoris to express human CPM necessary to the development of specific antibodies and antagonists, and the analysis of the involvement of this peptidase in different physiological and pathological processes. PMID- 16470309 TI - Prevalence of the A1555G (12S rRNA) and tRNASer(UCN) mitochondrial mutations in hearing-impaired Brazilian patients. AB - Mitochondrial mutations are responsible for at least 1% of the cases of hereditary deafness, but the contribution of each mutation has not yet been defined in African-derived or native American genetic backgrounds. A total of 203 unselected hearing-impaired patients were screened for the presence of the mitochondrial mutation A1555G in the 12S rRNA gene and mutations in the tRNASer(UCN) gene in order to assess their frequency in the ethnically admixed Brazilian population. We found four individuals with A1555G mutation (2%), which is a frequency similar to those reported for European-derived populations in unselected samples. On the other hand, complete sequencing of the tRNASer(UCN) did not reveal reported pathogenic substitutions, namely A7445G, 7472insC, T7510C, or T7511C. Instead, other rare substitutions were found such as T1291C, A7569G, and G7444A. To evaluate the significance of these findings, 110 "European Brazilians" and 190 "African-Brazilians" unrelated hearing controls were screened. The T1291C, A7569G and G7444A substitutions were each found in about 1% (2/190) of individuals of African ancestry, suggesting that they are probably polymorphic. Our results indicate that screening for the A1555G mutation is recommended among all Brazilian deaf patients, while testing for mutations in the tRNASer(UCN) gene should be considered only when other frequent deafness-causing mutations have been excluded or in the presence of a maternal transmission pattern. PMID- 16470310 TI - Effects of gamma-radiation on cell growth, cycle arrest, death, and superoxide dismutase expression by DU 145 human prostate cancer cells. AB - Gamma-irradiation (gamma-IR) is extensively used in the treatment of hormone resistant prostate carcinoma. The objective of the present study was to investigate the effects of 60Co gamma-IR on the growth, cell cycle arrest and cell death of the human prostate cancer cell line DU 145. The viability of DU 145 cells was measured by the Trypan blue exclusion assay and the 3(4,5 dimethylthiazol-2-yl)-2,5,diphenyltetrazolium bromide test. Bromodeoxyuridine incorporation was used for the determination of cell proliferation. Cell cycle arrest and cell death were analyzed by flow cytometry. Superoxide dismutase (SOD), specifically CuZnSOD and MnSOD protein expression, after 10 Gy gamma-IR, was determined by Western immunoblotting analysis. Gamma-IR treatment had a significant (P < 0.001) antiproliferative and cytotoxic effect on DU 145 cells. Both effects were time and dose dependent. Also, the dose of gamma-IR which inhibited DNA synthesis and cell proliferation by 50% was 9.7 Gy. Furthermore, gamma-IR induced cell cycle arrest in the G2/M phase and the percentage of cells in the G2/M phase was increased from 15% (control) to 49% (IR cells), with a nonsignificant induction of apoptosis. Treatment with 10 Gy gamma-IR for 24, 48, and 72 h stimulated CuZnSOD and MnSOD protein expression in a time-dependent manner, approximately by 3- to 3.5-fold. These data suggest that CuZnSOD and MnSOD enzymes may play an important role in the gamma-IR-induced changes in DU 145 cell growth, cell cycle arrest and cell death. PMID- 16470311 TI - The P48T germline mutation and polymorphism in the CDKN2A gene of patients with melanoma. AB - CDKN2A has been implicated as a melanoma susceptibility gene in some kindreds with a family history of this disease. Mutations in CDKN2A may produce an imbalance between functional p16ink4a and cyclin D causing abnormal cell growth. We searched for germline mutations in this gene in 22 patients with clinical criteria of hereditary cancer (early onset, presence of multiple primary melanoma or 1 or more first- or second-degree relatives affected) by secondary structural content prediction, a mutation scanning method that relies on the propensity for single-strand DNA to take on a three-dimensional structure that is highly sequence dependent, and sequencing the samples with alterations in the electrophoretic mobility. The prevalence of CDKN2A mutation in our study was 4.5% (1/22) and there was a correlation between family history and probability of mutation detection. We found the P48T mutation in 1 patient with 2 melanoma affected relatives. The patient descends from Italian families and this mutation has been reported previously only in Italian families in two independent studies. This leads us to suggest the presence of a mutational "hotspot" within this gene or a founder mutation. We also detected a high prevalence (59.1%) of polymorphisms, mainly alleles 500 C/G (7/31.8%) or 540 C/T (6/27.3%), in the 3' untranslated region of exon 3. This result reinforces the idea that these rare polymorphic alleles have been significantly associated with the risk of developing melanoma. PMID- 16470312 TI - Cyclosporin-A does not affect skeletal muscle mass during disuse and recovery. AB - Cyclosporin-A (CsA) is an immunosuppressive drug that acts as an inhibitor of calcineurin, a calcium phosphatase that has been suggested to play a role in skeletal muscle hypertrophy. The aim of the present study was to determine the effect of CsA administration (25 mg kg(-1) day(-1)) on skeletal muscle mass and phenotype during disuse and recovery. Male Wistar rats received vehicle (N = 8) or CsA (N = 8) during hind limb immobilization (N = 8) and recovery (N = 8). Muscle weight (dry/wet) and cross-sectional area were evaluated to verify the effect of CsA treatment on muscle mass. Muscle phenotype was assessed by histochemistry of myosin ATPase. CsA administration during immobilization and recovery did not change muscle/body weight ratio in the soleus (SOL) or plantaris (PL). Regarding muscle phenotype, we observed a consistent slow-to-fast shift in all experimental groups (immobilized only, receiving CsA only, and immobilized receiving CsA) as compared to control in both SOL and PL (P < 0.05). During recovery, no difference was observed in SOL or PL fiber type composition between the experimental recovered group and recovered group receiving CsA compared to their respective controls. Considering the muscle/body weight ratio, CsA administration does not maximize muscle mass loss induced by immobilization. Our results also indicate that CsA fails to block skeletal muscle regrowth after disuse. The present data suggest that calcineurin inhibition by CsA modulates muscle phenotype rather than muscle mass. PMID- 16470313 TI - The role of IFN-gamma and IL-4 in gastric mucosa inflammation associated with Helicobacter heilmannii type 1 infection. AB - Although Helicobacter heilmannii infection is less common than H. pylori infection in humans, it is considered to be of medical importance because of its association with gastritis, gastric ulcer, carcinoma, and mucosa-associated lymphoid tissue lymphoma of the stomach. However, there have been no studies evaluating the role of the Th cell response in H. heilmannii gastric infection. We evaluated the participation of pro-inflammatory and anti-inflammatory cytokines, IFN-gamma and IL-4, in H. heilmannii gastric infection in genetically IFN-gamma- or IL-4-deficient mice. The serum IFN-gamma and IL-4 concentrations were determined by ELISA. The gastric polymorphonuclear infiltrate was higher (P = 0.007) in H. heilmannii-positive than in H. heilmannii-negative wild-type (WT) C57BL/6 mice, whereas no significant inflammation was demonstrable in the stomach of H. heilmannii-positive IFN-gamma(-/-) C57BL/6 mice. The degree of gastric inflammatory cells, especially in oxyntic mucosa, was also higher (P = 0.007) in infected IL-4(-/-) than in WT BALB/c mice. Serum IFN-gamma levels were significantly higher in IL-4(-/-) than in WT BALB/c mice, independently of H. heilmannii-positive or -negative status. Although no difference in serum IFN gamma levels was seen between H. heilmannii-positive (11.3 +/- 3.07 pg/mL, mean +/- SD) and -negative (11.07 +/- 3.5 pg/mL) WT BALB/c mice, in the group of IL-4( /-) animals, the serum concentration of IFN-gamma was significantly higher in the infected ones (38.16 +/- 10.5 pg/mL, P = 0.04). In contrast, serum IL-4 levels were significantly decreased in H. heilmannii-positive (N = 10) WT BALB/c animals compared to the negative (N = 10) animals. In conclusion, H. heilmannii infection induces a predominantly Th1 immune response, with IFN-gamma playing a central role in gastric inflammation. PMID- 16470315 TI - Activation of neural cholecystokinin-1 receptors induces relaxation of the isolated rat duodenum which is reduced by nitric oxide synthase inhibitors. AB - Cholecystokinin (CCK) influences gastrointestinal motility, by acting on central and peripheral receptors. The aim of the present study was to determine whether CCK has any effect on isolated duodenum longitudinal muscle activity and to characterize the mechanisms involved. Isolated segments of the rat proximal duodenum were mounted for the recording of isometric contractions of longitudinal muscle in the presence of atropine and guanethidine. CCK-8S (EC50: 39; 95% CI: 4.1-152 nM) and cerulein (EC50: 58; 95% CI: 18-281 nM) induced a concentration dependent and tetrodotoxin-sensitive relaxation. Nomeganitro-L-arginine (L-NOARG) reduced CCK-8S- and cerulein-induced relaxation (IC50: 5.2; 95% CI: 2.5-18 microM) in a concentration-dependent manner. The magnitude of 300 nM CCK-8S induced relaxation was reduced by 100 microM L-NOARG from 73 +/- 5.1 to 19 +/- 3.5% in an L-arginine but not D-arginine preventable manner. The CCK-1 receptor antagonists proglumide, lorglumide and devazepide, but not the CCK-2 receptor antagonist L-365,260, antagonized CCK-8S-induced relaxation in a concentration dependent manner. These findings suggest that CCK-8S and cerulein activate intrinsic nitrergic nerves acting on CCK-1 receptors in order to cause relaxation of the rat duodenum longitudinal muscle. PMID- 16470314 TI - Effect of Erythrina velutina and Erythrina mulungu in rats submitted to animal models of anxiety and depression. AB - Erythrina velutina (EV) and Erythrina mulungu (EM), popularly used in Brazil as tranquilizing agents, were studied. The effects of acute and chronic oral treatment with a water:alcohol extract of EV (7:3, plant grounded stem bark; acute = 100, 200, 400 mg/kg; chronic = 50, 100, 200 mg/kg) were evaluated in rats (N = 11-12) submitted to the elevated T-maze (for avoidance and escape measurements) model of anxiety. This model was selected for its presumed capacity to elicit specific subtypes of anxiety disorders recognized in clinical practice: avoidance has been related to generalized anxiety and escape to panic. Additionally, animals were treated with the same doses of EV and EM (water:alcohol 7:3, inflorescence extract) and submitted to the forced swim test for the evaluation of antidepressant activity (N = 7-10). Both treatment regimens with EV impaired elevated T-maze avoidance latencies, without altering escape, in a way similar to the reference drug diazepam (avoidance 1, mean +/- SEM, acute study: 131.1 +/- 45.5 (control), 9.0 +/- 3.3 (diazepam), 12.7 +/- 2.9 (200 mg/kg), 28.8 +/- 15.3 (400 mg/kg); chronic study: 131.7 +/- 46.9 (control), 35.8 +/- 29.7 (diazepam), 24.4 +/- 10.4 (50 mg/kg), 29.7 +/- 11.5 (200 mg/kg)). Neither EV nor EM altered measurements performed in the forced swim test, in contrast to the reference drug imipramine that significantly decreased immobility time after chronic treatment. These results were not due to motor alterations since no significant effects were detected in an open field. These observations suggest that EV exerts anxiolytic-like effects on a specific subset of defensive behaviors which have been associated with generalized anxiety disorder. PMID- 16470316 TI - Effect of topical dorzolamide on rabbit central corneal thickness. AB - Our objective was to study the effect of dorzolamide on corneal hydration in an 18-week controlled experiment using ultrasonic pachymetry. Twenty-eight male rabbits were divided randomly into four groups. The 7 rabbits in each group received eye drops containing either 2% (w/v) dorzolamide or placebo in their right eye, or in their left eye. The 2% dorzolamide rabbits were treated every 8 h. Fellow eyes are defined as eyes which did not receive either dorzolamide or placebo. The study was blind for both the person who applied the drug and the one who performed the pachymetry. The effect of treatments is reported on the basis of the percentage of pachymetric variation compared to the measurement made before drug application. There was no significant difference (P = 0.061) in pachymetric variation between dorzolamide (-4.42 +/- 11.71%) and placebo (2.48 +/ 9.63%). However, there was a significant difference (P = 0.0034) in pachymetric variation between the dorzolamide fellow eyes (-7.56 +/- 10.50%) and the placebo (-4.42 +/- 11.71%). In conclusion, dorzolamide did not increase the corneal thickness in rabbits. PMID- 16470317 TI - Protective effects of phosphodiesterase inhibitors on lung function and remodeling in a murine model of chronic asthma. AB - The aim of the present study was to compare the efficacy of a novel phosphodiesterase 4 and 5 inhibitor, LASSBio596, with that of dexamethasone in a murine model of chronic asthma. Lung mechanics (airway resistance, viscoelastic pressure, and static elastance), histology, and airway and lung parenchyma remodeling (quantitative analysis of collagen and elastic fiber) were analyzed. Thirty-three BALB/c mice were randomly assigned to four groups. In the asthma group (N = 9), mice were immunized with 10 microg ovalbumin (OVA, ip) on 7 alternate days, and after day 40 they were challenged with three intratracheal instillations of 20 microg OVA at 3-day intervals. Control mice (N = 8) received saline under the same protocol. In the dexamethasone (N = 8) and LASSBio596 (N = 8) groups, the animals of the asthma group were treated with 1 mg/kg dexamethasone disodium phosphate (0.1 mL, ip) or 10 mg/kg LASSBio596 dissolved in dimethyl sulfoxide (0.2 mL, ip) 24 h before the first intratracheal instillation of OVA, for 8 days. Airway resistance, viscoelastic pressure and static elastance increased significantly in the asthma group (77, 56, and 76%, respectively) compared to the control group. The asthma group presented more intense alveolar collapse, bronchoconstriction, and eosinophil and neutrophil infiltration than the control group. Both LASSBio596 and dexamethasone inhibited the changes in lung mechanics, tissue cellularity, bronchoconstriction, as well as airway and lung parenchyma remodeling. In conclusion, LASSBio596 at a dose of 10 mg/kg effectively prevented lung mechanical and morphometrical changes and had the potential to block fibroproliferation in a BALB/c mouse model of asthma. PMID- 16470318 TI - Fecal estradiol and progesterone metabolite levels in the three-toed sloth (Bradypus variegatus). AB - The present study was carried out to assess the possibility of measuring fecal steroid hormone metabolites as a noninvasive technique for monitoring reproductive function in the three-toed sloth, Bradypus variegatus. Levels of the estradiol (E2) and progesterone (P4) metabolites were measured by radioimmunoassay in fecal samples collected over 12 weeks from 4 captive female B. variegatus sloths. The validation of the radioimmunoassay for evaluation of fecal steroid metabolites was carried out by collecting 10 blood samples on the same day as defecation. There was a significant direct correlation between the plasma and fecal E2 and P4 levels (P < 0.05, Pearson's test), thereby validating this noninvasive technique for the study of the estrous cycle in these animals. Ovulation was detected in two sloths (SL03 and SL04) whose E2 levels reached 2237.43 and 6713.26 pg/g wet feces weight, respectively, for over four weeks, followed by an increase in P4 metabolites reaching 33.54 and 3242.68 ng/g wet feces weight, respectively. Interestingly, SL04, which presented higher levels of E2 and P4 metabolites, later gave birth to a healthy baby sloth. The results obtained indicate that this is a reliable technique for recording gonadal steroid secretion and thereby reproduction in sloths. PMID- 16470319 TI - The decreased oxygen uptake during progressive exercise in ischemia-induced heart failure is due to reduced cardiac output rate. AB - We tested the hypothesis that the inability to increase cardiac output during exercise would explain the decreased rate of oxygen uptake (VO2) in recent onset, ischemia-induced heart failure rats. Nine normal control rats and 6 rats with ischemic heart failure were studied. Myocardial infarction was induced by coronary ligation. VO2 was measured during a ramp protocol test on a treadmill using a metabolic mask. Cardiac output was measured with a flow probe placed around the ascending aorta. Left ventricular end-diastolic pressure was higher in ischemic heart failure rats compared with normal control rats (17 +/- 0.4 vs 8 +/ 0.8 mmHg, P = 0.0001). Resting cardiac index (CI) tended to be lower in ischemic heart failure rats (P = 0.07). Resting heart rate (HR) and stroke volume index (SVI) did not differ significantly between ischemic heart failure rats and normal control rats. Peak VO2 was lower in ischemic heart failure rats (73.72 +/- 7.37 vs 109.02 +/- 27.87 mL min(-1) kg(-1), P = 0.005). The VO2 and CI responses during exercise were significantly lower in ischemic heart failure rats than in normal control rats. The temporal response of SVI, but not of HR, was significantly lower in ischemic heart failure rats than in normal control rats. Peak CI, HR, and SVI were lower in ischemic heart failure rats. The reduction in VO2 response during incremental exercise in an ischemic model of heart failure is due to the decreased cardiac output response, largely caused by depressed stroke volume kinetics. PMID- 16470320 TI - Effect of unsaturated fatty acids on myocardial performance, metabolism and morphology. AB - Diets rich in saturated fatty acids are one of the most important causes of atherosclerosis in men, and have been replaced with diets rich in unsaturated fatty acids (UFA) for the prevention of this disorder. However, the effect of UFA on myocardial performance, metabolism and morphology has not been completely characterized. The objective of the present investigation was to evaluate the effects of a UFA-rich diet on cardiac muscle function, oxidative stress, and morphology. Sixty-day-old male Wistar rats were fed a control (N = 8) or a UFA rich diet (N = 8) for 60 days. Myocardial performance was studied in isolated papillary muscle by isometric and isotonic contractions under basal conditions after calcium chloride (5.2 mM) and ss-adrenergic stimulation with 1.0 microM isoproterenol. Fragments of the left ventricle free wall were used to study oxidative stress and were analyzed by light microscopy, and the myocardial ultrastructure was examined in left ventricle papillary muscle. After 60 days the UFA-rich diet did not change myocardial function. However, it caused high lipid hydroperoxide (176 +/- 5 vs 158 +/- 5, P < 0.0005) and low catalase (7 +/- 1 vs 9 +/- 1, P < 0.005) and superoxide-dismutase (18 +/- 2 vs 27 +/- 5, P < 0.005) levels, and discrete morphological changes in UFA-rich diet hearts such as lipid deposits and mitochondrial membrane alterations compared to control rats. These data show that a UFA-rich diet caused myocardial oxidative stress and mild structural alterations, but did not change mechanical function. PMID- 16470323 TI - Atherothrombosis in acute coronary syndromes: mechanisms, markers, and mediators of vulnerability. AB - We review the concept of atherothrombosis and the critically important role of inflammation in the development of acute coronary syndromes (ACS). Inflammation is now known to be a major driving force underlying the initiation of coronary plaques, their unstable progression, and eventual disruption, and it also contributes significantly to thrombotic complications that occur in ACS. In addition, we discuss the various local mediators and systemic markers that are involved in the inflammatory process, and review the concepts of 'vulnerable plaque,' 'vulnerable blood,' 'vulnerable myocardium,' and the 'vulnerable patient' who is at increased risk for ACS. PMID- 16470324 TI - New approaches to out-of-hospital cardiac arrest. AB - Out-of-hospital cardiac arrest (OOH-CA) is a leading cause of mortality and the focus of significant research. Recent studies provide new evidence that may change our management of OOH-CA and improve outcomes. The findings of two recently published studies of OOH-CA are reviewed in this article. The first, the Public Access Defibrillation Trial, was a randomized, controlled trial of public access defibrillation in 993 community facilities in the U.S. and Canada . It demonstrated that a community strategy to train laypersons to respond to cardiac arrests significantly increased survival to hospital discharge following OOH-CA in nonresidential community units with community members trained and equipped to provide public access defibrillation, compared to community units with community members trained to provide cardiopulmonary resuscitation (CPR) without any capacity for defibrillation. The second, the European Resuscitation Council Vasopressor during Cardiopulmonary Resuscitation Study, was a randomized, controlled, double-blinded trial that compared vasopressin to epinephrine as the initial pharmacological therapy for 1,219 patients who sustained OOH-CA. The study demonstrated that vasopressin is similar to epinephrine for OOH-CA due to ventricular fibrillation or pulseless electrical activity, and superior to epinephrine for the initial treatment of asystolic arrest; it also demonstrated that the combination of vasopressin and epinephrine is superior to epinephrine alone in the treatment of refractory, out-of-hospital cardiac arrest. Studies on alternative CPR techniques and adjunctive devices for CPR were also reviewed. We conclude that pre-hospital access to defibrillators and the use of vasopressin in the management of asystole hold promise for improving survival for patients with out-of-hospital cardiac arrest. PMID- 16470325 TI - Unstable angina and non-ST-segment myocardial infarction: an evidence-based approach to management. AB - Unstable angina (UA) and non-ST-segment elevation myocardial infarction (NSTEMI) represent two common, closely related acute coronary syndromes with potentially high morbidity and mortality. Integration of information from the history, physical exam, electrocardiogram, and cardiac biomarkers is used to formulate both the diagnosis of UA/NSTEMI and the overall assessment of patient prognosis and risk. Early diagnosis and risk stratification of patients with UA/NSTEMI enable the physician to initiate timely, appropriate treatment. (There is strong clinical evidence supporting the tailoring of specific therapies to the risk profile of the patient.) In recent years, powerful new medical and invasive therapies have been developed. Pharmaceutical agents for UA/NSTEMI may be broadly grouped into one of three categories: anti-ischemic, anti-platelet, and anti thrombotic agents. Standard therapy for UA/NSTEMI has commonly included oxygen, aspirin, nitrates, morphine, beta-blockers and heparin. Potent new anti-platelet agents, including inhibitors of platelet adenosine diphosphate and glycoprotein IIb/IIIa receptors, play important, expanding roles in the management of these syndromes. Low-molecular-weight heparins have been shown to be an effective alternative to unfractionated heparin in their treatment. Major advances in invasive techniques and devices over the last decade include revascularization with percutaneous coronary intervention and drug-eluting intracoronary stents. Strong interest exists in studying the potential benefits and risks associated with an early invasive therapeutic strategy rather than an aggressive medical regimen for patients with UA/NSTEMI. As new treatments are rapidly added to our growing arsenal of management options, clinicians are constantly challenged with incorporating complex new information and guidelines into their practices in a timely fashion. To assist clinicians with this challenge, this article will review the evidence to support the use of current therapeutic options for UA/NSTEMI, with an emphasis on summarizing the most recent clinical guidelines jointly published by the American College of Cardiology and the American Heart Association. PMID- 16470326 TI - Diagnosis and management of ST elevation myocardial infarction: a review of the recent literature and practice guidelines. AB - There is a large volume of literature available to guide the peri-infarct management of ST elevation myocardial infarction (STEMI). Most of this literature focuses on improving the availability and efficacy of reperfusion therapy. The purpose of this article is to review contemporary scientific evidence and guideline recommendations regarding the diagnosis and therapy of STEMI. Studies and epidemiological data were identified using Medline, the Cochrane Database, and an Internet search engine. Medline was searched for landmark and recent publications using the following key words: STEMI, guidelines, epidemiology, reperfusion, fibrinolytics, percutaneous coronary intervention (PCI), facilitated PCI, transfer, delay, clopidogrel, glycoprotein IIb/IIIa, low-molecular-weight heparin (LMWH), beta-blockers, nitrates, and angiotensin-converting enzyme (ACE) inhibitors. The data accessed indicate that urgent reperfusion with either fibrinolytics or percutaneous intervention should be considered for every patient having symptoms of myocardial infarction with ST segment elevation or a bundle branch block. The utility of combined mechanical and pharmacological reperfusion is currently under investigation. Ancillary treatments may utilize clopidogrel, glycoprotein IIb/IIIa inhibitors, or low molecular weight heparin, depending on the primary reperfusion strategy used. Comprehensive clinical practice guidelines incorporate much of the available contemporary evidence, and are important resources for the evidence-based management of STEMI. PMID- 16470327 TI - Atrial fibrillation. AB - In 2000, some 2.3 million Americans were affected by atrial fibrillation, and that number is expected to rise as our population ages. Atrial fibrillation is both a reflection of active physiologic stressors on the body and a marker of future cardiac disease progression. The disorganized atrial activity that characterizes atrial fibrillation affects cardiac function, metabolic demand, and quality of life. However, our understanding of the etiology and treatment of this condition continues to advance with the result of recent large-scale clinical trials. Diabetes, hypertension, congestive heart failure, valvular disease, and myocardial infarction are all risk factors in the development of atrial fibrillation. And the diagnosis confers a five-fold increase in the incidence of stroke. (Patients at increased risk for stroke include those with congestive heart failure, hypertension, age greater than 75, diabetes, and previous stroke.) Anticoagulation is a critical action in most cases of atrial fibrillation, as data show a 68% relative risk reduction of stroke when patients are treated with warfarin. Prior to recent trials, achieving sinus rhythm was thought to invariably improve symptoms, cardiac function, and mortality. The adverse effects of antiarrhythmic medications are now being recognized, and treatment strategies emphasizing ventricular rate control have been recommended in recent clinical practice guidelines. This shift in thinking is influencing both outpatient and emergency department management. Controlling the ventricular rate in atrial fibrillation increases cardiac output, decreases the metabolic demand of the heart, and avoids the potentially dangerous side effects of rhythm-control drugs. Rate-control agents should be selected based on the clinical profile of individual patients. A well-chosen subset of patients may benefit from either chemical or electrical cardioversion; this appears to be a reasonably safe procedure and can be accomplished on an outpatient basis. Understanding causal etiologies, managing risk for stroke (and need for anticoagulation), addressing rate, and assessing the risks of cardioversion are key elements in a comprehensive approach to atrial fibrillation. PMID- 16470328 TI - Treatment of tension-type headache with botox: a review of the literature. AB - Botulinum toxin has been shown to effectively treat several types of neurological disorders. It has recently been evaluated for the treatment of tension-type headaches in patients who are unable to tolerate or cannot benefit from standard therapies. Most of the open design studies seem to present positive results. However, the randomized, double-blind, placebo-controlled studies present contradictory results for the efficacy of botulinum toxin. Based on these data, further controlled trials of botulinum toxin are needed to evaluate its effects on tension headaches and to determine optimal injection sites, doses, and frequency of treatments. PMID- 16470330 TI - Detection of novel Y SNPs provides further insights into Y chromosomal variation in Pakistan. AB - Biallelic polymorphisms on the Y chromosome have been extensively used to study the history, evolution, and migration patterns of world populations. In this study we screened 8.5 kb of Y chromosomal DNA for single nucleotide polymorphisms (SNPs) in a panel of 95 male individuals belonging to different haplogroups. Five novel Y-SNPs (PK1-5) were identified, four in the Pakistani sample and one in an African sample. The ancestral state of each SNP was determined in two chimpanzee samples and a variety of Pakistani ethnic groups. In addition to these novel Y SNPs 77 additional markers on the Y chromosome were analyzed to place the SNPs on the phylogenetic tree of Y chromosomal lineages and to further investigate extant human Y chromosomal variation within Pakistan. BATWING analysis gave an estimate of between 2,500 and 7,300 YBP for population expansion in Pakistan which coincides with the period of the Indus Valley civilizations. PMID- 16470329 TI - Unified method for Bayesian calculation of genetic risk. AB - Bayesian inference has been used for genetic risk calculation. In this traditional method, inheritance events are divided into a number of cases under the inheritance model, and some elements of the inheritance model are usually disregarded. We developed a genetic risk calculation program, GRISK, which contains an improved Bayesian risk calculation algorithm to express the outcome of inheritance events with inheritance vectors, a set of ordered genotypes of founders, and mutation vectors, which represent a new idea for description of mutations in a pedigree. GRISK can calculate genetic risk in a common format that allows users to execute the same operation in every case, whereas the traditional risk calculation method requires construction of a calculation table in which the inheritance events are variously divided in each respective case. In addition, GRISK does not disregard any possible events in inheritance. This program was developed as a Japanese macro for Excel to run on Windows. PMID- 16470331 TI - Sensitivity and utility of parathyroid scintigraphy in patients with primary versus secondary and tertiary hyperparathyroidism. AB - INTRODUCTION: Parathyroid scintigraphy (PS) may be used to localize hyperactive parathyroid glands preoperatively. Performance of PS in the setting of secondary and tertiary hyperparathyroidism (HPT) is not well quantified. The performance of PS in secondary/tertiary HPT versus primary HPT may reflect physiologic as well as radiopharmaceutical kinetic differences between multigland hyperplasia versus adenoma. The aim of this study was to review the performance of PS in secondary/tertiary HPT with a comparison to that for primary HPT. Moreover, we evaluated (1) the sensitivity of PS in detecting enlarged glands, and (2) PS detectability as a function of gland weight. METHODS: We performed a retrospective review of the Mayo Clinic database from 2000 to 2004. We identified 40 patients with secondary or tertiary HPT as well as a matched control group of 40 patients with primary HPT who had had preoperative PS and underwent parathyroid surgery. RESULTS: Parathyroid scintigraphy correctly localized all enlarged glands in 88% of patients in the primary HPT group. PS correctly identified both the number and locations of all hyperplastic glands in only 28% of the secondary/tertiary HPT patients. PS failed to identify one enlarged gland in 23% of the patients and two or more enlarged glands in 40% of the patients. PS correctly detects the largest gland in 88% of the patients with secondary and tertiary HPT. The mean gland weight detectable by PS was 612 +/- 120 mg for primary HPT. In secondary/tertiary HPT, glands detected by PS had a mean weight of 950 +/- 109 mg, whereas the mean weight was 276 +/- 34 mg for undetected glands (P < 0.002). CONCLUSIONS: Parathyroid scintigraphy is a sensitive study for localizing parathyroid glands preoperatively in primary HPT patients. Its sensitivity is low in secondary and tertiary HPT patients. Thus PS has limited value as a preoperative localization study in secondary/tertiary HPT patients. PMID- 16470332 TI - The stroke size of myosins: a reevaluation. AB - In this article results are reviewed from different experimental approaches to determine the size of the power stroke generated by myosin molecules during their ATPase cycle. While data from fiber studies and protein crystallography predict a stroke size of about 10 nm for skeletal muscle myosins, single molecule studies imply a stroke size for these myosins of only about 5 nm. Single molecule studies also showed the stroke size to be proportional to the length of the light chain binding domain, acting like a lever arm. At the same lever arm length, however, the stroke size of smooth muscle myosin II is found about twice as large and a stroke size of about 14 nm was reported for class-I myosins. It was proposed that such different stroke sizes for molecules with same lever arm length result from different extend of converter domain rotation. Only for class-I myosins, however, an about 30 degrees larger rotation of the converter was found so far by protein crystallography. This, however, is far too small to account for the almost 3-fold larger stroke size reported from single molecule studies. In this contribution we discuss some factors that might account for the apparent discrepancies between single molecule studies on the one hand and protein crystallography as well as some fiber studies on the other hand. In addition, we present some modeling to illustrate that the power stroke very likely is underestimated to a large extent in current single molecule approaches. We further show that differences in the stroke size for various classes of myosins reported from single molecule studies might be related to small differences in the probability to execute the power stroke kinetics. We demonstrate that such small changes in power stroke kinetics can seriously affect the extent to which the 'true' power stroke is underestimated by present single molecule approaches. PMID- 16470333 TI - Functional and biophysical analyses of the class XIV Toxoplasma gondii myosin D. AB - The obligate intracellular parasite Toxoplasma gondii uses gliding motility to migrate across the biological barriers of the host and to invade cells. This unique form of locomotion requires an intact actin cytoskeleton and involves at least one motor protein (TgMyoA) that belongs to the class XIV of the myosin superfamily. TgMyoA is anchored in the inner membrane complex and is essential for the gliding motion, host cell invasion and egress of T. gondii tachyzoites. TgMyoD is the smallest T. gondii myosin and is structurally very closely related to TgMyoA. We show here that TgMyoD exhibits similar transient kinetic properties as the fast single-headed TgMyoA. To determine if TgMyoD also contributes to parasite gliding motility, the TgMyoD gene was disrupted by double homologous recombination. In contrast to TgMyoA, TgMyoD gene is dispensable for tachyzoite propagation and motility. Parasites lacking TgMyoD glide normally and their virulence is not compromised in mice. The fact that TgMyoD is predominantly expressed in bradyzoites explains the absence of a phenotype observed with myodko in tachyzoites and does not exclude a role of this motor in gliding that would be restricted to the cyst forming but nevertheless motile stage of the parasite. PMID- 16470334 TI - Restoring force development by titin/connectin and assessment of Ig domain unfolding. AB - Titin/connectin is the main determinant of physiological levels of passive muscle force. This force is generated by the extensible I-band region of the molecule, which is composed of serially-linked immunoglobulin (Ig)-like domains and several unique sequence elements. Here we address the role of titin/connectin in sarcomeres shortened to below the slack length (length attained by an un activated cell in absence of external forces). Such shortened cells develop so called restoring forces that re-extend the cells upon relaxation. The experiments that we present are based on a high throughput method with a rapid solution switching system which allows unattached single cardiac myocytes to be activated (resulting in shortening below the slack length) and then to be rapidly relaxed while their maximal re-lengthening velocity is measured at the sarcomere level (dSL/dtmax), with high-resolution imaging techniques. Experiments were carried out on myocytes that express different isoforms of titin/connectin. We measured the relation between dSL/dtmax and the minimal SL during contraction (SLmin) and determined the slope of this relation as a measure of 'restoring stiffness.' We found that the restoring stiffness correlates with the isoform expression profile with myocytes that express high levels of the stiff isoform (N2B) having the highest restoring stiffness. These results support the notion that titin/connectin is a bi-directional spring that develops passive force when stretched above the slack length and restoring force when shortened to below this length. We also discuss in detail the mechanisms that underlie titin/connectin's restoring force development and focus on whether or not unfolding of Ig domains plays a role. PMID- 16470335 TI - Partial sequence of connectin-like 1200K-protein in obliquely striated muscle of a polychaete (Annelida): evidence for structural diversity from vertebrate and invertebrate connectins. AB - Vertebrate striated muscle contains the giant elastic protein connectin that maintains the position of the A-band at the center of the sarcomere during repeated muscular contraction and relaxation. Connectin-like molecules may perform conserved functions in vertebrate and invertebrate striated and oblique muscles, although less is known about the structure of invertebrate connectins at present. The protein that maintains such a structure is present not only in vertebrate striated muscle, but also in invertebrate striated and oblique muscle. In the present study, we analyzed the partial primary structure of a 1200K protein, which is a connectin-like protein that is expressed in Neanthes sp. body wall muscle that is in turn composed of oblique muscle. Antibody screening of a cDNA library of Neanthes sp. body wall muscle identified two different clones. Both clones coded for a sequence predominantly comprised of the four amino acids proline (P), glutamate (E), valine (V) and lysine (K). One clone included a PEVK like repeat sequence flanked by an Ig domain, while the other clone comprised a distinct 14 amino acid repeat rich in PEVK residues, flanked by a non-repetitive unique sequence. The PEVK region is found in vertebrate connectin and is thought to generate elasticity and be responsible for passive tension of the muscle. The antibodies produced against a portion of each clone both reacted with bands corresponding to 1200 kDa present in Neanthes sp. body wall muscle. Therefore, our results demonstrate that this 1200K-protein is a connectin-like elastic protein and includes specific PEVK-like fragment. We suggest that this 1200K protein plays a major role in maintaining the structure of oblique muscle in invertebrates. PMID- 16470336 TI - Muscle atrophy in titin M-line deficient mice. AB - We investigated the response to deletion of the titin M-line region in striated muscle, using a titin knockout model and a range of techniques that include histology, in situ hybridization, electron microscopy, and 2D gel analysis. We found that the loss of titin's kinase domain and binding sites for myomesin and MURF-1 causes structural changes in the sarcomere that proceed from the M-line to the Z-disc and ultimately result in disassembly of the sarcomere. Disassembly goes along with central localization of nuclei (a hallmark for muscular dystrophy), up-regulation of heat-shock proteins, and induction of proteasome activity. While fiber type composition does not change in soleus and extensor digitorum longus muscle, fiber size is reduced. Animals die from complications of muscle atrophy at five weeks of age. In addition to the structural importance of the titin M-line region in any striated muscle, our data show how differences in M-line composition between heart and skeletal muscle affect sarcomere stability and function. PMID- 16470337 TI - The sarcomeric M-band during development and in disease. AB - The C-terminus of connectin/titin at the M-band of the sarcomere interacts with several structural as well as potential signalling proteins. One of these is myomesin, which can also bind to myosin and has been suggested to function as an integral structural linker of the thick filaments into the sarcomere. Recent evidence that myomesin possesses the ability to form antiparallel dimers via its C-terminal domain has prompted us to propose a novel three-dimensional model for the sarcomeric M-band. A splice variant of myomesin, termed EH-myomesin, contains an additional segment that has disordered conformation and functions as an entropic spring. It is expressed in a subset of muscle types that are characterised by a broader operational range and are more resistant to damage caused by eccentric contraction. In addition, it is also re-expressed in dilated cardiomyopathy. DRAL/FHL-2 is another protein that interacts with the M-band portion of connectin/titin and which probably functions as an adaptor for the compartmentalisation of metabolic enzymes. Together these results suggest that the M-band is crucial for sarcomere function and maintenance and that its molecular composition can be adapted to divergent physiological needs in different muscle types, which may help to cope with pathological alterations. PMID- 16470338 TI - The chromatin regulatory code: beyond a histone code. AB - In this commentary on the contribution by Arndt Benecke in this issue, I discuss why the notion of "chromatin code" introduced and elaborated in this paper is to be preferred to that of "histone code". Speaking of a code as regards nucleosome conformation and histone tail post-translational modifications only makes sense within the chromatin fiber, where their physico-chemical features can be translated into regulatory programs at the genome level, by means of a complex, multi-level interplay with the fiber architecture and dynamics settled in the course of Evolution. In particular, this chromatin code presumably exploits allosteric transitions of the chromatin fiber. The chromatin structure dependence of its translation suggests two alternative modes of transcription initiation regulation, also proposed in the paper by A. Benecke in this issue for interpreting strikingly bimodal micro-array data. PMID- 16470339 TI - Regulating chromatin: on code and dynamic models. AB - In this short comment on the contributions by Arndt Benecke, Francois Fuks and Helmut Schiessel I refer to some recent experimental work which sheds light on the histone code hypothesis. Further, I make some remarks on the role theoretical approaches can play in decoding chromatin regulation. PMID- 16470341 TI - Variants of Coconut cadang-cadang viroid isolated from an African oil palm (Elaies guineensis Jacq.) in Malaysia. AB - Variants of Coconut cadang-cadang viroid have been identified in a plantation oil palm growing in Malaysia. Three size classes are described, comprising 297, 293, and 270 nt. Compared with the 296-nt form of coconut cadang-cadang viroid (CCCVd), all variants substituted C31 --> U in the pathogenicity domain and A175 -> C in the right-hand terminus. Other mutations and deletions accounted for the different sizes. These are the first sequences reported for variants of Coconut cadang-cadang viroid in a species other than coconut palm, and this is the first evidence that variants closely related to CCCVd occur outside the Philippines. PMID- 16470340 TI - Antipsychotics in early onset Schizophrenia: Systematic review and meta-analysis. AB - BACKGROUND: To develop an evidence base for using antipsychotic medications for schizophrenia in children and adolescents. METHOD: Data sources were identified in PsychINFO (1872-2003), MEDLINE (1966-2003), and articles in reference lists. Study selection criteria: (1) treatment with antipsychotics; (2) ages were between 5 and 18 years; (3) sample diagnosed with schizophrenia; (4) prospective design; (5) rating instruments used. Fifteen studies met inclusion criteria and were rated. Study quality was independently rated. RESULTS: Average response rate among 8 studies employing atypicals was 55.7% compared to 72.3% among 13 studies employing typicals. The difference was statistically different at the trend level (z = 1.65, P < 0.10). The effect size on a continuous measure was 0.36 in favor of typicals. When study quality was included in the model, the effect of medication type remained unchanged. Average weight gain in patients treated with typicals was 1.4 Kg. compared to 4.5 Kg for those treated with atypicals. Sedation was more common among those on atypicals. The rate of extrapyramidal side effects was similar among the two groups CONCLUSIONS: Antipsychotic medications seem effective for schizophrenia treatment in children and adolescents. Typicals appear to be more effective and cause less weight gain than atypicals. However, more rigorous clinical trials are necessary. PMID- 16470342 TI - 3D reconstruction of the diaphragm for virtual traumatology. AB - This study lies within the scope of passive road safety, and more particularly injury mechanisms of the abdominal area. The finite element modeling, which makes it possible to simulate a road accident and to observe the possible bone fractures or internal tissue injuries, allows large projections in the comprehension of injury mechanisms. However, the digital models already available and used in accidentology do not offer as one very simplified description of the diaphragm, as well for its geometry as for its bracing aspect and the modifications that this could induce in the behavior of abdominal organs and vessels at impact. In order to develop an accurate model of diaphragm for road safety research, a 3D reconstruction was performed, based on a sitting post mortem Human subject sections. The resulting geometry was then turned into a segmented mechanical component (using the finite element method) and included in a full human model already available. The result is a valuable tool to improve the knowledge of injury mechanisms involved in car crashes at the abdominal level. PMID- 16470343 TI - Morphological peculiarities of the deep infratemporal fossa in advanced age. AB - The main osseous landmarks of the border area between the infratemporal fossa and the para- and retro pharyngeal space are the sphenoidal spine and the pterygoid and styloid processes, and the styloid vagina. These landmarks, as well as some vascular anomalies, were studied in order to illustrate the variable anatomy, which is encountered in the surgical lateral transzygomatic infratemporal fossa approach. Hundred well-preserved human skull bases were examined. The deep infratemporal fossa was dissected into 54 halves of fixed cadaveric head and neck specimens of both sexes. Dry skull specimens of New and Old World monkeys, skulls of rodents, herbivora and carnivora, and computed tomograms of the head of Macaca fuscata Japonica, were also studied. In 91 of the human skull bases, the sphenoidal spine was prominent and well developed. In three skulls, the spine was absent. In four specimens, however, the spine was not sphenoidal but a part of the temporal bone, occurring in the form of a process, which emanated from the styloid vagina. In two further cases, there was unilaterally a duplicated spine; the anterior part represented a regular sphenoidal spine, while the posterior part constituted a part of the vagina of the styloid process. A complete osseous bar, arch or lamina-connecting the posterior border of the lateral lamina of the pterygoid process and the sphenoidal spine-existed in six of the human dry skulls. In ten of the human skulls examined, the breadth of the lateral lamina of the pterygoid process was greater than 10 mm; thus, the so-called pterygospinous (ps) and the pterygostyloid gates-of significance where surgical approaches are concerned-were less than 10 mm in width. Fibrous or muscular connections were also found in some cadaveric specimens between the posterior border of the lateral lamina of the pterygoid process and the sphenoidal spine: a ps ligament existed in 11 cases (20.4%) and a ps muscle in 5 cases (9.2 %), in 3 of which it inserted into both the medial wall capsule and the articular disc of the temporo mandibular joint. Among the cadaveric specimens exhibiting ps structures was one in which an osseous ps bar occurred together with a ps muscle; in two cases a strong ps ligament was observed together with a ps muscle. The distribution pattern of the mandibular nerve was affected by the positioning of the ps bar, ligament and muscle when the latter were present. The existence of a wide ps bar was noted in all the skulls of herbivora, rodentia, carnivora, and Old World monkeys that were examined, but never in those of the New World monkeys; it is likely that, in the human, this ps bar represents a phylogenetic remnant. In the human dry skull specimens and cadaveric material, the ps ligament was found to be a reinforcement of the interpterygoid fascia, and the ps muscle to be a third head of the lateral pterygoid muscle. In two cases, the internal carotid artery exhibited a significant elongation and space-consuming tortuosity (so-called coiling behavior) in the depth of the infratemporal fossa. PMID- 16470344 TI - A model of disease-specific worry in heritable disease: the influence of family history, perceived risk and worry about other illnesses. AB - Disease-related worry is associated with family history and perceived risk of that disease; however, the influences of general risk perceptions and tendencies to worry about diseases have been neglected in the literature. This study investigates a model of disease-specific worry which includes family history, disease-specific perceived risk, and perceived risk for and worry about other diseases. Participants completed a survey assessing these variables in relation to several heritable diseases. Structural equation modeling found that family history predicted disease-specific perceived risk but not perceived risk for other diseases. Disease-specific perceived risk predicted disease-specific worry and worry about other diseases. Perceived risk for other diseases predicted worry about other diseases and disease-specific perceived risk but not disease-specific worry. Disease-specific worry predicted worry about other diseases. This model was supported across several diseases and indicates that disease-specific and general considerations of risk influence worry about a disease and should be considered in interventions. PMID- 16470346 TI - Development of congenic rat strains for alcohol consumption derived from the alcohol-preferring and nonpreferring rats. AB - A genome scan of the F2 generation from an inbred alcohol-preferring (iP) and inbred alcohol-nonpreferring (iNP) rat cross identified a significant quantitative trait locus (QTL) on chromosome 4 with a lod score of 9.2. To confirm this QTL and to create animals for fine mapping of the QTL region, chromosome 4 reciprocal congenic strains were developed by transferring the chromosome 4 QTL interval into the respective iP or iNP backgrounds. The iP strain was crossed with the iNP strain to create iPiNP F1 animals, which were backcrossed to either iNP or iP animals to produce the N2 generation. Using marker-assisted selection, 10 generations of backcrossing were performed. The selection was followed by an intercross between the N10 animals to produce homozygous animals (N10F1), resulting in the finished congenic strains. Congenic strains in which the iP chromosome 4 QTL interval was transferred to the iNP (NP.P) and the iNP chromosome 4 QTL was transferred to the iP (P.NP) exhibited the expected effect on alcohol consumption of the donor strain. Development of these congenic strains further indicates that the chromosome 4 QTL region is, in part, responsible for the disparate alcohol consumption observed between the iP and iNP rats. These congenic animals will be an invaluable resource for fine mapping the QTL region and for the identification of the gene(s) that influences the drinking behavior of the iP and iNP rats. PMID- 16470345 TI - Conceptualizing and measuring ethnic discrimination in health research. AB - This paper presents the General Ethnic Discrimination Scale, an 18-item measure of perceived ethnic discrimination that can be used in health research with any ethnic group. The 1569 participants (half college students, half community adults) completed the General Ethnic Discrimination scale and measures of cigarette smoking and of psychiatric symptoms. Results revealed that the General Ethnic Discrimination subscales model the latent construct of perceived ethnic discrimination equally well for Blacks, Latinos, Asians, and Whites. Discrimination was strongly related to psychiatric symptoms and to current cigarette smoking for ethnic minorities and Whites alike, but such relationships were stronger for ethnic minorities. Minorities who experienced frequent discrimination were 2.3 times more likely than their low-discrimination counterparts to be smokers. This 5th grade reading-level scale takes 10 min to complete and has sufficient, initial psychometric integrity for use in clinical and community health studies. PMID- 16470347 TI - A molecular thermometer based on the delayed fluorescence of C70 dispersed in a polystyrene film. AB - A new optical molecular thermometer, based on the thermally activated delayed fluorescence of C70 dispersed in a polystyrene film, was developed. In the presence of oxygen, the fluorescence intensity of the C70 film is essentially temperature independent in a wide range. In the absence of oxygen, however, the fluorescence intensity markedly increases with temperature. At room temperature (25 degrees C), and after degassing the sample, the fluorescence intensity of C70 increases 22 times, while at 100 degrees C the fluorescence intensity is increased by 70 times. With our system, the very weak fluorescence of C70 (phi(F) congruent with 5 x 10(-4), in toluene) can be increased up to 91 times (up to an estimated maximum value phi(F) = 0.046). The estimate value of the singlet triplet gap (29 kJ mol(-1)) and the fluorescence lifetime (0.63 ns) of the C70 in film are in agreement with the values reported in the literature for C70 in solution. The values of the phosphorescence lifetime at room temperature (23 ms) and the quantum yield of triplet formation (0.989) were also determined. The system is completely reversible with respect to heating-cooling cycles. PMID- 16470349 TI - Comparison of trifluoroacetyl monostyryl and distyryl dyes: effects of chromophore elongation on the spectral properties and chemical reactivity. AB - The trifluoroacetyl distyryl derivative 1-[4-(2-[4-[2-(4-dibutylaminophenyl) vinyl]-phenyl]-vinyl)-phenyl]-2,2,2-trifluoroethanone was compared with the related monostyryl derivative 1-[4-[2-(4-dibutylaminophenyl)-vinyl]-phenyl]-2,2,2 trifluoroethanone with respect to spectral properties and sensitivity to amines. Both trifluoroacetyl derivatives had their absorbance maximum at around 445 nm. The fluorescence of the distyryl dye, however, was observed at significantly longer wavelengths than that of the monostyryl dye, indicating the effect of structural extension of the chromophore system. Furthermore, the distyryl dye exhibited significantly smaller quantum yields in polar solvents than the monostyryl dye. Both dyes were capable of chemically reacting with amines in that their trifluoroacetyl function was converted into a hemiaminal. Consequently, absorbance and fluorescence of both dyes were shifted to shorter wavelengths. The positions of the fluorescence maxima of the spectra when converting from trifluoroacetyl to hemiaminal form were shifted by an almost identical amount for both mono- and distyryl derivative. The hemiaminal form of the distyryl derivative, however, exhibited much larger quantum yields in both polar and nonpolar solvents than the hemiaminal form of the monostyryl dye. The structural extension of the chromophore affected the sensitivity to amines by enhancing the chemical reactivity of the distyryl dye over the monostyryl derivative. PMID- 16470348 TI - Fluorescence study of dehydroabietic acid-based bipolar arylamine-quinoxalines. AB - The absorption and fluorescence spectra, lifetimes and quantum yields of a series of triarylaminequinoxaline bipolar compounds, with and without the bulky dehydroabietic acid group, have been studied in toluene solution. This bulky group is introduced to improve solubility and thermal properties of these systems. It is shown that this does not affect their spectral or photophysical behavior. The compounds show relatively strong fluorescence, with the emission maximum strongly dependent upon the substituents present. Oxidation potentials have also been determined in acetonitrile solution, and again indicate that introduction of the resin acid moiety has no effect on these properties. PMID- 16470350 TI - Application of fluorescence with polarized light to evaluate the orientation of dyes adsorbed in layered materials. AB - A new method of fluorescence polarization is applied to evaluate the angle of the preferential orientation of Rhodamine 6G (R6G) dye adsorbed in supported thin films of Laponite (Lap) clay. The method is based in the determination of the fluorescence dichroic ratio, obtained from the recorded fluorescence spectra with the detection polarizer horizontally and vertically oriented, as a function of the twisted angle of the film around its vertical axis, keeping the excitation polarizer in a fixed direction. The validity of the method is checked by comparing the experimental results obtained with both vertically and horizontally polarized excitations to that previously provided by absorption spectroscopy with linearly polarized light. A preferential orientation angle with respect to the normal to the clay layer of 62 degrees is derived for R6G monomers adsorbed in Lap films. PMID- 16470351 TI - Electronic excitation energy migration in partly ordered polymeric films. AB - Multistep intermolecular energy migration between elongated fluorophores (carbocyanines) in uniaxially oriented polymer films is studied based on fluorescence depolarization and Monte-Carlo simulations. Contrary to disordered systems it is found experimentally that the concentration depolarization of fluorescence in uniaxially oriented films is extremely weak despite effective energy migration. Based on the concentration depolarization experiment in the ordered matrix it is possible to estimate the angle between absorption and fluorescence transition moments of carbocyanines. The values of that angle are very close to those obtained from other methods. PMID- 16470352 TI - High risk of hypogonadism after traumatic brain injury: clinical implications. AB - Several recent studies have convincingly documented a close association between traumatic brain injury (TBI) and pituitary dysfunction. Post-traumatic hypogonadism is very common in the acute post-TBI phase, though most cases recover within six to twelve months following trauma. The functional significance of early hypogonadism, which may reflect adaptation to acute illness, is not known. Hypogonadism persists, however, in 10-17% of long-term survivors. Sex steroid deficiency has implications beyond psychosexual function and fertility for survivors of TBI. Muscle weakness may impair functional recovery from trauma and osteoporosis may be exacerbated by immobility secondary to trauma. Identification and appropriate and timely management of post-traumatic hypogonadism is important in order to optimise patient recovery from head trauma, improve quality of life and avoid the long-term adverse consequences of untreated sex steroid deficiency. PMID- 16470353 TI - Purification and characterization of a fibrinolytic subtilisin-like protease of Bacillus subtilis TP-6 from an Indonesian fermented soybean, Tempeh. AB - We have isolated a bacterium (TP-6) from the Indonesian fermented soybean, Tempeh, which produces a strong fibrinolytic protease and was identified as Bacillus subtilis. The protease (TPase) was purified to homogeneity by ammonium sulfate fractionation and octyl sepharose and SP sepharose chromatography. The N terminal amino acid sequence of the 27.5 kDa enzyme was determined, and the encoding gene was cloned and sequenced. The result demonstrates that TPase is a serine protease of the subtilisin family consisting of 275 amino acid residues in its mature form. Its apparent K (m) and V (max) for the synthetic substrate N succinyl-Ala-Ala-Pro-Phe-pNA were 259 microM and 145 micromol mg(-1) min(-1), respectively. The fibrinogen degradation pattern generated by TPase as a function of time was similar to that obtained with plasmin. In addition, N-terminal amino acid sequence analysis of the fibrinogen degradation products demonstrated that TPase cleaves Glu (or Asp) near hydrophobic acids as a P1 site in the alpha- and beta-chains of fibrinogen to generate fragments D', E', and D' similar to those generated by plasmin. On plasminogen-rich fibrin plates, TPase did not seem to activate fibrin clot lysis. Moreover, the enzyme converted the active plasminogen activator inhibitor-1 to the latent form. PMID- 16470354 TI - [Sequelae of organ transplantation]. AB - Transplant medicine has significantly changed the prognosis of diseases leading to terminal organ failure. It has evolved from an experimental procedure to standard therapy for liver, kidney and cardio-vascular diseases. Transplant medicine combines operative organ replacement with the management of severely ill patients before transplantation, as well as life-long follow-up of organ graft recipients. Ten year survival rates of 65% to over 90% have led to a steady increase of transplanted patients seen by general medical care providers which represents a challenge for practicing internists. Apart from organ-specific conditions, infectious, immunosuppressant-associated and metabolic consequences determine long-term survival. These include virus reactivation, graft rejection, anastomotic problems but more importantly general mortality determining factors such as diabetes, renal insufficiency and hypertension, which are often a consequence of immunosuppressant administration. They directly impact long-term survival. The awareness and treatment of these secondary conditions of organ transplantation in routine medical practice contributes significantly to secure the long term success of transplant medicine. PMID- 16470355 TI - [Late effects after chemotherapy]. AB - Today, about 60% of adult patients and 80% children diagnosed with cancer will survive 5 years after diagnosis. Approximately two thirds of cancer survivors will experience at least one late effect, and about one third severe or life threatening late effects. The aim of cancer treatment today is to cure the malignant disease but at the same time, to minimize the incidence of post treatment complications. In the current overview we summarize, based on the most recent publications, typical late effects in cancer survivors. PMID- 16470356 TI - [Renal anemia - an important secondary disease in renal insufficiency]. AB - Anemia is as a frequent complication in patients with chronic kidney disease, which gains in importance in the treatment of patients with renal disease. The main cause of renal anemia is the inadequately low production of endogenous erythropoietin. Often the patients develop an additional absolute or functional iron deficiency, which complicates the diagnostic and therapeutic procedures. Substitution of recombinant human erythropoietin (r-HuEPO) is the most effective therapy. The goal is a stable haemoglobin level >11 g/dl. An often additional existing iron deficiency should be balanced adequately according to the guidelines. With consequent and early treatment morbidity, mortality, and quality of life can be effectively improved. PMID- 16470357 TI - [Sequelae of hypertenson: kidney disease]. AB - Nephropathy is one of the frequent sequelae of hypertension. Arterial hypertension causes both arterial and capillary changes in the kidneys as well as development of interstitial fibrosis. Systemic or intraglomerular pressure increase leads to albuminuria and proteinuria, which in turn contributes to further damage of the kidneys. Impairment of the kidneys due to hypertension can ultimately result in terminal renal insufficiency necessitating dialysis. Metabolic factors such as hyperlipidemia, hyperuricemia, hyperhomocysteinemia, and insulin resistance can aggravate renal lesions. Effective lowering of blood pressure in conjunction with management of the metabolic risk factors is decisive for prevention of chronic kidney disease, which in turn must be considered a factor involved in exacerbation of the hypertension. PMID- 16470359 TI - [62 year-old patient with rapid progressive edema, low potassium and hypertension]. AB - A 62 year-old patient was admitted to hospital with rapid progressive edema, low potassium and hypertension. This symptoms are caused by Cushing's syndrome through ectopic paraneoplastic ACTH-production. Primary neoplasm is a small cell cancer. A Sertoli-cell-tumor of the testis was diagnosed as an additional carcinoma. Palliative chemotherapy and adrenostatic agents did not improve the clinical findings and the patient died eight weeks after admission. PMID- 16470360 TI - [Therapy of hypercalcemia with ibandronate in case of acute renal failure]. AB - Hypercalcaemia is a common complication of malignancies associated with bone destruction. Besides, benign diseases as sarcoidosis or hyperparathyroidism may lead to hypercalcaemia. The main principles of modern therapy contain a forced diuresis as well as the application of bisphosphonates. Latter substances bear the danger of developing a renal insufficiency. Here, we report the case of a female patient, suffering from primary hyperparathyroidism with severe hypercalcaemia and calcium levels up to 6 mmol/l, who developed acute renal failure. We treated the patient with forced diuresis and repeated infusions of ibandronate (5 x 6 mg ibandronate). Even if lowering the serum levels of calcium only for a short time after each application, yet we could improve renal function by these means. Only after performing a parathyroidectomy, we could see a sustained decline of calcium levels. This case report supports the results of other publications, that have reported the missing nephrotoxic effect of ibandronate compared to other bisphosphonates. PMID- 16470361 TI - Dexamethasone treatment in adults with pneumococcal meningitis: risk factors for death. AB - In experimental meningitis, adjunctive treatment with steroids reduces cerebrospinal fluid inflammation and thereby improves neurological outcome. On the basis of these findings, several clinical trials have assessed treatment with adjunctive steroids in bacterial meningitis, with conflicting results. Recently, the results of the European Dexamethasone Study showed a beneficial effect of adjunctive dexamethasone in adults with bacterial meningitis. In that study, the effect of dexamethasone on outcome was most striking in patients with pneumococcal meningitis. The aim of the present study was to further evaluate the effect of dexamethasone in adults with pneumococcal meningitis by performing a post hoc analysis of the European Dexamethasone Study. In a multivariate analysis, tachycardia (p=0.02), advanced age (p=0.03), low score on the Glasgow Coma Scale (p=0.03), positive blood culture (p=0.04), and absence of dexamethasone therapy (p=0.05) were independent predictors for death. Patients who were treated with adjunctive dexamethasone were less likely to develop both systemic and neurological complications during hospitalisation, compared with patients who received placebo. In conclusion, independent risk factors for death in pneumococcal meningitis are tachycardia, advanced age, low level of consciousness, bacteraemia, and absence of dexamethasone therapy. Treatment with adjunctive dexamethasone in adults with pneumococcal meningitis reduces both systemic and neurological complications. PMID- 16470362 TI - Gene expression profiling of bovine in vitro adipogenesis using a cDNA microarray. AB - The gene expression profile of bovine bone marrow stromal cells undergoing adipogenesis was established using a custom cDNA microarray. Cells that were treated with adipogenic stimulants and those that were not were collected at each of the six time points, and gene expression differences between the treated and untreated samples within each time point were compared using a microarray. Statistical analyses revealed that 158 genes showed a minimum fold change of 2 in at least one of the five post-differentiation time points. These genes are involved in various cellular pathways and functions, including lipogenesis, glycolysis, cytoskeleton remodelling, extracellular matrix, transcription as well as various signalling pathways such as insulin, calcium and wingless signalling. The experiment also identified 17 differentially expressed (DE) microarray elements with no assigned function. Quantitative real-time PCR was employed to validate eight DE genes, and the PCR data were found to reproduce the microarray data for these eight genes. Subsequent gene ontology annotation was able to provide a global overview of the molecular function of DE genes during adipogenesis. This analysis was able to indicate the importance of different gene categories at various stages of adipogenic conversion, thereby providing further insights into the molecular changes during bovine adipogenesis. PMID- 16470363 TI - Applications of real-time polymerase chain reaction for quantification of microorganisms in environmental samples. AB - Due to the advanced development of fluorogenic chemistry, quantitative real-time polymerase chain reaction (qRT-PCR) has become an emerging technique for the detection and quantification of microorganisms in the environment. Compared with the conventional hybridization- and PCR-based techniques, qRT-PCR not only has better sensitivity and reproducibility, but it is also quicker to perform and has a minimum risk of amplicon carryover contamination. This article reviews the principle of this emerging technique, its detection reagents, target DNAs, quantification procedures, and affecting factors. The applications of qRT-PCR for the quantification of microorganisms in the environment are also summarized. PMID- 16470364 TI - Cloning of beta-1,3-1,4-glucanase gene from Bacillus licheniformis EGW039 (CGMCC 0635) and its expression in Escherichia coli BL21 (DE3). AB - Beta-1,3-1,4-glucanase has been applied in the brewing and animal feed additive industry. It can effectively improve digestibility of barley-based diets and reduce enteritis. It also reduces viscosity during mashing for high-quality brewers malt. The aim of this work is to clone beta-1,3-1,4-glucanase-encoding gene and express it heterogeneously. The gene was amplified by polymerase chain reaction using Bacillus licheniformis genomic DNA as the template and ligated into the expression vector pET28a. The recombinant vector was transformed into Escherichia coli. The estimated molecular weight of the recombinant enzyme with a six-His tag at the N terminus was about 28 kDa, and its activities in cell lysate supernatant were 1,286 and 986 U ml(-1) for 1% (w/v) barley beta-glucan and 1% (w/v) lichenan, respectively. Accordingly, the specific activities were 2,479 and 1,906 U mg(-1) for these two substrates. The expression level of recombinant beta 1,3-1,4-glucanase was about 60.9% of the total protein and about 12.5% of the total soluble protein in crude cell lysate supernatant. Acidity and temperature optimal for this recombinant enzyme was pH 5.6 and 40 degrees C, respectively. PMID- 16470365 TI - Characterization of Melanocarpus albomyces steryl esterase produced in Trichoderma reesei and modification of fibre products with the enzyme. AB - Melanocarpus albomyces steryl esterase STE1 is considered to be an interesting tool for several industrial applications due to its broad substrate specificity. STE1 was produced in the filamentous fungus Trichoderma reesei in a laboratory bioreactor at an estimated production level of 280 mg l(-l). The properties of the purified recombinant enzyme (rSTE1), such as substrate specificity, molecular mass, pH optimum and stability and thermostability, were characterized and compared to the corresponding properties of the native enzyme. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis showed one band with a molecular weight of 60 kDa for rSTE1, whereas analytical gel filtration showed a dimeric structure with a molecular weight of 120 kDa. The rSTE1 was somewhat less stable under different conditions and had slightly lower activities on various substrates than the native STE1. The effects of rSTE1 on the properties of paper sheets and polyethylene terephthalate (PET) fabric were preliminarily evaluated. Due to the hydrolysis of triglycerides and steryl esters by the rSTE1 treatment, the tensile strength and hydrophilicity of the paper were increased. The rSTE1 treatment increased significantly the polarity of PET by hydrolysing the ester bonds in the polyester backbone. Dyeing of PET with methylene blue was also slightly improved after rSTE1 treatment. PMID- 16470367 TI - Effect of spatial distribution of magnetic dipoles on Lamor frequency distribution and MR Signal decay--a numerical approach under static dephasing conditions. AB - Cells loaded with superparamagnetic iron oxide (SPIO) cause relatively strong magnetic field distortions, implying that field position effects of neighboring SPIO loaded cells have to be accounted for. We treated SPIO loaded cells as magnetic dipoles in a homogeneous magnetic field and computed the 3D frequency distribution and the related signal decay using a numerical approach under static dephasing conditions. The volume fraction of dipoles was kept constant for all simulations. For larger randomly distributed magnetic dipoles we found a non Lorentzian frequency distribution and a non-monoexponential signal decay whereas, for smaller dipoles, the frequency distribution was more Lorentzian and the signal decay was well fitted monoexponentially. Moreover, based on our numerical and experimental findings, we found the gradient echo signal decay due to a single SPIO labeled cell to be non-monoexponential. The numerical approach provides deeper understanding of how the spatial distribution of SPIO loaded cells affects the MR signal decay. This fact has to be considered for the in vivo quantification of SPIO loaded cells, implying that in tissues with different spatial distributions of identical SPIO concentrations, different signal decays might be observed. PMID- 16470366 TI - Sequencing of the intergenic 16S-23S rRNA spacer (ITS) region of Mollicutes species and their identification using microarray-based assay and DNA sequencing. AB - We have completed sequencing the 16S-23S rRNA intergenic transcribed spacer (ITS) region of most known Mycoplasma , Acholeplasma , Ureaplasma , Mesoplasma , and Spiroplasma species. Analysis of the sequence data revealed a significant interspecies variability and low intraspecies polymorphism of the ITS region among Mollicutes . This finding enabled the application of a combined polymerase chain reaction-microarray technology for identifying Mollicutes species. The microarray included individual species-specific oligonucleotide probes for characterizing human Mollicutes species and other species known to be common cell line contaminants. Evaluation of the microarray was conducted using multiple, previously characterized, Mollicutes species. The microarray analysis of the samples used demonstrated a highly specific assay, which is capable of rapid and accurate discrimination among Mollicutes species. PMID- 16470368 TI - Scintigraphic appearance of "linitis plastica" in a patient with gastric non Hodgkin's lymphoma on 18F-FDG imaging. PMID- 16470369 TI - Significant clinical impact and prognostic stratification provided by FDG-PET in the staging of oesophageal cancer. AB - PURPOSE: To evaluate the clinical impact of FDG-PET in staging oesophageal cancer and whether this information improves prognostic stratification. METHODS: Impact was based on comparison of a prospectively recorded pre-PET plan with post-PET treatment in 68 consecutive patients undergoing primary staging. Survival was analysed using the Kaplan-Meier product limit method and the Cox proportional hazards regression model. RESULTS: FDG-PET findings impacted on the management of 27/68 patients (40%): in 12 therapy was changed from curative to palliative and in three from palliative to curative, while in 12 other patients there was a change in the treatment modality or delivery but not in the treatment intent. The median survival was 21 months, with post-PET stage and treatment intent both strongly associated with survival (p<0.001). Conventional stage was not able to clearly stratify this population. CONCLUSION: The use of FDG-PET for primary staging of oesophageal cancer changed the clinical management of more than one third of patients and provided superior prognostic stratification compared with conventional investigations. PMID- 16470370 TI - Fully automatic diagnostic system for early- and late-onset mild Alzheimer's disease using FDG PET and 3D-SSP. AB - PURPOSE: The purpose of this study was to design a fully automatic computer assisted diagnostic system for early- and late-onset mild Alzheimer's disease (AD). METHODS: Glucose metabolic images were obtained from mild AD patients and normal controls using positron emission tomography (PET) and( 18)F fluorodeoxyglucose (FDG). Two groups of 20 mild AD patients with different ages of onset were examined. A fully automatic diagnostic system using the statistical brain mapping method was established from the early-onset (EO) and late-onset (LO) groups, with mean ages of 59.1 and 70.9 years and mean MMSE scores of 23.3 and 22.8, respectively. Aged-matched normal subjects were used as controls. We compared the diagnostic performance of visual inspection of conventional axial FDG PET images by experts and beginners with that of our fully automatic diagnostic system in another 15 EO and 15 LO AD patients (mean age 58.4 and 71.7, mean MMSE 23.6 and 23.1, respectively) and 30 age-matched normal controls. A receiver operating characteristic (ROC) analysis was performed to compare data. RESULTS: The diagnostic performance of the automatic diagnostic system was comparable with that of visual inspection by experts. The area under the ROC curve for the automatic diagnostic system was 0.967 for EO AD patients and 0.878 for LO AD patients. The mean area under the ROC curve for visual inspection by experts was 0.863 and 0.881 for the EO and LO AD patients, respectively. The mean area under the ROC curve for visual inspection by beginners was 0.828 and 0.717, respectively. CONCLUSION: The fully automatic diagnostic system for EO and LO AD was able to perform at a similar diagnostic level to visual inspection of conventional axial images by experts. PMID- 16470371 TI - Mutagenesis of conserved charged amino acids in SLH domains of Thermoanaerobacterium thermosulfurigenes EM1 affects attachment to cell wall sacculi. AB - SLH domains (for surface layer homology) are involved in the attachment of proteins to bacterial cell walls. The data presented here assign the conserved TRAE motif within SLH domains a key role for the binding. The charged amino acids arginine (R) or/and glutamic acid (E) were replaced via site-directed mutagenesis by different amino acids. Effects were visualized in an in vitro binding assay using native cell wall sacculi of Thermoanaerobacterium thermosulfurigenes EM1 and different variants of an SLH protein which consisted of the triplicate SLH domain of xylanase XynA of this bacterium and which was purified after expression in Escherichia coli. The results indicated (1) that the TRAE motif is critical for the binding function of SLH domains, (2) that a functional TRAE motif is necessary in all three domains, (3) that a least one (preferentially positively) charged amino acid in the TRAE motif is required for the functionality of the SLH domain, and (4) that the position of the negatively and positively charged amino acids is important. The finding that the cell wall of T. thermosulfurigenes EM1 contains pyruvate (4 microg mg(-1)) is in agreement with the hypothesis that pyruvylated secondary cell wall polymers function as ligand for SLH domains. PMID- 16470372 TI - Anaerobic cells of Bacillus cereus F4430/73 respond to low oxidoreduction potential by metabolic readjustments and activation of enterotoxin expression. AB - In the present study, a food-borne pathogen strain of Bacillus cereus (F4430/73) was anaerobically grown in controlled-batch conditions under low initial oxidoreduction potential (ORP=-148 mV) using hydrogen gas as reducing agent. Its physiological characteristics, including growth, glucose fermentation capacity and enterotoxin production, were compared with anaerobic conditions generated by nitrogen gas (ORP=+ 45 mV). The results showed that low ORP affected growth mainly during the early stages. Maximal specific rates of growth and glucose consumption were reduced, and drastic changes in time profiles of fermentation product concentration were observed. Production of lactate was promoted at the expense of acetate. Nevertheless, low ORP did not affect final biomass yield. Under both ORP conditions, Non-haemolytic enterotoxin (Nhe) was produced early during the exponential growth phase as a first enterotoxin and Haemolysin BL (Hbl) later during the early stationary growth phase as a second enterotoxin. The major effect of low ORP was the strong stimulation of Hbl production and, to a lesser extent, Nhe production. This control was complex, involving different levels of regulation. We discussed the regulation of enterotoxin expression and the involvement of the pleiotropic regulator PlcR. PMID- 16470373 TI - [Lucio phenomenon. Vasculitis or occlusive vasculopathy?]. AB - Lucio's phenomenon (LP) occurs in patients with Lucio leprosy (LuL). Some interpret the cutaneous lesions and their histopathology as a thrombotic/occlusive condition, while others consider it leukocytoclastic vasculitis. The clinical similarities between the cutaneous manifestations of LP and antiphospholipid syndrome (APS) led us to investigate the relationship between these two pathological conditions. We studied the clinical, laboratory and histopathologic aspects of LuL and LP in one patient and compare these results to APS. The examination of antiphospholipid antibodies showed positive anticardiolipin (aCL) and lupus anticoagulant (LAC). The histopathological slides of cutaneous biopsies were stained by hematoxylin-eosin and Fite-Faraco. They showed the typical features of LuL, as well as thrombi, endothelial proliferation, vessel wall thickening and obliteration of the lumen. Leukocytoclastic vasculitis was not found. The clinical pattern of LuL in our case is identical to that described by Lucio and Latapi. The necrotic lesions of LP in our patient resembled APS. This suggests that LP could be considered APS secondary to LuL. Multidrug treatment for multibacillary patients (MDT-MP) was successful, with no need for thalidomide or systemic corticosteroids. PMID- 16470374 TI - The desirability and feasibility of economic studies of drugs post-launch. PMID- 16470375 TI - Clinical relevance of diffusion and perfusion magnetic resonance imaging in assessing intra-axial brain tumors. AB - Advanced magnetic resonance (MR) imaging techniques provide physiologic information that complements the anatomic information available from conventional MR imaging. We evaluated the roles of diffusion and perfusion imaging for the assessment of grade and type of histologically proven intraaxial brain tumors. A total of 28 patients with intraaxial brain tumors underwent conventional MR imaging (T2- and T1-weighted sequences after gadobenate dimeglumine injection), diffusion imaging and T2*-weighted echo-planar perfusion imaging. Examinations were performed on 19 patients during initial diagnosis and on nine patients during follow-up therapy. Determinations of relative cerebral blood volume (rCBV) and apparent diffusion coefficient (ADC) were performed in the solid parts of each tumor, peritumoral region and contralateral white matter. For gliomas, rCBV values were greater in high-grade than in low-grade tumors (3.87+/-1.94 versus 1.30+/-0.42) at the time of initial diagnosis. rCBV values were increased in all recurrent tumors, except in one patient who presented with a combination of recurrent glioblastoma and massive radionecrosis on histology. Low-grade gliomas had low rCBV even in the presence of contrast medium enhancement. Differentiation between high- and low-grade gliomas was not possible using diffusion-weighted images and ADC values alone. In the peritumoral areas of untreated high-grade gliomas and metastases, the mean rCBV values were higher for high-grade gliomas (1.7+/-0.37) than for metastases (0.54+/-0.18) while the mean ADC values were higher for metastases. The rCBV values of four lymphomas were low and the signal intensity-time curves revealed a significant increase in signal intensity after the first pass of gadobenate dimeglumine. Diffusion and perfusion imaging, even with relatively short imaging and data processing times, provide important information for lesion characterization. PMID- 16470376 TI - Extensive sharing of MHC class II alleles between rhesus and cynomolgus macaques. AB - In contrast to rhesus monkeys, substantial knowledge on cynomolgus monkey major histocompatibility complex (MHC) class II haplotypes is lacking. Therefore, 17 animals, including one pedigreed family, were thoroughly characterized for polymorphic Mhc class II region genes as well as their mitochondrial DNA (mtDNA) sequences. Different cynomolgus macaque populations appear to exhibit unique mtDNA profiles reflecting their geographic origin. Within the present panel, 10 Mafa-DPB1, 14 Mafa-DQA1, 12 Mafa-DQB1, and 35 Mafa-DRB exon 2 sequences were identified. All of these alleles cluster into lineages that were previously described for rhesus macaques. Moreover, about half of the Mafa-DPB1, Mafa-DQA1, and Mafa-DQB1 alleles and one third of the Mafa-DRB exon 2 sequences are identical to rhesus macaque orthologues. Such a high level of Mhc class II allele sharing has not been reported for primate species. Pedigree analysis allowed the characterization of nine distinct Mafa class II haplotypes, and seven additional ones could be deduced. Two of these haplotypes harbor a duplication of the Mafa DQB1 locus. Despite extensive allele sharing, rhesus and cynomolgus monkeys do not appear to possess identical Mhc class II haplotypes, thus illustrating that new haplotypes were generated after speciation by recombination-like processes. PMID- 16470377 TI - NKG2D splice variants: a reexamination of adaptor molecule associations. AB - NKG2D is a homodimeric C-type lectin-related receptor expressed on natural killer (NK) cells and T cells. In mice, alternative deoxyribonucleic acid (DNA) splicing generates two isoforms of NKG2D that differ in the length of their cytoplasmic domains. Their ability to induce cellular activation is mediated via association with two membrane-bound, signaling adaptor molecules, DAP10 and DAP12. It has been reported that the long form of NKG2D associates exclusively with DAP10, whereas the short variant can interact with either adaptor. The short isoform was reported to be almost undetectable in naive NK cells. Using two distinct cell types, we demonstrate that like the short isoform, the long variant of NKG2D also associates not only with DAP10 but also with DAP12. Using reporter cells (70Z/3), we demonstrate that DAP12 can compete equally with DAP10 for association with both variants of NKG2D when DAP10 and DAP12 are coexpressed. Cross-linking either isoform of NKG2D induces a calcium flux when associated exclusively with DAP10 or DAP12. Moreover, using quantitative polymerase chain reaction (PCR), we also show that the short isoform of NKG2D is expressed in naive NK cells. Our data suggest that signaling via mouse NKG2D isoforms is more complex than originally presented. PMID- 16470378 TI - Actin and amphiphilic polymers influence on channel formation by Syringomycin E in lipid bilayers. AB - The bacterial lipodepsipeptide syringomycin E (SRE) added to one (cis-) side of bilayer lipid membrane forms voltage dependent ion channels. It was found that G actin increased the SRE-induced membrane conductance due to formation of additional SRE-channels only in the case when actin and SRE were applied to opposite sides of a lipid bilayer. The time course of conductance relaxation depended on the sequence of SRE and actin addition, suggesting that actin binds to the lipid bilayer and binding is a limiting step for SRE-channel formation. G actin adsorption on the membrane was irreversible. The amphiphilic polymers, Konig's polyanion (KP) and poly(Lys, Trp) (PLT) produced the actin-like effect. It was shown that the increase in the SRE membrane activity was due to hydrophobic interactions between the adsorbing molecules and membrane. Nevertheless, hydrophobic interactions were not sufficient for the increase of SRE channel-forming activity. The dependence of the number of SRE-channels on the concentration of adsorbing species gave an S-shaped curve indicating cooperative adsorption of the species. Kinetic analysis of SRE-channel number growth led to the conclusion that the actin, KP, and PLT molecules form aggregates (domains) on the trans-monolayer. It is suggested that an excess of SRE-channel formation occurs within the regions of the cis-monolayer adjacent to the domains of the adsorbed molecules, which increase the effective concentration of SRE-channel precursors. PMID- 16470379 TI - [Protrusio bulbi]. PMID- 16470380 TI - [Diagnostic procedure for uveitis patients: reduction of costs by a targeted assessment of laboratory tests based on clinical findings]. AB - BACKGROUND: Interdisciplinarity is an essential prerequisite for extensive and high-quality diagnostic work-up, therapy, and care of patients with inflammatory rheumatic disease. The ophthalmological status significantly narrows down the possible causes for the primary disease and facilitates an effective diagnostic approach. METHODS: We developed an assessment procedure according to the individual clinical findings to use as an ideal diagnostic strategy for an interdisciplinary center. To permit comparability we first introduced this new diagnostic tool in November 2002 (intervention), replacing the existing cost intensive standard diagnostic procedure for uveitis patients. RESULTS: The average costs for laboratory tests (calculated according to GOA numbers, scale of fees for physicians) were reduced from 670+/-11 euros per patient before the intervention to 238+/-18 euros per patient after the intervention. Before intervention 16 of 31 (52%) could be assigned to a specified disease compared to 27 of 53 (51%) after the intervention. With our strategy we were able to obtain a 64% reduction of costs in the diagnostics of uveitis patients without a deterioration of our diagnostic quality. CONCLUSION: It is possible to reduce costs and still improve the care of uveitis patients by using a targeted assessment according to the individual clinical findings. PMID- 16470381 TI - [Optical lamellar-penetrating keratoplasty with stem cell transplantation in high risk cases]. AB - BACKGROUND: Stem cells of the corneal epithelium are located mainly at the sclerocorneal limbus, and are essential for the maintenance of a healthy corneal surface. Limbal stem cell deficiency leads--depending on intensity and extension- to several corneal alterations. The appearance of limbal stem cell deficiency is chiefly concentrated on pemphigoids, Herpes relapses and alkali burns and represents the main part of high-risk keratoplasty. The different types of limbal stem cell deficiency (partial or total) are usually treated with several surgical techniques. Total limbal deficiency can be treated with limbal transplantation, either of the healthy eye (limbal autograft), or using material of another donor (limbal allograft). METHODS AND RESULTS: Lamellar-penetrating keratoplasty (L-P KP) has been carried out in five patients. Unlike recent surgical techniques containing ring-shaped or circular transplantations, a stripe-like lamellar preparation was performed. L-P-KP involves a peripheral lamellar and a central penetrating keratoplasty combined with limbal stem cell transplantation. In four of six cases, the transplant has been successfully kept clear without the occurrence of corneal decompensation or ulceration. No major changes could be seen between pre- and postoperative visual acuity. The mean observation period was 24 months. DISCUSSION: L-P-KP offers a new surgical opportunity in the treatment of high-risk cases of limbal deficiency. PMID- 16470383 TI - Enhanced gene targeting frequency in ku70 and ku80 disruption mutants of Aspergillus sojae and Aspergillus oryzae. AB - In the koji molds Aspergillus sojae and Aspergillus oryzae, exogenous DNA is integrated in the genome, in most cases irrespective of the sequence homology, suggesting that DNA integration occurs predominantly through a nonhomologous end joining pathway where two ku genes, namely, ku70 and ku80, play a key role. To determine the effect of ku gene disruption on the gene targeting frequency, we constructed ku70-, ku80-, and ku70-ku80-disrupted strains of A. sojae and A. oryzae. The gene targeting frequency of the tannase gene in ku70 and ku80 strains of both Aspergillus species was markedly enhanced as compared with that of the parental strains. The gene targeting frequency of the aflR and ku80 genes was also enhanced in an A. sojae ku70 background. Therefore, the koji mold strains with ku-disrupted genes will be excellent tools as hosts for efficient gene targeting. PMID- 16470382 TI - Characterization of a novel prospero-related homeobox gene, Prox2. AB - Prospero-related homeobox genes have been identified from various multi-cellular organisms and play important roles in development as a cell fate determinant. Mouse Prox1 is essential for embryogenesis and is required to differentiate horizontal cells in the retina. Here we describe a novel prospero family member, Prox2. Transcriptional reporter assays demonstrated that mouse Prox2 is a transcriptional activator and the N-terminal region has been identified as an activation domain. The expression of mouse Prox2 was detected in postnatal eyes and adult testes as well as embryos. To investigate the in vivo role of Prox2, we generated the Prox2 mutant allele, Prox2-, by homologous recombination in mouse ES cells. Prox2- lacks the first coding exon that encodes a translational start site and a part of homeodomain. In spite of the Prox2 expression during embryogenesis, Prox2- homozygous mutant mice were born at the expected Mendelian ratio without overt abnormalities. Histological analyses revealed that Prox2- homozygous eyes retained the organized layer structure including three nuclear layers and differentiated horizontal cells. Prox2- homozygous mutant males produced elongated spermatids and were fertile. These results demonstrate that mouse Prox2 is dispensable for embryonic development, horizontal cell generation and fertility in contrast to mouse Prox1. PMID- 16470384 TI - Direct interaction of Saccharomyces cerevisiae Faa1p with the Omi/HtrA protease orthologue Ynm3p alters lipid homeostasis. AB - In yeast, long chain acyl-CoA synthetase (ACSL) activity is required for fatty acid uptake, metabolism and fatty acid-dependent transcriptional control. The major ACSL contributing these functions is Faa1p. In a yeast two-hybrid screen, the Omi/HtrA serine protease family orthologue Ynm3p (YNL123w) was identified as a specific interactor with Faa1p. Interaction of Ynm3p and Faa1p was confirmed by co-immunoprecipitation. Disruption of the YNM3 gene encoding Ynm3p resulted in increased fatty acid uptake, triglyceride accumulation and reduced expression of the fatty acid-responsive OLE1 gene encoding the essential Delta(9)-acyl-CoA desaturase. These changes were linked with increased Faa1p and Faa4p ACSL activities. We propose that Ynm3p modulates fatty acid metabolism and gene regulation through negative regulation of ACSL activity. Additional strain specific phenotypes associated with deletion of YNM3 included inability to grow on non-fermentable carbon sources and altered cellular morphology. PMID- 16470385 TI - Neuromuscular performance and knee laxity do not change across the menstrual cycle in female athletes. AB - Female athletes incur anterior cruciate ligament ruptures at a rate at least twice that of male athletes. Hypothesized factors for the increased injury risk in females include biomechanical, neuromuscular, and hormonal differences between genders. A wealth of literature exists examining these potential predispositions individually, but the interactions between these factors have not been examined extensively. Our purpose was to investigate changes in neuromuscular control and laxity at the knee across the menstrual cycle of healthy females. Fourteen female collegiate athletes with normal, documented ovulatory menstrual cycles, confirmed ovulation, and no history of serious knee injury participated. The presence and timing of ovulation was determined during a screening cycle with ovulation detection kits and during an experimental cycle with collection of daily urine samples and subsequent analysis of urinary estrone-3-glucuronide (E3G) and pregnanediol-3-glucoronide (PdG), which correlate with circulating estrogen and progesterone. Each subject had measures of knee neuromuscular performance and laxity once during the mid-follicular, ovulatory, and mid-luteal stages of her menstrual cycle. The test battery included assessments of knee flexion and extension peak torque, passive knee joint position sense, and postural control in single leg stance. Knee joint laxity was measured with an arthrometer. Analyses of variance revealed that E3G and PdG levels were significantly different across the three testing sessions, but there were no significant differences in the measures of strength, joint position sense, postural control, or laxity. No significant correlations were found between changes in E3G or PdG levels and changes in the performance and laxity measures between sessions. These results suggest that neuromuscular control and knee joint laxity do not change substantially across the menstrual cycle of females despite varying estrogen and progesterone levels. PMID- 16470386 TI - Transverse femoral fixation in anterior cruciate ligament (ACL) reconstruction with hamstrings grafts: an anatomic study about the relationships between the transcondylar device and the posterolateral structures of the knee. AB - Iatrogenic risks concerning femoral cross pin devices for anterior criciate ligament (ACL) reconstruction using hamstrings grafts have not been evaluated. The aim of the study was to state anatomic relationships between the epicondylar cross pin hole and the posterolateral attachments of the knee, and to settle safe zones and technical improvements for its use. It is a descriptive anatomical study. Dissections were performed and measurements taken on 20 (2x10) fresh frozen cadaveric knees after standard arthroscopic and femoral ACL reconstruction procedure using the Transfix cross pin femoral device (Arthrex, Naples, FL, USA). In group I, femoral tunnel was performed through the tibial tunnel. In group II, an anteromedial portal technique was used. Group I: no iatrogenic injury. The cross pin was at 36.5+/-2.8 mm (33-41) from the femoral inferior articular area, and at 12.3+/-3.2 mm (7-17) from the lateral collateral ligament (LCL) insertion. Group II: in a half cases, there was an injury of the LCL, close to its insertion. The cross pin was at an average of 5.6+/-5.4 mm (0-15) from the LCL, and significantly closer in group II than in group I (P<0.01). In group I, there is a high correlation between cross pin to LCL distance and knee flexion obtained to drill the femoral tunnel (r (2)=0.75, P<0.01). In conclusion, there is a specific iatrogenic risk of transverse femoral fixation using the anteromedial portal approach, whereas the transtibial approach seems to be safe. Nevertheless, it can decrease with a correct femoral tunnel depth and an operative knee flexion over 130 degrees , in order to move LCL insertion and posterior cortical wall away. PMID- 16470388 TI - Invertebrate studies and their ongoing contributions to neuroscience. AB - Invertebrates have been deployed very successfully in experimental studies of the nervous system and neuromuscular junctions. Many important discoveries on axonal conduction, synaptic transmission, integrative neurobiology and behaviour have been made by investigations of these remarkable animals. Their advantages as model organisms for investigations of nervous systems include (a) the large diameter of neurons, glia and muscle cells of some invertebrates, thereby facilitating microelectrode recordings; (b) simple nervous systems with few neurons, enhancing the tractability of neuronal circuitry; and (c) well-defined behaviours, which lend themselves to physiological and genetic dissection. Genetic model organisms such as Drosophila melanogaster and Caenorhabditis elegans have provided powerful genetic approaches to central questions concerning nervous system development, learning and memory and the cellular and molecular basis of behaviour. The process of attributing function to particular gene products has been greatly accelerated in recent years with access to entire genome sequences and the application of reverse genetic (e.g. RNA interference, RNAi) and other post-genome technologies (e.g. microarrays). Studies of many other invertebrates, notably the honeybee (Apis mellifera), a nudibranch mollusc (Aplysia californica), locusts, lobsters, crabs, annelids and jellyfish have all assisted in the development of major concepts in neuroscience. The future is equally bright with ease of access to genome-wide reverse genetic technologies, and the development of optical recordings using voltage and intracellular calcium sensors genetically targeted to selected individual and groups of neurons. PMID- 16470390 TI - Anatomical hemispherectomy for intractable seizures: excellent seizure control, low morbidity and no superficial cerebral haemosiderosis. AB - OBJECTIVE: This current study was performed to evaluate whether superficial cerebral haemosiderosis (SCH) is still a complication of modern day anatomical hemispherectomy. METHODS: We report a 13-year institutional experience with anatomical hemispherectomy for intractable epilepsy. Seizure control at a mean follow-up interval of 7 years was 83%. Though one patient died post-operatively from a non-neurosurgical complication, mortality was otherwise zero and morbidity minimal. The much-described complication of SCH following anatomical hemispherectomy was non-existent. We explain the history of SCH as a complication of anatomical hemispherectomy, and the measures that are presently taken to prevent it. CONCLUSIONS: We suggest that the importance of SCH in modern epilepsy surgery is probably over-emphasised. PMID- 16470387 TI - Development, maturation and subsequent activation of follicular dendritic cells (FDC): immunohistochemical observation of human fetal and adult lymph nodes. AB - To elucidate the processes involved in development and activation of human follicular dendritic cells (FDC), immunohistochemistry was performed on paraffin sections of fetal lymph nodes (FLN) obtained from archived autopsy material, and of adult reactive lymph nodes (ARLNs) excised for diagnostic purpose, using a panel of antibodies. Our study showed that tiny clusters of CNA.42(+ )KiM4p(+) cells, surrounded by some B-lymphocytes, initially arose in the cortical area of underdeveloped FLN around the 20th gestational week. No co-expression of CD21 and CD35 was found. In the relatively developed FLN of the same gestational age, small eddies of immature FDC, which expressed CD21, CD35, and nerve growth factor receptor (NGFR), as well as CNA.42 and KiM4p, were observed within ill-defined aggregations of B-lymphocytes. As gestation progressed, more B-lymphocytes assembled in a compact manner and formed primary lymphoid follicles containing an extending web of mature FDC, which expressed CNA.42, KiM4p, CD21, CD35, NGFR, and sometimes CD23 and X-11. In well-developed secondary follicles of ARLNs, activated FDC expressed additional molecules such as CD55, CD106, and S100alpha. Our observations identified the processes of phenotypic alteration of human FDC and established practical indicators determining their developmental stage and functional phase. PMID- 16470391 TI - Bladder lithiasis: from open surgery to lithotripsy. AB - Bladder calculi account for 5% of urinary calculi and usually occur because of bladder outlet obstruction, neurogenic voiding dysfunction, infection, or foreign bodies. Children remain at high risk for developing bladder lithiasis in endemic areas. Males with prostate disease or relevant surgery and women who undergo anti incontinence surgery are at a higher risk for developing vesical lithiasis. Open surgery remains the main treatment of bladder calculus in children. In adults, the classical treatment for bladder calculi is endoscopic transurethral disintegration with mechanical cystolithotripsy, ultrasound, electrohydraulic lithotripsy, Swiss Lithoclast, and holmium:YAG laser. Novel modifications of these treatment modalities have been used for large calculi. Open and endoscopic surgery requires anesthesia and hospitalization. Alternatively, extracorporeal shock wave lithotripsy has been demonstrated to be simple, effective, and well tolerated in high-risk patients. Recently, simultaneous percutaneous suprapubic and transurethral cystolithotripsy has been tested as well as percutaneous cystolithotomy by using a laparoscopic entrapment sac. PMID- 16470392 TI - Familial expansile osteolysis--not exclusively an adult disorder. AB - Familial expansile osteolysis (FEO, MIM174810) is a rare syndrome which was observed world-wide in only three kinships and in two unrelated American individuals. We report a patient with familial expansile osteolysis from the Czech Republic, not related to the previously reported cases. This patient's extraordinary clinical course does not conform to the ordinary. Her radiographic bone involvement was unusually extensive, involving most of the peripheral skeleton and the skull. This case documents that familial expansile osteolysis is not only a disease of adults but does occur in childhood. PMID- 16470393 TI - Migrating lumbar facet joint cysts. AB - OBJECTIVE: The majority of lumbar facet joint cysts (LFJCs) are located in the spinal canal, on the medial aspect of the facet joint with characteristic diagnostic features. When they migrate away from the joint of origin, they cause diagnostic problems. MATERIALS AND METHODS: In a 7-year period we examined by computed tomography (CT) and magnetic resonance (MR) imaging five unusual cases of facet joint cysts which migrated from the facet joint of origin. RESULTS: Three LFJCs were identified in the right S1 foramen, one in the right L5-S1 neural foramen and one in the left erector spinae and multifidus muscles between the levels of L2-L4 spinous process. CONCLUSIONS: Awareness that spinal lesions identified at MRI and CT could be due to migrating facet joint cyst requires a high level of suspicion. The identification of the appositional contact of the cyst and the facet joint needs to be actively sought in the presence of degenerative facet joints. PMID- 16470394 TI - Assessment of enthesitis in ankylosing spondylitis by power Doppler ultrasonography. AB - OBJECTIVE: To evaluate the relationship between power Doppler ultrasonography (PDUS) assessment and clinical variables including enthesitis index, pain threshold and disease activity parameters, and to document grey-scale US findings of the 13 entheses examined. DESIGN AND PATIENTS: A total of 390 entheses were examined in thirty patients with AS, and clinical variables of the Maastricht Ankylosing Spondylitis Enthesitis Index (MASES), anthropometric measurements, disease activity and functional parameters were documented. A total MASES score by palpation (t-PS) and algometric pressure pain threshold (t-PPT) was obtained. Grey scale and PDUS examination of 13 entheses were performed. Grey-scale changes such as altered tendon echogenity, calcification, cortical reactive changes and bursitis were noted, and flow on PDUS was graded semi-quantitatively. RESULTS: Cumulative power Doppler (t-PDS) score significantly correlated with t-PS and t PPT. Ultimate correlations were found between power Doppler scores and pain, disease activity and disability parameters. Changes in grey scale combined with PDUS were more prevalent in lower-extremity entheses. The intraobserver agreement of flow signal grading was excellent (kappa = 0.82). Clinical and sonographic results were concordant for three regions, but were discordant for four regions where tenderness was accepted as the sole clinical manifestation of enthesis. CONCLUSION: Pain or tenderness is associated with increased vascularity of entheses. Power Doppler US examination of the entheses may be useful and complementary to the clinical evaluation, and further research is needed to assess its role in diagnosis and follow-up of disease course. PMID- 16470395 TI - Edematous Schmorl's nodes on thoracolumbar MR imaging: characteristic patterns and changes over time. AB - OBJECTIVE: To describe the patterns and note the evolution of edematous Schmorl's nodes. MATERIALS AND METHODS: In 47 patients (M:F=26:21, 24-86 years, average 60), 84 Schmorls nodes with T2 hyperintensity with serial MR exams were evaluated. Interval between MR exams was 2-72 months (average 17). Two observers noted size, location, margins, internal and surrounding T1/T2 signal, adjacent disc herniation or bulge, concentric ring, underlying fracture, malignancy, infection, or prior disc surgery, and serial MR changes in these characteristics over time. RESULTS: Node size averaged 7x9 mm. Most were located at L3 (29%, 24/84), L4 (19%, 16/84) and L2 (13%, 11/84), at the central (39%, 33/84) or outer (30%, 25/84) third of the endplate. 55% (39/71) had a bulging disc, 7% (5/71) had disc herniation. 10% (8/84) had evidence of associated fracture, 17% (14/84) tumor, 7% (6/84) infection. Most nodes had well-defined margins (82%, 69/84). The most common node internal signal was isointense to adjacent disc on T1/T2 (33%, 28/84); surrounding marrow was most commonly hypointense on T1 and hyperintense on T2 (54%, 38/71). A common finding was concentric rings (38%, 32/84) in the marrow surrounding the node, a finding which had 72% negative predictive value for absence of infection, tumor and fracture. On follow-up, there was no interval change in node size in 46%(39/84) of Schmorl's nodes. 26% (22/84) had increased size. Most (60%, 50/84) showed no temporal change in internal T2 signal. 21% (18/84) of nodes showed decreased internal T2 signal, 13% (11/84) showed increased T2 signal. Regarding the surrounding marrow, most (58%, 49/84) showed no temporal change in T2 signal; 21%(18/84) showed decreased T2 signal, 13% (11/84) showed increased T2 signal. In 13 Schmorl's nodes with intranodal enhancement, eight (62%) showed no interval change; among eight with enhancement in surrounding marrow, five (63%) showed no change on follow-up. CONCLUSION: Although most remain unchanged, a relatively large minority of edematous Schmorl's nodes evolve in size and signal over a relatively short time. Some evolve to form well-defined concentric rings in the surrounding marrow that appear to be analogous to degenerative changes of endplates. Concentric ring formation has a high negative predictive value for "idiopathic" Schmorl's nodes without underlying fracture, infection, or malignancy. PMID- 16470396 TI - Fractures of the proximal femur: correlates of radiological evidence of osteoporosis. AB - OBJECTIVE: Fractures of the proximal femur are common sequelae of osteoporosis, and are responsible for significant morbidity and mortality in elderly patients worldwide. Plain film radiographic assessment methods to assess for fracture risk may be of particular value. DESIGN AND PATIENTS: The authors present the results of biomechanical testing, radiographic imaging, and histologic exam of 20 embalmed human bone specimens, with implications for clinical correlation of radiologic findings. Authors assessed bone architecture using the Singh Index, using a blinded 3-rater system to reduce bias and measure intra-observer reliability. After loading to failure with ultimate tensile strength (UTS), bone specimens were assessed by fracture location type and by trabecular bone volume (TBV). RESULTS: Singh scoring was performed with Inter-Class Correlation of 0.80 (F=0.24, by ICC Portney Model 2). A statistically-significant difference among the UTS distributions was noted for UTS by Fracture Site (F=4.49, p=0.026, by ANOVA). No significant association of Singh Index with TBV, or TBV with UTS, was observed, although a trend toward greater UTS with higher Singh grade was observed. CONCLUSIONS: The authors propose that the Singh Index is a valuable and reliable indicator which may reflect structural integrity in trabecular bone. Fracture site along the femur is associated with tensile strength. The authors, in the light of these findings, address the promise and potential impact of prophylactic hip augmentation in populations at risk for femoral neck pathology. PMID- 16470397 TI - Evaluation of a Dallas Pain Questionnaire classification in relation to outcome in lumbar spinal fusion. AB - Several studies have suggested that psychosocial distress is an important predictor of negative outcome in lumbar spine surgery. Ozguler et al. [Spine 27:1783-1789, 2002, 18] described a classification tool for low back pain patients using the Dallas Pain Questionnaire (DPQ) which included a measure of distress. We wanted to evaluate the ability of this classification tool to predict the outcome in spinal fusion patients. Five hundred and sixty-six patients (239 men, 327 women; mean age 46, range 18-81) operated between 1992 and 2002, with a complete DPQ preoperatively and after a minimum of 1-year follow-up, were included. They were classified preoperatively and at follow-up into four groups: group 1 (slight disability), group 2 (intermediate disability), group 3 (major disability) and group 4 (major disability and emotional distress). Using logistic regression, seven predictor variables were investigated: age (-39 years/40-59 years/60+ years), Gender (male/female), Indication (spondylolisthesis/primary degeneration/secondary degeneration), Work status (working/without work or on sick leave/retired or pensioned), Duration of pain (less than 1 year/1-2 years/more than 2 years), Presence of radiating pain (yes/no) and disability/distress [intermediate disability (group 1-2)/major disability (group 3)/major disability and distress (group 4)]. The outcome variable was disability at follow-up (low = group1 + 2/high = group 3 + 4). Preoperative classification was group 1, 1%; group 2, 14%; group 3, 37%; group 4, 48%. Variables found to predict high disability at follow-up were secondary degeneration Odds Ratio (OR) 1.61 (P=0.020), being retired/pensioned OR 3.48 (P<0.0005), age between 40-59 years OR 1.68 (P=0.011), belonging to group 3 OR 2.69 (P=0.003) or belonging to group 4 OR 5.53 (P<0.0005). The classification based on the DPQ were able to identify lumbar spinal fusion patients with a considerable amount of psychological distress in their symptomatology. Furthermore, the presence of distress, as determined by this classification, was a highly significant risk factor for inferior outcome. PMID- 16470398 TI - Comparison of anterior and posterior double-rod instrumentation for thoracic idiopathic scoliosis: results of 141 patients. AB - Ventral derotation spondylodesis, according to Zielke, achieves good results in operative treatment of idiopathic thoracic scolioses. Corrections of scoliotic major and secondary curve as well as derotation of the spine are reliably performed. The high rate of rod fractures with subsequent correction loss as well as a proportionate kyphogenic effect represents a problem. By keeping to the correcting principle, anterior double-rod instrumentation (Halm-Zielke Instrumentation) is to be stable in a similar way as posterior double-rod systems. Thus, it is done to facilitate brace-free postoperative care and to prevent excessive kyphotic pattern of the spine. In this prospective study, we retrospectively collected data. We performed radiological follow-up of two groups of patients with idiopathic thoracic scoliosis (King II, III and IV) undergoing an operation with posterior approach (USS instrumentation, posterior group, n=104) in 1997 and 1998 or being corrected with an anterior fusion (Halm-Zielke instrumentation, anterior group, n=37) between 2000 and 2001. Mean age of all patients for operation was 15+/-4 years. Follow-up was performed after 4+/-2 years on average. Preoperative measurements of the major and secondary curve, the lateral profile, rotation and frontal balance (C7 to S1) did not show any significant differences apart from a more severe scoliotic curve in the lumbar spine for the anterior group with appropriately higher lumbar rotation. During follow-up we noticed similar corrections of the thoracic major and lumbar curve in both groups ranging from 49 to 56%. In case of hypokyphotic (T4-T12or=40 degrees ) scolioses were flattened by posterior spinal fusion; the effect of anterior spinal fusion was not significant. Correction of thoracic and lumbar rotation in the anterior group by 37 or 30% was more significant than in the posterior group by 27 or 20%. There was no impact of anterior technique on the balance of the spine whereas the latter shifted by an average of 7 mm to the left in the posterior group. The number of fused segments was significantly smaller in the anterior group with 7+/ 1 vertebral bodies (posterior, 11+/-1 vertebral bodies). Rates of complication were identical with 11 or 12% in both groups during follow-up. Anterior and posterior double-rod instrumentations result in comparable corrections for idiopathic thoracic scoliosis of the major and secondary curve. In case of posterior technique, however, four vertebral bodies less were integrated in spondylodesis on average. Balance of the spine did not change after anterior spondylodesis; however, it declined by using the posterior technique. Augmentation of the anterior threaded rod combined with a solid second rod significantly decreases the rate of implant breakages and reliably reduces consecutive correction losses. PMID- 16470399 TI - Square-lashing technique in segmental spinal instrumentation: a biomechanical study. AB - Sublaminar wires have been used for many years for segmental spinal instrumentation in scoliosis surgery. More recently, stainless steel wires have been replaced by titanium cables. However, in rigid scoliotic curves, sublaminar wires or simple cables can either brake or pull out. The square-lashing technique was devised to avoid complications such as cable breakage or lamina cutout. The purpose of the study was therefore to test biomechanically the pull out and failure mode of simple sublaminar constructs versus the square-lashing technique. Individual vertebrae were subjected to pullout testing having one of two different constructs (single loop and square lashing) using either monofilament wire or multifilament cables. Four different methods of fixation were therefore tested: single wire construct, square-lashing wiring construct, single cable construct, and square-lashing cable construct. Ultimate failure load and failure mechanism were recorded. For the single wire the construct failed 12/16 times by wire breakage with an average ultimate failure load of 793 N. For the square lashing wire the construct failed with pedicle fracture in 14/16, one bilateral lamina fracture, and one wire breakage. Ultimate failure load average was 1,239 N For the single cable the construct failed 12/16 times due to cable breakage (average force 1,162 N). 10/12 of these breakages were where the cable looped over the rod. For the square-lashing cable all of these constructs (16/16) failed by fracture of the pedicle with an average ultimate failure load of 1,388 N. The square-lashing construct had a higher pullout strength than the single loop and almost no cutting out from the lamina. The square-lashing technique with cables may therefore represent a new advance in segmental spinal instrumentation. PMID- 16470400 TI - Previous experience of ethanol withdrawal increases withdrawal-induced c-fos expression in limbic areas, but not withdrawal-induced anxiety and prevents withdrawal-induced elevations in plasma corticosterone. AB - RATIONALE: Increased anxiety is a characteristic of the acute ethanol withdrawal syndrome. Repeated exposure of rats to withdrawal from chronic ethanol increases sensitivity to seizures. OBJECTIVES: We investigated whether repeated withdrawal experience increases withdrawal-induced anxiety and stress, and if it changes withdrawal-induced activation of related brain areas. METHODS: Rats were chronically treated with an ethanol-containing liquid diet either for 24 days continuously (single withdrawal, SWD) or interspersed with 2x3-day withdrawal periods (repeated withdrawal, RWD), or with a control diet. Eight hours after ethanol withdrawal, anxiety-like behaviour was tested in the elevated plus-maze, blood corticosterone levels were measured, and expression level of markers of neuronal activity and plasticity, c-fos and zif268, was assessed. RESULTS: Eight hours after ethanol withdrawal, SWD rats showed increased anxiety on the elevated plus-maze relative to control rats. Rats given previous withdrawal experiences did not show further increases in measures of anxiety. Corticosterone levels were elevated during withdrawal in SWD rats but not in RWD rats. RWD resulted in marked increases in c-fos expression in amygdala, hippocampus, nucleus accumbens and dorsolateral periaqueductal grey. In contrast, zif268 expression was not increased after RWD, and in central amygdala the marked increase in zif268 seen after SWD was absent after RWD. CONCLUSIONS: The data suggest increased ability of withdrawal to activate neuronal circuits but reduced plasticity after RWD. We suggest parallels between the consequences of repeated ethanol withdrawal and repeated exposure to stress, and discuss implications of withdrawal for brain plasticity. PMID- 16470401 TI - In vitro and in vivo studies in rats with LY293558 suggest AMPA/kainate receptor blockade as a novel potential mechanism for the therapeutic treatment of anxiety disorders. AB - RATIONALE: Although convergent evidence exists for a role of glutamate in the regulation of anxiety, the involvement of specific glutamate receptor subtypes has yet to be defined. OBJECTIVE: To evaluate the potential for blockade of alpha amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA)/kainate receptors to produce anxioltyic-like effects with the AMPA/GLU(K5) (kainate) antagonist (3S,4aR,6R,8aR)-6-[2-(1(2)H-tetrazole-5yl)ethyl]decahydroisoquinoline-3carboxylic acid (LY293558) MATERIALS AND METHODS: Punished responding of rats was used to determine the efficacy of LY293558. Other in vivo and in vitro studies further characterized the specificity of LY293558 for AMPA/kainate receptors. RESULTS: LY293558 had a rank order of potency of GLU(K5) > or = GLU(K5/6) approximately GLU(A2i) approximately GLU(K2/5) approximately GLU(A1i) approximately GLU(A2o) approximately GLU(A3i) approximately GLU(A1o) > or = GLU(A3o) > or = GLU(A4i) approximately GLU(A4o) and >100 microM affinity for rat cortical GABA(A) receptors. Comparison of the blockade of AMPA- vs N-methyl-D-aspartate (NMDA) induced inward currents demonstrated that LY293558 was five-fold more potent as an antagonist at AMPA vs NMDA receptors in vitro. In keeping with the low affinity of LY293558 for NMDA receptors, LY293558 was not effective in preventing NMDA-induced seizures in mice. LY293558 increased punished responding, a preclinical predictor of anxiolytic efficacy, at a dose that decreased unpunished responding (10 mg/kg, i.p.). Chlordiazepoxide produced comparable increases in both punished and unpunished responding. The NMDA antagonist dizocilpine [(+)-MK 801] also increased both punished and unpunished responding. CONCLUSIONS: These data along with those in the literature suggest that AMPA and/or kainate receptor blockade may be an important component to producing anxiolytic-like effects and may therefore be a target for compounds with efficacy in the therapeutic treatment of anxiety disorders. PMID- 16470402 TI - Withdrawal from continuous amphetamine administration abolishes latent inhibition but leaves prepulse inhibition intact. AB - RATIONALE: Schizophrenia has been associated with dysregulation of dopamine (DA) transmission and impairment in a number of experimental tasks, including sensorimotor gating assessed using prepulse inhibition (PPI) and selective attention assessed using latent inhibition (LI). We have demonstrated in previous studies that after withdrawal from escalating (ESC) dosages of amphetamine (AMPH), animals exhibited disruption of LI but no alteration of PPI. Moreover, these animals always showed behavioural sensitization to an AMPH challenge. OBJECTIVE: In this study, we were interested in testing whether a different administration schedule would elicit disruption of both LI and PPI. METHODS: Animals were treated with continuous AMPH release (via osmotic mini-pumps at a dosage of 10 mg kg(-1) day(-1) for 7 days) and tested for their performance in L and PPI during withdrawal in a drug free state. Rats received AMPH treatment during the induction phase in their home cages or in the activity chambers. Following withdrawal, the expression of behavioural sensitization to an AMPH challenge was tested in both cases in the activity chambers. RESULTS: Animals pretreated with AMPH from both groups did not exhibit behavioural sensitization. Withdrawal from continuous administration induced LI attenuation with no effect on PPI. CONCLUSIONS: These findings are similar to what was previously found with respect to an ESC AMPH regime. The only difference between the schedules was that the ESC AMPH schedule led to behavioural sensitization whereas the continuous AMPH did not. It is suggested that the expression of sensitization may not be a prerequisite for observed LI disruption. PMID- 16470403 TI - Absence of quasi-morphine withdrawal syndrome in adenosine A2A receptor knockout mice. AB - RATIONALE: Caffeine and other methylxanthines induce behavioral activation and anxiety responses in mice via antagonist action at A2A adenosine receptors. When combined with the opioid antagonist naloxone, methylxanthines produce a characteristic quasi-morphine withdrawal syndrome (QMWS) in opiate-naive animals. OBJECTIVES: The aim of this study was to establish the role of A2A receptors in the quasi-morphine withdrawal syndrome induced by co-administration of caffeine and naloxone and in the behavioral effects of caffeine. METHODS: We have used A2A receptor knockout (A(2A)R(-/-)) mice in comparison with their wild-type and heterozygous littermates to measure locomotor activity in the open field and withdrawal symptoms induced by caffeine and naloxone. Naive wild-type and knockout mice were also examined for enkephalin and dynorphin mRNA expression by in situ hybridization and for mu-opiate receptor by ligand binding autoradiography to check for possible opiate receptor changes induced by A2A receptor inactivation. RESULTS: Caffeine increases locomotion and anxiety in wild type animals, but it has no psychomotor effects in A(2A)R(-/-) mice. Co administration of caffeine (20 mg/kg) and naloxone (2 mg/kg) resulted in a severe quasi-morphine withdrawal syndrome in wild-type mice that was almost completely abolished in A(2A)R(-/-) mice. Heterozygous animals exhibited a 40% reduction in withdrawal symptoms, suggesting that there is no genetic/developmental compensation for the inactivation of one of the A(2A)R alleles. A(2A)R(-/-) and wild-type mice have similar levels of striatal mu-opioid receptors, thus the effect is not due to altered opioid receptor expression. CONCLUSIONS: Our results demonstrate that A2A receptors are required for the induction of quasi-morphine withdrawal syndrome by co-administration of caffeine and naloxone and implicate striatal A2A receptors and mu-opiate receptors in tonic inhibition of motor activity in the striatum. PMID- 16470405 TI - Serotonin 1B and 2C receptor interactions in the modulation of feeding behaviour in the mouse. AB - RATIONALE: To examine the functional relationship between 5-HT1B receptors (5 HT1B-R) and 5-HT2C receptors (5-HT2C-R) in the control of food intake. OBJECTIVES: To compare the hypophagic effect of the 5-HT(2C/1B)-R agonist m chlorophenylpiperazine (mCPP), with that of the selective 5-HT1B-R agonist CP 94,253 in both wildtype (WT) and 5-HT2C knockout (KO) mice. METHODS: The hypophagic effects of mCPP (1, 3 and 5.6 mg/kg) and CP-94,253 (5, 10 and 20 mg/kg) were assessed in WT and 5-HT2C KO mice using the behavioural satiety sequence paradigm. The effects of pre-treatment with the selective 5-HT2C-R antagonist SB 242,084 (0.5 and 1.5 mg/kg) were assessed in two groups of WT mice, with each group given only mCPP or CP-94,253. RESULTS: The 5-HT(2C/1B) receptor agonist mCPP and the selective 5-HT1B receptor agonist CP-94,253 both suppressed food intake in WT mice. 5-HT2C KO mice were insensitive to the hypophagic effects of mCPP but were more sensitive to CP-94,253-induced hypophagia than WT controls. mCPP induced a significant increase in post-prandial activity in 5-HT2C KO mice, but this effect was absent in 5-HT2C KO mice who were given CP-94,253. Data from WT mice, who were pre-treated with the 5-HT2C receptor antagonist SB 242,084 and then challenged with either mCPP or CP-94,253, were similar to those obtained from 5-HT2C KO mice. CONCLUSIONS: 5-HT2C-R and 5-HT1B-R activation are each sufficient to induce a hypophagic response. However, concurrent 5-HT2C-R inactivation can potentiate the hypophagic response to 5-HT1B-R activation, consistent with an inhibitory role for the 5-HT2C-R in behaviour mediated by the activation of other 5-HT receptors. These results also confirm that 5-HT1B-R activation alone cannot account for the hyperactive response of 5-HT2C KO mice to mCPP. PMID- 16470404 TI - Relevance of rodent models of intravenous MDMA self-administration to human MDMA consumption patterns. AB - RATIONALE: Despite decades of research specifying harmful effects produced by 3,4 methylenedioxymethamphetamine (MDMA; a principal component of 'ecstasy' pills), young people (and adults) continue to use it. In an attempt to model human MDMA consumption patterns, preclinical investigators have sought to establish reliable patterns of intravenous MDMA self-administration in rodents. OBJECTIVE: The objective of this report is to offer a critical review of published data (including our own novel findings) that reveal MDMA self-administration in rodents. RESULTS: The data indicate that MDMA serves as a reinforcer in rodents, though the responses are not similar to those previously reported for psychostimulants (i.e., cocaine). Important differences between rodent models and human use patterns include frequency of dosing and dosage exposure, routes of administration, tolerance that develops to MDMA after repeated exposure, polydrug use in humans but not by rodents, limits on the repertoire of behaviors that can be exhibited by rodents undergoing IV self-administration procedures, and the question of neurotoxicity as it relates to models of self-administration. CONCLUSIONS: While MDMA is not as potent a reinforcer as other drugs of abuse, the fact remains that young people and adults continue to use the drug, and therefore, additional research is needed to determine why drugs with low reinforcing effects continue to be abused. PMID- 16470406 TI - Glucose tolerance at age 58 and the decline of glucose tolerance in comparison with age 50 in people prenatally exposed to the Dutch famine. AB - AIMS/HYPOTHESIS: People who were small at birth have an increased risk of type 2 diabetes in later life. People who were in utero during the Dutch famine had decreased glucose tolerance and raised insulin concentrations at age 50. We aimed to evaluate whether prenatal famine exposure leads to more rapid progression of impaired glucose/insulin homeostasis with increasing age. METHODS: We performed an OGTT in 702 men and women at age 50 and in 699 men and women at age 58, all born as term singletons immediately before, during or after the 1944-1945 Dutch famine. RESULTS: People who had been exposed to famine in utero had significantly higher 120-min glucose concentrations at age 58 compared with people who had not been exposed to famine (difference=0.4 mmol/l, 95% CI 0.1 to 0.7, adjusted for sex and BMI). Glucose tolerance deteriorated between the age of 50 and 58. The unadjusted 120-min glucose concentrations rose by 0.2 mmol/l (95% CI 0.0 to 0.4), while 120-min insulin concentrations had increased by 64 pmol/l (95% CI 48 to 82). There were no differences in the rates of glucose and insulin level increase between the famine-exposed group and the unexposed group (p=0.28 for the difference in increase in glucose concentrations and p=0.09 for insulin concentrations). CONCLUSIONS/INTERPRETATION: Although we confirmed that undernutrition during gestation is linked to decreased glucose tolerance, the effect does not seem to become more pronounced at age 58 as compared with age 50. PMID- 16470407 TI - Trade-off in investment between dispersal and ingestion capability in phytophagous insects and its ecological implications. AB - In population ecology, dispersal plays a fundamental role, but is potentially costly. Traditionally, studies of phenotypic trade-offs involving dispersal focus on resource allocation differences between flight and reproduction. However, investments in dispersal may also result in reduced allocation to other "third party traits" (e.g. compensatory feeding) that are not directly associated with reproduction. Such traits remain largely uninvestigated for any phytophagous insect despite their importance for performance and survival. Using two wing dimorphic, phloem-feeding planthoppers, Prokelisia dolus and Prokelisia marginata that differ dramatically in dispersal abilities, we sought evidence for a trade off between investments in dispersal (flight apparatus) and ingestion capability (allocation to the esophageal musculature governing ingestion). Dispersal allows species to meet nutrient demands by moving to higher-quality resources. In contrast, enhanced investment in esophageal musculature increases ingestion capacity and allows phloem feeders to compensate for deteriorating plant nutrition on site. Our objectives were to compare differences in flight and feeding investment between P. dolus and P. marginata and between the wing forms of both species, and to compare ingestion capacity between the two species and wing forms. Morphometric and gravimetric measures of investment in flight versus feeding indicate that the sedentary P. dolus allocates more muscle mass to feeding whereas P. marginata invests more heavily in flight. Likewise, brachypters invest more in feeding and less in flight than macropters. The greater esophageal investment in P. dolus is associated with enhanced ingestion capacity compared to P. marginata. As a consequence, P. dolus is better equipped to meet on-site nutrient demands when faced with deteriorating plant quality than P. marginata, which must migrate elsewhere to do so. Notably, such third-party trade-offs place constraints on how insect herbivores cope with changing resources and set the stage for fundamental differences in population dynamics. PMID- 16470408 TI - Determining soil and ground water use of vegetation from heat pulse, water potential and stable isotope data. AB - A simple model of water uptake by vegetation is used to aid the discrimination of plant water sources determined with isotope data. In the model, water extracted from different soil depths depends on the leaf-soil potential difference, a root distribution function and a lumped hydraulic conductance parameter. Measurements of plant transpiration rate, and soil and leaf water potentials are used to estimate the value of the conductance parameter. Isotopic ratios in soil water and xylem are then used to constrain the root distribution. The model is applied to field measurements of transpiration, leaf water potential and 18O composition of xylem water on Corymbia clarksoniana, Lophostemon suaveolens, Eucalpytus platyphylla and Melaleuca viridiflora, and soil water potential and 18O composition of soil water to 8.5 m depth, in an open woodland community, Pioneer Valley, North Queensland. Estimates of the water uptake from various depths below the surface are determined for each species. At the time of sampling, the proportion of groundwater extracted by the trees ranged from 100% for C. clarksoniana to <15% for L. suaveolens and E. platyphylla. The advantages of the model over the traditional approach to determining sources of water used by plants using isotope methods are that it: (1) permits more quantitative assessments of the proportion of water sourced from different depths, (2) can deal with gradational soil water isotope profiles (rather than requiring distinct values for end-members), and (3) incorporates additional data on plant water potentials and is based on simple plant physiological processes. PMID- 16470409 TI - Pharmacokinetics of N-2-chloroethylaziridine, a volatile cytotoxic metabolite of cyclophosphamide, in the rat. AB - OBJECTIVES: The objectives of this study were to characterize pharmacokinetics of N-2-chloroethylaziridine (CEA) in the rat model and assess the in vivo fraction of total clearance of phosphoramide mustard (PM) that furnished CEA to circulation. METHODS: The disposition of CEA was investigated following separate intravenous (iv) administrations of PM, synthetic CEA, and their combination to the Sprague-Dawley rats. In addition, in rats receiving prodrug cyclophosphamide (CP), plasma concentrations of CP and its metabolites, 4-hydroxycyclophosphamide (HOCP), PM, and CEA, were simultaneously quantified using GC/MS and stable isotope dilution techniques. RESULTS: Following iv administration of synthetic CEA, concentrations of CEA declined biexponentially with the mean terminal half life and total body clearance of 47.5 min and 167 ml/min/kg, respectively. Urinary excretion of unchanged CEA was 0.164% of the administered dose. CEA was found to be the major circulating metabolite after iv administration of precursor PM to rats. The fraction of total clearance of PM that furnished CEA to circulation was estimated to be 100%, indicating virtually complete availability of the metabolite to circulation once formed. In rats administered with CP, PM exhibited the highest plasma and urinary concentrations compared to HOCP and CEA. CONCLUSIONS: For the first time, CEA was demonstrated to be an important in vivo metabolite of CP in the present study. In light of the poor permeability and in vivo stability of PM, the ultimate DNA alkylator, the findings obtained in this study suggested that CEA may contribute significantly to the overall antitumor activity of prodrug CP. PMID- 16470410 TI - Repression of telomerase reverse transcriptase mRNA and hTERT promoter by gambogic acid in human gastric carcinoma cells. AB - OBJECTIVES: To investigate the effects and potential mechanisms of gambogic acid (GA), a naturally occurring anticancer agent, on the expression and regulation of telomerase in human gastric carcinoma cells. METHODS: GA-induced inhibition of cell proliferation was evaluated by the commonly employed MTT assay on two human gastric carcinoma cell lines, MGC-803 and SGC-7901. Telomerase activity and hTERT mRNA expression were determined by telomeric repeat amplication protocol polymerase chain reaction and reverse transcription-polymerase chain reaction, respectively. The hTERT promoter activity was measured by luciferase assay. The expression of c-MYC, an apoptotic gene that modulates the expression of hTERT promoter, was quantified by Western blotting. RESULTS: The proliferation of human gastric carcinoma cell lines, MGC-803 and SGC-7901, was significantly inhibited with GA treatment. Both telomerase activity and hTERT mRNA expression were notably decreased in cells treated with GA. The activity of hTERT promoter and the expression of c-MYC were also remarkably decreased in GA-treated cells. CONCLUSION: This study demonstrated that GA treatment of human gastric carcinoma cell lines, MGC-803 and SGC-7901, significantly reduced the expression of c-MYC in a time- and concentration-dependent manner accompanied with the down regulation of the hTERT transcription and the ultimate reduction in telomerase activity. Our results indicate that the hTERT is a target of c-MYC activity and identify a feasible mechanism of GA's potent anticancer activity. PMID- 16470411 TI - Estimating radiation-induced cancer risks at very low doses: rationale for using a linear no-threshold approach. AB - The possible cancer risks caused by ionizing radiation doses of ~1 mSv or less are too small to be estimated directly from epidemiological data. The linear no threshold (LNT) approach to estimating such risks involves using epidemiological data at higher (but still low) doses to establish an "anchor point", and then extrapolating the excess cancer risk linearly down from this point to the low dose of interest. The study in this issue by Professor Tubiana and colleagues, summarizing a French Academy of Sciences report, argues that such LNT extrapolations systematically give substantial overestimates of the excess cancer risk at very low doses. We suggest that, to the contrary, even if there are significant deviations from linearity in the relevant dose range, potentially caused by the effects of inter-cellular interactions or immune surveillance, we know almost nothing quantitatively about these effects. Consequently, we do not know the magnitude, nor even the direction of any such deviations from linearity the risks could indeed be lower than those predicted by a linear extrapolation, but they could well be higher. PMID- 16470412 TI - A highly efficient in vitro plant regeneration system and Agrobacterium-mediated transformation in Plumbago zeylanica. AB - Plumbago zeylanica is a unique model for studying flowering plant gametogenesis, heterospermy, and preferential fertilization, yet understanding the control of related molecular mechanisms is impossible without efficient and reproducible regeneration and stable genetic transformation. We found three key factors for enhancing successful regeneration: (1) tissue source of explants, (2) combination and concentration of growth regulators, and (3) culture conditions. The highest frequency of shoot regeneration was achieved using hypocotyl segments cultured on MS basal medium supplemented with BA 2.0 mg/l, NAA 0.75 mg/l, adenine 50 mg/l and 10% (v/v) coconut milk under subdued light at 25+/-2 degrees C; under these conditions, each hypocotyl segment produced over 30 shoots, arising primarily through direct organogenesis after 3 weeks of culture. Regenerated shoots rooted easily on half-strength basal MS medium and were successfully established in the greenhouse. Using this tissue culture protocol, reporter gene GUS under the constitutive CaMV 35S promoter was introduced into P. zeylanica cells of petiole, cotyledon and hypocotyl with A. tumefaciens strains AGL1 and LBA4404. Transient expression was observed in all recipient tissues. Stable transgenic calli originating from petiole were obtained. PMID- 16470413 TI - The postural reduction in middle cerebral artery blood velocity is not explained by PaCO2. AB - In the normocapnic range, middle cerebral artery mean velocity (MCA Vmean) changes approximately 3.5% per mmHg carbon-dioxide tension in arterial blood (PaCO2) and a decrease in PaCO2 will reduce the cerebral blood flow by vasoconstriction (the CO2 reactivity of the brain). When standing up MCA Vmean and the end-tidal carbon-dioxide tension (PETCO2) decrease, suggesting that PaCO2 contributes to the reduction in MCA Vmean. In a fixed body position, PETCO2 tracks changes in the PaCO2 but when assuming the upright position, cardiac output (Q) decreases and its distribution over the lung changes, while ventilation (VE) increases suggesting that PETCO2 decreases more than PaCO2. This study evaluated whether the postural reduction in PaCO2 accounts for the postural decline in MCA Vmean). From the supine to the upright position, VE, Q, PETCO2, PaCO2, MCA Vmean, and the near-infrared spectrophotometry determined cerebral tissue oxygenation (CO2Hb) were followed in seven subjects. When standing up, MCA Vmean (from 65.3+/-3.8 to 54.6+/-3.3 cm s(-1) ; mean +/- SEM; P<0.05) and cO2Hb ( 7.2+/-2.2 micromol l(-1) ; P<0.05) decreased. At the same time, the VE/Q ratio increased 49+/-14% (P<0.05) with the postural reduction in PETCO2 overestimating the decline in PaCO2 (-4.8+/-0.9 mmHg vs. -3.0+/-1.1 mmHg; P<0.05). When assuming the upright position, the postural decrease in MCA Vmean seems to be explained by the reduction in PETCO2 but the small decrease in PaCO2 makes it unlikely that the postural decrease in MCA Vmean can be accounted for by the cerebral CO2 reactivity alone. PMID- 16470414 TI - Differential display analysis of gene expression in two immunologically distinct strains of Eimeria maxima. AB - Gene expression during sporulation and sporozoite excystation of two strains of Eimeria maxima was analyzed using the mRNA differential display technique. The two strains, the Guelph strain (GS) and a single sporocyst-derived strain (M6) from Florida, have been shown to be immunologically distinct. We isolated and cloned a 453-bp complimentary DNA (cDNA) fragment (GS-453) found only in GS sporozoites. In GS, this mRNA begins to be expressed during the earliest stages of oocyst sporulation and is continuously expressed up to and including in the excysted sporozoite. In all Northern blots, digoxigenin (DIG)-labeled GS-453 probe recognized an mRNA of approximately 1.6 kb from GS but not from RNA of M6. Southern blots using various endonucleases and probed with DIG-labeled GS-453 demonstrated that the genomes of both strains contained sufficiently similar sequences to permit hybridization with the probe, but the pattern of hybridization differed between the two strains. Extensive searches of the GenBank, European Molecular Biology Laboratory, and various apicomplexan expressed sequence tag databases using the DNA or inferred amino acid sequences of GS-453 cDNA clone did not identify similarity to any existing sequences. PMID- 16470415 TI - Multi-membrane-bound structures of Apicomplexa: II. the ovoid mitochondrial cytoplasmic (OMC) complex of Toxoplasma gondii tachyzoites. AB - Apicomplexa including the causative agents of toxoplasmosis and malaria reportedly possess one or few tubular-shaped mitochondria that permeate, more or less branched, throughout these unicellular parasites. Electron micrographs generated herein from serial-sectioned Toxoplasma gondii tachyzoites demonstrated, however, a greater diversity regarding both the shape of the cultured parasite's single mitochondrion and its sub-structural organization. Moreover, a unique subcellular construction was detected that basically comprised a pouch-shaped subdivision of the tachyzoite mitochondrion plus a fraction of parasitic cytoplasm enclosed therein. This composite assembling, termed ovoid mitochondrial cytoplasmic (OMC) complex, characteristically displayed a highly reduced matrix lumen of its mitochondrial border construction, which furthermore often failed to possess any cristae or contained tightly pleated cristae, thus creating a pouch-shaped multi-laminar wall of four or more membranous layers, respectively. Given this architecture, cross-sectioned OMC complexes of T. gondii tachyzoites frequently mimicked in size and shape the parasites' plastid-like organelle (apicoplast). Moreover, like the apicoplast, the OMC complex was often found adjacent to the tachyzoite's single Golgi complex and constantly located in close proximity to the outer membrane of the parasite's nuclear envelope. The T. gondii OMC complex differed, however, from the apicoplast in its exact fine structural organization and a stage-restricted presence that was apparently linked to mitochondrial growth and/or division. Any special function(s) possibly performed by the T. gondii OMC complex remains, nevertheless, to be elucidated. PMID- 16470416 TI - Physiological responses of insular wild black rat (Rattus rattus) to natural infection by the digenean trematode Fasciola hepatica. AB - Wild black rat Rattus rattus is regularly infected by the liver fluke Fasciola hepatica on Corsica. This report constitutes the only example of a murid rodent that plays an important epidemiological role for the Fasciolosis. We investigated the influence of such unusual parasite infection on black rat physiology by measuring its oxygen consumption at different ambient temperatures. Black rat energy requirements are influenced by body mass, temperature of the experiment and parasite infestation. The influence of the presence of F. hepatica was more pronounced for cold temperatures. The mean increase of 56% in oxygen requirements for infected rats is extremely high, indeed unexpected, according to previous knowledge. These high physiological constraints may be explained by the recent confrontation of the digenean and the rodent. PMID- 16470417 TI - Heterogeneity of class I and class II MHC sequences in Schistosoma mansoni. AB - We investigated the genetic variations in class I and class II major histocompatibility complex (MHC) genes of Schistosoma mansoni and the effects of host MHC genotypes. S. mansoni was maintained in combinations of two mouse strains with different MHC genotypes, and the MHC gene sequences of the cercariae were investigated. The detected class I MHC gene sequences were variable, with high similarity between the H-2D(b) murine host and the parasite. For other combinations, however, the parasite sequence was homologous to those of anthropoids. All class II MHC sequences detected in S. mansoni were homologous to those of anthropoids. Our results suggest that the genetic variation in the MHC sequences of S. mansoni is derived in part from the current host, indicating horizontal transfer of the sequences from mammal to parasite. PMID- 16470418 TI - Giant oesophageal fibrovascular polyp (2005:12b). AB - The authors report a case of a giant fibrovascular polyp of the oesophagus in which CT with multiplanar reformatting gave valuable information as to both location of the lesion as well as size and anatomical attachment proximally, which was pivotal in directing surgery. It was instructive in this case that initial endoscopy was reported to be normal. The lesion was well demonstrated by barium swallow. PMID- 16470420 TI - Exogenous nitric oxide inhibits IRS-1 expression in rat hepatocytes and skeletal myocytes. AB - Accumulative evidence has supported the role of nitric oxide (NO) in a variety of normal physiological functions as well as many pathological conditions. In this study, we examined the possible diabetogenicity of NO by measuring the expression of the insulin receptor substrate (IRS)-1 in rat hepatocytes and skeletal myocytes. IRS-1 is important in the insulin-mediated signal transduction pathway in both liver and skeletal muscle. Exogenous NO donated by S-nitroso-N acetylpenicillamine (SNAP) and S-nitrosoglutathione (GSNO) resulted in significant reduction in levels of IRS-1 in both cells, when compared to the insulin-stimulated control (p<0.001). Reversal to near normal levels was achieved using the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (carboxy-PTIO). SNAP was the more potent drug, and the skeletal myocytes were the more sensitive cells to the inhibitory effects of NO released from the drugs. These results provide further evidence that exogenous NO is a potent modulator of insulin-mediated signal transduction and may play a significant role in the pathogenesis of type 2 diabetes mellitus. PMID- 16470421 TI - BALLView: an object-oriented molecular visualization and modeling framework. AB - We present BALLView, an extensible tool for visualizing and modeling bio molecular structures. It provides a variety of different models for bio-molecular visualization, e.g. ball-and-stick models, molecular surfaces, or ribbon models. In contrast to most existing visualization tools, BALLView also offers rich functionality for molecular modeling and simulation, including molecular mechanics methods (AMBER and CHARMM force fields), continuum electrostatics methods employing a Finite-Difference Poisson Boltzmann solver, and secondary structure calculation. Results of these computations can be exported as publication quality images or as movies. Even unexperienced users have direct access to this functionality through an intuitive graphical user interface, which makes BALLView particularly useful for teaching. For more advanced users, BALLView is extensible in different ways. Owing to its framework design, extension on the level of C per thousand+ per thousand per thousand+ code is very convenient. In addition, an interface to the scripting language Python allows the interactive rapid prototyping of new methods. BALLView is portable and runs on all major platforms (Windows, MacOS X, Linux, most Unix flavors). It is available free of charge under the GNU Public License (GPL) from our website http://www.ballview.org. PMID- 16470422 TI - Postoperative Cox inhibitors and late prosthetic loosening--suspicion increases! PMID- 16470423 TI - Do non-steroidal anti-inflammatory drugs cause endoprosthetic loosening? A 10 year follow-up of a randomized trial on ibuprofen for prevention of heterotopic ossification after hip arthroplasty. AB - BACKGROUND: Non-steroidal anti-inflammatory drugs (NSAIDs) are known to be potent inhibitors of new bone formation. We investigated whether NSAIDs given at surgery influence the long-term results after total hip arthroplasty (THA). PATIENTS AND METHODS: We performed a 10-year follow-up on 142 of 144 patients who had taken part in a randomized trial on the preventive effects of the NSAID ibuprofen on heterotopic ossification after THA. 96 patients were treated with ibuprofen: 48 for 1 week postoperatively, 48 for 2 weeks postoperatively, and 48 patients were not treated. RESULTS: 13 patients had been revised. All revisions except 1 belonged to groups treated with ibuprofen. The 10-year risk for revision was significantly higher in the ibuprofen-treated patients (p = 0.05). Eleven of the revisions occurred due to fractures of the femur (2) or aseptic loosening (9), reasons that may be attributed to negative effects of ibuprofen. For these patients, the 10-year risk for revision was not statistically significantly different between treated and untreated patients (p = 0.08). In addition to the revised patients, 94 other patients were alive at the 10-year follow-up and 84 underwent radiographic examination. 9 loose prostheses were found radiographically, but these were equally distributed between ibuprofen-treated and untreated hips. INTERPRETATION: The high proportion of revisions in the ibuprofen groups, in combination with clinical and experimental evidence of inhibitory effects on new bone formation of NSAIDs, warrants further investigation of the effects of these drugs on prosthetic fixation. PMID- 16470424 TI - Drugs and fracture repair. PMID- 16470425 TI - Lipopolysaccharide impairs fracture healing: an experimental study in rats. AB - BACKGROUND: It has been shown that trauma causes translocation of lipopolysaccharide (LPS) endotoxins from the gut. LPS has been identified as a major bacterial bone resorbing factor. The effects of LPS on bone healing are therefore of clinical interest, as trauma involving fractures followed by sepsis is a clinical scenario. We investigated the effects of systemic and local administration of LPS on the healing of femoral fractures in rats. ANIMALS AND METHODS: In 3 groups, each consisting of 9 rats, a mid-diaphyseal osteotomy/fracture of the femoral bone was performed and then nailed. In one group of animals, LPS was applied intraperitoneally (systemically), and in another group, LPS was applied locally at the fracture site. The third group served as a control. The animals were killed after 6 weeks, and the mechanical characteristics of the healing osteotomies were evaluated. RESULTS: We found that LPS induced a hypertrophic and immature callus, as evaluated by bone mineral content and density. In the rats given LPS intraperitoneally, the mechanical strength characteristics were reduced, as evaluated by bending moment, rigidity, and energy absorption. INTERPRETATION: The rats given LPS intraperitoneally reflect a clinical situation with fracture trauma and endotoxinemia. Our findings indicate that endotoxinemia may impair the fracture healing processes. PMID- 16470426 TI - Removal or retention--will we ever know? The posterior cruciate ligament in total knee replacement. PMID- 16470427 TI - Retention versus removal of the posterior cruciate ligament in total knee replacement: a systematic literature review within the Cochrane framework. AB - BACKGROUND: There is no consensus as to whether to use a posterior cruciate ligament (PCL) retaining design or a posterior-stabilized design for total knee arthroplasty. The objective of this study was to establish the difference in functional, clinical, and radiological outcome between retention and removal of the PCL. METHODS: We conducted a search in Medline, EMBASE, the Cochrane database, and Current Contents, along with reference checks and citation tracking. Randomized controlled trials were selected and methodological quality was assessed with the van Tulder and Jadad checklists by 2 independent reviewers. RESULTS: We found 8 randomized controlled trials. 2 treatment options were compared against PCL retention: PCL removal without post and cam mechanism (2 studies), and posterior-stabilized design (5 studies). 1 study included all 3 options. Range of motion was found to be 8 degrees higher (105 degrees vs. 113 degrees ) in the posterior-stabilized group compared to the PCL retention group (p = 0.01, 95% CI (1.7, 15)). INTERPRETATION: These results should be interpreted with caution, as the methodological quality of the studies was highly variable. Suggestions are given to improve future research on this specific aspect of knee arthroplasty. PMID- 16470428 TI - All-polyethylene vs. metal-backed tibial component in total knee arthroplasty-a randomized RSA study comparing early fixation of horizontally and completely cemented tibial components: part 1. Horizontally cemented components: AP better fixated than MB. AB - BACKGROUND: Controversy still exists as to whether to mount the tibial bearing on a metal baseplate or not. Furthermore, the tibial component can be either horizontally or completely cemented. We evaluated metal backing versus all polyethylene using horizontal cementing technique. PATIENTS AND METHODS: We randomized 40 patients with knee arthrosis (40 knees) to 2 groups: cemented total knee arthroplasty (AGC, Biomet) using either all-polyethylene (AP) or metal backed (MB) tibial components (n = 20 for each group). All tibial implants had a total thickness of 8 mm. We used proximal cement-ing, including only the horizontal part of the tibia with avoidance of the stem-bone interface. RESULTS: The positioning of the implants, as judged by the HKA angle, showed an average neutral alignment in both groups. Preoperatively and after 2 years, there was no statistically significant difference between the HSS scores in both groups (p = 0.6 and 0.4). After 2 years, the metal-backed components had rotated more around the longitudinal axis, median 0.5 degrees vs. 0.2 degrees (internal or external rotation, p = 0.002), and showed increased maximum total point motion, median 1.0 vs. 0.4 mm (maximum migration or MTPM, p = 0.003). INTERPRETATION: Our study indicates that better fixation can be obtained with the all-polyethylene design if proximally cemented thin components are used. PMID- 16470429 TI - All-polyethylene vs. metal-backed tibial component in total knee arthroplasty-a randomized RSA study comparing early fixation of horizontally and completely cemented tibial components: part 2. Completely cemented components: MB not superior to AP components. AB - BACKGROUND: Uncertainty exists as to whether metal backing (MB) of the tibial component is better than an all-polyethylene component (AP). This is valid for both horizontally and completely cemented components. We evaluated completely cemented MB vs. AP (Part 2). PATIENTS AND METHODS: In a randomized study, 39 patients (40 knees) with knee arthrosis were operated with cemented low conforming total knee arthroplasty (AGC, Biomet) with a tibial component of uniform thickness (8 mm), cemented both beneath the tibial tray and around the stem. 20 patients had an all-polyethylene (AP) tibial component and 20 patients had an identical but metal-backed (MB) tibial component. We used clinical examination and radiostereometric analysis (RSA) to evaluate the hypothesis that MB improves component fixation. Fixation was evaluated using RSA up to 2 years after surgery. Clinical assessment was performed preoperatively and after 2 years using the Hospital for Special Surgeons (HSS) score. RESULTS: We found no differences in micromotion, and no differences in clinical scores could be detected between the groups at any time point. INTERPRETATION: Our findings indicate that there was equal initial fixation of the AP and MB stemmed monobloc components when they were cemented beneath the tibial plateau and around the stem. PMID- 16470430 TI - Revision in previously satisfied knee arthroplasty patients is the result of their call on the physician, not on pre-planned follow-up: a retrospective study of 181 patients who underwent revision within 2 years. AB - BACKGROUND: Degree of satisfaction with a knee arthroplasty is said to be correlated to reduced pain and better function. During a validation of the Swedish Knee Arthroplasty Register in 1997, previously operated patients were asked how satisfied they were with their knee. A subgroup of "satisfied" patients was identified who underwent revision within 2 years of having expressed satisfaction. Our aim was to study the revision diagnosis, to determine whether the problem leading to revision had been discovered as a result of routine follow up, and also to find out when the symptoms leading to revision had started. METHODS: We retrospectively studied the medical records of 181 patients (181 knees), with a median age of 74 (31-88) years. 68% were women and the median time between primary operation and revision was 8 (3-21) years. RESULTS: Aseptic loosening (74/181) was the most common diagnosis. 2 cases were revised as a result of routine follow-up. 44% of the medical records included reports of pain in the replaced knee prior to answering the satisfaction questionnaire. INTERPRETATION: Few patients were admitted to knee revision surgery due to medical findings discovered during routine follow-up. The term "satisfaction" must be interpreted with care, as it seems to have a more complex meaning for the patients than absence of knee pain. PMID- 16470431 TI - Measurement of bone mineral density is possible with standard radiographs: a study involving total knee replacement. AB - BACKGROUND: Measured bone density correlates well with the mechanical properties of the bone. Our objective was to evaluate optical densitometry as a method of measuring the periprosthetic bone density of the tibial platform in total knee replacement using serial dual-energy X-ray absorptiometry (DXA) as the reference test. METHODS: 30 patients who underwent a cemented total knee replacement were followed up for over 2 years. Standard radiographs of the knee were obtained at 12 and 24 months. These were photographed with a digital camera and enhanced with image processing software. A DXA scan was performed at the same time as the reference method. 3 regions of interest were defined under the tibial plateau (medial, lateral and stem positions). RESULTS: After 24 months of follow-up there was a significant decrease in density in the 3 regions, both with the optical and DXA methods. The concordance between methods was studied using Bland and Altman plots, Cronbach's alpha and intraclass correlation coefficients resulted in values from 0.72 to 0.87, depending on the region and the follow-up time. INTERPRETATION: Quantification of optical density values with a standardized measurement system on conventional radiographs is a reliable and efficient method of determining the bone mineral density. PMID- 16470432 TI - Comparison of distribution, agreement and correlation between the original and modified Merle d'Aubigne-Postel Score and the Harris Hip Score after acetabular fracture treatment: moderate agreement, high ceiling effect and excellent correlation in 450 patients. AB - BACKGROUND: In acetabular fracture treatment, 3 disease-specific outcome scores are mainly used: the original and modified Merle d'Aubigne-Postel Score, and the Harris Hip Score. METHODS: The original and modified Merle d'Aubigne-Postel Score and the Harris Hip Score were recorded in 1,153 follow-ups of 450 patients. 492 follow-ups were excluded because factors other than the acetabular fracture were found to affect the outcome scores. This gave 661 patient records for the study. RESULTS: The Spearman correlations were between 0.81 and 0.89. The quartile analyses showed Kappa agreement between 0.45 and 0.55. About 40% of the observations were classified into another quartile when switching from one outcome score to another. The 25th and 50th percentiles comprised 85% and 95% of the total numeric scores, respectively, while the 75th percentiles showed ceiling value (100% of the maximum) in all 3 scores. INTERPRETATION: Despite the excellent overall correlation between the outcome scores, the Kappa agreements were only moderate. The scores were all skewed in distribution with considerable ceiling effects that could limit their clinical use. The scores did not capture any differences in 25% of the observations at the upper end of the scales. PMID- 16470433 TI - Transitional transverse acetabular fractures: differences between fractures with a large posterio-superior fragment and the inverse T-fracture--a report of 10 unusual cases. AB - BACKGROUND: Classification of fractures is crucial in decision making and planning of acetabular surgery. Transverse fractures with secondary vertical fracture lines--which constitute either a large posterio-superior fragment (floating dome) or an inverse T with the posterior ileum attached to the axial skeleton--have not been described in detail in the literature. METHODS: All acetabular fractures at Ulleval University Hospital have been recorded prospectively since 1993 and classified according to Judet/Letournel. RESULTS: In 10/449 fractures (2%) there was a transverse fracture line through the acetabulum, with an additional vertical fracture line ascending either to the iliac crest or the SI-joint. In 6 of these fractures the vertical line started within the acetabulum, and the posterior part of the ileum with part of the articular surface was attached to the axial skeleton. In 4 fractures the vertical fracture line started posterior to the joint, constituting a very large posterio superior fragment without any articular surface (a floating dome). INTERPRETATION: Open reduction and internal fixation of inverse T-fractures and transverse fractures with a floating dome require different surgical approaches. The latter can be treated through a single approach alone, while the inverse T fracture may require extensile or combined approaches for adequate reduction and fixation. PMID- 16470434 TI - Staged bilateral total hip arthroplasty using rough and smooth surface femoral stems with similar design: 10-year survivorship of 48 cases. AB - BACKGROUND: Various studies have reported good long-term results using femoral stems with either smooth or rough surfaces. In this retrospective cross-sectional survivorship study, we reviewed the 10-year results of 51 bilateral staged cemented total hip arthroplasties using the Harvard or the Charnley femoral stems which have almost similar geometry but a different surface finish. METHODS: 51 patients were reviewed at median interval of 10 (Harvard group) and 11 years (Charnley group) after the primary operation. We evaluated cement mantle thickness, alignment of the components, presence of radiolucent lines, and aseptic loosening. Kaplan-Meier analysis was performed to calculate the survival rate using various endpoints. RESULTS: 8 hips in the Harvard group were revised for aseptic loosening of the femoral component at a median interval of 6.3 years after the primary procedure. 3 hips in the Charnley group were revised for aseptic loosening of the femoral and acetabular components between 10 and 11 years after the primary procedure. The 10-year survival rate for the femoral component using revision surgery for aseptic loosening as an endpoint was 80% (95% CI: 31-42) and 95% (95% CI: 44-47) in the Harvard and the Charnley group, respectively. Cox regression analysis did not reveal any statistically significant effect of various radiographical parameters on the survival rate (p < 0.05). INTERPRETATION: Our results demonstrate that in the group of patients studied, the femoral stem component with the matt surface finish had less satisfactory 10-year survival than the femoral stem of similar design which had a smooth surface finish. PMID- 16470435 TI - Regional variation in incidence of primary total hip arthroplasties and revisions in Denmark, 1996-2002. AB - BACKGROUND: We examined incidence rates (IR) of total hip arthroplasty (THA) in Denmark according to county and type of hospital, and investigated possible factors associated with any variations. METHODS: Through the Danish Hip Arthroplasty Registry, we identified all cases of primary THAs (n = 37,144) and revisions (n = 6,446) between 1996 and 2002. Unadjusted and age- and gender standardized, hospital type- and county-specific IRs were calculated. Using multiple regression analysis, we examined possible factors associated with the variations. RESULTS: Over the study period, age- and sex-standardized IRs of primary THAs for university and other hospitals increased by 10% and 40%, respectively. IRs of revisions increased for other hospitals by 33%, but decreased for university hospitals by 30%. Unadjusted IRs varied among counties (n = 15), which remained despite age- and gender-standardization. The IR ratio was 1.4 (95% CI: 1.3-1.6) when comparing the counties with the highest and lowest IRs. No association was found between IRs of THA and the number of orthopedic surgeons per 100,000 inhabitants, proportion of patients with primary arthrosis, preoperative Harris Hip Score, hospital costs, the counties'Gross Domestic Product, or population density. INTERPRETATION: Substantial regional differences in the IRs of THA procedures were found. We found no associations between IRs and a range of patient- and healthcare system-related factors. PMID- 16470436 TI - Validation of data in the Norwegian Arthroplasty Register and the Norwegian Patient Register: 5,134 primary total hip arthroplasties and revisions operated at a single hospital between 1987 and 2003. AB - BACKGROUND: The usefulness of a national medical register relies on the completeness and quality of the data reported. The data recorded must therefore be validated to prevent systematic errors, which can cause bias in reports and study conclusions. PATIENTS AND METHODS: We compared the number of hip replacements reported to the Norwegian Arthroplasty Register (NAR), 1987-2003, and to the Norwegian Patient Register (NPR), 1999-2002, with data recorded at a local hospital. The date of operation and the index hip were further validated to find inaccurately recorded data in the NAR. Kaplan-Meier estimated survival curves were compared to evaluate the possible influence of missing data. RESULTS: Of 5,134 operations performed at a local hospital, 19 (0.4%) had not been reported to the NAR. Completeness of registration was poorer for revisions (1.2%) than for primary operations (0.2%). Among 86 Girdlestone revisions (removal of the prosthesis only), 9 (11%) had not been reported to the NAR. Missing data on revisions, however, had only a minor influence on survival analyses. The date of the operation had been recorded incorrectly in 56 cases (1.1%), and the index hip in 12 cases (0.2%). The surgeon was responsible for 85% of these errors. Comparisons with data reported to the NPR, 1999-2002, showed that 3.4% of operations at the local hospital had not been reported to the NPR. INTERPRETATION: Only 0.4% of the data from a local hospital was missing in the NAR, as opposed to the NPR where 3.4% was missing. The information recorded in the NAR appears to have been valid and reliable throughout the entire period, and provides an excellent basis for clinically relevant information regarding total hip arthroplasty. PMID- 16470437 TI - Tranexamic acid reduces blood loss in cemented hip arthroplasty: a randomized, double-blind study of 39 patients with osteoarthritis. AB - BACKGROUND: Tranexamic acid has been found to reduce blood loss and the need for blood transfusions in knee arthroplasty. In hip arthroplasty, the benefit of tranexamic acid is not as clear. PATIENTS AND METHODS: In a randomized, double blind study, 39 patients with primary cemented hip arthroplasty for osteoarthritis were divided into two groups; one receiving tranexamic acid and the other not receiving it. Tranexamic acid was given in a dose of 10 mg/kg before the operation and twice thereafter, at 8-hour intervals. RESULTS AND INTERPRETATION: Total blood loss was smaller in the tranexamic acid group than in the control group. No thromboembolic complications were noticed. Tranexamic acid appears to be an effective and economic drug for reduction of blood loss in cemented primary hip arthroplasty for osteoarthritis. PMID- 16470438 TI - The Bernese periacetabular osteotomy: clinical, radiographic and mechanical 7-15 year follow-up of 26 hips. AB - BACKGROUND: The Bernese periacetabular osteotomy is used in dysplastic hips to increase the load-bearing area of the hip and to prevent osteoarthritis. The aim of our work was to determine the contact hip stress before and after the osteotomy and to compare the relief of stress with the long-term radiographic and clinical outcome. PATIENTS AND METHODS: We followed 26 dysplastic hips (26 patients) for 7-15 years after the index operation. Clinical evaluation was based on the WOMAC score, osteoarthrosis was evaluated with the Tonnis classification, the angles of lateral (CE) and anterior (VCA) femoral coverage were measured, and biomechanical parameters were studied. RESULTS: Periacetabular osteotomy increased the mean CE from 15 degrees to 37 degrees , and the mean VCA from 22 degrees to 38 degrees . The mean normalized peak contact stress was reduced from 5.2 to 3.0 kPa/N. Four hips required total hip arthroplasty after an average of 4.5 years, 8 hips showed considerable arthrosis progression, and 14 hips had no or mild arthrosis at follow-up. Preoperative WOMAC score, preoperative Tonnis grade and postoperative normalized peak contact stress were the most important predictors of outcome. INTERPRETATION: The Bernese periacetabular osteotomy improves the mechanical status of the hip. Long-term success depends on the grade of arthrosis preoperatively and on the magnitude of operative correction of the contact hip stress. PMID- 16470439 TI - Children with acute Perthes' disease have asymmetrical lower leg growth and abnormal collagen turnover. AB - BACKGROUND: Abnormalities in distal growth and low levels of insulin-like growth factor (IGF)-I have been reported in children with Perthes' disease. Our aim was to establish whether the acute phase of Perthes' disease is associated with abnormalities of growth, of bone or of collagen turnover. METHODS: We performed a cross-sectional study of 15 children (3-11 years of age, 13 boys) at acute presentation and a longitudinal cohort study of 9 children. We measured (1) the lengths of both lower legs (by knemometry) at weeks 1, 2, 6 and 12, (2) height and weight at presentation and at the second-year follow-up, and (3) levels of IGF-I, IGFBP-3, collagen markers and bone alkaline phosphatase at weeks 1 and 12, and in year 2. RESULTS: Height SD scores were normal at presentation but declined thereafter. Lower leg growth was not impaired at presentation but was asymmetrical, ceased during weeks 2-6, and then resumed symmetrically. Patients had persistently low IGF-I, low soft tissue collagen synthesis and enhanced collagen breakdown compared with age- and sex-related reference data. Markers of bone formation increased during follow-up. INTERPRETATION: Acute changes in lower leg growth reflected differential weight bearing, then immobilization and remobilization. Persistently low IGF-I may have contributed to low soft tissue collagen synthesis and growth. Changes in bone formation markers most likely reflected bone healing. PMID- 16470440 TI - The frontal pelvic plane provides a valid reference system for implantation of the acetabular cup: spatial orientation of the pelvis in different positions. AB - BACKGROUND: The frontal pelvic plane has traditionally served as the reference plane for implantation of the acetabular cup during total hip arthroplasty, with referencing performed with the patient supine on the operating table. During daily activities in an upright position, the frontal pelvic plane changes from a horizontal to a vertical orientation. If this change in orientation is accompanied by a substantial change in pelvic inclination angle, it would mean that the use of the frontal pelvic plane as a reference plane for implantation of the acetabular cup would not be valid for proper alignment of the cup. To evaluate this possibility, we measured the change of inclination of the pelvis from the supine to the standing position. SUBJECTS AND METHODS: We evaluated 120 patients, first positioned in a standing position and then supine on a table. Three pelvic landmarks were digitized percutaneously, and the spatial coordinates were calculated with regard to pelvic orientation in the horizontal and the vertical plane. RESULTS: We found a mean inclination of 6.7 degrees in the standing position and 5.6 degrees in the supine position. Patients who were more than 60 years of age who did not have coxarthrosis had a greater inclination angle (8.7 degrees ) while standing. Pelvic orientation was stable with regard to the supine and standing positions. These results were independent of sex, level of arthrosis, or status after implantation of a total hip replacement. INTERPRETATION: The frontal pelvic plane is a valid reference plane for implantation of the acetabular cup. PMID- 16470441 TI - Intraarticular lesions in traumatic anterior shoulder instability: a study based on the results of diagnostic imaging. AB - BACKGROUND: Diagnostics of intraarticular lesions is important in the treatment of traumatic anterior shoulder instability. PATIENTS AND METHODS: We studied 30 patients: 26 men and 4 women. For all patients preoperative examinations involved arthrography, CT arthrography, MR arthrography, and 3D-CT. RESULTS: The articular capsule adhered to the margin of the glenoid in only 3 cases. In the remaining 27 patients, the joint capsule was detached, showing medial translation. Hill-Sachs lesions were present in 24 cases. Bankart lesions were detected in all cases, and they extended over 2-5 h. Osseous Bankart lesions were detected in 13 cases. MR arthrography revealed the articular labrum in all cases, but AIGHL in only 16 cases. Bankart lesions were found in all cases. When the patients were divided according to the frequency of dislocation, there was a correlation between both distance of the detached joint capsule and visualization of AIGHL and the frequency of dislocation. INTERPRETATION: From this study, the frequency of dislocation was higher in the patients with severe detachment of the joint capsule, in whom AIGHL could not be visualized by MR arthrography. PMID- 16470442 TI - Nonoperative treatment of displaced supracondylar fractures in children: Rigault type 2 fractures. AB - BACKGROUND: Current opinion in the medical literature concerning displaced supracondylar fractures of the distal humerus recommends pinning because with flexion braces there is a risk of both secondary displacement and Volkmann syndrome. PATIENTS AND METHODS: We analyzed 84 children with displaced supracondylar fractures. According to Rigault's classification, 30 children had grade 2 fractures, 21 had grade 3, 28 had grade 4 and 5 had multiple fragments, which were thus outside this classification. Fractures that could be reduced to a stable position under general anesthesia were treated with a posterior long arm splint with an average elbow flexion of 113 degrees (90-140). This technique was applied in 28 of the grade 2 fractures and in 4 of the grade 3 fractures, but in none of the grade 4 fractures. RESULTS: Of the 4 cases of Rigault grade 3 fractures treated nonoperatively, 3 had to be re-reduced and 1 needed an operation later on for varus correction. Of the 28 Rigault grade 2 fractures, 27 showed excellent results, and 1 had a good result. We advise nonoperative treatment in type 2 supracondylar fractures if stable reduction is achieved. PMID- 16470443 TI - Minimally invasive surgical techniques for irreducible supracondylar fractures of the humerus in children. AB - BACKGROUND: Treatment of severely displaced supracondylar fracture of the humerus in children remains a challenge. We retrospectively assessed the outcomes of the minimally invasive open reduction techniques used in our institutions for the treatment of grade 3 supracondylar fractures in children which could not be reduced by closed manipulation. PATIENTS AND METHODS: 78 children (58 boys) with severely displaced supracondylar fractures of the humerus and severe swelling were treated with either a minimal incision in the anticubital fossa and manipulation of the distal fragment with the thumb, or a small stab incision and manipulation of the fracture fragment with a small-sized suction tip. The fractures were stabilized with 1.6-mm Kirschner wires. RESULTS AND INTERPRETATION: The outcome after a follow-up of at least 3 years was excellent in 76 cases and poor in 2 cases. No scar contractures or other complications were observed. These techniques seem safe and effective in the treatment of irreducible grade 3 supracondylar fractures of the humerus in children. PMID- 16470444 TI - Treatment of sequelae of radial head fractures with a bipolar radial head prosthesis: good outcome after 1-4 years follow-up in 11 patients. AB - BACKGROUND: In the presence of additional disruption of the distal radioulnar ligaments, the interosseous membrane, or the lateral- and/or medial collateral ligament, radial head fractures treated by resection will result in valgus elbow instability, proximal radial migration and/or posterolateral rotatory instability. Radial head replacement has been used to treat or prevent this. We report our experience with the Judet CRF II radial head prosthesis. PATIENTS AND METHODS: We treated 11 patients with a bipolar radial head prosthesis because of elbow instability after previous treatment for Mason-Johnston type III or IV radial head fractures. The outcome was assessed clinically using two standardized elbow function assessment scales, and radiographically after a mean follow-up of 2 years. RESULTS: Clinical outcome was either good or excellent in all patients; all elbows were stable. Radiographically, there were no signs of loosening, fracture or heterotopic ossification. 2 patients required reoperation for subluxation of the prosthesis; both were treated by reducing the size of the modular head of the prosthesis. There was erosion of the capitellum in 1 patient. INTERPRETATION: Bipolar radial head replacement can be used successfully for treatment of the sequelae of radial head fractures. The long-term outcome is, however, unknown. PMID- 16470445 TI - Marked initial displacement predicts complications in physeal fractures of the distal radius: an analysis of fracture characteristics, primary treatment and complications in 109 patients. AB - BACKGROUND: Factors predicting redisplacement in the cast and early complications in distal radial physeal injuries have not been analyzed before. PATIENTS AND METHODS: We analyzed 109 consecutive children with displaced physeal fractures of the distal radius treated by manipulation under anesthesia RESULTS: Acute carpal tunnel syndrome developed in 2 patients. Posttraumatic swelling necessitating trimming, splitting or removal of the primary circular cast occurred in one-sixth of the patients. Half of the fractures healed in malunion despite an anatomic primary reduction in 85% of the cases. According to logistic regression models, marked initial malposition of the fracture (< 50% displacement or < 20% angulation) was an independent risk factor for complications and redisplacement. Non-anatomic reduction of the fracture was an additional independent risk factor for redisplacement. INTERPRETATION: The risk of an acute carpal tunnel syndrome should be remembered in patients with marked primary displacement of the fracture. To avoid redislocation, pin fixation of the fracture in patients close to skeletal maturity should be considered if there is a marked initial malposition of the fracture, or if fully anatomic reduction cannot be achieved. PMID- 16470446 TI - Two-stage reconstruction with free vascularized soft tissue transfer and conventional bone graft for infected nonunions of the tibia: 6 patients followed for 1.5 to 5 years. AB - BACKGROUND: Vascularized soft tissue transfer may give better results of treatment of infected nonunions of the tibia. METHODS: 6 patients with infected nonunion of the tibia and combined soft tissue (70-170 cm(2)) and bony (5-8 cm) defects underwent staged reconstruction. Initial surgery consisted of soft tissue and bone debridement, external fixation, filling of the bony defect with a gentamicin-impregnated cement spacer, and reconstruction of the soft tissue with a free microsurgical muscle flap and skin graft. Second-stage surgery consisted of removal of the cement spacer and osseous reconstruction with nonvascularized bone graft. RESULTS: All patients except 1 achieved full weight-bearing and radiographic consolidation after 7-10 months. This patient required repeated bone grafting and internal plate fixation to heal. There were no cases of recurrence of infection at the latest follow-up, after a mean of 3 (1.5-5) years. INTERPRETATION: Staged reconstruction with free vascularized soft tissue transfer and conventional bone grafting within a cement-induced membrane is a low-risk surgical strategy resulting in a high rate of bone healing. PMID- 16470447 TI - Ankle arthrodesis by compression screws in rheumatoid arthritis: primary nonunion in 9/35 patients. AB - INTRODUCTION: Today, percutaneous or open arthrodesis of the ankle, using one or several screws for fixation, is a common method for treatment of the rheumatic ankle. However, there is very little information in the literature on the reliability of the method. METHODS: We performed a retrospective radiographic and clinical study on 35 ankles of 35 patients. Function was evaluated using the Mazur and AOFAS scores. For evaluation of activity of the rheumatic disease, we used the HAQ score. RESULTS: 31 ankles had healed--26 at the first attempt and 5 after repeat arthrodesis. There was no difference between 13 cases operated on percutaneously and 22 cases operated on with open technique with respect to radiographic healing. Mean AOFAS total score was 56 of possible 86 points. The AOFAS total score correlated with the severity of the rheumatic disease. 20 patients were satisfied with the result, 12 were somewhat satisfied and 3 patients were dissatisfied although the ankle in 2 of these 3 patients had fused. INTERPRETATION: The use of compression screws for fusion of the rheumatic ankle does not appear to give acceptable results regarding healing and function. PMID- 16470448 TI - Long-term results of the Weber operation for chronic ankle instability: 37 patients followed for 20-30 years. AB - BACKGROUND: The Weber operation is an anatomical reconstruction of the anterior talofibular ligament with the plantaris tendon. Few long-term studies have been published. METHODS: We evaluated 40 ankles in 37 patients (19 women) at an average of 24 years after the procedure. RESULTS: At follow-up, symptoms were present to vary-ing degrees in half of the ankles, but 32 patients were satisfied with the result and approximately two-thirds of the patients had a good or excellent result with the Karlsson and Good scores. Only 3 ankles had secondary arthrotic changes with narrowing (2 patients) or disappearance (1 patient) of the joint space. INTERPRETATION: We conclude that the Weber procedure is a good alternative for treatment of chronic anterolateral ankle instability when a direct anatomical reconstruction is not feasible, or has failed. PMID- 16470449 TI - Long-term follow-up of 15 patients with non-metastatic Ewing's sarcoma and a skip lesion. AB - BACKGROUND Skip lesions in Ewing's sarcoma of the bone seem to be rare; to our knowledge only 7 cases have been published in the English medical literature. METHODS: We retrospectively reviewed imaging and histological data relating to 235 patients with non-metastatic Ewing's sarcoma of the bone who participated in the cooperative Ewing's sarcoma study (CESS 86 and CESS 91), and we identified 15 patients with a skip lesion at diagnosis. RESULTS: The skip lesion was located in the same bone as the primary tumor in 13 patients, and in an adjacent juxtaarticular bone in 2 cases. The average follow-up was 11 years. Despite aggressive treatment including surgery in all cases, tumor relapse occurred in 9 patients, and 7 of these patients died due to metastatic disease. INTERPRETATION: Skip lesions in patients with otherwise non-metastatic skeletal Ewing's sarcoma may be of the same consequence as the molecular detection of marrow metastases and possibly confer a worse prognosis. Newer imaging modalities (for example PET) and careful staging work-up may indicate that skip metastases in Ewing's sarcoma are more common than previously suspected. PMID- 16470450 TI - Polymethylmethacrylate causes prolonged pulmonary hypertension during fat embolism: a study in sheep. AB - BACKGROUND: Fat embolism (FE), the release of bone marrow contents into the circulation and the subsequent cardiovascular changes, is still a potentially fatal com-plication during orthopedic surgery. Different causative factors have been suggested, but the exact pathomechanism of FE still remains unclear. We investigated the role of polymethylmethacrylate (PMMA) in FE during vertebroplasty in sheep. METHODS: In 8 sheep, two vertebral bodies were augmented alternatively with PMMA or bone wax. Pulmonary and cardiovascular parameters were monitored during the procedure. RESULTS: The peak response was similar for both groups and characterized by hypotension, a drop in cardiac output and pulmonary hypertension. However, the recovery in pulmonary arterial pressure and pulmonary vascular resistance was quicker in the wax group. INTERPRETATION: The injection of PMMA may cause prolonged pulmonary hypertension during vertebro-plasty and also arthroplasty. Surgeons should be aware of this potential cardiovascular complication, especially in patients with impaired pulmonary and cardiovascular function. PMID- 16470451 TI - No adverse effects of bone compaction on implant fixation after resorption of compacted bone in dogs. AB - BACKGROUND: A new bone preparation technique, compaction, has been shown to enhance initial implant fixation. However, short-term compaction has resulted in more non-vital bone being in contact with the implant. Also, compaction may result in inferior long-term implant fixation as the compacted non-vital bone at the bone-implant interface is resorbed. METHODS: We tested the hypothesis that compaction would result in inferior implant fixation after 10 weeks of weight bearing. We compared compaction with the conventional bone removal technique (drilling) for (1) porous coated titanium (Ti) implants inserted exact-fit into medial femoral condyles, and for (2) hydroxy-apatite (HA) porous coated implants inserted press-fit into lateral femoral condyles. In each of 8 dogs, we prepared the implant cavities of one knee joint with drilling, and the other with compaction. Implants were tested mechanically to failure by push-out test, and histomorphometry was done. RESULTS: For all specimens, non-vital bone implant contact contributed very little to the total bone implant contact. Inferior mechanical or histological implant fixation with compaction was not found for either Ti implants or HA implants. INTERPRETATION: Compaction does not appear to result in inferior implant fixation as the compacted bone at the bone implant interface is resorbed. PMID- 16470452 TI - The cellular origin of cartilage-like tissue after periosteal transplantation of full-thickness articular cartilage defects: an experimental study using transgenic rats expressing green fluorescent protein. AB - BACKGROUND: Periosteal transplantation is commonly used for the treatment of articular cartilage defects. However, the cellular origin of the regenerated tissue after periosteal transplantation has not been well defined. The objective of this study was to investigate the cellular origin of the regenerated tissue after periosteal transplantation. METHOD: Free periosteum was harvested from the tibia of 10-week-old adolescent enhanced green fluorescent protein (GFP-) expressing transgenic Sprague Dawley (SD) rats and was transplanted to full thickness articular cartilage defects of the patellar groove in normal 10-week old adolescent SD rats. The periosteum was sutured to the defect with the cambium layer facing the joint cavity. 8 SD rats were killed at 4 weeks and 8 SD rats were killed at 8 weeks after surgery. The repaired tissue was assessed histologically and histochemically. GFP-positive cells derived from the donor periosteum could easily be detected in the repaired tissue by use of a fluorescent microscope. RESULTS: At both 4 and 8 weeks after transplantation, the entire area of the defects had been repaired, with the regenerated tissue being well stained histologically with safranin-O. Most cells in the whole area of the regenerated tissue were GFP-positive, indicating that very few of the cells were GFP-negative cells originating from the recipient rats. INTERPRETATION: This experiment demonstrates that most cells in regenerated tissue after periosteal transplantation using adolescent animals do not originate from recipient cells but from the periosteal cells of the donor. PMID- 16470454 TI - Pseudo-reduction of an acute anterior dislocation of the shoulder--a case report. PMID- 16470453 TI - Glucocorticoids suppress proteoglycan production by human tenocytes. AB - BACKGROUND: The role of glucocortiocid injection therapy in spontaneous tendon rupture is controversial. We hypothesized that glucocorticoids suppress proteoglycan production in tendon and studied the in vitro effects of dexamethasone and triamcinolone on proteoglycan production by cultured human tenocytes. MATERIAL AND METHODS: We obtained primary cultures of human tenocytes from explants of healthy human patellar tendon. The human tenocytes were treated with 1 microM dexamethasone or 1 microM triamcinolone. The amount of proteoglycan production was measured by 35S-sulfate incorporation assay and compared with control cultures. The reversibility of the effect of dexamethasone by co incubation with 10 ng platelet-derived growth factor (PDGFBB) was also tested. RESULTS: Treatment with 1 microM triamcinolone reduced the amount of 35S-sulfate incorporation to 80% of control cultures (p = 0.007), whereas 1 microM dexamethasone reduced it to 72% (p = 0.01). Co-incubation of 10 ng/mL PDGFBB with 1 microM dexamethasone returned the 35S-sulfate incorporation to a level that was significantly higher than for dexamethasone treatment alone (108%; p = 0.01). INTERPRETATION: Glucocorticoids suppressed proteoglycan production in cultured human tenocytes. The suppression by dexamethasone was reversed by simultaneous addition of PDGFBB. Suppressed proteoglycan production may affect the viscoelastic properties of tendon and increase the risk of spontaneous rupture. PMID- 16470455 TI - Osteoarticular allograft of the proximal humerus--histopathological study 18 years after implantation. PMID- 16470456 TI - Idiopathic hypertrophy of the first dorsal interosseous and thenar muscles presenting as a tumor in a 12-year-old boy. PMID- 16470457 TI - Intrapelvic pin migration after periacetabular reconstruction and arthroplasty of the hip in metastatic pelvic disease--a case report. PMID- 16470458 TI - Total tibial endoprosthesis including ankle joint and knee joint replacement in a patient with Ewing sarcoma. PMID- 16470459 TI - No beneficial effects of joint distraction on early microscopical changes in osteoarthrotic knees. PMID- 16470461 TI - Selected complex auditory disorders. AB - This article provides an overview of four auditory disorders relevant generally to adults and especially to veterans. The disorders are noise-induced hearing loss, idiopathic sudden sensorineural hearing loss, otosclerosis, and Meniare's disease. Sensorineural hearing loss characterizes each, but additional aspects vary with each of the four conditions. This article describes the conditions, discusses their diagnoses and treatments, and outlines current and suggested rehabilitation. The emphasis is on recent advances, some of which await confirmation prior to possible acceptance as standard practice. PMID- 16470462 TI - Hearing loss and aging: new research findings and clinical implications. AB - This review provides an overview of recent research that addressed hearing loss and auditory processing problems among elderly people. It focuses on research from the University of Maryland on problems in auditory temporal processing by elderly listeners as assessed in speech perception experiments using temporally altered signals and in psycho-acoustic experiments of duration and rhythm discrimination for simple and complex signals. Some recent studies of perceived hearing disability are also reviewed. The clinical implications of the research findings are discussed in relation to hearing aid performance and use by elderly people as well as potential signal processing strategies that may prove to be beneficial for this population. PMID- 16470463 TI - Some effects of aging on central auditory processing. AB - Seniors often have more difficulty understanding speech than younger adults, particularly in noisy environments. While loss in peripheral hearing sensitivity explains many of the listening problems of elderly persons, age-related declines in general cognitive skill and central auditory processing also appear to contribute. In this article, we focus primarily on the effects of age on central auditory mechanisms. To this end, we review research examining a central locus for deficits in temporal processing and summarize behavioral and event-related potential findings from our laboratory's research on the effects of aging on dichotic listening performance. Results show that age-related deficits in interhemispheric information processing may underlie some of the listening problems among seniors. We also discuss implications for clinical audiological rehabilitative efforts in this population. PMID- 16470465 TI - Evaluation of human middle ear function via an acoustic power assessment. AB - Measurements of middle ear (ME) acoustic power flow (power reflectance, power absorption, and transmittance) and normalized impedance (acoustic resistance, acoustic reactance, and impedance magnitude) were compared for their utility in clinical applications. Transmittance, a measure of the acoustic power absorbed by the ME, was found to have several important advantages over other measures of acoustic power flow. In addition to its simple and audiologically relevant physical interpretation (absorbed power), the normal transmittance curve has a simple shape that is visually similar to the ME transfer function. The acoustic impedance measures (resistance and reactance) provided important additional information about ME status and supplemented transmittance measurements. Together these measurements can help identify unusual conditions such as eardrum perforations. While this article is largely a review of the development of a commercial power reflectance measurement system, previously unpublished experimental results are presented. PMID- 16470464 TI - Hearing health and care: the need for improved hearing loss prevention and hearing conservation practices. AB - Hearing loss affects 31 million Americans, particularly veterans who were exposed to harmful levels of noise during military functions. Many veterans also receive treatment with ototoxic medications, which may exacerbate preexisting hearing loss. Thus, hearing loss is the most common and tinnitus the third most common service-connected disability among veterans. Poor implementation of hearing protection programs and a lack of audiometric testing during medical treatment leave veterans vulnerable to unrecognized and untreated hearing loss until speech communication is impaired. Individualized audiometric testing techniques, including assessment of high frequencies, can be used in clinical and occupational settings to detect early hearing loss. Antioxidants also may alleviate cochlear damage caused by noise and ototoxicity. Ultimately, hearing loss prevention requires education on reducing occupational and recreational noise exposure and counseling on the risks and options available to patients. Technological advances will improve monitoring, allow better noise engineering controls, and lead to more effective hearing protection. PMID- 16470466 TI - Speech signals used to evaluate functional status of the auditory system. AB - This review presents a brief history of the evolution of speech audiometry from the 1800s to present day. The two-component aspect of hearing loss (audibility and distortion), which was formalized into a framework in past literature, is presented in the context of speech recognition. The differences between speech recognition in quiet and in background noise are discussed as they relate to listeners with normal hearing and listeners with hearing loss. A discussion of the use of sentence materials versus word materials for clinical use is included as is a discussion of the effects of presentation level on recognition performance in quiet and noise. Finally, the effects of age and hearing loss on speech recognition are considered. PMID- 16470467 TI - Clinical management of tinnitus using a "progressive intervention" approach. AB - Chronic tinnitus is experienced by 10%-15% of the population, of which only about 20% require clinical intervention. People requiring intervention have different levels of need, ranging from the provision of basic information to long-term, individualized treatment. We address this clinical need by outlining a five-level "progressive intervention" approach to the management of tinnitus that would provide a systematic framework for treatment by audiologists. At each level, patients must be appropriately referred-usually to otolaryngology, psychology, and/or psychiatry. Level 1 is an interview method of screening for determining if the person requires clinical intervention (and addressing basic questions). Level 2 is the provision of structured group educational counseling. If the screening determines that care is urgently required or if further help is needed following the group session(s), a tinnitus intake assessment (Level 3) should be performed. The intake assessment, which includes educational counseling, can often meet a patient's needs. If not, then a program of continuing treatment (Level 4) would be indicated. If significant benefit were not achieved through consistent treatment over 1-2 years, longer-term treatment (Level 5) would be indicated, which could include alternate or multiple treatment modalities. At all levels, the goal is to minimize the impact of tinnitus on the patient's life as efficiently as possible. PMID- 16470468 TI - Bilateral amplification and sound localization: then and now. AB - This article is concerned with the evolution and pros and cons of bilateral amplification. Determining whether a bilateral hearing aid fitting is superior to that of a monaural hearing aid is a long-standing question; for this reason, the trend toward bilateral amplification has been slow. However, it is now assumed that bilateral amplification has significant advantages over monaural amplification in most cases, a view that is supported by our localization results. In this article, we will address the advantages of bilateral hearing aids and reveal some new localization data that show that most listeners with bilateral amplification, when tested unaided, as well as normal-hearing listeners manifested very high degrees of symmetry in their judgments of perceived angle while listeners who routinely use monaural amplification and those with asymmetric hearing loss had relatively large asymmetries. These data show that asymmetry in localization judgments is a much more sensitive indicator of abnormal localization ability than the magnitude of localization errors. PMID- 16470469 TI - Directional hearing aids: then and now. AB - Directional microphone hearing aids can lead to improved speech recognition when speech and noise are coming from different directions. This technology provides limited benefits, however, and in specific instances use of a directional hearing aid mode can be detrimental. This article discusses the benefits and limitations of directional amplification, summarizes some current work in directional amplification, and recommends clinical application relative to the use of directional amplification. PMID- 16470470 TI - An evaluation of digital cellular handsets by hearing aid users. AB - Audible interference from digital cellular telephones (cell phones) has been a long standing problem for hearing aid users. The Federal Communications Commission (FCC) has lifted the hearing aid compatibility exception on cell phones and imposed a set of requirements effective September 2005. We conducted an experiment to determine how well hearing aid wearers are able to use commercially available digital cell phones. Hearing aid users evaluated the usability of six digital cellular handsets. The results suggest that certain transmission technologies create more annoyance from interference than others and that the type of hearing aid-to-telephone coupling (microphone or telecoil) can influence a user's experience of interference. However, the results also suggest that interference alone does not fully predict the usability of a cell phone for hearing aid users. These findings have implications for the American National Standards Institute C63.19 test and measurement standard that is used to rate cell phones' compliance with the FCC ruling and the education of consumers with regard to their expectations for cell phone use. PMID- 16470471 TI - Measuring hearing aid outcomes--not as easy as it seems. AB - Outcomes measurement in audiology has received much attention because of the need to demonstrate efficacy of treatment, provide evidence for third-party payment, carry out cost-benefit analyses, and justify resource allocation. Outcomes measurement shows the benefits obtained from a hearing aid and determines the costs of obtaining those benefits. In this article, we discuss why the seemingly simple issue of outcomes measurement is highly complex and the use of generic and disease-specific tools and the relationship between them; we also provide information regarding the International Classification of Functioning (ICF) system for selecting outcome measures. We then discuss factors complicating outcomes measurement, including discrepancies between clinically derived outcomes and functional outcomes, the ways clinicians can affect outcomes, and factors intrinsic to the patient that influence outcomes. We conclude that if the vision of moving quickly and efficiently from bench to chairside is to be realized, then clinicians must routinely measure hearing aid outcomes and researchers investigate their validity and usefulness. PMID- 16470472 TI - Central auditory system plasticity and aural rehabilitation of adults. AB - Until recently, researchers used behavioral measures of identification and discrimination of speech and nonspeech stimuli to assess the effects of auditory deprivation, enhancement, and training. Recent advances in our ability to measure electrical activity in the auditory system in response to sound have made it possible for us to study how changes in auditory input (because of hearing loss, auditory input modification, or training) affect the function of the central auditory system. This article reviews the evidence of changes in the auditory cortex in mature animals and in humans with acquired sensorineural hearing loss as well as changes associated with auditory training in persons with normal hearing. The results of studies that measure psychoacoustic and speech recognition performance of persons with hearing loss, with and without hearing aids, are interpreted within the framework of our new knowledge about plasticity of the auditory system. Applications of electrophysiologic techniques to hearing aid research and clinical practice are highlighted. PMID- 16470474 TI - The challenge for ultrasound in detecting lymph nodes and sentinel nodes. PMID- 16470473 TI - Hair cell regeneration: an exciting phenomenon...but will restoring hearing and balance be possible? AB - Sensory hair cells of the inner ear are susceptible to damage from a variety of sources including aging, genetic defects, and environmental stresses such as loud noises or chemotherapeutic drugs. Unfortunately, the consequence of this damage in humans is often permanent hearing/balance problems. The discovery that hair cells can regenerate in birds and other nonmammalian vertebrates has fueled a wide range of studies that are designed to find ways of restoring hearing and balance after such damage. In this review, we will discuss some of the key recent findings in sensory hair cell regeneration and what they mean for audiologists and other hearing healthcare practitioners. PMID- 16470475 TI - Application of transcranial sonography in extrapyramidal disorders: updated recommendations. AB - Transcranial sonography (TCS) has been proven to be a valuable tool in the diagnosis and differential diagnosis of extrapyramidal disorders, supplementing clinical and pathophysiological information that may be obtained by other neuroimaging techniques. Growing interest and increased application of TCS in extrapyramidal disorders require adherence to standardised principles concerning examination procedure and parameter settings of the ultrasound machine in order to ensure comparable results. This article provides an overview of these principles and typical findings in distinct extrapyramidal disorders. PMID- 16470476 TI - The current role of abdominal ultrasound in the clinical management of patients with AIDS. AB - Patients with AIDS present a wide variety of clinical manifestations through involvement of various organs. Ultrasonography (US) is easy to perform, safe, inexpensive, not invasive and repeatable. Those features are crucial for AIDS patients, who in industrialised countries are now mostly seen on an outpatient basis thanks to the introduction of Highly Active Antiretroviral Therapy. US can investigate most of the organs affected in AIDS and can guide biopsies, allowing the cyto-histological and microbiological investigations needed for a definitive diagnosis. This paper reviews the wide variety of applications of abdominal US and stresses its importance in the management of a complex and changing condition, particularly in settings where other more expensive imaging techniques are not--and will not be for a long time--available. The increasing use of portable/hand-carried scanners further adds to the value of the technique in such settings. With new treatments, prevalence and morbidity/mortality rates change, but new conditions and new side effects appear. US applications to these new conditions are discussed as well. PMID- 16470477 TI - Axillary sentinel lymph-node biopsy: gamma probe assisted sonographic localisation. AB - AIM: To evaluate axillary sentinel lymph-node (SLN) identification and localisation with gamma probe-assisted sonography. PATIENTS AND METHODS: The study comprises 42 patients. Sonographically guided peri-lesional injection of the radiopharmacon was performed on 40 breasts. Two patients with melanoma of the upper extremity underwent intracutaneous injection. After the acquisition of images with the gamma camera, the delineated lymph nodes were marked on the skin. A high-frequency transducer with an adapted gamma probe was then used to identify the SLN. Before surgery, the SLN was localised with a hook-wire. Node activity was controlled intraoperatively. RESULTS: In two out of 42 cases, lymphatic mapping did not reveal SLNs. Multiple SLNs at different sites, all with similar activity, were demonstrated in one patient, while in another patient the sonographically visible lymph nodes were too small for exact correlation. Hook wire localisation of the SLN was performed in the remaining 38 axillary spaces. Gamma probe-assisted sonography allowed for correct localisation of SLNs in 35/38 patients (92 %). Localisation was not correct in 3 patients (8 %), but in all cases the hook-wire was located next to the SLN. CONCLUSION: Preoperative identification and correct localisation of the SLN is possible in a high percentage of cases. This method allows rapid intraoperative detection of the SLN and could be used for exact correlation of sonographic appearance and histopathological results. PMID- 16470478 TI - Diagnostic accuracy of contrast-enhanced ultrasound in focal lesions of the liver using cadence contrast pulse sequencing. AB - The purpose of this study was to assess the accuracy of Cadence Contrast Pulse Sequencing (Siemens-Acuson, CA) method with injection of SonoVue (Bracco Imaging SpA, Italy) for the detection and characterisation of focal liver lesions in comparison with a reference modality during routine use. METHODS: 138 consecutive patients (62 women, 76 men; mean age: 52 years for women and 63 years for men) corresponding to 144 examinations with 381 lesions participated in this prospective study from December 2002 to November 2003. Inclusion criteria were lesions detected by conventional US and the availability of a reference imaging examination (CT or MR imaging) within three weeks. The underlying liver lesions characterised by a reference modality (including biopsy in 29 lesions) were distributed as follows: haemangioma (n = 56), focal nodular hyperplasia (n = 27), hepatocellular carcinoma (n = 44), hepatocellular adenoma (n = 5), liver metastasis (n = 174), abscess (n = 2), cysts (n = 45), other benign lesions (n = 24) and 3 peritoneal metastases. RESULTS: A significant improvement was reported in the number of detected lesions between contrast-enhanced ultrasound and baseline ultrasonography (351 lesions versus 280 lesions, respectively, p < 0.01), whereas no significant difference was noted between contrast-enhanced ultrasound and reference imaging (351 versus 377 lesions, respectively). On the whole, contrast-enhanced ultrasound allowed a complete diagnosis in 96 % of the detected nodules with a significant improvement compared to conventional sonography in which the diagnosis was suspected in only 52 % out of these cases (p < 0.001). No significant difference was noted between contrast-enhanced ultrasound and the reference modality concerning characterisation of nodules. CONCLUSION: The present study clearly indicates that contrast-enhanced sonography using Sonovue and Cadence Contrast Pulse Sequencing allows real-time imaging with high accuracy and thus will be a competitive alternative to other modalities such as CT and MR imaging for liver imaging. PMID- 16470479 TI - Initial experience with intraoperative ultrasound in navigated soft tissue operations of the neck and below the base of the skull. AB - AIM: Ultrasound (US) is a cost effective and time saving examination method for diseases of the neck,and can be used without any known side effects or limitations. US tied into modern navigation devices may significantly improve intraoperative orientation in regions with soft tissue characteristics. METHODS: 22 patients with soft tissue tumours of the head and neck underwent surgical procedures assisted by CAS system LandmarX (Medtronic) in combination with the US system DynaVievP II (Aloka). Clinical feasibility of using intraoperative US in navigated surgical procedures has been investigated by paying particular attention to the surgical approach, possible interferences from surgical instruments and time consumption. RESULTS: In the case of soft tissue shift, US can provide a sequence of a section of the operative field and, without delay, an instant comparison with the preoperative imaging data set. The feasibility of using this method is severely limited by the additional preparation required, the unfamiliar handling during surgery, and design-related matters. CONCLUSIONS: Integrating US into a navigation device could provide additional useful information for a more controlled resection of soft tissue disease of the neck and below the base of the skull. PMID- 16470480 TI - Parenchymal leiomyoma of the breast--clinical, sonographic, mammographic and histological features. AB - BACKGROUND: Intraparenchymal leiomyomas of the breast are quite rare. Areolar lesions are distinguished from intraparenchymal leiomyomas, which are less frequent. Clinically, leiomyomas appear as nodules; mammographically, they show up as round lesions. Reports on sonographic criteria are rare, and the criteria are nonspecific. Based on our case of an intraparenchymal leiomyoma, we describe additional sonographic features. The clinical, mammographic and sonographic characteristics of an intraparenchymal leiomyoma of the breast were evaluated. After surgery, the diagnosis was confirmed histologically. RESULTS: The clinical presentation of our patient with deep-seated leiomyoma of the breast included skin dimpling and a reduction in tissue mobility, differing from more commonly reported characteristics. Mammographically, the lesion was dense and only partly demarcated clearly, corresponding to other reports. On breast ultrasonography, the leiomyoma appeared as a hypodense, well demarcated, inhomogeneous lesion with posterior acoustic shadowing. A central tumour vessel was visible on Doppler imaging, and Cooper's ligaments were discontinuous. Acoustic shadowing, the hypodense character, hyperechoic border and the central tumour vessel are therefore additional ultrasonographic characteristics of an intraparenchymal leiomyoma of the breast. This type of lesion is usually described as isodense to hyperdense and homogeneous, possibly containing semicystic components. Previous reports have only described posterior acoustic enhancement, but not acoustic shadowing. CONCLUSION: On breast ultrasonography, an intraparenchymal leiomyoma of the breast can present with posterior acoustic shadowing, hypodense echogenicity, a hyperechoic border and a central tumour vessel. Neither imaging studies nor palpation allow distinction between benign and malignant lesions. PMID- 16470481 TI - [Unusual postoperative fistula of the abdominal wall, caused by an ingested wooden spit perforating the intestine and lodging itself in the abdominal wall]. AB - Abdominal fistula caused by an ingested wooden spit, which penetrated the intestinal wall and remained in the abdominal wall: a laparascopy was performed in a 41-year-old man suffering from acute appendicitis and an inflamed Meckel's diverticulum. After removal of the appendix and the diverticulum, a fistula developed in the excision channel of the left quadrant of the abdominal wall post operatively. Despite repeated incision of the abscess in the course of 2 months, the fistula did not heal. Ultrasound examination of the abdominal wall was therefore performed. The postoperative status was without conspicuous findings. We could, however, detect a foreign body, a few centimetres long and 2-3 mm thick, displaying a smooth surface, deep down in the abdominal wall. A fistulography confirmed the diagnosis. After removal of the wooden spit, complete healing of the fistula in the abdominal wall was observed. Subsequently, the patient reported to have eaten a beef roulade, fixed with a wooden spit, 7 weeks before the abdominal operation. PMID- 16470482 TI - The launch of the Renin Academy. AB - The Renin Academy was launched during the first week of September in Stockholm, at the European Society of Cardiology (ESC) annual congress. The Academy aims to encourage understanding of the renin-angiotensin-aldosterone system (RAAS) and to raise awareness of the benefits of optimising RAAS suppression. PMID- 16470483 TI - ASCOT a four horse race. PMID- 16470484 TI - Preventing end-stage renal disease in diabetic patients - dual blockade of the renin-angiotensin system (Part II). AB - Diabetic nephropathy is a major cause of diabetes related morbidity and mortality. The first part of the current review was published in the last issue of this journal and discussed the important role of the renin-angiotensin system (RAS) in diabetic nephropathy and the genetic influence on development of endstage renal disease (ESRD) in diabetic patients. This second part of the review focus on the potential improvement of the current treatment strategy to slow down the loss of kidney function using dual blockade of the RAS with both ACE-inhibitors (ACE-I) and angiotensin II receptor blockers (ARBs). Substantial evidence from short-term studies using surrogate endpoints indicates a beneficial impact of dual blockade of the RAS, not obtainable with single agent blockade alone, both in diabetic and non-diabetic renal disease. This conclusion has been confirmed and extended in a longterm trial with regard to prevention of ESRD in non-diabetic renal disease. Results indicate that dual blockade of the RAS may further slow down, but not arrest progressive loss of renal function. However, studies defining the optimal dose of ACE-I / ARBs without additional adverse effects are essential to ensure relevant comparison with dual blockade therapy. Trials using primary renal endpoints in diabetic nephropathy are still needed, and will finally establish the role of dual blockade of the RAS in a clinical setting. PMID- 16470485 TI - Collagen production in cardiac fibroblasts during inhibition of aminopeptidase B. AB - OBJECTIVE: To determine whether the aminopeptidase B inhibitor, arphamenine A, could affect collagen production and expression in control and TGF-ss1-treated cardiac fibroblasts. DESIGN AND METHODS: Cardiac fibroblasts from passage 2 from normal male adult rats were cultured to confluency and incubated with and without 600 pmol/l TGF-ss1 for 2 days in serum-free Dulbecco's modified Eagle's medium and then incubated with 100 mol/l arphamenine A for 1 day in this medium with added ascorbic acid, ss-aminopropionitrile and titriated proline. Soluble collagen was measured in the conditioned medium and non-soluble collagen in the cell layer. Aminopeptidase activity was estimated by spectrophotometric determination of the liberation of p-nitroaniline from alanine- or arginine-p nitroanilide. Matrix metalloproteinase (MMP) and lysyl oxidase activity were assayed in the conditioned medium. A semi-quantitative reverse transcriptase- polymerase chain reaction was used to examine the expression of lysyl oxidase and collagen type I and III. RESULTS: Arphamenine A dose-dependently inhibited basal and TGF-ss1-stimulated aminopeptidase activity. Arphamenine A reduced soluble and non-soluble collagen production in control and TGF-ss1-treated cardiac fibroblasts, while it decreased collagen type I and III expression only in TGF ss1-treated fibroblasts. Lysyl oxidase, MMP-1 and MMP-2 activity were inhibited by arphamenine A in the conditioned media of control and TGF-ss1-treated cardiac fibroblasts. CONCLUSIONS: Our data show that the specific aminopeptidase B inhibitor, arphamenine A, reduces collagen production in cardiac fibroblasts and that this reduction is accompanied by a pronounced inhibition of lysyl oxidase. PMID- 16470487 TI - Comparison of monotherapy with irbesartan 150 mg or amlodipine 5 mg for treatment of mild-to-moderate hypertension. AB - OBJECTIVE: The primary objective of this study was to compare the antihypertensive efficacy of the angiotensin II receptor blocker irbesartan 150 mg and the calcium channel blocker amlodipine 5 mg in the treatment of patients with seated diastolic blood pressure (DBP) 95-110 mmHg. DESIGN: Multicentre, randomised, double-blind, comparative pilot study. METHODS: Subjects were 18-65 years of age, with DBP 95-110 mmHg, and of non-African American origin. Following a three week, single-blind, placebo lead-in period, 181 subjects were randomised in a 1:1 ratio to receive once-daily irbesartan 150 mg (n=89) or amlodipine 5 mg (n=92) for four weeks. Trough (24+/-3 hours post-dosing) BP measurements were obtained at baseline and at Weeks 2 and 4 under standardised, controlled conditions. Response was defined as DBP <90 mmHg or a reduction from baseline of 10 mmHg. RESULTS: After four weeks of treatment, the mean (+/-SE) decrease from baseline in DBP was 9.4+/-0.6 mmHg in the irbesartan group vs. 9.6+/-0.6 mmHg in the amlodipine group (p=0.806). The mean decrease from baseline in seated systolic BP was 12.2+/-1.0 mmHg in the irbesartan group vs. 12.0+/-1.0 mmHg in the amlodipine group (p=0.885). Overall, 62% of subjects in the irbesartan group and 63% in the amlodipine group had a response (p=0.609), and 54% and 56% of patients (p=0.596), respectively, had their DBP normalised (<90 mmHg). Adverse events were reported by 21.3% of patients receiving irbesartan and 20.7% receiving amlodipine. Conclusions. Irbesartan 150 mg demonstrated comparable efficacy to amlodipine 5 mg, thereby confirming its value as an antihypertensive treatment option in non-African American patients with DBP 95-110 mmHg. PMID- 16470488 TI - Are multiple angiotensin receptor types involved in angiotensin (1-7) actions on isolated rat portal vein. AB - Angiotensin (1-7) [Ang (1-7)] is a bioactive component of the renin angiotensin system. Ang (1-7) may interact with angiotensin type 1 (AT1) or type 2 (AT2) receptors and with Ang (1-7) - specific receptors. We examined the interactions between different doses of Ang (1-7) (1 nM-1 microM) and angiotensin II (Ang II) (10 and 100 nM) on isolated rat portal vein. In endothelium-denuded portal vein rings, Ang (1-7) inhibited contractile effects induced by Ang II. The effects of Ang (1-7) were modified by indomethacin, N(G)-nitro-L-arginine methyl ester (L NAME), (D-Ala7)-Angiotensin (1-7) (H-2888) and losartan. Our results suggest that on rat isolated portal vein rings without endothelium, Ang (1-7) reduces Ang II induced contractions by acting mostly on Ang (1-7) specific receptors, and this effect is mediated by vasodilatatory prostaglandins. At high concentrations, Ang (1-7) effects are mediated by AT1-receptors, though to a lesser extent than by Ang (1-7) specific receptors. PMID- 16470486 TI - Renin release in response to Renin system blockade: activation of the Renin system in type 1 diabetes mellitus. AB - Activation of the renin-angiotensin system (RAS) in diabetes is thought to contribute to nephropathy. This is suggested by findings of an enhanced renovascular (RPF) response to RAS blockade with angiotensin-converting enzyme inhibitors (ACE-I) and angiotensin receptor blockers (ARBs). An alternative approach to assess RAS activation is the evaluation of renin release following RAS blockade. Forty-four consecutively enrolled Type 1 diabetic patients (28.2+/ 1.5 years) and 37 normal subjects (37+/-2.6 years) in high salt balance were given 25 mg of captopril and 16 mg of candesartan p.o. on consecutive days. All subjects were Caucasian. All, except one diabetic patient, had normal renal function. Plasma renin activity (PRA) and renal plasma flow (RPF) were measured for four hours after both drugs, and at eight, and 24 hours after candesartan. As anticipated, both drugs increased PRA. Peak responses (90' after captopril) were 5.6+/-1 ng/mL Ang I/hour in diabetic patients, and 1.7+/-0.9 ng/mL Ang I/hour in normal subjects (p<0.001). Responses to both drugs were correlated in diabetic patients for PRA (r=0.623; p=0.001) and for RPF (r=0.9; p<0.001). When the PRA response to captopril was below the median, the RPF response was limited (22.1+/ 17.6 ml/minute/1.73 m2). When it was above the median, the RPF response was also larger (62.2+/-13.9 ml/minute/1.73 m2; p=0.006). Renin response to ACE-I and ARB confirms activation of the RAS in diabetic patients. PMID- 16470490 TI - Effects of intravenous PD 123319 on haemodynamic and arterial stiffness indices in healthy volunteers. AB - Relatively little is known about the functional expression of cardiovascular angiotensin type 2 (AT2)-receptors in healthy young adult humans. We performed a randomised, placebo-controlled crossover study of the effects of intravenous administration of the selective AT2-receptor antagonist PD 123319 on haemodynamics and arterial stiffness in normal volunteers. Sixteen normal subjects aged 29.9+/-13.8 years (range 18-30 years) received an intravenous infusion of PD 123319 (10 mcg/minute for 5 minutes) and placebo, separated by one week. Haemodynamics (cardiac index, stroke index and systemic vascular resistance) were measured non-invasively using a BioZ.com thoracic impedance detection system. Blood pressure was measured from an arm cuff using oscillometry. Stiffness index, a measure of arterial stiffness, was measured using a Pulse Trace recorder. No significant changes in blood pressure (p=0.92), cardiac index (p=0.52), stroke index (p=0.61), systemic vascular resistance index (p=0.32) or stiffness index (p=0.57) was demonstrated following PD 123319 infusion, compared with placebo. The results of this study do not support the functional presence of cardiovascular AT2-receptors that mediate acute haemodynamic effects in healthy young adults. It remains possible that higher doses of PD 123319 may be required to demonstrate functional cardiovascular AT2 receptors in this population, if they are present. PMID- 16470489 TI - Does angiotensin (1-7) contribute to the anti-proteinuric effect of ACE inhibitors. AB - Angiotensin-converting enzyme inhibitors (ACE-I) reduce proteinuria and protect the kidney in proteinuric renal disease. During ACE-I therapy, circulating levels of angiotensin (1-7) [Ang (1-7)] are increased. As cardiac and renal protective effects of Ang (1-7) have been reported, we questioned whether Ang (1-7) contributes to the anti-proteinuric effects of ACE-I treatment. Therefore, we evaluated whether Ang (1-7) infusion reduces proteinuria in a rat model of adriamycin-induced renal disease. In addition, the effect of a selective Ang (1 7) blocker, [D-Ala7]-Ang (1-7) (A779), was investigated in rats treated with the ACE-I, lisinopril (LIS). Six weeks after induction of proteinuria, therapy was started in four different groups: control, Ang (1-7), LIS, and LIS+A779. After two weeks, the rats were sacrificed. Six weeks after injection of adriamycin, the rats had developed proteinuria of 323+/-40 mg/24 hours. The proteinuria remained stable in the control group and in the Ang (1-7) group, but was reduced in both LIS and LIS+A779-treated groups. Similarly, blood pressure (BP) was unchanged in the control and the Ang (1-7) groups, but reduced in both the LIS and the LIS+A779 groups. Plasma levels of Ang (1-7) were increased in the Ang (1-7) and in both LIS-treated groups. We conclude that systemic Ang (1-7) plays no major role in the anti-proteinuric and BPlowering effects of ACE-I in this rat model of adriamycin-induced nephrosis. PMID- 16470491 TI - Renin report: spotlight on Renin: therapeutic opportunities for Renin inhibitors. PMID- 16470495 TI - Female prisoners with borderline personality disorder: some promising treatment developments. AB - BACKGROUND: The over-representation of female prisoners with borderline personality disorder (BPD) is an area of concern for HM Prison Service. Pilot programmes of Dialectical Behaviour Therapy (DBT) were undertaken for the first time in three British prisons for women diagnosed with BPD. Standard year-long programmes were piloted in two closed training prisons. Three short-format programmes were undertaken in a local allocation prison. METHOD: Evaluation measures included psychometric tests, behavioural data, and interviews with participants and key personnel. Sixteen of the 30 women who embarked on the programmes completed them, though five dropouts were transferred or released, leaving a voluntary attrition rate of 33%. Fourteen completed all measures. A waiting-list control group of eight participants was also set up. Five completed all measures. RESULTS: The vast majority of completers showed overall improvements in psychometric data often reaching statistical significance, and with notable effect sizes, while there was no significant overall change in the control group (though improvements were seen). A down-turn in overall self-harm was also seen. CONCLUSION: Results are tentative at this stage because of the small sample size. However, despite the numerous challenges associated with implementation, outcomes showed real promise for delivering DBT in a prison setting, and its efficacy in reducing criminogenic risk and improving the manageability and quality of life for this highly problematic group. Lessons learned for future implementation in correctional settings are discussed. PMID- 16470496 TI - The manualization of a treatment programme for personality disorder. AB - BACKGROUND: The advantages of manualized psychological treatments include: the promotion of evidence-based practice, the enhancement of treatment integrity, the facilitation of staff training, and the potential replicability of treatment. ARGUMENT: The manualization of a multi-component, multidisciplinary treatment programme for male personality-disordered offenders is described. The background to this development is explained and the treatment setting is described briefly, followed by a description of the eight treatment manuals: (1) the treatment overview, (2) Psychoeducation focusing on personality disorder diagnosis and core beliefs, (3) Trust and Self-awareness group exercises, (4) Stop and Think! -- a social problem-solving intervention, (5) Controlling Angry Aggression, (6) Controlling Substance Use, (7) Criminal Thinking/Belief Therapy, and (8) Skills for Living -- a social skills manual. CONCLUSIONS: In addition to the original aims of manualization, this exercise has clarified the treatment programme, included less highly trained staff in the delivery of therapy and permitted the evaluation of treatment modules, thus contributing to the incremental evaluation of the overall programme. These manuals may usefully be shared with other practitioners in the field. PMID- 16470497 TI - Using the SWAP-200 in a personality-disordered forensic population: is it valid, reliable and useful? AB - BACKGROUND: Treatment and risk management of forensic patients relies heavily on diagnosing psychopathology, yet the reliability of clinical diagnoses of personality disorder has been found to be only fair to low. Structured instruments for the global assessment of personality disorder are infrequently used in clinical assessments possibly due to their limited validity and clinical utility. AIMS/METHODS: The Shedler-Westen Assessment Procedure-200 (SWAP-200) was developed in an effort to address these limitations. Although good reliability and validity in relation to clinicians' diagnosis of personality disorder has been reported, to date the validity of this instrument has not been assessed in relation to other standardized instruments or in a personality-disordered, forensic population. This study aims to establish the reliability and validity of the SWAP-200 against other diagnostic instruments and measures of interpersonal functioning in a personality disordered forensic population. RESULTS: This paper reports the results of 30 subjects from a high secure hospital in the UK who were assessed with the SWAP-200, the Structured Clinical Interview for DSM-IV Personality Disorders (SCID-II), the Adult Attachment Interview (AAI) and the Chart of Interpersonal Reactions in Closed Living Environments (CIRCLE). Preliminary results suggest that the SWAP-200 is a reliable instrument for the diagnosis of personality disorder in forensic patients. CONCLUSIONS: Although the small sample size allows only preliminary conclusions about the validity of this instrument, early results show a reduction of the diagnosis of comorbidity compared with the SCID-II, together with an increased number of expected associations between independent measures of interpersonal functioning and categories of personality disorder. PMID- 16470498 TI - Protective factors for youth considered at risk of criminal behaviour: does participation in extracurricular activities help? AB - BACKGROUND: There is a lack of research investigating the potential protective effect of participation in extracurricular activities on youth who are at risk of engaging in delinquent activity. AIM: This study examined the potential for participation in extracurricular activities to act as a protective factor for youth deemed at risk of engaging in delinquent activity. METHOD: One hundred and sixty-nine secondary students from Glasgow, Scotland completed two questionnaires (the Youth Self-Report and an additional information sheet) requesting information about their participation in extracurricular and delinquent activities as well as their possible risk factors. Activities included sports, non-sports (hobbies and games), current activities (youth clubs and other organisations) and previous involvement in activities. Risk factors included residing in a broken home, having four or more siblings, academic failure and lacking a non-parental very important person. Delinquent activities included rule breaking and aggressive behaviours. RESULTS: Independent samples t-tests found that females participated in significantly more non-sports and previous activities than males and that males participated in significantly more rule breaking behaviour than females. Hierarchical multiple regression analyses found that gender and participation in sports were strong predictors of rule-breaking behaviour. A significant positive correlation was found between participation in sports and involvement in aggressive behaviour. CONCLUSION: The results suggest that participation in extracurricular activities does not act as a protective factor for youth, regardless of whether or not they are considered to be at risk of engaging in delinquent activity. The significant correlation found between participation in sports and involvement in aggressive behaviour suggests that youth participation in sports may act as a risk factor. PMID- 16470499 TI - Is gender a factor in perceived prison officer competence? Male prisoners' perceptions in an English dispersal prison. AB - BACKGROUND: The introduction of women officers into HM Prison Service raised questions regarding women's ability to perform what had traditionally been a male role. Existing research is inconclusive as to whether female prison officers are as competent as male prison officers, and whether there are gender differences in job performance. This study examined prisoners' perceptions of male and female prison officers' performance. HYPOTHESES: The hypotheses were that overall competence and professionalism ratings would not differ for men and women officers, but that there would be differences in how men and women were perceived to perform their roles. Women were expected to be rated as more communicative, more empathic and less disciplining. METHOD: The Prison Officer Competency Rating Scale (PORS) was designed for this study. Ratings on the PORS for male and female officers were given by 57 adult male prisoners. RESULTS: There was no significant difference in prisoners' ratings of overall competence of men and women officers. Of the PORS subscales, there were no gender differences in Discipline and Control, Communication or Empathy, but there was a significant difference in Professionalism, where prisoners rated women as more professional. CONCLUSION: The failure to find any differences between men and women in overall job competence, or on communication, empathy and discipline, as perceived by prisoners, suggests that men and women may be performing their jobs similarly in many respects. Women were rated as more professional, and items contributing to this scale related to respecting privacy and keeping calm in difficult situations, where there may be inherent gender biases. PMID- 16470500 TI - Victim services in the National Health Service (NHS): combining treatment with violence prevention. PMID- 16470501 TI - Mental retardation and the death penalty in the USA: the clinical and legal legacy. PMID- 16470502 TI - Temporal reliability of psychological assessments for patients in a special hospital with severe personality disorder: a preliminary note. AB - BACKGROUND: The new programme for assessing those with dangerous and severe personality disorder relies heavily on psychological assessments of personality disorder and risk. METHODS: The temporal reliability of assessments of psychopathy (PCL-R), risk (HCR-20) and personality was assessed using the International Personality Disorder Examination (IPDE) in 15 randomly selected male prisoners in a high secure hospital carried out at intervals varying between a mean of nine and 19 months after initial assessments by a variety of assessors. RESULTS: Using the intra-class correlation coefficient the agreement varied between 0.57 (HCR-20), 0.58 (PCL-R) and 0.38-0.70 for IPDE personality disorders, with the best agreement for antisocial personality disorder (0.70). COMMENT: These levels of agreement are consistent with other recent work on temporal reliability of personality instruments but are a little too low for confidence in these measures alone in the assessment process. PMID- 16470503 TI - Mortality among mentally disordered offenders: a community based follow-up study. AB - BACKGROUND: Follow-up information about outcome for hospitalized mentally disordered offenders (MDO) is necessary for evaluation and improvement in quality of forensic psychiatric care. AIM: A study was undertaken to estimate the standard mortality rate (SMR) of a population based sample of people sentenced to forensic psychiatric care. METHOD: All MDOs in Orebro County, Sweden, discharged from a forensic psychiatric treatment unit between 1992 and 1999 were identified (n = 46). The variables were gender, age, offence, diagnosis and duration of admission. Case linkage was made with the National Cause-of-Death register. Median follow-up time was 53 months (0-93). RESULTS: The sample yielded a significantly elevated SMR 13.4 (95% CI 4.35-31.3) times higher than that in the general population, mostly due to suicide. CONCLUSIONS: The cohort size is small but representative, and it provides data from an additional country for the growing international pool confirming the high risk of premature, generally self inflicted death among MDOs. Resettlement and rehabilitation services for them may need to take as much account of mortality risk as that of re-offending. PMID- 16470504 TI - Animal abuse among preadolescents directly and indirectly victimized at school and at home. AB - BACKGROUND: Animal abuse by preadolescents has been associated with their later family violence and/or criminal behaviour; less is known about animal abuse and concurrent experience of being a victim at home and/or school, or of contemporaneous aggression to peers. AIMS: To establish the prevalence of animal abuse among Italian preadolescents and its relationship with experience of abuse at home and school (direct and witnessed), and to peer abuse (bullying). METHOD: An Italian community sample of 268 girls and 264 boys (aged 9-12) completed a self-reported questionnaire about victimization at home and school, animal abuse and bullying. RESULTS: Two in five preadolescents admitted abusing animals at least once in their life, and one in three bullying peers at school, with a higher prevalence among boys. Over three-quarters of all participants reported at least one type of victim experience: one-third had experienced inter-parental violence; over one-third had themselves been abused by one or both parents; two in five had been directly or indirectly victimized at school. Individual tests of association suggested gender differences. Multivariate regression analyses conducted separately for boys and girls showed that the independent variable accounted for more than 25% of the variance for the girls, but less than 10% for the boys. Experiences of abuse were the key independent variables for the girls; other expressions of aggression were the key variables for the boys. DISCUSSION AND CONCLUSIONS: The results suggest that discovery of animal abuse should prompt further enquiries about other problems that a child may have. Detection of animal abuse by a child could offer an early opportunity for intervention to alleviate internalized damage or other aggressive behaviour. PMID- 16470505 TI - Does the non-randomized controlled study have a place in the systematic review? A pilot study. AB - BACKGROUND: A major issue in any systematic review is deciding which trials or studies to include and which to exclude. The Cochrane Collaboration and similar respected organizations have traditionally viewed the randomized trial (RCT) as the only acceptable evidence on treatment outcome. However, many systematic reviews are indeterminate because they include insufficient RCTs whilst they reject large numbers of non-randomized controlled studies. This is particularly true in forensic mental health, a domain where RCT methodology can be problematic. Systematic reviews could become more informative if reviewers knew when, and under what circumstances, non-randomized designs are acceptable for inclusion alongside RCTs. METHOD: This pilot study explores whether good-quality, controlled, non-randomized studies can be reliable surrogates for RCTs. We examined two published reviews from the Cochrane Schizophrenia Group. We compared outcomes between (a) randomized trials (that had been included) and non randomized studies (that had been excluded), and (b) between high- and low quality studies using an established quality checklist. RESULTS: In the first review, effect scores were similar for randomized and non-randomized studies and relatively insensitive to study quality. In the second review, the treatment effect was considerably lower for the RCT group - here, however, studies of high quality showed much smaller effect scores than those of low-quality on two separate outcomes. CONCLUSIONS: Non-randomized controlled studies of high quality can produce outcomes that approximate to those found in RCTs. Trial quality may have a greater impact on treatment effect size than randomization alone, suggesting that randomization should not be seen as a reliable proxy for overall quality. The problems and issues still to be resolved are discussed with recommendations for future research. PMID- 16470506 TI - Can non-randomized trials be considered to be equivalent to randomized trials in forensic mental health? PMID- 16470507 TI - Can we obtain credible estimates of the causal effects of interventions from non randomized trials (observational studies or so-called quasi-experiments)? PMID- 16470508 TI - Predictors of contact difficulty and refusal in a longitudinal study. AB - BACKGROUND: Attrition presents a serious problem to researchers collecting longitudinal data. Participant loss threatens both the internal and external validity of research findings. This study sought to examine predictors related to contact difficulty and refusals in a longitudinal study. METHOD: Data for this paper came from the Developmental Trends Study, a longitudinal study investigating the development of disruptive behaviour disorders in a sample of 177 clinic-referred boys. Annual follow-up assessments were conducted, ages for the periods examined ranged from 11 to 19 years. The predictor domains during project years 1-4 included demographics, child functioning, parental functioning, parenting skills and participant dispersion. RESULTS: These indicated that participant's older age, low socioeconomic status, the presence of callous and un emotional behaviours, and having a father with antisocial personality disorder predicted contact difficulty, while participant's older age, living in a rural environment and attention deficit hyperactivity disorder were predictive of refusals. Participant dispersion was not significantly related to either contact difficulty or refusal status. CONCLUSIONS: Investigating a broader range of variables may better allow researchers to identify participants who may be at risk of attrition. Early identification of 'difficult' participants enables development of retention strategies to minimize attrition. PMID- 16470510 TI - Ultra-performance liquid chromatography coupled to time-of-flight mass spectrometry for robust, high-throughput quantitative analysis of an automated metabolic stability assay, with simultaneous determination of metabolic data. AB - The application of ultra-performance liquid chromatography coupled to time-of flight mass spectrometry (UPLC/TOFMS) for high-throughput analysis of a 96-well plate based metabolic stability assay has been investigated. Full-scan data were acquired, with run times of 2.5-3.5 min, from which narrow window extracted ion chromatograms were generated, producing quantitative data for the test compound equivalent to that obtained by high-performance liquid chromatography with tandem mass spectrometric detection on a triple quadrupole instrument (HPLC/MS/MS). Sensitivity and mass accuracy were maintained over the analysis of >300 samples. Additionally, the UPLC/TOFMS datasets obtained gave access to metabolic route information, at no cost in terms of sensitivity for the test compound. PMID- 16470511 TI - Molecular cytogenetic analysis of de novo dup(5)(q33.1q31.1) associated with abnormal perinatal findings. PMID- 16470512 TI - Prenatal diagnosis of pure distal 18q deletion. PMID- 16470513 TI - Simultaneous quantification of atorvastatin and active metabolites in human plasma by liquid chromatography-tandem mass spectrometry using rosuvastatin as internal standard. AB - A simple, sensitive, selective and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of atorvastatin and its active metabolites ortho-hydroxyatorvastatin and para hydroxyatorvastatin in human plasma using rosuvastatin as internal standard (IS). Following simple liquid-liquid extraction, the analytes were separated using an isocratic mobile phase on a reversed-phase C18 column and analyzed by MS in the multiple reaction monitoring mode using the respective [M+H]+ ions, m/z 559/440 for atorvastatin, m/z 575/466 for ortho-hydroxyatorvastatin, m/z 575/440 for para hydroxyatorvastatin and m/z 482/258 for the IS. The assay exhibited a linear dynamic range of 0.1-20 ng/mL for atorvastatin and its two metabolites in human plasma. The lower limit of quantification was 100 pg/mL with a relative standard deviation of less than 8%. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. The average absolute recoveries of atorvastatin, ortho-hydroxyatorvastatin, para-hydroxyatorvastatin and the IS from spiked plasma samples were 54.2 +/- 3.2, 50.1 +/- 3.8, 65.2 +/- 3.6 and 71.7 +/- 2.7%, respectively. A run time of 2.5 min for each sample made it possible to analyze more than 300 human plasma samples per day. The validated method has been successfully used to analyze human plasma samples for application in pharmacokinetic, bioavailability or bioequivalence studies. PMID- 16470514 TI - The effect of temperature on the analysis of metanephrine for catechol-O methyltransferase activity assay by HPLC with electrochemical detection. AB - The enzyme catechol-O-methyltransferase (COMT) plays an important role in the metabolism of catechol estrogens and degradation of the catecholamine neurotransmitters, such as epinephrine. Several analytical methods, mainly high performance liquid chromatography with electrochemical amperometric detection, have been reported for the analysis of catecholamines and their metabolites in biological fluids. In this paper we report the relevance of controlling temperature in calibration procedures of metanephrine, an O-methylated product of catechol-O-methyltransferase, using epinephrine as substrate. The results at higher temperatures show shorter retention times of metanephrine, no undue band broadening and increased electro signals. This study also showed that, despite different temperatures leading to similarly specific activities of recombinant human COMT as expected, there are additional advantages in flow analytical methods where good sensitivity, efficiency and selectivity is required, mainly in tissues with low levels of COMT activity. PMID- 16470515 TI - Purification of a crystallin domain of Yersinia crystallin from inclusion bodies and its comparison to native protein from the soluble fraction. AB - It has been established that many heterologously produced proteins in E. coli accumulate as insoluble inclusion bodies. Methods for protein recovery from inclusion bodies involve solubilization using chemical denaturants such as urea and guanidine hydrochloride, followed by removal of denaturant from the solution to allow the protein to refold. In this work, we applied on-column refolding and purification to the second crystallin domain D2 of Yersinia crystallin isolated from inclusion bodies. We also purified the protein from the soluble fraction (without using any denaturant) to compare the biophysical properties and conformation, although the yield was poor. On-column refolding method allows rapid removal of denaturant and refolding at high protein concentration, which is a limitation in traditionally used methods of dialysis or dilution. We were also able to develop methods to remove the co-eluting nucleic acids during chromatography from the protein preparation. Using this protocol, we were able to rapidly refold and purify the crystallin domain using a two-step process with high yield. We used biophysical techniques to compare the conformation and calcium-binding properties of the protein isolated from the soluble fraction and inclusion bodies. PMID- 16470516 TI - A novel 14C-postlabeling assay using accelerator mass spectrometry for the detection of O6-methyldeoxy-guanosine adducts. AB - Accelerator mass spectrometry (AMS) is currently one of the most sensitive methods available for the trace detection of DNA adducts and is particularly valuable for measuring adducts in humans or animal models. However, the standard approach requires administration of a radiolabeled compound. As an alternative, we have developed a preliminary 14C-postlabeling assay for detection of the highly mutagenic O6-methyldeoxyguanosine (O6-MedG), by AMS. Procedures were developed for derivatising O6-MedG using unlabeled acetic anhydride. Using conventional liquid chromatography/mass spectrometry (LC/MS) analysis, the limit of detection (LOD) for the major product, triacetylated O6-MedG, was 10 fmol. On reaction of O6-MedG with 14C-acetic anhydride, using a specially designed enclosed system, the predominant product was 14C-di-acetyl O6-MedG. This change in reaction profile was due to a modification of the reaction procedure, introduced as a necessary safety precaution. The LOD for 14C-di-acetyl O6-MedG by AMS was determined as 79 amol, approximately 18,000-fold lower than that achievable by liquid scintillation counting (LSC). Although the assay has so far only been carried out with labeled standards, the degree of sensitivity obtained illustrates the potential of this assay for measuring O6-MedG levels in humans. PMID- 16470517 TI - Analysis of amitrole by normal-phase liquid chromatography and tandem mass spectrometry using a sheath liquid electrospray interface. AB - The coupling of normal-phase liquid chromatography to tandem mass spectrometry, previously developed in our laboratory, has been applied to the analysis of amitrole. This coupling utilizes an electrospray interface modified to accommodate the introduction of a make-up solution at the tip of the electrospray probe. A methanolic solution containing 3 mM ammonium acetate delivered at a flow rate of 10 microL . min(-1) was found to be the optimal sheath liquid to promote successful ionization of the amitrole. Protonated molecules, arising from in source dissociation of ammonium adducts, were subjected to tandem mass spectrometric experiments in a triple-quadrupole instrument. The main fragmentation reactions were characterized and selected to acquire chromatographic data in the multiple reaction monitoring mode. The limit of detection for amitrole was in the ppm range without any preconcentration step. Enhanced efficiency of ion transmission achievable nowadays in mass spectrometers (this analytical configuration was developed with a 15-year-old instrument) is reasonably expected to further improve this detection level. PMID- 16470518 TI - Brain imaging evidence of preclinical Alzheimer's disease in normal aging. AB - OBJECTIVE: This study was designed to test the hypothesis that baseline glucose metabolism and medial temporal lobe brain volumes are predictive of cognitive decline in normal older people. METHODS: We performed positron emission tomography using [18F]fluorodeoxyglucose and structural magnetic resonance imaging at baseline in 60 cognitively normal community-dwelling older subjects who were part of a longitudinal cohort study. Subjects were followed for a mean of 3.8 years, with approximately annual evaluation of global cognition (the Modified Mini-Mental State Examination) and episodic memory (delayed recall). Baseline brain volumes and glucose metabolism were evaluated in relation to the rate of change in cognitive test scores. RESULTS: Six subjects developed incident dementia or cognitive impairment (converters). Baseline positron emission tomography scans showed regions in left and right angular gyrus, left mid temporal gyrus, and left middle frontal gyrus that predicted the rate of change on the Modified Mini-Mental State Examination (p < 0.001). The left hemisphere temporal and parietal regions remained significant when converters were excluded. Both hippocampal (p = 0.03) and entorhinal cortical volumes (p = 0.01) predicted decline on delayed recall over time, and entorhinal cortical volumes remained significant when converters were excluded (p = 0.02). These brain volumes did not predict Modified Mini-Mental State Examination decline. INTERPRETATION: These results indicate that temporal and parietal glucose metabolism predict decline in global cognitive function, and medial temporal brain volumes predict memory decline in normal older people. The anatomical location of these findings suggests detection of preclinical Alzheimer's disease pathology. PMID- 16470519 TI - Efficacy of botulinum-A toxin bladder injections for the treatment of neurogenic detrusor overactivity in multiple sclerosis patients: an objective and subjective analysis. AB - AIMS: We studied the use of botulinum-a toxin (BTX-A) injections into the bladder as an alternative approach in patients with neurogenic detrusor overactivity due to multiple sclerosis (MS) with drug-refractory overactive bladder (OAB) symptoms. METHODS: Sixteen MS patients--11 women, 5 men; mean age 48.6 years- with refractory OAB symptoms were included in a one-center prospective study. For outcome analysis, we used a bladder diary, a complete urodynamic study, and validated questionnaires for subjective assessment. We injected 300 U of BTX-A (Botox) into the bladder and into the external sphincter muscle to reduce the probability of posttreatment urine retention. RESULTS: There was an increase in residual volume from 81.3 +/- 23.8 to 126.3 +/- 32.9 ml after 4 weeks. In one woman, transient self-catheterization was unavoidable. Four weeks and 3 and 6 months after BTX-A injection, the significant results were as follows: daytime frequency was reduced by 29%, 44%, and 30%, respectively. Nocturia diminished by 33%, 72%, and 40%. Use of pads was be reduced by 38% after 4 weeks and by 64% after 3 months. Urodynamically, reflex volume and maximal cystometric bladder capacity increased by 73%, 77%, and 58% (at 6 months, the increase was not significant) and by 36%, 27%, and 36% (not significant). Maximal detrusor pressure decreased by 35%, 22%, and 57%. Subjective outcome indicated significant improvement of symptoms at 4 weeks and 3 months, but not at 6 months. Patient satisfaction with the therapy was very high. CONCLUSIONS: BTX-A detrusor injections are very effective in the treatment of drug-resistant OAB symptoms in MS patients as reflected in urodynamic measurements and in patient satisfaction. Build up of residual urine remains a problem of which patients must be informed. PMID- 16470520 TI - Adolescents with acute lymphoblastic leukaemia: emerging from the shadow of paediatric and adult treatment protocols. AB - Adolescents and young adults (AYA) with acute lymphoblastic leukaemia (ALL) constitute a distinct population from children and older adults. Based on patterns of referral, they may be treated by either paediatric or adult oncologists. As a group, AYA with ALL have a worse survival and event-free survival (EFS) compared to that achieved by younger children. A systematic review of all published clinical trials, which provide data on treatment and outcome of adolescents with ALL, has been summarised in an effort to determine whether they should be treated on paediatric or adult type protocols. Adolescents appear to have a consistent survival advantage when treated on paediatric regimens. PMID- 16470521 TI - Central nervous system-related permanent consequences in patients with Langerhans cell histiocytosis. AB - BACKGROUND: Permanent consequences in Langerhans cell histiocytosis (LCH) are irreversible late sequelae related to the disease that may severely impair the quality of life of survivors. The frequency and pattern of permanent consequences affecting the central nervous system (CNS) remains to be determined. PROCEDURE: In this single center study, 25 LCH patients observed for a median time of 10 years 3 months underwent a uniform thorough follow-up program including neuropsychological testing and electrophysiological evaluation. RESULTS: Overall permanent consequences were seen in 9 of 25 patients. Intracranial abnormalities were the most frequent including diabetes insipidus (DI) in seven patients, anterior pituitary deficiencies in five patients, and neurodegenerative CNS disease in five patients. No patient had overt neurological symptoms upon neurological evaluation, but psychological testing revealed subtle deficits in short-term auditory memory (STAM) in 14 patients. Brain stem evoked potentials showed abnormalities in four of nine tested patients, all of these four had neurodegeneration on MRI. CONCLUSION: Psychoneuroendocrine sequelae were found in an unexpectedly high number of patients in this single center study. Long-term follow-up focusing on such sequelae are important in LCH survivors, in order to detect early deficits, to monitor the evolution of the disease, and to provide specific support. PMID- 16470522 TI - Regulated release of VWF and FVIII and the biologic implications. AB - von Willebrand factor (VWF) performs a critical function in platelet binding at the site of vascular injury and also serves as the carrier protein for coagulation factor FVIII (FVIII), protecting it from proteolytic degradation in plasma. Both proteins undergo rapid, regulated release in response to DDAVP administration in patients with mild hemophilia A or von Wille-brand disease. Here, we attempt to summarize our current understanding of the establishment of the regulated storage pool of VWF and FVIII. The data presented indicate that regulated secretion of both proteins occurs only if there is endogenous synthesis of FVIII together with VWF. PMID- 16470523 TI - DNA damage in bone marrow and blood cells of mice exposed to municipal sludge leachates. AB - Leachates of municipal solid waste from unsecured disposal sites contaminate sources of potable water and affect human health. In the present study, we have used the Comet assay to evaluate the DNA damage in mice exposed to municipal sludge leachates. Ten percent leachates were prepared from municipal sludge obtained from two different disposal drains. Male Swiss albino mice were treated daily with 0.1-0.4 ml of the leachates by oral gavage for 15 days, and the DNA damage was evaluated in bone marrow and blood using Olive tail moment, % tail DNA, and tail length as measures of DNA damage. Physicochemical and metal analysis of the leachates detected the presence of cadmium, chromium, copper, nickel, lead, and zinc, as well as elevated concentrations of sulfate and nitrate. Both of the leachates produced significant dose-responsive increases in DNA damage in both mouse tissues. There were no significant differences in the responses for any of the Comet endpoints between tissues (for the same leachate sample) or between leachate samples (for the same tissue). The results of this study indicate that municipal waste leachates produce DNA damage in vivo. PMID- 16470524 TI - Radiation-induced DNA damage and repair in peripheral blood mononuclear cells from Nijmegen breakage syndrome patients and carriers assessed by the Comet assay. AB - Nijmegen breakage syndrome (NBS) patients and carriers are predisposed to malignancy and are often treated with X-irradiation. In the present study, the single-cell gel electrophoresis (Comet) assay was used to examine radiation induced DNA damage and repair in peripheral blood mononuclear cells from NBS patients (n=13) and carriers (n=36) of six unrelated families. Cells from apparently healthy donors (n=10) and from breast cancer patients with normal clinical radiosensitivity (n=10) served as controls. Cells were irradiated with 5 Gy of X-rays and assayed for initial DNA damage and for residual DNA damage after 40 min of repair; the kinetics of DNA repair also was estimated. In addition, the nuclear area of unirradiated cells was extracted from the Comet data. The initial radiation-induced DNA fragmentation indicated that cells from members of two out of six NBS families were significantly more sensitive to X-irradiation than cells from the controls. Cells from four NBS families had longer DNA repair half-time values, while cells from five NBS families had significantly increased residual DNA damage following repair. The mean nuclear area of unirradiated cells processed in the Comet assay was 1.3-fold higher in cells from all NBS families than in the controls (P<0.05). Notably, the Comet assay parameters (initial and residual DNA damage and the repair kinetics) of irradiated NBS cells predicted the carrier status of the majority (86%) of blindly tested individuals. The prediction of NBS status was higher if the nuclear area of unirradiated cells was used as the endpoint. The results of this study suggest that the impaired radiation response of NBS cells should be taken into account if radiotherapy of NBS patients and carriers is required. PMID- 16470526 TI - Frequency-dependent conduction block in carpal tunnel syndrome. AB - Frequency-dependent conduction block (FDB) across segments of demyelination in response to high-frequency nerve stimulation has been well demonstrated in animals and has been explored in humans. However, attempts to demonstrate this phenomenon in sensory fibers involved in entrapment neuropathies have been unsuccessful. Therefore, we investigated the effects of high-frequency nerve stimulation in the median motor nerve in patients with carpal tunnel syndrome (CTS) with moderate to severely increased distal motor terminal latencies (MTL). As a group, the mean decrease in negative peak amplitude (npAmp) during 20 stimuli at 30-HZ frequency was significantly greater in CTS subjects (-11.3%) than in controls (+7.9%). The degree of FDB was greater when MTL was more prolonged (i.e., -4.9% at 5.0 ms and -25.3% at 9.4 ms) and FDB was more pronounced at higher stimulation frequencies (20 and 30 HZ). Our results suggest that the safety margin for impulse transmission is impaired in the motor axons of patients with a focal demyelinating lesion. These findings may correlate with the observation of weakness in the absence of conduction block in patients with entrapment neuropathies. PMID- 16470525 TI - Limited lung resection using the potassium-titanyl-phosphate laser. AB - BACKGROUND AND OBJECTIVES: Recently, the use of the potassium-titanyl-phosphate (KTP) laser with a wavelength of 532 nm, which is half the wavelength of the neodymium:yttrium-aluminum-garnet (Nd:YAG) laser, has been expanding in many fields, since this laser offers hemostasis and vaporization with minimal coagulation of the underlying structures. The purpose of this study was to determine the usefulness of the KTP laser in limited lung resection done for pulmonary nodular lesions. STUDY DESIGN/PATIENTS AND METHODS: We used the KTP laser to manage 10 partial resections (Group 1) and 16 segmentectomies (Group 2). The variables that were used to compare the two methods of resection included intra-operative blood loss, mean drainage time, and post-operative morbidity. RESULTS: The mean intra-operative blood loss was smaller in Group 1 (152.8 +/- 65.0 cc, P = 0.002) than in Group 2 (341.9 +/- 178.6 cc). Mean drainage time was shorter in Group 1 (1.8 +/- 1.0 days, P = 0.0141) than in Group 2 (3.6 +/- 2.9 days). The results of both groups with respect to hemostatic and healing effects were satisfactory. On histopathologic evaluation, the KTP laser-irradiated field tissue alteration consisted of two layers with a depth up to 2 mm. CONCLUSIONS: The results of this study suggest that the KTP laser has the potential to become a useful and reliable tool for limited lung resection. PMID- 16470527 TI - Use of a new silica clotting time for diagnosing lupus anticoagulant in patients who meet the clinical criteria for antiphospholipid syndrome. AB - The silica clotting time (SCT) is a phospholipid-dependent coagulation assay used for the laboratory diagnosis of lupus anticoagulant (LA) antibodies. The sensitivity and specificity of a new commercial SCT for identifying LA in patients who meet the clinical criteria for antiphospholipid syndrome (APS), and its association with thrombotic events were evaluated here. Forty-five patients who met the clinical criteria for APS according to the Sapporo International Consensus Statement were examined. Sixty-nine patients who did not meet the clinical criteria for APS, and 20 blood donors were used as controls. Plasma samples from the patients and controls were tested for LA using a new commercial SCT with low and high synthetic phospholipid concentrations. The results were compared with those obtained by diluted Russell's viper venom time (dRVVT) and activated partial thromboplastin time (APTT). SCT's sensitivity for identifying LA in patients who met the clinical criteria of APS was higher compared to APTT and dRVVT (53.3% vs. 31.1% and 31.1%), and the specificities of these assays were 96.6%, 100%, and 98.9%, respectively. When dRVVT was combined with SCT, and dRVVT was combined with APTT their sensitivities were 57.7% and 48.8%, and their specificities were 96.6% and 98.9%, respectively. A stepwise logistic regression analysis indicated that the combination of dRVVT with SCT was associated with total thrombotic events (odds ratio (OR)=11.5, 95% confidence interval (CI)=1.25 106.3, P=0.031) as well as with venous thrombosis (OR=4.09, 95% CI=1.16-14.43, P=0.028). According to our results, SCT is the most sensitive assay for identifying LA in patients who meet the clinical criteria for APS. Moreover, the highest sensitivity was reached with a combination of SCT and dRVVT. The method's association with total thrombotic events and venous thrombosis was in fact significant. PMID- 16470529 TI - Diagnosis of Chlamydia trachomatis infection. AB - Important progress in the diagnosis of Chlamydia trachomatis (C. trachomatis) includes the development of nucleic acid amplification techniques such as polymerase chain reaction (PCR) and ligase chain reaction (LCR). Commercial kits are available, but they are costly, sporadic in availability, must be imported, and are economically beyond the reach of common people. To overcome this limitation, most research laboratories have standardized their in-house-developed PCR methods for diagnosing this infection. However, each laboratory has to spend a great deal of time and money to accomplish this. Published reports do not always elaborate the steps involved in standardizing a test so that it can immediately be reproduced in another setting. In the present study we attempted to elaborate the steps involved in standardizing a sensitive and specific PCR technique followed by hybridization with specific C. trachomatis probe to diagnose this infection in cervical, introital, and urine specimens, and used it to determine the infection rate in a clinical population. PMID- 16470530 TI - Predictive value of the serum level of N-terminal pro-brain natriuretic peptide and high-sensitivity C-reactive protein in left ventricular remodeling after acute myocardial infarction. AB - This study was designed to measure the correlation between left ventricular (LV) remodeling 3 months after successful reperfusion therapy, and the levels of serum N-terminal pro-brain natriuretic peptide (NT-proBNP) and high-sensitivity C reactive protein (hs-CRP) at 3 days after acute myocardial infarction (AMI), and to determine the predictive levels of NT-proBNP and hs-CRP for LV remodeling. A total of 106 patients with first AMI were included in this study. Each patient was examined by echocardiography (ECG) on the third day and third month after AMI, and LV remodeling was determined by the difference in the LEV end-diastolic volume (LVEDV) between the third day and the third month. Serum NT-proBNP and hs CRP level were measured 3 days after AMI. Then the correlation between the 3 months change in LVEDV and the levels of serum NT-proBNP or hs-CRP was determined. In addition, sensitivity and specificity were calculated with a receiver operating characteristic (ROC) curve to identify correspondence with LV remodeling (defined as the change rate of LVEDV >20%). Our results showed that the correlation coefficients with the change of LVEDV were 0.706 for serum NT-pro BNP (P<0.001) and 0.596 for hs-CRP (P<0.05). With a cutoff value of 0.2, the area under the ROC curve (AUC) was 0.894 for NT-proBNP and 0.825 for hs-CRP. Although the AUC did not statistically differ between NT-proBNP and hs-CRP, NT-proBNP is more effective than serum hs-CRP as a marker to predict LV remodeling. PMID- 16470531 TI - Mutation analysis of Crouzon syndrome in Taiwanese patients. AB - Crouzon syndrome is an autosomal-dominant disorder that causes premature fusion of the cranial suture. Crouzon, Pfeiffer, and Apert syndromes are caused by mutations in the extracellular, third immunoglobulin-like domain, and adjacent linker regions (exons IIIa and IIIc) of the fibroblast growth factor receptor 2 (FGFR2) gene. We screened 12 Crouzon syndrome patients for mutations in exons IIIa and IIIc of the FGFR2 gene by polymerase chain reaction (PCR) and direct sequencing. Mutations were detected in nine of 12 patients at amino acid positions 278, 281, 289, 342, and 354. More than half of the studied Crouzon patients carried a mutation resulting in either the loss or gain of a cysteine residue. A novel missense Ser354Phe substitution at exon IIIc of the human FGFR2 gene was found. According to our results, sequencing analysis of IgIII domain of the FGFR2 gene can lead to a genetic diagnosis of Crouzon syndrome. PMID- 16470532 TI - Identification of the four most common beta-globin gene mutations in Greek beta thalassemic patients and carriers by PCR-SSCP: advantages and limitations of the method. AB - In the present study we investigated whether the single-strand conformational polymorphism (SSCP) method could be employed to identify (rather than simply detect) the four most common beta-globin gene mutations in the Greek population: IVS-I-110, Cd39, IVS-I-1, and IVS-I-6. Using DNA from 50 beta-thalassemic patients and carriers, we amplified by PCR the appropriate 238-bp region of the human beta-globin gene, analyzed the reaction products by nondenaturing polyacrylamide gel electrophoresis, and visualized the bands by silver staining. Single-stranded DNA (ssDNA) fragments showed a reproducible pattern of bands that was characteristic of the mutations present. With the use of control samples containing six of the 10 possible combinations of the four most common beta globin gene mutations, we were able to predict the mutations present in a quarter of the patients studied. Our predictions were confirmed independently by the amplification refractory mutation system (ARMS) method. We conclude that this non radioactive PCR-SSCP method can be used to reliably identify mutations in patients, provided that suitable controls are available. Moreover, the method is easy to apply to the identification of mutations in carriers, which makes it particularly useful for population screening. PMID- 16470533 TI - Haplotype association analysis for late onset diseases using nuclear family data. AB - In haplotype-based association studies for late onset diseases, one attractive design is to use available unaffected spouses as controls (Valle et al. [1998] Diab. Care 21:949-958). Given cases and spouses only, the standard expectation maximization (EM) algorithm (Dempster et al. [1977] J. R. Stat. Soc. B 39:1-38) for case-control data can be used to estimate haplotype frequencies. But often we will have offspring for at least some of the spouse pairs, and offspring genotypes provide additional information about the haplotypes of the parents. Existing methods may either ignore the offspring information, or reconstruct haplotypes for the subjects using offspring information and discard data from those whose haplotypes cannot be reconstructed with high confidence. Neither of these approaches is efficient, and the latter approach may also be biased. For case-control data with some subjects forming spouse pairs and offspring genotypes available for some spouse pairs or individuals, we propose a unified, likelihood based method of haplotype inference. The method makes use of available offspring genotype information to apportion ambiguous haplotypes for the subjects. For subjects without offspring genotype information, haplotypes are apportioned as in the standard EM algorithm for case-control data. Our method enables efficient haplotype frequency estimation using an EM algorithm and supports probabilistic haplotype reconstruction with the probability calculated based on the whole sample. We describe likelihood ratio and permutation tests to test for disease haplotype association, and describe three test statistics that are potentially useful for detecting such an association. PMID- 16470534 TI - Increased immature hematopoietic progenitor cells CD34+/CD38dim in myelodysplasia. AB - BACKGROUND: Myelodysplastic syndromes (MDS) are clonal disorders affecting hematopoietic progenitor cells (HPC). Despite the relevance of clonal CD34+ cells in developing MDS, only few studies analyze the phenotype of this cell population. The aim of this study was to evaluate phenotypic changes on HPC in MDS that could reflect abnormalities in the differentiation process of stem cells. METHODS: We analyzed the expression of CD38 and HLA-DR on CD34+ cells by flow cytometry in 36 patients with MDS, as well as in healthy donors (n = 12) and patients with other hematological disorders: non-Hodgkin lymphomas and multiple myeloma, both in complete remission (CR) (n = 32); acute lymphoblastic leukemia in CR (n = 17); de novo acute myeloblastic leukemia (AML) at diagnosis (n = 22) and in CR (n = 37); and AML secondary to MDS at diagnosis (n = 19). Cases with available karyotype were grouped according to the International Prognostic Scoring System (IPSS). RESULTS: Compared to normal BM, the fraction of immature HPC, characterized as CD34+bright, intermediate FSC/SSC, and CD38dim, was significantly increased in high risk MDS and secondary AML, but not in low risk MDS, (P < or = 0.001, P = 0.03, and P = 0.7). De novo AML showed decreased immature HPC. High numbers of immature HPC correlated with higher IPSS risk groups (P = 0.05) and showed significant impact on disease progression (P = 0.03). CONCLUSION: Our study confirms that evaluation of CD38 expression pattern on HPC is an easy and reproducible test that allows evaluating the immature subset of progenitor cells. Increased immature HPC in high risk MDS and secondary AML may reflect blocked differentiation of CD34+ cells in these diseases. PMID- 16470535 TI - Simultaneous detection of antibody binding and cytotoxicity in flow cytometry crossmatch for renal transplantation. AB - BACKGROUND: The anti-HLA antibody can be detected using either a complement dependent lymphocytotoxicity (CDC) assay or a flow cytometry crossmatch (FCXM) in renal transplantation. Discordant results are often obtained because the two methods detect different reaction phases between the donor lymphocytes and the recipient sera. This study was intended to confirm that antibody binding and cytotoxicity to the lymphocytes can be detected simultaneously in a single FCXM assay, cytotoxic FCXM. METHODS: In the cytotoxic FCXM, the antibody binding to the lymphocytes was measured using anti-IgG-FITC, and the cytotoxicity using 7 aminoactinomycin D (7-AAD) after adding complement. For an evaluation of two test parameters, the cytotoxicity test moiety (dead-cell percentage) was compared with the anti-human globulin (AHG)-CDC, and the antibody-binding test moiety (sample/control fluorescence ratio) with the conventional FCXM in 77 positive and 30 negative crossmatches. RESULTS: In the cytotoxic FCXM, both antibody binding and cytotoxicity could be assessed in a single anti-IgG-FITC/7-AAD plot. Regarding the correlation between the presence of HLA antibodies and the test result, the cytotoxicity parameter (r = 0.55) appeared to be more suitable than that of the AHG-CDC (r = 0.50) but the antibody-binding parameter (r = 0.83) was worse than that of the conventional FCXM (r = 0.93). The sensitivity of both parameters of the cytotoxic FCXM was not significantly different from each conventional counterpart (P = 0.33 and P = 0.22, respectively). CONCLUSIONS: The simultaneous detection of Ab binding and cytotoxicity was possible by the cytotoxic FCXM with the test efficiencies similar to the conventional counterparts. If this new assay is improved through the further studies to optimize the critical assay variables, this may be used as an alternative to the conventional assays to acquire more information on the characteristics of the recipient's HLA alloantibodies. PMID- 16470536 TI - Allelotyping analysis at chromosome arm 8p of high-grade prostatic intraepithelial neoplasia and incidental, latent, and clinical prostate cancers. AB - In this study, we used 7 informative microsatellite markers at 8p22, 23.1, and 23.2 in Japanese patients to compare frequency of loss of heterozygosity (LOH) in 53 lesions of high-grade prostatic intraepithelial neoplasia (HGPIN), 38 cases (38 lesions) of incidental prostate cancer (IPC), 31 cases (41 lesions) of latent prostate cancer (LPC), and 102 cases (168 lesions) of clinical prostate cancer (CPC). The frequency of LOH at 8p22-23.2 with at least 1 marker was 0%, 33%, 57%, and 51% in the HGPIN, IPC, LPC, and CPC cases, respectively. No statistically significant difference was found at 8p22-23.2 between the types of prostate cancer. However, the frequency of 8p22 deletion was significantly higher in CPC and LPC cases than in IPC cases (P = 0.0003) or lesions (P = 0.0017). The frequency of LOH at 8p22 and 8p23.1 loci in high-grade tumors was significantly higher than in low-grade tumors in both the LPCs/IPCs and CPCs (P < 0.05). Allelic loss at 8p22 was significantly more frequent in CPC than in IPC (P = 0.002) and in pT4 CPC than in earlier-stage CPC (P = 0.038). These findings suggest that deletion of 8p is an important event in both the initiation and metastasis of prostate cancer. The extremely high frequency of LOH at 8p22-23.1 in high-grade tumors suggests the existence of a novel putative tumor-suppressor gene associated with the progression of prostate cancer. These results should be useful in identifying the target gene of deletion at 8p. PMID- 16470537 TI - Alterations in the expression of antioxidant genes and the levels of transcription factor NF-kappa B in relation to diabetic embryopathy in the Cohen diabetic rat model. AB - BACKGROUND: We have previously shown that oxidative stress is important in the pathogenesis of diabetes-induced anomalies in Cohen Diabetic sensitive (CDs) rat embryos and seems to interplay with genetic factors. We investigated the role of genetic factors related to the antioxidant defense mechanism in CDs rat embryos. METHODS: We studied 11.5- and 12.5-day embryos of Cohen Diabetic resistant (CDr) and CDs rats that were fed a regular diet (RD), and hence not diabetic, compared to rats fed a high-sucrose low-copper diet (HSD) where only the CDs animals became diabetic. Embryos were monitored for growth and congenital anomalies. mRNA of catalase (CAT), glutathione peroxidase (GSHpx), CuZn-SOD (SOD-superoxide dismutase), and Mn-SOD and the extent of nuclear factor kappa B (NF-kappaB) activation were assessed. RESULTS: Embryos of CDs dams fed RD were significantly smaller and had an increased rate of NTDs compared to embryos of CDr dams fed RD. When CDs dams were fed HSD, >50% of the CDs embryos were dead and 44% of the live embryos had NTDs. Live 11.5-day old embryos of CDs dams fed RD had a statistically significant increase in CAT, CuZn-SOD, and GSHpx mRNA levels compared with the levels in the CDr embryos from dams fed RD. CDs embryos from dams fed HSD showed significant overactivation of NF-kappaB compared with CDr embryos from dams fed HSD (in which activation was decreased), without any increase in the expression of SOD, CAT, and GSHpx. CONCLUSIONS: This study demonstrates that one of the genetic differences between the CDr and CDs strains fed RD is an increased expression of genes encoding for antioxidant enzymes in the CDs but inability for upregulation in diabetes. In addition, while activation of NF-kappaB is decreased in CDr on HSD, it is increased in the CDs. These differences may play a role in the increased sensitivity of the CDs embryos to diabetic-induced teratogenicity. PMID- 16470538 TI - PDGFRalpha, PDGFRbeta and KIT expression/activation in conventional chondrosarcoma. AB - Chondrosarcomas represent 20% of all primary bone sarcomas, and many studies have attempted to unravel molecular targets for future development of new therapies. The aim of this study was to investigate the expression/activation of PDGFRalpha, PDGFRbeta and KIT receptor tyrosine kinases (RTKs) as potential therapeutic targets in conventional central primary chondrosarcomas (CCS). The expression of PDGFRalpha, PDGFRbeta and KIT RTKs was detected in 16 CCSs using immunohistochemistry (IHC), and their level of expression and activation status were analysed by immunoprecipitation and western blot experiments. PDGFRalpha, PDGFRbeta and KIT cDNAs were screened to verify the presence of activating mutations and the presence of the cognate ligands was analysed by means of RT PCR. RTK gene amplification was further studied by means of fluorescence in situ hybridization (FISH) analysis. The immunophenotyping and biochemical analyses showed that the CCSs co-expressed PDGFRalpha and PDGFRbeta, with the latter showing definitively greater protein expression and phosphorylation levels. PDGFRbeta was expressed but not activated in control healthy joint cartilage, in line with no PDGFB detection. Conversely, the KIT gene product did not seem to play a relevant role. These findings, in the absence of activating mutations or an abnormal genomic profile and the presence of PDGFA and PDGFB expression, are consistent with an autocrine/paracrine loop activation of the corresponding receptors. The CCS gene profile described here offers a rationale for the use of RTK inhibitors alone or in combination with chemotherapy, and supports further investigation of RTKs and their downstream signals. PMID- 16470539 TI - Persistence of the left superior caval vein. PMID- 16470540 TI - Identification of a high-affinity binding site for dinotefuran in the nerve cord of the American cockroach. AB - The binding of the neonicotinoid insecticide dinotefuran to insect nicotinic acetylcholine receptors (nAChRs) was examined by a centrifugation method using the nerve cord membranes of American cockroaches and [3H]dinotefuran (78 Ci mmol 1). The Kd and Bmax values of [3H]dinotefuran binding were estimated to be 13.7 nM and 14.8 fmol 40 microg-1 protein respectively by Scatchard analysis. Epibatidine, an nAChR agonist, showed a rather lower affinity to the dinotefuran binding site (IC50=991 nM) than dinotefuran (IC50=5.02 nM). Imidacloprid and nereistoxin displayed lower potencies than dinotefuran but higher potencies than epibatidine. The potencies of five dinotefuran analogues in inhibiting the specific binding of [3H]dinotefuran to nerve cord membranes were determined. A good correlation (r2=0.970) was observed between the -log IC50 values of the tested compounds and their piperonyl butoxide-synergised insecticidal activities (-log LD50 values) against German cockroaches. The results indicate that a high affinity binding site for dinotefuran is present in the nerve cord of the American cockroach and that the binding of ligands to the site leads to the manifestation of insecticidal activity. PMID- 16470541 TI - Comparative toxicity of Rosmarinus officinalis L. essential oil and blends of its major constituents against Tetranychus urticae Koch (Acari: Tetranychidae) on two different host plants. AB - Bioassays of Rosmarinus officinalis L. essential oil and blends of its major constituents were conducted using host-specific strains of the two-spotted spider mite, Tetranychus urticae Koch, on bean and tomato plants. Two constituents tested individually against a bean host strain and five constituents tested individually against a tomato host strain accounted for most of the toxicity of the natural oil. Other constituents were relatively inactive when tested individually. Toxicity of blends of selected constituents indicated a synergistic effect among the active and inactive constituents, with the presence of all constituents necessary to equal the toxicity of the natural oil. PMID- 16470542 TI - The gross anatomy of the extrathoracic course of the intercostobrachial nerve. AB - Recent reports emphasize the importance of preserving the intercostobrachial nerve (ICBN) during surgical procedures (i.e., mastectomy, axillary clearance). However, a limited number of scientific reports explore the surgical anatomy of this nerve. We dissected 100 adult human formalin-fixed cadavers (200 axillae). In all the cadavers the ICBN was present with variant contributions from intercostal nerves T1, T2, T3, and T4. The arrangements of the ICBN were typed as I through VIII. The components of Type I (45% or 90 of our specimens) included a branch to the posterior antebrachial cutaneous nerve, a branch to the anterior and lateral parts of the axilla, a branch to the medial side of the arm, and a branch to the medial antebrachial cutaneous nerve. Type II (25%) describes the ICBN arising from T2 and giving off a branch to the brachial plexus. In Type III (10%), lateral cutaneous branches of T2 and T3 fuse as a common trunk and then split immediately after exiting the intercostal space to form an ICBN. In type IV (5%), T2 and T3 join distally to form an ICBN that ends as its terminal branches. Type V (5%): T3 joins T2 from the same intercostal space proximally, with Type VI (3%) showing a very proximal branching of the sensory terminal nerves. Type VII (5%) displayed a contribution from T3 and a branch to the brachial plexus with multiple terminating branches. A contribution from T3 and T4 and a branch to the brachial plexus with multiple branches of termination comprised Type VIII (2%). PMID- 16470544 TI - Psychosocial work environment and registered absence from work: estimating the etiologic fraction. AB - BACKGROUND: Evidence is growing that an adverse psychosocial work environment increases sickness absence, but little is known on the magnitude of this problem or the impact of specific factors. METHODS: Psychological demands, decision authority, skill discretion, social support from colleagues or supervisor, predictability, and meaning of work were assessed with questionnaires at baseline and sickness absence was followed-up in employers' registers for 1,919 respondents (response rate 75.2%, 68% women, mainly low-skilled jobs) from 52 Danish workplaces during a 2-year period. Etiologic fractions (EFs) were calculated with the most favorable quartiles as reference. RESULTS: In the fully adjusted model, the following EFs were found: decision authority: 12%; social support from supervisors: 8%; psychological demands: 6%; and predictability: 5%. In total, the seven psychosocial factors explained 29% of all sick-leave days. CONCLUSIONS: The results suggest that improving the psychosocial work environment among the less favorable 75% may prevent substantial amounts of absence. PMID- 16470543 TI - Temporal change in local forces and total force all over the surface of the sea urchin egg during cytokinesis. AB - We determined the tension over the entire surface of the sea urchin eggs during cytokinesis, on the basis of the intracellular pressure and cell shape. This allowed us to determine the temporal changes in both the distribution of local forces and the total force produced in the whole cell cortex. A spike-like peak at anaphase and a broader peak at the onset of furrowing were observed in the time-course of the total force. Treatment of the eggs with cytochalasin D, blebbistatin, ML-9, or ML-7 significantly lowered the total force when they inhibited cytokinesis, suggesting that the tension results mainly from the interaction between intact actin filaments and activated myosin II. Myosin II would function as a motor, not only in the furrow region, but over a wide area of the cell surface, because the sum of the tensions outside the furrow region was larger than that inside the furrow region throughout cytokinesis. The distribution of the local force revealed that a global increase in the cortical force started well before the onset of furrowing, and that the force inside the furrow region continued to increase despite the decrease in the force outside the furrow region after the onset of furrowing. The spatial and temporal patterns of the force over the entire surface support the hypothesis that there are two separate but coordinated actomyosin activation mechanisms, one of which induces global activation of the cortex and the other of which then maintains the contractility only inside the furrow region. PMID- 16470545 TI - Risk factors for sick leave due to musculoskeletal disorders among self-employed Dutch farmers: a case-control study. AB - BACKGROUND: In Dutch agriculture, musculoskeletal disorders are a main cause of sick leave. Among self-employed insured farmers, neck, shoulder, upper extremity, and back disorders accounted for 30% of the claims for sick leave of less than 1 year This case-control study set out to identify and quantify risk factors for sick leave due to musculoskeletal disorders among self-employed Dutch farmers. METHODS: Sick leave, claimed at an insurance company from 1998 to 2001 for back (SL-BP, n = 198) or neck/shoulder/upper extremity trouble (SL-EXT, n = 89) was analyzed; the controls did not file any claim in this period (n = 816). RESULTS: Multivariate analysis showed that risk factors for SL-BP were increased age (OR = 1.06 per year, CI = 1.04-1.09), body mass index (BMI) >27 (OR = 1.93, CI = 1.2 3.2), smoking (OR = 1.90, CI = 1.2-2.9), former pain (OR = 3.28, CI = 2.1-5.1), tractor driving >1,000 hr/year (OR = 2.44, CI = 1.0-6.4), and "high work pace and workload" (OR = 1.59, CI = 1.0-2.4). SL-EXT was associated with pig (OR = 3.63, CI = 1.4-9.7), mushroom (OR = 6.14, CI = 1.4-27.2), or dairy/pig farming (OR = 4.56, 1.1-19.5), while age (OR = 1.10, CI = 1.06-1.14), smoking (OR = 1.79, CI = 1.0-3.2), and former pain (OR = 3.37, CI = 1.9-6.1) were also contributing. CONCLUSIONS: Prevention of sick leave of self-employed farmers should focus on life style (obesity, smoking), reducing older farmers' exposure to physical load, exposure to long-term tractor driving. Specific attention should be paid to animal and mushroom farmers. PMID- 16470546 TI - The costs of mercury exposure. PMID- 16470548 TI - The impact of menopause and lifestyle factors on blood and bone lead levels among female former smelter workers: the Bunker Hill Study. AB - OBJECTIVE: The Bunker Hill mine in Idaho operated from 1886 to 1981. In the 60's and 70's it provided approximately 25% of the primary lead in the United States. Women first began working on the production and maintenance lines in 1972. This study examines the impact of menopause and several occupational and lifestyle factors as determinants of blood and bone lead levels among 73 female former smelter workers. METHODS: Blood lead levels were analyzed using graphite furnace atomic absorption spectroscopy. The (109)Cd K X-ray fluorescence system was used to measure calcaneus and tibia bone lead content. Information was obtained on reproductive history, alcohol and cigarette consumption, education, and hormone replacement therapy (HRT). RESULTS: Postmenopausal women (n = 47) had significantly (P < 0.002) higher blood lead levels (3.48 microg/dl) than did premenopausal women (n = 26) (2.19 microg/dl). The best predictors of blood lead levels were smoking (>10 or < or =10 cigarettes/day), natural menopause, technical or community college education, manager or technical worker, and past or present use of HRT. The best predictors of calcaneus bone lead levels (P < 0.2) were technical workers, such as miner; natural menopause; and smoking >10 cigarettes/day; level of education; 1-2 pregnancies; and age (>60 years). CONCLUSIONS: Lead exposure results in higher blood lead levels especially during menopause. PMID- 16470549 TI - Mental retardation and prenatal methylmercury toxicity. AB - BACKGROUND: Methylmercury (MeHg) is a developmental neurotoxicant; exposure results principally from consumption of seafood contaminated by mercury (Hg). In this analysis, the burden of mental retardation (MR) associated with methylmercury exposure in the 2000 U.S. birth cohort is estimated, and the portion of this burden attributable to mercury (Hg) emissions from coal-fired power plants is identified. METHODS: The aggregate loss in cognition associated with MeHg exposure in the 2000 U.S. birth cohort was estimated using two previously published dose-response models that relate increases in cord blood Hg concentrations with decrements in IQ. MeHg exposure was assumed not to be correlated with native cognitive ability. Previously published estimates were used to estimate economic costs of MR caused by MeHg. RESULTS: Downward shifts in IQ resulting from prenatal exposure to MeHg of anthropogenic origin are associated with 1,566 excess cases of MR annually (range: 376-14,293). This represents 3.2% of MR cases in the US (range: 0.8%-29.2%). The MR costs associated with decreases in IQ in these children amount to $2.0 billion/year (range: $0.5-17.9 billion). Hg from American power plants accounts for 231 of the excess MR cases/year (range: 28-2,109), or 0.5% (range: 0.06%-4.3%) of all MR. These cases cost $289 million (range: $35 million-2.6 billion). CONCLUSIONS: Toxic injury to the fetal brain caused by Hg emitted from coal-fired power plants exacts a significant human and economic toll on American children. PMID- 16470550 TI - Finnish Institute of Occupational Health (FIOH): prevention and detection of asbestos-related diseases, 1987-2005. AB - BACKGROUND: Between 1987 and 1992, the Finnish Institute of Occupational Health (FIOH) initiated and implemented the Asbestos Program that aimed at reducing asbestos-related risks. It was a cooperative effort between government authorities, labor market organizations, and health care and labor protection personnel. METHODS: During the Program and its follow-up since 1993 several preventive actions were taken, and considerable new legislation and official instructions were issued. RESULTS: Approximately 200,000 people in Finland have been exposed to asbestos in their work. Through the cooperative efforts of government, labor, health care and worker protection programs, the import of asbestos was ceased in 1993 with a few exceptions. Almost all imports ceased in 2005 following European Union directives. Regulation of asbestos abatement companies has been greatly increased. The occupational exposure limit has been reduced from 2.0 fibers/cm(3) to the present 0.1 fibers/cm(3). Asbestos-related diseases are closely monitored and education of health care providers regarding the occupational source of these conditions is now emphasized. CONCLUSIONS: The success of the primary goal of the Program, a reduction in asbestos-related morbidity, will not be fully realized for many decades. PMID- 16470551 TI - Germline LEMD3 mutations are rare in sporadic patients with isolated melorheostosis. AB - To further explore the allelic heterogeneity within the group of LEMD3-related disorders, we have screened a larger series of patients including 5 probands with osteopoikilosis or Buschke-Ollendorff syndrome (BOS), 2 families with the co occurrence of melorheostosis and BOS, and 12 unrelated patients with isolated melorheostosis. Seven novel LEMD3 mutations were identified, all predicted to result in loss-of-function of the protein. We confirm that loss-of-function mutations in the LEMD3 gene can result in either osteopoikilosis or BOS. However, LEMD3 germline mutations were only found in two melorheostosis patients belonging to a different BOS family and one sporadic patient with melorheostosis. The additional presence of osteopoikilosis lesions in these patients seemed to distinguish them from the group of sporadic melorheostosis patients where no germline LEMD3 mutation was identified. Somatic mosaicism for a LEMD3 mutation in the latter group was also not observed, and therefore we must conclude that the genetic defect in the majority of sporadic and isolated melorheostosis remains unknown. PMID- 16470552 TI - Mutation profile of the MYO7A gene in Spanish patients with Usher syndrome type I. AB - Usher syndrome type I is the most severe form of Usher syndrome. It is an autosomal recessive disorder characterized by profound congenital sensorineural deafness, retinitis pigmentosa, and vestibular abnormalities. Mutations in the myosin VIIA gene (MYO7A) are responsible for Usher syndrome type 1B (USH1B). This gene is thought to bear greatest responsibility for USH1 and, depending on the study, has been reported to account for between 24% and 59% of USH1 cases. In this report a mutation screening of the MYO7A gene was carried out in a series of 48 unrelated USH1 families using single strand conformation polymorphism analysis (SSCP) and direct sequencing of those fragments showed an abnormal electrophoretic pattern. Twenty-five mutations were identified in 23 out of the 48 families studied (47.9%). Twelve of these mutations were novel, including five missense mutations, three premature stop codons, three frameshift, and one putative splice-site mutation. Based on our results we can conclude there is an absence of hot spot mutations in the MYO7A gene and that this gene plays a major role in Usher syndrome. PMID- 16470553 TI - Five novel androgen receptor gene mutations associated with complete androgen insensitivity syndrome. AB - Mutations in the androgen receptor (AR) gene result in androgen insensitivity syndrome (AIS). We have identified five novel mutations that result in a complete loss in AR function and are associated with complete AIS. The mutations span all three AR major functional domains. In two cases, the loss of AR function could be explained on the basis of the current knowledge of AR molecular structure and function. N-terminal mutation c.256C>T (p.Gln86X) leads to an early stop codon and abolishes all DNA and ligand binding. The DNA-binding domain mutation c.1685G>A (p.Cys562Tyr) is located in the N-terminal part of the first zinc finger; a mutation in this position is likely to impair the association of the mutated AR with the androgen response element of target genes. The splice site mutation at intron 2/exon 3 junction (c.1766-1G>A) is shown to lead to c.1765_1766 ins69 (p.[Gly589_Lys590ins23;Gly589Glu]). The two novel ligand binding domain mutations identified were recreated by site-directed mutagenesis. Both mutations c.2171G>T (p.Gly724Val) and c.2435T>C (p.Leu812Pro) abolished AR ligand binding and severely impaired AR mediated transactivation. Residue p.Gly724 is located in the ligand binding domain, between helices 3 and 4. This region is known to be involved not only in ligand binding but also in AR N/C terminal interactions. The mutation p.Leu812Pro is located in the C-terminal end of helix 8. This domain is highly conserved and critical for ligand binding. This study extends current understanding of AR mutations associated with CAIS. PMID- 16470555 TI - De novo gene conversion in the RCA gene cluster (1q32) causes mutations in complement factor H associated with atypical hemolytic uremic syndrome. AB - Many of the complement regulatory genes within the RCA cluster (1q32) have arisen through genomic duplication and the resulting high degree of sequence identity is likely to predispose to gene conversion events. The highest degree of identity is between the genes for factor H (CFH) and five factor H-related proteins--CFHL1, CFHL2, CFHL3, CFHL4, and CFHL5. CFH mutations are associated with atypical hemolytic uremic syndrome (aHUS). In the Newcastle cohort of 157 aHUS patients we have identified CFH mutations in 25 families or individuals. Eleven of these 25 independent mutations are either c.3226C>G,Q1076E; c.3572C>T,S1191L; c.3590T>C,V1197A or combined c.3572C>T,S1191L/c.3590T>C,V1197A. Sequence analysis shows that all four of these changes could have arisen as a result of gene conversion between CFH and CFHL1. Analysis of parental samples in two patients with S1191L/V1197A has shown that the changes are de novo thus providing conclusive evidence that gene conversion is the mutational mechanism in these two cases. To confirm that S1191L and V1197A are disease predisposing we examined their functional significance in three ways - analysis of the C3b/C3d binding characteristics of recombinant mutant S1191L/V1197A protein, heparin affinity chromatography and haemolytic assays of serum samples from aHUS patients carrying these changes. The results showed that these changes resulted in impaired C3b binding and a defective capacity to control complement activation on cellular surfaces. We, therefore, provide conclusive evidence that gene conversion is responsible for functionally significant CFH mutations in aHUS. PMID- 16470554 TI - Vacuolating megalencephalic leukoencephalopathy with subcortical cysts: functional studies of novel variants in MLC1. AB - Nine new unrelated patients presenting vacuolating myelinopathy with subcortical cysts were identified and analyzed for variations in the MLC1 gene. We detected 12 mutations (p.Leu37fs, p.Met80Val, p.Leu83Phe, p.Pro92Ser, p.Ser93Leu, p.Ile108fs, p.Gly130Arg, p.Cys171fs, p.Glu202Lys, p.Ser269Tyr, p.Ala275Asn, and p.Leu310_311insLeu) of which nine were novel. In one patient we did not detect mutations. Using a heterologous system, three new missense variants (p.Glu202Lys, p.Ser269Tyr, and p.Ala275Asn) and a single leucine insertion (p.Leu310insLeu)- lying in a stretch of seven leucines--were functionally assayed by determining total protein levels and mutant protein expression at the plasma membrane. No correlation was observed between mutation, clinical features, and plasma membrane expression of mutant protein. PMID- 16470557 TI - Selective immobilization of proteins onto solid supports through split-intein mediated protein trans-splicing. PMID- 16470558 TI - Total syntheses of hexacyclinol, 5-epi-hexacyclinol, and desoxohexacyclinol unveil an antimalarial prodrug motif. PMID- 16470559 TI - Measuring orbitals: provocation or reality? PMID- 16470560 TI - A supramolecular bundling approach toward the alignment of conjugated polymers. PMID- 16470561 TI - Synthesis of substituted chromenes through the DABCO-catalyzed reaction of but-3 yn-2-one and methyl propiolate with salicyl N-tosylimines (DABCO = 1,4 diazabicyclo[2.2.2]octane). AB - The DABCO-catalyzed (DABCO = 1,4-diazabicyclo[2.2.2]octane) reaction of but-3-yn 2-one and methyl propiolate with salicyl N-tosylimines yields highly functionalized chromenes and has been thoroughly investigated; the rational mechanism for the reaction has been demonstrated on the basis of 1H NMR spectroscopic investigation. PMID- 16470562 TI - Synthesis of 7-deoxypancratistatin from carbohydrates by the use of olefin metathesis. AB - The stereocontrolled synthesis of (+)-7-deoxypancratistatin is described. The convergent synthesis has been achieved by two different strategies, both of which commence from a pentose and piperonal. The latter is converted into allylic bromide 7, which is then coupled with a protected methyl 5-deoxy-5-iodo-D ribofuranoside in the presence of zinc metal. The first strategy involves a total of only 13 steps from D-ribose and piperonal, but suffers from a low yield in the zinc-mediated reaction between ribofuranoside 9, benzylamine, and bromide 7. The second strategy involves a total of 18 steps from D-xylose and piperonal. The former is converted into ribofuranoside 28, which is coupled with bromide 7 in the presence of zinc, and this is followed by ring-closing olefin metathesis. Subsequent Overman rearrangement, dihydroxylation, and deprotection then affords the natural product. PMID- 16470564 TI - Fragmentation of the deprotonated ions of peptides containing cysteine, cysteine sulfinic acid, cysteine sulfonic acid, aspartic acid, and glutamic acid. AB - We examined the fragmentation of the electrospray-produced [M-H]- and [M-2H]2- ions of a number of peptides containing two acidic amino acid residues, one being aspartic acid (Asp) or glutamic acid (Glu), and the other being cysteine sulfinic acid [C(SO2H)] or cysteine sulfonic acid [C(SO3H)], on an ion-trap mass spectrometer. We observed facile neutral losses of H2S and H2SO2 from the side chains of cysteine and C(SO2H), respectively, whereas the corresponding elimination of H2SO3 from the side chain of C(SO3H) was undetectable for most peptides that we investigated. In addition, the collisional activation of the [M H]- ions of the C(SO2H)-containing peptides resulted in the cleavage of the amide bond on the C-terminal side of the C(SO2H) residue. Moreover, collisional activation of the [M-2H]2- ions of the above Asp-containing peptides led to the cleavage of the backbone N-Calpha bond of the Asp residue to give cn and/or its complementary [zn-H2O] ions. Similar cleavage also occurred for the singly deprotonated ions of the otherwise identical peptides with a C-terminal amide functionality, but not for the [M-H]- ions of same peptides with a free C terminal carboxylic acid. Furthermore, ab initio calculation results for model cleavage reactions are consistent with the selective cleavage of the backbone N Calpha bond in the Asp residue. PMID- 16470563 TI - Umbilical cord pseudocyst in a fetus with Down syndrome. PMID- 16470565 TI - Semiquantitative analysis of isomeric oligosaccharides by negative-ion mode UV MALDI TOF postsource decay mass spectrometry and their fragmentation mechanism study at N-acetyl hexosamine moiety. AB - Postsource decay (PSD) spectra of isomeric neutral lactooligosaccharide mixtures were measured from the chlorinated molecules [M + Cl]- by negative-ion mode ultraviolet matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (UV-MALDI TOF MS) to estimate quantitatively the mixing ratios in their mixtures. The PSD ions specific to each isomeric structure were used to distinguish the linkage and branching isomers, and the molar ratios of the isomers were estimated from their ion abundances. The relative ion abundances changed linearly in the PSD spectra of the mixtures of the isomers as their molar ratio was varied in the analyte solutions. Therefore, the molar ratios of the isomers in the analyte mixtures could be estimated semiquantitatively. In addition, we studied their fragmentation mechanisms in N-acetyl hexosamines such as GlcNAc, which enabled us to quantitatively analyze the structures of the isomers of lactooligosaccharides. The conjugated systems elongate in the chemical species of the Z-type fragmentation on the 3-linked GlcNAc owing to the acetoamido groups at the C-2 positions, which made the chemical species of the Z type ions stable. The glycosyl bonds of the front of GlcNAc cleaved easily as a C type fragmentation because the negative charge at the anomeric position could be delocalized to the carbonyl oxygen atom at the acetoamido group of GlcNAc. These factors caused the stabilization of the chemical species of the C/Z fragment ions produced by the double cleavage around GlcNAc. PMID- 16470567 TI - Fragmentation study of imatinib and characterization of new imatinib metabolites by liquid chromatography-triple-quadrupole and linear ion trap mass spectrometers. AB - Imatinib (Gleevec) is an anticancer drug that inhibits specific protein kinases involved in cell proliferation. Whereas this drug is considered to have opened a new era, various mechanisms of resistance have been associated with imatinib relapse. Drug disposition in cancer cells including influx, efflux and drug metabolism is one mechanism that remains to be more thoroughly investigated. Moreover, recent genomic studies have revealed that some isozymes of cytochrome P450 (CYP) are possibly associated with the treatment outcome. Therefore, this research paper investigates the role of the activity of CYP1A1, 1A2, 1B1, 3A4, 4F2 and 4F3A/B on the fate of imatinib. First, a study of imatinib fragmentation was effected using electrospray triple-quadrupole and linear ion trap tandem mass spectrometers (MSn). Accurate mass determinations were performed at enhanced mass resolution for the identification of some product ions that were not predicted by two fragmentation softwares. Whereas the quadrupole MS was not designed for accurate mass measurement, delta mass errors were below 20 ppm. Then, a biotransformation study was effected in vitro. Imatinib metabolites were produced in microsomal incubations containing CYP isozymes. Imatinib and metabolites were extracted from incubation mixtures by protein precipitation, and supernatants were injected into a liquid chromatography equipment coupled with MS(n). Hydrophobic interaction liquid chromatography resolved one demethylated-, two hydroxy- and three N-oxide metabolites. Various rates of metabolite formation were observed between CYP isozymes. Liquid chromatography with deuterium oxide containing mobile phase (H/D exchange) or incorporation of (18)O from H(2) (18)O added in the incubations was performed to elucidate the metabolite structure. Various MS(n) product scans (n < or = 4) were acquired on the linear ion trap or on the triple-quadrupole MS. Postulated structures of new metabolites are addressed. PMID- 16470568 TI - Designer drugs 2,5-dimethoxy-4-bromo-amphetamine (DOB) and 2,5-dimethoxy-4-bromo methamphetamine (MDOB): studies on their metabolism and toxicological detection in rat urine using gas chromatographic/mass spectrometric techniques. AB - Studies are described on the metabolism and the toxicological analysis of the amphetamine-derived designer drug 2,5-dimethoxy-4-bromo-amphetamine (DOB) and its corresponding N-methyl analogue 2,5-dimethoxy-4-bromo-methamphetamine (MDOB) in rat urine using gas chromatographic/mass spectrometric techniques. The identified metabolites indicated that DOB was metabolized by O-demethylation followed by oxidative deamination to the corresponding ketone as well as deamination followed by reduction to the corresponding alcohol. Other metabolic pathways were O,O bisdemethylation or hydroxylation of the side chain followed by O-demethylation and deamination to the corresponding alcohol. The expected oxo compound after deamination could not be detected. All metabolites carrying hydroxy groups were found to be partly excreted in the conjugated form. MDOB underwent O demethylation, O,O-bisdemethylation, or hydroxylation of the side chain followed by O-demethylation. Additional N-demethylation to DOB occurred, including the above-mentioned metabolites. Again, all metabolites carrying hydroxy groups were found to be partly excreted in the conjugated form. The authors' systematic toxicological analysis (STA) procedure using full-scan GC/MS after acid hydrolysis, liquid-liquid extraction, and microwave-assisted acetylation allowed the detection of an intake of a dose of DOB and MDOB in rat urine that corresponds to a common drug user's dose. Assuming a similar metabolism, the described STA procedure in human urine should be suitable as proof of an intake of DOB and MDOB. PMID- 16470569 TI - Capillary liquid chromatography/atmospheric-pressure matrix-assisted laser desorption/ionisation ion trap mass spectrometry: a comparison with liquid chromatography/matrix-assisted laser desorption/ionisation time-of-flight and liquid chromatography/electrospray ionisation quadrupole time-of-flight for the identification of tryptic peptides. AB - The atmospheric-pressure matrix-assisted laser desorption/ionisation quadrupole ion trap (AP-MALDI-QIT) analysis of tryptic peptides is reported following capillary liquid chromatographic (LC) separation and direct analysis of a protein digest. Peptide fragments were identified by peptide mass fingerprinting from mass spectrometric data and sequence analysis obtained by tandem mass spectrometry of the principal mass spectral peaks using a data-dependent scanning protocol. These data were compared with those from mass spectrometric analysis using capillary LC/MALDI-time-of-flight (TOF) and capillary LC/electrospray ionisation (ESI)-quadrupole TOF. For all three configurations the resulting data were searched against the MSDB database, using MASCOT and the sequence coverage compared for each technique. Complementary data were obtained using the three techniques. PMID- 16470570 TI - Neonatal outcome of a prenatally detected 46,XX/46,XY true hermaphrodite. PMID- 16470571 TI - A comparison of the electron ionization and electrospray behaviour of some N,N' disubstituted hexahydropyrimidines. AB - The behaviour of some N,N'-disubstituted hexahydropyrimidines, a class of naturally occurring compounds of biological and biomedical interest, has been studied in both electron ionization (EI) and electrospray ionization (ESI) modes coupled with collisional experiments (ESI-MSn). In both techniques, the [M-H]+ ions are highly abundant, even if their formation is generated by two different mechanisms, i.e. H. loss from the M+. species in the case of EI and hydride (H-) abstraction from the molecules in the case of ESI. Furthermore, due to the low, step-by-step internal energy deposition typical of collisional experiments performed in an ion trap mass spectrometer, different fragment ions were observed in EI and ESI-MSn collisions. In both cases, the ions can be related to the original structure and allow us to identify the positions in which the different substituents are present. PMID- 16470572 TI - Fetal paroxysmal supraventricular tachycardia without heart failure leading to ischemic damage. PMID- 16470573 TI - Favourable outcome in a pregnancy with concomitant maternal and fetal osteogenesis imperfecta associated with a novel COL1A2 mutation. PMID- 16470574 TI - Bilateral fetal hyperechogenic kidneys associated with normal amniotic fluid: an ethical dilemma in a normal variant? PMID- 16470575 TI - Amniocentesis for karyotyping prior to induction of abortion at second trimester. PMID- 16470576 TI - Structural identification of isomers present in technical perfluorooctane sulfonate by tandem mass spectrometry. AB - The structures of isomeric by-products of technical perfluorooctane sulfonate (PFOS, C8F17SO3-) were characterized by high-performance liquid chromatography coupled to mass spectrometry. Ion trap (IT) as well as triple quadrupole (TQ) tandem mass spectrometry (MS/MS) identified ten PFOS isomers with best results being achieved by ion trap MS/MS. In addition to the linear isomer, the main by products were isomers with geminal diperfluoromethyl groups and perfluoromethyl substituted compounds. Purified fractions of the perfluoromethyl isomers allowed the elucidation of structures and the assignment of the retention times of perfluoroisopropyl-, 5-perfluoromethyl-, 4-perfluoromethyl-, 3-perfluoromethyl- and 1-perfluoromethyl-PFOS. The concentration of 2-perfluoromethyl-PFOS was too low in technical PFOS for a proper identification. Retention times of the geminal substituted diperfluoromethyl isomers could not be properly assigned due to co elutions. The number and position of the CF3 groups can be derived from the MS/MS spectra, which allow an unequivocal identification of the isomers. TQ-MS/MS also allowed us to identify most of the isomers. PMID- 16470577 TI - Prenatal counselling of small bowel atresia: watch the fluid! AB - OBJECTIVE: To evaluate polyhydramnios as a sign of extreme disproportion of atretic segments in small bowel atresia (SBA). METHODS: Twenty-eight patients with a prenatal diagnosis (PD) of SBA undergoing neonatal surgical treatment were reviewed retrospectively. Parameters recorded were gestational age, birth weight, surgical procedure, rate of complications, parenteral nutrition (PN) days and length of stay (LOS). Patients were divided into two groups: Group A with delayed anastomosis and Group B with direct anastomosis. RESULTS: Seventeen subjects were in Group A while 11 were in Group B. The two groups did not differ with regard to gestational age at diagnosis, birth weight and obstetrical management. Polyhydramnios was present in both Group A (64.7%) and Group B (9%) (p < 0.05). Patients in Group A needed a longer period on PN, had a longer LOS and exhibited significantly higher rates of complication. CONCLUSION: In the absence of other malformations, association of dilated bowel loops and polyhydramnios is highly predictive of severe SBA, which can in no instance be amenable to one-stage reconstruction. Because of its consequences on postnatal treatment, such information should be conveyed to the prospective parents at the time of counselling. PMID- 16470578 TI - Stability studies of testosterone and epitestosterone glucuronides in urine. AB - The stability of testosterone glucuronide (TG), epitestosterone glucuronide (EG) and the T/E ratio in urine has been studied. Samples were analyzed by gas chromatography coupled to mass spectrometry (GC/MS). Urine samples were submitted to a solid-liquid cleanup followed by extraction of unconjugated testosterone (T) and epitestosterone (E) with tert-butyl methyl ether (free fraction). The remaining aqueous phase was hydrolyzed with beta-glucuronidase and extracted at alkaline pH with n-pentane. Analytes were analyzed by GC/MS as their enol trimethylsilyl (TMS) derivatives. The urine for stability testing was obtained from an excretion study after the administration of T to healthy volunteers. The homogeneity of the sample was verified before starting the stability study. The stability of TG and EG was evaluated at different storage conditions. For long term stability testing, analyte concentration in urine stored at 4 degrees C and 20 degrees C was determined at different time intervals for 22 months. For short term stability testing, analyte concentration was evaluated in urine stored at 37 degrees C for 3 and 7 days. The effect of repeated freezing (at -20 degrees C) and thawing (at room temperature) was studied for up to three cycles. Data obtained in this work demonstrated the stability of TG, EG and the T/E ratio in sterilized urine samples stored at 4 and -20 degrees C for 22 months and after going through repeated freeze/thaw cycles. Decreases in concentration were observed after 7 days of storage at 37 degrees C due to the partial cleavage of the glucuronide conjugates; however, the T/E ratio was not affected. These results show the feasibility of preparing reference materials containing TG and EG to be used for quality control purposes. PMID- 16470579 TI - Solid-phase microextraction for gas chromatographic/mass spectrometric analysis of dimethoate in human biological samples. AB - A new, simple and rapid procedure for the determination of dimethoate in urine and blood samples was developed using direct immersion solid-phase microextraction and gas chromatography/mass spectrometry. This technique required only 0.1 mL of sample, and ethion was used as internal standard. Two types of coated fibre were compared (100 microm polydimethylsiloxane, and 65 microm Carbowax/divinylbenzene). Other parameters, such as extraction temperature, adsorption and desorption time, salt addition, agitation and pH, were optimized to enhance the sensitivity of the method. Limits of detection (LODs) and quantitation (LOQs) were 50 and 100 ng/mL for urine and 200 and 500 ng/mL for blood, respectively. The method was found to be linear between the LOQ and 40 microg/mL for urine, and between the LOQ and 50 microg/mL for blood, with correlation coefficients ranging from 0.9923-0.9996. Precision (intra- and interday) and accuracy were in conformity with the criteria normally accepted in bioanalytical method validation. The mean absolute recoveries of dimethoate were 1.24 and 0.50% for urine and blood, respectively. Because of its simplicity and the fact that small volumes of sample are used, the described method can be successfully used in the diagnosis of poisoning by this pesticide, namely in those situations where the sample volume is limited, as frequently occurs in forensic toxicology. PMID- 16470580 TI - Antibody-based surface plasmon resonance detection of intact viral pathogen. AB - Surface plasmon resonance (SPR) technique was used to directly detect an intact form of insect pathogen: the baculovirus, Autographa californica multiple nuclear polyhedrosis virus (AcMNPV). An SPR sensor chip with three bio-functional layers was used to detect the intact AcMNPV: amine-reactive crosslinker with a disulfide bond that chemisorbs to gold film, Protein A, and a mouse IgG monoclonal antibody raised against a surface protein of the target viral pathogen. A two-channel (reference & test) micro-fluidic SPR system is used for reliable measurement. Bio specific response to the AcMNPV is compared with the response for tobacco mosaic virus (TMV) as control. Successive exposure of the sensor chip to both viruses verifies a specific response to AcMNPV. This serves as a prerequisite to the development of a new type of viral pathogen detection sensors. PMID- 16470581 TI - Secondary pseudotumor cerebri in pediatric oncology and hematology: an unpredictable condition of varying etiology. AB - Secondary pseudotumor cerebri (PTC) in pediatric oncology and hematology is an unpredictable and under-recognized condition. It is characterized by increased cerebrospinal fluid (CSF) pressure of more than 25 cm H(2)O in the absence of underlying structural causes. Numerous conditions and medications have been implicated in the etiology of secondary PTC. Patients may suffer from incapacitating headaches and permanent visual loss, if they are not managed properly. We retrospectively analyzed the records of eight children, diagnosed with secondary PTC with an underlying hematological or oncological disease. Our study highlights the clinical features, contributing factors, and treatment options of this syndrome in children. PMID- 16470582 TI - A second leiomyosarcoma in the urinary bladder of a child with a history of retinoblastoma 12 years following partial cystectomy. AB - This case describes a twin with bilateral retinoblastoma who developed leiomyosarcoma of the bladder at age 17 and again at 39. At 17-years of age she was diagnosed with a leiomyosarcoma of the bladder after presenting with recurrent urinary tract infections, hematuria, and dysuria. She was treated with partial cystectomy. After a 12-year disease-free interval, she was diagnosed with a second leiomyosarcoma of the bladder. This case supports the relationship between the genetic form of retinoblastoma and leiomyosarcoma and illustrates the necessity for extensive follow-up and well-defined treatment of secondary neoplasms. PMID- 16470583 TI - Molecular modeling and inhibitory activity of cowpea cystatin against bean bruchid pests. AB - Plant cystatins show great potential as tools to genetically engineer resistance of crop plants against pests. Two important potential targets are the bean weevils Acanthoscelides obtectus and Zabrotes subfasciatus, which display major activities of digestive cysteine proteinases in midguts. In this study a cowpea cystatin, a cysteine proteinase inhibitor found in cowpea (Vigna unguiculata) seeds, was expressed in Escherichia coli and purified with a Ni-NTA agarose column. It strongly inhibited papain and proteinases from midguts of both A. obtectus and Z. subfasciatus bruchids, as seen by in vitro assays. When the protein was incorporated into artificial seeds at concentrations as low as 0.025%, and seeds were consumed by the bruchids larva, dramatic reductions in larval weight, and increases in insect mortality were observed. Molecular modeling studies of cowpea cystatin in complex with papain revealed that five N terminal residues responsible for a large proportion of the hydrophobic interactions involved in the stabilization of the enzyme-inhibitor complex are absent in the partial N-terminal amino acid sequencing of soybean cystatin. We suggest that this structural difference could be the reason for the much higher effectiveness of cowpea cystatin when compared to that previously tested phytocystatin. The application of this knowledge in plant protein mutation programs aiming at enhancement of plant defenses to pests is discussed. PMID- 16470584 TI - RIP death domain structural interactions implicated in TNF-mediated proliferation and survival. AB - Death domain (DD)-containing proteins are involved in both apoptosis and survival/proliferation signaling induced by activated death receptors. Here, a phylogenetic and structural analysis was performed to highlight differences in DD domains and their key regulatory interaction sites. The phylogenetic analysis shows that receptor DDs are more conserved than DDs in adaptors. Adaptor DDs can be subdivided into those that activate or inhibit apoptosis. Modeling of six homotypic DD interactions involved in the TNF signaling pathway implicates that the DD of RIP (Receptor interacting protein kinase 1) is capable of interacting with the DD of TRADD (TNFR1-associated death domain protein) in two different, exclusive ways: one that subsequently recruits CRADD (apoptosis/inflammation) and another that recruits NFkappaB (survival/proliferation). PMID- 16470585 TI - Terminal ion pairs stabilize the second beta-hairpin of the B1 domain of protein G. AB - The effects of terminal ion pairs on the stability of a beta-hairpin peptide corresponding to the C-terminal residues of the B1 domain of protein G were determined using thermal unfolding as monitored by nuclear magnetic resonance and circular dichroism spectroscopy. Molecular dynamics (MD) simulations were also performed to examine the effect of ion pairs on the structures. Eight peptides were studied including the wild type (G41) and the N-terminal modified sequences that had the first residue deleted (E42), replaced with a Lys (K41), or extended by an additional Gly (G40). Acetylated variants were made to examine the effect of removing the positive N-terminal charge on beta-hairpin stability. The rank in stability determined experimentally is K41 > E42 approximately G41 approximately G40 > Ac-K41 > Ac-E42 approximately Ac-G41 > Ac-G40. The Tm of the K41 peptide is 12 degrees C higher than G41, while the Tm values for the acetylated peptides are less than their unacetylated forms by more than 15 degrees C. NOE cross-peaks between side-chain methylene groups at the N- and C-termini and larger CalphaH shifts compared to random values are seen for K41. The addition of 20% methanol increases the stability in K41 and G41. The MD studies complement these results by showing that the charged N-terminus is important to stability. The type of ion pair observed varies with peptide, and when formed the simulations show that the ion pair can prevent fraying of the beta-strands through electrostatic and hydrophobic contacts. Therefore, introducing favorable electrostatic interactions at the N- and C-termini can substantially enhance beta-hairpin stability and help define the structure. PMID- 16470586 TI - Characterization of linker histone H1FOO during bovine in vitro embryo development. AB - Linker histones H1 are involved in various mechanisms, such as chromatin organization and gene transcription. In different organisms, a unique subtype can be found in the oocyte, however its function remains unclear. To assess the potential involvement of this oocyte linker histone (H1FOO) in chromatin modulation, we have cloned and sequenced the bovine H1FOO cDNA and followed its mRNA profile by quantitative RT-PCR in the oocyte and throughout bovine early embryo development. The highest level of mRNA was found in the germinal vesicle (GV) oocyte and diminished constantly throughout embryo development. In the 16 cell embryo and blastocyst, respectively, the mRNA levels were 200 and 2,000 times lower than in the GV oocyte. A specific antibody raised against bovine H1FOO was used to establish protein distribution in the oocyte and preimplantation embryo by immunocytochemistry. In the GV and metaphase II (MII) oocyte, as well as in the 1-, 2- and 4-cell embryo, H1FOO was localized in the cytoplasm and nucleus. The protein was uniformly spread within the cytoplasm, while it was concentrated onto the chromatin in the nucleus. In the 8- to 16-cell embryo, H1FOO's presence diminished in the cytoplasm, although it was still strongly expressed in nucleus. In the morula and blastocyst stages, the protein was totally lacking. By its position on chromatin, H1FOO could not only be involved in chromatin conformation but could also participate in activation or repression of genes during oogenesis and embryo development before embryonic genome activation. PMID- 16470587 TI - Genetic alterations in urothelial bladder carcinoma: an updated review. AB - New oncogenes and tumor suppressor genes that play an important role in the pathogenesis of urothelial bladder carcinoma have been discovered. The objectives of this review were to summarize the most important oncogenes and tumor suppressor genes involved in urothelial carcinoma and to address their role in pathogenesis, their prognostic value, and their potential use as therapeutic targets. The collected data led the authors to propose a common pathway in which the fibroblastic growth factor receptor 3 (FGFR3) mutation seems to be the earliest genetic abnormality responsible for the transformation from normal tissue to atypia and dysplasia. Three different progression pathways were proposed: The first operative pathway is from dysplasia to superficial papillary pathologic Ta (pTa) tumors to pT1 tumors and, ultimately, to pT2 tumors with FGFR3 and tuberous sclerosis complex 1 (TSC1) the responsible genes. The second major operative pathway is from dysplasia, to carcinoma in situ, and to solid pT1 and pT2 tumors. The third pathway of progression is from dysplasia to papillary T1 and pT2 tumors. The genes involved in the last 2 pathways are the p53, serine threonine protein kinase 15 (STK15), triple-function domain (TRIO), fragile histidine triad (FHIT), p63 genes; and alterations of 20q and 5p, alterations of adhesions, angiogenesis, and matrix-remodeling gene products also are involved. Finally, murine leukemia viral oncogene homologue 1 (RAF1) and CD9 are involved in the progression from papillary pT1 tumors to pT2 tumors. PMID- 16470588 TI - Recessive dystrophic epidermolysis bullosa caused by COL7A1 hemizygosity and a missense mutation with complex effects on splicing. AB - Loss-of-function mutations in the gene encoding type VII collagen, COL7A1, are the molecular basis of the blistering skin disorder, recessive dystrophic epidermolysis bullosa (RDEB). COL7A1 maps to a region of the short arm of chromosome 3 that has been found to be deleted in many types of malignancies. We have characterized the first case of a large genomic deletion in chromosome 3p21.31 that removes COL7A1 entirely in an RDEB patient. This interstitial deletion spans 255 to 520 kb and encompasses 9 to 15 genes, but seems to have no pathological consequences other than RDEB. We show that the second, hemizygous allele of COL7A1 in this patient bears a base substitution within exon 94, c.7245G>A. This translates into an amino acid substitution, p.M2415I, and leads to a complex splicing abnormality that allows marginal levels of functional mRNA and protein to be synthesized. We propose that the leakiness of the splicing defect enables the partial rescue of collagen VII deficiency. This is consistent with the diagnosis of the moderately severe form of RDEB in the proband, at variance with the most severe form, RDEB Hallopeau-Siemens, that would arise from complete collagen VII deficiency. PMID- 16470589 TI - Mutation study of Spanish patients with hereditary hemorrhagic telangiectasia and expression analysis of Endoglin and ALK1. AB - Hereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant and age dependent vascular disorder originated by mutations in Endoglin (ENG) or activin receptor-like kinase-1 (ALK1, ACVRL1) genes. The first large series HHT analysis in Spanish population has identified mutations in 17 unrelated families. Ten different mutations in ALK1 and six in ENG genes were found. Six unrelated families had a mutation in ENG gene, four representing new mutations, p.Y258fs, pV323fs, p.F279fs (c.834_837del CTTC), and p.F279fsdupC. Eleven unrelated families harboured mutations in ALK1; ten were new mutations identified as p.H328P, p.R145fs, p.G68C, p.A377T, p.H297R, p.M376T, p.C36Y, p.H328P, p.T82del and p.R47P. Overall, ALK1 mutations (HHT2) were predominant over ENG mutations (HHT1), in agreement with data reported for other Mediterranean countries (France, Italy), but at variance with Northern Europe or North America. Endoglin expression in HHT1 or HHT2 activated monocytes and blood outgrowth endothelial cells (BOECs) from older patients was well below the theoretical 50% level expected from the HHT1 haploinsufficiency model, suggesting that the pathogenic endoglin haploinsufficiency leading to the HHT phenotype is age-dependent. Interestingly, ALK1 protein levels of HHT BOECs in some missense ALK1 mutants were similar to controls. In vitro expression of these ALK1 constructs suggests that, in addition to the haploinsufficiency model, certain ALK1 mutants may inhibit the function of the wild type allele. PMID- 16470590 TI - Functional analysis of splicing mutations and of an exon 2 polymorphic variant of SERPING1/C1NH. AB - Several sequence changes have been reported in hereditary angioedema patients in intron 2 of the SERPING1/C1NH gene, but their consequences on splicing have not been determined. We examined in cell transfection assays the consequences at the mRNA level of splicing mutations affecting either the +3 or the +5 donor site positions, or the conserved canonical splicing signals of exon 2, using mutant C1 inhibitor minigene constructs. Both intron 2 mutations, c.51+3A>G and c.51+5G>A, resulted in marked exon 2 skipping in these assays, but also yielded a large fraction of normal transcripts. We show that the c.51+3A>G mutation cosegregates with low C1 inhibitor protein levels in one family. Moreover, the second base of exon 2 of the SERPING1/C1NH gene is the site of a polymorphic variant, which has been proposed as a modifier of disease severity. We found that the c.-21C allele at this position yields low but significant levels of exon 2 skipping in transfected Hep G2 or Hep 3B cells, suggesting that this allele may contribute, at the RNA level, to more severe forms of angioedema. Furthermore, we describe a previously not detected alternative splicing of exon 3, found in peripheral blood cell mRNA but not in the liver or in hepatoma cell lines and we show that, in cultured monocytes of a patient carrying the c.51+3A>G mutation, this alternative splicing is shifted from exon 3 exclusion to skipping of both exons 2 and 3. The latter finding suggests that mutations affecting splicing of exon 2 of the SERPING1/C1NH gene may have different consequences in monocytes versus other cell types. PMID- 16470591 TI - MPL mutations in 23 patients suffering from congenital amegakaryocytic thrombocytopenia: the type of mutation predicts the course of the disease. AB - Congenital amegakaryocytic thrombocytopenia (CAMT) is a rare inherited bone marrow failure syndrome. Mutations in the gene for the thrombopoietin receptor MPL were defined as the molecular cause in CAMT patients. Extending our sequence analyses from eight to a total of now 23 CAMT patients we could verify our hypothesis of genotype-phenotype correlation in CAMT. Seven different mutations predicted to lead to a complete loss of function of the thrombopoietin receptor were found in 13 patients belonging to group CAMT I with persistently low platelet counts and a fast progression into pancytopenia. Nine different missense mutations were detected in 10 patients of group CAMT II, characterized by a transient increase in platelet counts over 50 nl(-1) during the first years of life. Using in vitro assays with hematopoietic progenitors from patients of both patient groups we could provide experimental evidence for a residual activity of the thrombopoietin receptor in CAMT II patients. PMID- 16470592 TI - Improved neuronal tract tracing using manganese enhanced magnetic resonance imaging with fast T(1) mapping. AB - There has been growing interest in using manganese-enhanced MRI (MEMRI) to detect neuronal activation, neural architecture, and neuronal connections. Usually Mn(2+) produces a very wide range of T(1) change. In particular, in neuronal tract tracing experiments the site of Mn(2+) injection can have very short T(1) while distant regions have small T(1) reductions, primarily due to dilution of Mn(2+). Most MEMRI studies use T(1)-weighted sequences, which can only give optimal contrast for a narrow range of T(1) changes. To improve sensitivity to the full extent of Mn(2+) concentrations and to optimize detection of low concentrations of Mn(2+), a fast T(1) mapping sequence based on the Look and Locker technique was implemented. Phantom studies demonstrated less than 6.5% error in T(1) compared to more conventional T(1) measurements. Using center-out segmented EPI, whole-brain 3D T(1) maps with 200-microm isotropic resolution were obtained in 2 h from rat brain. Mn(2+) transport from the rat olfactory bulb through appropriate brain structures could be detected to the amygdala in individual animals. The method reliably detected less than 7% reductions in T(1). With this quantitative imaging it should be possible to study more extensive pathways using MEMRI and decrease the dose of Mn(2+) used. PMID- 16470593 TI - Passive staining: a novel ex vivo MRI protocol to detect amyloid deposits in mouse models of Alzheimer's disease. AB - Amyloid plaques are one of the hallmarks of Alzheimer's disease (AD). This study evaluated a novel microMRI strategy based on "passive staining" of brain samples by gadoteric acid. The protocol was tested at 4.7 T on control animals and APP/PS1 mice modeling AD lesions. T(1) was strongly decreased in passively stained brains. On high-resolution 3D gradient echo images, the contrast between the cortex and subcortical structures was highly improved due to a T2* effect. The brains of APP/PS1 mice revealed plaques as hypo-intense spots. They appeared larger in long compared to short TE images. This suggests that, after passive staining, plaques caused a susceptibility effect. This easily performed protocol is a complementary method to classic histology to detect the 3D location of plaques. It may also be used for the validation of in vivo MRI protocols for plaque detection by facilitating registration with histology via post mortem MRI. PMID- 16470594 TI - Automated in vivo segmentation of carotid plaque MRI with Morphology-Enhanced probability maps. AB - MRI is a promising noninvasive technique for characterizing atherosclerotic plaque composition in vivo, with an end-goal of assessing plaque vulnerability. Because of limitations arising from acquisition time, achievable resolution, contrast-to-noise ratio, patient motion, and the effects of blood flow, automatically identifying plaque composition remains a challenging task in vivo. In this article, a segmentation method using maximum a posteriori probability Bayesian theory is presented that divides axial, multi-contrast-weighted images into regions of necrotic core, calcification, loose matrix, and fibrous tissue. Key advantages of the method are that it utilizes morphologic information, such as local wall thickness, and coupled active contours to limit the impact from noise and artifacts associated with in vivo imaging. In experiments involving 142 sets of multi-contrast images from 26 subjects undergoing carotid endarterectomy, segmented areas of each of these tissues per slice agreed with histologically confirmed areas with correlations (R(2)) of 0.78, 0.83, 0.41, and 0.82, respectively. In comparison, manually identifying areas blinded to histology yielded correlations of 0.71, 0.76, 0.33, and 0.78, respectively. These results show that in vivo automatic segmentation of carotid MRI is feasible and comparable to or possibly more accurate than manual review for quantifying plaque composition. PMID- 16470595 TI - High prevalence of CBS p.T191M mutation in homocystinuric patients from Colombia. AB - Homocystinuria is an autosomal recessive disease most commonly caused by mutations in cystathionine beta-synthase (CBS). In this study we present the mutation analysis of 36 Colombian individuals from 10 unrelated kindred, with 11 individuals clinically classified as homocystinuric. Mutation analysis of the CBS gene revealed p.T191M, a prevalent mutation in Spain and Portugal, in the homozygous state in seven of the unrelated patients. Genotype-phenotype assessment of the p.T191M homozygous patients showed a high level of variability, including different severity in one pair of affected siblings. None of the patients responded biochemically to treatment with pharmacological doses of pyridoxine and folic acid as revealed by essentially unchanged homocysteine levels. This study offered a unique opportunity to study 18 heterozygous (p.T191M/wt) relatives of the homocystinuric patients. One atypical finding was that many of them presented with above average total homocysteine levels, putting them at an increased risk for vascular disease. Cryptorchidism was present in three of the cases, one of which presented also with Klinefelter syndrome. In addition to the previously described p.T191M mutation, a new mutation, p.A288T, was identified in a single individual. In this paper we present the first characterization, at a molecular level, of patients with homocystinuria from Colombia. PMID- 16470596 TI - Tissue specificity of nonlinear dynamics in baseline fMRI. AB - In this work, recent advances in the field of nonlinear dynamics (NLD) were applied to fMRI data to examine the spatio-temporal properties of BOLD resting state fluctuations. Five human subjects were imaged during resting state (visual fixation) at 3T using single-shot gradient echo planar imaging (EPI). Respiration and cardiac signals were concurrently recorded for retrospectively removing fluctuations due to these physiologic activities. Patterns of singularity in the complex plane (PSC) and Lempel-Ziv complexity (LZ) were used to study the deterministic nonlinearity in resting state fMRI data. The results show that there is greater nonlinearity (higher PSC) and determinism (lower LZ) in gray matter compared to white matter and CSF. In addition, the removal of respiratory and cardiac pulsations decreases the nonlinearity and determinism but does not alter the relative difference between gray matter and white matter. Therefore, our results demonstrate that determinism and nonlinearity in the fMRI data are tissue-specific, suggesting that they reflect native physiologic and metabolic fluctuations and are not a result of physiologic artifacts due to respiration and cardiac pulsation. PMID- 16470597 TI - Bunched phase encoding (BPE): a new fast data acquisition method in MRI. AB - A new fast data acquisition method, "Bunched Phase Encoding" (BPE), is presented. In conventional rectilinear data acquisition, only a readout gradient (and no phase encoding gradient) is applied when k-space data are acquired. Reduction of the number of phase encoding lines by increasing the phase encoding step size often leads to aliasing artifacts. Papoulis's generalized sampling theory asserts that in some cases aliasing artifact-free signals can be reconstructed even if the Nyquist criterion is violated in some regions of the Fourier domain. In this study, Papoulis's theoretical construct is exploited to reduce the number of acquired phase encoding lines. To achieve this, k-space data are sampled along a "zigzag" trajectory during each readout; samples are acquired at a sampling frequency higher than that of the normal rectilinear acquisition. The total number of TR cycles and, hence, the total scan time can be reduced. The resultant signal-to-noise ratio (SNR) often varies across the reconstructed image when using the BPE technique, and the image SNR depends on the reconstruction method. This work is comparable to a gradient based version of parallel imaging. Evidence suggests it may serve as the basis for new opportunities for fast data acquisition in MRI. PMID- 16470600 TI - Spondyloenchondrodysplasia with spasticity, cerebral calcifications, and immune dysregulation: clinical and radiographic delineation of a pleiotropic disorder. AB - Enchondromas are a feature of several constitutional disorders of bone, and the classification of different nosologic entities is still provisional. Among these disorders, spondyloenchondrodysplasia (SPENCD), as outlined by Schorr et al. [1976], is defined by the presence of radiolucent spondylar and metaphyseal lesions that represent persistence of islands of chondroid tissue within bone. Careful review of radiographic findings is needed to distinguish SPENCD from the many other disorders combining enchondromas with spinal lesions. Even when strict criteria are applied, it appears that SPENCD is clinically heterogeneous, as some SPENCD patients are neurologically intact while others present with spasticity, mental retardation, and cerebral calcifications in different combinations, and it has been suggested that SPENCD should be divided in two types. We herein report ten individuals from six families with SPENCD and illustrate the radiographic changes. Seven individuals had CNS manifestations including spasticity, developmental delay, and late-onset cerebral calcifications. We also noted that six individuals had clinical manifestations of autoimmunity (auto-immune thrombocytopenic purpura, auto-immune hemolytic anemia, auto-immune thyroiditis, and SLE) and one had been diagnosed with immune deficiency. Neurological and autoimmune manifestations were seen in different combinations within one single family. These observations suggest that SPENCD may be a single entity defined by specific radiographic features, but with remarkably pleiotropic manifestations that include CNS disease (spasticity, mental retardation, and calcifications), as well as immune dysregulation ranging from autoimmunity to immunodeficiency. The notion of recessive inheritance hitherto assumed is challenged by the observation of two apparently dominant pedigrees. PMID- 16470599 TI - Systematic search for single nucleotide polymorphisms in a lymphoid tyrosine phosphatase gene (PTPN22): association between a promoter polymorphism and type 1 diabetes in Asian populations. AB - The protein tyrosine phosphatase, nonreceptor 22 gene (PTPN22) maps to human chromosome 1p13.3-p13.1 and encodes an important negative regulator of T-cell activation, lymphoid-specific phosphatase (Lyp). Recently, the minor allele of a single-nucleotide polymorphism (SNP) at nucleotide position 1858 (rs2476601, +1858C > T) was found to be associated with type 1 diabetes. However, the degree of the association is variable among ethnic populations, suggesting the presence of other disease-associated variants in PTPN22. To examine this possibility, we carried out a systemic search for PTPN22 using direct sequencing of PCR-amplified products in the Japanese population. Association and linkage studies were also conducted in 1,690 Japanese samples, 180 Korean samples, and 472 Caucasian samples from 95 nuclear families. We identified five novel SNPs, but not the +1858C > T SNP. Of these two frequent SNPs, -1123G > C, and +2740C > T were in strong linkage disequilibrium (LD), and the -1123G > C promoter SNP was associated with acute-onset but not slow-onset type 1 diabetes in the Japanese population (odds ratio [OR] = 1.42, 95% CI = 1.07-1.89, P = 0.015). This association was observed also in Korean patients with type 1 diabetes (Mantel Haenszel chi2= 6.543, P = 0.0105, combined OR = 1.41 95% CI = 1.09-1.82). Furthermore, the affected family-based control (AFBAC) association test and the transmission disequilibrium analysis of multiplex families of European descent from the British Diabetes Association (BDA) Warren Repository indicated that the association was stronger in -1123G > C compared to +1858C > T. In conclusion, the type 1 diabetes association with PTPN22 is confirmed, but it cannot be attributed solely to the +1858C > T variant. The promoter -1123G > C SNP is a more likely causative variant in PTPN22. PMID- 16470601 TI - Salmonella typhimurium specifically chemotax and proliferate in heterogeneous tumor tissue in vitro. AB - Multi-drug resistance greatly limits the efficacy of conventional blood-born chemotherapeutics, which have limited ability to penetrate tumor tissue and are ineffective at killing quiescent cells far from tumor vasculature. Nonpathogenic, motile bacteria can overcome both of theses limitations. We hypothesize that the accumulation of S. typhimurium in tumors is controlled by two mechanisms: (1) chemotaxis towards compounds produced by quiescent cancer cells and (2) preferential growth within tumor tissue. We tested this hypothesis by quantifying the relative contributions of these mechanisms using the tumor cylindroid model, which mimics the microenvironments of in vivo tumors. Time-lapse fluorescence microscopy was used to measure the accumulation of GFP-labeled S. typhimurium into cylindroids of different size. Cylindroids larger than 500 microm in diameter contain quiescent cells, whereas cylindroids smaller than 500 microm do not. Spatio-temporal profiles of bacterial concentration were fit to a mathematical model to calculate two parameters that describe bacterial interaction with tumors: intratumoral bacterial growth, M, and intratumoral bacterial chemoattraction, K. It was observed that S. typhimurium is attracted to cylindroids and accumulate at long time points in the central region of large cylindroids. Both intratumoral bacterial growth and chemotaxis were significantly greater in large cylindroids, suggesting that quiescent cells secrete bacterial chemoattractants and the presence of necrotic and quiescent cells enable S. typhimurium to replicate in tumor tissue. In this study, several mechanisms of S. typhimurium accumulation in solid tumors have been quantified, which we believe is an important step in the development of bacterial-based therapeutics to target tumor quiescence. PMID- 16470602 TI - Simultaneous control of turbidity and dilution rate through adjustment of medium composition in semi-continuous Chlamydomonas cultures. AB - For production of starch in algal cultures, a growth rate limited by a nutrient is an important factor. Under phototrophic conditions, turbidity must be also paid attention, as the shading effect may affect its productivity. Semi continuous cultivation methods, which enable control of turbidity and dilution rate (D) at the same time, have been developed for evaluation of those factors on starch production in Chlamydomonas sp. A specific feature of the methods is in a process of alternately feeding medium adjusted at two different nitrogen (N) concentrations. In the turbidostat-based method, a turbidostat culture was operated repeating three steps of determining D within a preset interval, alternating media by comparing the D with a preset value, and adjusting D in the next interval by feeding the selected medium. In the chemostat-based method, turbidity of a chemostat culture was controlled by repeating two steps of alternating media by comparing transmitted photon flux intensity (I) with a preset value and adjusting I by feeding the selected medium. D controlled by the turbidostat-based method reached quickly a preset value as low as 0.010/h, and then it was dispersed around but above the preset value. On the other hand, mean N concentrations of fed media formed a plateau. In the chemostat-based method, I was well controlled to a preset value while the mean N concentrations were a bit fluctuated. Starch concentration varied from 0.052 to 0.41 g/L with turbidity and D defined by these methods. PMID- 16470604 TI - Weekly cisplatin plus infusional high-dose 5-fluorouracil and leucovorin (P-HDFL) for metastatic urothelial carcinoma: an effective regimen with low toxicity. AB - BACKGROUND: Conventional systemic chemotherapy for metastatic urothelial carcinoma (UC) such as methotrexate, vinblastine, doxorubicin, and cisplatin (M VAC) or cisplatin, methotrexate, and vinblastine (CMV) is associated with significant dose-limiting toxicities and even treatment-related death. The authors developed a regimen that was designed to maintain efficacy, while reducing toxicities. METHODS: Between January 1998 and July 2003, 35 patients (median age, 71 yrs) with metastatic UC were treated with 4-week cycles of P-HDFL (cisplatin 35 mg/m(2), high-dose 5-fluorouracil [5-FU] 2,600 mg/m(2), and leucovorin 300 mg/m(2), on Days 1 and 8, all given by 24-hr infusion). On Day 15, only HDFL was given again. RESULTS: Among the 32 patients treated with > or = 2 cycles, 9 (28.1%) and 11 (34.4%) were complete and partial responders, respectively, with an overall response rate of 62.5% (95% confidence interval [CI], 45.9-79.2%). The median overall and progression-free survival was 12.3 months (95% CI, 8.2-16.4 mos) and 10.5 months (95% CI, 8.4-12.6 mos), respectively. Toxicity in a total of 121 courses (mean, 3.5 per patient) was modest, with WHO Grade 3 or 4 leukopenia and thrombocytopenia noted in only 1 and 0 patients, respectively. Grade 3 or 4 nausea, vomiting, mucositis, and diarrhea were noted in 3, 2, 0, and 2 patients, respectively. In general, patients tolerated the regimen very well. CONCLUSIONS: P-HDFL is a moderately active and considerably low-toxic regimen for metastatic UC. The excellent toxicity profile makes it a viable option for patients with poor general conditions. To reach any conclusion, randomized trials comparing P-HDFL with traditional cisplatin-based regimens are necessary. PMID- 16470603 TI - Calcium-responsive contractility during fertilization in sea urchin eggs. AB - Fertilization triggers a reorganization of oocyte cytoskeleton, and in sea urchins, there is a dramatic increase in cortical F-actin. However, the role that myosin II plays during fertilization remains largely unexplored. Myosin II is localized to the cortical cytoskeleton both before and after fertilization and to examine myosin II contractility in living cells, Lytechinus pictus eggs were observed by time-lapse microscopy. Upon sperm binding, a cell surface deflection traversed the egg that was followed by and dependent on the calcium wave. The calcium-dependence of surface contractility could be reproduced in unfertilized eggs, where mobilization of intracellular calcium in unfertilized eggs under compression resulted in a marked contractile response. Lastly, inhibition of myosin II delayed absorption of the fertilization cone, suggesting that myosin II not only responds to the same signals that activate eggs but also participates in the remodeling of the cortical actomyosin cytoskeleton during the first zygotic cell cycle. PMID- 16470605 TI - Poorly differentiated thyroid carcinomas defined on the basis of mitosis and necrosis: a clinicopathologic study of 58 patients. AB - BACKGROUND: Poorly differentiated thyroid carcinomas (PDTC) occupy an intermediate position at the prognostic level on the spectrum of thyroid carcinoma progression. However, their histologic definition is controversial. The objective of the current study was to assess the prognostic significance of PDTC defined on the basis of mitosis and necrosis and search for prognostic markers within this group of tumors that are predictive of overall survival (OS) and progression-free survival (PFS). METHODS: PDTC was defined as thyroid carcinoma with follicular cell differentiation at the histologic and/or immunohistochemical levels and displaying tumor necrosis and/or > or = 5 mitoses per 10 high-power fields (x400). Retrospective chart review and microscopic examination identified 58 patients with primary tumors meeting the above criteria and seen at the Memorial Sloan-Kettering Cancer Center between 1992 and 2004. These 58 patients were analyzed for various histologic, clinical, and imaging parameters. Each parameter was correlated with OS and PFS. RESULTS: Of the 58 patients studied, 22 (38%) patients died of disease with a 5-year OS rate of 60%. Forty-three of the 58 patients (74%) developed disease recurrence or disease progression, with a 5 year PFS rate of 25%. The median follow-up for the entire patient population was 42.6 months (range, 4-205 mos). A tumor size > 4 cm was found to be correlated with a decreased PFS time (P < 0.001). Those tumors with a capsule demonstrated a significantly improved OS compared with unencapsulated tumors (P = 0.001). The extent of capsular invasion was found to be a significant adverse factor for PFS (P = 0.05). The presence of extrathyroid extension into perithyroid soft tissue was found to be correlated with a decreased OS (P = 0.001) and PFS (P = 0.004). Of 27 patients with distant metastasis, 19 (70%) had concentrated radioactive iodine (RAI) at their metastatic sites. On multivariate analysis, extrathyroid extension and tumor size emerged as the only significant variables in predicting PFS (P = 0.04 and P = 0.01, respectively) whereas extrathyroid extension was found to be the sole independent prognostic factor for OS (P = 0.01). Growth pattern and cell type did not appear to influence outcome. CONCLUSIONS: PDTC defined on the basis of mitosis and necrosis constitutes a group of tumors that is more aggressive and homogeneous than PDTC defined by growth pattern. Within this group of patients, microstaging (tumor size, the extent of capsular invasion, and, especially, extrathyroid extension), and not growth pattern or cell type, is able to stratify patients into different prognostic categories. RAI uptake occurs in a significant number of patients with PDTC. PMID- 16470606 TI - Extended follow-up of a phase II trial in relapsed, refractory multiple myeloma:: final time-to-event results from the SUMMIT trial. AB - BACKGROUND: Bortezomib, a first-in-class proteasome inhibitor, has shown clinical activity in relapsed, refractory multiple myeloma in a pivotal Phase II trial, SUMMIT. METHODS: Patients received bortezomib 1.3 mg/m(2) on Days 1, 4, 8, and 11 followed by a 10-day rest period for up to 8 cycles. Dexamethasone 20 mg on the day of and the day after bortezomib was permitted for suboptimal response. Extended treatment beyond 8 cycles was offered to patients whose physicians felt they would benefit from additional therapy. Follow-up was conducted in all patients for a median of 23 months, an additional 13 months from the original report. RESULTS: Of 202 patients enrolled in SUMMIT, 193 were evaluable for response. Seven (4%) patients achieved a complete response, 12 (6%) achieved a nearly complete response, 34 (18%) achieved a partial response, and 14 (7%) had a minimal response while on bortezomib. The updated median duration of response to bortezomib alone was 12.7 months. The median overall time to progression for all SUMMIT patients was 7 months. For responding patients, the median time to progression was 13.9 months, whereas for those with progressive disease (PD) or who were not evaluable, the median time to progression was 1.3 months. The median overall survival (OS) for all SUMMIT patients was 17.0 months. Whereas the median OS for patients with PD or who were not evaluable was 8 months, the median OS for responding patients was not reached at 23 months of follow-up. CONCLUSIONS: These data demonstrate that treatment with bortezomib results in meaningful long-term benefit for patients with relapsed and refractory myeloma. PMID- 16470607 TI - Lamivudine prophylaxis reduces the incidence and severity of hepatitis in hepatitis B virus carriers who receive chemotherapy for lymphoma. AB - BACKGROUND: Hepatitis B virus (HBV) infection is a common disease in China. Severe hepatitis is a well recognized complication in HBV carriers with malignant disease who receive cytotoxic chemotherapy. The objective of the current study was to assess the value of antiviral lamivudine for reducing the incidence and severity of hepatitis in HBV carriers with lymphoma who receive chemotherapy. METHODS: Two groups were compared in this nonrandomized study. The prophylactic lamivudine group was comprised of 40 patients who received oral lamivudine at a dose of 100 mg daily before and until at least 8 weeks after they discontinued chemotherapy. The historic control group was comprised of 116 patients who received chemotherapy without lamivudine. The incidence and severity of hepatitis and other adverse clinical outcomes were compared between the two groups. Significant prognostic factors for the development of hepatitis were determined based on data derived from the control group. RESULTS: The two groups were comparable in most clinical baseline characteristics, including gender distribution, age, tumor types, primary treatment, hepatitis Be antigen status, and the use of anthracyclines or/and prednisone. In the prophylactic lamivudine group, there was significantly less incidence of hepatitis (17.5% vs. 51.7% in the control group; P = 0.000); less severe hepatitis (according to World Health Organization [WHO] criteria) (10% with Grade 1, 5% with Grade 2, and 2.5% with Grade 3 hepatitis vs. 3.4% with Grade 1, 12.1% with Grade 2, 12.9% with Grade 3, and 23.3% with Grade 4 hepatitis in the control group; P = 0.000); and less disruption of chemotherapy (10.0% vs. 37.1% in the control group; P = 0.001). The overall mortality as a result of hepatitis in the prophylactic lamivudine group was lower compared with that in the control group, but the difference was not statistically significant (0.0% vs. 5.2%; P = 0.163). In the control group, the factor associated with a greater risk of developing hepatitis was the use of prednisone. In the prophylactic lamivudine group, 1 of 40 patients (2.5%) developed hepatitis that was attributable to HBV reactivation. Further examination demonstrated that this single patient had a variation of HBV with YMDD mutations after the use of lamivudine for 9.2 months. CONCLUSIONS: The results of the current study confirmed previous reports that lamivudine prophylaxis significantly reduced the incidence and severity of hepatitis in HBV carriers who were receiving chemotherapy for lymphoma. The chemotherapy disruption rate as a result of severe hepatitis also was decreased significantly. PMID- 16470608 TI - Survival rates and patterns of care for patients diagnosed with supratentorial low-grade gliomas: data from the SEER program, 1973-2001. AB - BACKGROUND: Detailed population-based estimates of long-term survival as well as patterns of care for patients with low-grade gliomas examined by age at diagnosis, gender, and race have not been widely available. METHODS: Time to death was examined among 2009 individuals diagnosed with a supratentorial low grade glioma and reported to the Surveillance, Epidemiology, and End Results (SEER) program of the National Cancer Institute from 1973-2001 using Kaplan-Meier estimation. A Cox proportional hazards model was used to assess the effect of age at diagnosis, race, gender, histology, anatomic location within the brain, first course of treatment, and year of diagnosis upon this risk. RESULTS: The cumulative 5-, 10-, 15- and 20-year survival rates among all individuals initially diagnosed with a supratentorial low-grade glioma were 59.9% (95% confidence interval [95% CI], 57.6-62.2); 42.6% (95% CI, 39.9-45.2); 31.9% (95% CI, 29.0-34.8); and 26.0% (95% CI, 22.7-29.2), respectively. Improved survival was significantly associated with female gender (hazard ratio [HR], 0.84; 95% CI, 0.74-0.95), younger age, white race (HR, 0.70; 95% CI, 0.54-0.93), histology, and later year of diagnosis. Surgical treatment was associated with increased survival. The use of radiation therapy as a first course of treatment for these lesions has significantly decreased over time with the majority of patients receiving only surgery as a first course of treatment. CONCLUSIONS: Data for patients diagnosed with low-grade gliomas revealed increasing survival times over the past 25 years with a subset of patients surviving for decades. Differences in survival by race, gender, histology, and first course of treatment were appreciated. These data suggested that the clinical course of low-grade glioma for some patients may be more encouraging than previously perceived and that the identification of this group of patients may allow refinement of current treatment protocols. PMID- 16470609 TI - Interstitial 125I radiosurgery of supratentorial de novo WHO Grade 2 astrocytoma and oligoastrocytoma in adults: long-term results and prognostic factors. AB - BACKGROUND: Detailed long-term outcome data are not available for adult patients with World Health Organization (WHO) Grade 2 astrocytoma or oligoastrocytoma. METHODS: A previously published short-term data set of 239 adult patients with circumscribed de novo supratentorial astrocytoma (187 patients) and oligoastrocytoma (52 patients) treated with interstitial iodine-125 ((125)I) radiosurgery as primary treatment (1979-1992) was revisited. Survival, progression-free survival, functionally independent survival, postrecurrence survival, and time to malignant transformation were estimated with the Kaplan Meier method. Prognostic factors were obtained from the Cox multivariate proportional hazards model. RESULTS: Five-, 10-, and 15-year survival was 56%, 37%, and 26%, respectively (median follow-up, 10.3 yrs). Progression-free survival was 45%, 21%, and 14%, respectively. The corresponding malignant transformation rates were 33%, 54%, and 67%. No leveling off of the Kaplan-Meier curves could be observed for any of the chosen endpoints. Age > 50 years, a tumor volume > 20 mL, and/or a Karnofsky score < or = 80 were associated with decreased survival or progression-free survival. Age > 35 years and/or a tumor volume > 20 mL increased risk of malignant transformation. Prognostic factors determined subsets of patients with 10-year survival ranging from as low as 6% to as high as 55% and progression-free survival ranging 1-31%. CONCLUSIONS: Long-term tumor stabilization is rare. As outcome is mainly determined by treatment-independent factors, minimization of any treatment-related risk must be considered essential. PMID- 16470610 TI - Factors associated with surgical options for breast carcinoma. AB - BACKGROUND: Breast conservation surgery (BCS) and mastectomy have equivalent survival outcomes for women with breast carcinoma, but treatment decisions are affected by many factors. The current study evaluated the impact of patient and physician factors on surgical decision-making. METHODS: Statistical analyses were performed on a prospective multicenter study of patients with invasive breast carcinoma. Patient, physician, and geographic factors were considered. RESULTS: Of 4086 patients, BCS was performed in 2762 (67.6%) and mastectomy was performed in 1324 (32.4%). The median tumor size was 1.5 cm (range, < 0.1-9.0 cm) in patients undergoing BCS and 1.9 cm (range, 0.1-11.0 cm) in patients undergoing mastectomy (P < 0.00001). The median age of patients undergoing BCS was 59 years (range, 27-100 yrs), whereas patients who underwent mastectomy were older (median age of 63 yrs, range, 27-96 yrs [P < 0.00001]). Physicians in academic practices performed more lumpectomies than those who were not in an academic practice (70.9% vs. 65.7%; P = 0.001). More breast conservation procedures were performed by surgeons with a higher percentage of breast practice (P = 0.012). Geographic location was found to be significant, with the Northeast having the highest rate of breast conservation (70.8%) and the Southeast having the lowest (63.2%; P = 0.002). On multivariate analysis, patient age (odds ratio [OR]: 1.455; 95% confidence interval [95% CI], 1.247-1.699 [P < 0.001]), tumor size (P < 0.001), tumor palpability (OR: 0.613; 95% CI, 0.524-0.716 [P < 0.001]), histologic subtype (P = 0.018), tumor location in the breast (P < 0.001), physician academic affiliation (OR: 1.193; 95% CI: 1.021-1.393 [P = 0.026]), and geographic location (P = 0.045) were found to be significant. CONCLUSIONS: Treatment decisions were found to be related to patient clinicopathologic features, surgeon academic affiliation, and geographic location. Future studies will elucidate the communication and psychosocial factors that may influence patient decision making. PMID- 16470611 TI - Psychotherapy for patients with borderline personality disorder: focusing on the mechanisms of change. AB - A major development in the field of psychotherapy research is the growing recognition of the need for evidence on the mechanisms of change in psychotherapy. The empirical evidence that psychotherapy has a positive and significant effect must be amplified with data on the mechanisms of action in the various psychotherapies. This special issue is devoted to the articulation of putative mechanisms of change in the psychotherapy of patients with borderline personality disorder by leading researchers in this field. PMID- 16470612 TI - The mechanisms of change in the treatment of borderline personality disorder with transference focused psychotherapy. AB - We address how Transference Focused Psychotherapy (TFP) conceptualizes mechanisms in the cause and maintenance of borderline personality disorder (BPD) as well as change mechanisms both within the patient and in terms of specific therapists' interventions that engender patient change. Mechanisms of change at the level of the patient involve the integration of polarized representations of self and others; mechanisms of change at the level of the therapist's interventions include the structured treatment approach and the use of clarification, confrontation, and "transference" interpretations in the here and now of the therapeutic relationship. In addition, we briefly review evidence from our group regarding the following hypothesized mechanisms of change: contract setting, integration of representations, and changes in reflective functioning (RF) and affect regulation. PMID- 16470613 TI - Mouse Dach2 mutants do not exhibit gross defects in eye development or brain function. AB - Drosophila dachshund is a critical regulator of eye, brain, and limb formation. Vertebrate homologs, Dach1 and Dach2, are expressed in the developing retina, brain, and limbs, suggesting functional conservation of the dachshund/Dach gene family. Dach1 mutants die postnatally, but exhibit grossly normal development. Here we report the generation of Dach2 mutant mice. Although deletion of Dach2 exon 1 results in abrogation of RNA expression, Dach2 mutants are viable and fertile. Histochemical analysis reveals grossly normal Dach2 mutant eye development. In addition, a battery of neurological assays failed to yield significant differences in behavior between Dach2 mutants and controls. We discuss these findings in the light of published observations of DACH2 mutations in the human population. Finally, to test the functional conservation hypothesis, we generated Dach2; Dach1 double mutant mice. Dach double mutants die after birth, similar to Dach1 homozygotes. However, unlike Drosophila dachshund mutants that lack eyes and exhibit leg truncations, the eyes and limbs of Dach double mutants are present, suggesting differences between Dach and dachshund gene function during embryonic eye and limb formation. PMID- 16470614 TI - Visceral endoderm function is regulated by quaking and required for vascular development. AB - The quaking (qkI) gene produces three major alternatively spliced variants (qkI 5,-6,-7) that encode for proteins that share the RNA binding, KH domain. Previous studies utilizing the qk(k2) allele, which contains an N-ethyl-N-nitrosourea (ENU)-induced point mutation in the KH domain, demonstrate that this functional region of qkI is required for embryonic vascular development. In the current studies we demonstrate that qk(l-1)/qk(l-1) mutants, which lack the QKI-5 splice variant, also died at midgestation due to vascular remodeling defects. In addition, although all three QKI isoforms were expressed in the visceral endoderm of wildtype yolk sacs, qkI-6 and qkI-7 transcript and protein expression were suppressed in qk(k2)/qk(k2) and qk(l-1)/qk(l-1) mutant yolk sacs, suggesting that the KH-domain of QKI-5 was required for qkI-6 and qkI-7 expression. Further studies revealed that the cellular role of qkI is to regulate visceral endoderm function, including the local synthesis of retinoic acid (RA) and the subsequent control of endothelial cell proliferation, matrix production, and visceral endoderm survival. Although these defects were rescued by exogenous RA, visceral endoderm function or vascular remodeling were not restored. Thus, we conclude that qkI regulates visceral endoderm function, which is critical for vascular remodeling. PMID- 16470615 TI - The role of the forkhead transcription factor, Foxc1, in the development of the mouse lacrimal gland. AB - The lacrimal gland produces secretions that lubricate and protect the cornea of the eye. Foxc1 encodes a forkhead/winged helix transcription factor required for the development of many embryonic organs. Autosomal dominant mutations in human FOXC1 cause eye disorders such as Axenfeld-Rieger Syndrome and glaucoma iris hypoplasia, resulting from malformation of the anterior segment of the eye. We show here that lacrimal gland development is severely impaired in homozygous null Foxc1 mouse mutants, with reduced outgrowth and branching. Foxc1 is expressed in both the epithelium of the lacrimal gland and the surrounding mesenchyme. FGF10 stimulates the growth and branching morphogenesis in cultures of wild type and Foxc1 mutant gland epithelial buds. However, using micromass culture of lacrimal gland mesenchyme, we show that Bmp7 induces wild type mesenchyme cells to aggregate, but Foxc1 mutant cells do not respond. This study demonstrates that Foxc1 mediates the BMP signaling required for lacrimal gland development. PMID- 16470616 TI - Mouse embryonic stem cells are not susceptible to cytomegalovirus but acquire susceptibility during differentiation. AB - BACKGROUND: Cytomegalovirus (CMV) is the most significant infectious cause of congenital anomalies of the central nervous system caused by intrauterine infection in humans. The timing of infection and the susceptibility of cells in early gestational stages are not well understood. In this study we investigated the susceptibility of embryonic stem (ES) cells to CMV infection during differentiation. METHODS: ES cell lines were established from transgenic mice integrated with the murine CMV (MCMV) immediate-early (IE) promoter connected with a reporter lacZ gene. The susceptibility of the ES cells was analyzed in terms of viral gene expression and viral replication after induction of differentiation. RESULTS: ES cells were nonpermissive to MCMV infection in the undifferentiated state. Upon differentiation, permissive cells appeared approximately 2 weeks after the leukemia inhibitory factor was removed. Upon neural differentiation by retinoic acid (RA), glial cells showed specific susceptibility in terms of expression of the viral antigen. The MCMV IE promoter was not activated in ES cells from the transgenic mice. Activation of the IE promoter was detected approximately 2 weeks after induction of differentiation and observed predominantly in glial cells. Upon MCMV infection of the ES cells, viral infection was correlated with the activation of the IE promoter. CONCLUSIONS: ES cells are nonpermissive to MCMV infection and acquire permissiveness about 2 weeks after induction of differentiation, especially in glial cells. Acquisition of permissiveness in differentiated ES cells may be associated with activation of the IE promoter. PMID- 16470617 TI - Induction of parathyroid hormone-related peptide following peripheral nerve injury: role as a modulator of Schwann cell phenotype. AB - Parathyroid hormone-related peptide (PTHrP) is widely distributed in the rat nervous system, including the peripheral nervous system, where its function is unknown. PTHrP mRNA expression has recently been shown to be significantly elevated following axotomy of sympathetic ganglia, although the role of PTHrP was not investigated. The role of PTHrP in peripheral nerve injury was investigated in this study using the sciatic nerve injury model and dorsal root ganglion (DRG) explant model of nerve regeneration. We find that PTHrP is a constitutively secreted peptide of proliferating Schwann cells and that the PTHrP receptor (PTH1R) mRNA is expressed in isolated DRG and in sciatic nerve. Using the sciatic nerve injury model, we show that PTHrP is significantly upregulated in DRG and in sciatic nerve. In addition, in situ hybridization revealed significant localization of PTHrP mRNA to Schwann cells in the injured sciatic nerve. We also find that PTHrP causes a dramatic increase in the number of Schwann cells that align with and bundle regrowing axons in explants, characteristic of immature, dedifferentiated Schwann cells. In addition to stimulating migration of Schwann cells along the axonal membrane, PTHrP also stimulates migration on a type 1 collagen matrix. Furthermore, treatment of purified Schwann cell cultures with PTHrP results in the rapid phosphorylation of the cAMP response element protein, CREB. We propose that PTHrP acts by promoting the dedifferentiation of Schwann cells, a critical requirement for successful nerve regeneration and an effect consistent with known PTHrP functions in other cellular differentiation programs. PMID- 16470619 TI - Soluble and liquid-crystalline ovalenes. PMID- 16470622 TI - Hydrodynamic flow and electroosmotic flow in zirconia-packed capillaries. AB - Fused-silica capillaries were packed with Zirchrom-PBD stationary phase for application in CEC, nanoLC and pseudoelectrochromatography (PEC). Acido-basic properties of zirconia can be used to control the EOF even if the zirconia particles were coated by polybutadiene. As for native zirconia, the EOF is pH dependent and the pI is close to pH 5. The mixed-mode pressure-voltage technique induced a modulation of the mobile-phase velocity as well as an electrophoretic migration of the solutes in order to improve the resolution of the separation. A significant increase of the flow appeared when both hydrodynamic and EOFs were in the same direction. But an important reduction of the electroosmotic velocity was observed when the hydrodynamic flow and EOF were opposed in Zirchrom-PBD columns. This behaviour has been observed at high or low pH on several columns. Separations of neutral and charged compounds have been performed with these columns in PEC mode. PMID- 16470618 TI - Synthesis and characterization of metal-centered, six-membered, mixed-valent, heterometallic wheels of iron, manganese, and indium. AB - Heptanuclear metal-centered, six-membered, mixed-valent, heterometallic wheels 1 3 of iron, manganese, and indium were prepared in a one-pot reaction from N benzyldiethanolamine (H2L(1)), cesium carbonate, [PPh4]2[MnCl4], and FeCl3 or InCl3. All three complexes were characterized by the combination of elemental analysis, FAB mass spectroscopy, X-ray diffraction and cyclic voltammetry and in the case of 1 additionally by Mossbauer spectroscopy. In 1, four Mn(II) ions in the periphery are arranged in pairs alternating with one Fe(III) ion each, with an Fe(III) ion located in the center. In 2, three Mn(II) ions alternate with three In(III) ions, whereas in 3, four In(III) ions are arranged in pairs and alternate with one Mn(II) ion each. In 2 and 3 an Mn(II) ion is encapsulated in the center. PMID- 16470623 TI - Selection of two reliable parameters to evaluate the impact of the mobile-phase composition on capillary electrochromatography performance with monolithic and particle-packed capillary columns. AB - Different models have been described in the literature to evaluate the total porosity of CEC columns: gravimetric, flow as well as conductivity-based methods. In this study, these models have been compared for two kinds of CEC columns: two mixed-mode silica particle stationary phases and different monolithic columns (acrylate or polystyrene divinylbenzene-based). The total porosities measured from the conductivity-based methods were lower than the total column porosities obtained by gravimetric or flow methods for all the investigated columns while the wide distribution of observed values shows that conductivity-based methods discriminate columns more efficiently with very different properties. We propose a conductivity-based method taking into account the actual length proposed by Horvath, to evaluate what we call an "actual electrokinetic" porosity (AEP). This parameter, based on electrokinetic theory only, affords the most consistent evaluation of porosity under experimental CEC conditions for the packed- and acrylate-based monolithic columns. To illustrate the potential of AEP and actual EOF for the estimation of the performances of a CEC system (stationary and mobile phases) we studied the influence of the mobile-phase composition on these parameters for CEC separations with an ammonium embedded packed stationary phase. The AEP and the actual electroosmotic mobility should allow a better understanding of the perfusive EOF and stationary-phase wettability. For neutral compounds (substituted phenols), AEP evaluation allowed us to predict the mobile phase conditions able to enhance the efficiency while both AEP and actual EOF had to be considered in the case of peptide analysis. PMID- 16470624 TI - Rapid separation and determination of carbamate insecticides using isocratic elution pressurized capillary electrochromatography. AB - An isocratic elution pressurized CEC (pCEC) system was used to separate and determine ten carbamate insecticides. It was found that introduction of the electrical field, supplementary pressure, and SDS in the proposed method greatly improved the speed, column efficiency, selectivity, and repeatability for separation and determination of carbamates. On a capillary column of 75 microm ID packed with 3 microm octadecyl silica, baseline separation and detection of ten analytes was performed by using a mobile phase consisting of 30% v/v ACN and 70% v/v of 5 mmol/L ammonium acetate (pH 6.5) containing 1 mmol/L SDS and 0.01% triethylamine (TEA). Under the optimum conditions ten carbamate insecticides could be completely separated within 20 min. For the real vegetable samples, an SPE procedure for the cleanup of matrices was carried out prior to pCEC analysis. The detection limits of 0.05-1.6 mg/kg for ten carbamates and mean recoveries of 51.3-109.2% for eight kinds of vegetable samples at different concentrations of carbamates with RSD less than 11.4% were obtained, respectively. The proposed method has been proved to be effective in the rapid analysis of carbamate residues in vegetables. PMID- 16470625 TI - Micellar electrokinetic chromatography and capillary electrochromatography of nitroaromatic explosives in seawater. AB - The ability to separate nitroaromatic and nitramine explosives in seawater sample matrices is demonstrated using both MEKC and CEC. While several capillary-based separations exist for explosives, none address direct sampling from seawater, a sample matrix of particular interest in the detection of undersea mines. Direct comparisons are made between MEKC and CEC in terms of sensitivity and separation efficiency for the analysis of 14 explosives and explosive degradation products in seawater and diluted seawater. The use of high-salt stacking with MEKC results, on average, in a three-fold increase in the number of theoretical plates, and nearly double resolution for samples prepared in 25% seawater. By taking advantage of long injection times in conjunction with stacking, detection limits down to sub mg/L levels are attainable; however, resolution is sacrificed. CEC of explosive mixtures using sol-gels prepared from methyltrimethoxysilane does not perform as well as MEKC in terms of resolving power, but does permit extended injection times for concentrating analyte onto the head of the separation column with little or no subsequent loss in resolution. Electrokinetic injections of 8 min at high voltage allow for detection limits of explosives below 100 microg/L. PMID- 16470626 TI - Mixture design in the optimization of a microemulsion system for the electrokinetic chromatographic determination of ketorolac and its impurities: method development and validation. AB - Microemulsion EKC (MEEKC) was used for the determination of ketorolac and its three impurities. The microemulsion system was optimized, for the first time in the literature, using a multivariate strategy involving a mixture design. A 13 run experimental plan covering an experimental domain defined by the components aqueous phase (10 mM borate buffer pH 9.2), oil phase (n-heptane) and surfactant/cosurfactant (SDS/n-butanol) was carried out. Good results were obtained with all microemulsions tested considering as responses analysis time and resolution, and according to the desirability function the best microemulsion system was constituted by 90.0% 10 mM borate buffer, 2.0% n-heptane, 8.0% of SDS/n-butanol in 1:2 ratio. Finally, with the aim of reducing analysis time, a response surface study was carried out in the experimental domain defined by the process variables temperature and voltage and the best values were 17 degrees C and -17 kV, respectively. Applying the optimised conditions, a complete resolution among the analytes was obtained in about 3 min using the short-end injection method. The method was validated for both drug substances and drug product and was applied to the quality control of ketorolac in coated tablets. A comparison of MEEKC, MEKC and CEC for assaying ketorolac and its related substances has been made. PMID- 16470627 TI - Analysis of scleraxis and dermo-1 genes in a regenerating limb of Xenopus laevis. AB - Xenopus laevis larvae can regenerate an exact replica of the missing part of a limb after amputation at an early limb bud stage. However, this regenerative capacity gradually decreases during metamorphosis, and a froglet is only able to regenerate hypomorphic cartilage, resulting in a spike-like structure (spike). It has been reported that the spike has tissue deformities, e.g., a muscleless structure. However, our previous study demonstrated that the muscleless feature of the spike can be improved. The existence of other kinds of tissue, such as tendon, has not been clarified. In this study, we focused on the tendon and dermis, and we isolated the scleraxis and dermo-1 genes, which are known to be marker genes for the tendon and dermis, respectively. The expressions of these genes were investigated in both the developmental and regenerating processes of a Xenopus limb. Although muscle was needed to maintain scleraxis expression, scleraxis transcription was detectable in the muscleless spike. Additionally, although grafting of matured skin, including dermal tissue, inhibited limb regeneration, the expression of dermo-1, a dermal marker gene, was detected from the early stage of the froglet blastema. These results indicate that tendon precursor cells and dermal cells exist in the regenerating froglet blastema. Our results support the idea that spike formation in postmetamorphic Xenopus limbs is epimorphic regeneration. PMID- 16470628 TI - Expression of Xenopus laevis Lhx2 during eye development and evidence for divergent expression among vertebrates. AB - Members of the LIM homeodomain (LIM-HD) family of proteins are double zinc-finger containing transcription factors with important functions in pattern formation and cell lineage determination. The LIM-HD family member Lhx2 is required for normal eye, liver, and central nervous system formation. Lhx2(-/-) mice lack eyes, and experiments in Xenopus predict that Lhx2 forms a regulatory network with other eye field transcription factors to specify the eye field during eye formation. Here, we describe the structure and developmental expression pattern of the Xenopus laevis homologue, XLhx2. We show that XLhx2 shares significant amino acid sequence identity with other vertebrate Lhx2 proteins and Drosophila apterous (ap). The expression patterns of XLhx2 in the early neural plate and during eye development are consistent with a role in eye field specification and retinal differentiation. Despite highly similar expression patterns in the mouse and Xenopus central nervous system, divergent expression patterns were also observed. Phylogenetic analysis confirmed the identity of the isolated cDNA as a Xenopus ortholog of Lhx2. Therefore, in spite of structural similarities, the mouse and Xenopus Lhx2 expression patterns differ, suggesting potential functional differences in these species. PMID- 16470629 TI - Schema therapy for borderline personality disorder. AB - This article presents the Schema Therapy (Young, Klosko, & Weishaar, 2003) approach to the treatment of borderline personality disorder. Schema therapy draws on the cognitive-behavioral, attachment, psychodynamic, and emotion-focused traditions and conceptualizes patients who have borderline personality disorder as being under the sway of five modes or aspects of the self. The goal of the therapy is to reorganize this inner structure. To this end, there are four core mechanisms of change that are used in this therapy: (1) limited reparenting, (2) experiential imagery and dialogue work, (3) cognitive restructuring and education, and (4) behavioral pattern breaking. These interventions are used during the three phases of treatment: (1) bonding and emotional regulation, (2) schema mode change, and (3) development of autonomy. PMID- 16470630 TI - Proteomic analysis in human breast cancer: identification of a characteristic protein expression profile of malignant breast epithelium. AB - Gene expression analysis has become a promising tool in predicting the clinical course of malignant disease and the response to antineoplastic therapy. Surprisingly, only little is known about the protein expression pattern of human tumors. Recent advances in proteomic analysis allow proteins of interest to be identified by their expression and/or modification pattern in 2-DE rather than using the traditional approach of translating gene expression data. To identify a proteomic pattern that is characteristic for malignant breast epithelium, we performed differential 2-DE analysis in sets of microdissected malignant breast epithelia and corresponding adjacent normal breast epithelia from five patients with invasive breast carcinoma. Thirty-two protein spots were found to be selectively regulated in malignant epithelium, and were subjected to MALDI-TOF and/or immunoblotting for protein identification. Thirteen of the identified proteins had previously not been associated with breast cancer. The validity of these findings was confirmed by literature review and immunohistochemistry for identified proteins in an independent cohort of 50 breast cancer specimens. We here describe, for the first time, a proteomic analysis of matched normal and malignant epithelia from invasive breast carcinomas. This strategy leads to a better understanding of oncogenesis at an operational level and helps to characterize the malignant phenotype of individual tumors, and thereby to identify novel targets for antineoplastic therapy. PMID- 16470631 TI - Identification of novel phosphoproteins in signaling pathways triggered by latent membrane protein 1 using functional proteomics technology. AB - Previous studies have shown that the Epstein-Barr virus-encoded latent membrane protein1 (LMP1) could activate nuclear factor kappa B, activator protein-1, and Janus kinases/signal transducer and activation of transcription factors pathways. However, many signaling molecules and downstream target proteins triggered by LMP1 have not been identified. To determine the functional components in signaling pathways triggered by LMP1, we combined the novel strategy of phosphoprotein enrichment with proteomics technology to elucidate the signaling cascade activated by LMP1. We found that LMP1 could increase the quantity of total phosphoproteins by 18.03%, and 43 proteins showed significant changes in the degree of phosphorylation when LMP1 was expressed. Twenty-five signaling molecules or downstream targets of signaling pathways triggered by LMP1 were identified, several of which had previously been implicated in LMP1 signal pathways. The other proteins, including annexin A2, heat shock protein 27, stathmin, annexin I, basic transcription factor 3, and porin, were novel signaling molecules or targets with no previously known function in LMP1 signal transduction. The method used here has proven to be suitable for the identification of molecules involved in various signaling pathways. PMID- 16470632 TI - Capillary zone electrophoretic chiral discrimination using 6-O-(2-hydroxy-3 trimethylammoniopropyl)-beta-cyclodextrin as a chiral selector. AB - A charged highly water-soluble CD derivative, 6-O-(2-hydroxy-3 trimethylammoniopropyl)-beta-CD (herein noted as 6-HPTMA-beta-CD) was synthesized and successfully used as a chiral selector for enantiomeric separation of some acidic compounds by CZE in an uncoated capillary. Substitution with 2-hydroxy-3 trimethylammoniopropyl groups at the primary hydroxyl group of the CD was aimed at influencing the magnitude and selectivity of analyte-CD interactions. The behavior of 6-HPTMA-beta-CD was compared with that of the commercially available quaternary ammonium-beta-CD (QA-beta-CD) under the same separating conditions. The experiments were carried out using a BGE consisting of 50 mM phosphate in the pH range of 4-6 by adding a relatively low concentration of chiral selector (less than 10 mM). The effects of the concentration of CD and the pH of the electrolyte on the resolution of these compounds were studied. PMID- 16470633 TI - Chiral microemulsion electrokinetic chromatography with two chiral components: Improved separations via synergies between a chiral surfactant and a chiral cosurfactant. AB - In this study, the combination of two chiral components in a microemulsion formulation for the separation of enantiomers via microemulsion EKC (MEEKC) was successfully accomplished. Previous publications of chiral microemulsions have utilized only one chiral entity; the surfactant, cosurfactant, or oil was chiral. This is the first study, to date, of the effects of using two chiral species in a single pseudostationary phase (PSP). The chiral surfactant dodecoxycarbonylvaline (DDCV) was used in conjunction with the chiral cosurfactant S-2-hexanol. Ethyl acetate was incorporated as the oil core of the microemulsion and the buffer was 50 mM phosphate at a pH of 7. Additionally, a microemulsion prepared with racemic 2-hexanol was used for comparison to a previous DDCV microemulsion and as a baseline for the newly formulated dual chiral microemulsion. The efficiencies, resolutions, and enantioselectivities for the S-2-hexanol, racemic 2-hexanol, and original 1-butanol DDCV microemulsions are compared. The hexanol-based PSPs provide improved efficiencies and resolutions. To evaluate the combination of each DDCV enantiomer (R and S) with S-2-hexanol, changes in Gibb's free energy were calculated. A synergistic effect was found when two chiral components were combined to form a microemulsion. PMID- 16470634 TI - Enantiomeric screening of racemic citalopram and metabolites in human urine by entangled polymer solution capillary electrophoresis: an innovatory robustness/ruggedness study. AB - Several CE methods have been developed to achieve the chiral separation of citalopram (CIT) and its metabolites demethylcitalopram (DCIT), didemethylcitalopram (DDCIT), and citalopram N-oxide (CIT-NO). All of these compounds were present as racemic mixtures. The best method, which led to the first ever chiral screening of CIT, DCIT, DDCIT, and CIT-NO, involved the use of carboxymethyl-gamma-CD (CM-gamma-CD) and the entangled polymer hydroxypropylmethylcellulose (HPMC) as chiral and selectivity additives, respectively, in the buffer system. In an effort to improve the selectivity and sensitivity of the method, the chemical and instrumental parameters were optimized. The best conditions were short-end anodic hydrodynamic injection (6 s, 0.7 psi); as BGE pH 5, 20 mM phosphate buffer, 0.2% w/v CM-gamma-CD, 0.05% w/v HPMC; voltage of 28 kV with a ramp applied (0.4 s); cartridge temperature of 20 degrees C; detection at 205 nm. In addition, a simple and rapid achiral CE method for the determination of citalopram propionic acid (CIT-PA, the only anionic metabolite of CIT) is also reported for the first time. Prior to the electrophoretic procedure it was necessary to apply an extraction and preconcentration step to obtain analytes from the human urine samples. This was achieved using an optimized SPE process. Moreover, an innovatory experimental and statistical design approach, which involves the simultaneous evaluation of the global robustness and ruggedness effects, was applied. Both of the proposed methods proved to be very useful in the chiral pharmacokinetic screening of CIT and related metabolites in clinical human urine samples. PMID- 16470636 TI - Anti-obesity effects of green tea: from bedside to bench. AB - During the last decade, the traditional notion that green tea consumption benefits health has received significant scientific attention and, particularly, the areas of cardiovascular disease and cancer were subject to numerous studies. Due to the ever-growing obesity pandemic, the anti-obesity effects of green tea are being increasingly investigated in cell, animal, and human studies. Green tea, green tea catechins, and epigallocatechin gallate (EGCG) have been demonstrated in cell culture and animal models of obesity to reduce adipocyte differentiation and proliferation, lipogenesis, fat mass, body weight, fat absorption, plasma levels of triglycerides, free fatty acids, cholesterol, glucose, insulin and leptin, as well as to increase beta-oxidation and thermogenesis. Adipose tissue, liver, intestine, and skeletal muscle are target organs of green tea, mediating its anti-obesity effects. Studies conducted with human subjects report reduced body weight and body fat, as well as increased fat oxidation and thermogenesis and thereby confirm findings in cell culture systems and animal models of obesity. There is still a need for well-designed and controlled clinical studies to validate the existing and encouraging human studies. Since EGCG is regarded as the most active component of green tea, its specific effects on obesity should also be investigated in human trials. PMID- 16470637 TI - Green tea catechins as brain-permeable, natural iron chelators-antioxidants for the treatment of neurodegenerative disorders. AB - Neurodegeneration in Parkinson's, Alzheimer's, or other neurodegenerative diseases appears to be multifactorial, where a complex set of toxic reactions, including oxidative stress (OS), inflammation, reduced expression of trophic factors, and accumulation of protein aggregates, lead to the demise of neurons. One of the prominent pathological features is the abnormal accumulation of iron on top of the dying neurons and in the surrounding microglia. The capacity of free iron to enhance and promote the generation of toxic reactive oxygen radicals has been discussed numerous times. The observations that iron induces aggregation of inert alpha-synuclein and beta-amyloid peptides to toxic aggregates have reinforced the critical role of iron in OS-induced pathogenesis of neurodegeneration, supporting the notion that a combination of iron chelation and antioxidant therapy may be one significant approach for neuroprotection. Tea flavonoids (catechins) have been reported to possess divalent metal chelating, antioxidant, and anti-inflammatory activities, to penetrate the brain barrier and to protect neuronal death in a wide array of cellular and animal models of neurological diseases. This review aims to shed light on the multipharmacological neuroprotective activities of green tea catechins with special emphasis on their brain-permeable, nontoxic, transitional metal (iron and copper) chelatable/radical scavenger properties. PMID- 16470640 TI - Comparative proteome approach to characterize the high-pressure stress response of Lactobacillus sanfranciscensis DSM 20451(T). AB - High hydrostatic pressure (HHP) exerts diverse effects on microorganisms, leading to stress response and cell death. While inactivation of microorganisms by lethal HHP is well investigated in the context of food preservation and the hygienic safety of minimal food processes, sublethal HHP stress response and its effect on adaptation and cross-protection is less understood. In this study, the HHP stress response of Lactobacillus sanfranciscensis was characterized and compared with cold, heat, salt, acid and starvation stress at the proteome level by using 2-DE so as to provide insight into general versus specific stress responses. Sixteen proteins were found to be affected by HHP and were identified by using N-terminal amino acid sequencing and MS. Only one slightly increased protein was specific to the HHP response and showed homology to a clp protease. The other proteins were influenced by most of the investigated stresses in a similar way as HHP. The highest similarity in the HHP proteome was found to be with cold- and NaCl stressed cells, with 11 overlapping proteins. At the proteome level, L. sanfranciscensis appears to use overlapping subsets of stress-inducible proteins rather than stereotype responses. Our data suggest that a specific pressure response does not exist in this bacteria. PMID- 16470641 TI - Synthesis and biological evaluation of some hydroxypyrazole derivatives as anti inflammatory-antimicrobial agents. AB - Some hydroxypyrazole derivatives 2-7 were synthesized by cyclocondensation of the keto-ester 1 with hydrazines hydrate or substituted hydrazines followed by reduction and acylation with acetic anhydride or trifluoroacetic anhydride. The newly synthesized compounds were evaluated for their anti-inflammatory, antimicrobial activities. In addition, the ulcerogenic and acute toxicity profiles were determined. Compounds N-(4-(5-hydroxy-1-trifluoroacetyl-1H-pyrazol 3-yl)phenyl) trifluoroacetamide 4b, 3-(4-nitrophenyl)-1-(4-methoxyphenyl)-1H pyrazol-5-ol 5b, and N-(4-(5-hydroxy-1-methyl-1H-pyrazol-3 yl)phenyl)trifluoroacetamide 7b were proved to be the most active anti inflammatory, antimicrobial agents in the present study with a good safety margin and minimal or no ulcerogenic effect. PMID- 16470642 TI - Inhibitor and protein microarrays for activity-based recognition of lipolytic enzymes. AB - Protein and small-molecule microarrays are useful tools for high-throughput analysis of DNA-protein, protein-protein, and protein-small molecule interactions. Here we report on novel microarrays for activity screening of lipases and esterases based on phosphonic acid ester inhibitors. These compounds are activity recognition probes (ARPs) and bind to active serine hydrolases in a stoichiometric and irreversible manner. Protein microarrays were generated by spotting six different lipolytic enzymes onto hydrogel-coated glass slides. The activity of immobilized enzymes was determined after treatment with fluorescently labeled ARPs. Alternatively, biotinylated ARPs were bound to streptavidin slides in order to identify their affinity for enzymes in solution. Both systems, the protein- and ARP microarrays proved to be useful and versatile tools for the rapid identification and characterization of novel and known lipolytic enzymes. PMID- 16470643 TI - Conformational landscapes and free-jet rotational spectrum of indan-1-ol. PMID- 16470644 TI - On the protoporphyrin monolayers conformation. PMID- 16470646 TI - Tea and health. Interview with Yukihiko Hara by Fung-Lung Chung. PMID- 16470647 TI - Intracellular signaling network as a prime chemopreventive target of (-) epigallocatechin gallate. AB - Chemoprevention is an attempt to use either naturally occurring or synthetic substances or their mixtures to intervene in the progress of carcinogenesis. Recently, it has been shown that some edible phytochemicals alter gene expression, directly or indirectly, thereby regulating the carcinogenic processes. (-)-Epigallocatechin gallate (EGCG), a principal antioxidant derived from green tea, is one of the most extensively investigated chemopreventive phytochemicals. EGCG has been known to block each stage of carcinogenesis by modulating signal transduction pathways involved in cell proliferation, transformation, inflammation, apoptosis, metastasis and invasion. This review addresses the molecular target-based chemoprevention with EGCG by focusing on the common events mediated by transcription factors, such as NF-kappa B, activator protein-1 and nuclear factor erythroid 2 p45-related factor, and upstream kinases involved in the cellular signaling network. PMID- 16470648 TI - Tea, hormone-related cancers and endogenous hormone levels. AB - Tea is one of the most popular beverages consumed around the world, second only to water. There has been substantial interest in the potential role of tea in cancer prevention, particularly in respiratory and gastrointestinal tract cancers. Recent epidemiological data have linked tea intake to reduced risk of hormone-related cancers, including breast, ovarian and prostate cancers. Based on sparse data, there is suggestion that tea intake may influence circulating hormone levels, providing a plausible mechanism whereby tea intake may influence risk of hormone-related cancers. The major objectives of this paper are to review the epidemiological evidence on tea and risk of breast, ovarian, and prostate cancers as well as the human and non-human studies on tea and circulating hormone levels. We pay special attention to some of the limitations of the human studies and discuss future research needs. PMID- 16470649 TI - Synthesis and characterization of thiol containing furoxan derivatives as coligands for the preparation of potential bioreductive radiopharmaceuticals. AB - The synthesis and characterization of thiol-containing 1,2,5-oxadiazole N-oxide (TONO) derivatives and their use as monodentate coligands for the preparation of (99m)Tc complexes is presented. 3-Mercaptomethyl-4-phenyl-1,2,5-oxadiazol N(2) oxide and 3-(4-mercaptophenylmethylidenhydrazinocarbonyloxymethyl)-4-phenyl-1,2,5 oxadiazol N(2)-oxide were successfully synthesized and combined with the tridentate ligand N,N-bis(2-mercaptoethyl)-N',N'-diethylethylenediamine (BMEDA) to prepare "3+1 mixed ligand" technetium complexes. The( 99m)Tc complexes were obtained in high yield and radiochemical purity using low concentration of ligand and coligand. An alternative procedure using a xantate and a disulphide precursor of 3-mercaptomethyl-4-phenyl-1,2,5-oxadiazol N(2)-oxide yielded the same complex. Biological evaluation of the potentiality of the( 99m)Tc complexes as bioreductive radiopharmaceuticals was performed in normal CD1 mice and in mice bearing induced sarcoma. Tumour uptake was moderate but tumour/soft tissue ratio was favourable. Although these results are encouraging, further development is still necessary in order to achieve higher tumour uptake and lower gastrointestinal activity. PMID- 16470650 TI - Synthesis and potent antimicrobial activities of some novel retinoidal monocationic benzimidazoles. AB - Several 2-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthalen-2-yl)-1H benzimidazole-5-carboxamidine analogues were synthesized for their antibacterial and antifungal activities against S. aureus, Methicillin-resistant S. aureus (MRSA), C. albicans, and C. krusei. MIC values of the targeted compounds 43-58 are comparable to those of Fluconazole and Sultamicillin. The most potent compounds, 51 and 53, showed MIC values as 0.78 and 1.56 microg/mL against S. aureus and C. albicans, respectively. PMID- 16470651 TI - MapQuant: open-source software for large-scale protein quantification. AB - Whole-cell protein quantification using MS has proven to be a challenging task. Detection efficiency varies significantly from peptide to peptide, molecular identities are not evident a priori, and peptides are dispersed unevenly throughout the multidimensional data space. To overcome these challenges we developed an open-source software package, MapQuant, to quantify comprehensively organic species detected in large MS datasets. MapQuant treats an LC/MS experiment as an image and utilizes standard image processing techniques to perform noise filtering, watershed segmentation, peak finding, peak fitting, peak clustering, charge-state determination and carbon-content estimation. MapQuant reports abundance values that respond linearly with the amount of sample analyzed on both low- and high-resolution instruments (over a 1000-fold dynamic range). Background noise added to a sample, either as a medium-complexity peptide mixture or as a high-complexity trypsinized proteome, exerts negligible effects on the abundance values reported by MapQuant and with coefficients of variance comparable to other methods. Finally, MapQuant's ability to define accurate mass and retention time features of isotopic clusters on a high-resolution mass spectrometer can increase protein sequence coverage by assigning sequence identities to observed isotopic clusters without corresponding MS/MS data. PMID- 16470652 TI - Analysis of the interacting partners of the neuronal calcium-binding proteins L CaBP1, hippocalcin, NCS-1 and neurocalcin delta. AB - Intracellular Ca2+ signals are transduced by the binding of Ca2+ to sensor proteins, which subsequently modify the activity of their target proteins. Identification of these target proteins is, therefore, important for an understanding of cellular signalling processes. We have investigated the binding partners of four EF-hand Ca2+-binding proteins. Three proteins of the neuronal calcium sensor (NCS) family, hippocalcin, NCS-1 and neurocalcin delta were prepared as N-terminally tagged GST fusion proteins, and the less closely related protein L-CaBP1 was prepared in both N- and C-terminally tagged forms, the latter requiring generation of a new vector. Immobilised fusion proteins were used to purify binding partners from bovine brain cytosol and membrane extracts in the presence of 1 microM free Ca2+. Bound proteins were eluted with Ca2+-free and high-salt buffers and eluted proteins were identified by MALDI-MS and Western blotting. New protein targets detected included ARF1, Ca2+-dependent activator protein for secretion 1, cyclic nucleotide 3',5'-phosphodiesterase, the vacuolar ATPase, AP1 and AP2 complexes and the type I TGF-beta receptor. While certain of these interactions occurred with more than one of the Ca2+-binding proteins, others were found to be specific targets for particular Ca2+ sensors, and many of these did not overlap with known calmodulin-binding proteins. These findings provide new clues to the functional roles of the neuronal calcium sensor proteins. PMID- 16470653 TI - PRISM: a data management system for high-throughput proteomics. AB - Advanced proteomic research efforts involving areas such as systems biology or biomarker discovery are enabled by the use of high level informatics tools that allow the effective analysis of large quantities of differing types of data originating from various studies. Performing such analyses on a large scale is not feasible without a computational platform that performs data processing and management tasks. Such a platform must be able to provide high-throughput operation while having sufficient flexibility to accommodate evolving data analysis tools and methodologies. The Proteomics Research Information Storage and Management system (PRISM) provides a platform that serves the needs of the accurate mass and time tag approach developed at Pacific Northwest National Laboratory. PRISM incorporates a diverse set of analysis tools and allows a wide range of operations to be incorporated by using a state machine that is accessible to independent, distributed computational nodes. The system has scaled well as data volume has increased over several years, while allowing adaptability for incorporating new and improved data analysis tools for more effective proteomics research. PMID- 16470654 TI - Quantitation of protein phosphorylation in pregnant rat uteri using stable isotope dimethyl labeling coupled with IMAC. AB - Quantitative analysis of protein phosphorylation provides important insights into molecular signaling mechanisms and a better understanding of many cellular processes. In this study, we coupled stable isotope dimethyl labeling with immobilized metal affinity chromatography (IMAC) enrichment to quantify protein phosphorylation at MS-determined phosphorylation sites. The proposed method was first characterized using alpha- and beta-casein as two model phosphoproteins, and further applied to the analysis of pregnant rat uteri with and without treatment with 8-bromo-cGMP. Dimethyl labeling has several significant advantages: global, fast (within 5 min) and complete (near 100%). Our results indicate that the labeling has no adverse effect on the IMAC enrichment for tryptic peptides having single and multiple phosphorylation sites. Moreover, the enhanced a1 signal and the complete reaction by dimethyl labeling provide unequivocal identification of both the N-terminal amino acid and the number of the labeling site. Using these two criteria in data validation, which is particularly important for identifying phosphoproteins, we found that the confidence in interpreting dimethyl-labeled peptides had greatly increased. In the analysis of late gestation rat uteri, the abundance ratio between treated and un-treated phosphopeptide signals ranged from 0.51 to 1.69 with an average of around 1.01 +/- 0.25. The obtained ratio of the phosphorylation levels at Ser 15 of HSP27 was further confirmed by the consistent results obtained from Western blot analyses. Based on the analysis of the results, it is interesting to note that the activated cGMP dependent protein kinase G (PKG) seems to affect the phosphorylation of proteins associated with the inhibition of cell migration and proliferation, redistribution of actin-associated proteins, and the increase of protein synthesis in late-gestation uteri. These observations provide important evidence suggesting that activated PKG may play a critical role in the shift of pregnant uteri from proliferative to hypertrophic states. PMID- 16470655 TI - Comparative proteome analysis of Staphylococcus aureus biofilm and planktonic cells and correlation with transcriptome profiling. AB - Pathogenic staphylococci can form biofilms in which they show a higher resistance to antibiotics and the immune defense system than their planktonic counterparts, which suggests that the cells in a biofilm have an altered metabolic activity. Here, 2-D PAGE was used to identify secreted, cell wall-associated and cytoplasmic proteins expressed in Staphylococcus aureus after 8 and 48 h of growth. The proteins were separated at pH ranges of 4-7 or 6-11. The protein patterns revealed significant differences in 427 protein spots; from these, 258 non-redundant proteins were identified using ESI-MS/MS. Biofilm cells expressed higher levels of proteins associated with cell attachment and peptidoglycan synthesis, and in particular fibrinogen-binding proteins. Enzymes involved in pyruvate and formate metabolism were upregulated. Furthermore, biofilm cells expressed more staphylococcal accessory regulator A protein (SarA), which corroborates the positive effect of SarA on the expression of the intercellular adhesion operon ica and biofilm growth. In contrast, proteins, such as proteases and particularly immunodominant antigen A (IsaA) and staphylococcal secretory antigen (SsaA), were found in lower amounts. The RNA expression profiling largely supports the proteomic data. The results were mapped onto KEGG pathways. PMID- 16470656 TI - A proteomic analysis of 14-3-3 binding proteins from developing barley grains. AB - 14-3-3 proteins are important eukaryotic regulatory proteins. Barley (Hordeum vulgare L.) 14-3-3A was over-expressed, immobilised and used to affinity purify 14-3-3 binding proteins from developing barley grains. Binding was shown to be phosphorylation-dependent. These proteins were fractionated by PAGE and identified by MALDI-TOF MS. In total, 54 14-3-3 binding proteins were identified, 49 of these interactions are novel to plants. These proteins fell into a number of functional categories. The largest category was for carbohydrate metabolism, including plastidic enzymes for starch synthesis and modification. 14-3-3 was shown to be present in isolated plastids. Four of five enzymes involved in sucrose biosynthesis from triose phosphates were identified, suggesting co ordinated regulation of this pathway. Invertase and sucrose synthase, which break down sucrose to hexoses, were found. Sucrose synthase activity was shown to be inhibited by exogenous 14-3-3 in a dosage-dependent manner. The second-largest functional group was for proteins involved in stress and defence responses; for example, RGH2A, closely related to the MLA powdery mildew resistance protein, was found. This work illustrates the broad range of processes in which 14-3-3 may be involved, and augments previous data demonstrating key roles in carbohydrate metabolism and plant defence. PMID- 16470657 TI - Proteomic characterization of the effects of clofibrate on protein expression in rat liver. AB - Clofibrate is a peroxisome proliferator known to induce liver tumours in rats. A proteomics study was conducted to provide new insights into the molecular mechanisms of clofibrate-induced non-genotoxic hepatocarcinogenesis. Rats were treated with 250 mg/kg day clofibrate orally and sacrificed after 7 days. Proteins extracted from the liver were analysed by 2-DE using DIGE technology. The protein identification performed by MS showed that clofibrate induced up regulation of 77 proteins and down-regulation of 27 proteins. The highest expression ratios corresponded to proteins involved in a series of biochemical pathways such as lipid metabolism, fatty acid metabolism, amino acid metabolism, protein metabolism, citric acid cycle, xenobiotic detoxification and oxidative stress. Proteins implicated in cell proliferation and apoptosis, such as prohibitin, 10-formyl tetrahydrofolate dehydrogenase, senescence marker protein 30, pyridoxine 5'-phosphate oxidase and vimentin, were also identified as being regulated. These results provide leads for further investigations into the molecular mechanisms of liver tumours induced by clofibrate. In addition, MS results showed that a series of regulated proteins were detected as several spots corresponding to different pI and/or M(r). Differential effects on those variants could result from specific PTM and could be a specific molecular signature of the clofibrate-induced protein expression modulation in rat liver. PMID- 16470658 TI - Proteomic profiling of cell envelope-associated proteins from Staphylococcus aureus. AB - The emergence of highly virulent community acquired Staphylococcus aureus and continued progression of resistance to multiple antimicrobials, including methicillin and vancomycin, marks the reemergence of S. aureus as a serious health care threat. Investigation of proteins localized to the cell surface could help to elucidate mechanisms of virulence and antibiotic resistance in S. aureus. In this study, proteomic profiling methods were developed to solubilize, display, and evaluate abundance levels of proteins present in the supernatants of the lysostaphin-digested cell envelope from cultured vancomycin-intermediate S. aureus (VISA) cells. Combining approaches of 2-DE or chromatographic separation of proteins with MS analyses resulted in the identification of 144 proteins of particular interest. Of these proteins, 48 contained predicted cell wall localization or export signal motifs, including 14 with distinct covalent peptidoglycan-anchor sites, four of which are uncharacterized to date. One of the two most abundant cell envelope proteins, which showed remarkably high variations in MW and pI in the 2-DE gel display, was the S. aureus surface protein G. The display of numerous secreted proteins that are not covalently cell wall-anchored, suggests that, in the exponential growth phase, secreted proteins can be retained physiologically in the cell envelope and may interact with cell wall-anchored proteins and carbohydrate structures in a manner yet to be determined. The remaining 96 proteins, devoid of recognizable motifs, were repeatedly profiled in the VISA cell envelope fractions. We describe a novel semiquantitative method to determine abundance factors of such proteins in 2-DE gels of cell envelope fractions relative to whole cell lysates and discuss these data in the context of true cell envelope localization versus experimentally caused cell lysis. PMID- 16470659 TI - Surface-enhanced laser desorption/ionization time of flight mass spectrometry protein profiling identifies ubiquitin and ferritin light chain as prognostic biomarkers in node-negative breast cancer tumors. AB - Novel prognostic biomarkers are imperatively needed to help direct treatment decisions by typing subgroups of node-negative breast cancer patients. The current study has used a proteomic approach of SELDI-TOF-MS screening to identify differentially cytosolic expressed proteins with a prognostic impact in 30 node negative breast cancer patients with no relapse versus 30 patients with metastatic relapse. The data analysis took into account 73 peaks, among which 2 proved, by means of univariate Cox regression, to have a good cumulative prognostic-informative power. Repeated random sampling (n = 500) was performed to ensure the reliability of the peaks. Optimized thresholds were then computed to use both peaks as risk factors and, adding them to the St. Gallen ones, improve the prognostic classification of node-negative breast cancer patients. Identification of ubiquitin and ferritin light chain (FLC), corresponding to the two peaks of interest, was obtained using ProteinChip LDI-Qq-TOF-MS. Differential expression of the two proteins was further confirmed by Western blotting analyses and immunohistochemistry. SELDI-TOF-MS protein profiling clearly showed that a high level of cytosolic ubiquitin and/or a low level of FLC were associated with a good prognosis in breast cancer. PMID- 16470660 TI - Lipid particle composition of the yeast Yarrowia lipolytica depends on the carbon source. AB - Lipid particles (LP) of all types of cells are a depot of neutral lipids. The present investigation deals with the isolation of LP from the yeast Yarrowia lipolytica and the characterization of their lipid and protein composition. Properties of LP varied depending on the carbon source. LP from glucose-grown cells revealed a mean diameter of 650 nm with a hydrophobic core mainly formed of triacylglycerols (TAG) and a minor amount of steryl esters (SE). Oleic acid was the major fatty acid species esterified in LP. When cells were grown on oleic acid, LP size increased 3.8-fold, the particles exhibited a significantly lower ratio of TAG to SE, and the relative amount of oleic acid in LP lipids increased compared to cells grown on glucose. Analysis of LP proteins revealed an increasing number of polypeptides when cells were shifted from glucose- to oleic acid-containing medium. Twenty-one major LP proteins were identified under both growth conditions, and additional nine polypeptides were specific for growth on oleic acid. Identification of these proteins by MS and comparison of the deduced ORFs to those from Saccharomyces cerevisiae revealed that most proteins of Y. lipolytica LP are involved in lipid metabolism. LP proteins specific for growth on oleic acid are also enzymes involved in lipid metabolism, but some of them are also components of the intracellular traffic machinery. Thus, proteom analysis of LP proteins suggests involvement of this compartment in different cell biological processes. PMID- 16470663 TI - The identification and characterization of xenoantigenic nonhuman carbohydrate sequences in membrane proteins from porcine kidney. AB - The immunogenic nonhuman carbohydrate sequences in membrane proteins from porcine kidney were identified and characterized using MALDI-TOF MS and ESI-QTOF-MS. The MALDI profile, investigated by incubation with exoglycosidases, showed a series of about 40 carbohydrates that were identified as high mannose glycans (Man(3 9)GlcNAc2) and complex bi-, tri-, and tetra-antennary glycans with and without core fucose. The antennae of many of the complex glycans were terminated with alpha-galactose residues, with the numbers of these residues ranging from one up to the number of antennae. Negative ion ESI-MS/MS spectra confirmed the location of the alpha-galactose residues on the ends of the antennae. This total glycan profile of the membrane proteins from porcine kidney will thus provide important information for the study of molecular interactions between antigenic carbohydrates and proteins in xenotransplantation. PMID- 16470664 TI - Analysis of aromatic catabolic pathways in Pseudomonas putida KT 2440 using a combined proteomic approach: 2-DE/MS and cleavable isotope-coded affinity tag analysis. AB - Proteomic analysis of Pseudomonas putida KT2440 cultured in monocyclic aromatic compounds was performed using 2-DE/MS and cleavable isotope-coded affinity tag (ICAT) to determine whether proteins involved in aromatic compound degradation pathways were altered as predicted by genomic analysis (Jimenez et al., Environ Microbiol. 2002, 4, 824-841). Eighty unique proteins were identified by 2-DE/MS or MS/MS analysis from P. putida KT2440 cultured in the presence of six different organic compounds. Benzoate dioxygenase (BenA, BenD) and catechol 1,2-dioxygenase (CatA) were induced by benzoate. Protocatechuate 3,4-dixoygenase (PcaGH) was induced by p-hydroxybenzoate and vanilline. beta-Ketoadipyl CoA thiolase (PcaF) and 3-oxoadipate enol-lactone hydrolase (PcaD) were induced by benzoate, p hydroxybenzoate and vanilline, suggesting that benzoate, p-hydroxybenzoate and vanilline were degraded by different dioxygenases and then converged in the same beta-ketoadipate degradation pathway. An additional 110 proteins, including 19 proteins from 2-DE analysis, were identified by cleavable ICAT analysis for benzoate-induced proteomes, which complemented the 2-DE results. Phenylethylamine exposure induced beta-ketoacyl CoA thiolase (PhaD) and ring-opening enzyme (PhaL), both enzymes of the phenylacetate (pha) biodegradation pathway. Phenylalanine induced 4-hydroxyphenyl-pyruvate dioxygenase (Hpd) and homogentisate 1,2-dioxygenase (HmgA), key enzymes in the homogentisate degradation pathway. Alkyl hydroperoxide reductase (AphC) was induced under all aromatic compounds conditions. These results suggest that proteome analysis complements and supports predictive information obtained by genomic sequence analysis. PMID- 16470666 TI - A consensus procedure improving solvent accessibility prediction. AB - Prediction methods of structural features in 1D represent a useful tool for the understanding of folding, classification, and function of proteins, and, in particular, for 3D structure prediction. Among the structural aspects characterizing a protein, solvent accessibility has received great attention in recent years. The available methods proposed for predicting accessibility have never considered the combination of the results deriving from different methods to construct a consensus prediction able to provide more reliable results. A consensus approach that increases prediction accuracy using three high performance methods is described. The results of our method for three different protein data sets show that up to 3.0% improvement in prediction accuracy of solvent accessibility may be obtained by a consensus approach. The improvement also extends to the correlation coefficient. Application of our consensus approach to the accessibility prediction using only three prediction methods gives results better than single methods combined for consensus formation. Currently, the scarce availability of predictors with similar parameters defining solvent accessibility hinders the testing of other methods in our consensus procedure. PMID- 16470667 TI - An orbital localization criterion based on the theory of "fuzzy" atoms. AB - This work proposes a new procedure for localizing molecular and natural orbitals. The localization criterion presented here is based on the partitioning of the overlap matrix into atomic contributions within the theory of "fuzzy" atoms. Our approach has several advantages over other schemes: it is computationally inexpensive, preserves the sigma/pi-separability in planar systems and provides a straightforward interpretation of the resulting orbitals in terms of their localization indices and atomic occupancies. The corresponding algorithm has been implemented and its efficiency tested on selected molecular systems. PMID- 16470668 TI - Are calculated enthalpies of formation sometimes more reliable than experimental? A test on alkyl substituted benzoic acids. AB - Energies of 20 alkyl-substituted benzoic acids were calculated at the levels B3LYP/6-311+G(d,p)//B3LYP/6-311+G(d,p) and MP2/6-311+G(d,p)//MP2/6-311+G(d,p); the pertinent enthalpies at 298 K were calculated at the same levels. Comparison with experimental enthalpies of formation Delta(f)H degrees (g)(298) was carried out in terms of isodesmic reactions, that is, in the relative values. Of the four calculated quantities, the DFT enthalpies yielded best correlation with the standard deviation of 2.1 kJ mol(-1), near to the experimental uncertainty; the DFT energies are only slightly worse and the MP2 enthalpies or energies much worse. However, the DFT method overestimated systematically the substituent effects and had to be calibrated. Comparison with the experimental gas-phase acidities was less telling and the fit was worse because both methods overestimated the substituent effects. Extending the base in selected examples did not give better results. Although the systematic deviations are evidently due to the imperfections of the theoretical models, individual big deviations should be attributed to experimental errors or to the abnormal behavior of certain compounds at the experimental conditions. From this point of view, three examples of the so-called long-range effect claimed in the case of different benzoic acid derivatives, always for substituents in the meta position, must be refused as unproven because the experimental energies were not confirmed by calculations. PMID- 16470669 TI - Singular value decomposition of torsional angles of analogs of the dopamine reuptake inhibitor GBR 12909. AB - Analysis of large, flexible molecules, such as the dopamine reuptake inhibitor GBR 12909 (1), is complicated by the fact that they can take on a wide range of closely related conformations. The first step in the analysis is to classify the conformers into groups. Here, Singular Value Decomposition (SVD) was used to group conformations of GBR 12909 analogs by the similarity of their nonring torsional angles. The significance of the present work, the first application of SVD to the analysis of very flexible molecules, lies in the development of a novel scaling technique for circular data and in the grouping of molecular conformations using a technique that is independent of molecular alignment. Over 700 conformers each of a piperazine (2) and piperidine (3) analog of 1 were studied. Analysis of the score and loading plots showed that the conformers of 2 separate into three large groups due to torsional angles on the naphthalene side of the molecule, whereas those of 3 separate into nine groups due to torsional angles on the bisphenyl side of the molecule. These differences are due to nitrogen inversion at the unprotonated piperazinyl nitrogen of 2, which results in a different ensemble of conformers than those of 3, where no inversion is possible at the corresponding piperidinyl carbon. PMID- 16470670 TI - Monosilicon-substituted cyanoacetylene: a computational study. AB - A detailed theoretical investigation of the [H,Si,C(2),N] potential energy surfaces including 28 minimum isomers and 65 interconversion transition states is reported at the Gaussian-3//B3LYP/6-31G(d) level. Generally, the triplet species lie energetically higher than the singlet ones. The former three low-lying isomers are linear HCCNSi 1 (0.00 kcal/mol), branched SiC(H)CN 12 (7.09 kcal/mol), and bent HNCCSi 7 (14.22 kcal/mol), which are separated by rather high barriers from each other and are kinetically very stable with the least conversion barriers of 32.6-70.5 kcal/mol. Two energetically high-lying isomers HCNCSi 3 (42.99 kcal/mol) and SiC(H)NC 13 (36.05 kcal/mol) are also kinetically stable with a barrier of 49.19 and 21.42 kcal/mol, respectively. Additionally, five high-lying isomers, that is, three chainlike isomers, HCCSiN 2 (55.17), HCSiNC 6 (47.80), HSiNCC 11 (78.83), and one three-membered ring isomer HN-cSiCC 19 (51.21), and one four-membered ring isomer cSiCN(H)C 27 (50.6 kcal/mol), are predicted to each have lower conversion barriers of 12-18 kcal/mol and can be considered as meta-stable species. All of the predicted 10 isomers could exist as stable or meta-stable intermediates under suitable conditions. Finally, the structural and bonding analysis indicate that the [H,Si,C(2),N] molecule contains various properties that are of chemical interest (e.g., silylene, SiC triple bonding, and conjugate SiN triple bonding and CC triple bonding, charge-transfer specie, planar aromatic specie, cumulate double bonding). This is the first detailed theoretical study on the potential energy surfaces of the series of hydrogenated Si,C,C,N-containing molecules. The knowledge of the present monohydrogenated SiC(2)N isomerism could provide useful information for more highly hydrogenated or larger Si,C(2),N-containing species. PMID- 16470671 TI - Practical use of the International Olympic Committee Medical Commission Position Stand on the Female Athlete Triad: a case example. AB - The female athlete triad consists of the interrelated problems of disordered eating, amenorrhea, and osteoporosis, and it is believed to affect female athletes in all sports and at all levels of competition. OBJECTIVE: The current article highlights the Position Stand on the Female Athlete Triad of the International Olympic Committee's Medical Commission (IOCMC). METHOD: The literature related to disordered eating, energy availability, amenorrhea, and bone loss in athletes is briefly reviewed. A hypothetical case is presented to illustrate some of the common issues and problems encountered when working with athletes affected by the triad, such as the effect of weight on performance in "thin" sports, coach involvement, sport participation by symptomatic athletes, and treatment resistance/motivation. RESULTS: Strategies recommended by the position stand for managing those issues and problems are presented regarding the referral, evaluation, and treatment phases of the management process. CONCLUSION: Implications of the position stand are discussed in terms of the IOCMC's endorsement of the athlete's health being primary to her performance. PMID- 16470672 TI - Development of a liquid chromatography/tandem mass spectrometry method for the simultaneous determination of 16 mycotoxins on cellulose filters and in fungal cultures. AB - The development of a liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the simultaneous determination of 16 mycotoxins possibly related to the 'Sick Building Syndrome' on filters and in fungal cultures is described. Fungi-surface sampling as regards the 'Sick Building Syndrome' preferably happens by scraping off fungal material and vacuuming onto cellulose filters. Therefore, these two media were used as samples. They were spiked with nivalenol, deoxynivalenol, zearalenone, diacetoxyscirpenol, T-2 toxin, verrucarol, verrucarin A, neosolaniol, sterigmatocystin, roridin A, ochratoxin A, aflatoxin B1, aflatoxin B2, aflatoxin G1 and aflatoxin G2, which can be produced by isolates from fungi-damaged buildings. Deepoxy-deoxynivalenol was used as internal standard. Samples were extracted with organic solvents and the different mycotoxins were separated by high-performance liquid chromatography (HPLC) using a C18 reversed-phase SunFire analytical column and a mobile phase of variable mixtures of ammonium acetate (10 mM) and sodium acetate (20 microM) in water (solvent A) and in methanol (solvent B). The samples were run on-line with a Micromass Quattro Micro triple quadrupole mass spectrometer in positive electrospray ionisation mode using multiple reaction monitoring (MRM). The detection limits of the procedure varied from 50 to 0.009 pg/microL for filter samples and from 75 to 0.04 pg/microL for fungal culture samples. As the method includes few and non-labourious sample treatment steps, it should allow for a high throughput of samples. PMID- 16470673 TI - Stigmatic focusing reflectrons with spherical grids. PMID- 16470674 TI - Matrix-assisted laser desorption/ionization imaging mass spectrometry for direct measurement of clozapine in rat brain tissue. AB - Matrix-assisted laser desorption/ionization hyphenated with quadrupole time-of flight (QTOF) mass spectrometry (MS) has been used to directly determine the distribution of pharmaceuticals in rat brain tissue slices which might unravel their disposition for new drug development. Clozapine, an antipsychotic drug, and norclozapine were used as model compounds to investigate fundamental parameters such as matrix and solvent effects and irradiance dependence on MALDI intensity but also to address the issues with direct tissue imaging MS technique such as (1) uniform coating by the matrix, (2) linearity of MALDI signals, and (3) redistribution of surface analytes. The tissue sections were coated with various matrices on MALDI plates by airspray deposition prior to MS detection. MALDI signals of analytes were detected by monitoring the dissociation of the individual protonated molecules to their predominant MS/MS product ions. The matrices were chosen for tissue applications based on their ability to form a homogeneous coating of dense crystals and to yield greater sensitivity. Images revealing the spatial localization in tissue sections using MALDI-QTOF following a direct infusion of (3)H-clozapine into rat brain were found to be in good correlation with those using a radioautographic approach. The density of clozapine and its major metabolites from whole brain homogenates was further confirmed using fast high-performance liquid chromatography/tandem mass spectrometry (HPLC-MS/MS) procedures. PMID- 16470675 TI - Prenatal diagnosis of congenital epulis by three-dimensional ultrasound and magnetic resonance imaging. AB - Congenital epulis is a benign intraoral tumor that has rarely been diagnosed prenatally. We report a fetus with congenital epulis diagnosed by three dimensional (3-D) ultrasound and fetal magnetic resonance imaging (MRI) at 35 weeks' gestation. We show an image of congenital epulis using a new ultrasound technique, the 3-D multislice view, which shows images similar to CT or MRI. This is a short case report of congenital epulis diagnosed prenatally by 3-D ultrasound using multislice view and MRI. PMID- 16470676 TI - In vitro metabolism of a new cardioprotective agent, KR-32570, in human liver microsomes. AB - KR-32570 (5-(2-methoxy-5-chlorophenyl)furan-2-ylcarbonyl)guanidine) is a new reversible Na+/H+ exchanger inhibitor for preventing ischemia-reperfusion injury. This study was performed to identify the metabolic pathway of KR-32570 in human liver microsomes. Human liver microsomal incubation of KR-32570 in the presence of NADPH and UDPGA resulted in the formation of six metabolites, M1-M6. M1 was identified as O-desmethyl-KR-32570, on the basis of liquid chromatography/tandem mass spectrometric (LC/MS/MS) analysis with the synthesized authentic standard. M2 and M3 were suggested to be hydroxy-KR-32570 and hydroxy-O-desmethyl-KR-32570, respectively. M1, M2, and M3 were further metabolized to their glucuronide conjugates, M4, M5, and M6, respectively. In addition, the specific P450 isoforms responsible for KR-32570 oxidation to two major metabolites, O-desmethyl-KR-32570 and hydroxy-KR-32570, were identified using a combination of correlation analysis, chemical inhibition in human liver microsomes and metabolism by expressed recombinant P450 isoforms. The inhibitory potency of KR-32570 on clinically major P450s was investigated in human liver microsomes. The results show that CYP3A4 contributes to the oxidation of KR-32570 to hydroxy-KR-32570, and CYP1A2 play the predominant role in O-demethylation of KR-32570. KR-32570 was found to inhibit moderately the metabolism of CYP2C8 substrates. PMID- 16470677 TI - Fetal microchimerism is not involved in the pathogenesis of lichen sclerosus of the vulva. AB - OBJECTIVES: The aim of this study was to investigate a possible relationship between fetal cell microchimerism and lichen sclerosus of the vulva. We searched for the presence of male cells and DNA in vulval tissue samples. METHODS: Paraffin-embedded skin biopsy samples from 15 women affected with vulval lichen sclerosus who gave birth to at least one son were analyzed for the presence of microchimeric male cells using fluorescence in situ hybridization (FISH) and fluorescent PCR. We included three lichen sclerosus samples originating from women without male offspring, six vulval specimens without pathological finding originating from autopsies and seven male gingival specimens as controls. RESULTS: Nucleated cells containing Y-chromosome specific sequences were neither detected at any site of the lesions nor in normal vulval specimens by using FISH. These results were confirmed by the use of PCR amplification demonstrating only DNA sequences specific for the X chromosome. No female microchimerism was detected in the male gingival samples. CONCLUSION: Despite the limited number and size of the samples, we conclude that persistent male fetal cells are not involved in the pathogenesis of lichen sclerosus of the vulva, since we consistently could not detect Y-chromosome specific sequences by using two molecular techniques. PMID- 16470678 TI - Rapid characterization of edible oils by direct matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis using triacylglycerols. AB - Direct matrix-assisted laser desorption/ionization time-of-flight mass spectrometric (MALDI-TOFMS) analysis of solutions of edible fats/oils yielded spectra useful for their rapid differentiation and classification. Results also reflected the individual fatty acid components and their degree of unsaturation. After dissolution in hexane, MALDI-MS analysis revealed spectra showing characteristic triacylglycerols (TAGs), the main fat/oil components, as sodium adduct ions. The Euclidean distances calculated using the mass and intensity values for 20 TAGs were used to evaluate and compare spectra. With cluster analysis, animal fats grouped together differently than vegetable oils and the individual oils grouped together by type. The ion abundances for the individual TAGs and their presumed compositions were used to approximate the overall fatty acid composition of canola, soybean, corn, olive and peanut oil, as well as lard. Using this approach the calculated fatty acid compositions and degree of unsaturation generally fell within about 4% of literature values. When the degree of saturation was compared with values calculated from the package labeling the differences were about 7%. PMID- 16470679 TI - Investigating the in situ degradation of atrazine in groundwater. AB - This study focused on whether or not atrazine could be degraded by indigenous groundwater bacteria as part of an in situ remediation approach. Groundwater was taken from an unconfined middle upper chalk site where concentrations of atrazine and nitrate were typically in the ranges 0.02-0.2 microg litre-1 and 11.6-25.1 mg NO3-N litre-1 respectively. Sacrificial batch studies were performed using this groundwater spiked with atrazine at a concentration of 10 microg litre-1 in conjunction with a minimal mineral salts liquid (Glu-MMSL) medium which contained glucose as the sole carbon source. Treatments comprised either the Glu-MMSL groundwater cultured bacteria or Pseudomonas sp. strain ADP. Results from sacrificial batches indicated the occurrence of bacterial growth and denitrification as monitored by optical density (absorbance at 600 nm) and NO3-N content. Analysis of atrazine content by solid phase extraction coupled with high performance liquid chromatography showed no degradation of atrazine over a period of 103 days in either treatment. These results indicated that no acclimatised bacterial community featuring positive degraders to the herbicide atrazine had become established within this chalk aquifer in response to the trace levels encountered. PMID- 16470680 TI - The effect of insecticide application sequences on the control and insecticide resistance status of the peach-potato aphid, Myzus persicae (Hemiptera:Aphididae), on field crops of potato. AB - Experiments were done on commercial potato crops in the UK to investigate the effect of different insecticide sequences on the control and insecticide resistance status of Myzus persicae (Sulzer). The work was done to provide field validation of similar laboratory studies done in 'field simulators'. To ensure adequate aphid populations and to influence the initial resistance status of the aphid population, cultured M. persicae from a clone of known resistance status (esterase R1, kdr heterozygote, non-MACE (modified acetylcholinesterase)) were inoculated into both experiments. Two-spray programmes starting with lambda cyhalothrin (a pyrethroid insecticide) gave poor control in comparison with programmes starting with pirimicarb (a carbamate insecticide) or pirimicarb containing mixtures. This concurred closely with the results obtained from single applications in field simulator studies. Treatment sequences containing pymetrozine (a pyridine azomethine insecticide) were also effective, though slower-acting. This again concurs with field simulator studies. The proportions of aphids carrying different resistance mechanisms were largely unaffected by treatment in these experiments. The implications of these results for field control strategies are discussed. PMID- 16470681 TI - Improved efficacy of ethyl formate against stored grain insects by combination with carbon dioxide in a 'dynamic' application. AB - Ethyl formate is being evaluated as a fumigant for stored grain as it is a potential alternative to the ozone-depleting fumigant methyl bromide and to phosphine, which is under pressure owing to the development of strong resistance in stored grain insects. However, use of ethyl formate faces significant challenges, such as poor penetration through grain, significant losses to grain sorption, high concentrations of fumigant required to control insects, and flammability risks, which have limited its further development. In this study it was found that the combination of carbon dioxide (5-20%) with ethyl formate significantly enhanced efficacy of the fumigant against external living stages of the rice weevil, Sitophilus oryzae (L.), the lesser grain borer, Rhyzopertha dominica F., and the flour beetle, Tribolium castaneum (Herbst). Dynamic application of ethyl formate and carbon dioxide mixture (100 mg litre-1 ethyl formate, 20% CO2) pumped through a model silo containing wheat (50 kg) for one gas exchange was also investigated. A flow rate of 6 litres min-1 gave a relatively even distribution of fumigant throughout the grain column and similar mortality levels among cultures of S. oryzae and T. castaneum placed at three positions, the top, middle and bottom of the column. Mortality of 99.8% of mixed stage cultures of T. castaneum and 95.1% of S. oryzae was achieved in 3 h exposures to 111 and 185 mg ethyl formate h litre-1 respectively applied by the dynamic method. It is concluded that the combination of carbon dioxide with ethyl formate and dynamic application enhances distribution and efficacy of the fumigant against stored grain insects. PMID- 16470682 TI - Matrix-assisted laser desorption/ionization in-source decay combined with tandem time-of-flight mass spectrometry of permethylated oligosaccharides: targeted characterization of specific parts of the glycan structure. AB - Matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF-MS) has been introduced in recent years as a valuable tool for the structural characterization of permethylated oligosaccharides. In this report, we describe the combination of MALDI in-source decay (ISD) with the subsequent TOF/TOF-MS analyses of specific fragments, allowing the detailed characterization of the selected part of the oligosaccharide molecule. Part of the second-generation fragment ions were different from those observed in conventional MALDI-TOF/TOF-MS experiments. Other fragments, which had already been observed in conventional MALDI-TOF/TOF-MS and again showed up in second generation fragment analysis, could be assigned to specific parts of the molecule. Our approach disclosed different structural features of the oligosaccharides: due to permethylation, the glycosidic linkage fragments allowed the distinction between terminal, monosubstituted and disubstituted monosaccharides and indicated the oligosaccharide sequence. Moreover, substitution positions were deduced based on characteristic cross-ring fragmentation by high-energy collision-induced fragmentation. In conclusion, combination of MALDI-ISD with TOF/TOF-MS allows the detailed characterization of specific moieties of permethylated oligosaccharides and is, therefore, a powerful technique for structural glycomics. PMID- 16470683 TI - Use of oils combined with low doses of insecticide for the control of Myzus persicae and PVY epidemics. AB - Experiments were carried out in the laboratory to assess the insecticidal effect on Myzus persicae Sulzer of different oils applied alone or combined with imidacloprid or pirimicarb. The oils tested were a horticultural mineral oil, a refined rapeseed oil, a refined soya oil and a raw fish oil. When the oils were sprayed alone on pepper plants infested with M. persicae, mineral oil caused the highest mortality of aphids (over 80%). Applied before aphid infestation of pepper leaves and in mixture with low doses of imidacloprid (at one-fifth of the dose recommended by the manufacturer) and pirimicarb (at one-tenth of the dose recommended by the manufacturer), the oils did not significantly increase the toxicity of the insecticides alone. However, sprayed on aphid-infested pepper plants, the mortality rates achieved by imidacloprid/mineral oil and imidacloprid/rapeseed oil mixtures were significantly higher than those achieved by imidacloprid alone at 16 and 24 h. In a field experiment the effect on the incidence of the potato virus (PVY) of the oils in combination with imidacloprid was determined. Mineral oil, rapeseed oil and soya oil were sprayed eight times onto seed potato plants treated with imidacloprid before sowing. Mineral oil reduced PVY-infected plants by 60% and rapeseed oil by 40% compared with plots treated with imidacloprid. The oils applied as 10 ml litre-1 emulsions in water did not cause symptoms of phytotoxicity on the potato plants, and yield was not reduced. PMID- 16470684 TI - Supraspinal input is dispensable to generate glycine-mediated locomotive behaviors in the zebrafish embryo. AB - The anatomy of the developing zebrafish spinal cord is relatively simple but, despite this simplicity, it generates a sequence of three patterns of locomotive behaviors. The first behavior exhibited is spontaneous movement, then touch evoked coiling, and finally swimming. Previous studies in zebrafish have suggested that spontaneous movements occur independent of supraspinal input and do not require chemical neurotransmission, while touch-evoked coiling and swimming depend on glycinergic neurotransmission as well as supraspinal input. In contrast, studies in other vertebrate preparations have shown that spontaneous movement requires glycine and other neurotransmitters and that later behaviors do not require supraspinal input. Here, we use lesion analysis combined with high speed kinematic analysis to re-examine the role of glycine and supraspinal input in each of the three behaviors. We find that, similar to other vertebrate preparations, supraspinal input is not essential for spontaneous movement, touch evoked coiling, or swimming behavior. Moreover, we find that blockade of glycinergic neurotransmission decreases the rate of spontaneous movement and impairs touch-evoked coiling and swimming, suggesting that glycinergic neurotransmission plays critical yet distinct roles for individual patterns of locomotive behaviors. PMID- 16470685 TI - Effects of amyloid peptides on A-type K+ currents of Drosophila larval cholinergic neurons. AB - Accumulation of amyloid (Abeta) peptides has been suggested to be the primary event in Alzheimer's disease. In neurons, K+ channels regulate a number of processes, including setting the resting potential, keeping action potentials short, timing interspike intervals, synaptic plasticity, and cell death. In particular, A-type K+ channels have been implicated in the onset of LTP in mammalian neurons, which is thought to underlie learning and memory. A number of studies have shown that Abeta peptides alter the properties of K+ currents in mammalian neurons. We set out to determine the effects of Abeta peptides on the neuronal A-type K+ channels of Drosophila. Treatment of cells for 18 h with 1 microM Abeta1-42 altered the kinetics of the A-type K+ current, shifting steady state inactivation to more depolarized potentials and increasing the rate of recovery from inactivation. It also caused a decrease in neuronal viability. Thus it seems that alteration in the properties of the A-type K+ current is a prelude to the amyloid-induced death of neurons. This alteration in the properties of the A-type K+ current may provide a basis for the early memory impairment that was observed prior to neurodegeneration in a recent study of a transgenic Drosophila melanogaster line over-expressing the human Abeta1-42 peptide. PMID- 16470686 TI - Healthy 12-year-old boy with mosaic inv dup(15)(q13). PMID- 16470687 TI - Spondylo-ocular syndrome: a new entity involving the eye and spine. PMID- 16470688 TI - Variable phenotype and associations in chromosome 22q11.2 microdeletion. PMID- 16470689 TI - Interstitial 2.2 Mb deletion at 9q34 in a patient with mental retardation but without classical features of the 9q subtelomeric deletion syndrome. AB - In a female patient with mild mental retardation an interstitial subtelomeric 9q34.3 deletion was identified by a multiplex ligation-dependent probe amplification (MLPA) based screen for subtelomeric abnormalities. Further characterization of the deletion by high-resolution tiling path array-based comparative genomic hybridization (array CGH) revealed a size of 2.2 Mb. The woman lacked the typical 9qter deletion phenotype characteristics, which is inline with the finding that both Eu-HMTase1 (EHMT) genes were present. However, she presented with mild mental retardation, some mild facial dysmorphisms and aplasia cutis. This is another example of an interstitial subtelomeric deletion, which underscores that further characterizing the precise nature of the deletion is of clinical importance. Moreover, it confirms the importance of the Eu-HMTase1 gene as the major causative factor of the classical 9qter syndrome phenotype. PMID- 16470690 TI - A novel mutation in the DNA-binding domain of MAF at 16q23.1 associated with autosomal dominant "cerulean cataract" in an Indian family. AB - Congenital cataract, a clinically and genetically highly heterogeneous eye disorder, is one of the significant causes of visual impairment or blindness in children. It is frequently inherited as an autosomal dominant trait. We investigated a three-generation family of Indian origin with 12 members affected with cerulean cataract. Linkage analysis was carried out in this family using more than 100 microsatellite markers for the known cataract candidate gene loci. A positive two-point lod score of 3.9 at theta = 0.000, indicative of linkage, was obtained with three microsatellite markers for chromosome 16. Multipoint and haplotype analysis narrowed the cataract locus to a 15.3 cM region between markers D16S518 and D16S511 that corresponds to the region 16q23.1. Direct sequencing of the candidate gene MAF, which lies in the critical linked region, revealed a novel heterozygous missense mutation in the basic region (BR) of the DNA-binding domain. This sequence change was considered pathogenic as it segregated in all affected family members, neither seen in unaffected family members nor in 106 unrelated controls. The mutation also results in substitution of highly conserved lysine 297 by arginine (K297R) that affects a residue that forms a part of a predicted DNA-interaction region of the protein. The association of microcornea with congenital cataract in some affected individuals further underlines the role of the MAF transcription factor in lens and anterior ocular development. Our findings expand the mutation spectrum of MAF in association with congenital cataract and highlight the genetic and phenotypic heterogeneity of congenital cataract. PMID- 16470692 TI - Proton MRS of early post-natal mouse brain modifications in vivo. AB - NMR provides a non-invasive tool for the phenotypic characterisation of mouse models. The aim of the present study was to apply reliable in vivo MRS techniques for non-invasive investigations of brain development in normal and transgenic mice, by monitoring metabolite concentrations in different brain regions. The conditions of anaesthesia, immobilisation and respiratory monitoring were optimized to carry out in vivo MRS studies in young mice. All the experiments were performed in normal mice, at 9.4 T, applying a point-resolved spectroscopy (PRESS) sequence (TR = 2,000 ms; TE = 130 ms). We obtained reproducible in vivo (1)H NMR spectra of wild-type mouse brains as early as post-natal day 5, which allowed us to follow brain maturation variations from post-natal days 5 to 21. The survival rate of animals was between 66 and 90% at post-natal days 5 and 21, respectively. Developmental changes of metabolite concentrations were measured in three brain regions: the thalamus, a region rich in cell bodies, the olfactory bulb, rich in fibre tracts actively myelinated during brain maturation, and the cerebellum. The voxel size varied from 2 to 8 microL according to the size of the brain structure analysed. The absolute concentrations of the total creatine, taurine, total choline, N-acetylaspartate and of the glutamate/glutamine pool were determined from (1)H NMR spectra obtained in the different brain regions at post-natal day 5, 10, 15 and 21. Variations observed during brain development were in accordance with those previously reported in mice using ex vivo MRS studies, and also in rats and humans in vivo. Possibilities of longitudinal MRS analysis in maturing mice brains provide new perspectives to characterise better the tremendous number of transgenic mutant mice generated with the aim of decrypting the complexity of brain development and neurodegenerative diseases but also to follow the impact of environmental and therapeutic factors. PMID- 16470693 TI - Laboratory monitoring of potassium and creatinine in ambulatory patients receiving angiotensin converting enzyme inhibitors and angiotensin receptor blockers. AB - PURPOSE: Serum potassium and creatinine monitoring is recommended for patients prescribed angiotensin converting enzyme inhibitors (ACEi) and angiotensin receptor blockers (ARB). Much has been written about hyperkalemia associated with these drugs; little is known about laboratory monitoring patterns. The purpose of this retrospective cohort study was to assess creatinine and potassium monitoring and characteristics associated with monitoring among patients dispensed ACEi or ARB. METHODS: This study was conducted in 10 United States health maintenance organizations. Study patients (n = 52 906) were aged 18 or older with dispensings of ACEi or ARB for at least 1 year. Serum potassium and creatinine monitoring were assessed from administrative data and medical records. RESULTS: More than two-thirds (68.4%) of patients received laboratory monitoring. Likelihood of monitoring increased with age (adjusted odds ratio [OR] 2.10; 95% confidence interval [95%CI] 1.93, 2.28 [individuals >or= 80 compared to <50 years]), >9 outpatient visits (OR 1.46; 95%CI 1.39, 1.54), hospitalization (OR 1.15; 95%CI 1.06, 1.25), concomitant medications (potassium [OR 2.01; 95%CI 1.84, 2.20], diuretics [OR 1.54; 95%CI 1.47, 1.61], digoxin [OR 1.15; 95%CI 1.01, 1.30]), and comorbidities (diabetes [OR 1.68; 95%CI 1.61, 1.75], heart failure [OR 1.73; 95%CI 1.57, 1.90], chronic kidney disease [OR 2.95; 95%CI 2.48. 3.51]). CONCLUSIONS: Nearly one-third of patients dispensed ACEi or ARB did not undergo laboratory monitoring at least yearly. Though patients at increased risk of hyperkalemia were more likely to be monitored, many remained unmonitored. PMID- 16470694 TI - Primary palmar hyperhidrosis locus maps to 14q11.2-q13. AB - Primary palmar hyperhidrosis (PPH) is a unique disorder of unknown cause. It is characterized by excessive perspiration of the eccrine sweat gland in the palm, sole, and the axilla. It is presumed that PPH results from overactivation of the cholinergic sympathetic nerve or dysfunction of the autonomic nervous system. There have been no genetic studies on the disease. We performed a linkage analysis of 11 families including 42 affected and 40 unaffected members using genome-wide DNA polymorphic markers to identify the disease locus. Diagnosis of their PPH was made by direct inspection, interviewing and measurement of the sweating rate with perspirometer. Consequently, from data of three of the 11 families examined, the combined maximum two-point LOD scores of 3.08 and 3.16 (recombination fraction = 0) were obtained at the D14S283 and D14S264 loci, respectively, on chromosome 14q11.2-q13, under an assumption that two liability conditions depend on age. These regions were ruled out in eight other families. Haplotype analysis of the three families supported that one of the PPH locus is assigned at minimum to about a 6-cM interval between D14S1070 and D14S990 and at maximum to about a 30-cM interval between D14S1070 and D14S70. This is the first report of systemic mapping of the PPH locus. PMID- 16470696 TI - Mosaic trisomy 6 and maternal uniparental disomy 6 in a 23-week gestation fetus with atrioventricular septal defect. AB - Trisomy 6 is seen in early miscarriages in association with an intact, empty amniotic sac or as a pseudomosaic in amniotic fluid cultures. We report the finding of mosaic trisomy 6 in a 23-week-gestation pregnancy terminated because of intrauterine death. The post-mortem showed a well formed macerated male fetus with an atrioventricular septal defect and an exomphalos. By conventional cytogenetics, trisomy 6 was found in 12 out of 25 (48%) fibroblast colonies from fetal skin and 21 out of 32 (66%) colonies derived from amnion, while the remaining metaphases showed an apparently normal male karyotype. Molecular genetic studies on DNA from uncultured fetal skin and cord samples using polymorphic microsatellite repeat sequences showed no evidence of trisomy 6, but demonstrated that both chromosome 6 homologs were of maternal origin consistent with maternal uniparental disomy (UPD). PMID- 16470697 TI - Targeted comparative proteomics by liquid chromatography/matrix-assisted laser desorption/ionization triple-quadrupole mass spectrometry. AB - Here we report the first application of a matrix-assisted laser desorption/ionization (MALDI) triple-quadrupole mass spectrometer for targeted proteomics. Employing an amine-specific isotopic labelling approach, the technique was validated using five randomly selected bovine serum albumin peptides differentially labelled at known ratios. An indirect benefit of the isotopic labelling technique is a significant enhancement of the a1 ion in tandem mass (MS/MS) spectra of all peptides studied. Therefore, the a1 ion was selected as the fragment ion for multiple reaction monitoring (MRM) in all cases, eliminating tedious method development and optimization. Accurate quantification was achieved with an average relative standard deviation (RSD) of 5% (n = 5) and a detection limit of 14 amol. The technique was then applied to validate an important virulence biomarker of the fungal pathogen Candida albicans, which was not accurately quantified using global proteomics experiment employing two dimensional liquid chromatography/electrospray ionization tandem mass spectrometry (2D-LC/ESI)-MS/MS. Using LC/MALDI-MRM analysis of five tryptic peptides, the protein PHR1 was found to be upregulated in the hyphal (pathogenic) form of C. albicans by a factor of 7.7 +/- 0.8. PMID- 16470698 TI - Ring chromosome 4 and Wolf-Hirschhorn syndrome (WHS) in a child with multiple anomalies. AB - We report on a 16-month-old male patient with ring chromosome 4 and deletion of Wolf-Hirschhorn syndrome (WHS) region with multiple congenital anomalies including unilateral cleft lip and palate, iris coloboma, microcephaly, midgut malrotation, hypospadias, and double urethral orifices. Peripheral chromosome analysis of the patient showed 46,XY,r(4)(p16.3q35) de novo. Multicolor fluorescence in situ hybridization (FISH) study was also performed and according to multicolor banding (MCB) a r(4)(::p16.3 --> q34.3 approximately 35.1::) was found in all metaphases. Subtelomeric 4p region, subtelomeric 4q region, as well as, Wolf-Hirschhorn critical region were deleted in ring chromosome 4. Genomic microarray analysis was also performed to delineate the size of deletion. Cranial magnetic resonance imaging (MRI) showed hypoplastic corpus callosum, delayed myelinization, and frontal and occipital lobe atrophies. Both maternal and paternal chromosomal analyses were normal. We compare the phenotypic appearance of our patient with the previously reported 16 cases of ring chromosome 4 in the medical literature. PMID- 16470701 TI - A nonsense mutation of PEPD in four Amish children with prolidase deficiency. AB - Encoded by the peptidase D (PEPD) gene located at 19q12-q13.11, prolidase is a ubiquitous cytosolic enzyme that catalyzes hydrolysis of oligopeptides with a C terminal proline or hydroxyproline. We describe here four Amish children with a severe phenotype of prolidase deficiency in the Geauga settlements of Ohio as the first report of prolidase deficiency in the Amish population as well as in the United States. The patients presented with infection, hepatosplenomegaly, or thrombocytopenia, in contrast to most cases previously reported in the literature, presenting with skin ulcers. All four patients had typical facial features, classic skin ulcers, and multisystem involvement. Recurrent infections, asthma-like chronic reactive airway disease, hyperimmunoglobulins, hepatosplenomegaly with mildly elevated aspartate transaminase (AST), anemia, and thrombocytopenia were common and massive imidodipeptiduria was universal. Prolidase activity in our patients is nearly undetectable. Direct sequencing of PCR-amplified genomic DNA for all of the exons from the four patients revealed the same homozygous single nucleotide mutation c.793 T > C in exon 11, resulting in a premature stop-codon at amino acid residue 265 (p.R265X). It is speculated that the severe phenotype in these patients might be associated with the type of the PEPD gene mutation. PMID- 16470700 TI - A 9-year-old male with a duplication of chromosome 3p25.3p26.2: clinical report and gene expression analysis. AB - We describe a 9-year-old male referred for genetic evaluation for Prader-Willi syndrome (PWS). PWS is the most common genetically defined cause of life threatening obesity and results from a functional loss of paternally expressed genes from the chromosome 15q11-q13 region. The patient presented with pervasive developmental disorder, delayed speech, and rapid onset of obesity at age 4 years, all features similar to PWS. However, chromosome 15q11-q13 methylation testing and fragile X studies were normal. GTG-banding and fluorescence in situ hybridization (FISH) with whole chromosome 3 paint probe (WCP3) and a chromosome 3p subtelomeric probe suggested a duplication of 3p25.3p26.2, a finding supported by comparative genomic hybridization (CGH). This region of chromosome 3p contains genes which contribute to obesity and behavioral problems, most notably, ghrelin (GHRL), an oxytocin receptor (OXTR), solute carrier family six members (gamma aminobutyric acid (GABA) neurotransmitter transporters, SLC6A1 and SLC6A11), and peroxisome proliferator-activated receptor gamma (PPARG). To characterize these obesity and behavior related genes in our subject, we performed quantitative RT PCR and compared expression levels with similarly aged male subjects (four non obese males, four obese males, and four PWS males-two with 15q11-q13 deletions and two with maternal disomy 15). Our studies suggest increased expression of several genes in the 3p duplication region, including GHRL and PPARG, which may contribute to the phenotypic features in our 3p duplication subject. PMID- 16470702 TI - Twins with mental retardation and an interstitial deletion 7q34q36.2 leading to the diagnosis of long QT syndrome. PMID- 16470704 TI - Liquid chromatography/tandem mass spectrometry characterization of oxidized amyloid beta peptides as potential biomarkers of Alzheimer's disease. AB - Alzheimer's disease is characterized by the deposition of senile plaques that consist primarily of amyloid beta peptides. There is substantial evidence that amyloid beta is oxidized in vivo, which has led to the suggestion that oxidative stress is an important mediator of Alzheimer's disease. Metal-catalyzed oxidation can mimic in vivo oxidation of amyloid beta because the metal ion binds to the amino acid residues at the site of oxidation, which then deliver reactive oxygen species to that site. Based on electrospray mass spectrometry, it has been suggested that metal-catalyzed oxidation occurs on histidines-13 and -14. Unfortunately, the amyloid beta peptides provide complex spectra, so it is difficult to definitively characterize the sites of oxidation. Trypsin digestion of both native and oxidized amyloid beta1-16 and amyloid beta1-40 resulted in the formation of tryptic peptides corresponding to amyloid beta6-16, which could be separated by liquid chromatography (LC). Sites of oxidation were then unequivocally characterized as histidine-13 and histidine-14 by LC/tandem mass spectrometric (MS/MS) analysis of the tryptic peptides. The ability to analyze the specific amyloid beta6-16 tryptic fragments derived from full-length amyloid beta peptides will make it possible to determine whether oxidation in vivo occurs at specific histidine residues and/or at other amino acid residues such as methionine-35. Using methodology based on LC/MS/MS it will also be possible to analyze the relative amounts of oxidized peptides and native peptide in cerebrospinal fluid from patients with Alzheimer's disease as biomarkers of oxidative stress. PMID- 16470705 TI - Distinctive mass losses of tryptic peptides generated by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight. PMID- 16470706 TI - Mosaic trisomy 8 and Townes-Brocks syndrome due to a novel SALL1 mutation in the same patient. PMID- 16470707 TI - Who does the hat fit? Teenager heterogeneity and the effectiveness of information policies in preventing cannabis use and heavy drinking. AB - This paper models heterogeneity in the relationship between exposure to information at school or in the media and cannabis use and heavy drinking, using latent class techniques applied to data on French teenagers collected in 1993. Teenagers cluster in five classes which differ in their tastes for drunkenness and cannabis, and in the correlations between information exposure and cannabis use or heavy drinking. Teenager heterogeneity and habit-formation or precociousness effects limit the effectiveness of general information policies. Improving the impact of prevention requires that interventions be better targeted and personalised. We show how economic theory, latent class techniques and existing psychometric questionnaires can be used to build simple statistical tools for targeting prevention policies. PMID- 16470708 TI - Severe, fetal-onset form of olivopontocerebellar hypoplasia in three sibs: PCH type 5? AB - We present three siblings with a precise onset of fetal seizure-like activity who had severe olivopontocerebellar hypoplasia (OPCH) and degeneration. Autopsies at 20, 27, and 37 weeks gestation showed diffuse central nervous system volume loss that was most marked for the cerebellum and brain stem structures. Neuropathological abnormalities included dysplastic, C-shaped inferior olivary nuclei, absent or immature dentate nuclei, and cell paucity more marked for the cerebellar vermis than the hemispheres. Delayed development was seen in layer 2 of the cerebral cortex and in Purkinje cells of the cerebellum. Prenatal monitoring defined a developmental window of 16-18 weeks gestation when ultrasonic assessment of cerebellar width was used for prenatal diagnosis. We discuss our findings in the context of the differential diagnosis for infantile (O)PCH and propose a classification scheme for the pontocerebellar hypoplasias. These patients represent the earliest reported with OPCH and provide unique information regarding the developmental neuropathology of this condition. PMID- 16470709 TI - Screening for metabolically stable aryl-propionamide-derived selective androgen receptor modulators for doping control purposes. AB - Anabolic agents have been among the most frequently detected drugs in amateur and professional sport. A novel class of therapeutics presumably complementing anabolic steroids in the near future includes so-called selective androgen receptor modulators (SARMs) that have been under clinical investigations for several years. Although not yet commercially available, their potential for misuse in sports is high. Four aryl-propionamide-derived SARMs were synthesized in order to establish a fast and robust screening procedure using liquid chromatography/electrospray ionization tandem mass spectrometry. Synthesized compounds were characterized by high-resolution/high-accuracy mass analysis employing a linear ion trap-Orbitrap hybrid mass spectrometer while routine analyses were conducted on a triple-quadrupole mass spectrometer. Characteristic product ions obtained by collision-induced dissociation were found at m/z 289 and 261 as well as m/z 269 and 241 representing the bisubstituted aniline residues of selected model compounds. Assay validation was performed regarding lower limit of detection (1 ng/mL), recovery (85-105%), intraday precision (7.6-11.6%) and interday precision (9.9-14.4%), and precursor ion scan experiments on diagnostic product ions enabled the detection of a structurally related compound at 50 ng/mL. PMID- 16470710 TI - Mechanisms of change in mentalization-based treatment of BPD. AB - There are very few less contentious issues than the role of attachment in psychotherapy. Concepts such as the therapeutic alliance speak directly to the importance of activating the attachment system, normally in relation to the therapist in individual therapy and in relation to other family members in family based intervention, if therapeutic progress is to be made. In group therapy the attachment process may be activated by group membership. The past decade of neuroscientific research has helped us to understand some key processes that attachment entails at brain level. The article outlines this progress and links it to recent findings on the relationship between the neural systems underpinning attachment and other processes such as making of social judgments, theory of mind, and access to long-term memory. These findings allow intriguing speculations, which are currently undergoing empirical tests on the neural basis of individual differences in attachment as well as the nature of psychological disturbances associated with profound disturbances of the attachment system. In this article, we explore the crucial paradoxical brain state created by psychotherapy with powerful clinical implications for the maximization of therapeutic benefit from the talking cure. PMID- 16470711 TI - Interpersonal psychotherapy for borderline personality disorder: possible mechanisms of change. AB - This article describes the rationale for applying interpersonal psychotherapy (IPT) to the treatment of patients who have borderline personality disorder and explains the adaptation of standard IPT to that end. The authors describe the preliminary stages of adapting IPT for a pilot study to test its feasibility and potential efficacy and speculate on potentially therapeutic mechanisms for IPT in treating patients who have borderline personality disorder. PMID- 16470712 TI - Efficacy and safety of radiofrequency ablation for perivascular hepatocellular carcinoma without hepatic inflow occlusion. AB - BACKGROUND: The role of radiofrequency ablation (RFA) for perivascular (up to 5 mm from the major intrahepatic portal vein or hepatic vein branches) hepatocellular carcinoma (HCC) is unclear because of possible incomplete tumour ablation and potential vascular damage. This study aimed to evaluate the safety and efficacy of RFA for perivascular HCC without hepatic inflow occlusion. METHODS: Between May 2001 and November 2003, RFA using an internally cooled electrode was performed on 52 patients with perivascular HCC (group 1) through open (n = 39), percutaneous (n = 9), laparoscopic (n = 2) and thoracoscopic (n = 2) approaches. Hepatic inflow occlusion was not applied during the ablation procedure. The perioperative and postoperative outcomes were compared with those of 90 patients with non-perivascular HCC (group 2) treated by RFA during the same period. RESULTS: The morbidity rate was similar between groups 1 and 2 (25 versus 28 per cent; P = 0.844). One patient in group 1 (2 per cent) and two in group 2 (2 per cent) had developed thrombosis of major intrahepatic blood vessels on follow-up computed tomography scan. There were no significant differences between groups 1 and 2 in mortality rate (2 versus 0 per cent; P = 0.366), complete ablation rate for small HCC (92 versus 98 per cent; P = 0.197), local recurrence rate (11 versus 9 per cent; P = 0.762) and overall survival (1-year: 86 versus 87 per cent; 2-year: 75 versus 75 per cent; P = 0.741). CONCLUSION: RFA without hepatic inflow occlusion is a safe and effective treatment for perivascular HCC. PMID- 16470713 TI - Routine surgical pathology in general surgery. AB - BACKGROUND: Although pathological analysis provides the definitive diagnosis for most resection specimens, recent evidence suggests that such analysis may be omitted for certain routine samples. This was a retrospective analysis of the value of routine histopathological examination performed in daily general surgical practice. METHODS: All specimens from routine appendicectomies, cholecystectomies, haemorrhoidectomies and inguinal hernia repairs performed between 1993 and 2002 were included. The analysis included a comparison of histological and macroscopic diagnoses, review of preoperative and peroperative findings, and an evaluation of the consequences of routine histopathological assessment on patient management and costs. RESULTS: With the exception of hernia specimens, the rate of submission for routine pathological evaluation was 100 per cent. No hernia sac specimen from more than 2000 interventions revealed aberrant histological findings. Of 311 haemorrhoidectomy specimens three showed malignancy, all of which had a suspicious macroscopic appearance. Of 1465 appendices, only one (0.1 per cent) had a potentially relevant histological diagnosis that was not suspected macroscopically. Among 1523 cholecystectomy specimens, all adenomas (0.6 per cent) and carcinomas (0.4 per cent) were suspected macroscopically or developed in association with a known disease. CONCLUSION: The rarity of incidental histological findings relevant to patient management, especially in the absence of macroscopic abnormalities, suggests that routine histological examination of certain specimens may be omitted. A more elementary role for macroscopic examination of the specimen by the surgeon and the pathologist is proposed. PMID- 16470714 TI - Mechanisms of change in dialectical behavior therapy: theoretical and empirical observations. AB - Dialectical behavior therapy (DBT) can be considered a well-established treatment for borderline personality disorder (BPD) as evidenced by seven well-controlled randomized clinical trials across four independent research teams. The primary purpose of this article is to address a variety of potential mechanisms of change that may be associated with those aspects of DBT that are unique to the treatment and its theoretical underpinnings. Based on the biosocial theory of BPD, many of these mechanisms can be distilled down to the following process: the reduction of ineffective action tendencies linked with dysregulated emotions. Specifically we address the following interventions and associated mechanisms of change: mindfulness, validation, targeting and chain analysis, and dialectics. Patient change in BPD is conceptualized primarily as helping the patient to engage in functional, life-enhancing behavior, even when intense emotions are present. Ultimately, our goal was to provide guidance for theoretically and empirically grounded research on the mechanisms of change in DBT. PMID- 16470715 TI - Screening for amphetamine and amphetamine-type drugs in doping analysis by liquid chromatography/mass spectrometry. AB - A selective and sensitive method for the qualitative screening of urine samples for 27 amphetamine and amphetamine-type drugs in the field of doping analysis is described. The method consists of a liquid-liquid extraction with diethyl ether at pH 14 and analysis of the extracts with a LCQ-Deca mass spectrometer equipped with an atmospheric pressure chemical ionisation interface, operated in positive ionisation mode. The total run time was 15 min. All compounds were analysed in MS2 or MS3. The detection limit for all compounds was lower than 25 ng/mL except for chlorphentermine (detection limit: 250 ng/mL). PMID- 16470716 TI - Hypothesized mechanisms of change in cognitive therapy for borderline personality disorder. AB - Preliminary evidence suggests that cognitive therapy (CT) is effective in treating borderline personality disorder (BPD). According to cognitive theory, BPD patients are characterized by dysfunctional beliefs that are relatively enduring and inflexible and that lead to cognitive distortions such as dichotomous thinking. When these beliefs are activated, they lead to extreme emotional and behavioral reactions, which provide additional confirmation for the beliefs. It is hypothesized that a change in dysfunctional beliefs is the primary mechanism of change associated with CT. However, additional mechanisms of change are likely also at work in CT, including enhancement of skills, reduction in hopelessness, and improvement in attitude toward treatment. Each of these mechanisms is discussed in light of cognitive theory, data from an open clinical CT trial, relevant literature, and therapeutic interventions. Findings from the CT trial support the role of cognitive change during therapy and its continuation after termination. PMID- 16470717 TI - Economic evaluation of health effects with preference imprecision. AB - The estimation of economic benefits of health effects through direct methods, such as contingent valuation, presents the problem of preference imprecision. This paper deals with this problem by proposing an elicitation method that allows the subject to state an interval for willingness to pay, without inducing any specific amount as a response. The paper also analyzes the effects of the context where changes in health occur on the associated imprecision level and the estimates, by comparing a situation without context with another in which effects are due to atmospheric pollution. The econometric modelling develops a Bayesian estimation method for censored intervals, which models the existing uncertainty between the lower and upper limits derived from the elicitation process. Results prove that data dispersion is significantly higher for the non-contextual scenario, and increases for the most severe symptoms. PMID- 16470718 TI - Self-reported health: reliability and consequences for health inequality measurement. AB - Self-reported health (SRH) is one of the most frequently employed measures for assessing income-related health inequalities between counties. A previous study has shown that 28% of respondents changed their assessment of their health status when asked a SRH question on two occasions in the same survey (first as part of self-completed questionnaire and then in a personal interview). This study re examines this issue using another survey where SRH was again asked twice of respondents, but this time the personal interview was first and self-completion second. We find the same variation in responses, but the predominant direction is away from the 'extreme' categories 'Excellent' and 'Poor' which is the opposite direction to the previous study. We therefore conclude that the most likely explanation is a mode of administration effect that makes people less likely to choose the extreme categories in a self-completion questionnaire, but not a personal interview. However, this effect has a relatively minor impact on measures of inequality. This is due to a large proportion of the movement (i.e. movement to the middle) not being related to income and hence does not systematically impact on the cumulative distribution of health across this measure of socio-economic status. PMID- 16470719 TI - Characterization and functional investigation of single nucleotide polymorphisms (SNPs) in the human TLR5 gene. AB - Toll-like receptors recognize pathogen-associated molecular patterns (PAMPs) and TLR5 is the pathogen recognition receptor (PRR) for bacterial flagellin. Patients carrying a R392 stop polymorphism display an inflammatory phenotype and increased susceptibility to pneumonia caused by the flagellated bacteria Legionella pneumophila. While this suggests that TLR5 mutations may be clinically relevant, functional data are not available for the majority of the other TLR5 polymorphisms. We have characterized all known single nucleotide polymorphisms (SNPs) of TLR5 for their functional relevance upon stimulation in transiently transfected CHO-K1 cells. Among the 13 missense SNPs of TLR5 reported in the human genetic databases, three SNPs (c.1174C>T, p.R392X; c.2081A>G, p.D694G; and c.2464C>T, p.L822F) were found to be functionally relevant in transiently transfected CHO-K1 cells. The prevalences of these functionally relevant SNPs in our investigation were 11.9 %, 0 %, and 0 %, in healthy donors. The p.D694G and p.L822F SNPs are of low frequency in the Caucasian population though further investigations of the common p.R392X variant alone or of functional relevant TLR5 SNPs in combination with other TLR SNPs will elucidate their possible role on disease susceptibility in humans and may facilitate clinical diagnosis. PMID- 16470720 TI - Cell-type-specific limitation on in vivo serotonin storage following ectopic expression of the Drosophila serotonin transporter, dSERT. AB - The synaptic machinery for neurotransmitter storage is cell-type specific. Although most elements of biosynthesis and transport have been identified, it remains unclear whether additional factors may be required to maintain this specificity. The Drosophila serotonin transporter (dSERT) is normally expressed exclusively in serotonin (5-HT) neurons in the CNS. Here we examine the effects of ectopic transcriptional expression of dSERT in the Drosophila larval CNS. We find a surprising limitation on 5-HT storage following ectopic expression of dSERT and green fluorescence protein-tagged dSERT (GFP-dSERT). When dSERT transcription is driven ectopically in the CNS, 5-HT is detectable only in 5-HT, dopamine (DA), and a very limited number of additional neurons. Addition of exogenous 5-HT does not dramatically broaden neuronal storage sites, so this limitation is only partly due to restricted intercellular diffusion of 5-HT. Furthermore, this limitation is not due to gross mislocalization of dSERT, because cells lacking or containing 5-HT show similar levels and subcellular distribution of GFP-dSERT protein, nor is it due to lack of the vesicular transporter, dVMAT. These data suggest that a small number of neurons selectively express factor(s) required for 5-HT storage, and potentially for function of dSERT. PMID- 16470721 TI - A novel mutation in the GATA4 gene in patients with Tetralogy of Fallot. AB - In vertebrates, heart formation which integrates different structures and cell types is a complex process that involves a network of genes regulated by transcription factors. Proper spatiotemporal expression of these factors ensure the highly needed tight control of each step in organogenesis. A mistake at any step from cell-commitment to valve formation will have a major impact on heart morphogenesis and function leading to congenital heart disease (CHD). Cardiac abnormalities occur with an incidence of one per 100 live births and represent 25% of all congenital malformations. As an alternative approach to linkage analysis of familial cases of CHD, we started screening familial and sporadic cases of CHDs in a highly consanguineous population for mutations in genes encoding cardiac-enriched transcription factors. The evolutionarily conserved role of these proteins in cardiac development suggested a role in CHD. In this study, we report a mutation in the gene encoding GATA4, one of the earliest markers of heart development. This mutation was found in two out of 26 patients with Tetralogy of Fallot (TOF), and in none of the 94 patients with different phenotypes included in the study, nor in 223 healthy individuals. The heterozygous mutation results in an amino acid substitution in the first zinc finger of GATA4 that reduced its transcriptional activation of downstream target genes, without affecting GATA4 ability to bind DNA, nor its interaction with ZFPM2. PMID- 16470722 TI - Study of transaldolase deficiency in urine samples by capillary LC-MS/MS. AB - Transaldolase (TAL) is a key enzyme of the pentose phosphate pathway (PPP). TAL deficiency is a newly recognized cause of liver cirrhosis. We have developed an ion-pair LC separation combined with negative ion electrospray MS/MS detection method to assess PPP metabolites in urine samples from TAL-deficient mice. Sedoheptulose 7-phosphate (S7P), C5-polyols D-arabitol and D-ribitol, and 6 phosphogluconate (6PG) levels were markedly increased in urine of TAL-deficient mice with respect to those of wild-type and heterozygote littermates. The detection limits of S7P, D-arabitol, and 6PG were 0.15 +/- 0.015 pmol, 3.5 +/- 0.41 pmol, and 0.61 +/- 0.055 pmol, respectively. The limit of quantitation was 0.4 +/- 0.024 nmol/ml for S7P, 1.6 +/- 0.11 nmol/ml for 6PG and 10 +/- 0.7 nmol/ml for D-arabitol. Additional metabolites, hexose 6-phosphates (m/z 259), D ribose 5-phosphate and D-xylulose 5-phosphate (m/z 229), D-fructose 1,6 diphosphate (m/z 339), C6-polyols (m/z 181) and GSSG (m/z 611), that have been positively identified in mouse urine, showed similar levels in control and TAL deficient mice. PMID- 16470723 TI - Matrix effects on accurate mass measurements of low-molecular weight compounds using liquid chromatography-electrospray-quadrupole time-of-flight mass spectrometry. AB - Liquid chromatography (LC) with high-resolution mass spectrometry (HRMS) represents a powerful technique for the identification and/or confirmation of small molecules, i.e. drugs, metabolites or contaminants, in different matrices. However, reliability of analyte identification by HRMS is being challenged by the uncertainty that affects the exact mass measurement. This parameter, characterized by accuracy and precision, is influenced by sample matrix and interferent compounds so that questions about how to develop and validate reliable LC-HRMS-based methods are being raised. Experimental approaches for studying the effects of various key factors influencing mass accuracy on low molecular weight compounds (MW < 150 Da) when using a quadrupole-time-of-flight (QTOF) mass analyzer were described. Biogenic amines in human plasma were considered for the purpose and the effects of peak shape, ion abundance, resolution and data processing on accurate mass measurements of the analytes were evaluated. In addition, the influence of the matrix on the uncertainty associated with their identification and quantitation is discussed. A critical evaluation on the calculation of the limits of detection was carried out, considering the uncertainty associated with exact mass measurement of HRMS-based methods. The minimum concentration level of the analytes that was able to provide a statistical error lower than 5 ppm in terms of precision was 10 times higher than those calculated with S/N = 3, thus suggesting the importance of considering both components of exact mass measurement uncertainty in the evaluation of the limit of detection. PMID- 16470724 TI - Host-defence skin peptides of the Australian Streambank Froglet Crinia riparia: isolation and sequence determination by positive and negative ion electrospray mass spectrometry. AB - A combination of positive and negative ion electrospray mass spectrometry (ES-MS) together with automated Edman sequencing has been used to determine the amino acid sequences of the host-defence peptides from the skin glands of the froglet Crinia riparia. The peptides are called riparins. Of the eight peptides isolated, five are neuropeptides containing intramolecular disulfide linkages; e.g. the major peptide riparin 1.4 (FFLPPCAYKGTC-OH). Positive ion ES-MS identifies the five residues of riparin 1.4 outside the disulfide moiety, but provides no information on the sequence within the disulfide ring. In contrast, the negative ion dissociations of the [M-H]- ion of riparin 1.4 identify the --S-S-- link by loss of H2S2 from the [M-H]- ion, and also provide the sequence within the disulfide unit. Other peptides are riparin 2.1 [(IIEKLVNTALGLLSGL-NH2), a narrow spectrum antibiotic], signiferin 3.1 [(GIAEFLNYIKSKA-NH2), an nNOS inhibitor] and riparin 5.1 [IVSYPDDAGEHAHKMG-NH2], which shows no neuropeptide, antibiotic or nNOS activity. PMID- 16470725 TI - Variable contribution of the MTHFR C677T polymorphism to non-syndromic cleft lip and palate risk in China. AB - Non-syndromic cleft lip with or without cleft palate (nsCL/P) is one of the most common craniofacial malformations among newborn infants. It has been demonstrated that periconceptional folic acid supplementation may reduce the occurrence of offspring with clefts, particularly in the North China; however, the mechanism remains unknown. Our study of a thermolabile polymorphism (C677T) of methylenetetrahydrofolate reductase (MTHFR) gene in 170 Chinese case-parent triads revealed a moderate association between this MTHFR polymorphism and nsCL/P in a population from North China, but not in a population from South China. Moreover, the study revealed that the heterozygous parents in the North were about twice as likely to transmit the high-risk T allele to affected cases, as that observed in the South (OR = 2.24, 95% CI: 1.08-4.65). Thus, the MTHFR polymorphism is a significant risk factor for nsCL/P in this Northern Chinese population. Our study suggested possible genetic heterogeneity in the development of nsCL/P among Northern and Southern populations in China. PMID- 16470726 TI - Delineation of a 2.2 Mb microdeletion at 5q35 associated with microcephaly and congenital heart disease. AB - Fine mapping of chromosomal deletions and genotype-phenotype comparisons of clinically well-defined patients can be used to confirm or reveal loci and genes associated with human disorders. Eleven patients with cytogenetically visible deletions involving the terminal region of chromosome 5q have been described, but the extent of the deletion was determined only in one case. In this study we describe a 15-year-old boy with Ebstein anomaly, atrial septal defect (ASD), atrioventricular (AV) conduction defect, and microcephaly. He had an apparently balanced paracentric inversion of chromosome 5, with the karyotype 46, XY,inv(5)(q13q35) de novo. Further mapping of the chromosome breakpoints using fluorescence in situ hybridization (FISH) revealed a 2.2 Mb microdeletion at the 5q35 breakpoint, which spans 16 genes, including the cardiac homeobox transcription factor gene NKX2-5. The current data suggest that haploinsufficiency of NKX2-5 cause Ebstein anomaly and support previous results showing that NKX2-5 mutations cause ASD and AV conduction defect. Furthermore, we suggest presence of a new microcephaly locus within a 2.2 Mb region at 5q35.1 q35.2. PMID- 16470727 TI - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry with size-exclusion chromatographic fractionation for structural characterization of synthetic aliphatic copolyesters. AB - We report matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and off-line coupling of size-exclusion chromatography with MALDI-TOFMS analysis (SEC/MALDI-TOFMS) methods for the detailed characterization of poly[(R,S)-3-hydroxybutyrate-co-L-lactic acid], P[(R,S)-3HB co-LA], and poly[(R,S)-3-hydroxybutyrate-co-epsilon-caprolactone], P[(R,S)-3HB-co CL], copolymer samples which are expected to be used in special medical application as scaffolds for cartilage and soft tissue engineering. The novel copolyesters contained randomly distributed (R,S)-3-hydroxybutyrate structural units, were synthesized by transesterification of the corresponding homopolymers, i.e. atactic poly[(R,S)-3-hydroxybutyrate], a-PHB, and poly(L-Lactide) (PLLA) or poly(epsilon-caprolactone) (PCL), respectively. The MS methods used for the characterization of the resulting polydisperse copolyester samples were supported by classical methods (NMR, SEC). The structures of individual copolyester macromolecules, including end-group chemical structures, were established using initially MALDI-TOFMS and then SEC/MALDI-TOFMS. The compositions of the copolyesters were determined by two methods, namely based on 1H NMR and MALDI-TOF spectra. The two sets of values showed good agreement. The sequence distribution was determined using the signal intensities of individual copolyester macromolecules, which appeared in MALDI-TOF mass spectra. Furthermore, sequence analysis gave information about the degree of transesterification. The copolyesters synthesized, with only one exception, were demonstrated to be almost random, which implies that the ester-ester exchange was close to completion. PMID- 16470728 TI - Simultaneous determination of tanshinone IIA and its three hydroxylated metabolites by liquid chromatography/tandem mass spectrometry. AB - A rapid and sensitive method based on liquid chromatography/tandem mass spectrometry (LC/MS/MS) for the simultaneous determination of tanshinone IIA and its three hydroxylated metabolites, tanshinone IIB, hydroxytanshinone IIA and przewaquinone A, in a rat liver microsome was developed and fully validated. A single step of liquid-liquid extraction with ethyl acetate was utilized in this method. Chromatographic separation of the sample matrix from the analytes and the internal standard diazepam was performed using a Shim-pack VP-ODS analytical column. Detection was performed on a triple quadrupole tandem mass spectrometer equipped with an electrospray ionization source and operated in selected reaction monitoring (SRM) mode. The method was linear in the concentration range of 1-500 ng/mL for all analytes. The intra- and inter-day precisions (RSD %) were within 15% and deviations of the assay accuracies were within 15.0% for all analytes. The analytes proved to be stable during sample storage, preparation and analyses. This validated method was successfully applied to the enzyme kinetic study of tanshinone IIA in liver microsome. The elimination of tanshinone IIA and formation of tanshinone IIB and hydroxytanshinone IIA in the liver microsome all exhibited a sigmoidal kinetics profile. The formation of przewaquinone A shows a typical hyperbolic profile. In addition, this method has now been applied in the analysis of other bio-samples including plasma, urine, bile and feces. PMID- 16470729 TI - Evidence of genetic underexpression in chorionic villi samples of euploid fetuses with increased nuchal translucency at 10-11 weeks' gestation. AB - OBJECTIVE: To retrospectively investigate whether the genetic profile from chorionic villous sampling (CVS) found in euploid fetuses with increased NT differs from matched controls. STUDY DESIGN: We employed cDNA microarray technology to characterize and compare the gene expression profile of chorionic villous tissues (which encompass the trophoblast and inner mesenchymal core) belonging to four singleton male fetuses with increased NT at 10-11 weeks' gestation. A pool of four normal chorionic villous tissues belonging to four respective fetuses, matched for gestational age and gender, was used as controls. RESULTS: In euploid fetuses, we found several underexpressed genes, possibly involved in mechanisms associated with the abnormal NT thickness. All these genes are likely to belong to the mesenchymal core of the villus that originates from the extraembryonic mesoderm, and thus might be closely representative of the embryonic genetic profile. They include: (1) genes of embryonic development and differentiation such as Endothelin 3 (EDN3) and secreted frizzled-related protein 4 (SFRP4); (2) genes of the extracellular matrix (ECM) metabolism such as tissue inhibitor of metalloproteinase1 (TIMP1), and disintegrin-like and matrix metalloproteinase (MMP) (reprolysin type) with thrombospondin type 1 Motif or ADAMTS2, exostoses (multiple)-like 1 (EXTL1), heparan sulfate (HS) 6-O sulfotransferase 1 or HS6ST1, fibronectin 1 (FN1) and Integrin Alpha 10 (ITGA10) involved in HS and proteoglycan bio-synthesis, ECM synthesis and cell-matrix adhesion; (3) genes involved in vessel formation and differentiation such as angiogenic factor (VG5Q), and in blood pressure control and muscle contraction, like Endothelin 3 or EDN3 and sarcolemma associated protein (SLMAP). Such lower expressions of the villous tissues might be related to an immature genetic profile of the embryo development as well as abnormal regulation of ECM bio synthesis and/or improper vessel growth and blood pressure control. Also, the results partially support the theories proposed for NT enlargement such as altered composition of ECM and abnormal/delayed development of the circulatory system. CONCLUSIONS: Abnormal extraembryonic genetic expression is found at 10-11 weeks' gestation in euploid fetuses with increased NT. If both extra- and intraembryonic mesoderms express the same genetic alterations, then microarray analyses on CVS could be used to screen several mesoderm-derivate anomalies. PMID- 16470730 TI - Clinical findings in 33 subjects with large supernumerary marker(15) chromosomes and 3 subjects with triplication of 15q11-q13. AB - We present clinical data on 33 subjects with additional copies of the Prader Willi-Angelman critical region (PWACR) contained in a supernumerary marker chromosome (SMC). Twenty-three subjects had a typical large non-mosaic SMC(15) containing two copies of the PWACR. They showed a variable but generally severe phenotype of learning disability and autism, with seizures in approximately two thirds. The other 10 differed from this typical pattern in respect of mosaicism, variation in copy number, or arrangement of the PWACR within the SMC or number of SMC per cell. Clinical severity increased with the number of additional copies of the PWACR and decreased with mosaicism for a normal cell line. There was a trend for a larger number of seizures to be associated with more severe learning disability. Three subjects with interstitial triplications of 15q11-q13 showed a range of phenotypes similar to those of the typical large SMC(15). All additional copies of the PWACR in this series were maternally-derived. FISH and molecular data localizing the breakpoints of the rearrangements have been previously published or are included in this report. No correlations were found between specific clinical features and variations in breakpoints proximal and distal to the PWACR. PMID- 16470731 TI - Dyggve-Melchior-Clausen syndrome and Smith-McCort dysplasia: clinical and molecular findings in three families supporting genetic heterogeneity in Smith McCort dysplasia. AB - Dyggve-Melchior-Clausen syndrome (DMC) (MIM 223800) and Smith-McCort dysplasia (SMC) (MIM 607326) are rare allelic autosomal recessive spondylo-epi-metaphyseal dysplasias (SEMDs) characterized by similar skeletal manifestations. Both phenotypes have been mapped to chromosome 18q21.1 and mutations in the DYM (dymeclin) gene were identified in 13 families with DMC and in two families with SMC. Most mutations identified in DMC predict a loss of function, while those identified in SMC are mainly missense mutations, presumably associated with residual DYM activity and a less severe phenotype. We studied three consanguineous families from Turkey, Lebanon, and Georgia, one with SMC and two with DMC and identified different homozygous DYM mutations (IVS3 194-1G > A, 938_942delTGTCT) in the DMC families. No mutation was identified in the SMC family, possibly suggesting genetic heterogeneity of this disorder. PMID- 16470733 TI - Inverse association between severe nausea and vomiting in pregnancy and some congenital abnormalities. AB - The objective of the study was to investigate the possible association between nausea and vomiting in early pregnancy and congenital abnormalities. The prevalence of medically-recorded severe nausea and vomiting in early pregnancy in cases with congenital abnormalities and their available matched population controls without any defect was compared in the population-based large data set of the Hungarian Case-Control Surveillance System of congenital abnormalities, 1980-1996. Of 22,843 cases with as 25 different congenital abnormality groups, 1,713 (7.5%) cases had mothers with medically recorded and treated severe nausea and vomiting during pregnancy. Of 38,151 matched population controls, 3,777 (9.9%) had mothers with severe nausea and vomiting (adjusted prevalence odds ratio (POR) with 95% CI: 0.74, 0.68-0.79). Five congenital abnormality groups: cleft lip with or without cleft palate (0.50, 0.37-0.70), posterior cleft palate (0.53, 0.32-0.89), renal a/dysgenesis (0.23, 0.06-0.96), obstructive defects of urinary tract (0.32, 0.18-0.58), and cardiovascular malformations (0.68, 0.57 0.81) had mothers with a lower prevalence of severe nausea and vomiting in pregnancy (adjusted PORs with 95% CI included in parentheses). Of 25 congenital abnormality groups, 22 had POR lower than 1. Thus in this study the mothers of cases with congenital abnormalities were 26% less likely to have had severe nausea and vomiting in early pregnancy than the mothers of population controls without congenital abnormalities. PMID- 16470734 TI - Prenatal diagnosis of de novo t(2;18;14)(q33.1;q12.2;q31.2), dup(5)(q34q34), del(7)(p21.1p21.1), and del(10)(q25.3q25.3) and a review of the prenatally ascertained de novo apparently balanced complex and multiple chromosomal rearrangements. AB - OBJECTIVES: To present the prenatal diagnosis of a de novo complex chromosomal rearrangement (CCR) associated with de novo interstitial deletions and duplication and to review the literature. CASE AND METHODS: Amniocentesis was performed at 18 weeks' gestation because of an increased risk for Down syndrome based on maternal serum alpha-fetoprotein and human chorionic gonadotrophin screening. Amniocentesis revealed a karyotype of 46,XY,t(2;18;14)(q33.1;q12.2;q31.2),dup(5)(q34q34),del(7)(p21.1p21.1), del(10)(q25.3q25.3). The parental karyotypes were normal. The pregnancy was terminated. The fetus manifested facial dysmorphism, clinodactyly of both hands, and hypoplasia of the left great toe. Spectral karyotyping (SKY), cytogenetic polymorphism, and polymorphic DNA markers were used to investigate the imbalances and the origin of the de novo aberrant chromosomes. RESULTS: SKY showed a three way CCR. Cytogenetic polymorphism investigation of the derivative chromosome 14 of the fetus and the parental chromosomes 14 determined the maternal origin of the translocation. Polymorphic DNA marker analysis confirmed the maternal origin of the de novo interstitial deletions and duplication. No cryptic imbalance at or near the breakpoints of the CCR was detected by the molecular analysis. CONCLUSIONS: De novo apparently balanced CCRs may be associated with imbalances in other chromosomes. We suggest further investigation and re-evaluation of cryptic or subtle imbalances in all cases classified as de novo apparently balanced CCRs. PMID- 16470732 TI - Minimal phenotype in a girl with trisomy 15q due to t(X;15)(q22.3;q11.2) translocation. AB - Few cases of de novo unbalanced X;autosome translocations associated with a normal or mild dysmorphic phenotype have been described. We report a 3-year-old dizygotic female twin with prenatally ascertained increased nuchal translucency. Prenatal chromosome studies revealed nearly complete trisomy 15 due to a de novo unbalanced translocation t(X;15)(q22;q11.2) confirmed postnatally. A mild phenotype was observed with normal birth measurements, minor facial dysmorphic features (hypertelorism, short broad nose, and a relatively long philtrum), and moderate developmental delay at the age of 3 years in comparison to her male fraternal twin. Replication timing utilizing BrdU and acridine-orange staining showed that the der(X) chromosome was late-replicating with variable spreading of inactivation into the translocated 15q segment. The der(X) was determined to be of paternal origin by analyses of polymorphic markers and CGG-repeat at FMR1. Methylation analysis at the SNRPN locus and analysis of microsatellites on 15q revealed paternal isodisomy with double dosage for all markers and the unmethylated SNRPN gene. The Xq breakpoint was mapped within two overlapping BAC clones RP11-575K24 and RP13-483F6 at Xq22.3 and the 15q breakpoint to 15q11.2, within overlapping clones RP11-509A17 and RP11-382A4 that are all significantly enriched for LINE-1 elements (36.6%, 43.0%, 26.6%, 22.0%, respectively). We speculate that the attenuated phenotype may be due to inactivation spreading into 15q, potentially facilitated by the enrichment of LINE-1 elements at the breakpoints. In silico analysis of breakpoint regions revealed the presence of highly identical low-copy repeats (LCRs) at both breakpoints, potentially involved in generating the translocation. PMID- 16470735 TI - Postmortem evaluation of 220 prenatally diagnosed fetuses with neural tube defects: detection of associated anomalies in a Turkish population. AB - OBJECTIVES: The aim of this study is to represent the distribution of disorders resulting from neural tube defects (NTDs). MATERIALS AND METHODS: This study was conducted on 220 prenatally diagnosed cases with NTDs. Fetuses were evaluated by physical examination, anthropometric measurements, X-rays, and photographs after termination of pregnancy. Chromosome analysis and autopsy were performed for 37 fetuses (16.8%) with additional malformations. RESULTS: In 29 out of 37 fetuses (78.4%), additional malformations were detected by prenatal ultrasonography, whereas in eight cases postmortem evaluation produced additional findings that were not detected prenatally. Fourteen of 37 (37.8%) and 65 of 220 (29.5%) fetuses had clubfoot, which was mostly secondary to NTDs. There was no difference in sex distribution between isolated NTDs and the group with additional abnormalities and among the groups anencephaly and anencephaly + anomaly, encephalocele and encephalocele + anomaly, spina bifida and spina bifida + anomaly. There was only one case, a female fetus, with iniencephaly in this group. Anencephaly was more frequent in cases with isolated NTDs (48.1%) than in those with additional anomalies (27%). There was no difference for other groups of NTDs. The most frequent disorder was vertebral segmentation defects, which were detected in 11 out of 37 cases (29.7%). CONCLUSIONS: Evaluation of associated malformations and confirmation of ultrasound findings can be performed by postmortem examination and simple X-ray studies for exact diagnosis, which strongly affects decisions on further pregnancies as well as genetic counseling. This method is straightforward, inexpensive and effective. PMID- 16470736 TI - The fetal mandible: an in utero sonographic evaluation between 11 and 31 weeks' gestation. AB - OBJECTIVE: To construct a growth chart of the mandible throughout gestation. METHODS: A prospective cross-sectional study of normal singleton pregnancies was conducted. Measurements of the fetal mandibular transverse and antero-posterior diameters were performed with high-resolution transvaginal and transabdominal ultrasonography in 490 pregnant women with singleton low-risk pregnancies between 11 and 31 weeks' gestation. RESULTS: The mandibular transverse and antero posterior diameters were recorded by week of gestation and the ratio was calculated: mandibular ratio (MR) = 1.7759 - 0.01047 x gestational week. There was a negative linear correlation (-1.047%) for each incoming week of gestation. Normal values (+/-1SD and 2SD) were established. CONCLUSION: The present data provides a normal range of fetal mandibular diameters during normal pregnancies and introduces a new parameter, the mandibular ratio, for the intrauterine assessment of the fetal mandibular development. PMID- 16470737 TI - Detection of maternal deoxyribonucleic acid in peripheral blood of premature and mature newborn infants. AB - BACKGROUND: Over the past decade, a lot of attention has been directed towards the fetomaternal and maternofetal transfer of nucleated cells and plasma DNA. In some autoimmune diseases, the fetal DNA is suspected to play an important role in the etiology of the disease. In the same way, the presence of maternal cells and free plasma DNA in fetal/newborn circulation gives rise to interesting questions. The aim of our study was to detect maternal deoxyribonucleic acid in the peripheral blood of premature and mature newborn infants. METHODS: In the case of eight RhD-positive mothers-RhD-negative newborn pairs, peripheral blood samples were collected from the newborn infants within 35-120 min after birth. The maternal origin DNA was determined by real-time PCR amplification of the exon 7 of the RhD-positive allele. RESULT: In all eight cases, the RhD exon 7 was amplified during the PCR reaction. CONCLUSION: The result of our study demonstrates that maternal DNA is present in the fetal peripheral circulation. The presence of maternally derived cells/DNA in the blood of newborn infants might have a role in the immunization of the newborn infants and also could be a possible explanation for 'grandmother effect' in the case of Rh-negative nulligravida patients. PMID- 16470738 TI - Sexual differentiation of identified motor terminals in Drosophila larvae. AB - In Drosophila, we have found that some of the motor terminals in wandering third instar larvae are sexually differentiated. In three out of the four body-wall muscle fibers that we examined, we found female terminals that produced a larger synaptic response than their male counterparts. The single motor terminal that innervates muscle fiber 5 produces an EPSP that is 69% larger in females than in males. This is due to greater release of transmitter from female than male synaptic terminals because the amplitude of spontaneous miniature EPSPs was similar in male and female muscle fibers. This sexual difference exists throughout the third-instar: it is seen in both early (foraging) and late (wandering) third-instar larvae. The sexual differentiation appears to be neuron specific and not muscle specific because the same axon produces Is terminals on muscle fibers 2 and 4, and both terminals produce larger EPSCs in females than males. Whereas, the Ib terminals innervating muscle fibers 2 and 4 are not sexually differentiated. The differences in transmitter release are not due to differences in the size of the motor terminals. For the terminal on muscle fiber 5 and the Is terminal on muscle fiber 4, there were no differences in terminal length, the number of branches, or the number of synaptic boutons in males compared to females. These sexual differences in neuromuscular synaptic physiology may be related to male-female differences in locomotion. PMID- 16470739 TI - Serotonin and cAMP induce excitatory modulation of a serotonergic neuron. AB - Serotonin (5-HT) is an excitatory neurotransmitter and neuromodulator. In the Aplysia nervous system it increases excitability and induces spike broadening in sensory neurons. It is released at the synaptic terminals of the metacerebral cells (MCCs) and modulates the feeding neural circuit and buccal muscles during the aroused feeding state. We report that MCC itself is depolarized by 5-HT and becomes excitable. 5-HT induces tonic spike activity and even spike-burst activity. Conceivably, this sensitivity to its own transmitter could provide positive feedback excitation of MCC. Voltage clamp analysis of isolated cultured MCCs shows that 5-HT reduces a calcium-dependent outward current at the resting potential (-60 mV), and enhances steady state inward currents between -55 and -30 mV and between -75 and -100 mV. 8-Br-cAMP has similar effects, suggesting that cAMP mediates the 5-HT effects, in part. A transient calcium current is enhanced at voltages more positive than -40 mV. Barium and cesium selectively block the 5 HT-induced inward current between -75 and -100 mV. Substitution of N-methyl-D glucamine for sodium and adding cobalt block this current, also indicating that it is a hyperpolarization-activated cation current. The 5-HT-induced inward current between -55 and -30 mV is also blocked by sodium substitution and added cobalt, suggesting that 5-HT increases a depolarization-activated cation current. The outward current that remains when sodium and calcium currents are blocked is reduced by 5-HT. Thus, 5-HT enhances two different cation currents and reduces potassium currents. PMID- 16470740 TI - Functional splicing assay shows a pathogenic intronic mutation in neurofibromatosis type 1 (NF1) due to intronic sequence exonization. AB - Genomic variations with no apparent effect ("neutral polymorphisms") may have a significant effect on splicing. The effect of this type of mutation is difficult to spot, unless a functional assay is undertaken. In our study, DNA sequencing of a patient with clinically defined neurofibromatosis type 1 (NF1) showed only a single polymorphism in intron 30 due to an A>G transition 279 nucleotides from the 3' splice site. Using a minigene splicing assay we conclusively show that this change produces a cryptic exon with a 3' SS defined by the nucleotide change and the unexpected activation of a very weak 5'SS. Further site directed mutagenesis studies aimed at identifying the signals involved in the cryptic exon inclusion were carried out. Interestingly we find that particular characteristics of the cryptic 5' SS are essential for its inclusion. Significantly an additional single nucleotide change disrupting the cryptic 5'ss consensus sequence rescues the effect of the pathogenetic mutation resulting in normal splicing. PMID- 16470741 TI - Wiedemann-Rautenstrauch syndrome's fibroblasts display a normal in vitro lifespan. PMID- 16470742 TI - Molecular cytogenetic analysis of a familial interstitial deletion Xp22.2-22.3 with a highly variable phenotype in female carriers. AB - We describe a familial interstitial deletion of 7.7-Mb involving Xp22.2-22.3. The deletion was transmitted from an asymptomatic mother to her two children with severe developmental delay, no speech development and autistic behavior. Assessment of the deletion boundaries by FISH and PCR analyses indicated that the deletions encompasses 27 genes. Several of these genes are associated with known disorders, like KAL1 (Kallmann syndrome), steroid sulfatase (STS) (X-linked ichtyosis), and arylsulfatase E (ARSE) (chondrodysplasia punctata). The deletion also includes all four VCX genes (VCX-A, VCX-B1, VCX-B, and VCX-C) and the neuroligin 4 (NLGN4) gene. VCX-A deficiency has been shown previously to be associated with mental retardation and NLGN4 mutations lead to mental retardation in conjunction with autism. Functional deficiency of both MRX genes, VCX-A and NLGN4, are most likely associated with the impaired cognitive development of the patients described here. The phenotype associated with the Xp deletion was highly variable in female carriers and might be attributed to unfavorable X inactivation. However, all the 27 genes included in the deleted interval escape X inactivation and are expressed at variable levels from the normal X chromosome. Thus, the overall X inactivation pattern and inter-individual expression variability of the genes in distal Xp might be determinants of the phenotype associated with the deletion. PMID- 16470743 TI - Hypoparathyroidism-retardation-dysmorphism syndrome in a girl: A new variant not caused by a TBCE mutation--clinical report and review. AB - Hypoparathyroidism-retardation-dysmorphism (HRD) or Sanjad-Sakati syndrome (SSS) (OMIM 241410) is a rare autosomal recessive (AR) inherited condition, characterized by congenital hypoparathyroidism (hypoPTH), retardation, seizures, and a typical facial dysmorphism, consisting of prominent forehead, deep-set eyes, and abnormal external ears. This disorder has been mapped to the long arm of chromosome 1 (1q42-q43) and mutations in the gene coding for tubulin-specific chaperone E (TBCE) have been identified as the cause of the disease. Mutations in the same gene were also reported in patients with AR Kenny-Caffey syndrome (KCS). We report on a 41/2-year-old girl with congenital hypoPTH, seizures, developmental delay, and a facial dysmorphism, compatible with HRD syndrome. Mutation analyses revealed no mutations in the TBCE gene. In addition, normal TBCE protein and alpha-tubulin immunostaining were observed in a lymphoblastoid line derived from the patient, excluding the TBCE gene as the causative gene of the syndrome in this patient. A de novo microduplication of probe RP11-262I1 on 4q35 in the proposita was detected by microarray analyses, but this could not be confirmed by additional studies. We review and discuss the clinical findings of our case and those of the other reported cases with SSS and AR KCS. We conclude that a second gene locus for this disorder seems probable and that 4q35 needs further evaluation as a candidate region. PMID- 16470744 TI - Lymphatic drainage patterns of head and neck cutaneous melanoma observed on lymphoscintigraphy and sentinel lymph node biopsy. AB - BACKGROUND: The purpose of this study was to evaluate lymphatic drainage patterns of head and neck cutaneous melanoma observed on preoperative lymphoscintigraphy and sentinel lymph node biopsy (SLNB) and determine discordancy from clinically predicted lymphatic drainage patterns. METHODS: We conducted a retrospective chart review of 114 patients with head and neck cutaneous melanomas evaluated with preoperative lymphoscintigraphy and SLNB from January 2001 through July 2004. RESULTS: At least one sentinel lymph node (SLN) was identified in 97% of cases. On preoperative lymphoscintigraphy, an SLN was identified in an area not clinically predicted in 49 cases (43%). The most common sites of discordancy were in areas not typically dissected in standard neck dissections, such as the postauricular region, or in areas of more distant drainage than described previously, such as the inferior or posterior neck. Their percentages of discordant cases were 51%, 27%, and 22%, respectively. The sites of regional recurrence occurred in two cases not predicted on preoperative lymphoscintigraphy and in two cases of failed SLNB. CONCLUSIONS: On the basis of preoperative lymphoscintigraphy and the results of SLNB, head and neck cutaneous melanomas do have expected lymphatic drainage patterns despite perceived discordancy with previously clinically predicted drainage patterns that are based on standard neck dissection specimens. These "discordant" sites can still harbor melanoma, and all sites predicted on preoperative lymphoscintigraphy still need to be explored. The four cases of recurrences underscore the importance of close follow-up for all patients regardless of the SLNB result. PMID- 16470746 TI - Discordant encephalocele in monozygotic twins. PMID- 16470745 TI - Phase II trial of taxol in salivary gland malignancies (E1394): a trial of the Eastern Cooperative Oncology Group. AB - BACKGROUND: Malignant tumors of the salivary glands make up approximately 5% of head and neck cancers. The Eastern Cooperative Oncology Group (ECOG) initiated a phase II evaluation of paclitaxel in patients with locally recurrent or metastatic salivary gland malignancies. METHODS: Chemo-naive patients with histologically confirmed recurrent or metastatic carcinoma of salivary gland origin (mucoepidermoid, adenocarcinoma, or adenoid cystic) were eligible. Patients were treated with paclitaxel, 200 mg/m(2) IV, every 21 days for a minimum of four cycles. RESULTS: Forty-five patients were treated. Eight partial responses were seen among the 31 patients with mucoepidermoid or adenocarcinoma histologic findings for a response rate of 26%. No responses were seen in the adenoid cystic carcinoma group. No significant difference in overall survival was found among these three histologic subgroups. CONCLUSION: Paclitaxel demonstrates moderate activity in salivary gland tumors of mucoepidermoid and adenocarcinoma histology. The poor response rate in adenoid cystic carcinoma is consistent with prior reports in this chemoresistant histologic subtype. PMID- 16470747 TI - Angelman syndrome 2005: updated consensus for diagnostic criteria. AB - In 1995, a consensus statement was published for the purpose of summarizing the salient clinical features of Angelman syndrome (AS) to assist the clinician in making a timely and accurate diagnosis. Considering the scientific advances made in the last 10 years, it is necessary now to review the validity of the original consensus criteria. As in the original consensus project, the methodology used for this review was to convene a group of scientists and clinicians, with experience in AS, to develop a concise consensus statement, supported by scientific publications where appropriate. It is hoped that this revised consensus document will facilitate further clinical study of individuals with proven AS, and assist in the evaluation of those who appear to have clinical features of AS but have normal laboratory diagnostic testing. PMID- 16470749 TI - The reactivity-selectivity principle: an imperishable myth in organic chemistry. AB - The reactivity-selectivity principle (RSP), once a tenet of organic chemistry, eroded during the 1970s and was more or less abandoned by 1980. Although it has been clear for more than 25 years that a decrease in selectivity with increasing reactivity can only be expected with certainty if diffusion control is approached, the RSP has survived as an intuitively appealing rule. This Minireview shows why selectivity cannot generally decrease with increasing reactivity and highlights the weaknesses of the theoretical foundations of the RSP. PMID- 16470748 TI - Study of four genes belonging to the folate pathway: transcobalamin 2 is involved in the onset of non-syndromic cleft lip with or without cleft palate. AB - Cleft lip with or without cleft palate (CL/P) is the most common inborn craniofacial anomaly. Affected individuals require extensive medical and psychosocial support. Although CL/P has a complex and poorly understood etiology, increasing evidence of folate pathway involvement has been collected. So far, only the MTHFR gene has been extensively investigated as a risk factor for CL/P, while little has been done to test genetic variations in the folate biosynthetic pathways that may influence the infant's susceptibility to these birth defects. To date, this paper presents the first attempt to verify the involvement of four genes belonging to the folate pathway in nonsyndromic cleft onset. We used a case parent triad design to test for linkage disequilibrium in the case of seven SNPs mapping on four different genes: transcobalamin 1 and 2 (TCN1 and TCN2), methionine synthase (MTR), and MTR reductase (MTRR). Our finding suggests that TCN2 is involved in causing CL/P. Indeed, significant overtransmission of the C allele was observed at the polymorphism c.776C>G (p.Pro259Arg) to the affected offspring (P=0.01). Results obtained with additional TCN2 polymorphisms suggest that c.776C>G may be functionally related to CL/P. However, because conflicting data exist with regard to the effect of the polymorphism in transcobalamin 2 function or in perturbing plasma levels of key molecules in the folate pathway, further investigation is warranted to confirm our data. PMID- 16470750 TI - Bidentate coordination of pyrazolate in low-coordinate iron(II) and nickel(II) complexes. PMID- 16470751 TI - Rhodium(I) complexes of a PBP ambiphilic ligand: evidence for a metal-->borane interaction. PMID- 16470752 TI - Mixed diketonate thiolate copper(I) precursors for materials synthesis: control of Cu2S-forming thermolysis pathways by manipulating Lewis acid and base cluster building blocks. PMID- 16470753 TI - Asymmetric sequential allylic transfer reaction for the synthesis of 2-(1 stannylvinyl)-1,3-diols: concise synthesis of (-)-avenaciolide and (-) isoavenaciolide. PMID- 16470754 TI - An unsaturated alpha,omega-dianionic oligosilane. PMID- 16470755 TI - Determination of acid-base dissociation constants of amino- and guanidinopurine nucleotide analogs and related compounds by capillary zone electrophoresis. AB - CZE has been applied for determination of acid-base dissociation constants (pKa) of ionogenic groups of newly synthesized amino- and (amino)guanidinopurine nucleotide analogs, such as acyclic nucleoside phosphonate, acyclic nucleoside phosphonate diesters and other related compounds. These compounds bear characteristic pharmacophores contained in various important biologically active substances, such as cytostatics and antivirals. The pKa values of ionogenic groups of the above compounds were determined by nonlinear regression analysis of the experimentally measured pH dependence of their effective electrophoretic mobilities. The effective mobilities were measured by CZE performed in series of BGEs in a broad pH range (3.50-11.25), at constant ionic strength (25 mM) and temperature (25 degrees C). pKa values were determined for the protonated guanidinyl group in (amino)guanidino 9-alkylpurines and in (amino)guanidinopurine nucleotide analogs, such as acyclic nucleoside phosphonates and acyclic nucleoside phosphonate diesters, for phosphonic acid to the second dissociation degree (-2) in acyclic nucleoside phosphonates of amino and (amino)guanidino 9 alkylpurines, and for protonated nitrogen in position 1 (N1) of purine moiety in acyclic nucleoside phosphonates of amino 9-alkylpurines. Thermodynamic pKa of protonated guanidinyl group was estimated to be in the range of 7.75-10.32, pKa of phosphonic acid to the second dissociation degree achieved values of 6.64 7.46, and pKa of protonated nitrogen in position 1 of purine was in the range of 4.13-4.89, depending on the structure of the analyzed compounds. PMID- 16470756 TI - Monolithic silica capillary column with coated cellulose tris(3,5 dimethylphenylcarbamate) for capillary electrochromatographic separation of enantiomers. AB - Monolithic silica capillary columns were prepared by a sol-gel process in fused silica capillaries with an inner diameter of 50 microm and were modified by coating of cellulose tris(3,5-dimethylphenylcarbamate). Influences of the factors in the modification process on enantiomer separations were investigated. The prepared columns were used to perform enantiomer separations by CEC. Fifteen and two pairs of enantiomers were separated under aqueous and nonaqueous mobile phases, respectively, and most of them were baseline-separated with very high column efficiencies. The Van Deemter curve was found flat under high linear velocity of the mobile phase, which indicated favorable kinetic properties of the prepared columns. Baseline separation of a pair of enantiomers was achieved in 90 s with high-column efficiency by short-end separation under high voltage. PMID- 16470757 TI - Highlighting the role of the chlorine substituents in the glycopeptide antibiotic balhimycin for chiral recognition in capillary electrophoresis. AB - The vancomycin-type glycopeptide antibiotic balhimycin (I) and its dehaloanalogue dechlorobalhimycin (III), which is characterized by the total substitution of the two chlorine atoms of I by hydrogen, were employed as chiral selectors for the enantioresolution of 11 racemic dansyl amino acids and six 2-arylpropionic acid nonsteroidal anti-inflammatory racemic drugs by CE. The observed enantioresolution capability of I for all test analytes is clearly higher than that observed for III. This result suggests that chlorine substituents of I played a major role in the enantioresolution of these test analytes. A dimerization-based mechanism is proposed in order to explain this phenomenon. The two chlorine substituents of each monomer, which mutually penetrate into the cavity of the adjacent molecule of the dimer, are assumed to promote dimerization and as a consequence also enantioresolution. PMID- 16470758 TI - Less common applications of monoliths: I. Microscale protein mapping with proteolytic enzymes immobilized on monolithic supports. AB - This review summarizes the recent contributions to the rapidly growing area of immobilized enzymes employing both silica and synthetic polymer-based monoliths as supports. Focus is mainly on immobilized proteolytic enzyme reactors designed for studies in proteomics. Porous monoliths emerged first as a new class of stationary phases for HPLC in the early 1990s. Soon thereafter, they were also used as supports for immobilization of proteins and preparation of both stationary phases for bioaffinity chromatography and enzymatic reactors. Organic polymer-based monoliths are typically prepared using a simple molding process carried out within the confines of a "mold" such as chromatographic column or capillary. Polymerization of a mixture comprising monomers, initiator, and porogenic solvent affords macroporous materials. In contrast, silica-based monoliths are first formed as a rigid rod from tetraalkoxysilane in the presence of PEG and subsequently encased with a plastic tube. Both types of monolith feature large through-pores that enable a rapid flow-through. Since all the solutions must flow through the monolith, the convection considerably accelerates mass transfer within the monolith. As a result, reactors including enzyme immobilized on monolithic support exhibit much higher activity compared to the reactions in solution. PMID- 16470759 TI - Regiospecific ligand oxygenation in iron complexes of a carboxylate-containing ligand mediated by a proposed Fe(v)-oxo species. PMID- 16470760 TI - Molecular crowding induces telomere G-quadruplex formation under salt-deficient conditions and enhances its competition with duplex formation. PMID- 16470761 TI - Enantioselective enzymatic reactions in miniemulsions as efficient "nanoreactors". PMID- 16470762 TI - The transport of nanosized gene carriers unraveled by live-cell imaging. PMID- 16470763 TI - Photoswitching of the dielectric constant of the spin-crossover complex [Fe(L)(CN)2]H2O. PMID- 16470764 TI - Chloroplastic glycolipids fuel aldehyde biosynthesis in the marine diatom Thalassiosira rotula. AB - Enzymatic preparations and specialized analytical tools have shown that chloroplast-derived glycolipids are the main substrates for the biosynthetic pathway that produces antiproliferative polyunsaturated aldehydes in broken cells of the marine diatom Thalassiosira rotula. This process, which is associated with the formation of free fatty acids and lyso compounds from polar lipids but not triglycerides, is largely dependent on glycolipid hydrolytic activity, rather than phospholipase A(2) as previously suggested. Preliminary characterization of lipolytic enzymes has revealed protein bands of 40-45 kDa. Under native conditions these proteins seem to be associated with soluble aggregates that have an apparent molecular weight of approximately 200 kDa. The biochemical process, which is similar to that described in the algal-bloom forming diatom Skeletonema costatum, suggests a mechanism based on decompartmentalization and mixing of preexisting enzymes and substrates. PMID- 16470765 TI - Aminopeptidase p mediated detoxification of organophosphonate analogues of sarin: mechanistic and stereochemical study at the phosphorus atom of the substrate. AB - The activity of the aminopeptidase P from Escherichia coli in hydrolyzing a series of organophosphonate sarin analogues (1-6) was evaluated. The enzymatic rates of hydrolysis for methylphosphonate 1 with a methoxy group attached to the phosphorus center were 7- to 15-fold higher than those for the corresponding analogues 2-6. Double mutant R153W/R370L was able to hydrolyze the S(p) enantiomer of racemic 1 at a considerable rate. This mutant allowed the preparation of the R(p) isomer of the sarin analogue 1. All the mutants, R370L, R153A, W88L, R153L/R370L, and R153W/R370L, preferred the formation of (S(p))-8 to that of the corresponding (R(p))-8 enantiomer and displayed a better enantiomeric excess of products, by 1.4- to 2-fold as compared to the wild-type enzyme. Enzymatic hydrolysis of O,O-diisopropyl-p-nitrophenyl phosphate (9) in H(2) (18)O led to the formation of the (18)O-labeled O,O-diisopropyl phosphate product and confirmed that the catalytic reaction starts with cleavage of the P--O bond. From chemical and kinetic studies, the utilization of an optically pure S(p) enantiomer of O-methyl-p-nitrophenyl methylphosphonothioate (S(p))-MNMPT, 7) has demonstrated that the enzymatic reaction proceeds through a displacement mechanism and generates a chiral product in situ with an inversion of stereochemical configuration at the phosphorus atom. The results also lead to the conclusion that alteration of the active site through site-directed mutagenesis can result in a preference for (S(p))-MNMPT (7) rather than the R(p) isomer. PMID- 16470767 TI - Orientational control of guest molecules in an organic intercalation system by host polymer tacticity. AB - Four kinds of stereoregular poly(muconic acid)s, which are synthesized by topochemical polymerization and subsequent solid-state hydrolysis, are used as the organic host materials for intercalation. We describe the reaction behavior and layered structure of intercalation compounds using stereoregular poly(muconic acid)s and n-alkylamines as host and guest, respectively. The packing structure of the guest alkylamines was determined by X-ray diffractions as well as IR and Raman spectroscopies. We have found that the orientation of the guest molecules is controlled by the host polymer tacticity, depending on the structure of the two-dimensional hydrogen-bonding network formed in the polymer sheets of the crystals. PMID- 16470766 TI - The structural basis for docking in modular polyketide biosynthesis. AB - Polyketide natural products such as erythromycin and rapamycin are assembled on polyketide synthases (PKSs), which consist of modular sets of catalytic activities distributed across multiple protein subunits. Correct protein-protein interactions among the PKS subunits which are critical to the fidelity of biosynthesis are mediated in part by "docking domains" at the termini of the proteins. The NMR solution structure of a representative docking domain complex from the erythromycin PKS (DEBS) was recently solved, and on this basis it has been proposed that PKS docking is mediated by the formation of an intermolecular four-alpha-helix bundle. Herein, we report the genetic engineering of such a docking domain complex by replacement of specific helical segments and analysis of triketide synthesis by mutant PKSs in vivo. The results of these helix swaps are fully consistent with the model and highlight residues in the docking domains that may be targeted to alter the efficiency or specificity of subunit-subunit docking in hybrid PKSs. PMID- 16470768 TI - Interplay of p-sulfonatocalix[4]arene and crown ethers en route to molecular capsules and "Russian dolls". AB - Diffusion-ordered (1)H NMR spectroscopy techniques have been used to determine the binding strength of p-sulfonatocalix[4]arene (SO(3)[4]) towards a number of charged crown ether species in aqueous conditions. For several (doubly) charged (di)azacrown ethers, all were bound by SO(3)[4] either well or very well with binding constants between 5.1 x 10(2)-9.9 x 10(5) M(-1). These results correlate with, and thus explain the phenomenon of rapid capture of azacrown ethers in molecular capsules based on p-sulfonatocalix[4]arene and lanthanide metals. Similarly, the formation of "Russian doll" superanions in the solution phase is also elucidated. These superanions have been shown to selectively crystallise particular polynuclear aquated metal ions from mixtures in the aqueous phase. Neutral [18]crown-6 is not bound by p-sulfonatocalix[4]arene and displays a binding constant of 0 M(-1). When sodium [18]crown-6 is examined in a similar fashion, binding by SO(3)[4] is observed in solution with K(a) approximately 3.1 x 10(3) M(-1). PMID- 16470769 TI - Frontal polymerization synthesis of starch-grafted hydrogels: Effect of temperature and tube size on propagating front and properties of hydrogels. AB - The frontal polymerization process was used to produce superabsorbent hydrogels based on acrylic acid monomers grafted onto starch. Using a simple test tube which was nonadiabatic and permitted contact with air, the effects of initial temperature and tube size on the propagating front of grafting copolymerization and the properties of hydrogels were explored. The unrestricted access of the reaction mixture to oxygen delayed the formation of self-propagating polymerization front. The ignition time was markedly lengthened with the increasing of tube size attributed to the formation of large amounts of peroxy radicals. The front velocity dependence on initial temperature could be fit to an Arrhenius function with the average apparent activation energy of 24 kJ mol(-1), and on tube size to a function of higher order. The increase of the initial temperature increased the front temperature, which lead to more soluble oligomers and higher degree of crosslinking. The interplay of two opposite effects of oligomer and crosslinking determined the sol and gel content. An increase in tube size had two effects on the propagating front. One was to reduce heat loss. The other effect was to increase the number of escaping gas bubbles. The combined action of the two effects resulted in a maximum value of front temperature, an increase in sol content and a reduction in gel content with tube size. The highest swelling capacity of hydrogels was obtained when the initial temperature or tube size favored a formation of porous microstructure of hydrogels. PMID- 16470770 TI - Helical stacking tuned by alkoxy side chains in pi-conjugated triphenylbenzene discotic derivatives. AB - We report on the synthesis and self-assembly of a new series of discotic molecules containing triphenylbenzene as the core and alkoxy side chain with varying length. It was found that compounds 3 a-c, 4 b and 5 b could form stable gels in several apolar solvents. Transmission electron microscopy (TEM) images revealed that their morphologies were very different for the different alkoxy substituted organogels. In toluene or hexane, 3 b and 3 c resulted in both left- and right-handed helical fibers, whereas 3 a resulted in straight rigid fibers; 4 b and 5 b resulted in most straight fibers with a few twisted fibers. The results from FT-IR and UV/Vis absorption spectroscopy indicated that the hydrogen bonding and pi-pi interactions were the main driving forces for the formation of the self assembled gels. Further detailed analysis of their aggregation modes were conducted by UV-visible absorption spectra and X-ray diffraction (XRD) measurements. Based on these findings, the influence of these peripheral alkoxy substituents on the gel formation and the aggregation mode were discussed. The special enhanced fluorescent emissions, which resulted from aggregation, were also found in the gel phase. PMID- 16470771 TI - Glycopeptide synthesis through endo-glycosidase-catalyzed oligosaccharide transfer of sugar oxazolines: probing substrate structural requirement. AB - An array of sugar oxazolines was synthesized and tested as donor substrates for the Arthrobacter endo-beta-N-acetylglucosaminidase (Endo-A)-catalyzed glycopeptide synthesis. The experiments revealed that the minimum structure of the donor substrate required for Endo-A catalyzed transglycosylation is a Man beta1-->4-GlcNAc oxazoline moiety. Replacement of the beta-D-Man moiety with beta D-Glc, beta-D-Gal, and beta-D-GlcNAc monosaccharides resulted in the loss of substrate activity for the disaccharide oxazoline. Despite this, the enzyme could tolerate modifications such as attachment of additional sugar residues or a functional group at the 3- and/or 6-positions of the beta-D-Man moiety, thus allowing a successful transfer of selectively modified oligosaccharides to the peptide acceptor. On the other hand, the enzyme has a great flexibility for the acceptor portion and could take both small and large GlcNAc-peptides as the acceptor. The studies implicate a great potential of the endoglycosidase catalyzed transglycosylation for constructing both natural and selectively modified glycopeptides. PMID- 16470772 TI - Wire-type ruthenium(II) complexes with terpyridine-containing [2]rotaxanes as ligands: Synthesis, characterization, and photophysical properties. AB - [2]Rotaxanes based on the 1,2-bis(pyridinium)ethane subset[24]crown-8 ether motif were prepared that contain a terminal terpyridine group for coordination to a transition-metal ion. These rotaxane ligands were utilized in the preparation of a series of heteroleptic [Ru(terpy)(terpy-rotaxane)]2+ complexes. The compounds were characterized by 1D and 2D 1H NMR spectroscopy, X-ray crystallography, and high-resolution electrospray ionization mass spectrometry. The effect of using a rotaxane as a ligand was probed by UV/Vis/NIR absorption and emission spectroscopy of the Ru(II) complexes. In contrast with the parent [Ru(terpy)(2)]2+ complex, at room temperature the examined complexes exhibit a luminescence band in the near infrared region and a relatively long lived triplet metal-to-ligand charge-transfer (3MLCT) excited state, owing to the presence of strong-electron-acceptor pyridinium substituents on one of the two terpy ligands. Visible-light excitation of the Ru-based chromophore in acetonitrile at room temperature causes an electron transfer to the covalently linked 4,4' bipyridinium unit and the quenching of the MLCT luminescence. The 3MLCT excited state, however, is not quenched at all in rigid matrix at 77 K. The rotaxane structure was found to affect the absorption and luminescence properties of the complexes. In particular, when a crown ether surrounds the cationic axle, the photoinduced electron-transfer process is slowed down by a factor from 2 to 3. Such features, together with the synthetic and structural advantages offered by [Ru(terpy)2]2+-type complexes compared to, for example, [Ru(bpy)3]2+-type compounds, render these rotaxane-metal complexes promising candidates for the construction of photochemical molecular devices with a wire-type structure. PMID- 16470773 TI - Metal nuclearity modulated four-, six-, and eight-connected entangled frameworks based on mono-, bi-, and trimetallic cores as nodes. AB - To investigate the relationship between network connectivity and metal nuclearity, we designed and synthesized a series of three-dimensional (3D) entangled coordination frameworks based on different metal cores, namely [Zn(2)(bdc)(2)(L)(2)]2H(2)O (1), [Zn(bdc)(L)(0.5)] (2), [Zn(oba)(L)(0.5)] (3) and [Cd(3)(bdc)(3)(L)(2)(H(2)O)(2)] (4) by self-assembly of d(10) metal salts with the flexible long-chain ligand 1,4-bis(1,2,4-triazol-1-yl)butane (L), and with the rigid and nonrigid aromatic dicarboxylate ligands 1,4-benzenedicarboxylate (bdc) and 4,4'-oxybis(benzoate) (oba). Compound 1 exhibits a threefold interpenetrated diamondoid array typically based on a tetrahedral second building unit (SBU) at a single Zn center. Compound 2 adopts a threefold interpenetrated alpha-polonium-type network that is built from bimetallic cores as six-connected vertices. The structure of 3 also consists of dinuclear units; it comprises a novel (3,4)-connected threefold interpenetrated net with complex (4610)(46(2)10(3)) topology when single zinc centers act as four-connected nodes (or the alpha-Po topology if dinuclear units are considered as six-connected nodes). Compound 4, derived from a crosslinked fivefold interpenetrated diamond like substructure, is an unusual example of a self-penetrating coordination framework displaying an unprecedented eight-connected 4(20)6(8) topology with trinuclear cadmium clusters as eight-connected nodes which, to our knowledge, not only defines a new topology for eight-connected coordination networks, but also represents the highest connected topology presently known for self-penetrating systems. Detailed structural comparison of these complexes indicates that the increase in metal nuclearity induces the progressive increase in the connectivities of the ultimate nets: that is, the metal nuclearity plays a significant role in tuning the connectivity of a specific network. The thermal and luminescent properties of these compounds are discussed. PMID- 16470774 TI - Separation of cationic polymer particles and characterization of avidin immobilized particles by capillary electrophoresis. AB - Cationic polymer microparticles have received much attention especially in the field of biotechnology, such that their analysis and separation have become important. So far, the separation of cationic polymer particles with different size using CE has not been achieved and the cationic particles migrated as if they are negatively charged, probably due to electrostatic interaction between capillary wall and cationic polymer particles. In this paper, the separation of cationic polymer microparticles by CE was investigated in detail. The separation of cationic particles with different size was achieved in CE by taking into account the interaction between sample particles and the inner surface of capillaries. By employing a poly(vinyl alcohol)-coated capillary, a better size separation of amine-modified latex particles was obtained compared to a Polybrene coated capillary. It was elucidated that the composition, concentration, and pH of the background solution were also important factors in the separation of colloidal particles to avoid the surface adsorption and the characteristic aggregation of polymer particles. Furthermore, the CE analysis was applied to the characterization of cationic protein-immobilized particles. PMID- 16470775 TI - Electrophoresis of DNA in human genetic diagnostics - state-of-the-art, alternatives and future prospects. AB - Electrophoretic separation of nucleic acids according to their molecular weights has dominated the methods' spectrum in molecular genetics for nearly half a century. We review the current methodological basis and evaluate its impact with special reference to new developments in the microarray technology. Although electrophoresis may be made redundant for many applications in DNA diagnostics within a few years, a number of electrophoretic vestiges will remain irreplaceable in the foreseeable future. PMID- 16470776 TI - Proteome analysis of human plasma and amniotic fluid by Off-Gel isoelectric focusing followed by nano-LC-MS/MS. AB - This paper presents a comparative proteomic analysis of human maternal plasma and amniotic fluid (AF) samples from the same patient at term of pregnancy in order to find specific AF proteins as markers of premature rupture of membranes, a complication frequently observed during pregnancy. Maternal plasma and the corresponding AF were immunodepleted in order to remove the six most abundant proteins before the systematic analysis of their protein composition. The protein samples were then fractionated by IEF Off-Gel electrophoresis (OGE), digested and analyzed with nano-LC-MS/MS separation, revealing a total of 73 and 69 proteins identified in maternal plasma and AF samples, respectively. The proteins identified in AF have been compared to those identified in the mother plasma as well as to the reference human plasma protein list reported by Anderson et al. (Mol. Cell. Proteomics 2004, 3, 311-326). This comparison showed that 26 proteins were exclusively present in AF and not in plasma among which 10 have already been described to be placenta or pregnancy specific. As a further validation of the method, plasma proteins fractionated by OGE and analysed by nano-LC-MS/MS have been compared to the Swiss 2-D PAGE reference map by reconstructing a map that matches 2-D gel and OGE experimental data. This representation shows that 36 of 49 reference proteins could be identified in both data sets, and that isoform shifts in pI are well conserved in the OGE data sets. PMID- 16470777 TI - Measurement of specific radioactivity in proteins separated by two-dimensional gel electrophoresis. AB - We report a method to quantify the specific radioactivity of proteins that have been separated by 2-DE. Gels are stained with SyproRuby, and protein spots are excised. The SyproRuby dye is extracted from each spot using DMSO, and the fluorescence is quantified automatically using a plate reader. The extracted gel piece is then dissolved in hydrogen peroxide and radioactivity is quantified by liquid scintillation counting. Gentle agitation with DMSO for 24 h was found to extract all the SyproRuby dye from gel fragments. The fluorescence of the extract was linearly related to the amount of BSA loaded onto a series of 1-D gels. When rat muscle samples were run on 2-DE gels, the fluorescence extracted from 54 protein spots showed a good correlation (r = 0.79, p < 0.001) with the corresponding spot intensity measured by conventional scanning and image analysis. DMSO extraction was found not to affect the amount of radioactive protein left in the gel. When a series of BSA solutions of known specific radioactivity were run on 2-DE gels, the specific radioactivity measured by the new method showed a good correlation (r = 0.98, p < 0.01, n = 5) with the specific radioactivity measured directly before loading. Reproducibility of the method was measured in a series of 2-DE gels containing proteins from the livers of rats and mice that had been injected with [35S]methionine. Variability tended to increase when the amount of radioactivity in the protein spot was low, but for samples containing at least 10 dpm above background the CV was around 30%, which is comparable to that obtained when measuring protein expression by conventional image analysis of SyproRuby-stained 2-DE gels. Similar results were obtained whether spots were excised manually or using a spot excision robot. This method offers a high-throughput, cost-effective and reliable method of quantifying the specific radioactivity of proteins from metabolic labelling experiments carried out in vivo, so long as sufficient quantities of radioactive tracer are used. PMID- 16470779 TI - Parallel analysis of biomolecules on a microfabricated capillary array chip. AB - This paper focused on a self-developed microfluidic array system with microfabricated capillary array electrophoresis (mu-CAE) chip for parallel chip electrophoresis of biomolecules. The microfluidic array layout consists of two common reservoirs coupled to four separation channels connected to sample injection channel on the soda-lime glass substrate. The excitation scheme for distributing a 20 mW laser beam to separation channels in an array is achieved. Under the control of program, the sample injection and separation in multichannel can be achieved through six high-voltage modules' output. A CCD camera was used to monitor electrophoretic separations simultaneously in four channels with LIF detection, and the electropherograms can be plotted directly without reconstruction by additional software. Parallel multichannel electrophoresis of series biomolecules including amino acids, proteins, and nucleic acids was performed on this system and the results showed fine reproducibility. PMID- 16470778 TI - Coordinated and reversible reduction of enzymes involved in terminal oxidative metabolism in skeletal muscle mitochondria from a riboflavin-responsive, multiple acyl-CoA dehydrogenase deficiency patient. AB - In this case report we studied alterations in mitochondrial proteins in a patient suffering from recurrent profound muscle weakness, associated with ethylmalonic adipic aciduria, who had benefited from high dose of riboflavin treatment. Morphological and biochemical alterations included muscle lipid accumulation, low muscle carnitine content, reduction in fatty acid beta-oxidation and reduced activity of complexes I and II of the respiratory chain. Riboflavin therapy partially or totally reversed these symptoms and increased the level of muscle flavin adenine dinucleotide, suggesting that aberrant flavin cofactor metabolism accounted for the disease. Proteomic investigation of muscle mitochondria revealed decrease or absence of several flavoenzymes, enzymes related to flavin cofactor-dependent mitochondrial pathways and mitochondrial or mitochondria associated calcium-binding proteins. All these deficiencies were completely rescued after riboflavin treatment. This study indicates for the first time a profound involvement of riboflavin/flavin cofactors in modulating the level of a number of functionally coordinated polypeptides involved in fatty acyl-CoA and amino acid metabolism, extending the number of enzymatic pathways altered in riboflavin-responsive multiple acyl-CoA dehydrogenase deficiency. PMID- 16470780 TI - A new type of migrating zone boundary in electrophoresis: 3. The hybrid boundary and stacking criteria. AB - This article investigates the principles of stacking of analytes by a zone of a bulk sample component that possesses one hybrid boundary. The model system investigated comprises a transient stacking zone in a BGE where the rear boundary of this zone is a hybrid one. A theoretical description of such a system is given, and general rules and mathematical criteria for quantitative stacking are presented. These criteria are based on comparison of migration velocities of analytes to be stacked and velocities of the boundaries of the stacking zone. It is shown that the presence of a hybrid boundary brings about additional constraints for stacking of analytes due to the presence of the sharp part of the hybrid boundary that migrates faster than regular sharp boundary of this zone. An experimental example related to the described theory is also shown. PMID- 16470781 TI - Capillary electrophoresis using copolymers of different composition as physical coatings: a comparative study. AB - In this work, a comparative study on the use of different polymers as physically adsorbed coatings for CE is presented. It is demonstrated that the use of ad hoc synthesized polymers as coatings allows tailoring the EOF in CE increasing the flexibility of this analytical technique. Namely, different polymers were synthesized at our laboratory using different percentages of ethylpyrrolidine methacrylate (EpyM) and N,N-dimethylacrylamide (DMA). Thus, by modifying the percentage of EpyM and DMA monomers it is possible to manipulate the positive charge of the copolymer, varying the global electrical charge on the capillary wall and with that the EOF. These coated capillaries are obtained by simply flushing a given EpyM-DMA aqueous solution into bare silica capillaries. It is shown that by using these coated capillaries at adequate pHs, faster or more resolved CE separations can be achieved depending on the requirements of each analysis. Moreover, it is demonstrated that these coated capillaries reduce the electrostatic adsorption of basic proteins onto the capillary wall. Furthermore, EpyM-DMA coatings allow the reproducible chiral separation of enantiomers through the partial filling technique (PFT). The EpyM-DMA coated capillaries are demonstrated to provide reproducible EOF values independently of the pH and polymer composition with%RSD values lower than 2% for the same day. It is also demonstrated that the coating procedure is reproducible between capillaries. The compatibility of this coating protocol with CE in microchips is discussed. PMID- 16470782 TI - Theoretical and empirical approaches to express the mobility of small ions in capillary electrophoresis. AB - A discussion is given about the concepts of the ion mobility, the analyte property which governs migration and thus separation selectivity in CE. It deals with small organic and inorganic ions, not with charged polymers or large particles like colloids. The discussion is directed to two main concepts. (i) The first is based on physico-chemistry of ion conductance in solution, and distinguishes three types of mobility. The absolute mobility is the limiting mobility at zero ionic strength; it depends on the solvent and the temperature. It is obtained by extrapolation of the actual mobilities, those of the fully charged particles at finite ion concentration. The observed reduction of the absolute mobility with ionic concentration is related to an ion cloud, and is formulated by the established theories of ion conductance. It explains the actual mobility as function of (beside other factors) the ionic strength, the viscosity and relative permittivity of the solvent, the temperature, the relaxation time of solvent polarisation and the distance of closest approach between ion and counterion. The effective mobility, finally, is the mobility when association and dissociation equilibria play a role. Most important are acid-base reactions, but complexation, ion pairing and homo- and heteroconjugation were considered as well. (ii) The second approach treats mobility data with different mathematical methods, and formulates their dependence on variables like solvent composition with appropriate algorithms. These empirical methods mainly include least squares and neural network-based methods. The least square methods ranges from the simplest model, which uses only the molecular weight of the analyte, to more complicated model requiring three-dimensional structural descriptors of the solutes. Neural networks have been applied to model the mobility using different input variables and various architectures. Work comparing the accuracy of least squares and neural network methods was discussed; the results showed that the neural network method leads to a more accurate mobility calculation. However, the least squares methods could give some information to the factors affecting the mobility of the analytes. The resulting methods allow the prediction of mobilities under different experimental conditions with certain accuracy. It has been shown that using such models, it is possible to predict mobility of analytes after training the models by a minimum number of data to speed up the method development stage. PMID- 16470783 TI - A novel specific heparin-binding activity of bovine folate-binding protein characterized by capillary electrophoresis. AB - Folate-binding proteins (FBPs) are ubiquitous, soluble and membrane-bound high affinity receptors for folate, an essential nutrient involved in nucleic and amino acid metabolism. In the course of optimizing CE separation conditions for FBP purified from cow's milk we discovered a novel specific heparin-binding activity of FBP by affinity CE. Heparin is a highly sulfated glycosaminoglycan and thus prone to induce anodic migration shifts of complexing analytes. Prior complexation of FBP with folate abolished heparin binding, and thus folate competes with heparin for binding to FBP. It was estimated that heparin bound several orders of magnitude less strongly than folate with an average dissociation constant in the 1-10 microM range. In contrast to the mobility shifts induced by heparin, free and folate-bound FBP were not separated by CE. However, binding of folate induced a distinct increase in FBP-peak symmetry, and using heparin as an affinity displacer, the free FBP in equilibrium with folate FBP complexes could readily be separated from the complexes. While the folate-FBP interaction was too strong to be characterized quantitatively because of inadequate detection limits of a UV-based detection system, it was possible to estimate the folate-FBP binding stoichiometry using this approach. The heparin interaction fractionated FBP into distinct subfractions, and the CE approach thus promises to be useful for unraveling the complex oligomerization behavior of FBP isoforms as well as for evaluating the FBP affinity for various species and analogs of glycosaminoglycans and folate. PMID- 16470784 TI - Affinity monolithic capillary columns for glycomics/proteomics: 1. Polymethacrylate monoliths with immobilized lectins for glycoprotein separation by affinity capillary electrochromatography and affinity nano-liquid chromatography in either a single column or columns coupled in series. AB - In this report, microcolumn separation schemes involving monolithic capillary columns with immobilized lectins, and relevant to nanoglycomics/nanoproteomics were introduced. Positive and neutral monoliths based on poly(glycidyl methacrylate-co-ethylene dimethacrylate) were designed for achieving lectin affinity chromatography (LAC) by nano-LC and CEC. The positive monoliths (i.e., monoliths with cationic sites) afforded relatively high permeability in nano-LC but lack predictable EOF magnitude and direction, while neutral monoliths provided a good compromise between reasonable permeability in nano-LC and predictable EOF in CEC. Lectin affinity nano-LC permitted the enrichment of classes of different glycoproteins having similar N-glycans recognized by the immobilized lectin, whereas lectin affinity CEC provided the simultaneous capturing and separation of different glycoproteins due to differences in charge to-mass ratio. Also, this investigation demonstrated for the first time the coupling of lectin capillary columns in series (i.e., tandem columns) for enhanced separation of glycoproteins by LAC using the CEC modality. Furthermore, in the coupled columns format, glycoforms of a given glycoprotein were readily separated. PMID- 16470785 TI - Proteases and chaperones are the most abundant proteins in the parasitophorous vacuole of Plasmodium falciparum-infected erythrocytes. AB - After invasion of erythrocytes, the human malaria parasite Plasmodium falciparum resides within a parasitophorous vacuole (PV) which forms an interface between the host cell cytosol and the parasite surface. This vacuole protects the parasite from potentially harmful substances, but allows access of essential nutrients to the parasite. Furthermore, the vacuole acts as a transit compartment for parasite proteins en route to the host cell cytoplasm. Recently we developed a strategy to biotin label soluble proteins of the PV. Here, we have paired this strategy with a high-throughput MALDI-TOF-MS analysis to identify 27 vacuolar proteins. These proteins fall into the following main classes: chaperones, proteases, and metabolic enzymes, consistent with the expected functions of the vacuole. These proteins are likely to be involved in several processes including nutrient acquisition from the host cytosol, protein sorting within the vacuole, and release of parasites at the end of the intraerythrocytic cycle. PMID- 16470786 TI - Crossroads in GDNF therapy for Parkinson's disease. AB - The development of a neuroprotective or neuroregenerative therapy for Parkinson's disease (PD) would be a major therapeutic advance. Unfortunately, results from a recent controlled clinical study delivering the neurotrophic factor, glial derived neurotrophic factor (GDNF), directly into brain did not demonstrate efficacy and safety of such a treatment. A critical review of available data suggests that there are questions that need to be answered before the future of GDNF as a therapy for PD can be determined. PMID- 16470788 TI - The biallelic expression pattern of X-linked genes in Klinefelter syndrome by pyrosequencing. PMID- 16470787 TI - Genotype-phenotype correlation in hereditary hemorrhagic telangiectasia: mutations and manifestations. AB - Hereditary hemorrhagic telangiectasia (HHT) is a genetically heterogeneous vascular dysplasia with multiple telangiectases and arteriovenous malformations and it is caused by mutations in endoglin gene (ENG) (HHT1) and activin A receptor type II-like 1 gene (ACVRL1) (HHT2). We evaluated 111 patients with HHT from 34 families by history, examination, screening for vascular malformations, and sequencing of both genes. We found mutations in 26 of the 34 kindreds (76%) analyzed-54% were in ENG and 46% were in ACVRL1. Mutations in ACVRL1 cluster largely in exons 7 and 8, but ENG mutations were widely distributed within that gene. We found that epistaxis had an earlier onset in patients with HHT1 than those with HHT2, but the severity by middle ages was similar. Pulmonary arteriovenous malformations were more frequent and on the average of larger size in HHT1. Hepatic vascular malformations were more common in patients with HHT2. Cerebral arteriovenous malformations were more common in patients with HHT1, but spinal arteriovenous malformations were seen only in patients with HHT2. Truncating mutations in ENG were associated with more affected organs and more severe hemorrhaging than were missense mutations. We conclude that HHT2 has a later onset than HHT1 and the former may disproportionately involve smaller vessels in tissues with more significant vascular remodeling. PMID- 16470789 TI - Characterization of two supernumerary marker chromosomes in a patient with signs of Klinefelter syndrome, mild facial anomalies, and severe speech delay. AB - A boy with signs of Klinefelter syndrome, mild facial dysmorphic features, and severely retarded speech development displayed a female karyotype with mosaicism for two marker chromosomes 48,XX,+mar1,+mar2[68]/47,XX,+mar1[19]/47,XX,+mar2[6]/46,XX[8]. Using chromosomal microdissection, locus-specific fluorescence in situ hybridization (FISH), and PCR with several Y-chromosome markers, the larger supernumerary marker chromosome (SMC) was characterized as a ring Y-chromosome. Detection of the SRY-region explained the male phenotype. The smaller second marker chromosome contained the pericentromeric region of chromosome 8. We suggest that the co-occurrence of a partial Y-chromosome and partial trisomy 8 explain the severe speech delay and the facial dysmorphic features. PMID- 16470790 TI - Molecular cytogenetic analysis of a de novo interstitial deletion of 5q23.3q31.2 and its phenotypic consequences. AB - We report a 2(3/12)-year-old boy with a constitutional interstitial deletion of 5q,46,XY,del(5)(q23.3q31.2) de novo. Clinical manifestations in this patient included failure to thrive, psychomotor retardation, mild facial dysmorphic features, and long and slender fingers and toes. The precise location and extent (9.5 Mb) of the deletion was determined by fluorescence in situ hybridization (FISH) using 19 YAC and BAC clones. Comparison of the present patient with six other patients with deletions of chromosomal bands 5q22-5q31 allowed further delineation of a constitutional del5q22q31 syndrome. The main features of this syndrome are psychomotor retardation, failure to thrive, hypotonia, hypoplastic muscles, cleft or high arched palate, low-set and dysplastic ears, flat nasal bridge, downslanting palpebral fissures, hypertelorism, anteverted nostrils, and micro- and/or retrognathia. PMID- 16470791 TI - Case of chromosome 6p25 terminal deletion associated with Axenfeld-Rieger syndrome and persistent hyperplastic primary vitreous. AB - Axenfeld-Rieger syndrome is inherited in an autosomal dominant pattern and is characterized by anomalies of the anterior segment of the eye and systemic signs including craniofacial dysmorphic features and cardiac defects. The disorder is genetically heterogeneous and one causative gene, FOXC1, is located on chromosome 6p25. Persistent hyperplastic primary vitreous (PHPV) is a congenital ocular disorder in which there is a failure of the normal regression of the primary vitreous and a proliferation of fibrous tissue from the remnants of the primary vitreous. Deletions of chromosome 6p25 have been reported in a small number of patients with Axenfeld-Rieger syndrome; however, no case of chromosome 6p25 deletion has been reported with PHPV. We report a newborn girl who had both Axenfeld-Rieger syndrome and the combined type of PHPV, in whom the G-banding and spectral karyotyping revealed a 6p monosomy of terminal deletion with a breakpoint at chromosome 6p25.1. The karyotype was 46,XX,del(6)(p25.1). We conclude that PHPV in the context of Axenfeld-Rieger syndrome can be caused by 6p25 terminal deletion. PMID- 16470792 TI - Klinefelter syndrome and mediastinal germ cell tumors. AB - Precocious puberty is not a typical manifestation of patients with Klinefelter syndrome (KS). However, there is an increased incidence of mediastinal germ cell tumors (M-GCT) in KS, whereas the discussion of a generally higher tumor risk in this condition is still controversial. A rare subgroup of KS patients consists of prepubertal children with precocious puberty due to human chorionic gonadotropin (hCG)-producing M-GCTs. We present clinical data on a boy with KS and sexual precocity, and summarize the published data on 12 boys with KS out of 54 cases of KS and M-GCT. CLINICAL REPORT: an 8.5-year-old boy presented with signs of precocious puberty. Laboratory analyses (suppressed gonadotropins, elevated testosterone) and thoracic CT demonstrated a beta-human chorionic gonadotropin (beta-hCG) and alpha(1)-feto protein (alpha-FP) secreting mediastinal tumor. Histological analysis showed a mixed germ cell tumor comprising choriocarcinoma (CH), embryonal carcinoma (EC), mature teratoma (MT), and yolk sac tumor (YS). He was successfully treated by surgery and adjuvant chemotherapy. Epianalysis of published cases: all KS patients (n = 12), age 4-9 years, presented with precocious sexual development (PP), whereas the older ones showed thorax associated symptoms, mainly chest pain, dyspnea, and cough. The histological distribution was also age-dependent with mixed germ cell tumors predominantly in younger patients. Thus, M-GCTs are strongly associated with precocious puberty in young boys with KS. Therefore, a karyotype analysis should be included in the clinical work-up of boys with precocious puberty and M-GCT. There is still no convincing explanation for the association of M-GCTs and KS. PMID- 16470793 TI - A truncating mutation in the IL1RAPL1 gene is responsible for X-linked mental retardation in the MRX21 family. AB - X-linked mental retardation (XLMR) is a genetically heterogeneous condition, due to mutations in at least 50 genes, involved in functioning of the central nervous system and located on the X chromosome. Nonspecific XLMR (MRX) is characterized essentially by mental retardation transmitted by X-linked inheritance. More than 80 extended MRX pedigrees have been reported to date, which have been distinguished exclusively by physical position of the corresponding gene on the X chromosome, established by linkage analysis. One such family, MRX21, which was described by us in 1993 and localized to Xp11.4-pter, has now been reanalyzed with additional markers and after one more affected individual had became available. This extra information allowed a significant reduction of the linkage interval and, eventually, identification of the mutant gene. A stop mutation in exon 10 of the IL1RAPL1 gene (in Xp21) was found in the four affected males and in obligate carriers, allowing conclusive counseling of other family members of uncertain carrier status. The W487X mutation results in the production of a truncated IL1RAPL protein, comprised of the extracellular Ig-like domain and transmembrane tract, but lacking the last 210 aminoacids of the cytoplasmic domain. MRX21 is the first extended MRX family with a point mutation in IL1RAPL1 and the second with a stop mutation, which had been previously found only in a small family. Our report confirms the role of the IL1RAPL1 gene in causing nonspecific mental retardation in males and underlines the importance of detailed linkage analysis before candidate gene mutational screening. PMID- 16470794 TI - Speech and language impairment and oromotor dyspraxia due to deletion of 7q31 that involves FOXP2. AB - We report detailed clinical, cytogenetic, and molecular findings in a girl with a deletion of chromosome 7q31-q32. This child has a severe communication disorder with evidence of oromotor dyspraxia, dysmorphic features, and mild developmental delay. She is unable to cough, sneeze, or laugh spontaneously. Her deletion is on the paternally inherited chromosome and includes the FOXP2 gene, which has recently been associated with speech and language impairment and a similar form of oromotor dyspraxia in at least three other published cases. We hypothesize that our patient's communication disorder and oromotor deficiency are due to haploinsufficiency for FOXP2 and that her dysmorphism and developmental delay are a consequence of the absence of the other genes involved in the microdeletion. We propose that this patient, together with others reported in the literature, may define a new contiguous gene deletion syndrome encompassing the 7q31-FOXP2 region. Cytogenetic and molecular analysis of this region should be considered for other individuals displaying similar characteristics. PMID- 16470796 TI - Chimerism in twins: caution is needed in interpretation of karyotypes. PMID- 16470795 TI - Extended mutational analyses of FGFR1 in osteoglophonic dysplasia. PMID- 16470797 TI - POR R457H is a global founder mutation causing Antley-Bixler syndrome with autosomal recessive trait. PMID- 16470798 TI - Recurrence of SOX2 anophthalmia syndrome with gonosomal mosaicism in a phenotypically normal mother. PMID- 16470799 TI - Metal enolates of alpha-CF3 ketones: theoretical guideline, direct generation, and synthetic use. AB - The difficulty as well as the significance of the direct generation of metal enolates of alpha-CF(3) ketones cannot be easily understood by chemists unfamiliar with F. In sharp contrast to the sunny side of non-F, hydrocarbon chemistry, F chemistry has long been overshadowed. Metal enolates of carbonyl compounds are synthetically important in C-C bond-forming reactions. However, the metal enolates of fluorinated carbonyl compounds have been severely limited to alpha-F metal enolates. Alpha-CF(3) metal enolates have generally been recognized as unstable and difficult to prepare because of the facile beta-M-F elimination. However, we have developed a direct generation and synthetic application of alpha CF(3) metal enolates. Therefore, the present results regarding the direct generation and synthetic use of metal enolates of alpha-CF(3) ketones might be recognized as a real breakthrough for the general use of metal enolates in F chemistry. PMID- 16470800 TI - Polyoxometalate catalysts: toward the development of green H2O2-based epoxidation systems. AB - This paper describes the development of green, efficient H(2)O(2)-based epoxidation systems with three kinds of polyoxometalates: (i) a dinuclear peroxotungstate [W(2)O(3)(O(2))(4)(H(2)O)(2)](2-) (I), (ii) a divacant lacunary polyoxotungstate [gamma-SiW(10)O(34)(H(2)O)(2)]4 (II), (iii) and a divanadium substituted polyoxotungstate [gamma-1,2-H(2)SiV(2)W(10)O(40)](4-) (III). The highly chemo-, regio-, and diastereoselective epoxidation of various allylic alcohols with only 1 equiv H(2)O(2) in water can be efficiently catalyzed by potassium salt of I (K-I). The catalyst K-I can be recycled with the retention of the catalytic performance. Protonation of a divacant lacunary polyoxotungstate [gamma-SiW(10)O(36)](8-) gives [gamma-SiW(10)O(34)(H(2)O)(2)](4-) (II) with two aquo ligands. The tetra-n-butylammonium salt of II (TBA-II) catalyzes epoxidation of common olefins including propylene with >or=99% selectivity to epoxide and >or=99% efficiency of H(2)O(2) utilization. The bis(mu-hydroxo)bridged dioxovanadium site in [gamma-1,2-H(2)SiV(2)W(10)O(40)](4-) (III) can also efficiently catalyze epoxidation of a variety of olefins with 1 equiv H(2)O(2). Notably, the system with III shows unique stereospecificity, diastereoselectivity, and regioselectivity for the epoxidation of cis/trans olefins, 3-substituted cyclohexenes, and nonconjugated dienes, respectively, which are quite different from those reported for epoxidation systems up to now. Furthermore, the heterogenization of the mentioned polyoxometalates can be achieved by using ionic liquid-modified SiO(2) as a support without loss of catalytic performance. PMID- 16470801 TI - Controlling the regiochemistry of radical cyclizations. AB - This review describes the results of our recent studies on the control of the regiochemistry of radical cyclizations. N-vinylic alpha-chloroacetamides generally cyclized in a 5-endo-trig manner to give five-membered lactams, whereas 4-exo-trig cyclization occurred when the cyclized radical intermediates were highly stabilized by an adjacent phenyl or phenylthio group to afford beta lactams. The 5-exo or 6-exo cyclization of aryl radicals onto the alkenic bond of enamides could be shifted to the corresponding 6-endo or 7-endo mode of cyclization by a positional change of the carbonyl group of enamides. The 6-endo- and 7-endo-selective aryl radical cyclizations were applied to radical cascades for the synthesis of alkaloids such as phenanthroindolizidine, cephalotaxine skeleton, and lennoxamine. The 5-exo-trig cyclization of an alkyl radical onto the alkenyl bond of enamides could also be shifted to the 6-endo mode by a positional change of the carbonyl group of enamides. The 6-endo- selective cyclization was applied to the radical cascade to afford a cylindricine skeleton. Other examples of controlling the regiochemistry of radical cyclizations and their applications to the synthesis of natural products are also discussed. PMID- 16470802 TI - Organic-inorganic hybrid mesoporous silicas: functionalization, pore size, and morphology control. AB - Topological design of mesoporous silica materials, pore architecture, pore size, and morphology are currently major issues in areas such as catalytic conversion of bulky molecules, adsorption, host-guest chemistry, etc. In this sense, we discuss the pore size-controlled mesostructure, framework functionalization, and morphology control of organic-inorganic hybrid mesoporous silicas by which we can improve the applicability of mesoporous materials. First, we explain that the sizes of hexagonal- and cubic-type pores in organic-inorganic hybrid mesoporous silicas are well controlled from 24.3 to 98.0 A by the direct micelle-control method using an organosilica precursor and surfactants with different alkyl chain lengths or triblock copolymers as templates and swelling agents incorporated in the formed micelles. Second, we describe that organic-inorganic hybrid mesoporous materials with various functional groups form various external morphologies such as rod, cauliflower, film, rope, spheroid, monolith, and fiber shapes. Third, we discuss that transition metals (Ti and Ru) and rare-earth ions (Eu(3+) and Tb(3+)) are used to modify organic-inorganic hybrid mesoporous silica materials. Such hybrid mesoporous silica materials are expected to be applied as excellent catalysts for organic reactions, photocatalysis, optical devices, etc. PMID- 16470803 TI - Photorefractive effect of ferroelectric liquid crystals. AB - This paper reviews our recent work on the photorefractive effect of ferroelectric liquid crystals (FLCs). The photorefractive effect is defined as the optical modulation of the refractive index of a medium as a result of a variety of processes. The interference of two laser beams in a photorefractive material establishes a refractive index grating. This phenomenon enables the creation of different types of photonic applications. FLCs exhibit fast electric field response, and the orientation of the molecular axis of FLCs changes its direction according to the change in direction of the spontaneous polarization (Ps). When two laser beams interfere in a photoconductive FLC, an orientational grating is formed. The mechanism of the formation of the grating is based on the response of the Ps to the photoinduced internal electric field. The time of formation of the refractive index grating is significantly shorter in FLC materials. PMID- 16470804 TI - Regional cerebral brain metabolism correlates of neuroticism and extraversion. AB - Factor-analytic approaches to human personality have consistently identified several core personality traits, such as Extraversion/Introversion, Neuroticism, Agreeableness, Consciousness, and Openness. There is an increasing recognition that certain personality traits may render individuals vulnerable to psychiatric disorders, including anxiety disorders and depression. Our purpose in this study was to explore correlates between the personality dimensions neuroticism and extraversion as assessed by the NEO Five-Factor Inventory (NEO-FFI) and resting regional cerebral glucose metabolism (rCMRglu) in healthy control subjects. Based on the anxiety and depression literatures, we predicted correlations with a network of brain structures, including ventral and medial prefrontal cortex (encompassing anterior cingulate cortex and orbitofrontal cortex), insular cortex, anterior temporal pole, ventral striatum, and the amygdala. Twenty healthy women completed an (18F)FDG (18F-fluorodeoxyglucose) positron emission tomography (PET) scan at rest and the NEO-FFI inventory. We investigated correlations between scores on NEO-FFI Neuroticism and Extraversion and rCMRglu using statistical parametric mapping (SPM99). Within a priori search territories, we found significant negative correlations between Neuroticism and rCMRglu in the insular cortex and positive correlations between Extraversion and rCMRglu in the orbitofrontal cortex. No significant correlations were found involving anterior cingulate, amygdala, or ventral striatum. Neuroticism and Extraversion are associated with activity in insular cortex and orbitofrontal cortex, respectively. PMID- 16470805 TI - The fragment transformation method to detect the protein structural motifs. AB - To identify functional structural motifs from protein structures of unknown function becomes increasingly important in recent years due to the progress of the structural genomics initiatives. Although certain structural patterns such as the Asp-His-Ser catalytic triad are easy to detect because of their conserved residues and stringently constrained geometry, it is usually more challenging to detect a general structural motifs like, for example, the betabetaalpha-metal binding motif, which has a much more variable conformation and sequence. At present, the identification of these motifs usually relies on manual procedures based on different structure and sequence analysis tools. In this study, we develop a structural alignment algorithm combining both structural and sequence information to identify the local structure motifs. We applied our method to the following examples: the betabetaalpha-metal binding motif and the treble clef motif. The betabetaalpha-metal binding motif plays an important role in nonspecific DNA interactions and cleavage in host defense and apoptosis. The treble clef motif is a zinc-binding motif adaptable to diverse functions such as the binding of nucleic acid and hydrolysis of phosphodiester bonds. Our results are encouraging, indicating that we can effectively identify these structural motifs in an automatic fashion. Our method may provide a useful means for automatic functional annotation through detecting structural motifs associated with particular functions. PMID- 16470806 TI - Involvement of disulfide bonds and histidine 172 in a unique beta-sheet to alpha helix transition of alpha 1-acid glycoprotein at the biomembrane interface. AB - Human alpha(1)-acid glycoprotein (AGP), which is comprised of 183 amino acid residues and 5 carbohydrate chains, is a major plasma protein that binds to basic and neutral drugs as well as to steroid hormones. It has a beta-sheet-rich structure in aqueous solution. Our previous findings suggest that AGP forms an alpha-helix structure through an interaction with biomembranes. We report herein on a study of the mechanism of alpha-helix formation in AGP using various modified AGPs. The disulfide reduced AGP (R-AGP) was extensively unfolded, whereas asialylated AGP (A-AGP) maintained the native structure. Intriguingly, reduced and asialylated AGP (RA-AGP) increased the alpha-helix content as observed in the presence of biomembrane models, and showed a significant decrease in ligand binding capacity. This suggests that AGP has an innate tendency to form an alpha-helix structure, and disulfide bonds are a key factor in the conformational transition between the beta-sheet and alpha-helix structures. However, RA-AGP with all histidine residues chemically modified (HRA-AGP) was found to lose the intrinsic ability to form an alpha-helix structure. Furthermore, disulfide reduction of the H172A mutant expressed in Pichia pastoris also caused a similar loss of folding ability. The present results indicate that disulfide bonds and the C-terminal region, including H172 of AGP, play important roles in alpha-helix formation in the interaction of the protein with biomembranes. PMID- 16470807 TI - Fish consumption and breast cancer risk. The European Prospective Investigation into Cancer and Nutrition (EPIC). AB - There is current interest in fish consumption and marine omega-3 (n-3) fatty acids and breast cancer risk. Some in vitro and animal studies have suggested an inhibitory effect of marine n-3 fatty acids on breast cancer growth, but the results from epidemiological studies that have examined the association between fish consumption and breast cancer risk in humans are inconsistent. We examined fish consumption and breast cancer risk in 310,671 women aged between 25 and 70 yr at recruitment into the European Prospective Investigation Into Cancer and Nutrition (EPIC). The participants completed a dietary questionnaire between 1992 98 and were followed up for incidence of breast cancer for a median of 6.4 yr. Hazard ratio for breast cancer by intake of total and lean and fatty fish were estimated, stratified by study centre and adjusted for established breast cancer risk factors. During follow-up, 4,776 invasive incident breast cancers were reported. No significant associations between intake of total fish and breast cancer risk were observed, hazard ratio (HR) 1.01 (95% confidence interval [CI] 0.99-1.02; p = 0.28 per 10 g fish/day). When examining lean and fatty fish separately, we found a positive significant association only in the highest quintile for fatty fish (HR 1.13, 95% CI 1.01-1.26), but test for trend was not significant (p = 0.10). No associations with breast cancer risk were observed when the study participants were subdivided by menopausal status. Although the period of follow-up is relatively short, the results provide no evidence for an association between fish intake and breast cancer risk. PMID- 16470808 TI - Increased seizure duration and slowed potassium kinetics in mice lacking aquaporin-4 water channels. AB - The glial water channel aquaporin-4 (AQP4) has been hypothesized to modulate water and potassium fluxes associated with neuronal activity. In this study, we examined the seizure phenotype of AQP4 -/- mice using in vivo electrical stimulation and electroencephalographic (EEG) recording. AQP4 -/- mice were found to have dramatically prolonged stimulation-evoked seizures after hippocampal stimulation compared to wild-type controls (33 +/- 2 s vs. 13 +/- 2 s). In addition, AQP4 -/- mice were found to have a higher seizure threshold (167 +/- 17 microA vs. 114 +/- 10 microA). To assess a potential effect of AQP4 on potassium kinetics, we used in vivo recording with potassium-sensitive microelectrodes after direct cortical stimulation. Although there was no significant difference in baseline or peak [K(+)](o), the rise time to peak [K(+)](o) (t(1/2), 2.3 +/- 0.5 s) as well as the recovery to baseline [K(+)](o) (t(1/2), 15.6 +/- 1.5 s) were slowed in AQP4 -/- mice compared to WT mice (t(1/2), 0.5 +/- 0.1 and 6.6 +/- 0.7 s, respectively). These results implicate AQP4 in the expression and termination of seizure activity and support the hypothesis that AQP4 is coupled to potassium homeostasis in vivo. PMID- 16470809 TI - Enhancement of myoblast microencapsulation for gene therapy. AB - One method of nonviral-based gene therapy is to implant microencapsulated nonautologous cells genetically engineered to secrete the desired gene products. Encapsulating the cells within a biocompatible permselective hydrogel, such as alginate-poly-L-lysine-alginate (APA), protects the foreign cells from the host immune system while allowing diffusion of nutrients and the therapeutic gene products. An important consideration is which kind of cells is the best candidate for long-term implantation. Our previous work has shown that proliferation and differentiation of encapsulated C2C12 myoblasts in vitro are significantly improved by inclusion of basic fibroblast growth factor (bFGF), insulin growth factor II (IGF-II), and collagen within the microcapsules ("enhanced" capsules). However, the effects of such inclusions on the functional status of the microcapsules in vivo are unknown. Here we found that comparing the standard with the enhanced APA microcapsules; there was no difference in the rates of diffusion of recombinant products of different sizes, that is, human factor IX (FIX, 65 kDa), murine IgG (150 kDa), and a lysosomal enzyme, beta-glucuronidase (300 kDa), thus providing a key requirement of such an immunoprotective device. Furthermore, the creatine phosphokinase activity and myosin heavy chain staining (markers for differentiation of the myoblasts) and the cell number per capsule in the enhanced microcapsules indicated a higher degree of differentiation and proliferation when compared to the standard microcapsules, thus demonstrating an improved microenvironment for the encapsulated cells. Efficacy was tested in a melanoma cancer tumor model by treating tumor induced by B16-F0/neu tumor cells in mice with myoblasts secreting angiostatin from either the standard or enhanced APA microcapsules. Mice treated with enhanced APA-microcapsules had an 80% reduction in tumor volume at day 21 compared to a 70% reduction in those treated with standard APA-microcapsules. In conclusion, enhancement of APA microcapsules with growth factors and collagen did not adversely affect their permeability property and therapeutic efficacy. However, the enhanced differentiation and viability of the encapsulated myoblasts in vivo should be advantageous for long-term delivery with this method of gene therapy. PMID- 16470810 TI - Sphingosine-1-phosphate is released by cerebellar astrocytes in response to bFGF and induces astrocyte proliferation through Gi-protein-coupled receptors. AB - The mitogenic role of sphingosine-1-phosphate (S1P) and its involvement in basic fibroblast growth factor (bFGF)-induced proliferation were examined in primary cultures of cerebellar astrocytes. Exposure to bFGF resulted in a rapid increase of extracellular S1P formation, bFGF inducing astrocytes to release S1P, but not sphingosine kinase, in the extracellular milieu. The SK inhibitor N,N dimethylsphingosine inhibited S1P release as well as bFGF-induced growth stimulation. S1P application in quiescent astrocytes caused a dose-dependent increase in DNA synthesis. This gliotrophic effect was induced by a brief exposure to low nanomolar S1P, mimicked by the S1P receptor agonist dihydro-S1P, and inhibited by pertussis toxin (PTX), an inactivator of G(i)/G(o)-proteins. S1P also induced activation of extracellular signal-regulated kinase that was inhibited again by PTX. Moreover, the S1P lyase inhibitor 4-deoxypyridoxine induced the cellular accumulation of S1P but did not affect DNA synthesis. These results support the view that S1P exerted a mitogenic effect on cerebellar astrocytes extracellularly, most likely through cell surface S1P receptors. In agreement, mRNAs for S1P1, S1P2, and S1P3 receptors are expressed in cerebellar astrocytes (Anelli et al., 2005. J Neurochem 92:1204-1215). Ceramide, a negative regulator of astrocyte proliferation and down-regulated by bFGF (Riboni et al., 2002. Cerebellum 1:129-135), efficiently inhibited S1P-induced proliferation. The S1P action appears to be part of an autocrine/paracrine cascade stimulated by bFGF and, together with ceramide down-regulation, essential for astrocytes to respond to bFGF. The results suggest that S1P and bFGF/S1P may play an important role in physiopathological glial proliferation, such as brain development, reactive gliosis and brain tumor formation. PMID- 16470811 TI - Composite coating of bonelike apatite particles and collagen fibers on poly L lactic acid formed through an accelerated biomimetic coprecipitation process. AB - Collagen and apatite were coprecipitated as a composite coating on poly L-lactic acid (PLLA) in an accelerated biomimetic process. The incubation solution contained collagen (1 g/L) and simulated body fluid with 5 times inorganic ionic concentrations as human blood plasma. The coating formed on PLLA films and scaffolds after a 24-h incubation was characterized by using energy-dispersive X ray spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, and scanning electron microscopy (SEM). It was shown that the coating contained carbonated bonelike apatite and collagen, which was similar in composition to natural bone. SEM showed a complex composite coating of submicron bonelike apatite particulates combined with collagen fibrils. It is expected that such biocomposite coating may better facilitate cell interaction and osteoconductivity. This work provided an efficient process to obtain bonelike apatite/collagen composite coating, which is potentially useful in bone tissue engineering. PMID- 16470812 TI - TGF-beta1 immobilized tri-co-polymer for articular cartilage tissue engineering. AB - Tri-co-polymer with composition of gelatin, hyaluronic acid and chondroitin-6 sulfate has been used to mimic the cartilage extracellular matrix as scaffold for cartilage tissue engineering. In this study, we try to immobilize TGF-beta1 onto the surface of the tri-co-polymer sponge to suppress the undesired differentiation during the cartilage growth in vitro. The scaffold was synthesized with a pore size in a range of 300-500 microm. TGF-beta1 was immobilized on the surface of the tri-co-polymer scaffold with 1-ethyl-3-(3 dimethylaminopropyl) carbodiimide (EDC) as a crosslinking agent. Tri-co-polymer scaffolds with and without TGF-beta1 were seeded with porcine chondrocytes and cultured in a spinner flask for 2, 4, and 6 weeks. The chondrocytes were characterized by the methods of immunohistochemical staining with anti-type II collagen and anti-S-100 protein monoclonal antibody, and RT-PCR. After culturing for 4 weeks, chondrocytes showed positive in S-100 protein, Alcian blue, and type II collagen for the scaffold with TGF-beta1 immobilization. There is no observed type I and type X collagen expression in the scaffolds from the observation of RT PCR. In addition, the scaffold without TGF-beta1 immobilization, type X collagen, can be detected after cultured for 2 weeks. Type I collagen was progressively expressed after 4 weeks. These results can conclude that TGF-beta1 immobilized scaffold can suppress chondrocytes toward prehypertrophic chondrocytes and osteolineage cells. The tri-co-polymer sponge with TGF-beta1 immobilization should have a great potential in cartilage tissue engineering in the future. PMID- 16470813 TI - A mechanism on why slower polymerization of a dental composite produces lower contraction stress. AB - It has been well documented that the rate of polymerization of a dental composite often affects its polymerization contraction stress. In most cases, a slower cure produces a lower stress. To investigate the mechanism behind this, we prepared an unfilled dimethacrylate resin sample and photocured it using two light irradiances, both with the same total irradiation energy. We measured the polymerization-induced shrinkage from the unbonded surface of a class I restoration, contraction stress, extent of polymerization, and flexural modulus. The resin specimens cured under the two irradiances achieved the same extent of polymerization and developed an identical amount of shrinkage from the unbonded surface. But those cured under the lower irradiance possessed a lower contraction stress and a lower flexural modulus than those cured under the higher irradiance. We demonstrated that the stress level did not respond to the extent of viscous flow of the curing resin because the slower polymerization did not produce more shrinkage from the unbonded surface. Instead, the lower stress is likely due to a lower modulus of the cured resin. To explain why the cured resin with an identical extent of polymerization can have different moduli, we proposed that slower polymerization produces a higher level of structural inhomogeneity, which reduces the rigidity of the cured resin. PMID- 16470814 TI - Oxidation and oxidation potential in contemporary packaging for polyethylene total joint replacement components. AB - The packaging and chemical stability of both conventional and highly crosslinked polyethylene (PE) components available for clinical use in Italy were analyzed. A total of 100 sterilized PE components were entered by 18 orthopedic manufacturers into the study. Six of the manufacturers were Italian and the remaining were based in Europe or America. Hydroperoxide, oxidation, and trans-vinylene levels within the PE components were characterized using Fourier-transform infrared spectroscopy (FTIR). None of the 31 gas-sterilized components had detectable free radicals, hydroperoxide content, or oxidation. Among radiation-sterilized inserts, the highest oxidation and hydroperoxide levels were associated with gas permeable and polymer-barrier packaging. To the authors' knowledge, this is the first study that relates elevated oxidation and hydroperoxide content in gamma sterilized PE components to certain types of contemporary, polymeric barrier packaging. PMID- 16470815 TI - Critical comparison of two methods for the determination of nanomechanical properties of a material: application to synthetic and natural biomaterials. AB - Two methods used for determining the elastic modulus (E) and hardness (H) of a material--the original version of the well-known Oliver-Pharr Method, OOPM, and a variant of it called the Modified Slopes Method, MSM--were critically compared. The nanoindentation test results, of indenter load-versus-indenter displacement, were recorded for six series of specimens, three of commercially-available acrylic bone cements (Palacos R and Cemex XL) and three of bones (human, bovine, and mouse). In the first series, the specimens were prepared from Palacos R cement mantles retrieved from cemented total hip joint replacements after 11 months, 11 years, and 21 years in vivo. In the second and third series, the specimens were fabricated from hand- and vacuum-mixed dough of Cemex XL cement, respectively. In the fourth, fifth, and sixth series, the specimens were prepared from fresh frozen cortical bone of human tibia, plexiform bone from fresh bovine tibia, and femora from inbred mice, respectively. It was found that, for a given material, the values of E or H computed using OOPM and MSM are not significantly different. However, the recommendation is that MSM is preferable because it is straightforward-only the nanoindentation measurements and values of constants that depend on the geometry of the indenter used are needed. In contrast, when the OOPM is used, there is a critical input (the indenter tip area function), whose computation is problematic. The article also includes a succinct discussion of factors that affect the values of material properties computed from nanoindentation measurements, such as the loading rate and the surface roughness of the test specimen. PMID- 16470816 TI - Morphological assessment of the behavior of three-dimensional anionic collagen matrices in bone regeneration in rats. AB - The osteogenic behavior of three-dimensional collagenic and anionic matrices (M3DGA) implanted in critical bone defects of 8 mm diameter was morphologically assessed. The defects were performed in the calvaria of 48 adult male (Wistar) rats, and observed at days 15, 30, and 60. The animals were distributed in four groups of equal number: GA1 (M3DGA with 60 min of cross-linking in glutaraldehyde [GA]); GA2 (M3DGA with 30 min of cross-linking in GA); GA3 (M3DGA with 15 min of cross-linking in GA); and G4 (control group, without any implanted biomaterial). The M3DGAs were biocompatible, with mild and regressive chronic granulomatous inflammation, associated with the degradation of their fibers. Furthermore, tissue neovascularization and neomineralization was noted, with statistically significant differences in these amounts among the groups (GA1, GA2, and GA3) and G4. In G4 neoformation was limited to the edges of the defect as well as interstitial fibrosis. Under the experimental conditions and for the parameters analyzed, the M3DGAs had osteogenic behavior, which was more evident in M3DGA 15 min. PMID- 16470817 TI - Pulsatile dynamic stiffness of cartilage-like materials and use of agarose gels to validate mechanical methods and models. AB - Stiffness is a fundamental indicator of the functional state of articular cartilage. Reported test modes include compressive incremental strain to determine the equilibrium modulus, and sinusoidal strain to determine the dynamic modulus and stress/strain loss angle. Here, initial development is described for a method recognizing that gait is pulsatile. Agarose gels have been used by others for validation or comparison of mechanical test methods and models for cartilage and proteoglycan aggregate. Accordingly, gels ranging from 0.5 to 20% agarose were prepared. Pulsatile stiffness in both indentation and unconfined compression were closely reproducible. Stiffness as a function of agarose concentration rose exponentially, as found using other methods. Indentation stiffness was higher than for unconfined compression and ranged from approximately 2.0 kPa for 0.5% gel to approximately 3,800 kPa for 20% gel. Pulsatile dynamic stiffness appears to be a useful method, although further development is needed. Agarose gel stiffness values obtained by other methods were reviewed for comparison. Unfortunately, reported values for a given agarose concentration ranged widely (e.g. fourfold) even when test methods were similar. Causes appear to include differences in molecular weight and gel preparation time temperature regimens. Also, agarose is hygroscopic, leading to unintended variations in gel composition. Agarose gels are problematic materials for validation or comparison of cartilage mechanical test methods and models. PMID- 16470819 TI - Ab initio prediction of peptide-MHC binding geometry for diverse class I MHC allotypes. AB - Since determining the crystallographic structure of all peptide-MHC complexes is infeasible, an accurate prediction of the conformation is a critical computational problem. These models can be useful for determining binding energetics, predicting the structures of specific ternary complexes with T-cell receptors, and designing new molecules interacting with these complexes. The main difficulties are (1) adequate sampling of the large number of conformational degrees of freedom for the flexible peptide, (2) predicting subtle changes in the MHC interface geometry upon binding, and (3) building models for numerous MHC allotypes without known structures. Whereas previous studies have approached the sampling problem by dividing the conformational variables into different sets and predicting them separately, we have refined the Biased-Probability Monte Carlo docking protocol in internal coordinates to optimize a physical energy function for all peptide variables simultaneously. We also imitated the induced fit by docking into a more permissive smooth grid representation of the MHC followed by refinement and reranking using an all-atom MHC model. Our method was tested by a comparison of the results of cross-docking 14 peptides into HLA-A*0201 and 9 peptides into H-2K(b) as well as docking peptides into homology models for five different HLA allotypes with a comprehensive set of experimental structures. The surprisingly accurate prediction (0.75 A backbone RMSD) for cross-docking of a highly flexible decapeptide, dissimilar to the original bound peptide, as well as docking predictions using homology models for two allotypes with low average backbone RMSDs of less than 1.0 A illustrate the method's effectiveness. Finally, energy terms calculated using the predicted structures were combined with supervised learning on a large data set to classify peptides as either HLA-A*0201 binders or nonbinders. In contrast with sequence-based prediction methods, this model was also able to predict the binding affinity for peptides to a different MHC allotype (H-2K(b)), not used for training, with comparable prediction accuracy. PMID- 16470820 TI - Megestrol attenuates the hormonal response to CCK-4-induced panic attacks. AB - Progestational hormones may have anxiolytic properties. CCK-4 (cholecystokinin tetrapeptide) can be used pharmacologically to induce panic attacks both in normal controls and patients suffering from panic disorder. In this study we compared the effects of pretreatment with the progestational hormone megestrol and placebo on CCK-4-induced panic attacks and stress hormone release in healthy male controls. Using a double-blind balanced design, we pretreated 10 medically and psychiatrically healthy male controls with placebo or megestrol 160 mg at 11 p.m. and 8 a.m. (sigma=320 mg) prior to the experiment. Following 1 h of rest, 12 blood samples were drawn between 1,000 h and 1,300 h and analyzed for ACTH and cortisol levels. At 1,100 h, subjects received an intravenous injection of 50 microg CCK-4. Clinical ratings were performed at 1,045 h and 1,110 h, and included the Acute Panic Inventory (API), International Diagnostic Checklist (IDCL), as well as a visual analog scale (VAS) for anxiety and tension. CCK-4 significantly increased anxiety and tension. Pretreatment with megestrol showed no significant effect on clinical ratings. Baseline ACTH and cortisol levels, as well as ACTH and cortisol levels after administration of CCK-4, were significantly reduced after pretreatment with megestrol. In a sample of healthy male controls, pretreatment with megestrol had a profound effect on the hypothalamic-pituitary-adrenal (HPA) axis, whereas the clinical effects on panic attacks were weak. Further studies in a larger sample of subjects, including both females and patients suffering from panic disorder, seem warranted. PMID- 16470821 TI - Principal component analysis and artificial neural network analysis of oral tissue fluorescence spectra: classification of normal premalignant and malignant pathological conditions. AB - Pulsed laser-induced autofluorescence spectroscopic studies of pathologically certified normal, premalignant, and malignant oral tissues were carried out at 325 nm excitation. The spectral analysis and classification for discrimination among normal, premalignant, and malignant conditions were performed using principal component analysis (PCA) and artificial neural network (ANN) separately on the same set of spectral data. In case of PCA, spectral residuals, Mahalanobis distance, and scores of factors were used for discrimination among normal, premalignant, and malignant cases. In ANN, parameters like mean, spectral residual, standard deviation, and total energy were used to train the network. The ANN used in this study is a classical multiplayer feed-forward type with a back-propagation algorithm for the training of the network. The specificity and sensitivity were determined in both classification schemes. In the case of PCA, they are 100 and 92.9%, respectively, whereas for ANN they are 100 and 96.5% for the data set considered. PMID- 16470822 TI - PLA stereocopolymers as sources of bioresorbable stents: preliminary investigation in rabbit. AB - The aim of the present work was to evaluate whether the degradation of PLA-based bioresorbable stents can be modulated via the configuration of repeating units as it is the case in other applications like osteosynthesis. The first obstacle was finding a stent design that could allow implantation in the aorta of a rabbit taken as a model of a human coronary artery. In the absence of guidelines other than those tentatively proposed in patents, several simple designs were considered that allowed us to evaluate the fate of the stents made of poly(lactic acid) stereocopolymers with L/(L + D) ratio of 0.92 (PLA92) and 0.50 (PLA50) up to 6 months post in vivo implantation. Our findings show the feasibility of bioresorbable stenting using PLA stereocopolymers and that PLA50 degraded faster than PLA92. Therefore, using stereocopolymers appears as a means to vary the degradation rate and adapt it to the artery remodelling process that is very much dependent on the release of the stenting stress protection. PMID- 16470824 TI - Three-dimensional printing of porous polyethylene structure using water-based binders. AB - This study demonstrated a freeform fabrication of porous hydrophobic polyethylene parts by three-dimensional printing technique, using water-based binder. This involved initially making a green structure by printing a binder onto the mixture of polyethylene and water soluble adhesive powder bed. The green structure was thermal treated to melt polyethylene fraction to achieve a well-connected structure within the part. Adhesive components were then leached out in water, leaving only porous structure of polyethylene. Specimens with ratios of polyethylene to adhesive from 80:20 to 20:80 were fabricated and characterized. It was found that increasing polyethylene content increased density, modulus, and strength while decreased porosity. Preliminary in vitro toxicity test of porous parts using L929 cells showed that the cells that were in contact with samples were healthy. No inhibition zone was observed. PMID- 16470823 TI - Cytotoxic effects of dental desensitizers on human gingival fibroblasts. AB - The purpose of this study was to evaluate the effects of three different desensitizers on the cell viability and morphology of human gingival fibroblasts (HGF). Human gingival tissues were obtained from individuals who have clinically, healthy periodontium. HGF were grown at 37 degrees C in humidified atmosphere of 5% CO2 in Dulbecco's modified eagle's medium, supplemented with glutamine, penicillin, streptomycin, and 10% fetal bovine serum. The cells were treated with different concentrations (0.1, 0.3, and 0.5 microL/mL) of desensitizers (Gluma Desensitizer, Seal&Protect, and MicroPrime). After 24- and 48-h exposure to the desensitizer solutions, the viable cells were examined using a hemocytometer. To monitor HGF viability, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetric assay was used and cell morphology was also observed at 48 h. Following exposure to concentrations of 0.1 microL/mL of test materials for 24 h, cell survival rates for Gluma Desensitizer (106%) and Micro Prime (62%) were not significantly different from the control, while it was significant for Seal&Protect (50%). Growing cells were significantly inhibited by all tested materials for 48 h (p < 0.05) in survival rates of 51, 47, and 31%, respectively. On the basis of the MTT assay, the cytotoxic effect of MicroPrime was more prominent, especially at high concentrations, than does Gluma Desensitizer and Seal&Protect. After exposure to Seal&Protect and MicroPrime, HGF became retracted, rounded in appearance and had loss of normal organization, leading to enlargement of intercellular space when compared with Gluma Desensitizer. As a conclusion, taking the limitations of an in vitro experiment into consideration, the cytotoxic effects were varied, depending on the chemical composition and exposure periods of the tested desensitizers. PMID- 16470825 TI - The nature of platinum in silicones for biomedical and healthcare use. AB - Silicone is an important biomaterial in many different biomedical and healthcare applications. Network formation in one type of silicone relies upon a chemical crosslinking reaction that typically employs a platinum catalyst. As a consequence, low concentrations of platinum may remain in certain medical devices designed for human use. The characteristics of platinum in silicone before, during, and after the crosslinking reaction have been well described in the literature. This review summarizes the relevant literature on the organometallic and analytical chemistry of platinum in silicone and thus provides a foundation for understanding the effects this platinum may have, if any, in the various biomedical and healthcare applications where it may be present. PMID- 16470826 TI - Relationship between fracture toughness and flexural strength in dental porcelains. AB - The aim of this study was to investigate the correlation between fracture toughness (K(Ic)) and flexural strength (FS) in dental porcelains. Porcelains with different leucite contents and clinical indications were used (A, B, C, D, and E). K(Ic) was determined by surface crack in flexure method (SCF) and FS was determined by four-point-bending test. Microstructural characterization was also carried out. The leucite contents of porcelains A, B, C, D, and E were, respectively, 22, 22, 6, 15, and 0%. Materials with higher leucite content (A and B) presented significantly higher K(Ic) values compared to materials with lower leucite content (C and E). The Weibull moduli (m) of porcelains A and B were statistically higher than those of the other three materials. Regarding characteristic strength (sigma(0)), porcelains D and E showed similar values and statistically higher than those of the other materials which were statistically different from each other. According to the regression analysis, sigma(0) increased with the increase of K(Ic) until approximately 0.75 MPa m(1/2). After that, the increase in K(Ic) was accompanied by a decrease in sigma(0). However, the Weibull modulus increased with the increase in K(Ic), especially for values greater than 0.80 MPa m(1/2). PMID- 16470827 TI - Synthesis and performance of amphiphilic copolymers for blood cell separation. AB - Three types of amphiphilic copolymers using n-butylmethacrylate (BMA) as a hydrophobic monomer, and each of N,N'-dimethylacrylamide (DMA), N acryloylmorpholine (AMO), and N-vinylpyrrolidone (VP) as hydrophilic comonomers were synthesized for coating filters used to remove leukocytes. The influence of the amphiphilic property of the resulting filters, which were composed of nonwoven fabrics coated with the above copolymers, on leukocyte removal and platelet permeation through the filters from whole blood was investigated. The platelet permeation ratio through hydrophobic noncoated filters was only 0.2%, because platelets in whole blood adhered easily to the hydrophobic filter material. However, filters coated with poly(AMO-co-BMA) of high AMO content showed a much higher platelet permeation ratio (nearly 90%). Further, the filters coated with poly(DMA-co-BMA) also showed high permeation ratios of platelets (more than 78%) over a broad range of DMA content in the copolymer. On the other hand, the coated filters showed slightly a higher permeation ratio of leukocytes than did the noncoated filters, resulting from the increase in hydrophilicity of the surface of the filters. Moreover, the coating of the amphiphilic copolymers on the surface of the nonwoven fabrics may have affected the pore size of the filters, affecting the permeation ratio of leukocytes more strongly than that of platelets. The coated filters effectively improved platelet permeation through the filters, with a slight increase in the permeation ratio of leukocytes. PMID- 16470828 TI - Multivariate analysis of surface physico-chemical properties controlling biofilm formation on orthodontic adhesives prior to and after fluoride and chlorhexidine treatment. AB - Biofilm formation on orthodontic adhesives is a serious clinical problem, as it leads to enamel demineralization around fixed orthodontic appliances, often leaving white spot lesions after their removal. The aim of this work was to determine the influence of surface physico-chemical properties of four commonly used orthodontic adhesives (Concise, Fuji ORTHO LC, Ketac Cem mu, and Transbond XT) on early bacterial biofilm formation. In addition, effects of two commercially available mouthrinses (0.05% sodium fluoride and 0.2% chlorhexidine gluconate) on these properties and biofilm formation were determined. Water contact angles on the adhesives decreased after fluoride and chlorhexidine treatment, concurrent with an increase in carbon and a decrease in oxygen surface concentrations, except for Transbond, as determined by X-ray photoelectron spectroscopy. No fluorine was detected on any of the adhesive surfaces after fluoride treatment, while all surfaces showed chlorine after chlorhexidine treatment. Surface roughness of the adhesives measured using three-dimensional optical profilometry was around 4 microm and found not to be a factor governing early biofilm formation. Multiple linear regression analysis indicated that early biofilm formation by Streptococcus sanguis could be explained in a model comprising hydrophobicity and the prevalence of oxygen- and nitrogen-rich components on the adhesive surfaces. PMID- 16470829 TI - Biodegradable polymer-silica xerogel composite microspheres for controlled release of gentamicin. AB - Single and double layered composite microspheres were prepared by encapsulating gentamicin-loaded silica xerogels with biodegradable PLGA polymers (poly(DL lactide-co-glycolide)). The in vitro drug release properties of both the composite microspheres were investigated. The single layered composite microspheres showed a high initial burst, followed by two sustained release stages lasting for approximately 6 weeks. The two sustained release stages of the single layered composite microspheres could be attributed to the swelling and bulk erosion of the polymer encapsulations, respectively. In comparison with the single layered composite microspheres, the double layered composite microspheres realized a much reduced initial burst together with three sustained release stages. The whole release period of the double layered composite microspheres could last more than 9 weeks. These distinct behaviors make the double layered composite microspheres promising as a new drug release material for localized drug delivery applications. PMID- 16470830 TI - Effect of pH and temperature on orthodontic NiTi wires immersed in acidic fluoride solution. AB - Orthodontic arch wires can confront diverse pH and temperature conditions in the oral cavity after replacement. The current study evaluated the effect of pH and temperature on orthodontic NiTi arch wires after immersion in an acidic fluoride solution. The acetic acid added for the adjustment of pH forms hydrofluoric acid (HF). More HF was formed in a solution of lower pH with higher temperature than that of the higher pH with lower temperature. Within the same pH value, the temperature of the solution affected the concentration of HF. Each as-received wire exhibited quite different microhardness values. The reduction of microhardness, 1.2-5.7%, occurred after immersion. Within the same product, however, the pH and temperature had minor influence on the reduction. The volumetric weight change, concentration of the released elements, and surface morphology were influenced by pH and temperature. At pH 3.5 of 60 degrees C solution, the greatest weight loss, release of elements, and corrosion of surface occurred from the wires. At pH 6, on the other hand, no such loss or release occurred regardless of temperature. At 5 degrees C solution, the surface exhibited minor corrosion regardless of pH value. PMID- 16470831 TI - The effect of fiber insertion on fracture resistance of endodontically treated molars with MOD cavity and reattached fractured lingual cusps. AB - In this study, the effect of flowable composite reinforced with a leno wave ultra high modulus (LWUHM) polyethylene fiber (Ribbond) on fracture resistance of endodontically treated molars with MOD cavity and lingual cuspal fracture was evaluated. Sixty sound extracted human mandibular molars were randomly assigned to six groups (n = 10). Group 1 served as control. Teeth in groups 2-6 received root canal treatment and a MOD cavity preparation. Teeth in group 2 were kept unrestored. Lingual walls of specimens in groups 3-6 were fractured at the CEJ and reattached (C&B Super-Bond). Group 3 was kept unrestored, and group 4 was restored with a composite resin (CR) (AP-X). In group 5, a flowable resin (FR, Protect Liner F) and in group 6, a Ribbond in combination with FR were inserted inside the cavity before CR restoration. After finishing and polishing, the specimens were subjected to compressive loading perpendicular to the occlusal surface at a crosshead speed of 1 mm/min. The mean load necessary to fracture were recorded in Newton and the results were statistically analyzed. MOD cavity preparation reduced fracture resistance of endodontically treated teeth (p < 0.05). Fracture resistance of rebonded fractured specimens was found to be similar to that of the nonfractured samples (p > 0.05). Use of LWUHM polyethylene fiber Ribbond increased fracture strength of endodontically treated molar teeth with MOD cavity preparation and cuspal fracture (p < 0.05). As a result, it was concluded that the insertion of Ribbond inside the cavity has a positive effect on fracture strength of endodontically treated molar teeth with MOD cavity preparation and cuspal fracture. PMID- 16470832 TI - Transverse fracture of brittle bilayers: relevance to failure of all-ceramic dental crowns. AB - This study examines the behavior of cracks approaching interfaces in all-ceramic dental crown-like bilayers. Flat specimens are fabricated by fusing porcelain veneers onto yttria-tetragonal-zirconia-polycrystal (Y-TZP) and alumina core ceramic plates, with veneer/core matching to minimize residual thermal expansion mismatch stresses. Vickers indentations are placed on either side of the interfaces, at systematically decreasing distances, so that the lead corner cracks approach and intersect the interfaces in a normal orientation. Cracks originating in the porcelain arrest at the boundaries and, after further diminution in indentation distance, deflect along the interface without penetration into the tough core ceramic. Cracks initiating in the core ceramic pass unimpeded into the weaker porcelain without deflection, and with abrupt increase in crack size. These latter cracks, because of their lack of containment within the core layer, are regarded as especially dangerous. Implications concerning the design of optimal dental crowns in relation to materials optimization are considered. PMID- 16470833 TI - The internalized CdSe/ZnS quantum dots impair the chondrogenesis of bone marrow mesenchymal stem cells. AB - Mesenchymal stem cells (MSCs) are capable of differentiating into multiple cell lineages and are useful for therapeutic applications. Labeling the MSCs with fluorescent probes is beneficial in tracing the fate of MSCs after implantation. We have introduced the CdSe/ZnS quantum dots (QDs) into the human bone marrow MSCs and examined the effects of QDs on the proliferation and chondrogenesis of the cells. The internalized QDs were found localized in perinuclear regions and remained there after a number of cell passages. The presence of QDs did not affect the proliferation of cells or the size of chondrospheres formed, when subjected to chondrogenesis induction. However, the expression of mRNA and protein of type II collagen and aggrecan in the chondrospheres was significantly inhibited in cells labeled with QDs, suggesting impaired chondrogenesis. Our results that the presence of QDs interferes with the chondrogenic differentiation of MSCs raise concerns in using the QDs as fluorescence tracers for stem cells. PMID- 16470834 TI - Behavior of human osteoblast-like cells in contact with electrodeposited calcium phosphate coatings. AB - Calcium-deficient hydroxyapatite (Ca-def-HAP) thin films were elaborated on Ti6Al4V substrates by electrodeposition. The coatings exhibit two different morphologies and crystallinities. Human osteoblast-like cells (MG-63) were cultured on the surfaces of these materials; the cell content and viability were evaluated up to 28 days. The scanning electron microscopy and biological investigations showed cells with a normal morphology, good proliferation, and viability from 7 to 21 days. But after 28 days, the number of live cells decreases in both cases; however, this decrease is less important in the case of calcium phosphate (CaP) coating surface when compared with the control (cell culture plastic). The cells cultured on Ca-def-HAP coating exhibit more cellular extensions and extracellular matrix. RT-PCR for type I collagen, alkaline phosphatase, and osteocalcin studies were also carried out, and was found that the CaP enhances gene expression of ALP and OC and thus the differentiation of osteoblast-like cells. Moreover, this study shows that the difference in the morphology of CaP coatings has no effect on the biocompatibility. PMID- 16470836 TI - The VMFCI method: a flexible tool for solving the molecular vibration problem. AB - The present article introduces a general variational scheme to find approximate solutions of the spectral problem for the molecular vibration Hamiltonian. It is called the "vibrational mean field configuration interaction" (VMFCI) method, and consists in performing vibrational configuration interactions (VCI) for selected modes in the mean field of the others. The same partition of modes can be iterated until self-consistency, generalizing the vibrational self-consistent field (VSCF) method. As in contracted-mode methods, a hierarchy of partitions can be built to ultimately contract all the modes together. So, the VMFCI method extends the traditional variational approaches and can be included in existing vibrational codes based on the latter approaches. The flexibility and efficiency of this new method are demonstrated on several molecules of atmospheric interest. PMID- 16470835 TI - FGF ligand family mRNA expression profile for mouse preimplantation embryos, early gestation human placenta, and mouse trophoblast stem cells. AB - Signaling by fibroblast growth factor (FGF) is essential is for trophoblast stem (TS) cells and preimplantation embryos. FGF4 provides essential signaling, but the expression of the complete set of 23 FGF family members has not been analyzed. Here, semi-quantitative RT-PCR and microarray analyses were used to define expression of all FGF ligand mRNA. RT-PCR was done for developmentally important FGF subfamilies, FGF10/FGF22 and FGF8/FGF17/FGF18 as well as FGF11. FGF4 and FGF18 are detected at highest levels by RT-PCR and microarrays. FGF10 was detected at low levels in both assays. FGF11 was detected at moderate levels by microarray, but not by RT-PCR. FGF17 was detected at low levels by array and moderate levels by RT-PCR. FGF8 and FGF22 were detected by RT-PCR, but not by microarrays during late cleavage divisions. FGF8, FGF5, and FGF9 were detected in the oocyte by microarray. FGF2, FGF3, and FGF7 were not detected by RT-PCR or microarrays and FGF13, FGF14, and FGF23 were not detected by microarray. Since a major role of FGF is to maintain TS cells, we tested human and mouse placental cell lines and early gestation human placenta for expression of FGF ligands. Expression in mouse TS cells was compared with preimplantation embryos, and human placental cell line expression was compared with human placenta, to infer which ligands are expressed in placental lineage vs. other cell lineages. The data suggest that human and mouse placenta share FGF18 and its high expression suggests preimplantation and early placental function. PMID- 16470837 TI - Fine structural dependence of ultraviolet reflections in the King Penguin beak horn. AB - The visual perception of many birds extends into the near-ultraviolet (UV) spectrum and ultraviolet is used by some to communicate. The beak horn of the King Penguin (Aptenodytes patagonicus) intensely reflects in the ultraviolet and this appears to be implicated in partner choice. In a preliminary study, we recently demonstrated that this ultraviolet reflectance has a structural basis, resulting from crystal-like photonic structures, capable of reflecting in the near-UV. The present study attempted to define the origin of the photonic elements that produce the UV reflectance and to better understand how the UV signal is optimized by their fine structure. Using light and electron microscopic analysis combined with new spectrophotometric data, we describe here in detail the fine structure of the entire King Penguin beak horn in addition to that of its photonic crystals. The data obtained reveal a one-dimensional structural periodicity within this tissue and demonstrate a direct relationship between its fine structure and its function. In addition, they suggest how the photonic structures are produced and how they are stabilized. The measured lattice dimensions of the photonic crystals, together with morphological data on its composition, permit predictions of the wavelength of reflected light. These correlate well with experimentally observed values. The way the UV signal is optimized by the fine structure of the beak tissue is discussed with regard to its putative biological role. PMID- 16470838 TI - Tumor selective replication of Newcastle disease virus: association with defects of tumor cells in antiviral defence. AB - To investigate tumor-selective viral replication, we compared several tumorigenic human cell lines to nontumorigenic human cells from the blood for the sensitivity to become infected by a recombinant lentogenic strain of Newcastle Disease Virus (NDV) with incorporated transgene EGFP (NDFL-EGFP). Although fluorescence signals in nontumorigenic cells were only weak or missing completely, a massive and long lasting transgene-expression was observed in all tumor cell lines. The majority of tumor cells (50-95%) could be infected, and viral replication was associated with an increase in the cell surface density of viral antigens. To clarify the underlying mechanism of the observed difference in virus susceptibility we examined the kinetics of interferon-induced antiviral enzymes because NDV is a strong type-I interferon inducer. This analysis revealed several defects of tumor cells in their antiviral defence responses: They showed no response to UV inactivated NDV, whereas nontumorigenic cells reacted with induction of high levels of the antiviral enzymes PKR and MxA. Upon coincubation with live NDV, tumor cells showed a delayed response in the increased expression of the antiviral enzymes in comparison with PBMC. In nontumorigenic cells the replication cycle of NDV stopped after the production of positive-strand RNA, while tumor cells continued in the replication cycle and copied viral genomes 10 50 hr after infection. These results can explain the tumor selective replication behavior of this interesting antineoplastic virus. PMID- 16470839 TI - A long-term follow-up study on the natural course of snus-induced lesions among Swedish snus users. AB - Snus-induced lesions (SILs) are mucosal changes that are regularly seen in users of moist snuff (snus). Their role in oral carcinogenesis remains undefined. Our aim was to assess the natural course of SILs over several decades. A cohort of 1,115 individuals with SILs, confirmed in 1973-1974 during a population-based survey was followed for 27-29 years through multiple record linkages with virtually complete population- and health registers. A sample (n = 267) of the cohort members were invited for reexamination after 19-22 years. Register-based follow-up through January 2002 revealed a total of 3 incident cases of oral cancer (standardized incidence ratio of 2.3, 95% CI 0.5-6.7), none of which occurred at the site of the original SIL. There was a strong association noted between the degree of SIL and current snus consumption. The SILs had disappeared in all 62 individuals who had permanently quit using snus. In no case did we observe an important clinical change for the worse among individuals who had decreased their use or continued unabatedly. While the incidence of oral cancer in this cohort of individuals with SILs tended to be higher than expected, we conclude that cancers rarely occur at the site of lesions observed in the distant past. PMID- 16470840 TI - Iressa induces cytostasis and augments Fas-mediated apoptosis in acinic cell adenocarcinoma overexpressing HER2/neu. AB - Understanding the role of signal transduction in regulating pathways responsible for cell growth, survival and apoptosis is critical for cancer therapy. We developed and characterized a HER2/neu and Fas overexpressing cell line (BNT.888 ACA2) from a salivary gland adenocarcinoma that arose in a HER2/neu transgenic mouse. We evaluated the effects of Iressa on signal transduction networks downstream of the activated HER2 and the impact on proliferation, cell cycle and apoptosis. Iressa treatment diminished phosphorylation of the HER2/neu and EGFR. Phosphorylation of STAT-3 also decreased and mitogenic signaling through the MAPK pathways was greatly reduced. Cyclin D1 levels decreased, and cells were arrested in G0 and failed to enter S-phase because of hypophosphorylation of Rb and to traverse the G2M checkpoint because of degradation of cyclin B1. Cytostasis occurred within 48 hr at 250-500 nM Iressa. Levels of proapoptotic factors (bim and bax) increased and levels of antiapoptotic factors (bcl-2 and bcl-xL) decreased in a dose-dependent manner. Higher doses of Iressa diminished phosphorylation of Akt slightly, but failed to induce apoptosis. Fas antibody was a potent agonist of apoptosis. Pretreatment with Iressa (1 microM, 24 hr) greatly enhanced Fas-mediated apoptosis as determined by Annexin V binding, cleavage of caspase-3 and PARP. Augmentation of apoptosis was associated with increased Fas expression and membrane localization. Iressa pretreatment increased bid activation, cleavage of caspases -3, -9 and -12 and stress signaling via c Jun. These data showing that Iressa induces cytostasis and primes the extrinsic (Fas) and intrinsic (mitochondrial and endoplasmic reticulum) apoptotic pathways should lead to the development of novel therapeutic targets and strategies. PMID- 16470841 TI - What determines the spectrum of protein native state structures? AB - We present a brief summary of the key factors underlying protein structure, as developed in the investigations of Pauling, Ramachandran, and Rose. We then outline a simplified physical model of proteins that focuses on geometry and symmetry. Although this model superficially appears unrelated to the detailed chemical descriptions commonly applied to proteins, we show that it captures the essential elements of the chemistry and provides a unified framework for understanding the common characteristics of folded proteins. We suggest that the spectrum of protein native state structures is determined by geometry and symmetry and the role of the sequence is to choose its native state structure from this predetermined menu. PMID- 16470842 TI - Vascular perfusion of human lung cancer in a rat orthotopic model using dynamic contrast-enhanced magnetic resonance imaging. AB - Lung cancer is the leading cause of death among cancers. Early detection and diagnosis present a major goal in the efforts to improve survival rates of lung cancer patients. Changes in angiogenic activity and microvascular perfusion properties in cancers can serve as markers of malignancy. The aim of this study was to employ MRI means to measure the microvascular perfusion parameters of orthotopic nonsmall cell lung cancer, using the experimental rat model. Anatomical and dynamic contrast-enhanced lung images were acquired at high spatial resolution, and registered and analyzed, pixel by pixel and globally, by means of a model-based algorithm. The MRI output yielded color-coded parametric images of the influx and efflux transcapillary transfer constants that indicated rapid microvascular perfusion. The transfer constants were about 1 order of magnitude higher than those found in other tumors or in nonorthotopic lung cancer, with the influx constant median value of 0.42 min(-1) and the efflux constant median value of 1.61 min(-1). The rapid perfusion was in accord with the immunostaining of the capillaries, which suggested the tumor exploitation of the existing alveolar vessels. The results showed that high resolution, dynamic, contrast-enhanced MRI is an effective tool for the quantitative measurement of spatial and temporal changes in lung cancer perfusion and vasculature. PMID- 16470843 TI - Cyclic AMP synergistically enhances neuregulin-dependent ERK and Akt activation and cell cycle progression in Schwann cells. AB - The elevation of intracellular cAMP synergistically enhances the neuregulin dependent proliferation of cultured Schwann cells (SCs); however, the mechanism by which this occurs has not been completely defined. To better understand this mechanism, we investigated the effect of cAMP on the activation of the extracellular signal-regulated kinase (ERK) and phosphoinositide 3-kinase (PI3-K) Akt (PKB) pathways by heregulin, a member of the neuregulin family. Using primary cultures of adult SCs, we demonstrated that the adenylyl cyclase activator, forskolin, enhanced heregulin-dependent SC proliferation by reducing the time required for S-phase entry. When cAMP levels were increased, using either forskolin or a cell permeable analogue of cAMP, the heregulin-induced phosphorylation of ERK was converted from transient to sustained and the heregulin-induced phosphorylation of Akt was synergistically increased. Consistent with these observations, studies in which inhibitors of MEK, the upstream stimulating ERK kinase, and PI3-K were administered at different times following the onset of stimulation indicated that sustained high levels of both MEK/ERK and PI3-K/Akt activity before S-phase initiation were essential for S phase entry. Overall, these novel results indicate that in neuregulin-stimulated SCs the activation of cAMP-mediated pathways accelerates G1-S progression by prolonging ERK activation and concurrently enhancing Akt activation. PMID- 16470844 TI - Pulsed dye laser treatment of genital warts. AB - BACKGROUND AND OBJECTIVES: Genital warts represent benign epithelial proliferations induced by human papillomavirus. The goal of treatment is the clearance of visible warts. Different regimens are available. Flashlamp-pumped pulsed dye laser (FPDL) represents one of many treatment options for the management of viral warts (verrucae vulgares), its effectiveness being comparable with that of conventional therapies. We evaluated the effectivity of FPDL light for the treatment of genital warts. STUDY DESIGN/MATERIALS AND METHODS: A prospective study was performed to examine the efficacy of FPDL in untreated genital warts in which 22 patients were included. RESULTS: All patients showed complete remission after 1.59 (1-5) laser sessions and no scarring was observed. CONCLUSIONS: This study demonstrate that FPDL is a simple and safe, cost and time saving alternative treatment option for genital warts and should be listed in genital warts treatment guidelines. PMID- 16470846 TI - Thorium X: not yet a closed chapter. PMID- 16470845 TI - Photodynamic therapy with topical 5-aminolevulinic acid as a post-operative adjuvant therapy for an incompletely resected primary nasopharyngeal papillary adenocarcinoma: a case report. AB - BACKGROUND AND OBJECTIVES: Surgical excision of primary nasopharyngeal papillary adenocarcinoma is sometimes incomplete with remaining microscopic disease. Post operative radiotherapy only has limited efficacy but may cause many complications. STUDY DESIGN/MATERIALS AND METHODS: Photodynamic therapy (PDT) was used as a post-operative adjuvant therapy for an incompletely resected primary nasopharyngeal papillary adenocarcinoma. A special form of 20% topical 5 aminolevulinic acid (5-ALA), which was originally a liquid form and became a gel form after applied on the nasopharynx, was used as the photosensitizer. A 2-mm optic fiber delivered the light (633 nm wavelength) to the lesion with a fluence rate of 100 mW/cm2 generated by a diode laser under 5 mm 0 degrees endoscope assistance. The total energy delivered was 150 joules/cm2. RESULTS: No significant acute side effect was noted and the nasopharyngeal wound healed rapidly. The patient is alive without locoregional recurrence or distant metastasis for 5 years. Articulation, salivation, and swallowing functions are all well preserved. CONCLUSION: Post-operative adjuvant PDT can successfully cure an otherwise difficult to treat disease with preservation of good life quality of the patient. Potential complications of PDT (e.g., photosensitivity) can be prevented by a special formulation of topical 5-ALA preparation. PMID- 16470847 TI - Laser-induced bubbles in living cells. AB - BACKGROUND AND OBJECTIVES: Laser-activated micro- and nano-bubbles (LAB) in cells may be used as universal and sensitive non-toxic probes for measuring functional properties of individual cells. STUDY DESIGN/MATERIALS AND METHODS: Such bubbles can be detected and imaged by microscopy and flow cytometry. LABs in living blood and tumor cells were induced by pulsed (532 nm, 10 nanoseconds) laser radiation and detected by the thermal lens optical method. RESULTS: Registered lifetime and maximal diameter of the studied LABs varied within the ranges of 0.02-10 microseconds and 0.44-100 microm, respectively. LAB parameters, thresholds and probabilities, were found to depend upon the physiological state of cells. Specificity and sensitivity of LAB cytometry were increased due to the use of light-absorbing nanoparticles conjugated to specific monoclonal antibodies. CONCLUSIONS: LAB were found to be the universal phenomena that can be used for sensitive and non-invasive monitoring of any individual cell, intact or nanoparticle-treated. PMID- 16470849 TI - Novel conserved hydrolase domain in the CLCA family of alleged calcium-activated chloride channels. AB - Advanced protein structure prediction methods combined with structure modeling show that the mammalian proteins, described until now as calcium-activated chloride channels (CLCAs), appear in fact to be membrane anchored metal-dependent hydrolases, possibly proteases. A metallohydrolase structural domain was predicted, unexpectedly, in the CLCA sequences. The well-conserved active site in the modeled structure of this hydrolase domain allows the prediction of catalytic action similar to that of metalloproteases. A number of protein structure prediction methods suggest the overall fold of the N-terminal hydrolase domain to be most similar to that of zinc metalloproteases (zincins), notably matrixins. This is confirmed by analysis of the three-dimensional structure model of the predicted CLCA1 hydrolase domain built using the known structure of the MMP-11 catalytic domain. Fragments of CLCA1 corresponding to the modeled hydrolase domain were expressed in Escherichia coli, and the resulting proteins were readily refolded into monomeric soluble protein, indicating formation of stable independent domains. The homology model was used to predict putative substrate sequences. Homologs of mammalian CLCA genes were detected in the genomes of a vast array of multicellular animals: lower vertebrates, tunicates, insects, crustaceans, echinoderms, and flatworms. The hydrolase prediction is discussed in the context of published experimentally determined effects of CLCA proteins on chloride conductance. Altered proteolytic processing of full-length CLCA1 containing a mutation abolishing the predicted hydrolase activity is shown as initial experimental evidence for a role of the hydrolase domain in processing of mature full-length CLCA1. The hydrolase prediction together with the presented experimental data add to doubts about the function of CLCAs as chloride channels and strengthen the hypothesis of channel-activating and/or channel-accessory roles. PMID- 16470848 TI - Treatment effects of combined radio-frequency current and a 900 nm diode laser on leg blood vessels. AB - BACKGROUND AND OBJECTIVES: Effective laser treatment of leg veins remains a major challenge. The present study examined the safety and efficacy of a new technology for leg vein treatment combining 900 nm diode laser with radiofrequency (RF) current. STUDY DESIGN/MATERIALS AND METHODS: Forty patients, skin types II-IV, received a maximum of three treatments on 1-4 mm leg veins at 2-week intervals with a 900 nm diode laser (250 millisecond exposure time, average fluence 60 J/cm2) and RF (energy 100 J/cm3). Results were assessed after each treatment and at 2 and 6 months after the final session. Patients rated their satisfaction with the clinical outcome on a five-item scale. Clinician and computer analysis of the clinical photography was also performed, in addition to histological assessment. RESULTS: One or two sessions were required in the majority of patients. Shortly after treatment, histology revealed contracted vessels with perivascular edema. Side effects were extremely rare. The clinician 2- and 6-month assessments showed that 70% and 82.5% of subjects, respectively, achieved over 50% clearance, with patient and computer assessments lower and slightly higher, respectively. Treatments showed greater efficacy on thicker vessels and in the darker skin types. CONCLUSIONS: The success of the treatment, minimal side effects, and patient comfort suggest that this combination is an effective, safe technique for leg vein treatment. When compared to previous studies using diode laser alone, the very low fluence needed to achieve vessel clearance emphasizes the role of RF energy. PMID- 16470851 TI - Structural variation and immune recognition of the P1.2 subtype meningococcal antigen. AB - Neisseria meningitidis is a globally important cause of bacterial meningitis and septicemia. No comprehensive antimeningococcal vaccine is available, largely as a consequence of the high sequence diversity of those surface proteins that could function as components of a vaccine. One such component is the protein PorA, a major surface porin of this Gram-negative organism that has been used in a number of experimental and licensed vaccines. Here we describe a series of experiments designed to investigate the consequences for antibody recognition of sequence diversity within a PorA antigen. The binding of a 14-residue peptide, corresponding to the P1.2 subtype antigen, to the MN16C13F4 monoclonal antibody was sensitive to mutation of five out of the six residues within the epitope sequence. The crystal structure of the antibody Fab fragment, determined in complex with the peptide antigen, shows a remarkably hydrophobic binding site and interactions between the antigen and antibody are dominated by apolar residues. Nine intrachain hydrogen bonds are formed within the antigen which maintain the beta-hairpin conformation of the peptide. These hydrogen bonds involve residues that are highly conserved amongst different P1.2 sequence variants, suggesting that some positions may be conserved for structural reasons in these highly polymorphic regions. The sensitivity of antibody recognition of the antigen towards mutation provides a structural explanation for the widespread sequence variation seen in different PorA sequences in this region. Single point mutations are sufficient to remove binding capability, providing a rationale for the manner in which different meningococcal PorA escape variants arise. PMID- 16470852 TI - Neural tube development requires the cooperation of p53- and Gadd45a-associated pathways. AB - BACKGROUND: Numerous genetically engineered mouse models for neural tube defects (NTDs) exist, and some of the implicated proteins are functionally related. For example, the growth arrest and DNA damage-inducible protein Gadd45a and tumor suppressor p53 are functionally similar, and both are involved in neural tube development (Gadd45a- and Trp53-null embryos show low levels of exencephaly). To assess their roles in neural tube development, we generated double-null mice from Gadd45a- and Trp53-null mice, as well as from cyclin-dependent kinase inhibitor (Cdkn1a) (p21)-null and xeroderma pigmentosum group C (XPC)-null mice that do not show spontaneous exencephaly. METHODS: Gadd45a-, Trp53-, Cdkn1a-, and XPC-null mice were crossed to generate several double-null mouse models. Embryos (embryonic day [ED] 16-18) from the single- and double-null crosses were scored for NTDs. RESULTS: Deletion of both Gadd45a and Trp53 in mice increased exencephaly frequencies compared to the deletion of either single gene (34.0% in Gadd45a/Trp53-null compared to 8.4% and 9.1% in the Gadd45a- and Trp53-null embryos, respectively). Furthermore, although deletion of another p53-regulated gene, Cdkn1a, is not associated with exencephaly, in conjunction with Gadd45a deletion, the exencephaly frequencies are increased (30.5% in the Gadd45a/Cdkn1a null embryos) and are similar to those in the Gadd45a/Trp53-null embryos. Although XPC deletion increased exencephaly frequencies in Trp53-null embryos, XPC deletion did not increase the exencephaly frequencies in Gadd45a-null embryos. CONCLUSIONS: The increased genetic liability to exencephaly in the Gadd45a/Trp53- and Gadd45a/Cdkn1a-null embryos may be related to the disruption of multiple cellular pathways associated with Gadd45a and p53. PMID- 16470853 TI - A case of methotrexate embryopathy with holoprosencephaly, expanding the phenotype. AB - BACKGROUND: Methotrexate (MTX) embryopathy was described nearly 50 years ago, when this agent began to be used as a cancer treatment and abortifacient. In this report we describe a case with typical features of MTX syndrome together with new features to expand the phenotype. CASE: A 29-year-old woman decided to terminate her unwanted pregnancy because of ill health, as she had conceived soon after her last delivery by cesarian section. At 6 weeks of gestation, she took 2.5 mg of MTX 3 times a day for 7 days. The pregnancy termination failed, and the pregnancy was carried to term. A female infant was delivered who was growth retarded and had characteristic features of MTX embryopathy in addition to holoprosencephaly and other brain malformations, facial hypertrichosis, and long eyelashes- features that have not hitherto been described. CONCLUSIONS: We report the first case of holoprosencephaly in association with MTX exposure during the first 6 weeks of gestation. Physicians and the public should be aware of the effects of MTX on the fetus during pregnancy. PMID- 16470854 TI - Results of longterm hospital based cytological screening in asymptomatic women. AB - Routine cytological screening has been carried out in 27,062 asymptomatic women attending Gynaec and Family Planning O.P.D. of Queen Mary's Hospital, Lucknow, India (April 1971-December 2004). Incidence of squamous intraepithelial lesion (SIL) was found to be 5.9% in the series, while cervical malignancy was seen in 0.6% of cases. The study highlighted the immense utility of cytological screening in minimizing the incidence of carcinoma cervix in the segment of the urban population screened, as the incidence dropped down to 0.5% in the second half from 1.1% noticed in the first half of the screening period. The study also emphasized the utility of clinically downstaging the cervical cancer as 7,316 women showing clinical lesions of cervix were found to harbor SIL in 15.3% and carcinoma cervix in 1.3% of cases as against the incidence of 2.5% for SIL and 0.6% for frank cancer in women with normal cervix. The investigation into different risk factors involved in cervical carcinogenesis revealed that the incidence of SIL and cancer cervix showed a rise with increasing age and parity and prolonged sexual period. The incidences of both cervical cytopathologies were also higher in women of low socio-economic status while religion was found to have no bearing on the occurrence of the disease. Among the four sexually transmitted diseases (STDs) diagnosed in the cervical smears, Trichomonas vaginalis was found to be more prevalent (2.6%), while human papillomavirus (HPV) and Herpes simplex was seen in 0.4 and 0.2% of cases, respectively Herpes simplex was found to have strong affinity with both SIL and carcinoma cervix, while only SIL incidence was high with HPV infection. The study emphasizes need of proper education to women of low socio-economic class for creating awareness regarding hazards and risk factors of cervical cancer as well as management and cure of the disease. PMID- 16470855 TI - Fine-needle aspiration cytology of pulmonary ameloblastic carcinoma of mandibular origin. PMID- 16470856 TI - Intracytoplasmic lumina in invasive micropapillary carcinoma of the lung. AB - Micropapillary carcinoma of the lung is a rare neoplasm, and several reports on micropapillary carcinoma of the lung have been presented to date. We present a case of micropapillary carcinoma of the lung here. A 75-yr-old Japanese man received the medical checkup and his chest X-ray disclosed the abnormal shadow of the lower lobe of the left lung. The histological examination of resected lung and extirpated lymph node showed the finding of micropapillary carcinoma. Some neoplastic cells of primary site contained intracytoplasmic lumina positive for Alcian blue and PAS stains. Pleural effusion appeared 9-mo after the operation. The cytology of pleural effusion showed cohesive clusters of neoplastic cells consisting of 3-20 cells without fibrovascular core. Additionally, intracytoplasmic lumina were observed in some neoplastic cells. Finally, carcinoma cells with micropapillary morphology may possess the intracytoplasmic lumina in the cytoplasm of metastatic site as well as primary site. PMID- 16470857 TI - High-risk human papillomavirus DNA testing: a marker for atypical glandular cells. AB - Cervical/endocervical cytology screening has decreased morbidity and mortality, and implementing adjunctive human papilloma virus (HPV) DNA testing for atypical squamous cells of undetermined significance has improved the specificity for detecting premalignant squamous lesions. Currently, there are no guidelines to perform HPV DNA testing on cervical/endocervical ThinPreps with atypical glandular cells (AGC). To assess the potential role of HPV DNA testing on AGC cases, Hybrid Capture 2 (Digene Corp.) testing was performed on 144 cervical/endocervical AGC specimens. One hundred three of 144 cases had follow up; 60/103 (58.3%) were high-risk HPV negative and 43/103 (42.3%) were high-risk HPV positive. Of 43 HPV-positive patients, 37 had adenocarcinoma in situ (AIS), atypical squamous cells of undetermined significance (ASCUS), or cervical squamous intraepithelial neoplasia, while only one patient without high-risk HPV had a squamous intraepithelial neoplasia. Furthermore, most high-risk HPV positive AGC cases harbored high-grade squamous intraepithelial lesion (HSIL) rather than AIS. Our data support HPV DNA testing of all AGC specimens to detect cervical, especially squamous, neoplasia. PMID- 16470858 TI - Role of fine-needle aspiration cytology in the diagnosis of secondary tumors of the thyroid--twenty years' experience. AB - Secondary tumors of the thyroid, presenting as thyroid nodules although rare, have been documented in literature. This study highlights the frequency, primary sites, and the role of fine-needle aspiration cytology (FNAC) in evaluating secondary tumors of the thyroid at a tertiary care hospital. There were 24 documented cases of secondary tumors of the thyroid from 1982 to 2002 (20 years). The aspirates and histology slides of all these cases were reviewed and the findings noted. On FNAC, 17 cases were diagnosed as secondary tumors, 3 as primary, and in 4 cases it was difficult to categorize the tumors as either primary or secondary. In 12 out of 24 cases, the primary tumor was seen to arise in the larynx (9), and in other sites anatomically close to the thyroid including the trachea (1), and the esophagus (2). Metastases from distant organs were also observed. Secondary tumors of the thyroid are rare and should be suspected in patients when the FNAC shows cytological features not seen in primary thyroid tumors. FNAC alone was required to accurately diagnose secondary tumors of the thyroid in a majority of cases (17/24); however, in some cases ancillary techniques may be required for a definite diagnosis. PMID- 16470859 TI - CDX-2 expression in pulmonary fine-needle aspiration specimens: a useful adjunct for the diagnosis of metastatic colorectal adenocarcinoma. AB - CDX-2 is a homeobox gene, which encodes a transcription factor that plays a vital role in the development and differentiation of intestinal epithelial cells. Recent studies showed that CDX-2 protein expression by immunohistochemistry (IHC) has a high predictive value for confirming the diagnosis of colorectal adenocarcinoma (ACA). The differentiation of primary pulmonary ACA from metastatic colorectal ACA can be extremely challenging on fine-needle aspiration (FNA). The results of immunostains for TTF-1, CK 7, and CK 20 may be controversial in the differential diagnosis. In this study, we evaluated the clinical utility of CDX-2 in lung FNA specimens. Cell blocks from 41 primary lung and 20 colorectal ACAs metastatic to the lungs were retrieved. Cell block sections were immunostained with a CDX-2 monoclonal antibody. Sensitivity and specificity of CDX-2 for colorectal ACA was 75 and 100%, respectively. All five cases negative for CDX-2 were moderately to poorly differentiated colorectal ACAs. Our study confirms that CDX-2 is a highly useful immunohistochemical marker for the differentiation of primary pulmonary ACA from colorectal ACA metastatic to the lungs in FNA specimens. Of diagnostic importance is the loss of CDX-2 immunoreactivity in poorly differentiated colorectal ACAs. PMID- 16470860 TI - Polyoma virus infection is a prominent risk factor for bladder carcinoma in immunocompetent individuals. AB - Despite various reports of BK viral (BKV) DNA sequences or proteins in tumors of the urogenital tract, there has been no study statistically linking infection by this polyoma virus (PV) to tumor development. All PV are potential transforming viruses, the large T-antigen of which interacts with tumor suppressor proteins. Here, we have performed a cross-sectional study of 3,782 patients having had urine cytologic analyses, comparing those diagnosed with PV infection with those not so diagnosed. In order to focus on immunocompetent individuals, renal transplant patients, for whom a diagnosis of PV infection followed immunosuppressive therapy, were excluded. Among the 133 immunocompetent patients diagnosed with PV infection, the most frequently occurring neoplasms were bladder carcinoma (15.8%) and prostate carcinoma (3.8%). The incidence of bladder carcinoma was sufficient to statistically establish temporality in a two-sided test, linking a prior diagnosis of PV infection to a subsequent diagnosis of bladder carcinoma (odds ratio = 3.419, P < 0.001). PMID- 16470861 TI - Fine needle aspiration biopsy of epithelioid hemangioendothelioma of the oral cavity: report of one case and review of literature. AB - Epithelioid hemangioendothelioma (EHE) is an uncommon vascular tumor with biological behavior intermediate between hemangioma and angiosarcoma. It rarely occurs in the oral cavity. We report a case of an 81-yr-old woman with a 2-mo history of a 2 x 2 cm2 submucosal buccal mass. Fine needle aspiration (FNA) smears were paucicellular and showed mainly single atypical large epithelioid cells in a bloody background. The atypical cells had abundant dense cytoplasm, some with fine vacuoles. Occasionally, cells with large cytoplasmic lumina were seen. Cytology preparations from fresh tissue received for frozen section revealed numerous neoplastic cells with large intracytoplasmic lumina, some of which contained red blood cells. In addition, cells with distinct intranuclear inclusions were present. Histologic sections and immunohistochemical stains confirmed the diagnosis of EHE. Although the histologic features of EHE are well recognized, reports of FNA cytology findings are sparsely existent in the literature as several case reports. The characteristic cytological features of EHE are reviewed in this report. We believe that the diagnosis of this rare tumor can be suggested when an adequate FNA specimen is obtained. PMID- 16470862 TI - Specimen adequacy and diagnostic specificity of ultrasound-guided fine needle aspirations of nonpalpable thyroid nodules. AB - Ultrasound-guided fine needle aspiration (USG-FNA) is a safe, effective, and dependable test used to assess thyroid nodules. However, the size of the lesion can adversely affect the outcome of the procedure. The aim of this study was to assess specimen adequacy and diagnostic specificity in USG-FNA of thyroid nodules measuring < or = 1.5 cm. A total of 219 thyroid FNAs were performed in a one year; 26 were obtained by pathologists, 139 by clinicians, and 54 by radiologists under ultrasound guidance. Of the 54 ultrasound-guided aspirates, 19 cases (35%) were performed on nodules < or = 1.5 cm (range 0.8-1.5 cm, mean 1.3 cm). Cytologic material from these 19 cases was reviewed along with corresponding available follow-up surgical material. Standard criteria for specimen adequacy and established morphologic criteria for diagnostic specificity were assessed in each case. All 19 cases met criteria for specimen adequacy, and in 17 cases (89%) specific cytologic diagnoses were made (cellular/adenomatous nodule--2 cases, colloid nodule--10 cases, Hashimoto's thyroiditis--4 cases, and papillary cystic carcinoma--1 case). The diagnoses were confirmed by surgical follow-up in six cases including the case of papillary carcinoma. One case diagnosed as suspicious for a papillary carcinoma subsequently was found to be a follicular adenoma by histology. In one case, a diagnosis of lymphocytic thyroiditis versus intrathyroidal lymphoid tissue was made (See Table I). In majority of cases of USG-FNA of nonpalpable thyroid nodules, adequate material may be obtained for a specific cytopathologic diagnosis. PMID- 16470863 TI - Role of FNA in the medical management of minimally enlarged thyroid. AB - Fine-needle aspiration (FNA) of thyroid is a cost-effective and simple diagnostic tool in the initial screening of patients with thyroid nodules. But, its role in a clinically normal thyroid or a minimally enlarged thyroid in a symptomatic patient suspected of having thyroid dysfunction is unknown. With our 2-yr experience in a setup of a tertiary health care hospital in a developing country, we have aimed to implement a management protocol using FNA thyroid done without ultrasound guidance and TSH estimation done during the same visit in symptomatic patients suspected of having thyroid pathology but presenting with no goiter or having minimally enlarged thyroid with no palpable nodules. The thyroid enlargement in 172 cases were graded with the criteria endorsed by WHO, Pan American Health Organization, and International Council for Control of Iodine Deficiency Disorders into grade 0, 1, and 2. The cases were evaluated cytologically and correlated with TSH values according to the algorithm formulated by the authors. FNA was diagnostic in 86.6, 98, and 100% in grade 0, 1, and 2 goiters, respectively. 52.3% (n = 90), 19.8% (n = 34), 16.9% (n = 29) of cases were diagnosed as Hashimoto's thyroiditis (HT), colloid goiter (CG), and lymphocytic thyroiditis (LT). Sixteen had a combination of LT and CG (n = 6), HT and CG (n = 6), papillary carcinoma (n = 2), and diffuse hyperplasia (n = 2). No statistically significant difference (P = 0.4586) was noted between the groups of patients with grade 0 and grade 1-2 goiter, who underwent FNAC. 38.95% of patients (n = 67) with TSH values greater than 10 microIU/ml and considered hypothyroid showed features of HT/LT at FNA. 23.83% (n = 41) having TSH values between 5 and 10 microIU/ml (subclinical hypothyroidism) also showed features of HT/LT at FNA. Both groups were treated with thyroxine. 35.46% (n = 61) of cases with TSH values within normal range (0.5-5.1 microIU/ml) and considered euthyroid showed a spectrum of lesions at cytology other than HT and LT. They are being followed up to detect them at an early stage of subclinical hypothyroidism. Only 13 cases (7.5%) who were serologically euthyroid showed HT/LT and are being followed-up. Thus, the authors advocate FNA of the thyroid as a single simple cost-effective office procedure in the medical management of all nonpalpable/minimally enlarged thyroid in patients suspected of having thyroid pathology and/or in combination with TSH values. FNA helps in early detection of subclinical hypothyroidism, which is of utmost importance in pregnant women and further makes possible availability of baseline values for future reference. With the implementation of this protocol of FNA thyroid without imaging, we affirm that the practice of cytology which differs in different geographic areas and from country to country depending on economy and availability of infrastructure can be made more user-friendly. PMID- 16470864 TI - Nasopharyngeal biopsy imprint cytology: a retrospective analysis of 191 cases. AB - Patients with nasopharyngeal carcinoma (NPC) are common in Taiwan. To provide efficient management to patients, the surgeons often perform cytological imprints immediately after biopsies of lesions suspicious for NPC. The results of cytological assessment of imprints usually are reported within 30 min after biopsies. The patients with positive cytological results can then be arranged for further examinations during the same visit. We reviewed 191 imprints and corresponding biopsies from 187 patients during 1997-2004 at Koo Foundation Sun Yat-Sen Cancer Center, Taipei. The cytological diagnoses were categorized into four groups: negative (62 cases), suspicious (8 cases), positive (116 cases), and inadequate specimen (5 cases). There were 18 false-negative and 1 false-positive diagnoses. All suspicious cases were positive histologically. Our results showed a sensitivity of 87.2% and a specificity of 97.8%. The accuracy was 89.8%. Therefore, nasopharyngeal imprint cytology is a sensitive and specific method for rapid diagnosis of nasopharyngeal cancer at an outpatient setting. PMID- 16470865 TI - Making the cut: what can be regularly and reliably identified in gynecologic cytology? PMID- 16470866 TI - Protocol for the processing of bloody cervical specimens: glacial acetic acid and the ThinPrep Pap Test. AB - Since the utilization of the ThinPrep Pap Test in clinical practice, laboratories have strived to optimize cellular yields from cervicovaginal samples. Bloody specimens that often hamper cytologic interpretation and/or render a specimen unsatisfactory for diagnosis on conventional smears also affect ThinPrep preparations. Currently, there is no uniformly utilized method for the processing of bloody cervical specimens sent for analysis by the ThinPrep method. The purpose of this study is to present the protocol utilized by the University of Wisconsin Hospital and Clinics. PMID- 16470867 TI - Cytologic diagnosis of brain metastasis from hepatocellular carcinoma by squash preparation. AB - Hepatocellular carcinoma (HCC) metastasizes to the brain is rare instances. In published series and case reports of metastatic HCC, diagnosis of central nervous system metastases has been determined by histologic methods. We present a case of metastatic HCC of brain diagnosed by squash cytologic preparation. A 69-year-old male, HCV positive, suffering from post-hepatitic cirrhosis, initially diagnosed at age 68 with HCC presented with headaches of increasing frequency and severity. A computed tomography scan confirmed a 3-cm nodule in the right parietal lobe of the brain. Squash cytology was performed intraoperatively and preparations of a small tissue fragment resected from the mass showed medium-to-large-sized, well cohesive clusters or sheets of uniform tumor cells. The tumor cells are highly cellular and contain solitary tumor cells in loose groupings as well as many fragments. They also appear somewhat bizarre and contain large, round, or ovoid nuclei with prominent nucleoli. Cytologic diagnosis of metastatic HCC was rendered reported and confirmed by a subsequent frozen section examination. To the best of our knowledge, this is the first reported case in which HCC was reported as brain metastasis, by using squash cytology. We suggest that intraoperative squash cytologic examination be viewed as a useful initial approach in the diagnosis of metastatic brain tumor. PMID- 16470868 TI - Diffuse sclerosing variant papillary carcinoma of the thyroid: report of four cases with fine-needle aspirations. PMID- 16470869 TI - Genetic identification of AChE as a positive modulator of addiction to the psychostimulant D-amphetamine in zebrafish. AB - Addiction is a complex maladaptive behavior involving alterations in several neurotransmitter networks. In mammals, psychostimulants trigger elevated extracellular levels of dopamine, which can be modulated by central cholinergic transmission. Which elements of the cholinergic system might be targeted for drug addiction therapies remains unknown. The rewarding properties of drugs of abuse are central for the development of addictive behavior and are most commonly measured by means of the conditioned place preference (CPP) paradigm. We demonstrate here that adult zebrafish show robust CPP induced by the psychostimulant D-amphetamine. We further show that this behavior is dramatically reduced upon genetic impairment of acetylcholinesterase (AChE) function in ache/+ mutants, without involvement of concomitant defects in exploratory activity, learning, and visual performance. Our observations demonstrate that the cholinergic system modulates drug-induced reward in zebrafish, and identify genetically AChE as a promising target for systemic therapies against addiction to psychostimulants. More generally, they validate the zebrafish model to study the effect of developmental mutations on the molecular neurobiology of addiction in vertebrates. PMID- 16470870 TI - Influence of water activity in the synthesis of galactooligosaccharides produced by a hyperthermophilic beta-glycosidase in an organic medium. AB - The present study evaluated the influence of water activity and lactose concentration on the synthesis of galactooligosaccharides (GOS), by means of a hyperthermophilic beta-glycosidase in an organic system. The production of GOS gradually grew as water activity increased in the reaction system; later, their synthesis decreased as water activity increased. The authors used the response surface methodology to study how different water activities and different concentrations of lactose influenced the synthesis of GOS and their length. In every case, the variable that proved to have the greatest effect on GOS synthesis was water activity. Maximum GOS3 synthesis was reached at a water activity interval of 0.44-0.57, with lactose concentrations of 0.06%-0.1%, while GOS4 and GOS5 maxima were reached at water activity intervals of 0.47-0.57 and 0.49-0.60, respectively. The research showed that higher water activity was required to synthesize GOS of greater length. Synthesis of GOS would then depend on the flexibility of the enzyme, which in turn would depend on water activity of the reaction system. This hypothesis was supported by experiments in which the reaction temperature was modified in order to change the flexibility of the enzyme, thus leading to longer GOS. PMID- 16470871 TI - Potential of the marine sponge Hymeniacidon perleve as a bioremediator of pathogenic bacteria in integrated aquaculture ecosystems. AB - The aim of this article is to investigate the potential of using sponges as a bioremediator to remove pathogenic bacteria in integrated aquaculture ecosystems. Using the inter-tidal marine sponge Hymeniacidon perleve as a model system, the ability of removing the most common pathogens Escherichia coli and Vibrio anguillarum II in aquaculture waters was screened in laboratory tests. In sterilized natural seawater (SNSW) supplemented with E. coli at (7.0-8.3) x 10(6) cells/mL, H. perleve can remove an average 96% of E.coli within 10.5 h at a filter rate of ca. (7.53-8.03) x 10(7) cells/h x g of fresh sponge in two independent tests. Despite the removal efficiency and filter rate are similar; the clearance rates (CR) vary significantly among individual sponge specimens and between two batches. For the tests on V. anguillarum II in SNSW, about 1.5 g fresh sponges can keep the pathogen growth under control at a lower initial density 3.6 x 10(4) cells/mL of 200 mL water volume. Further tests were done for 24 h using about 12 g fresh sponge in 2-L actual seawater collected from two aquaculture sites that have ca. eightfold difference in pathogenic bacteria load. The concentrations of E. coli, Vibrio, and total bacteria at 24 h in treatment groups were markedly lower, at about 0.9%, 6.2%-34.5%, and 13.7%-22.5%, respectively, of those in the control. Using a fluoresce stain 1,1'-dioctadecyl 3,3,3',3'-tetramethylindocarbocyanine perchlorate, E. coli, and V. anguillarum II cells were stained and fed to sponges in two independent tests. The confocal microscope observation confirmed that the sponges filtering-retained and digested these bacteria by phagocytosis. PMID- 16470872 TI - Influence of perfusion on metabolism and matrix production by bovine articular chondrocytes in hydrogel scaffolds. AB - Articular cartilage has a limited capacity for self-repair after damage. Engineered cartilage is a promising treatment to replace or repair damaged tissue. The growth of engineered cartilage is sensitive to the extracellular culture environment. Chondrocytes were seeded into alginate beads and agarose scaffolds at 4 millions/mL, and the response to static and perfusion culture was examined over period of up to 12 days. For both types of scaffolds, the chondrocytes kept their differentiated morphology over 12 days in all culture conditions. In alginate beads, more glycosaminoglycans (GAGs) were produced in perfusion culture than in static conditions. GAG distribution in alginate constructs was more uniform in perfusion culture than in static culture. However, in agarose constructs there was no significant difference in GAG production between static culture and perfusion culture. Under perfusion culture, the retention rate of GAG in alginate was higher than in agarsoe. It is suggested that the positive effect of perfusion culture only can be achieved by an appropriate choice of other factors such as scaffold materials. PMID- 16470873 TI - Affinity-tagged green fluorescent protein (GFP) extraction from a clarified E. coli cell lysate using a two-phase aqueous micellar system. AB - Green fluorescent protein (GFP) has been proposed as an ideal choice for a protein-based biological indicator for use in the validation of decontamination or disinfection treatments. In this article, we present a potentially scalable and cost-effective way to purify recombinant GFP, produced by fermentation in Escherichia coli, by affinity-enhanced extraction in a two-phase aqueous micellar system. Affinity-enhanced partitioning, which improves the specificity and yield of the target protein by specific bioaffinity interactions, has been demonstrated. A novel affinity tag, family 9 carbohydrate-binding module (CBM9) is fused to GFP, and the resulting fusion protein is affinity-extracted in a decyl beta-D-glucopyranoside (C10G1) two-phase aqueous micellar system. In this system, C10G1 acts as phase forming and as affinity surfactant. We will further demonstrate the implementation of this concept to attain partial recovery of affinity-tagged GFP from a clarified E. coli cell lysate, including the simultaneous removal of other contaminating proteins. The cell lysate was partitioned at three levels of dilution (5x, 10x, and 40x). Irrespective of the dilution level, CBM9-GFP was found to partition preferentially to the micelle rich phase, with the same partition coefficient value as that found in the absence of the cell lysate. The host cell proteins from the cell lysate were found to partition preferentially to the micelle-poor phase, where they experience less excluded-volume interactions. The demonstration of proof-of principle of the direct affinity-enhanced extraction of CBM9-GFP from the cell lysate represents an important first step towards developing a cost-effective separation method for GFP, and more generally, for other proteins of interest. PMID- 16470874 TI - Assessment of alaryngeal speech using a sound-producing voice prosthesis in relation to sex and pharyngoesophageal segment tonicity. AB - BACKGROUND: A pneumatic artificial sound source incorporated in a regular tracheoesophageal shunt valve may improve alaryngeal voice quality. METHODS: In 20 laryngectomees categorized for sex and pharyngoesophageal segment tonicity, a prototype sound-producing voice prosthesis (SPVP) is evaluated for a brief period and compared with their regular tracheoesophageal shunt speech. RESULTS: Perceptual voice evaluation by an expert listener and acoustical analysis demonstrate a uniform rise of vocal pitch when using the SPVP. Female laryngectomees with an atonic pharyngoesophageal segment gain vocal strength with the SPVP. Exerted tracheal pressure and airflow rate are equivalent to those required for regular tracheoesophageal shunt valves. However, communicative suitability and speech intelligibility deteriorate by the SPVP for most patients. Tracheal phlegm clogging the SPVP is a hindrance for most patients. CONCLUSIONS: The SPVP raises vocal pitch. Female laryngectomees with an atonic or severely hypotonic pharyngoesophageal segment can benefit from a stronger voice with the SPVP. PMID- 16470876 TI - Supracricoid laryngectomy with cricohyoidoepiglottopexy for advanced glottic cancer. AB - BACKGROUND: Supracricoid laryngectomy with cri-cohyoidoepiglottopexy (CHEP) is a conservative surgical procedure indicated in selected cases of advanced glottic carcinoma. METHODS: This study is a review of our experience with 43 patients with T3/T4 glottic squamous cell carcinoma who underwent CHEP in our institution. All but two patients underwent selective neck dissections. All patients were staged on the basis of the 2002 TNM classification. Rates of recurrence and death were estimated by the Kaplan-Meier method. RESULTS: The 5-year disease-specific survival and 5-year relapse-free survival rates were 78% and 83%, respectively. Neck metastases were found in three patients. Cartilage invasion occurred in 11 cases. The average length of hospital stay was 5.7 days. The mean time of enteral feeding tube was 33.8 days, and the mean time for tracheotomy was 29.6 days. Overall, normal swallowing was achieved in 74.4% of patients. Eleven patients had mild and major complications. Laryngeal stenosis emerged as the most frequent major complication. Three patients (6.9%) had local recurrences. Two patients (4.6%) had neck metastases. CONCLUSIONS: On the basis of this study, over a 7 year period with 43 patients with advanced glottic cancer, a successful on cologic outcome is confirmed. PMID- 16470877 TI - Inflammatory myofibroblastic tumor of the larynx. AB - BACKGROUND: Inflammatory myofibroblastic tumor, composed of myofibroblastic spindle cells with acute and chronic inflammatory cells, is an unusual, benign solid mass that mimics a neoplastic process. METHODS: We report a rare case of a patient with a laryngeal inflammatory myofibroblastic tumor. Laryngoscopy demonstrated a submucosal mass involving the right false cord. The mass was a well-enhanced supraglottic lesion on CT scan. It showed medially high signal intensity and peripherally low signal intensity on T2-weighted MR images, and it displayed a high magnetization transfer ratio; before surgery, it was believed to be a malignant tumor. Laryngoscopic biopsy was performed. Pathologic features of the specimen were diagnostic for inflammatory myofibroblastic tumor. RESULTS: Steroid therapy was chosen for further treatment. No recurrence was observed for 4 years. CONCLUSION: In patients with chronic hoarseness who have a malignant looking submucosal laryngeal mass, inflammatory myofibroblastic tumor should be considered. Conservative surgery and steroid treatment are advocated because of laryngeal preservation. PMID- 16470875 TI - Matrix metalloproteases in head and neck cancer. AB - Matrix metalloproteases (MMPs) are a collection of enzymes capable of cleaving extracellular matrix components, growth factors, and cell-surface receptors. MMPs modulate most aspects of tumorigenesis and are highly expressed in cancer compared with normal tissues. Preclinical studies have demonstrated that head and neck squamous cell carcinomas (HNSCCs) express high levels of MMPs in vivo and that inhibition of these enzymes in vitro and in mouse models decreases invasion and metastasis. However, the clinical trials for MMP inhibitors have failed to demonstrate a significant survival advantage in most cancers. The disparity between preclinical and clinical studies has led to the reevaluation of how MMP functions in cancer and the design of clinical trials for molecularly targeted agents. Mouse model data and analysis of HNSCC tumor specimens suggests that membrane type-1 MMP (MT1-MMP) may be a critical enzyme in tumor cell invasion and survival in vivo. This accumulated data provide evidence for development of selective MT1-MMP inhibitors as therapy in HNSCC. PMID- 16470878 TI - Distinct array comparative genomic hybridization profiles in oral squamous cell carcinoma occurring in young patients. AB - BACKGROUND: Oral cancer typically affects smokers older than 50 years of age. Recently, however, a marked increase in the number of patients 40 years old and younger, many with no history of tobacco smoking, has been noted. Studies in this age group have so far been restricted to genomic areas well recognized as abnormal in typical patients with oral cancer. The aim of this study was to assess genomic aberrations in oral cancer, using comparative genomic hybridization (CGH) microarray technology, and to compare the genomic aberration profile of patients older than 40 years old with those 40 years old and younger. METHODS: Tumor samples from 20 patients with oral cancer (age range, 21-78; 10 smokers and 10 nonsmokers) were laser microdissected, and array CGH was used to identify genomic imbalances in these two cohorts. RESULTS: The older cohort showed high numbers of gains and losses in contrast to very few copy number changes in the younger nonsmoker cohort. In concurrence with the literature, tumors from the older cohort manifested deletions involving 3p and 9p21 and gains involving 3q, 5q, 7p, 8q, 11q, and 20q. The younger group, particularly the nonsmokers, showed very few changes overall, and the aberrations were not in the sites classically associated with oral cancer. Deletion of CDKN2A (p16) was completely absent in the younger group but was present in 50% of the older cohort. CONCLUSIONS: We have demonstrated that there is far less genomic instability in young nonsmokers with oral cancer than found in typical patients with oral cancer. These observations indicate that oral cancer presenting at a younger age, particularly in nonsmokers, has a genomic profile different from the classically described oral cancer. PMID- 16470879 TI - Clinical utility of somatostatin receptor scintigraphic imaging (octreoscan) in esthesioneuroblastoma: a case study and survey of somatostatin receptor subtype expression. AB - BACKGROUND: For tumors that express somatostatin receptors (SSTR), radiolabeled somatostatin analogs, such as 111In-pentetreotide, can demonstrate the presence of tumor by radioligand uptake using somatostatin receptor scintigraphy (SRS). The use of 111In-pentetreotide for SRS depends on the specific high affinity of octreotide for SSTR subtypes 2, 3, and 5. Of these, SSTR2 has the greatest affinity for octreotide and the greatest relevance for tumor detection with Octreoscan imaging. Discriminating between postoperative changes and residual or recurrent tumor after extensive skull base surgery is often difficult, but in a case of recurrent esthesioneuroblastoma (ENB) we found the use of Octreoscan imaging clinically useful. To better define the general relevance of this imaging technique in this setting, we analyzed SSTR subtype expression in a panel of ENB tumors. METHODS: The case history and correlations between MRI and 111In pentetreotide SRS of a patient with recurrent ENB were reviewed. The expression pattern of the SSTR subtypes in a panel of ENB tumors was then analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR) to better define the potential of more general use of Octreoscan for imaging ENB. To correlate SSTR2 protein expression with 111In-pentetreotide uptake, immunohistochemistry to detect SSTR2 was performed on tumor samples from regions of increased uptake on Octreoscan. RESULTS: The SSTR2 message was expressed at high levels in all five ENB tumor samples, and either SSTR2 protein or histologic findings typical for ENB were found in all tumor tissue obtained from regions of increased 111In pentetreotide uptake. Furthermore, Octreoscan imaging in this case proved useful in clinical decision making. CONCLUSION: The expression pattern of SSTR2 and the specificity of the Octreoscan for regions of active tumor growth support further investigation of the utility of Octreoscan imaging in the diagnosis and surveillance of ENB. Recent advances in novel therapies based on SSTR ligand binding also provide the rationale to consider such novel therapeutic approaches in patients with ENB. PMID- 16470880 TI - Tolerance and adaptation of ethanologenic yeasts to lignocellulosic inhibitory compounds. AB - Synthetic mixtures of predominant lignocellulosic hexose sugars were supplemented with separate aliquots of three inhibitory compounds (furfural, hydroxymethylfurfural (HMF), and acetic acid) in a series of concentrations and fermented by the spent sulfite liquor (SSL)-adapted yeast strain Tembec T1 and the natural isolate Saccharomyces cerevisiae (S. cerevisiae) Y-1528 to compare tolerance and assess fermentative efficacy. The performance of Y-1528 exceeded that of Tembec T1 by a significant margin, with faster hexose sugar consumption, higher ethanol productivity, and in the case of furfural and HMF, faster inhibitor consumption. Nevertheless, furfural had a dose-proportionate effect on sugar consumption rate and ethanol productivity in both strains, but did not substantially affect ethanol yield. HMF had a similar effect on sugar consumption rate and ethanol productivity, and also lowered ethanol yield. Surprisingly, acetic acid had the least impact on sugar consumption rate and ethanol productivity, and stimulated ethanol yield at moderate concentrations. Sequential iterations of softwood (SW) and hardwood (HW) SSL were subsequently inoculated with the two yeast strains in order to compare adaptation to, and performance in lignocellulosic substrates in a cell recycle batch fermentation (CRBF) regime. Both strains were severely affected by the HW SSL, which was attributed to specific syringyl lignin-derived degradation products and synergistic interactions between inhibitors. Though ethanologenic capacity was preserved, a net loss of performance was evident from both strains, indicating the absence of adaptation to the substrates, regardless of the sequence in which the SSL types were employed. PMID- 16470882 TI - Hydromechanical stress in shake flasks: correlation for the maximum local energy dissipation rate. AB - Shake flasks are widely used in biotechnological process research. Bioprocesses for which hydromechanical stress may become the rate controlling parameter include those where oils are applied as carbon sources, biotransformation of compounds with low solubility in the aqueous phase, or processes employing animal, plant, or filamentous microorganisms. In this study, the maximum local energy dissipation rate as the measure for hydromechanical stress is characterized in shake flasks by measuring the maximum stable drop size. The theoretical basis for the method is that the maximum stable drop diameter in a coalescence inhibited liquid/liquid dispersion is only a function of the maximum local energy dissipation rate and not of the dispersing apparatus. The maximum local energy dissipation rate is obtained by comparing the drop diameters in shake flasks to those in a stirred tank reactor. At the same volumetric power consumption, the maximum energy dissipation rate in shake flasks is about 10 times lower than in stirred tank reactors explaining the common observation of considerable differences in the morphology of hydromechanically sensitive cells between these two reactor types. At the same volumetric power consumption, the maximum local energy dissipation rate in baffled and in unbaffled shake flasks is very similar. A correlation is presented to quantify the maximum local energy dissipation rate in shake flasks as a function of the operating conditions. Non negligible drop viscosity may be considered by known literature correlations. Further, from dispersion experiments a critical Reynolds number of about 60,000 is proposed for turbulent flow in unbaffled shake flasks. PMID- 16470881 TI - Chondrogenic differentiation of bovine bone marrow mesenchymal stem cells (MSCs) in different hydrogels: influence of collagen type II extracellular matrix on MSC chondrogenesis. AB - Bone marrow mesenchymal stem cells (MSCs) are candidate cells for cartilage tissue engineering. This is due to their ability to undergo chondrogenic differentiation after extensive expansion in vitro and stimulation with various biomaterials in three-dimensional (3-D) systems. Collagen type II is one of the major components of the hyaline cartilage and plays a key role in maintaining chondrocyte function. This study aimed at analyzing the MSC chondrogenic response during culture in different types of extracellular matrix (ECM) with a focus on the influence of collagen type II on MSC chondrogenesis. Bovine MSCs were cultured in monolayer as well as in alginate and collagen type I and II hydrogels, in both serum free medium and medium supplemented with transforming growth factor (TGF) beta1. Chondrogenic differentiation was detected after 3 days of culture in 3-D hydrogels, by examining the presence of glycosaminoglycan and newly synthesized collagen type II in the ECM. Differentiation was most prominent in cells cultured in collagen type II hydrogel, and it increased in a time dependent manner. The expression levels of the of chondrocyte specific genes: sox9, collagen type II, aggrecan, and COMP were measured by quantitative "Real Time" RT-PCR, and genes distribution in the hydrogel beads were localized by in situ hybridization. All genes were upregulated by the presence of collagen, particularly type II, in the ECM. Additionally, the chondrogenic influence of TGF beta1 on MSCs cultured in collagen-incorporated ECM was analyzed. TGF beta1 and dexamethasone treatment in the presence of collagen type II provided more favorable conditions for expression of the chondrogenic phenotype. In this study, we demonstrated that collagen type II alone has the potential to induce and maintain MSC chondrogenesis, and prior interaction with TGF beta1 to enhance the differentiation. PMID- 16470883 TI - Sequential formation of yellow, red, and orange 1-phenyl-3,3-biphenylene-allene dimers prior to blue tetracene formation: Helicity reversal in trans-3,4-diphenyl 1,2-bis(fluorenylidene)cyclobutane. AB - 1-Phenyl-3,3-biphenyleneallene (2), the base-catalyzed rearrangement product of 9 phenylethynylfluorene (1) yields a yellow, head-to-tail dimer 6 that, upon gentle warming, is converted to the red tail-to-tail isomer trans-3,4-diphenyl-1,2 bis(fluorenylidene)cyclobutane (7), in which the two fluorenylidene moieties severely overlap. The helical sense of the fluorenylidene moieties in 7 matches that of the phenyl substituents, and the interplanar angle between the fluorenylidene moieties is 41 degrees . At 80 degrees C, 6 isomerizes to orange cis-3,4-diphenyl-1,2-bis(fluorenylidene)cyclobutane (8), which at 110 degrees C is converted to orange trans diastereomer 9, whereby the helicity of the overlapping fluorenylidene moieties is reversed from that in 7 such that they are aligned with the ring hydrogen atoms, and the interplanar angle between the fluorenylidene moieties is now 60 degrees . At 180 degrees C, 6 rearranges to dispirodihydrotetracene 3 and blue, electroluminescent diindenotetracene 4, which is readily oxidized to peroxide 5. In the solid state, both 3 and 4 adopt structures with Ci symmetry (only an inversion center) such that the central polycyclic framework is nonplanar. Deprotonation of yellow head-to-tail allene dimer 6 with tBuOK in DMSO and reprotonation with HOAc yields the [1,3]-hydrogen migration product 10, in which the proton originally on the cyclobutane ring is now sited at C9 on the exocyclic fluorenyl substituent. Analogously, deprotonation and reprotonation of orange dimer 9 furnishes [1,3]-hydrogen migration product 11. Side product 17, formed during the synthesis of 1 from 9 phenylethynylfluoren-9-ol, BF3 and Et3SiH, was shown to be a silyl-indene spiro linked to C9 of fluorene. All products were characterized by NMR spectroscopy and X-ray crystallography, and the mechanisms of these interconversions are discussed. PMID- 16470884 TI - Peptide recognition: encapsulation and alpha-helical folding of a nine-residue peptide within a hydrophobic dimeric capsule of a bowl-shaped host. AB - A dimeric capsule of coordination bowl 1 encapsulated a nine-residue peptide (Trp Ala-Glu-Ala-Ala-Ala-Glu-Ala-Trp; 2) within the large hydrophobic cavity in water, and stabilized the alpha-helical conformation of bound 2. An NMR titration experiment revealed that monomeric bowl 1 recognized two Trp residues at the both terminals of 2 through 1/2 = 1:1 to 2:1 complexation. The 1:1 and 2:1 species exist in equilibrium even in the presence of excess 1. It was found that the formation of the 2:1 complex, in which two bowls of 1 wrapped the whole of 2, became dominant by the addition of NaNO3 due to the fact that the enhanced ion strength increased the hydrophobic interaction between Trp residues and the cavity of 1. The alpha-helical conformation of 2 within the dimeric capsule of 1 was elucidated from detailed NOESY analysis. PMID- 16470885 TI - From diphosphane to diphosphodiide gold(III) derivatives of 1,2 diphosphinobenzene. AB - Treatment of 1,2-diphosphinobenzene with [Au(C6F5)3(tht)] leads to the diphosphane derivative [{Au(C6F5)3}(1,2-PH2C6H4PH2)] (1), which further reacts with other pentafluorophenylgold(III) reagents in the presence of acetylacetonate as deprotonating agent to afford phosphane-phosphide complexes. The noncyclic PPN[{Au(C6F5)3}2(1,2-PHC6H4PH2)] (2; PPN = bis(triphenylphosphine)iminium) has been shown to be a useful starting material for the synthesis of higher nuclearity cyclic or noncyclic diphosphide or even diphosphodiide derivatives through similar reactions. The crystal structures of the trinuclear anionic NBu4[{Au(C6F5)3}(1,2-PHC6H4PH){Au(C6F5)2Cl}{mu-Au(C6F5)2}] (3) and the hexanuclear [{Au(C6F5)3}(1,2-PC6H4P){Au(C6F5)3}{mu-M(dppe)M}2] (M = Au (12), Ag (13)) have been established by X-ray diffraction methods, the last complexes having a bicyclic ring containing three intramolecular interactions between the M(I) centres. PMID- 16470886 TI - A photoactivated trans-diammine platinum complex as cytotoxic as cisplatin. AB - The synthesis and X-ray structure (as the tetrahydrate) of the platinum(IV) complex trans,trans,trans-[Pt(N(3))(2)(OH)(2)(NH(3))(2)] 3 are described and its photochemistry and photobiology are compared with those of the cis isomer cis,trans,cis-[Pt(N(3))(2)(OH)(2)(NH(3))(2)] 4. Complexes 4 and 3 are potential precursors of the anticancer drug cisplatin and its inactive trans isomer transplatin, respectively. The trans complex 3 is octahedral, contains almost linear azide ligands, and adopts a layer structure with extensive intermolecular hydrogen bonding. The intense azide-to-platinum(IV) charge-transfer band of complex 3 (285 nm; epsilon=19 500 M(-1) cm(-1)) is more intense and bathochromically shifted relative to that of the cis isomer 4. In contrast to transplatin, complex 3 rapidly formed a platinum(II) bis(5'-guanosine monophosphate) (5'-GMP) adduct when irradiated with UVA light, and did not react in the dark. Complexes 3 and 4 were non-toxic to human skin cells (keratinocytes) in the dark, but were as cytotoxic as cisplatin on irradiation for a short time (50 min). Damage to the DNA of these cells was detected by using the "comet" assay. Both trans- and cis-diammine platinum(IV) diazide complexes therefore have potential as photochemotherapeutic agents. PMID- 16470887 TI - Clean diesel power via microwave susceptible oxidation catalysts. AB - The problem of soot emissions from diesel engines is introduced and the possible solution of combining doped perovskites and microwave (mw) irradiation to "clean up" diesel soot filters is outlined. Eighteen doped perovskite catalysts are synthesized and tested for propane and CO oxidation, which are taken as model components for soot. The activity, selectivity, and SO2 tolerance are compared under conventional heating and mw irradiation. By combining mw irradiation and doped perovskites, one can create "hot spots" on the catalyst, resulting in efficient and selective heating of the active site, as well as less poisoning. Sr doped and Ce-doped manganese perovskites show the highest activity. These catalysts are also the most selective, and have a high mw susceptibility. Optimal SO2 tolerance is displayed by Cr perovskites, from which the La0.8Ca0.2CrO3 combination uniquely converts propane before CO, and therefore can be used to remove >C2 hydrocarbons from a mix with CO. Possible mechanistic scenarios are presented and discussed. PMID- 16470888 TI - Photocatalytic reforming of methanol by spatially separated Pd particles on special TiO2 layers. PMID- 16470889 TI - Fluorescent hydroxyflavone-zeolite nanoparticles: ship-in-a-bottle synthesis and photophysical properties. AB - 3-Hydroxyflavone (3-OHF) was incorporated in zeolite micropores by ship-in-a bottle synthesis. This strategy consists of constructing the molecule by reaction of small precursors within the cavity. 3-OHF molecules exhibit excited-state intramolecular proton transfer (ESIPT) and a tautomeric equilibrium between the 3 OHF-excited structures N* and T*exists. This equilibrium is strongly affected by the protic nature and polarity of the surrounding medium. The textural and spectroscopic characterization of the dye-loaded zeolite colloids enabled the study of the correlation between the optical properties of the dye and the zeolite micropore environment. PMID- 16470890 TI - A generic approach to monofunctionalized protein-like gold nanoparticles based on immobilized metal ion affinity chromatography. PMID- 16470892 TI - Rational design of DNA nanoarchitectures. AB - DNA has many physical and chemical properties that make it a powerful material for molecular constructions at the nanometer length scale. In particular, its ability to form duplexes and other secondary structures through predictable nucleotide-sequence-directed hybridization allows for the design of programmable structural motifs which can self-assemble to form large supramolecular arrays, scaffolds, and even mechanical and logical nanodevices. Despite the large variety of structural motifs used as building blocks in the programmed assembly of supramolecular DNA nanoarchitectures, the various modules share underlying principles in terms of the design of their hierarchical configuration and the implemented nucleotide sequences. This Review is intended to provide an overview of this fascinating and rapidly growing field of research from the structural design point of view. PMID- 16470891 TI - Psychometric properties of a tool for measuring hormone-related symptoms in breast cancer survivors. AB - Hormone-related symptoms are common in breast cancer survivors and many aspects of these symptoms are currently under study. Reliable and valid assessment tools are needed to successfully study hormone-related symptoms in breast cancer survivors; however, no gold standard currently exists for measuring these symptoms. This study evaluated the psychometric properties of a shortened version of the Breast Cancer Prevention Trial (BCPT) symptom checklist in a sample of 803 breast cancer survivors. Principal factor analysis with Promax oblique rotation revealed a five-factor structure, identifying five separate hormone-related symptoms scales: vasomotor symptoms, urinary incontinence, cognitive/mood changes, vaginal symptoms, and weight gain/appearance concern. Hormone-related symptom scale scores differed by demographic and clinical characteristics according to expectations, suggesting that these five scales from the shortened BCPT checklist are reasonably reliable and valid. Symptom scale scores were only weakly correlated with health-related quality of life scores; however, the pattern of results generally supported the validity of the symptom scales. This study adds to the evidence that breast cancer survivors experience a significant number of hormone-related symptoms. Future clinical trials and quality of life and symptom management intervention studies would benefit from accurate assessment of hormone-related symptoms with the five scales from the shortened BCPT checklist. PMID- 16470893 TI - Construction of molecular logic gates with a DNA-cleaving deoxyribozyme. PMID- 16470895 TI - Rational design and first principles studies toward the discovery of a small and versatile proton sponge. PMID- 16470894 TI - Spontaneous colorimetric sensing of the positional isomers of dihydroxynaphthalene in a 1D organogel matrix. PMID- 16470896 TI - UCST wetting transitions of polyzwitterionic brushes driven by self-association. PMID- 16470897 TI - A striking periodicity of the cis/trans isomerization of proline imide bonds in cyclic disulfide-bridged peptides. PMID- 16470898 TI - Sequential catalysis: a metathesis/dihydroxylation sequence. PMID- 16470900 TI - Differences in and comparison of the catalytic properties of heme and non-heme enzymes with a central oxo-iron group. PMID- 16470899 TI - Synthesis of the C1-C26 northern portion of azaspiracid-1: kinetic versus thermodynamic control of the formation of the bis-spiroketal. PMID- 16470901 TI - Temperature-sensitive core-shell microgel particles with dense shell. PMID- 16470902 TI - Syntheses, structures, and reactivity of radial oligocyclopentadienyl metal complexes: penta(ferrocenyl)cyclopentadienyl and congeners. PMID- 16470903 TI - Donor-acceptor metallocene catalysts for the production of UHMW-PE: pushing the selectivity for chain growth to its limits. PMID- 16470904 TI - Towards polymer-based hydrogen storage materials: engineering ultramicroporous cavities within polymers of intrinsic microporosity. PMID- 16470905 TI - 3'-(2,4-Dinitrobenzenesulfonyl)-2',7'-dimethylfluorescein as a fluorescent probe for selenols. PMID- 16470907 TI - Role of the ephrin and Eph receptor tyrosine kinase families in angiogenesis and development of the cardiovascular system. AB - Angiogenesis is a highly complex orchestrated process that plays a critical role in normal development and in the pathophysiology of multiple disease processes, including tumour neovascularization, ischaemic recovery, and wound healing. In recent years there has been a resurgence of interest in Eph receptors and their ligands, ephrins, as their participation in vasculogenesis and angiogenesis has become apparent. The Eph receptor family is the largest family of receptor tyrosine kinases identified to date. The Eph receptors and their membrane anchored ligands, ephrins, are unique in that they mediate bi-directional signalling. This is concomitant with activation of the Eph receptor tyrosine kinase domain and transduction of the typical forward signal into the receptor bearing cell. The ligand-receptor interaction also leads to transduction of a reverse signal into the ephrin-bearing cell. The Eph/ephrin signalling mechanism is responsible for diverse and complex biological functions mediated by Eph receptors and ephrin ligands. These include vascular development, tissue-border formation, cell migration, axon guidance, and synaptic plasticity. The role of Eph receptors and ephrins in the processes of development of the cardiovascular system, angiogenesis, and vascular remodelling has been the subject of intense investigation since they were first identified in 1987. This review addresses the role of this new growth factor receptor tyrosine kinase family in those processes and provides new insights into the way in which Eph receptors and ephrin ligands modulate the angiogenic response and participate in vascular remodelling and vascular boundary formation during development of the cardiovascular system and vascularization of cancer. PMID- 16470908 TI - Re: Ward et al. Routine testing for mismatch repair deficiency in sporadic colorectal cancer is justified. J Pathol 2005;207:377-384. PMID- 16470910 TI - Surgical treatment preferences in Chinese-American women with early-stage breast cancer. AB - Breast-conserving treatment (BCT) is considered preferable to radical or modified radical mastectomy (MRM) for most women diagnosed with early-stage breast cancer. Studies have found that Chinese-American women are more likely to be treated with MRM, regardless of age or stage of disease. The central question posed in this study is: what are the cultural factors that influence the selection of treatments by Chinese-American patients and the presentation of treatment options by providers? Focused group discussions, semi-structured interviews, and ethnographic observations were conducted with 69 Chinese-American women and 14 health professionals. Results indicated that (1) many Chinese-American women, regardless of age, migration status, education, and income level, expressed a preference for MRM because it was seen as safer; (2) this was primarily a communal belief that even women who had BCT felt pressure to conform to; (3) some women felt pressured into the BCT decision by their physician and/or were unhappy with their choice; (4) patient and physician bias and difficulties with translation issues and supplemental materials all led to cross-cultural miscommunication; and (5) this miscommunication was cited as a reason for (a) not completing treatment regimes or (b) drastic changes in the treatment selected. PMID- 16470912 TI - [Effects of phytochemicals on cytochrome P450]. PMID- 16470914 TI - [Kaempferol activates human steroid and xenobiotic receptor-mediated cytochrome P450 3A4 transcription]. AB - OBJECTIVE: To investigate whether kaempferol stimulates pregnane X receptor (PXR) mediated transcription of CYP3A4. METHODS: Transient cotransfection reporter gene assay was performed with PXR expression plasmid and a reporter plasmid containing the XREs in the CYP3A4 gene promoter in HepG(2)cells. RESULTS: Kaempferol activated PXR-mediated transcription of CYP3A4 in a dose, time-dependent manner. In the dose-response study, kaempferol exposure at concentrations of 1.0 x 10( 3), 1.0 x 10(-2), 0.1, 1.0 and 10.0 mol/L for 24 h increased CYP3A4 transcription by (1.31+/-0.27), (1.45+/-0.36), (1.96+/-0.50), (2.90+/-1.07) and (7.93+/-0.75) fold, respectively compared with 0.1% DMSO (P<0.05). The results from time-course study showed that after 48 h exposure 1.0 and 10.0 mol/L of kaempferol enhanced the transcription of CYP3A4 by (3.73+/-1.21) fold and (8.42+/-1.47) fold, respectively. CONCLUSION: Kaempferol may be a human CYP3A4 gene inducer through PXR, and may affect the metabolism of a large number of substrates of CYP3A4 simultaneously taken. PMID- 16470913 TI - [Human pregnane X receptor-mediated transcriptional regulation of cytochrome P450 3A4 by some phytochemicals]. AB - OBJECTIVE: To test the effect on human pregnane X receptor (hPXR)-mediated transcription regulation of CYP3A4 by five selected phytochemicals. METHODS: Transient cotransfection reporter gene assays in HepG(2) cells were performed with the hPXR expression plasmid and the reporter gene plasmid which contains XRE in the promoter of CYP3A4 linked to luciferase. RESULTS: In the dose-effect study, soybean isoflavone, luteolin and curcumin induced the CYP3A4 transcription via PXR in an evident dose-dependent manner, but isorhamnetin and rutin did not. The inducibility of soybean isoflavone, luteolin and curcumin was also increased in concentrations between 1 micromol/L and 50 micromol/L, 24 h after induction, 50 micromol/L soybean isoflavone, luteolin and curcumin exhibited a 5.46-fold, 2.87-fold, and 2.07-fold increase respectively, compared with 0.1% DMSO treated cells. In the time-effect study, 10 micromol/L and 50 micromol/L soybean isoflavone, luteolin and curcumin induced CYP3A4 transcription between 12 h and 48 h, the strongest induction appeared in 48 h. 48 h after induction, 50 micromol/L soybean isoflavone, luteolin and curcumin exhibited a 6.72-fold, 3.24 fold, and 2.13-fold increase respectively, compared with 0.1% DMSO treated cells. CONCLUSION: Three phytochemicals, i.e. soybean isoflavone, luteolin and curcumin stimulate the PXR-mediated transcription of CYP3A4. Isorhamnetin and rutin have no effect on the CYP3A4 transcription via PXR. PMID- 16470915 TI - [Effects of kaempferol and quercetin on cytochrome 450 activities in primarily cultured rat hepatocytes]. AB - OBJECTIVE: To observe the effects of kaempferol and quercetin on the activity of cytochrome P450 in rat hepatocytes. METHODS: Primarily cultured rat hepatocytes were exposed to kaempferol or quercetin in concentrations of 0.1, 1, 10 micromol/L for 12 h, 24 h and 48 h. Hepatocytes CYP isoemzymes-erythromycin N demethylase (ERND) and aminopyrine N-demethylase (ADM) activities were determined by Nash methods. Erythromycin (10 micromol/L) was used as positive control and DMSO(0.1%) as solvent control. RESULTS: Kaempferol and quercetin inhibited ENRD activity in a dose-and time-dependent manner. In dose-response study, the ENRD activities in kaempferol (0.1,1 and 10 micromol/L) treated groups were (0.088+/ 0.008), (0.074+/-0.006) and (0.041+/-0.003)micromol/(mg.min(-1)), respectively. ENRD activity in quercetin treated groups at the same concentrations were (0.082+/-0.007), (0.063+/-0.007) and (0.034+/-0.005) micromol/(mg.min(-1)), respectively. In time-courses study, the ENRD activity exposed to 10 micromol/L kaempferol or quercetin for 12 h and 48 h were (0.053+/-0.006) and (0.037+/ 0.007) micromol/(mg.min(-1)), or (0.067+/-0.005) and (0.032+/-0.004) micromol/(mg.min(-1)). ADM activity was inhibited only by kaempferol in 10 mol/L at 24 h, but was not significantly altered by quercetin at any concentration tested. CONCLUSION: In the present condition, kaempferol and quercetin act as potential CYP3A4 inhibitors as they can significantly inhibit ENRD in primarily cultured rat hepatocytes. PMID- 16470916 TI - [Effect of kaempferol on the pharmacokinetics of nifedipine in rats]. AB - OBJECTIVE: To investigate the effect of kaempferol on the pharmacokinetics of nifedipine (NFP) in rats. METHODS: Twenty male SD rats, weighing 220-260 g, were distributed randomly into 4 groups. The animals were fasted, but allowed free access to water for 12 h before the administration of drugs. NFP dissolved in corn oil was administered via gastric intubation to the rats in control group at a dose of 10 mg/kg. Kaempferol was administered orally to the other three groups with dose of 5, 10, 15 mg/kg, respectively, followed by oral administration of NFP 10 mg/kg. Blood samples were collected through tail vein in heparinized plastic microcentrifuge tubes before and after drug administration. The plasma concentration of NFP was monitored with reversed phase high-performance liquid chromatography (RP-HPLC). Nimodipine was used as the internal standard. Statistical data evaluation was performed with Student's t-test and one-way analysis of variances. RESULTS: The maximal plasma concentration (C(max)) of the three treated groups were 0.51, 0.70 and 0.81 microg/ml, respectively. The area under the concentration-time curve (AUC(0-8)) were 1.81, 2.83 and 3.63 microg/(h.ml(-1)), respectively. The C(max), AUC(0-8) and the mean retention time (MRT(0-8)) of NFP were significantly increased by simultaneous oral treatment with kaempferol (P<0.01). On the other hand, there were no significant differences in the mean peak value time in plasma (T(max)) and the elimination half-life (t1/2(ke)) between the control and the treated groups. CONCLUSION: The concomitant oral use of kaempferol with NFP may influence the pharmacokinetic parameters of NFP in rats, which suggests that kaempferol might reduce the first pass metabolism of NFP. PMID- 16470917 TI - [Comparison of vasodilatation effect between quercetin and rutin in the isolated rat thoracic aorta]. AB - OBJECTIVE: To determine the possible difference in vasodialtation effect of quercetin and rutin. METHODS: The isolated rat thoracic aorta was treated with phenylephrine (PE), and the effects of quercetin and rutin on the preconstricted aorta rings with or without endothelium were determined by organ bath technique. Nitric oxide synthase inhibitor L-N(G)-nitroarginine methyl-ester (L-NAME), guanylyl cyclase inhibitor methylene blue, cyclooxygenase inhibitor indomethacin were used to explore the mechanism. RESULTS: Quercetin (10-160 micromol/L) caused vasorelaxation of aorta rings preconstricted with PE in endothelium-intact and denuded aorta rings in a dose-dependent manner. Rutin(10-160 micromol/L) caused dose-dependent vasorelaxation in endothelium-intact rings preconstricted with phenylephrine, but not in denuded aorta rings. The maximal response (Rmax) values calculated from vasorelaxation curves of quercetin and rutin were (77.20+/-6.11)% and (44.28+/-7.48)%, respectively. There was no difference between median effective concentration (EC(50)) values of quercetin and rutin. Pretreatment with L-NAME (0.1 mmol/L) abolished the vasorelaxation by rutin,but did not influence the vasodilating effect of quercetin in endothelium-intact rings. Pretreatment with methylene blue (10 mmol/L) canceled the vasorelaxation both by quercetin and rutin. Pretreatment with indomethacin (10 micromol/L) attenuated the vasodilatation of quercetin, but did not affect the vascular effect of rutin. CONCLUSION: The vasodilatation effect of quercetin is more potent than rutin. The vasodilatation effect of quercetin might be mediated by guanylyl cyclase and cyclooxygenase-dependent pathway, while the vasodilatation by rutin might be via nitric oxide-guanylyl cyclase pathway. PMID- 16470918 TI - [Protection of Hsian-tsao (Mesona procumbens Hemsl)on H2O2-induced DNA damage of primarily cultured mice spleen lymphocytes]. AB - OBJECTIVE: To investigate the protection of Hsian-tsao(Mesona procumbens Hemsl)on H(2)O(2)-induced DNA damage of primarily cultured spleen lymphocytes. METHODS: Spleen lymphocytes were primarily cultured for 24 h and then divided into 6 groups randomly: 4 groups were treated with different concentrations of Hsian tsao extracts for 60 min, then with 50 micromol/L H(2)O(2); 1 group was treated with 50 micromol/L H(2)O(2) only and the last group with PBS. Twenty min after H(2)O(2)or PBS treatment SCGE and laser confocal microscopy were performed, then the comet length and percentage of cells with migrated DNA were measured. RESULTS: H(2)O(2) caused severe DNA damage in primarily cultured spleen lymphocytes and Hsian-tsao extracts reduced the DNA damage at a concentration of 10, 50 and 100 microg/ml. The quantity of comet cells was reduced from 100% to 88%, 60%, 36% (P<0.05, 0.01, 0.01) and the comet length was reduced from (49.56+/ 6.94) microm to (41.14+/-5.64) microm, (38.89+/-9.81) microm, (28.62+/-4.66) microm (P<0.05, 0.01, 0.01), respectively. CONCLUSION: Hsian-tsao has dramatic ability of antioxidant. It can protect cells from the damage of ROS-induced DNA injury in the certain range. PMID- 16470919 TI - [Detection, enrichment and expansion of T lymphocytes mediating alloresponse based on cytokine]. AB - OBJECTIVE: To detect, enrich and expand the cytokine secreting T lymphocytes after allogeneic PBMNCs stimulation. METHODS: The novel cytokine secretion assay (CKSA) was applied to detect T lymphocytes secreting IFN-gamma at single cell level in human mixed lymphocytes reaction. IFN-gamma secreting T cells were enriched by means of magnetic sorting system and expanded with OKT(3), anti CD(3)mAb and IL-2 combination. Antigen specificity of the expanded cells was confirmed using enzyme linked immunospot assay. RESULTS: A sizable proportion of IFN-gamma secreting T lymphocytes could be detected [(1.12 +/-0.13)% compared with (0.23 +/-0.07)%] and be further enriched to (67.3 +/-10.5)%, or (93.8 +/ 22.1) fold. T lymphocytes could be expanded up to 600-fold within 21-28 days and the specific IFN-gamma response of expanded cells was confirmed with stimulation of the relevant allogeneic PBMNC, which was significantly higher than the irrelevant PBMNC control. CONCLUSION: It is feasible to detect significantly increased IFN-gamma secreting T lymphocytes after allogeneic PBMNCs stimulation based on the CKSA technique at single cell level and these cells can be efficiently enriched and expanded for further research. PMID- 16470920 TI - [Activity of the TNF-related apoptosis-inducing ligand gene expressed from the hTERT promoter on colon cancer cell line HT-29]. AB - OBJECTIVE: To evaluate the expression and activity of TNF-related apoptosis inducing ligand (TRAIL) gene expressed from the hTERT promoter on colon cancer cell line HT-29. METHODS: GFP/TRAIL gene expressed from the hTERT promoter was transfected into HT-29 with adenoviral vectors system, expression and apoptosis inducing ability of GFP/TRAIL protein were determined with fluorescence-activated cell sorting (FACS) method. RESULTS: The expression of GFP gene was 31.4 % and 67.0 % with either hTERT promoter or CMV promoter in DLD1 cells; GFP/TRAIL gene was able to inhibit cell growth (74.2%) and induce apoptosis (25.8%) of HT-29 cells. There was significant difference between Ad/hTERT-gTRAIL and the other two control groups (PBS and Ad/CMV-GFP, P<0.05). CONCLUSION: The GFP/TRAIL gene with hTERT promoter transfected by adenoviral vector was successfully expressed in HT 29 cell, which can both inhibit cell growth and induce apoptosis of colon cancer cell line HT-29. PMID- 16470921 TI - [Apoptosis induces by exogenous nitric oxide in Tca8113 cells]. AB - OBJECTIVE: To observe the apoptosis induced by exogenous NO in Tca8113 cells and to investigate the possible mechanism. METHODS: SNP as NO donor was used to treat the tongue squamous cell carcinoma Tca8113 cells. Cytotoxic and apoptotic effects of NO on Tca8113 cells were examined by using MTT assay, acridine orange (AO) staining, Wright-Giemsa staining, agarose gel electrophoresis and flow cytometry. Western blot was performed for investigating the apoptotic mechanism. RESULTS: NO had a remarkable proliferation inhibiting effect on Tca8113 cells. After being exposed to exogenous NO, Tca8113 cells showed series of apoptotic morphological changes such as cell shrinkage, nuclear condensation; and also showed DNA fragmentation, G2/M phase arrest as well as upregulation of the tumor suppressor P53 protein. CONCLUSION: Exogenous NO has a proliferation inhibition and apoptosis induction effect on Tca8113 cells in a concentration and time-dependent manner, P53 protein may be involved in the apoptosis induced by NO. PMID- 16470922 TI - [Effects of IGF-I and BMP-2 combined application on promoting proliferation, differentiation and calcification of MC 3T3-E1 and NIH 3T3 cells]. AB - OBJECTIVE: To demonstrate the effects of recombinant human insulin-like growth factor-I (rhIGF-I) and/or recombinant human bone morphogenetic protein-2 (rhBMP 2) on proliferation, differentiation and calcification of MC 3T3-E1 cells and NIH 3T3 cells. METHODS: Mouse osteoblast-like cell line MC 3T3-E1 and mouse fibroblast cell line NIH 3T3 were treated with different dosages of rhIGF-I or rhBMP-2 and rhIGF-I plus rhBMP-2. Cell proliferation was measured by methylthiazol tetrazolium (MTT)method and flow cytometry. Cell differentiation was examined by using alkaline phosphatase(ALP)measurement kit. Radioimmunoassay was applied to detect levels of osteocalcin (OC) secreted by cultured cells. Von kossa staining method was used to study the calcification effects. RESULTS: MC 3T3-E1 cells treated with 1-50 ng/ml rhIGF-I and NIH 3T3 cells treated with 5-75 ng/ml rhIGF-I showed marked effects of promoting proliferation (P<0.01), increasing the percentages of S-phase cells, decreasing the percentages of G1 phase cells and increasing activities of cellular ALP and percentages of calcification area (P<0.05). 10-100 ng/ml rhBMP-2 was also able to promote proliferation (P<0.01), increase the percentages of S-phase cells, decrease the percentages of G1-phase cells and enhancing cellular ALP activities and percentages of calcification area for both the two cells (P>0.05). CONCLUSION: rhIGF-I and rhBMP-2 have synergistical effects on promoting cell proliferation, early cell differentiation and calcification depending on the used dosages, but no significant effects on promoting advanced cell differentiation. PMID- 16470923 TI - [Nonalcoholic steatohepatitis in obese children: the prevalence and possible mechanism]. AB - OBJECTIVE: To investigate the prevalence of nonalcoholic steatohepatitis (NASH) in obese children and its possible mechanism. METHODS: Three subgroups were classified according to their body mass index (BMI) in 123 obese children with BMI over 23 aged 7 to 16:34 cases with BMI> or =31 group; 57 cases with 25< or =BMI<30 group; 32 cases with 23< or =BMI<25 group. Ultrasonographic and biochemical parameters including serum alanine aminotransferase (ALT), serum aspartate aminotransferase (AST), serum cholesterol, serum triglyceride, serum uric acid and free glucose to free insulin ratio (FGIR) were assayed. Twenty four children suspected as benign acanthosis nigricans underwent skin biopsy and its association with nonalcoholic steatohepatitis was also discussed. RESULTS: Ninety nine children (80.49 %) showed abnormal hepatic sonograms and 54 were diagnosed as NASH with the prevalence of 43.90%. Compared with the other two groups, BMI> or =31 group was significantly higher in prevalence of abnormal hepatic sonograms, NASH, decreased FGIR and risk of benign acanthosis nigricans (P<0.01). Fifty-four children diagnosed as NASH showed significantly higher incidence of hyperlipidemia, insulin resistance and higher body mass index as compared with 24 subjects without fatty liver changes. In 54 NASH children, 20(37.04%) had benign acanthosis nigricans. By bivariate analysis, ALT and AST were correlated well with BMI, cholesterol, triglyceride and FGIR (r(s)=0.413, 0.290, 0.379, -0.477, P<0.01; r(s)=0.359, 0.349, 0.348, -0.369, P<0.01). CONCLUSION: There is a high prevalence of nonalcoholic steatohepatitis in simple obese children and high incidence of benign acanthosis nigricans in NASH subjects. BMI> or=30 is a high risk factor of being NASH. Severe disturbance of lipid metabolism and insulin resistance may be involved in the mechanism of NASH. PMID- 16470924 TI - [Viraemia and extraintestinal involvement after rotavirus infection]. AB - OBJECTIVE: To study the incidence of viraemia and extraintestinal organ damage in children with acute rotavirus (RV) gastroenteritis. METHODS: Eighty-three children with acute rotavirus gastroenteritis were hospitalized from October 2002 to March 2003, whose blood and fecal samples were obtained on admission. Rotavirus RNA (encoding the VP7 outer capsid protein) were detected in blood and fecal samples by nest reverse transcription-polymerase chain reaction (RT-PCR). According to the result of blood RV-RNA, the patients were divided into RV-RNA positive group and RV-RNA negative group. The differences between these two groups in the severity of gastroenteritis and extraintestinal organ damage were analyzed. RESULTS: Eighty-two of 83 stool samples from the children with rotavirus infection were positive for rotavirus RNA. Sixteen of 83 blood samples were positive for rotavirus RNA with a positive rate of 19.3%. The nucleotide sequence of cloned cDNAs, resembling part of the VP7 gene, was identical from paired blood and fecal samples. There were no significant differences between blood RV-RNA positive group and blood RV-RNA negative group in the rate and degree of fever, diarrhea, dehydration, metabolic acidosis, hypokalemia and myocardial damage (P>0.05); while the incidences of liver damage, rash, lower respiratory tract infection and the central nervous system involvement in the blood RV-RNA positive group were significantly higher than those in the blood RV RNA negative group (P<0.05). CONCLUSION: Viraemia is present in the children with acute rotavirus gastroenteritis. Viraemia might be an important mechanism by which rotavirus spread to the extraintestinal sites resulting in organs damage. PMID- 16470925 TI - [Genotypes of aminoglycoside-modifying enzyme and clinical study of high-level gentamycin resistant enterococcus]. AB - OBJECTIVE: To determine the antibiotics resistance, aminoglycoside-modifying enzymes and homology of high-level gentamycin resistant enterococcus in clinical specimens. METHODS: The high-level gentamicin resistant (HLGR) isolates were screened by the agar method and the resistance of 14 antimicrobial agents was determined by K-B method. The aminoglycoside-modifying enzyme genes were detected by polymerase chain reaction (PCR). Pulsed-field gel electrophoresis (PFGE) was used to analyze the homology of HLGR isolates. RESULTS: The ratio of HLGR was 64.2% (68/106). Among the HLGR,there were no isolates resistant to linezolid, vancomycin and tecoplanin, and Enterococcus faecium was more resistant to beta lactam antibiotics and quinolone than Enterococcus faecalis. The positive rate of aac(6')-Ie-aph(2')-Ia was 92.6% and 3 isolates had the resistance gene mostly similar to aph(2')-Id. And among 51 HLGR isolates from the hospitalized patients, PFGE grouped 17 E. faecalis isolates into 4 clusters (A-D), and 33 E. faecium isolates into 8 clusters (A-H) with A cluster as predominant. CONCLUSION: HLGR has become the important antibiotic resistance bacteria which results in nosocomial infection; and aac(6')-Ie-aph(2')-Ia is the main aminoglycoside modifying enzyme gene which causes HLGR. PMID- 16470926 TI - [Evaluation of aminoglycoside resistance phenotypes and genotyping of acetyltransferase in Escherichia coli]. AB - OBJECTIVE: To investigate the prevalence of aminoglycoside resistance and genotyping of acetyltransferase in Escherichia coli. METHODS: Resistance phenotypes to 12 antibiotics of 44 Escherichia coli isolates were analyzed using agar dilution method and 3 aminoglycoside resistance genes aac(3)-I, II and aac(6')-I were determined by PCR method. RESULTS: In 44 clinical isolates, the occurrence of ESBLs was 45.45%, resistance rates were discrepant for amikacin (18.18%), gentamicin (56.82%) and tobramycin (61.36%), the prevalence of phenotype TG (tobramycin and gentamicin) indicative of aac(3)-II production and TGA (tobramycin, gentamicin and amikacin) indicative of aac(6')-I production were 36.36% and 18.18%, respectively. The most common aminoglycoside resistance genotype of acetyltransferase was aac(3)-II (52.27%) and aac(6')-I was lower (29.55%), but no aac(3)-I was detected. CONCLUSION: At least 2 acetyltransferase genes exist in this area i.e. aac(3)-II and aac(6')-I. PMID- 16470927 TI - [Determination of propofol in cerebral spinal fluid by HPLC with fluorescence detection]. AB - OBJECTIVE: To establish a method for determining propofol in human cerebrospinal fluid (CSF). METHODS: Reverse phase high-performance liquid chromatography (HPLC) with fluorescence detection was applied to quantitative analysis. CSF samples were centrifuged (12,500 r/min for 3 min) and filtered (the diameter of the filter is 0.45 microm). Twenty mul of supernatant was directly injected and separated by Supelco Discovery C(18)column. The mobile phase was composed of methanol-water (80:20); the flow rate was 1 ml/min, and the column temperature was 30 degree. The fluorescence detective waves were: lambda ex=276 nm and lambda em=310 nm. RESULTS: The linear range of propofol in CSF was 5-200 ng/ml (r=0.9994). The recovery rates for high, intermediate and low concentrations were 101.2%, 99.8%, 98.8%, respectively. The RSD of inter-day assay was 1.55%, 1.73%, 6.01% and it of intra-day assay was 1.69%, 2.37%, 8.60%. The limit of detection proved to be 2 ng/ml. CONCLUSION: The method is rapid, simple, accurate and sensitive for measurement of propofol concentration in CSF. PMID- 16470928 TI - [Relation between mutant p53 and multidrug resistance in gastric cancer]. AB - OBJECTIVE: To explore the relationship between mutant p53 and multidrug resistance in gastric cancer. METHODS: Mutant p53 (mp53) and mp53+sv40Tag were transferred to gastric cancer cell line SGC-7901. The MDR-1 mRNA was examined using RT-PCR, and the difference in chemotherapeutic sensitivity of SGC-7901 cells with mutant p53 was compared with those with mp53+sv40Tag and controls by MTT method. RESULTS: SGC-7901 cells with mutant p53 showed higher MDR-1 mRNA than that of other two groups. SGC-7901 cells with mutant p53 showed higher chemotherapeutic sensitivity to 5-Fu than that with mp53+sv40Tag and control (P<0.05), but no difference between those with mp53+sv40Tag and control (P>0.05). SGC-7901 cells with mutant p53 and those with mp53+sv40Tag showed higher chemotherapeutic sensitivity to ADM than control (P<0.05), but no difference between those with mp53 and with mp53+sv40Tag (P>0.05). There was no difference in chemotherapeutic sensitivity of SGC-7901 cells with mutant p53 compared with those with mp53+sv40Tag and control to CDDP (P>0.05). CONCLUSION: Mutante p53 genes relates to multidrug resistance of gastric cancer. PMID- 16470929 TI - [Enucleation of benign ovarian cysts by laparoscopy]. AB - OBJECTIVE: To evaluate the efficacy of improved laparoscopic enucleation of benign ovarian cysts. METHODS: A total of 234 cases of ovarian cysts with 271 cysts were analyzed retrospectively. 152 patients with 177 ovarian cysts (Group A) underwent the improved laparoscopic enucleation and 82 patients with 94 ovarian cysts (Group B) underwent the classic laparoscopic enucleation. The data of operative process and postoperative follow-up were compared between two groups. RESULTS: The rate of spillage of the Group A and Group B was 1.7% and 18.1% (P<0.01), respectively. The operating time was (40 +/-14)min and (47 +/- 16)min (P<0.01), respectively. The blood loss was (25 +/-17)ml and (27 +/- 19)ml (P>0.05), respectively. The bowel deflation recovery time was (18 +/- 8)h and (19 +/- 8)h (P>0.05), respectively. The length of hospital stay was (2.0 +/- 0.5)d and (2.2 +/- 0.8)d (P>0.05), respectively. CONCLUSION: Compared with classic laparoscopic procedure, the improved laparoscopic ovarian enucleation seems to be safer and more effective with shorter operating time. PMID- 16470930 TI - [MRI diagnosis of benign sacrococcygeal teratomas of the infants]. AB - OBJECTIVE: To investigate the value of MR imaging in the diagnosis of benign sacrococcygeal teratomas of the infants. METHODS: MR imaging of benign sacrococcygeal teratomas in 6 cases proved by surgery and pathology was retrospectively reviewed. In all patients, a fast imaging sequence, fast spin echo sequence was employed, together with short time inversion recovery sequence and contrast enhancement scanning by intravenous injection of Gd-DTPA. RESULTS: There were 6 benign sacrococcygeal teratomas, which were heterogeneous masses and arose from the distal sacrococcygeal region in the pelvis. The MR imaging appearances of the benign sacrococcygeal teratomas were characteristic, T1- and T2-weighted images demonstrated a large mass containing round, well-defined areas of varying signal intensity representing its cystic, solid, and sometimes fat, calcification within the lesions. CONCLUSION: MR imaging provides definitive information of benign sacrococcygeal teratomas and clearly shows both extra-and intra-pelvic components, and even better anatomic details, which facilitates the surgical planning adequately. PMID- 16470931 TI - [Research progress on human embryonic stem cell lines]. PMID- 16470932 TI - [Ruptured aneurysm of the sinus of Valsalva and ventricular septal defect complicating infective endocarditis: a case report]. PMID- 16470933 TI - More evidence concerning aspirin use before heart surgery. PMID- 16470934 TI - Preserving vision. Recognizing retinal problems early. PMID- 16470935 TI - Should people who are dark-skinned wear sunscreen? My husband, who is black, insists it's not necessary for him to use sunscreen. PMID- 16470936 TI - Sex and the heart - the correlation between sexual dysfunction and cardiovascular disease. Proceedings of a meeting. April 23, 2005. Galveston, Texas, USA. PMID- 16470938 TI - Some historical ruminations on the ACS Colloid and Surface Chemistry Symposium. PMID- 16470939 TI - Ryan White, ADAP, Medicaid funding problems could push HIV providers to the end of their tethers. Health care providers subsidize patchwork system. AB - AIDS advocates and HIV providers increasingly are pessimistic about the future of HIV patients in this nation when federal adn state cuts are forcing many more people on drug waiting lists and drug company compassionate care rolls. The problem is the medical care of the HIV population, even at its most efficient, cost far more than what any government and insurance funds will pay for it, and the situation is only getting worse, they say. PMID- 16470940 TI - TennCare once was model for public health care, but now is cautionary tale. Over 1,000 HIV patients left with no coverage. AB - TennCare's chief features of expanding health care coverage to uninsured and uninsurable people have been eliminated, and many HIV patients will lose their health care coverage, Stephen Raffanti, MD, MPH, chief medical officer of Comprehensive Care Center in Nashville, TN. PMID- 16470941 TI - CDC's most recent data on HIV epidemic now include stats from New York state. Overall picture shows little has changed recently. AB - The most interesting piece of the new HIV epidemic data coming from the Centers for Disease Control and Prevention (CDC) of Atlanta, GA, is that it includes statistics from New York state for the first time, which means that it finally provides a more comprehensive national picture. PMID- 16470942 TI - Homeless HIV program uses creativity, myriad of approaches to improve adherence. Program even has staff at race track. AB - HIV clinicians and researchers routinely note that one of the most difficult populations with which to achieve HIV antiretroviral adherence is the homeless. PMID- 16470937 TI - Comparative gene expression analysis by differential clustering approach: application to the Candida albicans transcription program. AB - Differences in gene expression underlie many of the phenotypic variations between related organisms, yet approaches to characterize such differences on a genome wide scale are not well developed. Here, we introduce the "differential clustering algorithm" for revealing conserved and diverged co-expression patterns. Our approach is applied at different levels of organization, ranging from pair-wise correlations within specific groups of functionally linked genes, to higher-order correlations between such groups. Using the differential clustering algorithm, we systematically compared the transcription program of the fungal pathogen Candida albicans with that of the model organism Saccharomyces cerevisiae. Many of the identified differences are related to the differential requirement for mitochondrial function in the two yeasts. Distinct regulation patterns of cell cycle genes and of amino acid metabolic genes were also revealed and, in some cases, could be linked to the differential appearance of cis regulatory elements in the gene promoter regions. Our study provides a comprehensive framework for comparative gene expression analysis and a rich source of hypotheses for uncharacterized open reading frames and putative cis regulatory elements in C. albicans. PMID- 16470944 TI - FDA notifications. Proposed changes to condom regulations. PMID- 16470943 TI - MMWR report on HIV and TB highlights challenges, needs. AB - In this Q&A story, Kevin Cain, MD, answers questions about the report on "Screening HIV-Infected Persons for Tuberculosis--Cambodia, January 2004-February 2005. PMID- 16470945 TI - FDA notifications. FDA postpones CCR5 Antagonist Review Group meeting. PMID- 16470946 TI - World AIDS Day comments by FDA's HIV/AIDS program director. PMID- 16470947 TI - Study of HIV risk in Africa shows women have very low risk behaviors. Even married women were at risk. PMID- 16470948 TI - [Diabetes mellitus: types 1 & 2 increasingly similar]. PMID- 16470949 TI - [Reducing obesity]. PMID- 16470950 TI - Trastuzumab in breast cancer. PMID- 16470951 TI - Trastuzumab in breast cancer. PMID- 16470952 TI - Trastuzumab in breast cancer. PMID- 16470953 TI - Trastuzumab in breast cancer. PMID- 16470954 TI - Long-acting methods of contraception. PMID- 16470955 TI - Medical management of depression. PMID- 16470957 TI - [Proceeding of 35th annual meeting of the Japanese Society of Neuropsychopharmacology. Osaka, Japan. July 6-8, 2005. Abstracts]. PMID- 16470956 TI - Medical management of depression. PMID- 16470958 TI - [The 47th Congress of the Japanese Society of Clinical Hematology. Yokohama, Japan. September 17-19, 2005. Program and Abstracts]. PMID- 16470959 TI - Social inference and mortuary practices: an experiment in numerical classification. PMID- 16470960 TI - Popular movements and late Roman cemeteries. PMID- 16470961 TI - Mary Abbott Hess, LHD, MS, RD, FADA, receives 2005 Copher Memorial Award. PMID- 16470962 TI - A classic Maya family tomb. PMID- 16470963 TI - Mortality of the oldest old in China. AB - OBJECTIVE: This study investigates the role of customary activities, both social and solitary, in mortality among the oldest old in China. METHODS: The data come from the Chinese Longitudinal Healthy Longevity Survey. Weibull hazard models analyze the mortality risk of those 80 years of age and older within a 2-year period between 1998 and 2000. RESULTS: Results show that solitary activities, either active or sedentary, are significantly associated with lower mortality risk. The effect of social activities on mortality gradually diminishes with age and is reversed at very old ages when physical exercise, health status, and sociodemographic characteristics are controlled. DISCUSSION: Customary activities, which are less physically demanding, show independent effects on the elderly's survival. Withdrawal from social contacts may be an adaptive response to challenges faced at very advanced ages. It is important to recognize the unique characteristics of this rapidly growing population. PMID- 16470964 TI - Amino acid racemization dating of fossil bones. PMID- 16470965 TI - Advances and results in radiocarbon dating: early man in America. PMID- 16470966 TI - Age differences in the correlates of physical functioning in patients with chronic pain. AB - OBJECTIVES: Pain-related disability in the elderly is a major contributor to poor quality of life. In this study, the author examined age-related differences in the factors associated with physical disability in the context of chronic pain. METHOD: A large cohort of chronic pain patients was categorized into four age groups, and correlates of physical disability were evaluated within each age category. RESULTS: Several important age differences in the predictors of physical disability were noted. Affective distress was unrelated to physical disability among the elderly, whereas this association was quite strong among younger patients. In contrast, pain severity was a weak predictor of disability within the younger group, but it accounted for more than one fourth of the variance in physical disability in those older than 65. DISCUSSION: These data suggest that the determinants of functioning may vary across the life span, implying a potential for pain management programs tailored to a patient's age. PMID- 16470967 TI - Biology, biogeography and man. PMID- 16470968 TI - Are managed care Medicare beneficiaries with chronic conditions satisfied with their care? AB - OBJECTIVES: This article compares patient experiences of chronically ill older people in health maintenance organizations (HMOs) with other forms of Medicare supplemental coverage. METHOD: Using data from the 1996 Medicare Current Beneficiaries Survey, the authors analyzed the experiences of chronically ill elderly with overall quality, access to care, and physicians' technical, interpersonal, and information-giving skills. Logistic models controlled for prevalent chronic conditions, functioning, perceived health status, sociodemographics, region of residence, and county-level Medicare HMO penetration. RESULTS: Satisfaction with quality of overall care and physicians' skills was more likely for many conditions for those with private fee for service and Medicaid supplemental coverage, compared to Medicare HMO population. No insurance effects were found among elders who had none of the examined conditions. DISCUSSION: Managed care may have negatively affected patients' perceptions of overall quality of care and doctor-patient interaction. Including additional and supplementary services to the delivery of care may improve satisfaction rates. PMID- 16470969 TI - Colonization cycles in man and beast. PMID- 16470970 TI - Long term hunter-gatherer adaptation to desert environments: a biogeographical perspective. PMID- 16470971 TI - Biogeographical theory and cultural analogies. PMID- 16470972 TI - Evolution is no help. PMID- 16470973 TI - Geographic systems and human diversity in the North Solomons. PMID- 16470974 TI - Recommendations for the supervision of pathology assistants. PMID- 16470975 TI - Terminological problems in quantitative faunal analysis. PMID- 16470976 TI - Teotihuacan regional population administration in eastern Morelos. PMID- 16470977 TI - The effects of resistance training and walking on functional fitness in advanced old age. AB - The authors assessed the effects of resistance training and walking exercise on measures of functional fitness. Sixty-four volunteers (average age 83.5 years) from an independent-living facility were randomly assigned to walking, resistance training, or control groups. Participants in the walking and resistance-training groups engaged in two exercise sessions per week for 16 weeks. Measures of functional fitness included upper and lower body strength, hip and shoulder flexibility, agility and balance, coordination, blood pressure, and resting heart rate. Repeated measures analysis of variance was used to examine pretest to posttest differences. Both exercise groups showed significant improvements relative to control group in upper and lower body strength, shoulder flexibility, and agility and balance exercise. Findings demonstrate that exercise can lead to improvements in multiple domains of functional fitness even among very old, previously sedentary individuals, possibly making activities of daily living easier to perform. PMID- 16470979 TI - Chronic diseases a major cost. PMID- 16470978 TI - Medicine prices and a medicine price index. PMID- 16470980 TI - Stop press--Beware of 'overpayment sting'. PMID- 16470981 TI - Racial and ethnic differences in use of assistive devices for mobility: effect modification by age. AB - PURPOSE: The authors investigated the extent to which differences in mobility device use existed by race/ethnicity and whether the effects were modified by age. METHOD: Using Phase II of the National Health Interview Survey on Disability, the authors identified 7,148 mobility-impaired adults. Logistic regression models provided estimates of race/ethnicity, adjusting for predictors of device use. RESULTS: Blacks were 1.2 times as likely as non-Hispanic Whites (95% CI: 1.0-1.4) to use devices. Hispanics were 0.78 times less likely to use devices as were non-Hispanic Whites (95% CI 0.65-0.96). Differences were attenuated by age, with a 40% increased likelihood of use attributable to being Black and aged 45 to 75, a 30% increased likelihood attributable to being Hispanic and aged 65 to 75, and a 130% increased likelihood attributable to being Hispanic and older than 75. CONCLUSION: Disentangling the observed patterns in mobility device use will provide direction for development of interventions to promote assistive device use. PMID- 16470982 TI - Being healthy is not enough when it comes to risk cover. PMID- 16470983 TI - Food: tradition and change in Hellenistic Egypt. PMID- 16470984 TI - Diet and dental disease. PMID- 16470985 TI - Prehistoric diet and subsistence of the Moche Valley, Peru. PMID- 16470986 TI - Diet, nutrition and population dynamics in the basin of Mexico. PMID- 16470987 TI - Initial perspectives on prehistoric subsistence in the inland Niger Delta (Mali). PMID- 16470988 TI - Kinship interrelations in a fifth-century 'Pannonian' cemetery: an archaeological and palaeobiological sketch of the population fragment buried in the Mozs Cemetery, Hungary. PMID- 16470989 TI - Hierarchy levels, sum score, and worsening of disability are related to depressive symptoms in elderly men from three European countries. AB - OBJECTIVES: The objectives were to investigate the predictive value of hierarchy levels and sum score of disability and change in disability on depressive symptoms. METHOD: Longitudinal data of 723 men age 70 and older from the Finland, Italy, and the Netherlands Elderly Study were collected in 1990 and 1995. Self reported disability was based on three disability domains (instrumental activities, mobility, and basic activities) and depressive symptoms on the Zung questionnaire. RESULTS: Severity levels of disability were positively associated with depressive symptoms. Men with no disability scored 5 to 17 points lower (p < .01) on depressive symptoms than did those with disability in all domains. Among men with mild disability, those who had worsening of disability status in the preceding 5 years scored 5 points higher (p = .004) on depressive symptoms than did men who improved. DISCUSSION: Hierarchic severity levels, sum score of disability, and preceding changes in disability status are risk factors for depressive symptoms. PMID- 16470990 TI - Piaget, stone tools and the evolution of human intelligence. PMID- 16470991 TI - A time of change. PMID- 16470992 TI - Plant foods in savanna environments: a preliminary report of tubers eaten by the Hadza of Northern Tanzania. PMID- 16470993 TI - Early Holocene mortuary practices and hunter-gatherer adaptations in southern Somalia. PMID- 16470995 TI - Water supply and sewage disposal at Mohenjo-Daro. PMID- 16470994 TI - Social, spatial and chronological patterning in marine food use as determined by delta13C measurements of Holocene human skeletons from the south-western Cape, South Africa. PMID- 16470996 TI - Prehistoric health status of the Roonka population. PMID- 16470997 TI - Infant and childhood morbidity and mortality risks in archaeological populations. PMID- 16470998 TI - The archaeology of public health at York, England. PMID- 16470999 TI - The palaeopathology of leprosy in Britain: a review. PMID- 16471000 TI - The cemetery of the Hospital of St James and St Mary Magdalene, Chichester - a case study. PMID- 16471001 TI - Public health and private sentiment: the development of cemetery architecture and funerary monuments from the eighteenth century onwards. PMID- 16471002 TI - Living on the Boott: health and well being in a boardinghouse population. PMID- 16471003 TI - Carotid endarterectomy. PMID- 16471004 TI - Vascular neurology. PMID- 16471005 TI - Consultation on Human Fertilisation and Embryology Act. AB - Following the review of the Human Fertilisation and Embryology Act by the House of Commons select committee on science and technology (see Bulletin 208), the Department of Health issued a consultation paper on its proposals to change the law. Although the consultation period will be over before this issue is published, it seems worth reprinting the summary of the paper to record the department's thinking on a variety of issues. PMID- 16471006 TI - Evidence of prehistoric diet from northern Chile: coprolites, gut contents and flotation samples from the Tulan Quebrada. PMID- 16471007 TI - Complications within consent. AB - The consent process is intended to protect patients, prevent complaint and litigation, and help practitioners to be accountable, to justify their decisions, and to inform and involve patients/parents in their health care. Medico-legal guidance poses uncertainties and dilemmas that can undermine these aims and deter some practitioners from informing and involving patients/parents in decision making. Greater recognition of contradictions and limitations in the guidance on consent could assist practitioners and patients/parents in their shared decision making. PMID- 16471008 TI - A chronological framework for human origins. PMID- 16471009 TI - Palaeolithic chronology and possible coexistence of Homo erectus and Homo sapiens in China. PMID- 16471010 TI - Revolution or resolution? The archaeology of modern human origins. PMID- 16471011 TI - An overview of the major changes in the 2002 APA Ethics Code. AB - This article summarizes the major changes that were made to the 2002 Ethical Principles and Code of Conduct of the American Psychological Association. The 2002 Ethics Code retains the general format of the 1992 Ethics Code and does not radically alter the obligations of psychologists. One goal of the Ethics Committee Task Force was to reduce the potential of the Ethics Code to be used to unnecessarily punish psychologists. In addition, the revised Ethics Code expresses greater sensitivity to the needs of cultural and linguistic minorities and students. Shortcomings of the 2002 Ethics Code are discussed. PMID- 16471012 TI - Human biology, environment and ritual at Khok Phanom Di. PMID- 16471013 TI - Bioinformatics challenges in proteomics. AB - A little after the genomic revolution had been celebrated, it seemed as if a competition began to found new -omics disciplines that ultimately all have the same goal, the understanding of biological function. There are many similar definitions for proteomics that can be summarized as follows: proteomics is a large-scale study of structure and function of proteins in an organism or cell. Importantly, the proteome is much more variable than the genome through its interactions with the genome and secondary modifications. It differs depending on the tissue and stage in life-cycle. Hence, proteomics is a very diverse discipline that uses a variety of experimental set-ups and targets in order to elucidate function. Its dissociation from other disciplines can only remain artificial. The bioinformatics applied to proteomics are equally varied. In this review we will focus mainly on a few areas of bioinformatics that seem to us as particularly noteworthy or characteristic for proteomics research, for example in 2DE analysis or mass spectrometry. Another important task of bioinformatics is the prediction of functional properties. We will summarize the approaches taken in order to predict protein networks, which are based on the extensive integration of several kinds of -omics data. We will give a short overview of a demanding field in computational biology, the analysis and prediction of protein 3D structures. In order to provide a broader perspective we will close this review with a generalized description of activities and databases in the realm of proteomics. PMID- 16471014 TI - The archaeobotany of subsistence in the Pacific. PMID- 16471015 TI - The Schiavo case and the culture wars. PMID- 16471016 TI - Annihilating Terri Schiavo. PMID- 16471017 TI - John Paul II and America's laws on life. PMID- 16471019 TI - Teaching without harming the living: performing minimally invasive procedures on the newly dead. PMID- 16471018 TI - The least of my brethren: the ethics of heterologous embryo transfer. PMID- 16471020 TI - Pelvic examinations under anesthesia: an important teaching tool. PMID- 16471021 TI - Molecular biology and archaeology: a prospectus for inter-disciplinary research. PMID- 16471022 TI - Archaeology and genetics: analysing DNA from skeletal remains. PMID- 16471023 TI - Autonomy suspended: using female patients to teach intimate exams without their knowledge or consent. PMID- 16471024 TI - Blood residues on stone tools: indoor and outdoor experiments. PMID- 16471025 TI - The artifact as site: an example of the biomolecular analysis of organic residues on prehistoric tools. PMID- 16471026 TI - Research involving children: regulations, review boards and reform. PMID- 16471027 TI - Biomolecular archaeology of wheat: past, present and future. PMID- 16471028 TI - Do pregnant women have (living) will? PMID- 16471029 TI - Biomolecular archaeology and lipids. PMID- 16471030 TI - Identifying problematic remains of ancient plant foods: a comparison of the role of chemical, histological and morphological criteria. PMID- 16471031 TI - Is the human zygote a person? PMID- 16471032 TI - Stem cell research, cloning and Catholic moral theology. PMID- 16471033 TI - The brave new world of embryonic stem cell research: utilitarian consequentialism and faulty moral reasoning. PMID- 16471034 TI - Democracy and abortion. PMID- 16471035 TI - Chemosensitization of hormone-refractory prostate cancer cells by curcumin to TRAIL-induced apoptosis. AB - Failure to undergo apoptosis has been implicated in the resistance of tumor cells to anticancer therapies. Promotion of apoptosis in tumor cells could potentially increase the efficacy of conventional treatment regimens and improve prognosis. Prostate cancer cells are generally resistant to induction of apoptosis by anticancer agents and death ligands. We investigated the sensitization of prostate cancer cell lines by curcumin (diferuloyl-methane) to TNF-related apoptosis inducing ligand (TRAIL)-induced apoptosis. Prostate cancer cells treated with curcumin or TRAIL or curcumin and TRAIL together were assessed for induction of apoptosis and pathway of apoptosis was determined from the activation of procaspases and release of cytochrome c from mitochondria. Curcumin sensitized LNCaP, DU145 and PC3 tumor cell lines to TRAIL. Combined curcumin and TRAIL treatment produced the most loss of viable cells by inducing apoptosis as revealed by accumulation of hypodiploid cells in sub-G1 phase, enhanced annexin V binding, DNA fragmentation, cleavage of procaspases-3, -8, and 9, truncation of proapoptotic Bid, and release of cytochrome c from mitochondria. Tumor cells expressed constitutively active NF-kappaB and sensitization to TRAIL involved inhibition of NF-kappaB by curcumin. These findings suggest that combined curcumin/TRAIL chemo-immunotherapy may be a beneficial adjunct to the standard therapeutic regimens for prostate cancer. PMID- 16471036 TI - A stem cell fusion model of carcinogenesis. AB - The origin of cancer remains enigmatic. Current models of carcinogenesis based on the gene mutation hypothesis have limitations in explaining many aspects of cancer. We put forward a new model of multistage carcinogenesis and propose that cancer development involves gene mutations and cell fusions. Specifically, cancer can result from a fusion between an "altered" pre-malignant cell and a bone marrow-derived stem cell (BMDSC). "Aneuploidy", which is a hallmark of malignancy, is a direct consequence of this cell fusion. The "stem cell fusion" model explains the remarkable similarities between malignant cells and BMDSC. This model also explains why non-mutagens can be carcinogens, and why non mutagenic processes, such as wound healing and chronic inflammation, can promote malignant transformation. This model is readily testable. Cancer has been difficult to treat because of tissue heterogeneity and gene instability. However, if the malignant characteristics of cancer cells are derived from BMDSC, new conserved targets such as homing receptors for designing novel therapies may emerge. PMID- 16471037 TI - Selection of cell lines with enhanced invasive phenotype from xenografts of the human prostate cancer cell line WPE1-NB26. AB - Prostate cancer is a leading cause of death from cancer in American men and metastasis the main cause of death. To better understand the disease and accelerate development of new therapies, in vivo models that reflect different disease stages are needed. A family of cell lines that mimics multiple steps in cancer development and tumor progression has been developed in our laboratory from the parent, non-tumorigenic, RWPE-1 cell line by transformation with N methyl-N-nitrosourea (MNU). The MNU cell lines mimic multiple steps in tumor progression where WPE1-NB26 is the most malignant cell line. WPE1-NB26 cells form metastases in the lungs of athymic, male, nude mice after intravenous injection. Two new cell lines, WPE1-NB26-64 and WPE1-NB26-65, showing more malignant characteristics than the parent WPE1-NB26 cell line, were derived from tumors after subcutaneous injection of WPE1-NB26 cells into nude mice. The WPE1-NB26-64 and WPE1-NB26-65 cell lines show an increase in anchorage-dependent growth and invasive ability as compared to the parent WPE1-NB26 cells. While the parent WPE1 NB26 cells express barely detectable levels, the new cell lines produce high levels of matrix metalloproteinase MMP-2 and detectable levels of MMP-9. By immunostaining, all three cell lines were positive for cytokeratins CK18 and CK5/14. These cell lines, having the same lineage, represent additional steps in the multi-step process of tumor progression and provide novel and useful cell models for studies on tumor progression and for drug development for the treatment of prostate cancer. PMID- 16471038 TI - Clinical role of determination of plasma mitotane and its metabolites levels in patients with adrenal cancer: results of a long-term follow-up. AB - Our study aimed at evaluation of the relations between the plasma levels of mitotane (o,p'-DDD) and its metabolites, o,p'-DDA and o,p'-DDE, and the efficacy of Mitotane therapy during a long-term follow-up. Eighteen patients, aged 11 to 70 years, were included to the study. Metastatic or regional stage was diagnosed in 15 patients, while localized disease in three patients. Mitotane has been administered in daily doses of 3.0 to 10.0 g in metastatic and regional stages, and 1.5 to 4.0 g in the localized disease, simultaneously with hydrocortisone, prednisolone and fludrocortisone. Mitotane and its metabolites were determined by a high-pressure liquid chromatographic method. The plasma o,p'-DDD level exceeding 44 _M/L, considered as curative one, was reached in nine cases. The o,p'-DDA/o,p'-DDD ratio rose significantly mainly in the first 1-3 months of therapy. The o,p'-DDE levels rose slowly, reaching higher values in long-term therapy, over 12 months of mitotane administration. In the group of patients with regional or metastatic stage, both the o,p'-DDE levels and the o,p'-DDE/o,p'-DDD ratios were higher in the survivors than in non-survivors. The results of our study suggest that the plasma concentrations of o,p'-DDE were more closely related to clinical improvement or remission than the o,p'-DDD levels. PMID- 16471039 TI - Expression patterns of fatty acid binding proteins in breast cancer cells. AB - We studied the expression levels of fatty acid binding proteins in breast normal and cancer cell lines. Liver fatty acid binding protein (L-FABP) and intestine fatty acid binding proteins were shown to be up-regulated in breast cancer cell lines while adipose- and epidermal-fatty acid binding proteins were down regulated in breast cancer cells compared to normal breast cell lines. We have previously shown that blocking the expression of L-FABP resulted in remarkable effects on apoptosis and cell proliferation of prostate cancer cell lines (Hammamieh et al 2004). To study the mechanism of effect of the liver fatty acid binding protein in breast cancer cells, we designed an antisense oligodeoxynucleotide to block the production of liver-FABP in MCF-7 cells. The antisense was shown to inhibit the expression of L-FABP and to induce apoptosis in MCF-7 cells. This study examines the mechanism by which L-FABP antisense regulates proliferation and apoptosis in breast cancer cell lines. We used human cDNA microarrays to explore differentially expressed genes in MCF-7 breast cancer cells treated with L-FABP antisense oligonucleotide. Some of the genes that were differentially expressed were confirmed using quantitative RT-PCR. Genes related to cell growth, proliferation and angiogenesis showed significant variations. This suggests a possible use of these antisense ODNs as therapeutic agents for breast cancer in future. PMID- 16471041 TI - Gene therapy in cancer: the missing point. AB - Over the last century cancer research has produced data leading to a composite picture where gene mutations and epigenetic phenomena strictly relate and overlap. This complexity has repercussions on the anti-cancer therapeutical strategies. The therapeutic pathway paved by the kochian one-cause/one disease principle fails in front of a multigenic multiphenomena disease like cancer. We still do not know what target(s) to hit/modify in order to prevent/stop the carcinogenic progression. On the light of cancer statistics 2005, we discuss the need of exactly defining the cancer targets in order to exploit the high potential of gene therapy. PMID- 16471040 TI - Cathepsin D expression in human colorectal cancer: relationship with tumour type and tissue differentiation grade. AB - OBJECTIVE: Cathepsin D (CD) is one of the main proteolytic enzymes contributing to the development of cancer. The aim of this study was to CD activity assay in the homogenates of tissues from the centre of the tumour (0) and tumour free area 2 cm, and 5 cm from the tumour border in human colorectal cancer. Activity in the centre of the tumour was compared with immunohistochemical expression CD. METHODS: CD activity was measured using acid denatured Hb as a substrate. For immunohistochemical staining peroxidase method was used. RESULTS: Activity of CD was significantly higher (15-fold) in tumour tissue homogenates in comparison to normal mucosa adjacent (control) (p < or = 0.0001) and raised parallel to the stage of tumour tissue differentiation grade. CD activity decreased significantly (p < or = 0.0001) with the distance from the tumour border 2 cm (12.7 fold) and 5 cm (5.7 fold) in comparison to the centre of the tumour. In immunohistochemical examinations CD was detected as diffuse cytoplasmic as well as fine granular staining of the cytoplasm, with occasional coarse cytoplasmic granules staining in the same cases that were positive for both. Positive staining was observed in 2 of 3 in well-differentiated (66%), 4 of 10 in moderately-differentiated (40%) and 4 of 5 in poorly-differentiated (80%), tubular adencarcinomas represented: 3 of 7 (42%) and 9 of 13 in invasive adencarcinoma (69%). CONCLUSION: We have observed a wide range of cathepsin D and their antigen expressions patterns in colorectal tumours with the development the disease stage, this finding may be used as a daignostic tumor marker in colorectal cancer. PMID- 16471042 TI - Developing end-of-life care for older people in care homes. PMID- 16471043 TI - Barriers to effective uptake of cancer screening among Black and minority ethnic groups. AB - AIM: To describe some of the factors that act as barriers to effective uptake of breast and cervical cancer screening services among black minority ethnic (BME) groups living in Brent and Harrow in the UK. DESIGN: A series of focus groups among African Caribbean, African, Gujarati, Pakistani, Greek and Arabic groups were held to discover their perceptions of cancer screening, the barriers to effective uptake and some strategies for intervention. SAMPLE: This consisted of 135 participants: 85 women and 50 men. RESULTS: Analysis of focus group data has revealed poor knowledge, underlying health and cultural beliefs, attitudes, language and unhelpful attitudes of health professionals to be important barriers. In terms of strategies for effective intervention, the most popular strategy for improving uptake of screening services was community-based cancer awareness education that is sensitive to religious and cultural needs. CONCLUSION: There is a need to provide community-based education to increase the uptake of screening services among BME groups. It is essential to plan concurrently to educate GPs and other health professionals in cultural beliefs and customs, language needs, racial awareness and communication skills. PMID- 16471045 TI - The law regarding assisted dying for the terminally ill in the UK. AB - This article outlines the Assisted Dying for the Terminally Ill Bill in the UK. The conditions that must be satisfied before an attending physician can assist a patient to die are highlighted and information regarding the declaration is provided. It is not certain that the Bill will obtain legislative approval and, if it does, there are statutory provisions for monitoring each assisted death. PMID- 16471044 TI - A guide to wound managment in palliative care. AB - Wound management in palliative patients is often a very challenging area of care. There are many unique issues that can combine to produce complicated wound management scenarios, including the types of wounds and wound symptoms most commonly affecting palliative care patients, as well as the presence of concurrent disease and associated treatment. Problems exist with the availability of suitable dressings and balancing life expectancy with the goals of wound care. A significant, and possibly under-recognized, issue is the emotional and social distress experienced by these patients, which can be directly attributed to their wound. These problems must all be recognized and addressed in order to manage wounds effectively in this patient population. This article aims to explore these issues and offer advice on the management of wound-related symptoms, with the ultimate goal of improving patients' quality of life. PMID- 16471046 TI - Introducing 'Palcall': an innovative out-of-hours telephone service led by hospice nurses. AB - Palcall is an out-of-hours telephone service providing support and advice to palliative care patients, their carers and health professionals. Each patient who is registered with the scheme consents to named friends or relatives having access to the dedicated Palcall number. Telephone calls to the service are taken by the most senior palliative care nurse on duty in the hospice. The nurse has ready access to details of the medical condition and current medications of every patient who is registered. If required, the nurse can contact a more senior nursing colleague or the on-call hospice GP. The day following the telephone call, the Palcall administrator forwards written details to the patient's GP and any healthcare professional involved in the care of the patient. Quality control measures are in place to continually enhance the service. PMID- 16471047 TI - Using computer-assisted learning to gain knowledge about child death and bereavement. AB - Computer-assisted learning (CAL) can be a useful tool in helping nurses to learn about child death and bereavement. A CAL package on the care of children with life-limited illnesses and their families has been designed, developed and evaluated with students on the diploma in nursing (child) programme. Early indications are that CAL, in conjunction with teacher support, can offer a means to allow individuals to work at their own pace, apply learning and explore new learning in a non-threatening environment. This article argues that the skills required to care holistically for children with life-threatening illnesses and their families are complex. Extensive applications of previously learnt transferable skills, as well as a good grasp of multidisciplinary working, are needed. In turn, these skills are transferable and applicable in almost any other professional situation. CAL combines different learning styles, such as factual information giving, enquiry-based and practice-based learning; therefore, integrating learning about child death and bereavement both in theory and in practice. PMID- 16471048 TI - Information at your fingertips: making the most of the Internet. PMID- 16471049 TI - A hardwon dirhodium paddlewheel with guanidinate type (hpp) bridging ligands. AB - We report a dirhodium paddlewheel compound which has an Rh2(5+) core enveloped by four guanidinate ligands and an axially coordinated bromine atom; Rh2(hpp)4Br crystallizes as a 1-D chain with a Rh-Rh distance of 2.430(3) e and a Rh-Rh bond order of 1.5 (hpp = the anion of 1,3,4,6,7,8-hexahydro-2H pyrimido[1,2a]pyrimidine). PMID- 16471050 TI - Construction of an interpenetrated 3-D network by [2 + 2] cycloaddition of Zn(ClO4)2 and 1,2-trans-(4-pyridyl)ethene: counter anion effects. AB - The reaction of Zn(ClO)2 with 1,2-trans-(4-pyridyl)ethene (bpe) results in an interpenetrated network containing both the starting bpe ligand and the newly generated 1,2,3,4-tetrakis(4-pyridyl)cyclobutane (4pycb) (formed by [2 + 2] cyclization of the starting ligand), while the reactions of Zn(NO3)2 and ZnSO4 with bpe result in a dinuclear complex and a two-dimensional sheet, respectively. PMID- 16471051 TI - Highly efficient binding of trivalent f-elements from acidic media with a C3 symmetric tripodal ligand containing diglycolamide arms. AB - Tripodal chelates bearing three diglycolamide units precisely arranged on a triphenoxymethane platform were synthesized to mimic the preferred tricapped trigonal prismatic geometry favored by lanthanides with oxygen donor ligands, and the ligand binds heavier lanthanides very efficiently in acidic media. PMID- 16471052 TI - Bis(isothiocyanato)bis(phosphine) complexes of group 10 metals: reactivity toward organic isocyanides. AB - Treatment of Ni(NCS)2(PMe2Ph)2 with organic isocyanides CN-R gave five-coordinate isocyanide Ni(II) complexes, Ni(CN-R)(NCS)2(PMe2Ph)2 (R = C6H3-2,6-Me2 (1), t-Bu (2)). Interestingly, the corresponding reaction of Ni(NCS)2(P(n-Pr)3)2 with 2 equiv. of CN-t-Bu gave an unusual compound, which exists as an ion pair of the trigonal bipyramidal cation [Ni(P(n-Pr)3)2(CN-t-Bu)3]2+ (3) and the dinuclear NCS bridged anion [Ni(1,3-micro-NCS)(NCS)3]2(2-) (4). In contrast, Pd(NCS)2(P(n Pr)3)2 underwent substitution with 2 equiv. of CN-t-Bu to give the four coordinate mono(isocyanide) Pd(II) complex Pd(NCS)(SCN)(CN-t-Bu)(P(n-Pr)3) (5) via phosphine dissociation. Reactions of M(NCS)2L2 (M = Pd, Pt; L = PMe3, PEt3, PMePh2, P(n-Pr)3) with two equiv. of CN-R (R = t-Bu, i-Pr, C6H3-2,6-Me2) gave the corresponding bis(isocyanide) complexes [M(CN-R)2(PR3)2](SCN)2 (7-13), except for Pd(NCS)2(PEt3)2 that reacted with CN-R' (R' = i-Pr, C6H3-2,6-Me2) and produced the mono(isocyanide) Pd(II) complexes [Pd(CN-R')(SCN)(PEt3)2](SCN) (14 and 15). Finally, treatment of M(NCS)2(PMe3)2 (M = Ni, Pd, Pt) with sterically bulky isocyanide CN-C6H3-2,6-i-Pr2 gave various products, (16-18) depending on the identity of the metal. PMID- 16471053 TI - Synthesis and photochemical characterization of a zinc phthalocyanine-zinc porphyrin heterotrimer and heterononamer. AB - The synthesis of two phthalocyanine-porphyrin covalently linked heteromolecules are described. Intramolecular energy transfer is investigated and quantified in terms of the quantum yield of energy transfer and found to be highly effective in both molecules. The photophysical properties of both molecules are modified greatly by the presence of the porphyrin moieties on the phthalocyanine core. PMID- 16471054 TI - Oxidative DNA damage induced by autoxidation of microquantities of S(IV) in the presence of Ni(II)-Gly-Gly-His. AB - NiIIGGH (GGH = glycylglycylhistidine) reacts rapidly with S(IV), in air-saturated solution, to produce NiIIIGGH. A mechanism is proposed where initial NiIII oxidizes SO3(2-) to SO3*-, which reacts with dissolved oxygen to produce SO5*-, initiating radical chain reactions. DNA strand breaks and 8-oxo-7,8-dihydro-2' deoxyguanosine formation were observed in air-saturated solutions containing micromolar concentrations of Ni(II) and S(IV). The extent of DNA damage showed dependence on the ratio of the NiIIGGH : S(IV) concentrations and the ionic strength. PMID- 16471055 TI - Polynuclear complexes with bridging pyrophosphate ligands: synthesis and characterisation of {[(bipy)Cu(H2O)(mu-P2O7)Na2(H2O)6] x 4H2O}, {[(bipy)Zn (H2O)(mu-P2O7)Zn(bipy)]2 x 14H2O} and {[(bipy)(VO)2]2(mu-P2O7)] x 5H2O}. AB - The reaction in water of M(II) ions (M = Cu, 1; Zn, 2; VO, 3) with 2,2' bipyridine (bipy) followed by Na4P2O7 leads to the formation of three new complexes which feature the pyrophosphate anion, P2O7(4-), as a bridging ligand. Single crystal X-ray diffraction revealed 1 to be {[(bipy)Cu(H2O)(micro P2O7)Na2(H2O)6] x 4H2O}, and 2 as a tetranuclear Zn(II) complex, {[(bipy)Zn(H2O)(micro-P2O7)Zn(bipy)]2 x 14H2O}. The structure of 1 consists of a mononuclear [(bipy)Cu(H2O)(P2O7)]2- unit that links via a pyrophosphate bridge to two Na atoms. The hydrated six-coordinate Na atoms themselves join together through bridging water molecules to generate a 2D Na-water sheet. The structure of 2 consists of a tetranuclear Zn(II) cluster (dimer-of-dimers) with two pyrophosphate ligands bridging between four metal centres. Adjacent clusters interact through face-to-face pi-pi interactions via the bipy ligands to yield a 2D sheet. Adjacent sheets pack in register to create channels, which are filled by the water molecules of crystallisation. An intricate 2D H-bonded water network separates adjacent sheets and encapsulates the tetranuclear clusters. Aspects of the pyrophosphate coordination modes in 1 and 2 are of structural relevance to those found within the inorganic pyrophosphatases. Compound 3, {[(bipy)(VO)2]2(micro-P2O7)] x 5H2O}, was isolated as an insoluble lime-green powder. Its dinuclear structure was elucidated from elemental and thermal analysis, magnetic susceptibility measurement and IR spectroscopy. The latter displayed characteristic bridging pyrophosphate and signature V=O stretches, which were corroborated by contrast to the IR spectra of 1 and 2 and through comparison with those found in the structurally characterised dinuclear complex, {[(bipy)Cu(H2O)]2(micro-P2O7) x 7H2O}, 4. PMID- 16471056 TI - Novel Ti-O-Ti bonding species constructed in a metal-oxide cluster. AB - The preparation and structural characterization of a novel Ti-O-Ti bonding complex constructed in the mono-lacunary alpha-Keggin polyoxometalate (POM), are described. The water-soluble, crystalline complex with a formula of K5H2[[{Ti(OH)(ox)}2(micro-O)](alpha-PW11O39)] x 13H2O 1 was prepared in 30.2% (0.60 g scale) yield in a 1 : 3 molar-ratio reaction of the tri-lacunary species of alpha-Keggin POM, Na9[A-PW9O34] x 19H2O, with the titanium(IV) source, K2TiO(ox)2 x 2H2O (H2ox = oxalic acid), in HCl-acidic solution (pH 0.08), and characterized by complete elemental analysis, thermogravimetric and differential thermal analyses (TG/DTA), FTIR, solution (31P, 183W, 1H and 13C) NMR spectroscopy and X-ray crystallography. The complex was also obtained in 47.6% (0.81 g scale) yield in a 1 : 2 molar-ratio reaction of the mono-lacunary Keggin POM, K7[PW11O39] x 10H2O, with the anionic titanium(IV) complex under acidic conditions. The molecular structure of [[{Ti(OH)(ox)}2(micro-O)](alpha-PW11O39)]7 1a, was successfully determined. This POM in the solid state is composed of one host (mono-lacunary site) and two guests (two octahedral Ti groups), in contrast to most titanium (IV)-substituted POMs consisting of one host and one guest. On the other hand, the 31P NMR measurements revealed that in aqueous solution this POM was present under a dissociation equilibrium which depends upon both temperature and pH. PMID- 16471057 TI - 2-[(8-Hydroxyquinolinyl)methylene]hydrazinecarboxamide: expanding the coordination sphere of 8-hydroxyquinoline for coordination of rare-earth metal(III) ions. AB - The semicarbazone of 8-hydroxyquinoline-2-carbaldehyde can be easily synthesized and is an effective tetradentate ligand for the coordination of rare-earth(III) ions. Investigations with yttrium(III) and lanthanum(III) in solution and in the solid state show, that the small yttrium ion can form 2 : 2 (1 : 1 stoichiometry) and 2 : 1 ligand to metal complexes (X-ray structures: [LY(NO3)(DMF)2]2Cl2 x 2DMF and [LL'Y] x 3MeOH x Et2O). With the larger lanthanum(III) ion only a well defined 1 : 1 complex (X-ray structure: [LLa(NO3)(MeOH)2]2(NO3)2) can be observed but probably 2 : 1 complexes are also formed. The X-ray structure analyses of [(L H)MCl3] x MeOH (M = Er, Ho) and Na[(micro-NO3){LEu(NO3)2}2] x 2DMF show different coordination modes of the ligand. It can coordinate in its deprotonated but also in the protonated form. PMID- 16471058 TI - Ag(I) complexes with alkylidene-bis(2-aminopyrimidines) as building units for discrete metallomacrocyclic frames. A structural and solution study. AB - Alkylidene-bis(2-aminopyrimidines) (pyr2Cx, x = 2-5) are useful ligands to interact with Ag(I) yielding discrete metallocycles. Crystal structures of the [(pyr2C2)Ag(NO3)]2 and [(H-pyr2C4)Ag(NO3)2]2 have been isolated where each macrocyclic moiety interacts with their surroundings through weak interactions, yielding 3D discrete structures, On the other hand, the solution study shows that the equilibrium constants for the formation of Ag(pyr2Cx)+ complexes are higher than the literature values for Ag(I) complexes with single pyrimidines, although the differences could be explained by invoking the solid-state structures of the Ag(I)-pyr2Cx complexes. PMID- 16471059 TI - Parahydrogen derived illumination of pyridine based coordination products obtained from reactions involving rhodium phosphine complexes. AB - The reactions of RhCl(PBz3)3 with H2 and pyridine or 4-methylpyridine yield RhCl(H)2(PBz3)2(py) and RhCl(H)2(PBz3)2(4-Me-py), respectively. These species undergo hydride site exchange via the loss of the pyridyl donor and formation of RhCl(H)2(PBz3)2 which contains equivalent hydride ligands; for the py system the activation free energy, deltaG++300, is 57.4 +/- 0.1 kJ mol(-1) while for 4-Me-py the value is 59.6 +/- 0.3 kJ mol(-1). These products only showed parahydrogen enhancement in the corresponding hydride resonances when a sacrificial substrate was added to promote hydrogen cycling. When RhCl(PPh3)3 was used as the precursor similar observations were made, while when RhCl(PCy3)2(C2H4) was examined, H2 addition led to the formation of the binuclear complex (H)2Rh(PCy3)2(micro Cl)2Rh(H)2(PCy3)2 which was differentiated from RhCl(H)2(PCy3)2 on the basis of the similarity in diffusion coefficient (5.5 x 10(-9) m2 s(-1)) to that of (H)2Rh(PPh3)2(micro-Cl)2Rh(PPh3)2 (5.3 x 10(-9) m2 s(-1)). The detection of RhCl(H)2(PCy3)2(py) was facilitated when pyridine was added to a solution of RhCl(PCy3)2(C2H4) before the introduction of H2. During these reactions trace amounts of the double substitution products, RhCl(H)2(phosphine)(py)2, were also detected. PMID- 16471060 TI - sigma-->sigma* Transitions in Mn2(CO)8L2 complexes (L = P-donor ligands): pi acidity and oxygen pendant group effects in phosphite ligands. AB - Comparison of the energies assigned to the sigma-->sigma* transitions in bis axial Mn2(CO)8L2 complexes when L = phosphites and phosphinites with those when L = alkyl and aryl phosphines shows that the oxygen-containing ligands do show effects associated with their pi-acidity. However, the data also strongly suggest that the energies are affected by the presence of the oxygen atoms themselves to an extent that increases linearly with the number of oxygen atoms connected to the P-donor atoms. This "oxygen effect" is analogous to, but distinct from, the aryl effect that is also simply proportional to the number of aryl groups attached to the P-donor atoms. This resolves the incongruity of the claim that phosphites also show an "aryl effect", and suggests that specific "pendant group effects" due to the nature and number of atoms attached to the phosphorus atoms of P-donor ligands (aryl, oxygen, nitrogen, sulfur etc.) may be quite general. PMID- 16471061 TI - Ferromagnetism in dinuclear copper(II)-phenolate complexes with exogenous O-donor bridges: a comparative study. AB - The copper(II) complex Cu2L(OAc)(H2O)3.5, 1 x 3.5H2O was obtained and its reactivity in a basic medium investigated. Complex 1 x 3.5H2O shows different reaction patterns in air and in an inert atmosphere. Accordingly, interaction of 1 x 3.5H2O with Me4NOH x 5H2O in methanol-acetonitrile in air yields the hydroxide complex Cu2L(OH)(H2O)1.125, 2 x 1.125H2O while Cu2L(OMe)(MeOH)0.5(H2O), 3 x 0.5MeOH x H2O is isolated under an argon atmosphere. The products 1-3 were fully characterised and single crystals of {[Cu2L(OAc)] x MeCN x 3.5H2O}2, 1 x MeCN x 3.5H2O, {[Cu2L(OH)] x MeCN x 1.125H2O}2, 2 x MeCN x 1.125H2O and [Cu2L(OMe)] x 0.5MeOH x H2O, 3 x 0.5MeOH x H2O solved. The single X-ray study shows that 1-3 are dinuclear complexes with an endogenous phenol oxygen and an exogenous O-bridge. Magnetic characterisation of the three dinuclear complexes was performed, showing an apparent anomalous intramolecular ferromagnetic coupling between the metal atoms in all cases. PMID- 16471062 TI - Electron delocalisation in a trinuclear copper(II) complex: high-field EPR characterization and magnetic properties of Na3[Cu3(mal)3(H2O)] x 8H2O. AB - The complex Na3[Cu3(mal)3(H2O)] x 8H2O was obtained from evaporation of an aqueous solution containing Cu(OAc)2, malic acid (HO2CCH2CHOHCO2H) and NaOH and was characterised by X-ray diffraction on single crystal, X-band and high-field EPR spectroscopy (HF-EPR) and magnetic susceptibility measurements. The trinuclear complex [Cu3(mal)3(H2O)]3- is trapped in a three-dimensional network with sodium cations. The three copper atoms are connected by alkoxo bridges and form an almost isosceles triangle with Cu...Cu distances of 3.076(1), 3.504(1) and 3.513(1) A. Two of the copper ions are also bridged by an extra aquo ligand. EPR spectroscopy combined with magnetic susceptibility measurements provide a powerful tool to resolve the electronic structure of the complex. The overall magnetic behaviour corresponds to an antiferromagnetically coupled triangular system. The 285 GHz-EPR spectrum (g = 2; 10.18 T) is characteristic of a spin state S = 1/2, with a rhombic anisotropy of [g]. This rhombic pattern allows us to propose that the electronic spin density is delocalised on the three copper ions. PMID- 16471063 TI - Synthesis and structural characterization of new oxorhenium and oxotechnetium complexes with XN2S-tetradentate semi-rigid ligands (X = O, S, N). AB - Twelve novel oxo-technetium and oxo-rhenium complexes based on N2S2-, N2SO- or N3S-tetradentate semi-rigid ligands have been synthesised and studied herein. By reacting the ligands with a slight excess of suitable [MO]3+ precursor (ReOCl3(PPh3)2 or [NBu4][99gTcOCl4]), the monoanionic complexes of general formula [MO(Ph-XN2S)]- could be easily produced in high yield. The complexes have been characterized by means of IR, electrospray mass spectrometry, elemental analysis, NMR and conductimetry. The crystal structures of [PPh4][ReO(Ph-ON2S)] 1b and [NBu4][99gTcO(Ph-ON2S)] 1c have been established. The [MO]3+ moiety was coordinated via the two deprotonated amide nitrogens, the oxygen and the terminal sulfur atoms in 1b and 1c. In both compounds, the ON2S coordination set is in the equatorial plane, and the complexes adopted a distorted square-pyramidal geometry with an axial oxo-group. The chemical and structural identity of the different prototypic complexes (rhenium, 99gTc complexes and their corresponding 99mTc radiocomplexes) have been also established by a comparative HPLC study. PMID- 16471064 TI - Unusual binding ability of vancomycin towards Cu2+ ions. AB - Vancomycin, a "last chance" antibiotic, is a glycopeptide consisting of an oligopeptide unit being potentially the effective binder of Cu2+ ions. The potentiometric and spectroscopic studies (UV-Vis, CD, EPR, NMR) have shown that, indeed, the peptide unit binds cupric ions very effectively forming almost instantly the 3N complex involving the N-terminal nitrogen donors in the metal ion coordination. The comparison of the binding ability of vancomycin with other peptide chelators clearly shows the efficiency of this antibiotic in metal ion coordination. It is very likely that Cu2+ ions may play a crucial role in the pharmacology of vancomycin, particularly when administered in high doses. PMID- 16471065 TI - Structural investigation of the bridged activated complex in the reaction between hexachloroiridate(IV) and pentacyanocobaltate(II). AB - Time resolved energy dispersive X-ray absorption spectroscopy has been used to follow the structural evolution of the inner-sphere electron transfer reaction between [IrCl6]2- and [Co(CN)5]3-, and to characterise the local structure of the iridium metal centre in the bridged activated complex formed during the reaction. PMID- 16471066 TI - Pediatric multiple sclerosis. PMID- 16471067 TI - The long-term outlook for essential thrombocythemia. PMID- 16471068 TI - Essential thrombocythemia beyond the first decade: life expectancy, long-term complication rates, and prognostic factors. AB - OBJECTIVE: To describe the long-term natural history of essential thrombocythemia (ET) in terms of life expectancy, risk of disease transformation Into a more aggressive myeloid disorder, and prognostic factors for both survival and disease complications. PATIENTS AND METHODS: The study population consisted of a consecutive cohort of patients seen at the Mayo Clinic In Rochester, Minn, in whom a diagnosis of ET was established before 1992, thus allowing a minimum of 10 years of potential follow-up. The conventional criteria-based diagnosis was confirmed by bone marrow biopsy in all Instances. RESULTS: A total of 322 patients were studied (median age, 54 years; median follow-up, 13.6 years). With a median survival time of 18.9 years, survival in the first decade of disease was similar to that of the control population (risk ratio, 0.72; 95% confidence interval, 0.50-0.99) but became significantly worse thereafter (risk ratio, 2.21; 95% confidence Interval, 1.74-2.76). Multivariable analysis identified age at diagnosis of 60 years or older, leukocytosis, tobacco use, and diabetes mellitus as Independent predictors of poor survival. A 2-variable model based on an age cutoff of 60 years and leukocyte count of 15 x 10(9)/L resulted in 3 risk groups with significant difference in survival. In addition, age at diagnosis of 60 years or older, leukocytosis, and history of thrombosis were independent predictors of major thrombotic events. The risk of leukemic or any myeloid disease transformation was low in the first 10 years (1.4% and 9.1%, respectively) but increased substantially in the second (8.1% and 28.3%, respectively) and third (24.0% and 58.5%, respectively) decades of the disease. CONCLUSION: Life expectancy in patients with ET is significantly worse than that of the control population. Leukocytosis is identified as a novel independent risk factor for both inferior survival and thrombotic events. PMID- 16471069 TI - Silent pain sufferers. AB - OBJECTIVE: To evaluate the proportion and characteristics of patients with chronic pain who do not seek treatment and assess whether these patients have unmet pain care needs. PATIENTS AND METHODS: We performed a cross-sectional survey of residents of Olmsted County, Minnesota, from March through June 2004, with additional visit and diagnosis data from the Rochester Epidemiology Project database. Study participants were a random, population-based sample of eligible adult (>30 years) residents of Olmsted County with at least 1 visit to a local health care facility in the past 3 years. RESULTS: Of the 5897 eligible participants, 3575 people (60.6%) responded. Of the respondents who reported pain of more than 3 months' duration, 497 (22.4%) of the 2211 patients stated that they had not informed their physician about their pain. Of these silent sufferers, 70.6% (351/497) reported having moderate or severe pain, 49.2% (243/497) reported having frequent pain (>8 days per month), and 40.6% (202/497) met both criteria. Silent sufferers also reported that pain interfered with their general activity and sleep to a level only slightly less than the chronic pain sufferers who reported discussing their pain with a physician. Silent sufferers made an average of 5.2 ambulatory physician visits per year, which was less than those who sought physician help for their pain (8.6 ambulatory visits per year; P < .001). Men and younger participants were more likely to be silent about their pain (P < .001). CONCLUSION: More than 1 in 5 people with chronic pain did not seek physician care for their pain. This group is unknown to physicians and therefore represents an unreported patient group with an unmet need for pain care. PMID- 16471070 TI - Diffuse bronchiolar disease due to chronic occult aspiration. AB - OBJECTIVE: To describe clinical, radiological, and histopathologic features of diffuse bronchiolar disease due to chronic occult aspiration. PATIENTS AND METHODS: We identified 4 patients encountered from July 2001 to January 2004 who had persistent respiratory symptoms and lung Infiltrates and who were eventually diagnosed by surgical lung biopsy as having diffuse bronchiolar disease due to chronic occult aspiration. Medical records, radiological studies, and histopathologic specimens were reviewed to assess their clinicoradiologic presentation and diagnostic features. RESULTS: The mean age of these 4 patients was 50 years (age range, 41-59 years), and 2 were women. All presented with persistent dyspnea, cough, and lung infiltrates. Three had a history of gastroesophageal reflux, but only 1 had active symptoms. Chest radiography showed interstitial infiltrates, whereas the predominant finding on computed tomography was numerous centrilobular nodules in all patients. Bronchoscopic lung biopsies had been performed in all patients, and the results were nondiagnostic. Surgical lung biopsy specimens revealed diagnostic features that consisted of bronchiolocentric organizing pneumonia with giant cells that contained material consistent with food in all 4 patients. CONCLUSION: Diffuse bronchiolar disease likely represents an underrecognized form of aspiration-related lung disease and may occur in relatively young Individuals without symptoms suggestive of recurrent aspiration. Radiological features associated with this disorder are distinctively different from those seen in aspiration pneumonia. PMID- 16471071 TI - Association of plasma homocysteine with coronary artery calcification in different categories of coronary heart disease risk. AB - OBJECTIVE: To Investigate the association of plasma homocystelne with coronary artery calcification (CAC) in strata based on 10-year risk of coronary heart disease (CHD) in a cohort enriched in persons with hypertension. PARTICIPANTS AND METHODS: Fasting plasma homocystelne was measured by liquid chromatography electrospray tandem mass spectrometry. Coronary artery calcification was measured noninvasively by electron beam computed tomography and CAC score calculated using the method of Agatston et al. The 10-year CHD risk was calculated based on the Framingham risk score. The association of homocysteine with log-transformed CAC score was assessed in the pooled sample and within each risk stratum by linear regression after adjustment for conventional risk factors. RESULTS: In the 1071 participants studied, homocysteine was associated with CAC quantity (P = .01) after adjustment for CHD risk factors (age, male sex, total and high-density lipoproteln cholesterol, diabetes, history of smoking, body mass Index, and systolic blood pressure), serum creatinine, and statin and hypertension medication use. When the association was assessed in strata based on 10-year CHD risk, homocysteine was significantly (P = .003) associated with CAC quantity in participants at Intermediate 10-year risk of CHD (6%-20%) independent of other risk factors but not in those at lower risk or higher risk. CONCLUSION: Plasma homocysteine is associated with quantity of CAC Independent of CHD risk factors. When studied in categories of 10-year CHD risk, the association was significant in participants at intermediate risk but not in those at low or high risk. Plasma homocysteine levels may have clinical utility as a marker of CHD risk in such individuals. PMID- 16471073 TI - Clinical importance of cardiac troponin release and cardiac abnormalities in patients with supratentorial cerebral hemorrhages. AB - OBJECTIVE: To determine the Incidence of cardiac troponin T (cTnT) elevation, electrocardiographic (ECG) changes, and arrhythmias in supratentorial intracerebral hemorrhage (ICH) and their association with early mortality. PATIENTS AND METHODS: Patients with supratentorial ICHs admitted to Mayo Clinic, Rochester, Minn, from March 1998 to October 2003 were studied. We excluded moribund patients with ICHs who died within 12 hours of hospital admission. Cardiac troponin T levels measured on admission and day 2 were determined by a third-generation enzyme-linked immunosorbent assay. Continuous ECG monitoring was performed In all patients. Computed tomographic scans were graded and correlated with abnormal cardiac variables. RESULTS: Peak levels of cTnT were elevated at 0.035 to 1.2 microg/L (mean +/- SD, 0.27 +/- 0.38 microg/L) in 10 (20%) of 49 patients and were not associated with changes in creatine kinase MB fraction or ECG results. The cTnT levels did not correlate with location or side of hemorrhage or mortality at 30 days. Seventy (64%) of 110 patients displayed ECG abnormalities. The ECG changes did not correlate with the location or side of ICH, hydrocephalus, midline shift, or extension to the ventricles. CONCLUSION: The cTnT elevations in survivors of acute ICH are frequent but without confirmatory ECG changes that suggest mild myocardial injury. One-month mortality is not influenced by such cTnT elevations. In addition, ECG abnormalities are common but likely benign in patients with supratentorial ICH who survive the initial insult. PMID- 16471072 TI - Effect of shifting costs to patients on specialty evaluation for sleep disorders. AB - OBJECTIVE: To determine whether the introduction of out-of-pocket expenses to medical center employees would lead to decreased use of sleep disorder services. PATIENTS AND METHODS: We retrospectively analyzed and compared the clinical and medical accounting data from visits by Mayo Clinic employees to the Sleep Disorders Center from January 1 to March 31, 2003, with that of January 1 to March 31, 2004, le, before and after a January 2004 increase in co-payments for evaluation and testing. RESULTS: The total number of new patients evaluated in the first quarters of 2003 and 2004 was similar (113 vs 119; P = .37). Snoring, restless legs symptoms, hypertension, atrial fibrillation, and prior overnight oximetry testing were more prevalent in 2004 than in 2003 (P = .05, P = .01, P < .001, P = .003, P = .02, respectively). In contrast, insomnia and parasomnia complaints were less common in 2004 (P < .001). The mean apnea-hypopnea index, minimum oxygen saturation, and percentage of time with oxygen saturation less than 90% were all more severe in 2004 (P = .01, P = .001, P < .001, respectively). Sleep-related breathing disorders were more commonly diagnosed in 2004 (83.2% vs 67.2%; P = .02), whereas the diagnoses of nonbreathing disorders declined. CONCLUSION: The insurance policy changes that resulted in larger employee co-payments shifted the spectrum of diagnoses seen at the Sleep Disorders Center toward more symptomatic patients, with more associated comorbidities, and patients who had more severe sleep-related breathing disorders. Total utilization did not decrease. PMID- 16471074 TI - Iron chelation therapy for myelodysplastic syndrome: if and when. PMID- 16471075 TI - National Institutes of Health-sponsored workshop on inflammation and immunity in dilated cardiomyopathy. AB - Nonischemic dilated cardiomyopathy (DCM) is an uncommon cause of heart failure but has widespread importance because it is the cause of 45% of heart transplantations. Multiple experimental and clinical lines of evidence have implicated altered immunity in the pathogenesis of DCM. However, advances in the understanding of the mechanisms of altered immunity have not affected the diagnosis or treatment of DCM. In recognition of this problem, the National Institutes of Health sponsored an expert workshop with 2 aims: to review the current understanding of inflammation and immunity as they relate to DCM and to identify the most promising areas for future clinical research efforts in the field. This report summarizes the scientific opportunities, perceived needs and barriers, and workshop recommendations on research directions in DCM. The major recommendations from the members of the workshop are organized according to the following themes: cardiotropic viruses, innate and acquired immune responses, environmental factors, novel diagnostics, and novel therapeutics. PMID- 16471076 TI - Pharmacological therapy for acute respiratory distress syndrome. AB - Acute respiratory distress syndrome (ARDS) is an Inflammatory process caused by a variety of direct and indirect injuries to the lungs. Despite improvements in supportive care and advances in ventilator management, mortality in patients with ARDS remains high. Multiple pharmacological interventions have been investigated but have not shown improved survival. Clinical trials using corticosterolds, prostaglandins, nitric oxide, prostacyclin, surfactant, lisofylline, ketoconazole, N-acetylcystelne, and fish oil have been unable to show a statistically significant Improvement in patient mortality. As more is understood about the pathophyslology of ARDS, treatment strategies such as increasing alveolar fluid clearance through activation of sodium channels, enhancing repair of alveolar epithelium with growth factors, inhibiting fibrin deposition, blocking proinflammatory transcription factors, preventing the effect of potent vasocontrictors such as endothelin, and using antibodies against key inflammatory cytokines are being explored. This review focuses on the pharmacological treatments studied clinically, proposed reasons for their lack of success, and new concepts emerging in ARDS therapy. PMID- 16471077 TI - Human immunodeficiency virus-associated peripheral neuropathies. AB - Peripheral neuropathy has emerged as the most common neurologic complication of human immunodeficiency virus (HIV) infection. It will continue to play an Important role in HIV Infection given the fact that HIV-infected Individuals are living longer, are at risk of long-term metabolic complications, and face an Increasing exposure to potentially neurotoxic antiretroviral drugs. We review the various types of peripheral neuropathy that have been associated with HIV infection, including distal symmetrical polyneuropathy, toxic neuropathy from antiretroviral drugs, diffuse infiltrative lymphocytosis syndrome, inflammatory demyelinating polyneuropathies, multifocal mononeuropathies, and progressive polyradiculopathy. PMID- 16471078 TI - Dementia and cerebrovascular disease. AB - Cerebrovascular disease is an important cause of cognitive Impairment and dementia in elderly patients. This review highlights the challenges involved in examining the role of cerebrovascular disease In dementia, areas in which consensus is emerging, and an operational framework for clinicians. Two important challenges exist. First, there is no accepted neuropathologic scheme for quantitating cerebrovascular disease In cognitive disturbances. Second, agreement on clinical definitions of vascular dementia is incomplete. Despite the barriers posed by these 2 deficiencies, many consistencies in the clinical, imaging, epidemiological, and neuropathologic aspects of cerebrovascular disease and cognitive impairment have been Identified. Different levels of cerebrovascular disease related to cognitive impairment can be suggested on clinical and Imaging grounds. The overlap between cerebrovascular disease and Alzheimer disease produces a disorder that might be amenable to therapeutic approaches based on either mechanism. PMID- 16471079 TI - Hearing loss and hearing aid treatment options. AB - Hearing loss is a common problem that impacts quality of life for the patient, family members, and caretakers. Unfortunately, hearing loss is often undiagnosed and untreated. This article summarizes symptoms of typical hearing loss associated with aging and simple screening procedures that can be Implemented into a primary care practice. In addition, hearing aid treatment options, Including monaural or binaural fitting, size and style, and circuitry and feature options, are reviewed. PMID- 16471080 TI - Acromegaly with normal insulin-like growth factor I levels. PMID- 16471081 TI - Luis Walter Alvarez: another "Mayo-trained" Nobel Laureate. PMID- 16471082 TI - Adult acute myeloid leukemia. AB - Acute myeloid leukemia (AML) is a group of several different diseases, the treatment and outcome of which depend on several factors, including leukemia karyotype, patient age, and comorbid conditions. Despite advances in understanding the molecular biology of AML, its treatment remains challenging. Standard regimens using cytarabine and anthracyclines for induction followed by some form of postremission therapy produce response rates of 60% to 70%, with less than 20% of all patients achieving long-term disease-free survival. New therapies are emerging based on the definition of specific cytogenetic-molecular abnormalities. Such targeted therapies offer the promise of better antileukemic activity in adult AML. PMID- 16471083 TI - Who chooses the appropriate treatment for hyperhidrosis--physician and patient, or insurer? PMID- 16471084 TI - High-normal platelet factor 4-heparin antibodies and mortality in hemodialysis patients. PMID- 16471085 TI - Clarity of language. PMID- 16471086 TI - Clarity of language. PMID- 16471087 TI - Magnetic insoles. PMID- 16471088 TI - Magnetic insoles. PMID- 16471089 TI - Finasteride: middle-age cure-all for alopecia and benign prostatic hyperplasia? PMID- 16471090 TI - Weekly versus three-weekly cisplatin as an adjunct to radiation therapy in high risk stage I-IIA cervical cancer after surgery: a randomized comparison of treatment compliance. AB - OBJECTIVES: To compare weekly and three-weekly cisplatin as an adjunct to radiation therapy in high-risk early-stage cervical cancer after surgery with regard to treatment compliance. MATERIAL AND METHOD: From June 1st, 2003 to February 29th, 2004, the authors performed a randomized trial of radiotherapy in combination with two concurrent chemotherapy regimens - weekly or three-weekly cisplatin--in patients with high-risk cervical cancer FIGO stage I-IIA after surgery. Women with primary invasive squamous-cell carcinoma, adenocarcinoma, or adenosquamous carcinoma of the cervix were enrolled. The patients also had to have an absolute neutrophil count of at least 1,500 cells per cubic millimeter, a platelet count of at least 75,000 cells per cubic millimeter, a creatinine clearance higher than 40 milliliter per minute, and adequate hepatic function. All patients received external-beam radiotherapy according to a strict protocol. Patients were randomly assigned to receive one of two chemotherapy regimens: 75 mg per square meter of cisplatin on days 1, 22, 43 and 64 or every three weeks for 4 cycles (group 1) or 40 mg per square meter of cisplatin per week for six cycles (group 2). RESULTS: The analysis included 40 women. The first group that received three-weekly cisplatin had a higher rate of incomplete and delayed treatments than the second group that received weekly cisplatin (p < 0.001 and p = 0.0236 respectively). The relative risks of delayed courses were 2.06 (95 percent confidence interval, 1.15 to 3.68) for group 1, compared with group 2. The toxicity-related incomplete treatments rate and G-CSF doses used were significantly higher in group 1 than in group 2. CONCLUSION: Concurrent chemoradiation with weekly cisplatin regimen has more complete treatment rate and less delayed courses than that with three- weekly cisplatin among women with high risk cervical cancer after surgery. PMID- 16471091 TI - Can pre-operative urodynamic study predict the successful outcome of tension free vaginal tape (TVT) operation in Thai women with stress urinary incontinence? AB - OBJECTIVE: To study the preoperative urodryramic parameters whether it can predict the outcome of TVT operation in Thai women with stress urinary incontinence (SUI). STUDY DESIGN: Cohort study. MATERIAL AND METHOD: Twenty five Thai women undergone TVT from January to December 2004 at Bumrungrad Hospital were included in the present study. Urodyramic studies were done pre-operatively. The TVT operations were carried out with spinal anesthesia. The follow-ups at one, three, and six months were scheduled. RESULTS: At six months after the operation, the subjective cure rate was 72% (18/25) and 28% (7/25) had the improved symptoms. Multivariate regression analysis showed the valsava leak point pressure (VLPP) of less than 60 cm H2O to be associated with the 0.6-fold (0.3 0.9) risk of having successful outcome (being cured). Anyhow 7 cases of women not being cured still had improved symptoms and were satisfied with the operation. CONCLUSION: The present results confirmed the usefulness of pre-operative urodynamic study in prediction of the outcome of TVT. Low VLPP (<60 cm. H2O) was associated with poorer outcome of TVT operation in Thai women. PMID- 16471092 TI - Urinary symptoms and quality of life changes in Thai women with overactive bladder after tolterodine treatment. AB - OBJECTIVES: To study the urinary symptoms and quality of life changes in Thai women with overactive bladder (OAB) after tolterodine treatment. MATERIAL AND METHOD: Thirty women (aged 30-77 years) diagnosed as having OAB at the Gynecology Clinic, King Chulalongkorn Memorial Hospital from January to April 2004 were included in the present study. Tolterodine 2 mg, twice daily was given. After 8 weeks treatment, changes in micturition diary variables and tolerability were determined. Short form 36 (SF36) questionaires (Thai version) were given before and after 8 weeks of treatment. RESULTS: At 8 weeks, all micturition per day decreased from 16. 7 +/- 5. 3 to 6. 7 +/- 2.4 times per day. The number of nocturia episodes decreased from 5.4 +/- 4.2 to 1.1 +/- 1.0 times per night. The most common side effect was dry month in 5 cases (16.7%) with 2 cases reporting a moderate degree and 1 case with severe degree. Only one case (3.3%) withdrew from the present study due to a severe dry mouth. The SF-36 scores changed significantly in the domains of physical functioning, role function emotional, social function and mental heath. CONCLUSION: Tolterodine was well tolerated and its effects improved the quality of life in Thai women with OAB. PMID- 16471093 TI - Risk factors of venous thromboembolism (VTE) in Thai reproductive aged female: King Chulalongkorn Memorial Hospital experience. AB - OBJECTIVE: To estimate the risk factors of VTE focusing on oral pill use, alcohol intake and smoking among women who attended at King Chulalongkorn Memorial Hospital between 1 January 1995 and 31 December 2004. MATERIAL AND METHOD: The study design was case-control study. A Case was defined as the female inpatient who was diagnosed with VTE and a control was defined as the healthy female patient who attended gynecological clinic for annual check up. The questionnaires were developed to interview both cases and controls. The ratio of case per control was 1:2. The data was analyzed with SPSS/PC+ version 12.0. The statistic uses were mean, standard deviation, t-test, Odds ratio and 95% confidence interval. RESULTS: Seventy of cases and one hundred and forty of controls were recruited into the study. The mean +/- SD of age of case was 37.2 +/- 9.3 years and the control was 35.6 +/- 9.9 years. There were no significant difference in term of age, parity, weight, height and BMI between cases and controls. The Odds ratio of oral contraceptive pill use, smoking and alcohol consumption were 0.94, 4.28, and 2.62, respectively. However, no statistical significance difference of Odds ratio in oral contraceptive pill use, and alcohol consumption were demonstrated. CONCLUSION: Smoking and alcohol consumption increased risk of VTE in this study. Oral contraceptive pill use did not demonstrate increasing risk. However, there was no statistical significance of odds ratio in alcohol consumption. This result may be due to the small sample of cases. The further study should be recruited with more cases in order to demonstrated the risk factors of these female patients. PMID- 16471094 TI - Perinatal effects of amphetamine and heroin use during pregnancy on the mother and infant. AB - OBJECTIVES: To determine the perinatal impacts of heroin and amphetamine on both mothers and infants. MATERIAL AND METHOD: This is a retrospective study on the influence of amphetamine and heroin on pregnant women and their newborn infants at King Chulalongkorn Memorial Hospital in Bangkok, Thailand, between January 1997 and December 2002. The medical and demographic data of both mothers and infants were evaluated. Comparison of the consistent drug effects of these 2 drugs on the mothers and infants were also performed RESULTS: Two hundred and eleven women were identified There were 178 (84.4%) and 33 (15.6%) women addicted to amphetamine and heroin respectively. Sixty one (28.9%) of them were polydrug users. There were more polydrug users among heroin addicts than amphetamine addicts, (43.7% vs 27.2%, p < 0.05). Poor obstetric history were noted in both groups of women including lack of prenatal care (74.9%), a high incidence of previous abortion (22.3%), positive HIV serology test (11.1%), pre-eclampsia (5.2%), infection (3.3%) and antepartum hemorrhage (1.9%). Drug intoxication was found in 11 amphetamine addicted mothers, whereas 2 heroin addicts developed withdrawal symptoms during intrapartum and postpartum periods. All infants were singleton. There was one stillbirth and 2 neonatal deaths. There was no statistical difference in terms of sex ratio, mean birth weight, gestational age, length, head circumference and Apgar score between the groups of amphetamine and heroin exposed infants. The incidence of prematurity, low birth weight, IUGR and microcephaly were not statistically different between both groups of infants. The overall incidence was 31.7%, 31.7%, 9.5% and 8.6% respectively. Congenital anomalies were found in 5 (2.8%) amphetamine exposed infants. Thirty one out of 33 heroin exposed infants (93.9%) and 4 out of 178 amphetamine exposed infants (2.2%) developed drug withdrawal symptoms with the mean onset of 21.5 +/- 16.5 hours and 10.3 +/- 7.5 hours respectively, p > 0.05. All heroin withdrawal infants were successfully treated with Phenobarbital with the mean duration of treatment of 23.7 +/- 11.5 days. None of the amphetamine withdrawal infants needed specific treatment. They recovered spontaneously within 6.0 +/- 5.3 days. Eighteen infants were left in an orphanage or under the custody of their relatives. CONCLUSION: Amphetamine or heroin use during pregnancy can cause many serious adverse effects on both mothers and infants. The findings in the present study are consistent with previous reports, although they seemed to be more common and severe. Increasing awareness and improving understanding of drug abuse in the medical, legal and social aspects are needed in order to reduce these impacts. PMID- 16471095 TI - Risk factor of birth weight below 2,500 grams and attention deficit hyperactivity disorder in Thai children. AB - OBJECTIVES: To study the factor of birth weight below 2,500 grams and Attention Deficit Hyperactivity Disorder (ADHD) and to identify the factors that are associated with ADHD. MATERIAL AND METHOD: A retrospective, hospital-based, case control study was conducted with 122 ADHD cases from the child and adolescent psychiatric outpatient unit at Siriraj Hospital, 119 nonADHD from students who have been assessedfrom Child Behavior Checklist (CBCL) and Conners Teacher Rating Scale, they are identified as normal. Mothers of both groups have been asked to fill a self report questionnaire to evaluate demographic data, prenatal data, factors effected pregnancy both physical and mental illness, post natal period, birth weight, underlying disease, physical illness and family history of ADHD. Multiple logistic regression analysis was used to determine the adjust effect of all factors. RESULTS: The number of ADHD cases who had a birth weight below 2,500 grams was 3.6 times the number of control cases who had a birth weight below 2,500 grams. It was statistically significant (p = 0.03). Factors associated with ADHD were pregnancy complication (p value < 0.05, OR = 4.17, 95%CI [1.66, 14.02]), emotional distress during pregnancy (p value <0.05, OR = 2.99, 95%CI [1.43, 5.40]), postnatal complication (p value < 0.05, OR = 3.26, 95%CI[1.56, 6.41] and a family history of ADHD (p value < 0.05, OR = 3.6, 95%CI [1.65, 8.11]. CONCLUSION: From the present study, the birth weight below 2,500 grams, pregnancy complication, emotional distress during pregnancy, neonatal complication and a family history of ADHD may be the risk factors of ADHD. PMID- 16471096 TI - Outcome of maternal syphilis at Rajavithi Hospital on offsprings. AB - BACKGROUND: Syphilis remains an important sexually transmitted disease and continues to be an important problem in Thailand. Despite the clinical efficiency of penicillin in the treatment of pregnant patients with syphilis, infants with congenital syphilis are still encountered. Congenital syphilis poses significant challenges for the clinician because infants may be asymptomatic at birth or present with a highly variable clinical picture. OBJECTIVES: To evaluate the outcomes of neonates born to syphilitic mothers, the efficacy of antepartum treatment in the prevention of congenital syphilis and treatment for congenital syphilis after delivery. MATERIAL AND METHOD: The surveillance conducted from September 1st, 2002 to December 31st, 2003, involved 63 mothers who were diagnosed with syphilis and their offsprings at Rajavithi Hospital, Bangkok, Thailand. Sixty-four infants had complete physical examination, growth, development and laboratory evaluation at Queen Sirikit National Institute of Child Health at the time of delivery and at the ages of 1, 2, 4 and 6 months. RESULTS: There were 63 mothers and 64 infants recruited in the present study. Fifty-three mothers had prenatal care (84.13%). The VDRL was positive in the first prenatal care visit in 42 mothers (66.67%) and 11 mothers (17.46%) had seroconversion later on. Maternal treatment for syphilis included adequate penicillin 23 cases (36.51%), inadequate penicillin 5 cases (7.94%), erythromycin 9 cases (14.29%) and 26 mothers (41.27%) received no treatment at all. The mean maternal age, mean gestation age at treatment for syphilis and at delivery were 30.31 +/- 5.60 years, 32.75 +/- 6. 73 weeks and 38.60 +/- 1.57 weeks respectively. Failure rate in the adequate penicillin group was 8.7%. The mean birth weight of the 64 infants was 3034 +/- 495 grams, no syphilitic stillbirth occurred. Nine infants (14.06%) were identified with presumptive congenital syphilis. The manifestation include hepatomegaly (55.56%), desquamation of palms and soles (44.44%), radiological changes (33.33%) and abnormal cerebrospinal fluid (25%). The fluorescent treponemal antibody absorption immunoglobulin M (FTA ABS IgM) tests of the infants were positive in 2 out of 9 cases (22.22%). The range of maternal and neonatal VDRL titer were between weakly reactive to 1.32 and nonreactive to 1:32 respectively. Fifty infants (78.13%) including 9 presumptive cases were followed-up, all had normal growth. Thirty-four infants (68%) who had re-evaluation for VDRL titers, were seronegative. CONCLUSION: Penicillin is the effective treatment of pregnant patients with syphilis and infants with congenital syphilis. The high risk of congenital syphilis correlates with untreated mothers and inadequate maternal syphilis treatment. PMID- 16471097 TI - Reliability of a Thai version of King's Health Questionnaire in Thai females with overactive bladder symptoms. AB - OBJECTIVES: To develop and test the reliability of a Thai version of the King's Health Questionnaire (KHQ). MATERIAL AND METHOD: Three Thai Urologists forward translated the original English KHQ into a Thai version. Back translations were performed by an independent group of physicians. A consensus was reached on a final Thai version after comparing the original KHQ and various translations. Fifty Thai female patients with symptoms of overactive bladder were tested and retested every two weeks using the Thai version (twice) as well as the English version (once) of the KHQ. Test-retest reliability of the Thai questionnaire was measured using the kappa statistic. RESULTS AND CONCLUSION: The Thai version of the KHQ was found to be reasonably reliable for use in Thai female patients with over active bladder symptoms. PMID- 16471098 TI - Comparative study between preoperative and postoperative histologic grading in adenocarcinoma of the colon and rectum. AB - BACKGROUND: Preoperative histologic grading was used to decide the length of the margin of resection in adenocarcinoma of colon and rectum. OBJECTIVES: The aim of the present study was to determine the accuracy of preoperative histologic grading by comparison with postoperative histologic grading. MATERIAL AND METHOD: Preoperative and postoperative histologic grading of patients with adenocarcinoma of the colon and rectum in the department of surgery, Phramongkutklao Hospital between January 1999 and October 2004 were collected and compared together using percentage and chi-square test. RESULTS: 260 patients were included in the present study. 47.3% of all cases had inaccurate preoperative histologic grading. 43% of all cases had worse differentiation. 52.7% of all cases had the same differentiation. Only 4.3% of all cases had better differentiation. CONCLUSION: Preoperative histologic grading was not accurate and could not be used in deciding the length of the margin of resection. PMID- 16471100 TI - Positioning of femoral tunnel in anterior cruciate ligament reconstruction using femoral aimer guide. AB - OBJECTIVES: Anterior Cruciate Ligament (ACL) reconstruction is one of the common procedures. A larger number of ACL injury patients seek treatment to return to preinjury level. Many factors affect the result of reconstruction. Femoral position is one of the important factors. Nowadays, Femoral Aimer Guide is used in to find the proper position of the femoral tunnel but Grontvedt, et al reported the technique was unsatisfactory. Some studies show it is impossible to use the device via transtibial technique for anatomical attachment at femoral site. The authors studied the femoral entry point by using the Femoral Aimer Guide both through transtibial tunnel and medial arthrotomy approach. MATERIAL AND METHOD: The authors dissected 37 cadaveric knees, removed the ACL and identified the center of the ACL attachment at the femoral side. Used the Femoral Aimer Guide in assisted to find the position of the femoral entry point at 90 degrees position. Right knee by transtibial technique and left knee by medial arthrotomy technique. Both positions were compared RESULTS: All positions from the Femoral Aimer Guide entry point did not coincide with isometric point. They tended to move more superior and posterior positions. Some of the aimer positions were far more posterior, and caused the posterior cortex to be thinner than 5 mm. CONCLUSION: The Femoral Aimer Guide couldn't find the proper position of graft attachment at the femoral side by the standard technique. PMID- 16471099 TI - Outcome of in-patient trauma cases: Accident and Emergency Unit, Khon Kaen University. AB - OBJECTIVES: To study and report the outcome of in-patient trauma cases based on the Trauma and Injury Severity Scoring (TRISS) method and compare the outcome with the registry data from the Major Trauma Outcome Study (MTOS). MATERIAL AND METHOD: A descriptive study was performed by retrospective data collection. From 1 January 2002 to 31 December 2002, all admitted trauma patients in the Accident and Emergency Unit, Department of Surgery, Faculty of Medicine, Khon Kaen University were included in the present study. Survival analysis was completed for all of the patients. STATISTICAL ANALYSIS: TRISS method and W, M and Z statistics (Z-score) on the basis of definitive outcome-based evaluation (DEF) method for comparing with MTOS data. RESULTS: The majorities of patients were men (76.85%) and mean age was 30.81 years. One hundred and ninety five patients (96.06%) sustained blunt trauma, the vast majority resulting from motor vehicle crashes. The observed survivors were 182, whereas the expected survivors were 183.582. The W, M and Z-statistics were -0.779, 0.843 and -0.493 respectively. CONCLUSION: Z-score -0.493 indicated no statistical difference between observed and expected survivors. PMID- 16471101 TI - Anatomic safe zone of pin insertion point for distal clavicle fixation. AB - Clavicle fracture is the most common childhood fracture and one of the most common fractures in adults. Only some types of distal clavicular fractures, and dislocation of the acromioclavicular joint, require internal fixation. Many surgeons prefer closed pinning; however, the difficulty inserting many of the various kinds of pins from acromion into the medullary canal, of the distal clavicle, means the likelihood of iatrogenic complications from repeated drilling is heightened. The purpose of the present study was to establish what would be the optimum insertion point and direction for safe intramedullary pinning of the distal clavicle. Embalmed cadaveric shoulders (32) were studied. A bone window was created at the distal one-thirds of the clavicle, approximately 1.5 cm medial from the conoid tuberosity - as wide as could be freely, retrogradely drilled into the medullary canal of the distal clavicle. A 2.0-mm Kirschner wire was inserted until it penetrated the acromion. The point of emergence was recorded as ratio compared with the acromial width and length in coronal and sagittal planes, respectively. K-wire directions were measured as the angle between the K-wire and the reference line from the anterosuperior tubercle of the clavicle to the anterior angle of the acromion. The process was repeated until the acromion fractured 304 drillings were performed on 32 specimens. The length of the sagittal vs.coronal pin insertion point from the anterior vs. lateral borders of the acromion divided by its length vs. width averaged 0.325 +/- 0.04 and 0.397 +/ 0.09, respectively. The angle of the K-wire and the reference was 7.69 +/- 3.04 and 14.59 +/- 4.34 degrees in the coronal and horizontal planes, respectively. At 8 and 10 drillings survival was 0.72 (95%CI: 0.53-0.84) and 0.41 (95%CI: 0.24 0.57), respectively. The optimum pin inserting point for fixation of distal clavicle fracture and acromioclavicular joint dislocation is 32.5% and 39.7% of acromial length and width, respectively. If a 2.0-mm K-wire is used for fixation, drilling should not be repeated drilled more than 8 times to avoid sudden, high risk iatrogenic acromial fracture. PMID- 16471102 TI - Parecoxib versus tramadol for post-appendectomy pain. AB - Open uncomplicated appendectomy is known for low to medium degree of postoperative pain and a short hospital stay. Based on multimodal pain therapy, non-opioid analgesics have widely been a part in pain control. Parecoxib and tramadol have advantages over traditional opioids that are causing less nausea or vomiting, respiratory depression and sedation. As a result, the authors aimed to compare parecoxib and tramadol regarding quality of pain control after open appendectomy. Fifty patients, underwent open appendectomy with spinal anesthesia, were randomized to receive either parecoxib or tramadol (n = 25 each). Parecoxib 40 mg and tramadol 50 mg IV were administered twice, when closing the peritoneum and at 12 h later Doses of rescued meperidine for 24 h were recorded. Pain score, sedation, nausea or vomiting and satisfaction scores were assessed at 6, 12 and 24 h after operation. The mean rescued doses of meperidine were 4.6 +/- 10.9 and 18.6 +/- 21.0 mg in parecoxib and tramadol groups respectively (p = 0.005). There was a significantly higher pain score at 24 h (p = 0.01) and sedation score at 6 h (p = 0.003) in the tramadol group. Parecoxib provided a lower pain and sedation scores and lesser meperidine consumption than tramadol for post-appendectomy pain. IMPLICATION: Parecoxib, as a primary analgesic, is better in analgesia and has less sedation than tramadolfor post-appendectomy pain. PMID- 16471103 TI - Levobupivacaine versus racemic bupivacaine for extradural anesthesia for cesarean delivery. AB - BACKGROUND AND OBJECTIVES: Bupivacaine is available as a racemic mixture of dextrobupivacaine and levobupivacaine. Many studies show that dextrobupivacaine has a greater inherent central nervous system and cardiovascular toxicity than levobupivacaine. The aim of the present study was to investigate the clinical efficacy and safety of levobupivacaine compared with racemic bupivacaine for extradural anesthesia. METERIAL AND METHOD: The authors studied 61 patients undergoing elective cesarean delivery who received either 0.5% levobupivacaine (n = 31) or 0.5% bupivacaine (n = 30) extradurally, in a randomized, double blind study. RESULTS: The 2 groups were similar in terms of time to block suitable for surgery, duration of sensory block, time to T10 regression, time to onset and offset of motor block, verbal numeric pain scores at abdominal opening and at child birth. Mean (SD) dose of 0.5% levobupivacaine and 0.5% bupivacaine were 19.3 (4.6) ml and 17.3 (3.8) ml respectively, p = 0.069. CONCLUSION: Levobupivacaine produces an extradural block that is similar to bupivacaine, and is an alternative to bupivacaine for cesarean delivery patients. PMID- 16471104 TI - Comparison of 25 and 27 gauge needle in spinal anesthesia learning curve for anesthesia residency training. AB - BACKGROUND AND RATIONALE: Size of spinal needle may be a factor which influences the success rate of spinal anesthesia. OBJECTIVES: To compare learning curves of using 25G and 27G quincke spinal needle for spinal anesthesia. SETTING: Department of Anesthesiology, Faculty of Medicine, Chulalongkorn University and King Chulalongkorn Memorial Hospital. DESIGN: Prospective randomized control trial. MATERIAL AND METHOD: Ten new first year anesthesia residents were randomized to 2 groups: Group I (n = 5) used 25G Quincke spinal needle, Group II (n = 5) used 27G Quincke spinal needle to perform 200 consecutive cases of spinal anesthesia. Number of success and failure was recorded by each individual resident anonymously. The learning curves of plotted by cumulative sum of success rate revealed an initial rapid improvement of success during the first 20 cases in both groups. The overall success rate of group I was 84% (95CI, 66.5-100) and group II was 87% (95%CI, 61.5-100); p = 0.89. The widest difference between the 2 learning curves at 20 attempts intervals was at the twentieth attempts with a success rate of 76.0% VS 65.0% in group I and group II respectively; p = 0.54. CONCLUSION: The learning curves of spinal anesthesia using 25G and 27G Quincke spinal needle showed rapid improvement of success rate at the first 20 spinal block and high overall success rate with no significant difference between the groups. PMID- 16471105 TI - Cerebral electrical activity as a tool in evaluating anesthetic effect during balanced anesthesia with sevoflurane. AB - OBJECTIVES: The present study was undertaken to provide basic information about bilateral frontal cerebral electrical activity after induction, before and after skin incision, and at a steady state during sevoflurane anesthesia at the end tidal concentration 1, 1.2, 1.4 and 1.6 MAC and determine the association between the electrical cerebral activity with other clinical end points, i.e. motor responses and post-operative recall. MATERIAL AND METHOD: the Dual Channel Brain Activity, ABM2 (DATEX) was used to continuously monitor frontal EMG and electrical cerebral activities (i.e. frequency and amplitude) of both hemispheres in 20 adult female patients undergoing balanced anesthesia in Maharaj Chiang Mai Hospital. The eligible patients were randomly assigned to receive 1.0 MAC, 1.2MAC, 1.4MAC or 1.6MAC of a mixture of sevoflurane and 66.67% nitrous oxide in oxygen during anesthesia. The anesthesia was induced with 5 mg/kg thiopental and supplemented with 1-1.5 microgram per kilogram of fentanyl. The standard dose of pancuronium or atracurium was given during maintenance. RESULTS: After induction with 5 mg/kg thiopental, the mean (95% confidence interval) of frontal EMG significantly decreased from 2.66 (1.63,4.29) to 1.41 (0.2,1.61). When eyelash reflex was absent, the mean (95%CI) frequency and amplitude of the right frontal EEG was 3.89 (3.29, 4.497) Hz, 39.58 (32.11, 47.05) microvolt and left frontal EEG was 3.84 (3.43, 4.25) Hz, 33.55 (28.59, 38.61) microvolt. The findings were consistent with the raw EEG shown on the monitor, i.e. a progressive decrease in the frequency and an increase in the amplitude. During maintenance with the inhaled anesthetics, there was a statistically significant decrease in frequency of right frontal in those who had received the inhaled anesthetic concentration to reach 1.4 and 1.65 MAC (p < 0.05) (repeated measure ANOVA). At the steady state of end tidal concentration of the sevoflurane there was consistency in decreasing frequencies and increasing amplitudes of both hemispheres of the groups with higher MAC values (p < 0.05) (Table 3). However, the authors failed to demonstrate the relationships between EEG changes and other clinical responses. CONCLUSION: The present study has provided basic information about cerebral electrical activity during the balanced anesthesia with sevoflurane. As anesthesia deepened by increased MAC, the frequency decreased and the amplitude increased. PMID- 16471106 TI - Long-term results of early trabeculectomy with mitomycin-C and subsequent posterior segment intervention in the treatment of neovascular glaucoma with hazy ocular media. AB - OBJECTIVES: To assess the long-term efficacy and safety of early trabeculectomy with mitomycin-C (TMMC) and subsequent posterior segment intervention in the treatment of neovascular glaucoma (NVG) with hazy ocular media. MATERIAL AND METHOD: Twenty-three eyes of 21 patients who had NVG with hazy ocular media that precluded posterior segment and had persistent IOP of 30 mm Hg or more despitefully antiglaucoma medications for 48 hours, underwent a modified TMMC (twice application of subconjunctival and subscleral MMC 0.2 mg/ml for 3 and 2 minutes, small internal block excision, laser suturelysis at 2 weeks) and subsequent posterior segment intervention with or without phacoemulsification. RESULTS: Preoperative IOP ranged from 30-80 mmHg, (mean 38.87 +/- 9.52). Follow up period ranged from 12-47 months (mean 29 +/- 11.03). At final follow-up, qualified success (an IOP of < or = 21 mm Hg with or without medication) and complete success (an IOP of < or = 15 mm Hg without medication) was achieved in 21 (91.3%) of 23 eyes and in 12 (52.7%) of 23 eyes, respectively. The median successful period of maintaining qualified success was 13 months. Final visual acuity of 20/400 or better was preserved in 12 of 23 eyes (52.7%). None had hypotony maculopathy, leaking blebs or endophthalmitis. CONCLUSION: Early TMMC and subsequent posterior segment intervention, offerred a safe and prolonged satisfactory result of IOP control in NVG patients with hazy ocular media. PMID- 16471107 TI - A comparison between endoscopically middle meatal aspiration culture using modified aspiration instrument and direct maxillary antral tap culture in chronic rhinosinusitis. AB - OBJECTIVES: To evaluate the accuracy of endoscopically guided middle meatal aspiration culture by comparing the culture results between middle meatal aspiration using the modified aspiration instrument and direct maxillary antral tap. MATERIAL AND METHOD: Sixteen patients with chronic rhinosinusitis underwent functional endoscopic sinus surgery (FESS) were enrolled. Both endoscopically middle meatal aspiration culture (EMAC) using modified aspiration instrument and direct antral tap culture (ATC) were performed before FESS. Microbiologic data were compared and analyzed for any statistical differences between EMAC and ATC. RESULTS: The positive culture rates were 93.75% in both EMAC and ATC groups. Aerobic and facultative anaerobic bacteria were found in 87.5% of EMAC group and 81.25% of ATC group. The two most common bacteria in both groups were coagulase negative Staphylococcus and Staphylococcus aureus. The association between EMAC and ATC was strong to moderate (13/16) 81.25%. CONCLUSION: EMAC appears to be a valuable alternative to ATC for guiding bacterial-specific therapy in chronic rhinosinusitis. This modified aspiration instrument should be useful in clinical practice and serve as a cost effective procedure. PMID- 16471108 TI - Validity and reliability of the Young Mania Rating Scale: Thai version. AB - OBJECTIVES: The objective of the present study was to assess the reliability and validity of the Young Mania Rating Scale (YMRS) Thai Version in 76 patients with known manic symptoms. The study was carried out at Outpatient and Inpatient Departments of Psychiatry, Ramatibodhi Hospital, Bangkok, Thailand MATERIAL AND METHOD: Cross-sectional study. A Thai version of the YMRS was developed through forward-backward translation techniques and reviewed by five content-experts using psychometric methods to test the reliability and validity of the version. RESULTS: An eleven-item questionnaire was developed The validity was established with the item coefficient > 0.6for all scales. The reliability of the YMRS Thai Version on a interrater test had a Pearson's correlation of 0.87 and a Cronbach's Alpha coefficient for all scales of > 0.70. The scale's Cronbach's Alpha coefficient for the total number of items was 0.89. CONCLUSION: The present study supports the use of the YMRS Thai Version as a valid measurement for mania in Thai patients. PMID- 16471110 TI - Health status of health care workers at Srinagarind Hospital: experience from the Annual Health Check-up Program. AB - The authors conducted this study to evaluate the efficacy of a health check-up program and the health status of health care workers at Srinagarind Hospital, Faculty of Medicine, Khon Kaen University. The authors reviewed all yearly-check up charts of personnel who worked at Srinagarind Hospital from 2002 to 2003 including history taking, physical examination, and laboratory testings. There were 606 office workers and 1,024 nursing staff enrolled The mean ages of both groups were 38.9 and 36.5 years old, respectively. The office workers visited physicians significantly more often than the nursing staff (553 of 606 vs 271 of 1,024; p-value = 0.00). Obesity was found much more in office workers (127 of 472 versus 129 of 749). There were significant differences between the groups on impaired fasting plasma glucose, DM, HT, high cholesterol level, high triglyceride level, and significant elevation of ALT or AST (all p-value = 0.00). In the obese group, there was also a significantly higher number of cases who had high blood pressure, defined as IFG or DM, high cholesterol level, and high triglyceride level (p-value = 0.00) except the significant elevation of ALT or AST level. Cases of obesity with significant elevation of hepatic enzyme had many atherosclerotic risk factors. Therefore, metabolic derangements are the important problem for health care workers. PMID- 16471109 TI - The development of the Pictorial Thai Quality of Life. AB - BACKGROUND: "Quality of life" has become a main focus of interest in medicine. The Pictorial Thai Quality of Life (PTQL) was developed in order to measure the Thai mental illness both in a clinical setting and community. OBJECTIVES: The purpose of this study was to develop the Pictorial Thai Quality of Life (PTQL), having adequate and sufficient construct validity, discriminant power, concurrent validity, and reliability. MATERIAL AND METHOD: To develop the Pictorial Thai Quality of Life Test, two samples groups were used in the present study: (1) pilot study samples: 30 samples and (2) survey samples were 672 samples consisting of normal, and psychiatric patients. The developing tests items were collected from a review of the literature in which all the items were based on the WHO definition of Quality of Life. Then, experts judgment by the Delphi technique was used in the first stage. After that a pilot study was used to evaluate the testing administration, and wording of the tests items. The final stage was collected data from the survey samples. RESULTS: The results of the present study showed that the final test was composed 25 items. The construct validity of this test consists of six domains: Physical, Cognitive, Affective, Social Function, Economic and Self-Esteem. All the PTQL items have sufficient discriminant power It was found to be statistically significant different at the. 001 level between those people with mental disorders and normal people. There was a high level of concurrent validity association with WHOQOL-BREF, Pearson correlation coefficient and Area under ROC curve were 0.92 and 0.97 respectively. The reliability coefficients for the Alpha coefficients of the PTQL total test was 0.88. The values of the six scales were from 0.81 to 0:91. CONCLUSIONS: The present study was directed at developing an effective psychometric properties pictorial quality of life questionnaire. The result will be a more direct and meaningful application of an instrument to detect the mental health illness poor quality of life in Thai communities. PMID- 16471111 TI - Microalbuminuria prevalence study (MAPS) in hypertensive patients with type 2 diabetes in Thailand. AB - BACKGROUND: Microalbuminuria represents the earliest clinical evidence of diabetic nephropathy, and is a marker of increased cardiovascular (CV) morbidity and mortality. OBJECTIVES: This analysis of Thai data from the Microalbuminuria Prevalence Study (MAPS) assessed the prevalence of macroalbuminuria and microalbuminuria in hypertensive patients with type 2 diabetes. DESIGN: Cross sectional clinic-based epidemiological study. MATERIAL AND METHOD: A total of 100 patients were enrolled, of which 97 patients constituted the per-protocol population (patients with bacteriuria and haematuria were excluded). Patients attended one study visit with no follow-up. RESULTS: Overall, the prevalence of diabetic kidney disease was high, with macroalbuminuria contributing 13.4% [9.9 16.9; 95% confidence interval (CI)] and microalbuminuria contributing 43.3% [38.3 48.3; 95%CI]. CONCLUSION: Annual screening for microalbuminuria is recommended for all patients with type 2 diabetes, as early treatment is critical for reducing CV risks. Clinical studies have shown that renin-angiotensin system inhibitors can slow the progression of diabetic nephropathy. PMID- 16471112 TI - Usage of and cost of complementary/alternative medicine in diabetic patients. AB - The purposes of the present survey research in diabetic patients were 1) to determine characteristics of complementary/alternative medicine (CAM) use, 2) to identify factors related to CAM use such as sociodemographic, adverse effects, and quality of life, and 3) to determine differences between patients who used and did not use CAM. The data was collected through developed questionnaires and SF-36 scale Thai version. Samples were 159 diabetes patients over 18 years of age or older who came for treatment at Suppasitthiprasong Hospital, Ubon Ratchathani Province, Thailand. The results indicated that the prevalence of CAM use was rather high (47.8%). The most common types of CAM used were yoga/exercise (32.8%), unchanged form of herbal medicine (29.9%), and changed form herbal medicine (17.8%). The average expense of CAM use was dollar 8.58 per person per month. Thus, if the percentage of CAM use and the cost were true for other Thai diabetic patients throughout Thailand, CAM use expenditure for the whole country would be about dollar 915,250-1,545,750 per month, which is quite high for a small country like Thailand. Most patients (64.4%) who used CAM did not inform their doctors about their CAM use. Results also indicated that government official patients were more likely to use CAM than those of farmer patients significantly (p-value = 0.03, odds ratio = 12.11). In addition, the present study found that patients who had a higher income were more likely to use CAM than those of lower income patients significantly (p - value = 0.04, odds ratio = 1.01). However, other factors such as age, sex, marital status, level of education, health insurance coverage status, duration of time to treat, occurrence of adverse effects, and quality of life were not different between the patients who used CAM and who did not use CAM. Physicians should pay more attention to the CAM use of patients since they used CAM without informing physicians and some herbal medicines may cause hypoglycemia. However, the study results had some limitations to apply to other Thai populations since the sample were Suppasitthiprasong patients who may be different from other Thai populations in many ways such as their local culture, belief, and CAM use types and cost. PMID- 16471113 TI - Appropriateness of intravenous loading dose of phenytoin treatment in Srinagarind Hospital. AB - Intravenous loading dose of phenytoin treatment (ILP) is a useful treatment but may cause serious adverse events. The present study assessed the appropriate use of ILP in Srinagarind Hospital. The authors reviewed all charts that ILP was ordered between January 1st, 2000 and December 31st, 2001, about indication, the infusion rate, and side effects. There were 206 cases treated with ILP Thirty-two cases (15.7%) received inappropriate treatment by ILP The most common indication was primary prophylaxis before brain surgery. There were 7 cases that developed side effects with 5 cases of high blood phenytoin level. These data showed that physicians should consider more carefully the use of ILP. PMID- 16471114 TI - Drug eruptions at five institutes in Bangkok. AB - The study was performed in five hospitals in Bangkok for a period of one year. All in- and outpatients who developed drug eruption from January to December 2001 were enrolled into the study. Physical examinations and complete history-taking were performed by one of the authors. A skin biopsy was done to confirm the diagnosis in every suspected case. Oral challenge test was performed to obtain a definite diagnosis only in some patients with informed consent. Among 212 patients, the most common causative drugs were antimicrobial agents with cephalosporin group in the highest rank. Maculopapular rash was the most common type of drug eruption followed by urticaria and photosensitivity reaction. It was concluded that antimicrobial agents were the predominant causative agents and maculopapular eruption was the most frequent clinical manifestation. New kinds of antimicrobial agents, anti-inflammatory drugs and lipid lowering agents could cause various patterns of drug eruption. PMID- 16471115 TI - Chrono impact versus enteric coated valproate in Thai epileptic patients. AB - BACKGROUND: Since its first clinical use more than 30 years ago, Valproic acid is still being widely prescribed It has been available in Thailand for more than 20 years. Sodium valproate slow-released (SVSR) form has been used in clinical practice in Thailand since 1990. The objectives of this open study were to access the compliance and satisfaction consequences in the epileptic patients. MATERIAL AND METHOD: In this prospective, multi-center study, the authors compared the compliance and satisfaction consequences in epileptic patients switched from more than two times daily sodium valproate enteric-coated tablet (SVEC) regimen to the same total daily dose of SVSR form given once or twice daily. RESULTS: Eighty nine of the 100 patients completed the study. 43.8% were male (39 of 89 patients). Mean age was 34.74 +/- 12.67 years. Most common etiology of epilepsy was idiopathic 40.4%. Patients were very/fairly happy with the SVSR form 94.4% compared to the SVEC form 56.2% (p = 0.000). Patients had been experiencing no problem with the SVSR form 67.4% compared to SVEC form 38.2% (p = 0.000) and also never missing taking SVSR. form 77.5% compared to SVEC form 40.4% (p = 0.000). According to convenience, patients preferred to administer SVSR form once a day 92.1% and never over taking dosed the antiepileptic drug 96.6%. SVSR form had fewer side effects than the enteric-coated form interms of memory problem (40.4% vs 48.3%) (p = 0.000), sleepiness (30.3% vs 42.7%) (p = 0.041) and difficulty in thinking clearly (38.2% vs 44.9%) (p = 0.001). The patients were seizure free during the study period comparing SVSR form 76.4% to SVEC form 65.2% (p = 0.011). CONCLUSION: Patients preferred once daily regime. Switching from SVEC to SVSR form increased seizure free, reduced side effects, improved patient's compliance and satisfaction. PMID- 16471116 TI - The possible diagnostic role of endoscopic ultrasound in patients with dyspepsia. AB - BACKGROUND: Dyspepsia is common in clinical practice with frequent relapses and often requires multiple investigations to assess intraluminal and extraluminal etiologies. Endoscopic ultrasound (EUS) has the potential of serving both purposes in a single setting. MATERIAL AND METHOD: Patients with dyspepsia who underwent EUS in four-year period were retrospectively reviewed. Diagnostic findings of US, final diagnoses were noted and compared with the reference standards. RESULTS: 131 patients with a mean age +/- SD of 50 +/- 12.7 years were included. The common diagnoses were non-ulcer dyspepsia in 56%, symptomatic gallstone (GS) in 22.9%. EUS detected two GS missed by transabdominal ultrasound (TUS). EUS missed one GS documented by surgery. EUS detected seven cases of ERCP proven CBD stones undetected by TUS and had a sensitivity, specificity, positive predictive value and negative predictive value for CBD stones of 87.5%, 91.7%, 87.5%, and 91.7% respectively. CONCLUSION: EUS is a potential investigation for the management of dyspepsia. PMID- 16471117 TI - Spinal bone mineral density by quantitative computed tomography in Thais compared with Westerners. AB - The present study was to determine age- and gender-based spinal bone mineral density (BMD) by quantitative computed tomography (QCT) in Thais and to compare it with that of Westerners. Four hundred and twenty five healthy Thais, age 20 to 76 years (322 females, mean age 43.4 years; 103 males, mean age 42.8 years) were recruited for BMD assessment by QCT. Spinal BMD peaks in the 20-29 year age group in both genders with a mean value of 171.9 mg/cu cm in females and 171.0 mg/cu cm in males. It subsequently decreases in older age groups. Males' BMD has a higher rate of decline than females' until age 40-49. It then stabilizes from 50-59 while females' BMD shows the highest rate decline at this period After the mid 50's, both genders have bone loss from aging. Compared to Westerners, peak bone mass is reached in the same age group in both genders. Peak bone mass of Thai females is significantly higher than Western females, but that of Thai males is not significantly higher than Western males. BMD of Thai males in the 50-59 and over-60 age groups is higher than that of Westerners. The findings suggest that the cut-off points for osteopenia and osteoporosis, in Thai females are at spinal BMD lower than 143.6 mg/cu cm and 101.15 mg/cu cm, respectively. While the values lie at 143.2 mg/cu cm and 101.5 mg/cu cm among Thai males for osteopenia and osteoporosis, respectively. PMID- 16471118 TI - Radiographic manifestations of pulmonary cryptococcosis. AB - Pulmonary cryptococcosis may occur in both immunocompromised and immunocompetent hosts. The purpose of the present study was to review the radiologic findings of pulmonary cryptococcosis in King Chulalongkorn Memorial Hospital. The radiographs and computed tomography of the chests of the patients who had a diagnosis of pulmonary cryptococcosis, between 1998-2001, were retrospectively reviewed. Seven patients were included. Five were HIV infected, three had diffuse reticulonodular opacities, two (of the three) patients also had accompanying cavities; two had solely pleural effusion. Two patients were immunocompetent; one had a pulmonary nodule and another one had an endobronchial lesion and multiple pulmonary masses in the collapsed lung seen on CT scan, which were consistent with cryptococcoma. None had adenopathy. There was a difference in the radiologic manifestations between immunocompromised and immunocompetent hosts. Knowledge in radiographic features in pulmonary cryptococcosis should help radiologists to early recognize the disease and may improve the treatment outcome. PMID- 16471119 TI - A two-phase study model for the standardization of HER2 immunohistochemical assay on invasive ductal carcinoma of the breast. AB - OBJECTIVES: To develop and verify a standardized protocol for HER2 immunohistochemical assays on invasive ductal carcinoma of the breast in Thailand. MATERIAL AND METHOD: A two-phase study approach was employed. In the Phase One, after verifying the proposed protocol that adopted the HercepTest procedure using readily available primary antibodies, CB11 and A0485, Lab 1 performed the HER2 immunohistochemical staining for 137 cases of invasive ductal carcinoma twice with two types of the antibody. Nine pathologists from 8 centers independently examined and scored all the 2 x 137 stained slides that were blinded for antibody type. Interobserver reliability was calculated using pair wise kappa. Following discussion of the results, the Phase Two study was planned. Lab 2 and Lab 3 independently performed the HER2 staining according to the protocol for 60 invasive breast carcinoma cases. The same group of pathologists scored 2 x 60 stained slides that were masked for laboratories. Interobserver reliability and interlaboratory agreement from each pathologist were calculated using kappa statistics. Three interpreted categories--namely negative, equivocal and positive tests were used in the analyses. RESULTS: Phase One study showed interobserver agreement between pairs varied from kappa 0.75 (95%CI, 0.68-0.82) to 0.06 (95%CI, 0-0.14) while Phase Two study obtained pair-wise kappa scores ranged from 0.84 (95%CI, 0. 80-0.89) to 0. 65 (95%CI, 0.59-0.71). Interlaboratory kappa for each pathologist was 0.67 (95%CI, 0.61-0.73). CONCLUSION: The standardization of HER2 immunohistochemical assay was achieved through this two phase study model. It had added benefits of improving pathologists' expertise and verifying the HER2 testing protocol to be used in Thailand. PMID- 16471121 TI - Currarino syndrome: report of two cases and review of the literature. AB - In the present paper the authors examine two cases of Currarino syndrome and review the existing literature on the disease. Both cases presented with chronic constipation. The first concerns a two year old male born with anorectal stenosis and diagnosed with Currarino syndrome when scimitar sacrum and anterior meningocele were finally detected. The second concerns a 25 year old female who suffered from chronic constipation but was not diagnosed until thorough examination revealed ectopic anus with Hirschsprung disease, scimitar sacrum and anterior meningocele. Because these patients were not diagnosed with Currarino syndrome when first seen, the authors reviewed its prevalence, embryogenesis, clinical manifestations, diagnosis and treatment. The authors' research supports the significance of prompt diagnosis in effective treatment and reduction of morbidity. PMID- 16471120 TI - A predictive model for distinguishing ischemic from non-ischemic cardiomyopathy. AB - OBJECTIVES: To develop a predictive model to distinguish ischemic from non ischemic cardiomyopathy MATERIAL AND METHOD: The authors randomly assigned 137 patients with LV systolic dysfunction into two subsets--one to derive a predictive model and the other to validate it. Clinical, electrocardiographic and echocardiographic data were interpreted by blinded investigators to the subsequent coronary angiogram results. Ischemic cardiomyopathy was diagnosed by the presence of significant coronary artery disease from the coronary angiogram. The final model had been derived from the clinical data and was validated using the validating set. The receiver-operating characteristics (ROC) curves and the diagnostic performances of the model were estimated. RESULTS: The authors developed the following model: Predictive score = (3 x presence of diabetes mellitus) + number of ECG leads with abnormal Q waves--(5 x presence of echocardiographic characteristic of nonischemic cardiomyopathy). The model was well discriminated (area under ROC curve = 0.94). Performance in the validating sample was equally good (area under ROC curve = 0.89). When a cut-off point > or = 0 was used to predict the presence of significant coronary artery disease, the model had a sensitivity, specificity and positive and negative predictive values of 100%, 57%, 74% and 100%, respectively. CONCLUSION: With the high negative value of this model, it would be useful for use as a screening tool to exclude non-ischemic cardiomyopathy in heart failure patients and may avoid unnecessary coronary angiograms. PMID- 16471122 TI - Pseudoaneurysm of profunda femoris artery following internal fixation of intertrochanteric fracture: two cases report. AB - The authors report two cases of an uncommon but preventable complication after fixation of intertrochanteric hip fracture, the pseudoaneurysm of the profunda femoris artery. Both cases in the present study presented as proximal thigh mass, and soft tissue sarcoma was suspicious in one case. The diagnosis was confirmed by angiographic study and the feeding vessels were successfully occluded during angiography. Proper placement of the retractors, using a shorter drill or drill guard, accurate screw length and a shorter side-plate DHS were recommended to prevent this problem. PMID- 16471123 TI - Meningioma of the internal auditory canal: a case report. AB - Meningioma of the internal auditory canal is very rare. There are only 15 previous reports of intracanalicular meningioma. The authors add a case report of a patient with meningioma of the internal auditory canal. A 31-year-old woman presented with a one-year history of headache, dizziness, hearing loss and left facial paralysis. An MRI of the temporal bone demonstrated a tiny isointense intracanalicular tumor with inhomogeneous enhancement. In the operative field carried out by translabyrinthine approach, the tumor was found in the IAC without intracranial involvement. Pathology revealed a meningioma. The patient was followed up for 2 years without recurrence. PMID- 16471124 TI - Management on tsunami causing posttraumatic stress disorder: a case report. AB - On December 26, 2004, tsunamis hit Southeast Asia and caused serious damage and loss of lives. In Thailand, six provinces (Ranong, Phang-Nga, Phuket, Krabi, Trang, and Satun) were impacted. The present study reports the psychiatric assessments such as Thai GHQ-60 and IES. It also reports management techniques of both cognitive behavior therapy and medication. Those were provided to a Thai female patient who was 54 years old. The patient responded to treatment quickly because of early management. The tsunami victim with Posttraumatic Stress Disorder (PTSD) is not an individual. A mass of people who faced or witnessed the tsunami are vulnerable to get PTSD any time during 6 months after trauma. These early management techniques are useful and practical for a mass of victims and survivors. PMID- 16471125 TI - Auditory symptoms: a critical clue for diagnosis of MELAS. AB - Mitochondrial encephalopathy, lactic acidosis with stroke-like episodes (MELAS) is a rare mitochondrial disorder that affects adults. MELAS syndrome can mimic cerebrovascular disease, encephalitis or toxic-metabolic encephalopathy. The authors reported two patients who presented with auditory symptoms before the onset of encephalopathy and stroke-like episodes. The first patient was a 28 year old man, who presented with acute sensorineural hearing loss (SNHL) followed by headache, left hemiparesis and generalized tonic-clonic seizure. CT scan of the brain showed hypodensity lesion at the tip of right temporooccipital region. Audiogram and brainstem auditory evoked potential (BAEP) showed abnormal conduction of left brainstem auditory pathway. MRI of the brain showed a lesion involving gray and white matters of the right occipital, parietal and temporal lobes. The distribution of the lesions was not compatible with distribution of arterial supply. MRA was normal. The second patient was a 56 year-old woman with a one-year history of hearing loss. The audiogram revealed bilateral SNHL. A few days before admission, her hearing was acutely deteriorated She could not understand a conversation while she could communicate by writing. CT scan of the brain showed hypodensity in both temporal lobes and MRI revealed lesions in the same area. Pure tone audiogram showed moderate SNHL but BAEP was normal. One week later, she developed global dysphasia and generalized tonic-clonic seizure. Both patients had elevated cerebrospinal fluid and serum lactate: pyruvate ratio. Polymerase chain reaction-restriction fragment length polymorphism disclosed A3243G mtDNA mutation in the blood in the first patient and in muscle biopsy in the second patient. Ubiquinone supplement was prescribed The auditory symptoms in combination with stroke-like episode in supratentorium are important clues to diagnose MELAS syndrome. PMID- 16471126 TI - Acardiac twin. AB - Acardiac twin is a rare complication of monozygotic twin pregnancies occurring in 1% of monochorionic twin gestations with an incidence of 1 in 35,000 births. Without treatment, the perinatal mortality of the pump twin is about 50-70%. The optimal management of acardiac twin pregnancies is controversial. Management options include elective termination, observation with close antepartum surveillance (serial cardiotocography, ultrasonography and echocardiography) and surgical interventions PMID- 16471127 TI - Molecular mechanism of herbs in human lung cancer cells. AB - Herbs have been considered natural and valuable sources for anticancer drug discovery. Herbal medicine has been prescribed in many countries over centuries for treating various diseases including infectious and malignant diseases. Nowadays, many of the drugs that have been used for treatment of malignant diseases are derived from natural products such as Taxol, a natural product isolated initially from Pacific Yew (Taxus brevifolia). This review article describes research on molecular mechanisms, especially cytotoxic effect of natural products from plant sources, primarily preclinical studies, involving human lung cancer cells in vitro for providing more knowledge and issues for potential drug development from medicinal herbs in the future. PMID- 16471128 TI - GIS and epidemiology. AB - Understanding the spatial patterns of infectious diseases can provide insight as to their causes and controls. Geographic information systems (GIS) and related technologies like remote sensing are increasingly used to analyze geographical distribution of diseases as well as relationships between pathogenic factors (causative agents, patients, vectors and hosts) and their geographic environments. Basic and analytical applications of GIS in epidemiology can help in visualizing and analyzing geographic distribution of diseases through time, thus revealing spatio-temporal trends, patterns, and relationships that would be more difficult or obscure to discover in tabular or other formats. GIS can provide a means to meet the demands of outbreak investigation and response, where understanding the spatial spread and dynamics of an outbreak is central to the design of prevention and control strategies. PMID- 16471129 TI - The new medical training program in UK: modernising medical careers. PMID- 16471130 TI - [Mammography for breast cancer screening]. PMID- 16471131 TI - [European Union fight against smoking related activitiy]. AB - This study is aimed at providing information concerning the provisions adopted by the European Union on both a compulsory and non-compulsory basis for its member States related in one way or another to the fight against smoking. To this end, a review is made of all of the provisions published in the Official Journal of the European Union as of the first published in 1986 up to March 2005, commenting upon the aspects of each provision having to do with the subject stated above. PMID- 16471132 TI - [Differences in the factors related to the use of mammograms among Spanish women in 1994 and 2000]. AB - BACKGROUND: The screening mammography is the most highly effective procedure for early breast cancer detection. This study is aimed at ascertaining the differences in the factors related to Spanish women 40-70 years of age having had mammograms in 1994 and 2000 and among those having had at least one mammogram within the two years prior. METHOD: Comparison of the results of two surveys conducted in 1994 and 2000, each on samples of women representative of Spanish women 40-70 years of age (3,218 in 1994 and 2,409 in 2000). An analysis was made of sociodemographic, sociosanitary and the women's attitude and awareness-related variables. RESULTS: Statistically significant differences were found between 1994 and 2000, the intention of having a mammogram in the future (96% in 2000 vs. 79% in 1994) (p<0.0001). A total of 59% of the 51-65 age group had a mammogram in 2000, as compared to the 50% who had one in 1994. CONCLUSIONS: The profile of Spanish women has undergone a change from 1994 to 2000, although those women who had had a mammogram were of similar sociodemographic and sociosanitary characteristics. However, the screening is being done to a greater extent on the age 51-65 age group in 2000, and the women are showing more positive attitudes toward having a mammogram. PMID- 16471133 TI - [Impact on hospital days of care due to unnecessary emergency admissions]. AB - BACKGROUND: Unnecessary admissions of acute cases have major impacts on hospital efficiency and organization. This study is aimed to identify percentage of unnecessary admissions from a hospital emergency department and the reasons why, as well as to quantify the unnecessary hospital days of care generated by these admissions. METHODS: It has been analyzed the appropriateness of 622 admissions made in 2002, selected at random, all of the hospital stays generated by the inappropriate admissions and a representative sample of the appropriate admissions of a second-level hospital in Asturias. The review tool was the Appropriateness Evaluation Protocol. A descriptive analysis, a bivariate analysis and a multivariate logic regression analysis were made. RESULTS: A total of 63 admissions (10.1%) were judged inappropriate. The main cause of inappropriateness were admissions for performing diagnostic tests and/or treatments, which could be carried out on an outpatient basis. These unnecessary admissions generated 78.2% of unnecessary stays, and the appropriate admissions generated 24.8% of unnecessary stays. Referrals to hospital emergency rooms from physicians outside of the hospital proper heightened the risk of unnecessary admissions (OR:4.50, 95% CI: 1.59-12.76), daytime admissions (OR: 13.97, 95% CI: 1.86-104.76) or evening admissions (OR: 7.70, 95% CI: 1.01-58.72), admissions to cardiology wards (OR: 3.93, 95% CI: 1.22-12.70) and neurology wards (OR: 5.86, 95% CI: 1.88 18.30), the experience of prior admissions having lowered the risk of unnecessary admission (OR: 0.34, 95% CI: 0.18-0.65). CONCLUSIONS: Unnecessary admissions generate three times more inappropriate stays than the necessary admissions. Hospital organization-related problems are the main cause of inappropriate admissions. PMID- 16471134 TI - [Variability in primary care drug prescription in 2003 in Castile-la Mancha, Spain]. AB - BACKGROUND: Detecting variability in clinical practice is important, given that it may entail the underuse or overuse of resources. This study was aimed at studying the variability of drug prescription in Primary Care in Castile-La Mancha in terms of certain sociosanitary characteristics of each health district. METHODS: The prescribing of various therapeutic groups in 2003 was reviewed in 180 health districts, the following variables having been gathered: number of packages having been prescribed per 100 inhabitants, percentage of inhabitants over age 65 in the district, visit rate (visits/1,000 inhabitants), physicians/1,000 inhabitants and distance to the reference hospital. RESULTS: The greatest degree of drug-prescribing variability was found regarding fibrates (42.95% variance factor) and H2-blockers (38.61%). The group showing the closest correlation between its prescription rate and the variables analyzed was that of the angiotensin-converting enzyme inhibitors (mean Spearman p: 0.719); antidepressants however showing a weak correlation (0.324). The percentage of inhabitants over age 65 and the visit rate are the variables arising more significantly in the multiple regression models constructed for each treatment group. The highest determination coefficient was found in the angiotensin enzyme converting inhibitors (R2=0.761). The mean determination coefficient for all of the groups, weighted on the basis of the prescribing volume, was 0.492. CONCLUSIONS: A high degree of drug-prescribing variability was found to exist according to the different districts. Generally speaking, the variables analyzed explain part of this variability. PMID- 16471135 TI - [Prevalence of physical activity and its relationship to sociodemographic variables and lifestyles in the age 18-65 population of Pamplona, Spain]. AB - BACKGROUND: Current scientific knowledge upholds the possibility of considering a cause-effect relationship to exist between the degree of physical activity and the onset of different diseases. Des pite this, physical activity is not a very well-rooted practice among the population, which is progressively becoming more sedentary. This study is aimed at describing the prevalence of sedentarism among the population of Pamplona, as well as ascertaining what factors have the greatest bearing thereon. METHODS: A cross-sectional study was conducted by means of a postal survey on a significant sample of the age 18-65 population of Pamplona. The survey used included an evaluation of the degree of physical activity based on the Paffenbarger physical activity questionnaire. A bivariate and multivariate analysis was made using logic regression. RESULTS: A total of 76.6% of the females have a sedentary lifestyle, whilst this percentage drops to 56.7% among males. Sedenta rism increases with age, up to 80.3% of males and 86.3% of females being respectively sedentary in the oldest age group. CONCLUSIONS: The sociodemographic factors such as sex, age, formal education, occupation and marital status seem to be determi ning factors of the sedentary lifestyle, while smoking and being overweight or obese are not. Among the young, the females not having a college education and married men who are smokers seem to be target populations for promoting physical activity. PMID- 16471136 TI - [Characteristics of the poisoning mortality in the 1986-2001 period in the Autonomous Basque Community, Spain]. AB - BACKGROUND: Acute drug-related deaths are frequent, giving rise to high Potential Years of Life Lost figures. The objectives of this research are to ascertain the trend in the acute drug-related death rate in the Autonomous Basque Community and the variance thereof in terms of different variables. METHOD: The drug-related death rate data from the Autonomous Basque Community's Death Register for the 1986-2001 period was analyzed overall in terms of different demographic variables and by cause of death (ICD-9 and ICD-10). The analysis includes gross death rates and standardized rates by European population. The potential years of life lost are calculated. RESULTS: Acute drug-related deaths numbering 1,207 in all were recorded during the period under study, totaling 045% of all deaths, for a gross rate of 3.58 deaths/100,000 inhabitants. The mean Potential Years of Life Lost were 2,226.33/year, for a rate of 1.12/1000. A total of 75.97% of these deaths were males, the average age at time of death having been 40.29 years (36.09 for males and 52.64 for females). The etiology most often involved was: accidental (82.19%), followed by intentional (12.43%) and undetermined (5.38%)-. Accidental poisoning by other drugs (E-850 in ICD-9, X44 in ICD-10) was the leading cause of death (42.30% and 34.75% respectively). CONCLUSIONS: There has been an increase in the number of acute drug-related deaths throughout the period studied, the accidental prescribed drug overdose being the main cause, entailing statistically significant differences by sex and age. PMID- 16471138 TI - [Psittacosis outbreak in Granada, Spain]. AB - BACKGROUND: Psittacosis can be transmitted to humans from infected birds, being the cause of atypical pneumonia. This work is aimed at determining the source of a psittacosis outbreak, the characteristics of the subjects exposed and describing the measures to control the outbreak. METHODS: Descriptive study. Notification to the Epidemiological Monitoring System and Autonomic Agriculture authorities, taking the pertinent measures (inspection, sampling, disinfecting and precautionary quarantine of birds), closing the establishment. Case definition and epidemiological survey. Frequency analysis. RESULTS: Seventeen (17) individuals (one of whom died) came into contact with suspicious animals (53% males/47% females), nine of whom (mean age: 30) showed symptoms (100% fever, flu-like symptoms 44.4%). A serology was performed for C. psitacci on 11 of the 17 subjects exposed (68.75%), two of whom (18.18%) tested positive. The presence of C. psittaci DNA was detected in the pulmonary tissue study conducted on the deceased individual. A total of 33% (3/9) of those showing symptoms were confirmed by diagnosis. The Autonomic Agriculture authorities placed 22 animal compounds under quarantine. A total of seven (10%) of the 70 samples taken (60: animal cloacas and 10 birds) revealed a positive antigen, the PCR nucleic acid test being positive in one there-of. CONCLUSIONS: The exposure to diseased animals at the establishments involved caused the outbreak, the common source of contagion being of an occupational nature. The appropriate coordination/collaboration between the local Health and Agriculture authorities made it possible to prevent any further cases from arising. PMID- 16471137 TI - [Educational intervention for preventing unwanted pregnancies and sexually transmitted diseases among teenagers in the city of Toledo, Spain]. AB - BACKGROUND: No-one doubts the need of effectively providing teenagers with information about birth control and sexually-transmitted diseases. This study is aimed at evaluating the results of an educational intervention related to these matters. METHODS: Before-and-after study of an educational intervention (based on lectures and handing out documentation) without a control group. A questionnaire was passed out before and after the intervention to assess changes in knowledge and attitudes of the 4th-year Compulsory Secondary Education students at five schools in Toledo. RESULTS: The questionnaire was answered by 238 of the 268 students. The average age was 15.59. A total of 54.66% were females. In all, 24.03% had had some sexual relation. The birth control method used most often was the condom (98.24%). The girls more refuse more unprotected relations (76.5% vs. 48.6%; p<0.001) and share the same classroom with a student having AIDS (80.47% vs. 60.38%; p<0.001). Six months following the start of the intervention, a total of 197 students answered the second questionnaire. Proper condom use rose from 62.13% to 73.46%. CONCLUSIONS: Following the intervention, an improvement has been noted in the degree of knowledge related to birth control methods and AIDS transmission and a more positive attitude regarding HIV. PMID- 16471139 TI - [Incidence of pneumococcal pneumonia: the bias of information source]. PMID- 16471140 TI - Late fertility in Europe: some comparative and historical data. AB - THE ISSUE: Given the substantial increase in late fertility (at ages 35 and older) in the last few years in Western countries, the paper raises the question of what factors have caused this new trend, and how likely it is to continue. METHODS: Late fertility is measured using the sum of age-specific fertility rates at 35 or more; its contribution to the total fertility rate (in % of the total fertility rate) and the proportion of first childbirths in this fertility is then computed. Late fertility among men is assessed using the same method, where the necessary data are available. The data analysed sometimes date back to the 18th century for women (Sweden), and the early 20th century for men (in France). RESULTS: Late fertility is now much lower than it used to be. Despite the current increase--which can be traced back to the early 1980s for most Western countries, and appears to be much more recent and limited in the former socialist countries- current levels are very low compared with those of the past, among men as well as women. Reduced family size was the main factor in that decline, which became more pronounced as a result of women having a first child at younger ages. The reversal of the trend is due only to postponed childbearing, which must be viewed as part of a general trend towards postponement of all demographic events. More and more of the children born in such circumstances are thus first children. Very late fertility (at 40 or more) has increased, though only slightly, for men as well as women, and age 45 seems to be a limit beyond which few are prepared to have children. Social norms concerning the age limit for a person to have a child could account for the differences from one country to another in that respect. CONCLUSION: Late fertility is likely to continue increasing as long as fertility continues to decrease among young people. However, the increase will continue to be due mostly to the "younger" age groups (under 40 for women, under 45 for men); both biological constraints and social norms probably play a part in the stagnation of very late fertility levels. PMID- 16471141 TI - Who are the late mothers? AB - BACKGROUND: The frequency of late births (to mothers aged 35 or above) has increased since the 1980s, because the number of women over this age has increased (baby-boomer generation) and above all because their fertility has increased. The characteristics of late mothers have changed in the last 20 years: more primiparous women, more women in second unions, more women with a high educational level etc. Some of these trends are related to a general shift in the characteristics of the population, whereas others are the consequence of a change in women's fertility after 35 for some specific categories of women. METHODS: We compared the characteristics of "late births" with those of births to mothers aged below 35 and with those of all women aged over 35. Fertility among several groups of women over 35 and the proportion of "late births" among different groups of births offer different insights into trends according to birth order, mother's marital history, level of education and occupation. RESULTS: Late births are more often of order 1 because the fertility of childless women over 35 has increased substantially. Conversely, more births are to women living in a second union, mostly because second unions are becoming more common. Late mothers are more educated than before. On the one hand, the overall level of education has increased. On the other hand, highly educated women are more likely to be childless and without a partner at age 35, and fertility after 35 is now highest for childless women living in a couple. Late mothers are increasingly likely to have middle-level or higher-level occupations, because they rarely have children before reaching such positions. CONCLUSION: Late mothers are now more numerous than 40 years ago and their mean social status has improved. This may change the way in which age is considered to be a risk factor for pregnancies: late mothers are more often primiparous, but their pregnancies are now more planned than before. PMID- 16471142 TI - Age and male fertility: biological factors. AB - Although the effect of maternal age on fertility is well known, whether paternal age also affects fertility remains unclear. If there is an effect, the biological causes and consequences, especially for the offspring, are unclear. We present a short review of the data and various studies on this subject. Data on young men or fathers are not considered here. Advanced male age has been associated with: Increased time-to-pregnancy (TTP) and subfecundity; low semen volume, total sperm count, sperm motility and normal sperm morphology; gradual modification of testicular vascularisation and regular decrease in the number of Sertoli and Leydig cells, with an associated decrease in serum testosterone. There are wide interindividual variations of these modifications, with some elderly men having a reproductive function similar to young men. Whether there is an increase in levels of qualitative and or quantitative alterations of the sperm chromosomes with advancing age remains controversial. It can be suggested that there is an increased risk of developmental defects in the offspring of older men, as the spontaneous mutation rate is closely related to the age of the man. An increasing rate of miscarriages has been related to older fathers and several studies suggest older fathers are at the origin of several diseases in the new-born. In conclusion, further studies are needed due to the increasing number of older men wishing to become a father. PMID- 16471143 TI - The biological aspects of risks of infertility due to age: the female side. AB - When women reach their forties, and despite strong inter-individual variations, their fecundity, either spontaneous or assisted, is impaired then falls down from the age of 45 onwards. In humans the size of the ovarian reserve is definitely fixed at birth and decreases drastically with age. Follicles leave the stock of resting follicles in a continuous stream, either disappearing by apoptosis or entering the growth phase. From the age of about 38 years onwards, the rate of follicular depletion accelerates leading to a decreasing number of selectable follicles. As these follicles produce inhibin B, its circulating levels drop, resulting in an increase in circulating FSH concentration. The higher rate of growth of large follicles in women over the age of 40 than in younger women, likely due to high FSH levels, and/or an early selection of the preovulatory follicle seem responsible for the shortening of the follicular phase in women in their late thirties and early forties. An impaired endometrial receptivity and/or a decrease in oocyte quality, or both may be responsible for the increase of the frequency of spontaneous abortion when women reach their forties. In conclusion, it can be predicted that, although apparently inescapable, the fertility impairment due to ageing could be, at least partly, counteracted by the use of molecules preventing attrition of the ovarian reserve. PMID- 16471144 TI - Paternal age: are the risks of infecundity and miscarriage higher when the man is aged 40 years or over? AB - BACKGROUND: Maternal age of 35 years or over is a well-known risk factor for human reproduction that has been extensively investigated by demographers and epidemiologists. However, the possibility of a paternal age effect has rarely been considered. We carried out review of the literature to investigate the effect of paternal age on the risks of infecundity and miscarriage. METHODS: We carried out a MEDLINE search and checked the exhaustiveness of our reference list. RESULTS: We identified 19 articles analysing the effect of paternal age. Epidemiological studies provided evidence that paternal age older than 35-40 years affects infecundity. However, the few studies based on data from assisted reproductive techniques (especially IVF with ovum donation) do not confirm this finding. All studies analysing the effect of paternal age on the risk of miscarriage showed an increased risk in men aged 35-40 years or over. Other studies have shown some evidence for a paternal age effect on late foetal deaths. CONCLUSION: The risks of infecundity and miscarriage increase with paternal age. Two main hypotheses can be considered. First, these risks increase after the age of 35-40 years. However, a later paternal age effect (after 45-50 years) cannot be excluded. Second, due to the interaction of the ages of the two partners, the risks of infecundity and miscarriage may be higher when both partners are older (woman aged 35 years or over and man aged 40 years or over). PMID- 16471145 TI - Age effect: results from a detailed prospective study on daily fecundability. AB - BACKGROUND: Demographers, epidemiologists and clinicians have long been interested in the estimation of age-specific fecundability. With the progressive postponement of age at family formation in Western countries, this topic has recently become the focus of renewed attention. METHODS: In order to correctly estimate the effect of biological ageing on fecundability it is crucial to collect detailed information through a rigorous study design and to apply suitable models for analysing time to pregnancy data. In this article we discuss some methodological problems concerning the study of fecundability and its dependence on biological ageing. We also present the Multinational Study in Daily Fecundability which has produced a very rich database with detailed behavioural and physiological information prospectively collected on every cycle. Finally, we review some results on age effects obtained from these data. RESULTS: Our findings show that the decline in fecundability from age 28 to age 33 is not statistically significant and very modest. CONCLUSION: The results presented here do not appear to be particularly alarming with regard to the postponement of conception of the first child from age 28 to 33. This postponement could however lead to attempts to conceive a second child after age 35, when fecundability starts to decrease rapidly and when the risk of genetic disorders increases. PMID- 16471146 TI - Controlling for continuous confounding factors: non- and semiparametric approaches. AB - Confounding is one of the major types of bias encountered in observational epidemiologic surveys designed to study the relation between an exposure factor and a health event. A common way to remove confounding bias during the statistical analysis phase is to adjust for the confounders in a regression model. If a confounding factor is assessed as a continuous variable, it is necessary to define how the variable is entered into the regression model. In the case of logistic regression, we illustrate through simulation that coding by a binary variable or a categorical variable with broad categories may lead to substantial residual confounding. Specific approaches can be used to define a coding method that limits residual confounding. Among these, we briefly present nonparametric approaches and describe in detail several semiparametric approaches (generalised partial linear models, spline regression and fractional polynomials). These can be used to estimate the relation between a continuous factor and the health event of interest by a smooth non pre-specified function. In semiparametric models, the effect of certain covariates is coded by a parametric function, whereas the coding of one or two continuous variables is represented by a nonparametric function. These models can be used in exploratory analyses to describe dose-effect relations between the confounder and the health event, and thus help to define a relevant coding for the confounder. PMID- 16471147 TI - Foetal mortality, infant mortality, and age of parents. An overview. AB - This review article examines the relationship between late foetal and infant mortality, and age of parents. The highest risks are observed at older maternal ages for foetal mortality and at both extremes of reproductive ages for infant mortality. For infant morbidity, the role of intermediate variables is discussed. Increasing paternal age seems to be related to higher foetal and neonatal mortality. PMID- 16471149 TI - Late childbearing and its impact on adverse pregnancy outcome: stillbirth, preterm delivery and low birth weight. AB - BACKGROUND: The role of parental ageing on the incidence of adverse pregnancy outcome is based on increased morbidity and obstetric problems during pregnancy and delivery in old mothers, and on the accumulation of spontaneous harmful mutations for continuous cell divisions during spermatogenesis in old fathers. The aim of this study is to estimate the impact of paternal and maternal ageing on the risk of adverse pregnancy outcome. DATA AND METHODS: From the group of 3,616,622 Italian singletons born in 1990-1996 we estimated the risks of stillbirth, preterm birth (<37 weeks of gestation) in live births, and low birth weight (< 2.3 Kg) in live full-term births. The risks were estimated as a function of maternal and paternal ageing through logistic regression models, which included, as covariates, parity (1st, 2nd, > or =3rd) and family education (low, < or =8 years of schooling for both parents; high, >8 years for at least one parent). Parental ages were examined as quantitative (in one year classes) or categorical factors (in three classes: fathers 20-29, 30-39, > or =40; mothers 20 29, 30-34, > or =35). RESULTS AND CONCLUSIONS: We found that, compared with 20-29 year old parents, mothers > or =30 years and fathers > or =40 years are at risk of adverse pregnancy outcome. The effects are more relevant for preterm births and greater in the least than in the most favourable birth conditions, i.e., in first-born children of less educated families than in second-born children of highly educated families. For the risk of a preterm delivery, the odds ratio is OR = 1.32 [1.28-1.36] in mothers aged 30-34 years, and OR=1.97 [1.88-2.07] in mothers 235 years in the least favourable conditions, and OR = 1.14 [1.10-1.18] and OR = 1.56 [1.22-1.27] respectively, in the most favourable conditions. The impact of paternal ageing is smaller but significant in fathers > or =40 years: for the risk of a preterm birth, the odds ratio is OR = 1.40 [1.33-1.47] in the least favourable conditions, and OR = 1.14 [1.08-1.21] in the most favourable conditions. This last, baseline risk might be indicative of a paternal genetic component associated with childbearing in advanced age. PMID- 16471148 TI - Trends and geographic inequalities in the prevalence of Down syndrome in Europe, 1980-1999. AB - BACKGROUND: EUROCAT is a network of population-based registries for the epidemiologic surveillance of congenital anomalies covering approximately one quarter of births in the European Union. Down syndrome constitutes approximately 8% of cases of registered congenital anomaly in Europe, with over 7000 affected pregnancies in the 15 current member states of the European Union each year. In this paper, we aim to examine trends in the live birth prevalence of Down syndrome in Europe in the light of trends in maternal age and in prenatal diagnosis. METHODS: Descriptive analysis of data from 24 EUROCAT registries, covering 8.3 million births 1980-99. Cases include live births, stillbirths and terminations of pregnancy following prenatal diagnosis. RESULTS: Since 1980, the proportion of births to mothers of 35 years of age and over has risen quite dramatically from 8 to 14% for the European Union as a whole, with steeper rises in some regions. By 1995-1999, the proportion of "older" mothers varied between regions from 10% to 25%, and the total prevalence (including terminations of pregnancy) of Down syndrome varied from 1 to 3 per 1000 births. Some European regions have shown a more than twofold increase in total prevalence of Down syndrome since 1980. The proportion of cases of Down syndrome which were prenatally diagnosed followed by termination of pregnancy in 1995-1999 varied from 0% in the three regions of Ireland and Malta where termination of pregnancy is illegal, to less than 50% in 14 further regions, to 77% in Paris. The extent to which terminations of pregnancy were concen trated among older mothers varied between regions. The live birth prevalence has since 1980 increasingly diverged from the rising total prevalence, in some areas remaining approximately stable, in others decreasing over time. CONCLUSION: The rise in average maternal age in Europe has brought with it an increase in the number of pregnancies affected by Down syndrome. The widespread practice of prenatal screening and termination of pregnancy has in most of the regions covered by EUROCAT counteracted the effect of maternal age in its effect on live birth prevalence. Under the joint influences of maternal age and prenatal screening the pattern of geographic inequalities in Down syndrome live birth prevalence in Europe has also been changed. PMID- 16471150 TI - Heterogeneity, the masked part of reproductive technology success rates. AB - BACKGROUND: Heterogeneity in women and men's fecundity is a well-established fact. The selection process of men and women treated for infertility might bias the success rates of reproductive technology. Bias may also arise from frequency and timing of intercourse with respect to the day of ovulation. METHODS: Several datasets were collected and analysed. They concern normally fertile couples who used natural family planning methods and infertile couples treated by artificial insemination with husband or donor's spermatozoa. The effects of heterogeneity on the success rates of treatment cycles are described and solutions are proposed as to data collection and statistical analysis in this specific field. RESULTS: The decrease in the success rates along successive cycles of assisted reproduction is a consequence of heterogeneity. The probability of conception varies among women and among men. After a first success, the probability of another success is higher. After a failure, the probability of success is lower. The most specialised centres treat the less fecund couples. There is a negative correlation between men and women's fertility in case of oligo/azoospermia. The fecund window cannot be correctly located by calendar calculations, but more appropriately by assessment of cervical mucus at the vulva. The variability of this location is wide. The decrease in the success rate with men and women's age results from a complex mixture of an increase in the proportion of sterile patients and a decrease in fecund patients' fecundity. CONCLUSION: Care should be taken to limit the bias due to patient selection and specific statistical methods should be used to allow for the progressive selection of patients during fertility studies and for the variability of the frequency and the timing of intercourse or insemination relative to ovulation. PMID- 16471151 TI - How effective is assisted reproduction technology? A model assessment. AB - BACKGROUND: All conceptions occurring after a treatment for infertility are usually attributed to the treatment. This is only true when only totally sterile couples are treated: subfecund couples may still conceive without treatment. METHODS: We used a computer simulation (Monte Carlo) model of reproduction, combining the monthly probability of conceiving, the risk of miscarriage and the probability of age dependent permanent sterility. After a delay (D) without conceiving, the couples are assumed to turn to assisted reproduction technology. The treatment increases the fecundability (monthly rate of conception) of subfecund couples and restores the fertility of some totally sterile couples. The overall effectiveness of the therapy is based on data (by age of women) for two attempts of IVF. The number of conceptions ending in a live birth with ART is compared to the number of conceptions after one or two additional years without ART. RESULTS: At 30 years of age, the crude effectiveness of ART, based on all conceptions after treatment, is double that observed without treatment (30% vs. 15%), and beyond 41 years of age, the number of conceptions with treatment is no higher than without treatment over two years of exposure to conception (15% for both at 41 years of age, 7% at 45 years of age). Shortening the delay (D) does not improve the total number of women conceiving and having a live birth unless we assume the couple try again to spontaneously conceive. CONCLUSIONS: The chances of conceiving spontaneously must not be ignored when defining an infertility treatment, especially in cases of unexplained infertility. Shortening the delay before using ART is not in itself a more efficient strategy. PMID- 16471152 TI - Purpose, development and use of injury indicators. AB - Injury indicators can be used to give policy makers an estimate of the scale of injuries and their long-term effects. They can help compare injury levels in different areas and countries and can be used to help measure the effectiveness of interventions. Work on severity related indicators is promising. However there are no perfect indicators to date as many are hampered with difficulties in case definition and under reporting. For example, mortality rates are affected by improvements in care even if the incidence of an injury remains the same, the abbreviated injury scale (AIS) takes 10-20 minutes to code and so is not used in health service databases, surveys have problems with recall bias, definition of injury and response rates. If we accept that we need to make the best out of imperfect indicators and imperfect data then we should use multiple sources of data and accept that no one indicator can be used universally but needs to be selected for the purpose. For example, one possible new indicator of the incidence of non-fatal injury might be fracture data in the emergency department. Fractures are painful and so nearly always end up with a hospital attendance. This might give a means to compare incidence of non-fatal injury in different areas and countries. In conclusion, we need injury indicators to progress in injury prevention. Imperfect indicators can be used for targeting and evaluating interventions as long as we know and adjust for their limitations. PMID- 16471153 TI - Indicators of injury burden: which types are the most important? AB - Injury indicators are used for monitoring the impact of injury prevention initiatives on the population burden of injury. The object of the present study was to identify the types of injury responsible for the major component of the population health burden of injury in a large cohort in Manitoba, Canada. Injury cases (ICD-9-CM 800-995) aged 18-64 years were identified from all Manitoba hospital data between 1988 and 1991. Morbidity data were obtained from hospital discharge abstracts 12 months prior to date of injury and for 12 months post injury. Outcomes for individuals were calculated as the difference pre- and post injury in hospital inpatient days. Death outcomes in the 12 months post-injury were obtained by linking the cohort with the population registry. Summed outcomes across the population were stratified into injury types based on the International Code of Diseases (ICD) code of the index injury. Outcomes were also stratified by injury severity score categories where the injury severity score was obtained using ICDMAP-90. When ranked by contribution to the cohort's cumulative hospital inpatient days in the 12 months post-injury, the six most common ICD subchapter groups accounted for 65% of the total inpatient days. These six injury types also accounted for 62% of the total number of deaths in this cohort in 12 months after injury. The suggested injury types to use as indicators of burden include fracture of the lower limb, fracture of the head and neck, poisonings, intracranial injury, fracture of the upper limb, and fracture of skull. PMID- 16471154 TI - Injury outcome indicators--validation matters. AB - INTRODUCTION: There is concern that many national non-fatal injury indicators currently in use are misleading. OBJECTIVE: To make the case for the validation of existing unvalidated indicators, as well as the validation of new indicators before they are promulgated. METHOD: The International Collaborative Effort on Injury Statistics (ICE) Criteria were used for investigating the validity of indicators. Examples of indicators that have been found to be valid using these criteria are presented. In contrast, examples of national road safety indicators are also presented, whose validity is questionable. Trends in road safety indicators with and without threats to validity are contrasted. RESULTS: The New Zealand Injury Prevention Strategy (NZIPS) serious injury indicators are presented as indicators with no identifiable threats to validity. National road safety indicators from Canada, New Zealand and the United Kingdom, with identifiable threats to validity, are also presented. When trends for the valid NZIPS motor vehicle traffic crash indicators are compared with the New Zealand national road safety indicators, which have identifiable threats to validity, they show contrasting trends. This raises concerns that the current national indicators are potentially misleading. CONCLUSION: Validation does matter. For any indicator, it is important that it is clearly defined and specified. The specification should make it clear what parameter the indicator aims to reflect. Before use, the indicator should be validated against this target parameter. That parameter, and the indicators aimed to estimate it, should focus attention on important injuries, ie. injuries that are associated with significant mortality, threat-to-life, threat-of-disablement, loss of quality of life, or increased cost. PMID- 16471155 TI - An evaluation of the assessment of quality of life utility instrument as a measure of the impact of injury on health-related quality of life. AB - The Assessment of Quality of Life (AQoL) is a generic health-related quality of life (HRQL) measure. It is the only HRQL instrument, currently available, that incorporates health preference values derived from an Australian population and has been extensively trialled in over 40 studies. However, prior to this study, it had not been used to measure HRQL in injury patients. The aim of this study was to evaluate the AQoL, as a measure of the impact of injury on HRQL, by examining its correlation with other commonly used measures of health outcome and its ability to discriminate between groups with injuries of varying type and severity. A total of 221 admitted injury patients, aged 18-74 years, were recruited into the study from four major Victorian metropolitan hospitals and followed up over 12 months. The AQoL and the SF-36 were administered to obtain retrospective measures of pre-injury HRQL and health status with post-injury measurements obtained at five intervals post-injury (to 12 months). A preliminary analysis of data from this study showed the AQoL was positively related to other common outcome measures and, overall, showed a strong correlation with the SF-36 Physical Component Summary and a moderate correlation with the Mental Component Summary. It also demonstrated good discrimination between groups on the basis of type of injury, body region injured and severity of injury. While further testing of the AQoL, in this context, is still necessary, this study suggests that the AQoL may be a useful measure of the impact of injury on HRQL. PMID- 16471156 TI - How well do anatomical-based injury severity scores predict health service use in the 12 months after injury? AB - There is an acknowledged need for valid and reliable injury scores, suitable for use at the population level, which can accurately predict the long-term outcome of injury. The objective was to quantify the extent to which the abbreviated injury severity score (AIS) and the functional capacity index score (FCI) predict use of health services in the 12 months following an injury event. A cohort of injured people (ICD-9-CM 800-995) aged 18 - 64 years was identified from Manitoba hospital discharge abstracts from January 1988 to December 1991. For each member of the cohort whose injuries could be mapped to an abbreviated injury scale unique identifier, a maximum AIS (maxAIS) and a maximum FCI (maxFCI) were obtained. The cohort was linked with hospital discharge abstracts, physicians' claims and deaths from the population registry for the 12 months following injury. Negative binomial regression was used to model the relationships between the severity scores and the three outcome measures, while controlling for potential confounding variables. In total, 20 677 (97%) eligible cases were identified, of which 16 834 (81%) could be assigned a maxAIS and 15 823 (77%) a maxFCI. MaxAIS and maxFCI were significantly associated with total days in hospital following injury, but explained little of the variation in any of the health service use outcome variables (maxAIS, partial pseudo r2 ranging from < 0.001 to 0.041; and maxFCI, partial pseudo r2 ranging from < 0.001 to 0.018). It was concluded that anatomical damage is only partly responsible for long-term injury outcome. Additional variables would need to be included in predictive models of health outcomes of injury before these models could be reliable. PMID- 16471157 TI - Making evaluation an integral part of injury programme implementation. AB - Uncertainty about the nature of evaluation can lead to injury prevention programme evaluations being poorly resourced and poorly conducted. The aim of this paper is to demystify programme evaluation and, by stressing its function, offer a perspective on evaluation that may encourage its more widespread integration in the general activity of injury prevention and control. Programme evaluation is best understood simply as the process of getting answers to essential questions about a programme. Methodology used in evaluation needs to be based in empirical science but is otherwise unrestricted except by the chosen question and the practical circumstances relating to the programme and the community in which it is implemented. Discussion about which methodology is appropriate for evaluation research is (unwittingly) a debate about 'which questions should you be asking?'. If the right people ask the right (and properly formulated) questions and build the means of obtaining the information to answer these questions (using appropriate methodology) into the conduct of the programme then evaluation will no longer be a problem but an essential component of the overall effort to reduce the community burden of injury. PMID- 16471158 TI - Designing the European Road Safety Observatory. AB - The unavailability of consistent traffic accident data and road safety information limits the opportunities to provide target approaches to reduce road crashes. The European commission has decided to meet the demand for this data by establishing a new Road Safety Observatory. The structure and much of the initial content is being developed within the SafetyNet Integrated Project. This paper describes the structure of its key components. PMID- 16471159 TI - The Functional Capacity Index, second revision: morbidity in the first year post injury. AB - As the field of injury outcomes measurement has moved beyond counting deaths and injuries, the need for instruments to measure long-term outcomes has increased. Early validation efforts indicated that the Functional Capacity Index (pFCI12) was a promising tool for predicting functional loss 12 months post trauma, but that the predictive validity for lower extremity fractures was low. The pFCI12 was recently revised in conjunction with the 2005 revision of the Abbreviated Injury Scale (AIS). This preliminary study was undertaken to assess the performance of the revised pFCI12 in predicting outcomes for patients with lower extremity trauma. This case review study used the Crash Injury Research and Engineering Research (CIREN) database which includes AIS codes, extensive clinical data and 3- and 12-month outcomes. Generally predicted functional loss at 12 months was greater with the pFCI12 (AIS 2005) than with the original FCI based on AIS 1990. All patients whose pFCI12 scores predicted functional loss experienced some morbidity at 1 year. The pFCI12 appears to provide improved predictions of functional outcome 12-months post-trauma for persons experiencing lower extremity trauma. PMID- 16471160 TI - Scotland: meeting the challenges. PMID- 16471162 TI - Northern Ireland: 2006 means excitement ahead. PMID- 16471161 TI - Wales: making connections. PMID- 16471163 TI - England: progress made, more to be done. PMID- 16471164 TI - Beating abuse. PMID- 16471165 TI - Ask the experts: spirituality in care. PMID- 16471166 TI - From theory to practice: capturing the uniqueness of nursing. PMID- 16471167 TI - Evaluation of an integrated falls education group programme. PMID- 16471168 TI - Anxiety and depression. AB - This article explores anxiety states among older people and how they may overlap with depression. It discusses prevalence, symptoms and treatment, and aims to help nurses to develop appropriate intervention strategies and to know when they should refer patients to other health practitioners. PMID- 16471169 TI - Taking tympanic temperature. PMID- 16471170 TI - Empowering the people. PMID- 16471171 TI - Blood pressure-lowering effects of lifestyle modification: possible involvement of nitric oxide bioavailability. AB - Lifestyle modification is recommended as a non-pharmacological approach to treatment of hypertension. Many investigators have reported that exercise has antihypertensive effects, and various mechanisms have been proposed to explain this phenomenon. For example, nitric oxide (NO), which may be increased by exercise, has been reported to play a crucial role in preserving vessel homeostasis both by regulating vascular tone and by exerting anti-atherosclerotic effects. NO is known to be exquisitely sensitive to inactivation by superoxide radicals. However, the relationship between the blood pressure-lowering effect of lifestyle modification and NO bioavailability remains unknown. We investigated the effects of a 12-week lifestyle modification program consisting of mild exercise and diet on changes in blood pressure, plasma nitrate/nitrite (NOx), plasma nitrotyrosine, which is the footprint of NO interaction with reactive oxygen species, and plasma extracellular-superoxide dismutase (EC-SOD). The 12 week lifestyle modification program lowered blood pressure and increased plasma NOx. When the subjects were divided into two groups according to the change of plasma nitrotyrosine as an indicator of NO bioavailability, the subjects whose plasma nitrotyrosine decreased exhibited a significant relationship between the blood pressure-lowering effect of the lifestyle modification and the increase in EC-SOD, whereas those without a decrease in plasma nitrotyrosine exhibited a significant relationship between the blood pressure-lowering effect and the increase in maximum oxygen consumption. These results indicate that the level of NO bioavailability influences the mechanism of the blood pressure-lowering effect of aerobic exercise and diet. PMID- 16471173 TI - Association between carotid hemodynamics and asymptomatic white and gray matter lesions in patients with essential hypertension. AB - The aim of this study was to clarify the magnitude of common carotid artery (CCA) structural and hemodynamic parameters on brain white and gray matter lesions in patients with essential hypertension (EHT). The study subjects were 49 EHT patients without a history of previous myocardial infarction, atrial fibrillation, diabetes mellitus, impaired glucose tolerance, chronic renal failure, symptomatic cerebrovascular events, or asymptomatic carotid artery stenosis. All patients underwent brain MRI and ultrasound imaging of the CCA. MRI findings were evaluated by periventricular hyperintensity (PVH), deep and subcortical white matter hyperintensity (DSWMH), and etat crible according to the Japanese Brain dock Guidelines of 2003. Intima media thickness (IMT), and mean diastolic (Vd) and systolic (Vs) velocities were evaluated by carotid ultrasound. The Vd/Vs ratio was further calculated as a relative diastolic flow velocity. The mean IMT and max IMT were positively associated with PVH, DSWMH, and etat crible (mean IMT: rho=0.473, 0.465, 0.494, p=0.0007, 0.0014, 0.0008, respectively; max IMT: rho=0.558, 0.443, 0.514, p=0.0001, 0.0024, 0.0004, respectively). Vd/Vs was negatively associated with etat crible (rho=-0.418, p=0.0038). Carotid structure and hemodynamics are potentially related to asymptomatic lesions in the cerebrum, and might be predictors of future cerebral vascular events in patients with EHT. PMID- 16471172 TI - Impact of the angiotensin II receptor antagonist, losartan, on myocardial fibrosis in patients with end-stage renal disease: assessment by ultrasonic integrated backscatter and biochemical markers. AB - Myocardial fibrosis commonly occurs in patients with end-stage renal disease (ESRD) and has proven to be an important predictor for cardiovascular events. In experimental settings, angiotensin II type 1 receptor (AT1-R) antagonists have been shown to have anti-fibrotic effects on the myocardium independent of their antihypertensive effects. In this study, to investigate whether the AT1-R antagonist losartan would have such anti-fibrotic effects in patients, we administered losartan or, for purpose of comparison, the angiotensin-converting enzyme enalapril or Ca2+-antagonist amlodipine to patients with ESRD. Thirty-nine ESRD patients with hypertension were randomly assigned to receive losartan (n=13), enalapril (n=13), or amlodipine (n=13). Ultrasonic integrated backscatter (IBS) and serological markers of collagen type I synthesis and degradation were used to assess the degree of myocardial fibrosis just before and after 6 months of treatment. There were no significant differences in antihypertensive effects among the three agents. In the enalapril- and amlodipine-treated groups, the mean calibrated IBS values increased significantly after 6 months of treatment (enalapril: -31.6 +/- 1.3 to -29.4 +/- 1.2 dB, p=0.011; amlodipine: -30.6 +/- 1.4 to -27.2 +/- 1.2 dB, p=0.012). However, the mean calibrated IBS values in the losartan-treated group did not increase after 6 months of treatment (-31.2 +/- 1.7 to -31.3 +/- 1.4 dB, p=0.88). The ratio of the serum concentration of procollagen type I carboxy-terminal peptide to the serum concentration of collagen type I pyridinoline cross-linked carboxy-terminal telopeptide was significantly reduced in the losartan-treated group (42.6 +/- 4.6 to 34.4 +/- 3.6, p=0.038). The present study indicates that losartan more effectively suppresses myocardial fibrosis in patients with ESRD than does enalapril or amlodipine despite a comparable antihypertensive effect among the three drugs. PMID- 16471174 TI - Functional confirmation of Gitelman's syndrome mutations in Japanese. AB - Gitelman's syndrome is an autosomal recessive inherited renal tubular disorder resulting from loss-of-function mutations in the thiazide-sensitive sodium chloride cotransporter gene (SLC12A3). We have previously reported that the combined allele frequency for the reported Gitelman's syndrome mutations is 0.0321. However, almost all of the reported Gitelman's syndrome mutations were from case reports without functional confirmation. In the present study, we assessed the functionality of the two most prevalent mutations in Japanese, T180K and L849H, using a mammalian cell expression system. Human SLC12A3 cDNA was transiently expressed in Chinese hamster ovary (CHO) cells under the control of a cytomegalo virus (CMV) promoter. The T180K and L849H mutations were introduced by site-directed mutagenesis. The activity of the Na+-Cl- cotransporter was assessed by measuring tracer 22Na+ uptake. While the T180K variation was just a polymorphism, the L849H mutation was confirmed to be a loss-of-function mutation and appears to be responsible for the Gitelman's syndrome. This observation may have very important clinical implications, since the allele frequency of this variation is 0.0126. PMID- 16471175 TI - Class of antihypertensive drugs, blood pressure status, and risk of cardiovascular disease in hypertensive patients: a case-control study in Japan. AB - The purpose of this study was to compare the effect of different classes of antihypertensives on the risk of cardiovascular events in a case-control study of hypertensive patients. The subjects consisted of 171 hypertensive patients who had experienced a cardiovascular event and 537 randomly selected hypertensive controls who were matched to the cases by gender, age, and hospital/clinic. Both cases and controls had been under antihypertensive medication for at least 6 months before the onset of the cardiovascular event (cases) or before the enrollment (controls). A total of 134 physicians across the nation recruited cases and controls, and reported details of the prescription of antihypertensives and clinical and behavioral variables of their patients. Although there was no measurable difference in the risk of cardiovascular events according to the class of antihypertensives, statistically significant increases in the risk of cardiovascular events were observed for non-use of calcium antagonists among patients with angina pectoris and for non-use of the renin-angiotensin system inhibitor (angiotensin-converting enzyme inhibitor and angiotensin II receptor blockers combined) among patients with diabetes mellitus. Higher levels of blood pressure were associated with an increased risk of cardiovascular events. The findings suggest that appropriate control of blood pressure is more important in the treatment of hypertension than the choice of antihypertensives. PMID- 16471176 TI - Assessment of the microRNA system in salt-sensitive hypertension. AB - Most animal microRNAs are imperfectly complementary to their mRNA targets and inhibit protein synthesis through an unknown mechanism. MicroRNAs have been reported to play important roles in a number of biological processes. We assessed the microRNA system in Dahl salt-sensitive rats in order to investigate possible roles of microRNA in salt-sensitive hypertension. We constructed microRNA libraries from the kidneys of Dahl salt-sensitive and Lewis rats taking normal or high-salt diets (4 groups), and identified 91 previously reported and 12 new microRNAs expressed in the kidney. We then used Northern blotting to assess the expression levels of 118 microRNAs in the kidneys and heart ventricles. No significant differences in microRNA expression profiles were observed among the 4 groups. Thus, the microRNA system seemed to be unlikely to contribute to salt sensitive hypertension in Dahl salt-sensitive rats. PMID- 16471177 TI - Medullopressin: a new pressor activity from the renal medulla. AB - The kidney contains blood pressure-lowering substances, such as prostaglandin E2 or prostaglandin A2, and blood pressure-raising substances such as thromboxane A2. Most of the postulated substances, however, have not yet been isolated in a pure state and are of unknown structure. In the present study, we separated a chloroform extract from the medulla of pig kidneys using various chromatography procedures. Each fraction was tested in spontaneously hypertensive Wistar rats. One of these fractions caused a powerful blood pressure increase of 30.1 +/- 7.1 mmHg systolic and 34.7 +/- 6 mmHg diastolic (N=7; p=0.0003), reaching its maximum 55 +/- 27 s after completion of the injection and lasting for 201 +/- 59 s. A long-lasting contractile response in porcine and bovine coronary artery rings was observed. In the mouse aortic rings, the contractile response accounted for 0.38 +/- 0.13 g, ie., 31.9 +/- 10.9% of the maximum potassium response (N=11; p=0.003). Because this activity could not be attributed to any known vasoactive substance, it was considered to arise from a novel underlying active substance in the kidney medulla, which we named medullopressin. PMID- 16471178 TI - Chronic hypoxia accelerates the progression of atherosclerosis in apolipoprotein E-knockout mice. AB - The aim of this study was to investigate the effect of chronic hypoxia on the development and progression of atherosclerosis in apolipoprotein E-knockout (apoE KO) mice. Male and female apoE-KO mice (6 weeks old) and age- and sex-matched wild-type mice were kept under hypoxic conditions (10.0 +/- 0.5% O2) in a gas chamber or in room air for 3 weeks. Aortic atherosclerotic plaque was not observed in wild-type mice under normoxic or hypoxic conditions. In the apoE-KO mice, however, hypoxia induced proliferation of smooth muscle cells and plaque formation in the aorta, which were not observed under normoxic conditions. Although sexual dimorphism of the response to hypoxia was not observed, these hypoxia-induced atherogenic changes were accompanied by a significant increase of plasma low density lipoprotein (LDL) cholesterol and NADPH-dependent vascular superoxide (O2-) production. Furthermore, matrix metalloproteinase (MMP)-9 was activated in the aorta of apoE-KO mice. In conclusion, chronic hypoxia accelerated the development of atherosclerosis in apoE-KO mice, along with increased O2- production and activated MMP-9 in the aorta. PMID- 16471179 TI - A case of vertebral artery dissection associated with morning blood pressure surge. AB - We report a case of a middle-aged man who suffered a cerebral infarction resulting from dissection of a vertebral artery associated with morning blood pressure surge. A 56-year-old man was transferred to our hospital with dizziness and vomiting in the early morning on a cold day in winter. He reported that he had been standing in front of the sink after bathing when he suddenly felt dizzy and fell down. He did not lose consciousness, and by the time he reached the hospital by ambulance, his dizziness had subsided, but he complained of severe headache and vomited 3 times. On admission, he was alert, and there were no neurological or radiological abnormalities (CT, MR angiography) in the brain. However, infarction in the left cerebellar hemisphere was detected by brain MRI on the 5th day of hospitalization. String sign of the left vertebral artery was noted by angiography, confirming the diagnosis of dissection of the left vertebral artery. Ambulatory blood pressure monitoring was performed after discharge. Although the mean 24-h blood pressure was in the normal range, a marked morning blood pressure rise was observed. We speculated that the acute rise of blood pressure in the early morning might have contributed to the dissection of the vertebral artery. PMID- 16471180 TI - Renal artery dissection after angiographic evaluation of lower extremities. AB - We describe a case of renovascular hypertension that was caused by renal artery dissection after an angiographic evaluation of the lower extremities. Retention of contrast medium in the affected kidney even 1 day after the procedure caused us to suspect renal artery dissection. Magnetic resonance angiography revealed irregular streaks in the lumen of the affected artery, a sign of dissection. Treatment by a transluminal angioplasty with stent insertion improved renal function and reduced systemic blood pressure with normalization of plasma renin activity. In patients whose systemic blood pressure increases suddenly after any conceivable physical traction on the abdominal aorta, evaluation of the renal arteries should be considered. Appropriate endovascular treatment can preserve renal function and reduce systemic blood pressure. PMID- 16471181 TI - [Analysis of patients under 45 years of age with cancer of the larynx]. AB - INTRODUCTION: In last years we observe more frequently squamous cell carcinoma in patients below 45 years of age. Especially we examined patients with cancer of the larynx. MATERIAL AND METHODS: Estimation of surgical treatment of patients with cancer of the larynx under 45 years age after prior analysis of the following factors: localization of laryngeal carcinoma, local and regional advancement, the incidence of "occult metastases", the presence of metastases in the pre-laryngeal node, the regions of the neck affected by metastases. An analysis of a group of 140 patients who underwent surgery for laryngeal carcinoma in the period 1950-1998 was carried out. In all of the cases partial or total laryngectomy with unilateral or bilateral neck dissection was performed. RESULTS: In patients under 45 years of age, metastases to the lymph nodes were more frequently observed. The incidence of "occult metastases" and metastases to the pre-laryngeal node were more frequently. CONCLUSIONS: The cancer of the larynx in "young adults" is more aggressive and worse prognosis than in older patients population. PMID- 16471182 TI - [Concomitant radiochemotherapy followed by adjuvant chemotherapy in patients with poorly differentiated nasopharyngeal cancer; tolerance and early results of treatment]. AB - INTRODUCTION: Recently, concomitant radiochemotherapy became a method of choice in patients with poorly differentiated nasopharyngeal cancer. The aim of this study is to estimate tolerance and early results of the concomitant radiochemotherapy followed by adjuvant chemotherapy (modified US Head and Neck Intergroup protocol). METHODS AND MATERIAL: Analysing protocol consist of conventionally fractionated radiotherapy (TD = 70 Gy) given concomitantly with cisplatin (30 mg/m2 daily during 3 days every 3 weeks). This part of treatment was followed by 3 courses of PF (cisplatin + 5-fluorouracil) chemotherapy. Between August 1998 and September 2003 thirty six patients (27 male and 9 female) were qualified to treatment. Median age was 33 years. RESULTS: Tolerance of concomitant radiochemotherapy was acceptable. Intensive mucosal acute reactions (>G2) were observed in 67% patients. Life threatening complications (sepsis + DIC) was observed in single case. All patients received radiotherapy in planned total dose. Eighty six percent of patients received cisplatin in planned cumulated doses. Tolerance of the adjuvant chemotherapy was worse. Only 44% patients received all three courses of PF chemotherapy. The reasons of incomplete chemotherapy were neutropenia, infections, prolongated acute reactions or performance status decreasing. Complete regression was obtained in 86% patients. Two years overall and disease free survival rates were 83% and 72%, respectively. CONCLUSIONS: Our results confirm high activity of the concomitant radiochemotherapy followed by chemotherapy in patients with poorly differentiated nasopharyngeal cancer. Those results confirm also high toxicity of this regimen, what suggest very careful patients qualification to treatment. PMID- 16471183 TI - [Evaluation of diagnostic and rhinologic aspects in the treatment of lacrimal duct stenosis]. AB - INTRODUCTION: Diagnostic and rhinosurgical problems relevant to surgical treatment of lacrimal duct stenosis are discussed. MATERIAL AND METHODS: A group of 418 patients suffering from lacrimal duct stenosis were subjected at the same time to 455 procedures of endonasal dacryocystorhinostomy combined with other intranasal interventions, such as septoplasty, ethmoidectomy, mucotomy. Rhino ophtalmologic evaluation included: micro-endoscopic examination, CT of paranasal sinuses, dacryocystography or dacryoscintygraphy. RESULTS: In the most of the patients we noted at least some anatomical alterations of the nasal cavity. Most common were septal deviations (85.9%), followed by hypertrophy of the inferior turbinates (78,0%), pathology of ostio-meatal complex (24.8%), concha bullosa (15.0%) or various combinations of these. Chronic rhinosinusitis was noted among 23.2% of patients. Radiological examinations showed the predominance of postsaccal stenosis in 15.3-17.0% cases (respectively to the right or left side), saccal stenosis in 12.7-11.7% cases and presaccal in 4.3-3.6% cases. CONCLUSIONS: This combined procedure makes easy not only the operation on lacrimal duct but removes the pathologic changes and has a good influence on healing. PMID- 16471184 TI - [The sentinel node in cancer of the oral cavity, pharynx and larynx]. AB - AIM OF THE STUDY: The evaluation of the importance of the prognostic of sentinel node biopsy in head and neck cancer with N0 neck. MATERIAL AND METHODS: Prospective study included 19 patients with squamous cell carcinoma of the oral cavity, pharynx and larynx without enlarged lymph nodes in ultrasound examination (N0). The study was carried out in ENT Department of Medical University in Poznan between 2001-2004. Each patient underwent lymphoscintigraphy after administration of radionuclide Technetium 99m (Nanocol). In case of detection of sentinel node, blue dye was injected into the tumour bed and dyed lymph nodes were removed. The specimens were evaluated histologically and immunohistochemically. The patients are still followed up. RESULTS: So far 19 patients were included in our study. The tumours were classified from T1 to T4. In each case sentinel nodes were located. Metastases in removed lymphnodes were found in 5 patients. In one of them regional recidive was observed after 5 month. In the other patients no changes were found in follow up examination. In 14 cases in histological examination no metastases were found in sentinel nodes. In this group in one case local recurrence was observed and in another two lymph nodes metastases were confirmed. In patients with positive sentinel node elective neck dissection was performed. So far immunohistochemical examination has not provided us with any vital information. In no case with negative sentinel node we found micrometastases with the use of cytokeratin. CONCLUSIONS: Based on our preliminary observation of the patients we have found that there is certain correlation confirming the fact that sentinel node may be reliable for the whole neck lymphatic system. Further clinical observation are needed based on a larger number of patients and longer observation period. PMID- 16471185 TI - [Tuberculosis of larynx--still current problem]. AB - Article presents 4 cases of laryngeal tuberculosis hospitalized in ENT-department of Klodzko Hospital in years 2001-2003, being after-effect of advanced main process in lungs. Attention was turned onto shortcomings in Healt system reform after 1999, unfavourably influencing on early recognizing of new cases of tuberculosis. PMID- 16471186 TI - [Antioxidants enzymes activity and concentration of lipid peroxidation products in chronic tonsillitis before and after surgery]. AB - INTRODUCTION: Chronic tonsillitis is one of the most often disease in otolaryngological practice. Free radicals release during chronic inflammation process are responsible for tissue destruction. The aim of the study was to estimate the free radicals production and the activity of antioxidative enzymes during chronic tonsillitis before and after surgery. MATERIAL AND METHODS: The study was performed in group of 60 patients and 32 healthy persons as a control group. We analized the activity of enzymes in blood and serum from the patients before the operation and on 7 day, 4 week and 6 months after the operation. RESULTS: The obtained results shows a lower activity of superoxide dismutase before the operation. After the surgery the activity of the enzyme increased to values observed in control group. The catalase and glutathione peroxidase activity were higher during chronic inflammation, and after the operation the activity decreased, but it was still higher then in control group. Concentration of lipid peroxides products (malondialdehyde) in erythrocytes and serum were higher before the surgery. Then the concentration decreased to values observed in healthy subjects. CONCLUSIONS: The measuring and monitoring the activity of antioxidative enzymes and the concentration of lipid peroxidation products (malondialdehyde) could be useful in proper diagnosis of chronic tonsillitis and in adequately early qualification for surgery. PMID- 16471187 TI - [Treatment of patients with breathing disturbances during sleep in the II ENT Department Silesian Medical University Zabrze]. AB - INTRODUCTION: The treatment of snoring and obstructive sleep apnea syndrome comprises preventive behavior, conservative methods (CPAP) and surgical treatment. In the diagnostics procedures the most important aim is to determinate whether the patient is an habitual snorer or OSA patient. Very important thing is also patient's qualification for proper mode of treatment. The aim of the surgical treatment is to remove all the reasons that obturate the upper airways and cause the increasing of respiratory air stream resistance. The most collapsible airway region during sleep is oropharynx involves the soft palate, uvula, tonsils, tonsillar pillars, base of the tongue and pharyngeal muscles. MATERIAL AND METHODS: In the study the results of surgical treatment of 138 patients suffer from breathing disturbances during sleep treated at the ENT Department Silesian Medical University in Zabrze in the years 1997 to 2002 were presented. In the evaluated group were 51 habitual snorer and 87 OSA patients. Various type of surgical procedures were performed depend on the airway obturation level (septoplasty, tonsillectomy, LAUP, arytenoidectomy). The postoperative control sleep studies were performed in 68 patients (49.3%) 6 to 12 weeks after the end of the surgical treatment. In all the patients the subjective improvement were evaluated. RESULTS: In 35 from 45 OSA patients (77.8%) RDI decreased after the surgical treatment more then 50% of the preoperative value or was lower then 10 (RDI lower then 10 was in 16 patients, in 19 patients decreased more than 50%). The subjective radical improvement was obtained in 107 patients (77.5%), moderate improvement in 19 cases (13.8%) and no improvement in 12 patients (8.7%). CONCLUSIONS: On the base of our results we concluded that surgery is successful method for treatment of selected patients with breathing disturbances during sleep. PMID- 16471188 TI - [Velopharyngeal incompentance treated by the pharyngeal flap and the Orticochea pharyngoplasty--a comparison of speech results]. AB - INTRODUCTION: Velopharyngeal insufficiency is observed in about 10-40% of patients treated for cleft palate. Opinions on the effectiveness of applied methods differ in the assessment of specialists involved in the treatment of velopharyngeal insufficiency. MATERIAL AND METHODS: Since 2011, thirty-eight patients were treated for this defect at the Hospital of Polanica Zdroj. The report compares the effects of treatment by pharyngofixation and Orticochea's surgery. All the patients underwent prospective examinations--phoniatric evaluation with aural speech assessment, nasometry (using the device for objective and automatic nasality testing) as well as acoustic speech analysis (by means of MDVP). RESULTS: The effectiveness of both modalities of treatment was evidenced on the basis of statistical analysis of the findings of the perceptual speech evaluation as well as differences-- p > 1. Moreover, the statistical analysis of nasometric examinations revealed improvement achieved after the use of both methods, in this case Orticochea's method being considered slightly more effective-- p = 0.084. CONCLUSIONS: Concluding, both operations may be considered safe and effective in the treatment of velopharyngeal insufficiency in cleft palate patients. PMID- 16471189 TI - [Nose reconstruction in material of Maxillofacial Surgery Department at the Pomeranian Academy of Medicine in Szczecin in 1975-2004]. AB - INTRODUCTION: The focus of this paper is the evaluation of usability of various nose skin reconstruction methods applied after removal of a skin cancer in the Teaching Hospital of Maxillofacial Surgery at the Pomeranian Academy of Medicine in Szczecin in 1975-2004. MATERIAL AND METHODS: The research material consisted of 285 patients who underwent a surgical treatment of nose skin cancers located in various skin areas. In 67 cases, the tumour was situated on a nostril flare, in 42--on the nose tip, in 98--on the bridge and in 68--on one of its sides, most frequently, near the corner of the eye. Said tumours often covered more than one of the above-mentioned areas or even all of them. Reconstruction planning and performance was an integral part of the surgical treatment. The tumour amputation was preceded by an assessment of the quantity and quality of tissues necessary for filling in the lesions. Lesions located on the nose surface were covered by free skin grafts. In the case of nostril flare tumours, they were filled in with grafts composed of skin and cartilaginous tissue from the ear. Lesions including all nose layers were most frequently (in 136 cases) covered with various types of axially vascularised frontal skin flaps i.e. by the middle or temporal part of the frontal flap. In the case of elderly people the nasolabial flaps were used. RESULTS: In most cases, both the aesthetic and functional effects were very good. What appeared to be the most difficult reconstruction case was the restoration of the nasal bone structures after their complete amputation. CONCLUSION: Due to the thickness, colour and easiness of preparation of the middle frontal flap, it is considered to be the best material for reconstruction of the nose. PMID- 16471190 TI - [Multiple symmetric lipomatosis in the otolaryngology as diagnostic and therapeutic problem]. AB - Multiple symmetric lipomatosis (MSL) is a systemic disease connected with a degeneration of the adipose tissue. Association of reduced glucose tolerance, hyperinsulinemia, hyperlipoproteinemia, hyperuricemia, macrocytic anemia and renal tubular acidosis, polyneuropathy have been described. Lipomatosis was initially described in 1846 by Brodie but the exact aetiopathogenesis is still unknown. Depending on the anatomical location of the lipomatosis we can divided MSL into two types. MATERIAL, METHODS AND THERAPY: We describe the findings in three patients with symmetric lipomatosis: two (n=2) with the first and one (n=1) with the second type of the disease. The patients were diagnosed and treated in the Department of the Otolaryngology during last of two years (2002 to 2004). Patients had executing following research: ultrasonography of the neck and abdominal cavity (n=2), radiological examination of the chest (n=3), computer tomography of the neck (n=1), thin-needle biopsy (n=3), histopathological examination (n=2) and laboratory investigations (n=3). Two patients received the pharmacological treatment (magnesium and the vitamin B6) and we observed marked regress of the disease. In the patient who at first did not agree for the treatment, came up to the heavy increase mass of tumors, especially on the neck. Liposuction and the pharmacological treatment were executed in this patient. He is still in the observation. RESULTS: In case of tumors of face and neck, we have to take into account degenerative processes of the adipose tissue. Our observations indicate the efficiency of the magnesium and the vitamin B6 therapy in patients with multiple symmetric lipomatosis. The obtainment in this range of reliable conclusions require of research at the greater number of patients. We want to underline that there are any research of the influence of the magnesium and the vitamin B6 on the course of this disease in the literature. PMID- 16471191 TI - [Treatment of lymphangioma with OK432 in children]. AB - From 1998 to 2003 6 children with lymphangioma of cervico-facial region were treated with OK-432 in Polish Mother Health Institute. Three of them had nonresectable leasion and two required tracheostomy during first months of life. Each patient received from 1 to 10 sessions of obliteration. In total 23 sessions were performed. The youngest patient treated with OK-432 was 2 weeks and the oldest 2 years old. The response to treatment was very good in 4, good in 1 and fairly good in 1 case. In three patients criodestruction was applied as an additional treatment. In two children residual lesions were subsequently removed surgically. There were no serious complications. After obliteration sessions only 2-3 day temperature increase and local inflammation occurred. We consider obliteration with OK-432 as a treatment of choice in lymphangiomas in children. PMID- 16471192 TI - [Otoacoustic emissions and recruitment]. AB - INTRODUCTION: Typical high frequency hearing loss is accompanied by an exaggerated response to loudness as the sound level exceeds your elevated thresholds. This exaggerated response to sound levels is termed "recruitment". Recruitment is a paradoxical phenomenon present in hearing losses due to cochlear damage. Loudness recruitment is usually present to some degree as measured on the SISI. There have been many explanation connected this phenomena so far. It has been suggested that otoacoustic emissions might be more sensitive in assessment of changes to the cochlea caused by noise than pure tone audiometry. MATERIAL AND METHODS: In the present study results of the short increment sensitivity index test (SISI), DPOAE as well as slope of DPOAE input/output (I/O) function were compared before and after noise exposure. The I/O function plots DPOAE amplitude as a function of the level of the primary tones, for the progressively increasing stimulus levels (from 45 to 70 dB SPL) for the frequencies 1, 2, 3, 4, 6, 8 kHz. From 40 patients between 18-21 years, 30 ears were extracted with cochlear lesions and good hearing. RESULTS: Both, PTA and DPOAEs amplitude showed in this group significant reduction due to noise exposure. Slopes of the DPOAE I/O function were depended on stimulus levels. For the low intensity stimuli levels (45-55 dB SPL) we obtained DP I/O amplitude value worse than before noise exposure. When stimulus level was above 55 dB SPL for the frequencies (1, 2, 3, 4, 6, 8 kHz) the amplitude and DP I/O slope was higher than before noise exposure. We did not notice that phenomena in persons without cochlear lesions. CONCLUSIONS: Our results showed that in examination group with cochlear lesion I/O DP amplitude was higher using stimulus level above 55 dB SPL. This notice can be otoacoustic recruitment phenomena. PMID- 16471193 TI - [Importance and usefulness of laryngological screening of the preschool children population in conductive hypoacusis diagnostics]. AB - INTRODUCTION: Incidence of upper respiratory tract ailments among children between 2 and 9 years old is very common. Discreet symptoms, like low grade conductive hypoaccusis or articulation's disorders are ignore or imperceptible by parents. The survive of these disorders in the child's development stage has a direct influence on cognitive functions, speech and social development and progress in learning. The aim of the work was to prove the efficiency of laryngological screening among asymptomatic population of preschool children on the basis of comparison of two difference populations: for the first time examined (Szczecinek) and at regular intervals of one year (Wolsztyn). MATERIAL AND METHODS: In the town Szczecinek and in the town Wolsztyn laryngological screening among 1172 (762 and 410 respectively) children was carried out. All the participating individuals were fully laryngological examined and tested with tympanometric audiometry. RESULTS: On the base of these examinations in the town Wolsztyn the following was to state: 46 (11.2%) children were refered to surgical treatment. In this group in 31 (67.4%) pathological tympanometric curve b- and c- type were affirmed. 42 (10.2%) children were refered to conservative treatment. In the town Szczecinek 142 (18.6%) children were refered to surgical treatment. In this group in 81 (57%) pathological tympanometric curve b- and c- type were affirmed. 110 (14.4%) children were refered to conservative treatment. CONCLUSIONS: (1) Usefulness of laryngological screening was confirm of the proportion of laryngological findings in the population of healthy seeming children refered to treatment. (2) The screening role is higher at rural area by low acsses to ENT consultants. Every year repeated screening influence population health, conscious approaching the ENT healthcare. PMID- 16471194 TI - [Bilateral neck tumor as the clinical manifestation of neurofibromatosis type I (NF I)]. AB - A case of the 17-years old girl with bilateral tumors of the neck due to neurofibromatosis type I has been described. Patient has been unsuccessfully treated conservatively for over two years prior to the admission to our department. The principles of diagnostics and surgical treatment of the patients with neurofibromatosis type I have been presented. PMID- 16471195 TI - [Tympanoplasty decision making in a child with HIV contamination--case report]. AB - Otologic problems are infrequent in HIV patients. Authors present the case of chronic otitis media in the HIV infected child. The patient with significant hearing loss, and prolonged aural discharge, resistant to local and systemic antibiotic treatment was treated surgically. Mastoidectomy with myringoplasty was performed. The substantial hearing improvement has been obtained, ear discharge never returned. However due to episodes of chronic otitis media with effusion ear drainage with tympanostomy tubes, after initial surgery have been performed two times. The controversies regarding surgery decision-making in this group of patients are discussed. PMID- 16471196 TI - [Fatal necrotizing mediastinitis as a complication of peritonsillar abscess]. AB - In the paper showed the case of fatal necrotizing phlegmon of the neck and descending mediastinitis in 48 year old male patient with primary peritonsillar abscess. On the base of literature it was showed the etiology, pathomechanism of the evolution of this heavy complication and surgical and pharmacological methods of its treatment. In spite of prompt wide incision and drainage of phlegmon of the neck and mediastinum and intensive treatment in intensive care unit conditions patient died in the image of septic shock and pus changes in the mediastinum, heart and lungs, what was showed on the base of post-mortum examination. PMID- 16471197 TI - [Rare case of pyogenic granuloma of the tongue in an 8-week-old child]. AB - In paper a case of pyogenic granuloma of the tongue in an 8-week-old child was presented. The signs and symptoms, etiological agents, surgical treatment, and complications are discussed. This case is the youngest case with an unusual location. The lesion was presented as sessile, soft, single and pink erythematous nodule with tendency to spontaneous hemorrhage, 3 cm in diameter. We performed conservative excision because of anaemia and not good condition of the child. We didn't observe recurrence of this lesion in follow-up after 2 weeks, 6 months and 3 years. We report this case to alert the otorhinolaryngologist of this rare condition. There have been very few reports in the literature of this condition. PMID- 16471198 TI - [Rhabdomyosarcoma of nasopharynx and oropharynx in adult--a case report]. AB - The authors present the case of rhabdomyosarcoma--a rare malignancy--in the adult patient. The epidemiology, etiopathogenesis, diagnostics and therapeutical options were discussed. PMID- 16471199 TI - [Haemangiopericytoma of the parotid gland]. AB - Haemangiopericytoma is a vascular, soft tissue neoplasm with rare occurrence in the head and neck region. On the basis of literature we discuss etiology, symptomatology, histopathology, diagnosis, treatment and prognosis. We present the case of 66-old woman with haemangiopericytoma in the parotid gland treated by surgery. PMID- 16471200 TI - [Nasal glioma--case report]. AB - Benign congenital lesions resulting from the deficient regression of neuroglial tissue in normal embryonic development are called gliomas. They are usually located in the nasofrontal region and are diagnosed in the postnatal period. They are included in the differential diagnosis of nasofrontal midline masses. We present a case of the 9-year-old boy treated in the ENT Department of Wroclaw Medical University with the initial diagnosis of nasal polyposis or tumour which appeared to be glioma in the postoperative histological evaluation. The tumour was removed endoscopically. We present magnetic resonance imaging of the case together with the review of the literature concerning gliomas. We conclude that each doctor has to be aware of possible neoplasm even in the youngest groups of the patients. PMID- 16471201 TI - [History of treatment and classification of maxillary sinus neoplasms]. AB - History of treatment of maxillary malignant neoplasms goes back to the middle of 19th century, when Gensoul and Lizzard performed their first maxillectomy. However the development of maxillar surgery is connected with achivements of Moure, who as the first one characterized and applied innovative skin cut on the face. Author presents the history of other surgical procedures in therapy of maxillary tumors and describes methods which are apply nowaday. Attention is paid to the Polish contributions and accomplishments in diagnostics and treatment of tumors localized in this area. The classification of maxillary tumors from Sebileau dissertation till the newest TNM classification from 2001 is described. PMID- 16471202 TI - [The hypertrophy of pharyngeal tonsill in the estimation of XIX-century physicians]. AB - The role of such physicians as Rudolph A. Kolliker, Johann N. Czermak, Ludwig Tuerck, Friedrich E. Voltlini, Benjamin B. Loewenberg, Hubert v. Luschka in the building of the science on hypertrophy of pharyngeal tonsil are pointed out first of all. The specific significance of achievements of Wilhelm Meyer (1824-1895) of Copenhagen based on scrupulous examinations is described. In the Polish territories the innovatory were works of Teodor Heryng, Klemens Koehler, Wladyslaw Wroblewski, Jan Sedziak, Teofil Zalewski. The epidemiology, etiology, pathogenesis, anatomo-patological examinations, the role of pharyngeal tonsil on immunity system are presented widely. The influence of hypertrophy of pharyngeal tonsil on general evolution of child, on his intellectual sphere, on speech disorders, on upper respiratory tract disorders, on auditory disorders, on the origination of deaf-mutism, on the origination of reflexogenous neuroses are described in more detail. An important role of anterior rhinoscopy, posterior rhinoscopy, and digital examination in the diagnosis of this illnes are also presented with full particulars. PMID- 16471203 TI - [Report from the 10th International Congress on Surgical and Prosthetic Rehabilitation after Laryngectomy 17-20.04.2005 Groningen, Holland]. PMID- 16471204 TI - [Highly virulent Streptococcus suis infection and problems involved in the disease prevention and control in China]. PMID- 16471205 TI - [Anlysis on features of dead cases with human Streptococcus suis infections]. AB - OBJECTIVE: To describe the clinical and epidemiological features of dead cases with human Streptococcus suis infections, and to find the target population for preventing death and the related indicators. METHODS: Epidemiological investigation on human Streptococcus suis infections was implemented used unified questionnaires. Analysis on dead cases and survival cases (as contrast) was done. RESULTS: The population with highest fatality rate was in 40-49 age group. 97.37% of dead cases had toxic shock syndrome. The mean interval from onset to admission was 0.76 days, and the mean course was 2.11 days. The progression among dead cases was faster than that among survival cases. Chief clinical manifestations of dead cases that are more frequent than survival cases are purpura (73.68%), diarrhea (50.0%), dyspnea (21.05%), conjunctival congestion (34.21%), etc. Renal impairment and liver involvement in dead cases were more significant than that in survival cases. No significant difference between mean incubation period, exposure rates of main risk factors in dead cases and in survival cases was found. CONCLUSION: Preventing toxic shock syndrome might reduce the fatality rate. The target population for preventing death is aged > or = 40. Liver function and renal function testing might be indicators for monitoring the progression of human Streptococcus suis infections. PMID- 16471206 TI - [Matched case-control study for risk factors of human Streptococcus suis infection in Sichuan Province, China]. AB - OBJECTIVE: To study the potential risk factors of human infecting with Streptococcus suis. METHODS: 1: M matched case-control study was conducted. 29 human cases of Streptococcus suis infection in the early phase were included in the case group, Patients' family members, neighbors and peoples who had worked together with patients to handle deceased or sick pigs in the last week were recruited as matched controls. There were 147 controls in total. Both cases and controls received questionnaire investigation including the ways to contact sick/dead pigs. Conditional logistic regression was employed to analyze matching data. RESULTS: According to the results of multivariate analysis, slaughtering (OR = 11.978, 95% CI: 3.355-42.756), carcasses cutting and processing (OR = 3.008, 95% CI: 1.022-8.849) sick/dead pigs were associated with cases related to human Streptococcus suis infection. The attributable risk proportion were 91.65% and 66.76% respectively. The other types of exposures to sick/ dead pigs, including feeding, selling, burying and eating, were not associated with the human Streptococcus suis infection in our study population. CONCLUSION: Slaughtering, carcasses cutting and processing sick/dead pigs were important risky behavior for humans to be infected by Streptococcus suis. PMID- 16471207 TI - [Detection of virulence-associated factors of Streptococcus suis by multiplex PCR assay]. AB - OBJECTIVE: To rapidly and sensitively detect the four virulence-associated factors of Streptococcus suis, a multiplex PCR was developed. METHODS: In the process of this reaction, four distinct DNA targets were amplified. One target was based on the serotype 2 (and 1/2) specific cps gene and the others were based on Streptococcus suis mrp, epf (epf*) and sly gene, encoding the MRP, EF(EF*) and Sly proteins of Streptococcus suis. 72 isolates, which including 48 strains of Streptococcus suis and 24 strains of negative control, and 49 clinical specimens were detected by the multiplex PCR assay. RESULTS: All PCR products were detected by electrophoresis on 1.2% agarose gels. With the 48 Streptococcus suis strains, the positive detection rates of cps2+, mrp+, epf+, epf*+ and sly+ were 16/48, 14/48, 12/48, 3/48 and 26/48,respectively. The results were confirmed by bacteriological examination. There were no specific amplification products including 49 clinical specimens and 24 negative control strains. CONCLUSION: The results demonstrated that multiplex PCR was a highly specific and sensitive diagnostic tool for the detection of virulence-associated factors of streptococcus suis. PMID- 16471208 TI - [Human infection due to Streptococcus suis]. PMID- 16471209 TI - [The status of lead poisoning and the revelant research in children in China]. PMID- 16471210 TI - [Study on blood lead level and related risk factors among children aged 0-6 years in 15 cities in China]. AB - OBJECTIVE: To observe blood lead level and related risk factors among children aged 0-6 years old living in cities in China and to provide data for policy development to the prevention on environmental lead pollution. METHODS: A stratified-clustered-random sampling method was used. 17 141 peripheral blood samples of 0-6 years old children from 15 cities in China were tested. Tungsten atomizer absorption spectrophotometer was employed to determined the blood lead level. Related factors were also studied using a standardized questionnaire. Data were analyzed on related risk factors that affecting blood lead levels through multiple regression method. RESULTS: The mean blood lead level of 0-6 years old children from 15 cities in China was 59.52 microg/L including 10.45% of those > or =100 microg/L, and 0.62% > or = 200 microg/L. However, the blood lead levels seemed to have had a trend of increase parallel to age among 0-6 years-old children and were higher for boys (59.50 microg/L) than girls (54.95 microg/L). The risk factors which influenceing children's blood lead levels would include the type of housing, parent's education levels, social status and hobby, children's behavior habit, dieting habit and nutritional condition. CONCLUSION: The blood lead levels of children in China were lower than data gathered from former national studies but higher than those from developed countries, suggesting that the. Government and the whole society should be aware of the problem on lead poisoning among children during their childhood. PMID- 16471211 TI - [A study on the sero-epidemiology of hepatitis B in Chinese population aged over 3-years old]. AB - OBJECTIVE: To better understand and measure the impact from immunization activities over the last 10-years, we conducted a sero-epidemiological study using the remaining blood samples of a nationwide survey on Nutrition and Health in Chinese residents, in 2002. METHODS: HBsAg, anti-HBs and anti-HBc in blood samples were tested using enzyme-linked immunosorbent assay(ELISA) reagents and revised by solid phase radioimmunoassay and Abbott ELISA reagents. RESULTS: Among population older than 3 years, the prevalence rates of HBsAg, anti-HBs and hepatitis B virus(HBV) infection were 9.09%, 37.48% and 50.04% but for children between 3 and 12 year olds, they were 5.03%, 45.33% and 29.10% respectively. Among population older than 3 years, the rates of HBsAg and HBV infection in urban area were 4.61% and 43.51%, but the rates in rural area were 9.41% and 56.77% respectively while for children from 3-years to 12-years old, they rates were 2.10%, 20.45% in urban area and 8.25%, 39.22% in rural areas, respectively. CONCLUSION: Results revealed that since the hepatitis B vaccine was introduced into the expanded program on immunization(EPI) from 1992, the prevalence rates of HBsAg and HBV infection, especially in the children from 3 to 12 years of age, had obviously declined, suggesting that some changes had happened in the epidemic characters of hepatitis B in China. PMID- 16471212 TI - [A case-control study on the association between genetic polymorphisms of metabolic enzymes and the risk of colorectal cancer]. AB - OBJECTIVE: To investigate the association between metabolic enzymes polymorphisms and the risk of colorectal cancer(CRC). METHODS: Methods of detection used were based on polymerase chain reaction(PCR) including PCR-restriction fragment length polymorphism (PCR-RFLP), allele specific-PCR (AS-PCR) and multiple-PCR to identify the polymorphisms of CYP1A1 6235T/C, CYP1A2 734C/A, CYP2E1 -1259G/C, CYP2E1 -1019C/T, GSTM1 and T1 null type, NAT1 and NAT2 alleles among 140 cases and 343 cancer-free controls. RESULTS: The allele frequencies of CYP1A1 6235C, CYP1A2 734A, CYP2E1 -1259C, CYP2E1 -1019T, GSTM1 and T1 null type, NAT1* 10 and NAT2 Mx (x = 1,2,3) alleles were 31.65%, 63.77%, 23.02%, 32.61%, 57.25%, 17.39%, 26.45% and 39.21% in the case group and 39.85%, 66.62%, 20.27%, 28.61%, 55.46%, 20.35%, 25.22% and 39.36% in control group, respectively. The frequencies were in Hardy-Weinberg equilibrium. Data on single genetic polymorphism and stratification analysis of multi-genetic polymorphisms indicated that CYP1A1 6235CC homozygote was associated with the significant reduction of CRC risk (OR = 0.79, 95% CI: 0.63-0.99) and in individuals with CYP1A2 734A allele. CYP1A1 62345C allele had the same effect (OR = 0.53, 95% CI: 0.34-0.83). However, individuals with GSTT1 null genotype, GSTM1 null genotype could significantly increase the risk (OR = 4.41, 95% CI: 1.21-16.10). CONCLUSION: CYP1A1 6235C allele might play an important role in fighting against colorectal carcinogenesis. However, GSTM1 and T1 null genotype might serve as risk factors genetically. Larger scale population-based studies were needed to confirm the current findings. PMID- 16471213 TI - [A parental case control study on the association between reduced folate carrier gene polymorphism and neural tube defects]. AB - OBJECTIVE: To study the association between reduced folate carrier gene (RFC1 A80G) polymorphism and the risk for child with neural tube defects (NTDs), and to provide epidemiological evidence for the existence of NTDs genetic marker. METHODS: RFC1 (A80G) genotypes were detected using RFLP-PCR for blood DNA of 104 families with NTDs-affected children and 100 control families with no history of child-affected birth defects. Case-control study and transmission/disequilibrium test(TDT) for the RFC1 genotype of NTDs pedigree were carried out. RESULTS: The G allele frequency of children with NTDs was higher than that of controls when compared to A allele( OR = 1. 64, 95% CI :1.08-2.49). The offspring of the GG genotype were associated with a 2.56-fold increased risk of NTDs when compared to the AA genotype (OR = 2.56, 95% CI: 1.04-6.36) in our study population. There was evidence of association between G allele and the risk of parent having a child with NTDs (OR = 1.56, 95% CI: 1.07-2.28) in the TDT analysis. CONCLUSION: Our findings indicated that there was potential association between offspring RFC1 GG genotype and the risk of NTDs, and the G allele was a possible susceptible gene marker for an increased NTDs risk in the Chinese population. PMID- 16471214 TI - [Study on direct economic-burden and its risk factors of osteoporotic hip fracture]. AB - OBJECTIVE: To investigate the net "per case" direct economic burden of osteoporotic hip fracture of in-patients at Peking Union Medical College Hospital (PUMCH), and to explore the related factors which will influence the burden of the disease. METHODS: Data was from a retrospective questionnaire on 90 osteoporotic hip fracture in-patients of PUMCH above 50 years old from 1998 to 2002. The direct cost of fracture consisted of inpatient care, outpatient care, home care, appliance, and nutrition etc. Multiple statistics method was used to analyze the factors affecting the direct costs of disease. RESULTS: The average age of the studied population was 71.78 +/- 9.7 years, including 29 males and 61 females. Yearly direct economic burden for one patient of osteoporotic hip fracture was 32 776 RMB, in which hospitalization expense was 23 107 RMB, accounting for 70.5%. Factors which influencing total direct costs would include: length of hospitalization, occupation and the outcome of disease, while types of hip fracture,occupation and the length of hospitalization affecting the cost during hospitalization. The outcome of disease, length of hospitalization and rehabilitation influenced the cost during convalescence while age, type of payment and treatment, did not show significant influence on the cost of hip fracture. CONCLUSION: Osteoporosis had become a serious public health problem among the elderly, bringing about the increase of economic burden all over the world both on social healthcare and families of the patients. Our data provided base for preventive health care or health insurance on osteoporosis. PMID- 16471215 TI - [A case-control study on risk factor of Kaposi's sarcoma in Xinjiang]. AB - OBJECTIVE: To analyse related risk factors of classic Kaposi' s sarcoma in Xinjiang. METHODS: A 1:4 case-control study was used and the conditional logistic regression model was performed in this study. Cases were followed up in Xinjiang while controls were selected by the same sex, nation and age within 5 years with cases. RESULTS: Interleukin-6,vascular endothelial growth factor, beta-MG, neopterin, human herpevirus 8, were found to be associated with Kaposi's sarcoma risk in one-way variance model while beta2 -MG and human herpevirus 8 entered the multiple conditional logistic regression model, and their ORs were 1.002(95%CI: 1.000-1.003), 81.041 (95%CI: 1.790-3669.620). CONCLUSION: There was a correlate relationship between beta2 -MG and classic Kaposi's sarcoma being found that human herpevirus 8 exposure related factors seemed to have played important roles on classic Kaposi's sarcoma in Xinjiang. PMID- 16471216 TI - [A case-control study on the risk factors related to obstetric brachial plexus palsy]. AB - OBJECTIVE: To identify the risk factors and related degrees associated to obstetric brachial plexus palsy(OBPP). METHODS: A case-control study was performed. Neonatal records of thirty-one cases with OBPP and their corresponding maternal records from the Department of Gynecology and Obstetrics of eight hospitals in Shanghai city from 1988 to 2002 were reviewed. Four controls, all living in Shanghai were selected to match each case and were born within the same year at the same hospital. The control group also included 124 cases without OBPP. According to the uniformed data and tables used were from medical records and from pregnant women. Epidemiological study was carried out on both case group and control group. Variables for analyses would include: (1) race, age, height, family history, pre-pregnancy weight, body mass index at the pre-pregnancy (weight/height2) on those pregnant women as well as on parity of their mothers; (2) the process of delivery which includeing clinic pelvis evaluation, height of uterus, abdomen circumference,antepartum weight,body mass index before delivery, mode of delivery,the duration of active phase and 2nd stage of labor, shoulder dystocia; (3) on neonates: sex, gestational age, birth weight,affected limb, Apgar scores of 1 and 5 minutes, other birth trauma and resuscitation of infant. Statistical tests applied to these data would include Student's T test for continuous variables and chi2 analysis for discrete data. Risk calculation of OBPP was performed by univariable and multivariable conditional logistic regression analysis. RESULTS: 12 factors related to expsure were identified for OBPP through univariable conditional logistic regression analysis. When multivariable conditional logistic regression model at P = 0.1 was applied, four factors such as cesarean (OR = 0.060), forceps (OR = 65.237), birth weight (OR = 35.468), and pre-pregnancy body mass index (OR = 23.901) were selected. CONCLUSION: Forceps delivery,macrosomia, and increase of pre-pregnancy body mass index (> or = 21) were risk factors of OBPP in the order of degrees to risk while cesarean seemed to serve as a protective factor. PMID- 16471217 TI - [Study on the identification of HIV/AIDS and its surveillance system in rural areas of central China]. AB - OBJECTIVE: To explore the effective strategies for human immunodeficiency virus (HIV) surveillance and identification in rural areas of central China. METHODS: In a selected rural prefecture area of central China, an epidemiological investigation was conducted for all reported HIV/AIDS cases. A historical and analytic review was performed, with particular interests in examining the HIV epidemic reporting and identification system. RESULTS: Among all 626 reported HIV infected individuals in the selected prefecture, 86.1% (539/626) of them were infected through commercial plasma donation or clinically operated blood transfusion. With respect to disease surveillance and identification in the area, 52.2% (327/626) of all the cases were reported by hospitals or clinics. The number and proportion of HIV/AIDS cases identified or reported at county, prefecture, and provincial levels were 207 (33.1%), 303 (48.4%) and 116 (18.5%), respectively. The number and proportion of HIV/AIDS identified through specific epidemiological investigations and/or voluntary testing had been increased in recent years. In addition, among HIV/AIDS cases that were clinically identified, the proportion of those who were identified as outpatients had steadily increased,with the highest proportion (59.3%) observed in 2004. CONCLUSIONS: In rural areas of central China where the major mode of HIV transmission was through commercial plasma donation or clinically operated blood transfusion, hospitals and clinics seemed to have played and would continue to play important roles regarding HIV identification and surveillance. The role of institutions or settings at the prefecture level regarding HIV identification and surveillance should not be ignored. PMID- 16471218 TI - [Study on an epidemic of human lung plaque in Nangqian county, Qinghai province]. AB - OBJECTIVE: To study an epidemic of human lung plague fulminant from September to October, 2004 in Nangqian county, Qinghai province. METHODS: Cases were diagnosed through data from epidemiological, clinical, bacteriological, serological and autopsy studies. RESULTS: 14 patients were identified, ending up with 6 deaths and 8 cured. The first case was diagnosed as primary pesticemia late progressed to lung plague. 4 cases were transformed from pesticemia out of 13, leaving the 9 cases as primary lung plague. Situation was under complete control through routinely handling the plague focus. CONCLUSION: The first case was bitten by the infected fleas which parasitized the marmota preyed on a dog but later these fleas were brought into the tent by the dog. The others cases were infected through droplets or dust. Programs on monitoring and controling the amount of marmotas and fleas should to be strengthened to prevent the epidemics of plague in the area. PMID- 16471219 TI - [Seroepidemiology study on Helicobacter pylori (cagA, vacA) infection of elderly people in Beijing]. AB - OBJECTIVE: To assess the prevalence rate of Helicobacter pylori (H. pylori) in blood serum, its affecting factors and isoforms (CagA,VacA )infection in the elderly people in Beijing. METHODS: 2006 residents were investigated through household questionnaire in different areas of Beijing (urban, suburban and mountainous district), who were older than 60 years old. Serum H. pylori CagA, VacA and Ureas antibody was detected by immunoblotting. RESULTS: The total H. pylori infection rate was 83.4% and the infectious rate of I form pathogenic H. pylori was 56.0%. The incidence rate in urban or suburban district was higher than that of in mountainous district (P < 0.001). I form H. pylori infection rate in people with heavy labor or young elderly were higher than that of intelegent work or older elderly (P < 0.05 ). I form H. pylori infection rate in people of low diet was higher than that of high protein diet (P < 0.001). CONCLUSION: The rate of H. pylori infection in blood serum was high among the elderly people in Beijing with most of it belonged to type I . However, significant differences were noticed on the distribution of isoforms in different age groups, areas, professions and diet habit. PMID- 16471220 TI - [Study on a monitoring program regarding leptospirosis in some fore-and-after flood-affected along large rivers in Anhui province]. AB - OBJECTIVE: The study was designed to find out the epidemic characteristics of leptospriosis and to develop effective intervention measures. The effects of floods on leptospriosis in some areas along Yangzi river and Huai river in Anhui province was also analysed. METHODS: Study on serum epidemiology of leptospriosis was carried out from serous samples collected from native residents and animal hosts including isolation of pathogens at different phases (before,middle and after) and different monitoring spots,during the floods. RESULTS: Infection rate with leptospriosis pathogen among native residents was 13.49% during the flood period,much higher than 2.18% at post-flood (chi2 = 22.78, P < 0.01) stage, in the flood-affected areas along Yangzi river in 1998. The average rates of infection were 2.48% and 5.35% in affected and unaffected areas along Huai river respectively, in 2003. CONCLUSIONS: There was full evidence that floods causing the epidemics of leptospriosis. However, the transmission of leptospriosis among people would depend on affecting factors as scales of floods, lasting time, coincidence between flood happening and epidemic season, immuno-protection level against leptospriosis among people and so on to a great extent. Factors as the magnitude of pathogens carried by various kinds of infectious sources were also important determinants affecting the nature, being epidemic or pandemic of leptospriosis. It was suggested that active surveillance network on the sources of infection and risk factors of leptospriosis should be developed for the control and prevention of the disease, in the flood-hit areas. PMID- 16471221 TI - [Study on the timeliness of the notifiable communicable diseases surveillance system in Fujian province, China, 2004]. AB - OBJECTIVE: To understand the timeliness of the notifiable communicable diseases surveillance system in Fujian province. METHODS: Database from the internet based communicable diseases reporting system was used. RESULTS: The 50th percentile of time between the disease diagnosed and report recorded in medical faculties was 1 day in 2004 which was 6 days less than that in 2001 - 2003. The timeliness rate of 0 day was 46.46%, a 2.7 times over that in 2001 - 2003. The timeliness of notifiable communicable diseases surveillance system in different administrative areas, reporting units and on different diseases was significantly different. Time between the disease diagnosed and report recorded was the shortest in those cases reported by hospitals and traditional Chinese medicine(TCM) hospitals at the county level and above, with 50th percentile as 0 day, but the timeliness rate of 0 day was 50.76% with 70.04% of the cases were reported from hospitals and TCM hospitals of county level and above. Length between the disease diagnosed and reported was the longest in those cases recorded by Centers for Disease Control and Prevention(CDCs) with the 50th percentile as 3 days. The source of cases recorded by CDCs came from hospitals at the township level, where there was no connection to internet but the reporting cards had to be sent to local CDCs. Time between the disease being diagnosed and reported was 2 days in those cases reported by hospitals at the township level. 21.21% of cases were recorded by hospitals of township level and CDCs. The 50th percentile of time shown between the reported records and confirmed by CDCs was 4 hours The 24 hour timeliness rate was 63.65%. CONCLUSION: The timeliness of the notifiable communicable diseases surveillance system had been improved significantly after the medical personnel recording the cases directly through internet. Timeliness could be further improved through access to internet at the hospitals of township level, training of staff and better hospital management systems. PMID- 16471222 TI - [The analysis on single nucleotide polymorphism of PDCD1 and systemic lupus erythematous susceptibility among Han indigenous in Chinese population]. AB - OBJECTIVE: To explore the frequencies and polymorphic distribution of intron 4 of programmed cell death 1 (PDCD1) gene, and to analyze its relation to the susceptibility of developing systemic lupus erythematosus (SLE) in indigenous Han Chinese from Anhui province. METHODS: Blood samples from 122 patients with confirmed SLE and 143 controls were collected for a case-control study. DNA of the subjects was extracted and amplified by polymerase chain reaction (PCR). The single nucleotide polymorphisms(SNPs) of PDCD1 7809 locus,7872 locus and 8162 locus were further confirmed by direct DNA sequencing and BLAST. RESULTS: The PDCD1 7809 locus was indentified as type G/G among all the subjects investigated. There were significant difference at PDCD1 7872 locus with C/T polymorphism and 8162 locus with G/A polymorphism between SLE patients and controls (Chi2 = 8.55, chi2 = 11.85, P<0.05). The frequencies of genotype for 7872 locus with types of C/C, C/T and T/T in SLE patients were 36.1% 41.8% and 22.1%, while in control they were 51.7%, 35.0% and 13.3%, respectively. There was significant difference in the frequency of mutation in 7872 locus between SLE patients and controls (chi2 = 7.411, P<0.05). The genotype frequencies of PDCD1 8162 locus G/G, G/A and A/A in SLE patients were 50.1%, 20.3% and 28.6%, while in control they were 57.6%, 20.8% and 22.6%, respectively. There was significant difference in the frequency of mutation in 8162 locus alleles between SLE patients and controls (chi2 =7.547, P<0.05). The genetypeof PDCD1-7809(C/T) and PDCD1-8162(G/A) seemed to have the function of preventing the development of SLE ( OR = 0.583, OR = 0.485). CONCLUSION: The genotype of PDCD1 gene 7809 locus was G/G in all indigenous Han Chinese, while the SNPs of PDCD1 gene 7872 locus and 8162 locus might affect the susceptibility to SLE development. PMID- 16471223 TI - [Study on the intercellular molecule-1 polymorphisms in an Chinese population with myocardial infarction]. AB - OBJECTIVE: To investigate the association between intercellular adhesion molecule 1 (ICAM-1) gene + 12959G/A and + 13848A/G polymorphism and myocardial infarction (MI). METHODS: Polymerase chain reaction-sequence specific primers (PCR-SSP) technology and PCR-restriction fragment length polymorphism(PCR-RFLP) were used for the detection of ICAM-1 genotypes in 165 patients with MI(including acute MI and old MI) and 199 healthy controls. Plasma lipid levels and hyper sensitive C reactive protein (quantitive ELISA method was used) levels were measured in all subjects. RESULTS: + 12959G/A polymorphism was not detected in our study. The frequencies of AA,AG and GG Genotypes of + 13848A/G were 58.2% and 45.7%, 37.0% among patients and 45.2%, 4.8% and 9.0% among controls,respectively. There were statistically significant differences in the distributions of the genotype frequencies (P<0.05) between two groups, and the relative risk suffered from MI of AA genotype was 1.651 times of the GG and GA genotype (OR = 1.651, 95% CI: 1.089-2.504). CONCLUSION: There was no + 12959G/A polymorphism found in Chinese people. The coding single nucleotide polymorphism + 13848 A/G in the exon 6 of ICAM-1 gene was associated to MI and the allele A might serve as a risk factor for MI in Chinese. PMID- 16471224 TI - [Study on the molecular epidemiology of Mycobacterium tuberculosis in Shanghai]. AB - OBJECTIVE: To explore the molecular-epidemiology of Mycobacterium tuberculosis in Shanghai. METHODS: Drug-resistant and drug-susceptible strains of M. tuberculosis were randomly selected from the bank of M. tuberculosis of Shanghai Municipal Center for Disease Control and Prevention and were genotyped by mycobacterial interspersed repetitive units(MIRU) and Spoligotyping methods. The genotyping results were analyzed and combined with epidemiological data. RESULTS: The Spoligotyping results demonstrated that 89 % (81/91) of the strains belonged to the Beijing genotype. Of the patients who had received BCG-vaccination,88.5% (54/61) infected with strains of Beijing genotype and 90.0% (27/30) of the patients were not BCG-vaccinated. However, the difference was not statistically significant. Drug-resistant rate from those strains of Beijing genotype was 45.7 (37/81), lower than that of non-Beijing genotype (60.0% ,6/10). Again,the difference was not statistically significant. The MIRU results showed that 62.6 % (57/91) were strains of clusters. CONCLUSION: The Beijing genotype of M. tuberculosis were found to be the dominant strains in Shanghai. The associations between Beijing genotype strains and BCG vaccination or drug-resistant were not found. Results from cluster analysis suggested that some cases might belong to the newly developed cases. PMID- 16471225 TI - [Study on the associated risk factors of rheumatic pain: a ILAR-China, WHO COPCORD Shantou study]. AB - OBJECTIVE: To validate the difference on the prevalence of rheumatic symptoms between north and south parts of the country and to study the associated risk factors of rheumatic complaints in Shantou city, China. METHODS: A total number of 10 638 people at above 16 years of age from four samples, were surveyed in 1987, 1992, 1995, and 1999. Protocol of International League Against Rheumatism (ILAR)-China collaborative study or WHO-ILAR community oriented program for control of rheumatic disease(COPCORD) core questionnaire was implemented. Data on related rheumatic symptoms were collected and analyzed. RESULTS: The prevalence rate of rheumatic complaints had been increasing in Shantou area during the last decade (in 1987 11.6%, 1992 12.5%, 1995 16.0% and 1999 19.8%) but still lower than that seen in Beijing, China in 1987(40.0%). Rheumatic symptoms were more prevalent in females than in males, in elderly than in young people with most frequently involved site happened in low back area, followed by knee and neck. Lumbar pain was more frequent among rural residents, while neck pain was more prevalent in urban school students. Prevalence of knee pain was significantly higher in people living in multi-storey buildings without elevator than those living in single-storey houses. The peak values of Bone Mineral Density in Shantou population were (0.839 +/- 0.085) g/cm2 in men, and (0.723 +/- 0.064) g/cm2 in women, significantly higher than that reported in 13 other provinces and cities including Beijing. The sense of seeking for medical help was higher in population with higher prevalence of rheumatic symptoms than that in lower complaints group. However, no significant difference was found in the rates of disability among the different populations. CONCLUSION: The prevalence rate of rheumatic complaints was lower in Shantou than that in Beijing. Socio-economic status, environment, sex, age, occupation, ergonomics, bone mineral density, and awareness of seeking medical care might have been the associated risk factors to the prevalence of rheumatic complaints. PMID- 16471226 TI - [A seven-year follow-up analysis on prognosis factors of stroke patients]. AB - OBJECTIVE: To describe survival conditions of patients with stroke and to analyze the factors associated with survival, a seven-year follow-up study was carried out. METHODS: From Nov. 1995 to Dec. 1996, 189 stroke patients were selected in the Department of Neurology, General Hospital, Tianjin Medical University. Cases were followed up since the onset of stroke. Data collected would include case history, illness and survival conditions. Kaplan-Meier methods were used for survival description. Cox regression was used for prognostic factors analyses. RESULTS: A total number of 82 patients had been dead during the period of study and among them,58 cases died from stroke. The survival rate was 79.86% in one year, 65.46% in three years and 57.46% in seven years. Factors with statistical significance that associated with survival would include: age (RR = 1.065, P < 0.001), physical exercises before stroke (RR = 0.308, P<0.001), hypertension history (RR = 1.785, P < 0.05) and stroke history (RR =2.493, P < 0.001) while factors associated with severity of the illness were: area of cerebral lesion, conditions when discharged from the hospital, rehabilitative treatment of post discharge and recurrence. We also found that social-psychical factors as rehabilitative confidence, repression, negative event, support from relatives and friends were related to survival of stroke. CONCLUSIONS: Histories on hypertension, stroke and brain injury condition were related to the rate survival on stroke. Patients persisting physical exercises before stroke had better prognosis. The survival rate of patients with recurrence was lower than those without while social psychic factors might be related to survival. PMID- 16471227 TI - [Study on the causes and risk factors on vertigo and balance disorders in 118 elderly patients]. AB - OBJECTIVE: To understand the various factors causing vertigo and balance disorders in the elderly. METHODS: 118 elderly patients (aged equal or older than 60 years of age) with vertigo or balance disorders were retrospectively analyzed through clinical symptoms, audio-vestibular function tests, X-ray, CT scan or MRI in cervical vertebras, brain and inner ears, ultrasonography, transcranial doppler (TCD) or magnetic resonance angiography (MRA) in blood vessels on head and neck. RESULTS: Of 118 patients, 70 (23%) of them suffered perip heral vestibular disorders while 29 (58%) having cerebral vertigo or dizzness, leaving 19 cases (16%) as unclassified. CONCLUSION: For elderly patients, vertigo and balance disorders were commonly caused by many kinds of peripheral and cerebral vestibular pathological disfunctions while the functional weakness of vestibular organs and systems affected by the physiological process of ageing and different concommitant diseases as well as environmental, psychogenic factors should also be considered. PMID- 16471228 TI - [The application of counter-matching design in epidemiological research]. AB - OBJECTIVE: To explore the application of counter-matching design in epidemiological research. METHODS: Through elaboration of the study about gene environment interactions in the etiology of breast cancer, methodology regarding counter-matching design and statistic methods was introduced. RESULTS: This design improved the potential for detecting gene-environment interactions for diseases when both gene mutations and the environmental exposures of interest were rare in the general population. CONCLUSION: Counter-matching appearsed to be more appropriate than most traditional epidemiologic methods for the study of interactions involving rare factors. PMID- 16471229 TI - [Incidental cystic structures in the upper abdomen: to remove or not to remove?]. AB - An abdominal cystic structure was diagnosed incidentally in two women aged 44 and 52 years, respectively. The first patient had a mesenteric cyst that was removed because of its malignant potential. The second had a duodenal duplication cyst which was not removed due to the minor risk of complications. Two other patients, a man aged 19 years and a woman aged 40 years, were referred with mechanical complaints due to a splenic cyst and a hepatic cyst, respectively. The splenic cyst was removed surgically and the hepatic cyst was treated by sclerotherapy. Cysts are fluid-filled cavities that can develop throughout the abdomen. They are often discovered by coincidence. These four patients show that cysts in the upper abdomen may be due to various disorders and that the decision to treat is based on the risk of complications, such as bleeding, rupture or malignant potential, as well as the symptoms related to the size ofthe cyst. PMID- 16471230 TI - [Management of the obstructive sleep apnoea syndrome: more knowledge required for an optimal choice of treatment modality]. AB - In the management of the obstructive sleep apnoea syndrome (OSAS), clinicians may consider various conservative, non-invasive and surgical treatment modalities. Continuous positive airway pressure (CPAP) is regarded as the treatment of choice for, especially, moderate to severe OSAS. However, due to the obtrusive nature of this conservative treatment, especially in mild manifestations ofOSAS, the effectiveness of CPAP may be compromised due to poor therapeutic acceptance and adherence. Over the last decade, oral appliance therapy has emerged as an increasingly popular treatment alternative to CPAP. However, due to the methodological limitations of most trials studying this dental treatment modality, the precise indication for oral appliance therapy is still indistinct. In addition, based on the current level of evidence, most surgical interventions in OSAS should generally be reserved for patients with failing CPAP therapy. In order to determine whether the available treatment alternatives are competitive with CPAP, methodologically sound and preferably randomised studies are indicated. PMID- 16471231 TI - [The pathogenesis of inflammatory dermatoses, especially psoriasis]. AB - The skin contains a variety of cell types and mediators, which together constitute the skin's immune system and play a key role in protecting the human body against dangers from outside. Dysregulation of the skin's immune system, however, frequently occurs and can result in undesirable inflammatory processes in the skin. A typical example of an undesirable inflammation in the skin is the chronic inflammatory skin disease psoriasis. In the pathogenesis of psoriasis, both genetic and environmental factors play a key role. In psoriasis, the complex interactions between T-lymphocytes, antigen-presenting cells, keratinocytes and pro-inflammatory cytokines and chemokines are disturbed. The two most widely accepted hypotheses are: (a) psoriasis is a T-cell mediated autoimmune disease, and (b) psoriasis is the result of a too finely adjusted system for regulating inflammation in the skin. The result of both mechanisms is a chronic inflammatory reaction fuelled by pro-inflammatory type-I cytokines that lead to the psoriasis skin phenotype. With the development ofbiologicals, it has become feasible to target specific molecules in the immune process, for example type-I cytokines and the molecules present on pathogenic T-cells. This approach has already proved successful in the treatment of rheumatoid arthritis and Crohn's disease, creating novel therapeutic options for psoriasis and other inflammatory dermatoses. PMID- 16471232 TI - [Lung transplantation with the aid of living donors]. AB - In certain centres, following the practice in liver and kidney transplantation, lung transplantation programmes have been set up in which two healthy living donors undergo removal of the right lower lobe and the left lower lobe, respectively, which are then implanted bilaterally in the recipient in place of the usual whole right and left post-mortem lungs: 'living donor lobar lung transplantation'. The relatively high morbidity in the donors and the complicated procedure constitute obstacles to worldwide application. Besides medical problems, the procedure also raises ethical questions. Whether living donor lobar lung transplantation should also be applied in the Netherlands can only be answered following a broad public, ethical and medical discussion. In the meantime, the lung transplant centres may analyse the prerequisites and make the necessary preparations. PMID- 16471233 TI - [Recurrent miscarriage and thrombophilia]. AB - In more than half of the couples with recurrent miscarriage, no underlying cause can be found. Both acquired and hereditary risk factors for venous thrombosis, i.e. thrombophilia, constitute an aetiologic factor in a minority of patients with recurrent miscarriage. In women with a diagnosis of antiphospholipid antibody syndrome, treatment with low-dose acetylsalicylic acid and low-molecular weight heparin has been shown by a valid randomised controlled trial to improve the results of a subsequent pregnancy. However, it is unknown whether anticoagulant treatment of women with recurrent miscarriage and hereditary risk factors for thrombophilia improves the outcome of a subsequent pregnancy. PMID- 16471234 TI - [From gene to disease; primary erythermalgia--a neuropathic disease as a consequence of mutations in a sodium pump gene]. AB - Primary erythermalgia is a rare autosomal dominant inherited disorder characterized by recurrent attacks of red, warm and painful burning extremities. The gene involved in primary erythermalgia, SCN9A, encodes for a voltage dependent sodium channel alpha subunit (NaV1.7). NaV1.7 is located in dorsal root ganglions and in nociceptive peripheral neurons. It has been hypothesized that mutations lead to a gain of function and hyperexcitability of peripheral sensory neurons contributing to symptoms in primary erythermalgia. PMID- 16471235 TI - [Diagnostic image (259). A man with knee pain]. AB - In a 23-year-old man with knee pain a symptomatic bipartite patella was diagnosed; the small lateral fragment was surgically removed. PMID- 16471236 TI - [Hyoidothyroidopexy as a surgical treatment for obstructive sleep apnoea syndrome]. AB - OBJECTIVE: To evaluate the results ofhyoidothyroidopexy (HTP) in patients with obstructive sleep apnoea syndrome (OSAS). DESIGN: Prospective, descriptive. METHOD: Data were collected on patients with moderate to severe OSAS that underwent HTP between 1 March 2000 and 30 June 2004 in the St. Lucas Andreas Hospital in Amsterdam. All patients had an obstruction at the level ofthe base ofthe tongue and, to a lesser degree, the palate and either refused the use of continuous positive airway pressure (CPAP) or were unable to tolerate this treatment. The following scores were calculated preoperatively and 3 months after the operation: the apnoeahypopnoea index (AHI), apnoea index, Epworth sleepiness scale, desaturation index and visual analogue scores for hypersomnolence, snoring and pain sensation. RESULTS: During the period under investigation, 31 patients underwent HTP: 29 men and 2 women. Of these, 14 had never been operated before at the oral or hypopharyngeal level (average age: 47 years; average BMI: 26.3) and 17 had undergone unsuccessful uvulopalatopharyngoplasty (average age: 46.3 years; average BMI: 27.7). Of the 31 patients, 16 (52%) had a significant decrease in the AHI, especially those in the primary HTP-group (10/14 or 71% vs. 6/17 or 35% in the secondary group, p > 0.05). The average AHI decreased from 32.9 to 11.9 (p = 0.006) in the primary group and from 31.5 to 26.2 (p = 0.06) in the secondary group. Most of the secondary outcome measures improved statistically significantly in both groups. In one patient, tracheotomy was performed due to postoperative bleeding that threatened to obstruct the respiratory tract. CONCLUSION The short-term results of HTP as the primary treatment in patients with moderate to severe OSAS could be characterised as favourable. PMID- 16471237 TI - [The value of integrated PET-CT images in 2 lymphoma patients with skeletal localisations]. AB - In two women with Hodgkin's disease, 36 and 34 years of age, the PET-scan showed increased FDG-uptake in regions where the CT-scan did not reveal any abnormalities. Integration of the PET-CT images visualised bone marrow localisations in both patients. One patient underwent a CT-guided bone biopsy that confirmed this localisation. In both cases, the results of the integrated PET-CT images altered the therapy that would have been given on the basis ofthe standard staging technique. Both patients underwent radiotherapy. After 6 months, one patient had no visible lesions. The other patient died due to progression ofthe disease. Integrated PET-CT images can play an important role, not only in the precise classification and staging of lymphoma but also at the start of therapy, as an initial scan, in the evaluation of the response to treatment, and in the early detection of recurrence. PMID- 16471238 TI - [A boy with cholera from India]. AB - A 7-year-old Indian boy travelling from India to the United Kingdom was brought to the Emergency Clinic of Airport Medical Services at Schiphol airport in Amsterdam, the Netherlands. He had had watery diarrhoea in the aircraft and had lost consciousness. In view of the strong indications for cholera and the rice water-like diarrhoea, he was admitted to the paediatric ward of the VU Medical Centre where intravenous rehydration was carried out. He recovered within three days. A large number of comma-shaped, motile, Gram-negative rods were found in the faeces. After two days, the faeces culture revealed Vibrio cholerae O1 El Tor, serotype Inaba. On the day of the flight, the patient had drunk a litre of water from a bottle that later turned out to have been from the New Delhi water supply. Cholera is rare as an import disease in the Netherlands. Due to the severe dehydration, the infection can run a serious course and even be fatal. The infection is not transmitted from person to person. Therefore, no special measures are needed when a patient with cholera is admitted to hospital. PMID- 16471239 TI - [Thromboprophylaxis in general surgical practices in the year 2004]. PMID- 16471240 TI - [Thromboprophylaxis in general surgical practices in the year 2004]. PMID- 16471241 TI - [Abscess formation due to lost stones during laparoscopic cholecystectomy]. PMID- 16471242 TI - [A time of change--part two]. PMID- 16471243 TI - [Epidemiological importance of humans and domestic animals as reservoirs of verocytotoxin-producing Escherichia coli]. AB - BACKGROUND/AIM: A "new" pathogenic agent, verocytotoxin--producing Escherichia coli (VTEC) emerged in the last 20 years, causing an increased number of sporadic cases, as well as of outbreaks of diarrhoeal diseases. Humans and animals can be infected with VTEC, but their epidemiological importance as a reservoir of this agent is not quite clear, especially in the Balkan region. Therefore, the aim of this study was to investigate the frequency of isolation of VTEC from the intestinal tract of humans and animals and to determine the serogroups of the isolated strains. METHODS: A total of, 3 401 stool samples from humans and 2 660 samples from five different species of domestic animals were tested for the presence of this pathogen. RESULTS: VTEC was isolated from 20 (0.6%) humans stools and from 431 (16.2%) animal fecal samples (p < 0.001). Only 15 (3.3%) VTEC strains belonged to human infection-associated serogroups (O26, O55, O111, O128 and O157), designated as enterohaemorrhagic E. coli (EHEC). The most known serogroup-O157 was identified in 6 (1.3%) of the isolated VTEC strains; of them, 1 (5%) was of human origin and 5 (1.2%) were animal strains. CONCLUSION: This study revealed that domestic animals were a more important reservoir of VTEC than humans, and that the isolated VTEC strains rarely belonged to O157, as well as to other EHEC serogroups that might explain rare sporadic cases and the absence of epidemic occurrence of diarrhoeal diseases caused by VTEC in this geographic region. PMID- 16471244 TI - [Evaluation of the degree of clinical rheumatoid arthritis activity based on the concentrations of cytokines TNF-alpha, IL-12, IL-15, and IL-18 in serum and synovial fluid]. AB - BACKGROUND/AIM: Experimental in vitro and in vivo investigations in a mouse model have proved that TNF-alpha, IL12, IL-15 and IL18 participate in the pathogenesis of erosive inflammatory arthritis. The aim of this research was to determine the clinical significance of cytokines in the evaluation of the activity of rheumatoid arthritis (RA). METHODS: Inside a 4-year period we followed-up 64 patients with RA as newly ocurred or in the phase of worsening. We observed the clinical manifestation of the disease upon which we divided the patients in to 3 groups: the patients with low active RA, patients with moderate active RA, and the patients with wild active RA. The control group (n=25 patients) included the patients with osteoarthrosis (OA), and arthritis of the knee. In the samples of serum of all of the patients the concentratin of cytokines TNF-alpha, IL-12, IL 15, and IL-18 were determined using the immunoenzymatic methods in mice for human interleukines. By comparing the concentrations in 30 patients with the high, 14 patiens with moderate, and 20 patiens with the mild activity of RA it was determined that the patients with the high degree of the disease activity, had significantly high (p < 0.01; p < 0.05) concentrations of the examined cytokines in blood and synovial fluid as compared to the patients with the moderate and mild active disease. There was a relationship (p < 0.01) between the concentrations of cytokines in blood and synovial fluid with the quantity of the Disease Activity Score in 28 joints. CONCLUSIONS: Cytokines concentrations could be good indicators of the degree of the general activity of RA. This research could contribute to the interpretation of insufficiently well known views of the pathogenesis role and significance of citokines in an active disease. PMID- 16471245 TI - [Dynamic changes in calcium and phosphate plasma concentrations in the patients on peritoneal dialysis]. AB - BACKGROUND/AIM: The disturbances of active forms of vitamin D synthesis and disturbances in calcium and posphate metabolism develop early in chronic renal failure, when creatinine clearance is about 30 ml/min. Chronic hemodialysis and peritoneal dialysis only partially correct the biochemical enviroment of patients on chronic renal replacement therapy because of end-stage renal disease. These dialysis modalities can't signifficantly affect the endocrine disturbances of chronic renal failure and they have minimal modulatory effect. The management of disturbed calcium (Ca) and phosphate (P) metabolism and the maintainance of Ca x P product below 4.4 mmol/l thanks to the use of dialysate solutions with the appropriate calcium concentration and the careful dosage of phosphate binders, calcium and active vitamin D metabolits, are extremely important for the prevention of renal osteodystrophy, secondary hyperparathyroidism as well as low bone turnover disease. The aim of the study was to analyze the plasma levels of calcium, phosphate, albumin, alkaline phosphatase and parathormon (PTH) in 58 patients who were treated with continuous ambulatory peritoneal dialysis (CAPD) from March to August 2003. The use of phosphate binders and the substitution with active vitamin D metabolits were also analyzed. METHODS: We examined 58 patients, 30 males and 28 female, mean-age 52 years (range, 26-78 years), affected by end stage renal disease of the different leading cause. The average time on peritoneal dialysis program was 20 months (2-66 months). Most of the patients were treated by CAPD, while only few of them performed authomatic, cyclic or intermitent peritoneal dialysis. Most of the patients used a dialysate with 1.75 mmol/l calcium concentration. RESULTS: The study showed that our patients on chronic CAPD program during several months had normal calcemia, phosphatemia and the level of alkaline phosphatase, and that they had Ca x P product in the recommended range. PTH serum level ranged from 16 to 490 pg/l in our patients. CONCLUSION: The study showed that a balanced diet and a correct dosage of phosphate binders, as well as a careful substitution with active vitamin D metabolits render a good control of calcium and phosphate serum balance, as well as an effective prevention of renal osteodystrophy development in the patients on chronic peritoneal dialysis treatment. PMID- 16471246 TI - [Significance of the initial cytomorphological and immunocytochemical findings and the correlation with the International Prognostic Index for the survival in patients with non-Hodgkin's lymphoma]. AB - BACKGROUND/AIM: Fine-needle aspiration biopsy is a quick, economical, and safe initial method in managing a patient with suspected lymphoma. According to a few reports on this preoblem, the aim of this study was to compare histological findings to cytomorphological ones in needle aspirates. We also compared these findings to the overal survival (OS) time. METHODS: We analyzed the fine-needle aspiration biopsies of peripheral lymph nodes, and the International Prognostic Index (IPI) in 81 patients with non-Hodgkin's lymphoma (NHL). We put these findings into correlation with OS time. RESULTS: According to the International Working Formulation (IWF) criteria, the dominant cell population was asfollows: 18 patients had the small cell population, 21 patients had small cleaved cells, 18 patients had the mixed cell population, 21 patients had large cell population, 2 patients had Burkitt lymphoma type, and 1 patient had the dominant lymphoblasts. On presentation, 32 patients had a low IPI index, 32 patients had a low intermediate, and 17 patients had a high intermediate IPI. We confirmed the statistical significance (Kaplan-Mayer) of cytomorphology (p = 0.013) and IPI index (p = 0.016) for survival time. During a 48-month follow-up, OS was 37.2 months for the patients with the dominant small cells, and 32 months for the patients with small cleaved cells (PH equivalent to indolent NHL). For the patients with the dominant mixed cell population, large cell population and Burkitt limphoma cell, OS were 17, 14.4, and 9.3 months, respectively (PH equivalent to aggressive NHL). Patients with low IPI had the highest OS, 36 months for the low intermediate and only 11.6 months for the high intermediate IPI index. CONCLUSION: We concluded that an initial cytological and clinical profile of patients with NHL, might give a quick and relevant information for planning an adequate therapy. PMID- 16471247 TI - [High doses of immunoglobulin G in the therapy for severe forms of myasthenia gravis and Guillain-Barre syndrome]. AB - BACKGROUND/AIM: High doses of immunoglobulin G (IVIG) have been recognized as a very important therapeutic modality in the treatment of neurological diseases. The aim of this report was to present our experience in the treatment of severe forms of myasthenia gravis (MG) and Guillain-Barre syndrome (GBS). METHODS: We analyzed the efficacy and safety of immunoglobulin G therapy in 53 patients with severe forms of myasthenia gravis, and 27 patients with very severe forms of Guillain-Barre syndrome. RESULTS: At the end of the follow-up period, a significant improvement was noticed in 47 out of 53 patients with myasthenia gravis (88.7%). In the group of 27 patients with severe forms of Guillain-Barre syndrome an improvement was registered in 19 patients (70.3%). The side effects of this therapy were mostly mild, manifested as headache, myalgia, skin rash, adynamia, and other clinically insignificant effects. No severe side effects were recognized. CONCLUSIONS: Our study clearly demonstrated the high efficacy of IVIG therapy in the treatment of severe forms of myastehnia gravis and Guillain-Barre syndome. PMID- 16471248 TI - [Examination of cardiovascular function variables in tooth extraction under local anesthesia]. AB - BACKGROUND/AIM: Local anesthesia is the one of the most used procedures in surgical practice. It is used for toot extraction to produce analgesic and anesthetic effects. However, there is a question if it is equally safe to apply a local anesthetic combined with a vasoconstrictor (adrenaline) in healthy persons, and in the patients with a certain cardiovasccular system disease. The aim of this stady was to determine whether there were differences in cardiovascular variables during tooth extraction in healthy persons, and in cardiovascular patients when an anesthetic was applyted with adrenaline, or without it. METHODS: The examinees were divided into the group with cardiovascular diseases (CV, n=57) of H and III type, according to the American Society of Anesthesiologists (ASA) qualification, and healthy persons (H, n=55). Both groups were randomly divided into two subgroups: CVa and Ha--where the anesthetic solution had the vasoconstrictor (3% lidocaine, and 1: 100,000 adrenaline); CVb and Hb--where the anesthetic solution was without the vasoconstrictor (3% lidocaine). During the preparation for tooth extraction, the application of anesthetics, extraction and relaxation puls (fc), systolic (TAs) and diastolic arterial blood pressure (TAd) and ECG were registered. RESULTS: The values of fc did not significantly differ among the groups in any measured term. The values of systolic and diastolic blood pressure in the groups CVa and CVb were significantly higher in all the terms of measuring (p < 0.05) from the values in the groups Ha and Hb. A significant increase of TAs was registered only in the phase of tooth extraction in the CVa and CVb group (< 0.05). The values of TAd did not significantly differ between the groups in all the measured terms. Extrasystolic beats were registered in 11 patients of the CV group and in 7 patients of the H group in the phase of anesthetic application or tooth extraction. CONCLUSION: This research shoved that tooth extraction under local anesthesia did not cause complications in cardiovascular patients who were in II or III ASA qualification group, regardless the presence of a vasoconstrictor in the local anesthetic solution. PMID- 16471249 TI - [Factors which influence postoperative mortality after hip fracture]. AB - BACKGROUND/AIM: One of the most significant predictors of mortality after hip fractures is cognitive impairment (dementia). The aim of this study was to report the results of a prospective study of the influence of some factors on six-month mortality in elderly patients with hip fractures. METHOD: The elderly patients with hip fracture were assessed on admission to the hospital using the measures of cognitive function, the mobility before the fracture, and physical comorbidity, the type of fracture and the place of the injury. Six months later, we checked how many of them were still alive. RESULTS: We performed univariate and multivariate analyses in 132 patients and found that the most significant predictors of six-month mortality were dementia, comorbidity, and prefacture mobility. CONCLUSIONS: A comprehensive physical and mental health assessment of an elderly patient after hip fracture could predict mortality. A good examination of cognitive functioning could be very useful in choosing the optimal treatment for this type of patients. PMID- 16471250 TI - [Alternative and complementary medicine]. PMID- 16471251 TI - [Significance of the spatial reconstruction based on mathematical modeling in the surgical treatment of giant intracranial aneurysms]. AB - BACKGROUND: The use of computer models for the 3-dimensional reconstruction could be a reliable method to overcome technical imperfections of diagnostic procedures for the microsurgical operation of giant intracranial aneurysms. CASE REPORT: We presented a case of successfully operated 52-year-old woman with giant intracranial aneurysm, in which the computer 3-dimensional reconstruction of blood vessels and the aneurysmal neck had been decisive for making the diagnosis. The model for 3-dimensional reconstruction of blood vessels was based on the two 2-dimensional projections of the conventional angiography. Standard neuroradiologic diagnostic procedures showed a giant aneurysm on the left middle cerebral artery, but the conventional subtraction and CT angiography did not reveal enough information. By the use of a personal computer, we performed a 3 dimensional spatial reconstruction of the left carotid artery to visualize the neck of aneurysm and its supplying blood vessels. CONCLUSION: The 3-dimensional spatial reconstruction of the cerebral vessels of a giant aneurysm based on the conventional angiography could be useful for planning the surgical procedure. PMID- 16471252 TI - [Benign paroxysmal torticollis in infancy]. AB - BACKGROUND: Benign paroxysmal torticollis (BPT) is an episodic functional disorder of unknown etiology, characterized by the periods of torticollic posturing of the head, that occurs in the early months of life in healthy children. CASE REPORT: We reported two patients with BPT. In the first patient the symptoms were observed at the age of day 20, and disappeared at the age of 3 years. There were 10 episodes, of which 2 were followed by vomiting, pallor, irritability and the abnormal trunk posture. In the second patient, a 12-month old girl, BPT started from day 15. She had 4 episodes followed by vomiting in the first year. Both girls had the normal psychomotor development. All diagnostical tests were normal. CONCLUSION: The recognition of BPT, as well as its clinical course may help to avoid not only unnecessary tests and the treatment, but also the anxiety of the parents. PMID- 16471253 TI - [Acute serous meningitis in a patient with erythema infectiosum]. AB - BACKGROUND: Clinical manifestations caused by parvovirus B19 (PVB19) are various and depend on the age and immunity of an infected person. In children, the most frequent clinical manifestation of parvovirus B19 primary infection is erythema infectiosum (EI). CASE REPORT: In this case report we presented a 12-year-old patient with 2 clinical syndromes: erythema infectiosum and serous meningitis. Erythema infectiosum was manifested as fever, typical skin lesions ("slapped cheeks"), erythematous macules and papules confluent with reticular appearance on the extremities and the trunk. Serous meningitis had a mild course with an increased number of lymphocytes (120/ mm3) and the mildly increased level of proteins (0.75 g/l). The serological examination showed the presence of IgM and IgG antibodies against parvovirus B19 in serum, as well as in cerebrospinal fluid (CSF). The reduction of serum/CSF ratio of IgG antibodies was present. The symptomatic therapy was used in the treatment. The course and the prognosis were benign. CONCLUSION: Human PVB19, although non-specifically associated with CNS diseases could be an etiological factor that might cause serous meningitis. So, it should be considered in different diagnosis in patients with aseptic meningitis, especially during the outbreaks of erythema infectiosum. PMID- 16471254 TI - [Salerno]. PMID- 16471255 TI - Expression of p53 protein in gastric adenocarcinoma. PMID- 16471256 TI - [Programmed cell death: history and future of a concept]. AB - Cell death was observed and understood since the 19th century, but there was no experimental examination until the mid-20th century. Beginning in the 1960's, several laboratories demonstrated that cell death was biologically controlled (programmed) and that the morphology was common and not readily explained (apoptosis). By 1990 the genetic basis of programmed cell death had been established and the first components of the cell death machinery (caspase 3, bcl 2 and Fas) had been identified, sequenced, and recognized as highly conserved in evolution. The rapid development of the field has given us substantial understanding of how cell death is achieved. However, capitalizing on our knowledge for therapeutic purposes requires us to learn much more about how a cell commits to death, as well as recognizing that apoptosis may be the most common and efficient means of death, but that there are alternative pathways that can result in cell death even when the conventional pathway is blocked. Interestingly enough, many of the arguments and missteps in the history of the field were anticipated by Claude Bernard, and his warnings and recommendations remain valid today. PMID- 16471257 TI - [Selective "death programs" or pleiotropic"life programs"? Looking for programmed cell death in the light of evolution]. AB - "Nothing in biology makes sense except in the light of evolution", wrote Theodosius Dobzhansky, one of the founders of the Modern Synthesis that led to the unification of evolutionary theory and genetics in the midst of the 20th century. Programmed cell death is a genetically regulated process of cell suicide that is central to the development, homeostasis and integrity of multicellular organisms. Conversely, the dysregulation of mechanisms controlling cell suicide plays a role in the pathogenesis of a wide range of diseases. While great progress has been achieved in the unveiling of the molecular mechanisms of programmed cell death, a new, and somehow puzzling level of complexity has recently begun to emerge, suggesting i) that several different self destruction pathways may exist and operate in parallel in our cells, and ii) that molecular effectors of cell suicide might also perform other functions unrelated to cell death induction and crucial to cell survival, such as cell differentiation, metabolism, and the regulation of the cell cycle. These new findings, with important physiopathological and therapeutic implications, seem at odds with the paradigm of programmed cell death derived from the studies of Caenorhabditis elegans, which led to the concept of the existence of selective, bona fide death genes that emerged and became selected for their sole capacity to execute or repress cell death. In this review, I will argue that this new level of complexity might only make sense and be understood when considered in a broader evolutionary context than that of our phylogenetic divergence from C. elegans. A new view of the regulated cell death pathways emerges when one attempts to ask the question of when and how they may have become selected during a timeline of 4 billion years, at the level of ancestral single-celled organisms, including the bacteria. I will argue that there may be no such thing as a bona fide genetic cell death program. Rather, in the framework of a model that I have termed the "original sin" hypothesis, I have proposed the existence of an initial pleiotropy of the molecular tools involved in the control and execution of self-destruction- an ancestral involvement in both pro-life and pro-death activities. I will discuss how this hypothesis may be reconciled with the C. elegans paradigm of programmed cell death. Finally I will discuss how an ancestral level of pleiotropic functions of the molecular tools involved in the control of cell death, aging and genetic diversification might have favored their initial selection, their constant availability for de novo selection, and their progressive propagation in most--if not all--species during the course of evolution. PMID- 16471258 TI - [Genetic approaches to molecular mechanisms of programmed cell death in Dictyostelium]. AB - Caspase-independent cell deaths have been observed in many species including the human. However, the molecular mechanisms which govern them are largely unknown. Our present work makes use of a model organism, the protist Dictyostelium discoideum, which displays a caspase-independent cell death during its development. In rich medium, Dictyostelium multiplies vegetatively as a unicellular organism, but in starvation conditions, Dictyostelium cells aggregate, differentiate and morphogenize into a multicellular structure, called sorocarp, containing a mass of spores supported by a stalk. Cells in the stalk are considered dead on the basis of non-regrowth in a rich medium and are vacuolized. This programmed cell death is therefore developmental and vacuolar, and in addition, caspase-independent since the Dictyostelium genome does not contain caspases genes. In order to study in detail this cell death without induction of development, an in vitro experimental protocol has been adopted, which enabled us to describe the cascade of morphological events during this cell death. An insertional mutagenesis approach, followed by appropriate selection or screening of mutants potentially resistant to death, attempted at establishing the cascade of molecular events leading to vacuolar death of Dictyostelium cells. A better understanding of alternative death pathways may allow to control different types of cell deaths in the cases of cancers or neurodegenerative diseases. In this short review, we will discuss briefly some generalities about the development of Dictyostelium in starvation conditions, and we will focus on the course of programmed cell death in Dictyostelium and on the genetic tools used to elucidate the corresponding molecular mechanisms. PMID- 16471259 TI - Immunopathological consequences of the loss of engulfment genes: the case of ABCA1. AB - Programmed cell death plays a crucial role in the maintenance of cell homeostasis. An initial, effector phase leads to the generation of apoptotic corpses and is closely followed by a swift clearance by professional or amateur phagocytes. Several aspects distinguish this latter process of engulfment of dying cells from the classical forms of phagocytosis. They concern all aspects of the process from the recognition of the prey to the final outcome, i.e. immunological silence. The engulfment of dead cells is a process highly conserved through evolution and it has been studied in parallel in two systems, mammalian cells and the nematode C. elegans. ABCA1 and its ortholog CED-7 in the nematode are key players of engulfment. Their mode of action is somehow original in the panorama of engulfment receptors since they act as lipid transporters. While in the worm the loss of CED-7 has phenotypic consequences exclusively on engulfment, in the mouse the deletion of ABCA1 by homologous recombination has highlighted broad consequences on macrophage biology. Among those we will discuss here the aberrant responses of ABCA1-/- mice to Plasmodium berghei ANKA infection, concerning in particular the development of cerebral malaria (CM), a cytokine induced immunopathology. This syndrome involves a central role of monocytes and, as shown recently, high levels of circulating microparticles. It was found that ABCA1 loss completely protects against CM and its associated mortality. This observation, together with the demonstration of quantitative and functional modifications of microparticles, suggests that microparticles may be involved in CM pathogenesis. The ABCA1 transporter thus appears to control susceptibility to CM, thereby providing new insights in its pathophysiological mechanisms and potential new therapeutic avenues. PMID- 16471260 TI - [Changes in the outer mitochondrial membranes during apoptosis]. AB - Mitochondria are involved in many apoptotic responses. Following permeabilization of their outer membrane, they release many apoptogenic proteins, including cytochrome c, which contribute to caspase activation. The mechanisms responsible for membrane permeability are not completely understood. Here, we briefly review the mechanisms that have been proposed to explain this phenomenon. PMID- 16471261 TI - [Dependence receptors DCC and UNC5H: the role of apoptosis in the control of tumorigenesis]. AB - Recent studies have led a different view about membrane receptors. While a receptor used to be considered as inactive until bound by its ligand, it has been proposed that some receptors may also be active in the absence of their ligand. These so-called dependence receptors induce a specific death signal when the ligand is absent from the cell. Therefore, the expression of one of these receptors drives the cell to become dependent on the presence of the ligand for its survival. We have hypothesized that this mechanism allows inhibition of tumor growth, by inducing apoptosis of "abnormal" cells that would usually grow when ligand are unavailable--i.e., during local growth of tumor cells or growth beyond primary tumor site -. Along this line, back in the early 90s, Vogelstein and colleagues suggested that a gene called DCC (for "deleted in colorectal cancer") could be a tumor suppressor gene because it was found to be deleted in more than 70 % of colorectal cancers, as well as in many other cancers. During the last fifteen years, controversial data have failed to firmly establish whether DCC is indeed a tumor suppressor gene. However, our observation that DCC behaves as a dependence receptor that induces cell death unless its ligand netrin-1 is present, together with the fact that mice engineered to block DCC-induced cell death by overexpressing netrin-1 are predisposed to develop colorectal tumors, strengthen the role of dependence receptors as tumor suppressors. In this review, we will describe the implication of the netrin-1/receptor pairs as novel negative regulators of tumor development. PMID- 16471262 TI - [Erythropoiesis: a paradigm for the role of caspases in cell death and differentiation]. AB - Erythroid differentiation involves the transcription factor GATA-1 that positively regulates promoters of erythroid genes (including haemoglobin, glycophorin, erythropoietin receptor) and of erythropoietin. Terminal erythroid differentiation is characterized by major morphological changes that include chromatin condensation and cell size reduction. The morphological changes are partially similar at least to those observed during apoptosis. The production of red cells depends on the apoptosis rate of erythroid progenitors and precursors. Upon erythropoietin starvation or engagement of the death receptor Fas, caspases are activated in erythroid precursors and cleave GATA-1, thus inducing maturation arrest and apoptosis of immature erythroblasts. We have recently demonstrated that, upon erythropoietin stimulation, caspase-3 was also activated, an event required for human terminal erythroblast maturation. Proteins cleaved by caspases in erythroid cells undergoing terminal differentiation include Lamin B and Acinus, which are involved in chromatin condensation. In contrast, despite caspase-3 activation neither GATA-1 degradation nor apoptosis was observed. Thus, the fate of erythroid precursors is determined downstream of caspase activation by the pattern of cleaved targets. Therefore, there are some mechanisms underlying the selective protection of caspase-3 targets during erythropoiesis. This model in which caspases activation is required for differentiation may apply to other haematopoietic or non haematopoietic cellular systems which are described in this review. PMID- 16471263 TI - [Autophagy in cell survival and death]. AB - Macroautophagy hereafter referred to as autophagy is a major lysosomal catabolic pathway for macromolecules and organelles conserved in eukaryotic cells. The discovery of the molecular basis of autophagy has uncovered its importance during development, life extension and in pathologies such as cancer, certain forms of myopathies and neurodegenerative diseases. Autophagy is a cell survival mechanism during starvation that is controlled by amino acids. Starvation-induced autophagy is an anti-apoptotic mechanism. However autophagy is also an alternative to apoptosis through autophagic cell death. In many situations apoptosis and autophagy can both contribute to cell dismantlement. PMID- 16471265 TI - [Huntington's disease: intracellular signaling pathways and neuronal death]. AB - Huntington's disease (HD) is a mid-life onset neurodegenerative disorder characterized by unvoluntary movements (chorea), personality changes and dementia that progress to death within 10-20 years of onset. There are currently no treatment to delay or prevent appearance of the symptoms in the patients. The defective gene in HD contains a trinucleotide CAG repeat expansion within its coding region that is expressed as a polyglutamine (polyQ) repeat in the protein huntingtin. The exact molecular mechanims by which mutant huntingtin induces cell death as well as the function of huntingtin are not totally understood. Studying mechanisms by which polyQ-huntingtin induces neurodegeneration has shown that phosphorylation plays a key role in HD. The IGF-1/Akt/SGK pathway reduces polyQ huntingtin induced toxicity. This anti-apopototic effect is mediated via the phosphorylation of serine 421 of huntingtin. Moreover, components of this pathway are altered in disease. What is the function of huntingtin? Several studies indicate that huntingtin is an anti-apoptotic protein that could regulate intracellular dynamic. We recently demonstrated, that huntingtin specifically enhances vesicular transport of brain-derived neurotrophic factor (BDNF) along microtubules. Huntingtin-mediated transport involves Huntingtin-Associated Protein-1 (HAP1) and the p150(Glued) subunit of dynactin, an essential component of molecular motors. BDNF transport is attenuated both in the disease context and by reducing the levels of wild-type huntingtin. The alteration of the huntingtin/HAP1/ p150(Glued) complex correlates with reduced association of motor proteins with microtubules. Finally, polyQ-huntingtin-induced transport deficit results in the loss of neurotrophic support and neuronal toxicity. PMID- 16471264 TI - [Experimental modulation of apoptosis: physiopathological and therapeutic targets]. AB - In the liver, the importance of apoptosis is not only evident during development and homeostasis of the biliary tree but plays also a prominent role in liver pathogenesis. Ligand binding to cell surface death receptors such as Fas activates the extrinsic pathway. This pathway predominates in autoimmune liver diseases, viral hepatitis, liver allograft rejection. Hepatocyte apoptosis is also significantly increased in patients with alcoholic hepatitis and nonalcoholic steatohepatitis and correlates with disease severity and hepatic fibrosis. We have used this specific susceptibility of the liver to apoptosis to develop two different approaches: 1) a cell therapy strategy based on a survival advantage to an apoptotic stimulus conferred to transplanted hepatocytes and 2) a new model of hepatocyte conditional ablation based on a controlled activation of the cell death program. PMID- 16471266 TI - [The Bcl-2 family of proteins as drug targets]. AB - Programmed cell death or apoptosis is a crucial process for normal embryonic development and homeostasis. Apoptosis is known to be coupled to multiple signalling pathways. Identification of critical points in the regulation of apoptosis is of major interest both for the understanding of control of cell fate and for the discovery of new pharmacological targets, particularly in oncology. Indeed, defects in the execution of apoptosis are known to participate in tumour initiation and progression as well as in chemoresistance. The Bcl-2 family members constitute essential intracellular players in the apoptotic machinery. Those proteins are either pro or anti-apoptotic, they interact with each other to regulate apoptosis. Inhibiting the heterodimerisation between pro- and anti apoptotic members is sufficient to promote apoptosis in mammalian cells. Small molecules, antagonists or peptidomimetics inhibiting this heterodimerisation, represent a therapeutic prototype targeting the apoptotic cascade. They induce cell death by activating directly the mitochondrial apoptotic pathway. Considerable evidence indicate that such Bcl-2 antagonists could be useful drugs to induce apoptosis preferentially in neoplastic cells. PMID- 16471267 TI - [This death which is necessary for life]. PMID- 16471268 TI - [Inhibition of phosphatidylethanolamine and phosphatidylinositol synthesis, alteration of the G0 and G1 cell cycle phases and spontaneous apoptosis in lymphocytes from patients under immunosuppressive treatment]. AB - Lymphocytes from patients treated with immunosuppressive agents were cultured for 48 hours either with or without concanavalin A. Phospholipid synthesis was then studied using 32p pulse-incorporation (5-hours pulses). Phosphatidylethanolamine synthesis was strongly decreased under immunosuppressive treatment: 1.5-fold in resting lymphocytes and 3- to 4-fold in concanavalin A-stimulated lymphocytes. Phosphatidylinositol synthesis also decreased about 2-fold in stimulated lymphocytes. These results indicate a loss of sensitivity of immunodeficient lymphocytes to the mitogen and an alteration of the G0 and the late G1 cell cycle phases. In parallel, but after a 72-hours incubation, lymphocytes were analysed by flow-cytofluorimetry with propidium iodide. Under concanavalin A-triggered stimulation, the entry into the S phase was much lower in immunodeficient lymphocytes as compared to standard. The characteristics of the G0-G1 population of lymphocytes were also modified. More importantly, after incubation in the culture medium in the absence of mitogen, we observed, among the immunodeficient lymphocytes, a high level of apoptotic cells, about 20 to 30%. This susceptibility to spontaneous apoptosis seems inherent to the status of immunodeficiency itself, whatever its origin. It may be related to the inhibition of phosphatidylethanolamine synthesis in the G0 phase. PMID- 16471270 TI - Yet another World AIDS Day. PMID- 16471269 TI - Ambulatory care visits to physician offices, hospital outpatient departments, and emergency departments: United States, 2001-02. AB - OBJECTIVE: This report presents statistics on ambulatory care visits to physician offices, hospital outpatient departments, and hospital emergency departments. Ambulatory medical care utilization is described in terms of patient, practice, facility, and visit characteristics. Office-based care is further subdivided into the categories of primary care, surgical specialties, and medical specialties. METHODS: Data from the 2001 and 2002 National Ambulatory Medical Care Surveys (NAMCS) and National Hospital Ambulatory Medical Care Surveys (NHAMCS) were combined to produce averaged annual estimates of ambulatory medical care utilization. RESULTS: Patients in the United States made an estimated 1.1 billion visits per year in 2001 and 2002 (annual average) to physician offices, hospital outpatient departments, and emergency departments, a rate of 3.8 visits per person annually. This marks the first time that the annual estimate of visits has surpassed the billion mark and is also a significant increase from the 1999-2000 estimate. The change was primarily driven by a jump in the number of visits to primary care physicians. The distribution of visits by patient age, sex, race, expected source of payment, geographic region, and whether the visit occurred in a metropolitan statistical area (MSA) varied across ambulatory care settings. Females had higher visit rates than males to all settings except office-based surgical specialists and emergency departments (ED). Black persons had higher visit rates than white persons to hospital outpatient and emergency departments, but lower visit rates to office-based surgical and medical specialists. Visits to emergency departments were more likely to be patient-paid or no charge, possibly reflecting a lack of private health insurance, than were visits to physician offices. Visit rates to office-based medical specialists were more than double in MSAs compared with non-MSAs. PMID- 16471272 TI - Cardiovascular effects of cocaine--a patient with HIV infection and a bad habit. PMID- 16471271 TI - Medicare Part D drug benefit and HIV/AIDS: questions and answers. AB - This newly enacted legislation has the potential to expand access to needed medications for persons with HIV/AIDS and actually improve care by making medication more affordable. This is particularly true for patients who are currently enrolled in ADAPs with limited or restricted formularies. The benefit of the Part D program in states with more generous ADAP formularies, such as New York and California, is not as clear. Choosing a drug plan can also be complex. Providers and case managers should have knowledge of the plans that are offered in their areas. While a program such as this has the potential to improve access to drugs, providers should expect the unexpected, especially with a newly implemented program that is wide in scope and subject to change over time. In the end, we need to work to avoid interruptions in treatment for our patients. PMID- 16471273 TI - Preventing pneumococcal disease. PMID- 16471274 TI - Antiretroviral update: further validation of the guidelines. PMID- 16471275 TI - Primary anetoderma and HIV infection: a case report. AB - Anetoderma is characterized by circumscribed areas of flaccid skin caused by the loss of elastic tissue in the dermis. It may be primary or secondary to various dermatoses. The primary form has been reported in patients with autoimmune diseases, increased levels of antiphospholipid antibodies, prothrombotic abnormalities, and recently, HIV-1 disease. The origin of anetoderma remains unknown. A case of primary anetoderma is reported in a 45-year-old man with asymptomatic HIV-1 infection who was receiving antiretroviral therapy. Laboratory research included the classic immunologic investigations and screening for prothrombotic abnormalities. Possible pathogenic mechanisms of anetoderma, especially with respect to HIV-1 infection and antiretroviral therapy, are discussed. PMID- 16471276 TI - Noninfectious cutaneous manifestations of HIV infection in children. AB - Skin infections and proliferative conditions affecting the skin commonly occur in children with HIV/AIDS and are differentiated from infections in immunocompetent children by unusual severity and distribution and age at onset. Skin manifestations of HIV disease usually are the result of various causes,from concomitant infection to nutritional deficiencies. Photographs to aid in the identification and appreciation of the atypical presentation of skin manifestations resulting from noninfectious complications of pediatric HIV infection are presented. PMID- 16471277 TI - [Tutorial for new psychiatric nurses]. PMID- 16471279 TI - [Evaluation of nursing practices in psychiatry]. PMID- 16471281 TI - [Risk-taking in psychiatry. The risk of displeasing you]. PMID- 16471282 TI - [The principle of precaution]. PMID- 16471283 TI - [Risking a deinstitutionalization project]. PMID- 16471284 TI - [Risking violence in daily life]. PMID- 16471285 TI - [Thinking about risk ... risking to think?]. PMID- 16471286 TI - [The risk of forgetting]. PMID- 16471287 TI - [Risk management in psychiatry]. PMID- 16471288 TI - [About the disaster in Sumatra]. PMID- 16471289 TI - [Guideline for nursing care in mental health. (2/2). Evolution of the nursing role]. PMID- 16471290 TI - [Value of motivational interviews in the management of alcoholic patients]. PMID- 16471291 TI - The international Surviving Sepsis Campaign--a collaborative launch in England. PMID- 16471292 TI - Scientific revolutions and the structure of critical care. PMID- 16471293 TI - Severe sepsis: patient management focusing on administration of drotrecogin alpha (activated) infusion. AB - This article presents a case study of a 49-year-old male admitted to the Intensive Care Unit (ICU) via Accident and Emergency following a collapse at home with a diagnosis of severe sepsis. The pathophysiology of sepsis is discussed together with a physiological assessment of the patient's cardiovascular, respiratory and renal systems, coagulation and metabolic responses. The discussion focuses on the period from ICU admission to commencement of drotrecogin alpha (activated) treatment (also known as recombinant human activated protein C). The management of sepsis and the treatment the patient received are outlined with particular focus on the strategic and procedural issues of the drug administration. The article concludes by presenting nursing clinical guidelines which will provide helpful guidance for nurses when caring for patients receiving drotrecogin alpha (activated) treatment. This is especially welcome, as drotrecogin alpha (activated) is internationally recommended in the treatment of severe sepsis. PMID- 16471294 TI - Body positioning and its effect on oxygenation--a literature review. AB - Evidence-based health care has become a priority in the National Health Service (NHS), with increased emphasis on clinical practices that are grounded in quality evidence rather than those that persist because of tradition. Turning and positioning of patients are well-accepted nursing activities. Appropriate positioning of the critically ill patient can dramatically improve gas exchange, resulting in a shorter stay in the critical care unit and an improved outcome. This study reviews the current published literature on the subject of positioning and gas exchange, with emphasis on oxygenation. Conclusions made from this study are that nurses need to be aware of how different positions can affect patients' oxygenation. Further research on exploring patients' experience of positioning within the critical care environment and nurses' understanding of positioning and oxygenation is needed. PMID- 16471295 TI - Implementing a protocol for weaning patients off mechanical ventilation. AB - Weaning patients from ventilation can be a costly and time-consuming intervention. This article describes how a protocol was designed and introduced into the critical care unit of a district general hospital in 2003. A step-by step approach was used based on that outlined by the Modernisation Agency and The National Institute for Clinical Excellence. The purpose of the project was to improve the weaning process in the unit by devising a protocol, which would give structure to weaning and help maintain continuity. It was hoped that the changes in practice would also reduce ventilator time and improve patient outcomes. A multi-professional group interested in weaning worked together to formulate a protocol, which was duly implemented into the unit. After implementation, the protocol was audited and subsequently adopted by the unit. Although it was agreed that the structure and the continuity of weaning had improved, reducing weaning times and patient outcomes was difficult to measure. This article explains how the protocol came to be written and how it was implemented into the unit. PMID- 16471296 TI - Intensive care nurses' beliefs and attitudes towards the effect of open visiting on patients, family and nurses. AB - The policy of family visits to patients admitted into intensive care units (ICUs) has been liberalised in recent years. This change has been progressive in our unit from a restricted to an open visiting policy. We have carried out this study based on Ajzen and Fishbein reasoned action theory. The aim of this study is to analyse the relationship between nurses' beliefs and attitudes towards the effect of an open visiting policy on patients, family and nurses. A descriptive correlational design was used. The sample included 46 intensive care nurses from a teaching hospital in Spain, who completed two self-administered anonymous questionnaires. One of them, derived from empirical results of Kirchhoff et al. (1993) and Simpson et al. (1996) studies, was a Likert-type scale including 26 items divided in three subscales (patient, family and nurses). This scale was used to obtain the nurses' beliefs regarding the effects of open visiting. The second questionnaire was a differential semantic scale (used by the previous authors), which analysed nurses' attitudes towards visiting on the patient, family and nurses. Nurses' beliefs about the effect of visiting were positive, achieving a mean value of 3.001 (patient 3.04, family 3.23 and nurses 2.78) on a scale with a maximum value of 4. The mean score obtained on the scale of attitudes toward an open visiting policy was 6.005 (patient 6.41, family 6.37 and nurses 5.22), with a maximum of 7. The correlation between beliefs and attitudes was significant and positive (r = 0523, p < 0.0001). Comparison of sociodemographic variables with beliefs and attitudes disclosed no statistically significant differences, except for the two following variables: attitude and having children (t = -2.254, p = 0.03) which obtained a higher score. There is a correlation between nurses' beliefs and attitudes regarding the positive effects of open visiting on patients, family and nurses. PMID- 16471297 TI - An evaluation of the contribution of critical care outreach to the clinical management of the critically ill ward patient in two acute NHS trusts. AB - This paper reports on an evaluation of the role and contribution of outreach in the management of the critically ill ward patient using Stake's Responsive Model (Stake, 1975) and case study methodology (Simons, 1980). Twenty cases were examined, purposefully sampling all staff involved in the case identified by an initial interview with the outreach nurse. In total, 80 interviews were carried out, 20 with the outreach nurses and 54 with other members of health care teams involved in the cases, and six further targeted in-depth interviews with senior anaesthetic and nursing staff. The outreach contribution which emerged from the data analysis consisted of four core categories: action (getting things done, getting decisions made and following through), focus and vision (concentrating on one patient and having a vision of what action was needed to meet their care needs), orchestration (a communication and co-ordinating role) and expertise (bringing critical care skills and experience to the bedside). These categories were validated and developed in the six in-depth interviews. Three themes emerged from the data describing aspects of the acute care context in which outreach operates. The interviews revealed a battleweary workforce overwhelmed by the complex and increased demands of the critically ill ward patient. The medical and nursing teams at the bedside are inexperienced and often unsupported by senior clinical decision-makers. This is dealt with by 'passing the buck' creating gaps and delays in care management which are the problems addressed by the outreach contribution. Outreach may solve problems for the critically ill ward patient, but the underlying causes remain poorly understood. PMID- 16471298 TI - Pre-registration education: making a difference to critical care? AB - This paper aims to discuss the development of a pre-registration high-dependency nursing programme and evaluate its effects on student's perceived learning and confidence in managing critically ill patients. The programme consists of two modules that compliment one another, 'Care of the Acutely III Adult' focuses on a variety of disease processes and subsequent nursing care. Whilst the second module titled 'Caring for the Highly Dependant/Critically III Adult' focuses on assessment skills related to critically ill patients. The paper explores the content and delivery of the modules including the advantages and disadvantages of implementing them. Student evaluation from 59 nursing students found that student's knowledge, assessment skills and management of the critically ill patient had improved since completing the modules. Nurse Consultants, intensive therapy unit matrons and Critical Care Outreach nurses have acknowledged an increase in the number of nursing students identifying and referring critically ill patients to outreach teams. They also stated improvements in nurse recruitment in critical care since commencement of the modules. PMID- 16471299 TI - Starting a med spa. PMID- 16471300 TI - Patient information. Folic acid: for pregnancy and more. PMID- 16471301 TI - A reimbursement action plan. PMID- 16471302 TI - Reduction of a joint dislocation. PMID- 16471303 TI - Bioidentical hormones. Achieving the perfect fit. PMID- 16471304 TI - Erasing the years. An overview of dermal fillers. PMID- 16471306 TI - From the inside out. Weight loss before or after cosmetic surgery. PMID- 16471305 TI - Aging and antioxidants. Fruits and vegetables are powerful armor. PMID- 16471307 TI - Cytomegalovirus infection. Evaluation and management in neonates. PMID- 16471308 TI - End-stage liver disease. Determining when transplant referral is necessary. PMID- 16471310 TI - Someone to lean on. Essentials of your first NP job. PMID- 16471309 TI - Something old, something new, something borrowed. COPD treatment today. PMID- 16471311 TI - NPs to the rescue. PMID- 16471312 TI - Genetic models of human cancer as a multistep process. Paradigm models of colorectal cancer, breast cancer, and chronic myelogenous and acute lymphoblastic leukaemia. AB - Tumour formations arise as a consequence of alterations in the control of cell proliferation as well as with disorders in interactions between cells and their environment that result in invasion and metastasis. Recent advances in understanding the genetic basis of malignant diseases have been dominated by research in colorectal cancer. Genetic alterations of several proto-oncogenes and tumor-suppressor genes (e.g. APC/MCC, RAS, DCC, p53 mutations and/or allelic losses, hyperexpression of c-MYC and RB genes), as well as other genomic alterations, appear at characteristic stages of tumor development and are observed in most neoplasms. Generally, the normal cell has multiple independent mechanisms that regulate its growth and differentiation potential, and several separate events would, therefore, be needed to override these control mechanisms, as well as induce the other aspects of the transformed phenotype, like metastasis. These signals may be either positive or negative, and the acquisition of tumorigenicity results from genetic changes that affect these control points following a multistep mode. Statistics of the frequency of cancer incidence with age in humans indicate that for the genesis of e.g. lung carcinoma, five or six steps are required. Other types of cancers, such as leukemias and sarcomas, probably require quite a different number of rate-limiting changes. One of the best characterized tumours to provide a genetic model is colorectal tumorigenesis. Mutations implicated in breast cancer tumorigenicity are also studied and used as a genetic model in the literature worldwide. Finally, activation of c-abl in chronic myelogenous leukaemia (CML) and acute lymphoblastic leukaemia could also be presented as an example, which provides probably the strongest evidence for the role of proto-oncogenes in human malignancy process. PMID- 16471313 TI - Efficacy of Helicobacter pylori eradication on the chronic mucosal inflammation of the remnant stomach after distal gastrectomy for early gastric cancer. AB - Although eradication of Helicobacter pylori (Hp) after early gastric carcinoma has been recommended, very limited studies have been reported and the method differs from standard therapy. Here, we attempted the eradication of Hp in the remnant stomach after surgery for primary gastric cancer with the standardized method. We examined efficacy and the safeness of the treatment. Thirty-three H. pylori-positive patients after distal gastrectomy were treated with proton pump inhibitor (PPI)-based triple therapies. After eradication, endoscopic and histological changes were classified on the basis of the Updated Sydney System. The eradication rate in the remnant stomach was 90.9% (30 out of 33 cases) after triple therapy. Temporal minor side effects were notified in 3 cases. After eradication, the remnant stomach showed significant decreases in inflammation- and activity-scores. Moreover, significant improvement in glandular atrophy to normal mucosa was found. In conclusion, PPI-based standard therapy is just as effective for Hp eradication in the remnant stomach than it is in the non operative stomach. Eradication therapy could be performed safely and resulted in a significant improvement in inflammation and atrophy of the mucosal layer in the remnant stomach after early gastric cancer surgery. PMID- 16471314 TI - MR spectroscopy of prostate cancer. Initial clinical experience. AB - Aim of the study was to evaluate the effectiveness of proton MR Spectroscopic (MRS) imaging in the detection and localization of prostate cancer, prospectively compared with histopathologic findings. Magnetic Resonance (MR) and MRS imaging were performed in 65 patients with high levels of prostate-specific antigen (PSA) and suspicious areas at the transrectal ultrasound (TRUS). At MR areas of interest were reported as normal, equivocal or suspicious. At MRS imaging, cancer was diagnosed as "possible" if the ratio of choline plus creatine to citrate exceeded 2 SDs above mean normal peripheral zone values or as "definite" if the ratio exceeded 3 SDs. All patients underwent a TRUS 10-core biopsy within 30 days of the imaging study. MR alone showed sensitivity, specificity, positive predictive values, negative predictive values and accuracy for detection of prostate cancer of 85%, 76%, 53%, 91% and 65%, respectively, whereas MRS alone showed 89%, 77%, 78%, 69% and 83%, respectively. These values were 71%, 90%, 89%, 74% and 80% when the prostate was evaluated combining MR and MRS. The addition of MRS to the MR imaging provides a higher specificity in tumour detection and can be recommended as a problem-solving modality for patients with elevated PSA levels and suspicious TRUS before biopsy. PMID- 16471315 TI - Influence of micrometastasis on N stage in gastric cancer and clinical application. AB - In order to evaluate the real extent of lymph node metastasis (LNM) in gastric cancer, an immunohistochemical (IHC) analysis was performed. We examined 11173 lymph nodes removed from 355 patients with all stages of gastric carcinoma. Tissue preparations were stained with cytokeratin 18, monoclonal antibody against cytokeratin. Micrometastases were found in 2.5% of the lymph nodes and in 31.3% of patients. The incidence of the patients with LNM increased to 9.1% in T(1m) (n = 99), 31.6% in T(1sm) (n = 95, 23.1% in sm1, 34.8% in sm2), 66.7% in T2 (n = 108, 48.8% in mp, 76.5% in ss), 88.1% in T3 (n = 42), and 90.9% in T4 (n = 11) lesions. Upstage was identified in 8.5% of patients: 6.7% in T1 (4.0% in m, 7.7% in sm1, 10.1% in sm2), 14.8% in T2 (20% in mp, 11.8% in ss), 2.4% in T3, and 0% in T4. Factors related to LNM were: tumor size and lymphatic invasion in mucosal lesions; only lymphatic invasion in submucosal lesions; size and depth of tumor, and lymphatic invasion in T2 lesions. In conclusion, the incidence of micrometastasis in regional lymph nodes was higher than we imagined in T1 lesions, more than D1 lymphadenectomy for sm1 and selected cases of mucosal cancer, and D2 lymphadenectomy for sm2 are necessary. PMID- 16471317 TI - Environmental factors related to Epstein-Barr virus-associated gastric cancer in Japan. AB - Epstein-Barr virus (EBV)-encoded small RNA can be detected in about 1-17% of gastric carcinomas. To elucidate the lifestyles and other factors related to the EBV-associated gastric carcinoma (EBV-GC), we interviewed 43 EBV-GC cases and 162 non EBV-GC cases in Kagoshima Prefecture, Japan from 1996-2001. We mainly focused on lifestyles predominant among men because of its male predominance. Although the prevalence of smokers in EBV-GC cases was higher than among non EBV-GC cases, the difference was not significant (P = 0.131). Frequent drinking of coffee and high-temperature drinks, as well as frequent intake of salty and spicy foods, were more prevalent among EBV-GC cases, but only frequent intake of salty food showed a significant difference between EBV-GC and non EBV-GC cases (P = 0.026). In addition, EBV-GC cases tended to be exposed to wood dust and/or iron filings (P = 0.068) and tar (P = 0.097). These findings, together with a high frequency of EBV-GC among remnant cancers after partial gastrectomy, suggest an association between mechanical injuries to the stomach membrane and the high frequency of EBV GC. The present study also showed that EBV-GC cases tended to be elder brothers/sisters (P for trend = 0.029) suggesting that age at primary infection with EBV may be older in EBV-GC cases than non EBV-GC cases. PMID- 16471316 TI - Feasibility of preoperative chemoradiation in rectal cancer patients aged 70 and older. AB - In the present study, we report results of 28 rectal cancer patients, aged 70 years and older, treated with preoperative radiotherapy and 5FU concomitant chemotherapy. Twenty-eight out of 136 patients treated in our Department between 1997 and 2004 aged > or = 70 years, mean 73 (range 70-81); 3 T2, 18 T3, 7 T4; 15 N0, 5N1, 8 N2; Radiotherapy (5040 cGy, 28 fractions) was delivered combined with 5FU - based concomitant chemotherapy. Compliance to chemoradiotherapy was excellent. Major acute toxicity (> or = G3) evaluation showed haematological Grade 3 only in 2 patients. No severe acute Gastrointestinal toxicity was observed. All patients underwent surgery without severe perioperative complications. Complete pathological response pT0 was found in 3 patients (11%). Overall T downstaging occurred in 61% of the cases. Mean follow up was 34 months (range 4- 84). Kaplan Meier Overall Survival and Disease Free Survival at 5 years were 74% (95% CI 54 -95) and 65% (95% CI 38-93), respectively. Only 1 patient showed G3 diarrhea according to CTCAE that interfered with his Quality of Life and required hospitalization. In conclusion, concomitant radiochemotherapy 5FU based is safe in rectal cancer patients aged > or = 70 with a good tumour downstaging (61% of patients) and excellent feasibility. No treatment related death was observed. PMID- 16471318 TI - Chemopreventive potential of Triphala (a composite Indian drug) on benzo(a)pyrene induced forestomach tumorigenesis in murine tumor model system. AB - The present work is probably the first report on cancer chemopreventive potential of Triphala, a combination of fruit powder of three different plants namely Terminalia chebula, Terminalia belerica and Emblica officinalis. Triphala is a popular formulation of the Ayurvedic system of medicine. Our findings have shown that Triphala in diet has significantly reduced the benzo(a)pyrene [B(a)P] induced forestomach papillomagenesis in mice. In the short term treatment groups, the tumor incidences were lowered to 77.77% by both doses of Triphala mixed diet. In the case of long-term treatment the tumor incidences were reduced to 66.66% and 62.50% respectively by 2.5% and 5% triphala containing diet. Tumor burden was 7.27 +/- 1.16 in the B(a)P treated control group, whereas it reduced to 3.00 +/- 0.82 (p < 0.005) by 2.5% dose and 2.33 +/- 1.03 (p < 0.001) by 5% dose of Triphala. In long-term studies the tumor burden was reduced to 2.17 +/- 0.75 (p < 0.001) and 2.00 +/- 0.71 (p < 0.001) by 2.5% and 5% diet of Triphala, respectively. It was important to observe that Triphala was more effective in reducing tumor incidences compared to its individual constituents. Triphala also significantly increased the antioxidant status of animals which might have contributed to the chemoprevention. It was inferred that the concomitant use of multiple agents seemed to have a high degree of chemoprevention potential. PMID- 16471319 TI - Involvement of hypoxia-inducible factor-1-alpha in multidrug resistance induced by hypoxia in HepG2 cells. AB - Aim of the study was to explore the influence of hypoxia on multidrug resistance related genes and the potential role of hypoxia-inducible factor-1-alpha (HIF 1alpha) in formation of multidrug resistance in HepG2 human hepatocellular carcinoma cell line. HepG2 cells were subjected to hypoxia in a cohort of exposed time. A cell model stably expressing HIF-1alpha was established by liposome mediated transfection of plasmid pcDNA3/HIF-1alpha into HepG2 cells. Apoptosis of HepG2 cells exposed to hypoxia or transfected by plasmid pcDNA3/HIF-1alpha was detected by Flow Cytometry after administration of chemotherapeutic drug (5-Fu). Real-time fluorescent quantitative PCR and Western-blot technique were used to analyze the expressions of multidrug resistance related genes mdr1, MRP1 and LRP at mRNA and protein level, respectively. Apoptosis Index of HepG2 cells exposed to hypoxia stepped down as exposed time extended after administration of 5-Fu. The expression of mdr1, MRP1 and LRP gene and protein revealed a hypoxic time dependent induction and was synchronous with the alterations of HIF-1alpha in HepG2 cells exposed to hypoxia. The expressions of these multidrug resistance related genes were remarkably increased in HIF-1alpha transfected HepG2 cells as compared to empty vector transfected cells. Apoptosis index of HIF-1alpha transfected cells was obviously less than that of control cells when they were simultaneously exposed to 5-Fu for 24hrs. In conclusion, ambient hypoxia might be one of the causes for the formation of multidrug resistance in HepG2 human hepatocellular carcinoma cell line. Hypoxia-elicited multidrug resistance related protein expression might be a pathway for resistance of HepG2 cells to chemotherapeutics and HIF-1alpha might be involved in this process. PMID- 16471320 TI - Interstitial transfer factor as adjuvant immunotherapy for experimental glioma. AB - Glioblastoma multiform (GBM) is the most common tumour of the central nervous system in humans. Unfortunately its prognosis is poor and because of the lack of efficacious therapies, immunotherapy is a potential treatment. Transfer factors (TF) are low molecular weight dialysable products extracted from immune cells which transmit the ability to express delayed-type hypersensitivity and cell mediated immunity from sensitized donors to nonimmnune recipients. In this study, we determined the efficacy of TF as immunotherapy to treat experimental glioblastoma. We used TF obtained from immunized swine. We evaluated different doses of intratumoral TF (product of 4x10(6), 8x10(5) and 1.6x10(5) cells). The best dose (product of 4x10(6) cells) of TF was also combined with carmustine for experimental therapy in rats with C6 malignant glioma. Modifications in peripheral blood T lymphocyte counts ( CD2+, CD4+, CD8+ and NK) were evaluated by flow cytometry. Cytokine expression in the tumour was assessed by RT-PCR and apoptosis was evaluated using the sub G0 method. Intratumoral TF reduced significantly the tumour size, and increased CD2+, CD4+, CD8+ and NK cell counts, it also increased the percentage of apoptotic tumour cells and the percentage of tumour tissue expressing Th1 cytokines. We observed an additive antitumoral effect when TF was combined with chemotherapy. PMID- 16471321 TI - Antitumor resistance activation in mice: can the immunological memory cells enhance resistance? AB - Immunization of adult animals with the Ehrlich ascytic cancer cells homogenate three months prior to an experiment, did not affect either tumor transplantation or the progress of cancerogenesis induced by injection of 20-methylcholanthrene oil solution into the femoral muscle. All consequences of adult animal vaccination disappeared in 30-40 days following antigen administration. Quite different consequences were observed after immunization of the newborn mice. The same antigen (Ehrlich cancer cells homogenate) injected to newborn mice on days 1 and 3 after birth in a dose that failed to develop tolerance not only significantly increased the ascytic tumor transplantation threshold (by nearly 200 times for sarcoma 37 cells and by nearly 400 times for Ehrlich cancer cells) in adult animals but also led to almost 50% inhibition of cancerogenesis (induced by injection of 20-methylcholanthrene oil solution in the femoral muscle of mature mouse) after three and even after 12 months following immunization. The MTT-analysis did not reveal any noticeable differences in the number and activity of the cytotoxic lymphocytes in populations of splenocytes obtained from the intact mice (control) and from the adult animals which had been exposed to postnatal immunization (experiment).However, after a new vaccination such differences were found. In the populations of splenocytes obtained from control animals, the cytotoxic activity measured on day 10 after vaccination had increased 2.86-fold mainly at the expense of an increased number of effector cells. In the populations of splenocytes obtained from the experimental group of animals the activation was much greater (25.8-fold), being accomplished not only at the expense of an increased number of the effector cells, as observed in the control group, but also at the expense of their higher activity. The kinetic analysis of a mechanism of effector cells/target cells interaction has led to derive equations for estimation of the limiting rates of such interaction and of the equilibrium constants for interacting cells. Analysis of a generally accepted mechanism of the cytotoxic lymphocytes formation, with an account of the kinetic analysis data, has shown that a major reason of low antitumor resistance of animal organism is the negligible population of resting cells--the precursors of antitumor cytotoxic lymphocytes. Newborn mice vaccination does not produce any increase in the number of resting cells of the necessary type. This circumstance explains both, increase of the ascytic tumor transplantation threshold and increase of the resistance to 20-methylcholanthrene action in adulthood. Adult animal immunization does not possess such action. Analysis of the problem leads to the conclusion that the system of organism's antitumor resistance becomes effective only in those cases when, owing to antigen activation of resting cells, the concentration of cytotoxic lymphocytes rises to such an extent that the rate of tumor cell destruction becomes greater than the rate of target cells reproduction. PMID- 16471322 TI - WT1 expression level and clinical factors in multiple myeloma. AB - Although Wilm's Tuomor gene (WT1) was first identified as a tumor suppressor gene for Wilm's tumor, WT1 overexpression has been detected in different malignant cell types including leukemia. Increased expression of WT1 in acute leukemia is potentially used as a marker of minimal residual disease. However, the significance of the gene for multiple myeloma is still not clear. To determine the clinical relevance of WT1 expression in multiple myeloma, we examined the association of clinical parameters and WT1 expression in bone marrow for 17 newly diagnosed multiple myeloma patients. WT1 was assessed by real-time quantitative polymerase chain reaction (RQ-PCR) and calculated standardized WT1 expression level per 100 plasma cells in the bone marrow specimen as "corrected WT1". The expression of standardized WT1 and corrected WT1 in myeloma was 59 to 1,600 copies/microg RNA and 0.05 to 406.3 copies/microg RNA/100 plasma cells, respectively, lower than in leukemia. WT1 transcripts increased when clinical factors worsen, including the stage, amount of M protein, Hb, platelet count, blood urea nitrogen (BUN), creatinine, serum alkaline phosphatase (ALP), calcium, beta2-microglobulin, thymidine kinase activity (TK), and C-reactive protein (CRP). In conclusion, the expression level of WT1 could be an additional marker to the standard parameters considered in risk assessment for multiple myeloma. PMID- 16471323 TI - Over expression of aromatase protein is highly related to MMPs levels in human breast carcinomas. AB - Estrogens can play a critical role in the development of breast cancer. Aromatase which catalyzes the formation of aromatic C18 estrogens from C19 androgens is regarded to be responsible for the cancer local production of estrogen. Studies not only from aromatase transfected breast cancer cells, but also transgenic mouse, which overexpressed aromatase, demonstrated that in situ produced estrogen plays more important roles than circulating estrogens in breast tumor promotion and progression. Matrix-metalloproteinases (MMPs) have been implicated in the proeolytic process, which play important roles in the aggressiveness of cancer cells including invasion of adjacent tissue and metastasis to distant sites. Expression of MMP2 and 9 may be stimulated by estrogens in hormonal dependent breast cancers, since tumor aromatase can stimulate breast cancer growth and progression in both an autocrine and a paracrine manner. Theoretically aromatase overexpression, that causes relatively high estrogen concentration in situ, may be positively related to MMP2 and 9 expression, indicate worse prognosis in breast cancers, and maybe insensitive to tamoxifen therapy. In the present study, we studied the expression of aromatase activity and MMPs in human breast carcinoma both in vitro and in vivo. In human breast carcinoma cell lines including MCF-7, MDA-MB-231 and MDA-MB-435, the expression of aromatase levels both in mRNA and protein activity was related to MMP2 and MMP9. In humam breast cancer samples, we demonstrated that aromatase expressions were strongly associated with MMP2 and MMP9 levels. It was interesting to observe that the positive relationship was only present in the ER and/or PR positive patients. This may indicate that both MMP2 and MMP9 were up regulated by estrogen produced by aromatase through ER. So in endocrine therapy, either blocking the ER by tamoxifen or inhibiting the aromatase by aromatase inhibitors for example letrozole, may both inhibit tumor growth and lower the metastatic potential especially in ER positive breast cancer patients by means of down-regulation of MMP2/9. PMID- 16471324 TI - Evaluation of DNA content in preneoplastic changes of mouse urinary bladder induced by N-butyl-N-(4-hydroxybutyl) nitrosamine. AB - Using image cytometry analysis we analysed the deoxyribonucleic acid content of preneoplastic lesions induced in C3H/He female mice. Female mice (n = 30) were given 0.05% BBN in their drinking water for 4, 8 and 12 weeks, and were then euthanized. At the 4th week 90% hyperplasias, 90% dysplasias and 10% papillary tumours developed selectively in the bladder, after 8 weeks mice developed 80% hyperplasias and 80% dysplasias and 12 weeks we identified 70% hyperplasias, 70% dysplasias, 10% papillary tumours and 20% squamous metaplasia. Among the ones who developed simple hyperplasia, 58.3% were diploid and 41.7% were aneuploid. In the dysplasia lesions 29.2% were diploid and 70.8% were aneuploid (p = 0.041). All papillary tumours were aneuploid and all epidermoid metaplasias were diploid. These results suggest that aneuploidization, DNA content alteration of preneoplastic lesions, might be considered as a prognostic factor. PMID- 16471325 TI - Effect of incadronate on proliferation of mesenchymal tumor cells with or without activated Ras mutation. AB - The present investigation examined the effect of bisphosphonates on six mesenchymal tumor cell lines and the mechanisms of inhibition of tumor cell proliferation. HT-1080, a fibrosarcoma cell line that exhibits increased Ras activity due to a mutation of the Ras gene, demonstrated significantly reduced tumor cell proliferation upon treatment with incadronate. The other cell lines, however, which lack mutation of the Ras gene, showed no influence upon treatment with incadronate. Autoradiography demonstrated no difference in the uptake of 3H labelled incadronate between susceptible and unaffected cells. The anti proliferation of HT-1080 was reversed by the addition of geranylgeranyl pyrophosphate. Etidronate exhibited no influence on all tested cell lines. On the basis of these data, we hypothesized that incadronate inhibits the mevalonate pathway and blocks oncogenic Ras signaling. In an effort to confirm this hypothesis, the influence of incadronate on an oncogenic Ras transfected BALB/3T3 cell line (Bhas 42) and a parental BALB/3T3 cell line were compared. The parental BALB/3T3 cells showed slight inhibition upon treatment with incadronate, however, the proliferation of Bhas 42 cells was significantly reduced. These results suggest that incadronate suppresses oncogenic Ras-activated mesenchymal tumors through the inhibition of Ras signaling pathways. PMID- 16471326 TI - High concentration of deoxycholic acid abrogates in vitro transformation of IEC6 intestinal cells by azoxymethane. AB - In this study, we designed an in vitro azoxymethane (AOM)-induced carcinogenesis model and analyzed the effect of deoxycholic acid (DCA) on growth, apoptosis, genotoxicity, and transformation of IEC6 intestinal cells. CYP2E1 production was confirmed in IEC6 cells. The growth of IEC6 cells was enhanced by DCA (100 microg/ml). However, IEC6 cells treated with DCA (200 microg/ml) were inhibited and disappeared at 48 hrs after treatment. Apoptotic cells increased 11.2 times by treatment with DCA (200 microg/ml) as compared to cells with no treatment. DNA injury detected by comet assay was found in cells treated with AOM, but not in cells treated with DCA (100 microg/ml) and AOM. The number of colony formation in soft agar increased by AOM treatment. However, the number of foci treated with DCA (100 microg/ml) plus AOM was 69% that of cells treated with AOM alone. Two out of the 6 mice subcutaneously injected with AOM-treated IEC6 cells showed tumorigenesis, whereas IEC6 cells treated with DCA (100 microg/ml) plus AOM or DCA (100 microg/ml) alone did not form any tumor. Reduced protein expression of MLH1, Bcl-2 was detected in IEC6 cells treated with DCA (100 microg/ml). Production of Bax, pJNK, TGF-beta, TGFBRI, TGFBRII, and beta-catenin were higher in IEC6 cells treated with DCA (100 microg/ml) than that in cells with no treatment. These results suggest that high-dose DCA induced apoptosis and inhibited AOM-induced in vitro transformation of IEC6 cells. PMID- 16471327 TI - C-erb-2 gene amplification and chromosomal anomalies in bladder cancer: preliminary results. AB - Studies recently suggested that different genetic factors are involved in the development and progression of bladder cancer. In this study, 30 consecutive patients affected by bladder neoplasm were evaluated in order to analyze the frequency of c-erb-2 gene amplification and chromosome 7, 9, 17 aneusomy using fluorescence in situ hybridization (FISH) technique. C-erb-2 gene amplification, chromosome 17 gain and aneusomy were respectively observed in 3.7% (1/27), in 47% (12/27) and in 74% (20/27) of examined tumors. Moreover, chromosome 7 and 9 aneusomy were detected in 74% (20/27) and in 72% (16/27) of specimens. A statistically significant correlation was observed between chromosome 17 aneusomy and tumor stage and grade (r: 0.642, p = 0.0001; r: 0.385, p = 0.04, respectively). In conclusion, we observed a low incidence of C-erb-2 gene amplification, while chromosome 17 aneusomy was confirmed as a marker of advanced and aggressive bladder cancer. PMID- 16471328 TI - Human telomerase reverse transcriptase expression in oral carcinogenesis--a preliminary report. AB - Human telomerase reverse transcriptase (hTERT), the catalytic subunit of telomerase, is strongly associated with telomerase activity implicated in cellular immortalization and tumorigenesis. In situ detection of hTERT will aid in determining the localization of telomerase positive cells. The aim of this study was to detect hTERT protein expression in multistep oral carcinogenesis using paraffin embedded tissue samples, and to study the relationship of hTERT expression with different histological stages in oral carcinogenesis. Normal (n = 4), hyperplastic (n = 4), dysplastic (n = 4) and neoplastic (n = 10) oral epithelia representing different histological stages in oral carcinogenesis were included in the study. hTERT protein detection was done by immunohistochemistry (IHC) technique. Nuclear staining intensities were noted and the hTERT-labelling index was determined. Dysplastic and neoplastic oral epithelia showed an increased percentage of hTERT positive cells (Grade 4: > 50% positive staining nuclei) with intense staining in the basal, parabasal and superficial layers of the epithelia, unlike normal oral mucosa which showed intense staining only in the basal and parabasal cell layers, which are the normal proliferative progenitor compartments. hTERT protein expression was elevated with the corresponding advancement of the histological stages of oral carcinogenesis, from normal to hyperplasia to dysplasia to carcinoma. There seems to be an upregulation of hTERT protein expression during the progression of oral cancer, therefore, this may indicate the feasibility of IHC detection of hTERT protein in oral carcinogenesis as a potential diagnostic or prognostic marker. PMID- 16471330 TI - [Field investigation on Salix psammophila plant morphology and airflow structure]. AB - A field investigation in the flat sands of southern Maowusu sandy land showed that artificially cultivated single line Salix psammophila could accumulate sand because the plant decreased the windward and leeward wind velocity. There was a significant correlation (R = 0. 696) between accumulated sand volume (V2) and plant volume (V1). When the wind velocity at 4 m height of single-line S. psammophila was 6 m x s(-1), the wind velocity decreased at 3H windward, increased at 2H windward, and then steeply decreased leeward, reached the lowest value at 1H leeward and gradually recovered to the open field velocity. The protection distance of single-line S. psammophila was about 17 H', and the effective protection distance was about 13 H'. Single-line S. psamnmophila had little effect on the wind velocity above plant height. PMID- 16471329 TI - A case of esophageal small cell carcinoma with multiple liver metastases responding to chemotherapy with irinotecan plus cisplatin. AB - We report a case of small cell esophageal carcinoma (SCEC) with multiple liver metastases treated with some success by chemotherapy with irinotecan (CPT-11) plus cisplatin (CDDP). Radiologic and endoscopic examination of a 75-year-old man with multiple liver tumors disclosed a 4.0-cm type 2 tumor in the middle third of the esophagus. An endoscopically obtained biopsy specimen was diagnosed as undifferentiated small cell carcinoma. Multiple liver metastases were confirmed but lymph node metastases and distant metastases other than those in the liver were not detected. After six courses of chemotherapy with CPT-11 plus CDDP, the primary lesion showed complete response and liver metastases showed partial response. However, because all lesions almost immediately relapsed or progressed, arterial infusion chemotherapy for liver metastases and radiation for the primary lesion were given as second-line treatment. The primary lesion showed complete response with radiation. Arterial infusion chemotherapy prevented the progression of liver metastases once, but the patient died of liver failure at last. No distant lesions including metastatic lymph nodes were confirmed over the course of his illness, and the patient survived for a year after first diagnosis. Although the prognosis of SCEC is quite unfavorable due to highly aggressive behavior, a better prognosis is possible with effective chemotherapy and second line treatment is important in improving prognosis. PMID- 16471331 TI - [Physicomechanical properties of plantation poplar wood under different eco environments]. AB - The measurement of the physicomechanical properties of 9 plantation poplar I-69 (Populus deltoides CV. 1-69/55) woods under fertilization and seasonal flooding showed that compared with no fertilization, fertilization increased the ring width, static modulus of elasticity, and compressive strength parallel to the grain by 0.99 % (P > 0.05), 2.73% (P < 0.05) and 1.06% (P < 0.05), while decreased the air-dry radial, tangential, volumetric shrinkage, oven-dry radial, tangential, volumetric shrinkage, basic density, MOR, and dynamic modulus of elasticity by 4.2%, 7.7%, 6.6%, 15.6%, 6.3%, 11.1%, 2%, 1.79% and 8.13%, respectively. Seasonal flooding decreased the basic density, air-dry radial, tangential, volumetric shrinkage, oven-dry radial, tangential, volumetric shrinkage, MOR, MOE, and compressive strength parallel to the grain by 5.5%, 11.1%, 9.2%, 9.6%, 16.7%, 10.9%, 8.5%, 24.29%, 18.18% and 16.81%, respectively, in comparing with non-flooding. PMID- 16471332 TI - [Interspecific association between understory species in a southern highland plantation]. AB - Based upon 2 x 2 contingency table, chi2 test and association coefficient were used to determine the interspecific association between understory species in a southern highland plantation, and to analyze the restoration degree and the stability of southern highland vegetations originated from plantation. The Qianyanzhou in Taihe County of Jiangxi Province, a typical sample of southern highland plantation, was chosen to make the study. The results showed that both in shrub layer and in herb layer, species pair with chi2 reaching significant level (P <0.05) was few in number. In shrub layer, 12 species pairs' association was highly significant (P < 0.01), 19 pairs' was significant (P < 0.05), and other 200 pairs' was nonsignificant, while in herb layer, 11 pairs' was highly significant, 11 pairs' was significant and other 83 pairs' was nonsignificant. According to interspecific association and correlation, shrub layer was divided into two species groups: Group I . Adinandra bockiana, Syzygiumn grijsii, Vaccinium bracteatunm, Ilex aculeolata, Smilax ferox, Eurya muricata and Group II . Lespedeza davidii, Serissa serissoides, Vitex negundo var. cannabifolia. Many species in Group I had a significantly negative association with the species in Group II, and dominant species always played a key role in the relationships among species. The three dominant species in herb layer, Wooduardia japonica, Dryopteris atrata and Adiantun flabellulaturn, had a highly significant positive correlation between each other, and moreover, had a significant or highly significant positive association with many other herbaceous species. Similarily, dominant species in shrub layer played a key role on the interspecific association in the two species groups. The ratios of positive and negative association indicating the species compositions of the two layers were fluctuating, which was 125/106 in shrub layer and 42/63 in herb layer. Several shortcomings of interspecific association method were pointed out, with some proposals put forward. PMID- 16471333 TI - [Natural vegetation restoration and soil nutrient dynamics of abandoned farmlands in forest-steppe zone on Loess Plateau]. AB - To understand the relationship between plant community succession and soil nutrient dynamics is crucial in intervening vegetation succession. This paper reported the results from a study carried out in a forest-steppe zone on the Loess Plateau, with emphasis on the vegetation characteristics and soil nutrient dynamics duringvegetation restoration on abandoned farmlands of this area. The results showed that under zonal habitat conditions, natural vegetation succession would turn toward the original vegetation communities, but the expected shrub or trees communities didn't occur after 40 approximately 50 years, and Stipa bungeana, Artemisia sacrorum, Bothriochloa ischaemun, Stipa grandis and Lespedeza davurica communities were still the widely distributed communities. Vegetation restoration had a significant effect on soil nutrient dynamics. In general, soil nutrient contents increased with restoration time, and extremely significant changes occurred in soil organic matter, total N, available N, and available K (P< 0.001),while only significant changes in available P (0.05 < P < 0.01) and no significant change in total P (P > 0.05). Moreover, the changes were also found in soil profile. Statistical analysis showed that from surface layer (0 approximately 20 cm) to deeper layer (to 60 cm), extremely significant changes occurred in soil organic matter, total N, available N, available K and available P (P < 0.001), and significant change in total P (P< 0.05). Soil nutrients intended to accumulate in surface layer. Further linear correlation analysis showed that soil organic matter, total N, available N and available K were significantly correlated each other ( P< 0.001), but didn't show correlation with soil total P and available P. PMID- 16471334 TI - [Decomposition of Triploid populus tomentosa fine root and Lolium multiflorum grass root in a composite ecosystem and their nutrient dynamics]. AB - This paper studied the root decomposition and its nutrient dynamics of Triploid populus tomentosa and Lolium multiflorum in a plantation converted from farmland at Tianquan County of Sichuan Province. The results showed that the weight loss rate of fine root and grass root showed an exponential relationship with time, with an annual loss of 76.17%, 69.80%, 73.44% and 79.53% for fine roots of 0 approximately 1, 1 approximately 2 and 0 approximately 2 mm in diameter and grass root, respectively. During the decomposition of fine roots, their N and Ca contents increased, while P, K and Mg contents were in adverse. The nutrient contents of grass root during its decomposition had an irregular variation. The remaining percentage of fine root P, K and Mg had the same trend as root weight loss during its decomposition, while that of N and Ca had a slight decrease. For grass root, its N, P, K, Ca and Mg contents declined rapidly at the initial stage of decomposition, and then decreased slowly with irregular deposition rate. PMID- 16471335 TI - [Transpiration of Choerospondias axillaris in agro-forestrial system and its affecting factors]. AB - Measurement of transpiration is essential to assess plant water use efficiency. Applying Grainer method, this paper measured the sap flow of Choerospondias axillaries in an agro-forestrial system, aimed to evaluate the effects of intercropping and pruning on the diurnal variation of transpiration, and to relate the transpiration rate with climatic factors. The results showed that the diurnal variation of Choerospondias arillaries transpiration rate appeared in parabola, low in the morning and evening, and high at noon. The transpiration rate was closely related to leaf stomatal conductivity and soil water potential, especially the water potential in 100 cm soil depth (R = 0.737). The transpiration rate of Choerospondias axillaries was increased by about 40% approximately 160% in agro-forestrial system through the changes in regional environment and in the deep soil water use by tree. Correlation analysis and multi-factor successive regression analysis indicated that the transpiration was controlled by ray radiation intensity, air temperature and ground temperature, followed by the difference between saturated and actual vapor pressure and the wind speed. A statistical model for calculating the sap flow rate by micrometeorological factors was also provided. PMID- 16471336 TI - [Dynamics of soil properties under secondary succession forest communities in Mt. Jinyun]. AB - Mt. Jinyun is located in the north suburb of Chongqing, 30 km away from the city center. It is rich in forest plants, an epitome of forests in north tropical areas of China. Under anthropocentric disturbance, there still exist large numbers of succession communities, and the process of successive development follows the way of shrub-grassland (X1)-->coniferous forest (X2)-->coniferous broad leaved mixed forest (X3)-->evergreen broad-leaved forest (X4). By now, soil and water conservation is very important in the Three Gorges area of Yangtze River, and the investigation on the secondary succession of the forests could help to realize the changes of the forests and soils under anthropocentric disturbance, and supply information on the protection of natural forests and the artificial reforestation of this area. In this paper, some typical and representative plant communities in different succession stages were selected to study the plant composition and type and the soil properties, with species diversity indices and canopy density investigated in many standard squares and soil physical and chemical characteristics analyzed. The results showed that there were obvious variations of soil properties with time. As the plant community developed from primary stage to climax, the contents of soil organic matter, total N, and available N and K increased in order of X1 < X2 < X3 < X4, soil pH changed from 5.23 (X1) to 4.06 (X4), soil base saturation varied from 58.3% (X1) to 37.7% (X4), and soil CEC increased with the succession. It was suggested that an intense soil acid leaching was occurred in Mt. Jinyun. The contents of soil organic matter and total N in different layers showed a trend of A>B>C, e. g., soil total nitrogen in evergreen broad leaved forest was 2.31(A), 0.66(B) and 0.12(C)g x kg(-1). Gray analysis was used to study the relationships of soil properties between the climax community and other three succession communities. The relation coefficient was 0.461 0 (X3), 0.586 2(X2) and 0.6821 (X1), respectively, indicating that soil nutrients were accumulated as the forest succession community progressed. The plant arbor species followed the sequence of 0 (X1) < 7.5 (X2) < 9.0 (X3) < 12.8 (X4), and the canopy density ranked as the same way. Plant community could affect soil nutrient reserves significantly. Multivariate variance analysis showed that soil properties varied significantly among different seasons, but this variation had no impact on the community replacement and soil development during the chronosequence of community succession. The variation of soil properties adapted well to each successive community. PMID- 16471337 TI - [Construction of information management-based virtual forest landscape and its application]. AB - Based on the analysis of the contents and technical characteristics of different scale forest visualization modeling, this paper brought forward the principles and technical systems of constructing an information management-based virtual forest landscape. With the combination of process modeling and tree geometric structure description, a software method of interactively and parameterized tree modeling was developed, and the corresponding renderings and geometrical elements simplification algorithms were delineated to speed up rendering run-timely. As a pilot study, the geometrical model bases associated with the typical tree categories in Zhangpu County of Fujian Province, southeast China were established as template files. A Virtual Forest Management System prototype was developed with GIS component (ArcObject), OpenGL graphics environment, and Visual C++ language, based on forest inventory and remote sensing data. The prototype could be used for roaming between 2D and 3D, information query and analysis, and virtual and interactive forest growth simulation, and its reality and accuracy could meet the needs of forest resource management. Some typical interfaces of the system and the illustrative scene cross-sections of simulated masson pine growth under conditions of competition and thinning were listed. PMID- 16471338 TI - [Spatial variance characters of urban synthesis pattern indices at different scales]. AB - Scale holds the key to understand pattern-process interactions, and indeed, becomes one of the corner-stone concepts in landscape ecology. Geographic Information System and remote sensing techniques provide an effective tool to characterize the spatial pattern and spatial heterogeneity at different scales. As an example, these techniques are applied to analyze the urban landscape diversity index, contagion index and fractal dimension on the SPOT remote sensing images at four scales. This paper modeled the semivariogram of these three landscape indices at different scales, and the results indicated that the spatial variance characters of diversity index, contagion index and fractal dimension were similar at different scales, which was spatial dependence. The spatial dependence was showed at each scale, the smaller the scale, the stronger the spatial dependence. With the scale reduced, more details of spatial variance were discovered. The contribution of spatial autocorrelation of these three indices to total spatial variance increased gradually, but when the scale was quite small, spatial variance analysis would destroy the interior structure of landscape system. The semivariogram models of different landscape indices were very different at the same scale, illuminating that these models were incomparable at different scales. According to above analyses and based on the study of urban land use structure, 1 km extent was the more perfect scale for studying the spatial variance of urban landscape pattern in Shanghai. The spatial variance of landscape indices had the character of scale-dependence, and was a function of scale. The results differed at different scales we chose, and thus, the influence of scales on pattern could not be neglected in the research of landscape ecology. The changes of these three landscape indices displayed the regularity of urban spatial structure at different scales, i. e., they were complicated and no regularity at small scale, polycentric at moderate scale, and circle-zoning at big scale. PMID- 16471339 TI - [Impact of urbanization on pedodiversity in Suzhou area]. AB - Based on the TM remote sensing information of 1984, 1995, 2000 and 2003, this paper quantitatively analyzed the spatial-temporal dynamic changes of pedodiversity in Suzhou area under quickly growing urbanization. The results showed that in this area, clay loamy typic-hapli-stagnic anthrosol and fine sand clay loamy typic-hapli-stagnic anthrosol were the predominant soil types, but their distribution area decreased 5.11% and 3.14%, respectively, in latest 20 years. Jinchang, Pingjiang and Canglang Districts were the focuses of urbanization in Suzhou area, where pedodiversity changed furiously. During 1984 approximately 2003, more than 90% of the clay loamy typic-hapli-stagni gleyosol in Pingjiang District and sandy calcaric-mottlic-udi-orthic primosol in Taicang City were almost disappeared, and changed into residential area. Statistical analysis showed that urbanization had a significant impact on pedodiversity, and was the prime driving force on the pedodiversity in Suzhou area. PMID- 16471340 TI - [Spatial-temporal dynamics of landscape fragmentation in North Shannxi Loess Plateau]. AB - The study of landscape fragmentation is an important means to reveal regional ecological processes. By using the remote sensing data of 1987, 1997 and 2002, and the GIS tools, some representative quantitative indices of the landscape spatial structure and landscape fragmentation in North Shannxi Loess Plateau were calculated, aimed to investigate the relationships between landscape ecological structure and landscape pattern in this area. The results showed that cropland contributed most to the extent of landscape fragmentation, but unused land lacked obvious relationship with it. To get large-area images accurately and promptly is very important for monitoring ecological process in regional scale. Moderate Resolution Imaging Spectroradiometer (MODIS) images in 2003 were used to derive monthly landscape fragmentation information calculated with Local Index of Spatial Associations (LISA) based on Spatial Statistical Analysis (SSA). The analysis of the fragmented landscape types showed that human disturbance had a main contribution to the seasonal landscape fragmentation. Furthermore, with the increasing percentages of cropland and unused land, landscape became more fragmented, while forestland coverage had inverse effects on the fragmentation. PMID- 16471341 TI - [Effects of N fertilization on winter wheat grain yield and its crude protein content and apparent N losses]. AB - Field experiments were conducted at four sites in 2002 to study the effects of N fertilization on winter wheat yield and its crude protein content and apparent N losses. The results showed that at these sites, the N application rate for maximum grain yield was 0, 0, 79 and 118 kg x hm(-2), and that for maximum grain crude protein content was 122, 100, 127 and 174 kg x hm(-2), respectively. The residual nitrate-N content in 0 approximately 90 cm soil layer after harvest was linearly increased, while the apparent N losses during wheat growth season was exponentially increased with increasing N application rate. Under the N application rate for maximum yield, soil residual nitrate-N content ranged from 86 to 115 kg x hm(-2) and apparent N losses ranged from 2 to 32 kg x hm(-2), which were much lower than the residual nitrate-N content (106 approximately 168 kg x hm(-2))and apparent N losses(14 approximately 56 kg x hm(-2)) when the maximum grain crude protein content was achieved. The results showed that the maximum wheat grain yield and the environment effects of N fertilization could be harmonized by an optimized N application rate, but, if considering the importance of wheat grain protein, more research in the future should be needed. PMID- 16471342 TI - [Dynamics of soil enzyme activity and nutrient content in intercropped cotton rhizosphere and non-rhizosphere]. AB - The study with high yield cotton-wheat double cropping system showed that soil urease, invertase, protease and catalase activities in intercropped cotton field had the same changing trends with those in mono-cultured cotton field, but were significantly higher in intercropped than in mono-cultured cotton rhizosphere and non-rhizosphere at all development stages of cotton. During the intergrowth period of wheat and cotton, soil nutrient contents in intercropped cotton rhizosphere and non-rhizosphere were lower than or had little difference with those in mono-cultured cotton rhizosphere and non-rhizosphere, but became significantly higher after wheat harvested. The changing trends of soil nutrient contents in intercropped cotton field had little difference from those in mono cultured cotton field, but the nutrient absorption peak appeared late. The soil enzyme activities and nutrient contents were generally higher in rhizosphere than in non-rhizosphere of both intercropped and mono-cultured cotton. Soil nutrient contents had significant (P < 0.05, n = 32) or very significant (P < 0.01, n = 32) correlation with the activities of soil urease, invertase and protease, but had little correlation with soil catalase activity. PMID- 16471343 TI - [Distribution of C and N in different particle fractions of meadow brown soil in lower reach of Liaohe River Plain]. AB - The study of meadow brown soil with different reclamation history in the lower reach of Liaohe River Plain showed that in 0 approximately 20 and 20 approximately 40 cm soil layers, <0.002 mm fraction had the highest content of organic carbon, followed by < 0.02 and < 0.2 mm fractions. The distribution pattern of total nitrogen was similar to that of organic carbon. There was no obvious correlation between the distribution of organic carbon and total nitrogen in different soil particle fractions and the history of reclamation, but the correlation between organic carbon and total nitrogen in different soil particle fractions was highly significant, with the correlation coefficients 0.74, 0.94 and 0.91 in fractions< 0.2, < 0.02 and < 0.002 mm, respectively. PMID- 16471344 TI - [Spatial variability of soil phosphorus in field scale]. AB - In this paper, the spatial variability of soil total P and Olsen-P at the depths of 0 approximately 10 and 10 approximately 20 cm in a field-scale was examined by using traditional statistics combined with geostatistics methods. A sampling grid of 30 m x 42 m including 49 pairs of soil sampling points was established in the Shenyang Experimental Station of Ecology, Chinese Academy of Sciences. The results showed that the variance coefficients were much higher for Olsen-P (46.56% approximately 56.42%) than for total P (11.68% approximately 13.33%). Both total P and Olsen-P in the two soil depths had strongly spatial structures and similar spatial correlation ranges. The parameters derived from best-fitted models showed that the spatial variability of both total P and Olsen-P was mainly affected by structural factors, with C/(C0 + C) being higher than 66% for all the variables. The kriging contour maps showed that both total P and Olsen-P in the two soil depths had similar spatial distribution patterns. A more precise sampling scheme could be made based on the spatial distribution pattern of test soil properties combined with traditional variance coefficients. To recognize the strong variability of soil Olsen-P in field-scale is of significance for better understanding the P cycling in farm ecosystems and for precise agriculture. PMID- 16471345 TI - [Main affecting factors of soil wind erosion under different land use patterns--a case study in Wuchuan County, Inner Mongolia]. AB - Field investigation, laboratory analysis and wind tunnel simulation showed that in Wuchuan County of Inner Mongolia, low precipitation, frequent and high wind velocity, coarse soil texture, and thawing and freezing were the main causes of soil wind erosion happened very easily in spring. In late winter and early spring, the vegetation coverage was in order of shrub-land>natural grassland>rainfed farmland, and thus, increasing the surface cover of rainfed farmland should be an urgent need to control the wind erosion in Wuchuan County. The soil wind erosion rate decreased exponentially with increasing soil moisture content, and 6% soil moisture content was a turning point from severe to light. The topsoil moisture content under different land use patterns was in order of natural grassland> rainfed farmland >shrub-land. With increasing wind velocity, soil wind erosion rate increased by power function, and 18 m x s(-1) wind velocity was a switching point to aggravate the wind erosion. PMID- 16471346 TI - [Effects of rhizosphere CO2 concentration on potato growth]. AB - With the method of areoponics, this paper established a rhizosphere gas environment research system to study the effects of different rhizosphere CO2 concentration on the growth of potato (Solanum tuberosum) plants. The results showed that in treatments 380 and 380 approximately 920 micromol CO2 x mol(-1), the plant height, stem diameter, leaf area, root length, and individuals of stolon and mini-tuber had a similar variation trend with time, and presented two distinct phases. The plants grew vigorously, and their tuber yield increased markedly, compared with treatment 3,600 micromol CO2 x mol(-1), which indicated that 380 approximately 920 micromol CO2 x mol(-1) was the appropriate CO2 concentration in the root zone to promote potato growth, while overmuch rhizosphere CO2 would markedly inhibit the growth and development of potato plants. Treatment 3,600 micromol CO2 x mol(-1) and sand culture had the similar stem diameter, leaf area, and individuals of stolon and mini-tuber. The plants grew short and weak, and tuber yield was lower. PMID- 16471347 TI - [Quantitative analysis on the relationships between cotton fiber quality and meteorological factors]. AB - With the method of planting cotton by stages at the sites from Yangtze valley to north Xinjiang, this paper studied the effects of air temperature and sunlight on cotton fiber quality in 2002 approximately 2003. The results showed that with increasing latitude, fiber length increased, while micronaire value and soluble sugar content (SSC) decreased. An ecological model describing the relationships of fiber strength, micronaire value and SSC with air temperature and sunlight was built to determine the optimal range and critical value of air temperature during boll development period. If a high quality fiber (length > or = 27 mm) was targeted, the daily mean temperature (DMT) and minimum temperature (Tmin) during this period should be 21.3 approximately 29.7 and 10.7 approximately 21.3 degrees C, respectively, and the daily temperature difference (DTD) should not be > 15.2 degrees C. Fiber length was the longest when the DMT was 25.4 degrees C. If the lowest fiber length was > or = 25 mm, DMT and Tmin should not be lower than 15.5 and 10.7 degrees C, respectively, and DTD should not be over 17.9 degrees C during boll development period. PMID- 16471348 TI - [Analysis on the role and endowment of virtual water in agricultural ecosystem]. AB - Water is one of the most important components of agricultural ecosystem, and also a source of food security, which plays an essential role in ensuring sustainable food resources for a growing population. In this study, the policy relevancy of virtual water was addressed, with food production, alternative trend of import, and product condition analyzed. The results showed that an average of 10.2 x 10(9) m3 water and 27.7 x 10(6) hm2 land was transferred by imported wheat, while an average of 0.3 x 10(9) m3 water and 1.6 x 10(6) hm2 land was transferred by imported maize. The sum of virtual water equivalent was 10.5 x 10(9) m3. It's of significance to alleviate the pressure of water stress in agriculture and arable land, help to resolve water predication of food production by food imports, and compensate water endowment. PMID- 16471349 TI - [Root growth characteristics of soybean and Faba bean at their seedling stage]. AB - Soybean and faba bean are the two main legumes widely intercropped with cereals (e. g., wheat or maize) in western China, but have significantly different performance in intercropping system. Compared to soybean, faba bean has a stronger competitive ability. Considering that underground interspecific interaction makes a significant contribution to the advantage of intercropping system, a pot experiment was conducted to compare the differences in root growth and morphological parameters between soybean and faba bean at their seedling stage. A PVC tube, with 10 cm in diameter and 50 cm in depth, was filled with quartz and used to grow the crops. Root systems were scanned by a scanner, and analyzed by WinRHIZO on days 14, 21, 28, 35 and 42 after seedling emergence. The results indicated that on the 42nd day after seedling emergence, the root surface area of faba bean was 2.61 times larger than that of soybean. Soybean roots distributed in the whole layer of quartz on the 14th day, but most faba bean roots only distributed in its top layer. Soybean roots were thinner, and most of them were 0.25 approximately 1.0 mm in diameter, while faba bean roots were thicker, and their diameter was mostly between 0.5 approximately 1.5 mm. All of these facts resulted in a higher absorption capacity and nutrient use efficiency of faba bean, which could be one of the reasons that faba bean had a stronger competitive capacity than soybean. PMID- 16471350 TI - [Effects of weak light and high temperature stress after anthesis on flag leaf chlorophyll fluorescence and grain fill of wheat]. AB - A field experiment with wheat showed that after anthesis, the photosystem II (PS II), photochemical efficiency (Fv/Fm) and photosynthetic rate (Pn) of flag leaf decreased substantially, its actual quantum yield of PS II electron transport (phiPS II) and photochemical quenching co-efficient (qP) declined slightly, while its non-photochemical quenching (NPQ) increased slightly three days after shading, compared with the control. Three days after high temperature treatment, the Fv/Fm, Pn, phiPS II and qP decreased significantly, while the NPQ increased slightly. During the course of recovery after the stresses, the parameters fluorescence and Pn which were stressed by shading and high temperature at prophase recovered slightly, while those stressed at metaphase decreased all along, indicating that the damage at prophase was reversible, but the stress at metaphase accelerated the caducity of flag leaves. The grain-filling processes of different treatments were modeled with Logistic equation, which showed that the decline of per kernel weight was mainly caused by the decrease of mean filling rate, maximal filling rate, and slow-increase period filling rate. Filling duration, occurrence time of maximal filling rate, duration of slight-increase period, and slight-increase period filling rate were less affected by weak light and high temperature. PMID- 16471351 TI - [Population dynamics of pests and their enemies in different cultivated rice fields]. AB - Through a whole year field investigation, this paper analyzed the population dynamics of pests and their enemies in organic and normally cultivated rice fields. The results indicated that the population of main pests varied more lightly in organic than in normally cultivated rice field, while subordinated pest species had little difference between these two fields. The individuals of enemies were more in organic than in normally cultivated rice field. It is suggested that the pest-control effect of enemies could be restored and became stronger when no chemical pesticides were applied. PMID- 16471352 TI - [Spatial distribution pattern and space dependence of Contarinia sp. and Erigonidium graminicola in jujube orchard]. AB - With the principles and methodologies of geostatistics, this paper studied the spatial structure and space dependence of Contarinia sp. and its natural enemy Erigonidium graminicola in a jujube orchard at different date. The results indicated that the semivariogram of Contarinia sp. on 8 June, 27 August and 19 October could be described by spherical model, showing an aggregated spatial distribution with the ranges of spatial dependence being 26.3, 7.8 and 22.0 m, and the degrees of spatial dependence being 88.05%, 85.77% and 87.58%, respectively. The semivariogram of Erigonidium graminicola could be also described by spherical model, showing an aggregated distribution with the ranges of spatial dependence being 28.3, 22.5 and 22.2 m, and the degrees of spatial dependence being 90.09%, 87.96% and 85.72%, respectively. It is suggested that Erigonidium graminicola is the dominant natural enemy against Contarinia sp. population in the examined jujube orchard. PMID- 16471353 TI - [Community succession patterns of pests and natural enemies in jujube orchard ecosystem]. AB - Through investigation and by the methods of optimal partitioning, principal component analysis, and one-or two-dimensional ordination, this paper analyzed the main groups and succession trends of the pest and natural enemy communities at a jujube orchard of Taigu area. The results showed that the communities had more obvious dominant factors and temporal succession pattern. The first three principal components of the communities had a bigger variation, with an obvious follow effects. The community succession was temporally divided into four stages, i. e., in the periods of pre-budding, leaf expanding and blooming, fruiting, and maturing. The dominant species of pests and their natural enemies were demonstrated. PMID- 16471354 TI - [Effects of Meloidogyne incognita initial inoculation density on ginger growth]. AB - The study with potted ginger showed that Meloidogyne incognita infection could reduce the plant height, stem diameter, quantity of shoots, and fresh weight of stem, leaf and root, and finally, reduce ginger yield. With the enhancement of M. incognita initial inoculation density, the growth of ginger was damaged more severely, e. g., under inoculating 50, 100 and 200 M. incognita eggs per 100 g soil, ginger yield decreased by 27.91%, 37.73% and 42.14%, respectively. The reproduction rate of M. incognita was also affected by the initial inoculation density, the lower the initial inoculation density, the higher the reproduction rate. When the inoculation density reached 746.20 eggs per 100 g soil, the reproduction rate was 1. PMID- 16471355 TI - [Effects of avermectin B1a on soil microorganism and earthworm (Eisenia fetida)]. AB - Avermectins are the pesticides widely applied in agriculture. This paper studied the effects of different concentration avermectin B1a (AVM B1a) on soil bacteria and fungi, soil respiration, and soil earthworm (Eisenia fetida). The results showed that in 4 test soils, bacteria were significantly inhibited when the AVM B1a concentration was higher than 83.3 mg x kg(-1), while fungi were less impaired. Soil respiration was also inhibited by high concentration AVM B1a, which differed with soil types. The half lethal dosage (LD50) of AVM B1a to soil earthworm was estimated as 4.63 microg x cm(-2) in filter paper contact test, and as 24.13 and 17.06 mg x kg(-1), respectively after treated 7 and 14 days in artificial soil. PMID- 16471356 TI - [Labeling of biocontrol agents ZJY-1 and ZJY-116 gfp gene and its ecological adaptability in cucumber rhizosphere]. AB - The recombined plasmid pRP22-GFP contained with gfp gene and chloramphenicol resistant was successfully introduced into two biocontrol agents Brevibacillus brevis ZJY-1 and Bacillus subtilis ZJY-116. After seed inoculation, the survival and colonization of the two strains were studied by periodically retrieving the GFP-tagged strains in the cucumber rhizosphere based on the selective markers. The results showed that both the strains could successfully colonize in the rhizosphere during the whole life of cucumber, and a higher colonization level was observed during anthesis and fruition stages. In pot trials, they could migrate to the nearby non-inoculated spontaneous weed plants, and reestablish in the rhizosphere of plants subsequently grown in the same pot. PMID- 16471357 TI - [Accumulation of phenolic acids in continuously cropped cucumber soil and their effects on soil microbes and enzyme activities]. AB - The study showed that in solar greenhouse continuously cropped cucumber soil, phenolic acids p-hydroxybenzoic acid, ferulic acid and benzoic acid had an obvious accumulation with increasing cropping year, and their contents were significantly higher after continuously cropped for 5 approximately 9 years than for 1 approximately 3 years. With the increasing concentration of treated exogenous phenolic acids, the amounts of bacteria, actinomycetes, total microbes, ammonibacteria, and nitrifying bacteria in cucumber root area increased first, but decreased then. Soil bacteria and actinomycetes had the largest amount at the concentration of 80 microg phenolic acids x g(-1) soil, while soil fungi (including Fusarium and Phytophthora) increased rapidly when the concentration of phenolic acids was lower than 120 microg x g(-1) soil. With increasing phenolic acids concentration, soil enzyme activities also increased first but decreased then, with the peak values differed in different treatments. PMID- 16471358 TI - [Screening of phytase-producing strain and its optimal solid state phytase producing conditions]. AB - Phytase can catalyze phytate hydrolyzed into inositol and phosphoric acid, and its application could make the phytic acid in animal feeds become more available, decrease feed cost, and at the same time, protect eco-environment. In this paper, a phytase-producing strain Trichoderma Viride LH374 was obtained through isolation and mutation, and its optimal conditions for solid state fermentation and phytase production were investigated. The results suggested that the optimal conditions of solid state fermentation were straw:rice bran ratio 8:2, medium initial pH 6.5, temperature 30 degrees C, culture duration 96 h, and moisture content 60%. Under the optimal conditions, LH374 could accumulate phytase up to 1,580 U x g(-1). PMID- 16471359 TI - [Heavy metals content and pH value in water environment of wild Chinese alligator (Alligator sinensis) habitat: a preliminary study]. AB - In order to reveal the reasons of the dramatic decline of wild Chinese alligator population, an investigation on its habitat status was made across the National Natural Conservation Regions of Chinese alligator in south Anhui Province from 2002 to 2003. Water samples from 21 sites were collected, and their heavy metals (Cu, Zn, Cd and Pb) content and pH value were measured. The results showed that the pH value was 7.84 +/- 0.62, with no significant difference between the waters with and without Chinese alligator, but with significant difference (P< 0.01) among 3 Chinese alligator habitats. In the waters with and without Chinese alligator, no significant difference was observed in the contents of test metals except Pb. The possible pollution source of Pb in wild Chinese alligator habitat was analyzed, which could provide reference for Chinese alligator conservation. PMID- 16471360 TI - [Effects of Cd contamination on paddy soil microbial biomass and enzyme activities and rice physiological indices]. AB - A pot experiment with rice under submerged condition showed that with the increase of Cd concentration, soil microbial biomass carbon (Cmic) and nitrogen (Nmic) increased initially but decreased at a certain concentration, and the turning points varied with different soil types. Soil enzyme activities had the similar variation trend with soil Cmic and Nmic, and the turning points varied with different soil types and soil enzymes. The variation coefficients were in order of dehydrogenase activity > acid phophatase activity > urease activity. Soil respiration rate and metabolic quotient increased tardily with increasing cadmium concentration. The chlorophyll content of rice increased initially but decreased then with the increase of Cd contamination, and the turning points differed with different soil types. Rice proline content and peroxidase activity were enhanced gradually with increasing cadmium concentration. The variation coefficients of rice physiological indices on paddy soils derived from silty loam and clayed red earth were in order of peroxidase activity > chlorophyll content > proline content, and peroxidase activity > proline content > chlorophyll content, respectively. Correlation analysis indicated that there was a close correlation between the variations of soil microbial biomass and enzymatic activities and rice physiological indices under Cd contamination. PMID- 16471361 TI - [Variation characteristics of leachate quality in a semi-aerobic municipal solid waste landfill]. AB - A large-scale semi-aerobic landfill set (21 m x 3.8 m x 6.0 m) was constructed to study the variation characteristics of pollutants in the landfill leachate. The results showed that after 39 weeks, the concentrations of CODCr, BOD5 and NH3+-N in the leachate declined rapidly, which reached 173, 30 and 15 mg x L(-1) respectively, and the reducing rate of NH3+-N was 99.6%. The pH value was below 7 during the first 2 weeks, but became weak alkaline after three weeks. Based on the experimental data, the attenuation equations of pollutants in the leachate were built. PMID- 16471362 TI - [Ecological toxicity of aluminum-based coagulant on representative corps in neutral environment]. AB - By the terrestrial ecotoxicological methods of seed germination and root elongation, and with aluminum-based coagulant AlCl3 which was widely used in wastewater- and water treatment as an example, this work studied the toxicological effects of aluminum-based coagulants on representative crops, including radish Raphanus sativus, cabbage Brassica chinensis and wheat Triticum aestivum, over a range of exposure concentrations under the neutral condition (pH= 7.0), with contrast to an acid condition (pH= 4.00). The results indicated that there was a significantly positive correlation between the concentration of AlCl3 and the inhibitory rate of root elongation and seed germination either under pH 7.0 or 4.0 (P < 0.01). Cabbage and wheat was more susceptive to AlCl3 toxicity under acid than under neutral condition, while radish was in adverse. The minimum inhibitory concentration of AlCl3 (< 2.0 mg x L(-1)) was lower under neutral than under acid condition. PMID- 16471363 TI - [Response of Vetiveria zizanioides to Pb2+ stress]. AB - The study on the eco-physiological indices of V. zizanioides under Pb2+ stress showed that with the increase of Pb2+ concentration (0 approximately 8 mmol x L( 1)) in water culture, the growth of V. zizanioides seedlings was badly influenced. Their leaves' conductivity increased, while SOD activity increased first and decreased then, but still higher than control. There was a positive correlation between SOD activity and Pb2+ concentration. POD and CAT activities were also increased first and decreased then, but when the Pb2+ concentration was > 4 mmol x L(-1), these enzyme activities were lower than the control, and had a significantly negative correlation with Pb2+ concentration. It was considered that the increase of enzyme activities was resulted from the adaptability of V. zizanioides stressed by Pb2+, while high concentration Pb2+ had an inhibitory effect on enzyme activities. PMID- 16471364 TI - [Impacts of Cd and Pb pollution on soil microbes in tobacco root zone and on tobacco leaf quality]. AB - A pot experiment was conducted to study the impacts of Cd and Pb pollution on soil microbes in tobacco root zone and the quality of tobacco leaves. The results indicated that the higher concentrations of Cd and Pb in soil, the less soil microbes in tobacco root zone. When the levels of Cd and Pb were < or = 10 and < or = 300 mg x kg(-1), respectively, the individuals of soil microbes decreased acutely. As for the quality indexes of tobacco leaves, the ratios of reductive sugar/nicotine and total-N/nicotine increased, and the main components became disharmony, which was disadvantage for the fragrance forming of tobacco leaves. The K content in tobacco leaves had a negative correlation with soil Cd and Pb concentrations, but a significantly positive correlation with the quantity of potassium bacteria in root zone under Cd pollution. No significant correlation was observed between the K content in tobacco leaves and the quantity of potassium bacteria in root zone under Pb pollution. PMID- 16471365 TI - [Environmental cost of rice production in Dongting Lake area of Hunan Province]. AB - To evaluate the environmental cost of rice production is of significance for the sustainable development of agriculture and the set-up of green GDP accounting system. In this paper, the environmental cost of rice production in Dongting Lake area, one of the main bread baskets in China, was evaluated from the aspects of pesticide pollution, chemical fertilizer pollution, greenhouse gases emission, plastic film persistence, paddyfield gleyization, and polderization. The results showed that in 1999, the total environmental cost of rice production in this area was 41.91 billion yuan, equivalent to 26.8% of its total agricultural production value, or 28.5% of the gross output of its planting industry. For the sustainable production of rice, the strategies for decrease environmental cost intrinsically were put forward. PMID- 16471366 TI - [Effects of illumination on overlying water quality and on phosphate adsorption by sediment]. AB - By the method of laboratory simulation, this paper studied the effects of illumination on the quality of overlying water and on the phosphate adsorption by sediment in a clean lake region. The results showed that under anaerobic condition, illumination could increase the pH value and the dissolved oxygen (DO) concentration of overlying water, while under aerobic condition, the concentration of dissolved inorganic phosphate (DIP) in overlying water declined fast at the beginning of the experiment, which indicated that DIP concentration was controlled by the DO concentration at the water-sediment interface. With the time went on, the DIP concentration in overlying water was much lower in the light than in the dark, indicating that illumination was the dominant controlling factor to the DIP concentration in overlying water. Phosphate could be adsorbed by the sediment with continuously loading, and the adsorbed amount varied with different environmental conditions. The effect of illumination on the phosphate adsorption by sediment was limited by the environment whether being aerobic or anaerobic. PMID- 16471367 TI - [N and P removal characters of eutrophic water body under planted float]. AB - The study on the N and P removal and N2O release of eutrophic water body under planted float Ipomoea aquatica and Oenanthe javanica showed that planted float had a good effect on the removal of N and P from eutrophic water body. The bioaccumulation of N and P by the plants accounted for 40.32% approximately 63.87% of the N and P removal, respectively, suggesting that plant uptake was the main removal process. Within the treating period, there was a rapid and sufficient nitrification but a slow and insufficient denitrification in the planted float system, and thereby, planted float system had a high NH4+-N removal rate and NO3(-)-N accumulation. The presence of planted float reduced the release flux of N2O, which was 17.14 microg N x m(-2) x h(-1) for Ipomoea aquatica, 37.38 microg N x m(-2) x h(-1) for Oenanthe javanica, and 85.08 microg N x m(-2) x h( 1) for the control. PMID- 16471368 TI - [Induction of pond-cultivated Anguilla japonica gonadial development by controlling temperature in high temperature season]. AB - In this study, the pond-cultivated Anguilla japonica was induced to mature by artificially controlling temperature at 18 +/- 2 degrees C in high temperature season (from June to September), with injection of hCG and CPE. The results showed that both the male and the female eel could be induced to mature, and the artificial maturation induction rate was 73.33% and 67.77%, while under non controlling temperature 25 approximately 32 degrees C, the maturation induction rate was 0 and 33.33%, respectively. In an ovulation induction test, the matured female eels were induced, with a spawning inducement rate of 70%, and a fertilization rate of 32%, which approached or reached the levels in low temperature season (with an average of 62.1% and 34.2%, respectively), and the zygote developed normally. At the temperature 22 and 24 degrees C, the zygote hatched to fry in about 39 h 15 min and 34 h 9 min. The experiment proved that it was possible to induce the eel to mature by controlling temperature in high temperature season, which provided some theoretical foundations for the techniques of artificially breeding eel in whole year. PMID- 16471370 TI - [Application value of organic ground surface mulch in urban forestry construction]. AB - This paper summarized the development process of organic ground surface mulch, and analyzed its important application value (such as the functions of improving soil physical and chemical properties, promoting plant growth, beautifying urban landscape, and preventing soil dust, etc.), and its values in protecting environment and benefiting economy. The problems in the application process of organic ground surface mulch were discussed, and the actual impendence and broad development space of its production and application in our urban forestry construction were pointed out. PMID- 16471369 TI - [Research advances in soil nitrogen cycling models and their simulation]. AB - Nitrogen is one of the necessary nutrients for plant, and also a primary element leading to environmental pollution. Many researches have been concerned about the contribution of agricultural activities to environmental pollution by nitrogenous compounds, and the focus is how to simulate soil nitrogen cycling processes correctly. In this paper, the primary soil nitrogen cycling processes were reviewed in brief, with 13 cycling models and 6 simulated cycling processes introduced, and the parameterization of models discussed. PMID- 16471371 TI - [Soil chemical property changes in vegetable greenhouse fields]. AB - To explore the changes of soil chemical properties in vegetable greenhouse, a comparative study was carried out with the samples gathered from vegetable greenhouse fields and their adjacent upland fields in Damintun Town, Xinming County, Liaoning Province. The results showed that compared with upland fields, the contents of soil organic carbon and total nitrogen in greenhouse fields increased significantly. At the depth of 0 approximately 30 cm, soil organic carbon in greenhouses of 1-, 4- and 10-year increased by 31.09%, 35.44%, and 66.80%, respectively, compared with the upland soil. Soil nitrate content at the depth of 0 approximately 30 cm in greenhouse fields was 5.05 approximately 12.49 times as much as that in upland fields. The nitrate content in different soil layers increased with the increasing age of greenhouse field., e.g., at the depth of 20 approximately 30 cm, soil nitrate content was significantly higher in 10 year than in 1- and 4-year greenhouse field, with an increase of 65.73% and 50.89%, respectively, and 6.55 times as much as that in upland field, which indicated that soil nitrate transported downwards, and obviously enriched in deeper soil layers under heavy application of fertilizer. Also with the increasing age of greenhouse field, soil pH decreased, while soil soluble salts accumulated. PMID- 16471372 TI - [Reconstruction of Polytrichum juniperinum population dynamics in a mire of China]. AB - By using 'history reconstruction method', this paper studied the dynamics of two Polytrichum juniperinum populations with and without sporophytes. The population with sporophyte production experienced a dynamic process from increase to stabilization, while that without sporophyte production experienced a process from increase to decrease. Before 2003, both the increase rate and the birth rate of two populations decreased continuously, with the death rate showing the tendency from decrease to a continuous increase. The advantages of 'history reconstruction method' in studying population dynamics were proved and discussed. PMID- 16471375 TI - [A treatment chart adapted to the arch form]. AB - Arch wires used in orthodontic treatment must conform to the shape of the dental arches so that in their final positions teeth will be set in a pathway of neuromuscular equilibrium. However, sometimes distortions of the alveoli deform the shape of the arch. In such cases the treatment charts constructed on the actual alveolo-dental relationship will not reflect desired objectives. We believe that the transverse muco-gingival dimension Andrews called the Wala line responds better to therapeutic requirements. Moreover, molar attachments, key elements of a full-banded appliance, are in harmony with it. So the type of chart that the authors propose for use with the Straight Wire technique, one based on the Wala line, will help orthodontists place teeth at the conclusion of orthodontic treatment in the best possible relationships to patients' specific anatomic configurations. PMID- 16471373 TI - [Evaluation of orthodontic friction using a tribometer with alternating movement]. AB - It is essential for orthodontists to control the complex phenomenon of friction. The in vitro techniques, usually dynamometers or tensile testing machines, used to measure the frictional resistance between arch wires and brackets are linear and unidirectional and can be criticised because tooth movements, such as tipping and uprighting, as well everyday oral activities, primarily chewing, are not uni dimensional but more closely resemble the small amplitude oscillatory phenomena known as fretting. We therefore decided to develop a fretting machine not with linear but with alternating movements better suited to evaluate the frictional behaviour of orthodontic bracket-wire combinations. Once we had completed construction of this device, we proceeded to measure the frictional resistance between one stainless steel bracket (MicroArch GAC) and five wires currently used in orthodontics (Two nickel-titanium shape memory alloys: Neo Sentalloy and Neo Sentalloy with Ionguard GAC--Three titanium-molybdenum alloys: TMA and Low Friction TMA Ormco and Resolve GAC). We were able to set up a classification of the wires according to their coefficient of friction, demonstrating the inefficacy of ion implantation and quantifying the increase in the coefficient of friction which occurs when Resolve wires are placed in the oral environment for approximately one year. PMID- 16471374 TI - [Distraction osteogenesis in orthodontic surgery of the mandible]. AB - Distraction osteogenesis is a bone creating procedure that can contribute materially to the treatment of Class II dento-skeletal disorders where the mandible is under-sized. Still, the serious commitments this treatment demands and the unpredictability of its occlusal results oblige practitioners to pinpoint precisely the indications for its use. We now believe that distraction osteogenesis can beneficially be employed for: 1--Adults with Class II malocclusions, a small mandible, and a lack of harmony between tooth and jaw size. This bone growing technique makes it possible to treat these patients without the extraction of premolars that would have been necessary in the traditional preparation for orthognathic surgery. 2--Patients whose mandibles are diminished in size in the area of the symphasis. Distraction osteogenesis of this anterior segment is the only way they can be treated without reduction of tooth structure or excessive labial tipping of teeth. 3--Some patients with short, or moderately short rami. We do not, at this time, believe that distraction osteogenesis is indicated for the treatment of any other kind of dento-facial deformity. But, in the future, improvement of surgical techniques and the miniaturization of appliances will doubtless extend its scope. PMID- 16471376 TI - [Orthodontic treatment need: the IOTN, or Index of Orthodontic Treatment Need (part 2)]. AB - IOTN is an index of orthodontic treatment need that has been devised on the basis of the available evidences of the deleterious effects of malocclusion on the aesthetic and health of the dentition. It has two components i.e. an aesthetic component and a dental health component. This index is widely used in epidemiological research and by third party payment for determination of those patients who should benefit from public funding. PMID- 16471377 TI - [The impact of socioeconomic status in the estimation of non-adult dental age]. AB - The aim of this study is to observe the possible impact of socioeconomic level on stages of macroscopic dental development. The sample consists of 456 children from European origin, aged 3.5 to 16 years (242 girls and 214 boys). We apply a radiographic method of non-adult dental age estimation based on Bayes' theorem. Both socioeconomic level and geographic origin of grandparents are given in a questionnaire devised by the first author. First, results obtained with the Bayesian dependent method (BD method) are compared to those obtained with a more commonly used method of dental age estimation based on Correspondence Analysis and linear Regression (CAR method). Second, two approaches are proposed in order to test potential relationships between socioeconomic level and stages of macroscopic dental development. Both are based on observed shifts between dental age and chronological age with regard to socioeconomic level. Results obtained with the BD method are better, in terms of quality, than those obtained with the CAR method. We observed no influence of socioeconomic level on stages of macroscopic dental development. PMID- 16471379 TI - [The Procuste superimpositions: its value in the study of typology and its development during Edgewise Tweed-Merrifield treatment]. AB - By transforming all figures into the same size, the Procruste mathematical superimpositions system allows orthodontists to analyze and compare geometric shapes whose different areas and forms would otherwise have made comparison impossible. Procruste is, accordingly, particularly useful in the study of facial types. In this retrospective study we analyzed the effects exerted by both growth and Edgewise Tweed Merrifield treatment upon changes in patients' typology of three different facial types. One hundred patients with Class II dental malocclusions were divided into three groups of deep bite, normal bite, and open bite. We found no significant statistical difference in the effects of treatment and of growth on modification of "form" of the facial skeleton as seen on profile cephalograms of the three categories. Patients in all the groups showed an average anterior rotation of the mandible. PMID- 16471378 TI - [Impacted canine tooth and the eyebrow: the orbit-eyelid impact of occlusal dysfunction]. AB - The orbital region and the dental occlusion are biomechanicaly interdependant. The maxillary basal bone is located in the orbital region due to the presence of the maxillary sinus. Any trouble of the dental occlusion such as a retained canine can involve a lack of projection of the supra and the infra-orbital rim. This lack of projection of the infra-orbital rim is responsible of lower eyelid fat pad; the lack of projection of the supra-orbital rim is responsible of an eyebrow-forehead ptosis. Clinical variations can be observed according to the side of the preeminent eye. PMID- 16471380 TI - Hurricanes, bird flu, and health policy. PMID- 16471381 TI - Office parties, drinking and driving, and employer liability. AB - Office parties are fun. They boost morale. They can also go over the line and result in company and personal liability. Holiday parties have ended up in court. This article explores "the line" between a good time and liability. What should you be aware of to protect your company and you? PMID- 16471382 TI - The new Medicare prescription drug benefit: important issues for physician practices. AB - Beginning January 1, 2006, Medicare beneficiaries will have the option of enrolling in a new Medicare program covering prescription drugs. This article summarizes the new benefit, suggests the factors beneficiaries and their physicians might consider when selecting a prescription drug plan, and outlines practice implications for physician offices. PMID- 16471383 TI - Embezzlement--don't let it happen to you! AB - Poor financial record keeping and blind trust are invitations for embezzlement. Clever thieves lure you into a false sense of security and complacency. They gain your trust, test the waters, and then go in for the kill. Don't be the victim of an embezzlement scheme that has your practice financing someone else's vacation. Embezzlement is often the result of a casual attitude toward handling finances, general neglect, or perhaps too much trust. PMID- 16471384 TI - Myriad pieces of facility development, Part 2. AB - The first article in this series discussed the complexity and breadth of deciding what to do when relocating your practice. Assuming you have accomplished those tasks, you now need to undertake the job of actually getting that done. This section takes you from the conceptualization of the project to the actual heavy lifting, and outlines the concrete (literally) steps that must be taken to bring the undertaking to fruition. PMID- 16471385 TI - The MPM pros answer the ten most commonly asked questions about: consultants. AB - The old joke is that a consultant comes to your office, borrows your watch, tells you what time it is, and then mails you a bill. The joke is funny because it is sometimes true; however, the right consultant can help you solve the right problems. Two experienced consultants tell you how to define the scope of the consultant's work, make an assessment of your internal capabilities, and determine the most appropriate way to structure payment. PMID- 16471386 TI - SEC and labor issue guidance on hiring plan advisers. PMID- 16471387 TI - Personal health records. AB - Nationwide, momentum is growing to provide patients with computer tools called personal health records (PHRs). These allow patients to participate in their own healthcare management by viewing, editing, or discussing their own medical data. Historically, PHRs targeted consumers, but contemporary PHRs are increasingly aimed at providers and payers. This article reviews the types of PHRs that are currently available, discusses the PHR functionalities that offer the best value for a medical practice, and provides strategies for making purchasing decisions. PMID- 16471388 TI - How to answer the most common zinger questions. AB - Patients are very likely to hit you with zingers--extremely tough questions that require all of your diplomacy and tact. How well you respond to the inevitable zingers depends in large part upon your preparation. This article describes what you must do to prepare yourself for your patients' zingers. It offers simple guidelines for responding to zinger questions, as well as how-to instruction that will enable you to hone your critical listening skills. This article also offers an effective strategy for preparing a list of potential zinger questions customized to your medical practice. It suggests ways to use zinger preparation as a staff training tool, relying in large part upon brainstorming, group work, and role-playing scenarios. Finally, this article presents the wrong way to answer zingers. Then, through sample dialogues, it offers much better and more effective responses that you might give your patients when you need to answer eight of the most common and difficult medical practice zingers. PMID- 16471389 TI - Minnesota agreement provides shape of things to come. PMID- 16471390 TI - Implementation of a touch-screen new patient registration system: a case study. AB - Medical practices are constantly seeking methods to increase their efficiency by using computer technology. The objective of this project was to assess to implementation of a touch-screen patient-entered data system for completion of intake paperwork required for new patients in a community-based behavioral healthcare clinic. The authors found that patients and staff across levels of computer experience, at the intervention clinic found the system easy to use and were highly satisfied with the experience. PMID- 16471391 TI - Reforming America's health system through innovation and entrepreneurship. AB - America's attempts for healthcare reform are gridlocked. Healthcare special interests are reluctant to abandon profitable activities, and American culture distrust of centralized federal power, belief in self-improvement, desire for choice, and belief in equal access to medical technologies-is slow to change. Physician entrepreneurship and innovation, coupled with consumer-driven healthcare and public-private partnerships, may break the present gridlock. PMID- 16471392 TI - Practices and hospitals, their physicians, and their noncompetes: breaking up should be hard to do. AB - The availability of specialty healthcare has become an issue of increasing importance. Practices and hospitals must be encouraged to assume the risk and responsibilities of expanding their fields of care. For their part, courts must give heavier weight to that aspect of the public interest in determining how their decision impacts access to specialty healthcare. In short, courts deciding the enforceability of noncompetition agreements must be willing to do something more than count doctors. PMID- 16471393 TI - Physician ancillary services: fad or fashion? Part 1. AB - Ancillary services are quite fashionable in today's medical practice, but perhaps you've wondered if they are only a fad. If you're considering adding services to increase your revenue, you're not too late; this trend seems here to stay. Doing your homework and due diligence is key to the success of your ancillary service. Before introducing a service into your practice or investing in an ancillary venture, make sure that you get off on sound footing. Major investments call for experienced consulting and tapping into financial, operational, and legal specialists to address specific needs. Part 1 of this two-part article defines ancillary services and addresses the following topics: performing the feasibility study; writing the business plan; and addressing the legal and regulatory issues. Part 2 will discuss financing arrangements, opening, operational, and ongoing management elements critical to success. PMID- 16471394 TI - J-1 physicians: a recruitment opportunity comes knocking. PMID- 16471396 TI - Let's not forget our children. PMID- 16471395 TI - Medical necessity and chart reviews. PMID- 16471397 TI - How to "un-muddy" the waters in the dental hygiene department. PMID- 16471398 TI - AlloDerm: an effective alternative to palatal donor tissue for treatment of gingival recession. PMID- 16471399 TI - An integrated systems approach to the placement of posterior composite resins. PMID- 16471400 TI - Adhesive cementation of chairside CAD/CAM inlays and onlays. AB - Chairside CAD/CAM restorations offer aesthetic, functional, biocompatible, and long-term successful alternatives to traditional materials and techniques, and can be fabricated in one appointment. Adhesive cementation is key for the long term clinical success of CAD/CAM inlays and onlays. The clinical use of a newly developed composite resin cement for adhesive cementation of CAD/CAM inlays/onlays has been described. PMID- 16471401 TI - Conversion from bar-retained to attachment-retained implant overdenture: case report. PMID- 16471402 TI - Sleuthing out hard-to-find canals. PMID- 16471403 TI - The direct resin onlay technique. PMID- 16471404 TI - Simplified restorative correction of the dentition using contact lens-thin porcelain veneers: a report of three cases. PMID- 16471405 TI - A technique for repair of a fractured porcelain-fused-to-metal bridge. PMID- 16471406 TI - Bite treatment, part 1: applying the "puppy dog sales technique" to dentistry. PMID- 16471407 TI - Charts, radiographs, study models: a forensic trilogy. PMID- 16471408 TI - Dental marketing 101. How and why to market your dental practice. PMID- 16471409 TI - Dentistry is a game: manage patient fear and win. PMID- 16471410 TI - Three key high-tech tools provide links to success. PMID- 16471411 TI - [Stroke of fate or personal challenge--subjective theories of illness as risk or protective factors in coping with chronic pediatric illness]. AB - Subjective illness theories have been neglected in the study of children's coping with chronic illness. It is hypothesized that a family's lay theory provides the cognitive framework of the coping process and thereby influences coping outcome. In a multimethod study, 79 children with insulin-dependent diabetes mellitus (age 6-12 years) and their parents were interviewed about their subjective illness theories, their coping behaviour and various outcome indicators. Several questionnaires measuring coping, family environment and health-related quality of life were also applied. Six types of subjective illness theories were identified. Families regarding diabetes as a daily strain or as a blow of fate showed particularly high levels of stress and lower quality of life compared to other groups. Families with inconsistent subjective illness theories were also at risk for lower quality of life. Ideas for prevention and intervention are presented at the end of the article. PMID- 16471412 TI - [Quality of life and chronic conditions: the perspective of children and adolescents in rehabilitation]. AB - Health related quality of life (QoL) is a relevant criterion to understand well being and function in children with chronic health conditions. Approaches to assessing quality of life are presented, based on a revision of the German KINDL questionnaire in the course of rehabilitation in three groups of chronically ill children (with diagnosis of asthma, atopic dermatitis or obesity), aged 8 to 17 years. Psychosocial and sociodemographic characteristics prior to, at the end and 12 months after rehabilitation were also assessed. At baseline, gender-, age- and diagnosis-specific impairments in QoL were found. Over time children improved, most clearly in children with obesity. Psychosocial predictors of QoL one year after rehabilitation were identified. The study shows that QoL assessment in children with chronic conditions is feasible and that children profit from rehabilitation programs. To further assess the QoL benefit in paediatric rehabilitation more studies are needed. PMID- 16471413 TI - [Family sense of coherence and resilience. A study on families with children with mental and physical disabilities]. AB - Drawing from concepts from family stress theory, resilience theory, Rolland's model of family adaptation to disability, and Antonovsky's salutogenetic model, this study explores the relationship between family sense of coherence and measures of family functioning, levels of stress, severity of the handicaps levels of functional impairment and demographic indicators. Participants were 37 families with children under 18 years with mental retardation and physical neurological disorders that consulted the Department of Pediatric Neurology. With a discriminant analysis, measures contributing to the separation into subgroups of families with high, medium and low family cohesion were identified. The measures permitted a satisfactory discrimination into groups in this clinical sample. The highest discriminatory power held sociodemographic variables, severity of impairment, as well as levels of family functioning and measures of family coping, which are potentially amenable to therapeutic change. The results indicate that family coherence can be regarded as a global resource contributing to family resilience, and therapeutic implications are being discussed. PMID- 16471414 TI - [How children experience a parent being chronically ill: a qualitative analysis of interviews with children of hemodialysis patients]. AB - Children's subjective experience of growing up with a somatically ill parent was studied by a qualitative analysis of individual interviews with children having a parent on hemodialysis. Transcripts of semi-structured interviews with 8 children and adolescents between 6 and 17 years and with one ill mother, whose father had had the same disease, were evaluated through structuring content analysis and psychoanalytical text interpretation. All individuals showed a strong sense of responsibility for their parents. In some cases this conflicted with their strives for autonomy. Almost all children tried to avoid conflicts with their parents. Younger children suffered from diffuse sadness, whereas children up from 11 years had concrete fears of medical complications or their parents' possible decease. In case parents spoke frankly about their illness, children felt relieved. Results suggest that growing-up with a chronically ill parent means a sustaining experience for a child, which requires enormous efforts of psychological adaptation. In preventive counselling concepts, parents should be strengthened in their competence as emotionally available attachment figures. PMID- 16471415 TI - [Chronically ill and psychologically impaired? Results of a new literature analysis]. AB - Based on earlier literature analyses, this study summarizes more recent research on the links between chronic illness and psychopathology. This analysis was conducted on specific types of illnesses such as diabetes, cancer, asthma and arthritis. Results show increased psychopathology among chronically ill children and adolescents in most studies. Duration of illness, severity of illness as well as gender unsystematically impacted this process. Noteworthy, too, is that some studies reported quite low psychopathology and even higher life quality in chronically ill children and adolescents. In some cases this can be considered as a sign of defense mechanisms. PMID- 16471416 TI - [Periradicular surgery with endoscopy--a clinical prospective study]. AB - Using an endoscope in periradicular surgery provides an impressive optical enlargement and may lead to better clinical diagnostics and a high rate of success. Aims of this investigation were examining this hypothesis as well as detecting influencing factors. In 2002 and 2003, 114 teeth of 91 patients were root-end resected using endoscope. Before surgery, the tooth, number of resected root-ends, content of root channels, clinical diagnosis and moment of surgery were recorded. Patients were controlled one year after surgery and success was evaluated as a summary of clinical and radiological findings. Four patients did not reappear for the control examination. Out of 110 teeth, treatment was successful in 91.8% and failed in 7.3%. 0.9% could not be classified. Statistically, more failures were noted after periradicular surgery in molars. The rate of success increased during the study by means of a learning curve. The results confirm the hypothesis that where using an endoscope periradicular surgery is highly successful. PMID- 16471417 TI - [Retention of fiber posts dependent on different resin cements]. AB - Tooth-coloured adhesive inserted fiber posts are used to restore endodontically treated teeth. In this investigation, the tensile bond strength of two different fiber posts systems (ER DentinPost and DT Light Post) in combination with five different resin cements was tested. The posts were inserted into artificial root canals in bovine dentin using Clearfil Core, RelyX Unicem, Panavia 21ex, Panavia F2.0 und Calibra. Titan posts (ER-Kopfstift), inserted with zincphosphate cement served as control group. ER DentinPost inserted with Clearfil Core had significantly higher tensile bond strengths than in combination with Panavia F2.0 (221.70 +/- 17.99 N) or Calibra (212.37 +/- 45.20 N). DT Light Post in combination with Calibra (338.20 +/- 46.40 N), Panavia F2.0 (321.69 +/- 40.11 N) and Panavia 21ex (290.41 +/- 55.28 N) showed significantly higher tensile bond strengths compared to adhesive cementation with RelyX Unicem (211.57 +/- 32.35 N) and Clearfil Core (131.67 +/- 51.72 N). The tensile bond strength of the control group was in the upper third of the values (315.43 +/- 51.38 N). Optical analysis of the post surface after decementation demonstrated in all cases adhesive cohesive mixed fractures. The adhesion of resin cement to the fiber posts and resin cement to root canal dentin is influenced by different factors. The combination of fiber post systems with the type of resin cement has a great influence on the tensile bond strength. PMID- 16471418 TI - [Fracture resistance of Procera Allceram depending on the framework design--an in vitro study]. AB - Procera AllCeram is one of the all-ceramic systems with an aluminium-oxide core employing CAD/CAM technology. The aim of the current study was to investigate the fracture resistance of Procera AllCeram full-ceramic crowns with a reduced core design compared to the conventional method. In addition, a possible influence of the preparation form (molars or premolars) and the cementation material (glas ionomer or composite) was analyzed. For both preparation forms, 30 ceramic cores with reduced margins (collarless cores, test) and 30 cores with extended cores (control) were veneered with porcelain in a standardized procedure (total 120 crowns). For the test group, Procera-AllCeram-margin ceramic material was used for the porcelain collar. 40 crowns each were cemented on stainless steel dies with either Ketac-Cem Aplicap or Panavia F. The additional 40 crowns were set on polyurethane dies without cementation and occlusally loaded until fracture occurred. Among the molar crowns, no differences were observed in fracture resistance neither for the different core designs (test or control) nor for the cementation materials. For the premolar form, fusing of a porcelain margin was associated with a reduction in fracture resistance, while the use of composite cement was accompanied with an increase. The present in vitro results indicate that for Procera AllCeram crowns with a highly undulating preparation margin, a conventional core design combined with adhesive cementation is preferable, especially in the posterior region due to higher chewing forces; this assumption needs to be proven in clinical studies. PMID- 16471419 TI - [Evaluation of the postoperative clinical results of repositioning isolated zygomatic fractures]. AB - This retrospective study included 148 patients who were evaluated for postoperative long-term results after osteosynthesis of isolated zygoma fractures. The influence of open reposition of the zygomaticoalveolar crest to middle face symmetry was evaluated quantitatively with a zygometer. In dependence on operative management patients were divided into two groups: In group I zygomaticofrontal suture (1-point-fixation) and infra-orbital rim were exposed (2 point-fixation), followed by osteosynthesis. In group II additional exposure of zygomaticoalveolar crest was performed and if required osteosynthesis was performed (3-point-fixation). For osteosynthetic reconstruction of isolated zygoma fractures the only 2-point-fixation is usually enough to achieve lateral middle face symmetry. In all cases zygomaticofrontal suture and alternatively infra-orbital rim or respectively zygomaticoalveolar crest should be supplied osteosynthetically. A further 3-point-fixation is indicated only in exceptional cases--for example comminuted fractures of lateral middle face columns. For an exact anatomical reposition of zygoma fractures the exposition and possible osteosynthesis of zygomaticoalveolar crest is recommended. PMID- 16471420 TI - [Halitophobia--an under-recognized clinical picture]. AB - A large part of the population suffers from halitosis. Causes can be both oral and non-oral changes. Coating of the tongue as well as marginal periodontitis are the most frequent oral causes. Non-oral causes are primarily tonsillitis and sinusitis but also general diseases as well as special diet habits. Non-real halitosis or halitophobia is unterstood by the compulsive idea to suffer from bad breath and to irritate others by this. This compulsive idea can change the complete pattern of behavior of these patients which in itself can lead to self isolation and even to suicide. Consultation hours for halitosis should be prepared for patients with non-real halitosis and build up corresponding interdisciplinary contacts. PMID- 16471421 TI - Nomenclature standardization for 'large pore size' virus-retentive filters. PMID- 16471422 TI - Modeling of drug release from celecoxib-PVP-meglumine amorphous systems. AB - An empirical assessment of drug release from amorphous systems of celecoxib (CEL), poly(vinyl pyrrolidone) (PVP), and meglumine (MEG) was performed and compared with that for its crystalline form. CEL-PVP (4:1 w/w) binary and CEL-PVP MEG (7:2:1 w/w) ternary amorphous systems provided higher drug dissolution. Mathematical modeling of drug release data was found to best fit the Hixson Crowell release model. The biphasic drug release during a 6-h duration exhibited higher release kinetics in the first phase due to the presence of drug in amorphous form. The release kinetics subdued in the latter phase due to ongoing devitrification process in amorphous systems. A comprehensive understanding of drug release from amorphous systems will accentuate the rationalized design of amorphous drug delivery systems. PMID- 16471423 TI - Bioadhesive tablets for controlled transdermal delivery of drugs. AB - Transdermal bioadhesive tablets were formulated by the direct compression method and evaluated. Carboxyfluorescein (CF) was used as a model permeant. The compression parameters were kept consistent and the composition of the formulation was varied to alter the release rate of the drug from the formulations. The dosage form was evaluated for physicochemical, adherence, and in vitro diffusion parameters. The transdermal flux of the drug decreased with the increasing ratio of Ethyl cellulose (EC)/hydroxypropyl cellulose (HPC). The test formulations were subjected to pharmacokinetic studies in mice. The formulations were able to maintain a steady state plasma concentration up to 12 hours. They were found to be safe for transdermal use as interpreted from skin irritation studies carried out on rabbits. PMID- 16471424 TI - Strategy for assessing the leachables impact of a material change made in a container/closure system. AB - Container/closure systems are extensively characterized in terms of their propensity to contribute leachable substances to the drug products they contain. Such a characterization is relevant until a change occurs in the composition or production of the container/closure system itself or the raw materials it is comprised of. When such a change occurs, it is necessary to ascertain the impact that the change would have on the validity and applicability of the previously performed leachables assessment. A general methodology for performing change control evaluations is developed in this manuscript and is illustrated via the use of a case study. PMID- 16471425 TI - Managing the microbiological quality of pharmaceutical excipients. AB - The microbiological quality of the pharmaceutical excipients used to manufacture pharmaceutical and over-the-counter drug products may significantly affect the outcome of individual processing steps and the microbiological attributes of the final drug products. Unlike active pharmaceutical ingredients, excipients are purchased from multiple suppliers and in many cases are produced for the food, cosmetics, consumer products, photographic, and paint industries and not specifically for the pharmaceutical industry, so the management of their microbiological quality is less straight- forward. This article discusses the qualification of suppliers, excipient production methods, compendial standards, regulatory controls, and microbial limits testing of excipients. Emphasis is given to risk assessment associated with pharmaceutical excipients. PMID- 16471426 TI - Determination of the electromechanical coupling factor of gallium orthophosphate (GaPO4) and its influence on resonance-frequency temperature dependencies. AB - The quartz homeotype gallium orthophosphate (GaPO4) is a representative of piezoelectric single crystals of large electromechanical coupling factor. It is known that its coupling factor kappa26 associated with the resonators vibrating in the thickness-shear mode is approximately two times greater than that of quartz. This property increases the spacing between the series and parallel resonance frequencies of resonators, as well as the difference between the resonance frequency temperature dependencies of the fundamental and harmonic resonance frequencies of resonators vibrating in the thickness-shear mode. In this paper, the methods for determination of the coupling factor kappa26 are presented, and the computed values are compared with the measured ones. The influence of the coupling factor to the resonance-frequency temperature dependencies of the fundamental and third harmonics of selected rotated Y-cut GaPO4 resonators vibrating in the thickness-shear mode is presented. The purely elastic case for a laterally unbounded plate, which corresponds closely to the limiting case of high harmonic resonance frequency-temperature behavior was assumed for the calculations. The computed temperature coefficients for the Y-cut orientation and calculated turnover point temperatures TTP for different (YX1) orientations are presented. PMID- 16471427 TI - Influence of test capacitor features on piezoelectric and dielectric measurement of ferroelectric films. AB - This paper presents both theoretical and numerical analyses of the piezoelectric and dielectric responses of a highly idealized film-on-substrate system, namely, a polarized ferroelectric film perfectly bonded to an elastic silicon substrate. It shows that both effective dielectric and piezoelectric properties of the films change with the size and configuration of the test capacitor. There exists a critical electrode size that is smaller than the diameter of the commonly used substrate. The effective film properties converge to their respective constrained values as capacitor size increases to the critical size. If capacitor size is smaller than the critical size, the surface displacement at the top electrode deviates from the net thickness change in response to an applied voltage because the film is deformable at the film/substrate interface. The constrained properties of the films depend only on those of bulk ferroelectrics but are independent of film thickness and substrate properties. The finding of the critical capacitor size together with analytical expressions of the constrained properties makes it possible to realize consistent measurement of piezoelectric and dielectric properties of films. A surface scanning technique is recommended to measure the profile of piezoresponses of the film so that the constrained properties of the film can be identified accurately. PMID- 16471428 TI - High power universal piezoelectric transformer. AB - This study describes a multilayer piezoelectric voltage and power transformer that has one direction poling, operates in a wide-frequency range and delivers both step-up and step-down voltages by inverting the electrical connections. In this design, the input and output electrodes are on the same side of the disk and are isolated from each other by a fixed isolation gap. The electrode pattern is a ring/dot structure such that it uses radial mode for both input and output part that are built-in on the same ceramic disk. A prototype transformer was fabricated of size 15 x 2.78 mm2 having mass of 3.8 gm. In the step-down configuration at the constant output power of 6 W, the transformer characteristics across a 100 omega load were found to be efficiency = 92%, gain = 0.21 input voltage = 110 V(rms), and temperature rise = 20 degrees C from the room temperature. In the step-up configuration at the constant output power of 5 W, the transformer characteristics across a 5 komega load were found to be efficiency = 97%, gain = 9.5, input voltage = 16 V(rms), and temperature rise = 8 degrees C from the room temperature. A detailed equivalent circuit analysis of the transformer was done, and the results were found to be in excellent agreement with the experimental results. PMID- 16471429 TI - The technical development of crystals and oscillators in China and their market situation. AB - In recent years the manufacture of and demand for crystals and oscillators in China have been increasing quickly because of the development of industries, communications, science, transportation, and electronic techniques. This paper introduces the manufacture, research, and characteristics of the technical development of crystals and oscillators in the last ten years in China. To meet the demand from industry, science, and other markets, the number of companies that produce and research crystals and oscillators has increased from several to more than 300 in the last 20 years. Manufacturing technology and equipment have also improved. For marketing and technical reasons, more than 75% of the companies manufacture mainly 49 micro, 49 micros, and microm crystals. Now more and more companies plan to build production lines to make cold-weld crystals and other precision crystals, and at least two companies have already built their production lines. Most of the precision crystals are AT-cut but some are SC-cut. Integrated TCXOs and DTCXOs have been made by different companies, and the frequency-temperature stability of them is from +/- 2 ppm to +/- 0.5 ppm in a wide temperature range. Specifications of some OCXOs are almost the same as those of foreign OCXOs. The main design techniques in China are experimental. However, in recent years, specially designed software has been developed for oscillator design, and advanced instrumentation and equipment are used. The market situation for crystals and oscillators has increased. PMID- 16471430 TI - Room temperature measurement of the anisotropic loss tangent of sapphire using the whispering gallery mode technique. AB - The anisotropic loss tangent has been determined in monocrystalline sapphire for components parallel and perpendicular to the crystal axis, using the whispering gallery (WG) mode method. The Q-factors of quasi-TE and quasi-TM modes were measured precisely in four cylindrical sapphire resonators at room temperature, from which was determined a maximum attainable Q-factor of (2.1 +/- 0.2) x 10(5) at 9 GHz in a quasi-TM mode. Sapphire dielectric material from three different manufacturers was compared over the 270-345 K temperature range and the 5-16 GHz frequency range. PMID- 16471431 TI - Effects of frequency-dependent attenuation and velocity dispersion on in vitro ultrasound velocity measurements in intact human femur specimens. AB - Numerous studies have shown that ultrasonic velocity measured in bone provides a good assessment of osteoporotic fracture risk. However, a lack of standardization of signal processing techniques used to compute the speed of sound (SOS) complicates the comparison between data obtained with different commercial devices. In this study, 38 intact femurs were tested using a through-transmission technique and SOS determined using different techniques. The resulting difference in measured SOS was determined as functions of the attenuation and the velocity dispersion. A numerical simulation was used to explain how attenuation and dispersion impact two different SOS measurements (group velocity, velocity based on the first zero crossing of the signal). A new method aimed at compensating for attenuation was devised and led to a significant reduction in the difference between SOS obtained with both signal processing techniques. A comparison between SOS and X-ray density measurements indicated that the best correlation was reached for SOS based on the first zero crossing apparently because it used a marker located in the early part of the signal and was less sensitive to multipath interference. The conclusion is that first zero crossing velocity may be preferred to group velocity for ultrasonic assessment at this potential fracture site. PMID- 16471432 TI - Improving accuracy in estimation of artery-wall displacement by referring to center frequency of RF echo. AB - Noninvasive measurement of mechanical properties, such as elasticity, of the arterial wall, is useful for diagnosis of atherosclerosis. The elasticity of the arterial wall can be estimated by combining measurement of displacement of the arterial wall with that of blood pressure. In general, the displacement of the arterial wall is estimated from the phase shift of radio frequency (RF) echoes between two consecutive frames using a correlation estimator with quadrature demodulated complex signals. Recently, digitized data of broadband RF echoes are available in modern diagnostic equipment. The Fourier transform can be used to estimate the phase of the RF echo at each frequency within the RF frequency bandwidth. Therefore, the phase shifts between RF echoes of two consecutive frames can be estimated at multiple frequencies. In this estimation, due to object displacement, the RF echo is time shifted in comparison with that of the previous frame. However, the position of the time window for the Fourier transform is not changed between two consecutive frames. This change in relative position between the RF echo and the time window has a strong influence on the estimation of the artery-wall displacement, resulting in error. To suppress this error, the phase shift should be estimated at the actual RF center frequency. In this paper, this error suppression was investigated through simulation experiments and in vivo experiments on the human carotid artery. PMID- 16471433 TI - Despeckling of medical ultrasound images. AB - Speckle noise is an inherent property of medical ultrasound imaging, and it generally tends to reduce the image resolution and contrast, thereby reducing the diagnostic value of this imaging modality. As a result, speckle noise reduction is an important prerequisite, whenever ultrasound imaging is used for tissue characterization. Among the many methods that have been proposed to perform this task, there exists a class of approaches that use a multiplicative model of speckled image formation and take advantage of the logarithmical transformation in order to convert multiplicative speckle noise into additive noise. The common assumption made in a dominant number of such studies is that the samples of the additive noise are mutually uncorrelated and obey a Gaussian distribution. The present study shows conceptually and experimentally that this assumption is oversimplified and unnatural. Moreover, it may lead to inadequate performance of the speckle reduction methods. The study introduces a simple preprocessing procedure, which modifies the acquired radio-frequency images (without affecting the anatomical information they contain), so that the noise in the log transformation domain becomes very close in its behavior to a white Gaussian noise. As a result, the preprocessing allows filtering methods based on assuming the noise to be white and Gaussian, to perform in nearly optimal conditions. The study evaluates performances of three different, nonlinear filters--wavelet denoising, total variation filtering, and anisotropic diffusion--and demonstrates that, in all these cases, the proposed preprocessing significantly improves the quality of resultant images. Our numerical tests include a series of computer simulated and in vivo experiments. PMID- 16471434 TI - Reduction of variance in spectral estimates for correction of ultrasonic aberration. AB - A variance reduction factor is defined to describe the rate of convergence and accuracy of spectra estimated from overlapping ultrasonic scattering volumes when the scattering is from a spatially uncorrelated medium. Assuming that the individual volumes are localized by a spherically symmetric Gaussian window and that centers of the volumes are located on orbits of an icosahedral rotation group, the factor is minimized by adjusting the weight and radius of each orbit. Conditions necessary for the application of the variance reduction method, particularly for statistical estimation of aberration, are examined. The smallest possible value of the factor is found by allowing an unlimited number of centers constrained only to be within a ball rather than on icosahedral orbits. Computations using orbits formed by icosahedral vertices, face centers, and edge midpoints with a constraint radius limited to a small multiple of the Gaussian width show that a significant reduction of variance can be achieved from a small number of centers in the confined volume and that this reduction is nearly the maximum obtainable from an unlimited number of centers in the same volume. PMID- 16471435 TI - Optimizing multicompression approaches to elasticity imaging. AB - Breast lesion visibility in static strain imaging ultimately is noise limited. When correlation and related techniques are applied to estimate local displacements between two echo frames recorded before and after a small deformation, target contrast increases linearly with the amount of deformation applied. However, above some deformation threshold, decorrelation noise increases more than contrast such that lesion visibility is severely reduced. Multicompression methods avoid this problem by accumulating displacements from many small deformations to provide the same net increase in lesion contrast as one large deformation but with minimal decorrelation noise. Unfortunately, multicompression approaches accumulate echo noise (electronic and sampling) with each deformation step as contrast builds so that lesion visibility can be reduced again if the applied deformation increment is too small. This paper uses signal models and analysis techniques to develop multicompression strategies that minimize strain image noise. The analysis predicts that displacement variance is minimal in elastically homogeneous media when the applied strain increment is 0.0035. Predictions are verified experimentally with gelatin phantoms. For in vivo breast imaging, a strain increment as low as 0.0015 is recommended for minimum noise because of the greater elastic heterogeneity of breast tissue. PMID- 16471436 TI - Real-time, 3-D ultrasound with multiple transducer arrays. AB - Modifications were made to a commercial real-time, three-dimensional (3-D) ultrasound system for near simultaneous 3-D scanning with two matrix array transducers. As a first illustration, a transducer cable assembly was modified to incorporate two independent, 3-D intra-cardiac echo catheters, a 7 Fr (2.3 mm O.D.) side scanning catheter and a 14 Fr (4.7 mm O.D) forward viewing catheter with accessory port, each catheter using 85 channels operating at 5 MHz. For applications in treatment of atrial fibrillation, the goal is to place the sideviewing catheter within the coronary sinus to view the whole left atrium, including a pulmonary vein. Meanwhile, the forward-viewing catheter inserted within the left atrium is directed toward the ostium of a pulmonary vein for therapy using the integrated accessory port. Using preloaded, phasing data, the scanner switches between catheters automatically, at the push of a button, with a delay of about 1 second, so that the clinician can view the therapy catheter with the coronary sinus catheter and vice versa. Preliminary imaging studies in a tissue phantom and in vivo show that our system successfully guided the forward viewing catheter toward a target while being imaged with the sideviewing catheter. The forward-viewing catheter then was activated to monitor the target while we mimicked therapy delivery. In the future, the system will switch between 3-D probes on a line-by-line basis and display both volumes simultaneously. PMID- 16471437 TI - Arbitrary waveform coded excitation using bipolar square wave pulsers in medical ultrasound. AB - This paper presents a new coded excitation scheme that efficiently synthesizes codes for arbitrary waveforms using a bipolar square wave pulser. In a coded excitation system, pulse compression is performed to restore the axial resolution. In order to maintain low range sidelobes, the system needs to transmit signals that have smooth spectra. However, such a transmitter requires the generation of arbitrary waveforms and, therefore, is more expensive. In other words, a trade-off is necessary between the compression performance and the transmitter cost. Here we propose a method that preserves the low-cost advantage of a bipolar pulser while achieving approximately the same compression performance as an arbitrary waveform generator. The key idea of the proposed method is the conversion of a nonbinary code (i.e., requiring an arbitrary waveform generator) with good compression performance into a binary code (i.e., requiring only a bipolar pulser) by code translation and code tuning. The code translation is implemented by sending the nonbinary code into a virtual one-bit, sigma-delta modulator, and the code tuning involves minimizing the root-mean square error between the resultant binary code and the original nonbinary code by sequential and iterative tuning while taking the transducer response into account. Tukey-windowed chirps are known to have good compression performance. Such chirps of different durations (16, 20, and 24 micros), all with a taper ratio of 0.15, a center frequency of 2.5 MHz, and an equivalent bandwidth of 1.5 MHz, were converted into binary Tukey-windowed chirps that were compared with pseudochirps (i.e., direct binary approximations of the original chirp) over the same spectral band. The bit rate was 40 MHz. Simulation results show that the use of binary Tukey-windowed chirps can reduce the code duration by 20.6% or the peak sidelobe level by 6 dB compared to the commonly used pseudochirps. Experimental results obtained under the same settings were in agreement with the simulations. Our results demonstrate that arbitrary waveform coded excitation can be realized using bipolar square wave pulsers for applications in medical ultrasound. PMID- 16471438 TI - Optical and acoustic detection of laser-generated microbubbles in single cells. AB - Acoustically monitored laser-induced optical breakdown (LIOB) has potential as an important tool to diagnose and treat living cells. Laser-induced intracellular microbubbles are readily detectable using high-frequency ultrasound, and LIOB can be controlled to operate within two distinct regimes. In the nondestructive regime, a single, short-lived bubble can be generated within a cell, without affecting its immediate viability. In the destructive regime, the induced photodisruption quickly can kill a targeted cell. To generate and monitor this range of bioeffects in real time, we have developed a system integrating an ultrafast laser source with optical and acoustic microscopy. Experiments were performed on monolayers of Chinese hamster ovary (CHO) cells. A 793 nm, 100 fs laser pulsed at 3.8 kHz was tightly focused within each cell to produce the photodisruption, and a 50 MHz ultrasonic transducer monitored the resultant bubble via continuous pulse-echo recordings. Photodisruption was also observed using bright field microscopy, and cell viability was assessed following laser exposure with a trypan blue assay. By controlling laser pulse fluence and exposure duration, either nondestructive or destructive LIOB could be produced. The intracellular position of the laser focus was also varied to demonstrate that cell viability was affected by the specific location of material breakdown. PMID- 16471439 TI - Ultrasonic contrast agent shell rupture detected by inertial cavitation and rebound signals. AB - Determining the rupture pressure threshold of ultrasound contrast agent microbubbles has significant applications for contrast imaging, development of therapeutic agents, and evaluation of potential bioeffects. Using a passive cavitation detector, this work evaluates rupture based on acoustic emissions from single, encapsulated, gas-filled microbubbles. Sinusoidal ultrasound pulses were transmitted into weak solutions of Optison at different center frequencies (0.9, 2.8, and 4.6 MHz), pulse durations (three, five, and seven cycles of the center frequencies), and peak rarefactional pressures (0.07 to 5.39 MPa). Pulse repetition frequency was 10 Hz. Signals detected with a 13-MHz, center-frequency transducer revealed postexcitation acoustic emissions (between 1 and 5 micros after excitation) with broadband spectral content. The observed acoustic emissions were consistent with the acoustic signature that would be anticipated from inertial collapse followed by "rebounds" when a microbubble ruptures and thus generates daughter/free bubbles that grow and collapse. The peak rarefactional pressure threshold for detection of these emissions increased with frequency (e.g., 0.53, 0.87, and 0.99 MPa for 0.9, 2.8, and 4.6 MHz, respectively; five-cycle pulse duration) and decreased with pulse duration. The emissions identified in this work were separated from the excitation in time and spectral content, and provide a novel determination of microbubble shell rupture. PMID- 16471440 TI - Enhanced heat deposition using ultrasound contrast agent--modeling and experimental observations. AB - Ultrasound contrast agents (UCA), created originally for visualization and diagnostic purposes, recently have been suggested as efficient enhancers of ultrasonic power deposition in tissue. The ultrasonic energy absorption by the contrast agents, considered as problematic in diagnostic imaging, might have beneficial impact in therapeutic applications such as targeted hyperthermia-based or ablation treatments. Introduction of gas microbubbles into the tissue to be treated can improve the effectiveness of current treatments by limiting the temperature rise to the treated site and minimizing the damage to the surrounding healthy tissues. To this end, proper assessment of the governing parameters of energy absorption by ultrasonically induced stabilized bubbles is important for both diagnostic and therapeutic ultrasound applications. The current study was designed to predict theoretically and measure experimentally the dissipation and heating effects of encapsulated UCA in a well-controlled and calibrated environment. The ultrasonic effects of the microbubble concentration, transmitted intensity, and frequency on power dissipation and stability of the UCA have been studied. The maximal temperature elevation obtained during 300 s experiments was 21 degrees C, in a 10 ml volume target containing UCA, insonifled by unfocused 3.2 MHz continuous wave (CW) at spatial average intensity of 1.1 W/cm2 (182 kPa). The results also suggest that higher frequencies are more efficiently absorbed by commonly used UCA. In particular, for spatial average intensity of 1.1 W/cm2 and concentration of 5 x 10(6) microspheres/cm3, no significant reduction of UCA absorption was noticed during the first 150 s for insonation at 3.2 MHz and the first 100 s for insonation at 1 MHz. In addition, when lower average intensity of 0.5 W/cm2 (160 kPa) at 3.2 MHz was used, the UCA absorptivity sustained for almost 200 s. Thus, when properly activated, UCA may be suitable for localized hyperthermic therapies. PMID- 16471441 TI - Three-dimensional phononic band gap calculations using the FDTD method and a PC cluster system. AB - This paper aims at studying the band gap phenomena of three-dimensional phononic crystals using the finite difference time domain (FDTD) method and a PC cluster system. In the paper, Bloch's theorem is applied to the wave equation and to the boundary conditions of the periodic structure. We calculate the variations of displacements and take discrete Fourier transform to acquire the resonances of the structures. Then, the dispersion relations of the bulk acoustic wave can be obtained and the band gaps are predicted accordingly. On the other hand, because of larger data calculation in three-dimensional phononic crystals, we introduce the PC cluster system and parallel FDTD programs written with respect to the architecture of a PC cluster system. Finally, we discuss the numerical calculation of two-dimensional and three-dimensional phononic crystals consisting of steel/epoxy and draw conclusions regarding the band gap phenomena between these phononic crystals. PMID- 16471442 TI - Dynamic measurement of amplitude-frequency effect of VHF resonators. AB - Dynamic amplitude-frequency effect measurements made of VHF quartz resonators indicate the presence of two separate mechanisms, the relative contribution of which varies for different cuts. For the SC-cut a step-change in drive level leads to a frequency shift with very short time constant (1 ms) that is presumably the classic direct nonlinear elastic effect. Others such as AT-, BT-, and LD-cuts show only a much longer time constant (200 ms), presumably a thermally related indirect nonlinear elastic effect. Near the SC-cut, resonators exhibit a combination of both short- and long-time-constant frequency changes. In some cases the two mechanisms produce frequency shifts in opposite directions. This behavior is problematic for some amplitude-frequency-effect measurement approaches because the measured shift depends on the timing of those measurements. It is not surprising that the SC-cut response shows only the direct nonlinear elastic effect, but it is unexpected that the AT- and BT-cuts appear not to show such effect. These results raise a question as to what amplitude frequency effect criterion is appropriate to use, as well as whether it is proper to determine nonlinear elastic constants from AT-cut resonator measurements that assume the effect is not thermally related. PMID- 16471443 TI - The study of an interaction of solid particles with various surfaces using TSM sensors. AB - The interaction of solid particles with various surfaces has been experiencing growing interest in many areas of nanotechnology, colloidal science, and biology. In this paper the interactions of solid particles with the surface of piezoelectric thickness shear mode (TSM) sensors have been studied. A mechanical model has been presented to evaluate the effect of particle loading on the behavior of a TSM sensor. The main sources contributing to the interaction, such as Van der Waals force, friction force, and electrostatic force, are discussed. Experiments have been designed for 10-100 microm particles on the 5-MHz and 10 MHz TSM sensors. It has been shown that the resonant frequencies of the TSM sensors might increase or decrease depending on the interaction conditions. The results have shown that the TSM sensor technique could provide the information on the mass/size of a particle and the binding energy between a particle and the sensor surface. This technique may find its applications in characterizing the properties of an interaction between particles and various surfaces. PMID- 16471444 TI - Comparative study of the nonlinear behavior of fatigued and intact samples of metallic alloys. AB - A method and an experimental setup to study the nonlinear behavior of fatigued and intact metal samples under high-frequency stresses has been developed and tested. The method is based on the measurement of the vibration velocity of stepped bar samples flexurally and/or extensionally vibrating at resonance. The vibration velocity signal, picked up by a laser vibrometer, is automatically acquired and analyzed by classical fast Fourier transform (FFT) methods for different excitation levels. The experimental setup was designed in such a way that it can be used for fatiguing and for testing the material samples. The tests have been done with samples of titanium and aluminium alloys. Results before and after fatigue crack nucleation are compared. A strongly nonlinear, nonclassical behavior is observed for fatigued samples. Theoretical interpretations of the different nonlinear behavior of standing waves in fatigued and intact samples are explored, and the results of the models are qualitatively compared with the experimental data. The models' approach provides an interpretation about the origin of nonlinearity in fatigued samples. PMID- 16471445 TI - Ultrasonic imaging of biofilms utilizing echoes from the biofilm/air interface. AB - Ultrasonic imaging of biofilms in water is difficult due to the very low contrast in acoustic impedance between the biofilm and water. In this paper, biofilms exposed to moist air are scanned through the substrate in order to obtain echoes from the biofilm/air interface. A 50 MHz scanning system was used to scan 1 mm x 1 mm areas of biofilms in a 10 microm grid pattern. Two fast Fourier transform (FFT) based methods for enhancement of the film thickness measurement resolution are compared. Using these techniques, the surface topography of biofilms with thickness less than the acoustic wavelength can be imaged. PMID- 16471446 TI - Minimizing the bulk-wave scattering loss in dual-mode SAW devices. AB - The losses arising from the scattering of SAW into bulk waves in the nonsynchronous areas of SAW devices are studied numerically using the boundary element method combined with the finite element method. As a reference structure, we use a typical one-port hiccup resonator on 42 degrees Y-LiTaO3. Strong scattering into bulk wave occurs in the central gap due to an abrupt change in periodicity. To reduce the scattering, we replace the gap with electrodes having reduced pitches. We show that it is possible to significantly increase the Q factor of the resonator while keeping the resonant frequency unchanged. Two types of structures are studied: the "distributed" gap and the "accordion" gap. To minimize the bulk-wave scattering in dual-mode SAW filters, we replace the metallized gaps in the traditional filter with distributed gaps. We find an optimal combination of pitch and metallization ratio in the gaps, reducing the insertion loss by 0.3 dB. PMID- 16471447 TI - Pure SH-SAW propagation, transduction and measurements on KNbO3. AB - Potassium niobate (KNbO3) supports the electromechanically active pure shear horizontal surface acoustic wave (SH-SAW) mode along Z-axis cylinder orientations, Euler angles (phi, 90 degrees, 0 degrees), in which two uncoupled wave solutions exist: a purely mechanical sagittal Rayleigh SAW and a piezoelectrically stiffened pure SH-SAW. Within this family of cuts, a maximum electromechanical coupling coefficient for the pure SH-SAW, K2 = 53%, is observed along (0 degrees, 90 degrees, 0 degrees). This pure SH-SAW orientation also has the maximum value of electromechanical coupling observed along rotated Y-cut X propagation directions, Euler angles (0 degrees, theta, 0 degrees). The use of the pure SH-SAW mode is attractive for liquid-sensing applications because the SH SAW is modestly attenuated by the adjacent liquid, unlike the generalized SAW (GSAW), which has particle displacement normal to the surface. This work investigates propagation and excitation properties of the SH-SAW and the shear horizontal bulk acoustic wave (SH-BAW) on single crystal KNbO3, Euler angles (0 degrees, 90 degrees, 0 degrees). Interdigital transducer (IDT) arrays are analyzed using boundary element method (BEM) techniques, addressing IDT properties such as: power partitioning between the SH-SAW and SH-BAW, SH-BAW radiation as a function of wave vector direction and radiation angle, and overall IDT impedance. The percentage of SH-SAW power to total input power is above 98% for IDTs containing 1.5 to 5.5 wavelengths of active electrodes with surrounding metalized regions. For nonmetalized regions outside the IDT, the ratio drops to between 1 and 2%, showing the importance of an energy trapping structure for efficient SH-SAW excitation and propagation along this orientation. Simulated and experimental IDT admittance results are compared, verifying the validity of the analysis performed. The reported measurements on the frequency variation with temperature indicate that the orientation considered is temperature compensated at about 8 degrees C. The surface of the SH-SAW devices fabricated have been loaded with deionized water and showed additional 1.6 dB transmission loss with respect to the unloaded surface, verifying the suitability of the pure SH-SAW mode on KNbO3 for liquid sensor applications. PMID- 16471448 TI - Theoretical analysis of micro-machined ultrasonic transducer using a simple 1-D model. AB - Micro-machined ultrasonic transducers (MUT) appear as an attractive alternative to standard bulk transducers mainly based on PZT ceramic actuators. However, the simulation of these new devices requires one to take correctly into account their operating conditions. Particularly, most of the MUT structures are periodic, associating a very large number of elementary actuators excited in phase. Furthermore, the development of an equivalent to the Mason model for MUTs would help in the promotion of this new kind of transducers. In this work, we propose a very simple model based on the material resistance theory to describe the flexural motion of a MUT. It is associated with a periodic Green's function development to take into account radiation in water. Basic working principles of MUT then are deduced from computing results, which coincides with already published data on that topic. PMID- 16471449 TI - Development of a 35-MHz piezo-composite ultrasound array for medical imaging. AB - This paper discusses the development of a 64-element 35-MHz composite ultrasonic array. This array was designed primarily for ocular imaging applications, and features 2-2 composite elements mechanically diced out of a fine-grain high density Navy Type VI ceramic. Array elements were spaced at a 50-micron pitch, interconnected via a custom flexible circuit and matched to the 50-ohm system electronics via a 75-ohm transmission line coaxial cable. Elevation focusing was achieved using a cylindrically shaped epoxy lens. One functional 64-element array was fabricated and tested. Bandwidths averaging 55%, 23-dB insertion loss, and crosstalk less than -24 dB were measured. An image of a tungsten wire target phantom was acquired using a synthetic aperture reconstruction algorithm. The results from this imaging test demonstrate resolution exceeding 50 microm axially and 100 microm laterally. PMID- 16471451 TI - Thickness-shear vibrations of rotated Y-cut quartz plates with imperfectly bonded surface mass layers. AB - A solution is obtained from the three-dimensional equations of linear piezoelectricity for the pure thickness-shear vibration of rotated Y-cut quartz or langasite plates with imperfectly bonded surface mass layers. The solution includes a few results in the literature as special cases. It is shown that the mass layers lower the resonant frequencies when they are relatively perfectly bonded to the crystal surface, and that loosely bonded mass layers may raise the frequencies. The results are useful in the analysis of frequency stability of quartz resonators and acoustic wave sensors. PMID- 16471450 TI - Modeling and design optimization of large-deflection piezoelectric folded cantilever microactuators. AB - This paper presents a novel, large-deflection piezoelectric folded cantilever microactuator (PFCM). A multimorph model for the large-deflection PFCM is derived, in which unified formulas for deflection angle and vertical displacement of N-level PFCM are obtained. Based on the model, multilayer PFCMs using PZT film are designed and optimized. The large-deflection PFCM is extensively applicable to micro-devices or microsystem applications such as micro-optical switches, microelectromechanical systems (MEMS) scanners, and so forth. PMID- 16471452 TI - The Gaussian beam expansion applied to Fresnel field integrals. AB - A general result of the Gaussian angular-spectrum expansion is presented for computing the Fresnel field integral. The reciprocity relation of the Gaussian expansion parameters in both coordinate and angular-spectrum domains is obtained. Some examples are provided that demonstrate the usefulness of this relation. PMID- 16471453 TI - Surface acoustic wave velocity in single-crystal AlN substrates. AB - The surface acoustic wave velocity has been measured on a-plane (c-propagation) and c-plane oriented bulk aluminum nitride (AlN) single crystals using the S11 parameter method in the frequency range 160-360 MHz. The SAW velocity is 5760 m/s for both orientations. From comparison of this value with the simulations using various elastic constants of AlN available in literature, we estimated the elastic constant C44 to be 122 +/- 1 GPa. PMID- 16471454 TI - Review of a treatment for psoriasis using herose, a botanical formula. AB - This study was undertaken to assess the efficacy of the herose formula as a treatment for psoriasis patients who have been previously treated with immunosuppressants. The study was an open one conducted in one center. Fifteen patients were instructed to take four herose capsules (450 mg per capsule) three times a day for 10 months. At each visit the investigator assessed each patient's PASI and therapeutic response to the treatment. The patients were treated and observed from February of 2004 to November of 2004. The results showed that the herose capsule may be an effective and safe treatment for moderate to severe plaque psoriasis. PMID- 16471455 TI - Basal cell carcinoma in Turkey. AB - Basal cell carcinoma (BCC) is the most common form of cancer in Caucasians. This study was undertaken to define the clinical features of BCC treated by surgical excision in Turkish patients. A group of 198 patients with 216 BCCs, all treated by surgical excision, were studied prospectively. The age and sex distribution, personal and family histories of the patients, skin type, sites and sizes of the lesions, clinical types of the lesions, status of the surgical margins, and incidences of recurrence were analyzed. The male to female ratio was 0.9, and the average age of the patients was 64.5 years. Of the tumors, 91.5% occurred on the head and neck. There were also personal and/or family histories of cancer in 24.8% of our cases. Of all BCCs treated, 83.8% had a noduloulcerative clinical appearance. The vast majority of our patients were Fitzpatrick type 3 (53.0%) and type 2 (41.5%). There were seven recurrences after follow-up periods varying between three months and two years. In conclusion, the clinical characteristics of BCC in Turkish patients have both similarities and differences compared to those of other countries. PMID- 16471456 TI - Contact allergies to cosmetics: testing with 52 cosmetic ingredients and personal products. AB - Fifty patients of both sexes with clinically suspected cosmetic dermatitis were subjected to patch testing with a cosmetic and fragrance series, approved by the Contact and Occupational Dermatosis Forum of India (CODFI), and with selected allergens from the Indian Standard Series (ISS). Most of these patients were young adults between 10-29 years; the mean age was 27.5 years. The majority of the patients had cosmetic dermatitis of <1 year duration (68%). The occupational profiles of the patients included students (46%), housewives (18%), teachers (10%) and laborers (4%). A miscellaneous group, comprised of tailors, farmers, staff nurses, beauticians, jewellers and engineers, accounted for the remaining 22%. The most commonly involved site was the face, followed by the forehead, neck and scalp. Patch testing of these patients revealed that, out of the 50 subjects tested, thirty-three (66%) reacted to one or more allergens. Fragrance components were the most common offending allergen (51.5%) followed by preservatives (39.3%), paraphenylenediamine (PPD) (21.2%), and cetrimide and tertiary butyl hydroquinone (12.1% each), in descending order of frequency. Hence, patch testing, with the standard series supplemented by personal cosmetics; should be considered for patients with cosmetic dermatitis to determine the offending allergen so as to avoid further contact with that allergen. PMID- 16471457 TI - A novel computer-assisted algorithmic method for differential diagnosis of dermatological diseases. AB - To deal with human surface locations electronically, it is essential to establish a designation method to code the sites. For the development of site designation methods and databases, dermatological textbooks, atlantes and clinical photographs were collected and reviewed. The sites were systematically named, numbered, and mapped on human models. The whole human surface was divided into 484 segments, and then assembled into 15 groups and 29 subgroups. After establishing the site designation method, we constructed a database of the sites of occurrence of dermatological diseases, and a database of age, sex, signs and symptoms. More than 700 diseases were analyzed and coded into the database. To gather information on the site of a particular disease, not only textbooks, and atlantes of dermatology, but also photographs obtained during clinical practices were reviewed, as clinical photos of patients can be more delicate than the descriptions in textbooks. The prototype of a computer assisted dermatological diagnosing system was made using site a designation method and the databases. This interactive computer program worked relatively well as a helpful guide or dermatological diagnosing tool. Further diseases are being added to the databases. PMID- 16471458 TI - Efficacy of triamcinolone acetonide in various acquired nail dystrophies. AB - The treatment of nail disorders is currently an unsatisfying exercise. Isolated nail involvement generally does not warrant any systemic therapy. At the same time, treatment is requested because of significant cosmetic and functional handicap. Intralesional triamcinolone acetonide (TA) in the proximal nail fold was evaluated as a treatment modality in 30 patients with twenty-nail dystrophy, 14 with nail lichen planus, and 6 with nail psoriasis. The number of involved nails varied from 1-20, and 1-10 nails were treated with TA. Fourteen patients discontinued treatment after 1-2 sittings. Out of the 28 patients completing the treatment protocol, 16 showed 75-100% improvement. Predominant side effects included pain, subungual hematoma formation, proximal nail fold hypopigmentation, and atrophy. TA given as a single injection in the proximal nail fold produced good improvement in a significant number of patients completing the treatment protocol. Lower concentrations of TA (5 mg/ml) were quite effective in treating various dermatoses affecting the nail unit. Our technique had fewer side effects than needle-less injection or multiple injection techniques. Careful attention to injection technique further minimized the side effects associated with the procedure. Sixteen patients completed the six-month follow-up and a relapse of nail changes was seen in 10. The relapses were equally responsive to retreatment. TA injected into the proximal nail fold area is a useful, cheap and efficacious treatment for dermatoses affecting the nail unit. PMID- 16471459 TI - Localized periorbital edema induced by Ibuprofen. AB - We documented localized periorbital edema in one patient with ibuprofen sensitivity without underlying chronic urticaria. The reaction developed one hour after ingestion of 200 mg of ibuprofen. No systemic symptoms were observed. No other NSAIDs did not induce symptoms. This patient was able to tolerate doses of ibuprofen after pretreatment with terfenadine. These observations suggest that histamine played a central role in this ibuprofen-induced skin reaction. Treatment with terfenadine enabled the patient to tolerate ibuprofen without experiencing any side effects. To the best of our knowledge, this is the first reported case of periorbital edema induced by ibuprofen. PMID- 16471460 TI - A case of lichenoid drug eruption associated with sildenafil citratus. AB - A 53-year-old man developed lichenoid lesions on the upper chest, posterior surfaces of the trunk, and abdominal region about three months before his first visit. Physical examination and laboratory findings were normal; histopathology showed vacuolar degeneration of basal keratinocytes in association with a dense lympho-histioid infiltrate arranged in a lichenoid pattern with a few melanophages and eosinophils. The fact that our patient had been irregularly taking sildenafil citratus (Viagra) led to the hypothesis of a lichenoid drug induced eruption. Our hypothesis was confirmed by clinical resolution three weeks after discontinuation of sildenafil citratus; moreover, the patient avoided the drug for about four months, and the eruption didn't reappear. Subsequently, we performed a challenge test with the drug, and the patient developed similar lichenoid lesions. Lichenoid eruptions are rather common dermatoses that can be induced by a great number of environmental agents and are clinically but not pathogenetically well defined. We report the present case because, despite the great number of drugs that can be implicated in the development of lichenoid eruptions, the association of such dermatoses and sildenafil citratus had been described only once previously in the literature until now. Furthermore, we wish to remark on the significance of a detailed anamnestic history to make the correct differential diagnosis between lichenoid drug-induced eruptions and lichen planus. This has a great clinical importance because simple discontinuation or substitution of the drug causes lichenoid drug-induced eruption resolution. PMID- 16471461 TI - Drug-induced hypersensitivity syndrome associated with human herpesvirus 6 and cytomegalovirus reactivation. AB - We describe a patient with drug-induced hypersensitivity syndrome (DIHS) associated with human herpesvirus 6 (HHV-6) and cytomegalovirus (CMV) infection induced by sulfasalazine. Two weeks after starting sulfasalazine to treat a rectal ulcer, the patient developed disseminated macular erythema accompanied by fever, liver injury, and lymphadenopathy. Seroconversion of antibodies to HHV-6 was observed. Systemic steroid treatment was not effective against the eruptions. Five months after the onset, he presented with an acute febrile disease. The detection of CMV antigen on peripheral blood leukocytes and positive staining for CMV on cutaneous endothelium indicated active CMV infection. Furthermore, he developed a bacteremia of methicillin resistant Staphylococcus aureus. An association the CMV reactivation with DIHS was suggested, although there remains the possibility that the systemic steroid treatment precipitated CMV reactivation. Recently, HHV-6 has been documented to have immunomodulating effects and to be associated with CMV reactivation. Therefore, we should pay attention to the possibility of CMV reactivation in patients with DIHS in whom the immunomodulating virus of HHV-6 has been reactivated. PMID- 16471462 TI - Primary anetoderma: a case report and its modified classification. AB - A fifty-nine-year-old healthy male presented with fifteen round pouches around his bilateral shoulders and proximal thighs, at which elasticity was lost on palpation. Histopathological examinations demonstrated that the lesional dermis was thinner than normal skin and there was loss of elastic fibers and mild inflammatory cell infiltration. Because there was no preceding inflammatory skin lesion or associated diseases, the patient was diagnosed with primary anetoderma. This is a rare case of primary anetoderma that showed loss of elastic fibers and the thinner dermis. In addition, a modified classification is proposed considering associated diseases with a review of literature. PMID- 16471463 TI - Plasma cell cheilitis treated with intralesional injection of corticosteroids. AB - Plasma cell cheilitis is an idiopathic, benign, inflammatory condition characterized by a dense band-like infiltrate of plasma cells in the oral mucosa. In addition to the lips, the disease can affect the penis, vulva, buccal mucosa, palate, gingiva, tongue, epiglottis, and larynx. Some authors have shown the effectiveness of topical or intralesional corticosteroids; however, there have been many reports describing poor therapeutic responses to topical steroids. We describe three patients with plasma cell cheilitis whose clinical condition responded rapidly to the intralesional injection of corticosteroids. PMID- 16471464 TI - A case of angiolymphoid hyperplasia with eosinophilia (ALHE) of the upper lip. AB - Angiolymphoid hyperplasia with eosinophilia (ALHE) is clinically characterized by intradermal or subcutaneous papules and/or nodules usually occurring in young adults. Lesions in the oral mucosa are extremely rare. We report a case and review the literature of ALHE cases involving the oral mucosa. A 40-year-old man presented with a painless, 20 x 20 mm, submucosal nodule on the upper lip. Histological examination of lip biopsy specimens revealed an increase in many small vessels. The vascular walls consisted of prominent endothelial cells with a histiocytoid appearance, which protruded into the lumen. Many eosinophils and lymphocytes were also seen around the vessels. The diagnosis of ALHE was made from the above findings. PMID- 16471465 TI - Disseminated lobular capillary hemangioma: two case reports. AB - Lobular capillary hemangioma, also known as pyogenic granuloma, is a common, solitary, benign neoplasm of the skin and mucous membranes. The etiology of this lesion remains unknown. Lobular capillary hemangioma can present rarely in a disseminated form, usually associated with other disorders. Two patients, aged 17 and 33, were admitted to the dermatology clinic with disseminated lobular capillary hemangiomas which appeared suddenly. There was no history of trauma in either case. The histopathology of the lesions was consistent with lobular capillary hemangioma. No associated disorders were found in physical and laboratory examinations other than mental retardation in one patient and a small hemangioma in the liver in the other. We report two cases of disseminated lobular capillary hemangioma without an associated disorder. PMID- 16471466 TI - A case of extensive linear porokeratosis with evaluation of topical tretinoin versus 5-flourouracil as treatment modalities. AB - A 19-year-old male presented with linear raised lesions over extensive areas of his body that followed Blaschko's lines. The lesions were asymptomatic with ridged borders. Histopathology confirmed a diagnosis of porokeratosis. He was given topical tretinoin and topical flourouracil for local application on specified areas. The cosmetic acceptability and the safety profile of tretinoin were found to be better than those of flourouracil. However, both the agents were equally efficacious. A rare case of extensive porokeratosis with comparative evaluation of two treatment modalities is hereby presented. PMID- 16471467 TI - Treatment of multiple lesions of Bowen's disease and squamous cell carcinoma with topical imiquimod. AB - We observed complete clinical and histopathologic remission of papules, plaques and nodules in two patients with multiple lesions of Bowen's disease and squamous cell carcinoma. Our findings indicate that topical imiquimod is a potentially promising therapeutic approach for the local treatment of early forms of cutaneous neoplasia, such as Bowen's disease and squamous cell carcinoma because it is a non-invasive therapy with a good cosmetic outcome. It may therefore be worthy of consideration in a subgroup of patients who cannot have or do not wish to have surgery. PMID- 16471468 TI - Squamous cell carcinoma complicating an untreated chronic discoid lupus erythematosus (CDLE) lesion in a black female. AB - A 36-years-old female presented with hypertrophic, depigmented plaques on both forearms and a fungating cauliflower-like growth on the left forearm. Histologically, the plaque lesions were discoid lupus erythematosus, and the fungating growth was a well-differentiated squamous cell carcinoma. PMID- 16471469 TI - Paraneoplastic pemphigus associated with pelvic inflammatory fibrosarcoma: a case report. AB - A 36-year-old African-American woman presented with an extensive stomatitis and pigmented cutaneous macules on the neck, axillae and hands. Subsequently she developed violaceus papules on the dorsa of the hands, histologically consistent with an interface dermatitis. After 18 months of progressive disease, paraneoplastic pemphigus was suspected and a search for an underlying neoplasm was initiated. An exploratory laparotomy revealed a pelvic mass and the histologic examination showed an inflammatory fibrosarcoma. The evidence of acantholysis on new cutaneous lesions and the positivity of indirect immunofluorescence with rodent urinary bladder epithelium reinforced the diagnostic criteria for paraneoplastic pemphigus, which is confirmed by the identification of strong protein bands at 210, 190 and 170 kd by immunoprecipitation. Paraneoplastic pemphigus should be considered when investigating atypical mucocutaneous manifestations of pemphigus vulgaris and lichen planus. Diagnostic screening for paraneoplastic pemphigus and a search for an underlying tumor should be performed. PMID- 16471470 TI - Bullous Systemic Lupus Erythematosus as an Initial Manifestation of SLE. AB - Bullous systemic lupus erythematosus (BSLE) is a rare subset of systemic lupus erythematosus that is often associated with autoimmunity to type VII collagen. We describe a 45-year-old woman with BSLE who presented with vesiculobullous lesions as an initial manifestation of SLE. The patient first noticed a widespread urticarial, erythematous eruption associated with tense blisters, erosions, and crusting. She was diagnosed with bullous pemphigoid and underwent a one-month course of treatment with betamethazone. Because of the appearance of marked proteinuria, a subsequent renal biopsy, and serological tests, the patient was diagnosed with rapidly progressive glomerulonephritis and systemic lupus erythematosus. The patient's IgG circulating antibodies labeled the dermal floor of salt-split skin and recognized type VII collagen in immunoblot studies. Although methylprednisolone pulse therapy for glomerulonephritis did not alleviate the vesicullobullous eruption, treatment with dapsone resulted in dramatic disappearance of the lesions. Cessation of dapsone therapy due to hemolysis with Heinz-body formation did not aggravate the bullous disease. Our case illustrates that a generalized vesiculobullous eruption can be the sole presenting manifestation of SLE. It also emphasizes the close temporal relationship between BSLE and lupus nephritis. PMID- 16471471 TI - A case of nodular scleroderma. AB - Nodular scleroderma is a rare complication of systemic sclerosis; the pathogenetic implications are still unknown, although many factors are supposed to play a role in lesion development. We report the case of a young woman suffering from systemic sclerosis, who developed nodular lesions during therapeutic management with D-penicillamine and plasmapheresis. In order to better understand the essence of this disease, we examined all the possible pathogenetic mechanisms that could be implicated in nodular lesion development. PMID- 16471472 TI - Initial cutaneous manifestations associated with histopathological leukocytoclastic vasculitis in two patients with antiphospholipid antibody syndrome. AB - Antiphospholipid antibody syndrome (APS) is a multisystem disorder associated with a variety of circulating autoantibodies that target different phospholipid protein complexes. APS is sometimes lethal as a result of severe sequelae, which may be primary or secondary to the underlying disease. We report two women who presented histopathologically with leukocytoclastic vasculitis as the first cutaneous manifestation and were subsequently diagnosed with APS associated with systemic lupus erythematosus (SLE). Patient 1 presented with widespread cutaneous necrosis (WCN) with rapidly spreading pain down the lower extremities. Skin biopsy specimens from her leg purpura and WCN revealed perivascular infiltrates with neutrophils consistent with leukocytoclastic vasculitis and thromboses of small-sized dermal vessels. Patient 2 exhibited livedo reticularis, painful cutaneous nodules with necrosis, ulcer, and erythematous macules on her lower extremities, shoulder, and face. Skin biopsies of her right knee showed intravascular thrombosis of small dermal vessels and infiltration of perivascular tissues with necrotizing granulomatous vasculitis in the dermis. We found that these various cutaneous manifestations with leukocytoclastic vasculitis were present at an early stage of APS. Although progression to leukocytoclastic vasculitis in patients with APS is uncommon, our data suggest that the association between microvascular occlusions and cutaneous vessel vasculitis has a predictive value for the pathogenesis. It is important for dermatologists to recognize these cutaneous signs to permit early and accurate diagnosis and treatment. PMID- 16471473 TI - Mannose-binding lectin (MBL) single nucleotide polymorphism is not associated with atopic dermatitis in Japanese patients. PMID- 16471474 TI - A solitary fibrous tumor arising in the perioral region. PMID- 16471475 TI - Polyglandular autoimmune syndrome associated with pigmented basal cell carcinoma. PMID- 16471476 TI - Generalized lichen sclerosus et atrophicus: report of a case. PMID- 16471477 TI - Cutaneous focal mucinosis: a case report. PMID- 16471478 TI - Dystrophic calcinosis cutis following chest wall and breast irradiation. PMID- 16471479 TI - The Effects of Food Treat Provisioning and Human Interaction on the Behavioral Wel1-being of Rhesus Monkeys (Macaca mulatta). PMID- 16471480 TI - The Lightweight. Modular and Autodavable HEPA Filter Assembly for Isolation Racks Used in Animal Studies with Infectious Agents Requiring Biosafety Level 2 or 3 Containment. PMID- 16471481 TI - Determination of animal facility staffing levels. PMID- 16471482 TI - Plans for the In-house Construction of Environmental Enrichment Devices. PMID- 16471483 TI - A Veterinary Technician's Role in a Major Orthopedic Investigation: Evaluation of an Anterior Cruciate Ligament Prosthesis. PMID- 16471485 TI - Synthesis of dumbbell-shaped manganese oxide nanocrystals. AB - Dumbbell-like homogeneous MnO nanocrystals are obtained for the first time via the pyrolysis of manganese formate in trioctylamine/oleic acid media and the orientation aggregation mechanism is proposed for this 1D growth route. PMID- 16471486 TI - Cross-correlated relaxation between H1' chemical shift anisotropy and H1'-H2' dipolar relaxation mechanisms in ribonucleosides: application to the characterization of their anomeric configuration. AB - Cross-correlated nuclear spin relaxation between 1H chemical shift anisotropy (CSA) and 1H-1H dipolar relaxation mechanisms in ribonucleosides in solution phase are observed and used to identify their anomeric configuration. Only alpha ribonucleosides showed the presence of cross-correlated spin relaxation through differential spin-lattice relaxation (T1) of the H1' doublet. Dependence of the magnitude and the orientation of the H1' CSA tensor values on the glycosidic torsion angle and the fast time-scale internal motions present in the ribose moiety play a significant role in the characterization of the anomeric configuration of the nucleosides via cross-correlated relaxation. PMID- 16471487 TI - Why is formate synthesis insensitive to copper surface structures? AB - Experiments have revealed that formate synthesis from carbon dioxide and hydrogen is structure insensitive to copper catalyst surfaces, while the reverse formate decomposition reaction is structure sensitive. The present ab initio density functional theory (DFT) calculations show that the reaction of CO2 with surface atomic hydrogen initially leads to the formation of unstable monodentate formate, which has similar adsorption energies on Cu(111), Cu(100), and Cu(110). The structure of the transition state is similar to that of monodentate formate. It is also shown that gaseous CO2 is directly reacted with surface hydrogen, as suggested by previous experiments. The position of the similar transition state and the direct reaction mechanism well explain the similar energetic pathways, that is, the structure insensitivity. PMID- 16471488 TI - Competing magnetic interactions in a dinuclear Ni(II) complex: antiferromagnetic O-H...O moiety and ferromagnetic N3- ligand. AB - Synthesis, structural characteristics, magnetic studies and DFT calculations in Ni(II) dinuclear complexes containing two bridging N3- and an O-H...O linkage reveal the existence of ferromagnetic interactions between Ni(II) centers via N3- ligands and antiferromagnetic interactions through the H-bonded moiety. The overall magnetic behavior of the system depends on the delicate balance between these two competing interactions. PMID- 16471489 TI - Theoretical study of the methyl transfer in guanidinoacetate methyltransferase. AB - The reaction mechanism of the guanidinoacetate methyltransferase (GAMT) enzyme has been investigated by means of density functional theory using the B3LYP hybrid functional. GAMT catalyzes the S-adenosyl-L-methionine (SAM)-dependent methylation of guanidinoacetate (GAA) to form creatine. A quantum chemical model was built on the basis of the recent crystal structure of GAMT complexed with S adenosylhomocysteine (SAH) and GAA. The methyl group transfer from SAM to N(E) of GAA is shown to occur concertedly with a proton transfer from NE to the neighboring OD1 of Asp134. Good agreement is found between the calculated barrier and the experimental rate. PMID- 16471490 TI - Time-resolved infrared dynamics of C-F bond activation by a tungsten metal carbonyl. AB - Chemical reactions that break, or activate, C-H and C-F are of tremendous synthetic interest. The intramolecular C-F bond activation of a tungsten carbonyl system has been studied by time-resolved infrared spectroscopy. The formation of solvent complexes and the final product are monitored using time-resolved infrared spectroscopy of the CO stretches. The rate of the reaction is shown to be limited by the formation of an intermolecular complex between the tungsten metal center and a solvent molecule. Comparison with DFT calculations shows that in the absence of solvent molecules the intramolecular complex with the tethered perfluorobenzene ring is energetically favorable, but is not the primary kinetic product because of the initial geometry of the complex. PMID- 16471491 TI - Selective oxidation of semiconducting single-wall carbon nanotubes by hydrogen peroxide. AB - Selective oxidation of single-wall carbon nanotubes (SWCNTs) by H2O2 was conducted at varying heating times and monitored by UV-vis-NIR spectroscopy. A major increase in the relative absorption intensity indicated a higher than 80% concentration of metallic SWCNTs in the final product. Here, it is suggested that semiconducting SWCNTs are more reactive than metallic SWCNTs because of hole doping by H2O2, resulting in faster oxidation. PMID- 16471492 TI - Solvent-driven formation of bolaamphiphilic vesicles. AB - We show that a spontaneous bending of single-layer bolaamphiphiles results from the frustration due to the competition between core-core and tail-solvent interactions. We find that spherical vesicles are stable under rather general assumptions on these interactions described within the Flory-Huggins theory. We consider also the deformation of the vesicles in an external magnetic field that has been recently experimentally observed. PMID- 16471494 TI - Shrinking nanowires by kinetically controlled electrooxidation. AB - Nanowires composed of antimony, gold, and bismuth telluride (Bi2Te3) were reduced in diameter by electrooxidation in aqueous solutions. When electrooxidation was carried out using low current densities (Jox < 150 microA cm(-2)), the mean wire diameter decreased in direct proportion to the oxidation time, as expected for a kinetically controlled process. Under these conditions, the diameter uniformity of nanowires remained constant as wires were shrunk from initial diameters of more than 120 nm to less than 40 nm, for Sb and Bi2Te3, and less than 60 nm for Au. Oxidized nanowires remained continuous for more than 100 microm. Electrooxidation at higher current densities rapidly introduced breaks into these nanowires. Electrochemical wire growth and shrinking by electrooxidation were integrated into a single electrochemical experiment that allowed the final mean diameter of nanowires to be specified with a precision of 5-10 nm. PMID- 16471493 TI - Electrostatic stitching in gel-phase supported phospholipid bilayers. AB - We show that mixing zwitterionic lipids with up to 20% mole % cationic lipids produces gel-phase supported lipid bilayers that are morphologically free of defects detectable using noncontact mode atomic force microscopy (AFM). This contrasts with the observation of massive defects when anionic lipid was added, and also when no charged lipid was added. Infrared measurements of headgroup orientation in the presence of cationic lipid show that the mean headgroup orientation changes only minimally when temperature is lowered from the fluid phase to the gel phase. This is consistent with a tentative explanation, based on simple electrostatic arguments, in which cationic lipids "stitch" the bilayers together. On the functional side, this study demonstrates a simple method by which to minimize defects in gel-supported phospholipid bilayers. PMID- 16471495 TI - Nanopatterning and nanocharge writing in layer-by-layer quinquethiophene/phthalocyanine ultrathin films. AB - Nanometer-scale patterning and charging in layer-by-layer (LbL) ultrathin films of quinquethiophene (5TN)/phthalocyanine (CuPS) provides a novel write-read device using a standard current-sensing atomic force microscopy (CS-AFM). The AFM height images showed dented or raised morphological features that could be selectively manipulated by changing the direction of the bias voltages. The conductivity was repeatedly changed between a conductive and insulating state, originating from an electrochemical charging-discharging effect. This was attributed to electrochemical ion transport and the residual mobile ions present in LbL films. Finally, the nanocharge pattern was written by CS-AFM and read out in a conductivity map image. PMID- 16471496 TI - Identification of color centers on MgO(001) thin films with scanning tunneling microscopy. AB - Localized electronic defects on the surface of a 4 monolayer (ML) thin MgO(001) film deposited on Ag(001) have been investigated by low-temperature scanning tunneling microscopy and spectroscopy. Depending on the location of the defect, we observe for the first time different defect energy levels in the band gap of MgO. The charge state of defects can be manipulated by interactions with the scanning tunneling microscope tip. Comparison with ground state energy levels of color centers on the MgO surface obtained from embedded cluster calculations corroborates the assignment of the defects to singly and doubly charged color centers. PMID- 16471497 TI - Pattern and feature designed growth of ZnO nanowire arrays for vertical devices. AB - An approach is demonstrated for growing aligned ZnO nanowire/nanorod arrays following a predesigned pattern and feature with controlled site, shape, distribution, and orientation. The technique relies on an integration of atomic force microscopy (AFM) nanomachining with catalytically activated vapor-liquid solid (VLS) growth. The pattern and growth locations are defined by the catalyst distribution created by AFM, and the orientation is determined by the epitaxial growth on a single-crystal substrate. The technique opens a variety of possibilities of using nanowire arrays as sensor arrays, piezoelectric antenna arrays, nanolasers, photonic band gap crystal, biosensors, and field emitters with controlled density, location, shape, and distribution according to a designed pattern and feature. PMID- 16471498 TI - Controlled precipitation of solubilized carbon nanotubes by delamination of DNA. AB - Polyaromatic molecules, such as rhodamine 6G and methylene blue, were found capable of precipitating DNA-solubilized single-walled carbon nanotubes from solution through a competitive binding mechanism whereby DNA is displaced from the nanotube surface, allowing the nanotubes to rebundle. This delamination of DNA also occurred when complementary oligonucleotides were used to hybridize specifically to the DNA coating on the nanotubes. These findings were expanded to include techniques for controlled desolubilization and to provide additional elucidation into the interaction of SWNTs and noncovalent solubilizing agents. PMID- 16471500 TI - Spontaneous formation of vesicles of diblock copolymer EO6BO11 in water: a SANS study. AB - Small angle neutron scattering (SANS) is used to study the structures formed in water by a diblock copolymer EO6BO11 (having 6 ethylene oxide, EO, and 11 butylene oxide, BO, units). The data show that polymer solutions over a broad concentration range (0.05-20 wt %) contain vesicular structures at room temperature. Interestingly, these vesicles could be formed without any external energy input, such as extrusion, which is commonly required for the formation of other block copolymer or lipid vesicles. The EO6BO11 vesicles are predominantly unilamellar at low polymer concentrations, whereas at higher polymer concentrations or temperatures there is a coexisting population of unilamellar and multilamellar vesicles. At a critical concentration and temperature, the vesicular structures fuse into lyotropic arrays of planar lamellar sheets. The findings from this study are in broad agreement with the work of Harris et al. (Langmuir, 2002, 18, 5337), who used electron microscopy to identify the vesicle phase in the same system. PMID- 16471499 TI - Growth of single-crystalline KNbO3 nanostructures. AB - This communication reports on the growth of highly uniform KNbO3 nanowires exhibiting a narrow diameter distribution around 60 nm and a length-to-width ratio up to 100. The nanowires were prepared by a hydrothermal route, which enables simple, gram-scale production. A systematic study of the synthesized nanowires in terms of the morphological and chemical characteristics was carried out by varying the temperature-pressure conditions and the composition of the starting mixture. The results indicate that highly uniform single-crystalline nanowires form within a narrow window of the ternary phase diagram of KOH-Nb2O5 H2O. PMID- 16471502 TI - Single-molecule microscopy studies of electric-field poling in chromophore polymer composite materials. AB - One strategy for increasing the efficiency of organic electrooptic devices based on chromophore-polymer composite materials is to improve chromophore ordering. In these materials, ordering is induced through the interaction of the chromophore dipole moment with an external electric field, applied at temperatures near the Tg of the polymer host, a process referred to as "poling". To provide insight into the molecular details of the poling process under conditions representative of device construction, the rotational dynamics of single 4-dicyano-methylene-2 methyl-6-(p-(dimethylamino)styryl)-4H-pyran (DCM) molecules in poly(methyl acrylate) at T = Tg + 11 degrees C in the presence and absence of an electric field are investigated using single-molecule confocal fluorescence microscopy. Single-molecule rotational dynamics are monitored through the time evolution of the fluorescence anisotropy. The anisotropy correlation function demonstrates nonexponential decay, with beta values derived from fits using the Kohlrausch Williams-Watts law ranging from 0.7 to 1 with beta(KWW) = 0.83. This observation is consistent with previous studies of molecular rotation dynamics in polymer melts and reflects the dynamical heterogeneity provided by the polymer host. The rotational dynamics of DCM are weakly perturbed in the presence of a 50 V/microm electric field, typical of the field strength employed in device construction. The expected perturbation of the rotational dynamics is determined and found to be consistent with the alignment potential created by the electric field relative to the amount of thermal energy available. The relevance of these findings with respect to current models of the poling process is discussed. This work demonstrates the utility of polarization-sensitive single-molecule microscopy in elucidating the details of molecular reorientation during poling. PMID- 16471501 TI - A salt-free zero-charged aqueous onion-phase enhances the solubility of fullerene C60 in water. AB - An onion-phase (multilamellar vesicular phase or Lalpha-phase) was prepared from salt-free zero-charged cationic and anionic (catanionic) surfactant mixtures of tetradecyltrimethylammonium hydroxide (TTAOH)/lauric acid (LA)/H2O. The H+ and OH counterions form water (TTAOH + LA --> TTAL + H2O), leaving the solution salt free. The onion-phase solution has novel properties including low conductivity, low osmotic pressure and unscreened electrostatic repulsions between cationic and anionic surfactants because of the absence of salt. The spherical multilamellar vesicles have an average 250 nm radius as measured by freeze-fracture transmission electron microscopy (FF-TEM) and the maximum interlayer distance, i.e., the thickness of the hydrophobic bilayer and the water layer, was calculated to be around 52 nm by small-angle X-ray scattering (SAXS). Extremely hydrophobic C60 fullerene can be solubilized in this salt-free zero-charged aqueous onion-phase. As a typical result, 0.588 mg.mL(-1) (approximately 0.82 mmol.L(-1)) C60 has been successfully solubilized into a 50 mmol.L(-1) catanionic surfactant onion-phase aqueous solution. The weight ratio of fullerene to TTAL is calculated to be around 1:40. Solubilization of C60 in the salt-free catanionic onion-phase solution was investigated by using different sample preparation routes, and a variety of techniques were used to characterize these vesicular systems with or without encapsulated C60. The onion-phase solution changed color from slightly bluish to yellow or brown after C60 was solubilized. 1H and 13C NMR measurements indicated that the C60 molecules are located in the hydrophobic layers, i.e., in the central positions [omega-CH3 and delta-(CH2)x] of the hydrophobic layers of the TTAL onion-phase. Salt-free zero-charged catanionic vesicular aqueous solutions are good candidates for enhancing the solubility of C60 in aqueous solutions and may broaden the functionality of fullerenes to new potential applications in biology, medicine, and materials. Hopefully, our method can also be extended to solubilize functionalized carbon nanotubes in aqueous solutions. PMID- 16471503 TI - Zeolite LTA Nanoparticles Prepared by Laser-Induced Fracture of Zeolite Microcrystals. AB - Zeolite LTA nanoparticles are prepared by laser-induced fragmentation of zeolite LTA microparticles using a pulsed laser. Zeolite nanoparticle formation is attributed to absorption of the laser at impurities or defects within the zeolite microcrystal generating thermoelastic stress that mechanically fractures the microparticle into smaller nanoparticle fragments. Experimentally, it is found that nanoparticles have a wide size and morphology distribution. Large nanoparticles (>200 nm) are typically irregularly shaped crystals of zeolite LTA, whereas small nanoparticles (<50 nm) tend to be spherical, dense, and amorphous, indicative of destruction of the original LTA crystal structure. Results of the fragmentation versus laser parameters show that shorter laser wavelengths are more efficient at producing zeolite nanoparticles, which is explained based on a larger cross section for optical absorption in the zeolite crystal. Increasing the laser energy density irradiating the sample was found to be a trade-off between increasing the amount of fragmentation and increasing the amount of structural damage to the zeolite crystal. It is suggested that in the presence of strongly absorbing defects, plasma formation is induced resulting in dramatically higher temperatures. On the basis of these results it is suggested the optimal laser processing conditions are 355 nm and 10 mJ/pulse laser energy for our LTA samples. PMID- 16471504 TI - Dynamics of p-nitroaniline molecules in micoporous aluminophosphate AlPO4-5 studied by solid-state NMR. AB - Dynamics of deuterated p-nitroaniline (pNA-d) molecules in the micropores of AlPO4-5 has been investigated by means of solid-state NMR. The adsorbed amounts of pNA-d were 5.0 and 10.1 mass % of the total mass. We have measured 13C magic angle-spinning (MAS) and 2H NMR spectra of the guest molecules and 31P and 27Al MAS NMR spectra of the host framework. The pNA-d molecules distribute rather inhomogeneously in the channel, and do not coordinate to Al strongly like H2O. The intermolecular hydrogen bonds are formed between a part of the guest molecules only when the loading level is high. The 2H NMR spectra are successfully analyzed, elucidating the orientation and the motion of the guest molecules. The molecular axis of pNA-d is inclined to the channel axis, and the molecular plane is perpendicular to the inner wall. The guest molecule jumps among 12 sites or 12 orientations. This motion is faster in the sample of 5.0 mass % than in the sample of 10.1 mass %, suggesting that the guest-guest interaction hinders the motion. The mean residence times of the molecules are estimated from the analysis of the 2H NMR spectra, which are affected by the size of the nanospace as well as the property of the adsorbed site. PMID- 16471505 TI - Dehydrated and Cs+-exchanged MFI zeolites: location and population of Cs+ from in situ diffraction data as a function of temperature and degree of exchange. AB - H-MFI type zeolitic materials of different Si/Al ratios have been completely or partially cesium-exchanged (cesium content ranging from 0.7 to 7.7 Cs/unit-cell (uc)). Examined with synchrotron X-ray powder diffractometry, an anhydrous sample with the Cs6.6H0.3Al6.9Si89.1O192 chemical composition revealed at ambient temperature the presence of five discrete Cs locations: Cs1 located in the channel intersection near a 10-ring window of the zigzag channel; Cs2 and Cs2', both located in the straight channel but 1.23 A apart; Cs3 and Cs3', both located in the zigzag channel and rather close to each other (2.51 A). The populations of the Cs species amounted to 2.61/0.81/1.85/0.86/0.47/uc for Cs1/2/2'/3/3', respectively. The continuous but multimodal nature of the C2 split site is well described by a joint-probability density function. The 10-ring of the straight channel in the framework is highly elliptical (epsilon = 1.218). The populations for the same sites were also determined at higher temperatures: 131, 237, 344, and 450 degrees C. At 450 degrees C, Cs2' has migrated toward the center of the channel intersection, and the site separation between Cs2 and Cs2' has lengthened to 2.23 A. Using a temperature-controlled laboratory X-ray diffractometer, similar studies were carried out on partially or almost totally Cs-exchanged samples from various sources with differing Cs contents. They show that over the 0.7 to 4 Cs/uc range all the individual Cs populations vary linearly as a function of total Cs/uc present. At higher total Cs/uc content (4 to approximately 7 Cs/uc) solely Cs1 continues to do so. For Cs2+Cs2' and Cs3+Cs3', the variation is almost linear over the whole concentration range. Computer simulations using a 6-exp-1 Buckingham-type atom-atom van der Waals interaction model yield six possible Cs sites in the actual Cs6.6MFI framework structure. Four of them lie very close to those determined from difference Fourier maps using the room temperature data. A fifth one is close to the Cs2' species after thermal migration at 450 degrees C, and the sixth one is close to the center of the channel intersection. However, this latter site is observed experimentally only in the case of hydrated CsMFI phases. In the anhydrous Cs6.6MFI phase at room temperature, the shortest Cs-framework oxygen distance is Cs3'-O25 = 3.08 A, and the next shortest distances are Cs1-O26 = 3.37, Cs2-O11 = 3.34, Cs2'-O22 = 3.47, and Cs3-O20 = 3.34 A. The framework T(Si,Al) sites most involved in these contacts are the T9, T11, T12, T10, and T3 sites. This implies that these sites are prime candidates for Si/Al substitution. PMID- 16471506 TI - Single wall carbon nanotube supports for portable direct methanol fuel cells. AB - Single-wall and multiwall carbon nanotubes are employed as carbon supports in direct methanol fuel cells (DMFC). The morphology and electrochemical activity of single-wall and multiwall carbon nanotubes obtained from different sources have been examined to probe the influence of carbon support on the overall performance of DMFC. The improved activity of the Pt-Ru catalyst dispersed on carbon nanotubes toward methanol oxidation is reflected as a shift in the onset potential and a lower charge transfer resistance at the electrode/electrolyte interface. The evaluation of carbon supports in a passive air breathing DMFC indicates that the observed power density depends on the nature and source of carbon nanostructures. The intrinsic property of the nanotubes, dispersion of the electrocatalyst and the electrochemically active surface area collectively influence the performance of the membrane electrode assembly (MEA). As compared to the commercial carbon black support, single wall carbon nanotubes when employed as the support for anchoring the electrocatalyst particles in the anode and cathode sides of MEA exhibited a approximately 30% enhancement in the power density of a single stack DMFC operating at 70 degrees C. PMID- 16471507 TI - Theoretical determination of multiple exchange couplings and magnetic susceptibility data in inorganic solids: the prototypical case of Cu2(OH)3NO3. AB - Density functional theory based on hybrid functionals and localized atomic type basis sets is employed to calculate the exchange couplings in the layered three dimensional compound Cu2(OH)3NO3. We assign accurate values to the six different in-plane exchange couplings. Interlayer exchange interactions through hydrogen bonds are also quantified. The calculated exchange coupling constants are then employed to perform quantum Monte Carlo simulations to yield magnetic susceptibility data, which compare successfully with experiments. Our approach sets the foundations of a viable methodology to extract reliable magnetic susceptibilities from density functional data. PMID- 16471508 TI - Vapor-transportation preparation and reversible lithium intercalation/deintercalation of alpha-MoO3 microrods. AB - We report on the preparation and electrochemical application of rechargeable lithium-ion batteries of alpha-MoO3 microrods. A simple and efficient vapor transportation approach was developed to yield large-scale alpha-MoO3 microrods. The as-prepared products were present in long, uniform, rodlike structures with a diameter of approximately 2 approximately 6 microm, and the proportion of the rod morphology was about 95% according to the analysis of scanning electron microscopy (SEM). The electrochemical lithium intercalation/deintercalation characteristic of the as-prepared microrods was investigated by cyclic voltammetry and a galvanostatic charge-discharge method. The results showed that the alpha-MoO3 microrods exhibited high capacity (225 mAh g(-1)) and excellent cycling reversibility, and are thus promising cathode candidates in advanced rechargeable lithium-ion batteries. The correlation between the specific structural features of the microrods and their superior electrode performance is discussed in detail, revealing that the unique rodlike structure plays an important role in optimizing the electrochemical performance of the electrode. PMID- 16471509 TI - A bis(p-sulfonatophenyl)phenylphosphine-based synthesis of hollow Pt nanospheres. AB - We report herewith the synthesis of hollow Pt nanospheres by using bis(p sulfonatophenyl)phenylphosphine to selectively remove the Ag cores of Ag-Pt core shell nanoparticles. Core-shell Ag-Pt nanoparticles were first obtained by the successive reduction method with a discontinuous Pt shell to allow the BSPP passage. Transmission electron microscopy imaging of the core-shell Ag-Pt nanoparticles before and after BSPP dissolution showed little changes in the particle size, indicating that the removal of the Ag cores had occurred isomorphously. The hollow Pt nanospheres, together with the predecessor Ag-Pt core-shell particles of the same size, were transferred from water to toluene and surface modified by dodecylamine in toluene. This allows the catalytic activities of solid and hollow Pt particles in room temperature methanol oxidation reaction to be compared under conditions of identical particle size and the same surface environment. The measured higher specific activity of the Pt hollow nanospheres could then be attributed unambiguously to the larger specific surface area prevalent in the porous hollow structure. PMID- 16471510 TI - High-yield synthesis of single-wall carbon nanotubes on MCM41 using catalytic chemical vapor deposition of acetylene. AB - High-quality single-wall carbon nanotubes (SWNTs) with narrow diameter distribution have been grown on Fe/Co-loaded MCM41 by using acetylene as the carbon source within a short reaction period, typically 10 min or less. The optimum temperature for SWNTs synthesis is 850 degrees C. Longer reaction time (i.e., 30 min) favors the formation of multiwall carbon nanotubes (MWNTs) and graphitic carbon. When the reaction time is reduced to less than 10 min, formation of MWNTs and graphitic carbon is greatly suppressed, and high-quality SWNTs dominates the yield. The surface of the as-grown SWNTs is found to be free from amorphous carbon, as observed from high-resolution transmission electron microscope (HRTEM) analysis. Raman spectral data show a G/D ratio above 10, indicating that the as-grown SWNTs have very few defects. Furthermore, radial breathing mode (RBM) analysis reveals that the diameter distribution of the current SWNTs is narrow and ranges from 0.64 to 1.36 nm. PMID- 16471511 TI - Ultrafast electron relaxation dynamics in coupled metal nanoparticles in aggregates. AB - We report the effect of aggregation in gold nanoparticles on their ultrafast electron-phonon relaxation dynamics measured by femtosecond transient absorption pump-probe spectroscopy. UV-visible extinction and transient absorption of the solution-stable aggregates of gold nanoparticles show a broad absorption in the 550-700-nm region in addition to the isolated gold nanoparticle plasmon resonance. This broad red-shifted absorption can be attributed to contributions from gold nanoparticle aggregates with different sizes and/or different fractal structures. The electron-phonon relaxation, reflected as a fast decay component of the transient bleach, is found to depend on the probe wavelength, suggesting that each wavelength interrogates one particular subset of the aggregates. As the probe wavelength is changed from 520 to 635 nm across the broad aggregate absorption, the rate of electron-phonon relaxation increases. The observed trend in the hot electron lifetimes can be explained on the basis of an increased overlap of the electron oscillation frequency with the phonon spectrum and enhanced interfacial electron scattering, with increasing extent of aggregation. The experimental results strongly suggest the presence of intercolloid electronic coupling within the nanoparticle aggregates, besides the well-known dipolar plasmon coupling. PMID- 16471512 TI - Size-dependent ultrafast electronic energy relaxation and enhanced fluorescence of copper nanoparticles. AB - The energy relaxation of the electrons in the conduction band of 12 and 30 nm diameter copper nanoparticles in colloidal solution was investigated using femtosecond time-resolved transient spectroscopy. Experimental results show that the hot electron energy relaxation is faster in 12 nm copper nanoparticles (0.37 ps) than that in 30 nm copper nanoparticles (0.51 ps), which is explained by the size-dependent electron-surface phonon coupling. Additional mechanisms involving trapping or energy transfer processes to the denser surface states (imperfection) in the smaller nanoparticles are needed to explain the relaxation rate in the 12 nm nanoparticles. The observed fluorescence quantum yield from these nanoparticles is found to be enhanced by roughly 5 orders of magnitude for the 30 nm nanoparticles and 4 orders of magnitude for the 12 nm nanoparticles (relative to bulk copper metal). The increase in the fluorescence quantum yield is attributed to the electromagnetic enhancement of the radiative recombination of the electrons in the s-p conduction band below the Fermi level with the holes in the d bands due to the strong surface plasmon oscillation in these nanoparticles. PMID- 16471513 TI - Gold nanorods to nanochains: mechanistic investigations on their longitudinal assembly using alpha,omega-alkanedithiols and interplasmon coupling. AB - Mechanistic investigations on the end-to-end assembly of Au nanorods to nanochains, in the presence of alpha,omega-alkanedithiols, were reported. A decrease in the longitudinal plasmon absorption was observed along with a concomitant formation of a new red-shifted band above a critical concentration of dithiol, which is attributed to the interplasmon coupling in assembled nanorods. However, no noticeable spectral changes were observed below the critical concentration, and the TEM studies indicate that the nanorods remain isolated and randomly distributed. This step is ascribed as an incubation step wherein one of the thiol groups of alpha,omega-alkanedithiol preferentially binds onto the edges of the nanorods, leaving the other thiol group free. Above the critical concentration, a chain up process proceeds through the interlocking of nanorods, initially to dimers and subsequently to oligomers, which results in longitudinal interplasmon coupling. The dimerization step follows second-order kinetics which deviates with time due to oligomerization. The rate constants for dimerization of nanorods possessing various dithiols and their energy of activation were determined. The large activation energy for the dimerization further confirms that the process is not diffusion but activation controlled. PMID- 16471514 TI - TiC nanocrystal formation from carburization of laser-grown Ti/O/C nanopowders for nanostructured ceramics. AB - Refractory carbide ceramics (TiC and ZrC) raise interest as promising materials for high-temperature applications such as structural materials for the future generation of nuclear reactors. In this context, nanostructured ceramics are expected to exhibit improved thermomechanical properties as well as better behavior under irradiation when compared to conventional materials. It is therefore necessary to synthesize carbide nanocrystals of such materials to elaborate the ceramics. We report here the formation study of TiC nanocrystals through the direct carburization of Ti/O/C nanopowders grown by laser pyrolysis. A spray of titanium tetraisopropoxide was laser pyrolyzed with ethylene as the sensitizer, leading to Ti/O/C nanopowders with various C contents controlled by the synthesis conditions. Annealing treatments performed on these nanopowders under an inert atmosphere without any C addition enabled the formation of TiC grains through the carburization of the oxide phase by free C incorporated during the synthesis. The powders were characterized by X-ray diffraction, scanning electron microscopy, and transmission electron microscopy. The final TiC grain size was about 80 nm, and the grains were monocrystalline. The influence of the free C content on the grain growth during the annealing step, together with its effects on the densification of the ceramics after sintering by high-pressure flash sintering, was examined. A 93% densification was finally achieved. PMID- 16471515 TI - Raman spectroscopy of free-standing individual semiconducting single-wall carbon nanotubes. AB - The radial breathing modes and tangential modes have been systematically measured on a large number of individual semiconducting single-wall carbon nanotubes (thin bundles) suspended between plots (free-standing single-wall carbon nanotubes). The strong intensity of the Raman spectra ensures the precision of the experimentally determined line shapes and frequencies of these modes. The diameter dependence of the frequencies of the tangential modes was measured. This dependence is discussed in relation with recent calculations. The present data confirm/contradict some previous interpretations. PMID- 16471516 TI - The role of the central atom in structure and reactivity of polyoxometalates with adjacent d-electron metal sites. computational and experimental studies of gamma [(Xn+O4)RuIII2(OH)2(MFM)10O32](8-n)-) for MFM = Mo and W, and X = AlIII, SiIV, PV, and SVI. AB - The role of the central atom X in the structure and reactivity of di-Ru substituted gamma-Keggin polyoxometalates (POMs), gamma [(Xn+O4)RuIII2(OH)2(MFM)10O32](8-n)-), where MFM = Mo and W, and X = AlIII, SiIV, PV, and SVI., was computationally investigated. It was shown that for both MFM = Mo and W the nature of X is crucial in determining the lower lying electronic states of the polyoxoanions, which in turn likely significantly impacts their reactivity. For the electropositive X = AlIII, the ground state is a low-spin state, while for the more electronegative X = SVI the ground state is a high-spin state. In other words, the heteroatom X can be an "internal switch" for defining the ground electronic states of the gamma-M2-Keggin POMs. The obtained trends, in general, are less pronounced for MFM = Mo than for W. On the basis of the comparison of the calculated energy gaps between low-spin and high-spin states of polytungstates and polymolybdates, we predict that the gamma-M2-Keggin polytungstates could be more reactive than their polymolybdate analogues. For purposes of experimental verification the computationally predicted and evaluated polytungstate gamma-[(SiO4)RuIII2(OH)2(OH2)2W10O32]4- was prepared and characterized. PMID- 16471517 TI - Synthesis and structural study on MnO2 nanosheet material by X-ray absorption spectroscopic technique. AB - MnO2 nanosheet with acetylene black composite material has been synthesized from layered K0.45MnO2 powder. The electrochemical lithiation reaction of nanosheet composite material proceeds in a different manner from that of the parent material, layered K0.45MnO2 powder. To elucidate the origin of the changes in discharge profile, the electronic and local structures for the nanosheet composites and its parent and protonated material have been investigated by Mn K edge and O K-edge X-ray absorption spectroscopy (XAS). The results showed that local and electronic structure around Mn ions does not vary during nanosheet formation, while significant changes in electronic structure around oxide ions were observed. Accordingly, it is suggested that the difference observed in discharge profile is due to the electronic structural change induced by nanosheet formation. PMID- 16471518 TI - Synthesis and structure characterization of chromium oxide prepared by solid thermal decomposition reaction. AB - Mesoporous chromium oxide (Cr2O3) nanocrystals were first synthesized by the thermal decomposition reaction of Cr(NO3)3.9H2O using citric acid monohydrate (CA) as the mesoporous template agent. The texture and chemistry of chromium oxide nanocrystals were characterized by N2 adsorption-desorption isotherms, FTIR, X-ray diffraction (XRD), UV-vis, and thermoanalytical methods. It was shown that the hydrate water and CA are the crucial factors in influencing the formation of mesoporous Cr2O3 nanocrystals in the mixture system. The decomposition of CA results in the formation of a mesoporous structure with wormlike pores. The hydrate water of the mixture provides surface hydroxyls that act as binders, making the nanocrystals aggregate. The pore structures and phases of chromium oxide are affected by the ratio of precursor-to-CA, thermal temperature, and time. PMID- 16471519 TI - Synthesis and evolution of PbS nanocrystals through a surfactant-assisted solvothermal route. AB - We present a surfactant-assisted solvothermal approach for the controllable synthesis of a PbS nanocrystal at low temperature (85 degrees C). Nanotubes (400 nm in length with an outer diameter of 30 nm), bundle-like long nanorods (about 5 15 mum long and an average diameter of 100 nm), nanowires (5-20 mum in length and with a diameter of 20-50 nm), short nanorods (100-300 nm in length and an axial ratio of 5-10), nanoparticles (25 nm in width with an aspect ratio of 2), and nanocubes (a short axis length of 10 nm and a long axis length of 15 nm) were successfully prepared and characterized by transmission electron microscopy, scanning electron microscopy, and powder X-ray diffraction pattern. A series of experimental results indicated that several experimental factors, such as AOT concentration, ratio of [water]/[surfactant], reaction time, and ratio of the reagents, play key roles in the final morphologies of PbS. Possible formation mechanisms of PbS nanorods and nanotubes were proposed. PMID- 16471520 TI - Altering associations in aqueous solutions of a hydrophobically modified alginate in the presence of beta-cyclodextrin monomers. AB - The formation of associative networks in semidilute aqueous solutions of hydrophobically modified alginate (HM-alginate) is dependent on intermolecular hydrophobic interactions. Addition of beta-cyclodextrin (beta-CD) monomers to the system provides decoupling of these associations via inclusion complex formation with the polymer hydrophobic tails. This results in a dramatic decrease in the viscoelastic response of the system and a more extended local structure of the polymer chains, as shown by small-angle neutron scattering (SANS) measurements. The zero-shear viscosity decreases about an order of magnitude when the beta-CD concentration is increased from 0 to 12 mm. The lifetime of the associative network decreases strongly with increasing levels of beta-CD addition. These findings clearly demonstrate that the hydrophobic association effect is efficiently reduced as the amount of beta-CD is increased. In the framework of drug delivery, this effect may be useful to improve the release of therapeutic molecules that can be entrapped in the polymer matrix. PMID- 16471521 TI - Influence of graphite addition on the reactivity of Ti powder with H2 under ball milling. AB - The effect of graphite addition on the mechanism of hydrogen uptake by titanium during mechanochemical activation in hydrogen flow was studied using kinetic, structural, microscopic, and spectroscopic techniques. As was found, already a small graphite admixture of about 0.5 wt % changed the kinetics of mechanically induced H2 sorption and significantly stimulated Ti-H2 interaction. Two new types of occupation sites available for hydrogen were observed, which are characterized by low H2 desorption temperatures: about 650 and 750 K instead of 1000 K. PMID- 16471522 TI - One-pot synthesis of hollow Au3Cu1 spherical-like and biomineral botallackite Cu2(OH)3Cl flowerlike architectures exhibiting antimicrobial activity. AB - A new form of Au3Cu1 hollow nanostructure was prepared by the reaction of Cu nanoparticles with HAuCl4. Following a course of aging, the biomineral botallackite Cu2(OH)3Cl nanoflowers were developed with the aid of Au3Cu1 hollow nanostructures at room temperature. It was proposed that the hollow nanospheres could serve as active centers for heterogeneous nucleation and mediated a mineralization process. Scanning electron microscopy and high-resolution transmission electron microscopy indicated that the nanoflowers are three dimensional in appearance with a range of 500 nm-- to 1 microm in size and made of several nanopetals with about 25 nm in thickness. In addition, we found that the shape separation could be achieved by using cationic cetyltrimethylammonium bromide to filter the different morphology spherical- and flowerlike structures due to the negative charge of hollow nanospheres. Both hollow nanospheres and nanoflowers presented antimicrobial activity toward Streptococcus aureus with MIC50 at 39.6 and 127.2 microg/mL, respectively. PMID- 16471523 TI - Kinetics of receptor directed assembly of multisegment nanowires. AB - We demonstrate the receptor directed end-to-end assembly of multisegment Au/Ni/Au nanowires under agitation in ethanol. The gold end-segments were functionalized with biotin-terminated thiol thereby restricting aggregation to end-to-end attachment via an avidin linkage. On mixing biotin-terminated nanowires with avidin-terminated nanowires, the average chain length is shown to increase linearly with time. The rate constant was independent of the nanowire concentration. Kinetic Monte Carlo simulations were used to model the self assembly process, and we show that the directed end-to-end assembly of nanowires is similar to the polycondensation of linear polymers. PMID- 16471524 TI - Strain energies due to nonplanar distortion of fullerenes and their dependence on structural motifs. AB - Accurate strain energies due to nonplanar distortion of 114 isolated pentagon rule (IPR) fullerenes with 60-102 carbon atoms have been calculated based on B3LYP/6-31G(d) optimized structures. The calculated values of strain energy due to nonplanar distortion (Enp) are reproduced by three simple schemes based upon counts of 8, 16, and 30 distinct structural motifs composed of hexagons and pentagons. Using C180 (Ih) and CN (Ih) (N is very large) as test molecules, the intrinsic limitations of the motif model based on six-membered rings (6-MRs) as the central unit have been discussed. On the basis of the relationship between the contributions of motifs to Enp and the number of five-membered rings (5-MRs) in motifs, we found that IPR fullerenes with dispersed 5-MRs present smaller nonplanar distortions. PMID- 16471525 TI - SiO2 nanowires growing on hexagonally arranged circular patterns surrounded by TiO2 films. AB - An effective approach has been demonstrated for the synthesis of novel composite architectures, SiO2 nanowires (NWs) growing on hexagonally arranged circular patterns surrounded by TiO2 films on Si substrate. First, a solution-dipping template strategy is used to create TiO2 films with hexagonally arranged pores on Au-coated Si substrate, resulting in hexagonally arranged circular patterns of catalysts surrounded by TiO2 films. Then the patterned catalysts guide the growth of SiO2 NWs with the original TiO2 films preserved, realizing the composite structures. Such composite architectures combine the photoluminescence (PL) properties of the two components, and also present more favorable PL property, laying a foundation for future advanced nano-optoelectronic devices. PMID- 16471526 TI - High-pressure studies of optical dephasing in polymer glasses. AB - The effect of high pressure on the optical dephasing of chromophores in organic polymers at low temperature is evaluated within the stochastic sudden jump two level-system (TLS) model. The approximations within the "standard" TLS model cannot account for the observed pressure dependence of the pure dephasing rate without ad hoc assumptions about changes in the TLS density of states. However, the photon echo model of Geva and Skinner for disordered systems can be used to model pressure-dependent optical dephasing results for a variety of doped polymer systems without assuming changes in the TLS density of states. The relative importance of pressure-induced changes in TLS density, chromophore-TLS coupling, and TLS-phonon coupling is evaluated by fitting experimental high-pressure photon echo results to the TLS model. PMID- 16471527 TI - Characterization of surface composition of platinum and ruthenium nanoalloys dispersed on active carbon. AB - Supported samples of 8 wt % monometallic Pt/C and Ru/C, as well as 12 wt % bimetallic Pt50Ru50/C, were prepared by the method of incipient wetness impregnation. Impregnated samples were subsequently reduced by hydrogen and then oxidized in air at different To temperatures. TEM and XRD examinations indicated that metal crystallites were finely dispersed with a diameter of dM < or = 3 nm on the reduced samples. Reductive behavior of the oxidized samples by hydrogen was pursued with the technique of temperature programmed reduction (TPR). The temperature of the reduction peaks (Tr) noticed in the TPR profiles varied with the metal composition of catalysts and To temperature of oxidation. At To = 300 K, oxidation was confined to the surface layer of metallic crystallites. As a result, Pts O (with a peak at Tr = 230 K) or PtsO2 (Tr = 250 K) was formed on monometallic Pt/C while RusO2 (Tr approximately 380 K) was formed on Ru/C. A reductive peak with Tr = 250 K was found from the bimetallic sample from Pt50Ru50/C oxidized at To = 300 K. The reductive peak suggests bimetallic crystallites were dispersed with cherry type structure, with Pt exposed at the surface and Ru in the core. On increasing the To temperature of oxidation treatment to 370 K and higher, Tr peaks between 270 and 350 K were gradually noticed on the oxidized bimetallic sample. Peaks in this Tr region are assigned to reduction of the oxidized alloy surface (AsO). Evidently, a segregation of Ru to the surface of the bimetallic crystallites is indicated upon oxidation at To > 380 K. PMID- 16471528 TI - An in-depth study of supported In2O3 catalysts for the selective catalytic reduction of NOx: the influence of the oxide support. AB - The influence of the oxide support (i.e., Al2O3, Nb2O5, SiO2, and TiO2,) on the surface properties, reduction and oxidation properties, acid-base properties, and catalytic activity of supported indium oxide catalysts has been investigated by temperature-programmed reduction/oxidation, thermogravimetry coupled to differential scanning calorimetry, ammonia and sulfur dioxide adsorption calorimetry, and reduction of NOx by ethene in highly oxygen-rich atmosphere. Two series of In2O3-containing catalysts at low (approximately 3 wt %) and at theoretical geometric monolayer (from 20 to 40 wt %) In2O3 content were prepared and their properties were compared with unsupported In2O3 material. Supports able to disperse the In2O3 aggregates with high In stabilization gave rise to active catalytic systems. Among the studied oxide supports, Al2O3 and, to a lower extent, TiO2 were found to be the best supports for obtaining active de-NOx catalysts. PMID- 16471529 TI - On the difficulties of predicting the adsorption of volatile organic compounds at low pressures in microporous solid: the example of ethyl benzene. AB - Adsorption isotherms of toluene and ethyl benzene, at 25 degrees C and 40 degrees C, were determined in two microporous activated carbons and one zeolite. Significant differences were found in the adsorption behavior, at low pressures, between the two vapors on the same adsorbent material. The quantities of adsorbed ethyl benzene at 25 degrees C, in the low-pressure region, were lower than what was observed at 40 degrees C in all the studied adsorbents, contrary to what was found for toluene. This fact was not related to kinetic effects at the two temperatures nor to vapor swelling of the adsorbents structure. Also, there was no molecular sieving since at high pressures the toluene and ethyl benzene occupied the same adsorption space. The differences found in the ethyl benzene adsorption at the two temperatures pose difficulties in the analysis of the adsorption data and, therefore, in the prediction of results. This is discussed in the analysis of the results with the application of the Dubinin-Astakhov equation and in the estimation of the isosteric heats of adsorption. The adsorption potentials of two possible ethyl benzene conformations were estimated for the adsorption in the pores of activated carbon from the Horvath and Kawazoe model, and the values compared with those found experimentally. The results were interpreted in terms of the ethyl benzene conformation effects when the molecule is confined in pores that are about the same size of one of the conformations. PMID- 16471531 TI - Identification and measurements of strong Bronsted acid site in ultrastable Y (USY) zeolite. AB - By using the IRMS-TPD method in which IR (infrared) and MS (mass spectroscopy) worked together, acid sites of USY (ultrastable Y) zeolite were studied. A new band of OH playing a role of Bronsted acid was clearly detected on Na2H2 ethylenediaminetetraacetic acid (EDTA)-treated USY at 3595 cm(-1) during an elevation in temperature after the adsorption of ammonia. MS-measured TPD (temperature-programmed desorption) of NH3 and IR-measured TPD of the NH4+ cation coincided well to show that this zeolite consisted of the Bronsted acid sites. The MS-TPD profile at higher temperatures corresponded to the IR-TPD of the 3595 cm(-1) band, and therefore, this OH was identified as a strong acid site. From comparison between IR-TPD of OH and MS-TPD, numbers of three kinds of Bronsted OH (i.e., those in super and sodalite cages of a Y zeolite structure) and created strong Bronsted acid site were quantified. On the other hand, strength of the Bronsted acid site DeltaH was determined individually by a simulation method, where the corrected IR-TPD of OH was simulated based on the proposed equation. Thus, a new strong Bronsted acid site was identified in the EDTA-treated USY, and the amount and strength was measured quantitatively. PMID- 16471530 TI - Effects of surface chemical composition on the early growth stages of alpha sexithienyl films on silicon oxide substrates. AB - In organic field effect transistors, charge transport is confined to a narrow region next to the organic/dielectric interface. It is thus extremely important to determine the morphology and the molecular arrangement of the organic films at their early growth stages. On a substrate of technological interest, such as thermally grown silicon oxide, it has been recently found that alpha-sexithienyl aggregates made of flat-lying molecules can simultaneously nucleate besides islands made of molecules standing vertical. In this paper, we investigate the effects due to variations in surface chemical composition on alpha-sexithienyl ultrathin film formation. Flat-lying molecules are no longer detected when Si-OH groups present at the surface are chemically removed but also when the Si-OH or Si-H group density is maximized. This gives evidence that variations in the surface chemical composition can largely affect the nucleation and growth processes of organic/dielectric interfaces. We hypothesize that isolated OH groups can interact with alpha-sexithienyl molecules and anchor them down flat with respect to the surface. PMID- 16471532 TI - Formation of the C-S-H Layer during early hydration of tricalcium silicate grains with different sizes. AB - Portland cement is a mixture of solid phases which all react with water. Tricalcium silicate (Ca3SiO5) is its main component and is often used in model systems to study cement hydration. It is generally recognized that setting and hardening of cement are due to the formation, by a dissolution-precipitation process, of a calcium silicate hydrate (C-S-H) on anhydrous grains during Ca3SiO5 hydration. The purpose of this paper is to study the effect of Ca3SiO5 particle size on the nucleation-growth process of C-S-H. An experimental study of the rate of hydration by using different grain sizes under controlled conditions has been performed. The experimental data have been compared with results obtained by numerical simulation based on a particles-aggregation model. The thickness of the C-S-H layer when the reaction becomes limited by diffusion has been then estimated. PMID- 16471533 TI - Optical properties of the poly(N-benzylaniline) thin film. AB - The optical properties of the poly(N-benzylaniline) thin film were investigated by optical characterization. The optical constants such as refractive index and dielectric constant were determined from the transmittance and reflectance spectra of the film. The refractive index dispersion was analyzed by the Wemple DiDomenico model. The n(infinity) values changed from 6.37 to 5.71 and these values did not show any certain trend with annealing temperatures. The average oscillator parameter So value, which is the strength of the individual dipole oscillator, was found to be in the range of 1.15 x 10(13) to 1.03 x 10(13) m(-2). The optical band was determined from the direct optical transitions in K space. The optical band Eg of the film decreases from 2.089 to 2.046 eV with increasing annealing temperatures while the Urbach energy Eu called the width of localized states in the optical band gap increases from 0.544 to 0.598 eV. Consequently, the optical constants and optical band gap of the poly(N-benzylaniline) change with the annealing temperatures. PMID- 16471535 TI - A new approach to the spectral analysis of liquid membrane oscillators by Gabor transformation. AB - Liquid membrane oscillators very frequently have an irregular oscillatory behavior. Fourier transformation cannot be used for these nonstationary oscillations to establish their power spectra. This important point seems to be overlooked in the field of chemical oscillators. A new approach is presented here based on Gabor transformation allowing one to obtain power spectra of any kind of oscillations that can be met experimentally. The proposed Gabor analysis is applied to a liquid membrane oscillator containing a cationic surfactant. It was found that the power spectra are strongly influenced by the presence of various added substances. PMID- 16471534 TI - Photocatalytic oxidation of propylene with molecular oxygen over highly dispersed titanium, vanadium, and chromium oxides on silica. AB - Photocatalytic oxidation of propylene with molecular oxygen at room temperature was investigated over various silica-supported metal oxides with low loading. The photocatalytic active site is assumed to be the isolated tetrahedrally coordinated metal oxides in the ligand-to-metal charge-transferred state, such as (Mdelta- -OLdelta+). Photocatalytic epoxidation of propylene into propylene oxide was promoted over silica-supported V and Ti oxides at steady state. Over silica supported Cr oxide, the propylene oxide formation rate was remarkably decreased with the time course in the reaction. The oxidation state and the coordination environment of the supported Ti, V, and Cr oxide species were determined by diffuse reflectance UV-vis spectroscopy (DRS) and electron spin resonance (ESR). During the photocatalytic oxidation, the oxidation state of the Ti4+ species was not varied. On the other hand, the V5+ species was partially reduced to V4+ and the Cr6+ species was successively reduced to Cr5+ and Cr3+. An isotopic tracer study of the C3H6-18O2 reaction suggests the difference of the active oxygen species between TiO2/SiO2 and V2O5/SiO2. The active oxygen species on TiO2/SiO2 is derived from molecular oxygen. On the other hand, the photogenerated products on V2O5/SiO2 incorporate the lattice oxygen of the surface metal oxide species. It is suggested that the kinds of terminal ligand (hydroxyl or oxo) of the tetrahedrally coordinated metal oxides on silica decide the active oxygen species in the photocatalytic oxidation. A photoinduced hole center on the monohydroxyl (SiO)3Ti-OH species activates molecular oxygen that reacts with propylene. In the case of the monooxo (SiO)3V=O and dioxo (SiO)2Cr=O2 species, the photoactivated lattice oxygen (OL-) directly reacts with propylene. PMID- 16471536 TI - Three-stage multilayer formation kinetics during adsorption of an anionic fluorinated surfactant onto germanium. 1. Concentration effect. AB - The adsorption of tetraethylammonium perfluorooctylsulfonate (TEA-FOS) from aqueous solution onto hydroxylated germanium is studied using in situ polarized attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR). The adsorption is monitored at a series of bulk solution concentrations spanning from well below to above the critical micelle concentration (CMC; 1.0 mM). The kinetics of adsorption is followed by monitoring the intensity of the fluorocarbon bands. The orientation of the fluorocarbon director with respect to the germanium surface is determined by circular dichroism measurements of CF2 stretching bands. At bulk concentrations ranging from 10% of the CMC to at least 500% of the CMC, the adsorption occurs in an unusual sequence of three stages. Initially, rapid adsorption occurs within 200 min, leading to coverage of a monolayer or less. A long period of slow adsorption follows, during which we hypothesize that surfactant molecules form clusters, some of which serve as nuclei for multilayer growth. This stage concludes suddenly with an acceleration in the rate of adsorption, which eventually leads to multilayer formation. Because this is an anionic surfactant adsorbing onto a negatively charged surface at pH 6, the tetraethylammonium ions must mediate the interactions between the surfactant headgroups and the surface. The dichroism measurements show that TEA FOS is initially oriented randomly or somewhat parallel to the surface, but over time adopts an orientation somewhat normal to the surface. This behavior is consistent with initial adsorption at isolated sites, followed by aggregation into isotropic admicelles, and finally growth into flattened admicelles. The sudden onset of accelerated adsorption can be explained either by autoaccelerating adsorption or nucleation and growth of a hydrophobic multilayer structure. PMID- 16471537 TI - Formation of the surface NO during N2O interaction at low temperature with iron containing ZSM-5. AB - Interaction of N2O at low temperatures (473-603 K) with Fe-ZSM-5 zeolites (Fe, 0.01-2.1 wt %) activated by steaming and/or thermal treatment in He at 1323 K was studied by the transient response method and temperature-programmed desorption (TPD). Diffuse reflectance infrared fourier transform spectroscopy (DRIFTS) of NO adsorbed at room temperature as a probe molecule indicated heterogeneity of surface Fe(II) sites. The most intensive bands were found at 1878 and 1891 cm( 1), characteristic of two types mononitrosyl species assigned to Fe2+(NO) involved in bi- and oligonuclear species. Fast loading of atomic oxygen from N2O on the surface and slower formation of adsorbed NO species were observed. The initial rate of adsorbed NO formation was linearly dependent on the concentration of active Fe sites assigned to bi- and oligonuclear species, evolving oxygen in the TPD at around 630-670 K. The maximal coverage of a zeolite surface by NO was estimated from the TPD of NO at approximately 700 K. This allowed the simulation of the dynamics of the adsorbed NO formation at 523 K, which was consistent with the experiments. The adsorbed NO facilitated the atomic oxygen recombination/desorption, the rate determining step during N2O decomposition to O2 and N2, taking place at temperatures > or =563 K. PMID- 16471538 TI - 29Si NMR studies of zeolite precursor solutions. AB - Solution 29Si NMR spectroscopy results of zeolite precursor solutions of composition 1 SiO2:4 C2H5OH:0.36/n R+n[OH-]n:20 H2O are reported. This work employs isotopically enriched 29Si materials to aid in spectral interpretation. Using both 1D and 2D methods, spectra of solutions containing tetrapropylammonium hydroxide are wholly consistent with the existing silicate chemistry literature and indicate that the majority of the species are high-symmetry silicate clusters previously observed in aqueous solutions. The results are inconsistent with the nanoblock or nanoslab model proposed by Kirschhock and co-workers. Mixtures containing the 4,4'-trimethylene-bis(1,1'-dimethylpiperidinium) dihydroxide cation were also studied. These mixtures have similar speciation to the TPA solutions, although the relative populations of the species are different. Preliminary variable temperature 29Si NMR of these mixtures shows that the exchange properties of the high-symmetry silicate species, most notably the tetrahedral tetramer, depend on the organocation identity. PMID- 16471539 TI - Generation of atomistic models of periodic mesoporous silica by kinetic Monte Carlo simulation of the synthesis of the material. AB - We have developed a molecular simulation method for the generation of realistic atomic-level models for periodic mesoporous silicas. Using simplified interaction potentials and simplified representations of the templating micelles, the simulation follows the reaction path of the hydrothermal synthesis and calcination of the silica material in a kinetic Monte Carlo (kMC) simulation. The only input to the simulation is the geometry of the micelle and the number of silicic acid monomers at the beginning of the synthesis. We simulated the adsorption properties of the PMS models using Grand Canonical Monte Carlo simulation. With use of MCM-41 materials of different pore sizes as a prototype material, experimental and simulated adsorption isotherms for nitrogen, ethane, and carbon dioxide were compared, showing good agreement between simulation and experiment. PMID- 16471540 TI - Structure and adsorption of a hard-core multi-Yukawa fluid confined in a slitlike pore: grand canonical Monte Carlo simulation and density functional study. AB - Because of the increasing interest in studying the phenomenon exhibited by charge stabilized colloidal suspensions in confining geometry, we present a density functional theory (DFT) for a hard-core multi-Yukawa fluid. The excess Helmholtz free-energy functional is constructed by using the modified fundamental measure theory and Rosenfeld's perturbative method, in which the bulk direct correlation function is obtained from the first-order mean spherical approximation. To validate the established theory, grand canonical ensemble Monte Carlo (GCMC) simulations are carried out to determine the density profiles and surface excesses of multi-Yukawa fluid in a slitlike pore. Comparisons of the theoretical results with the GCMC data suggest that the present DFT gives very accurate density profiles and surface excesses of multi-Yukawa fluid in the slitlike pore as well as the radial distribution functions of the bulk fluid. Both the DFT and the GCMC simulations predict the depletion of the multi-Yukawa fluid near a nonattractive wall, while the mean-field theory fails to describe this depletion in some cases. Because the simple form of the direct correlation function is used, the present DFT is computationally as efficient as the mean-field theory, but reproduces the simulation data much better than the mean-field theory. PMID- 16471542 TI - Exploring three-dimensional nanosystems with Raman spectroscopy: methylene blue adsorbed on thiol and sulfur monolayers on gold. AB - Resonant Raman and surface-enhanced Raman scattering (SERS) spectroscopies, complemented with scanning tunnel microscopy and electrochemical techniques, have been used to obtain information about the amount and spatial distribution of methylene blue (MB) molecules immobilized on sulfur and four ultrathin molecular alkanethiolate films self-assembled on Au(111) and rough Au electrodes. The intensity of the Raman signals allow one to estimate the amount of immobilized MB at different organic films, whereas the decrease in the SERS intensity as a function of distance for the rough Au electrodes is used to locate the average position of the MB species with respect to the Au substrate. We found that significant amounts of cationic MB species are able to diffuse into methyl terminated thiols, but they are stopped at the outer plane of the self-assembled monolayer (SAM) by negatively charged carboxylate groups. The relative shift of C N stretching Raman modes indicates that the binding of MB to S is different from that found for MB on thiols. Most of the molecules immobilized on methyl- and carboxylate-terminated thiols are electrochemically inactive, suggesting that strong coupling between the Au electrode and the MB molecules is needed for charge transfer. Our results are consistent with a small population of electrochemically active MB species very close to the Au surface that reach this position driven by their lipophilic (hydrophobic) character through defects at SAMs. PMID- 16471541 TI - Adsorption structures of water in NaX studied by DRIFT spectroscopy and neutron powder diffraction. AB - Diffuse reflectance infrared Fourier transform spectroscopic (DRIFTS) measurements (4000-1500 cm(-1)) and the results of neutron powder diffraction have been combined to study the structure of adsorption complexes of water in a NaX zeolite at different water loadings (25, 48, 72, and 120 water molecules per unit cell, respectively). Sharp bands corresponding to non-hydrogen-bonded OH groups of water molecules and broad associate bands due to hydrogen-bonded molecules are observed in the DRIFT spectra. We observe a remarkable downshift of the high-frequency associate band in a narrow temperature interval when the water amount decreases from 120 to 72 molecules per unit cell, which could signify some kind of "phase transition" for the water inside the zeolite cavities. Neutron powder diffraction results show that water molecules are predominantly localized in or near the 12-ring windows. Water molecules with hydrogen-bonded and non hydrogen-bonded OH groups were found, in agreement with the observation of sharp and broad bands in the DRIFT spectra. We find strong evidence for the formation of cyclic hexamers of water molecules localized in the 12-ring windows, which are further stabilized by hydrogen bonds to framework oxygen atoms. PMID- 16471543 TI - Electrode independent chemoresistive response for cobalt phthalocyanine in the space charge limited conductivity regime. AB - The electrical properties of 50 nm thick metallophthalocyanine films, prepared by organic molecular beam epitaxy (OMBE) on interdigitated electrodes, were studied with DC current-voltage measurements and impedance spectroscopy. The transition from Ohmic behavior at low voltages to space-charge-limited conductivity (SCLC) at higher voltages depends on the metal electrode (Pt, Pd, and Au), but does not correlate with the work function of the electrode. Impedance spectroscopy studies show the coexistence of low- and high-frequency traps in the thin film devices, and the contribution of low-frequency traps associated with Ohmic behavior diminishes at higher bias. Although device resistances are strongly influenced by the electrode material, and vary by a factor of over 300, the relative chemical sensor responses on exposure to dimethyl methylphosphonate (DMMP), methanol, water, or toluene vapors are similar for CoPc on Pt, Pd, and Au electrodes when these devices are operated in the SCLC regime at room temperature. When the devices are operated at voltages where the low-frequency interfacial traps are filled, the sensor response to analyte becomes uniform and reliable regardless of the specific interfacial electrode contact. PMID- 16471544 TI - DFT study of a weakly pi-bonded C2H4 on oxygen-covered Ag(100). AB - The coadsorption of ethylene, C2H4, and atomic oxygen on Ag(100) was studied using density-functional theory. As for the adsorption of oxygen alone, the on surface hollow sites are predicted to be the most stable adsorption sites at low coverage (< or =1/2 ML). Above this coverage, mixed on-surface + subsurface oxygen configurations become more stable. The binding of ethylene to the clean Ag(100) is weak and little affected by oxygen when it is adsorbed on-surface. On the other hand, we find that the adsorption energy of C2H4 may increase considerably when oxygen is adsorbed into subsurface sites. Our results indicate that the increased reactivity of surface Ag atoms is because of their decreased coordination due to the push out effect of oxygen underneath, more than to their oxidation. PMID- 16471545 TI - The electrochemical corrosion of bulk nanocrystalline ingot iron in acidic sulfate solution. AB - The corrosion properties of bulk nanocrystalline ingot iron (BNII) fabricated from conventional polycrystalline ingot iron (CPII) by severe rolling were investigated by means of immersion test, potentiodynamic polarization (PDP), electrochemical impedance spectroscopy (EIS) tests, and scanning electron microscopy (SEM) observation. These experimental results indicate that BNII possesses excellent corrosion resistance in comparison with CPII in acidic sulfate solution at room temperature. It may mainly result from different surface microstructures between CPII and BNII. However, the corrosion resistance of nanocrystalline materials is usually degraded because of their metastable microstructure nature, and the residual stress in nanocrystalline materials also can result in degradation of corrosion resistance according to the traditional point of view. PMID- 16471546 TI - Synthesis of palladium nanoparticles by sonochemical reduction of palladium(II) nitrate in aqueous solution. AB - The sonochemical synthesis of stable palladium nanoparticles has been achieved by ultrasonic irradiation of palladium(II) nitrate solution. The starting solutions were prepared by the addition of different concentrations of palladium(II) nitrate in ethylene glycol and poly(vinylpyrrolidone) (PVP). The resulting mixtures were irradiated with ultrasonic 50 kHz waves in a glass vessel for 180 min. The UV-visible absorption spectroscopy and pH measurements revealed that the reduction of Pd(II) to metallic Pd has been successfully achieved and that the obtained suspensions have a long shelf life. The protective effect of PVP was studied using Fourier transform infrared (FT-IR) spectroscopy. It has been found that, in the presence of ethylene glycol, the stabilization of the nanoparticles results from the adsorption of the PVP chain on the palladium particle surface via the coordination of the PVP carbonyl group to the palladium atoms. The effect of the initial Pd(II) concentration on the Pd nanoparticle morphology has been investigated by transmission electron microscopy. It has been shown that the increase of the Pd(II)/PVP molar ratio from 0.13 x 10(-3) to 0.53 x 10(-3) decreases the number of palladium nanoparticles with a slight increase in particle size. For the highest Pd(II)/PVP value, 0.53 x 10(-3), the reduction reaction leads to the unexpected smallest nanoparticles in the form of aggregates. PMID- 16471548 TI - Combined theoretical and FTIR spectroscopic studies on hydrogen adsorption on the zeolites Na-FER and K-FER. AB - The interaction between molecular hydrogen and the alkali-metal-exchanged zeolites Na-FER and K-FER at a low temperature was investigated by combining variable-temperature infrared spectroscopy and theoretical calculations by using a periodic DFT model. The experimentally determined values of standard adsorption enthalpy, DeltaH degrees , were -6.0 (+/-0.8) and -3.5 (+/-0.8) kJ mol(-1) for Na FER and K-FER, respectively. These results were found to be in agreement with corresponding DeltaH degrees values obtained from calculations on the periodic model. Two types of alkali-metal cation sites in FER were found: channel intersection sites and channel wall sites. Calculations showed a similar interaction energy for both site types, and similar structures of adsorption complexes. Up to two dihydrogen molecules can be physisorbed on the alkali-metal cation located on the intersection of two channels, while only one H2 molecule is physisorbed on the cation at the channel wall site. The adsorption enthalpies of H2 on alkali-metal-exchanged FER are significantly smaller than those found previously for the MFI-type zeolites Na-ZSM-5 and K-ZSM-5, which is likely due to a difference in the alkali-metal cation coordination in the two zeolite frameworks. PMID- 16471547 TI - Organic pigment particles coated with titania via sol-gel process. AB - This paper presented a novel method for the organic pigment coated with titania to improve the weatherability and dispersion ability in waterborne system. The organic pigment was first orderly adsorbed by two kinds of electrolyte: poly(sodium 4-styrenesulfonate) (PSS) and poly(diallyldimethylammonium chloride) (PDADMAC), then coated by titania via sol-gel process from titanium n-butoxide (TBOT). The effects of the numbers of polyelectrolyte layer, water content, and TBOT content on the morphology, particle size, surface element composition, porosity and pore size, thermal stability, and UV shielding property of the organic pigment were systematically investigated. It was found that only two layers of electrolyte adsorption and one-step coating of titania could obviously enhance the UV shielding property even thermal stability of the organic pigment. The thickness of the titania layer could be easily tailored by TBOT content. PMID- 16471549 TI - Friction and adhesion between C60 single crystal surfaces and AFM tips: effects of the orientational phase transition. AB - We have investigated the nanotribological properties of C60 single crystal (111) and (100) surfaces around its orientational order-disorder phase transition temperature, approximately 260 K, by atomic force microscopy and frictional force microscopy (AFM/FFM) in high vacuum. Results show that for both surfaces across the phase transition temperature, the friction force and the adhesive force between a C60 coated AFM tip and the C60 crystal surfaces exhibit discontinuous behavior. The friction force within the applied external load range in the low temperature phase is significantly larger than that in the high temperature phase, with no obvious change in the slope of the friction force curves (the friction coefficient) in the low and high temperature phases. The abrupt change in friction was found to be caused mainly by the abrupt change in adhesion, which, in turn, can be qualitatively understood through changes in the van der Waals interaction and the short-range Coulomb interaction associated with the structural changes across the phase transition. Compared to most other degrees of freedom, the rotation of C60 molecules was found to have little effect on friction and is an ineffective energy dissipation channel. PMID- 16471550 TI - Femtosecond fluorescence dynamics of porphyrin in solution and solid films: the effects of aggregation and interfacial electron transfer between porphyrin and TiO2. AB - The excited-state relaxation dynamics of a synthetic porphyrin, ZnCAPEBPP, in solution, coated on a glass substrate as solid films, mixed with PMMA and coated on a glass substrate as solid films, and sensitized on nanocrystalline TiO2 films were investigated by using femtosecond fluorescence up-conversion spectroscopy with excitation in the Soret band, S2. We found that the S2--> S1 electronic relaxation of ZnCAPEBPP in solution and on PMMA films occurs in 910 and 690 fs, respectively, but it becomes extremely rapid, <100 fs, in solid films and TiO2 films due to formation of porphyrin aggregates. When probed in the S1 state of porphyrin, the fluorescence transients of the solid films show a biphasic kinetic feature with the rapid and slow components decaying in 1.9-2.4 and 19-26 ps, respectively. The transients in ZnCAPEBPP/TiO2 films also feature two relaxation processes but they occur on different time scales, 100-300 fs and 0.8-4.1 ps, and contain a small offset. According to the variation of relaxation period as a function of molecular density on a TiO2 surface, we assigned the femtosecond component of the TiO2 films as due to indirect interfacial electron transfer through a phenylethynyl bridge attached to one of four meso positions of the porphyrin ring, and the picosecond component arising from intermolecular energy transfer among porphyrins. The observed variation of aggregate-induced relaxation periods between solid and TiO2 films is due mainly to aggregation of two types: J type aggregation is dominant in the former case whereas H-type aggregation prevails in the latter case. PMID- 16471551 TI - Structure-activity relationship in HC-SCR of NOx by TEM, O2-chemisorption, and EDXS study of Ag/Al2O3. AB - Ag/alumina catalysts with different silver loading (1.28-6 wt %) for lean NO reduction activity were prepared by impregnation and the incipient wetness method. Complementary HRTEM, HAADF, O2-chemisorption, and EDXS studies were applied to investigate the dependence between silver particle size and catalytic activities of the prepared materials. The catalyst with the lowest silver loading (1.28 wt %) was found to be the most active catalyst in terms of reacted NO molecules per mole of silver. On the basis of the HRTEM, HAADF, and O2 chemisorption studies it could be concluded that the mean particle size or particle size distribution of the samples alone could not explain the big difference in the activities. EDXS analyses showed on the other hand that all of the samples were very heterogeneous in terms of particle size distribution, e.g., including both small and very big particles. Furthermore, both metallic silver and mainly hexagonal silver oxide (Ag2O) were found to be present in the samples. Despite the valuable information provided by ex situ characterization of the prepared samples, it needs to be emphasized that establishing a structure reactivity relationship for this type of catalyst requires in situ characterization. PMID- 16471552 TI - In situ studies of the active sites for the water gas shift reaction over Cu-CeO2 catalysts: complex interaction between metallic copper and oxygen vacancies of ceria. AB - New information about the active sites for the water gas shift (WGS) reaction over Cu-CeO2 systems was obtained using in-situ, time-resolved X-ray diffraction (TR-XRD), X-ray absorption spectroscopy (TR-XAS, Cu K and Ce L3 edges), and infrared spectroscopy (DRIFTS). Cu-CeO2 nanoparticles prepared by a novel reversed microemulsion method (doped Ce1-xCuxO2 sample) and an impregnation method (impregnated CuOx/CeO2 sample) were studied. The results from all of the samples indicate that both metallic copper and oxygen vacancies in ceria were involved in the generation of active sites for the WGS reaction. Evidence was found for a synergistic Cu-Ovacancy interaction. This interaction enhances the chemical activity of Cu, and the presence of Cu facilitates the formation of O vacancies in ceria under reaction conditions. Water dissociation occurred on the Ovacancy sites or the Cu-Ovacancy interface. No significant amounts of formate were formed on the catalysts during the WGS reaction. The presence of strongly bound carbonates is an important factor for the deactivation of the catalysts at high temperatures. This work identifies for the first time the active sites for the WGS reaction on Cu-CeO2 catalysts and illustrates the importance of in situ structural studies for heterogeneous catalytic reactions. PMID- 16471553 TI - Chemical activity of photoinduced Ti3+ centers in titanium oxide gels. AB - We report on the chemical activity of trapped electrons in wet titanium oxide gels. These electrons are generated under the band gap irradiation of gels in the spectral range between 3.25 and 4.4 eV and stored as Ti3+ centers that absorb in the visible. Chemical processes in photoirradiated gels are generally similar to those earlier reported in TiO2 colloids; however, peculiarities exist. In particular, a high internal surface of gels strongly enhances interface reactions. Measurements of UV-visible absorption kinetics allow conclusions to be made about extremely high available traps concentration and the activity of all trapped electrons toward nitrate and nitrite anion reduction according to a heterogeneous photocatalytic mechanism. PMID- 16471554 TI - Controlling the energy transfer in dipole chains. AB - Processing digital signals on the molecular scale is of great interest. In this paper, we discuss the control of pulselike energy propagation through one dimensional arrays of dipoles. Three systems are explored. In the first system, a chain of coaxial dipoles is gated by two control dipoles. Changing the orientation of these control dipoles lets us control the transfer of energy in the chain. In the other two systems, the chain-branch system and the two-branch system, two chains are used as an input and the propagation of energy is controlled by sending one or two signals toward the junction. Both systems can operate as a logical AND port. Their geometrical configurations are key to a well defined control and operation of the AND port. PMID- 16471555 TI - Understanding the influence of polymorphism on phonon spectra: lattice dynamics calculations and terahertz spectroscopy of carbamazepine. AB - Rigid molecule atomistic lattice dynamics calculations have been performed to predict the phonon spectra of the four polymorphs of carbamazepine, and these calculations predict that there should be differences in the spectra of all four forms. Terahertz spectra have been measured for forms I and III, and there are clearly different features between polymorphs' spectra, that are accentuated at low temperature. While carbamazepine adopts the same hydrogen bonded dimers in all of its known polymorphs, the calculations show that differences in packing arrangements of the dimers lead to changes in the frequency ranges for each type of hydrogen bond vibration, giving a physical explanation to the observed differences between the spectra. Although the agreement between calculated and observed spectra does not allow a definitive characterization of the spectra, it is possible to make tentative assignments of many of the observed features in the terahertz region for the simpler form III; we can only make some tentative assignments of specific modes in the more complex spectrum of form I. While harmonic rigid molecule lattice dynamics shows promise for understanding the differences in spectra between polymorphs of organic molecules, discrepancies between observed and calculated spectra suggest areas of improvement in the computational methods for more accurate modeling of the dynamics in molecular organic crystals. PMID- 16471556 TI - Dynamics of the amorphous and crystalline alpha-, gamma-phases of indomethacin. AB - Crystallization processes in indomethacin can be observed below Tg leading to different forms depending on the thermal treatment: a rapid and deep quench below Tg leads to the metastable alpha-phase and a slow cooling close to Tg gives rise to the stable gamma-phase. To understand this atypical behavior, we have studied the molecular mobility of the amorphous and crystalline forms of indomethacin by dielectric relaxation and 1H NMR spectroscopy. Two relaxations were detected in the glassy state obtained from the deeply quenched liquid. One, also present in the gamma-phase, was attributed to local rotations. The other one, of very low amplitude, was attributed to the Johari-Goldstein relaxation. The results allowed to discuss the relationship between these two relaxation processes and the crystallization properties of amorphous indomethacin. PMID- 16471557 TI - Self-consistent-field analysis of the micellization of carboxy-modified poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) triblock copolymers. AB - The micellization properties of carboxy-modified Pluronics P85 (poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) triblock copolymers) are investigated by means of a molecularly realistic self-consistent field theory. We consider the, so-called, carboxylic acid end-standing P85 (CAE 85) case where the carboxylic group is located at the end of both PEO parts and the carboxylic acid center-standing P85 (CAC-85) case where each of the carboxylic group presents between the PEO and PPO blocks. The micellization of these copolymers depends on the pH, the added electrolyte concentration phis, and the temperature. It is shown that the aggregation number (Nagg) decreases, whereas the critical micellization concentration (CMC) increases with pH. For the case of increasing phis, the Nagg increases and the CMC decreases. The critical micellization temperature (CMT) and cloud point temperature (CPT) increase with pH at low phis and decrease at increasing phis. The changing from CAE-85 to CAC 85 leads to increasing CMC and CMT, but lower CPT. PMID- 16471558 TI - Can the speed of sound be used for detecting critical states of fluid mixtures? AB - The phenomenology of sound speeds in fluid mixtures is examined near and across critical lines. Using literature data for binary and ternary mixtures, it is shown that the ultrasound speed along an isotherm-isopleth passes through a minimum value in the form of an angular (or V-shaped) point at critical states. The relation between critical and pseudo-critical coordinates is discussed. For nonazeotropic fixed-composition fluid mixtures, pseudo-critical temperatures and pressures are found to be lower than the corresponding critical temperatures and pressures. The analysis shows that unstable pseudo-critical states cannot be detected using acoustic methods. The thermodynamic link between sound speeds and isochoric heat capacities is formulated and discussed in terms of p-Vm-T derivatives capable of being calculated using cubic equations of state. Based on the Griffiths-Wheeler theory of critical phenomena, a new specific link between critical sound speeds and critical isochoric heat capacities is deduced in terms of the rate of change of critical pressures and critical temperatures along the p T projection of the critical locus of binary fluid mixtures. It is shown that the latter link can be used to obtain estimates of critical isochoric heat capacities from the experimental determination of critical speeds of sound. The applicability domain of the new link does not include binary systems at compositions along the critical line for which the rate of change in pressure with temperature changes sign. The new equation is combined with thermodynamic data to provide approximate numerical estimates for the speed of sound in two mixtures of carbon dioxide and ethane at different temperatures along their critical isochores. A clear decrease in the sound speed is found at critical points. A similar behavior is suggested by available critical heat capacity data for several binary fluid mixtures. Using an acoustic technique, the critical temperature and pressure were determined for three different mixtures of methane and propane, and compared with literature data obtained using conventional methods. It is concluded that acoustic-based techniques are reliable to determine, for the most part, critical surfaces of fluid mixtures. The remaining few cases where the present analysis cannot be applied could be tested by the thermodynamic calculation of critical sound speeds using crossover equations of state in conjunction with experimentally determined critical isochoric heat capacities. PMID- 16471559 TI - Evidences of nonideal mixing in poly(ethylene glycol)/organic solvent mixtures by Brillouin scattering. AB - The concentration dependence of the hypersonic properties of solutions of poly(ethylene glycol) of mean molecular mass 600 g/mol (PEG600) in benzene and toluene has been investigated by Brillouin scattering. The two solvents are very similar in structure and chemical properties, but while benzene is nonpolar, toluene possess a modest dipole. In both solvents a high-frequency relaxation process has been observed at high concentrations which has been assigned to conformational rearrangements of the polymeric chains, triggered by reorientation of the side groups. In both cases, the concentration dependence of the adiabatic compressibility deviates significantly from linearity, indicating the existence of nonideal mixing phenomena driven by aggregation processes taking place in the systems. However, there is no temperature dependence for solutions of PEG600 in benzene; on the contrary, the results obtained for solutions of PEG600 in toluene are noticeably dependent on the temperature. The comparison of the experimental data with the results of previous experiments on similar systems allows a general picture for weakly interacting mixtures of hydrogen-bonded systems and organic solvents to be developed. In particular, in the presence of a nonpolar solvent molecule the local structure of the mixture is dominated by solute self association processes and any resulting solute-solvent correlation is barely induced by excluded volume effects. At high enough dilution the self-aggregation of solute molecules produces a variety of new local topologies that cannot be observed in bulk solute, and as a consequence, the concentration evolution of the system is too rich to be described in terms of a linear combination of a few components over the whole concentration range. The situation seems to be simpler for the polar toluene solvent molecules, where a three-component model seems able to fit the experimental concentration dependence of the hypersonic velocity. This result is interpreted to imply that the interaction between the solvent dipoles and the active sites of the solute produces a relatively stable heterocoordination, while the relevance of self-association is partially reduced. PMID- 16471560 TI - Temperature- and pressure-dependence of the outer-sphere reorganization free energy for electron transfer reactions: a continuum approach. AB - The outer-sphere reorganization free energy for electron-transfer reactions in polar solvents and its variations with temperature and pressure are studied in the dielectric continuum framework by extending the recent fluctuating cavity description [J. Chem. Phys. 2005, 123, 014504]. The diabatic free energies are obtained as a function of three variables, i.e., radii of two spherical cavities for the donor and acceptor moieties of an electron-transfer complex and a solvent coordinate that gauges an arbitrary configuration of solvent orientational polarization. Equilibrium cavities relevant to the reactant and product states are determined via the variational principle. This incorporates cavity size readjustment accompanying electron transfer and related electrostrictive effects. Another important consequence of the variational determination of equilibrium cavities is that their size depends on thermodynamic conditions. The application of the theoretical formulation presented here to electron self-exchange shows that in contrast to the prediction of the standard Marcus theory, the solvent reorganization free energy decreases with temperature. This is in excellent accord with a recent experiment on a mixed valence dinuclear iron complex in acetonitrile [J. Phys. Chem. A 1999, 103, 7888]. It is also found that electrostriction makes a significant contribution to outer-sphere reorganization. Model calculations for the dinuclear iron complex system show that about 25-30% of the total solvent reorganization free energy arises from cavity size changes, while solvent repolarization is responsible for the rest. PMID- 16471561 TI - Molecular dynamics-based approach to study the anisotropic self-diffusion of molecules in porous materials with multiple cage types: application to H2 in losod. AB - The anisotropic self-diffusion of molecular hydrogen in the multiple cage clathrasil losod (LOS) is modeled by means of molecular dynamics (MD) simulations of up to 1 micros for the temperature range 900-1200 K while treating the framework as fully flexible. The LOS diffusion tensor is calculated employing an analytical method based on hopping rates. The diffusion in the c-direction of the unit cell is found to be approximately two times more rapid than in the a- and the b-directions, a characteristic of importance for the application of LOS as a membrane. The overall diffusion is based on five different hop types for which the individual hopping rates and diffusion barriers are calculated separately. We show explicitly that the shape and volume of the cages have a significant effect on the hopping rates and further that even small deformations of the circular Si6O6 apertures have a large influence on the energetic barrier for hydrogen diffusion. Compared to the single cage clathrasils dodecasil 3C (MTN) and sodalite (SOD), LOS has a lower diffusion rate. However, from a technical point of view this rate (at 573 K) is still fast enough for LOS to be interesting as a size-selective membrane or as a hydrogen-adsorption medium. PMID- 16471562 TI - Self-diffusion of supercooled o-terphenyl near the glass transition temperature. AB - Self-diffusion coefficients for the low molecular weight glass former o-terphenyl have been measured near Tg by isothermally desorbing thin film bilayers of deuterio and protio o-terphenyl in a vacuum chamber. We observe translational diffusion that is about 100 times faster at Tg + 3 K than the Stokes-Einstein prediction. Predictions from random first order transition theory and a dynamic facilitation approach are in reasonable agreement with our results; in these approaches, enhanced translational diffusion is associated with spatially heterogeneous dynamics. Self-diffusion controls crystallization in o-terphenyl for most of the supercooled liquid regime, but at temperatures below Tg + 10 K, the reported crystallization rate increases suddenly while the self-diffusion coefficient does not. This work and previous work on trisnaphthylbenzene both find a self-diffusion-controlled crystal growth regime and an enhancement in self diffusion near Tg, suggesting that these phenomena are general characteristics of fragile low molecular weight glass formers. We discuss the width of the relaxation time distributions of o-terphenyl and trisnaphthylbenzene as they relate to the observation of enhanced translational diffusion. PMID- 16471563 TI - Use of ultrafast dispersed pump-dump-probe and pump-repump-probe spectroscopies to explore the light-induced dynamics of peridinin in solution. AB - Optical pump-induced dynamics of the highly asymmetric carotenoid peridinin in methanol was studied by dispersed pump-probe, pump-dump-probe, and pump-repump probe transient absorption spectroscopy in the visible region. Dispersed pump probe measurements show that the decay of the initially excited S2 state populates two excited states, the S1 and the intramolecular charge-transfer (ICT) state, at a ratio determined by the excitation wavelength. The ensuing spectral evolution occurs on the time scale of a few picoseconds and suggests the equilibration of these states. Dumping the stimulated emission of the ICT state with an additional 800-nm pulse after 400- and 530-nm excitation preferentially removes the ICT state contribution from the broad excited-state absorption, allowing for its spectral characterization. At the same time, an unrelaxed ground state species, which has a subpicosecond lifetime, is populated. The application of the 800-nm pulse at early times, when the S2 state is still populated, led to direct generation of the peridinin cation, observed for the first time in a transient absorption experiment. The excited and ground electronic states manifold of peridinin has been reconstructed using target analysis; this approach combined with the measured multipulse spectroscopic data allows us to estimate the spectra and time scales of the corresponding transient states. PMID- 16471564 TI - Redox processes of cytochrome c immobilized on solid supported polyelectrolyte multilayers. AB - The heme protein cytochrome c (Cyt-c), immobilized on polyelectrolyte multilayers on a silver electrode, was studied by stationary and time-resolved surface enhanced resonance Raman (SERR) spectroscopy to probe the redox site structure and the mechanism and dynamics of the potential-dependent interfacial processes. The layers were built up by sequential adsorption of polycations (poly[ethylene imine] (PEI); polyallylamine hydrochloride (PAH)) and polyanions (poly[styrene sulfonate] (PSS)). All multilayers terminated by PSS electrostatically bind Cyt c. On PEI/PSS coatings, Cyt-c is peripherally bound and fully redox-active. Due to the interfacial potential drop, the apparent redox potential is lowered by 40 mV compared to that in solution. The rate constant for the heterogeneous electron transfer (ET) of ca. 0.1 s(-1) is consistent with electron tunneling through largely ordered PEI/PSS layers. ET is coupled to a reversible conformational transition of Cyt-c that involves a change of the coordination pattern of the heme. Additional (PAH/PSS) double layers cause a broadening of the redox transition and a drastic negative shift of the redox potential, which is attributed to the formation of PSS/Cyt-c complexes. It is concluded that Cyt-c can effectively compete with PAH for binding of PSS, resulting in a rearrangement of the layered structure and a penetration of the PSS-bound Cyt-c into the PAH/PSS double layers. This conclusion is consistent with SERR intensity and quartz microbalance measurements. ET was found to be overpotential-independent and faster than that for PEI/PSS coatings, which is interpreted in terms of specific PSS/Cyt-c complexes serving as gates for the heterogeneous ET. PMID- 16471565 TI - Fe vibrational spectroscopy of myoglobin and cytochrome f. AB - The Fe vibrational density of states (VDOS) has been determined for the heme proteins deoxymyoglobin, metmyoglobin, and cytochrome f in the oxidized and reduced states, using nuclear resonance vibrational spectroscopy (NRVS). For cytochrome f in particular, the NRVS spectrum is compared with multiwavelength resonance Raman spectra to identify those Raman modes with significant Fe displacement. Modes not seen by Raman due to optical selection rules appear in the NRVS spectrum. The mean Fe force constant extracted from the VDOS illustrates how Fe dynamics varies among these four monoheme proteins, and is correlated with oxidation and spin state trends seen in model heme compounds. The protein's contribution to Fe motion is dominant at low frequencies, where coupling to the backbone tightly constrains Fe displacements in cytochrome f, in contrast to enhanced heme flexibility in myoglobin. PMID- 16471566 TI - Experimental/theoretical electrostatic properties of a styrylquinoline-type HIV-1 integrase inhibitor and its progenitors. AB - We have established that polyhydroxylated styrylquinolines are potent inhibitors of HIV-1 integrase (IN). Among them, we have identified (E)-8-hydroxy-2-[2-(4,5 dihydroxy-3-methoxyphenyl)-ethenyl]-7-quinolinecarboxylic acid (1) as a promising lead. Previous molecular dynamics simulations and docking procedures have shown that the inhibitory activity involves one or two metal cations (Mg2+), which are present in the vicinity of the active center of the enzyme. However, such methods are generally based on a force-field approach and still remain not as reliable as ab initio calculations with extended basis sets on the whole system. To go further in this area, the aim of the present study was to evaluate the predictive ability of the electron density and electrostatic properties in the structure activity relationships of this class of HIV-1 antiviral drugs. The electron properties of the two chemical progenitors of 1 were derived from both high resolution X-ray diffraction experiments and ab initio calculations. The twinning phenomenon and solvent disorder were observed during the crystal structure determination of 1. Molecule 1 exhibits a planar s-trans conformation, and a zwitterionic form in the crystalline state is obtained. This geometry was used for ab initio calculations, which were performed to characterize the electronic properties of 1. The electron densities, electrostatic potentials, and atomic charges of 1 and its progenitors are here compared and analyzed. The experimental and theoretical deformation density bond peaks are very comparable for the two progenitors. However, the experimental electrostatic potential is strongly affected by the crystal field and cannot straightforwardly be used as a predictive index. The weak difference in the theoretical electron densities between 1 and its progenitors reveals that each component of 1 conserves its intrinsic properties, an assumption reinforced by a 13C NMR study. This is also shown through an excellent correlation of the atomic charges for the common fragments. The electrostatic potential minima in zwitterionic and nonzwitterionic forms of 1 are discussed in relation with the localization of possible metal chelation sites. PMID- 16471567 TI - Molecular dynamics simulations of large integral membrane proteins with an implicit membrane model. AB - The heterogeneous dielectric generalized Born (HDGB) methodology is an the extension of the GBMV model for the simulation of integral membrane proteins with an implicit membrane environment. Three large integral membrane proteins, the bacteriorhodopsin monomer and trimer and the BtuCD protein, were simulated with the HDGB model in order to evaluate how well thermodynamic and dynamic properties are reproduced. Effects of the truncation of electrostatic interactions were examined. For all proteins, the HDGB model was able to generate stable trajectories that remained close to the starting experimental structures, in excellent agreement with explicit membrane simulations. Dynamic properties evaluated through a comparison of B-factors are also in good agreement with experiment and explicit membrane simulations. However, overall flexibility was slightly underestimated with the HDGB model unless a very large electrostatic cutoff is employed. Results with the HDGB model are further compared with equivalent simulations in implicit aqueous solvent, demonstrating that the membrane environment leads to more realistic simulations. PMID- 16471568 TI - Origin of the pKa perturbation of N-terminal cysteine in alpha- and 3(10) helices: a computational DFT study. AB - It is well documented that helices in proteins can decrease the pKa of residues located at the N-terminus, but the real nature of this perturbation remains unclear. In the present work, the origin of the effect of 3(10)- and alpha polyalanine helices on the pKa of an N-terminal cysteine residue is examined in gas phase as well as in aqueous solution by means of density functional theory. In a systematic study of the helix dipole, the proton affinity (PA), and the pKa of the N-terminal cysteine, in relation to both the helix length and the strength of the hydrogen bonds between the helix backbone amides and the Sgamma of the N terminal cysteine, a direct relation between the terminal hydrogen bonds and the pKa perturbation is revealed. PMID- 16471569 TI - Extended weak bonding interactions in DNA: pi-stacking (base-base), base backbone, and backbone-backbone interactions. AB - We report on several weak interactions in nucleic acids, which, collectively, can make a nonnegligible contribution to the structure and stability of these molecules. Fragments of DNA were obtained from previously determined accurate experimental geometries and their electron density distributions calculated using density functional theory (DFT). The electron densities were analyzed topologically according to the quantum theory of atoms in molecules (AIM). A web of closed-shell bonding interactions is shown to connect neighboring base pairs in base-pair duplexes and in dinuleotide steps. This bonding underlies the well known pi-stacking interaction between adjacent nucleic acid bases and is characterized topologically for the first time. Two less widely appreciated modes of weak closed-shell interactions in nucleic acids are also described: (i) interactions between atoms in the bases and atoms belonging to the backbone (base backbone) and (ii) interactions among atoms within the backbone itself (backbone backbone). These interactions include hydrogen bonding, dihydrogen bonding, hydrogen-hydrogen bonding, and several other weak closed-shell X-Y interactions (X, Y = O, N, C). While each individual interaction is very weak and typically accompanied by perhaps 0.5-3 kcal/mol, the sum total of these interactions is postulated to play a role in stabilizing the structure of nucleic acids. The Watson-and-Crick hydrogen bonding is also characterized in detail at the experimental geometries as a prelude to the discussion of the modes of interactions listed in the title. PMID- 16471570 TI - 31P NMR chemical shifts in hypervalent oxyphosphoranes and polymeric orthophosphates. AB - We report the first quantum chemical investigation of the solid- and solution state 31P NMR chemical shifts in models for phosphoryl transfer enzyme reaction intermediates and in polymeric inorganic phosphates. The 31P NMR chemical shifts of five- and six-coordinate oxyphosphoranes containing a variety of substitutions at phosphorus, as well as four-coordinate polymeric orthophosphates and four coordinate phosphonates, are predicted with a slope of 1.00 and an R2= 0.993 (N = 34), corresponding to a 3.8 ppm (or 2.1%) error over the entire 178.3 ppm experimental chemical shift range, using Hartree-Fock methods. For the oxyphosphoranes, we used either experimental crystallographic structures or, when these were not available, fully geometry optimized molecular structures. For the four-coordinate phosphonates we used X-ray structures together with charge field perturbation, to represent lattice interactions. For the three-dimensional orthophosphates (BPO4, AlPO4, GaPO4 we again used X-ray structures, but for these inorganic systems we employed a self-consistent charge field perturbation approach on large clusters, to deduce peripheral atom charges. For pentaoxyphosphoranes, the solvent effect on 31P NMR chemical shieldings was found to be very small (<0.5 ppm). The 31P NMR chemical shielding tensors in the pentaoxyphosphoranes were in most cases found to be close to axially symmetric and were dominated by changes in the shielding tensor components in the equatorial plane (sigma22 and sigma33). The isotropic shifts were highly correlated (R2= 0.923) with phosphorus natural bonding orbital charges, with the larger charges being associated with shorter axial P-O bond lengths and, hence, more shielding. Overall, these results should facilitate the use of 31P NMR techniques in investigating the structures of more complex systems, such as phosphoryl transfer enzymes, as well as in investigating other, complex oxide structures. PMID- 16471571 TI - Sequential deprotonation of meso-(p-hydroxyphenyl)porphyrins in DMF: from hyperporphyrins to sodium porphyrin complexes. AB - Sequential deprotonations of meso-(p-hydroxyphenyl)porphyrins (p-OHTPPH2) in DMF + H2O (V/V = 1:1) mixture have been verified to result in the appearance of hyperporphyrin spectra. However, when the deprotonations of these p-OHTPPH2 are carried out in DMF, the spectral changes differ considerably from those in the mixture mentioned above. At low [OH-], the optical spectra in the visible region are still considered to have characteristics of hyperporphyrin spectra. Further deprotonation at much higher basicity makes the optical spectra form three-banded spectra similar to those in the acidic solution. To clarify the molecular origins of these changes, UV-vis, resonance Raman (RR), proton nuclear magnetic resonance (1H NMR) experiments are carried out. Our data give evidence that p-OHTPPH2 in DMF can be further deprotonated of pyrrolic-H by higher concentrated NaOH, due to an aprotic medium like DMF effectively weakening the basicity of the porphyrin relative to that of the NaOH, and coordinates with two sodium ions (except the sodium ions that interact with the peripherial phenoxide anions) to form the sodium complexes of p-OHTPPH2 (Na2P, to lay a strong emphasis on the sodium ions that coordinate with the central nitrogen atom), which can be regarded as the porphyrin anions being perturbed by the sodium cations due to their highly ionic character. The negative centers generated by deprotonation of pyrrolic-H and phenolic-H are not thoroughly delocalized between the substituents and the porphyrin ring. Thus the negative centers generated by deprotonation of pyrrolic H only act as electron-donating groups on the porphyrin pi system, and the negative charges of the phenoxide anion are also mainly localized on the peripheral substituents. As a result, the porphyrin pi orbitals cross over the phenoxide anion pi orbital and turn into HOMOs, which turns hyperporphyrin spectra of deprotonated phenolic-H of p-OHTPPH2 into three-banded spectra of regular metalloporphyrins. PMID- 16471572 TI - Formation and structural anomaly of the metastable phases in an immiscible Ag-Mo system studied by ion beam mixing and molecular dynamics simulation. AB - For the equilibrium immiscible Ag-Mo system characterized by a large positive heat of formation, the nanosized Ag-Mo multilayered samples are designed and prepared to include sufficient interfacial free energy to elevate their initial energetic states to be higher than that of either the amorphous phase or solid solution and then subject to 200 keV xenon ion irradiation. The results show that a uniform amorphous alloy can be obtained within a composition range, at least, from 25 to 88 atom % of Mo. Interestingly, in the intermediate stage of ion irradiation, a bcc phase, an amorphous phase, and an order (bcc)-disorder coexisting state appear simultaneously in the Ag12Mo88 multilayered sample and extend over the entire bright field image with unanimously homogeneous composition. In thermodynamic modeling, a Gibbs free energy diagram of the Ag-Mo system is constructed, based on Miedema's model, and suggests that within a narrow composition regime of 85-90 atom % of Mo, the energy difference between the bcc and the amorphous phases is extremely small, which is probably the very reason for the order-disorder coexisting state to appear. In atomistic modeling, an ab initio derived Ag-Mo potential is applied to perform molecular dynamics simulations. The simulations not only determine an intrinsic glass-forming ability/range (GFA/GFR) of the Ag-Mo system to be from 10 to 88 atom % of Mo but also reveal the possibility of the formation/appearance of a crystalline and amorphous mixture in a narrow composition regime of 88-92 atom % of Mo. Apparently, the theoretical results are in excellent agreement and/or compatible with the experimental observations in ion beam mixing. PMID- 16471573 TI - Molecular dynamics simulations of electron-alkali cation pairs in bulk water. AB - The structural, dynamic, and thermodynamic properties of an excess electron interacting with an alkali cation (Na+, K+, Li+) in bulk water were investigated by means of a mixed quantum-classical molecular dynamics simulation technique. This study includes a reparametrization of the electron-cation pseudopotentials. The free energy calculations for all three systems show that a contact electron cation pair can be observed, which is either as stable as the dissociated pair (Li+) or more stable by only a few kT (Na+, K+). Given that the dissociation barrier is also quite small, we suggest that the average cation-electron distance in the experiments at room temperature will not depend on this free energy profile but rather on the minimization of the Coulombic repulsive interaction between like charges in the solvent medium. This enables us to compare the present molecular dynamics simulations with the spectroscopic data obtained for different ionic strengths. The overall trend of the UV-vis hydrated absorption spectra, namely, the shift toward shorter wavelengths at high ionic strengths, is fairly well reproduced. This confirms our hypothesis of statistical distribution of the cations and solvated electrons. PMID- 16471575 TI - The crystal structure and surface energy of NaAlH4: a comparison of DFT methodologies. AB - This paper presents a comparison of the bulk structure, cleavage energies, and local densities of states of solid NaAlH4 calculated using several different density functional theory methodologies. Good agreement is obtained for the bulk crystal structure. Larger differences become apparent for the calculated surface energies and local densities of states. The (001) NaAlH4 surface is clearly identified as the most stable surface, followed by the (112) and (101) surfaces, with the (100) surface being the least stable. We present an analysis of the local density of states of atoms in the exposed NaAlH4 surface. PMID- 16471574 TI - Temperature effects on the structural and dynamical properties of the Zn(II) water complex in aqueous solution: a QM/MM molecular dynamics study. AB - An ab initio quantum mechanical/molecular mechanical (QM/MM) molecular dynamics (MD) simulation at double-zeta restricted Hartree-Fock (RHF) level was performed at an elevated temperature of 363 K (90 degrees C) to study the temperature effects on the structural and dynamical properties of a Zn(II)-water complex in aqueous solution. The first hydration shell, consisting of 6 water molecules at a mean Zn-O distance of 2.16 A, was found to remain stable also at 90 degrees C with respect to exchange processes. The flexible second shell contains, in average, approximately 27 water ligands. To fully characterize the hydration structure, several other parameters such as radial and angular distribution functions (RDF and ADF) and tilt- and theta-angle distributions were evaluated and compared to data obtained at 298 K (25 degrees C). Temperature effects on the dynamics of the Zn(II)-water complex were studied in terms of water reorientations, mean ligand residence times (MRTs), and number of ligand exchange processes. To get further insight into the solute dynamics, additional data, in particular, librational and vibrational motions of water ligands and Zn-O stretching frequencies, were calculated. The second shell is considerably influenced by the elevated temperature, as the ligands' mean residence time is shortened to 4 ps from the value of 10.5 observed at room temperature. The values of the QM/MM MD simulation were also compared to the results of a classical molecular dynamics (CMD) simulation with two- plus three-body potential performed at 90 degrees C, revealing that an accurate description of the second shell and the dynamics of the Zn(II) hydrate needs the inclusion of quantum mechanics in the description. PMID- 16471576 TI - Environmental effect on the relative contribution of the charge-transfer mechanisms within a short DNA sequence. AB - Time evolution of the charge-transfer site population is studied in a short DNA sequence to determine the type of governing charge-transfer mechanism. The system consists of a 5'-GAGGG-3' nucleobase sequence coupled with a dissipative bath that represents the DNA phosphate backbone and solvents. Relative contribution of transfer mechanisms to the whole charge-transfer process has been obtained using the on-the-fly filtered propagator functional path integral method with the density matrix decomposition. Partial density matrixes of the incoherent hopping and coherent superexchange pathways as well as the full reduced density matrix have been evaluated and discussed for both debye and ohmic baths. It was found that the relative contribution of the transfer mechanisms is rather sensitive to the frequency-dependent environmental description. PMID- 16471577 TI - Study of the electrochemical reduction of dioxygen in acetonitrile in the presence of weak acids. AB - The electrochemical reduction of dioxygen has been studied in acetonitrile at glassy-carbon electrodes. The initial step is the reversible one-electron reduction to form superoxide. In the presence of hydrogen-bond donors (water, methanol, 2-propanol), the superoxide forms a complex with the donor resulting in a positive shift in the potential that can be analyzed to obtain formation constants for these complexes. Stronger acids result in protonation of the superoxide followed by reduction to produce HO2-. In the absence of hydrogen-bond donors, the reduction of superoxide occurs at very negative potentials, and this second reduction peak is very much drawn-out along the potential axis, indicating a small value of the transfer coefficient, alpha. The addition of hydrogen-bond donors, HA, brings about a positive shift in this peak, without a noticeable change in shape. The reaction occurring at the second peak is a concerted proton and electron transfer (CPET) in which the electron is transferred to superoxide and a proton is transferred from HA to the superoxide, forming HO2- and A- in a concerted process. An estimation of the standard potential for this reaction shows that the second reduction always occurs at a high driving force, which explains the small value of alpha that is observed. Consistent with a CPET, a kinetic isotope effect, HA versus DA, was detected for the three hydrogen-bond donors. The increasing positive shift of the second peak with increasing water concentration has been interpreted as being a consequence of the change in the formal potential, as water is both a reactant in the process and a participant through the hydrogen-bond stabilization of the anions. PMID- 16471578 TI - Investigation of aerosol formation during benzaldehyde photolysis. AB - The kinetics of photolysis and photonucleation of benzaldehyde (BA) vapor in the air and in an inert gas is investigated, along with the physical and chemical characteristics of the resulting aerosol particles. Short-lived free radicals that accompany the BA photonucleation process were identified by means of spin trapping. Numerical simulation of BA photonucleation, combined with the proposed chemical mechanism, allowed us to calculate the rate of generation of initial particles and to estimate the aerosol quantum yield. PMID- 16471580 TI - Contrasting melting behavior of zinc stearate and zinc oleate. AB - The influence of a double bond in the middle of an otherwise flexible hydrocarbon chain on the melting of such assemblies has been investigated by comparing the melting behavior of zinc stearate and zinc oleate. By monitoring features in the infrared spectra that are characteristic of the global conformation of the hydrocarbon chain, it is shown that the double bond effectively decouples the thermal evolution of conformational disorder in the chain segments on either side of the double bond and the melting of each of these segments in the assembly occurs as independent events. PMID- 16471581 TI - Understanding hydrogen adsorption in metal-organic frameworks with open metal sites: a computational study. AB - Recent experimental investigations show that the open metal sites may have a favorable impact on the hydrogen adsorption capacity of metal-organic frameworks (MOFs); however, no definite evidence has been obtained to date and little is known on the interactions between hydrogen and the pore walls of this kind of MOFs. In this work, a combined grand canonical Monte Carlo simulation and density functional theory calculation is performed on the adsorption of hydrogen in MOF 505, a recently synthesized MOF with open metal sites, to provide insight into molecular-level details of the underlying mechanisms. This work shows that metal oxygen clusters are preferential adsorption sites for hydrogen, and the strongest adsorption of hydrogen is found in the directions of coordinatively unsaturated open metal sites, providing evidence that the open metal sites have a favorable impact on the hydrogen sorption capacity of MOFs. The storage capacity of hydrogen of MOF-505 at room temperature and moderate pressures is predicted to be low, in agreement with the outcome for hydrogen physisorption in other porous materials. PMID- 16471582 TI - Atomic-scale roughness effect on capillary force in atomic force microscopy. AB - We study the capillary force in atomic force microscopy by using Monte Carlo simulations. Adopting a lattice gas model for water, we simulated water menisci that form between a rough silicon-nitride tip and a mica surface. Unlike its macroscopic counterpart, the water meniscus at the nanoscale gives rise to a capillary force that responds sensitively to the tip roughness. With only a slight change in tip shape, the pull-off force significantly changes its qualitative variation with humidity. PMID- 16471583 TI - Pinning mononuclear Au on the surface of titania. AB - We have deposited Au atoms on the surface of titania without sintering or surface damage. Mass-selected Au+ atoms were deposited from the gas phase at room temperature with kinetic energies from <3 to 190+/-3.5 eV. Scanning tunneling microscopy reveals island formation following deposition at <3 eV, while mainly atomic features are observed for energies between approximately 35 and approximately 190 eV. A mixture of islands and atomic features is observed at a landing energy of 20+/-3.5 eV, suggesting a critical energy above which pinning occurs. Cluster size is also probed as a function of coverage in the deposition of Au+ with 100 eV of energy, revealing that sintering begins at a coverage of only 0.06 ML. These observations suggest a mechanism in which high-energy collision leads to the annealing of any impact-created surface damage and the pinning of Au atoms to the surface. We provide a new method of preparing isolated Au atoms on an oxide surface, which can serve as a platform for catalytic studies. PMID- 16471584 TI - Highly optimized fourth-order short-time approximation for path integrals. AB - We derive a fourth-order short-time approximation for use in imaginary-time path integral simulations. The short-time approximation converges for all continuous and bounded-from-below potentials, attains quartic order of convergence for sufficiently smooth potentials, and utilizes statistically independent random variables for its construction. These properties recommend the approximation as a natural replacement of the trapezoidal Trotter-Suzuki approximation for physical systems with continuous distributions. PMID- 16471585 TI - Multigram scale, solventless, and diffusion-controlled route to highly monodisperse PbS nanocrystals. AB - High-quality PbS nanocrystals were produced in multigram-scale quantities through a solventless, heterogeneous, and relatively green route. The heterogeneous nature of this reaction allows one to limit the diffusion in the system, allowing for unprecedented monodispersity and quality of the product demonstrated by a full-width at half-maximum of the photoluminescence peak (PL fwhm) as low as 52 meV, a Stokes shift as low as 10 meV, and a quantum yield (QY) of 40%. The growth of the nanocrystals is interpreted in the framework of a diffusion-controlled Ostwald growth in conditions of strong supersaturation. PMID- 16471586 TI - Ultrafast electron transfer from oligo(p-phenylene-ethynylene)thiol to gold. AB - The electron transfer dynamics of oligo(p-phenylene-ethynylene) (OPE) SAM on Au(111) was studied by resonant photoemission spectroscopy. The ultrafast electron transfer from OPE molecules to Au substrate was clearly observed. The time scale for this charge transfer is much less than 6 fs, the core-hole lifetime for C 1s. This strongly suggests that there is an intense interfacial electronic coupling between OPE molecules and the Au substrate. PMID- 16471587 TI - Chemoselective alkane oxidation by superoxo-vanadium(V) in vanadosilicate molecular sieves. AB - Electron paramagnetic resonance (EPR) spectroscopy of reactive superoxo vanadium(V) species in vanadosilicate molecular sieves (microporous VS-1 and mesoporous V-MCM-41) generated on contact with H2O2, tert-butyl hydroperoxide (TBHP), or (H2+O2) is reported for the first time. By suitable choice of the silicate structure, solvent, and oxidant, we could control the vanadium-(O2-*) bond (i.e., the V-O bond) covalency, the mode of O-O cleavage (in the superoxo species), and, therefore, chemoselectivity in the oxidation of n-hexane: Oxidation by TBHP over V-MCM-41, for example, yielded 27.2% of (n-hexanol+n hexanal+n-hexanoic acid), among the highest chemoselectivities for oxidation of the terminal -CH3 in a linear paraffin reported to date. Over these vanadosilicates, oxidation of the primary C-H bond occurs only via a homolytic O O bond cleavage; the secondary C-H bond oxidations may proceed via both the homo- and heterolytic O-O cleavage mechanisms. PMID- 16471589 TI - Chemisorption-induced spin symmetry breaking in gold clusters and the onset of paramagnetism in capped gold nanoparticles. AB - We present a simple model to describe the induction of magnetic behavior on gold clusters upon chemisorption of one organic molecule with different chemical linkers. In particular, we address the problem of stability of the lowest lying singlet and show that for some linkers there exists a spin symmetry-breaking that lowers the energy and leads to preferential spin density localization on the gold atoms neighboring the chemisorption site. The model is basically an adaptation of the Stoner model for itinerant electron ferromagnetism to finite clusters and it may have important implications for our understanding of surface magnetism in larger nanosystems and its relevance to electronic transport in electrode molecule interfaces. PMID- 16471588 TI - Surface states of titanium dioxide nanoparticles modified with enediol ligands. AB - Control of surface states of titanium dioxide nanoparticles using 2-(3,4 dihydroxyphenyl)ethylamine (dopamine) and 3,4-dihydrophenylacetic acid, which act as ligands to the undercoordinated surface sites (carrier traps), is demonstrated by electrochemical techniques. The deepest traps were found to be most reactive and are selectively removed by the addition of the ligands which enhances the kinetics of electron accumulation in the film. Furthermore, a shift in the Fermi level to more positive potentials was detected for electrodes modified with the negatively charged ligand (3,4-dihydrophenylacetic acid) compared to that of electrodes modified with the positively charged ligand (dopamine). The presence of the negative charge on the ligand also contributed to the underpotential of hydrogen evolution on 3,4-dihydrophenylacetic acid-modified electrodes. PMID- 16471590 TI - Vibrational spectrum of katoite Ca3Al2[(OH)4]3: a periodic ab initio study. AB - The vibrational spectrum of the Si-free katoite hydrogarnet (116 atoms in the unit cell) has been calculated at the periodic ab initio quantum mechanical level with the CRYSTAL program, by using a Gaussian type basis set and the hybrid B3LYP Hamiltonian. The harmonic frequencies at the Gamma point have been obtained by diagonalizing the mass-weighted Hessian matrix, that is evaluated by numerical differentiation of the analytical first derivatives of the energy with respect to the atomic Cartesian coordinates. The parameters controlling the numerical differentiation, as well as the numerical integration of the exchange-correlation functional for the self-consistent field (SCF) calculation, are shown to affect the obtained frequencies by less than 3 cm-1. Before diagonalization, the dynamical matrix is transformed to a block diagonal form according to the irreducible representations of the point group, so that the 345 vibrational modes are automatically classified by symmetry. Various tools are adopted (graphical representation, isotopic substitution, "freezing" part of the unit cell) that permit a complete classification of normal modes and, in particular, an analysis of the modes in terms of simple models (octahedra modes, Ca modes, H stretching, bending, rotations). The harmonic OH stretching band (48 modes) is quite narrow (20 cm-1), indicating that the interaction among OH groups is very weak. As the OH stretching modes are known to be totally separable from the other modes and strongly anharmonic, the one-dimensional Schroedinger equation for the anharmonic oscillator is solved numerically for the two extreme situations, corresponding to the vibration of one decoupled OH and of all 48 OH groups moving in phase. The anharmonic frequencies are 3682 and 3673 cm-1, respectively, in good agreement with IR experiments (a single band at 3661 cm-1 with a width at half band height of 33 cm-1) and confirming that the interaction between OH groups is extremely weak. PMID- 16471591 TI - Synthesis and growth mechanism of titanate and titania one-dimensional nanostructures self-assembled into hollow micrometer-scale spherical aggregates. AB - Three-dimensional, dendritic micrometer-scale spheres of alkali metal hydrogen titanate 1D nanostructures (i.e., nanowires and nanotubes) have been generated using a modified hydrothermal technique in the presence of hydrogen peroxide and an alkali metal hydroxide solution. Sea-urchin-like assemblies of these 1D nanostructures have been transformed into their hydrogen titanate analogues (lepidocrocite HxTi2-x/4squarex/4O4 (x approximately 0.7, square: vacancy)) by neutralization as well as into their corresponding anatase TiO2 nanostructured counterparts through a moderate high-temperature annealing dehydration process without destroying the 3D hierarchical structural motif. The as-prepared hollow spheres of titanate and titania 1D nanostructures have overall diameters, ranging from 0.8 to 1.2 microm, while the interior of these aggregates are vacuous with a diameter range of 100 to 200 nm. The constituent, component titanate and TiO2 1D nanostructures have a diameter range of 7+/-2 nm and lengths of up to several hundred nanometers. A proposed two-stage growth mechanism of these hollow micrometer-scale spheres was supported by time-dependent scanning electron microscopy, atomic force microscopy, and inductively coupled plasma atomic emission spectrometry data. We have also demonstrated that these assemblies are active photocatalysts for the degradation of synthetic Procion Red dye under UV light illumination. PMID- 16471592 TI - Molecular aluminum hydrides identified by inelastic neutron scattering during H2 regeneration of catalyst-doped NaAlH4. AB - Catalyst-doped sodium aluminum hydrides have been intensively studied as solid hydrogen carriers for onboard proton-exchange membrane (PEM) fuel cells. Although the importance of catalyst choice in enhancing kinetics for both hydrogen uptake and release of this hydride material has long been recognized, the nature of the active species and the mechanism of catalytic action are unclear. We have shown by inelastic neutron scattering (INS) spectroscopy that a volatile molecular aluminum hydride is formed during the early stage of H2 regeneration of a depleted, catalyst-doped sodium aluminum hydride. Computational modeling of the INS spectra suggested the formation of AlH3 and oligomers (AlH3)n (Al2H6, Al3H9, and Al4H12 clusters), which are pertinent to the mechanism of hydrogen storage. This paper demonstrates, for the first time, the existence of these volatile species. PMID- 16471593 TI - Unusual coordination behavior of Cr3+ in microporous aluminophosphates. AB - A CrAPO-5 molecular sieve has been investigated with X-ray absorption spectroscopy (EXAFS-XANES) as dehydrated material and after loading with water and ammonia to unravel the coordination geometries of Cr3+ in the framework of a microporous crystalline aluminophosphate, more particularly of the AFI-type. A comparison of the XANES data, a preedge analysis with crystal field multiplet calculations and EXAFS data, pointed toward the presence of framework Cr3+ which, on dehydration, takes on a distorted tetrahedral coordination state. Due to the 3d3 configuration of Cr3+, this unusual tetrahedral coordination environment strongly tends to transform into the more stable 6-fold coordination geometry by binding two extraframework water molecules during hydration. In the presence of ammonia, tetrahedral Cr3+ readily transforms into a 5-fold coordination geometry by binding one ammonia molecule. Therefore, depending on the environmental conditions, the Cr3+ ions can occur in a 4-, 5-, or 6-fold coordination. This observation underlines the flexibility of transition metal ions, such as Cr3+, to cope with unusual coordination geometries in inorganic hosts, making them interesting as potential active sites in heterogeneous catalysis. PMID- 16471594 TI - Solid-state NMR identification and quantification of newly formed aluminosilicate phases in weathered kaolinite systems. AB - The weathering of a specimen kaolinite clay was studied over the course of 369 d via solid-state 29Si magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectroscopy and high-field 27Al MAS NMR. The chosen baseline solution conditions (0.05 mol kg-1 of Al, 2 mol kg-1 of Na+, 1 mol kg-1 of NO3-, 1 mol kg-1 of OH-, and pH approximately 13.8) approximate those of solutions leaking from waste tanks at the Hanford Site in Richland, WA. Nonradioactive Cs and Sr cations were added to this synthetic tank waste leachate (STWL) solution at concentrations of 10(-3), 10(-4), and 10(-5) molal (m) to represent their radionuclide counterparts. The transformations of silicon- and aluminum-containing solid phase species were monitored quantitatively by using NMR spectroscopy, with the resulting spectra directly reporting the influence of the initial Cs and Sr on formation and transformation of the neo-formed solids. At the lowest concentration of Cs and Sr employed (10(-5) m in each cation) peaks consistent with the formation of zeolite-like minerals were detected via 29Si and 27Al MAS NMR as early as 33 d. At concentrations of 10(-3) m in each cation, new silicon species are not detected until 93 d, although neophases containing four coordinate aluminum were detectable at earlier reaction times via 27Al MAS NMR. At the highest magnetic field strengths employed in this NMR study, deconvolutions of resonances detected in the tetrahedral region of the 27Al MAS spectra yielded multiple components, indicating the existence of at least four new aluminum-containing phases. Two of these phases are identified as sodalite and cancrinite through comparison with diffuse-reflectance infrared (DRIFT) spectra and powder X-ray diffraction (XRD) results, while a third phase may correlate with a previously detected aluminum-rich chabazite phase. All measurable solid reaction products have been quantified via their 27Al MAS resonances acquired at high magnetic field strengths (17.6 T), and the quantitative nature of the 27Al NMR data shows that cancrinite growth increases while sodalite reaches a steady state with respect to total aluminum in the solid phases. The data also relate the coupling of Cs sorption to the ripening of feldspathoid phases in this heterogeneous system as a function of time, and illustrate the important influence of co-contaminants on the environmental reaction kinetics studied here. PMID- 16471595 TI - Ultrafast exciton dynamics in CdSe quantum dots studied from bleaching recovery and fluorescence transients. AB - We have performed ultrafast absorption bleach recovery and fluorescence upconversion measurements ( approximately 100 fs time resolution) for three CdSe samples, with nanoparticle diameters of 2.7, 2.9, and 4.3 nm. The two types of experiments provide complementary information regarding the contributions of the different processes involved in the fast relaxation of electrons and holes in the CdSe quantum dots. Transient absorption and emission experiments were conducted for the 1S [1Se-1S3/2(h)] transition, 1S(e) and 1S3/2(h) representing the lowest electron (e) and hole (h) levels. The bleach recovery of the 1S transition shows a approximately 400-500 fs initial rise, which is followed by a size-dependent approximately 10-90 ps decay and finally a long-lived (approximately ns) decay. The fluorescence upconversion signal for the 1S transition shows quite different temporal behavior: a two times slower rise time (approximately 700-1000 fs) and, when the fluorescence upconversion signal has risen to about 20% of its maximum intensity, the signal displays a slight leveling off (bend), followed by a continued rise until the maximum intensity is reached. This bend is well reproducible and power and concentration independent. Simulations show that the bend in the rise is caused by a very fast decay component with a typical time of about 230-430 fs. Considering that the 1S quantum dot excitation is comprised of five exciton substates (F=+/-2, +/-1L, 0L, +/-1U, and 0U), we attribute the disparity in the rise of the bleaching and emission transients to the results from the dynamics of the different excitons involved in respectively the bleaching and fluorescence experiments. More specifically, in transient absorption, population changes of the F=+/-1U excitons are probed, in emission population effects for the F=+/-2 ("dark") and the F=+/-1L ("bright") exciton states are monitored. It is discussed that the fast (approximately 400-500 fs) rise of the bleach recovery is representative of the feeding of the F=+/-1U exciton (by filling of the 1S(e) electron level) and that the slower (approximately 700-1000 fs) feeding of the emissive +/-2, +/-1L excitons is determined by the relaxation of the hole levels within the 1S3/2 fine structure. Finally, the approximately 230-430 fs component, typical of the bend in the fluorescence transient, is attributed to the thermalization of the close-lying +/ 2 ("dark") and +/-1L ("bright") excitons. PMID- 16471596 TI - Epitaxial crystallization of isotactic poly(methyl methacrylate) on highly oriented polyethylene. AB - The annealing behavior of amorphous i-PMMA thin films on highly oriented HDPE substrates was studied by transmission infrared spectroscopy and electron diffraction. The i-PMMA thin film on highly oriented HDPE exhibits a much faster crystallization rate than usual, providing not only a good method for the preparation of crystalline i-PMMA thin and ultrathin film, but also the convenience to observe the crystallization process by infrared spectroscopy in situ. The overall crystallization kinetics of the i-PMMA thin film on the highly oriented HDPE layer was also explored in this work, and an Avrami exponent of about 2 was obtained. The accelerated crystallization behavior indicates a special interaction between HDPE and i-PMMA, which favors the nucleation and crystallization of i-PMMA. This special interaction leads also to an oriented alignment of i-PMMA on the HDPE substrate with both polymer chains parallel, i.e., the occurrence of heteroepitaxy, which could be verified by the polarized infrared spectra and electron diffraction pattern. Electron diffraction analysis further demonstrated that the contact planes of this epitaxial system are (100) lattice planes of both polymers. This can be explained in terms of a two dimensional lattice matching. PMID- 16471597 TI - Solid-state NMR study of Na versus K doping of para-phenylene oligomers. AB - 13C solid-state nuclear magnetic resonance (NMR) experiments were performed on p terphenyl, p-quaterphenyl, and p-sexiphenyl either in their pristine or doped with alkali metals form. The 13C NMR spectra of doped materials show new resonances by comparison with pristine compounds. For the K-doped materials, these resonances appear in the 90-135 ppm range, while for Na-doped materials, they are observed in the larger 20-150 ppm range. It suggests that the interaction between the alkali ions and the oligomers depends on the nature of the alkali. It is corroborated by 13C NMR experiments after exposure to air that show different behaviors. As expected, air exposure of K-doped samples restores the pristine spectra. This is not the case for Na doping, where the signature of the doped material persists even after exposure to air. In the latter case, some 13C resonances can be assigned to sp3 hybridized carbons and to the quinoid group. It suggests that Na doping induces a polymerization of the oligophenylenes. PMID- 16471598 TI - Growth mechanism of penniform BaWO4 nanostructures in catanionic reverse micelles involving polymers. AB - The formation of penniform BaWO4 nanostructures made of nanowires or nanobelts under the direction of a block copolymer in catanionic reverse micelles has been studied in detail. On the basis of the experimental results obtained from the BaWO4 crystallization in aqueous polymer solutions and careful transmission electron microscopy (TEM) observations of BaWO4 nanostructures formed in reverse micelles containing polymers, a detailed two-stage growth mechanism has been proposed for the formation of the penniform nanostructures in reverse micelles, which involves the polymer-controlled shaft formation (Stage 1) and the mixed surfactants-controlled barb growth (Stage 2). During Stage 1, poly(ethylene glycol)-block-poly(methacrylic acid) (PEG-b-PMAA) induced the formation of c-axis oriented shuttle-like nanocrystals and the subsequent oriented attachment of these shuttle-like nanocrystals resulted in the formation of [100]-oriented shafts with many parallel [001]-oriented pricks. During Stage 2, [001]-oriented nanowires or nanobelts grew gradually from the pricks into barbs, leading to the formation of well-defined penniform BaWO4 nanostructures with the barb morphology essentially determined by the mixing ratio r of the anionic to cationic surfactants (i.e., nanowires were formed at r=1 while nanobelts were formed at r deviating from 1). The current understanding of the growth mechanism of penniform BaWO4 nanostructures in catanionic reverse micelles involving polymers may be potentially applied for designing a new synthesis system for the controlled synthesis of other hierarchical 1D nanostructures with desired architectures. PMID- 16471599 TI - Effect of added poly(oxyethylene)dodecyl ether on the phase and rheological behavior of wormlike micelles in aqueous SDS solutions. AB - Upon the addition of a short EO chain nonionic surfactant, poly(oxyethylene) dodecyl ether (C12EOn), to dilute micellar solution of sodium dodecyl sulfate (SDS) above a particular concentration, a sharp increase in viscosity occurs and a highly viscoelastic micellar solution is formed. The oscillatory-shear rheological behavior of the viscoselastic solutions can be described by the Maxwell model at low shear frequency and combined Maxwell-Rouse model at high shear frequency. This property is typical of wormlike micelles entangled to form a transient network. It is found that when C12EO4 in the mixed system is replaced by C12EO3 the micellar growth occurs more effectively. However, with the further decrease in EO chain length, phase separation occurs before a viscoelastic solution is formed. As a result, the maximum zero-shear viscosity is observed at an appropriate mixing fraction of surfactant in the SDS-C12EO3 system. We also investigated the micellar growth in the mixed surfactant systems by means of small-angle X-ray scattering (SAXS). It was found from the SAXS data that the one dimensional growth of micelles was obtained in all the SDS-C12EOn (n=0-4) aqueous solutions. In a short EO chain C12EOn system, the micelles grow faster at a low mixing fraction of nonionic surfactant. PMID- 16471600 TI - Study of molecular dynamics and the solid state phase transition mechanism for unsymmetrical thiopyrophosphate using X-ray diffraction, DFT calculations and NMR spectroscopy. AB - Differential scanning calorimetry (DSC) and low-temperature X-ray diffraction studies showed that 2-thio-(5,5-dimethyl-1,3,2-dioxaphosphorinanyl)2'-oxo dineopentyl-thiophosphate (compound 1) undergoes reversible phase transition at 203 K related to the change of symmetry of the crystallographic unit. Solid state NMR spectroscopy was used to establish the dynamic processes of aliphatic groups and the phosphorus skeleton. 13C and 31P variable temperature NMR studies as well as T1 and T1rho measurements of relaxation times revealed the different mode of molecular motion for each neopentyl residue directly bonded to phosphorus. It is concluded that molecular dynamics of aliphatic groups causes different van der Waals interactions in the crystal lattice and is the driving force of phase transition for compound 1. Finally, we showed that very sharp phase transition temperature makes compound 1 an excellent candidate as a low-temperature NMR thermometer in the solid phase. PMID- 16471601 TI - Layered double hydroxide-CdSe quantum dot composites through colloidal processing: effect of host matrix-nanoparticle interaction on optical behavior. AB - Layered double hydroxide (LDH)-monodispersed 4-nm CdSe nanoparticle composites were prepared through restacking of layers of colloidally dispersed delaminated LDH in the presence of CdSe nanoparticles in 1-butanol. The composites exhibit a blue shift for CdSe absorption, which increases with a decrease in nanoparticle content. The observed blue shift is due to the interaction of the quantum dots with the LDH layers, which leads to surface modification of the nanoparticles. PMID- 16471602 TI - Raman scattering studies of the Ba2MnWO6 and Sr2MnWO6 double perovskites. AB - Polycrystalline Ba2MnWO6 (BMW) and Sr2MnWO6 (SMW) samples were studied between 80 and 1200 K by Raman scattering spectroscopy. In the case of BMW (space group Fmm), four Raman active vibrational modes, predicted by factor group analysis, were identified. Raman scattering studies with different wavelengths revealed a resonant bands between 300 and 800 cm-1. The origin of these bands was related to the Franck-Condon process. Line broadening versus temperature and phonon frequency were studied, and a qualitative explanation was proposed. SMW samples had considerably more complex Raman spectra. It was found that SMW transformed from tetragonal (room-temperature space group P42/n) to the cubic phase between 670 and 690 K; the phase transition temperature was dependent on sample preparation conditions, and it was considerably lower than in the case of large grain size powders. The role of grain size in phase transition is discussed. Mn ions were found to have a crucial role in the lattice dynamics of both materials. PMID- 16471603 TI - Location of phenothiazine in sodium dodecyl sulfate/n-pentanol/water microemulsions. AB - The location of phenothiazine (PTZ) in sodium dodecyl sulfate (SDS)/n-pentanol (n C5H11OH)/water microemulsions is studied by cyclic voltammetry at a Pt electrode. The results indicate that PTZ exists in the membrane phase of microemulsion droplets with its N atom or S atom toward the polar head of the surfactant. In addition, we examine the effect of the compositions and structures of the microemulsions, pH, temperature, and the inorganic salts on the location distribution for PTZ in the membrane phase of the microemulsions. The results show that the location distribution for PTZ in the membrane phase of the microemulsions is mainly dependent on the hydrogen bond between the -NH in PTZ and n-pentanol (or the -SO4- of SDS) and on the electrostatic interaction between the S atom (or N atom) in PTZ and the polar head of SDS. PMID- 16471604 TI - Synthesis of single crystalline tellurium nanotubes with triangular and hexagonal cross sections. AB - Single crystalline tellurium (Te) nanotubes with triangular cross sections were successfully synthesized for the first time by a simple approach of vaporizing tellurium metal and condensing the vapor in an inert atmosphere onto a suitable substrate. Tellurium gas was evaporated by heating at 350 degrees C and was condensed on the Si (100) substrate at 150-200 degrees C, in the downstream of argon (Ar) gas at a flow rate of 25 sccm for 10 min. This led to the production of nanotubes of triangular cross section along with some hexagonal ones. The formation of the nanotubes was highly dependent upon the structure of the substrate surface, Ar gas flow rate, and the deposition temperature. When the substrate is Si (111) or sapphire (0001) or when the argon flow rate is increased to 500 sccm, nanowires and nanorods were exclusively formed. Irrespective of the morphologies, all the observed Te nanostructures grew in a regular [0001] direction. The facile approach to nanotubes with a triangular cross section may facilitate some new applications as well as stimulate theoretical studies pertaining to the stability of this high-energy configuration. PMID- 16471605 TI - Preparation and electrical properties of ultrafine Ga2O3 nanowires. AB - Uniform and well-crystallized beta-Ga2O3 nanowires are prepared by reacting metal Ga with water vapor based on the vapor-liquid-solid (VLS) mechanism. Electron microscopy studies show that the nanowires have diameters ranging from 10 to 40 nm and lengths up to tens of micrometers. The contact properties of individual Ga2O3 nanowires with Pt or Au/Ti electrodes are studied, respectively, finding that Pt can form Schottky-barrier junctions and Au/Ti is advantageous to fabricate ohmic contacts with individual Ga2O3 nanowires. In ambient air, the conductivity of the Ga2O3 nanowires is about 1 (Omega.m)-1, while with adsorption of NH3 (or NO2) molecules, the conductivity can increase (or decrease) dramatically at room temperature. The as-grown Ga2O3 nanowires have the properties of an n-type semiconductor. PMID- 16471606 TI - Preparation and catalytic performances of ultralarge-pore TiSBA-15 mesoporous molecular sieves with very high Ti content. AB - Highly ordered TiSBA-15 mesoporous molecular sieves with different nSi/nTi ratios and tunable pore diameters have been prepared through direct synthesis under various hydrochloric acid concentrations and synthetic temperatures. The structure and the textural parameters of the materials were investigated by powder X-ray diffraction and nitrogen adsorption/desorption measurements. Decrease of the acid concentration and nSi/nTi ratio in the synthetic gel enhanced the amount of Ti incorporation in SBA-15 materials without affecting their structural order and textural parameters. Highly ordered mesoporous TiSBA 15 with a very high Ti content up to a nSi/nTi ratio of 1.9 was prepared for the first time under the optimized synthesis conditions. Control of synthetic temperature resulted in tuning of pore geometries without structural deterioration and Ti content. Ultralarge-pore TiSBA-15 with a pore size of 12.6 nm and a pore volume of 1.3 cm3 g-1 was also synthesized. The nature and the coordination of the Ti atoms in SBA-15 prepared under various synthesis conditions were investigated by UV-vis spectroscopy. It has been found that the Ti atoms are well-dispersed and mostly occupy the tetrahedral coordination under the optimized synthesis conditions. Catalytic performance of the obtained TiSBA 15 materials was also investigated through oxidation of styrene by hydrogen peroxide and tert-butylhydroperoxide as oxidants. PMID- 16471607 TI - A vapor-solid strategy to silica sheathed metal nanostructures and microstructures via reactions of metal chlorides with silicon. AB - A facile vapor-solid strategy has been developed to prepare silica-sheathed metal micro/nanostructures with controllable shapes. As examples, silica-sheathed nickel nanowires (diameter approximately 50 nm), microcubes (edge length 1-3 microm), nanocubes (edge length approximately 200 nm) with an epitaxial tail (diameter<100 nm), and 1D assembly structures of nanoparticles (particle diameter<100 nm) as well as silica-sheathed cobalt and copper micro/nanostructures are synthesized. The possible reaction and growth mechanisms of Ni/SiO2 structures are discussed. The method is expected to be applied to a wider range of metals. PMID- 16471608 TI - First evidence of simultaneous different kinetic behaviors at the interface and the continuous medium of w/o microemulsions. AB - A study was carried out on the butylaminolysis reaction of 4-nitrophenyl caprate in AOT/chlorobenzene/water microemulsions, with the observed rate constant, kobs, showing both first- and second-order dependence on butylamine concentration. The first-order term in [BuNH2] is due to the reaction occurring at the interface of the microemulsion while the second-order term is due to the reaction in the continuous medium. The different kinetic behavior is accounted for by the mechanism by which the reaction proceeds: at the interface of the microemulsion, the rate-determining step is the formation of the addition intermediate, T+/-, whereas in the continuous medium the slow step is the base-catalyzed decomposition of this intermediate. The application of the pseudophase formalism allows the observed kinetic behavior to be explained and to obtain the rate constants at the interface, ki2=0.13 M-1 s-1, and in the continuous medium, ko2KT=2.46x10(-2) M-2 s-1. These values indicate that the reaction rate decreases approximately 23 times upon going from the aqueous medium to the interface of the microemulsion, whereas the rate constant in the continuous medium is consistent with that obtained in pure chlorobenzene, ko2KT=2.09x10(-2) M-2 s-1. PMID- 16471609 TI - High density germanium nanowire assemblies: contact challenges and electrical characterization. AB - The conductive properties of vertically aligned germanium nanowires, with mean diameters of 50 and 100 nm, within anodized aluminum oxide (AAO) templates have been characterized by conductive atomic force microscopy (C-AFM) and macrocontact measurements. C-AFM was used to determine the electrical transport properties of individual nanowires within the arrays, while macrocontacts were used to measure the mean current-voltage characteristics of groups of nanowires. Contact resistance between the nanowires and metal macrocontacts was minimized by polishing and gradual etching of the AAO surface, to expose the nanowires, prior to deposition of the contacts. Impedance measurements were used to analyze the importance of defects on the charge transport properties of the germanium nanowire arrays. Conductivity data from C-AFM and macrocontact measurements were found to be comparable suggesting that both methods are inherently suitable for evaluating the electrical transport properties of encapsulated nanowires within a matrix. These results are significant as the ability to make good ohmic contacts to nanowires, within well-defined arrays, is key for the future "bottom-up" fabrication of multilayered device architectures for future electronic and optoelectronic devices. PMID- 16471610 TI - Straight and thin ZnO nanorods: hectogram-scale synthesis at low temperature and cathodoluminescence. AB - A novel seed-assisted chemical reaction at 95 degrees C has been employed to synthesize uniform, straight, thin, and single-crystalline ZnO nanorods on a hectogram scale. The molar ratio of ZnO seed and zinc source plays a critical role in the preparation of thin ZnO nanorods. At a low molar ratio of ZnO seed and zinc source, javelin-like ZnO nanorods consisting of thin ZnO nanorods with a diameter of 100 nm and thick ZnO nanorods with a diameter of 200 nm have been obtained. In contrast, straight ZnO nanorods with a diameter of about 20 nm have been prepared. Dispersants such as poly(vinyl alcohol) act spatial obstructors to control the length of ZnO nanorods. The morphology, structure, and optical property of the ZnO nanostructures prepared under different conditions have been characterized by transmission electron microscopy, field emission scanning electron microscopy, X-ray powder diffraction, high-resolution transmission electron microscopy, and cathodoluminescence. The formation mechanisms for the synthesized nanostructures with different morphologies have been phenomenologically presented. PMID- 16471611 TI - Isolation and characterization of fluorescent nanoparticles from pristine and oxidized electric arc-produced single-walled carbon nanotubes. AB - Fluorescent nanoparticles were isolated from both pristine and nitric acid oxidized commercially available carbon nanotubes that had been produced by an electric arc method. The pristine and oxidized carbon nanotube-derived fluorescent nanoparticles exhibited a molecular-weight-dependent photoluminescence in the violet-blue and blue to yellowish-green ranges, respectively. The molecular weight dependency of the photoluminescence was strongly related to the specific supplier. We analyzed the composition and morphology of the fluorescent nanoparticles derived from pristine and oxidized nanotubes from one supplier. We found that the isolated fluorescent materials were mainly composed of calcium and zinc. Moreover, the pristine carbon nanotube derived fluorescent nanoparticles were hydrophobic and had a narrow distribution of maximal lateral dimension. In contrast, the oxidized carbon nanotube-derived fluorescent nanoparticles were superficially oxidized and/or coated by a thin carbon layer, had the ability to aggregate when dispersed in water, and exhibited a broader distribution of maximal lateral dimension. PMID- 16471612 TI - Study on the synthesis of poly(diglycidyl maleate-co-stearyl methacrylate) and morphology conversion of their self-assembly systems. AB - A novel polymer of poly(diglycidyl maleate-co-stearyl methacrylate) (P(DGMA-co SMA)) was synthesized by reaction between poly(maleic anhydride-co-stearyl methacrylate) (P(MA-co-SMA)) and epichlorohydrin. The self-assembly behavior of the resultant copolymer was investigated. It was found that the spheral aggregates could converse to nanorods after being aged for 2.5 days and nanolines composed of the nanorods were obtained after being aged for an additional 5.5 days. The mechanism of their self-assembly behavior and morphology conversion of self-assembly systems is discussed. PMID- 16471613 TI - Metallic ReO3 nanoparticles. AB - ReO3 nanoparticles in the diameter range of 8.5-32.5 nm have been prepared by decomposition of the Re2O7-dioxane complex under solvothermal conditions and characterized by X-ray diffraction, electron microscopy, optical spectroscopy, and scanning probe microscopy. The nanoparticles have the cubic (Pm3m {221} space group) structure with the lattice parameter increasing with decreasing size. The particles are metallic and show a plasmon band around 520 nm, which becomes blue shifted with a decrease in size. The metallicity of the nanoparticles is also confirmed by tunneling conductance measurements. The nanoparticles show paramagnetic or diamagnetic behavior depending on the size, with evidence for superparamagnetism at low temperatures when the size is small. PMID- 16471614 TI - Photoluminescence investigation based on laser heating effect in ZnO-ordered nanostructures. AB - We synthesized ZnO-SiO2 composite opal and ZnO inverse opal by electrodeposition using SiO2-opal template and polystyrene (PS)-opal template, respectively. Compared with compact ZnO nanocrystal film also prepared by electrodeposition, ordered ZnO nanostructures exhibit more significant red-shift and broadening of the UV peak with increasing excitation power, which is due to a stronger local heating effect in ordered ZnO nanostructures. We developed a quantitative analytical method to investigate photoluminescence (PL) of ZnO based on laser heating effects. The experimental data agree well with fitting curves derived from the electron-phonon interaction model. Important parameters, such as electron-phonon coupling strength and thermal activation energy, can be obtained by fitting experimental data. The resonant Raman spectra provide further evidence that the analyses based on laser heating effects are feasible. PMID- 16471615 TI - In situ synthesis and characterization of multiwalled carbon nanotube/Au nanoparticle composite materials. AB - An effective and facile in situ reduction approach for the fabrication of carbon nanotube-supported Au nanoparticle (CNT/Au NP) composite nanomaterials is demonstrated in this article. Linear polyethyleneimine (PEI) is ingeniously used as both a functionalizing agent for the multiwalled carbon nanotubes (MWNTs) and a reducing agent for the formation of Au NPs. This method involves a simple mixing process followed by a mild heating process. This approach does not need the exhaustive surface oxidation process of CNTs. The coverage of Au NPs on CNTs is tunable by varying the experimental parameters, such as the initial molar ratio of PEI to HAuCl4, the relative concentration of PEI and HAuCl4 to MWNTs, and the temperature and duration of the heat treatment. More importantly, even the heterogeneous CNT/Au composite nanowires are obtainable through this method. TEM, XPS, and XRD are all used to characterize the CNT/Au composite materials. In addition, the optical and electrocatalytic properties are investigated. PMID- 16471616 TI - Calorimetric and FTIR study of the acid properties of sulfated titanias. AB - Titanium oxides of different surface areas were sulfated then calcined to convert the solid to a strong acid. The amount of sulfur retained by the solid and the thermal stability of the resulting sulfate are controlled by the dispersion of the initial oxide. The acid properties were determined by gravimetry at 383 K, calorimetry using ammonia adsorption at 353 K, and by quantitative analysis of the infrared spectra of pyridine retained after evacuation at 423 K. A good agreement was observed between the different determinations. At low coverage of ammonia, sulfated titanias show a much lower heat of adsorption, and the IR study of NH3 adsorption shows that the first doses of NH3 dissociate at the surface with the formation of OH species. The lower heat of adsorption is then attributed to the contribution of NH3 dissociation to the differential heat of adsorption. IR spectroscopy indicates that NH3 reacts with sulfates and may lead to the transformation of disulfate species into monosulfate species on sulfated titania dioxide. A band at ca. 3574 cm-1 has been assigned to nu(OH) of monosulfate species. This particular behavior makes it difficult to appreciate the initial acidity of these sulfated oxides. PMID- 16471617 TI - Presence or absence of counterion specificity in the interaction of alkylammonium surfactants with alkylacrylamide gels. AB - Patterns in the interaction of cationic surfactants with nonionic polymer gels, which were inferred from a recent study from our laboratory, are confirmed by measurements of a series of alkylammonium surfactants with different counterions with a series of alkyl acrylamide gels of increasing hydrophobicity. Two swelling patterns were observed: Either the swelling continued above the surfactant critical micelle concentration (cmc) and the maximum swelling differed for different counterions and increased in the order of Br-15 eV), with no measurable difference between them (i.e., no differential charging between the silicon nanoclusters and the oxide matrix could be detected). By use of a measured Auger parameter, we estimate the relaxation energy of the Si(nc) in the SiO2 matrix as -0.4 eV, which yields a -0.6 eV shift in the binding energy of the Si(nc) with respect to that of bulk Si in the opposite direction of the expected quantum size effect. This must be related to the residual differential charging between the silicon nanoclusters and the oxide host. Therefore, differential charging is still the biggest obstacle for extracting size-dependent binding energy shifts with XPS when one uses the oxide peak as the reference. PMID- 16471656 TI - A combined experimental and theoretical study on the effect of doping and interface formation on Ppv-ether copolymer. AB - We present a detailed optoelectronic and vibrational study devoted to the transformation from neutral to doped PPV-ether copolymer in both powder and thin film states. The full geometries were optimized with the density functional theory (DFT) for neutral and doped states, where a comparative geometric study was established. The lowest singlet excited-state geometries have been investigated by using the configuration interaction single (CIS/3-21G(d)) method. The absorption spectra are then calculated respectively on the basis of the ground- and excited-state geometries. Our calculation results are in close agreement with those available from experiments. The charge distribution and excitation energies of singly charged PPV-ether are calculated, where two subgap absorption features are found to dominate the optical spectrum correlated with the polaron picture. These theoretical results are compared to experimental optical data illustrated by iodine-doped PPV-ether. Next, we have performed a simulation to model the conformations and the electronic structure modifications of interface formation of PPV-ether copolymer thin film with calcium (Ca), magnesium (Mg), and aluminum (Al) metal as a cathode and indium tin oxide (ITO) as anode in polymer LEDs. By providing the optical parameter obtained and the chemical reaction at the interface, we present the energetic diagram near the interface and the energy position of the lowest occupied molecular orbital with respect to the electrode Fermi level. PMID- 16471657 TI - Thermotropic phase behavior of cationic gemini surfactants and their equicharge mixtures with sodium dodecyl sulfate. AB - The lyotropic phase behavior for the neat cationic gemini surfactants alkanediyl alpha,omega-bis(alkyldimethylammonium bromide), designated here as m-s-m, has been investigated previously in several works, but the thermotropic behavior has not been well characterized. Only for 15-s-15 and 14-s-12 have thermotropic liquid crystals (Lc) been reported. In this work, for the first time and in contrast to previous reports, we observe thermotropic Lc formation for m-2-m geminis with m = 12, 14, 16, and 18, by means of polarizing microscopy and differential scanning calorimetry (DSC). Furthermore, we investigate mixtures of m-2-m and SDS, m-2-m Br2.2SDS, which exhibit crystal-to-crystal phase transitions at lower temperature and, at high temperature, smectic Lc phases. The transition temperatures and enthalpies for Lc phases, obtained by DSC, present clear trends upon increase of the chain lengths. Combining Langmuir film experiments, possible lamellar arrangements for the different phases are tentatively discussed. PMID- 16471658 TI - Controllable synthesis of conducting polypyrrole nanostructures. AB - Wire-, ribbon-, and sphere-like nanostructures of polypyrrole have been synthesized by solution chemistry methods in the presence of various surfactants (anionic, cationic, or nonionic surfactant) with various oxidizing agents [ammonium persulfate (APS) or ferric chloride (FeCl3), respectively]. The surfactants and oxidizing agents used in this study have played a key role in tailoring the nanostructures of polypyrrole during the polymerization. It is inferred that the lamellar structures of a mesophase are formed by self-assembly between the cations of a long chain cationic surfactant [cetyltrimethylammonium bromide (CTAB) or dodeyltrimethylammonium bromide (DTAB)] and anions of oxidizing agent APS. These layered mesostructures are presumed to act as templates for the formation of wire- and ribbon-like polypyrrole nanostructures. In contrast, if a short chain cationic surfactant octyltrimethylammonium bromide (OTAB) or nonionic surfactant poly(ethylene glycol) mono-p-nonylphenyl ether (Opi-10) is used, sphere-like polypyrrole nanostructures are obtained, whichever of the oxidizing agents mentioned above is used. In this case, micelles resulting from self assembly among surfactant molecules are envisaged to serve as the templates while the polymerization happens. It is also noted that, if anionic surfactant sodium dodeyl surfate (SDS) is used, no characteristic nanostructures of polypyrrole were observed. This may be attributed to the doping effect of anionic surfactants into the resulting polypyrrole chains, and as a result, micelles self-assembled among surfactant molecules are broken down during the polymerization. The effects of monomer concentration, surfactant concentration, and surfactant chain length on the morphologies of the resulting polypyrrole have been investigated in detail. The molecular structures, composition, and electrical properties of the nanostructured polypyrrole have also been investigated in this study. PMID- 16471659 TI - The electronic structure of the hydrated proton: a comparative X-ray absorption study of aqueous HCl and NaCl solutions. AB - The oxygen K edge X-ray absorption spectra of aqueous HCl and NaCl solutions reveal distinct perturbations of the local water molecules by the respective solutes. While the addition of NaCl leads to large spectral changes, the effect of HCl on the observed X-ray absorption spectrum is surprisingly small. Density functional theory calculations suggest that this difference primarily reflects a strong blue shift of the hydrated proton (in either the Eigen (H9O4+) or Zundel (H2O5+) forms) spectrum relative to that of H2O, indicating the tighter binding of electrons in H3O+. This spectral shift counteracts the spectral changes that arise from direct electrostatic perturbation of water molecules in the first solvation shell of Cl-. Consequently, the observed spectral changes effected by HCl addition are minimal compared to those engendered by NaCl. Additionally, these results indicate that the effect of monovalent cations on the nature of the unoccupied orbitals of water molecules in the first solvation shell is negligible, in contrast to the large effects of monovalent anions. PMID- 16471660 TI - Effect of double bonds on the conducting properties of ciguatoxin 3C and tetrahydropyrane-based polymers: a theoretical study. AB - The electronic structure of the ciguatoxin 3C is analyzed through the Kohn-Sham model by using two different kinds of basis sets: localized basis set (Gaussian functions) and nonlocalized basis set (plane wave functions). With the localized basis functions, two approximations are used for the exchange-correlation functional: the local density approximation and the generalized gradient approximation. With the nonlocalized basis set, just the local density approximation is used. The energy gap, obtained from the frontier molecular orbitals, for this molecule predicts that this system is a semiconductor, even when the number of double bonds is increased inside the structure. However, as large molecules built with the basic unit--the tetrahydropyrane--of the ciguatoxin 3C are found in nature, it suggests studying the gap in polymeric systems built with the basic unit of this molecule. It is demonstrated that the presence of double bonds reduces considerably the gap, indicating the possibility of forming conducting materials by introducing double bonds in this kind of molecular systems. Thus, molecules strongly linked with biological systems can be used as precursor to build electric conducting systems. PMID- 16471661 TI - Thermogravimetric analysis of synthesis variation effects on CVD generated multiwalled carbon nanotubes. AB - Changes in the thermogravimetrically determined oxidation behaviors of CVD-grown multiwalled carbon nanotubes with varying synthesis conditions are examined. Catalyst type and synthesis temperature are found to have a measurable impact upon nanotube stability, suggesting differing levels of crystalline perfection in the resulting nanotubes. The results provide evidence showing the catalytic effects of nanotube catalyst particles and their oxides upon the oxidation of nanotube carbon and graphite. The significance of thermogravimetric analysis as a characterization tool for carbon nanotubes is discussed. PMID- 16471662 TI - Comparison of the magnetic properties of metastable hexagonal close-packed Ni nanoparticles with those of the stable face-centered cubic Ni nanoparticles. AB - We report the first magnetic study of pure and metastable hexagonal close-packed (hcp) Ni nanoparticles (sample 1). We also produced stable face-centered cubic (fcc) Ni nanoparticles, as mixtures with the hcp Ni nanoparticles (samples 2 and 3). We compared the magnetic properties of the hcp Ni nanoparticles with those of the fcc Ni nanoparticles by observing the evolution of magnetic properties from those of the hcp Ni nanoparticles to those of the fcc Ni nanoparticles as the number of fcc Ni nanoparticles increased from sample 1 to sample 3. The blocking temperature (T(B)) of the hcp Ni nanoparticles is approximately 12 K for particle diameters ranging between 8.5 and 18 nm, whereas those of the fcc Ni nanoparticles are 250 and 270 K for average particle diameters of 18 and 26 nm, respectively. The hcp Ni nanoparticles seem to be antiferromagnetic for T < T(B) and paramagnetic for T > T(B). This is very different from the fcc Ni nanoparticles, which are ferromagnetic for T < T(B) and superparamagnetic for T > T(B). This unusual magnetic state of the metastable hcp Ni nanoparticles is likely related to their increased bond distance (2.665 angstroms), compared to that (2.499 angstroms) of the stable fcc Ni nanoparticles. PMID- 16471663 TI - Evolution of the local structure around Ti atoms in NaAlH4 doped with TiCl3 or Ti13.6THF by ball milling using X-ray absorption and X-ray photoelectron spectroscopy. AB - X-ray absorption and X-ray photoelectron spectroscopy are used to investigate NaAlH4 doped with 5 mol % of Ti on the basis of either TiCl3 or Ti13.6THF by ball milling. X-ray photoelectron spectroscopy (XPS) analysis of TiCl3 or Ti colloid doped samples indicates that Ti species do not remain on the sample surface but are driven into the material with increasing milling time. The surface concentration of Ti continues to decrease during subsequent cycles under hydrogen. After several cycles, it reaches a constant value of 0.5 at. % independently of the nature of the precursor. Moreover, metallic aluminum is already present at the surface after 2 min of ball milling in the case of TiCl3 doped Na-alanate, whereas it is totally absent in the case of Ti colloid doped samples at any milling time. Upon cycling, the atomic concentration of metallic Al at the surface evolves with the reaction under hydrogen, in contrast to the Ti concentration. Analysis of the binding energies of samples doped with TiCl3 or Ti colloid, after eight desorption/absorption cycles, reveals that the Na, O, and Ti environment remains the same, while the Al environment undergoes changes. According to the extended X-ray absorption fine structure (EXAFS) analysis of TiCl3 doped Na-alanate, the local structure around Ti during the first cycle is close to that of metallic Ti but in a more distorted state. In the case of the Ti colloid doped sample, a stripping of the oxygen shell occurs. After eight cycles, a similar intermetallic phase between Ti and Al is present in the hydrogenated state of TiCl3 or Ti colloid doped samples. The local structure around Ti atoms after eight cycles consists of Al and Ti backscatterers with a Ti-Al distance of 2.79 angstroms and a Ti-Ti distance of 3.88 angstroms. This local structure is not exactly the TiAl3 phase because it differs significantly from the alloy phase in its fine structure and lacks long-range order. Volumetric measurements performed on these samples indicate that the formation of this local structure is responsible for the reduction of the reversible hydrogen capacity with the increasing number of cycles. Moreover, the formation of the alloy-like phase is correlated with a decrease of the desorption/absorption reaction rate. PMID- 16471664 TI - Improvement of Fe/MgO catalysts by calcination for the growth of single- and double-walled carbon nanotubes. AB - Calcination at 900-1000 degrees C for 8-12 h of an Fe/MgO catalyst prepared by impregnation was found to result in a uniform MgFe2O4/MgO solid solution that showed a successful settling of well-dispersed iron species into the MgO lattice. During methane reduction, many iron-containing particles with a diameter of about 4 nm were formed on the catalyst surface to provide numerous active sites for the growth of single- and double-walled carbon nanotubes. There was a significant improvement of the Fe/MgO catalyst that resulted in a high yield of impurity-free nanotubes. Using C2H4 cracking at 600 degrees C and transmission electron microscope observations, the Fe species distribution in the catalysts and microscope images of nanotube growth were described in detail. H2 reduction of the calcined Fe/MgO catalyst was found to cause the formation of iron layers on the catalyst surface, which resulted in the growth of only carbon layers. The results are useful for understanding changes in the metal species distribution in the catalysts and the nanotube growth mechanism, and they provide a simple method to improve Fe/MgO catalysts. PMID- 16471665 TI - Temperature dependence of the Raman spectra of individual carbon nanotubes. AB - Resonant Raman scattering (RRS) spectra of individual carbon nanotubes on a SiO2 substrate have been investigated first in the temperature range of 100-600 K (Phys. Rev. B 2002, 66, 115411). It was revealed by the intensity abnormality of the radial breathing mode (RBM) that the carbon nanotubes have a temperature dependent density of electronic states. This means that the previously reported temperature coefficients of RBM of carbon nanotubes are smaller than their "real" ones for the bulk samples of single- or double-walled carbon nanotubes. Comparatively, the G line of individual nanotubes shows no observable difference relative to the bulk samples. PMID- 16471666 TI - Adsorption and solar light decomposition of acetone on anatase TiO2 and niobium doped TiO2 thin films. AB - Adsorption and solar light decomposition of acetone was studied on nanostructured anatase TiO2 and Nb-doped TiO2 films made by sol-gel methods (10 and 20 mol % NbO2.5). A detailed characterization of the film materials show that films contain only nanoparticles with the anatase modification with pentavalent Nb oxide dissolved into the anatase structure, which is interpreted as formation of substituted Nb=O clusters in the anatase lattice. The Nb-doped films displayed a slight yellow color and an enhanced the visible light absorption with a red-shift of the optical absorption edge from 394 nm for the pure TiO2 film to 411 nm for 20 mol % NbO2.5. In-situ Fourier transform infrared (FTIR) transmission spectroscopy shows that acetone adsorbs associatively with eta1-coordination to the surface cations on all films. On Nb-doped TiO2 films, the carbonyl bonding to the surface is stabilized, which is evidenced by a lowering of the nu(C=O) frequency by about 20 cm(-1) to 1672 cm(-1). Upon solar light illumination acetone is readily decomposed on TiO2, and stable surface coordinated intermediates are formed. The decomposition rate is an order of magnitude smaller on the Nb-doped films despite an enhanced visible light absorption in these materials. The quantum yield is determined to be 0.053, 0.004 and 0.002 for the pure, 10% Nb:TiO2, and 20%Nb:TiO2, respectively. Using an interplay between FTIR and DFT calculations we show that the key surface intermediates are bidentate bridged formate and carbonate, and H-bonded bicarbonate, respectively, whose concentration on the surface can be correlated with their heats of formation and bond strength to coordinatively unsaturated surface Ti and Nb atoms at the surface. The oxidation rate of these intermediates is substantially slower than the initial acetone decomposition rate, and limits the total oxidation rate at t>7 min on TiO2, while no decrease of the rate is observed on the Nb-doped films. The rate of degradation of key surface intermediates is different on pure TiO2 and Nb-doped TiO2, but cannot explain the overall lower total oxidation rate for the Nb-doped films. Instead the inferior photocatalytic activity in Nb-doped TiO2 is attributed to an enhanced electron-hole pair recombination rate due to Nb=O cluster and cation vacancy formation. PMID- 16471667 TI - Interlayer self-diffusion and structure of chiral smectic phases studied by 2H NMR exchange experiment. AB - A deuterium two-dimensional exchange NMR technique is used to study the chiral smectic C (SmC*) and chiral subphases (SmC*(Fi1), SmC*(Fi2)) of a smectogen (s)-1 methylheptyl 4'-(4-n-decyloxy-benzoyloxy)biphenyl-4-carboxylate (10B1M7). The aim is to demonstrate how this technique can be combined with sample rotation in the magnetic field to obtain the fast translational self-diffusion constant along the helical pitch in the SmC* phase and to shed light on the structure of helicoidal superlattice in the three-layer SmC*(Fi1) and four-layer SmC*(Fi2) phases. The cross-peak intensities in the 2D exchange spectrum are sensitive to the disposition of molecules in the three- and four-layer base unit. The results support the "asymmetric Clock model" as an appropriate description of the ferrielectric phases in this compound. PMID- 16471668 TI - Temperature-dependent Raman scattering of silicon nanowires. AB - Silicon nanowires with narrowly distributed diameters of 20-30 nm have been fabricated by chemical vapor deposition on an anodized aluminum oxide (AAO) substrate. The first-order and second-order Raman scatterings of the silicon nanowires have been studied in a temperature range from 123 to 633 K. Both of the first-order and second-order Raman peaks were found to shift and broaden with increasing temperature. The experimental results were analyzed by combining the phonon confinement effect, anharmonic phonon processes and lattice stress effect. It was found that the intensities of the first-order and second-order Raman bands have different dependences on temperature. The value of relative intensities I(2TA)int/I(2TO)int for silicon nanowires was found to be larger than that of bulk silicon, and increase with rising measurement temperature. We ascribe this phenomenon to the participation of phonons with a large wave vector value of Raman scattering caused by both the phonon confinement effect and the temperature effect. PMID- 16471669 TI - Templated deposition of MoS2 nanotubules using single source precursor and studies of their optical limiting properties. AB - We demonstrate a simple way of fabricating bulk quantities of molybdenum disulfide (MoS2) porous nanotubules (NTs) by templated deposition from a single source precursor based on tetrakis(diethylaminodithiocarbomato)molybdate(IV). Bulk quantities of crystalline MoS2 NTs that consist of agglomerates of nested nanocapsules were obtained by this one-step evaporation method. We found that MoS2 NTs show good optical limiting behavior with 532-and 1064-nm nanosecond laser pulses, and the size-effect of the NTs on optical transmission was observed. PMID- 16471670 TI - Sulfated mesoporous alumina: a highly effective solid strong base catalyst for the Tishchenko reaction in supercritical carbon dioxide. AB - Heterogeneous strong base catalysis for the intramolecular Tishchenko reaction of aromatic 1,2-dicarbaldehydes to the corresponding phthalides in supercritical CO2CscCO2 has been realized with mesoporous alumina containing SO4(2-) ions in the alumina framework (mesoAl2O3/SO4(2-)). Infrared spectroscopy of pyrrole adsorbed on the alumina and strong poisoning by a weak Bronsted acid of methanol revealed that the SO4(2-) ions in the framework slightly suppressed the average strength of base sites (O2-) on mesoAl2O3/SO4(2-), but there exists a small number of strong base sites that promote the Tishchenko reaction in scCO2. Although the intramolecular Tishchenko reaction of phthalaldehyde to phthalide in scCO2 was somewhat slower than those in organic solvents such as tetrahydrofuran (THF) and benzene, the addition of a small amount of THF as a cosolvent remarkably increased the reaction rate; the reaction in the scCO2-THF system proceeded 1.5-fold faster than those in pure benzene and THF solvents. PMID- 16471671 TI - New insights into the interaction of hydrogen atoms with boron-substituted carbon. AB - Boron substitution in carbon materials has been comprehensively investigated using the density functional theory method. It was found that there is a correlation between the stability of the graphene sheet, the distribution of pi electrons, the electrostatic potential, and the capability for hydrogen-atom adsorption. Boron substitution destabilizes the graphene structure, increases the density of the electron wave around the substitutional boron atoms, and lowers the electrostatic potential, thus improving the hydrogen adsorption energy on carbon. However, this improvement is only ca. 10-20% instead of a factor of 4 or 5. Our calculations also show that two substitutional boron atoms provide consistent and reliable results, but one substitutional boron results in contradictory conclusions. This is a warning to other computational chemists who work on boron substitution that the conclusion from one substitutional boron might not be reliable. PMID- 16471672 TI - Charge-transfer effect at the interface of phthalocyanine-electrode contact studied by scanning tunneling spectroscopy. AB - Scanning tunneling microscopy (STM) and spectroscopy (STS) are used in this work to investigate the charge-transfer effect at the molecule-substrate interface of substituted metal phthalocyanines. STS results revealed that the apparent energy gaps for both fluorinated phthalocyanines and unsubstituted phthalocyanines are essentially the same, which agree with the hybrid density functional calculations. More interestingly, there is a systematic shift of the energy level of valence bands, possibly as the result of charge-transfer effect at the molecule-substrate interface. PMID- 16471673 TI - CO adsorption on CoMo and NiMo sulfide catalysts: a combined IR and DFT study. AB - Experimental IR spectra of carbon monoxide adsorbed on a series of Mo/Al2O3, CoMo/Al2O3, and NiMo/Al2O3 sulfided catalysts have been compared to ab initio DFT calculations of CO adsorption on CoMo and NiMo model surfaces. This approach allows the main IR features of CO adsorbed on the sulfide phase to be assigned with an uncertainty of 15 cm(-1). On the CoMo system, the band at 2070 cm(-1) is specific of the promotion by Co and is assigned to CO interacting either with a Co atom or with a Mo atom adjacent to a Co atom. On the NiMo system, CO adsorption on Ni centers of the promoted phase leads to a high-wavenumber band at approximately 2120 cm(-1) that strongly overlaps the band at 2110 cm(-1) characteristic of nonpromoted Mo sites. For NiMo and CoMo catalysts, broad shoulders at low wave numbers (below 2060 cm(-1)) are characteristic of Mo centers adjacent to promoter atoms, indicating a partial decoration of the MoS2 edges by the promoter. PMID- 16471674 TI - Morphology selected molecular architecture: acridine carboxylic acid monolayers on Ag (111). AB - The molecular architecture of acridine-9-carboxylic acid (ACA) grown on Ag (111) by physical vapor deposition was characterized by using UHV-STM and XPS. At lower coverage, ACA molecules exist in a 2-d gas phase on the surface at room temperature. With increased coverage (>0.4 ML), ACA molecules self-organize into distinctive adlayer structures that are correlated with underlying substrate morphology. On step-free Ag (111) regions, ACA molecules form large islands in coexistence with the 2-d ACA gas. These islands are commensurate with the Ag (111) substrate, indexed as (4 0, 2 4) in matrix notation, and can exceed 100 nm in size. There are two nonequivalent ACA molecules in each unit cell. XPS core level measurements reveal a hydrogen-bonding interaction between ACA molecules, with the ring nitrogen acting as the H-bond acceptor and the carboxyl proton acting as the H-bond donor. A structural model for this phase consists of chains of ACA molecules linked by head-to-tail hydrogen bonds along the substrate [10] direction. Alternating ACA tilting angles account for the two nonequivalent ACA molecules and the observed high packing density. Completely different molecular arrangements are observed on Ag (111) surface regions roughened by a higher density of crystallographic steps (terrace widths < or = 6 nm). Pairs of ACA molecules arrange in a zigzag pattern in a (12 2, 6 5) overlayer structure with a diluted packing density. The structural model for this lower density phase consists of carboxyl-carboxyl linked ACA dimers in a flat-lying molecular orientation. PMID- 16471675 TI - Enhanced protonation of cresol red in acidic aqueous solutions caused by freezing. AB - The protonation degree of cresol red (CR) in frozen aqueous solutions at 253 or 77 K, containing various acids (HF, HCl, HNO3, H2SO4, and p-toluenesulfonic acid), sodium hydroxide, NaCl, or NH4Cl, was examined using UV/Vis absorption spectroscopy. CR, a weak organic diacid, has been selected as a model system to study the acid-base interactions at the grain boundaries of ice. The multivariate curve resolution alternating least-squares method was used to determine the number and abundances of chemical species responsible for the overlaying absorption visible spectra measured. The results showed that the extent of CR protonation, enhanced in the solid state by 2-4 orders of magnitude in contrast to the liquid solution, is principally connected to an increase in the local concentration of acids. It was found that this enhancement was not very sensitive to either the freezing rate or the type of acid used and that CR apparently established an acid-base equilibrium prior to solidification. In addition, the presence of inorganic salts, such as NaCl or NH4Cl, is reported to cause a more efficient deprotonation of CR in the former case and an enhanced protonation in the latter case, being well explained by the theory of Bronshteyn and Chernov. CR thus served as an acid-base indicator at the grain boundaries of ice samples. Structural changes in the CR molecule induced by lowering the temperature and a presence of the constraining ice environment were studied by the absorption and 1H NMR spectroscopies. Cryospheric and atmospheric implications concerning the influence of acids and bases on composition and reactivity of ice or snow contaminants were examined. PMID- 16471676 TI - Solvent effect on self-assembled structures of 3,8-bis-hexadecyloxy benzo[c]cinnoline on highly oriented pyrolytic graphite. AB - 3,8-Bis-hexadecyloxy-benzo[c]cinnoline (BBC16) self-assembled into two structures at highly oriented pyrolytic graphite (HOPG) surface: one was formed by molecules with a V-like configuration (C2v symmetry) and the other by molecules with a Z like configuration (C(s) symmetry). The self-assembled structures could be tweaked by the solvents used. In the self-assembled monolayers (SAMs) on HOPG, the BBC16 molecule adopted the V-like configuration in polar solvents and the Z like configuration in nonpolar solvents. Moreover, the solvent viscosity, solvent dissolvability of BBC16, and substrate temperature also played some roles in tuning the two-dimensional self-assembled structures. PMID- 16471677 TI - Influence of electronic properties of naphthalene compounds on contact angles. AB - Contact angles of a homologous series of naphthalene compounds on films of a fluorinated acrylate polymer (EGC-1700) deviate from an ideal pattern of contact angles. The deviations increase with the electronegativity of the constituent atoms of the liquid molecules. The results suggest that an uneven distribution of electrostatic charges over the molecules creates strong dipole moments, giving rise to fairly strong dipole-dipole and dipole-induced dipole interactions between liquid molecules and the EGC-1700 chains, which have large dipole moments. In comparison, contact angles of the same probe liquids on the films of Teflon AF 1600, which have small dipole moments, fall on a smooth curve representing the surface tension of the polymer film. PMID- 16471678 TI - Molecular chirality and charge transfer through self-assembled scaffold monolayers. AB - The effect of molecular chirality on electron transmission is explored by photoelectrochemistry. Thiol-terminated chiral scaffold molecules containing a porphyrin chromophore were self-assembled on gold surfaces to form a monolayer. Incorporation of the SAM-coated gold into an electrochemical cell and illumination with visible light generated a cathodic photocurrent. When using circularly polarized light, the photocurrent displayed an asymmetry (different magnitude of photocurrent for right versus left polarization) that changed with the molecular chirality (left- or right-handedness of the scaffold). A symmetry constraint on the electronic coupling between the porphyrin and the organic scaffold is proposed as a possible mechanism for the photocurrent asymmetry. PMID- 16471679 TI - Adsorption of bromobenzene on periodically stepped and nonstepped NiO(100). AB - Periodically stepped NiO(100) surfaces were prepared and characterized with low energy electron diffraction (LEED), Auger electron spectroscopy (AES), X-ray photoelectron spectroscopy (XPS), and temperature-programmed desorption (TPD). Two vicinal NiO(100) single-crystal samples were cut, oriented, and polished with regular, repeating monatomic steps in six-atom or seven-atom terrace widths. LEED diffraction patterns showed characteristic spot-splitting that corresponded to the appropriate terrace and step height. The nonstepped and stepped NiO(100) surfaces were exposed to bromobenzene at 130 K first to produce a molecularly adsorbed monolayer species and then, with increased exposure, a multilayer adsorbate. An additional adsorbate species, observed only on the stepped surfaces, was found to desorb at 145 K by two competing pathways. One pathway, which saturates at low coverages, leaves bromine behind on the substrate and results in dehalogenation. The other pathway yields molecular desorption at 145 K, but is only observed in detectable amounts after the dehalogenation pathway is saturated. On both stepped and nonstepped NiO(100) substrates, adsorbed bromine resulting from dehalogenation processes appears as nickel bromide, determined by the Br 3p XPS data. PMID- 16471680 TI - Energy transfer at a gas-liquid interface: kinematics in a prototypical system. AB - A detailed characterization of collisional energy transfer at a liquid surface not only provides a framework for the interpretation of experimental studies but also affords insight into energy feedback mechanisms that may be important in multiphase combustion processes. We address this problem by performing simulations of a prototypical Lennard-Jones system, investigating the dependence of the energy transfer and incident-atom trapping probability on the liquid temperature, on the mass and angle of incidence of the impinging atom, and on the strength of the gas-liquid interaction. In general, in agreement with the results of experiments, these calculations point to the dominance of kinematic effects in determining the gross energy transfer, but they also attest to the important role played by surface roughening in the enhancement of energy transfer that accompanies an increase in the liquid temperature. PMID- 16471681 TI - Electric double layers with electrolyte mixtures: integral equations theories and simulations. AB - A study of a planar electric double layer (EDL) in the presence of mixtures of electrolyte is presented. In particular, results from the Hyper-Netted-Chain/Mean Spherical-Approximation (HNC/MSA) theory are compared with Monte Carlo (MC) simulations. In this way, the charge inversion induced by mixtures of multivalent and monovalent counterions is probed. Since overcharging phenomena in nature emerge under such conditions, the role of ion-ion correlations in the EDL appears as a crucial point in this kind of study. Unlike previous related works, a realistic hydrated ion size is used in the HNC/MSA calculations and simulations. In this way, a qualitative agreement between the results obtained from the theory and MC simulations is found. However, some discrepancies arise when the charge inversion is expected to be more noticeable, namely at high surface charges and/or elevated concentrations of multivalent electrolytes. Such differences are explained in terms of an overestimation of the charge inversion by the integral equation (IE) formalism. PMID- 16471682 TI - Electronic properties of Si surfaces and side reactions during electrochemical grafting of phenyl layers. AB - The electrochemical grafting process of 4-nitrobenzene and 4-methoxybenzene (anisole) from diazonium salt solutions has been investigated in situ by monitoring the current density, the band bending, and the nonradiative surface recombination during grafting at different potentials and different concentrations of the diazonium salt in the solution. Ex situ infrared spectroscopic ellipsometry has been used to inspect the Si surface species before and after the grafting process. The band bending decreases with either increasing concentration of diazonium salt or when the redox potential of the diazonium compound (anisole) is nearer to the competing H+/H2 couple. The surface recombination increases at more cathodic potentials if an electron donor group is present at the phenyl ring (nitrobenzene) and vice versa for the electron acceptor group (anisole). The influence of side reactions can be reduced by use of moderate concentration and moderate or strong cathodic potential, depending on the redox potential of the diazonium compound. PMID- 16471683 TI - Structural model of silica nanowire assembled from a highly stable (SiO2)8 unit. AB - The ground-state structures of silica clusters (SiO2)n for n = 1-8 were studied by performing calculations at the B3LYP/6-311+G(d) level of density functional theory. The results indicate that the growth mode of a silica nanowire based on small silica clusters may change at different wire lengths. A linear chain might be assembled from the smallest clusters of rhombic two-membered ring (2MR) with n < or = 5, while the growth motif changes at n = 6 into a more compact form composed of three-membered-rings (3MRs). The 3MR-containing structures become energetically favorable configurations for even longer silica clusters. In particular, the closed molecular ring consisting of 3MRs at n = 8 (i.e., (SiO2)8) with a high symmetry shows extreme energetic stability and relatively high chemical reactivity and thus is considered to be an important building block to assemble into silica nanowires. The relative stability of so-assembled silica nanowires were evaluated and compared with the models of silica nanowires in the literature. PMID- 16471684 TI - First principles study of adsorption and dissociation of CO on W(111). AB - The adsorption and dissociation of carbon monoxide on the W(111) surface is studied with density functional theory. The CO molecule is found to adsorb in end on configurations (alpha states) and inclined configurations (beta states). The dissociation of the most strongly bound beta state CO is found to have an activation energy of about 0.8 eV, which is lower than the energy required to desorb CO molecularly from the surface. The diffusion of CO and O on W(111) is predicted to be facile at room temperature, whereas C atoms are virtually immobile up to approximately 600 K, according to our calculations. Preadsorbed carbon atoms are shown to prevent the dissociation of CO by blocking the most strongly bound beta state adsorption site and by blocking the dissociation pathway. We predict that dissociation of CO on W(111) is a self-poisoning process. PMID- 16471685 TI - Global fluid phase equilibria and critical phenomena of selected mixtures using the crossover soft-SAFT equation. AB - We present here the extension of the crossover soft-statistical associating fluid theory (soft-SAFT) equation of state to mixtures, as well as some illustrative applications of the methodology to mixtures of particular scientific and technological interest. The procedure is based on White's work (White, J. A. Fluid Phase Equilib. 1992, 75, 53) from the renormalization group theory, as for the pure fluids, with the isomorphism assumption applied to the mixtures. The equation is applied to three groups of mixtures: selected mixtures of n-alkanes, the CO2/n-alkane homologous series, and the CO2/1-alkanol homologous series. The crossover equation is first applied to the pure components of the mixtures, CO2 and the 1-alkanol family, while an available correlation is used for the molecular parameters of the n-alkane series (Llovell et al. J. Chem. Phys 2004, 121, 10715). A set of transferable molecular parameters is provided for the 1 alkanols series; these are accurate for the whole range of thermodynamic conditions. The crossover soft-SAFT equation is able to accurately describe these compounds near to and far from the critical point. The theory is then used to represent the phase behavior and the critical phenomena of the selected mixtures. We use binary interaction parameters xi and eta for dissimilar mixtures. These parameters are fitted at some particular conditions (one subcritical temperature or binary critical data) and used to predict the behavior of the mixture at different conditions (other subcritical conditions and/or critical conditions). The equation is able to capture the continuous change in the critical behavior of the CO2/n-alkane and the CO2/1-alkanol homologous series as the chain length of the second compound increases. Excellent agreement with experimental data is obtained, even in the most nonideal cases. The new equation is proved to be a powerful tool to study the global phase behavior of complex systems, as well as other thermodynamic properties of very challenging mixtures. PMID- 16471686 TI - Testing assumptions about solute concentration dependence in liquid crystal NMR. AB - The NMR spectra of four solutes, used as probes of liquid crystal orientational order, were analyzed. For each solute, samples were prepared at different solute concentrations, and the concentration dependence was used to extrapolate zero concentration properties. The mean-field (Maier-Saupe) model when applied to solutes neglects solute-solute interactions and assumes all solutes in a mixed solute sample see the same average environment. The first assumption is only valid as one approaches zero concentration, while experiments are typically carried out at concentrations between 0 and 10 mol %. The solute concentration dependence has in the past been "scaled out" using an internal solute reference as an orientational standard. We measured the concentration dependence of the orientational order parameter and calculate the corresponding interaction energies based on a mean-field interaction potential for a solute. We find agreement at the 3% level between experiments for different solutes while using (i) the zero-concentration values as solute-dependent orientational references and (ii) scaling to either order parameters or interaction energies; these two scalings gave equivalent but not identical results. We find, too, that errors inherent in the experiment and the calculations will limit attempts to refine the theory to push the comparisons beyond the 2% level. PMID- 16471687 TI - Calculation of phase coexistence properties and surface tensions of n-alkanes with grand-canonical transition-matrix monte carlo simulation and finite-size scaling. AB - Grand-canonical transition-matrix Monte Carlo is combined with configurational bias and expanded ensemble Monte Carlo techniques to obtain saturated densities and vapor pressures of select n-alkanes. Surface tension values for butane, hexane, and octane are also computed via the finite-size scaling method of Binder. The exponential-6 model of Errington and Panagiotopoulos is used to describe the molecular interactions. The effect of the number of configurational bias trial conformations on the efficiency of phase equilibra calculations is studied. We find that a broad range of trial conformation numbers give reasonable performance, with the optimal value increasing with decreasing temperature for a fixed chain length. Phase coexistence properties are in good agreement with literature values and are obtained with very reasonable computing resources. Similar to other recently developed n-alkane force fields, the exponential-6 model overestimates the surface tension relative to experimental values. Statistical uncertainties for coexistence properties obtained with the current approach are relatively small compared to existing methods. PMID- 16471688 TI - A simple quantum statistical thermodynamics interpretation of an impressive phase diagram pressure shift upon (H/D) isotopic substitution in water + 3 methylpyridine. AB - In a previous work (J. Phys. Chem. B 2003, 107, 9837), we reported liquid-liquid phase splitting at negative pressures in mixtures of H2O + D2O + 3-methylpyridine (3-MP) at the limit of pure H2O as the solvent, thus extending for the first time the L-L phase diagrams to this metastable region. We showed that there is an intimate relation between pressure and solvent deuterium content. Isotopic substitution (H/D) in water provokes subtle entropic effects that, in turn, trigger a significant pressure shift, opening a pressure-wide miscibility window of as much as 1600 bar. Isotope effects are quantum in origin. Therefore, a model that is both pressure-dependent and considers quantization constitutes a necessary tool if one wishes to fully describe the p, T, x critical demixing in these systems. In the current work, the statistical-mechanical theory of isotope effects is combined with a compressible pressure-dependent model. This combination enabled us to predict successfully the overall L-L phase diagram via differences in the vibrational mode frequencies of water on its transfer from the pure state to that of dilution in 3-MP: each of the three librational modes undergo a calculated red-shift of -(250 +/- 30) cm(-1), while the overall internal frequencies contribution is estimated as a total +(400 +/- 25) cm(-1) blue-shift. PMID- 16471689 TI - Comparative study of the energy transfer kinetics in artificial BChl e aggregates containing a BChl a acceptor and BChl e-containing chlorosomes of Chlorobium phaeobacteroides. AB - Chlorosomes are the light-harvesting organelles of green bacteria, containing mainly special bacteriochlorophylls (BChls) carrying a 3(1)-hydroxy side chain. Artificial aggregates of BChl c, d, and e have been shown to resemble the native chlorosomes in many respects. They are therefore seen as good model systems for understanding the spectroscopic properties of these antenna systems. We have investigated the excitation energy transfer in artificial aggregates of BChl e, containing small amounts of BChl a as an energy acceptor, using steady-state and time-resolved fluorescence. Global analysis of the kinetic data yields two lifetimes attributable to energy transfer: a fast one of 12-20 ps and a slower one of approximately 50 ps. For comparison, BChl e-containing native chlorosomes of Chlorobium phaeobacteroides and chlorosomes in which the energy acceptor had been degraded by alkaline treatment were also studied. A similar behavior is seen in both the artificial and the natural systems. The results suggest that the artificial aggregates of BChls have a potential as antenna systems in future artificial photonic devices. PMID- 16471690 TI - Interpreting the catalytic voltammetry of an adsorbed enzyme by considering substrate mass transfer, enzyme turnover, and interfacial electron transport. AB - Redox active enzymes can be adsorbed onto electrode surfaces to catalyze the interconversion of oxidized and reduced substrates in solution, driven by the supply or removal of electrons by the electrode. The catalytic current is directly proportional to the rate of enzyme turnover, and its dependence on the electrode potential can be exploited to define both the kinetics and thermodynamics of the enzyme's catalytic cycle. However, observed electrocatalytic voltammograms are often complex because the identity of the rate limiting step changes with the electrode potential and under different experimental conditions. Consequently, extracting mechanistic information requires that accurate models be constructed to deconvolute and analyze the observed behavior. Here, a basic model for catalysis by an adsorbed enzyme is described. It incorporates substrate mass transport, enzyme kinetics, and interfacial electron transport, and it accurately reproduces experimentally recorded voltammograms from the oxidation of NADH by subcomplex Ilambda (the hydrophilic subcomplex of NADH:ubiquinone oxidoreductase), under a range of conditions. Mass transport is imposed by a rotating disk electrode and described by the Levich equation. Interfacial electron transport is controlled by the electrode potential and characterized by a dispersion of rate constants, according to the model of Leger and co-workers. Here, the Michaelis-Menten equation is used for the enzyme kinetics, but our methodology can also be readily applied to derive and apply analogous equations relating to alternative enzyme mechanisms. Therefore, our results are highly relevant to the interpretation of electrocatalytic voltammograms for adsorbed enzymes in general. PMID- 16471691 TI - The effect of salt on protein chemical potential determined by ternary diffusion in aqueous solutions. AB - We use accurate thermodynamic derivatives extracted from high-precision measurements of the four volume-fixed diffusion coefficients in ternary solutions of lysozyme chloride in aqueous NaCl, NH4Cl, and KCl at pH 4.5 and 25 degrees C to (a) assess the relative contributions of the common-ion and nonideality effects to the protein chemical potential as a function of salt concentration, (b) compare the behavior of the protein chemical potential for the three salts, which we found to be consistent with the Hofmeister series, and (c) discuss our thermodynamic data in relation to the dependence of the protein solubility on salt concentration. The four diffusion coefficients are reported at 0.6 mM lysozyme chloride and 0.25, 0.5, 0.9, 1.2, and 1.5 M KCl and extend into the protein-supersaturated region. The chemical potential cross-derivatives are extracted from diffusion data using the Onsager reciprocal relation and the equality of molal cross-derivatives of solute chemical potentials. They are compared to those calculated previously from diffusion data for lysozyme in aqueous NaCl and NH4Cl. We estimate the effective charge on the diffusing lysozyme cation at the experimental concentrations. Our diffusion measurements on the three salts allowed us to analyze and interpret the four diffusion coefficients for charged proteins in the presence of 1:1 electrolytes. Our results may provide guidance to the understanding of protein crystallization. PMID- 16471692 TI - Modeling of hydrogen bonds in monohydrated 2,4-dithiothymine: an ab initio and AIM study. AB - Twelve tautomers of 2,4-dithiothymine are calculated at the MP2/6-31+G(d) level, and the most stable one is referred to the di-keto form (P12). Then four H-bonded complexes between P12 and water are optimized at the MP2/6-31+G(d) level of theory. The calculation of vibrational frequencies and natural bond orbital analysis are also carried out at the same level to investigate the hydrogen bonds involved in all the systems. Within all the four complexes, three types of hydrogen bonds are formed, in which the O-H...S and N-H...O bonds are the normal bonds with the X-H bond elongation and red shift of the corresponding stretch frequencies, while the C-H...O interaction is an improper, blue-shifting hydrogen bond accompanied with the contraction of the C-H bond and a blue shift of the C-H stretch frequency. The topological properties are investigated with the atoms-in molecules (AIM) theory. The NMR chemical shielding for the isolated and the four monohydrated 2,4-dithiothymine are calculated using the "gauge-including atomic orbital" (GIAO) method. The 1H chemical shifts are influenced by the formation of hydrogen bonds. PMID- 16471693 TI - Conformational polymorphism of the PrP106-126 peptide in different environments: a molecular dynamics study. AB - Extensive molecular dynamic simulations (approximately 240 ns) have been used to investigate the conformational behavior of PrP106-126 prion peptide in four different environments (water, dimethyl sulfoxide, hexane, and trifluoroethanol) and under both neutral and acidic conditions. The conformational polymorphism of PrP106-126 in solution observed in the simulations supports the role of this fragment in the structural transition of the native to the abnormal form of prion protein in response to changes in the local environmental conditions. The peptide in solution is primarily unstructured. The simulations show an increased presence of helical structure in an apolar solvent, in agreement with the results from circular dichroism spectroscopy. In water solution, beta-sheet elements were observed between residues 108-112 and either residues 115-121 or 121-126. An alpha-beta transition was observed under neutral conditions. In DMSO, the peptide adopted an extended conformation, in agreement with nuclear magnetic resonance experiments. PMID- 16471694 TI - Direct quantification of intraparticle protein diffusion in chromatographic media. AB - Diffusion coefficients of proteins in chromatographic media are important parameters for the rational design of stationary phases and purification schemes. In contrast to free diffusion, intraparticle diffusion is hindered by the porous structure of the media. Direct intraparticle diffusion analysis (IDA) is a novel approach for the determination of intraparticle protein diffusion coefficients. IDA is based on the evaluation of spatially and temporally resolved intraparticle concentration profiles. To prevent adsorption and to study diffusion only, the chromatographic media are investigated in underivatized form. With IDA, intraparticle concentration profiles are measured in a microcolumn by confocal laser scanning microscopy (CLSM). From this dynamic data, the diffusion coefficients are determined by parameter estimation, using a spheric diffusion model. The boundary condition is given by the measured protein concentration in the bulk phase. IDA is applied to determine intraparticle diffusion coefficients of seven different proteins in Sepharose 6 FF. The results show excellent congruence of experimental data and simulation results. Moreover, the determined diffusion coefficients lie well within the range of data published in the literature. Given that the material in question allows optical analysis, IDA is a general approach for studying protein diffusion in porous particles and is easily adapted to different proteins, solution conditions and stationary phases. PMID- 16471695 TI - Binding of phosphinate and phosphonate inhibitors to aspartic proteases: a first principles study. AB - Phosphinate and phosphonate derivatives are potent inhibitors of aspartic proteases (APs). The affinity for the enzyme might be caused by the presence of low barrier hydrogen bonds between the ligand and the catalytic Asp dyad in the cleavage site. We have used density functional theory calculations along with hybrid quantum mechanics/molecular mechanics Car-Parrinello molecular dynamics simulations to investigate the hydrogen-bonding pattern at the binding site of the complexes of human immunodeficiency virus type-1 AP and the eukaryotic endothiapepsin and penicillopepsin. Our calculations are in fair agreement with the NMR data available for endothiapepsin (Coates et al. J. Mol. Biol. 2002, 318, 1405-1415) and show that the most stable active site configuration is the diprotonated, negatively charged form. In the viral complex both protons are located at the catalytic Asp dyad, while in the eukaryotic complexes the proton shared by the closest oxygen atoms is located at the phosphinic/phosphonic group. PMID- 16471697 TI - All electron quantum chemical calculation of the entire enzyme system confirms a collective catalytic device in the chorismate mutase reaction. AB - To elucidate the catalytic power of enzymes, we analyzed the reaction profile of Claisen rearrangement of Bacillus subtilis chorismate mutase (BsCM) by all electron quantum chemical calculations using the fragment molecular orbital (FMO) method. To the best of our knowledge, this is the first report of ab initio-based quantum chemical calculations of the entire enzyme system, where we provide a detailed analysis of the catalytic factors that accomplish transition-state stabilization (TSS). FMO calculations deliver an ab initio-level estimate of the intermolecular interaction between the substrate and the amino acid residues of the enzyme. To clarify the catalytic role of Arg90, we calculated the reaction profile of the wild-type BsCM as well as Lys90 and Cit90 mutant BsCMs. Structural refinement and the reaction path determination were performed at the ab initio QM/MM level, and FMO calculations were applied to the QM/MM refined structures. Comparison between three types of reactions established two collective catalytic factors in the BsCM reaction: (1) the hydrogen bonds connecting the Glu78-Arg90 substrate cooperatively control the stability of TS relative to the ES complex and (2) the positive charge on Arg90 polarizes the substrate in the TS region to gain more electrostatic stabilization. PMID- 16471696 TI - Mechanism of dihydroneopterin aldolase: a molecular dynamics study of the apo enzyme and its product complex. AB - Dihydroneopterin aldolase (DHNA), an enzyme in the pathway that generates folic acid in bacteria, is investigated by a series of molecular dynamics simulations in its free form and complexed with its product, 6-hydroxymethyl-7,8 dihydropterin (HP). The active sites in DHNA are formed at the interface between pairs of protomers in this octameric protein. On the basis of root-mean-square deviation and root-mean-square fluctuation analyses of the trajectories, which take advantage of the presence of eight active sites, flexible regions of the apo protein surrounding the active site are identified and, upon binding HP, show that the active site is rigidified. Specific residues, associated with binding and the catalytic mechanism of DHNA, are associated with these flexible regions, and their interactions with HP account for most of the binding energy. A Principal Component Analysis shows rigidification of DHNA upon HP binding and that only a few modes of motion capture most of the atomic fluctuations in both apo and HP-bound forms. HP is pushed out of the active site in a series of simulations with different restrained positions between HP and DHNA to obtain a view of the exit pathway and energetic barrier to product release. The chosen pathway leads to a minimal disturbance of the system and provides a barrier consistent with the experimentally determined rate of product release. An analysis of the various components that contribute to the exit path energy and entropy provides insight into the energy-entropy compensation for product release. PMID- 16471698 TI - Thermodynamics of buried water clusters at a protein-ligand binding interface. AB - The structure of the complex of cyclophilin A (CypA) with cyclosporin A (CsA, 1) shows a cluster of four water molecules buried at the binding interface, which is rearranged when CsA is replaced by (5-hydroxynorvaline)-2-cyclosporin (2). The thermodynamic contributions of each bound water molecule in the two complexes are explored with the inhomogeneous fluid solvation theory and molecular dynamics simulations. Water (WTR) 133 in complex 1 contributes little to the binding affinity, while WTR6 and 7 in complex 2 play an essential role in mediating protein-ligand binding with a hydrogen bond network. The calculations reveal that the rearrangement of the water molecules contributes favorably to the binding affinity, even though one of them is displaced going from ligand 1 to 2. Another favorable contribution comes from the larger protein-ligand interactions of ligand 2. However, these favorable contributions are not sufficient to overcome the unfavorable desolvation free energy change and the conformational entropy of the hydroxylpropyl group of ligand 2 in the complex, leading to a lower binding affinity of ligand 2. These physical insights may be useful in the development of improved scoring functions for binding affinity prediction. PMID- 16471700 TI - Investigations on the kinetics of electron transfer reactions in magnetic fields. AB - There is a controversial debate if a magnetic field can influence the rate of electron transfer (ET) reactions. In this paper, we report kinetic measurements of the ET rate constants for the redox couples [IrCl6]2-/[IrCl6]3-, [Fe(CN)6]3 /[Fe(CN)6]4-, and [Fe(H2O)6]3+/[Fe(H2O)6]2+ in magnetic fields up to 1 T. To reduce effects arising from magnetically induced mass transport (magnetohydrodynamic effect), disk microelectrodes with a diameter of 50 microm were used in potentiodynamic (cyclic and linear sweep voltammetry) and in electrochemical impedance spectroscopy experiments. None of the investigated redox couples showed a magnetic field effect on the ET rate constant. PMID- 16471699 TI - Solvation model based on order parameters and a fast sampling method for the calculation of the solvation free energies of peptides. AB - An analytical solvation model is proposed as a function of an order parameter, which represents the local arrangement of water molecules in the first solvation shell of peptide atoms. The model is combined with a fast sampling method, rotational isomeric state Monte Carlo, to sample efficiently the torsional degrees of freedom on a peptide backbone. This order parameter solvation model is shown to reproduce without ad hoc fitting parameters the solvation free energies of single amino acids and tripeptides with slightly better accuracy than the generalized Born model but with several orders of magnitude improvement in efficiency. This method is a potential candidate for efficiently and accurately tackling some important issues in biophysical chemistry that are related to solvation, for example, protein folding, ligand binding, etc. Our results also present fundamental new insights into solvation. Specifically, the local water geometry, represented in this work by a properly defined order parameter, carries the majority, if not all, of the energetic information of solvation, including solute-solvent interactions and solvent reorganization in the presence of the solute. PMID- 16471701 TI - Electrophoresis of a spherical dispersion of polyelectrolytes in a salt-free solution. AB - The electrophoretic behavior of a spherical dispersion of polyelectrolytes of arbitrary concentration is analyzed theoretically under a salt-free condition, that is, the liquid phase contains only counterions which come from the dissociation of the functional groups of polyelectrolytes. We show that, in general, the surface potential of a polyelectrolyte increases nonlinearly with its surface charge. A linear relation exists between them, however, when the latter is sufficiently small; and the more dilute the concentration of polyelectrolytes, the broader the range in which they are linearly correlated. If the amount of surface charge is sufficiently large, counterion condensation occurs, and the rate of increase of surface potential as the amount of surface charge increases declined. Also, it leads to an inverse in the perturbed potential near the surface of a polyelectrolyte, and its mobility decreases accordingly. For a fixed amount of surface charge, the lower the concentration of polyelectrolytes and/or the lower the valence of counterions, the higher the surface potential. The qualitative behavior of the mobility of a polyelectrolyte as the amount of its surface charge varies is similar to that of its surface charge. PMID- 16471702 TI - Domain formation in lipid bilayers probed by two-dimensional infrared spectroscopy. AB - Two-dimensional infrared (2D-IR) spectroscopy has been used to probe structure and dynamics in binary sphingomyelin/phospholipid liposomes. The liposomes consist of 1-palmitoyl-2-linoleyl phosphatidylcholine (PLPC) and sphingomyelin (SPM) in the ratio 1:1. The diagonal part of the 2D-IR spectra shows two bands which are due to amide I of SPM and to the carbonyl moieties of PLPC. The diagonal components of the 2D-IR spectra reveal a difference in the molecular dynamics. The presence of off-diagonal cross-peaks indicates the occurrence of intermolecular structural correlation. The intensity of the cross-peaks is consistent with segregation of two lipid components into PLPC and SPM molecular domains. PMID- 16471703 TI - Thermally and molecularly stimulated relaxation of hot phonons in suspended carbon nanotubes. AB - The high-bias electrical transport properties of suspended metallic single-walled carbon nanotubes (SWNTs) are investigated at various temperatures in vacuum, in various gases, and when coated with molecular solids. It is revealed that nonequilibrium optical phonon effects in suspended nanotubes decrease as the ambient temperature increases. Gas molecules surrounding suspended SWNTs assist the relaxation of hot phonons and afford enhanced current flow along nanotubes. Molecular solids of carbon dioxide frozen onto suspended SWNTs quench the nonequilibrium phonon effect. The discovery of strong environmental effects on high current transport in nanotubes is important to high performance nanoelectronics applications of 1D nanowires in general. PMID- 16471704 TI - Site affinity effects upon charge injection into siloxane-based monolayers. AB - Submolecular electrical information is successfully derived by applying element specific, chemically resolved electrical measurements to a covalently bound stilbazole-based monolayer on a silicon substrate. Pronounced affinity effects are found in the response of adjacent atomic sites to external charge injection, accompanied by intramolecular polarization variations. These noncontact electrical read-out capabilities may provide a first entry toward the realization of organic devices based on submolecular electrical units. PMID- 16471705 TI - Does magnetic treatment of water change its properties? AB - Some properties and functions of water treated under magnetic field were examined. No change in properties of pure water distilled from ultrapure water in vacuum was observed by magnetic treatment. However, when the same magnetic treatment was carried out after the distilled water was exposed to O2, water properties such as vibration modes and electrolytic potential were changed. The degree of magnetic treatment effect on water was quantitatively evaluated by contact angle. PMID- 16471706 TI - Morphology of poly(ethylene oxide) dissolved in a room temperature ionic liquid: a small angle neutron scattering study. AB - Solutions of deuterated poly(ethylene oxide) (d-PEO) in 1-butyl-3-methyl imidazolium tetrafluoroborate ([bmim][BF4]), a prototype room-temperature ionic liquid (RTIL), have been studied at room temperature over a range of polymer concentrations, using small angle neutron scattering (SANS), characterizing the conformation of PEO dissolved in RTILs. [bmim][BF4] behaves as a good solvent for d-PEO, which organizes in this solvent in non entangled random coils. These findings will help in optimizing the designing of microemulsions in these potentially environmentally friendly solvents. PMID- 16471707 TI - Fabrication and photoluminescent properties of heteroepitaxial ZnO/Zn0.8Mg0.2O coaxial nanorod heterostructures. AB - ZnO/Zn0.8Mg0.2O coaxial nanorod heterostructures were prepared by employing catalyst-free metal-organic vapor-phase epitaxy, and their structural and photoluminescent (PL) properties were investigated using transmission electron microscopy (TEM) and temperature-dependent PL spectroscopy. TEM images show that ZnO/Zn0.8Mg0.2O layers were epitaxially grown on the entire surfaces of the ZnO nanorods and the ZnO nanorod diameters as a core material were as small as 9 +/- 2 nm. A dominant PL peak was observed at 3.316 eV, from room-temperature PL spectra of ZnO/Zn0.8Mg0.2O coaxial nanorod heterostructures with ZnO core diameters of 9 nm, indicating a PL blue shift of 30 meV, which resulted from a quantum confinement effect along the radial direction in ZnO nanorods. Furthermore, temperature-dependent PL properties of the coaxial nanorod heterostructures were investigated, showing much higher PL intensity for the coaxial nanorod heterostructures than that of bare ZnO nanorods at room temperature. The origin of the enhanced PL intensity and reduced thermal quenching for the coaxial nanorod heterostructures is also discussed. PMID- 16471709 TI - Purification of boron nitride nanotubes through polymer wrapping. AB - An effective method was proposed to remove obstinate boron nitride phase impurities in boron nitride nanotubes (BNNTs). The method is based on strong interactions between BNNTs and a conjugated polymer wrapping them and significant weight and size difference between BNNTs and impurities. The as-grown samples and purified samples were compared through detailed characterization, using scanning electron microscopy, transmission electron microscopy, and Raman and Fourier transformed infrared spectroscopy. The results reveal that impurities are effectively removed and resultant BNNTs possess perfect crystallization. PMID- 16471708 TI - Ultrafast laser studies of the photothermal properties of gold nanocages. AB - Au nanocages were synthesized via a galvanic replacement reaction. The extinction peak of these hollow structured particles is shifted into the near-IR compared with the Ag nanocube templates. Energy transfer from the Au nanocages into the surrounding environment (water) as well as the coherently excited vibrational modes of the nanocages were studied by femtosecond pump-probe spectroscopy. The time scale for energy relaxation was found to increase with the size of the particles, with the relaxation time being independent of the laser intensity. The time scales for relaxation are comparable to those for solid spherical gold particles and are consistent with energy relaxation being controlled by heat dissipation in the solvent. The period of the coherently excited vibrational mode is proportional to the dimensions of the nanocages. Intensity-dependent measurements show that in solution the nanocages maintain their integrity up to lattice temperatures of 1100 +/- 100 K. PMID- 16471710 TI - Changes in electrochemical and optical properties of oligoalkylthiophene film induced by bipolaron formation. AB - An unusual electrochemical response is found for oligoalkylthiophene (13T) films in acetonitrile. A pristine 13T film exhibits a stable redox wave in cyclic voltammogram when the anodic potential limit is below 0.6 V, whereas the redox peaks shift clearly to the negative side by ca. 0.1 V once the 13T film experiences a potential beyond 0.7 V. After this electrochemical stimulus is imposed on a pristine 13T film, the absorption and emission spectra change their features. These findings, along with the measurements with varying thicknesses of 13T films, suggest that a pristine 13T film is a mixture of two phases to be oxidized at separate potentials and that one phase can be switched to the other by the potential stimulation. A plausible model proposed for explaining these observations assumes that a key parameter characteristic of the two phases is a conformation of 13T molecules in the solid state and a change in conformation, leading to the shift of the redox potentials and the change of optical properties, is induced by electrochemical generation of bipolarons on the half units of the oligomers in the pristine 13T film. PMID- 16471711 TI - Intramolecular electron transfer in cofacially pi-stacked fluorenes: evidence of tunneling. AB - The one-electron reduction of neutral pi-stacked di- and trifluorenes (F-2 and F 3) in HMPA, where ion association is absent, results in the formation of anion radicals in which the odd electron resides predominantly on just one of the external fluorene moieties, as established by EPR spectroscopy. However, in the case of tetrafluorene, introduction of a single electron leads to a kinetically controlled anion radical F-4(int)*- in which the odd electron undergoes rapid exchange between two central fluorene rings, where the anionic charge is partially shielded from solvation due to the presence of external fluorene rings. On a time scale of minutes, anion radical F-4(int)*- converts to a thermodynamically stabilized anion radical F-4(ext)*-, with the electron exhibiting coupling from the protons on an external fluorene moiety. The charge and spin residing on an external moiety allow efficient solvation of the anionic charge. A similar fast exchange of a single electron (probably with the involvement of quantum mechanical tunneling) among three and four internal fluorene moieties is initially observed via EPR spectroscopy in the penta- and hexafluorene derivatives, F-5 and F-6, respectively. PMID- 16471712 TI - Dielectrophoresis of surface conductance modulated single-walled carbon nanotubes using catanionic surfactants. AB - Dielectrophoresis has received considerable attention for separating nanotubes according to electronic types. Here we examine the effects of surface conductivity of semiconducting single walled carbon nanotubes (SWNT), induced by ionic surfactants, on the sign of dielectrophoretic force. The crossover frequency of semiconducting SWNT increases rapidly as the conductivity ratio between the particle and medium increases, leading to an incomplete separation of ionic surfactant suspended SWNT at an electric field frequency of 10 MHz. To reduce the conductivity ratio, the surface charge of SWNT is neutralized by an equimolar mixture of anionic surfactant sodium dodecyl sulfate (SDS) and cationic surfactant cetyltrimethylammonium bromide (CTAB), resulting in negative dielectrophoresis of semiconducting species at 10 MHz. A comparative Raman spectroscopy study shows a nearly complete separation of metallic SWNT. PMID- 16471713 TI - Ultrasonically induced Au nanoprisms and their size manipulation based on aging. AB - A simple sonochemical approach was developed for the synthesis of single-crystal Au nanoprisms with triangular or hexagonal shape, 30-40 nm planar dimension, and 6-10 nm thickness in an ethylene glycol solution. It has been shown that ethylene glycol, the surfactant poly(vinylpyrrolidone), and ultrasonic irradiation play important roles in the formation of Au nanoprisms. The subsequent aging induces the growth of Au nanoprisms, which is attributed to the existence of residual reduced Au degrees atoms and Au clusters in the solution. Further growth of Au nanoprisms could be realized by adding a given amount of HAuCl4 to the irradiated and aged Au nanodisk colloidal solution, which provides a simple and an effective method to synthesize Au nanoprisms with different sizes. Au nanoprisms with planar sizes from 70 to 400 nm have been successfully synthesized based on this strategy. PMID- 16471714 TI - Magnetic iron oxide nanoparticles for biorecognition: evaluation of surface coverage and activity. AB - Modifying the surfaces of magnetic nanoparticles (MNPs) by the covalent attachment of biomolecules will enable their implementation as contrast agents for magnetic resonance imaging or as media for magnetically assisted bioseparations. In this paper we report both the surface coverage and the activity of IgG antibodies on MNPs. The antibodies were immobilized on gamma Fe2O3 nanoparticles by conventional methods using aminopropyltriethoxy silane and subsequent activation by glutaraldehyde. Novel fluorescence methods were used to provide a quantitative evaluation of this well-known approach. Our results show that surface coverage can be stoichiometrically adjusted with saturated surface coverage occurring at approximately 36% of the theoretical limit. The saturated surface coverage corresponds to 34 antibody molecules bound to an average-sized MNP (32 nm diameter). We also show that the immobilized antibodies retain approximately 50% of their binding capacity at surface-saturated levels. PMID- 16471715 TI - Highly active TiO2N photocatalysts prepared by treating TiO2 precursors in NH3/ethanol fluid under supercritical conditions. AB - N-doped TiO2 photocatalysts were prepared by pretreating the TiO2 precursor in NH3/ethanol fluid under supercritical conditions, denoted as TiO2N(SC). In contrast to the TiO2N(DC), obtained via direct calcination in which the N dopants were mainly present in the form of surface adsorbed NH3 molecules, most N dopants in the TiO2N(SC) were present in O-Ti-N and N-Ti-N nitrides, as confirmed by either the X-ray photoelectron spectroscopy (XPS) and or the Fourier transform infrared (FTIR) spectra. During liquid-phase oxidative degradation of phenol under irradiation with UV light characteristic of 365 nm, the TiO2N(SC) exhibited much higher activity than either the TiO2N(DC) or the TiO2(SC), i.e., the undoped TiO2 obtained under SCs. According to various characterizations including X-ray diffraction, transmission electron microscopy, FTIR, Brunauer-Emmett-Teller, XPS, and UV-vis diffuse reflectance spectra, the higher activity of the TiO2N(SC) could be attributed to its higher surface area, larger pore volume, well crystallized anatase, and stronger absorbance of light with longer wavelength. Meanwhile, the OH species resulted from the nitridation of TiO2 could supply more HO* radicals, which were considered as powerful oxidants during phenol degradation. Furthermore, the electron-deficient nitrogen atoms in O-Ti-N nitrides could also account for the higher activity since it could inhibit the recombination between the photoinduced electrons and holes by capturing the photoinduced electrons. The activity of the TiO2N(SC) first increased and then decreased with the increase of the N-content. The TiO2N(SC)-1 with N/Ti molar ratio of 1.73% exhibited maximum activity, which was even much higher than P-25. PMID- 16471716 TI - Preparation and third-order optical nonlinearity of self-assembled chitosan/CdSe ZnS core-shell quantum dots multilayer films. AB - The self-assembled chitosan CdSe quantum dots (QDs) and chitosan CdSe-ZnS core shell QDs films have been prepared by using layer-by-layer electrostatic technique. The well-ordered nanostructure and the layer-by-layer deposition of the QDs are revealed by AFM and exciton absorption spectra, respectively. The optical nonlinearity of the composite films were studied by using Z-scan technique with femtosecond pulses at the wavelength of 790 nm, the value of third order susceptibility of core-shell QDs are measured to be about 1.1 x 10(-8) esu, which is about 200% larger than that of CdSe QDs of 5.3 x 10(-9) esu. This has potential applications in all-optical switches in optical information processing. PMID- 16471717 TI - Visible-light-induced water-splitting in channels of carbon nanotubes. AB - The visible-light-induced split of water confined in channels of single-walled carbon nanotubes (SWNTs) was experimentally studied. Arc-discharging synthesized SWNTs were used to adsorb water vapor and then were irradiated in a vacuum by using light from a camera flash. It was found that a great amount of hydrogen rich gases could be repeatedly produced under several rapid flashes of light, occasionally accompanying evident charge emission phenomena. A quantitative method was developed to estimate the relative amount of gas components on the basis of the data acquired with an ion gauge and a quadrupole mass spectrometer. The results indicated that hydrogen occupied about 80 mol % of the photogenerated gases, with other components such as carbon oxides, helium, methane and trace of ethane, and the total gas yield in one flash (0.1-0.2 J/cm2, 8 ms) reached 400 900 ppm of the mass of the SWNTs. Such a yield could be repeatedly obtained in serial flashings until the adsorbed water was depleted, and then, by sufficiently adsorbing water vapor again, the same phenomena could be reproduced. PMID- 16471718 TI - Phase equilibria and transition mechanisms in high-pressure AgCl by ab initio methods. AB - The theoretical study of pressure-driven phase transformations by means of ab initio quantum mechanical methods, in the frame of the extended Landau approach, is considered. A specific application to AgCl is presented: the system shows, on increasing pressure, four polymorphs with rock salt- (Fmm), KOH- (P2(1)/m), TlI- (Cmcm), and CsCl- (Pmm) type structures. The method of constant-pressure enthalpy minimization was used for all phases, by fully relaxing the corresponding crystal structures. Periodic ab initio energy calculations were performed by the CRYSTAL03 code, employing a DFT-GGA-PBE functional with a localized basis set of Gaussian-type functions. The three phase transitions were predicted to occur at 3.5, 6.0, and 17.7 GPa, respectively, against pressures of 6.6, 10.8, and 17 GPa from literature experimental results. The rock salt- to KOH-type and KOH- to TlI type displacive transformations show a weak first-order character. The TlI- to CsCl-type reconstructive transition is sharply first-order, and its kinetic mechanism was studied in detail on the basis of a P2(1)/m pathway, similar to that previously found for the rock salt- to CsCl-type transformation of NaCl. An activation enthalpy of 0.011 eV was found at the equilibrium pressure of 17.7 GPa. PMID- 16471719 TI - General, spontaneous ion replacement reaction for the synthesis of micro- and nanostructured metal oxides. AB - A novel spontaneous ion replacement route based on the solubility difference as the driving force to synthesize a number of metal oxides has been established. We present a comprehensive study on the ion replacement reaction for chemical synthesis of micro- and nanostructured Mn2O3, ZnO, CuO, CdO, Al2O3, and CaO samples. This novel approach described herein is derived from the solubility difference between two carbonate salts, in which a metal cation can be driven from one liquid phase into another solid phase in the solution system. The resulting metal carbonate salts are initially formed and subsequently calcined to form highly crystallined metal oxides. The variation of pH values, reaction temperature, and reagent shapes can vary the solubility of these two carbonate salts, which thus changes the final morphology of metal oxides. The present work makes a progress to simply and mildly synthesize metal oxides with various morphologies, due to the fact that materials with a desired morphology are a key engineering step toward their shape-dependent chemical and physical properties. PMID- 16471720 TI - Control of hole opening in single-wall carbon nanotubes and single-wall carbon nanohorns using oxygen. AB - Due to the simplicity of the process, holes in the graphene walls of single-wall carbon nanotubes (SWNTs) and single-wall carbon nanohorns (SWNHs) have often been opened using O2 gas at high temperatures, even though this contaminates the nanotubes with carbonaceous dust (C-dust). To open holes with less C-dust contamination, we found that a slow temperature increase of 1 degrees C/min or less, in air, was effective. We also found that SWNHs having little C-dust could store a large quantity of materials inside the tubes. We infer that the local temperature increase due to the exothermic reaction of combustion may have been suppressed in the slow combustion process, which was effective in reducing the C dust. PMID- 16471721 TI - Intracrystalline structure and physicochemical properties of mixed SiO2-TiO2 sol pillared aluminosilicate. AB - Highly porous layered inorganic-inorganic nanohybrids were prepared by pillaring SiO2-TiO2 nanosol particles with aluminosilicate layers. According to powder X ray diffraction analysis, the basal spacing of SiO2-TiO2 pillared aluminosilicate (STPC) calcined at 400 degrees C was determined to be larger than 40 A. N2 adsorption-desorption isotherm measurements showed the STPC to have a large Brunauer-Emmett-Teller surface area of approximately 590 m2/g, of which approximately 70% originates from micropores with a size range of 8-16 A. The sorption behavior of various solvent vapors such as hexane, methanol, and water reveals internal pore surfaces of the STPC to be hydrophobic. A distinct blue shift of absorption edge in UV-vis spectra clearly demonstrates that the nanosized TiO2 particles are formed between silicate layers as a pillar. Fourier transform infrared and extended X-ray absorption fine structure analysis at the Ti K edge reveals that the pillared titania exists in the form of anatase structured TiO2 nanocrystals, not in the form of covalently bonded mixed particles of TiO2-SiO2. On the basis of the present findings, we are able to conclude that the quantum-sized TiO2 and SiO2 particles are independently intercalated to form a multilayer stacking intracrystalline structure in the gallery space of aluminosilicate clay. PMID- 16471722 TI - Heterostructured nanohybrid of zinc oxide-montmorillonite clay. AB - We have synthesized heterostructured zinc oxide-aluminosilicate nanohybrids through a hydrothermal reaction between the colloidal suspension of exfoliated montmorillonite nanosheets and the sol solution of zinc acetate. According to X ray diffraction, N2 adsorption-desorption isotherm, and field emission-scanning electron microscopic analyses, it was found that the intercalation of zinc oxide nanoparticles expands the basal spacing of the host montmorillonite clay, and the crystallites of the nanohybrids are assembled to form a house-of-cards structure. From UV-vis spectroscopic investigation, it becomes certain that calcined nanohybrid contains two kinds of the zinc oxide species in the interlayer space of host lattice and in mesopores formed by the house-of-cards type stacking of the crystallites. Zn K-edge X-ray absorption near-edge structure/extended X-ray absorption fine structure analyses clearly demonstrate that guest species in the nanohybrids exist as nanocrystalline zinc oxides with wurzite-type structure. PMID- 16471723 TI - Guest-host colloid crystals: experimental study and simulations. AB - Disorder in colloid crystals was induced by doping them with a different number of large or small guest particles, which had a different deviation in size from the host colloids. The change in optical properties of the guest-host colloid crystals was assessed by using optical transmission spectroscopy while the variation in crystal structure was examined using scanning electron microscopy (SEM). The disruption in the crystalline lattices depended on the relative deviation in sizes of the guest and host particles and the concentration of the guest colloids. In parallel with experiments, the packing of spheres in guest host crystals was modeled with a simulated annealing algorithm. A good correlation was found between the changes in crystal structure observed by SEM imaging and the simulated sphere packing. The experimental and simulated changes in the transmission spectra of guest-host colloid crystals were in good agreement. PMID- 16471724 TI - Praseodymium hydroxide and oxide nanorods and Au/Pr6O11 nanorod catalysts for CO oxidation. AB - Praseodymium hydroxide nanorods were synthesized by a two-step approach: First, metallic praseodymium was used to form praseodymium chloride, which reacted subsequently with KOH solution to produce praseodymium hydroxide. In the second step the hydroxide was treated with a concentrated alkaline solution at 180 degrees C for 45 h, yielding nanorods as shown by the scanning and transmission electron microscopy images. The results of X-ray diffraction and energy dispersive X-ray spectroscopy experiments indicate that these nanorods are pure praseodymium hydroxide with a hexagonal structure, which can be converted into praseodymium oxide (Pr6O11) nanorods of a face-centered cubic structure after calcination at 600 degrees C for 2 h in air. Gold was loaded on the praseodymium oxide nanorods using HAuCl4 as the gold source, and NaBH4 was used to reduce the gold species to metallic nanoparticles with sizes of 8-12 nm on the nanorod surface. These Au/Pr6O11 nanorods exhibit superior catalytic activity for CO oxidation. PMID- 16471725 TI - Structural hierarchy in melt-processed poly(3-hexyl thiophene)-montmorillonite clay nanocomposites: novel physical, mechanical, optical, and conductivity properties. AB - Poly(3-hexyl thiophene) (P3HT) organically modified montmorillonite (om-MMT) polymer nanocomposites (PNCs) are prepared in the melt-cooled state. Hierarchical structures up to third order, namely, side chain mesomorph formation followed by the interchain lamellar structure of P3HT and finally its intercalation within the clay tactoids are observed. The structures are supported by transmission electron microscopy (TEM) and wide-angle X-ray scattering (WAXS) experiments. The TGA curves show two-stage degradation corresponding to those of the side chain and main chain of P3HT, and both temperatures decrease with an increase in clay concentration in the PNCs. The melting points of PNCs have increased by 2-3 degrees C higher than that of P3HT. The glass-transition temperature (Tg) and beta-transition temperature (Tbeta), measured by DMA, increase with an increase in clay concentration. The storage modulus (G) of PNCs has also increased more dramatically than that of P3HT. The UV-vis spectra of the PNCs show a blue shift in the pi-pi* absorption peak of the conjugated chain, but the photoluminescence spectra showed a red shift with an increase in the clay concentration. The quantum yield of the photoluminescence process also increases in the melt-cooled PNCs, and this is in sharp contrast to that of solvent cast PNCs where photoluminescence quenching was observed. Fibrillar network structure of the solvent cast PNCs promotes energy transfer of the charge carriers, but its absence in the melt-cooled films inhibits such energy transfer, increasing the quantum yield. The room-temperature dc conductivity of the PNCs decreased by an order compared to that of P3HT in both the doped and undoped states. The I-V characteristic curve shows semiconducting behavior, and it slowly transforms into insulator with increasing clay concentration. PMID- 16471726 TI - Well-aligned arrays of CuO nanoplatelets. AB - This paper reports well-aligned arrays of CuO nanoplatelets synthesized through a hydrothermal route without template's assistance. The surface of well-aligned arrays of CuO nanoplatelets looks like a wall. These nanoplatelets, possessing four clear edges, are 50-80 nm in thickness, 150-250 nm in width, and 0.8-1.5 microm in length. Electron microscopic detection shows that the nanoplatelet grows along the [010] direction. The Ostwald ripening mechanism has been used to describe the growth of CuO nanoplatelets. In addition, the optic and electrochemical properties of as-obtained products have been discussed. And the arrays of CuO nanoplatelets exhibit the blue shift in UV-visible spectra, a slow capacity fading rate, and a relatively high Coulombic efficiency in charge discharge process. PMID- 16471727 TI - Morphology of nanostructured platinum in mesoporous materials-effect of solvent and intrachannel surface. AB - An intrachannel surface of host silica was functionalized through the reaction of surface silanol groups with silanes to generate a monolayer of positively charged groups, and together with the strongly adsorbed and negatively charged PtCl6(2-), resulting in nanostructured platinum-mesoporous silica composites. The highly dispersed Pt nanoparticles and nanonetworks are fabricated from (CH3O)3Si(CH2)3N(CH3)3+Cl- functionalized mesoporous silica MCM-48 with H2PtCl6 in ethanol and water solvent, and characterized by PXRD, XAS, TEM, and N2 adsorption. The solvent of H2PtCl6 solution is found to affect the mobility of Pt precursors and the resulting morphology of nanostructured metallic Pt. The effect of the intrachannel surface properties on the incorporation and the morphology of nanostructured Pt on the deposition of Pt(NH3)4Cl2 and H2PtCl6 on Al-doped or C coated mesoporous silica MCM-41 is also studied relative to that on pure silica MCM-41. PMID- 16471728 TI - Solvent-induced novel morphologies in diblock copolymer blend thin films. AB - We report the morphology and phase behaviors of blend thin films containing two polystyrene-b-poly(methyl methacrylate) (PS-b-PMMA) diblock copolymers with different blending compositions induced by a selective solvent for the PMMA block, which were studied by transmission electron microscopy (TEM). The neat asymmetric PS-b-PMMA diblock copolymers employed in this study, respectively coded as a1 and a2, have similar molecular weights but different volume fractions of PS block (fPS=0.273 and 0.722). Another symmetric PS-b-PMMA diblock copolymer, coded as s, which has a PS block length similar to that of a1, was also used. For the asymmetric a1/a2 blend thin films, circular multilayered structures were formed. For the asymmetric a1/symmetric s blend thin films, inverted phases with PMMA as the dispersed domains were observed, when the weight fraction of s was less than 50%. The origins of the morphology formation in the blend thin films via solvent treatment are discussed. Combined with the theoretical prediction by Birshtein et al. (Polymer 1992, 33, 2750), we interpret the formation of these special microstructures as due to the packing frustration induced by the difference in block lengths and the preferential interactions between the solvent and PMMA block. Results obtained here suggest that diblock copolymer blend thin films treated with a selective solvent offer an alternative and attractive approach to control the self-organization of polymers. PMID- 16471730 TI - Two-dimensional crystal growth and stacking of bis(phthalocyaninato) rare earth sandwich complexes at the 1-phenyloctane/graphite interface. AB - Initial stages of two-dimensional crystal growth of the double-decker sandwich complex Lu(Pc*)2 [Pc* = 2,3,9,10,16,17,23,24-octakis(octyloxy)phthalocyaninato] have been studied by scanning tunneling microscopy at the liquid/solid interface between 1-phenyloctane and highly oriented pyrolytic graphite. High-resolution images strongly suggest alignment of the double-decker molecules into monolayers with the phthalocyanine rings parallel to the surface. Domains were observed with either hexagonal or quadrate packing motifs, and the growing interface of the layer was imaged. Molecular resolution was achieved, and the face of the phthalocyanine rings appeared as somewhat diffuse circular features. The alkyl chains are proposed to be interdigitating to maintain planar side-by-side packing. PMID- 16471729 TI - Catalyst-nanostructure interaction in the growth of 1-D ZnO nanostructures. AB - Vapor-liquid-solid is a well-established process in catalyst guided growth of 1-D nanostructures, i.e., nanobelts and nanowires. The catalyst particle is generally believed to be in the liquid state during growth, and is the site for impinging molecules. The crystalline structure of the catalyst may not have any influence on the structure of the grown nanostructures. In this work, using Au guided growth of ZnO, we show that the interfaces between the catalyst droplet and the nanostructure grow in well-defined mutual crystallographic relationships. The nanostructure defines the crystallographic orientation of the solidifying Au droplet. Possible alloy, intermetallic, or eutectic phase formation during catalysis are elucidated with the help of a proposed ternary Au-Zn-O phase diagram. PMID- 16471731 TI - Grafting and polymer formation on silicon from unsaturated Grignards: I-aromatic precursors. AB - Anodic decomposition of a phenylmagnesium halide at a surface-hydrogenated silicon electrode leads to formation of polymeric layers covalently anchored to the silicon surface. These layers have been characterized using spectroellipsometry, photoluminescence, infrared, and X-ray photoelectron spectroscopies. The phenyl ring appears preserved in the process, and the polymer formed is a polyphenylene. Contamination by aliphatic groups from the solvent may be minimized by using a solvent resistant to hydrogen abstraction by the phenyl radicals. Regioselectivity of the branching may be oriented to the para form by using 4-chlorophenylmagnesium bromide as the precursor. PMID- 16471732 TI - Infrared reflection-absorption spectra of C2H4 and C2H6 on Cu: effect of surface roughness. AB - Infrared reflection absorption spectroscopy (IRRAS) of the highly symmetric molecules C2H4 and C2H6 adsorbed as mono- and multilayers onto copper films is studied in relation to the type of metal-film roughness. Spectra of C2H4 show Raman lines on cold-deposited Cu films but not on Cu deposited at room temperature. For C2H6, the IR spectra from both types of metal films are similar; the surface infrared selection rule holds and no Raman bands are observed. The Raman lines that appear in the IR spectra already at low exposures are attributed to species adsorbed at special defect sites, identical to the so-called active sites in surface enhanced Raman scattering (SERS). The IR excitation mechanism by transient electron transfer to the adsorbate pi* state can deliver a discrete vibrational band of a Raman-active vibration only under certain circumstances, for example, for adsorbates at the "SERS-active sites". C2H6 at these sites cannot deliver Raman bands in IRRAS, because it has no pi* state. We also discuss IRRAS measurements on Cu(111) and Cu(110) single crystals, where Raman bands of C2H4 have been observed. PMID- 16471733 TI - Characterization of the local structures of Ti-MCM-41 and their photocatalytic reactivity for the decomposition of NO into N2 and O2. AB - Ti-MCM-41 mesoporous molecular sieves were prepared at ambient temperature and were characterized by X-ray absorption near-edge structure and extended X-ray absorption fine structure, UV-vis, Fourier transform infrared spectroscopy, and photoluminescence spectroscopic analyses. It was found that an increase in the Ti content caused the structure of the Ti-oxides in Ti-MCM-41 to change from an isolated tetrahedral coordination to adjacent Ti-oxide species with Ti4+ of tetrahedral coordination. The photocatalytic reactivity of these catalysts for the decomposition of NO into N2 and O2 was found to strongly depend on the local structure of the Ti-oxide species including their coordination and distribution, i.e., the charge transfer excited state of the highly dispersed isolated tetrahedrally coordinated Ti-oxides act as the active sites for the photocatalytic decomposition of NO into N2 and O2. PMID- 16471734 TI - Reforming of oxygenates for H2 production: correlating reactivity of ethylene glycol and ethanol on Pt(111) and Ni/Pt(111) with surface d-band center. AB - The dehydrogenation and decarbonylation of ethylene glycol and ethanol were studied using temperature programmed desorption (TPD) on Pt(111) and Ni/Pt(111) bimetallic surfaces, as probe reactions for the reforming of oxygenates for the production of H2 for fuel cells. Ethylene glycol reacted via dehydrogenation to form CO and H2, corresponding to the desired reforming reaction, and via total decomposition to produce C(ad), O(ad), and H2. Ethanol reacted by three reaction pathways, dehydrogenation, decarbonylation, and total decomposition, producing CO, H2, CH4, C(ad), and O(ad). Surfaces prepared by deposition of a monolayer of Ni on Pt(111) at 300 K, designated Ni-Pt-Pt(111), displayed increased reforming activity compared to Pt(111), subsurface monolayer Pt-Ni-Pt(111), and thick Ni/Pt(111). Reforming activity was correlated with the d-band center of the surfaces and displayed a linear trend for both ethylene glycol and ethanol, with activity increasing as the surface d-band center moved closer to the Fermi level. This trend was opposite to that previously observed for hydrogenation reactions, where increased activity occurred on subsurface monolayers as the d-band center shifted away from the Fermi level. Extrapolation of the correlation between activity and the surface d-band center of bimetallic systems may provide useful predictions for the selection and rational design of bimetallic catalysts for the reforming of oxygenates. PMID- 16471735 TI - Structure and desorption energetics of ultrathin D2O ice overlayers on serine- and serinephosphate-terminated self-assembled monolayers. AB - This paper reports on the structure and desorption dynamics of thin D2O ice overlayers (0.2-10 monolayers) deposited on serine- and serinephosphate- (with H+, Na+, Ca2+ counterions) terminated self-assembled monolayers (SAMs). The D2O ice overlayers are deposited on the SAMs at approximately 85 K in ultrahigh vacuum and characterized with infrared reflection absorption spectroscopy (IRAS). Reflection absorption (RA) spectra obtained at sub-monolayer D2O coverage reveal that surface modes, e.g. free dangling OD stretch, dominate on the serine SAM surface, whereas vibrational modes characteristic for bulk ice are more prominent on the serinephosphate SAMs. Temperature programmed desorption mass spectrometry (TPD-MS) and TPD-IRAS are subsequently used to investigate the energetics and the structural transitions occurring in the ice overlayer during temperature ramping. D2O ice (approximately 2.5 monolayers) on the serine SAMs undergoes a gradual change from an amorphous- to a crystalline-like phase upon increasing the substrate temperature. This transition is not as pronounced on the serine phosphate SAM most likely because of reduced mobility due to strong pinning to the surface. We show also that the energy of desorption for a sub-monolayer of D2O ice on serinephosphate SAM surfaces with a Na+ and Ca2+ counterions is equally high or even exceeds previously reported values for analogous high-energy SAMs. PMID- 16471736 TI - Electronic excited states of Si(100) and organic molecules adsorbed on Si(100). AB - The electronically excited states of the Si(100) surface and acetylene, benzene, and 9,10-phenanthrenequinone adsorbed on Si(100) are studied with time-dependent density functional theory. The computational cost of these calculations can be reduced through truncation of the single excitation space. This allows larger cluster models of the surface in conjunction with large adsorbates to be studied. On clean Si(100), the low-lying excitations correspond to transitions between the pi orbitals of the silicon-silicon dimers. These excitations are predicted to occur in the range 0.4-2 eV. When organic molecules are adsorbed on the surface, surface --> molecule, molecule --> surface, and electronic excitations localized within the adsorbate are also observed at higher energies. For acetylene and benzene, the remaining pipi* excitations are found to lie at lower energies than in the corresponding gas-phase species. Even though the aromaticity of 9,10 phenanthrenequinone is retained, significant shifts in the pipi* excitations of the aromatic rings are predicted. This is in part due to structural changes that occur upon adsorption. PMID- 16471737 TI - Hexane isomerization and cracking activity and intrinsic acidity of H-zeolites and sulfated zirconia-titania. AB - Adsorption of N2 was studied on zeolite H-Y, ultrastabilized H-Y (H-USY), H mordenite, H-ZSM-5, H-beta, and on sulfated zirconia-titania (SZT) mixed oxide by diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) at 298 K and at N2 pressures up to 9 bar. The adsorption-induced DeltanuOH red-shift of the nuOH bands was used as a measure of the intrinsic acid strength of the Brnonsted acid sites. The intrinsic acid strength of the solids follows the order of H-ZSM 5 approximately H-mordenite approximately H-beta > H-USY > SZT approximately H-Y. The solids were characterized by their hexane conversion activities at 553 K and 6.1 kPa hexane partial pressure. The reaction was shown to proceed predominantly by a bimolecular mechanism, while the reaction was first order in hexane and zero order in alkenes. The site-specific apparent rate constant of the bimolecular hexane conversion was shown to parallel the intrinsic acid strength of the samples, suggesting that the ratio of the apparent and the intrinsic activity, that is, the KA' equilibrium constant of alkane adsorption on the hydrocarbon covered sorption sites, is hardly dependent on the catalyst structure. PMID- 16471738 TI - Characterization of Langmuir-Blodgett films of a ferroelectric liquid crystal. AB - Molecular orientation, structure, and phase transition behaviors in Langmuir Blodgett (LB) and cast films of a ferroelectric liquid crystal of sec-butyl 6-(4 (nonyloxy)benzoyloxy)-2-naphthoate (FLC-1) are determined by ultraviolet (UV) spectroscopy, X-ray diffraction, and infrared (IR) spectroscopy. It is found that the orientation angle of chromophores theta in LB films is 41 degrees from the surface normal. The tilt angle of the chromophore changes at 56, 70, and 88 degrees C, respectively, which denotes the presence of phase transitions. Two kinds of layered or isomeric crystal structures of the LB films with layer spacings of 3 and 3.5 nm at room temperature have been found while the latter disappears above 45 degrees C, as confirmed by measurement of temperature dependent IR spectra. PMID- 16471739 TI - Absolute orientation of molecules at interfaces. AB - A method to determine the absolute orientation of molecules at liquid interfaces by sum frequency generation (SFG) is reported. It is based on measurements of the orientations of two nonparallel vibrationally active chromophores in the molecule of interest combined with a rotation matrix formulation to obtain the absolute molecular orientation. We chose m-tolunitrile, a planar molecule adsorbed to the air/water interface, as a proof-of-method experiment. Quantitative analysis of different polarization sum frequency intensities facilitate unique peak assignments of the methyl and nitrile groups of m-tolunitrile. The SFG analysis of the measurement yields a nitrile group tilting at 53 degrees to the surface normal, and the C3 axis of the methyl group is almost upright at 23 degrees with respect to the surface normal. Using a rotation matrix formulation, we found that the angle between the surface plane and the m-tolunitrile molecular plane is 70 degrees. PMID- 16471740 TI - Quantum sieving in single-walled carbon nanotubes: effect of interaction potential and rotational-translational coupling. AB - The selective adsorption of heavy isotopes in narrow nanotubes, known as quantum sieving, is studied using a simple approximate theory for several different potential models. We address the reasons for wide disagreement among previously published results for quantum sieving. We analyze the sensitivity of quantum sieving to perturbations in the potential parameters used in the calculations. The selectivities are very sensitive to changes in the atomic diameter parameter and less sensitive to changes in the potential well depth. We present an approximate method for accounting for rotational-translational coupling that is computationally efficient and accurate for the narrowest nanotubes. For wide nanotubes, the estimation of rotational-translational coupling becomes inaccurate because of neglect of the effect of rotational states on the translational degrees of freedom. PMID- 16471741 TI - Quantum States of hydrogen and its isotopes confined in single-walled carbon nanotubes: dependence on interaction potential and extreme two-dimensional confinement. AB - Quantum mechanical energy levels are computed for the hydrogen molecule and its homonuclear isotopes confined within carbon nanotubes of various sizes and structures using three different interaction potentials. Two translational and two rotational degrees of freedom are treated explicitly. We study the dependence on the interaction potential and the size of the nanotube of several features, including zero-pressure quantum sieving selectivities, ortho-para energy splittings, and wave function characteristics. We show that large quantum sieving selectivities, as well as large deviations from gas phase ortho-para splittings, occur only under the condition of extreme two-dimensional confinement, when the characteristic length of the hydrogen-carbon interaction potential is nearly equal to the radius of the nanotube. PMID- 16471742 TI - Surface immobilization and mechanical properties of catanionic hollow faceted polyhedrons. AB - We report here for the first time on surface immobilization of hollow faceted polyhedrons formed from catanionic surfactant mixtures. We find that electrostatic interaction with the substrate dominates their adhesion behavior. Using polyelectrolyte coated surfaces with tailored charge densities, polyhedrons can thus be immobilized without complete spreading, which allows for further study of their mechanical properties using AFM force measurements. The elastic response of individual polyhedrons can be locally resolved, showing pronounced differences in stiffness between faces and vertexes of the structure, which makes these systems interesting as models for structurally similar colloidal scale objects such as viruses, where such effects are predicted but cannot be directly observed due to the smaller dimensions. Elastic constants of the wall material are estimated using shell and plate deformation models and are found to be a factor of 5 larger than those for neutral lipidic bilayers in the gel state. We discuss the molecular origins of this high stiffness. PMID- 16471743 TI - Influence of the hydroxylation of gamma-Al2O3 surfaces on the stability and diffusion of single Pd atoms: a DFT study. AB - Using recent well-defined models of gamma-Al2O3 surfaces, we study the interaction of single Pd atoms with gamma-Al2O3 surfaces corresponding to realistic pretreatment conditions by means of density functional theory periodic calculations. For relevant hydroxylation states of the surface, we determine potential energy surfaces (PES) that depict the relationship between structure and interaction at the metal-oxide interface. This approach enables the determination of the low-energy diffusion paths of the adsorbed Pd species. We applied classical transition-state theory to derive the temperature-dependent hopping rate of Pd on gamma-Al2O3 surfaces. Our work provides new insight into the chemisorption and diffusion process of single Pd atoms on alumina and show that the binding energy and hopping rate of Pd atoms decrease as the surface OH coverage increases. These results offer new highlights on Pd cluster formation at the initial nucleation steps on gamma-Al2O3 surfaces. PMID- 16471744 TI - Electrochemistry of nanopore electrodes in low ionic strength solutions. AB - The steady-state voltammetric behavior of truncated conical nanopore electrodes (20-200 nm orifice radii) has been investigated in low ionic strength solutions. Voltammetric currents at the nanopore electrode reflect both diffusive and migrational fluxes of the redox molecule and, thus, are strongly dependent on the charge of the redox molecule and the relative concentrations of the supporting electrolyte and redox molecule. In acetonitrile solutions, the limiting current for the oxidation of the positively charged ferrocenylmethyltrimethylammonium ion is suppressed at low supporting electrolyte concentrations, while the limiting current for the oxidation of the neutral species ferrocene is unaffected by the ionic strength. The dependence of the limiting current on the relative concentrations of the supporting electrolyte and redox molecule is accurately predicted by theory previously developed for microdisk electrodes. Anomalous values of the voltammetric half-wave potential observed at very small nanopore electrodes (<50 nm radius orifice radii) are ascribed to a boundary potential between the pore interior and bulk solution (i.e., a Donnan-type potential). PMID- 16471745 TI - Catalysis by oxide-supported clusters of iridium and rhodium: hydrogenation of ethene, propene, and toluene. AB - The hydrogenation reactions of ethene, propene, and toluene were used as probes of the catalytic properties of small clusters of rhodium (Rh6) and of iridium (Ir4 and Ir6) (as well as of larger aggregates of these metals) on oxide supports (gamma-Al2O3, MgO, and La2O3). The catalysts were characterized in the working state by extended X-ray absorption fine structure (EXAFS) spectroscopy, providing evidence of the cluster structures and cluster-support interactions; by infrared spectroscopy, providing evidence of hydrocarbon adsorbates and possible reaction intermediates on the clusters; and by kinetics of the hydrogenation reactions. The EXAFS data indicate that the metal clusters, while remaining intact and maintaining their bonding to the support during catalysis, underwent slight rearrangements to accommodate reactive intermediates. As the concentrations of reactive intermediates such as pi-bonded alkenes and alkyls on the clusters increased, the cluster frames swelled, and the clusters flexed away from the support. The data indicate self-inhibition of reaction by adsorbed hydrocarbons and differences between ethene hydrogenation and propene hydrogenation that may arise primarily from different adsorbate-adsorbate interactions. PMID- 16471746 TI - O2 reduction on graphite and nitrogen-doped graphite: experiment and theory. AB - An experimental and theoretical study of electroreduction of oxygen to hydrogen peroxide is presented. The experimental measurements of nitrided Ketjenblack indicated an onset potential for reduction of approximately 0.5 V (SHE) compared to the onset potential of 0.2 V observed for untreated carbon. Quantum calculations on cluster models of nitrided and un-nitrided graphite sheets show that carbon radical sites formed adjacent to substitutional N in graphite are active for O2 electroreduction to H2O2 via and adsorbed OOH intermediate. The weak catalytic effect of untreated carbon is attributed to weaker bonding of OOH to the H atom-terminated graphite edges. Substitutional N atoms that are far from graphite sheet edges will be active, and those that are close to the edges will be less active. Interference from electrochemical reduction of H atoms on the reactive sites is considered, and it is shown that in the potential range of H2O2 formation the reactive sites are not blocked by adsorbed H atoms. PMID- 16471747 TI - Time-dependent organization and wettability of decanethiol self-assembled monolayer on Au(111) investigated with STM. AB - A detailed study on the time-dependent organization of a decanethiol self assembled monolayer (SAM) at a designed solution concentration onto a Au(111) surface has been performed with scanning tunneling microscopy (STM). The SAMs were prepared by immersing Au(111) into an ethanol solution containing 1 microM decanethiol with different immersion times. STM images revealed the formation process and adlayer structure of the SAMs. It was found that the molecules self organized into adlayers from random separation to a well-defined structure. From 10 s, small domains with ordered molecular organization appeared, although random molecules could be observed on Au(111) at the very initial stage. At 30 s, the SAM consisted of uniform short stripes. Each stripe consisted of sets of decanethiol mainly containing eight molecules. With the immersion time increasing, the length of the stripes increased. At 5 min, the alkyl chains overlapped each other between the adjacent stripes, indicating the start of a stacked process. After immersing Au(111) in decanethiol solution for 3 days, a densely packed adlayer with a (radical 3 x radical 3)R30 degrees structure was observed. The formation process and structure of decanethiol SAMs are well related to sample preparation conditions. The wettability of the decanethiolate SAM-modified Au(111) surface was also investigated. PMID- 16471748 TI - Structure of the liquid-vacuum interface of room-temperature ionic liquids: a molecular dynamics study. AB - Molecular dynamics simulations for the liquid-vacuum interface of the ionic liquid 1-ethyl-3-methylimidazolium nitrate (EMIM+/NO3-) were performed for both electronically polarizable and nonpolarizable potential energy surfaces. The interfacial structural properties, such as the oscillation in the number density profile, the orientational ordering, and the local clustering in the interfacial region, were calculated. The simulations with both the polarizable and nonpolarizable model demonstrate the existence of an inhomogeneous interfacial structure normal to the surface layer. It was found for both models that the ethyl tail group on EMIM+ is likely to protrude outward from the surface. In the outmost surface layer, the cation is likely to lie on the surface with the imidazolium ring parallel to the interface, while there is a second region with enhanced density from that in the bulk where the cation preferably slants with the imidazolium ring tending to be perpendicular to the surface. The results also reveal that the electronic polarization effect is important for the ionic liquid interface. It is found that the cation is likely to be segregated at the ionic liquid surface for the polarizable model, while for the nonpolarizable model, the anion is found to be more likely to exhibit such behavior. The surface tension of the polarizable model (58.5 +/- 0.5 mN/m) is much smaller than that of the nonpolarizable model (82.7 +/- 0.6 mN/m), in better agreement with extrapolated experimental measurements on similar ionic liquid systems. PMID- 16471749 TI - Sum frequency generation microscopy of microcontact-printed mixed self-assembled monolayers. AB - Sum frequency generation imaging microscopy (SFGIM) is used to image the chemically distinct regions of a microcontact-printed monolayer surface. The contrast in the images is based on the vibrational spectrum of each component in the monolayer. Mixtures of C16 thiols on gold with CH3 and phenyl termination are imaged with a resolution of approximately 10 microm. Microcontact printing produces films that are different compared to the immersion procedure of forming self-assembled monolayers. The SFGIM technique is able to obtain a vibrational spectrum at each point on the surface and demonstrate that the stamped area has significant mixing with the molecules deposited from the backfilling solution. PMID- 16471750 TI - Catalytic effects of subsurface carbon in the chemisorption of hydrogen on a Mg(0001) surface: an ab-initio study. AB - Ab initio density functional theory (DFT) calculations are performed to explore possible catalytic effects on the dissociative chemisorption of hydrogen on a Mg(0001) surface when carbon is incorporated into Mg materials. The computational results imply that a C atom located initially on a Mg(0001) surface can migrate into the subsurface and occupy an fcc interstitial site, with charge transfer to the C atom from neighboring Mg atoms. The effect of subsurface C on the dissociation of H2 on the Mg(0001) surface is found to be relatively marginal: a perfect sublayer of interstitial C is calculated to lower the barrier by 0.16 eV compared with that on a pure Mg(0001) surface. Further calculations reveal, however, that sublayer C may have a significant effect in enhancing the diffusion of atomic hydrogen into the sublayers through fcc channels. This contributes new physical understanding toward rationalizing the experimentally observed improvement in absorption kinetics of H2 when graphite or single walled carbon nanotubes (SWCNT) are introduced into the Mg powder during ball milling. PMID- 16471751 TI - TOF-SIMS evidence of intercalated molecular gases and diffusion-limited reaction kinetics in an alpha particle-irradiated PTFE matrix. AB - The chemical evolution of poly(tetrafluoroethylene) (PTFE) that is brought about by increasing levels of irradiation with alpha particles is accompanied by the emergence and proliferation of functionalized moieties. Families of reaction products specifically identified in the alpha-irradiated polymer matrix include hydride-, hydroxide-, and oxide-functionalized fluorocarbons. The data also indicate the emergence of hydrogen peroxide (H2O2) and hydrazine (N2H4), but no distinct evidence suggesting the formation of perfluorinated amines, amides, or cyanogens is found. In this article we substantiate the speciation of emergent species and reveal evidence of intercalated molecular gases with which alpha particle-generated radicals may react to form the observed products. Furthermore, we present evidence to suggest that the kinetics of alpha particle-induced reaction is limited by the diffusion of radicals within the polymer matrix. That is to say, chemical additives in the polymer matrix are shown to be scavengers of H*, O*, and F* radicals and limit the rates of reaction that produce functionalized fluorocarbon moieties. Above a threshold dose of alpha particles, the concentration of radicals exceeds that of the scavenger species, and free radical diffusion commences as evidenced by a sudden increase in the yield of reaction products. Samples of PTFE were irradiated to alpha doses in the range of 10(7) to 5 x 10(10) rad with 5.5 MeV 4He2+ ions from a tandem accelerator. Residual gas analysis (RGA) was utilized to monitor the liberation of molecular gases from PTFE during alpha particle irradiation of samples in vacuum. Static time-of-flight SIMS (TOF-SIMS), equipped with a 20 keV C60+ source, was employed to probe chemical changes as a function of alpha particle irradiation. Chemical images and high-resolution mass spectra were collected in both the positive and negative polarities. PMID- 16471752 TI - Ice nucleation and phase behavior on oligo(ethylene glycol) and hydroxyl self assembled monolayers: simulations and experiments. AB - The nucleation and phase behavior of ultrathin D2O-ice overlayers have been studied on oligo(ethylene glycol) (OEG)-terminated and hydroxyl self-assembled monolayers (SAMs) at low temperatures in ultrahigh vacuum. Infrared reflection absorption spectroscopy (IRAS) is used to characterize the ice overlayers, the SAMs, and the interactions occurring between the ice and the SAM surfaces. Spectral simulations, based on optical models in conjunction with Maxwell Garnett effective medium theory, point out the importance of including voids in the modeling of the ice structures, with void fractions reaching 60% in some overlayers. The kinetics of the phase transition from amorphous-like to crystalline-like ice upon isothermal annealing at 140 K is found to depend on the conformational state of the supporting OEG SAM surface. The rate is fast on the helical OEG SAMs and slow on the corresponding all-trans SAMs. This difference in kinetics is most likely due to a pronounced D2O interpenetration and binding to the all-trans segments of the ethylene glycol portion of the SAM. No such penetration and binding was observed on the helical OEG SAM. PMID- 16471753 TI - Adsorption characteristics of arylisocyanide on Au and Pt electrode surfaces: surface-enhanced Raman scattering study. AB - It is very important to understand metal-molecule interface characteristics for the development of efficient molecular wires in molecular electronics. Because isocyanide is potentially a good alligator clip, we have investigated in this work the adsorption characteristics of 2,6-dimethylphenylisocyanide (DMPI) on Au and Pt electrodes by recording the potential-dependent surface-enhanced Raman scattering (SERS) spectra. First of all, we confirmed that Pt nanoaggregate films were efficient SERS-active substrates, not only in ambient conditions, but also in electrochemical environments. Second, we confirmed that aryl isocyanide should adsorb on Au and Pt by forming exclusively metal-CN bonds, via a pure sigma type interaction in the case of gold compared with a sigma/pi synergistic interaction on Pt. This implies that DMPI should adsorb on Au only via the on-top site, whereas not only the on-top site, but also the 2-fold bridge and 3-fold hollow sites, could be used in the surface adsorption of DMPI on Pt. Despite these differences, DMPI was assumed to possess a vertical orientation with respect to the Au and Pt substrates, irrespective of the potential variation between +0.2 and -0.6 V relative to the Ag/AgCl reference electrode. The latter characteristics of the Au-CN and Pt-CN combinations are presumed to be useful in designing molecular-scale wires. PMID- 16471755 TI - Characteristic motion of a camphanic acid disk on water depending on the concentration of Triton X-100. AB - As a simple autonomous motor, the self-motion of a camphanic acid disk on the aqueous phase with a neutral surfactant (Triton X-100) was investigated. Whereas only continuous motion was observed on water, intermittent motion (alternating between motion and rest) was observed upon addition of Triton X-100. Under the experimental conditions that gave intermittent motion, the surface tension of the aqueous phase changed periodically, synchronous with the contact angle around the camphanic acid disk. These characteristics of self-motion are discussed in relation to the surface tension depending on the concentration of camphanic acid with or without Triton X-100 as the driving force of the motion. PMID- 16471754 TI - Distributions of ionic concentrations and electric field around the three-phase contact at high rates of Langmuir-Blodgett deposition. AB - A mathematical problem is formulated and numerically solved for addressing the electric field and ionic concentration distributions developing around the three phase contact line during the Langmuir-Blodgett deposition of charged monolayers. Compared to a previous paper dealing with the same effect (J. Phys. Chem. B 2004, 108, 13449), the present analysis is not restricted to the case of low deposition rates and small concentration changes. The obtained results show that, for sufficiently high deposition rates, the subphase composition substantially changes in the immediate vicinity of the three-phase contact line. It is shown that the predicted changes in the subphase composition can drastically affect the adhesion work and the dynamic contact angle. On this basis, the influence of the concentration polarization effect on meniscus behavior is discussed. PMID- 16471756 TI - In situ chemical reductive growth of platinum nanoparticles on indium tin oxide surfaces and their electrochemical applications. AB - Platinum nanoparticles directly attached to indium tin oxide (PtNP/ITO) were successfully fabricated by using an in situ chemical reductive growth method. In this method, PtNPs could be grown on the ITO surface via the one-step immersion into the growth solution containing PtCl4(2-) and ascorbic acid. The attached and grown PtNPs were spherical having an agglomerated nanostructure composed of small nanoclusters. From the morphological changes depending on the growth time, which were observed with an FE-SEM, PtNPs were found to be grown via the progressive nucleation mechanism. As the characteristics of the PtNP/ITO were those of a working electrode, it was found that the charge transfer resisivity was significantly lowered due to the grown PtNPs. Hence, for a typical redox system of [Fe(CN)6]3-/[Fe(CN)6]4-, the PtNP/ITO electrode exhibited the electrochemical responses similar to those of the bulk Pt electrode. Furthermore, it was clarified that the PtNP/ITO electrode had significant electrocatalytic properties for the oxygen reduction and methanol oxidation. The present PtNPs that had the agglomerated nanostructure may be promising for a new type of electrode material. PMID- 16471757 TI - Reorganization energy of electron transfer in viscous solvents above the glass transition. AB - We present a molecular-dynamics study of the solvent reorganization energy of electron transfer in supercooled water. We observe a sharp decrease of the reorganization energy at a temperature identified as the temperature of structural arrest due to cage effect as discussed by the mode coupling theory. Both the heat capacity and dielectric susceptibility of the pure water show sharp drops at about the same temperature. This temperature also marks the onset of the enhancement of translational diffusion relative to rotational relaxation signaling the breakdown of the Stokes-Einstein relation. The change in the reorganization energy at the transition temperature reflects the dynamical arrest of the slow, collective relaxation of the solvent related to Debye relaxation of the solvent dipolar polarization. PMID- 16471758 TI - Solvent size effect on the depletion layer of a polymer solution near an interface. AB - By varying polymer concentration phi0p and Flory-Huggins parameter chi, the effect of solvent size on the depletion interaction between polymer coils and a hard wall was investigated by the real-space version of self-consistent field theory (SCFT). The depletion profiles and depletion thickness indicated that the depletion effect is strong in less good solvent with large molecular volume. Through the analysis of the respective free energies of polymer coils and solvent molecules, we found that the increment in the translation entropy of the solvent is the key to strengthening the depletion interaction. On the basis of the SCFT results, we define a solvent with volume about one to six times that of the polymer segment as a "middle-sized solvent". The density oscillations previously studied by Van der Gucht et al. and Maassen et al. were also observed in our simulation, and the addition of middle-sized solvent will magnify the amplitude of the oscillations. The solvent-size-dependent depletion interaction may be an explanation for the reduced entanglement and promoted crystallization behavior of polymer coils prepared from the solution with middle-sized solvent. PMID- 16471759 TI - Unusual solvent effect on a SN2 reaction. A quantum-mechanical and kinetic study of the Menshutkin reaction between 2-amino-1-methylbenzimidazole and iodomethane in the gas phase and in acetonitrile. AB - The quaternization reaction between 2-amino-1-methylbenzimidazole and iodomethane was investigated in the gas phase and in liquid acetonitrile. Both experimental and theoretical techniques were used in this study. In the experimental part of this work, accurate second-order rate constants were obtained for this reaction in acetonitrile from conductivity data in the 293-323 K temperature range and at ambient pressure. From two different empirical equations describing the effect of temperature on reaction rates, thermodynamic functions of activation were calculated. In the theoretical part of this work, the mechanism of this reaction was investigated in the gas phase and in acetonitrile. Two different quantum levels (B3LYP/[6-311++G(3df,3pd)/LanL2DZ]//B3LYP/[6-31G(d)/LanL2DZ] and B3LYP/[6 311++G(3df,3pd)/LanL2DZ]//B3LYP/[6-31+G(d)/LanL2DZ]) were used in the calculations, and the acetonitrile environment was modeled using the polarized continuum model (PCM). In addition, an atoms in molecules (AIM) analysis was made aiming to characterize possible hydrogen bonding. The results obtained by both techniques are in excellent agreement and lead to new insight into the mechanism of the reaction under examination. These include the identification and thermodynamic characterization of the relevant stationary species, the rationalization of the mechanistic role played by the solvent and the amine group adjacent to the nucleophile nitrogen atom, the proposal of alternative paths on the modeled potential energy surfaces, and the origin of the marked non-Arrhenius behavior of the kinetic data in solvent acetonitrile. In particular, the AIM analysis confirmed the operation of intermolecular hydrogen bonds between reactants and between products, both in the gas phase and in solution. It is also concluded that the unusual solvent effect on this Menshutkin reaction stems from the conjunction of a nucleophile possessing a relatively complex chemical structure with a dipolar aprotic solvent that is protophobic. PMID- 16471760 TI - Factors governing the metal coordination number in metal complexes from Cambridge Structural Database analyses. AB - The metal coordination number (CN) is a key determinant of the structure and properties of metal complexes. It also plays an important role in metal selectivity in certain metalloproteins. Despite its central role, the preferred CN for several metal cations remains ambiguous, and the factors determining the metal CN are not fully understood. Here, we evaluate how the CN depends on (1) the metal's size, charge, and charge-accepting ability for a given set of ligands, and (2) the ligand's size, charge, charge-donating ability, and denticity for a given metal by analyzing the Cambridge Structural Database (CSD) structures of metal ions in the periodic table. The results show that for a given ligand type, the metal's size seems to affect its CN more than its charge, especially if the ligand is neutral, whereas, for a given metal type, the ligand's charge and charge-donating ability appear to affect the metal CN more than the ligand's size. Interestingly, all 98 metal cations surveyed could adopt more than than one CN, and most of them show an apparent preference toward even rather than odd CNs. Furthermore, as compared to the preferred metal CNs observed in the CSD, those in protein binding sites generally remain the same. This implies that the protein matrix (excluding amino acid residues in the metal's first and second coordination shell) does not impose severe geometrical restrictions on the bound metal cation. PMID- 16471762 TI - Effect of surface packing density of interfacially adsorbed monoclonal antibody on the binding of hormonal antigen human chorionic gonadotrophin. AB - Interfacial adsorption of a mouse monoclonal antibody (type IgG1, anti-beta-hCG) at the hydrophilic silicon oxide/water interface has been studied by spectroscopic ellipsometry and neutron reflection, followed by assessment of binding of a hormonal antigen, human chorionic gonadotrophin (hCG), onto the adsorbed antibody molecules. The amount of adsorption reached a maximum around the isoelectric pH (IP) of 6 for the antibody; this pH-dependent pattern could be altered by increasing salt concentration, a trend also observed for other proteins. Neutron reflection revealed the formation of a 40 A uniform layer from the adsorbed antibody, indicating a flat-on orientation. The subsequent hCG binding showed that the molar ratio of hCG bound to antibody at the interface was as high as 0.7 at low surface coverage of antibody and decreased with increasing surface antibody concentration. The results point to an increasing extent of steric hindrance to hCG access with increasing packing density of antibody molecules on the surface. Comparison with previously published crystal structure studies suggests twisting of the variable region to allow access of the antigen. The binding of hCG was also found to be pH-dependent with its maximum around the IP, if the ionic strength of the solution was low (20 mM). However, if the ionic strength was increased to 200 mM, then hCG binding was influenced by a combination of steric hindrance and electrostatic interaction between the antigen and the surface. These results are highly relevant to the improvement of the performance of biotechnologies such as fertility test pads and biosensors based on antibody immobilization. PMID- 16471761 TI - Fluorescence correlation spectroscopy simulations of photophysical phenomena and molecular interactions: a molecular dynamics/monte carlo approach. AB - Fluorescence correlation spectroscopy (FCS) is being applied increasingly to study diffusion and interactions of fluorescently labeled macromolecules in complex biological systems. Fluctuations in detected fluorescence, deltaF(t), are expressed as time-correlation functions, G(tau), and photon-count histograms, P(k;DeltaT). Here, we developed a generalized simulation approach to compute G(tau) and P(k;DeltaT) for complex systems with arbitrary geometry, photophysics, diffusion, and macromolecular interactions. G(tau) and P(k;DeltaT) were computed from deltaF(t) generated by a Brownian dynamics simulation of single-molecule trajectories followed by a Monte Carlo simulation of fluorophore excitation and detection statistics. Simulations were validated by comparing analytical and simulated G(tau) and P(k;DeltaT) for diffusion of noninteracting fluorophores in a three-dimensional Gaussian excitation and detection volume. Inclusion of photobleaching and triplet-state relaxation produced significant changes in G(tau) and P(k;DeltaT). Simulations of macromolecular interactions and complex diffusion were done, including transient fluorophore binding to an immobile matrix, cross-correlation analysis of interacting fluorophores, and anomalous sub and superdiffusion. The computational method developed here is generally applicable for simulating FCS measurements on systems complicated by fluorophore interactions or molecular crowding, and experimental protocols for which G(tau) and P(k;DeltaT) cannot be computed analytically. PMID- 16471763 TI - Conformational preference and cis-trans isomerization of 4(R)-substituted proline residues. AB - We report here the conformational preference and prolyl cis-trans isomerization of 4(R)-substituted proline dipeptides, N-acetyl-N'-methylamides of 4(R)-hydroxy L-proline and 4(R)-fluoro-L-proline (Ac-Hyp-NHMe and Ac-Flp-NHMe, respectively), studied at the HF/6-31+G(d), B3LYP/6-31+G(d), and B3LYP/6-311++G(d,p) levels of theory. The 4(R)-substitution by electron-withdrawing groups did not result in significant changes in backbone torsion angles as well as endocyclic torsion angles of the prolyl ring. However, the small changes in backbone torsion angles phi and psi and the decrease of bond lengths r(Cbeta-Cgamma) or r(Cgamma-Cdelta) appear to induce the increase of the relative stability of the trans up-puckered conformation and to alter the relative stabilities of transition states for prolyl cis-trans isomerization. Solvation free energies of local minima and transition states in chloroform and water were calculated using the conductor like polarizable continuum model at the HF/6-31+G(d) level of theory. The population of trans up-puckered conformations increases in the order Ac-Pro-NHMe < Ac-Hyp-NHMe < Ac-Flp-NHMe in chloroform and water. The increase in population for trans up-puckered conformations in solution is attributed to the increase in population for the polyproline-II-like conformations with up puckering. The barriers DeltaGct++ to prolyl cis-to-trans isomerization for Ac-Hyp-NHMe and Ac Flp-NHMe increase as the solvent polarity increases, as seen for Ac-Pro-NHMe. In particular, it was identified that the cis-trans isomerization proceeds through the clockwise rotation about the prolyl peptide bond for Ac-Hyp-NHMe and Ac-Flp NHMe in chloroform and water, as seen for Ac-Pro-NHMe. PMID- 16471765 TI - Proof of single-molecule sensitivity in surface enhanced Raman scattering (SERS) by means of a two-analyte technique. AB - A method is proposed to pin down unambiguous proof for single-molecule sensitivity in surface enhanced Raman spectroscopy (SERS). The simultaneous use of two analyte molecules enables a clear confirmation of the single (or few) molecule nature of the signals. This method eliminates most of the uncertainties associated with low dye concentrations in previous experiments. It further shows that single- or few-molecule signals are very common in SERS, both in liquids and on dry substrates. PMID- 16471764 TI - Peptide secondary structure folding reaction coordinate: correlation between uv raman amide III frequency, Psi Ramachandran angle, and hydrogen bonding. AB - We used UV resonance Raman (UVRR) spectroscopy to quantitatively correlate the peptide bond AmIII3 frequency to its Psi Ramachandran angle and to the number and types of amide hydrogen bonds at different temperatures. This information allows us to develop a family of relationships to directly estimate the Psi Ramachandran angle from measured UVRR AmIII3 frequencies for peptide bonds (PBs) with known hydrogen bonding (HB). These relationships ignore the more modest Phi Ramachandran angle dependence and allow determination of the Psi angle with a standard error of +/-8 degrees , if the HB state of a PB is known. This is normally the case if a known secondary structure motif is studied. Further, if the HB state of a PB in water is unknown, the extreme alterations in such a state could additionally bias the Psi angle by +/-6 degrees . The resulting ability to measure Psi spectroscopically will enable new incisive protein conformational studies, especially in the field of protein folding. This is because any attempt to understand reaction mechanisms requires elucidation of the relevant reaction coordinate(s). The Psi angle is precisely the reaction coordinate that determines secondary structure changes. As shown elsewhere (Mikhonin et al. J. Am. Chem. Soc. 2005, 127, 7712), this correlation can be used to determine portions of the energy landscape along the Psi reaction coordinate. PMID- 16471766 TI - Clouding: origin of phase separation in oppositely charged polyelectrolyte/surfactant mixed solutions. AB - The phase behavior in the system of cationic modified poly(vinyl alcohol) (CPVA) sodium dodecylsulfate (SDS)-water has been investigated. Samples were found phase separated near electroneutral mixing at CPVA concentrations < or =6%, while in a medium CPVA concentration of 7-12%, the phase separation disappeared and the system transformed into bluish homogeneous solution. At > or =13% CPVA concentrations, the mixed systems became colorless homogeneous. Preclouding phenomenon was observed in 5-8% CPVA-SDS mixed systems at an electroneutral mixing ratio. The addition of inorganic salts, such as Na2SO4, NaCl, NaBr, and NaSCN, could exclude the bluish and phase separation phenomenon that was found to be caused by the increase of clouding point in these systems. The clouding phenomenon was proven to be the origin of the phase separation in the CPVA-SDS mixed system. The ability for the inorganic salts to increase the clouding point follows the order of the Hofmeister series. PMID- 16471767 TI - Amyloid beta-peptide oligomerization in silico: dimer and trimer. AB - Soluble oligomers of Alzheimer's amyloid beta protein (Abeta) may act as effectors of neurotoxicity in early stages of Alzheimer's disease. Detailed information about the structure of Abeta in atomistic level and the dynamics of assembly of monomeric Abeta into oligomeric structures is rather elusive. We have performed replica exchange molecular dynamics (REMD) simulations on the formation of the dimer and trimer of Abeta10-35 peptide. We have observed spontaneous formation of several basic structural units that may act as a template or an intermediate for further aggregation of Alzheimer's Abeta protein. Various conformers, including interlocking structures of experimentally known bend double beta strands, are identified. PMID- 16471769 TI - Diffusion of long n-alkanes in silicalite. A comparison between neutron scattering experiments and hierarchical simulation results. AB - New quasi-elastic neutron scattering (QENS) data are presented for the self diffusion of normal alkanes up to hexadecane in the zeolite silicalite at low occupancy. Measured diffusivities at 300 K are higher than in Na-ZSM-5 and agree with the predictions of molecular dynamics and a hierarchical transition-state theory/Brownian dynamics conducted on the same systems. Activation energies for diffusion are around 5 kJ/mol for C1-C6 but rise with carbon number for C8-C16 to approximately 15 kJ/mol for C16. An explanation is suggested by the hierarchical simulation, which finds that C1-C6 tend to reside entirely within the interiors of single channel segments and diffuse through jumps across energetically less favorable channel intersections, while longer molecules straddle channel intersections and must undergo conformational rearrangements in order to diffuse. The agreement between QENS and simulations reflects that the two methods probe motion over the same small (nm) length scales. Self-diffusivities measured by pulsed field gradient nuclear magnetic resonance (PFG-NMR) in the same systems are appreciably lower. This is because PFG-NMR probes motion over significantly longer (microm) length scales and is therefore more sensitive to defects in the silicalite crystals. PMID- 16471768 TI - Comparative examination of the stability of semiconductor quantum dots in various biochemical buffers. AB - Due to their greater photostability compared to established organic fluorescence markers, semiconductor quantum dots provide an attractive alternative for the biolabeling of living cells. On the basis of a comparative investigation using differently sized thiol-stabilized CdTe nanocrystals in a variety of commonly used biological buffers, a method is developed to quantify the stability of such a multicomponent system. Above a certain critical size, the intensity of the photoluminescence of the nanocrystals is found to diminish with pseudo-zero-order kinetics, whereas for specific combinations of particle size, ligand, and buffer there appears to be no decay below this critical particle size, pointing out the necessity for thorough investigations of this kind in the view of prospect applications of semiconductor nanocrystals in the area of biolabeling. PMID- 16471771 TI - Transport diffusion of gases is rapid in flexible carbon nanotubes. AB - Molecular dynamics simulations of rigid, defect-free single-walled carbon nanotubes have previously suggested that the transport diffusivity of gases adsorbed in these materials can be orders of magnitude higher than any other nanoporous material (A. I. Skoulidas et al., Phys. Rev. Lett. 2002, 89, 185901). These simulations must overestimate the molecular diffusion coefficients because they neglect energy exchange between the diffusing molecules and the nanotube. Recently, Jakobtorweihen et al. have reported careful simulations of molecular self-diffusion that allow nanotube flexibility (Phys. Rev. Lett. 2005, 95, 044501). We have used the efficient thermostat developed by Jakobtorweihen et al. to examine the influence of nanotube flexibility on the transport diffusion of CH4 in (20,0) and (15,0) nanotubes. The inclusion of nanotube flexibility reduces the transport diffusion relative to the rigid nanotube by roughly an order of magnitude close to zero pressure, but at pressures above about 1 bar the transport diffusivities for flexible and rigid nanotubes are very similar, differing by less than a factor or two on average. Hence, the transport diffusivities are still extremely large compared to other known materials when flexibility is taken into account. PMID- 16471770 TI - Photoswitching behavior of a novel single molecular tip for noncontact atomic force microscopy designed for chemical identification. AB - A tripod molecule with an azobenzene arm was designed as a single molecular tip for noncontact atomic force microscopy (NC-AFM). The azobenzene moiety showed photoisomerization that enabled measurements of the same position of the sample by different tip apexes with different interactions. Photoswitching behavior of the molecule synthesized and adsorbed on Au surfaces was examined and reversible switching between the trans- and cis forms was successfully confirmed by NC-AFM measurements. PMID- 16471772 TI - Better chemistry through ceramics: the physical bases of the outstanding chemistry of ORMOSIL. AB - Twenty years after their invention, sol-gel organically modified silicates (ORMOSIL) are finding a number of impressive applications that range from efficient deliver of genes into mouse brains to self-ordered helices of interest to fields as diverse as optics, catalysis, molecular recognition, and chromatography. The physical bases of this mulifaceted chemistry, therefore, are of immense importance to scientists working toward new applications such as photovoltaics and catalysis that are crucially important in making sustainable global development. The purpose of this article is to provide a general picture of ORMOSIL's physical chemistry that will be useful in the creative development of new materials capable to solve a number of relevant open problems. PMID- 16471773 TI - Far-infrared characteristics of ZnS nanoparticles measured by terahertz time domain spectroscopy. AB - The optical and dielectric properties of ZnS nanoparticles are studied by use of terahertz time-domain spectroscopy (THz-TDS) over the frequency range from 0.3 to 3.0 THz. The effective medium approach combined with the pseudo-harmonic model of the dielectric response, where nanoparticles are embedded in the host medium, provides a good fit on the experimental results. The extrapolation of the measured data indicates that the absorption is dominated by the transverse optical mode localized at 11.6+/-0.2 THz. Meanwhile, the low-frequency phonon resonance of ZnS nanoparticles is compared with the single-crystal ZnS. The THz TDS clearly reveals the remarkable distinction in the low-frequency phonon resonances between ZnS nanoparticles and single-crystal ZnS. The results demonstrate that the acoustic phonons become confined in small-size nanoparticles. PMID- 16471774 TI - Surface chemistry of luminescent colloidal silicon nanoparticles. AB - Luminescent colloidal silicon particles are obtained by burning silane and slowly etching the product with hydrogen fluoride. Depending on their size, the particles emit red-orange or blue-green light with decay times between 28 micros and <0.1 micros, respectively. The quantum yield of the red luminescence is found to be 35%, i.e., much higher than the 7% previously reported, and the yield of the blue-green luminescence is 18%. The luminescence of the colloidal particles is quenched upon the attack of their surface by free radicals, oxygen-centered radicals being more efficient than carbon-centered ones. It is concluded from the dependence of the luminescence wavelength on the etching time and the dependence of the luminescence lifetime on the wavelength that the mechanism of the photoluminescence undergoes a change with decreasing particle size. The red luminescent particles exhibit amphiphilic properties, such as unusual wetting phenomena. This effect is understood in terms of the existence of few polar groups on the otherwise nonpolar surface of the particles, possibly Si-OH groups, which also act as centers of the red luminescence. PMID- 16471775 TI - Chemical reaction of nitric oxides with the 5-1DB defect of the single-walled carbon nanotube. AB - In this work, we applied a two-layered ONIOM (B3LYP/6-31G(d):UFF) method to study the reaction of nitric oxides with a 5-1DB defect on the sidewall of the single walled carbon nanotube (SWCNT). We have chosen a suitable ONIOM model for the calculation of the SWCNT based on the analyses of the frontier molecular orbitals, local density of states, and natural bond orbitals. Our calculations clearly indicate that the 5-1DB defect is the chemically active center of the SWCNT. In the reaction of nitric oxides with the defected SWCNT, the 5-1DB defect site can capture a nitrogen atom from nitric oxides, yielding the N substitutionally doped SWCNT. We have explored the reaction pathway in detail. Our work verifies the chemical reactivity of the 5-1DB defects of the SWCNTs, indicates that the 5-1DB defect is a possible site for the functionalization of the SWCNTs, and demonstrates a possible way to fabricate position controllable substitutionally doped SWCNTs with a low doping concentration under mild conditions via some simple chemical reactions. PMID- 16471776 TI - Morphology-controllable synthesis and characterization of single-crystal molybdenum trioxide. AB - Molybdenum trioxide nanobelts and prism-like particles with good crystallinity and high surface areas have been prepared by a facile hydrothermal method, and the morphology could be controlled by using different inorganic salts, such as KNO3, Ca(NO3)2 , La(NO3)3, etc. The possible growth mechanism of molybdenum trioxide prism-like particles is discussed on the basis of the presence of H+ and the modification of metal cations. The as-prepared nanomaterials are characterized by means of powder X-ray diffraction (PXRD), field-emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), high-resolution TEM (HRTEM), Fourier transformation infrared spectroscopy (FT IR), X-ray photoelectron spectroscopy (XPS), and ultraviolet and visible spectroscopy (UV-vis). TEM and HRTEM micrographs show that the molybdenum trioxide nanobelts and prism-like particles have a relatively high degree of crystallinity and uniformity. BET specific surface areas of the as-prepared molybdenum trioxide nanocrystals are 67-79 m2 g-1. XPS analysis indicates that the hexavalent molybdenum is predominant in the nanocrystals. UV-vis spectra reveal that the direct band gap energy of the annealed molybdenum trioxide prism like particles shows a pronounced blue shift compared to that of bulk MoO3 powder. Interestingly, molybdenum trioxide nanobelts exhibit a red shift under this condition. PMID- 16471777 TI - A small paramagnetic platinum cluster in an NaY zeolite: characterization and hydrogen adsorption and desorption. AB - A well-defined cluster containing 12 equivalent platinum atoms was prepared by ion exchange of an NaY zeolite, followed by hydrogen reduction. It was characterized by electron paramagnetic resonance (EPR) spectroscopy, hyperfine sublevel correlation (HYSCORE), and theoretical calculations. Combing the results of the experiments with density functional calculations, the likely structure of this cluster is icosahedral Pt13Hm, possibly with a low positive charge. The adsorbed H/D on the Pt cluster surface can be exchanged reversibly at room temperature. From H/D desorption experiments, an H2 binding energy of 1.36 eV is derived, in reasonable agreement with the calculated value but clearly larger than that for a (111) Pt single-crystal surface, revealing a finite size effect. While the hydrogen-covered cluster should clearly be regarded as a molecule, it is conceivable that the cluster adopts metallic character upon hydrogen desorption. It is likely that up to m=30 H atoms bind to this cluster with 12 surface atoms, which has important implications for the determination of the dispersion of small Pt catalyst particles by hydrogen chemisorption. Calculations as well as experiments give evidence of an interesting magnetic behavior with high-spin states playing a prominent role. There are strong indications that a reservoir of EPR silent but structurally similar clusters exists which can partly be converted to EPR visible species by H/D exchange or by gas adsorption. PMID- 16471778 TI - Aggregates in solution of binary mixtures of amphiphilic diblock copolymers with different chain length. AB - The polydispersity effect of amphiphilic AB diblock copolymers on the self assembled morphologies in solution has been investigated by the real-space implementation of self-consistent field theory (SCFT) in two dimensions (2D). The polydispersity is artificially obtained by mixing binary diblock copolymers where the hydrophilic or hydrophobic blocks are composed of two different lengths while the other block length is kept the same. The main advantage is that this simple polydispersity can easily distinguish the difference of aggregates in the density distribution of long and short block length intuitionally and quantitatively. The morphology transition from vesicles to micelles is observed with increasing polydispersity of copolymers due to the length segregation of copolymers. For polydisperse hydrophilic or hydrophobic blocks, the short blocks tend to distribute at the interfaces between hydrophilic and hydrophobic blocks while the long blocks stretch to the outer space. More specifically, by quantitatively taking the sum of all the concentration distribution of long and short chains over the inside and outside surface areas of the vesicle, it is found that long blocks prefer to locate on the outside surface of the vesicle while short ones prefer the inside. Such length segregation leads to large curvature of the aggregate, thus resulting in the decrease of the aggregate size. PMID- 16471779 TI - High-density assembly of gold nanoparticles on multiwalled carbon nanotubes using 1-pyrenemethylamine as interlinker. AB - In this article, we describe the formation of carbon nanotube (CNT)-gold nanoparticle composites in aqueous solution using 1-pyrenemethylamine (Py-CH2NH2) as the interlinker. The alkylamine substituent of 1-pyrenemethylamine binds to a gold nanoparticle, while the pyrene chromophore is noncovalently attached to the sidewall of a carbon nanotube via pi-pi stacking interaction. Using this strategy, gold nanoparticles with diameters of 2-4 nm can be densely assembled on the sidewalls of multiwalled carbon nanotubes. The formation of functionalized gold nanoparticles and CNT-Au nanoparticle composites was followed by UV-vis absorption and luminescence spectroscopy. After functionalization of gold nanoparticles with 1-pyrenemethylamine, the distinct absorption vibronic structure of the pyrene chromophore was greatly perturbed and its absorbance value was decreased. There was also a corresponding red shift of the surface plasmon resonance (SPR) absorption band of the gold nanoparticles after surface modification from 508 to 556 nm due to interparticle plasmon coupling. Further reduction of the pyrene chromophore absorbance was observed upon formation of the CNT-Au nanoparticle composites. The photoluminescence of 1-pyrenemethylamine was largely quenched after attaching to gold nanoparticles; formation of the CNT-Au nanoparticle composites further lowered its emission intensity. The pyrene fluoroprobe also sensed a relatively nonpolar environment after its attachment to the nanotube surface. The present approach to forming high-density deposition of gold nanoparticles on the surface of multiwalled carbon nanotubes can be extended to other molecules with similar structures such as N-(1-naphthyl)ethylenediamine and phenethylamine, demonstrating the generality of this strategy for making CNT Au nanostructure composites. PMID- 16471780 TI - Generation of hydrophilic, bamboo-shaped multiwalled carbon nanotubes by solid state pyrolysis and its electrochemical studies. AB - A simple, efficient, and novel method was developed for the direct preparation of hydrophilic, bamboo-shaped carbon nanotubes by the pyrolysis of ruthenium(III) acetylacetonate in a Swagelock cell is reported. The obtained product exhibits mostly bamboo-shaped, straight, periodic twisted, multiwalled carbon nanotubes possessing diameters of 50-80 nm and lengths of around 10 microm. The pyrolyzed product was characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), high resolution TEM (HRTEM), Fourier transform infrared (FT-IR), X-ray photoelectron spectroscopy (XPS), micro-Raman, and cyclic voltammetric techniques. HRTEM studies showed that the walls of bamboo-shaped carbon nanotubes consisted of oblique grapheme planes with respect to the tube axis. The interlayer spacing between two graphitic layers was found to be 0.342 nm. XPS measurements have suggested that as-prepared carbon nanotubes consist the surface functional groups on the surface of carbon nanotubes. The electrochemical properties of synthesized carbon nanotubes have been evaluated. Thermogravimetric analysis (TGA), IR, and cyclic voltammetric studies showed the presence of oxygen functionalities. Raman studies revealed the presence of disorder in the graphitic carbon and the presence of exposed edge plane defects in the generated carbon nanotubes for influencing the surface behavior and electrochemical properties. The electrochemical behavior of electrodes made of bamboo-shaped carbon nanotubes served for an oxygen reduction reaction. PMID- 16471781 TI - Crystal structure of zeolite MCM-68: a new three-dimensional framework with large pores. AB - The crystal structure of the aluminosilicate MCM-68 was solved from synchrotron powder diffraction data by the program FOCUS. The unit cell framework contains Si100.6Al11.4O224. This material crystallizes in space group P42/mnm, where, after Rietveld refinement, a=18.286(1) A and c=20.208(2) A. A three-dimensional framework is found that contains continuous 12-ring channels and two orthogonal, intersecting, undulating 10-ring channels. Rietveld refinement of the model coordinates optimizes the framework geometry, to match the observed intensity profile by Rwp=0.1371, R(F2)=0.1411. It is not possible to determine the location of approximately 0.84 K+ cations remaining in the unit cell after the material is steamed and then dehydrated. The framework model also successfully predicts observed electron diffraction data in two projections, and the tetragonal projection can be determined independently from these data by direct methods. The calculated density of the framework structure is 1.66 g/cm3, and the T-site framework density is 16.6 T/1000 A3. PMID- 16471782 TI - Surface plasmon resonance of silver nanoparticles on vanadium dioxide. AB - The localized surface plasmon resonance (SPR) spectrum of silver nanoparticles fabricated on a thermochromatic film, vanadium dioxide (VO2), is studied in this paper. Owing to the temperature-dependent dielectric function of VO2, the SPR band dramatically exhibits temperature dependence in the range of 30-80 degrees C. The peak extinction wavelength, lambda(SPR), blueshifts as temperature increases and reversibly redshifts as temperature decreases. The shift magnitude (DeltalambdaSPR) is strongly dependent on the silver mass thickness, dm; a value of 50 nm of DeltalambdaSPR is achieved for particles (mean diameter 51 nm) with dm=2 nm while a value of 250 nm is achieved for particles (mean diameter 133 nm) with dm=10 nm. Beyond the SPR band, it is interesting to find that the spectral line shape of silver particles is dominated by the imaginary part of the dielectric function of VO2. These results can be interpreted based on dynamical Maxwell-Garnett theory. PMID- 16471783 TI - Determination of ionic conductivity and its impact on proton diffusion model for nickel hydroxide. AB - The ionic conductivity is an important but previously ignored aspect for the nickel hydroxide used in alkaline batteries. With a specially designed device, the ionic conductivity is determined for single beads of spherical nickel hydroxide in KOH solutions. The apparent ionic conductivity is found on the order of 10(-3)-10(-2) S cm-1 in 6 M KOH and to change with the conductivity of the solution in which the bead is immersed. The ionic conductivity of the bead can be mainly attributed to the electrolyte absorbed in the bead. On the basis of these findings, the dual structure model for proton diffusion in spherical nickel hydroxide is refined by specifying nanoparticles to be the component showing a large apparent proton diffusion coefficient (on the order of 10(-7) cm2 s-1). This refined model is able to interpret the main features of the diffusion coefficients reported in the literature, including the unusually large scattering (up to 6 orders of magnitude) and inconsistency in the dependence of proton diffusion coefficient on the state of the charge. Besides, this refined model is supported by the influence of bulk KOH concentration on chronoamperometry and transmission electron microscopy observations. PMID- 16471784 TI - Optical analysis of the light emission from porous silicon: a hybrid polyatom surface-coupled fluorophor. AB - The most extensive data set yet generated correlating photoluminescence excitation (PLE) and photoluminescence (PL) spectra is presented for aged (equilibrated) porous silicon (PS) samples. The observed features, which are temperature independent over the range 10-300 K, show a detailed correlation with the results of photoacoustic spectroscopy (PAS) and with molecular electronic structure calculations. The observed energy level patterns are reproduced in the photoabsorption (PA) of PS films released after the etching of a silicon wafer. It is concluded that the energy level pattern found for the photoluminescing surface of PS results from a structure which is neither uniquely molecule- or bulk-like but represents a hybrid form for which the density of states associated with a polyatomic vibrationally excited surface-bound fluorophor dominates the nature of the observed features which are not those of a semiconductor. These fluorophor features are broadened and shifted to lower excitation energy as a result of the intimate presence of the silicon surface to which the fluorophor is bound. The dominance of the surface-bound fluorophor accounts for the temperature independent PLE and PL features. The observed spectral features are thus suggested to be the result of a strong synergistic interaction in which the silicon surface influences the location of surface-bound fluorophor excited states whereas the nature of the vibrationally excited surface-bound fluorophor coupling to the silicon surface provides the mechanism for an enhanced vibronic structure dominated interaction and energy transfer. The observed PLE, PL, PAS, and PA measurements are found to be consistent with previous photovoltaic and photoconductivity measurements, correlating well with a surface-bound oxyhydride like emitter. This study suggests the important role that the overtone structure of a molecule bound to a surface can play as one forms a hybrid system. PMID- 16471785 TI - Recombination dynamics of CdTe/CdS core-shell nanocrystals. AB - The recombination dynamics of zinc-blende-type, deep-red emitting CdTe/CdS core shell nanocrystals is studied over a wide temperature range. Two characteristic decay regimes are found: a temperature-dependent decay component of a few nanoseconds and a long-living temperature-independent component of approximately 315 ns. The average decay time of the exciton states changes from 20 to 5ns when the temperature is increased from 15 to 295 K. At low temperatures, the observed decay behavior is assigned to thermally induced population and decay of the allowed exchange-split exciton states. At temperatures above T>100 K, nonradiative decay channels involving phonons start to contribute to the exciton recombination. The observed broad distribution in decay times, monitored by stretched exponential fitting functions, we explain by variations in the electron hole overlap caused by a partly incomplete CdTe/CdS core-shell structure and the nearly energy-degenerated bright and dark state superposition. PMID- 16471786 TI - A novel carbon nanotube structure formed in ultra-long nanochannels of anodic aluminum oxide templates. AB - We report the fabrication of a novel carbon structure consisting of uniform carbon nanotubes formed in the nanochannels of anodic aluminum oxide (AAO) templates, with the surface side open and connected by a uniform carbon sheet. The uniformity of the fabricated CNT arrays, plus the carbon film on the AAO surface interconnecting the open ends of all CNTs, constitute the major characteristics unique to our carbon structures. Some potential applications of such structures are noted. PMID- 16471787 TI - Synthesis and optical property of beta-brass colloid. AB - Cu/Zn beta-phase alloy (beta-brass) colloid was successfully synthesized by reducing the organometallic complexes in the organic liquid phase. It was confirmed by X-ray diffraction that the material formed pure beta-phase. The morphology for the beta-brass particles was observed using transmission electron microscopy (TEM). The dispersion of the particle diameter was almost single modal, and the diameter was estimated at 8+/-2 nm. A clear peak around 520 nm was seen in the optical absorption spectrum for the beta-brass colloid, and it is attributed to the surface plasmon absorption. We also simulated the optical absorption spectrum using the bulk dielectric constant for beta-brass. The simulation qualitatively reproduced the experimental data well. PMID- 16471788 TI - Dye-sensitized solar cells based on a single-crystalline TiO2 nanorod film. AB - Highly crystalline TiO2 nanorods with lengths of 100-300 nm and diameters of 20 30 nm have been synthesized by a hydrothermal process in a cetyltrimethylammonium bromide surfactant solution. The microstructure measured by X-ray diffraction and high-resolution transmission electron microscopy was a pure highly crystalline anatase phase with a long nanorod shape. The addition of a triblock copolymer poly(ethylene oxide)100-poly(propylene oxide) 65-poly(ethylene oxide)100 (F127) decreased the length of the nanorods and kept the rod shape of the particles even after sintering at high temperatures. The rod shape kept under high calcination temperatures contributed to the achievement of the high conversion efficiency of light-to-electricity as discussed in the paper. A high conversion efficiency of light-to-electricity of 7.29% was obtained with the TiO2 single-crystalline anatase nanorod cell. PMID- 16471789 TI - Light-emission and excited-state dynamics in Tm2+ doped CsCaCl3, CsCaBr3, and CsCaI3. AB - The light-emission and photophysical properties of CsCaCl3:Tm2+ (1.04%), CsCaBr3:Tm2+ (0.48%), and CsCaI3:Tm2+ (0.76%) are presented. We find that Tm2+ is a multiple emitter under 21,834 cm-1 laser excitation at low temperatures in all three compounds. Several distinct types of emission are observed and characterized: sharp and long-lived 4f-4f emission in the infrared (IR) and up to four broad and fast decaying emission bands in the near-IR and visible, originating from the 4f-5d states of Tm2+. The optical spectroscopic properties of the samples are compared, and we find that the measured differences in the relative intensities and the shifts in the position of the emissions can be related to the chemical influence on the absorption and emission properties of Tm2+. Thus, it nicely illustrates the principle of chemical variation on the optical spectroscopic properties. An investigation of the temperature dependence of the luminescence yields important information about the dynamics of the excited states. The interplay and competition between radiative and nonradiative pathways is explained and modeled using a single configurational coordinate approach. PMID- 16471790 TI - Experimental evidence for the triplet-like spin state appearing in ground-state singlet biradicals as a key feature for generalized ferrimagnetic spin alignment. AB - The authors have previously proposed a theoretical model for exotic spin alignment in organic molecular assemblages: The alternating chain of organic biradicals in a singlet (Sb=0) ground state and monoradicals with S=1/2 has a ferrimagnetic ground state for the whole chain, which has been termed generalized ferrimagnetism. An important feature of the generalized ferrimagnetic spin alignment has been found in the deviation of the expectation value Sb2 of the biradical spin from zero. Even a triplet-like spin state Sb2=2 (Sb=1) has been predicted in the theoretical calculations. In this study, we have found experimental evidence for the pseudo-triplet state appearing in the ground-state singlet biradical of a real open-shell compound. At first, we have demonstrated from theoretical calculations that the singlet biradical has Sb2=2 (Sb=1) in a molecular pair with an S=1 metal ion as well as with the S=1/2 monoradical. The pseudo-triplet state of the biradical affords a singlet state of the whole system of the biradical-metal ion pair, which is readily detectable in experiments for verifying the theoretical prediction. As a model compound for the biradical-metal ion pair, a transition metal complex, [(bnn)(Ni(hfac)2)1.5(H2O)] (1), has been synthesized from a nitronyl nitroxide-based ground-state singlet biradical bnn and Ni(hfac)2. From X-ray crystallographic analyses, the compound contains a molecular pair of bnn and Ni(hfac)2, which serves as a model system under the above theoretical studies. It has been found from the analysis of the temperature dependence of magnetic susceptibility that the bnn-Ni(hfac)2 pair has the singlet (S=0) ground state. The singlet ground state of the pair results from an antiparallel coupling of the pseudo-triplet of the biradical and the S=1 spin on the Ni ion. The pseudo-triplet state in the ground-state singlet biradical has thus been verified experimentally, which is crucially important to realize the generalized ferrimagnetic spin alignment. PMID- 16471791 TI - Tailoring (n,m) structure of single-walled carbon nanotubes by modifying reaction conditions and the nature of the support of CoMo catalysts. AB - The (n,m) population distribution of single-walled carbon nanotubes obtained on supported CoMo catalysts has been determined by photoluminescence and optical absorption. It has been found that the (n,m) distribution can be controlled by varying the gaseous feed composition, the reaction temperature, and the type of catalyst support used. When using CO as a feed over CoMo/SiO2 catalysts, increasing the synthesis temperature results in an increase in nanotube diameter, without a change in the chiral angle. By contrast, by changing the support from SiO2 to MgO, nanotubes with similar diameter but different chiral angles are obtained. Finally, keeping the same reaction conditions but varying the composition of the gaseous feed results in different (n,m) distribution. The clearly different distributions obtained when varying catalysts support and/or reaction conditions demonstrate that the (n,m) distribution is a result of differences in the growth kinetics, which in turn depends on the nanotube cap metal cluster interaction. PMID- 16471792 TI - Supra-aggregates of fiber-forming anisotropic molecules. AB - In this paper, the self-organization of fiber-forming anisotropic molecules is inspected both theoretically and experimentally. In the first part, a theoretical model which extends the de Gennes theory of thin films to assemblies of strongly anisotropic molecules is reported. The model predicts that solid supported thin films made up of fiber-forming discotic molecules can grow with both tangential and radial arrangement of the fibers, respectively leading to the formation of compact and holed supra-aggregates. These last systems form according to the following picture. The tangential growth minimizes the number of unfavorable free ends but introduces elastic strain especially in the central region of the aggregate. To reduce the elastic strain, some molecules are displaced from the central region toward the periphery of the growing aggregate, producing a localized well. In the second part of the paper, we experimentally face the above issue by depositing a strongly anisotropic disk-shaped molecule (rhodamine 123) onto different solid substrates through a spin coating procedure. By employing scanning force microscopy (SFM), the formation of thermodynamically favored fiberlike supramolecules as well as of compact and holed submicron-sized supra aggregates has been demonstrated. The observed phenomena have been found to depend on the interplay of different parameters such as molecular concentration, evaporation time, and substrate composition. As main features, both theory and experiments show that holed supra-aggregates are more stable beyond a critical aggregation size and that the formation of holes is favored at high supersaturation. The theory seems valuable in extending previous dewetting models developed for fluid films with isotropic interaction forces. PMID- 16471793 TI - Optical absorption and valence band photoemission from uncapped CdTe nanocrystals. AB - CdTe nanocrystals have been successfully fabricated by a mechanical alloying process. X-ray diffraction (XRD) patterns demonstrate that a single-phase CdTe compound with a zinc blende structure has been formed after ball milling elemental Cd and Te mixture powders for 27 h. The large broadening effect for the width of the {111} diffraction peak of uncapped CdTe nanocrystals on smaller size was observed in slowly scanned XRD patterns. The X-ray photoelectron spectrum was used to study the surface of the uncapped CdTe nanocrystals within both core level and valence band regions. The presence of tellurium oxide film on the surface of the uncapped CdTe nanocrystals has been detected in the X-ray photoelectron spectrum of the Te 3d core level, which was comparable to the observed amorphous oxide thin layer on the surface of uncapped CdTe nanocrystals in a high resolution transmission electron microscopy (HRTEM) image. The energy of the valence band maximum for uncapped CdTe powders blue shifts to the higher energy side with smaller particle sizes. In UV-visible optical absorption spectra of the suspension solution containing uncapped CdTe nanocrystals, the absorption peaks were locating within the ultraviolet region, which shifted toward the higher energy side with prolonged ball milling time. Both blue shifts of valence band maximum energy and absorption peaks with decreasing particle size provide a unique pathway to reveal the quantum confinement effect of uncapped CdTe nanocrystals. PMID- 16471794 TI - Solid-state NMR spectroscopy of anionic framework aluminophosphates: a new method to determine the al/p ratio. AB - A series of anionic framework aluminophosphates, with different Al/P ratios, have been investigated by various solid-state NMR techniques, including 27Al, 31P magic angle spinning (MAS), 27Al-->31P cross polarization (CP), 27Al{31P} rotational echo double resonance (REDOR), and 31P{27Al} transfer of population double resonance (TRAPDOR). Different Al coordinations (AlO4b, AlO5b, and AlO6b) and P coordinations (PO4b, PO3bOt, PO2bO2t, and PObO3t), where b represents bridging oxygens and t represents terminal oxygens, can be unambiguously determined based on the solid-state NMR spectroscopy. Furthermore, a new method to determine the Al/P ratio of open-framework aluminophosphates has been established, which is useful for the understanding of unknown aluminophosphate structures. PMID- 16471795 TI - Anomalously large formula unit volume and its effect on the thermal behavior of LiBF4. AB - The crystal structure of LiBF4 has been determined by single-crystal X-ray diffraction measurements. LiBF4 crystallizes as a merohedral twin in the trigonal space group P3(1)21 with a=4.892(5) A, c=11.002(12) A, V=228.0(4) A3, and Z=3 at 200 K. The twin is generated by a 2-fold rotation about the [10] direction. The lithium cation is coordinated by four fluorine atoms in a distorted tetrahedral manner, wherein two Li-F distances of 1.862(5) and 1.846(5) A are observed. The formula unit volume (FUV=V/Z) of 77.9 A3 for LiBF4 at 298 K is considerably larger than 72.7 A3 for NaBF4 and 72.5 A3 for AgBF4, despite the smaller size of Li+, indicating loose ionic packing of LiBF4. The thermodynamic evaluation of the decomposition temperature for LiBF4 was performed using the empirical relationship between the standard entropy and the FUV obtained. The results indicate that the large FUV of LiBF4 contributes to its higher decomposition temperature compared to that of LiPF6. PMID- 16471796 TI - Anion complexation by calix[3]thieno[1]pyrrole: the medium effect. AB - The interaction of calix[3]thieno[1]pyrrole, 1, and halide and dihydrogen phosphate anions in a variety of solvents (acetonitrile, propylene carbonate, N,N dimethylformamide, and dimethyl sulfoxide) has been investigated through 1H NMR, conductance measurements, and titration calorimetry. 1H NMR measurements reveal the sites of interaction of the ligand with the anions in CD3CN while the composition of the complex was determined through conductance measurements. A quantitative assessment of anion-ligand interactions is provided. Thus the thermodynamics of complexation of 1 with halide and dihydrogen phosphate anions in dipolar aprotic media at 298.15 K is reported. These data are interpreted in terms of the thermodynamics of transfer of reactants and product from a reference solvent (acetonitrile) to other solvents. The crucial role played by the solvent on the ability of the ligand to interact with anions and on the composition of the complex is demonstrated. PMID- 16471798 TI - Low-temperature metallic alloying of copper and silver nanoparticles with gold nanoparticles through digestive ripening. AB - We describe a remarkable and simple alloying procedure in which noble metal intermetallic nanoparticles are produced in gram quantities via digestive ripening. This process involves mixing of separately prepared colloids of pure Au and pure Ag or Cu particles and then heating in the presence of an alkanethiol under reflux. The result after 1 h is alloy nanoparticles. Particles synthesized according to this procedure were characterized by UV-vis spectroscopy, EDX analysis, and high-resolution electron microscopy, the results of which confirm the formation of alloy particles. The particles of 5.6+/-0.5 nm diameter for Au/Ag and 4.8+/-1.0 nm diameter for Cu/Au undergo facile self-assembly to form 3 D superlattice ordering. It appears that during this digestive ripening process, the organic ligands display an extraordinary chemistry in which atom transfer between atomically pure copper, silver, and gold metal nanoparticles yields monodisperse alloy nanoparticles. PMID- 16471799 TI - Linear nanostructure formation of aldehydes by self-directed growth on hydrogen terminated silicon(100). AB - The self-directed growth of organic molecules on silicon surfaces allows for the rapid, parallel production of hybrid organic-silicon nanostructures. In this work, the formation of benzaldehyde- and acetaldehyde-derived nanostructures on hydrogen-terminated H-Si(100)-2x1 surface is studied by scanning tunneling microscopy in ultrahigh vacuum and by quantum mechanical methods. The reaction is a radical-mediated process that binds the aldehydes, through a strong Si-O covalent bond, to the surface. The aldehyde nanostructures are generally composed of double lines of molecules. Two mechanisms that lead to double line growth are elucidated. PMID- 16471797 TI - Multipole plasmon resonances in gold nanorods. AB - The optical properties of gold rods electrochemically deposited in anodic aluminum oxide templates have been investigated. Homogeneous suspensions of rods with an average diameter of 85 nm and varying lengths of 96, 186, 321, 465, 495, 578, 641, 735, and 1175 nm were fabricated. The purity and dimensions of these rod nanostructures allowed us to observe higher-order multipole resonances for the first time in a colloidal suspension. The experimental optical spectra agree with discrete dipole approximation calculations that have been modeled from the dimensions of the gold nanorods. PMID- 16471800 TI - Structure investigation of Ag(111)(radical7x radical7)R19 degrees -SCH3 by X-ray standing waves: a case of thiol-induced substrate reconstruction. AB - The structure of the Ag(111)(radical7x radical7)R19 degrees-CH3S surface phase, formed by interaction of Ag(111) with gas-phase dimethyl disulfide, (CH3S)2, has been investigated by normal-incidence X-ray standing wave (NIXSW) analysis, using (111), (11), and (200) Bragg reflections. The resulting NIXSW structural parameter values clearly exclude any simple overlayer adsorption model on an Ag unreconstructed surface. A reconstructed surface model is proposed that is consistent with the NIXSW measurements and with previous scanning tunneling microscopy results. This comprises a near-hexagonal Ag surface layer with an Ag density of only 3/7 that of the underlying substrate layers; the methanethiolate molecules are adsorbed into 3-fold coordinated hollow sites on this open layer. The results are discussed in the context of the very limited published studies of longer alkyl chain thiolates on Ag(111). PMID- 16471802 TI - Molecular aggregation of rhodamine dyes in dispersions of layered silicates: influence of dye molecular structure and silicate properties. AB - The molecular aggregation of six rhodamine dyes (rhodamine 560, B, 3B, 19, 6G, 123) in layered silicate (saponite and fluorohectorite) dispersions was investigated by using visible (vis) spectroscopy. The dye molecular aggregation was influenced by the properties of both the silicates and the dyes themselves. The layer charge of the silicates enhanced the molecular aggregation of the hydrophilic, cationic dyes. The presence of a carboxyl acid group in the dye molecules inhibited adsorption of the dyes on the surface of fluorohectorite, a silicate with a high charge density. A lower or no adsorption could be observed by vis spectroscopy. Strong association of the dyes to the silicate surface led to remarkable changes in the dye spectra, mainly due to the molecular aggregation. Dye assemblies initially formed after mixing the dye solutions with silicate dispersions were unstable. Decomposition of the dye molecular assemblies, and the formation of new species or molecular aggregate rearrangements, were studied on the bases of time-difference spectra. The reaction pathways were specific, not only for the dyes, depending upon their molecular structure and properties, but also on the silicate substrates. PMID- 16471801 TI - Fabrication of electroactive layer-by-layer films of myoglobin with gold nanoparticles of different sizes. AB - Alternate adsorption of oppositely charged myoglobin (Mb) and gold nanoparticles with different sizes were used to assemble {Au/Mb}n layer-by-layer films on solid surfaces by electrostatic interaction between them. The direct electrochemistry of Mb was realized in {Au/Mb}n films at pyrolytic graphite (PG) electrodes, showing a pair of well-defined, nearly reversible cyclic voltammetry (CV) peaks for the Mb heme FeIII/FeII redox couple. Quartz crystal microbalance (QCM), electrochemical impedance spectroscopy (EIS), and CV were used to monitor or confirm the growth of the films. Compared with other Mb layer-by-layer films with nonconductive nanoparticles or polyions, {Au/Mb}n films showed much improved properties, such as smaller electron-transfer resistance (Rct) measured by EIS with Fe(CN)3-/4- redox probe, higher maximum surface concentration of electroactive Mb (Gamma*max), and better electrocatalytic activity toward reduction of O2 and H2O2, mainly because of the good conductivity of Au nanoparticles. Because of the high biocompatibility of Au nanoparticles, adsorbed Mb in the films retained its near native structure and biocatalytic activity. The size effect of Au nanoparticles on the electrochemical and electrocatalytic activity of Mb in {Au/Mb}n films was investigated, demonstrating that the {Au/Mb}n films assembled with smaller-sized Au nanoparticles have smaller Rct, higher Gamma*max, and better biocatalytic reactivity than those with larger size. PMID- 16471803 TI - An investigation of gold adsorption from a binary mixture with selective mesoporous silica adsorbents. AB - Gold-selective adsorbents were prepared from mesoporous MCM-41 silica by grafting organic amine groups (i.e., RNH2, R2NH, and R3N; R=propyl). NH2-MCM-41, NRH-MCM 41, and NR2-MCM-41 displayed strong affinity for gold and at 1 mmol/g loading adsorbed 0.40, 0.33, and 0.20 mmol/g of gold. Copper and nickel were not adsorbed on these adsorbents. Grafting surface chemical moieties introduces heterogeneity on an otherwise uniform MCM-41 pore surface and metal adsorption is best described by the Freundlich adsorption model. A series of binary adsorption equilibrium studies with NH2-MCM-41 containing 2.2 mmol RNH2/g shows that NH2-MCM 41 adsorbs only gold from solutions containing copper and nickel with an adsorption capacity of 0.6 mol of Au/mol of RNH2 (1.1 mmol of Au/g of NH2-MCM 41). Copper and nickel were not adsorbed by NH2-MCM-41 regardless of the solution concentration, composition, and pH (i.e., 2 to 4) in the presence of gold. The LeVan and Vermeulen adsorption model based on a single component Freundlich isotherm and corrected for the anion effect accurately predicted the binary adsorptions. The adsorbed gold was completely recovered by a simple acid wash and the recovered gold solution is 99% pure. The regenerated NH2-MCM-41 remained 100% selective for gold removal and exhibited the same adsorption capacity even after several uses. PMID- 16471804 TI - Influence of isotherm inflection on the loading dependence of the diffusivities of n-hexane and n-heptane in MFI zeolite. Quasi-elastic neutron scattering experiments supplemented by molecular simulations. AB - Quasi-Elastic Neutron Scattering (QENS) experiments were carried out to determine (a) Fick diffusivity, D (b) self-diffusivity, Dself, and (c) 1/Gamma, the inverse of the thermodynamic correction factor, for n-hexane (nC6) and n-heptane (nC7) in MFI zeolite (all silica silicalite-1) at 300 K for a variety of loadings. These experimental results are compared with configurational-bias Monte Carlo (CBMC) and molecular dynamics (MD) simulations of, respectively, the adsorption isotherms and diffusivities. For n-hexane, the CBMC simulated isotherm shows a slight inflection at a loading=4 molecules per unit cell; this inflection manifests, also, in the loading dependence of 1/Gamma, obtained from QENS. The trend in the loading dependence of the Fick D and Dself of nC6 obtained from QENS matches the MD simulation results. For nC7 the CBMC simulated isotherm shows a strong inflection at a loading=4 molecules per unit cell. At this loading=4, 1/Gamma tends to zero and there is a very good match between QENS and molecular simulations for the loading dependence of 1/Gamma. Both MD simulations and QENS data on the Fick diffusivity shows a sharp maximum at a loading in the region of=4. For both nC6 and nC7 the simulated values of diffusivity are about an order of magnitude higher than those determined from QENS. PMID- 16471805 TI - Electrophoretic mobility, zeta potential, and fixed charge density of bovine knee chondrocytes, methyl methacrylate-sulfopropyl methacrylate, polybutylcyanoacrylate, and solid lipid nanoparticles. AB - The electrophoretic mobility and zeta potential of bovine knee chondrocytes (BKCs), methyl methacrylate-sulfopropyl methacrylate (MMA-SPM) nanoparticles (NPs), polybutylcyanoacrylate (PBCA) NPs, and solid lipid nanoparticles (SLNs) were investigated under the influences of Na+, K+, and Ca2+ with various ionic strengths. The fixed charge density in the surface layers of the four biocolloidal particles was estimated from the experimental mobility of capillary electrophoresis with a theory of soft charged colloids. The results revealed that, for a specific cationic species, the absolute values of the electrophoretic mobility, the zeta potential, and the fixed charge density decreased with an increase in ionic strength. For a constant ionic strength, the effect of ionic species on the reduction in the absolute values of the electrophoretic mobility, the zeta potential, and the fixed charge density followed the order Na+>K+>Ca2+ for the negatively charged BKCs, MMA-SPM NPs, and SLNs. The reverse order is true for the positively charged PBCA NPs. PMID- 16471806 TI - Friction of mixed and single-component aromatic monolayers in contacts of different adhesive strength. AB - Friction force microscopy has been used to study single-component and mixed self assembled monolayers of aminothiophenol and thiophenol on gold. The friction forces and transition pressures of mixed monolayers were intermediate to the ones of single-component monolayers, and varied systematically with composition. The strength of the adhesion was altered by working in dry N2 gas or in ethanol. In all systems studied, low adhesion (in ethanol) resulted in a linear dependence of the friction on load already at low loads, whereas high adhesion (in dry N2) gave an apparent area-dependence. However, for a given monolayer composition, similar transition pressures were observed in dry N2 and in ethanol, suggesting that the overall monolayer structure was not strongly altered by the presence of ethanol. Similar observations were made for very close-packed monolayers of octadecanethiol. PMID- 16471807 TI - Photocatalytic activity of R3MO7 and R2Ti2O7 (R=Y, Gd, La; M=Nb, Ta) for water splitting into H2 and O2. AB - The photocatalytic activities of R3MO7 and R2Ti2O7 (R=Y, Gd, La; M=Nb, Ta) strongly depended on the crystal structure. Overall, photocatalytic water splitting into H2 and O2 proceeded over La3TaO7 and La3NbO7, which have an orthorhombic weberite structure, Y2Ti2O7 and Gd2Ti2O7, which have a cubic pyrochlore structure, and La2Ti2O7, which has a monoclinic perovskite structure. All of these materials are composed of a network of corner-shared octahedral units of metal cations (TaO6, NbO6, or TiO6); materials without such a network were inactive. The octahedral network certainly increased the mobility of electrons and holes, thereby enhancing photocatalytic activity. PMID- 16471809 TI - Molecular dynamics simulations of the effect of the composition of calcium alumino-silicate intergranular films on alumina grain growth. AB - Molecular dynamics simulations were performed to study the effect of the composition of the intergranular film (IGF) on anisotropic and isotropic grain growth in alpha-Al2O3. In the simulations, the IGF is formed while in contact with two differently oriented alumina crystals, with the alumina (0001) basal plane on one side and the (110) prism plane on the other. Five different compositions in the IGFs were studied. Results show preferential growth along the [110] of the (110) surface in comparison to growth along the [0001] direction on the (0001) surface for compositions near a Ca/Al ratio of 0.5. Such preferential growth is consistent with anisotropic grain growth in alumina, where platelets form because of faster growth of the prism orientations than the basal orientation. The simulations also show the mechanism by which Ca ions in the IGF inhibit growth on the basal surface. At compositions with high or low Ca/Al ratios, growth along each surface normal is equivalent, indicating isotropic grain growth, although the attachment rates are quite different, which may indicate differences between normal grain growth and abnormal, but isotropic, grain growth. The simulations provide an atomistic view of attachment onto crystal surfaces, affecting grain growth in alumina. PMID- 16471808 TI - FTIR characterization of the reactive interface of cobalt oxide nanoparticles embedded in polymeric matrices. AB - Fourier transform infrared spectroscopy (FTIR) was used as a novel characterization method to determine the properties of the interface that developed when cobalt oxide nanoparticles were self-assembled in a poly(methyl methacrylate) (PMMA) matrix. The method employed the distinct changes that were observed in the infrared spectra of the polymer upon adsorption onto the cobalt oxide nanoparticles, allowing a quantitative determination of the average number of contact points that the average polymer chain formed with the surface of a cobalt oxide nanoparticle of average size. The results obtained with this method compared favorably to those obtained by the coupling of transmission electron microscopy (TEM) experiments with thermogravimetric analysis (TGA). On the basis of both methods, we concluded that the interfacial region created between the cobalt oxide nanoparticles and PMMA is extremely sensitive to the chain length, i.e., the number of anchor points and the density of the polymer layer increase with chain molecular weight. At molecular weights of approximately 250,000, the density of the polymer layer saturates at a value that correspond to that of very thin PMMA films. PMID- 16471810 TI - Redox mediation at 11-mercaptoundecanoic acid self-assembled monolayers on gold. AB - Cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and digital simulation techniques were used to investigate quantitatively the mechanism of electron transfer (ET) through densely packed and well-ordered self assembled monolayers (SAMs) of 11-mercaptoundecanoic acid on gold, either pristine or modified by physically adsorbed glucose oxidase (GOx). In the presence of ferrocenylmethanol (FcMeOH) as a redox mediator, ET kinetics involving either solution-phase hydrophilic redox probes such as [Fe(CN)6]3-/4- or surface-immobilized GOx is greatly accelerated: [Fe(CN)6]3-/4- undergoes diffusion-controlled ET, while the enzymatic electrochemical conversion of glucose to gluconolactone is efficiently sustained by FcMeOH. Analysis of the results, also including the digital simulation of CV and EIS data, showed the prevalence of an ET mechanism according to the so-called membrane model that comprises the permeation of the redox mediator within the SAM and the intermolecular ET to the redox probe located outside the monolayer. The analysis of the catalytic current generated at the GOx/SAM electrode in the presence of glucose and FcMeOH allowed the high surface protein coverage suggested by X-ray photoelectron spectroscopy (XPS) measurements to be confirmed. PMID- 16471812 TI - Reduction of NO2 on ceria surfaces. AB - Cerium dioxide, CeO2, plays an important role in catalysis, due to its ability to store and release oxygen depending on the conditions present in the catalyst environment. To understand the role of ceria in catalytic reactions, we need to study the details of the interaction of ceria surfaces with environmentally sensitive molecules. In this work, we examine the adsorption of the NO2 molecule onto defective (reduced) surfaces of ceria using density functional theory with a correction for on-site Coulomb interactions (DFT+U), which allows for a consistent description of pure and reduced ceria. The interaction of NO2 with defective (111), (110), and (100) surfaces gives an adsorbate-surface structure in which the bond lengths around one Ce(III) ion from the reduced surface shorten, while one N-O bond lengthens. Analysis of the electronic structure and spin density distributions demonstrates that one Ce(III) has been reoxidized to Ce(IV), with the formation of adsorbed NO2-. Finally, we discuss the energetics of the interaction of NO2 with ceria. PMID- 16471811 TI - Homopairing possibilities of the DNA base thymine and the RNA base uracil: an ab initio density functional theory study. AB - All planar homopairings of the DNA base thymine and the RNA base uracil are reported for the first time in this study. Using the idea of binding sites discussed in our previous work (Kelly et al. J. Phys. Chem. B 2005, 109, 11933; J. Phys. Chem. B 2005, 109, 22045) and ab initio density functional theory, we predict and relax 10 thymine and 10 uracil homopairs. The stabilization energies of the homopairs vary from just below zero to -0.82 eV. The results on the pair geometry and energetics are compared with those available in the literature. The collected data on all planar thymine and uracil homopairs can be used to construct the thymine and uracil superstructures seen experimentally on various surfaces. PMID- 16471813 TI - Reactions of hydrazoic acid and trimethylindium on TiO2 rutile (110) surface: a computational study on the formation of the first monolayer InN. AB - This article reports the result of a computational study on the reaction of hydrazoic acid and trimethylindium (TMIn), coadsorbed on TiO2 rutile (110) surface. The adsorption geometries and energies of possible adsorbates including HN3-In(CH3)3(a) and its derivatives, HN3-In(CH3)2(a), N3-In(CH3)2(a), N3 In(CH3)(a), and N-In(a), have been predicted by first-principles calculations based on the density functional theory (DFT) and the pseudopotential method. The mechanisms of these surface reactions have also been explicitly elucidated with the computed potential energy surfaces. Starting from the interaction of three stable HN3 adsorbates, HN3-Ob(a), H(N2)N-Ob(a), and Ti-NN(H)N-Ob(a), where Ob is the bridged O site on the surface, with two stable intermediates from the adsorption and dissociative adsorption of TMIn, (H3C)3In-Ob(a) and (H3C)2In Ob(a)+H3C-Ob(a), InN products can be formed exothermically via four reaction paths following the initial barrierless In-atom association with the N atom directly bonded to H, by CH4 elimination (with approximately 40 kcal/mol barriers), the InN-N bond breaking and the final CH3 elimination or migration (with <20 kcal/mol barriers). These Langmuir-Hinshelwood processes producing the two most stable InN(a) side-on adsorptions confirm that HN3 and TMIn are indeed very efficient precursors for the deposition of InN films on TiO2 nanoparticles. The result of similar calculations for the reactions occurring by the Rideal-Eley mechanism involving HN3(a)+TMIn(g) and HN3(g)+TMIn(a) indicates that they are energetically less favored and produce the less stable InN(a) with end-on configurations. PMID- 16471814 TI - Interlayer expansion and mechanisms of anion sorption of Na-montmorillonite modified by cetylpyridinium chloride: a Monte Carlo study. AB - To study the change of interlayer structure of a Wyoming-type Na-montmorillonite as a result of the replacement of interlayer Na+ ions by cetylpyridinium (CP+) ions, a series of NPT Monte Carlo simulations of the clay mineral with different contents of CP+, Na+, Cl- ions and water in its interlayer space is carried out. In agreement with conclusions from experimental studies, the simulations show that the CP+ ions form monomolecular, bimolecular, and pseudotrimolecular layers with increasing interlayer contents. Calculated potential energies reveal that clay-organic interactions are stronger than organic-organic interactions in CP+ modified montmorillonite, which is in conformity with observations of earlier thermogravimetric experiments. The simulation results indicate that the pseudotrimolecular arrangement of CP+ ions is a prerequisite for the experimentally observed interlayer sorption of inorganic anions. Furthermore, in the interlayer space with a pseudotrimolecular layer, chloride ions favor the formation of pairs with inorganic rather than organic cations. On the basis of these findings and available experimental data, we propose that the interlayer sorption of inorganic anions from the pore space of an organically modified montmorillonite may occur not only in pairs with organic cations, as suggested earlier, but also in pairs with inorganic cations, which represents a so-far unconsidered and maybe more important mechanism of anion sorption on clay minerals. PMID- 16471815 TI - Sum-frequency spectroscopy of a monolayer of zinc arachidate at the solid-solid interface. AB - Infrared-visible sum-frequency spectroscopy has been used to record the vibrational spectrum of a zinc arachidate monolayer at the interface between a sapphire prism and a fused silica lens. Spectra have been recorded for the monolayer deposited on the prism before, during, and after contact, and as a function of increasing pressure. Sum-frequency spectra are reported of the monolayer under sliding contact. The monolayer is found to be resistant to pressure- and shear-induced conformational disorder. However, frequency shifts, drops in peak intensities, and changes in peak intensity ratios have been observed as the monolayer is placed in contact between the prism and the lens. Transfer of monolayer material between the two surfaces is observed and is confirmed by spectra obtained with a monolayer deposited on the surface of the lens rather than the prism. On one face of the sapphire prism, the monolayer reconstructs to a low symmetry layer, probably due to epitaxy. The epitaxial structure disappeared in contact. Existing models for calculating sum-frequency spectra have been extended to include unit cells containing two molecules and torsion about the terminal C-C bond. This model can explain some, but not all, of the experimental observations. PMID- 16471816 TI - Synchrotron photoemission analysis of semiconductor/electrolyte interfaces by the frozen-electrolyte approach: interaction of HCl in 2-propanol with GaAs(100). AB - Perspectives of a new approach for the synchrotron photoemission spectroscopic analysis of chemical processes at solid/liquid interfaces under UHV conditions have been explored. A thin layer of HCl-2-propanol solution was frozen-in on the semiconductor GaAs(100) wafer surface by cooling the substrate to liquid nitrogen temperature after etching off the native oxide layer under N2 atmosphere. Chemical reactions induced in situ by exposure to synchrotron radiation (SR) and by stepwise heating have been monitored. Right after etching and freezing, the surface is covered by gallium chlorides with 1, 2, 3, and 4 Cl ions attached and lattice back-bonded to As atoms, as well as by elemental arsenic As0 and 2 propanol. Exposure to SR at low temperature produces surface As chlorides at the expense of As0. The GaCl3 and GaCl2 emissions diminish while GaCl is enhanced. On the other hand, heating the sample to approximately 130 K just above H2O desorption causes the thermodynamically expected reaction of AsCl3 with the substrate GaAs to form Ga chloride species and As0. Heating the sample to room temperature leaves only As0 on the surface and for gallium the content of all surface chlorides is drastically reduced. By further heating to 400 K elemental arsenic starts to desorb and the Ga chloride surface content is reduced. Using different excitation energies the depth composition of the reaction products has been monitored indicating a tendency of decreasing chlorination numbers and increasing Ga vs As chloride content toward the pristine substrate at each stage of the reaction. PMID- 16471817 TI - Synthesis of, light emission from, and optical power limiting in soluble single walled carbon nanotubes functionalized by disubstituted polyacetylenes. AB - Single-walled carbon nanotubes are covalently functionalized by conjugated polyacetylenes through their cyclization reactions with poly(1-phenyl-1-alkyne) and poly(diphenylacetylene) derivatives carrying azido functional groups at the ends of their alkyl pendants. The resultant polyene nanotube addends are soluble in common solvents, emit intense visible lights and strongly attenuate the power of harsh laser pulses. PMID- 16471818 TI - Examination of spinel and nonspinel structural models for gamma-Al2O3 by DFT and rietveld refinement simulations. AB - Despite the widespread use of gamma-Al2O3, there is still considerable disagreement over the nature of its structure due to both its poor crystallinity and differing preparation techniques during experimentation. Using density functional theory (DFT) calculations and Rietveld simulations and refinement, the structure of three spinel-related models and a recently proposed nonspinel model were studied in reference to synchrotron X-ray powder diffraction (SXPD) patterns. The spinel-based structural models represent the structural features of gamma-Al2O3 better than the nonspinel model. The major failure of the nonspinel model is that the model cannot reproduce the SXPD reflection originating from tetrahedral aluminum. The Rietveld-refined spinel model can accurately reproduce the lattice parameters and other structural features of gamma-Al2O3, and it can generate a consistent diffraction peak at 2theta which lies between the splitting peaks of the experimental pattern that are originated from the disordered tetrahedral aluminum cations. PMID- 16471820 TI - Deprotonation of solvated formic acid: Car-Parrinello and metadynamics simulations. AB - The deprotonation of solvated formic acid was investigated theoretically with ab initio simulations. With the Car-Parrinello method, deprotonation and reprotonation by means of a proton wire were observed. The microscopics of these reactions were analyzed, and reveal the key role played by nearby water molecules in catalyzing the reactions. A constrained molecular dynamics calculation was carried out to estimate the dissociation free energy. Deprotonation of formic acid was further investigated with the recently developed metadynamics method using the formic acid oxygen coordination numbers as the collective variables. The determined free-energy landscape gives barriers similar to that obtained with the constrained free-energy calculation. PMID- 16471821 TI - Prediction of hydrogen hydrate equilibrium by integrating ab initio calculations with statistical thermodynamics. AB - This paper addresses a new calculation approach for the prediction of hydrogen hydrate equilibrium by introducing the concept of a single hydrogen cluster in one cavity. By integrating ab initio calculations with classical statistical thermodynamics, this approach enables the van der Waals model to predict the dissociation pressure of hydrogen hydrates. Compared to hydrates formed by light hydrocarbon gases, structure II (sII) hydrogen hydrates stably encage two and four hydrogen molecules in the small and large cavities, respectively. By treating two hydrogen molecules or four hydrogen molecules as one rigid body cluster, we determine ab initio binding energies between water molecules and hydrogen clusters at the MP2 level with the 6-31++G(2d,2p) basis set. These binding energies will be used to determine the parameters of the Exp-6 potential function from which the smooth cell potential and the Langmuir constant of each cluster are calculated. Then, the dissociation pressure is determined using the Zele-Lee-Holder cell distortion model: 105, 625, and 2000 bar at 150, 200, and 250 K, respectively. PMID- 16471819 TI - Effects of quenched and annealed macromolecular crowding elements on a simple model for signaling in T lymphocytes. AB - Biochemical reactions in cells occur in an environment that is crowded in the sense that various macromolecular species and organelles occupy much of the space. The effects of molecular crowding on biochemical reactions have usually been studied in the past in a spatially homogeneous environment. However, signal transduction in cells is often initiated by the binding of receptors and ligands in two apposed cell membranes, and the pertinent biochemical reactions occur in a spatially inhomogeneous environment. We have studied the effects of crowding on biochemical reactions that involve both membrane proteins and cytosolic molecules by investigating a simplified version of signaling in T lymphocytes using a Monte Carlo algorithm. We find that, if signal transduction occurs on time scales that are slow compared to the motility of the molecules and organelles that constitute the crowding elements, the effects of crowding are qualitatively the same as in a homogeneous three-dimensional (3D) medium. In contrast, if signal transduction occurs on a time scale that is much faster than the time over which the crowding elements move, then the effects of varying the extent of crowding are very different when reactions occur in both 2- and 3D space. We discuss these differences and their origin. Since many signaling reactions are fast, our results may be useful for diverse situations in cell biology. PMID- 16471822 TI - Thermodynamic analysis and thermodynamic efficiency of chemical reactors. AB - In this paper, conditions of minimal dissipation for processes occurring in chemical reactors with a given duration and a given degree of transformation are obtained. It is demonstrated that the derived entropy production can be used to construct the space of realizable (thermodynamically feasible) regimes of a chemical reactor. PMID- 16471823 TI - Effect of structure and thermodynamic stability on the response of lanthanide stannate pyrochlores to ion beam irradiation. AB - The lanthanide stannates, Ln2Sn2O7, Ln=La-Lu and Y, have the isometric pyrochlore structure, A2B2O7, and their structural properties have been refined by Rietveld analysis of powder neutron and synchrotron X-ray diffraction data. In this study, the enthalpies of formation of selected stannate pyrochlores, Ln=La, Nd, Sm, Eu, Dy, and Yb, were measured by high-temperature oxide melt solution calorimetry. Their radiation response was determined by 1 MeV Kr2+ ion irradiation combined with in situ TEM observation over the temperature range of 25 to 1000 K. The enthalpy of formation from binary oxides of stannate pyrochlores became more endothermic (from -145 to -40 kJ/mol) as the size of the lanthanide in the A-site decreases. A more exothermic trend of the enthalpy of formation was observed in stannate pyrochlores with larger lanthanide ions, particularly La, possibly as a result of increased covalency in the Sn-O bond. In contrast to lanthanide titanate pyrochlores, Ln2Ti2O7, that are generally susceptible to radiation induced amorphization and zirconate pyrochlores, Ln2Zr2O7, that are generally resistant to radiation-induced amorphization, the lanthanide stannate pyrochlores show a much greater variation in their response to ion irradiation. La, Nd, and Gd stannates experience the radiation-induced transformation to the aperiodic state, and the critical amorphization temperatures are approximately 960, 700, and 350 K, respectively. Y and Er stannate pyrochlores cannot be amorphized by ion beam irradiation, even at 25 K, and instead disorder to a defect fluorite structure. Comparison of the calorimetric and ion irradiation data for titanate, zirconate, and stannate pyrochlores reveals a strong correlation among subtle changes in crystal structure with changing composition, the energetics of the disordering process, and the temperature above which the material can no longer be amorphized. In summary, as the structure approaches the ideal, ordered pyrochlore structure, radiation-induced amorphization is more easily attained. This is consistent with an increasingly exothermic trend in the enthalpies of formation of pyrochlores from the oxides, that is, the greater the thermochemical stability of the pyrochlore structure, the more likely it will be amorphized upon radiation damage rather than recover to a disordered fluorite structure. PMID- 16471824 TI - Molecular dynamics study of the properties of capsaicin in an 1-octanol/water system. AB - Molecular dynamics simulations were performed to study the behavior of capsaicin in an 1-octanol/water system at 298 K and 1 bar. Capsaicin is the pungent chemical found in chili pepper that stimulates our sensory system resulting in a burning, pain sensation. In the first step toward investigating the activity of capsaicin, we have used two molecular representations for capsaicin based on the OPLS force field: all-atom and united-atom models. The octanol/water mixture was selected as a model system to determine the hydrophobic and hydrophilic properties of capsaicin by analyzing equilibrium, structural, and dynamic properties from the simulations. Our simulations showed that capsaicin preferentially partitions to the octanol phase, with its hydrocarbon segment oriented with that in octanol, while the polar part remains exposed to the aqueous phase. The simulations with the all-atom and united-atom models yielded similar results. PMID- 16471825 TI - Model of an asymmetric DPPC/DPPS membrane: effect of asymmetry on the lipid properties. A molecular dynamics simulation study. AB - The study of asymmetric lipid bilayers is of a crucial importance due to the great number of biological process in which they are involved such as exocytosis, intracellular fusion processes, phospholipid-protein interactions, and signal transduction pathway. In addition, the loss of this asymmetry is a hallmark of the early stages of apoptosis. In this regard, a model of an asymmetric lipid bilayer composed of DPPC and DPPS was simulated by molecular dynamics simulation. Thus, the asymmetric membrane was modeled by 264 lipids, of which 48 corresponded to DPPS- randomly distributed in the same leaflet with 96 DPPC. In the other leaflet, 120 DPPC were placed without DPPS-. Due to the presence of a net charge of -1 for the DPPS- in physiological conditions, 48 Na+ were introduced into the system to balance the charge. To ascertain whether the presence of the DPPS- in only one of the two leaflets perturbs the properties of the DPPC in the other leaflet composed only of DPPC, different properties were studied, such as the atomic density of the different components across the membrane, the electrostatic potential across the membrane, the translational diffusion of DPPC and DPPS, the deuterium order parameters, lipid hydration, and lipid-lipid charge bridges. Thus, we obtained that certain properties such as the surface area lipid molecule, lipid head orientation, order parameter, translational diffusion coefficient, or lipid hydration of DPPC in the leaflet without DPPS remain unperturbed by the presence of DPPS in the other leaflet, compared with a DPPC bilayer. On the other hand, in the leaflet containing DPPS, some of the DPPC properties were strongly affected by the presence of DPPS such as the order parameter or electrostatic potential. PMID- 16471826 TI - Ruthenium(II) complex of Hbopip: synthesis, characterization, pH-induced luminescence "off-on-off" switch, and avid binding to DNA. AB - A novel ruthenium(II) complex of [Ru(bpy)2(Hbopip)](ClO4)2 (in which bpy=2,2' bipyridine, Hbopip=2-(4-benzoxazolyl)phenylimidazo[4,5-f][1,10]phenanthroline) was synthesized and characterized. The spectrophotometric pH titrations of the complex showed that it acted as a pH-induced luminescence "off-on-off" switch: a luminescence off-on switch with a luminescence enhancement factor of IpH=3.0/IpH=1.0=20 occurring over a narrow pH range of 1.00-3.00 plus a luminescence on-off switch with a luminescence enhancement factor of 3 over a pH range of 3.20-9.40. The excited-state ionization constant of the complex derived, pKa1*=3.06, is 1.36 pKa units greater than the ground-state pKa1=1.70, and pKa2*=5.01 and pKa3*=8.22 are comparable to the ground-state pKa2=5.23 and pKa3=8.22, respectively. The complex avidly bound to calf thymus DNA with a large binding constant of (1.2+/-0.3)x10(7) M-1 in buffered 50 mM NaCl, as evidenced by UV-vis and luminescence titrations, steady-state emission quenching by [Fe(CN)6]4 , DNA competitive binding with ethidium bromide, viscosity measurements, and DNA melting experiments. PMID- 16471827 TI - Peracetylated bovine carbonic anhydrase (BCA-Ac18) is kinetically more stable than native BCA to sodium dodecyl sulfate. AB - Bovine carbonic anhydrase (BCA) and its derivative with all lysine groups acetylated (BCA-Ac18) have different stabilities toward denaturation by sodium dodecyl sulfate (SDS). This difference is kinetic: BCA-Ac18 denatures more slowly than BCA by several orders of magnitude over concentrations of SDS ranging from 2.5 to 10 mM. The rates of renaturation of BCA-Ac18 are greater than those of BCA, when these proteins are allowed to refold from a denatured state ([SDS]=10 mM) to a folded state ([SDS]=0.1 to 1.5 mM). On renaturation, the yields of the correctly folded protein (either BCA or BCA-Ac18) decrease with increasing concentration of SDS. At intermediate concentrations of SDS (from 0.7 to 2 mM for BCA, and from 1.5 to 2 mM for BCA-Ac18), both unfolding and refolding of the proteins are too slow to be observed; an alternative process-probably aggregation competes with refolding of the denatured proteins at those intermediate concentrations. Because it is experimentally impractical to prove equilibrium, it is not possible to establish whether there is a difference in the thermodynamics of unfolding/refolding between BCA and BCA-Ac18. PMID- 16471828 TI - Kinetics of membrane raft formation: fatty acid domains in stratum corneum lipid models. AB - The major barrier to permeability in skin resides in the outermost layer of the epidermis, the stratum corneum (SC). The major SC lipid components are ceramides, free fatty acids, and cholesterol. Ternary mixtures containing these constituents are widely used for physicochemical characterization of the barrier. Prior X-ray diffraction and IR spectroscopy studies have revealed the existence of ordered lipid chains packed in orthorhombic subcells. To monitor the kinetics of formation of regions rich in fatty acids, the current study utilizes a modification of the method (J. Phys. Chem. 1992, 96, 10008) developed to monitor component demixing in n-alkane mixtures. The approach is based on changes in the scissoring or rocking mode contours in the IR spectra of (orthorhombically packed) ordered chains. In the current study, equimolar mixtures of ceramides (either non-hydroxy fatty acid sphingosine ceramide or alpha-hydroxy fatty acid sphingosine ceramide) with chain perdeuterated fatty acids (either palmitic or stearic acid) and cholesterol reveal a time evolution of the scissoring contour of the deuterated fatty acid chains following quenching from relatively high temperatures where random mixing occurs. Segregation of domains enriched in the fatty acid component is observed. The kinetics of segregation are sensitive to the quenching temperature and to the chemical composition of the mixture. The kinetic regimes are conveniently catalogued with a power law of the form P=Ktalpha where P is a (measured) property related to domain composition. The time scales for demixing in these experiments are similar to times observed in several studies that have tracked the restoration of the in vivo permeability barrier following nonthermal challenges to SC integrity. Further evidence for the physiological importance of the current measurements is the detection of these phases in native SC. The current work constitutes the first direct, structure based determination of the kinetics of barrier formation in relevant skin lipid barrier models. PMID- 16471829 TI - A model for DNA detection by metal-enhanced fluorescence from immobilized silver nanoparticles on solid substrate. AB - N-(2-Mercaptopropionyl)glycine (tiopronin)-coated silver nanoparticles (average diameter of metal core=5 nm) were prepared by a modified Brust method. Tiopronin ligands were partially displaced by thiolate single-stranded oligonucleotides via ligand exchange. These particles were immobilized onto a solid substrate through hybridization with target oligonucleotides in a layer-by-layer approach. The dye labeled complementary oligonucleotides were bound to the particle layers through hybridization. Fluorescence intensity was enhanced with a simultaneous increase of plasmon absorbance from accumulated particles. A steady state was shown at the 10th particle layer and then the fluorescence enhancement showed a plateau. This result reveals that increasing the particle layer contributes to fluorescence enhancement. This novel method was used to detect DNA hybridization through both absorbance and emission spectral changes. PMID- 16471830 TI - Degradation of protein in nanoplasma generated around gold nanoparticles in solution by laser irradiation. AB - We developed a method of protein degradation in an aqueous solution containing gold nanoparticles by irradiation of a pulse laser. In the present study, lysozyme was used as an example. Lysozyme degradation proceeded most efficiently when a pH of the solution was adjusted so that it was at the isoelectric point. The scheme of the lysozyme degradation is as follows: (1) Lysozyme molecules in the solution are neutralized and adsorbed on the gold nanoparticles with its pH value adjusted at the isoelectric point, (2) nanoplasma is generated in the close vicinity of a gold nanoparticle which is excited by an intense 532-nm laser, (3) lysozyme molecules in the nanoplasma are degraded into small fragments. Lysozyme degradation does not proceed efficiently at a pH value deviated from the isoelectric point because the lysozyme molecules are dissolved uniformly so that only a small portion of the lysozyme molecules are located in the vicinity of gold nanoparticles which create the nanoplasma. PMID- 16471831 TI - Spectroscopic properties of porphyrin-like photosensitizers: insights from theory. AB - Electronic absorption spectra of six porphyrin-like photosensitizers, porphyrin, chlorin, bacteriochlorin, pheophytin a, porphyrazin, and texaphyrin, have been calculated within the time-dependent DFT framework (TDDFT) in conjunction with the PBE0 hybrid functional. Energetic and orbital aspects are discussed by comparing systems together so as to assess the best molecules for photodynamic therapy applications. Excitation energies and oscillator strengths are found to be in good agreement with both experimental data and previous theoretical works. In particular, whereas significant discrepancies (0.3 eV) appear for Qx bands, results become more reliable as wavelengths decrease. To elucidate the effect of the local environment, we have taken into account solvation either with explicit water molecules interacting via hydrogen bonds with the system or with a continuum model (C-PCM). The supramolecular approach does not affect spectra, while using C-PCM improves Qx and B band values and strengthens intensities significantly. In both gaseous and aqueous phases, texaphyrin, pheophytin a, and bacteriochlorin Qx bands are found in the 600-800 nm range as expected by experimental works. These data are particularly interesting in the perspective of systematic studies of other photosensitizers and should make experimentalists' works easier. PMID- 16471832 TI - Conformational properties of and a reorientation triggered by sugar-water vibrational resonance in the hydroxymethyl group in hydrated beta-glucopyranose. AB - In this paper, we discuss the conformational properties of the hydroxymethyl group of beta-glucopyranose in aqueous solution and its reorientation mechanism. First, using the values for the hydroxymethyl torsion (O5-C5-C6-O6) angle obtained by our ab initio simulations, we reestimate the experimental ratio of the hydroxymethyl rotamer populations. The reestimated ratio is found to be in agreement with those previously reported in several computational studies, which probably partly explains the discrepancies between theoretical and experimental studies that have been discussed in the literature. Second, our time-frequency analysis on a reorientation in the hydroxymethyl group in an ab initio molecular dynamics trajectory suggests that, before the reorientation, the O6-H6 stretching mode is vibrationally coupled with a proton-accepting first-hydration-shell water molecule, whereas the C6-O6 stretching mode is vibrationally coupled with a proton-donating one. The amount of the total vibrational energy induced by these vibrational couplings is estimated to be comparable to typical values for the potential barriers between hydroxymethyl rotamers. To elucidate the vibrational couplings, we investigate the hydrogen-bonding properties around the hydroxymethyl group during the pretransition period. The implications, validity, and limitation of a possible reorientation mechanism based on these findings are also discussed. PMID- 16471833 TI - Evidence for non-two-state kinetics in the nucleocapsid protein chaperoned opening of DNA hairpins. AB - In HIV-1 reverse transcription, the nucleocapsid protein, NC, induces secondary structure fluctuations in specific DNA and RNA hairpins. Time-resolved single molecule fluorescence resonance energy transfer was used to study NC chaperoned opening of DNA hairpins over a broader range of conditions and in more depth than in previous studies. The experiments reveal a complex mechanism for secondary structure fluctuations with dynamic processes occurring over a wide time range, i.e., approximately 5 to >250 ms and with the involvement of long-lived intermediates. The dynamic role of DNA loop regions and NC binding/dissociation events are discussed. PMID- 16471834 TI - Fluid self-diffusion in Scots pine sapwood tracheid cells. AB - The self-diffusion coefficients of water and toluene in Scots pine sapwood was measured using low field pulsed field gradient nuclear magnetic resonance (PFG NMR). Wood chips of 8 mm diameter were saturated with the respective liquids, and liquid self-diffusion was then traced in one dimension orthogonal to the tracheid cell walls in the wood's radial direction. The experimental echo attenuation curves were exponential, and characteristic self-diffusion coefficients were produced for diffusion times spanning from very short times to times on the order of magnitude of seconds. Observed self-diffusion coefficients were decaying asymptotically as a function of diffusion time, an effect which was ascribed to the cell walls' restriction on confined liquid diffusion. The observed self diffusion behavior in Scots pine sapwood was compared to self-diffusion coefficients obtained from simulations of diffusion in a square. Principles of molecular displacements in confined geometries were used for elucidating the wood's cellular structure from the observed diffusion coefficients. The results were compared with a mathematical model for diffusion between parallel planes. PMID- 16471835 TI - Freezing a single distal motion in dihydrofolate reductase. AB - Constraining a single motion between distal residues separated by approximately 28 A in hybrid quantum/classical molecular dynamics simulations is found to increase the free energy barrier for hydride transfer in dihydrofolate reductase by approximately 3 kcal/mol. Our analysis indicates that a single distal constraint alters equilibrium motions throughout the enzyme on a wide range of time scales. This alteration of the conformational sampling of the entire system is sufficient to significantly increase the free energy barrier and decrease the rate of hydride transfer. Despite the changes in conformational sampling introduced by the constraint, the system assumes a similar transition state conformation with a donor-acceptor distance of approximately 2.72 A to enable the hydride transfer reaction. The modified thermal sampling leads to a substantial increase in the average donor-acceptor distance for the reactant state, however, thereby decreasing the probability of sampling the transition state conformations with the shorter distances required for hydride transfer. These simulations indicate that fast thermal fluctuations of the enzyme, substrate, and cofactor lead to conformational sampling of configurations that facilitate hydride transfer. The fast thermal motions are in equilibrium as the reaction progresses along the collective reaction coordinate, and the overall average equilibrium conformational changes occur on the slower time scale measured experimentally. Recent single molecule experiments suggest that at least some of these thermally averaged equilibrium conformational changes occur on the millisecond time scale of the hydride transfer reaction. Thus, introducing a constraint that modifies the conformational sampling of an enzyme could significantly impact its catalytic activity. PMID- 16471836 TI - Resorcarene-based receptor: versatile behavior in its interaction with heavy and soft metal cations. AB - Standard solution Gibbs energies, DeltasG degrees, of the resorcarene-based receptor 5,11,17,23-ethylthiomethylated calix[4]resorcarene, (characterized by 1H NMR and X-ray diffraction studies) in its monomeric state (established through partition experiments) in various solvents are for the first time reported in the area of resorcarene chemistry. Transfer Gibbs energies of from hexane (reference solvent) to other medium are calculated. Agreement between DeltatG degrees (referred to the pure solvents) and standard partition Gibbs energies, DeltapG degrees (solvent mutually saturated) is found. Cation-ligand interactions were investigated through 1H NMR (CD3CN and CD3OD) and conductometric titrations in acetonitrile and methanol. 1H NMR data revealed the sites of interaction of with the metal cation. The composition of the metal-ion complexes (Ag+ and Pb2+ in acetonitrile and Ag+ and Cu2+ in methanol) was established through conductometric titrations. Thus, complexes of 1:1 stoichiometry were formed between and Ag+ and Pb2+ in acetonitrile and Cu2+ in methanol. However, in moving from acetonitrile to methanol, the composition of the silver complex was altered. Thus, two metal cations are hosted by a unit of the ligand. As far as Cu2+ and in acetonitrile is concerned, conductance data suggest that metalates are formed in which up to four units of Cu2+ are taken up per unit of resorcarene. The contrasting behavior of with Cu2+ in acetonitrile relative to methanol is discussed. As far as mercury (II) is concerned, the unusual jump in conductance observed in the titration of Hg2+ with in acetonitrile and methanol after the formation of a multicharged complex (undefined composition) is attributed to the presence of highly charged smaller units (higher mobility) resulting from the departure of pendant arms from the resorcarene backbone. Isolation of these species followed by X-ray diffraction studies corroborated this statement. The thermodynamic characterization of metal-ion complexes of Ag+ and Pb2+ in acetonitrile and Cu2+ and Ag+ in methanol is reported. Final conclusions are given. PMID- 16471837 TI - Kinetic effects of alcohol addition on the anodic behavior of silicon in acid fluoride media. AB - The anodic dissolution of silicon in acid solutions of ammonium fluoride is investigated in aqueous and water-alcohol media by rotating disk voltammetry and impedance spectroscopy. The voltammograms recorded in water-alcohol media show, in comparison to water, the following effects: an increase of the dissolution currents measured at pH>2 and an opposite effect at pH<2; a distortion of the curve, with a relative increase of the characteristic currents in the region of low applied potentials, indicating easier dissolution of the "wet" oxide forming under those conditions; a shift of the maximum of the current-pH curves from about pH 3 in water to about pH 4 in 50% ethanol v/v. Analysis of impedance data at pH values 4-5 in water and water-ethanol media show similar dependence on potential for the parameters Chf (high-frequency capacitance) and RhfI (Rhf is a high-frequency resistance and I is the steady state current), suggesting the formation of oxide films with similar properties. The marked differences of anodic behavior in the two types of media are discussed and explained in terms of the effects of alcohol addition on the speciation equilibria of hydrofluoric acid and on the kinetic rate constants of dissolution of the surface oxide layer. PMID- 16471839 TI - Probing the Raman scattering tensors of individual molecules. AB - Single-molecule experiments provide new views into the mechanisms behind surface enhanced Raman scattering. It was shown previously that spectra of individual rhodamine 6G molecules adsorbed on silver nanocrystal aggregates present stronger fluctuations in two low-frequency bending modes, at 614 and 773 cm(-1). Here we use polarization spectroscopy to show that these bands are enhanced by a resonant process whose transition dipole is rotated by 15+/-10 degrees with respect to the molecular transition dipole. We also show that the polarization function remains stable over the whole time scale of a measurement, indicating that molecular reorientation with respect to the surface is unlikely. Together these findings provide further support to the involvement of a charge-transfer resonance in the enhancement of the low-frequency bands and allow us to suggest a model for the orientation of rhodamine 6G molecules at Raman hot spots. PMID- 16471840 TI - Variability of conductance in molecular junctions. AB - The conductance of molecular junctions, formed by breaking gold point contacts dressed with various thiol functionalized organic molecules, is measured at 293 K and at 30 K. In the presence of molecules, individual conductance traces measured as a function of increasing gold electrode displacement show clear steps below the quantum conductance steps of the gold contact. These steps are distributed over a wide range of molecule-dependent conductance values. Histograms constructed from all conductance traces therefore do not show clear peaks either at room or low temperatures. Filtering of the data sets by an objective automated procedure only marginally improves the visibility of such features. We conclude that the geometrical junction to junction variations dominate the conductance measurements. PMID- 16471841 TI - Unexpected formation of higher polythionates in the oxidation of thiosulfate by hypochlorous acid in a slightly acidic medium. AB - It has been clearly shown that not only tetrathionate but also pentathionate or even higher polythionates is formed in the oxidation of thiosulfate by hypochlorous acid in a slightly acidic medium. In thiosulfate excess, the absorbance-time curves registered at 250 nm may go through a maximum followed by a minimum, suggesting the presence of a short-lived absorbing intermediate proposed to be S(2)O(3)OCl(3-). Matrix rank analysis (MRA) studies have revealed that altogether five independent absorbing species are present in the wavelength range 240-400 nm. A kinetic model is suggested to take all of the experimental observations into account. PMID- 16471842 TI - Composite phospholipid-calcium carbonate microparticles: influence of anionic phospholipids on the crystallization of calcium carbonate. AB - The synthesis and characterization of calcium carbonate microparticles by reaction of calcium chloride and ammonium bicarbonate in the presence of negatively charged phospholipid mixtures of negative and zwitterionic phospholipids has been reported. Negatively charged phospholipids influence the crystal morphology of calcium carbonate and induce the formation of thermodynamically less stable veterite polymorph as opposed to calcite polymorph. The phospholipids are entrapped in the calcium carbonate microparticles during the crystallization process, with a uniform distribution of phospholipids in the interior of the microparticles. This phenomenon was exploited to encapsulate a model hydrophobic fluorophore, the tris(4,7-diphenyl-1,10 phenanthroline)ruthenium(II) dichloride complex, to simulate encapsulation of hydrophobic drug molecules. Thermogravimetric analysis reveals that, in these microparticles, the calcium carbonate and the phospholipid exhibit strong interactions. PMID- 16471843 TI - Coating single-walled carbon nanotubes with phospholipids. AB - Single-walled carbon nanotubes (SWNTs), being hydrophobic by nature, aggregate in water to form large bundles. However, isolated SWNTs possess unique physical and chemical properties that are desirable for sensing and biological applications. Conventionally isolated SWNTs can be obtained by wrapping the tubes with biopolymers or surfactants. The binding modes proposed for these solubilization schemes, however, are less than comprehensive. Here we characterize the efficacies of solubilizing SWNTs through various types of phospholipids and other amphiphilic surfactants. Specifically, we demonstrate that lysophospholipids, or single-chained phospholipids offer unprecedented solubility for SWNTs, while double-chained phospholipids are ineffective in rendering SWNTs soluble. Using transmission electron microscopy (TEM) we show that lysophospholipids wrap SWNTs as striations whose size and regularity are affected by the polarity of the lysophospholipids. We further show that wrapping is only observed when SWNTs are in the lipid phase and not the vacuum phase, suggesting that the environment has a pertinent role in the binding process. Our findings shed light on the debate over the binding mechanism of amphiphilic polymers and cylindrical nanostructures and have implications on the design of novel supramolecular complexes and nanodevices. PMID- 16471844 TI - From subtle to substantial: role of metal ions on pi-pi interactions. AB - Quantum chemistry calculations reveal that the subtle pi-pi interactions, usually in the range 2-4 kcal/mol, will become substantially significant, from 6 to 17 kcal/mol, in the presence of metal ion. The metal ions have higher affinity toward a pi-pi dimer compared to a single pi-moiety. Considering the widespread occurrence of cation-pi-pi motifs in chemistry and biology, as evident from the database analysis, we propose that the two key noncovalent forces, which govern the macromolecular structure, cation-pi and pi-pi, work in concert. PMID- 16471846 TI - Solvothermal synthesis of monodisperse PbSe nanocrystals. AB - Size-controllable monodisperse PbSe and PbSe/PbS nanocrystals (NCs) have been successfully synthesized with a solvothermal method. Octadecylamine (ODA) molecules were found effective in organizing the nanocrystals to form an ordered monolayer. It is expected that these narrow-band-gap semiconductor NCs with tunable size would have potential applications in near- and mid-IR telecommunication laser sources, electroluminescence, and solar cell materials. PMID- 16471845 TI - Principles and implementations of dissipative (dynamic) self-assembly. AB - Dynamic self-assembly (DySA) processes occurring outside of thermodynamic equilibrium underlie many forms of adaptive and intelligent behaviors in natural systems. Relatively little, however, is known about the principles that govern DySA and the ways in which it can be extended to artificial ensembles. This article discusses recent advances in both the theory and the practice of nonequilibrium self-assembly. It is argued that a union of ideas from thermodynamics and dynamic systems' theory can provide a general description of DySA. In parallel, heuristic design rules can be used to construct DySA systems of increasing complexities based on a variety of suitable interactions/potentials on length scales from nanoscopic to macroscopic. Applications of these rules to magnetohydrodynamic DySA are also discussed. PMID- 16471847 TI - Evaluation of force fields for molecular simulation of polyhedral oligomeric silsesquioxanes. AB - Polyhedral oligomeric silsesquioxanes (POSS) are nanometer-size molecules suitable for the production of organic-inorganic nanocomposite materials. These organic-inorganic nano-building blocks show promise for enabling the production of polymeric materials of exceptional mechanical properties as well as novel composite materials. While the experimental studies of these materials have rapidly evolved in the past decade, their theoretical investigation is still in its infancy. Toward the validation of force fields for the molecular simulation of POSS-containing systems, we present the charge-transfer reactive (CTR) force field for the molecular simulation of polyhedral oligomeric silsesquioxane (POSS) molecules and compare the ability of this, and several force fields taken from the literature, to predict the thermophysical properties of POSS-containing systems. The literature force fields compared include the universal force field (UFF) and the COMPASS and Hybrid-COMPASS force fields. Predictions from molecular dynamics simulations of the structural parameters (unit cell vectors), melting temperature, and FT-IR spectra of crystals of POSS monomers are presented. The POSS monomers investigated are octahydride, octamethyl, and octapropyl POSS. Predicted quantities are compared to experimental results where available and provide molecular-level physical insight into several aspects of the behavior of POSS molecules. While all the force fields tested perform reasonably well, our results indicate that the Hybrid-COMPASS and CTR force fields predict structural properties that are in good agreement with experimental data. PMID- 16471848 TI - On the volume-dependence of the index of refraction from the viewpoint of the complex dielectric function and the Kramers-Kronig relation. AB - How indices of refraction n(omega) of insulating solids are affected by the volume dilution of an optical entity and the mixing of different, noninteracting simple solid components was examined on the basis of the dielectric function epsilon(1)(omega) + iepsilon(2)(omega). For closely related insulating solids with an identical composition and the formula unit volume V, the relation [epsilon(1)(omega) - 1]V = constant was found by combining the relation epsilon(2)(omega)V = constant with the Kramers-Kronig relation. This relation becomes [n(2)(omega) - 1]V = constant for the index of refraction n(omega) determined for the incident light with energy less than the band gap (i.e., h omega < E(g)). For a narrow range of change in the formula unit volume, the latter relation is well approximated by a linear relation between n and 1/V. PMID- 16471849 TI - Nanostructured oxide-based powders: investigation of the growth mode of the CeO2 clusters on the YSZ surface. AB - CeO(2)/YSZ nanocomposite powders, characterized by increasing Ce/Zr atomic ratio, were obtained by depositing, by wet impregnation, different amounts of CeO(2) on the yttria-stabilized zirconia (YSZ) surface. These powders were characterized by means of X-ray photoelectron spectroscopy, transmission electron microscopy, energy dispersive spectroscopy, and X-ray diffraction. Experimental results allow us to obtain interesting information concerning the growth mode, the morphology, and the dimensions of the CeO(2) clusters on the YSZ supporting surface. A 3-D growing mechanism was observed for the CeO(2) nanoparticles. With increasing Ce/Zr atomic ratio the CeO(2) clusters become more and more spherical. Moreover, XPS data also show the presence of Ce(III) and Ce(IV) ions at the interface supported/supporting oxides. PMID- 16471850 TI - Electrohydrodynamic atomization: an approach to growing continuous self supporting polymeric fibers. AB - The investigation presented in this paper illustrates a technique for growing in situ polymerized networks, forming scaffold-like structures usually formed by means of electrospinning. The technique of jet atomization employing electrohydrodynamics is a manifestation of electrospinning. However, we show for the first time that using this technique where individual droplets are generated, a continuous self-supporting submicrometer web-like structure can be grown whereby fragments of the structure are delivered in the droplets and polymerize on the surface of the growing structure via polycondensation. The development of these growing fibers into web structures is a direct result of the processing route together with the excellent tailor-made cross-linking nature of the resin. An operational map is generated to identify a parametric space in which the stable cone-jet mode of electrohydrodynamic atomization prevails for generating the finest droplets. A statistical analysis on the formed fibers for a given time and electrospray condition is presented together with optical micrographs of the structure, which concludes the discussion in this paper. PMID- 16471851 TI - True nanocable assemblies with insulating BN nanotube sheaths and conducting Cu nanowire cores. AB - Nanocable models comprised of BN nanotubes filled with close-packed Cu nanowires were investigated by gradient-corrected density functional theory (DFT) computations. The optimal distance between the sidewall of BN nanotubes and the atoms in a copper nanowire is about 0.35 nm, with a weak insertion energy (ca. 0.04 eV per Cu atom). Hence, such nanocables are assembled by weaker van der Waals (vdW) forces, rather than by chemical bonding interactions. The electronic band structures of the BN/Cu hybrid systems are superposition of those of the separate components, the BN nanotubes, and the Cu nanowires. Since charge density analyses show that the conduction electrons are distributed only on the copper atoms, charge transport will occur only in these inner nanowires, which are effectively insulated by the outer BN nanotubes. On the basis of these computational results, BN/Cu hybrid structures should be ideal nanocables. PMID- 16471852 TI - Enantioselective photooxidation of a sulfide by a chiral ruthenium(II) complex immobilized on a montmorillonite clay surface: the role of weak interactions in asymmetric induction. AB - The present work pursued a possibility that enantioselectivity was achieved through weak intermolecular interactions between a catalyst and a substrate. For that purpose, we studied the photooxidation of alpha-ethylbenzyl phenyl sulfide catalyzed by a polypyridyl ruthenium(II) complex as a chiral photosensitizer. No covalent bonding was formed between a catalyst and a substrate, because the complexes used ([Ru(phen)(3)](2+) or [Ru(bpy(3))(2+)]) were coordinatively saturated. Enantiomer excess (ee) was attained to be 30% when a chiral photosensitizer was immobilized on montmorillonite clay. It was even improved to 43% in the presence of an additional chiral auxiliary, dibenzoyl-D(+)-tartaric acid. Notably, no enantioselectivity was achieved when the reaction took place in homogeneous solutions. The ab initio calculations were performed on the stability of an associate composed of a catalyst (metal complex) and a product (sulfoxide) to obtain a clue to reaction mechanisms. The calculations suggest that chiral discrimination is achieved even through noncovalent interactions between a substrate and a chiral sensitizer when the attacking direction by a substrate toward a catalyst is limited sterically on a solid surface. PMID- 16471853 TI - Influence of copolymer configuration on the phase behavior of ternary blends. AB - The influence of copolymer configuration on the phase behavior of various ternary polymer blends containing a crystallizable polyester, a noncrystallizable polyether, and an acrylic random copolymer of different chain configuration was investigated. In these ternary blends, the acrylic random copolymer is typically added to control rheological properties at elevated temperatures. In fact, the acrylic random copolymers composed of various compositions of MMA and nBMA were found to have different miscibility with polyester as well as polyether, leading to substantially different phase behavior of ternary blends. Remarkable temperature dependence was also found. The mean-field Flory-Huggins theory for the free energy of mixing, extended to ternary polymer blends, was adopted for predicting phase diagrams where the exact spinodal and binodal boundaries could be calculated. Phase diagrams of ternary blends, predicted by the Flory-Huggins formulations and related calculations, were in good agreement with experimental phase diagrams. The differences observed in the rheological processes of various ternary blends with different acrylic copolymers were directly related to changes in miscibility, associated phase behavior, and chain configuration. PMID- 16471854 TI - Laser-assisted synthesis of Au-Ag alloy nanoparticles in solution. AB - By using laser-induced heating, we prepared Au-Ag nanoalloys via three different procedures: (i) mixture of Au nanoparticles and Ag(+) ions irradiated by a 532 nm laser, (ii) mixture of Au and Ag nanoparticles irradiated by a 532 nm laser, and (iii) mixture of Au and Ag nanoparticles irradiated by a 355 nm laser. Procedure i is advantageous for the production of spherical alloy nanoparticles; in procedures ii and iii, nanoalloys with a sintered structure have been obtained. The morphology of the obtained nanoalloys depends not only on the laser wavelength but also on the concentration of nanoparticles in the initial mixture. When the total concentration of Ag and Au nanoparticles in the mixture is increased, large-scale interlinked networks have been observed upon laser irradiation. It is expected that this selective heating strategy can be extended to prepare other bi- or multi-metallic nanoalloys. PMID- 16471855 TI - Theoretical study of the influence of monomer excess on the structure and properties of polyaniline oligomers. AB - Polyaniline is among the most intensely investigated polymers because of its exceptional properties affording its current and potential applications. The structure and energy spectra of isolated oligomers and infinite chains in different oxidation states and degrees of protonation have been discussed at length from experimental and theoretical perspectives. The reaction environment effect, however, has received less attention and, particularly, the influence of monomer excess has been completely neglected in theoretical studies. Experimental measurements show that residual aniline is always detected in emeraldine samples obtained at low pH. Upon addition of oxidant to emeraldine PANI samples, post polymerization due to the presence of excess monomers occurs. This is an indication of the formation of aniline-PANI complexes in the reaction medium. The presence of aniline monomers should affect the PANI chain arrangement and optical/conducting characteristics. Therefore, model clusters of aniline with neutral or singly protonated emeraldine tetramers in explicit water medium and periodic boundary conditions are addressed in this paper using a Monte Carlo/AMBER96/AM1 computational protocol to simulate the absorption spectra. The monomer impact on the structure, energy characteristics, and UV/vis spectra of the polymer are discussed. PMID- 16471856 TI - Mechanism of oxygen electroreduction on gold surfaces in basic media. AB - The mechanism of the electroreduction of oxygen on Au surfaces in basic media is examined using surface-enhanced Raman scattering (SERS) measurements and density functional theory (DFT) calculations. The spectroscopy reveals superoxide species as a reduction intermediate throughout the oxygen electroreduction, while no peroxide is detected. The spectroscopy also shows the presence of superoxide after the addition of hydrogen peroxide. The calculations show no effect of OH addition to the Au(100) surface with regard to O-O length. These results suggest that the four-electron reduction of O(2) on Au(100) in base arises from a disproportionation mechanism which is enhanced on Au(100) relative to the other two low Miller index faces of Au. PMID- 16471857 TI - Partial oxidation of propylene to propylene oxide over a neutral gold trimer in the gas phase: a density functional theory study. AB - We report a B3LYP study of a novel mechanism for propylene epoxidation using H(2) and O(2) on a neutral Au(3) cluster, including full thermodynamics and pre exponential factors. A side-on O(2) adsorption on Au(3) is followed by dissociative addition of H(2) across one of the Au-O bonds (DeltaE(act) = 2.2 kcal/mol), forming a hydroperoxy intermediate (OOH) and a lone H atom situated on the Au(3) cluster. The more electrophilic O atom (proximal to the Au) of the Au OOH group attacks the C=C of an approaching propylene to form propylene oxide (PO) with an activation barrier of 19.6 kcal/mol. We predict the PO desorption energy from the Au(3) cluster with residual OH and H to be 11.5 kcal/mol. The catalytic cycle can be closed in two different ways. In the first subpathway, OH and H, hosted by the same terminal Au atom, combine to form water (DeltaE(act) = 26.5 kcal/mol). We attribute rather a high activation barrier of this step to the breaking of the partial bond between the H atom and the central Au atom in the transition state. Upon water desorption (DeltaE(des) = 9.9 kcal/mol), the Au(3) is regenerated (closure). In the second subpathway, H(2) is added across the Au OH bond to form water and another Au-H bond (DeltaE(act) = 22.6 kcal/mol). Water spontaneously desorbs to form an obtuse angle Au(3) dihydride, with one H atom on the terminal Au atom and the other bridging the same terminal Au atom and the central Au atom. A slightly activated rearrangement to a symmetric triangular Au(3) intermediate with two equivalent Au-H bonds, addition of O(2) into the Au-H bond, and rotation reforms the hydroperoxy intermediate in the main cycle. On the basis of the DeltaG(act), which contains contribution from both pre-exponetial factor and activation energy, we identify the propylene epoxidation step as the actual rate-determining step (RDS) in both the pathways. The activation barrier of the RDS (epoxidation step: DeltaE(act) = 19.6 kcal/mol) is in the same range as that in the published computationally investigated olefin epoxidation mechanisms involving Ti sites (without Au involved) indicating that isolated Au clusters and possibly Au clusters on non-Ti supports can be active for gas-phase partial oxidation, even though cooperative mechanisms involving Au clusters/Ti based-supports may be favored. PMID- 16471858 TI - The influence of flexible segments on liquid crystalline properties in terms of solubility parameters. An attempt at quantitative interpretation. AB - The phase behavior of some rodlike block molecules has been reviewed with reference to the polarity of constituent segments. It was found that the ability of the mesophase formation is connected with differences in polar character between the flexible chains and rigid cores. Thus the polar poly(oxyethylene) group connected with the polar rigid core reduces mesophase stability but is advantageous when put together with some apolar building blocks. An attempt at quantitative estimation of the incompatibilities of different parts of molecules by means of Hansen solubility parameters delta and Flory interaction parameters chi has also been made. On the basis of chi parameters the Gibbs free energies of mixing of these segments were calculated. The changes of Gibbs free energy reflecting the compatibility of segments and their tendency to the phase separation and the volume fraction of mesogenic rigid core reflecting their ability to arrangement in one direction appear to be crucial in terms of type of the mesophase formation. PMID- 16471859 TI - Substituent effect on the photobleaching of pyrylium salts under ultrashort pulsed illumination. AB - The current article presents the photobleaching properties of a group of pyrylium salts under ultrashort pulsed illumination. These pyrylium salts have the same basic chemical structure and differ only by a specific substituent. It is proven experimentally that two different mechanisms are simultaneously present to the photobleaching of all molecules studied (independently of their specific chemical structure). However, the particular parameters of each mechanism are influenced significantly by the substituent change. The experimental investigation of these parameters showed the presence of multiphoton interactions in the photobleaching of pyrylium salts depending essentially on the specific substituent. PMID- 16471860 TI - Intermolecular energy transfer across nanocrystalline semiconductor surfaces. AB - The yields and dynamics for energy transfer from the metal-to-ligand charge transfer excited states of Ru(deeb)(bpy)(2)(PF(6))(2), Ru(2+), and Os(deeb)(bpy)(2)(PF(6))(2), Os(2+), where deeb is 4,4'-(CH(3)CH(2)CO(2))(2)-2,2' bipyridine, anchored to mesoporous nanocrystalline (anatase) TiO(2) thin films were quantified. Lateral energy transfer from Ru(2+)* to Os(2+) was observed, and the yields were measured as a function of the relative surface coverage and the external solvent environment (CH(3)CN, THF, CCl(4), and hexanes). Excited-state decay of Ru(2+)*/TiO(2) was well described by a parallel first- and second-order kinetic model, whereas Os(2+)*/TiO(2) decayed with first-order kinetics within experimental error. The first-order component was assigned to the radiative and nonradiative decay pathways (tau = 1 micros for Ru(2+)*/TiO(2) and tau = 50 ns for Os(2+)*/TiO(2)). The second-order component was attributed to intermolecular energy transfer followed by triplet-triplet annihilation. An analytical model was derived that allowed determination of the fraction of excited-states that follow the two pathways. The fraction of Ru(2+)*/TiO(2) that decayed through the second order pathway increased with surface coverage and excitation intensity. Monte Carlo simulations were performed to estimate the Ru(2+)* --> Ru(2+) intermolecular energy transfer rate constant of (30 ns)(-1). PMID- 16471861 TI - Influence of Mn-O-Mn bond angle on the magnetic and electronic properties in YBaMn2O5. AB - The influence of the Mn-O-Mn bond angle on the magnetic and electronic properties of YBaMn(2)O(5) was studied by density functional theory, which was implemented in the CASTEP code. In practical calculation, both G- and A-type antiferromagnetic (AFM) orderings were considered. The calculated results indicated that G-type is more stable than A-type, in agreement with both experiment and previous theoretical study. It is also interesting to note that a transition from G-type to A-type at an Mn-O-Mn angle of ca. 170 degrees was found upon increasing Mn-O-Mn angle. Therefore, the calculation suggested that what is essential to stabilize the G-type AFM state is the reduction of the Mn-O-Mn bond angle. For both magnetic orderings, the compound changes from semiconductor to metal with the increase of Mn-O-Mn angle. PMID- 16471862 TI - Regulation of the extent and dynamics of excited-state proton transfer in 2-(2' pyridyl)benzimidazole in nafion membranes by cation exchange. AB - The effect of the microenvironment of a Nafion membrane on the excited-state proton transfer (ESPT) of 2-(2'-pyridyl)benzimidazole (2PBI) has been investigated by steady-state and time-resolved fluorescence spectroscopy. The mechanism of the ESPT is found to depend remarkably on the water content of the membrane. In the protonated form of the membrane, ESPT is found to involve the dicationic (D) form of the fluorophore, whereas in cation-exchanged membranes, it is found to involve the monocation (C). The change in the mechanism and extent of ESPT in cation-exchanged membranes can be explained by considering dehydration of the membrane as well as the less acidic environment around the 2PBI molecules. The slow dynamics is found to result from two factors, namely, slow and incomplete solvation of the transition state, leading to a slowing down of the proton-transfer process, and a slow solvation of the polar tautomeric excited state. PMID- 16471863 TI - Two-dimensional simulations of CuPc-PCTDA solar cells: the importance of mobility and molecular pi stacking. AB - An existing two-dimensional microkinetical model for the photovoltaic effect of molecular-based solar cells has been extended to include electron-hole pair recombination between donor and acceptor sites. Simulations of the short circuit current for simple two-dimensional model heterojunction structures composed of copper phthalocyanine (CuPc) and 3,4,9,10-perylenetetracarboxylic dianhydride (PTCDA) are presented. The short circuit current was investigated as a function of the thickness of the photoactive layer for different choices of mobility for CuPc and PTCDA. The hole mobility of CuPc and/or the electron mobility of PTCDA limits the photovoltaic performance if chosen below a certain threshold determined by the net electron-hole generation rate at the CuPc-PTCDA interface. Also, the mobilities should be of the same order of magnitude. The effect of changing the interplanar separation alpha between the pi stacking molecules was investigated, and it was found that increasing alpha from 0.33 to 0.6 nm increases the short circuit current up to 5 orders of magnitude. This was rationalized in terms of the charge separation energetics of geminate electron hole pairs and its dependence on alpha. As mobilities decrease with increasing alpha and thus opposes this effect, an optimum for alpha approximately 0.66 nm was found for the CuPc-PTCDA heterojunction model structures. The simulations are interpreted in a simple kinetic picture of an electron-hole pair generation step at the CuPc-PTCDA interface and subsequent transport in the CuPc and PTCDA domains. It is argued that an optimal device configuration involves an amorphous region at the CuPc-PTCDA interface and a gradual increase of the molecular order as the electrodes are approached in the respective CuPc and PTCDA transport regions. PMID- 16471864 TI - Study of the semidilute solutions of poly(N,N-dimethylacrylamide) by fluorescence and its implications to the kinetics of coil-to-globule transitions. AB - The large scale motions of poly(N,N-dimethylacrylamide) chains randomly labeled with pyrene (Py-PDMA) were monitored by steady-state and time-resolved fluorescence in semidilute solutions of naked PDMA in acetone and DMF for polymer concentrations ranging from 0 to 550 g/L. Although increasing the polymer concentration of the solution led to a decrease of the mobility of the chromophore attached onto the PDMA backbone, this reduction was rather modest when compared to the large increase of the macroscopic viscosity. This result indicated that locally, the monomer constituting the chains experienced freedom of movement despite the high solution viscosity. The restricted mobility of the chromophore was characterized by the number of monomers occupying the volume probed by the excited chromophore during its lifetime, referred to as a fluorescence "blob". The number of monomers constituting a fluorescence blob, N(F)(-)(blob), and the volume of a fluorescence blob, V(F)(-)(blob), were found to decrease as the polymer concentration of the solution increased, reflecting the decreased mobility experienced by the chromophore. In DMF, the radius of an F blob was found to scale as N(nu)(F)(-)blob, where nu equaled 0.66 +/- 0.03, very close to the expected value of the Flory exponent of 0.6 for a polymer in a good solvent. The combined knowledge of how N(F)(-)(blob) varies with the fluorescence lifetime of the chromophore and the coil density of the polymer was used to propose a new means of studying coil-to-globule transitions with potential implications for predicting the rate of protein folding. PMID- 16471865 TI - Encapsulation mechanism of molecular nanocarriers based on unimolecular micelle forming dendritic core-shell structural polymers. AB - A series of dendritic core-shell structural polymers with different shell densities were synthesized based on dendritic polyester Boltorn H40 and were demonstrated to form unimolecular micelles in chloroform. The encapsulation mechanism study using Congo red as a guest molecule by fluorescence and UV-vis methods showed that the interaction of Congo red with hydroxyl groups in the dendritic core-shell polymer led to encapsulation. Moreover, the results also indicated that the dendrimer with 43.4% hydroxyl groups end-capped by the long alkyl chains showed the best encapsulation capacity. However, lower and higher alkyl densities both led to lower encapsulation capacities. The former was attributed to the poor compatibility of the polymer with chloroform, and the later was caused by less location sites for the guest molecule inside the core shell polymer. PMID- 16471866 TI - Electrochemical polymerization of fluoranthene and characterization of its polymers. AB - A novel inherently conducting polymer, high-quality polyfluoranthene (PFA) film with electrical conductivity of 10(-2) S cm(-1), was first synthesized electrochemically by direct anodic oxidation of fluoranthene in a middle strong Lewis acid-boron trifluoride diethyl etherate. The oxidation potential onset of fluoranthene in this medium was measured to be only 1.07 V vs SCE, which was much lower than that in acetonitrile + 0.1 mol L(-1) tetrabutylammonium tetrafluoroborate (1.68 V vs SCE). This PFA film showed good redox activity and stability even in concentrated sulfuric acid. Moreover, the fluorescence properties of PFA were greatly improved in comparison with those of the monomer. Dedoped PFA films were partly soluble in polar solvents such as CH(2)Cl(2), acetone, tetrahedrofuran, and dimethyl sulfoxide. The structure and morphology of the polymer were investigated by UV-vis spectroscopy, infrared spectroscopy, and scanning electron microscopy, respectively. The results of quantum chemistry calculations of fluoranthene monomer and (1)H NMR spectroscopy of dedoped PFA films indicated that the polymerization mainly occurred at C((3)), C((4)), C((13)), and C((14)) positions. PMID- 16471867 TI - Soluble complexes of sodium poly(isoprene-b-methacrylate) micelles with cationic surfactants in aqueous media. AB - Water-soluble complexes between sodium poly(isoprene-b-methacrylate) (NaIMA) amphiphilic block copolymer micelles and two cationic surfactants with different hydrophobic tail lengths, namely, dodecyltrimethylammonium bromide (DTMAB) and octyltrimethylammonium bromide (OTMAB), were prepared by mixing individual aqueous solutions of block copolymers and surfactants. The complexes were characterized in terms of size, overall charge, and micropolarity by dynamic light scattering, zeta-potential measurements, and fluorescence spectroscopy. Properties of the systems were investigated as a function of surfactant concentration and surfactant type and state in the initial solutions, as well as temperature. Experiments reveal surfactant complexation at the coronal sodium poly(methacrylate) (NaMA) chains, followed by an increase in mass and a decrease in size of the micelles. Complexation of individual surfactant micelles was observed when the DTMAB concentration in the starting solutions was higher than the surfactant cmc. The complexes show a temperature dependence of their dimension due to the hydrophobic effect. PMID- 16471868 TI - Synthesis of large-pore methylene-bridged periodic mesoporous organosilicas and its implications. AB - In this article, we report the synthesis of methylene-bridged periodic mesoporous organosilicas (PMOs) of the SBA-15 type. The materials were characterized by SAXS, BET, NMR, FESEM, and TEM. It was found that the synthesis of methylene bridged SBA-15 PMOs requires more rigorous conditions than that of SBA-15 PMOs bearing organic bridges other than methylene. A mild acidic environment, which slows down the hydrolysis and condensation rates of the precursor, with the assistance of a salt, which enhances precursor-template interaction, should be used to synthesize high-quality large-pore methylene-bridged PMOs. We attributed this to the fast hydrolysis and condensation rates and the rigid backbone of precursor 1,2-bis(triethoxysilyl)methylene. By examining and comparing the synthesis of three large-pore PMOs with different bridges, we concluded that the inductive, bridging, and conformation effects of the organic bridging group play an important role in the synthesis of large-pore PMO materials. PMID- 16471869 TI - Spectroelectrochemistry of poly(ethylenedithiathiophene)--the sulfur analogue of poly(ethylenedioxythiophene). AB - Poly(3,4-ethylenedithiathiophene) (PEDTT) is a polythiophene-like conjugated polymer in which each thiophene ring is functionalized with an ethylenedithia bridge. As such, PEDTT is the sulfur analogue of the well-known poly(3,4 ethylenedioxythiophene) (PEDOT). Substituent effects, namely the presence of sulfur atoms in PEDTT replacing the oxygen atoms of PEDOT, do not provide a simple explanation for the different electronic properties of the two polymers in the neutral state. This paper reports the spectroscopic properties of PEDTT, studied by in situ techniques such as IR-, Vis-, and electron spin resonance (ESR) spectroelectrochemistry. The differences observed upon electrochemical oxidation of PEDTT and PEDOT (e.g., the different infrared active vibrational band patterns in IR spectroelectrochemistry as well as the different nature of the charged states) are even more marked than those observed in the neutral state. These results, with AM1 calculations, indicate conformational effects as a possible explanation for the different electronic and spectroscopic properties of PEDTT and PEDOT. PMID- 16471870 TI - Monoclinic structured BiVO4 nanosheets: hydrothermal preparation, formation mechanism, and coloristic and photocatalytic properties. AB - Bismuth vanadate (BiVO(4)) nanosheets have been hydrothermally synthesized in the presence of sodium dodecyl benzene sulfonate (SDBS) as a morphology-directing template. The nanosheets were characterized by X-ray diffraction (XRD), field emission scanning electron microscopy (FE-SEM) equipped with an X-ray energy dispersive spectrometer (EDS), X-ray photoelectron spectroscopy (XPS), IR spectroscopy, transmission electron microscopy (TEM), and high-resolution TEM (HR TEM). The BiVO(4) nanosheets had a monoclinic structure, were ca. 10-40 nm thick, and showed a preferred (010) surface orientation. The formation mechanism and the effects of reaction temperature and time on the products were investigated. UV visible diffuse reflection spectra indicated that the BiVO(4) nanosheets had outstanding spectral selectivity and improved color properties compared with the corresponding bulk materials. Furthermore, the nanosheets showed good visible photocatalytic activities as determined by degradation of N,N,N',N' tetraethylated rhodamine (RB) under solar irradiation. PMID- 16471871 TI - Effect of pH on the adsorption and activity of creatine phosphokinase. AB - The combination of in situ ellipsometry with atomic force microscopy in the liquid for the study of adsorption of creatine phosphokinase (CPK) onto silicon wafers was shown for the first time. The thickness, adsorbed amount, and topographic information of the adsorbed CPK layers were obtained under different pH conditions. The thickness values of adsorbed CPK layer determined by both techniques were in excellent agreement. At pH 4, CPK monomers present in solution adsorb, forming a very thin (approximately 0.8 nm) layer, indicating CPK unfolding. Upon increasing the pH to 6.8, the adsorbed layer is composed of a mixture of CPK dimers (native structure) and intermediates, increasing the film thickness (approximately 2.4 nm). At pH 9, CPK dimers form monolayers with the highest thickness (approximately 4.0 nm). The nature of interactions between CPK and Si wafers associated with the hydration force seems to control the degree of CPK unfolding upon adsorbing. The enzymatic activity of free CPK and of adsorbed CPK at pH 4, pH 6.8, and pH 9 was measured as a function of pH. In comparison to free CPK in solution, adsorbed CPK presented a shift of the optimal pH from 6.8 toward alkaline pH. PMID- 16471872 TI - Voltammetric and electrochemical ESR studies of oxidation reactions mediated by tris(4-bromophenyl)amine in acetonitrile. AB - The electrochemical oxidation of tris(4-bromophenyl)amine in the presence of 2,6 lutidine is examined in acetonitrile. Voltammetric and spectroscopic investigations suggest that the electrogenerated triaryl aminium radical cation oxidizes 2,6-lutidine in an EC' mechanism, and an equilibrium constant for this homogeneous electron transfer is estimated. The mediated oxidation of a protected phenyl selenoglycoside by this reaction mixture is studied by the use of electrochemical ESR, employing a tubular flow cell, and signal intensity data is found to be consistent with the proposed mechanism, allowing the determination of kinetic parameters by computational simulation. Products of the mediated glycoside oxidation are determined by proton NMR and mass spectrometry. PMID- 16471873 TI - Ionization energy and electronic structure of polycytidine. AB - Ribonucleic acid (RNA) polycytidine (poly rC) homopolymer thin films were prepared on highly oriented pyrolytic graphite (HOPG) substrates. The films were grown from aqueous solution directly in a vacuum in multiple steps with use of an electrospray (ES) deposition system. Before poly rC deposition and after each deposition step the sample was characterized with X-ray and ultraviolet photoemission spectroscopy (XPS, UPS). Evaluation of the UP-spectra sequence allowed the determination of ionization energy and highest occupied molecular orbital (HOMO) electronic structure, as well as the charge injection barriers between HOPG and poly rC. Comparison with earlier results on polyadenosine (poly rA) indicates significant differences between ionization energies (poly rC: 8.1 eV; poly rA: 6.8 eV) and orbital alignment at the graphite interface. The larger ionization energy of poly rC results in a larger hole injection barrier and a smaller electron injection barrier relative to the HOPG Fermi level. PMID- 16471874 TI - Study of the elementary processes involved in the selective oxidation of methane over MoOx/SiO2. AB - Isolated molybdate species supported on silica are reported to have the highest specific activity and selectivity for the direct oxidation of methane to formaldehyde. The present investigation was undertaken to understand the elementary redox processes involved in the formation of formaldehyde over such species. A MoO(x)/SiO(2) catalyst was prepared with a Mo loading of 0.44 Mo/nm(2). On the basis of evidence from extended X-ray absorption fine structure (EXAFS) and Raman spectroscopy, the Mo atoms in this catalyst are present as isolated, pentacoordinated molybdate species containing a single Mo=O bond. Isotopic labeling experiments in combination with in-situ Raman spectroscopy were used to examine the reducibility of the dispersed molybdate species and the exchange of O atoms between the gas phase and the catalyst. It was established that treatment of MoO(x)/SiO(2) at 873 K under pure methane reduces the dispersed molybdate species to only a limited extent and results mainly in the deposition of amorphous carbon. During CH(4) oxidation to formaldehyde, the catalyst undergoes only a very small degree of reduction and typically only approximately 50-500 ppm of Mo(VI) is reduced to Mo(IV). Reactions carried out using CH(4) and (18)O(2) show that there is extensive scrambling of O atoms between the species in the gas phase and the catalyst. Additional experiments revealed that H(2)O formed in the reaction is the principal species responsible for the exchange of O atoms between the gas phase and the SiO(2) support. Low concentrations of H(2)O were observed to enhance the activity of MoO(x)/SiO(2) for CH(4) oxidation to formaldehyde. A mechanism for the oxidation of CH(4) over MoO(x)/SiO(2) was formulated in light of the observations made here and is discussed in the light of previous studies. It is proposed that peroxides are produced by the reaction of O(2) with a small concentration of reduced molybdate species and that the reaction of CH(4) with these peroxide species leads to the formation of formaldehyde. The proposed mechanism also accounts for the positive effects of low concentrations of H(2)O on the rate of formaldehyde formation. PMID- 16471875 TI - Medium effects for very fast electron transfer reactions at electrodes: the [Ru(NH3)6]3+/2+ system in water. AB - The reduction kinetics of [Ru(NH(3))(6)](3+) was studied at Au(111) and Au(100) single-crystal ultramicroelectrodes in dilute perchloric acid electrolytes. Both heterogeneous rate constants and experimental transfer coefficients varied with the crystallographic orientation of the gold surface. The value of the heterogeneous rate constant at Au(111) was significantly larger than that at Au(100). The experimental transfer coefficients also increased but in the opposite order. Standard rate constants at both electrodes increased with an increase in electrolyte concentration. Using double-layer data obtained in 0.01 M HClO(4), it is shown that the true transfer coefficient for this reaction is 0.5 within experimental error. The effective charge on the reactant which has a nominal charge of +3 is close to +1. The latter result reflects the distribution of charge within the polyatomic reactant. PMID- 16471876 TI - Mechanism for interplay between electron and ionic fluxes in KhFek[Fe(CN)6]l.mH2O compounds. AB - This paper develops a framework for the interpretation of ionic insertion/deinsertion reactions in an aqueous environment taking place in transition-metal hexacyanoferrates of the general formula K(h)[Fe(2+) (CN)(6)](l).mH(2)O, also called Prussian Blue. Three different processes were fully separated in the electrochemistry of these films. It was clearly identified that one of these electrochemical processes involves the insertion/deinsertion of H(3)O(+) (hydrated protons) through the channels of the K(h)[Fe(2+) (CN)(6)](l).mH(2)O structure to reach the film electroneutrality during the electron transfer between Everitt's Salt and Prussian Blue. The other electrochemical processes involve K(+) or H(+) (proton) exchange through the water crystalline structure existing in the channels of the K(h)[Fe(2+)(CN)(6)](l).mH(2)O structure. PMID- 16471877 TI - Electron-stimulated production of molecular oxygen in amorphous solid water. AB - The low-energy, electron-stimulated production of molecular oxygen from pure amorphous solid water (ASW) films and ASW films codosed with H(2)O(2) is investigated. Layered films of H(2)(16)O and H(2)(18)O are used to investigate the reaction mechanisms for O(2) production and the spatial profile of the reactions within the films. The O(2) yield is dose-dependent, indicating that precursors are involved in the O(2) production. For temperatures below approximately 80 K, the O(2) yield at steady state is relatively low and nearly independent of temperature. At higher temperatures, the yield increases rapidly. The O(2) yield is enhanced from H(2)O(2)-dosed water films, but the experiments show that H(2)O(2) is not the final precursor in the reactions leading to O(2). Instead, a stable precursor for O(2) is produced through a multistep reaction sequence probably involving the reactions of OH radicals to produce H(2)O(2) and then HO(2). The O(2) is produced in a nonthermal reaction from the HO(2). For relatively thick films, the reactions leading to O(2) occur at or near the ASW/vacuum interface. However, the electronic excitations that initiate the reactions occur over a larger range in the film. A kinetic model that qualitatively accounts for all of the observations is presented. PMID- 16471878 TI - Nature, density, and catalytic role of exposed species on dispersed VOx/CrOx/Al2O3 catalysts. AB - The structure and surface composition of binary oxides consisting of CrO(x) and VO(x) dispersed on alumina and their effects on the rate and selectivity of oxidative dehydrogenation (ODH) of propane were examined and compared with those for CrO(x) and VO(x) dispersed on alumina. VO(x) deposition on an equivalent CrO(x) monolayer on alumina and deposition of CrO(x) on an equivalent monolayer of VO(x) deposited on alumina led to CrVO(4) species during thermal treatment with concomitant reduction of Cr(6+) to Cr(3+). Autoreduction of Cr(6+) to Cr(3+) is also detected for CrO(x), even without the presence of VO(x). Infrared spectroscopy of NO adsorbed at 153 K probes the relative abundance of alumina and of V(5+), Cr(3+), and Cr(6+) at surfaces. This technique detects differences in the surface composition of VO(x)/CrO(x)()/Al(2)O(3) and CrO(x)/VO(x)/Al(2)O(3). The first of these samples is enriched in VO(x) relative to CrO(x) compared with the second sample. Consistent with this finding, VO(x)/CrO(x)/Al(2)O(3) and CrO(x)/VO(x)/Al(2)O(3) are distinguishable in their ODH activities and propene selectivities. The highest ODH activity and propene selectivity is observed for VO(x)/CrO(x)/Al(2)O(3), which exhibits a surface enriched in VO(x) and having a low surface concentration of Cr(6+). PMID- 16471879 TI - TiCl3-enhanced NaAlH4: impact of excess al and development of the Al1-yTiy phase during cycling. AB - NaAlH(4) with TiCl(3) and Al were mixed by ball-milling and cycled three times. The hydrogen storage properties were monitored during cycling, and the products were characterized by synchrotron X-ray diffraction. Because of the previously described formation of Al(1)(-)(y)Ti(y) with y approximately 0.15 during cycling that traps Al beyond the amount associated with the formation of NaCl, some Na(3)AlH(6) has no free Al to react with to form NaAlH(4). This was counteracted in the present work by adding a stoichiometric amount of Al that increases the theoretical storage capacity. Due to limitations in metal diffusion small amounts of Na(3)AlH(6) were still detected. When approximately 7 mol % more Al than the stoichiometric amount was added, the observed storage capacity increased significantly, and the Na(3)AlH(6) content was negligible after prolonged rehydrogenation. Cycled NaAlH(4) + 10 mol % TiCl(3) were desorbed to two different levels, and the diffraction patterns were compared. There is no change in unit-cell dimensions during desorption, and there is no sign of changes in the bulk composition of the Al(1)(-)(y)Ti(y) phase during a cycle. Adding pure Ti to a NaH + Al mixture by ball-milling in argon or hydrogen results in formation of TiH(2) that is stable during at least one cycle. PMID- 16471880 TI - Water density in the electric double layer at the insulator/electrolyte solution interface. AB - I studied the spatial structure of the thick transition region between n-hexane and a colloidal solution of 7-nm silica particles by X-ray reflectivity and grazing incidence small-angle scattering. The interfacial structure is discussed in terms of a semiquantitative interface model wherein the potential gradient at the n-hexane/sol interface reflects the difference in the potentials of "image forces" between the cationic Na(+) and anions (nanoparticles) and the specific adsorption of surface charge at the interface between the adsorbed layer and the solution, as well as at the interface between the adsorbed layer and n-hexane. The X-ray scattering data revealed that the average density of water in the field approximately 10(9)-10(10) V/m of the electrical double layer at the hexane/silica sol interface is the same as, or only few percent higher (1-7%) than, its density under normal conditions. PMID- 16471881 TI - Interfacial energy and the law of corresponding states 2: associated fluids. AB - A mesoscopic model for the liquid/vapor interface previously developed for nonpolar fluids [J. Phys. Chem. A 2003, 107, 875; 2003, 107, 883] is extended to the case of polar associated compounds. The interfacial energy is factorized in two terms: one corresponding to association depending on the hydrogen bonds density, the other corresponding to the nonpolar contribution. This last term is treated in the framework of the corresponding states formalism similar to the one used in the case of nonpolar fluids [J. Phys. Chem. B 2004, 108, 5951]. The model yields a generalized behavior of the association factor as a function of the dielectric constant for the treated fluids. The calculated surface tension shows a mean error of about 1% for seven compounds having different multivalent H-bond characters. PMID- 16471882 TI - Dielectric characterization of a nanofiltration membrane in electrolyte solutions: its double-layer structure and ion permeation. AB - Dielectric spectroscopy (DS) was applied to a nanofiltration (NF) membrane to detect its double-layer structure and ion permeation. Dielectric measurements were carried out on the systems composed of the NF membrane NTR7450 and dilute solutions of eight electrolytes, LiCl, NaCl, KCl, NH(4)Cl, MgCl(2), CaCl(2), BaCl(2), and CuCl(2). Two relaxations were observed in the frequency range from 40 Hz to 4 MHz for each system. On the basis of characteristics of the dielectric spectra and the Maxwell-Wagner interfacial polarization theory, the low-frequency relaxation was attributed to inhomogeneity of the membrane structure itself, whereas the high-frequency relaxation was attributed to interfacial polarization between the membrane and the solution. A multiphase dielectric model previously developed by one of the authors and co-workers was adopted to present systems to analyze the dielectric spectra, and electric parameters, i.e., capacitance and conductance, of the two layers composing the membrane were obtained. The electric properties estimated for the two layers were different and changed with the environment in a different manner. Further analyses suggest that the two layers had a different separation mechanism due to their difference in materials, looseness, and fixed charge content. The fixed charge density of one layer was estimated, and the ion permeation difficulties in both layers was compared. This research revealed that DS was by far an effective method to obtain detailed electric parameters about the inner multilayer structure of the NF membrane and to elucidate separation mechanisms of each layer. PMID- 16471883 TI - Two modes of transformation of amorphous calcium carbonate films in air. AB - Large-area amorphous calcium carbonate (ACC) films in air are shown to be transformed into crystalline calcium carbonate (CaCO(3)) films via two modes dissolution-recrystallization and solid-solid phase transition-depending on the relative humidity of the air and the temperature. Moisture in the air promotes the transformation of ACC into crystalline forms via a dissolution recrystallization process. Increasing the humidity increases the rate of ACC crystallization and gives rise to films with numerous large pores. As the temperature is increased, the effect of moisture in the air is reduced and solid solid transition by thermal activation becomes the dominant transformation mechanism. At 100 and 120 degrees C, ACC films are transformed into predominantly (110) oriented crystalline films. Collectively, the results show that calcium carbonate films with different morphologies, crystal phases, and structures can be obtained by controlling the humidity and temperature. This ability to control the transformation of ACC should assist in clarifying the role of ACC in the biomineralization of CaCO(3) and should open new avenues for preparing CaCO(3) films with oriented and fine structure. PMID- 16471884 TI - Direct gas-liquid interfacial dynamics: the reaction between O(3P) and a liquid hydrocarbon. AB - We report the first measurements of internal energy distributions of the OH produced via a direct mechanism, isolated from other components on the basis of time-of-flight, in the interfacial reaction between gas-phase O((3)P) atoms and the liquid hydrocarbon squalane, C(30)H(62). O((3)P) atoms were generated by laser photolysis of NO(2) above the liquid. Resulting hydroxyl radicals that escape from the surface were detected by laser-induced fluorescence. Time-of flight profiles demonstrate that the kinetic energy of the fastest OH (nu' = 1) is lower than that of (nu' = 0). Rotational distributions were measured at the rising edge of their appearance for both OH (nu' = 0) and (nu' = 1). They were found to differ substantially more than at the peak of their profiles. They were also less dependent on the bulk liquid temperature. We conclude that the new data confirm strongly that at least two mechanisms contribute to the production of OH. The higher-velocity component has translational and rotational energy distributions, observed cleanly for the first time, consistent with a direct mechanism. The close correspondence of these rotational distributions to those from the corresponding homogeneous gas-phase reaction of O((3)P) with smaller hydrocarbons suggests a very similar, near collinear direct abstraction. This is accompanied by a slower component with kinetic energy and rotational (but not vibrational) distributions reflecting the temperature of the liquid, consistent with a distinct trapping-desorption mechanism. PMID- 16471885 TI - Effect of water on the molecular structure and arrangement of nitrile functionalized ionic liquids. AB - Water changes the physical properties of ionic liquids (ILs). In this paper, the effect of water on ILs is investigated at the molecular level. The molecular structure and arrangement of 1-butyronitrile-3-methylimidazolium halide, in the presence and absence of the intruded water molecule, have been elucidated by single-crystal X-ray crystallography and near-infrared Raman spectroscopy. Water molecule is found to change the conformation of the n-butyronitrile chain of the cation. The hydrogen bonding interaction between the anion and water molecule, leading to loose molecular packing, is most likely to be responsible for the change. Distinct molecular structures and arrangements of ILs with and without water molecule have been presented here for the first time. As the unique properties of ILs are related to their structures and molecular arrangements, the presence of water, wanted or unwanted, must be carefully examined in any kind of IL research and applications. PMID- 16471887 TI - Surface structure and interface dynamics of alkanethiol self-assembled monolayers on Au(111). AB - Scanning tunneling microscopy (STM) and high-resolution electron energy loss spectroscopy (HREELS) were used to examine the structural transitions and interface dynamics of octanethiol (OT) self-assembled monolayers (SAMs) caused by long-term storage or annealing at an elevated temperature. We found that the structural transitions of OT SAMs from the c(4 x 2) superlattice to the (6 x square root 3) superlattice resulting from long-term storage were caused by both the dynamic movement of the adsorbed sulfur atoms on several adsorption sites of the Au(111) surface and the change of molecular orientation in the ordered layer. Moreover, it was found that the chemical structure of the sulfur headgroups does not change from monomer to dimer by the temporal change of SAMs at room temperature. Contrary to the results of the long-term-stored SAMs, it was found that the annealing process did not modify either the interfacial or chemical structures of the sulfur headgroups or the two-dimensional c(4 x 2) domain structure. Our results will be very useful for a better understanding of the interface dynamics and stability of sulfur atoms in alkanethiol SAMs on Au(111) surfaces. PMID- 16471886 TI - Development of an empirical force field for silica. Application to the quartz water interface. AB - Interactions of pulverized crystalline silica with biological systems, including the lungs, cause cell damage, inflammation, and apoptosis. To allow computational atomistic modeling of these pathogenic processes, including interactions between silica surfaces and biological molecules, new parameters for quartz, compatible with the CHARMM empirical force field were developed. Parameters were optimized to reproduce the experimental geometry of alpha-quartz, ab initio vibrational spectra, and interactions between model compounds and water. The newly developed force field was used to study interactions of water with two singular surfaces of alpha-quartz, (011) and (100). Properties monitored and analyzed include the variation of the density of water molecules in the plane perpendicular to the surface, disruption of the water H-bond network upon adsorption, and space-time correlations of water oxygen atoms in terms of Van Hove self-correlation functions. The vibrational density of states spectra of water in confined compartments were also computed and compared with experimental neutron-scattering results. Both the attenuation and shifting to higher frequencies of the hindered translational peaks upon confinement are clearly reproduced by the model. However, an upshift of librational peaks under the conditions of model confinement still remains underrepresented at the current empirical level. PMID- 16471888 TI - Fabrication and electrochemical application of three-dimensional gold nanoparticles: self-assembly. AB - Multilayers film of nanostructured citrate-stabilized gold particles (AuNPs) has been fabricated based on the layer-by-layer (LBL) technique using a self assembled monolayer of 1,4-benzenedimethanethiol (BDMT). The formation of AuNPs and BDMT self-assemblies as alternative multilayers was confirmed by transmission electron microscopy (TEM), X-ray photoelectron spectroscope (XPS), and quartz crystal microbalance (QCM). The formation of uniform AuNP layers with an average monolayer thickness of 5-6 nm was obvious in the TEM images. The existence of BDMT molecules as cross linkers for the AuNPs' layers was proved by XPS measurements. The greater affinity of AuNPs' layers to bind BDMT molecules in comparison with the bare Au bulk electrode was revealed by QCM measurements. Electrochemically, the AuNPs' layers on the electrode surface did not only catalyze the reduction of oxygen (ca. 100-mV positive shift of the reduction peak potential compared with that at the bare Au bulk electrode) but also showed a fascinating nature of working as a renewed activated-electrode surface; a zigzag response was observed for oxygen reduction during alternative immobilization of BDMT and the AuNP layer. The self-assembly of a new AuNPs layer restored the catalytic activity that was entirely blocked by the preceding BDMT layer. PMID- 16471889 TI - Density functional theory study of formic acid adsorption on anatase TiO2(001): geometries, energetics, and effects of coverage, hydration, and reconstruction. AB - We present density functional theory calculations and first-principles molecular dynamics simulations of formic acid adsorption on anatase TiO(2)(001), the minority surface exposed by anatase TiO(2) nanoparticles. A wide range of factors that may affect formic acid adsorption, such as coverage, surface hydration, and reconstruction, are considered. It is found that (i) formic acid dissociates spontaneously on unreconstructed clean TiO(2)(001)-1 x 1, as well as on the highly reactive ridge of the reconstructed TiO(2)(001)-1 x 4 surface; (ii) on both the 1 x 1 and 1 x 4 surfaces, various configurations of dissociated formic acid exist with adsorption energies of about 1.5 eV, which very weakly depend on the coverage; (iii) bidentate adsorption configurations, in which the formate moiety binds to the surface through two Ti-O bonds, are energetically more favored than monodentate ones; (iv) partial hydration of TiO(2)(001)-1 x 1 tends to favor the bidentate chelating configuration with respect to the bridging one but has otherwise little effect on the adsorption energetics; and (v) physical adsorption of formic acid on fully hydrated TiO(2)(001)-1 x 1 is also fairly strong. Comparison of the present results for formic acid adsorption with those for water and methanol under similar conditions provides valuable insights to the understanding of recent experimental results concerning the coadsorption of these molecules. PMID- 16471890 TI - Simple electrochemical method for deposition and voltammetric inspection of silver particles at the liquid-liquid interface of a thin-film electrode. AB - A novel experimental methodology for depositing and voltammetric study of Ag nanoparticles at the water-nitrobenzene (W-NB) interface is proposed by means of thin-film electrodes. The electrode assembly consists of a graphite electrode modified with a thin NB film containing decamethylferrocene (DMFC) as a redox probe. In contact with an aqueous electrolyte containing Ag(+) ions, a heterogeneous electron-transfer reaction between DMFC((NB)) and Ag(+)((W)) takes place to form DMFC(+)((NB)) and Ag deposit at the W-NB interface. Based on this interfacial reaction, two different deposition strategies have been applied. In the uncontrolled potential deposition protocol, the electrode is immersed into an AgNO(3) aqueous solution for a certain period under open circuit conditions. Following the deposition step, the Ag-modified thin-film electrode is transferred into an aqueous electrolyte free of Ag(+) ions and voltammetrically inspected. In the second protocol the deposition was carried out under controlled potential conditions, i.e., in an aqueous electrolyte solution containing Ag(+) ions by permanent cycling of the electrode potential. In this procedure, DMFC((NB)) is electrochemically regenerated at the electrode surface, hence enabling continuation and voltammetric control of the Ag deposition. Hence, the overall electrochemical process can be regarded as an electrochemical reduction of Ag(+)((W)) at the W-NB interface, where the redox couple DMFC(+)/DMFC acts as a mediator for shuttling electrons from the electrode to the W-NB interface. Ag particles deposited at the W-NB interface affect the ion transfer across the interface, which provides the basis for voltammetric inspection of the metal deposit at the liquid-liquid interface with thin-film electrodes. Voltammetric properties of thin-film electrodes are particularly sensitive to the deposition procedure, reflecting differences in the properties of the Ag deposit. Moreover, this methodology is particularly suited to inspect catalytic activities of metal particles deposited at the liquid-liquid interface toward heterogeneous electron transfer reactions occurring at the at the liquid-liquid interface. PMID- 16471891 TI - Molecular modeling and experimental studies of the thermodynamic and transport properties of pyridinium-based ionic liquids. AB - A combined experimental and molecular dynamics study has been performed on the following pyridinium-based ionic liquids: 1-n-hexyl-3-methylpyridinium bis(trifluoromethanesulfonyl)imide ([hmpy][Tf(2)N]), 1-n-octyl-3-methylpyridinium bis(trifluoromethanesulfonyl)imide ([ompy][Tf(2)N]), and 1-n-hexyl-3,5 dimethylpyridinium bis(trifluoromethanesulfonyl)imide ([hdmpy][Tf(2)N]). Pulsed field gradient nuclear magnetic resonance spectroscopy was used to determine the self-diffusivities of the individual cations and anions as a function of temperature. Experimental self-diffusivities range from 10(-11) to 10(-10) m(2)/s. Activation energies for diffusion are 44-49 kJ/mol. A classical force field was developed for these compounds, and molecular dynamics simulations were performed to compute dynamic as well as thermodynamic properties. Evidence of glassy dynamics was found, preventing accurate determination of self diffusivities over molecular dynamics time scales. Volumetric properties such as density, isothermal compressibility, and volumetric expansivity agree well with experiment. Simulated heat capacities are within 2% of experimental values. PMID- 16471892 TI - Physicochemical properties and structures of room-temperature ionic liquids. 3. Variation of cationic structures. AB - A series of room-temperature ionic liquids (RTILs) were prepared with different cationic structures, 1-butyl-3-methylimidazolium ([bmim]), 1-butylpyridinium ([bpy]), N-butyl-N-methylpyrrolidinium, ([bmpro]), and N-butyl-N,N,N trimethylammonium ([(n-C(4)H(9))(CH(3))(3)N]) combined with an anion, bis(trifluoromethane sulfonyl)imide ([(CF(3)SO(2))(2)N]), and the thermal property, density, self-diffusion coefficients of the cation and anion, viscosity, and ionic conductivity were measured over a wide temperature range. The self-diffusion coefficient, viscosity, ionic conductivity, and molar conductivity follow the Vogel-Fulcher-Tamman equation for temperature dependencies, and the best-fit parameters have been estimated, together with the linear fitting parameters for the density. The relative cationic and anionic self diffusion coefficients for the RTILs, independently determined by the pulsed field-gradient spin-echo NMR method, appear to be influenced by the shape of the cationic structure. A definite order of the summation of the cationic and anionic diffusion coefficients for the RTILs: [bmim][(CF(3)SO(2))(2)N] > [bpy][(CF(3)SO(2))(2)N] > [bmpro][(CF(3)SO(2))(2)N] > [(n C(4)H(9))(CH(3))(3)N][(CF(3)SO(2))(2)N], has been observed, which coincides with the reverse order to the viscosity data. The ratio of molar conductivity obtained from the impedance measurements to that calculated by the ionic diffusivity using the Nernst-Einstein equation quantifies the active ions contributing to ionic conduction in the diffusion components and follows the order: [bmpro][(CF(3)SO(2))(2)N] > [(n-C(4)H(9))(CH(3))(3)N][(CF(3)SO(2))(2)N] > [bpy][(CF(3)SO(2))(2)N] > [bmim][(CF(3)SO(2))(2)N] at 30 degrees C. PMID- 16471893 TI - Gas hydrates of argon and methane synthesized at high pressures: composition, thermal expansion, and self-preservation. AB - For the first time, the compositions of argon and methane high-pressure gas hydrates have been directly determined. The studied samples of the gas hydrates were prepared under high-pressure conditions and quenched at 77 K. The composition of the argon hydrate (structure H, stable at 460-770 MPa) was found to be Ar.(3.27 +/- 0.17)H(2)O. This result shows a good agreement with the refinement of the argon hydrate structure using neutron powder diffraction data and helps to rationalize the evolution of hydrate structures in the Ar-H(2)O system at high pressures. The quenched argon hydrate was found to dissociate in two steps. The first step (170-190 K) corresponds to a partial dissociation of the hydrate and the self-preservation of a residual part of the hydrate with an ice cover. Presumably, significant amounts of ice Ic form at this stage. The second step (210-230 K) corresponds to the dissociation of the residual part of the hydrate. The composition of the methane hydrate (cubic structure I, stable up to 620 MPa) was found to be CH(4).5.76H(2)O. Temperature dependence of the unit cell parameters for both hydrates has been also studied. Calculated from these results, the thermal expansivities for the structure H argon hydrate are alpha(a) = 76.6 K(-1) and alpha(c) = 77.4 K(-1) (in the 100-250 K temperature range) and for the cubic structure I methane hydrate are alpha(a) = 32.2 K(-1), alpha(a) = 53.0 K(-1), and alpha(a) = 73.5 K(-1) at 100, 150, and 200 K, respectively. PMID- 16471894 TI - Modeling a liquid crystal dynamics by atomistic simulation with an ab initio derived force field. AB - Atomistic molecular dynamics (MD) simulations of 4-n-pentyl 4'-cyano-biphenyl (5CB) have been performed, adopting a specific ab initio derived force field. Two state points in the nematic phase and three in the isotropic phase, as determined in a previous work, have been considered. At each state point, at least 10 ns have been produced, allowing us to accurately calculate single-molecule properties. In the isotropic phase, the values of the translational diffusion coefficient, and even more so the activation energy for the process, agree well with experimental data. Qualitatively, also the dynamic anisotropy of the nematic phase is correctly accounted for. Rotational diffusion coefficients, which describe spinning and tumbling motions, fall well within the range of experimental values. The reorientational dynamics of our model 5CB covers diverse time regimes. The longest one is strongly temperature dependent and characterized by a relaxation time in accord with experimental dielectric relaxation data. Shear viscosity and Landau-de Gennes relaxation times, typically collective variables, reproduce the experimental results very well in the isotropic phase. In the nematic phase, despite a large statistical uncertainty due to the extremely slow relaxation of the correlation functions involved, our simulation yields the correct relative order of the three experimental Miesowicz viscosities. PMID- 16471895 TI - Electric polarizability of DNA in aqueous salt solution. AB - Monte Carlo simulations are performed to determine the anisotropy of the electric polarizability of a model DNA fragment in aqueous salt solution. By taking into consideration the participation of coions in the electroneutrality condition, at every simulation step, we obtain a list of counterions constituting the net charge arranged in increasing order of their distance from the DNA and calculate the contribution to the dipole moment from the first n counterions in the list. We define a partial polarizability tensor due to these n counterions to understand the origin of the polarizability in close relation to the solution structure. The ionic distributions are described by the counterion condensation theory. Characteristic features of the electric properties of polyelectrolytes are reproduced. The anisotropy of the electric polarizability Deltaalpha of DNA decreases with the addition of salt, yielding values comparable to experiment. The effect of electrophoretic motion of the polyion is examined by estimating its upper limit. PMID- 16471896 TI - Equilibrium dialysis data and the relationships between preferential interaction parameters for biological systems in terms of Kirkwood-Buff integrals. AB - Equilibrium dialysis data has provided valuable information concerning the preferential interaction of a cosolvent with a biomolecule in aqueous solutions. Here, we formulate the experimental data in terms of Kirkwood-Buff (KB) theory, resulting in equations that provide a simple physical picture of the dialysis experiment and thereby the interaction of a cosolvent with a biomolecule. These results are then used to establish exact relationships between preferential interaction coefficients, defined in different ensembles and/or using different concentration scales, in terms of KB integrals. It is then argued that the molality based equilibrium dialysis data represent the situation most relevant to computer simulations performed in either open or closed systems. PMID- 16471898 TI - Intermolecular interaction of actin revealed by a dynamic light scattering technique. AB - The intermolecular interaction force of actin was studied by a dynamic light scattering technique. The mutual diffusion coefficients (D) of monomeric actin were accurately determined in a G-buffer with a low concentration of KCl from 0 to 10 mM. The translational diffusion coefficient was obtained as D(0) = (87 +/- 3) x 10(-12) m(2).s(-1) at 25 degrees C and pH 7.4, which gives a hydrodynamic radius of monomeric actin of r(H) = 2.8 +/- 0.1 nm. The Derjaguin-Landau-Verwey Overbeek (DLVO) theory, assuming electrostatic and van der Waals potentials, failed to describe the change in interaction parameter (lambda) with KCl concentration, but the extended DLVO theory succeeded if an additional repulsive potential was assumed. The Hamaker constant of actin in the Ca(2+)-ATP bound state was determined for the first time as A(H) = 10.4 +/- 0.6 k(B)T. PMID- 16471897 TI - Optimization of the GB/SA solvation model for predicting the structure of surface loops in proteins. AB - Implicit solvation models are commonly optimized with respect to experimental data or Poisson-Boltzmann (PB) results obtained for small molecules, where the force field is sometimes not considered. In previous studies, we have developed an optimization procedure for cyclic peptides and surface loops in proteins based on the entire system studied and the specific force field used. Thus, the loop has been modeled by the simplified solvation function E(tot) = E(FF) (epsilon = 2r) + Sigma(i) sigma(i)A(i), where E(FF) (epsilon = nr) is the AMBER force field energy with a distance-dependent dielectric function, epsilon = nr, A(i) is the solvent accessible surface area of atom i, and sigma(i) is its atomic solvation parameter. During the optimization process, the loop is free to move while the protein template is held fixed in its X-ray structure. To improve on the results of this model, in the present work we apply our optimization procedure to the physically more rigorous solvation model, the generalized Born with surface area (GB/SA) (together with the all-atom AMBER force field) as suggested by Still and co-workers (J. Phys. Chem. A 1997, 101, 3005). The six parameters of the GB/SA model, namely, P(1)-P(5) and the surface area parameter, sigma (programmed in the TINKER package) are reoptimized for a "training" group of nine loops, and a best fit set is defined from the individual sets of optimized parameters. The best-fit set and Still's original set of parameters (where Lys, Arg, His, Glu, and Asp are charged or neutralized) were applied to the training group as well as to a "test" group of seven loops, and the energy gaps and the corresponding RMSD values were calculated. These GB/SA results based on the three sets of parameters have been found to be comparable; surprisingly, however, they are somewhat inferior (e.g, of larger energy gaps) to those obtained previously from the simplified model described above. We discuss recent results for loops obtained by other solvation models and potential directions for future studies. PMID- 16471899 TI - Visualization and characterization of the infrared active amide I vibrations of proteins. AB - To facilitate the analysis of frequency-structure correlations in the amide I vibrational spectroscopy of proteins, we investigate visualization methods and spatial correlation functions that describe delocalized vibrations of proteins and protein secondary structures. To study those vibrational modes revealed in infrared spectroscopy, we characterize frequency-dependent bright states obtained from doorway mode analysis. Our visualization methods pictorially color code amplitude and phase of each oscillator within the structure to reveal spatially varying patterns characteristic of excitations within sheets and helices. Spatial correlation functions in the amplitude and phase of amide I oscillators quantitatively address the extent of delocalization and the alpha helical and beta sheet character of these modes. Specifically, we investigate the vibrations of idealized antiparallel beta sheets and alpha helices and perform case studies on three proteins: concanavalin A, myoglobin, and ubiquitin. PMID- 16471900 TI - Structural fluctuations and excitation transfer between adenine and 2-aminopurine in single-stranded deoxytrinucleotides. AB - Steady-state fluorescence measurements on the deoxytrinucleotides (5')dTp2APpA(3') and (5')dAp2APpA(3') show a temperature-dependence and a viscosity-dependence for energy transfer that qualitatively differ from those seen in our previous study of charge transfer (CT) in these systems. Time resolved anisotropy studies and molecular dynamics simulations are presented that provide a detailed characterization of the structural dynamics of these systems and how these fluctuations modulate the electronic interaction between 2AP and its neighbors. To gain quantitative insight into the interplay of conformational fluctuations and stacking-induced energy transfer, we present results from a new hybrid quantum-classical simulation method for computing the A --> 2AP energy transfer rate that makes use of the full three-dimensional nature of the donor and acceptor transition densities. Analysis of the results shows that the standard transition dipole-transition dipole approximation for the Coulombic coupling substantially overestimates the transfer rate and that the nearest neighbor energy transfer from adenine to 2AP occurs on a much faster time scale than that for CT. This suggests that, unlike the CT dynamics where conformational "gating" plays a critical role, the large amplitude fluctuations that modulate the process are largely "frozen" out on the energy transfer time scale. PMID- 16471901 TI - How can (-)-epigallocatechin gallate from green tea prevent HIV-1 infection? Mechanistic insights from computational modeling and the implication for rational design of anti-HIV-1 entry inhibitors. AB - Possible inhibitors preventing human immunodeficiency virus type 1 (HIV-1) entry into the cells are recognized as hopeful next-generation anti-HIV-1 drugs. It is highly desirable to develop a potent inhibitor blocking binding of glycoprotein CD4 of the cell with glycoprotein gp120 of HIV-1, because the gp120-CD4 binding is the initial step of HIV-1 entry into the cells. It has been recently reported that (-)-epigallocatechin gallate (EGCG) from green tea is an inhibitor blocking gp120-CD4 binding. But the inhibitory mechanism remains unknown. For understanding the inhibitory mechanism, extensive molecular docking, molecular dynamics simulations, and binding free-energy calculations have been performed in this study to predict the most favorable structures of CD4-EGCG, gp120-CD4, and gp120-CD4-EGCG binding complexes in water. The results reveal that EGCG binds with CD4 in such a way that the calculated binding affinity of gp120 with the CD4 EGCG complex is negligible. So, the favorable binding of EGCG with CD4 can effectively block gp120-CD4 binding. The calculated CD4-EGCG binding affinity (DeltaG(bind) = -5.5 kcal/mol, K(d) = 94 microM) is in excellent agreement with available experimental data suggesting IC(50) approximately 100 microM for EGCG blocking CD4-gp120 binding. These results and insights provide a rational basis for future design of novel, more potent inhibitors to block gp120-CD4 binding. PMID- 16471902 TI - Molecular dynamics studies of ion distributions for DNA duplexes and DNA clusters: salt effects and connection to DNA melting. AB - We present extensive molecular dynamics simulations of the ion distributions for DNA duplexes and DNA clusters using the Amber force field with implicit water. The distribution of ions and the electrostatic energy of ions around an isolated DNA duplex and clusters of DNA duplexes in different salt (NaCl) concentrations over the range 0.2-1.0 mol/L are determined on the basis of the simulation results. Using the electrostatic energy profile, we determine a local net charge fraction phi, which is found to increase with increasing of salt concentration. For DNA clusters containing two DNA duplexes (DNA pair) or four DNA duplexes, phi increases as the distance between the duplexes decreases. Combining this result with experimental results for the dependence of the DNA melting temperature on bulk salt concentration, we conclude that for a pair of DNA duplexes the melting temperature increases by 5-10 K for interaxis separations of 25-40 A. For a cluster of four DNA duplexes, an even larger melting temperature increase should occur. We argue that this melting temperature increase in dense DNA clusters is responsible for the cooperative melting mechanism in DNA-linked nanoparticle aggregates and DNA-linked polymer aggregates. PMID- 16471903 TI - Constant-pH molecular dynamics with ionic strength effects: protonation conformation coupling in decalysine. AB - A new implementation of the stochastic titration method for constant-pH molecular dynamics is presented, which introduces ionic strength effects in the simulations. In addition, the new implementation uses a faster molecular dynamics algorithm and an improved treatment of protonation events and of their effect on force field parameters. This new methodology is applied to a decalysine peptide, yielding very good quantitative agreement with experiments, both in terms of titration and helix-coil transition. The results show a significant dependence on ionic strength, illustrating the importance of including this parameter in constant-pH molecular dynamics simulations. Overall, the method seems to properly capture the protonation-conformation coupling and its dependence on ionic strength. PMID- 16471904 TI - Towards accurate ab initio QM/MM calculations of free-energy profiles of enzymatic reactions. AB - Reliable studies of enzymatic reactions by combined quantum mechanical/molecular mechanics (QM/MM) approaches, with an ab initio description of the quantum region, presents a major challenge to computational chemists. The main problem is the need for a large amount of computer time to evaluate the QM energy, which in turn makes it extremely challenging to perform proper configurational sampling. This work presents major progress toward the evaluation of ab initio QM/MM free energy surfaces and activation free energies of reactions in enzymes and in solutions. This is done by exploiting our previous idea of using the empirical valence bond (EVB) method as a reference potential and then using the linear response approximation (LRA) approach to evaluate the free energies of transfer from the EVB to the QM/MM surfaces in the reactant and product state. However, the new crucial step involves the use of a constraint at the transition state that fixes the system at a given value of the reaction coordinate and allows us to use the LRA at the transition state. The advance offered by the present approach is particularly significant because it evaluates the free energy associated with both the substrate and the solvent motions. This evaluation appeared to be a relatively simple task once one uses a classical reference potential. The main problem has been using the reference potential for the evaluation of the free-energy contributions associated with the solute motions where the difference between the reference EVB potential and the QM/MM potential can be large. The present refinement finally allows us to overcome the problems with the solute fluctuations and therefore to obtain, for the first time, a free energy barrier that reflects the solute entropy properly. Thus, we present a way to evaluate the complete QM/MM activation free energy with an equal footing treatment of the solute and the solvent. This provides a general consistent and effective strategy for evaluating the QM/MM activation free energies in proteins and in solution. Our advance allows one to explore consistently various mechanistic and catalytic proposals while using ab initio (ai) QM/MM approaches. PMID- 16471905 TI - Visible photooxidation of dibenzothiophenes sensitized by 2-(4-methoxyphenyl)-4, 6-diphenylpyrylium: an electron transfer mechanism without involvement of superoxide. AB - We report here on a new electron-transfer mechanism for visible-light photooxidation of sulfides in which no superoxide ion is involved. Visible-light irradiation of 2-(4-methoxyphenyl)-4, 6-diphenylpyrylium tetrafluoroborate (MOPDPP(+)BF(4)(-)) in an O(2)-saturated acetonitrile solution containing dibenzothiophene (DBT) results in nearly 100% conversion to oxygenated products, DBT sulfoxide and sulfone. The photooxidation of DBT is initiated by a photoinduced electron-transfer process, where the excited MOPDPP(+) traps an electron from the ground-state DBT to form MOPDPP(*) and DBT radical cation. Such a mechanism is consistent with the studies of laser flash photolysis, electron spin resonance, and fluorescence quenching of the irradiated system. The photogenerated DBT radical cation undergoes a coupling reaction with O(2) to produce the intermediate responsible for the formation of the oxygenated products. The presence of O(2) has no effect on the decay kinetics of the transient absorption of MOPDPP(*), indicating that no redox reaction occurs between MOPDPP(*) and O(2), and thus no superoxide ion (O(2)(*-)) is formed. Moreover, the ESR signal of MOPDPP(*) was significantly enhanced in the presence of O(2), consistent with the assumption that the photogenerated DBT radical cation couples with O(2) to form the oxygen-adduct, which is subject to further reactions (Scheme 3) leading to the final oxygenated products. Similar results have been obtained when using 10-methylacridine hexafluorophosphate (AcrH(+)PF(6)(-), which has similar reduction potential in the ground state as MOPDPP(+)) as the sensitizer. This finding provides a possibility for the photooxidation of sulfides with dioxygen utilizing visible light (solar energy) and is also of significance in clarification of the reaction mechanism. PMID- 16471906 TI - Specific anion effects on glass electrode pH measurements of buffer solutions: bulk and surface phenomena. AB - The effect of electrolytes on pH measurements via glass electrodes is explored with solutions buffered at pH 7 (phosphate and cacodylate). Salt and buffer concentrations are varied. Direct and reverse Hofmeister effects are observed. The phenomena are significant for salt concentrations above 0.1 M and for buffer concentrations below 20 mM. Changes in measured pH show up most strongly with anions. They can be related to the usual physicochemical parameters (anion molar volumes, molar refractivity, and surface tensions) that are characteristic of Hofmeister series. They correlate strongly with anionic excess polarizabilities; this suggests the involvement of non-electrostatic, or dispersion, forces acting on ions. These forces contribute to ionic adsorption at the glass electrode surface, and to the liquid junction potential. PMID- 16471907 TI - Flux decline in nanofiltration due to adsorption of dissolved organic compounds: model prediction of time dependency. AB - Flux decline during nanofiltration of aqueous solutions containing dissolved organic compounds is mainly caused by adsorption of these compounds in the membrane pores and on the membrane surface. In this paper, flux decline is modeled by incorporating the loss of permeability due to adsorption in the Spiegler-Kedem equation. This results in a logarithmic relation between normalized flux decline and time until the adsorption equilibrium is reached and the flux reaches a constant value. In this way, the expected flux decline due to adsorption of organic compounds can be estimated. Two different parameters were used in the model: the time delay t(0), which corresponds to the time at which the adsorption process in the membrane pores and on the membrane surface sets in, and the corresponding reduction of free (pore) volume (b). Both parameters depend on the hydrophobicity of the compounds and on the feed concentration. PMID- 16471908 TI - Synthesis of novel quaternary chitosan derivatives via N-Chloroacyl-6-O triphenylmethylchitosans. AB - Various quaternary chitosan derivative structures were synthesized by reacting N chloroacyl-6-O-triphenylmethylchitosans with tertiary amines. Full substitutions were obtained from the quaternization reactions and the obtained water-soluble quaternary chitosan derivatives were thoroughly characterized with (1)H NMR, (13)C NMR, (1)H-(13)C HSQC NMR, and FT-IR. PMID- 16471909 TI - Synthesis of sugar-substituted poly(phenylenevinylene)s. AB - Sugar-containing PPVs, poly{2-[O-(2-acetamido-2-deoxy-beta-D-glucopyranosyl)]-5 methoxy-p-phenylenevinylene-alt-p-phenylenevinylene} (PPV-GlcNAc) and poly{2,5 bis-[O-(beta-d-glucopyranosyl)]-p-phenylenevinylene-alt-p-phenylenevinylene} (PPV Glc(2)), were synthesized via Heck reaction of p-divinylbenzene (DVB) with O glycosylated hydroquinones, 2,5-dibromo-4-methoxyphenyl 2-acetamido-3,4,6-tri-O acetyl-2-deoxy-beta-D-glucopyranoside (3) and 1,4-bis(O-2,3,4,6-tetra-O-acetyl beta-D-glucopyranosyl)-2,5-dibromobenzene (6), respectively. Acetyl protecting groups of the PPVs are completely removable under mild conditions (yield 69-87%). The structures were confirmed using (1)H NMR and IR spectra. Size exclusion chromatography (SEC) (eluent: DMF, polystyrene standards) measurements indicated that respective M(n) and M(w)/M(n) values of the obtained polymers are 4.49 x 10(3) and 2.3(5) (PPV-GlcNAc) and 3.86 x 10(3) and 1.3(9) (PPV-Glc(2)). These sugar-containing PPVs are soluble in water/DMF (8/2, v/v) and are recognized by Concanavalin A (Con A), D-glucose-binding protein. Blue shift of lambda(max) of the conjugated polymer backbone was confirmed when the glucose-substituted PPV interacts with Con A. Based on those binding properties, these results revealed that the obtained PPVs with pendant sugars have capabilities for detection of biological stimuli. PMID- 16471910 TI - Preparation of biopolymer fibers by electrospinning from room temperature ionic liquids. AB - Electrospinning is a versatile process used to prepare micro- and nano- sized fibers from various polymers dissolved in volatile solvents. In this report, cellulose and cellulose-heparin composite fibers are prepared from nonvolatile room temperature ionic liquid (RTIL) solvents by electrospinning. RTILs are extracted from the biopolymer fiber after the fiber formation using a cosolvent. Micron to nanometer sized, branched fibers were obtained from 10% (w/w) concentration of polysaccharide biopolymer in RTIL solution with an applied voltage of 15-20 kV. Cellulose-heparin composite fibers showed anticoagulant activity, demonstrating that the bioactivity of heparin remained unaffected even on exposure to a high voltage involved in electrospinning. PMID- 16471911 TI - Thermodynamics of binding interactions between bovine beta-lactoglobulin A and the antihypertensive peptide beta-Lg f142-148. AB - The binding capacity of bovine beta-lactoglobulin variant A (beta-Lg A) for six peptides derived from beta-Lg was evaluated using an ultrafiltration method under the following conditions: pH 6.8, 40 degrees C, and a beta-Lg A/peptide molar ratio of 1:5. Only peptides beta-Lg f102-105, f142-148, and f69-83 bound in significant amounts to beta-Lg A corresponding to 1.5, 1.1, and 0.7 mol of peptide per mole of beta-Lg A, respectively. The interaction between beta-Lg A and the antihypertensive peptide beta-Lg f142-148 was investigated further by isothermal titration calorimetry. The binding isotherms at pH 6.8 and 25 degrees C confirmed that beta-Lg f142-148 bound to beta-Lg A and that the interaction followed a sequential three-site binding model with constants of association of 2 x 10(3), 1 x 10(3), and 0.4 x 10(3) M(-1) for the first, second, and third binding sites, respectively. The enthalpy of binding was exothermic for the first and second binding sites and endothermic for the third binding site. Binding of the peptide to all three sites was spontaneous as shown by the negative free energy values. These results show for the first time that beta-Lg A can bind bioactive peptides. This potential could be exploited to transport bioactive peptides and protect them in the gastrointestinal tract following their oral administration as nutraceuticals. PMID- 16471912 TI - Introducing amine functionalities on a poly(3-hydroxybutyrate-co-3 hydroxyvalerate) surface: comparing the use of ammonia plasma treatment and ethylenediamine aminolysis. AB - Amine functionalities were introduced onto the surface of poly(3-hydroxybutyrate co-3-hydroxyvalerate) (PHBV) films by applying radio frequency ammonia plasma treatment and wet ethylenediamine treatment. The modified surfaces were characterized by X-ray photoelectron spectroscopy (XPS) for chemical composition and Raman microspectroscopy for the spatial distribution of the chemical moieties. The relative amount of amine functionalities introduced onto the PHBV surface was determined by exposing the treated films to the vapor of trifluoromethylbenzaldehyde (TFBA) prior to XPS analysis. The highest amount of amino groups on the PHBV surface could be introduced by use of ammonia plasma at short treatment times of 5 and 10 s, but no effect of plasma power within the range of 2.5-20 W was observed. Ethylenediamine treatment yielded fewer surface amino groups, and in addition an increase in crystallinity as well as degradation of PHBV was evident from Fourier transform infrared spectroscopy. Raman maps showed that the coverage of amino groups on the PHBV surfaces was patchy with large areas having no amine functionalities. PMID- 16471913 TI - Thermal diffusion of dextran in aqueous solutions in the absence and the presence of urea. AB - The Ludwig-Soret effect was studied for aqueous solutions of dextran in the temperature range 15 < T < 55 degrees C taking into account the effect of the addition of urea. In the absence of urea, the Soret coefficient S(T) changes sign; it is positive for T > 45.0 degrees C but negative for T < 45.0 degrees C. The positive sign of S(T) means that the dextran molecules migrate toward the cold side of the fluid; this behavior is typical for polymer solutions, whereas a negative sign indicates the macromolecules move toward the hot side. The addition of urea to the aqueous solution of dextran rises S(T) and reduces the inversion temperature. For 2 M urea the change in the sign of S(T) is observed at T = 29.7 degrees C and beyond that value S(T) is always positive in the studied temperature range. To rationalize these observations, it is assumed that the addition of urea leads to an opening of hydrogen bonds similar to that induced by an increase in temperature. PMID- 16471915 TI - Solution and conformational properties of wheat beta-D-glucans studied by light scattering and viscometry. AB - The solution properties of wheat beta-glucan were investigated by light scattering and viscometric methods. The hydrodynamic radius (R(h)), weight average molecular weight (M(w)), radius of gyration (R(g)), and the second virial coefficient (A(2)) of wheat beta-glucan were determined by both dynamic and static light scattering methods, whereas the critical concentrations (c) of the solution were derived from [eta] via viscometric method. The structure sensitive parameters, rho (1.52-1.62), the conformation parameter nu (0.62), and the Mark Houwink-Sakurada exponents alpha (0.78) confirmed the random coil conformation of wheat beta-glucan in 0.5 M NaOH solution. The characteristic ratio (4.97) was obtained by the random flight model, and the statistical segment length (8.83 nm) was derived from the wormlike cylinder model. It was found that the wormlike cylinder model could explain the chain stiffness better than the random flight model, which suggested an extended random coil conformation of wheat beta-glucan in 0.5 M NaOH solution. The study also revealed that the structure feature of wheat beta-glucan; that is, the higher trisaccharide-to-tetrasaccharide ratio contributed to the stiffer chain conformation compared with other cereal beta glucans. PMID- 16471914 TI - Effect of magnesium and iron on the hydration and hydrolysis of guar gum. AB - The effect that magnesium and iron have on the hydration and hydrolysis of guar gum at pH 12 was studied as a function of viscosity. It was found that small concentrations of magnesium do not affect the dissolution ratio of guar but significantly decrease hydrolysis at high temperatures. These results suggest that Mg(OH)(2) forms an adduct with the polysaccharide that prevents thermal hydrolysis of the guar. Viscosity measurements recorded in the presence of iron at pH 12 show that ferric iron inhibits hydration or dissolution of guar and may accelerate chain scission of fully hydrated guar when solutions are heated in an autoclave at 121 degrees C. PMID- 16471916 TI - Synthesis and characterization of PCL-b-PEO-b-PCL-based nanostructured and porous hydrogels. AB - Hydrogels with nanoscale structure were synthesized using amphiphilic poly(epsilon-caprolactone)-poly(ethylene oxide)-poly(epsilon-caprolactone) (PCL-b PEO-b-PCL) triblock copolymers. Small-angle X-ray scattering (SAXS) studies show that the block copolymers form 30-40 nm structures in aqueous solution and that these patterns are retained, with some increase in length scale, following electron beam cross-linking. Lamellar nanostructures were observed by SAXS and atomic force microscopy (AFM), with SAXS indicating cylindrical structure as the block lengths become more different in length. It is demonstrated through Fourier transform infrared spectroscopy (FTIR), mass loss, and differential scanning calorimetry (DSC) that the PCL can be completely removed by hydrolysis in NaOH(aq) to form porous PEO hydrogels. These hydrogels retain active functional groups following PCL removal that serve as sites for further chemical modification. PMID- 16471917 TI - Synthesis and unexpected reactivity of iron tris(bipyridine) complexes with poly(ethylene glycol) macroligands. AB - High molecular weight poly(ethylene glycol) (PEG) derivatized iron tris(bipyridine) complexes, presenting hydroxyl end groups for further modification as bioconjugates, copolymers, or cross-linking agents, were synthesized via ring-opening anionic polymerization of ethylene oxide from hydroxyl-functionalized bipyridine (bpy) initiators and subsequent chelation to iron(II). Bpy-centered PEG macroligands (bpyPEG(2)) with molecular weights ranging from 4,000 to 17,000 and low polydispersity indices (<1.1) were obtained. Chelation of the bpyPEG(2) macroligands to iron(II) sulfate was studied in aqueous solution by titration and kinetics experiments, which revealed unexpected air sensitivity compared to nonpolymeric iron tris(bipyridine) complexes. Red violet aqueous solutions of [Fe(bpyPEG(2))(3)](2+) begin to bleach within hours when exposed to air. Enhanced polymer degradation and gel formation of acrylate modified bpyPEG(2) in the presence of Fe(2+) suggest that radicals may be involved. Under argon, the chromophores are stable. Polymeric iron complexes are slower to form and faster to degrade in air with increasing bpyPEG(2) molecular weight. These studies demonstrate the influence of molecular weight in polymeric iron tris(bipyridine) complex coordination chemistry and reactivity. PMID- 16471918 TI - Effects of temperature and water content on the secondary structure of wheat gluten studied by FTIR spectroscopy. AB - The effect of temperature on gluten conditioned at the following water contents, 0%, 13%, and 47% (wet weight basis), was studied by FTIR spectroscopy over the temperature range of 25-85 degrees C. A detailed discussion of the assignment of the amide I band is given. At 0% hydration no changes in the secondary structure with temperature could be detected; spectra were consistent with a tight disordered structure with many protein-protein interactions. At 13% hydration, distinctive changes occurred in the low-frequency region of the amide I band (1,630-1,613 cm(-1)). This was attributed to changes in the beta-sheet structure. On cooling to 25 degrees C, these changes were mainly reversed. It was noted that most of the changes observed occurred above the glass transition temperature. At 47% hydration, more complex changes took place: as the temperature was raised distinct bands at 1,630 and 1,613 cm(-1) merged. However, this process was partially reversed, with recovery of both bands, on cooling. The significance of these results in relation to other changes in gluten proteins in flour and dough with temperature and water content is discussed. PMID- 16471919 TI - Spectroscopic and photothermal study of myoglobin conformational changes in the presence of sodium dodecyl sulfate. AB - Interactions between sodium dodecyl sulfate (SDS) and horse heart myoglobin (Mb) at surfactant concentrations below the critical micelle concentration have been studied using steady-state and transient absorption spectroscopies and photoacoustic calorimetry. SDS binding to Mb induces a heme transition from high spin five-coordinate to low-spin six-coordinate in met- and deoxyMb, with the distal His residue likely to be the sixth ligand. The transition is complete at an SDS concentration of approximately 350 microM and approximately 700 microM for met- and deoxyMb, respectively. DeltaG(H(2)O) and m values determined from equilibrium SDS-induced unfolding curves indicate similar stability of met- and deoxyMb toward unfolding; however, the larger m value for the deoxyMb equilibrium intermediate indicates that its structure differs from that of metMb. Results from transient absorption spectroscopy show that CO rebinding to Fe(2+)-Mb in the presence of SDS is a biphasic process with the rate constant of the first process approximately 5.5 x 10(3) s(-1), whereas the second process displays a rate similar to that for CO rebinding to native Mb (k(obs) = 7.14 x 10(2) s(-1)) at 1 mM CO. Results of photoacoustic calorimetry show that CO dissociation from deoxyMb occurs more than 10 times faster in the presence of SDS than in native Mb. These data suggest that the heme binding pocket is more solvent-exposed in the SDS-induced equilibrium intermediate relative to native Mb, which is likely due to the electrostatic and hydrophobic interactions between surfactant molecules and the protein matrix. PMID- 16471920 TI - Effects of polymer structure on the inhibition of cholera toxin by linear polypeptide-based glycopolymers. AB - A variety of important biological events are mediated by the multivalent interaction between relevant oligosaccharides and multiple saccharide receptors on lectins, toxins, and cell surfaces; a variety of glycopolymeric materials have therefore been investigated in studies aimed at manipulating these events. The synthesis of protein- and polypeptide-based glycopolymers via protein engineering and other methods offers opportunities to control both the number and the spacing of saccharides on a scaffold, as well as the conformation of the polymer backbone, and will therefore facilitate the structure-based design of polymers for inhibition of multivalent binding events. In initial studies, we have synthesized a family of galactose-functionalized glycopolymers with a poly(L glutamic acid) backbone, in which the density and linker length of the pendant carbohydrate moiety were varied. The composition of the glycopolymers was determined via (1)H NMR spectroscopy, and the impact of saccharide density and linker length, as well as the potential for these polypeptide-based glycopolymers to act as high-affinity inhibitors of the cholera toxin, has been indicated via competitive enzyme-linked immunosorbent assay and fluorescence titration experiments. The results of these studies suggest strategies for optimizing the binding of linear glycopolymers to bacterial toxins and will aid in the design of additional protein-based materials for studying the impact of multivalency, spacing, and backbone rigidity in a variety of biologically relevant binding events. PMID- 16471921 TI - Conformation and assembly of polypeptide scaffolds in templating the synthesis of silica: an example of a polylysine macromolecular "switch". AB - Although the role of polycationic macromolecules in catalyzing the synthesis of silica structures is well established, detailed understanding of the mechanisms behind the production of silica structures of controlled morphologies remains unclear. In this study, we have used both poly-L-lysine (PLL) and/or poly-D lysine (PDL) for silica synthesis to investigate mechanisms controlling inorganic morphologies. The formation of both spherical silica particles and hexagonal plates was observed. The formation of hexagonal plates was suggested, via circular dichroic spectroscopy (CD), to result from the assembly of helical polylysine molecules. We confirm that the formation of PLL helices is a prerequisite to the hexagonal silica synthesis. In addition, we present for the first time that the handedness of the helicity of the macromolecule does not affect the formation of hexagonal silica. We also show, by using two different silica precursors, that the precursor does not have a direct effect on the formation of hexagonal silica plates. Furthermore, when polylysine helices were converted to beta-sheet structure, only silica particles were obtained, thus suggesting that the adoption of a helical conformation by PLL is required for the formation of hexagonally organized silica. These results demonstrate that the change in polylysine conformation can act as a "switch" in silica structure formation and suggest the potential for controlling morphologies and structures of inorganic materials via control of the conformation of soft macromolecular templates. PMID- 16471922 TI - Deposition of pectin/poly-L-lysine multilayers with pectins of varying degrees of esterification. AB - The effect of pectin esterification on the assembly of multilayers consisting of poly-L-lysine (PLL) and pectin was studied using surface plasmon resonance (SPR), Fourier transform infrared-attenuated total reflection spectroscopy (FTIR-ATR), and a quartz crystal microbalance with dissipation monitoring (QCMD). With each layer deposited, there was a progressive increase in mass. The net charge of the multilayers was positive and increased with increasing degree of esterification of the pectin. Multilayer fabrication involved a limited fractionation of the pectin preparations, with the more highly esterified pectins having a weaker affinity for PLL. The multilayers were relatively hydrated structures with estimates of solids content in the range 10-32% w/w. The more highly esterified pectins had a tendency to form more hydrated structures, which showed a strong deswelling when PLL was added to a freshly deposited pectin layer. PMID- 16471923 TI - The Effect of monoglycerides on structural and topographical characteristics of adsorbed beta-casein films at the air-water interface. AB - The effect of monoglycerides (monopalmitin and monoolein) on the structural and topographical characteristics of beta-casein adsorbed film at the air-water interface has been analyzed by means of surface pressure (pi)-area (A) isotherms and Brewster angle microscopy (BAM). At surface pressures lower than that for the beta-casein collapse (pi(c)(beta-casein)), attractive interactions between beta casein and monoglycerides were observed. At higher surface pressures, the collapsed beta-casein is partially displaced from the interface by monoglycerides. However, beta-casein displacement by monoglycerides is not quantitative at the monoglyceride concentrations studied in this work. From the results derived from these experiments, we have concluded that interactions, miscibility, and displacement of proteins by monoglycerides in adsorbed mixed monolayers at the air-water interface depend on the particular protein monoglyceride system, the interactions between film-forming components being higher for adsorbed than for spread films. The adsorbed films are more segregated than spread films, and both collapsed protein domains and monoglyceride domains in adsorbed films are smaller than for spread films. PMID- 16471924 TI - Nanoparticles of beta-cyclodextrin esters obtained by self-assembling of biotransesterified beta-cyclodextrins. AB - The synthesis of decanoate beta-cyclodextrin esters (beta-CDd) and hexanoate beta cyclodextrin esters (beta-CDh) was biocatalyzed by thermolysin from native beta cyclodextrin (beta-CD) and vinyl hexanoate or vinyl decanoate used as acyl donors. The products were chemically characterized by infrared, NMR, and mass spectrometry. Both beta-CDd and beta-CDh esters were identified as a mixture of beta-CD preferentially substituted on the C2 position by the corresponding acyl chain. The degree of substitution varied from 2 to 7 for beta-CDd and from 4 to 8 for beta-CDh. The ability of beta-CD esters to self-organize into nanoparticles was tested using a nanoprecipitation technique in various solvents. The mean size diameter and polydispersity measured by quasi-elastic light scattering were dramatically affected by the nature of solvent (acetone, ethanol, or tetrahydrofuran) used in the nanoprecipitation technique. When directly observed using cryo-transmission electron microscopy, beta-CDh appeared as uniformly dense nanospheres, whereas beta-CDd exhibited a multilamellar onion-like organization. A structural model was rationalized for the beta-CDd nanoparticles. PMID- 16471925 TI - Fluorescence recovery after photobleaching as a probe of diffusion in starch systems. AB - The diffusion coefficients of dextran probes of various molecular weights in starch solutions over a wide concentration range were carried out using fluorescent recovery after photobleaching (FRAP), combined with a confocal microscope and tracer probe diffusion. The technique is simple to implement and can be carried out using increasingly common microscopy apparatus, giving access to a wide variety of new structural and kinetic information. The data can be rationalized in terms of the effects of probe molecular weight and on matrix starch concentration and structure. This provides a new tool to investigate the behavior of systems where starch is an ingredient that contributes to the processing and textural properties of food. PMID- 16471926 TI - Thermoplastic starch-waxy maize starch nanocrystals nanocomposites. AB - Waxy maize starch nanocrystals obtained by hydrolysis of native granules were used as a reinforcing agent in a thermoplastic waxy maize starch matrix plasticized with glycerol. Compared to our previous studies on starch nanocrystals reinforced natural rubber (NR) [Macromolecules 2005, 38, 3783; 2005, 38, 9161], the present system presents two particularities: (i) thermoplastic starch is a polar matrix, contrarily to NR, and (ii) the chemical structures of the matrix and the filler are similar. The influence of the glycerol content, filler content, and aging on the reinforcing properties of waxy maize starch nanocrystals (tensile tests, DMA) and crystalline structure (X-ray diffraction) of materials were studied. It was shown that the reinforcing effect of starch nanocrystals can be attributed to strong filler/filler and filler/matrix interactions due to the establishment of hydrogen bonding. The presence of starch nanocrystals leads to a slowing down of the recrystallization of the matrix during aging in humid atmosphere. PMID- 16471927 TI - Detection of polyclonal antibody against any area of the protein-antigen using immobilized protein-antigens: the critical role of the immobilization protocol. AB - Antigens immobilized on solid supports may be used to detect or purify their corresponding antibodies (Ab) from serum. Direct immobilization of antigens on support surfaces (through short spacer arms) may promote interesting stabilizing effects on the immobilized antigen. However, the proximity of the support may prevent the interaction of some fractions of polyclonal Ab with some regions of the antigen (those placed in close contact with the support surface). Horseradish peroxidase (HRP) was immobilized on agarose by different protocols of multipoint covalent immobilization involving different regions of the antigen surface. Glyoxyl-agarose, BrCN-agarose, and glutaraldehyde-agarose were used as activated supports. Each HRP-immobilized preparation was much more stable than the soluble enzyme, but it was only able to adsorb up to 60-70% of a mixture of polyclonal anti-HRP antibodies. On the other hand, HRP was also immobilized on agarose through a very long, flexible, and hydrophilic spacer arm (dextran). This immobilized HRP was hardly stabilized, but it was able to adsorb 100% of the polyclonal anti-HRP. The absence of steric hindrances seems to play a critical role favoring the complete recognition of all classes of polyclonal Ab. Another solution to achieve a complete adsorption of polyclonal Ab on immobilized stabilized antigens has been also reached by using a mixture of the differently immobilized and stabilized HRP-agarose preparations. In this case, an improved storage and operational stabilities of the immobilized antigens can be combined with the complete adsorption of any class of antibody. PMID- 16471928 TI - Hydroxylated poly(N-isopropylacrylamide) as functional thermoresponsive materials. AB - In this study, we developed a poly(N-isopropylacrylamide)-based thermoresponsive polymeric material with a high content of hydroxyl groups. We newly designed the functional monomer, N-(2-hydroxyisopropyl)acrylamide (HIPAAm), considering maintaining the continuous and repeated structure of the isopropylamide group after copolymerization and the monomer reactivity ratios. The thermoresponsive polymer was derived by conventional radical copolymerization of HIPAAm with N isopropylacrylamide (NIPAAm) in high yield. Estimation of monomer reactivity ratios, r(1) and r(2), supported the almost random sequence of the comonomers. The obtained copolymers showed a very sensitive phase transition and/or separation in response to temperature in aqueous media although they have many hydrophilic parts, and their thermoresponsive behavior was not affected by the pH. Furthermore, the cloud points of these copolymers closely depended on the HIPAAm content and could be easily controlled by adding salts. HIPAAm is expected to regulate the phase transition and/or separation temperature of the NIPAAm based copolymers while maintaining their desirable sensitive thermoresponse. Differential scanning calorimetric analysis showed that dehydration of the polymer chains occurring in phase transition became incomplete with increasing HIPAAm content. Moreover, it was found that poly(NIPAAm-co-HIPAAm) having a high content of the HIPAAm unit showed liquid-liquid phase separation involving coacervation. The sizes of the coacervate droplets were relatively monodisperse and very minimal. Poly(NIPAAm-co-HIPAAm) is valuable for use in biomedical fields such as bioseparation. PMID- 16471929 TI - Purification and functional analysis of a 40 kD protein extracted from the Strombus decorus persicus mollusk shells. AB - A 40 kD protein has been extracted from the biomineral matrix of the calcium carbonate gastropod shell of Strombus decorus persicus. The protein was isolated by decalcification and ion exchange HPLC. We have named this protein ACLS40, i.e., aragonite crossed-lamellar structure protein. A partial sequence of the isolated ACLS40 and amino acid analysis both indicate that it does not belong to the family of very acidic proteins, i.e., rich in aspartic and glutamic residues. The shell-extracted protein shows the ability to stabilize calcium carbonate in vitro, in the form of thermodynamically unstable vaterite polymorph, and to inhibit the growth of calcite. PMID- 16471930 TI - Dextran functionalized surfaces via reductive amination: morphology, wetting, and adhesion. AB - Dextran surface grafting density was systematically varied via a two-step process involving SiO(2) amination by aminopropyltriethoxy silane (APTES) followed by oxidized dextran (M(w) = 110 kDa) chemisorption. Dextran oxidation kinetics with sodium metaperiodate (NaIO(4)) were quantified by (1)H NMR and pH measurements. Aldehyde group formation increased with increasing oxidation time. For 0.5 h oxidation time, dried film ellipsometric thickness was constant for solution concentrations ranging from 1 to 4 mg/mL. Dextran layers with the lowest grafting density wetted fastest and displayed the lowest contact angle (theta(APTES) > theta(1 h) > theta(2,4 h) > theta(0.5 h)). Under aqueous conditions, AFM force versus displacement measurements on 0.5 and 4.0 h surfaces exhibited a single displacement jump upon retraction. The 1.0 and 2.0 h surfaces showed two jumps consistent with two populations of chains, namely, loosely and strongly bound dextran. Overall, film morphology and wetting behavior were relatively invariant with grafting density, confirming the method's robustness for preparing biomimetic coatings with consistent properties. PMID- 16471931 TI - Aggregation of antifreeze glycoprotein fraction 8 and its effect on antifreeze activity. AB - Antifreeze glycoproteins (AFGPs) have many potential applications ranging from the cryopreservation and hypothermic storage of tissues and organs to the preservation of various frozen food products. Since supplying native AFGP for these applications is a labor-intensive and costly process, the rational design and synthesis of functional AFGP analogues is a very attractive alternative. While structure-function studies have implicated specific structural motifs as essential for antifreeze activity in AFGP, the relationship between solution conformation and antifreeze activity is poorly understood. Toward this end, we have analyzed AFGP8 in aqueous solutions using dynamic light scattering (DLS) and circular dichroism (CD). Our results indicate that AFGP8 forms discrete aggregates in solution. These aggregates are predominantly composed of dimers that form at solution concentrations greater than 20 mM. CD spectroscopy indicates that the preferred solution conformation of AFGP8 is consistent with that of random coil. However, significant beta-sheet and alpha-helix character is observed in more concentrated solutions, indicating that these glycopeptides are highly flexible in solution. Aggregation appears to have a minimal effect on the overall solution conformation. Thermal hysteresis (TH) activity of the aggregated solutions is much higher than that of less concentrated solutions that do not form aggregates. While cooperative functioning between lower and higher molecular weight AFGPs has been reported, this is the first instance where cooperative functioning in lower molecular weight AFGPs has been observed. PMID- 16471933 TI - pH controlled permeability of lipid/protein biomimetic microcapsules. AB - The permeability of lipid and protein microcapsules fabricated by alternating adsorption of human serum albumin (HSA) and L-alpha-dimyristoylphosphatidic acid (DMPA) on a template and subsequent removal of the core is studied as a function of pH value and supplementary layers. The capsules were permeable for macromolecules (FITC-labeled dextran, M(w) 40 kDa) at pH < 4.8 and impermeable at pH > 7.4. The assembly of supplementary DMPA bilayers rendered the capsules impermeable for small hydrophilic molecules such as 6-carboxyfluorescein (6-CF). Hence DMPA/HSA capsules can be resealed after fabrication by supplementary layers. This provides the opportunity of applying such biomimetic membrane capsules as drug carriers or model systems to study biological processes at membranes. PMID- 16471932 TI - PAMAM dendrimer-based multifunctional conjugate for cancer therapy: synthesis, characterization, and functionality. AB - Poly(amidoamine) (PAMAM) dendrimer-based multifunctional cancer therapeutic conjugates have been designed and synthesized. The primary amino groups on the surface of the generation 5 (G5) PAMAM dendrimer were neutralized through partial acetylation, providing enhanced solubility of the dendrimer (in conjugation of FITC (fluorescein isothiocyanate)) and preventing nonspecific targeting interactions (in vitro and in vivo) during delivery. The functional molecules fluorescein isothiocyanate (FITC, an imaging agent), folic acid (FA, targets overexpressed folate receptors on specific cancer cells), and paclitaxel (taxol, a chemotherapeutic drug) were conjugated to the remaining nonacetylated primary amino groups. The appropriate control dendrimer conjugates have been synthesized as well. Characterization of the G5 PAMAM dendrimer and its nanosize conjugates, including the molecular weight and number of primary amine groups, has been determined by multiple analytical methods such as gel permeation chromatography (GPC), nuclear magnetic resonance spectroscopy (NMR), potentiometric titration, high-performance liquid chromatography (HPLC), and UV spectroscopy. These multifunctional dendrimer conjugates have been tested in vitro for targeted delivery of chemotherapeutic and imaging agents to specific cancer cells. We present here the synthesis, characterization, and functionality of these dendrimer conjugates. PMID- 16471934 TI - Biomimetic synthesis of water-soluble conducting copolymers/homopolymers of pyrrole and 3,4-ethylenedioxythiophene. AB - A novel biomimetic route for the synthesis of electrically conducting homopolymers/copolymers of pyrrole and 3,4-ethylenedioxythiophene (EDOT) in the presence of a polyelectrolyte, such as polystyrene sulfonate (SPS), is presented. A poly(ethylene glycol)-modified hematin (PEG-hematin) was used to catalyze the homopolymerization of pyrrole and EDOT as well as copolymerization of EDOT and pyrrole in the presence of SPS to yield homopolymers of polypyrrole/SPS and PEDOT/SPS as well as a polypyrrole-co-poly(3,4-ethylenedioxythiophene)/SPS complex. Spectroscopic characterization [UV-visible, Fourier transform infrared (FTIR), and X-ray photoelectron spectroscopy (XPS)], thermal analysis, (TGA), and electrical conductivity studies for these complexes indicated the presence of a stable and electrically conductive form of these polymers. Furthermore, the presence of SPS that serves as a charge-compensating dopant in this complex provides a unique combination of properties such as processability and water solubility. PMID- 16471935 TI - Synthesis and characterization of RGD peptide grafted poly(ethylene glycol)-b poly(L-lactide)-b-poly(L-glutamic acid) triblock copolymer. AB - Advances in tissue engineering require biofunctional scaffolds that can provide not only physical support for cells but also chemical and biological cues needed in forming functional tissues. To achieve this goal, a novel RGD peptide grafted poly(ethylene glycol)-b-poly(L-lactide)-b-poly(L-glutamic acid) (PEG-PLA-PGL/RGD) was synthesized in four steps (1) to prepare diblock copolymer PEG-PLA-OH and to convert its -OH end group into -NH(2) (to obtain PEG-PLA-NH(2)), (2) to prepare triblock copolymer PEG-PLA-PBGL by ring-opening polymerization of NCA (N carboxyanhydride) derived from benzyl glutamate with diblock copolymer PEG-PLA NH(2) as macroinitiator, (3) to remove the protective benzyl groups by catalytic hydrogenation of PEG-PLA-PBGL to obtain PEG-PLA-PGL, and (4) to react RGD (arginine-glycine-(aspartic amide)) with the carboxyl groups of the PEG-PLA-PGL. The structures of PEG-PLA-PGL/RGD and its precursors were confirmed by (1)H NMR, FT-IR, amino acid analysis, and XPS analysis. Addition of 5 wt % PEG-PLA-PGL/RGD into a PLGA matrix significantly improved the surface wettability of the blend films and the adhesion and proliferation behavior of human chondrocytes and 3T3 cells on the blend films. Therefore, the novel RGD-grafted triblock copolymer is expected to find application in cell or tissue engineering. PMID- 16471936 TI - Nanoscale anionic macromolecules can inhibit cellular uptake of differentially oxidized LDL. AB - Nanoscale particles could be synthetically designed to potentially intervene in lipoprotein matrix retention and lipoprotein uptake in cells, processes central to atherosclerosis. We recently reported on lipoprotein interactions of nanoscale micelles self-assembled from amphiphilic scorpion-like macromolecules based on a lauryl chloride-mucic acid hydrophobic backbone and poly(ethylene glycol) shell. These micelles can be engineered to present varying levels of anionic chemistry, a key mechanism to induce differential retentivity of low-density lipoproteins (LDL) (Chnari, E.; Lari, H. B.; Tian, L.; Uhrich, K. E.; Moghe, P. V. Biomaterials 2005, 26, 3749). In this study, we examined the cellular interactions and the ability of carboxylate-terminated nanoparticles to modulate cellular uptake of differentially oxidized LDL. The nanoparticles were found to be highly biocompatible with cultured IC21 macrophages at all concentrations examined. When the nanoparticles as well as LDL were incubated with the cells over 24 h, a marked reduction in cellular uptake of LDL was observed in a nanoparticle concentration-dependent manner. Intermediate concentrations of nanoparticles (10(-6) M) elicited the most charge-specific reduction in uptake, as indicated by the difference in uptake due to anionic and uncharged nanoparticles. At these concentrations, anionic nanoparticles reduced LDL uptake for all degrees of oxidation (no oxidation, mild, high) of LDL, albeit with qualitative differences in the effects. The anionic nanoparticles were particularly effective at reducing the very high levels of uptake of the most oxidized level of LDL. Since complexation of LDL with anionic nanoparticles is reduced at higher degrees of LDL oxidation, our results suggest that anionic nanoparticles interfere in highly oxidized (hox) LDL uptake, likely by targeting cellular/receptor uptake mechanism, but control unoxidized LDL uptake by mechanisms related to direct LDL-nanoparticle complexation. Thus, anionically functionalized nanoparticles can modulate the otherwise unregulated internalization of differentially oxidized LDL. PMID- 16471937 TI - Tethering functional ligands onto shell of ultrasound active polymeric microbubbles. AB - Hollow (air-filled) microparticles, i.e., microbubbles, provide a promising novel vehicle for both local delivery of therapeutic agents and simultaneous diagnostic ultrasound echo investigations. In this paper, we describe the synthetic routes for decorating the polymeric shell of a poly(vinyl alcohol)-based microbubble with low and high molecular weight ligands with pharmacological relevance. Investigations on physical properties of microbubbles and surface chemical coupling with different cargo molecules such as L-cysteine, L-lysine, poly(L lysine), chitosan, and beta-cyclodextrin were carried out by CD and NMR spectroscopies, confocal laser scanning microscopy, and microcalorimetry. The in vitro cytotoxicity and biocompatibility of the polymer microbubbles have been also determined toward different cell lines. The results are discussed in terms of the features shown by this device, i.e., injectability, long shelf life, ease of preparation, biocompatibility, loading and cargo capacities, and functional properties. PMID- 16471938 TI - Comparison of the hydrolytic degradation and deformation properties of a PLLA lauric acid based family of biomaterials. AB - Addition of lauric acid to PLLA results in a significantly increased rate of hydrolytic degradation, with the time-to-loss of tensile strength directly related to the concentration of lauric acid. In this study, the hydrolytic degradation profiles of four materials were studied: amorphous PLLA, amorphous PLLA containing 1.8 wt % lauric acid, amorphous PLLA containing 4.5 wt % lauric acid, and pre-crystallized PLLA containing 1.8 wt % lauric acid. Hydrolytic degradation was monitored through mass profiles, molecular weight profiles, crystallinity and the development of mechanical properties and deformation mechanisms (through simultaneous small-angle X-ray scattering and tensile testing), and a "phase diagram" of properties suggested. The key factor in determining the development of properties was found to be the time at which crystallization occurred in relation to the loss of molecular weight, with the two factors most affecting this being the lauric acid content and the pre degradation annealing treatment. PMID- 16471939 TI - Thermodynamic confinement and alpha-helix persistence length in poly(gamma-benzyl L-glutamate)-b-poly(dimethyl siloxane)-b-poly(gamma-benzyl-L-glutamate) triblock copolymers. AB - The structure and the associated dynamics of a series of poly(gamma-benzyl-L glutamate)-b-poly(dimethyl siloxane)-b-poly(gamma-benzyl-L-glutamate) (PBLG-b PDMS-b-PBLG) triblock copolymers were investigated using small- and wide-angle X ray scattering, NMR, transmission electron microscopy, and dielectric spectroscopy, respectively. The structural analysis revealed phase separation in the case of the longer blocks with defected alpha-helical segments embedded within the block copolymer nanodomains. The alpha-helical persistence length was found to depend on the degree of segregation; thermodynamic confinement and chain stretching results in the partial annihilation of helical defects. PMID- 16471940 TI - Conformational study of silk-like peptides containing the calcium-binding sequence from calbindin D9k using 13C CP/MAS NMR spectroscopy. AB - The calcium-binding sites of calbindin D(9k) have a helix-loop-helix motif. In this study, the helix motifs were replaced by several Ala-Gly repeating regions designed on the basis of the primary sequences of several silk fibroins. The synthesized peptides were treated with several organic solvents to modify the secondary structure of the Ala-Gly repeating regions. The local structures of the Ala-Gly repeating regions, as well as the calcium-binding motif, D(9k)-loop (D(9k)L), were determined by (13)C CP/MAS NMR spectroscopy. In the four peptides containing D(9k)L synthesized, the poly(Ala) domains retain the ability to undergo a conformational transition from alpha-helical to beta-sheet in (A)(12) D(9k)L despite the presence of the D(9k)L domain at the center of the peptide molecule, but the presence of this domain in the other model peptides synthesized has a marked effect on the conformation of the added silk-like domains. The results showed that the structures of the Ala-Gly repeating regions can be controlled by the choice of both the organic solvent and the amino acid sequence of the Ala-Gly repeating regions without disrupting the secondary structure of D(9k)L suggesting that it may retain its ability to bind calcium ions. PMID- 16471941 TI - Biomimetic nanofibrous scaffolds: preparation and characterization of PGA/chitin blend nanofibers. AB - Electrospinning of poly(glycolic acid) (PGA)/chitin blend solutions in 1,1,1,3,3,3-hexafluoro-2-propanol was investigated to fabricate biodegradable and biomimetic nanostructured scaffolds for tissue engineering. The morphology of the electrospun PGA/chitin blend nanofibers was investigated with a field emission scanning electron microscope. The PGA/chitin blend fibers have average diameters of around 140 nm, and their diameters have a distribution in the range 50-350 nm. The miscibility of PGA/chitin blend fibers was examined by differential scanning calorimetry. The PGA and chitin were immiscible in the as-spun nanofibrous structure. An in vitro degradation study of PGA/chitin blend nanofibers was conducted in phosphate-buffered saline, pH 7.2. It was found that the hydrolytic cleavage of PGA in the blend nanofibers was accelerated by the coexistence of hydrophilic chitin. To assay the cytocompatability and cell behavior on the PGA/chitin blend nanofibrous scaffolds, cell attachment and spreading of normal human epidermal fibroblasts seeded on the scaffolds were studied. Our results indicate that the PGA/chitin blend nanofibrous matrix, particularly the one that contained 25% PGA and 75% chitin with bovine serum albumin coating, could be a good candidate for tissue engineering scaffolds, because it has an excellent cell attachment and spreading for normal human fibroblasts. PMID- 16471942 TI - A transition from cellulose swelling to cellulose dissolution by o-phosphoric acid: evidence from enzymatic hydrolysis and supramolecular structure. PMID- 16471943 TI - Coordination complexes with cis-TCNE radical anion ligands. models of M[TCNE]2 magnets. AB - The synthesis and characterization of two manganese(II) complexes formally each featuring two cis-tetracyanoethylenide radical anionic ligands (TCNE*/-) are reported. In each case, tris(pyrazol-1-ylmethyl)amine serves as a capping ligand, blocking three facial coordination sites. Crystal structures show that the two TCNE anions in each molecule exhibit an intramolecular stacking interaction that forms what can be considered a coordinated (TCNE2)2- moiety. These molecules are presumed to be structural models of some of the local bonding in the family of amorphous, ferrimagnetic, M[TCNE]2.y(solvent) coordination polymer magnets. Magnetic measurements indicate that the (TCNE2)2- bridge is diamagnetic and not a good mediator of magnetic exchange, a result that might explain the observed lower ordering temperatures in some of the polymer magnets. PMID- 16471945 TI - New disordering mode for TFSI- anions: the nonequilibrium, plastic crystalline structure of Et4NTFSI. AB - A new TFSI- anion disordering mode has been discovered in a supercooled plastic crystalline phase of Et4NTFSI, which may, in part, account for the low melting points of TFSI- salts with organic cations, thereby forming ionic liquids, and the intriguing properties of LiTFSI for lithium battery applications. PMID- 16471944 TI - Geometric preferences in iron(II) and zinc(II) model complexes of peptide deformylase. AB - A combination of experimental and theoretical studies on (N,S(thiolate))M(II) formate complexes (M = Fe, Zn) suggests a rationale for the metal ion dependence of peptide deformylase. PMID- 16471946 TI - Effect of the transition metal on the synthesis of quaternary sulfides MPb8In17S34 (M = Cu, Ag, Au). AB - Quaternary sulfides MPb8In17S34 (M = Cu, Ag, Au) were synthesized at 1123 K from their elements in stoichiometric ratios. These compounds crystallize in monoclinic space group P2(1)/m. The crystal structures feature combinations of 2[InS2] (NaCl111-type) and [MPb2In3S6] (NaCl100-type) slabs. These compounds are semiconductors with band gaps near 1.3 eV. PMID- 16471947 TI - A new type of luminescent alkynyl Au4Cu2 cluster. AB - Changes in the optical properties of an alkynyldigold(I) complex upon reaction with Cu(I) are associated with a complicated structural change to form an unusual Au4Cu2 cluster with metallophilic interactions as well as pi-alkyne coordination. PMID- 16471948 TI - Metal clusters as ligands. Substitution of fe ions in Fe/Mo/S clusters by thiophilic CuI ions. AB - The reactivity of Fe/S and Fe/Mo/S clusters, similar or analogous to those occurring in biological systems, with thiophilic metal ions has not been explored. In this Communication, we demonstrate that synthesis of heteropolynuclear clusters with different coordination geometries for different metals at different sites is possible by metal substitution or by metal addition reactions. The two clusters we report herein ([(Cl4 cat)2Mo2Cu5Fe4S9(PnPr3)7(SPnPr3)2]PF6 and [(Cl4-cat)2Mo2Cu6Fe4S10(PnPr3)8]) contain Fe, Mo, and Cu, which display pseudotetrahedral, pseudooctahedral, and pseudotrigonal geometries, respectively. The synthesis of these clusters is achieved by the addition of appropriate amounts of [Cu(CH3CN)4]+ to [(Cl4 cat)2Mo2Fe8(PnPr3)6]. The formation of the different products is temperature- and solvent-dependent. The Cu(I) units incorporated into the metal cluster framework, either bind to available lone pairs of the already bridging S ligands or displace the less thiophilic Fe atoms. Among the essential features of these new molecules are recognizable Fe/S fragments including an Fe6S9 core in the first cluster and the pentlandite Fe4Cu4S6 core in the second cluster. PMID- 16471949 TI - DFT analysis of interligand vibrations in a hydroperoxo complex of cobalt bleomycin. AB - Density functional theory (DFT) has been applied to the analysis of interligand vibrations in two chiral isomers of hydroperoxo complex of cobalt bleomycin (BLM Co(III)-OOH, BLM = bleomycin). The DFT-based normal coordinate analysis reveals that 16O/18O isotope-sensitive modes associated with the Co-OOH moiety uniquely reflect the chiral organization of ligands around the cobalt atom. This study provides an independent probe of cobalt chirality coordinated to BLM and shows that interligand modes associated with the Co-OOH moiety could be used as a structural marker of the chiral isomers. PMID- 16471950 TI - Sulfur stable isotope distribution of polysulfide anions in an (NH4)2Sn aqueous solution. AB - Methylation of polysulfides [(NH4)2Sn)] by reaction with CF3SO3CH3 followed by separation of the produced dimethylpolysulfides by liquid chromatography and subsequent highly accurate stable isotope analysis by a continuous-flow isotope ratio mass spectrometer shows that polysulfide anions in an aqueous solution exchange isotopes with the other sulfur species in the system. It demonstrates for the first time that polysulfide anions are 34S-enriched in equilibrium relative to total sulfur as a function of their sulfur chain length. PMID- 16471951 TI - Two new borates containing the first examples of large isolated polyborate anions: chain [B7O9(OH)5]2- and ring [B14O20(OH)6]4-. AB - Two novel hydrated borates were synthesized under mild solvothermal conditions. One is hydrated rubidium borate, Rb2B7O9(OH)5, which contains the first example of the isolated chain heptaborate anion, [B7O9(OH)5]2-. The other is hydrated diethylenetriamine borate, [(C4H15N3)]2[B14O20(OH)6], which contains the first example of the largest isolated ring tetradecaborate anion, [B14O20(OH)6]4-. PMID- 16471952 TI - On the composition and crystal structure of the new quaternary hydride phase Li4BN3H10. AB - X-ray data on single crystals of the quaternary metal hydride near the composition LiB(0.33)N(0.67)H(2.67), previously identified as "Li3BN2H8", reveal that its true composition is Li4BN3H10. The structure has body-centered-cubic symmetry [space group I2(1)3, cell parameter a = 10.679(1)-10.672(1) Angstroms] and contains an ordered arrangement of BH4- and NH2- anions in the molar ratio 1:3. The borohydride anion has an almost ideal tetrahedral geometry (angleH-B-H approximately 108-114 degrees), while the amide anion has a nearly tetrahedral bond angle (angleH-N-H approximately 106 degrees). Three symmetry-independent Li atom sites are surrounded by BH4- and NH2- anions in various distorted tetrahedral configurations, one by two B and two N atoms, another by four N atoms, and the third by one B and three N atoms. The Li configuration around B is nearly tetrahedral, while that around N resembles a distorted saddlelike configuration, similar to those in LiBH4 and LiNH2, respectively. PMID- 16471953 TI - UF3(H2O)(C2O4)0.5: a fluorooxalate of tetravalent uranium with a three dimensional framework structure. AB - A new uranium(IV) fluorooxalate, UF3(H2O)(C2O4)0.5, has been synthesized by a hydrothermal method and structurally characterized by single-crystal X-ray diffraction, infrared spectroscopy, and thermogravimetric analysis. The structure consists of two-dimensional layers of corner- and edge-sharing tricapped trigonal prisms with the composition UF(4/2)F(2/2)O3 linked by bisbidentate oxalate ligands to form a three-dimensional framework. Magnetic susceptibilities were measured to confirm the tetravalent state of uranium. Crystal data: monoclinic, space group C2/c, a = 17.246(3) Angstroms, b = 6.088(1) Angstroms, c = 8.589(2) Angstroms, beta = 95.43(3) degrees, and Z = 8. PMID- 16471954 TI - Layer-by-layer deposition of bimetallic nanoshells on functionalized polystyrene beads. AB - Bimetallic nanoshells on functionalized polystyrene beads have been fabricated through a layer-by-layer deposition technique exploiting electrostatic interaction. The synthesis has been achieved through the immobilization and successive reduction of the corresponding precursor ions. It has been shown that the thickness of the shell can be controlled by a number of cyclic depositions of respective metals onto the surface of the polystyrene beads. PMID- 16471955 TI - Robust non-interpenetrating coordination frameworks from new shape-persistent building blocks. AB - New shape-persistent ligands derived from triptycene were synthesized, and reaction with copper iodide results in the assembly of non-interpenetrating coordination frameworks with hydrophobic void spaces. These structures are thermally stable and display reversible solvent adsorption, and 1H NMR experiments show that they can be used to remove benzene from water. PMID- 16471956 TI - Asymmetric transformation of a double-stranded, dicopper(I) helicate containing achiral bis(bidentate) Schiff bases. AB - Reactions of the bis(bidentate) Schiff-bases N,N'-bis(6-alkyl-2 pyridylmethylene)ethane-1,2-diamine (where alkyl = H, Me, iPr) (L) with tetrakis(acetonitrile)copper(I) hexafluorophosphate and silver(I) hexafluorophosphate afforded, respectively, the double-stranded, dinuclear metal helicates [T-4-(R,R)]-(+/-)-[M2L2](PF6)2 (M = Cu, Ag). The helicates were characterized by 1H and 13C NMR spectroscopy, conductivity, microanalysis, and single-crystal X-ray structure determinations on selected compounds. Intermolecular ligand exchange and intramolecular inversion rates for the complexes were investigated by 1H NMR spectroscopy. Reversible intermolecular ligand exchange between two differently substituted helicates followed first order kinetics. The rate constants (k) and corresponding half-lives (t(1/2)) for ligand exchange for the dicopper(I) helicates were k = (1.6-1.8) x 10(-6) s(-1) (t(1/2) = 110-120 h) in acetone-d6, k = 4.9 x 10(-6) s(-1) (t(1/2) = 40 h) in dichloromethane-d2, and k > 2 x 10(-3) s(-1) (t(1/2) < 5 min) in acetonitrile-d3. Ligand exchange for the disilver(I) helicates occurred with k > 2 x 10(-3) s(-1) (t(1/2) < 5 min). Racemization of the dicopper(I) helicate by an intramolecular mechanism was investigated by determination of the coalescence temperature for the diastereotopic isopropyl-Me groups in the appropriate complex, and DeltaG() >> 76 kJ mol(-1) was calculated for the process in acetone-d6, nitromethane-d3, and dichloromethane-d2 with DeltaG() = 75 kJ mol(-1) in acetonitrile-d3. Complete anion exchange of the hexafluorophosphate salt of a dicopper(I) helicate with the enantiomerically pure Delta-(-)-tris(catecholato)arsenate(V) ([As(cat)3]-) in the presence of Dabco gave the two diastereomers (R,R)-[Cu2L2][Delta-(-)-[As(cat)3]]2 and (S,S)-[Cu2L2][Delta-(-)-[As(cat)3]]2 in up to 54% diastereomeric excess, as determined by (1)H NMR spectroscopy. The diastereomerically and enantiomerically pure salt (R,R)-[Cu(2)L2][Delta-(-)-[As(cat)3]]2 crystallized from the solution in a typical second-order asymmetric transformation. The asymmetric transformation of the dicopper(I) helicate is the first synthesis of a diastereomerically and enantiomerically pure dicopper(I) helicate containing achiral ligands. PMID- 16471957 TI - Synthesis and electrochemistry of undeca-substituted metallo-benzoylbiliverdins. AB - The high-yield preparation of metallo-benzoylbiliverdins 9, 10, and 11 from either oxidation of dodeca-substituted porphyrin 6 in the presence of NaNO2/TFA and air followed by metalation or by reaction of the Ni(II) or Cu(II) complexes of 6 with m-chloroperoxybenzoic acid in pyridine under air is reported. The X-ray structures of complexes 9 and 10 and the electrochemistry and spectroelectrochemistry of metallo-benzoylbiliverdins 9-11 are presented and discussed. PMID- 16471958 TI - Reduction of vanadium(V) by L-ascorbic acid at low and neutral pH: kinetic, mechanistic, and spectroscopic characterization. AB - L-Ascorbic acid interacts with vanadium(V) over the pH range of 0.4-7.0 to form three different coordination complexes. Both inner- and outer-sphere electron transfer pathways are proposed to form vanadium(IV) complexes with L-ascorbate or dehydroascorbate, respectively. Effects of the pH on the coordination of L ascorbic acid to the vanadium(V) center were observed and are presumably related to the speciation of the vanadium(V) ion. Three vanadium(IV) complexes were observed using ambient-temperature electron paramagnetic resonance spectroscopy. Two of these complexes are proposed to be vanadium(IV) L-ascorbate complexes, and one is consistent with a vanadium(IV) dehydroascorbic acid complex proposed earlier. These reduction reactions will occur under physiological conditions and could be important to the reduction of vanadium(V)-containing coordination complexes used as insulin-enhancing agents for treatment of diabetes. PMID- 16471959 TI - A density functional study of uranyl monocarboxylates. AB - We studied uranium(VI) monocarboxylate complexes by a relativistic density functional method using simple carboxylic acids as ligands, i.e. [UO2(OOCR)]+ (R = H, CH3, CH2CH3). These complexes exist in aqueous solution and, for R = CH3 and CH2CH3, may also be considered as models of uranyl complexated by humic substances. We investigated mono- and bidentate coordination modes. Short-range solvent effects were accounted for explicitly via aqua ligands of the first hydration shell and long-range electrostatic interactions were described via a polarizable continuum model. The calculated results for the uranyl U=O bond, the bond to aqua ligands, and the averaged uranium distances to equatorial oxygen atoms, U-Oeq, agreed quite well with EXAFS-derived interatomic distances. However, the uranyl-carboxylate bond was calculated to be notably shorter than the experimentally determined value. Experimental differences between mono- and bidentate coordination, obtained mainly from crystal structures, were qualitatively reproduced for the U-C distance but not for the average bond length, U-Oeq. We discuss these discrepancies between calculated and experimental results in some detail and suggest changes in the coordination number rather than variations of the coordination geometry as the main source of the experimentally observed variation of the U-Oeq distance. PMID- 16471960 TI - Density functional study of the O2 binding to [CuI(TPAR)]+ (TPA = tris(2 pyridylmethyl)amine) in THF and EtCN. AB - Density functional theory using the B3LYP hybrid functional has been employed to study the formation of [Cu(II)(TPA(H))(O2-)]+ and [Cu(II)(TPA(MeO))(O2-)]+ (TPA = tris(2-pyridylmethyl)amine) in two different solvents, THF and EtCN. The thermodynamics of solvent coordination as well as that of the overall reactions with O2 has been computed. The formations of [Cu(II)(TPA(H))(O2-)]+ in THF and of [Cu(II)(TPA(MeO))(O2-)]+ in both THF and EtCN are found to be initiated from the [Cu(I)(TPA(R))]+ species, that is, the Cu complex possessing an empty coordination site. In contrast, the formation of [Cu(II)(TPA(H))(O2-)]+ in EtCN is found to be initiated from the [Cu(I)(TPA(H))(EtCN)]+ species, that is, one solvent molecule being coordinated to Cu(I). In general, good agreement is found between theoretical and experimental results. The high accuracy of the B3LYP functional in reproducing experimental thermodynamic data for the present type of transition metal complexes is demonstrated by the fact that the differences between measured and computed thermodynamic parameters (DeltaG degree, DeltaH degrees , and -TDeltaS degree, in most cases are less than 2.0 kcal mol(-1). An attempt was made to investigate the kinetics of the formation of [Cu(II)(TPA(H))(O2-)]+ in THF and EtCN. Computed free energies of activation, DeltaG, are in good agreement with experimental results. However, an analysis of the partitioning of the free energy barriers in enthalpic and entropic contributions indicates that the computationally studied reaction pathway might differ from the one observed experimentally. PMID- 16471962 TI - Two three-dimensional metal-organic frameworks containing one-dimensional hydroxyl/carboxylate mixed bridged metal chains: syntheses, crystal structures, and magnetic properties. AB - Two novel three-dimensional complexes formulated as [M(3)(bime)2(mu3-OH)2(HO BDC)2]n (M = Co, 1; Cu, 2) [bime = 1,2-bis(imidazol-1'-yl)ethane, HO-H2BDC = 5 hydroxyisophthalic acid] have been hydrothermally synthesized and characterized. Both 1 and 2 exhibit similar structural frameworks resulting from one-dimensional metal/oxygen chains extended by HO-BDC, but the bridging modes of HO-BDC and coordination environments of metal centers are different. Complexes 1 and 2 crystallize in the orthorhombic system, space group Pbcn, a = 18.458(2) [18.2119(12) for 2] Angstroms, b = 12.0616(14) [11.6847(7)] Angstroms, c = 11.4859(14) [12.0688(6)] Angstroms, and Z = 4 (4). Magnetic studies show that 1 displays a slow magnetic relaxation, a large hysteresis, and distinct finite-size effects and 2 contains an antiferromagnetic chain. PMID- 16471961 TI - Solution and structural investigations of ligand preorganization in trivalent lanthanide complexes of bicyclic malonamides. AB - This report describes an investigation into the coordination chemistry of trivalent lanthanides in solution and the solid state with acyclic and preorganized bicyclic malonamide ligands. Two experimental investigations were performed: solution binding affinities were determined through single-phase spectrophotometric titrations and the extent of conformational change upon binding was investigated with single-crystal X-ray crystallography. Both experimental methods compare the bicyclic malonamide (BMA), which is designed to be preorganized for binding trivalent lanthanides, to an analogous acyclic malonamide. Results from the spectrophotometric titrations indicate that BMA exhibits a 10-100x increase in binding affinity to Ln(III) over acyclic malonamide. In addition, BMA forms compounds with high ligand-metal ratios, even when competing with water and nitrate ligands for binding sites. The crystal structures exhibit no significant differences in the nature of the binding between Ln(III) and the BMA or acyclic malonamide. These results support the conclusion that rational ligand design can lead to compounds that enhance the binding affinities within a ligand class. PMID- 16471963 TI - Syntheses and X-ray crystal structures of alpha- and beta-[XeO2F][SbF6], [XeO2F][AsF6], [FO2XeFXeO2F][AsF6], and [XeF5][SbF6)].XeOF4 and computational studies of the XeO2F+ and FO2XeFXeO2F+ cations and related species. AB - Reactions of XeO2F2 with the strong fluoride ion acceptors, AsF5 and SbF5, in anhydrous HF solvent give rise to alpha- and beta-[XeO2F][SbF6], [XeO2F][AsF6], and [FO2XeFXeO2F][AsF6]. The crystal structures of alpha-[XeO2F][SbF6] and [XeO2F][AsF6] consist of trigonal-pyramidal XeO2F+ cations, which are consistent with an AXY2E VSEPR arrangement, and distorted octahedral MF6- (M = As, Sb) anions. The beta-phase of [XeO2F][SbF6] is a tetramer in which the xenon atoms of four XeO2F+ cations and the antimony atoms of four SbF6- anions are positioned at alternate corners of a cube. The FO2XeFXeO2F+ cations of [FO(2)XeFXeO2F][AsF6] are comprised of two XeO2F units that are bridged by a fluorine atom, providing a bent Xe- - -F- - -Xe arrangement. The angle subtended by the bridging fluorine atom, a xenon atom, and the terminal fluorine atom of the XeO2F group is bent toward the valence electron lone-pair domain on xenon, so that each F- - -XeO2F moiety resembles the AX(2)Y(2)E arrangement and geometry of the parent XeO2F2 molecule. Reaction of XeF6 with [H3O][SbF6] in a 1:2 molar ratio in anhydrous HF predominantly yielded [XeF5][SbF6].XeOF4 as well as [XeO2F][Sb2F11]. The crystal structure of the former salt was also determined. The energy-minimized, gas-phase MP2 geometries for the XeO2F+ and FO2XeFXeO2F+ cations are compared with the experimental and calculated geometries of the related species IO2F, TeO2F-, XeO2(OTeF5)+, XeO2F2, and XeO2(OTeF5)2. The bonding in these species has been described by natural bond orbital and electron localization function analyses. The standard enthalpies and Gibbs free energies for reactions leading to XeO2F+ and FO2XeFXeO2F+ salts from MF5 (M = As, Sb) and XeO2F2 were obtained from Born Haber cycles and are mildly exothermic and positive, respectively. When the reactions are carried out in anhydrous HF at low temperatures, the salts are readily formed and crystallized from the reaction medium. With the exception of [XeO2F][AsF6], the XeO2F+ and FO2XeFXeO2F+ salts are kinetically stable toward dissociation to XeO2F2 and MF5 at room temperature. The salt, [XeO2F][AsF6], readily dissociates to [FO2XeFXeO2F][AsF6] and AsF5 under dynamic vacuum at 0 degree C. The decompositions of XeO2F+ salts to the corresponding XeF+ salts and O2 are exothermic and spontaneous but slow at room temperature. PMID- 16471964 TI - Bonding trends of thiosemicarbazones in mononuclear and dinuclear copper(I) complexes: syntheses, structures, and theoretical aspects. AB - Reactions of copper(I) halides with a series of thiosemicarbazone ligands (Htsc) in the presence of triphenylphosphine (Ph(3)P) in acetonitrile have yielded three types of complexes: (i) monomers, [CuX(eta1-S-Htsc)(Ph3P)2] [X, Htsc = I (1), Br (2), benzaldehyde thiosemicarbazone (Hbtsc); I (5), Br (6), Cl (7), pyridine-2 carbaldehyde thiosemicarbazone (Hpytsc)], (ii) halogen-bridged dimers, [Cu2(mu2 X)2(eta1-S-Htsc)2(Ph3P)2] [X, Htsc = Br (3), Hbtsc; I (8), furan-2-carbaldehyde thiosemicarbazone (Hftsc); I (11), thiophene-2-carbaldehyde thiosemicarbazone (Httsc)], and (iii) sulfur-bridged dimers, [Cu2X2(mu2-S-Htsc)2(Ph3P)2] [X, Htsc = Cl (4), Hbtsc; Br (9), Cl (10), pyrrole-2-carbaldehyde thiosemicarbazone (Hptsc); Br (12), Httsc]. All of these complexes have been characterized with the help of elemental analysis, IR, 1H, 13C, or 31P NMR spectroscopy, and X-ray crystallography (1-12). In all of the complexes, thiosemicarbazones are acting as neutral S-donor ligands in eta()S or mu2-S bonding modes. The Cu...Cu separations in the Cu(mu2-X)2Cu and Cu(mu2-S)2Cu cores lie in the ranges 2.981(1)-3.2247(6) and 2.813(1)-3.2329(8) Angstroms, respectively. The geometry around each Cu center in monomers and dimers may be treated as distorted tetrahedral. Ab initio density functional theory calculations on model monomeric and dimeric complexes of the simplest thiosemicarbazone [H2C=N-NH-C(S)-NH2, Htsc] have revealed that monomers and halogen-bridged dimers have similar stability and that sulfur bridged dimers are stable only when halogen atoms are engaged in hydrogen bonding with the solvent of crystallization or H2O molecules. PMID- 16471965 TI - Novel oxygen chirality induced by asymmetric coordination of an ether oxygen atom to a metal center in a series of sugar-pendant dipicolylamine copperII complexes. AB - Six sugar-pendant 2,2'-dipicolylamine (DPA) ligands (L1-3 and L'1-3) have been prepared. OH-protected and unprotected D-glucose, D-mannose, and D-xylose were attached to a DPA moiety via an O-glycoside linkage. X-ray crystallography of the copper(II) complexes (1-5) with these ligands revealed that the anomeric oxygen atom is coordinated to the metal center in the solid state, generating a chiral center at the oxygen atom. The CD spectra of these copper complexes in methanol or aqueous solution exhibit Cotton effects in the d-d transition region, which indicates that the ether oxygen atoms remain coordinated to the metal center and the oxygen-atom chirality is preserved even in solution. For complexes 1 and 2, the inverted oxygen-atom chirality and chelate-ring conformation in the solid state are well correlated with the mirror-image CD spectra in methanol solution. The concomitant inversion of the asymmetric configuration around the copper center was also observed in a methanol solution of complex 3 and a pyridine solution of complex 2. The square-pyramidal/octahedral copper(II) centers also exhibited characteristic absorption and CD spectra. PMID- 16471966 TI - De novo design of synthetic di-iron(I) complexes as structural models of the reduced form of iron-iron hydrogenase. AB - Simple synthetic di-iron dithiolate complexes provide good models of the composition of the active site of the iron-iron hydrogenase enzymes. However, the formally Fe(I)Fe(I) complexes synthesized to date fail to reproduce the precise orientation of the diatomic ligands about the iron centers that is observed in the molecular structure of the reduced form of the enzyme active site. This structural difference is often used to explain the fact that the synthetic di iron complexes are generally poor catalysts when compared to the enzyme. Herein, density functional theory computations are used for the rational design of synthetic complexes as structural models of the reduced form of the enzyme active site. These computations suggest several possible synthetic targets. The synthesis of complexes containing five-atom S-to-S linkers of the form S(CH2)2X(CH2)2S (X = CH2, NH, or O) or pendant functionalities attached to the three-carbon framework is one method. Another approach is the synthesis of asymmetrically substituted complexes, in which one iron center has strongly electron donating ligands and the adjacent iron center has strongly electron accepting ligands. The combination of a sterically demanding S-to-S linker and asymmetric substitution of the CO ligands is predicted to be a particularly effective synthetic target. PMID- 16471967 TI - Synthesis and characterization of silver(I) coordination networks bearing flexible thioethers: anion versus ligand dominated structures. AB - This report describes the synthesis and X-ray characterization of a series of L(n)AgX complexes wherein Ln = PhS(CH2)nSPh (n = 2, 4, 6, 10) and X = CF3SO3-, CF3COO-, CF3CF2COO-, CF3CF2CF2COO-, NO3-, and ClO4-. This study was undertaken in order to rationalize the structure of the coordination networks formed as a function of the anion coordinating strength and the ligand structure. The following complexes were examined: with L(2), CF3SO3- (1), CF3COO- (2), ClO4- (3); L4, CF3SO3- (4), CF3COO- (5), CF3CF2COO- (6), CF3CF2CF2COO- (7); L6, CF3COO .H2O (8), CF3CF2COO- (9), CF3CF2CF2COO- (10); and L10, NO3- (11). The anions selected are classified in three groups of increasing coordinating strength: perchlorates, fluorosulfonates, and perfluorocarboxylates. Except in two cases, all complexes form 2D-coordination networks. The 2D-network in 1 (L2, CF3SO3-) is made up of Ag(I) and L2, while the anion is only a terminal co-ligand that completes the trigonal coordination around Ag(I). In 4 (L4, CF3SO3-), a 1D coordination polymer, [Ag-L4-]infinity, is observed where the anions are coordinated to Ag(I) in a trigonal fashion. The perfluorocarboxylates form tetrameric units in a zigzag shape, but only with the L4 ligand. In these (6 and 7), the silver-silver distances are very short, especially those of the central bond, indicating the presence of weak Ag-Ag interactions. Dimers, with short silver-silver distances, are observed with ligands L2 and L6 and perfluorocarboxylates. In 8 (L6, CF3COO-.H2O), a 3D channel-like structure is built through water molecules that connect adjacent layers. An unusual stoichiometry is noted in 3 (L2, ClO4-, acetone); Ag:L is 4:2.5. In 11 (L10 and NO3-), the nitrate acts as a bidentate ligand and an [Ag-NO3-]infinity chain is formed. Adjacent chains are linked by the L10 ligands into a 2D-coordination network. PMID- 16471968 TI - Solvent tuning of the substitution behavior of a seven-coordinate iron(III) complex. AB - A detailed kinetic study of the substitution behavior of the seven-coordinate [Fe(dapsox)(L)2]ClO4 complex (H(2)dapsox = 2,6-diacetylpyridine bis(semioxamazide), L = solvent or its deprotonated form) with thiocyanate as a function of the thiocyanate concentration, temperature, and pressure was undertaken in protic (EtOH and acidified EtOH and MeOH) and aprotic (DMSO) organic solvents. The lability and substitution mechanism depend strongly on the selected solvent (i.e., on solvolytic and protolytic processes). In the case of alcoholic solutions, substitution of both solvent molecules by thiocyanate could be observed, whereas in DMSO only one substitution step occurred. For both substitution steps, [Fe(dapsox)(L)2]ClO4 shows similar mechanistic behavior in methanol and ethanol, which is best reflected by the values of the activation volumes (MeOH DeltaV(I) = +15.0 +/- 0.3 cm(3) mol(-1), DeltaV(II) = +12.0 +/- 0.2 cm(3) mol(-1); EtOH DeltaV(I) = +15.8 +/- 0.7 cm(3) mol(-1), DeltaV(II) = +11.1 +/- 0.5 cm(3) mol(-1)). On the basis of the reported activation parameters, a dissociative (D) mechanism for the first substitution step and a D or dissociative interchange (I(d)) mechanism for the second substitution step are suggested for the reaction in MeOH and EtOH. This is consistent with the predominant existence of alcoxo [Fe(dapsox)(ROH)(OR)] species in alcoholic solutions. In comparison, the activation parameters for the substitution of the aqua-hydroxo [Fe(dapsox)(H2O)(OH)] complex by thiocyanate at pH 5.1 in MES were determined to be DeltaH = 72 +/- 3 kJ mol(-1), DeltaS = +38 +/- 11 J K(-1) mol( 1), and DeltaV = -3.0 +/- 0.1 cm(3) mol(-1), and the operation of a dissociative interchange mechanism was suggested, taking the effect of pressure on the employed buffer into account. The addition of triflic acid to the alcoholic solutions ([HOTf] = 10(-3) and 10(-2) M to MeOH and EtOH, respectively) resulted in a drastic changeover in mechanism for the first substitution step, for which an associative interchange (Ia) mechanism is suggested, on the basis of the activation parameters obtained for both the forward and reverse reactions and the corresponding volume profile. The second substitution step remained to proceed through an I(d) or D mechanism (acidified MeOH DeltaV(II) = +9.2 +/- 0.2 cm(3) mol(-1); acidified EtOH DeltaV(II) = +10.2 +/- 0.2 cm(3) mol(-1)). The first substitution reaction in DMSO was found to be slowed by several orders of magnitude and to follow an associative interchange mechanism (DeltaS = -50 +/- 9 J K(-1) mol(-1), DeltaV(I) = -1.0 +/- 0.5 cm(3) mol(-1)), making DMSO a suitable solvent for monitoring substitution processes that are extremely fast in aqueous solution. PMID- 16471969 TI - Influence of ionic liquids bearing functional groups in dye-sensitized solar cells. AB - Ionic liquids containing the nitrile and vinyl functional groups attached to imidazolium cations combined with various anions, e.g., iodide, bis[(trifluoromethyl)sulfonyl]imide ([TFSI]-), or dicyanamide ([N(CN)2]-), have been prepared and characterized. These ionic liquids have been successfully used as electrolytes for dye-sensitized solar cells based on nanocrystalline TiO2 with the amphiphilic ruthenium sensitizer [ruthenium (4,4'-dicarboxylic acid-2,2' bipyridine)(4,4'-bis(p-hexyloxystyryl)-2,2'-bipyridine)][NCS]2 (coded K-19). The iodide salt was used in 3-methoxypropionitrile-based electrolytes, and the performances of both types of devices were evaluated on the basis of their photocurrent density-voltage characteristics and dark current measurements, demonstrating that the functional groups do not exert a detrimental effect on the performance. The solid-state structure of the nitrile-functionalized salt [C1C3CN(im)]I has also been established by single-crystal X-ray diffraction, revealing extensive hydrogen bonding between the cation protons and the iodide. PMID- 16471970 TI - GeO2 natrolite-type infinite four and eight R-containing layers in a 2D pure-Ge framework: Ge3O5(OH)4[C2N2H10]. AB - A new two-dimensional framework germanate, Ge3O5(OH)4[C2N2H10] (denoted ICMM-8), with a 3:9 Ge:O ratio has been synthesized, using a mixture of pyridine, water, and ethanol as the solvent and 1,4-diazabicyclo[2.2.2]octane and ethylenediamine as the structure-directing agents, under solvothermal conditions. The structure was determined by single-crystal X-ray diffraction. In this new compound, the GeO2 natrolite-type infinite four and eight R-containing layers appears for the first time in a pure GeO2 framework. The total 2D structure is built up from SBU 6, four tetrahedra, and two octahedra. The hydroxyl groups occupy four positions of each octahedral germanium atom. The compound is characterized by IR spectra and TGA-DTA. Crystal data: Ge3O5(OH)4[C2N2H10], monoclinic space group P2(1)/c; a = 11.3570(9) Angstroms, b = 8.8819(7) Angstroms, c = 9.9200(8) Angstroms, beta = 90.710(1), V = 1000.6(1) Angstroms(3), Z = 4, R(1) = 0.044 (I > 2(I)), and wR(2) = 0.1051 (all data). PMID- 16471971 TI - Transition state characterization for the reversible binding of dihydrogen to bis(2,2'-bipyridine)rhodium(I) from temperature- and pressure-dependent experimental and theoretical studies. AB - Thermodynamic and kinetic parameters for the oxidative addition of H2 to [Rh(I)(bpy)2]+ (bpy = 2,2'-bipyridine) to form [Rh(III)(H)2(bpy)2]+ were determined from either the UV-vis spectrum of equilibrium mixtures of [Rh(I)(bpy)2]+ and [Rh(III)(H)2(bpy)2]+ or from the observed rates of dihydride formation following visible-light irradiation of solutions containing [Rh(III)(H)2(bpy)2]+ as a function of H2 concentration, temperature, and pressure in acetone and methanol. The activation enthalpy and entropy in methanol are 10.0 kcal mol(-1) and -18 cal mol(-1) K(-1), respectively. The reaction enthalpy and entropy are -10.3 kcal mol(-1) and -19 cal mol(-1) K(-1), respectively. Similar values were obtained in acetone. Surprisingly, the volumes of activation for dihydride formation (-15 and -16 cm(3) mol(-1) in methanol and acetone, respectively) are very close to the overall reaction volumes (-15 cm(3) mol(-1) in both solvents). Thus, the volumes of activation for the reverse reaction, elimination of dihydrogen from the dihydrido complex, are approximately zero. B3LYP hybrid DFT calculations of the transition-state complex in methanol and similar MP2 calculations in the gas phase suggest that the dihydrogen has a short H-H bond (0.823 and 0.810 Angstroms, respectively) and forms only a weak Rh-H bond (1.866 and 1.915 Angstroms, respectively). Equal partial molar volumes of the dihydrogenrhodium(I) transition state and dihydridorhodium(III) can account for the experimental volume profile found for the overall process. PMID- 16471972 TI - Tl(I), Fe(II), and Co(II) complexes supported by a monoanionic N,N,N' heteroscorpionate ligand bis(3,5-di-tertbutylpyrazol-1-yl)-1-CH2NAr (Ar = 2,6 iPr2C6H3). AB - The lithium salt (L)Li(THF) (L- = bis(3,5-di-tertbutylpyrazol-1-yl)-1-CH2NAr, Ar = 2,6-iPr2C6H3) can be readily prepared from lithium bis(3,5-di-tertbutylpyrazol 1-yl)methide and the N-methyleneaniline H2C=NAr. This N,N,N'-heteroscorpionate lithium reagent can be transmetalated with Tl(OTf), FeCl2(THF)(1.5), and CoCl2 to yield the (L)Tl, (L)FeCl, and (L)CoCl complexes, respectively. Single crystal structural data for compounds (L)Li(THF), (L)Tl, (L)FeCl, and (L)CoCl reveal in each case the hapticity of the sterically demanding, monoanionic L- ligand to be kappa3-N3. PMID- 16471973 TI - Synthesis, structural evolution, and electrical properties of the novel Mo12 cluster compounds K(1+x)Mo12S14 (x = 0, 1.1, 1.3, and 1.6) with a tunnel structure. AB - The new structural type (1) K(2.3)Mo12S14 was prepared by solid-state reaction at 1500 degrees C in a sealed molybdenum crucible. The compound crystallizes in the trigonal space group P1c, Z = 2, (1) a = 9.1720(7) Angstroms, c = 16.403(4) Angstroms. Its crystal structure was determined from single-crystal X-ray diffraction data and consists of interconnected Mo12S14 units that form an original and unprecedented three-dimensional framework in which large tunnels are occupied randomly by a part of the K+ ions. The remaining K+ ions are localized between two consecutive Mo(12)S(14) units along the c axis. By carrying out topotactic oxydo-reduction reactions at low temperature (<100 degrees C), we were able to remove or insert K+ ions in the channels and thus form isostructural phases K(1+x)Mo12S14 (0 < or = x < or = 1.6). Thus, we have solved the crystal structures for the following three compositions: (2) K(2.1)Mo12S14, (3) KMo12S14, and (4) K(2.6)Mo12S14 ((2) a = 9.1476(4) A, c = 16.421(1) Angstroms; (3) a = 9.0797(9) Angstroms, c = 16.412(6) Angstroms; and (4) a = 9.1990(4) Angstroms, c = 16.426(4) Angstroms). Electrical resistivity measurements carried out on single crystals of K(2.3)Mo12S14 and KMo12S14 indicate that the former is semiconducting, whereas the latter is metallic. The evolution of the Mo-Mo distances with respect to the stoichiometry in potassium is discussed. PMID- 16471974 TI - Anion influence on the structures of a series of copper(II) metal-organic frameworks. AB - The main aim of the work herein presented is to investigate the influence of different anions on the overall structure of a series of metal-organic frameworks. The reactions between CuCl2, Cu(OAc)2, and CuSO4 and the two bipyridylurea ligands L1 and L2 [L1 = 1,3-bis(pyridin-4-ylmethyl)urea; L2 = 1,3 bis(pyridin-3-ylmethyl)urea; see Scheme 1 in paper] have been carried out and the crystal structure of five of the resulting metal-organic assemblies determined. These crystal structures have shown that the geometry and size of the corresponding anions together with their coordinating and hydrogen-bonding properties are essential in determining the final structures of the assemblies. Particularly interesting, because of their potential as nanoporous materials, are the assemblies obtained from the reaction between each of the two ligands (L1 and L2) and CuCl2, which yield noninterpenetrating 2D metal-organic layers made of squares of ca. 15 x 15 Angstroms. These layers stack on top of each other, producing infinite 3D channels filled with solvent molecules. The thermal stabilities of the five metal-organic frameworks prepared have been studied by means of thermogravimetric analysis. Preliminary X-ray powder diffraction studies of one of these metal-organic frameworks indicate that upon heating the assembly changes to a different crystalline structure. Interestingly, the original structure reforms upon exposure of this sample to traces of water. PMID- 16471975 TI - Metal-organic frameworks from homometallic chains of nickel(II) and 1,4 cyclohexanedicarboxylate connectors: ferrimagnet-ferromagnet transformation. AB - The hydrothermal reactions of nickel(II) nitrate with a mixture of the geometric cis and trans isomers of 1,4-cyclohexanedicarboxylic acid (1,4-chdc or C6H10(COOH)2) and a base yield three structurally different complexes, [Ni3(mu3 OH)2(mu4-cis-1,4-chdc)2(H2O)4].2H2O (1), [Ni(3)(mu3-OH)2(mu4-trans-1,4 chdc)2(H2O)4].4H2O (2), and [Ni(H2O)4(mu2-trans-1,4-chdc)], depending on the reaction conditions. The single-crystal X-ray structure analyses of 1 and 2 reveal segregation of the isomers and formation of frameworks based on infinite Ni3(OH)2(H2O)4 chains, acting as secondary building units, connected by either cis- or trans-chdc for 1 and 2, respectively. The frameworks sustain channels that house two or four water molecules, respectively, according to the size and shape of the channels that depend on the particular isomer. The structure of 3 consists of chains of square-planar Ni(H2O)4 bridged by trans-chdc. Magnetic data as a function of temperature and field of the virgin samples for 1 indicate long range ordering (LRO) to a ferrimagnetic ground state at 2.1 K that is reversibly transformed into a ferromagnet below 4.4 K upon partial dehydration and rehydration. Powder X-ray diffraction of 1, in its virgin state, after dehydration and after rehydration, confirms the stability of the framework. The magnetic data for 2 tend toward a LRO state to possibly a ferrimagnet below 2 K. The temperature dependence of the susceptibility of the two compounds is accounted for by the presence of both ferro- and antiferromagnetic exchanges within each chain via Ni-O-Ni and Ni-O-C-O-Ni pathways and weak coupling between neighboring chains via the 1,4-chdc unit. 3 is a uniform s = 1 antiferromagnetic chain (J/kB = 2.27(1) K). PMID- 16471977 TI - Structure of organic and metal-organic networks based on a bifunctional m terphenyl carboxylic acid. AB - A series of metal-organic frameworks (MOFs) based upon the ligand 2,6-diphenyl 1,4-dibenzoic acid [Ph2C6H2(CO2H)2]infinity have been prepared and characterized by X-ray crystallography. The networks exhibit a variety of topologies and coordination modes at the metal center. The reaction of the ligand with cobalt(II) nitrate or zinc(II) nitrate in methanol/pyridine results in the formation of isostructural 1-D chains [(Ph2C6H2(CO2)2)M(py)2(MeOH)]infinity, where M = Zn, Co; however, in the presence of ethanol and triethylamine, Zn(NO3)2 reacts to form a 2-D clay-like network, [(Ph2C6H2(CO2)2)Zn(EtOH)2]infinity. 2-D networks are also formed in similar reactions with copper(II) nitrate or silver(I) nitrate to give [(Ph2C6H2(CO2)(CO2H))2Cu(py)2]infinity, [(Ph2C6H2(CO2)CO2H))2Cu(py)4.2H2O](infinity), and [(Ph2C6H2(CO2)2)Ag2]infinity, respectively. The hydrogen-bonded chains formed by the ligand alone and with 4,4' dipyridyl are also described. PMID- 16471976 TI - Covalent and noncovalent interactions for [metal(dien)nucleobase](2+) complexes with L-tryptophan derivatives: formation of palladium-tryptophan species by nucleobase substitution under biologically relevant conditions. AB - The interaction of the complexes [Pd(dien)(1-MeCyt)]2+ (2) and [Pd(dien)(9 EtGH)]2+ (3) with the amino acids L-tryptophan (Trp) and N-acetyltryptophan (N AcTrp) was studied and compared with the previously studied platinum analogues [Pt(dien)(1-MeCyt)]2+ (4) and [Pt(dien)(9-EtGH)]2+ (5). Solid-state structures for 2 and 4 are reported. For the palladium complexes, the interaction is pH sensitive. Below pH 5, the noncovalent interaction with stacking between the aromatic amino acid residue and the metalated nucleobase was observed. Fluorescence quenching experiments indicated similar association constants for platinum and palladium derivatives 2-5. Unusual substitution of the model nucleobases 1-methylcytosine (1-MeCyt) and 9-ethylguanine (9-EtGH) by tryptophan was observed in the range of pH 5-11. The resulting species [Pd(dien)(Trp)]+ (6) and [Pd(dien)(N-AcTrp)]+ (7) were characterized using 1H NMR, 13C NMR, and ESI-MS spectroscopy with coordination indicated through the amino and deprotonated amido nitrogens, respectively. Complexes 6 and 7 were also obtained from a solution of [Pd(dien)Cl]+ (1) incubated with either Trp or N-AcTrp, respectively. PMID- 16471978 TI - Multisite effects on intervalence charge transfer in a clusterlike trinuclear assembly containing ruthenium and osmium. AB - The intervalence charge transfer (IVCT) properties of the mixed-valence forms of the diastereoisomers of the dinuclear [[Ru(bpy)2](mu-HAT)[M(bpy)2]]5+ (M = Ru or Os) complexes and the trinuclear heterochiral [[Ru(bpy)2]2[Os(bpy)2](mu-HAT)]n+ (n = 7, 8; HAT = 1,4,5,8,9,12-hexaazatriphenylene; bpy = 2,2'-bipyridine) species display a marked dependence on the nuclearity and extent of oxidation of the assemblies, while small differences are also observed for the diastereoisomers of the same complex in the dinuclear cases. The mixed-valence heterochiral [[Ru(bpy)2]2[Os(bpy)2](mu-HAT)]n+ (n = 7, 8) forms exhibit IVCT properties that are intermediate between those of the diastereoisomeric forms of the localized hetero-dinuclear complex [[Ru(bpy)2](mu-HAT)[Os(bpy)2]]5+ and the borderline localized-to-delocalized homo-trinuclear complex [[Ru(bpy)2]3(mu-HAT)]n+ (n = 7, 8). The near-infrared (NIR) spectrum of the +7 mixed-valence species exhibits both interconfigurational (IC) and IVCT transitions which are quantitatively similar to those in [[Ru(bpy)2](mu-HAT)[Os(bpy)2]]5+ and are indicative of the localized mixed-valence formulation [[Ru(II)(bpy)2]2[Os(III)(bpy)2](mu-HAT)]7+. The +8 state exhibits a new band attributable to an IVCT transition in the near infrared region. PMID- 16471979 TI - Synthesis, structure, and spectroscopic and magnetic properties of mesomorphic octakis(hexylthio)-substituted phthalocyanine rare-earth metal sandwich complexes. AB - The syntheses of new bis[octakis(hexylthio)phthalocyaninato] rare-earth metal(III) double-decker complexes [(C6S)8-Pc]2M (M = Gd(III), Dy(III), and Sm(III)) (2-4, respectively) are described. These compounds are very soluble in most common organic solvents. They have been fully characterized using elemental analysis, infrared, UV-vis spectroscopy, and mass spectrometry. The crystal structures of compounds 2-4 have been determined by X-ray diffraction on a single crystal. They are isostructural and crystallize in the monoclinic space group (space group C2/c). Their lattice constants have been determined in the following order: (2) a = 31.629(4) Angstroms, b = 32.861(4) Angstroms, c = 20.482(2) Angstroms, beta = 126.922(2) degrees, V = 17019(3) Angstroms(3); (3) a = 31.595(2) Angstroms, b = 32.816(2) Angstroms, c = 20.481(1) Angstroms, beta = 127.005(1) degrees, V = 16958(2) Angstroms(3); (4) a = 31.563(2) Angstroms, b = 32.796(2) Angstroms, c = 20.481(1) Angstroms, beta = 127.032 degrees, V = 16924(2) Angstroms(3). The magnetic properties of compounds 2-4 were studied, and it was revealed that the lanthanide ions and the radical delocalized on the two phthalocyanine rings are weakly interacting. The mesogenic properties of these new materials were studied by differential scanning calorimetry and optical microscopy. These phthalocyanine derivatives form columnar-hexagonal (Col(h)) mesophases. Thin films of bis[octakis(hexylthio)phthalocyaninato] rare-earth metal(III) double-decker complexes (2-4) were prepared by a spin-coating technique. Thermally induced molecular reorganization within films of bis[octakis(hexylthio)phthalocyaninato] rare-earth metal(III) double-decker complexes (2-4) was studied by the methods of ellipsometry, UV-vis absorption spectroscopy, and atomic force microscopy. Heat treatment produces molecular ordering, which is believed to be due to stacking interaction between neighboring phthalocyanine moieties. PMID- 16471980 TI - Cyclic voltammetry of Th(IV) in the room-temperature ionic liquid [Me3NnBu][N(SO2CF3)2]. AB - A Th(IV) compound, [Th(TFSI)4(HTFSI)].2H2O [where TFSI = N(SO2CF3)2], has been synthesized and characterized using elemental analysis, thermogravimetric analysis, and vibrational spectroscopy. The analysis suggests that the TFSI anion coordinates to the metal center via the sulfonyl oxygens as well as provides evidence for the coordination of HTFSI. The voltammetric behavior of this compound has been studied in the room-temperature ionic liquid [Me3NnBu][TFSI], and results show that Th(IV) is reduced to Th(0) in this ionic liquid in a single reduction step. Analysis of cyclic voltammograms shows that an insoluble product is being formed at the electrode surface, which is attributed to the formation of ThO2 by reaction with water. The E0 value for the reduction of Th(IV) to Th(0) has been determined to be -2.20 V (vs Fc+/Fc; -1.80 V vs SHE). A comparison of this E0 value with those obtained for Th(IV) reduction in a LiCl-KCl eutectic (400 degrees C), water, and nonaqueous solvents shows that the reduction in [Me3NnBu][TFSI] is easier to accomplish than that in these other solvents. PMID- 16471981 TI - Synthesis and reactivity of N-heterocycle-B(C6F5)3 complexes. 4. Competition between pyridine- and pyrrole-type substrates toward B(C6F5)3: structure and dynamics of 7-B(C6F5)3-7-azaindole and [7-Azaindolium]+[HOB(C6F5)3]-. AB - Reaction between 7-azaindole and B(C6F5)3 quantitatively yields 7-(C6F5)3B-7 azaindole (4), in which B(C6F5)3 coordinates to the pyridine nitrogen of 7 azaindole, leaving the pyrrole ring unreacted even in the presence of a second equivalent of B(C6F5)3. Reaction of 7-azaindole with H2O-B(C6F5)3 initially produces [7-azaindolium]+[HOB(C6F5)3]- (5) which slowly converts to 4 releasing a H2O molecule. Pyridine removes the borane from the known complexes (C6F5)3B pyrrole (1) and (C6F5)3B-indole (2), with formation of free pyrrole or indole, giving the more stable adduct (C6F5)3B-pyridine (3). The competition between pyridine and 7-azaindole for the coordination with B(C6F5)3 again yields 3. The molecular structures of compounds 4 and 5 have been determined both in the solid state and in solution and compared to the structures of other (C6F5)3B-N heterocycle complexes. Two dynamic processes have been found in compound 4. Their activation parameters (DeltaH = 66 (3) kJ/mol, DeltaS = -18 (10) J/mol K and DeltaH = 76 (5) kJ/mol, DeltaS = -5 (18) J/mol K) are comparable with those of other (C6F5)3B-based adducts. The nature of the intramolecular interactions that result in such energetic barriers is discussed. PMID- 16471982 TI - Nanoscale zinc antimonides: synthesis and phase stability. AB - Highly crystalline single-phase nanoparticles of the important thermoelectric materials Zn4Sb3 and ZnSb were prepared from solvochemically activated powders of elemental zinc and elemental antimony. Low-temperature reactions with reaction temperatures of 275-300 degrees C were applied using an excess of elemental zinc. The nanoscale thermoelectrics obtained were characterized by X-ray powder diffraction, transmission electron microscopy, and thermal analysis. nc-Zn4Sb3 showed particle sizes of 50-70 nm, whereas particle sizes of 15-20 nm were observed for nc-ZnSb. Calorimetric investigations showed an increased heat capacity, Cp, for nc-Zn4Sb3 with respect to the bulk material which could be reduced to the bulk value by annealing nc-Zn4Sb3 at 190 degrees C. Interestingly, nc-Zn4Sb3 showed exothermic decomposition into zinc-poorer ZnSn at 196 degrees C in an open system, indicating that Zn4Sb3 is metastable in nanocrystalline form at room temperature. PMID- 16471983 TI - Structural and electronic differences of copper(I) complexes with tris(pyrazolyl)methane and hydrotris(pyrazolyl)borate ligands. AB - Copper(I) complexes with tripodal nitrogen-containing neutral ligands such as tris(3,5-diisopropyl-1-pyrazolyl)methane (L1') and tris(3-tertiary-butyl-5 isopropyl-1-pyrazolyl)methane (L3'), and with corresponding anionic ligands such as hydrotris(3,5-diisopropyl-1-pyrazolyl)borate (L1-) and hydrotris(3-tertiary butyl-5-isopropyl-1-pyrazolyl)borate (L3-) were synthesized and structurally characterized. Copper(I) complexes [Cu(L1')Cl] (1), [Cu(L1')(OClO3)] (2), [Cu(L1')(NCMe)](PF6) (3a), [Cu(L1')(NCMe)](ClO4) (3b), [Cu(L1')(CO)](PF6) (4a), and [Cu(L1')(CO)](ClO4) (4b) were prepared using the ligand L1'. Copper(I) complexes [Cu(L3')Cl] (5) and [Cu(L3')(NCMe)](PF6) (6) with the ligand L3' were also synthesized. Copper(I) complexes [Cu(L1)(NCMe)] (7) and [Cu(L1)(CO)] (8) were prepared using the anionic ligand L1-. Finally, copper(I) complexes with anionic ligand L3- and acetonitrile (9) and carbon monoxide (10) were synthesized. The complexes obtained were fully characterized by IR, far-IR, 1H NMR, and 13C NMR spectroscopy. The structures of both ligands, L1' and L3', and of complexes 1, 2, 3a, 3b, 4a, 4b, 5, 6, 7, and 10 were determined by X-ray crystallography. The effects of the differences in (a) the fourth ligand and the counteranion, (b) the steric hindrance at the third position of the pyrazolyl rings, and most importantly, (c) the charge of the N3 type ligands, on the structures, spectroscopic properties, and reactivities of the copper(I) complexes are discussed. The observed differences in the reactivities toward O2 of the copper(I) acetonitrile complexes are traced back to differences in the oxidation potentials determined by cyclic voltammetry. A special focus is set on the carbonyl complexes, where the 13C NMR and vibrational data are presented. Density functional theory (DFT) calculations are used to shed light on the differences in CO bonding in the compounds with neutral and anionic N3 ligands. In correlation with the vibrational and electrochemical data of these complexes, it is demonstrated that the C-O stretching vibration is a sensitive probe for the "electron richness" of copper(I) in these compounds. PMID- 16471985 TI - Axial chirality and supramolecular interactions: the case of [(kappa2-P,P [PPh2NMe]2CO)Cu(I)Cl]2. AB - The title compound [(kappa2-P,P-[PPh(2)NMe}2CO)Cu(I)Cl]2 (4) is readily formed by the reaction between (PPh2NMe)2CO and copper(I) chloride. Compound 4 forms a two dimensional supramolecular network of Cu2Cl2-centered dimers that are linked by pi-pi interaction between the phenyl substituents on phosphorus in the solid state. Because of the nature of these pi-pi interactions, only one of the three possible enantiomers, the meso form, can be observed in the crystals. PMID- 16471984 TI - Utilization of the highly environment-sensitive emission properties of rhenium(I) amidodipyridoquinoxaline biotin complexes in the development of biological probes. AB - We report the synthesis and characterization of luminescent rhenium(I) amidodipyridoquinoxaline biotin complexes [Re(CO)3(dpqa)(L)](PF6) (dpqa = 2-(n butylamido)dipyrido[3,2-f:2',3'-h]quinoxaline; L = 4-(biotinamidomethyl)pyridine (py-4-CH2-NH-biotin) (1), 3-(N-((2-biotinamido)ethyl)amido)pyridine (py-3-CO-NH en-NH-biotin) (2), 4-(N-((6-biotinamido)hexanoyl)aminomethyl)pyridine (py-4-CH2 NH-cap-NH-biotin) (3)), and their biotin-free counterpart [Re(CO)3(dpqa)(py)](PF6) (py = pyridine (4)). Upon irradiation, these complexes exhibited intense triplet metal-to-ligand charge-transfer (3MLCT) (dpi(Re) --> pi(dpqa)) emission in fluid solutions at 298 K and in alcohol glass at 77 K. However, the emission became much weaker in aqueous buffer, probably due to the interactions of water molecules with the amide substituent of the dpqa ligand. These properties render the complexes good candidates as luminescent probes for hydrophobic media, such as the substrate-binding sites of proteins. The avidin binding properties of the new biotin complexes have been studied by 4' hydroxyazobenzene-2-carboxylic acid (HABA) assays, emission titrations, and competitive association and dissociation assays. Most importantly, the complexes showed a profound increase in emission intensities upon binding to avidin. Additionally, we found that the fluorescence of anthracene was quenched by these rhenium(I) complexes, and the 3MLCT emission of the complexes was also quenched by anthracene. On the basis of these findings, new homogeneous assays for biotin using these complexes, avidin, and anthracene-labeled avidin have been designed. PMID- 16471986 TI - Ruthenium(II) bipyridyl complexes as photolabile caging groups for amines. AB - The synthesis and characterization of a series of ruthenium bis(bipyridine) complexes where the inorganic moiety acts as a photolabile protecting group is described. Complexes of the type [Ru(bpy)2L2]+ where bpy = 2,2'-bipyridine and L = butylamine, gamma-aminobutyric acid, tyramine, tryptamine, and serotonin were studied by nuclear magnetic resonance, cyclic voltammetry, and electronic absorption spectroscopy. In all cases, ligands are coordinated by the amine group. The complexes are stable in water for several days and deliver one molecule of ligand upon irradiation with visible light (450 nm). These properties make them suitable for their use as biological caged compounds. PMID- 16471988 TI - Modified dipicolinic acid ligands for sensitization of europium(III) luminescence. AB - The effect of substitution at the 4 and 3,5 positions in the pyridine ring of europium(III) pyridine-2,6-dicarboxylate complexes has been investigated with particular emphasis on sensitization of the Eu3+ ion. Sensitization of the Eu3+ 615-nm emission was achieved through excitation of the ligands in which the 4 substituent was -H, -OH, and -Cl and the 3,5 position was -H. In these cases, the ligand-to-Eu3+ ratio was confirmed as being 3:1. The sensitization was found to increase following substitution of the 4 position in the order Cl > H > OH. This is attributed to energy transfer occurring from the ligands into different Eu3+ intra-atomic energy levels, with spin selection rules governing the efficiency of this process. The Eu3+ luminescence lifetime was measured and found to vary from 1.16 to 2.90 ms depending on the excitation energy, ligand, and solvent. For the case of the 3,5-dibromo-4-hydroxy derivative, no sensitization was observed and a ligand-to-Eu3+ ratio of 1:1 was found. The solubility of these complexes in water and their long emission lifetime make them attractive for use as probes in biological systems. PMID- 16471987 TI - Structure and bonding in monomeric iron(III) complexes with terminal oxo and hydroxo ligands. AB - We report on the structure and bonding in the title iron(III) complexes, containing the tris[(N'-tert-butylureayl)-N-ethyl]amine ligand, with density functional theory techniques. In agreement with the experimental data, a high spin electronic state is favored for all of the systems we considered. H bonds between the terminal oxo and hydroxo ligands and NH groups present in the organic ligand coordinated to the metal have a remarkable effect on the overall coordination geometry. In fact, the structure of model complexes without H bonds shows shorter Fe-O bond lengths. This is a consequence of the ability of the H bonds to stabilize a remarkable amount of electron density localized on the terminal oxo and hydroxo ligands. Energy analysis indicates that each H bond stabilizes the nonheme complexes by roughly 35 kJ/mol. Molecular orbital analysis indicates a reduction of two Fe-O bonding electrons on going from a complex with a terminal oxo ligand to a complex with a terminal hydroxo ligand. This reduction in the number of bonding electrons is also supported by frequency analysis. PMID- 16471989 TI - Di- and tetranuclear metal complexes with phenoxo bridges: synthesis, structures, and photoluminescent and electroluminescent properties. AB - Dinuclear and tetranuclear copper 2,6-bis(2-hydroxyphenyl)pyridine (H(2)L) complexes Cu2(L)2(py)2 (1) and Cu4(L)4(DMF) (2) were synthesized. The complexes 1 and 2 were characterized by elemental analyses, mass spectrometry, and single crystal X-ray diffraction analyses. 1 crystallizes in the monoclinic space group P2(1)/n with a = 13.330(2) Angstroms, b = 9.361(1) Angstroms, c = 14.676(1) Angstroms, beta = 100.94(1) degrees, V = 1798.1(3) Angstroms(3), and Z = 2. 2 crystallizes in the monoclinic space group P2(1)/n with a = 13.360(1) Angstroms, b = 14.884(1) Angstroms, c = 15.462(2) Angstroms, beta = 97.50(4) degrees, V = 3048.4(1) Angstroms(3), and Z = 2. Tetranuclear zinc complex Zn4(L)4(py)4 (3) was prepared and characterized by X-ray diffraction. 3 crystallizes in the triclinic space group P with a = 13.770(1) Angstroms, b = 15.465(1) A, c = 16.409(2) Angstroms, alpha = 88.877(9) degrees, beta = 88.035(4) degrees, gamma = 82.956(3) degrees, V = 3465.6(5) Angstroms(3), and Z = 2. The di- and tetranuclear complexes 1-3contain phenoxo bridges. 1 is a dinuclear complex with two Cu(II) centers, two py ligands, and two L ligands, and each L ligand donates its pyridyl ring and one of the phenolate groups to one metal and shares the other phenolate group between both metals, affording a Cu(2)(mu-O)(2) core. 2, in contrast, is a tetranuclear complex with four Cu(II) centers and four L ligands. Two of the L ligands have the same coordination mode as 1, and the other two L ligands donate their pyridyl rings to one metal and share both phenolate groups between four metals, resulting in three four-membered Cu2(mu-O)2 rings, which joined each other and showed great distortion from planarity. 3 is a tetranuclear complex with four Zn(II) centers, four pyridine ligands, and four L ligands, and the L ligands have the same coordination modes as those of 2. Single-crystal X-ray analysis showed that hydrogen-bonding and pi-pi stacking interactions exist in complexes 1 and 2 resulting in two- and three-dimensional molecular arrangements, and the parallel arrangement of the ligand in the crystal of complex 3 resulted in a close inter- and intramolecular pi-pi interactions. Investigation of the crystals, as well as an amorphous thin film and powder of 3, by photoluminescence (PL) allowed the effect of the molecular packing on the emission properties to be elucidated. Furthermore, the electroluminescent (EL) properties of 3 were examined by fabricating a multilayer device with structure of [ITO/NPB/(ZnL)(n)/Alq3/LiF/Al] (NPB = N,N'-bis(alpha-naphthyl)-N,N'-diphenyl (1,1'-biphenyl)-4,4'-diamine, Alq3 = tris(8-hydroxyquinolinato)aluminum). PMID- 16471990 TI - Volume-based thermodynamics: estimations for 2:2 salts. AB - The lattice energy of an ionic crystal, U(POT), can be expressed as a linear function of the inverse cube root of its formula unit volume (i.e., Vm(-1/3)); thus, U(POT) approximately 2I(alpha/Vm(1/3) + beta), where alpha and beta are fitted constants and I is the readily calculated ionic strength factor of the lattice. The standard entropy, S, is a linear function of Vm itself: S approximately kVm + c, with fitted constants k and c. The constants alpha and beta have previously been evaluated for salts with charge ratios of 1:1, 1:2, and 2:1 and for the general case q:p, while values of k and c applicable to ionic solids generally have earlier been reported. In this paper, we obtain alpha and beta, k and c, specifically for 2:2 salts (by studying the ionic oxides, sulfates, and carbonates), finding that U(POT)[MX 2:2]/(kJ mol(-1)) approximately 8(119/Vm(1/3) + 60) and S degree [MX 2:2]/(J K(-1) mol(-1)) approximately 1382V(m) + 16. PMID- 16471991 TI - [Au(CN)4]- as both an intramolecular and intermolecular bidentate ligand with [(tmeda)Cu(mu-OH)] dimers: from antiferro- to ferromagnetic coupling in polymorphs. AB - Two polymorphic products, [[Cu(tmeda)(mu-OH)}2Au(CN)4][Au(CN)4] (1) and [Cu(tmeda)(mu-OH)Au(CN)4]2 (2), were synthesized from {Cu(tmeda)(mu-OH)}(2)X(2) (tmeda = N,N,N',N'-tetramethylethylenediamine, X = ClO4-, BF4-) and 2 equiv of K[Au(CN)4], and their X-ray structures were determined. Both compounds have [Cu(tmeda)(mu-OH)}2(2+) dimers with [Au(CN)4]- units bound in the axial positions. However, in 1, two trans N-donor cyanides of each [Au(CN)4]- unit bind to adjacent copper(II) dimers, forming a 1-D chain, whereas complex 2 is molecular, with two mono-coordinated [Au(CN)4]- units. The 1-D polymorph 1 is formed from aqueous solution, while the molecular polymorph 2 is obtained with X = BF4- in methanol. The polymorphs have slightly different Cu-O-Cu angles, a key magnetostructural parameter, such that the 1-D chain 1, with an angle of 96.6(2) degrees, shows ferromagnetic interactions with 2J = +57.5 cm(-1) and g = 2.097, whereas the molecular complex 2, with an angle of 98.92(17) degrees, shows antiferromagnetic interactions with 2J = -143.6 cm(-1) and g = 2.047. A similar Cu(II) complex, [[Cu(tmeda)(mu-OH)]2Au(CN)4][ClO4].MeOH (3), was synthesized in methanol when X = ClO4-, in which the [Au(CN)4]- unit bridges the two Cu(II) centers within the dimer in an intramolecular fashion via cis N-donor cyanides. The average Cu-O-Cu angle of 98.4(2) degrees in 3 generates antiferromagnetic interactions with 2J = -64.8 cm(-1) and g = 2.214. Complexes 1-3 represent the first examples of [Cu(tmeda)(mu-OH)]2(2+) dimers with Cu-O-Cu angles under 100 degrees, thereby extending the range of 2J coupling constants for this moiety from 149 to 566 cm(-1). The switch to ferromagnetic interactions in 1 as a result of the coordinating, bridging [Au(CN)4]- anion suggests that cationic, dinuclear moieties that are typically antiferromagnetically coupled may, with an appropriate coordinating counterion, become ferromagnetic units. PMID- 16471992 TI - Synthesis and characterization of novel dinuclear copper(I) complexes. Dimerization of [CuL(PPh3)2] (L = methyl 3-hydroxy-3-(p-R-phenyl)-2 propenedithioate). AB - We report the synthesis of new copper(I) complexes 6a-e from methyl 3-hydroxy-3 (p-R-phenyl)-2-propenedithioate ligands. These complexes were characterized by IR and 1H, 13C, and 31P NMR spectroscopy. The expected O,S-coordination mode was confirmed by the X-ray diffraction studies of 6b and 6e. The unexpected dimerization of 6b-e leads to the formation of four novel dinuclear copper(I) compounds (7b-e). The dinuclear complex structure was fully established by the X ray diffraction analysis of 7a, in which the presence of a Cu-Cu interaction was observed. PMID- 16471993 TI - Magneto-structural correlations in Cu(tn)Cl2 (tn = 1,3-diaminopropane): two dimensional spatially anisotropic triangular magnet formed by hydrogen bonds. AB - A novel polymeric one-dimensional compound Cu(tn)Cl2 (tn = 1,3-diaminopropane) was prepared and structurally characterized, and its spectral, magnetic, thermodynamic, and thermal properties were studied. The unique structure shows ladderlike chains composed of Cu(II) atoms and chloro bridging ligands [Cu(-mu(3) Cl-)Cu2] running along the crystallographic c axis. The coordination geometry about copper (4 + 2) approximates that of a strongly elongated octahedron. The equatorial plane of the coordination octahedron is formed by a chelate N-bonded tn ligand and two chloro ligands. One of the chloro ligands is terminal, and the other one, mu3-Cl-, forms two additional longer bonds to the neighboring copper atoms and thus occupies the axial octahedral positions. The electronic ground state of the Cu(II) ion is of d(z)2 symmetry and suggests the activation of intraladder and interladder Cl...H-N hydrogen bonds as exchange paths that form a two-dimensional pattern of a triangular symmetry. The interaction due to the hydrogen bonds seems to play an important role in molecular packing and magnetic coupling. The studies of magneto-structural correlations including electron paramagnetic resonance measurements and thermodynamic and magnetic properties revealed a two-dimensional character of magnetic correlations with the effective intralayer exchange coupling J/k(B) approximately -3 K. No phase transition to the ordered state has been observed down to 60 mK. Cu(tn)Cl2 with the interlayer coupling J' approximately 10(-3)J and moderate intralayer interaction represents an excellent example of a two-dimensional magnetic system. PMID- 16471994 TI - Pentafluoronitrosulfane, SF5NO2. AB - The synthesis of pentafluoronitrosulfane, SF5NO2, is accomplished either by reacting N(SF5)3 with NO2 or by the photolysis of a SF5Br/NO2 mixture using diazo lamps. The product is purified by treatment with CsF and repeated trap-to-trap condensation. The solid compound melts at -78 degrees C, and the extrapolated boiling point is 9 degrees C. SF5NO2 is characterized by 19F, 15N NMR, IR, Raman, and UV spectroscopy as well as by mass spectrometry. The molecular structure of SF5NO2 is determined by gas electron diffraction. The molecule possesses C2v symmetry with the NO2 group staggering the equatorial S-F bonds and an extremely long 1.903(7) Angstroms S-N bond. Calculated bond enthalpies depend strongly on the computational method: 159 (MP2/6-311G++(3df)) and 87 kJ mol(-1) (B3LYP/6 311++G(3df)). The experimental geometry and vibrational spectrum are reproduced reasonably well by quantum chemical calculations. PMID- 16471995 TI - Coordination chemistry of Ga(C5Me4Ph): novel homoleptic d10 cluster complexes of palladium. AB - The synthesis and characterization of the novel sterically encumbered Ga(I) ligand GaCp(Ph) (1b) (Cp(Ph) = C5Me4Ph) is presented. GaCp(Ph) reacts with the Pd(0) source Pd2(dvds)3 (dvds = tetramethyldivinyldisiloxane) to give the trinuclear cluster [Pd3(GaCp(Ph))(mu2-GaCp(Ph))(mu3-GaCp(Ph))2(dvds)] (2a). 2a is the first example of a Ga(I)-containing cluster with a potentially labile olefinic ligand. It was found that 2a is an intermediate in the formation of the dinuclear cluster [Pd2(GaCp(Ph))2(mu2-GaCp(Ph))3] (2b), which is formed on reaction of 2a with GaCp(Ph). Both clusters were found to be labile toward GaCp, AlCp, or phosphines, giving substitution products in all cases. PMID- 16471996 TI - Aqueous spectroscopy and redox properties of carboxylate-bound titanium. AB - The aqueous chemistry of Ti(III) and Ti(IV) in two different chemical environments is investigated given its relevance to environmental, materials, and biological chemistry. Complexes of titanium with the carboxylate ligands citrate and oxalate, found ubiquitously in Nature, were synthesized. The redox properties were studied by using cyclic voltammetry. All the titanium citrate redox couples are quasi-reversible. Electrospray mass spectrometry of the Ti(III) citrate solution shows the presence of a 1:2 Ti/cit complex in solution, in contrast to the predominant 1:3 Ti/cit complex with Ti(IV). The change in the coordination of the ligand to the metal on reduction may explain the quasi-reversible behavior of the electrochemistry. The redox potentials for Ti(IV) citrate in water vary with pH. At pH 7, the approximate E(1/2) is less than -800 mV. This stated change in redox properties is considered in light of the previously reported Ti(IV) citrate solution speciation. Analogous speciation behavior is suggested from the EPR spectroscopy of Ti(III) citrate aqueous solutions. The g tensors are deduced for several pH-dependent species from the simulated data. The X-ray crystal structure of a Ti(III)(2) oxalate dimer Ti(2)(mu-C(2)O(4))(C(2)O(4))(2)(H(2)O)(6).2H(2)O (3), which crystallizes from water below pH 2, is reported. Complex 3 crystallizes in a monoclinic P2(1)/c space group with a = 9.5088(19) Angstroms, b = 6.2382(12) Angstroms, c = 13.494(3) Angstroms, V = 797.8(3) Angstroms(3), and Z = 2. The infrared spectroscopy, EPR spectroscopy, and cyclic voltammetry on complex 3 are reported. The cyclic voltammetry shows an irreversible redox couple approximately -196 mV which likely corresponds to the Ti(IV)(2)/Ti(III)Ti(IV) couple. The EPR spectroscopy on solid complex 3 shows a typical S = 1 triplet state spectrum. The solid follows non-Curie behavior, and the antiferromagnetic coupling between the two metal centers is determined to be -37.2 cm(-1). However, in solution the complex follows Curie behavior and supports a Ti(III)Ti(IV) oxidation state for the dimer. PMID- 16471997 TI - Platinum(II)-mediated coupling reactions of acetonitrile with the exocyclic nitrogen of 9-methyladenine and 1-methylcytosine. Synthesis, NMR characterization, and X-ray structures of new azametallacycle complexes. AB - The hydroxo complex cis-[L2Pt(mu-OH)]2(NO3)2, (L = PMePh2, 1a), in CH3CN solution, deprotonates the NH2 group of 9-methyladenine (9-MeAd) to give the cyclic trinuclear species cis-[L2Pt[9-MeAd(-H)]]3(NO3)3, (L = PMePh2, 2a), in which the nucleobase binds the metal centers through the N(1), N(6) atoms. In solution at room temperature, 2a slowly reacts with the solvent to form quantitatively the mononuclear azametallacycle cis-[L2PtNH=C(Me)[9-MeAd(-2H)]]NO3 (L = PMePh2, 3a), containing as anionic ligand the deprotonated form of molecule N-(9-methyl-1,9-dihydro-purin-6-ylidene)-acetamidine. In the same experimental conditions, the hydroxo complex with PPh3 (1b) forms immediately the insertion product 3b. Single-crystal X-ray analyses of 3a and 3b show the coordination of the platinum cation at the N(1) site of the purine moiety and to the N atom of the inserted acetonitrile, whereas the exocyclic amino nitrogen binds the carbon atom of the same CN group. The resulting six-membered ring is slightly distorted from planarity, with carbon-nitrogen bond distances for the inserted nitrile typical of a double bond [C(3)-N(2) = 1.292(7) Angstroms in 3a and 1.279(11) Angstroms in 3b], while the remaining CN bonds of the metallocycle are in the range of 1.335(8)-1.397(10) Angstroms. A detailed multinuclear 1H, 31P, 13C, and 15N NMR study shows that the nitrogen atom of the inserted acetonitrile molecule binds a proton suggesting for 3a,b an imino structure in solution. In DMSO and chlorinated solvents, 3a slowly releases the nitrile reforming the trinuclear species 2a, whereas 3b forms the mononuclear derivative cis-[L2Pt[9-MeAd(-H)]]NO3 (L = PPh3, 4b), in which the adeninate ion chelates the metal center through the N(6) and N(7) atoms. Complex 4b is quantitatively obtained when 1b reacts with 9 MeAd in DMSO and can be easily isolated if the reaction is carried out in CH(2)Cl(2). In CH(3)CN solution, at room temperature, 4b slowly converts into 3b indicating that the insertion of acetonitrile is a reversible process. A similar metal-mediated coupling reaction occurs when 1a,b react with 1-methylcytosine (1 MeCy) in CH(3)CN. The resulting complexes, cis-[L(2)PtNH=C(Me)[1-MeCy(-2H)]]NO3, (L = PMePh2, 5a and PPh3, 5b), contain the deprotonated form of the ligand N-(1 methyl-2-oxo-2,3-dihydro-1H-pyrimidin-4-ylidene)-acetamidine. The X-ray analysis of 5a shows the coordination of the metal at the N(3) site of the pyrimidine cycle and to the nitrogen atom of the acetonitrile, with features of the six membered metallocycle only slightly different from those found in 3a and 3b. In CD3CN/CH3(13)CN solution complexes 5a,b undergo exchange of the inserted nitrile, while in DMSO or chlorinated solvents they irreversibly release CH3CN to form species not yet fully characterized. No insertion of CH3CN occurs when the hydroxo complexes are stabilized by PMe3 and PMe2Ph. PMID- 16471998 TI - Dizinc alkoxides and amides supported by binucleating bis(amidoamine) ligands. AB - Several new dizinc complexes that are supported by dianionic bis(amidoamine) ligands are reported. Reaction of N,N'-bis(2-dimethylaminoethyl)dibenzofuran-4,6 diamine ((Me)LH(2)) with 2 equiv of EtZn(O(i)Pr) forms the dizinc bis(alkoxide) (Me)LZn2(O(i)Pr)2 (1), which was isolated in 76% yield. Similarly, (Me)LH2 reacts cleanly with EtZn(OPh) and EtZn(OCHPh2) to form (Me)LZn2(OPh)2 (2) and (Me)LZn2(OCHPh2)2 (3), respectively. The solid-state structures of 1 and 2 feature puckered [Zn2(mu-OR)2]2+ cores, with short intermetal separations (2.81 2.88 Angstroms). Overall, the molecules have approximate (noncrystallographic) C2v symmetry. The use of the more-hindered (i)Pr-substituted ligand N,N'-bis(2 diisopropylaminoethyl)dibenzofuran-4,6-diamine (i(Pr)LH2) to prepare zinc alkoxides gave similar results. Thus, reaction of i(Pr)LH2 with 2 equiv of EtZn(OPh), EtZn(OMe), EtZn(OCHPh2), and EtZn(OCH2Ph) forms i(Pr)LZn2(OPh)2 (4), i(Pr)LZn2(OMe)2 (5), i(Pr)LZn2(OCHPh2)2 (6), and i(Pr)LZn2(OCH2Ph)2 (7), respectively (isolated yields 48-63%). At 70 degrees C, C6D6 solutions of 6 undergo beta-hydride transfer with 2 equiv of benzaldehyde to form 7 and benzophenone in quantitative yield (according to 1H NMR spectroscopy). Benzene solutions of 1 react with 1 equiv of trimethylsilyl trifluoromethanesulfonate (Me3SiOTf) to form (Me)LZn2(O(i)Pr)(OTf) (8) in 70% isolated yield. In the solid state, 8 features a bridging alkoxide donor as well as a 1,3-bridging triflate group. The previously reported dinuclear organozinc species (Me)LZn2Ph2 (9) reacts with 1 equiv of tert-butylamine to form the protonolysis product (Me)LZn2(Ph)(NH(t)Bu) (10) in 66% isolated yield. The solid-state structure of 10 (two independent molecules) reveals a somewhat asymmetric [Zn2(mu-Ph)(mu NH(t)Bu)]2+ core with short Zn-Zn separations [2.6761(5) and 2.6518(5) Angstroms]. In CD2Cl2 solution, the Ph bridge of 10 undergoes rapid reversible cleavage. Cleavage of this bridging interaction followed by rotation about the Zn Ph bond and re-formation of the bridging interaction results in exchange of the inequivalent ortho (and meta) protons of the phenyl ligand. Variable-temperature 1H NMR spectroscopic data indicate that this exchange occurs with DeltaG = 12.7(1) kcal.mol(-1) (-27 degrees C). At 75 degrees C, toluene solutions of (Me)LH2 react with 2 equiv of EtZnNH(t)Bu to form the dizinc bis(amido) product (Me)LZn2(NH(t)Bu)2 (11) in 46% isolated yield. The solid-state structure of 11 (two independent molecules) features a puckered and fairly symmetric [Zn2(mu NH(t)Bu)2]2+ core with short intermetal separations [2.775(1), 2.760(1) Angstroms]. PMID- 16471999 TI - Synthesis, characterization, and hydrolysis of aluminum(III) compounds bearing the C6F5-substituted beta-diketiminate HC[(CMe)(NC6F5)]2 (L) ligand. AB - A series of Al(III) compounds containing the C6F5-substituted beta-diketiminate ligands LAlMeCl (2), LAlMe2 (3), LAlMeI (4), and LAlBr2 (5) (L = HC[(CMe)(NC6F5)]2) were synthesized and characterized. The hydrolysis of 2 and 4 in the presence of 1,3-diisopropyl-4,5-dimethylimidazol-2-ylidene as the hydrogen halide acceptor both lead to (LAlMe)2(mu-O) (6), a methylalumoxane derivative, which is the first hydrolysis product with the general formula of (RAlMe)(n)O. A comparison of the hydrolysis products of 2 and 4 with that of L'AlMeCl (L' = HC[(CMe)(NAr)]2, Ar = 2,6-iPr2C6H3) shows that with the C6F5-substituted beta diketiminate ligand, it was not possible to generate LAlMe(OH). This is obviously due to the stronger Bronsted acidity of the proton and the smaller size of the C6F5 group in this compound compared to that of the corresponding 2,6-iPr2C6H3 derivative. PMID- 16472000 TI - Solid-state 23Na and 7Li NMR investigations of sodium- and lithium-reduced mesoporous titanium oxides. AB - Mesoporous titanium oxide synthesized using a dodecylamine template was treated with 0.2, 0.6, and 1.0 equiv of Li- or Na-naphthalene. The composite materials were characterized by nitrogen adsorption, powder X-ray diffraction, X-ray photoelectron spectroscopy, elemental analysis, thermogravimetric analysis, and solid-state 23Na and 7Li NMR spectroscopy. In all cases the wormhole mesoporosity was retained as evidenced by BET surface areas from 400 to 700 m(2)/g, Horvath Kawazoe pore sizes in the 20 Angstroms range, and a lack of hysteresis in the nitrogen adsorption isotherms. Variable-temperature conductivity studies show that the Li-reduced materials are semiconductors, with conductivity values 3 orders of magnitude higher than those of the Na-reduced materials. Electrochemical measurements demonstrate reversible intercalation/deintercalation of Li+ ions into pristine mesoporous Ti oxides with good cycling capacity. Solid state 23Na NMR reveals two distinct Na environments: one corresponding to sodium ions in the mesoporous channels and the other corresponding to sodium ions intercalated into the metal framework. 23Na NMR spectra also indicate that the relative population of the framework site increases with increased reduction levels. Solid-state 7Li NMR spectra display a single broad resonance, which increases in breadth with increased reduction levels, though individual resonances inferring the presence of channel and framework Li species are not resolved. Comparisons of the lithium chemical shifts with published values suggests an "anatase-like structure" with no long-range order in the least reduced samples but a "lithium titanate-like structure" with no long-range order in the higher reduced materials. PMID- 16472001 TI - Pu(VI) hydrolysis: further evidence for a dimeric plutonyl hydroxide and contrasts with U(VI) chemistry. AB - A significant fraction of plutonium that is soluble in environmental waters and other aqueous solutions can be present as complexes of plutonyl, PuO2(2+). Few thermodynamic data are available for this ion, representing a problematic gap in plutonium chemistry and in the forecasting of radionuclide behavior under contamination and nuclear repository conditions. To address this need and more accurately determine the stoichiometry and stability of the basic hydrolytic products, we completed complimentary potentiometric and spectrophotometric studies of plutonium(VI) hydrolysis over the concentration range of 10(-2) to 10( 5) M Pu(VI). Dinuclear hydroxide species (PuO2)2(OH)2(2+) and (PuO2)2(OH)4(0)(aq) with hydrolysis constants log beta(2,2) = -7.79 +/- 0.20 and log beta(4,2) = 19.3 +/- 0.5 are indicated in all experiments of millimolar Pu(VI), 0.10 M NaNO3 solutions at 25 degrees C. At lower Pu(VI) concentrations, at and below 10(-4) M, the monomeric species PuO2OH+ and PuO2(OH)2(0)(aq) form with hydrolysis constants of log beta(1,1) = -5.76 +/- 0.07 and log beta(2,1) = -11.69 +/- 0.05, respectively. Distinct optical absorbance bands at 842 and 845 nm are reported for the mononuclear and dinuclear first hydrolysis species. Standard hydrolysis constants at zero ionic strength were calculated from the experimentally determined constants using the specific ion interaction theory. The Pu(VI) hydrolysis species and constants are compared with results from previous studies for plutonium and uranium. Major differences between uranyl and plutonyl hydrolysis are described. PMID- 16472002 TI - Formation and characterization of the uranium methylidene complexes CH2 = UHX (X = F, Cl, and Br). AB - The reactions between uranium atoms and CH3X (X = F, Cl, and Br) molecules are investigated in a solid argon matrix. The major products formed on ultraviolet irradiation are the CH2=UHX methylidene complexes. DFT calculations predict these triplet ground-state structures to be stable and to have significant agostic interactions. Parallels between the uranium and analogous thorium methylidene complexes are discussed. PMID- 16472003 TI - Syntheses, characterization, and X-ray crystal structures of beta-diketiminate group 13 hydrides, chlorides, and fluorides. AB - A series of organometallic compounds of group 13 metals supported by the sterically encumbered beta-diketiminate ligand containing hydrides, fluorides, chlorides, and bromide have been synthesized and structurally characterized. The synthetic strategy applied utilizes halide metathesis and reduction of metal chlorides to the corresponding hydrides. Thus, the reaction of LLi.OEt2 with MeMCl2 affords LM(Me)Cl (M = Al (1), Ga (2), In (3)) and LGaBr2 (4) with GaBr3. Reduction of LGa(Me)Cl with LiH.BEt3 leads to the formation of LGa(Me)H (10). Synthesis of LGaH(2) (12) has been accomplished by reacting LGaI2 (8) with LiH.BEt3. LAl(Me)Cl (1) and LAlH2 (6) have been converted to LAl(Me)F (5) and LAlF2 (7), respectively. The former was obtained in a reaction of LAl(Me)Cl with Me3SnF while the latter was isolated in a reaction of LAlH2 with BF3.OEt2. Similarly reaction of LGaI2 (8) with Me3SnF affords LGaF2 (9). Compounds reported herein have been characterized by elemental analyses, IR, NMR, EI-MS, and single crystal X-ray diffraction techniques. PMID- 16472004 TI - Photochemical and time resolved spectroscopic studies of intermediates relevant to iridium-catalyzed methanol carbonylation: photoinduced CO migratory insertion. AB - Photoreaction, time-resolved infrared (TRIR), and DFT studies were utilized to probe transformations between iridium complexes with possible relevance to the mechanisms of the iridium/iodide-catalyzed methanol carbonylation to acetic acid. Solution-phase continuous and laser flash photolysis of the tetraphenylarsonium salt of the fac-[CH3Ir(CO)2I3]- anion (1a) under excess carbon monoxide resulted in migratory insertion to give the acyl complex ion mer,trans [Ir(C(O)CH3)(CO)2I3]- (2a). The latter was isolated as its AsPh4+ salt, and its X ray crystal structure was determined. TRIR spectra indicate that several transients are generated upon flash photolysis of 1a. The principal photoreaction is CO dissociation, and this is proposed to generate the isomeric complexes fac [CH3Ir(CO)(Sol)I3]- (I(CO)(fac), Sol = solvent) and mer,trans-[CH3Ir(CO)(Sol)I3]- (I(CO)(mer)). I(CO)(fac) reacts with CO to regenerate 1a with a second-order rate constant (k(CO)) approximately 2.5 x 10(7) M(-1) s(-1) in ambient dichloroethane, while I(CO)(mer) is the apparent precursor to 2a. Kinetics studies indicate the photoinduced formation of a third intermediate (I(M)), hypothesized to be the anionic acyl complex fac-[Ir(C(O)CH3)(CO)(Sol)I3]-. In the absence of added CO, these intermediates undergo dimerization to form a mixture of isomers with the apparent formula [Ir(C(O)CH3)(CO)I3]2(2-). One of these dimers was isolated as the AsPh4+ salt, and the crystal structure was determined. Addition of excess pyridine to a solution of the dimers gave the neutral complex mer,trans [Ir(C(O)CH3)(CO)(py)2I2], which was characterized by FTIR, NMR, and X-ray crystallography. These transformations, especially the unprecedented photoinduced CO insertion reaction, are discussed and interpreted in terms of the factors favoring migratory insertion dynamics. PMID- 16472005 TI - Group 10 metal aminopyridinato complexes: synthesis, structure, and application as aryl-Cl activation and hydrosilane polymerization catalysts. AB - (4-Methyl-pyridin-2-yl)(trimethylsilanyl)amine (ApSi-H) and tert-butyl(4-methyl pyridin-2-yl)amine (AptBu-H) were synthesized via salt metathesis and aryl amination reactions, respectively. Lithiation of these two aminopyridines using n BuLi and the reactions with [(dme)NiCl2] (dme = dimethoxyethane) or [(cod)PdCl2] (cod = cyclooctadiene) in THF at low temperature gave rise--after workup in hexane--to group 10 amido compounds, [(ApSi)4Ni2], [(AptBu)2Pd], [(AptBu H)(AptBu)2Ni], [(AptBu)3(C2H5O)3Ni3OLi(thf)], and [(AptBu)2Ni(tBupy)2] (tBupy = 4 tert-butylpyridine). The aminopyridinato complexes were characterized by X-ray crystal structure analysis. The highly strained binding situation of the aminopyridinato ligands suggested that these compounds might be efficiently converted into catalytically active species. The applications of some of the synthesized complexes as Suzuki cross-coupling catalysts (activation of aryl chlorides) are described and [(ApSi)4Ni2] is a rare example of a "phosphine-free" catalyst system. A number of late transition metal complexes were found to successfully catalyze polymerization of MeH2SiSiH2Me toward soluble, linear poly(methylsilane). Remarkable activity was observed for [(ApSi)2Pd]. PMID- 16472007 TI - Epidemiology and pathophysiology of lower extremity peripheral arterial disease. AB - Peripheral arterial disease (PAD), a major cause of disability, loss of work, and lifestyle changes in the United States, is defined as obstruction of blood flow into an arterial tree excluding the intracranial or coronary circulations. PAD is mostly silent in its early stages, but when lesion obstruction exceeds 50%, it may cause intermittent claudication with ambulation. Further disease progression typically leads to rest pain or frank tissue loss. However, some patients may remain asymptomatic with severe disease because of extensive collateralization in the lower extremity. Estimates of the prevalence of intermittent claudication vary by population, from 0.6% to nearly 10%; the rate increases dramatically with age. Approximately 20% to 25% of patients will require revascularization, while fewer than 5% will progress to critical limb ischemia. Limb loss, although rare, is associated with severe disability and an overall poor prognosis, with 30% to 40% mortality in the first 24 months after limb loss. As with coronary artery disease, the most common cause of symptomatic obstruction in the peripheral arterial tree is atherosclerosis, a systemic inflammatory process in which cholesterol-laden plaque builds up in the artery and eventually blocks the lumen. Typical risk factors include age, gender, diabetes, tobacco abuse, hypertension, and hyperlipidemia. PMID- 16472008 TI - Diagnosis and risk assessment of lower extremity peripheral arterial disease. AB - Managing patients with peripheral arterial disease (PAD) requires an accurate assessment of the severity of the condition and the risk factors likely to predict disease progression. The spectrum of patient presentation ranges from asymptomatic to critical limb ischemia. Because about half of patients with PAD have coronary or cerebrovascular disease, the examination of presenting patients should be directed toward the entire cardiovascular system. The main diagnostic goal is to establish whether the symptoms are predominantly caused by PAD and to what degree the presenting problem is compounded by other comorbidities, such as diabetic neuropathy, arthritis, or venous disorders. The diagnostic process includes history taking, physical examination, noninvasive diagnostic testing, differential diagnosis, laboratory studies, and the use of the various imaging modalities, which in general are reserved for those PAD cases in which the clinician has already decided to intervene. PMID- 16472009 TI - Limitations of percutaneous transluminal angioplasty and stenting for the treatment of disease of the superficial femoral and popliteal arteries. AB - The long-term primary patency rates for percutaneous transluminal angioplasty (PTA) and stenting in treatment of disease of the superficial femoral and popliteal arteries have been disappointing. Three-year primary patency rates of approximately 50% have been reported for PTA and first-generation stents. Early results with newer nitinol stents appear promising; however, long-term data are lacking. One- and 2-year primary patency rates of 76% to 97% and 60% to 84%, respectively, have been reported with nitinol stents. An emerging concern with nitinol stents is the frequency of stent fracture (> or = 25%) after interventions for long-segment disease. A recent study has shown that stent fractures are associated with in-stent restenosis and with significantly lower primary patency at 1 year. The therapeutic success of PTA and stenting correlates with the complexity of the underlying disease. The best results are achieved when treating focal stenoses in patients with good distal runoff. Because endovascular interventions are preferred by patients, can be repeated, and preserve the option of bypass surgery, PTA and stenting are increasingly employed for diffuse and complex SFA disease. Clarifying the role of PTA and stenting in the treatment of complex SFA disease will require future studies that stratify results by both lesion anatomy and treatment indication. PMID- 16472010 TI - Contemporary endovascular treatment for disease of the superficial femoral and popliteal arteries: an integrated device-based strategy. AB - Endovascular therapy for disease of the superficial femoral artery (SFA) and the popliteal artery remains controversial. Percutaneous treatment of this arterial segment presents a particular technical challenge, as the extent of disease varies from short, focal, and stenotic to long, diffuse, and occluded lesions. Over the last 2 decades, multiple therapies have been evaluated, including simple balloon angioplasty, directional atherectomy, stenting (both balloon-expandable and self-expanding), and more recently, intra-arterial radiation, laser, and cryotherapy. Regardless of which modality is used, however, endovascular therapy as a revascularization strategy has the potential to improve symptoms and quality of life and, in selected patients, to avoid limb amputation. While percutaneous endovascular treatment has been historically associated with high procedural success and favorable short and intermediate-term patency rates, long-term clinical results have proven disappointing. Conventional balloon angioplasty is limited by elastic recoil, dissection, and restenosis. Balloon-expandable stents (particularly in the distal SFA) are associated with late stent deformation and mechanical compression, with resultant late clinical failure. Newer self expanding stents have shown improved initial results but have been limited by late mechanical fatigue and associated restenosis. With the development of several newer endovascular techniques in recent years, the possibilities for treating this condition have increased dramatically. Currently, no long-term comparative data exist regarding the role of these alternative technologies. This article summarizes and compares important data about new endovascular options for intervention therapy in SFA and popliteal disease. In addition, based on this analysis, we propose a contemporary treatment strategy, integrating older and newer technologies into a real-world algorithm. PMID- 16472011 TI - Infrainguinal lesion-specific device choices: round-table discussion. PMID- 16472012 TI - Endovascular management of infrainguinal disease. PMID- 16472013 TI - Review: the Rh blood group system: an historical calendar. PMID- 16472014 TI - Reactivity of FDA-approved anti-D reagents with partial D red blood cells. AB - Individuals whose RBCs are characterized as having a partial D phenotype may make anti-D if exposed to normal D+ RBCs; thus it is desirable that they be typed as D should they require blood transfusion or Rh immune globulin (RhIG) prophylaxis. Further, use of different anti-D reagents by blood centers and transfusion services can account for FDA-reportable errors. For this study, anti- D reagents for use in tube tests were obtained from three U.S. manufacturers. They included three examples of IgM monoclonal anti-D blended with monoclonal IgG anti-D, one IgM monoclonal anti-D blended with polyclonal IgG anti-D, and two reagents formulated with human anti-D in a high-protein diluent. One anti- D formulated for use by gel column technology was also tested. Direct agglutination tests by tube or gel were strongly positive (scores 9-12), with partial D RBCs of types DII, DIIIa, DIIIb, and DIVa. No reagent anti-D caused direct agglutination of DVI type 1, DVI type 2, or DFR phenotype RBCs. One tube anti-D reagent formulated with an IgM monoclonal anti-D plus a polyclonal IgG anti-D failed to cause direct agglutination of DVa, DBT, and R(0)(Har) RBCs, while DVa RBCs reacted weakly with two high-protein reagents formulated with human IgG anti-D. In contrast, the anti D used by gel column technology was strongly reactive (score 11) with DVa, DBT, and R(0)(Har) RBCs. The single monoclonal IgM-polyclonal IgG blended anti-D and the two high-protein reagents were also the only reagents that failed to react with R(0)(Har) RBCs by the IAT. Elimination of the test for weak D on all patient samples, using currently available FDA-licensed reagents, will ensure that partial D category VI (DVI) patients will type as D- for the purpose of RhIG prophylaxis and blood transfusion. However, RBCs of other partial D phenotypes will be classified as D+ in direct agglutination tests with some, if not all, currently available reagents. Testing donors for weak expression of D continues to be required, albeit that Rh alloimmunization by RBCs with a weak or partial D phenotype is uncommon. Further, because of differences in performance characteristics among FDA-approved reagents, conflicts between donor center D typing and transfusion service confirmatory test results are inevitable. PMID- 16472015 TI - Case report: immune anti-D stimulated by transfusion of fresh frozen plasma. AB - FFP has occasionally been reported to generate an immune response to RBC antigens (e.g., anti-D and anti-Fya). The Council of Europe requires that each unit of FFP have less than 6 x 10(9)/L RBCs. However, there is considerable variation internationally in the method of production and the level and assessment of RBC contamination of FFP. This study reports the case of a 63-year-old group B, D- man who received multiple transfusions of D- blood products over a 4-month period. Seven months later the patient's antibody screen remained negative and he was transfused with seven units of D- RBCs and six units of FFP, four of which were D+. Two months later anti-D, -E, and -K were detected in his plasma. Although the anti-E and anti-K could have resulted from transfusion of antigen positive RBCs, the anti-D could have resulted only from transfusion of the D+ FFP. The D status of FFP is currently not considered when selecting products for transfusion even though the D antigen is highly immunogenic and the level of RBC contamination of FFP is not always known. This case highlights that transfusion of FFP is a stimulus for RBC antibodies and that when a patient has had a recent transfusion of FFP, consideration should be given to obtaining a sample for pretransfusion testing within 3 days before a scheduled RBC transfusion. In addition, the D status of FFP should be considered before administering FFP to premenopausal D- women. PMID- 16472016 TI - Incidence of weak D in blood donors typed as D positive by the Olympus PK 7200. AB - The incidence of weak D has been reported to be between 0.23 and 0.5 percent in Europe and 3.0 percent in the United States. All studies were performed before the introduction of monoclonal anti-D reagents. Using current commercial reagents, this study evaluated D+ samples for the presence of weak D. D+ donors, typed by the Olympus PK 7200, using diluted monoclonal blend anti-D and diluted polyclonal anti-D, were selected by sampling batches of 100 to 200 samples from the previous day's collection. Anti-D reagents used on the Olympus PK 7200 are required to detect RBCs with the weak D phenotype which do not agglutinate at immediate spin (IS) when tested with polyclonal anti-D by manual tube methods. More than 95 percent of donors tested were Caucasian. Using tube tests with two different monoclonal blend anti-D reagents and one polyclonal anti-D typing reagent, the presence or absence of the D antigen was evaluated after the IS reading. Donors found negative or weakly positive (< 2+) at IS were further typed for weak D by the IAT. The weak D samples were RHD genotyped by allele-specific PCR. Of 1,005 donors tested, 4 (0.4%) were classified as weak D by one or more anti-D reagents. Polyclonal anti-D reagent demonstrated weaker reactions when compared with the monoclonal blends. All weak D samples were found positive for exon 4, intron 4, and exon 10, a finding consistent with most D+ samples. The incidence of weak D found in this study is not significantly different from that found in earlier studies using polyclonal anti-D reagents. PMID- 16472017 TI - Review: the Rh blood group D antigen... dominant, diverse, and difficult. PMID- 16472018 TI - Violence in rural communities: youth speak out! AB - INTRODUCTION: The current available literature does not present the viewpoints of rural youths regarding the meaning of violence. DESIGN: A mixed method exploratory, descriptive study was conducted to generate information from rural youth about violence. The qualitative phase of the exploratory, descriptive study is reported here. RESULTS: Interviews with fifty-two youth (20 males and 32 females), ranging in age from 11 to 19 years in grades 7-12 in the public school system were conducted in two rural communities. The participants initially defined violence as a physical act with intent to harm, but did not consider weapon carrying as an issue in their communities. They identified alcohol consumption as a trigger for violence and described planned fights that occurred away from school grounds. Physical fights were not limited to the male students. The youth openly stated that violence exists in rural communities and felt isolated from adults with regard to understanding the youths' experiences with violence. CONCLUSION: It is important to generate information about violence directly from rural youth. Programs to reduce violence that include the youth are important to pursue. PMID- 16472019 TI - Validation of S. pombe sequence assembly by microarray hybridization. AB - We describe a method to make physical maps of genomes using correlative hybridization patterns of probes to random pools of BACs. We derive thereby an estimated distance between probes, and then use this estimated distance to order probes. To test the method, we used BAC libraries from Schizzosaccharomyces pombe. We compared our data to the known sequence assembly, in order to assess accuracy. We demonstrate a small number of significant discrepancies between our method and the map derived by sequence assembly. Some of these discrepancies may arise because genome order within a population is not stable; imposing a linear order on a population may not be biologically meaningful. PMID- 16472020 TI - A grammatical theory for the conformational changes of simple helix bundles. AB - Polymers, including biomolecules such as proteins, have two particularly important types of single-molecule transitions: a helix-coil transition, driven by interactions that are local in the chain, and a collapse transition, driven by nonlocal interactions. A long-standing challenge of polymer statistical mechanics has been to deal with both types of transition in a single theoretical framework. The simplest paradigmatic problem would be a theory of helix-bundle folding. Here, we show how the machinery of formal grammars, originally developed in the context of linguistic analysis and programming-language compilation, provides a simple and general way to combine the Zimm-Bragg model of alpha-helices with the model of Chen and Dill for nonlocal interactions in antiparallel polymeric systems. We use a well-known construction in the theory of formal grammars to give the statistical mechanical partition function for two-helix bundles. Predictions are shown to be quite good in comparison to exact enumerations within a lattice model. PMID- 16472022 TI - Signal deconvolution based expression-detection and background adjustment for microarray data. AB - Background adjustment is an essential stage in analyzing DNA microarrays. Discriminating expressed genes from unexpressed ones (expression detection), and estimating the expression levels of weakly expressed genes, critically depend on accurate treatment of the background intensity. Current methods for background adjustment either do not deal with nonspecific hybridization or strongly depend on the reliability of control probes. Existing model-based methods have limited accuracy. A new platform-independent background adjustment algorithm is presented. The algorithm relies on the deconvoluted experimental signal distribution for evaluating the expression probability and adjusting the background of each probe. Considering expression detection, it is shown, for two channels cDNA arrays and for the Affymetrix GeneChip platform, that the algorithm performs at least as good or better than control-probes-based algorithms. For the Affymetrix GeneChip arrays, it is further shown that the algorithm outperforms the robust multiarray (RMA) expression measure in estimating genomewide expression levels. PMID- 16472021 TI - Flexible secondary structure based protein structure comparison applied to the detection of circular permutation. AB - We present a novel method for structural comparison of protein structures. The approach consists of two main phases: 1) an initial search phase where, starting from aligned pairs of secondary structure elements, the space of 3D transformations is searched for similarities and 2) a subsequent refinement phase where interim solutions are subjected to parallel, local, iterative dynamic programming in the areas of possible improvement. The proposed method combines dynamic programming for finding alignments but does not restrict solutions to be sequential. In addition, to deal with the problem of nonuniqueness of optimal similarities, we introduce a consensus scoring method in selecting the preferred similarity and provide a list of top-ranked solutions. The method, called FASE (flexible alignment of secondary structure elements), was tested on well-known data and various standard problems from the literature. The results show that FASE is able to find remote and weak similarities consistently using a reasonable run time. The method was tested (using the SCOP database) on its ability to discriminate interfold pairs from intrafold pairs at the level of the best existing methods. The method was then applied to the problem of finding circular permutations in proteins. PMID- 16472023 TI - Alignment with context dependent scoring function. AB - In the paper by Gambin et al. (2002) we introduced the model of contextual alignment of biological sequences. It is an extension of the classical alignment, in which the cost of a substitution depends on the surrounding symbols. Consequently, in this model the cost of transforming one sequence into another depends on the order of editing operations. In this paper, we strengthen some of our results which concern reconstructing (the representation of) all the orders of operations which yield this optimal cost. We also present a procedure to construct context-dependent substitution tables and discuss the distribution of scores of local contextual alignment, which is shown to follow the extreme value distribution in the gap-free, reduced context case. We also demonstrate a linear time algorithm to compute the optimal local and global alignment without gaps. PMID- 16472024 TI - Recognition and classification of histones using support vector machine. AB - Histones are DNA-binding proteins found in the chromatin of all eukaryotic cells. They are highly conserved and can be grouped into five major classes: H1/H5, H2A, H2B, H3, and H4. Two copies of H2A, H2B, H3, and H4 bind to about 160 base pairs of DNA forming the core of the nucleosome (the repeating structure of chromatin) and H1/H5 bind to its DNA linker sequence. Overall, histones have a high arginine/lysine content that is optimal for interaction with DNA. This sequence bias can make the classification of histones difficult using standard sequence similarity approaches. Therefore, in this paper, we applied support vector machine (SVM) to recognize and classify histones on the basis of their amino acid and dipeptide composition. On evaluation through a five-fold cross-validation, the SVM-based method was able to distinguish histones from nonhistones (nuclear proteins) with an accuracy around 98%. Similarly, we obtained an overall >95% accuracy in discriminating the five classes of histones through the application of 1-versus-rest (1-v-r) SVM. Finally, we have applied this SVM-based method to the detection of histones from whole proteomes and found a comparable sensitivity to that accomplished by hidden Markov motifs (HMM) profiles. PMID- 16472025 TI - A multiobjective evolutionary method for the design of peptidic mimotopes. AB - Peptides that mimic protein epitopes are interesting drug candidates. However, the design of effective peptidic drugs is difficult for several reasons, such as the fast degradation of peptides, their high flexibility, and thus high entropy loss on binding to the target. We therefore propose an in silico method for the automated design of peptides that are optimal with respect to several objectives. We present a Pareto-based multiobjective evolutionary algorithm for in silico peptide design. Using a simple molecular model, we apply the method to the design of peptides that (a) mimic antibody epitopes of the proteins thrombin and blood coagulation factor VIII, respectively, that (b) are short, and (c) are conformationally stable. PMID- 16472026 TI - Prediction of membrane proteins in Mycobacterium tuberculosis using a support vector machine algorithm. AB - We report our finding of linear clustering of signal sequences at the N-terminus of M.tb membrane proteins, directing membrane localization. Although it is widely accepted that membrane proteins have signal peptides at the N-terminus, statistical ensemble analysis of Support Vector Machine prediction results indicate that M.tb membrane proteins have embedded N-terminal sequence patterns beyond the signal peptides previously identified in E. coli. The additional patterns at the N-terminus of M.tb membrane proteins may have correlations to their unique enzymatic functions and unusual characteristics such as membrane interaction in pathogenes. PMID- 16472031 TI - Looking back, looking forward. PMID- 16472032 TI - Are we creating relevant behavioral medicine research? Show me the evidence! PMID- 16472033 TI - Practical behavioral trials to advance evidence-based behavioral medicine. AB - There is a well-documented gap between research and practice in many areas of behavioral medicine. This gap is due in part to limitations in the capacity of the research database to address questions that are of central concern to clinicians, administrative decision makers, and policymakers. Thus, there has been a call for "practical clinical trials" that compare clinically viable alternative interventions and assess multiple outcomes important for clinical and policy decisions in diverse patient populations and settings. Such trials offer great potential, and they raise interesting challenges regarding optimal research design and source of funding. We discuss issues related to practical clinical trials in behavioral medicine, propose a need for practical behavioral trials (PBTs), and describe design features that will facilitate clinical and policy decision making. This type of PBT can help to close the gap between research and practice and advance the field of evidence-based behavioral medicine. We discuss potential challenges and objections to PBTs and conclude by providing recommendations for the design, conduct, reporting, and review of practical trials. PMID- 16472034 TI - Fulfilling our assignment to improve the health of all: good science just isn't enough. PMID- 16472035 TI - Shared and unique contributions of anger, anxiety, and depression to coronary heart disease: a prospective study in the normative aging study. AB - BACKGROUND: Anger, anxiety, and depression have each been identified as risk factors for coronary heart disease (CHD). Whether the apparent risk is a function of unique aspects of each emotion or due to a shared underlying dimension of negative affectivity is unclear. PURPOSE: The goal of this study was to assess shared and unique contributions of anger, anxiety, and depression to incident CHD. METHODS: Data are from the Veterans Administration Normative Aging Study, an ongoing cohort of older men. Measures of anger, anxiety, and depression were obtained from 1,306 men completing the revised Minnesota Multiphasic Personality Inventory in 1986. From these measures we derived three near-orthogonal scales termed iso(lated)-anger, iso-anxiety, and iso-depression and a fourth scale measuring general distress. RESULTS: During an average of 10.9 years of follow up, 161 cases of incident CHD occurred. When considered individually, iso anxiety, iso-anger, and shared general distress were each associated with CHD risk. When all emotions were considered simultaneously, only iso-anxiety and shared general distress were associated with incident CHD. CONCLUSIONS: Considering shared versus unique aspects of negative emotions may clarify the nature of their apparent toxicity in relation to CHD risk. General distress shared across negative emotions is an important component in the emotion-CHD relation. Aspects of anxiety may also independently increase CHD risk. PMID- 16472036 TI - Frequency of attendance at religious services and leisure-time physical activity in American women and men: the Third National Health and Nutrition Examination Survey. AB - BACKGROUND: Studies are lacking from representative samples of total populations and Hispanic Americans on the association of religiousness and leisure time physical activity (LTPA). PURPOSE: The objective is to test the hypothesis that frequency of attendance at religious services is positively associated with LTPA. METHODS: The Third National Health and Nutrition Examination Survey (NHANES III) included 11,820 persons 20 years of age and older with complete data on self reported frequency of attendance at religious services, LTPA, and mobility limitation. RESULTS: Among older women with no mobility limitation, infrequent attenders had significantly higher prevalence of no LTPA (odds ratio = 1.4; 95% confidence interval = 1.1, 1.9; p = .02), but infrequent attenders were not significantly different from others in prevalence of moderate or vigorous LTPA 5 or more times per week after adjustment for sociodemographic variables and health status. No significant associations were seen at 20 to 59 years of age or in men. CONCLUSIONS: In older women without mobility limitation, less than weekly attendance at religious services was associated with greater prevalence of no LTPA even after controlling for health status. Significant adjusted associations were not seen in men or younger women. PMID- 16472037 TI - Individual, social environmental, and physical environmental influences on physical activity among black and white adults: a structural equation analysis. AB - BACKGROUND: Social ecological models suggest that conditions in the social and physical environment, in addition to individual factors, play important roles in health behavior change. Using structural equation modeling, this study tested a theoretically and empirically based explanatory model of physical activity to examine theorized direct and indirect effects of individual (e.g., motivation and self-efficacy), social environmental (e.g., social support), and physical environmental factors (e.g., neighborhood quality and availability of facilities). METHOD: A community-based sample of adults (N = 910) was recruited from 2 public health centers (67% female, 43% African American, 43% < $20,000/year, M age = 33 years) and completed a self-administered survey assessing their current physical activity level, intrinsic and extrinsic motivation for physical activity, perceived social support, self-efficacy, and perceptions of the physical environment. RESULTS: Results indicated that (a) perceptions of the physical environment had direct effects on physical activity, (b) both the social and physical environments had indirect effects on physical activity through motivation and self-efficacy, and (c) social support influenced physical activity indirectly through intrinsic and extrinsic motivation. For all forms of activity, self-efficacy was the strongest direct correlate of physical activity, and evidence of a positive dose-response relation emerged between self efficacy and intensity of physical activity. CONCLUSIONS: Findings from this research highlight the interactive role of individual and environmental influences on physical activity. PMID- 16472038 TI - A heuristics approach to understanding cancer risk perception: contributions from judgment and decision-making research. AB - BACKGROUND: The likelihood judgments that people make about their risks for cancer have important implications. At the individual level, risk estimates guide protective actions, such as cancer screening. However, at the extremes, exaggerated risk judgments can also lead to anxiety that degrades quality of life or to aggressive self-protective actions that are unwarranted given the objective risks. At the policy level, risk judgments may serve as an indicator of societal perceptions of the "war" against cancer. Using risk judgments, the public expresses its belief about whether we are winning. PURPOSE: We present theoretical perspectives from judgment and decision making, illustrate how they can explain some of the existing empirical findings in the cancer risk literature, and describe additional predictions that have not yet been tested. CONCLUSIONS: Overall, we suggest that theories from the judgment and decision making perspective offer a potentially powerful view for understanding and improving risk judgments for cancer and other diseases. PMID- 16472039 TI - Effects of a CHANGE intervention to increase exercise maintenance following cardiac events. AB - BACKGROUND: Despite participation in a cardiac rehabilitation program, there is a downward trajectory of exercise participation during the year following a cardiac event. PURPOSE: The purpose of this study was to test the effectiveness of CHANGE (Change Habits by Applying New Goals and Experiences), a lifestyle modification program designed to increase exercise maintenance in the year following a cardiac rehabilitation program. The CHANGE intervention consists of 5 small-group cognitive-behavioral change counseling sessions in which participants are taught self-efficacy enhancement, problem-solving skills, and relapse prevention strategies to address exercise maintenance problems. METHOD: Participants (N = 250) were randomly assigned to the CHANGE intervention (supplemental to usual care) or a usual-care-only group. Exercise was measured using portable wristwatch heart rate monitors worn during exercise for 1 year. Cox proportional hazards regression was used to determine differences in exercise over the study year between the study groups. RESULTS: Participants in the usual-care group were 76% more likely than those in the CHANGE group to stop exercising during the year following a cardiac rehabilitation program (hazard ratio = 1.76, 95% confidence interval = 1.08-2.86, p = .02) when adjusting for the significant covariates race, gender, comorbidity, muscle and joint pain, and baseline motivation. Most participants, however, had less than recommended levels of exercise amount and intensity. CONCLUSIONS: Counseling interventions that use contemporary behavior change strategies, such as the CHANGE intervention, can reduce the number of individuals who do not exercise following cardiac events. PMID- 16472040 TI - Interpersonal factors predict increased desire for hastened death in late-stage cancer patients. AB - BACKGROUND: Cancer patients at the end of life sometimes express a wish that death would come quickly, but this desire for hastened death (DHD) remains little understood. Relationships with spousal caregivers may play a role in patients' DHD. PURPOSE: This study examined factors that could predict an increase in the DHD in late-stage cancer patients over the course of 4 months, including marital and caregiving variables that have not previously been examined. METHOD: Patients completed the Schedule of Attitudes Toward Hastened Death and other measures, including the Dyadic Adjustment Scale. Caregivers were asked how many hours they spent weekly in caregiving activities and completed the Caregiver Demands Scale. Approximately 4 months later, DHD was reassessed in surviving patients. RESULTS: Sixty caregiver/patient dyads completed all measures. Desire for hastened death was generally low at both assessments; however, more depression and greater dyadic adjustment reported by patients, and more hours spent in caregiving activities by spouses, each independently predicted increased DHD in patients at the Time 2 assessment. CONCLUSIONS: Findings suggest that issues related to spousal caregivers play an important role in the course of DHD in cancer patients at the end of life. PMID- 16472041 TI - Information-Motivation-Behavioral Skills (IMB) Model: testing direct and mediated treatment effects on condom use among women in low-income housing. AB - BACKGROUND: The Information-Motivation-Behavioral Skills (IMB) model of HIV preventive behavior (1-4) specifies that treatment effects on behavior occur largely as the result of treatment effects on behavioral skills, which follow from effects on information and motivation. PURPOSE: The objective was to determine whether the variables specified by the IMB model of HIV preventive behavior (1-4) accounted for the relation between an IMB-based treatment and resulting HIV preventive behavior (condom use). METHOD: Women (n = 557) living in 18 low-income housing developments in 5 geographically dispersed cities were recruited to participate in an HIV-prevention study. Women (within housing developments) were randomly assigned to receive an IMB-based, HIV risk-avoidance intervention or a comparison intervention. Baseline and posttreatment (16 months after baseline) data were collected on condom use information, motivation (social norms, attitudes, intentions, and perceived risk), enactment of behavioral skills (condom negotiation and procurement), and rates of condom use in the past 2 months. RESULTS: The IMB intervention led to a 12% to 16% increase in condom use rates over the course, whereas the comparison intervention led to 2% decrease. In addition, the IMB treatment led to greater increases in condom use information, in the intentions and social norms components of motivation and the condom procurement and condom conversations components of behavioral skills. The IMB model provided an acceptable fit to the data (root mean square error of approximation < .05) and accounted for 50% of the variance in posttreatment condom use among the sample. Treatment effects on condom use were almost entirely mediated by the IMB variables; specifically, motivation and enactment of behavioral skills mediated the intervention's impact on condom use. CONCLUSIONS: These results provide supporting evidence as to how theoretical variables operate to effect change within a theory-based intervention and provide evidence as to the applicability of a prevailing theory of HIV risk behavior among low-income minority women. PMID- 16472042 TI - Chronic stress influences ambulatory blood pressure in adolescents. AB - BACKGROUND: High ambulatory blood pressure (ABP) predicts cardiovascular events, even after controlling for clinic BP and other established risk factors. PURPOSE: This study examined whether chronic or discrete stress in the past year was associated with greater ABP in adolescents. METHOD: Participants were 217 male and female Black and White adolescents who wore ABP monitors on 2 consecutive school days and completed a survey of life events. RESULTS: Report of discrete, negative events was not associated with ABP. Adolescents experiencing greater numbers of chronic, negative life events exhibited greater systolic blood pressure (SBP), independent of ethnicity; sex; body mass index; and location, position, physical activity, and consumption of food/caffeine/nicotine at time of measurement. Greater numbers of chronic, negative events were associated with greater diastolic blood pressure (DBP) among boys. When resting clinic BP was introduced into the model, the main effect of chronic, negative life events on ambulatory SBP became nonsignificant, whereas the effect of chronic, negative life events on male adolescents' DBP persisted. CONCLUSIONS: Chronic stress is associated with greater ambulatory SBP among male and female adolescents and with greater ambulatory DBP among male adolescents. The latter association persisted after controlling for clinic BP, suggesting that males may be more vulnerable to chronic stress as a determinant of BP regulation than females early in life. PMID- 16472043 TI - Modeling and rehabilitation following anterior cruciate ligament reconstruction. AB - BACKGROUND/PURPOSE: This study sought to examine the effectiveness of a modeling video to reduce preoperative perceptions of anxiety and pain, as well as to increase postoperative self-efficacy and functional outcomes after anterior cruciate ligament reconstruction. METHODS: Following baseline assessment of state anxiety, perceptions of expected pain, injury severity, and knee function (International Knee Documentation Committee [IKDC] system), patients scheduled for surgical reconstruction of the anterior cruciate ligament were randomly assigned to either a modeling intervention or a control group. Psychological assessments were repeated preoperatively for expected pain and anxiety. Actual pain was assessed preoperatively, prior to discharge, and at 2 weeks postoperatively. Rehabilitation self-efficacy was assessed prior to discharge and at 2 and 6 weeks postoperatively. IKDC functional assessments were repeated at 6 weeks postoperatively, whereas range of motion was assessed at 2 and 6 weeks postsurgery. RESULTS: Compared with the participants in the control condition, participants assigned to the modeling intervention reported significantly lower perceptions of expected pain preoperatively and significantly greater self efficacy at predischarge to perform rehabilitation tasks. Those who received the modeling intervention also experienced significantly better IKDC objective functional outcome scores compared with their control counterparts. No psychological variables mediated relations between the intervention and functional outcomes. CONCLUSIONS: The data suggest that watching a modeling video may be an effective prophylactic treatment to decrease perceptions of expected pain, increase rehabilitation self-efficacy, and provide an early stimulus with respect to early function. PMID- 16472044 TI - Physical activity and quality of life in older adults: influence of health status and self-efficacy. AB - BACKGROUND: Physical activity has been positively linked to quality of life (QOL) in older adults. Measures of health status and global well-being represent common methods of assessing QOL outcomes, yet little has been done to determine the nature of the relationship of these outcomes with physical activity. PURPOSE: We examined the roles played by physical activity, health status, and self-efficacy in global QOL (satisfaction with life) in a sample of older Black and White women. METHOD: Participants (N = 249, M age = 68.12 years) completed multiple indicators of physical activity, self-efficacy, health status, and QOL at baseline of a 24-month prospective trial. Structural equation modeling examined the fit of 3 models of the physical activity and QOL relationship. RESULTS: Analyses indicated that relationships between physical activity and QOL, self efficacy and QOL were all indirect. Specifically, physical activity influenced self-efficacy and QOL through physical and mental health status, which in turn influenced global QOL. CONCLUSIONS: Our findings support a social cognitive model of physical activity's relationship with QOL. Subsequent tests of hypothesized relationships across time are recommended. PMID- 16472045 TI - Inflammatory markers and type 2 diabetes. PMID- 16472046 TI - The epidemiology of low-grade chronic systemic inflammation and type 2 diabetes. AB - The fattening of the human species and the accompanying emergence of the metabolic syndrome and of type 2 diabetes as remarkably frequent clinical entities are among the major epidemiologic events of our time. Control of the diabetes epidemic requires a greater understanding of the pathophysiologic processes underlying these phenomena. Many epidemiologic studies have now shown associations between inflammation markers and diabetes, with the most consistent being for leukocytes and the strongest being for C-reactive protein. Consistent protective associations have also been reported for adiponectin, an adipocyte secretory protein with antiinflammatory actions. Although great variability is seen between reported associations, as a whole these studies suggest a role for inflammation linked to obesity. The variability reported is in part due to differences in model adjustment, in how diabetes was ascertained, and in the different means used to operationalize the concept of low-grade chronic systemic inflammation. It is also due, in part, to sample characterization, as findings are heterogeneous across some subgroups, such as those defined by smoking. Consistent with their association with type 2 diabetes, inflammation markers have also be shown to predict conditions present in the prediabetes state such as weight gain, hypertension, gestational diabetes, and decline in insulin sensitivity. PMID- 16472047 TI - "Anti-inflammatory" drugs and their effects on type 2 diabetes. AB - There is a growing body of evidence for the role of inflammation in type 2 diabetes. In addition to the evidence presented elsewhere, evidence is emerging that many drugs that have apparent "anti-inflammatory" properties may reduce the incidence and/or delay the onset of type 2 diabetes. Statins have been found to lower inflammatory markers, and a post hoc analysis of the West of Scotland Coronary Prevention Study (WOSCOPS) suggested that pravastatin may reduce the risk of developing diabetes, although the Lipid Lowering Arm of the Anglo Scandinavian Cardiac Outcomes Trial (ASCOT) found no statistically significant effect of atorvastatin on risk of developing diabetes. Fibrates have been found to lower some markers of inflammation, and a prospective trial found that bezafibrate reduces risk of developing diabetes. Angiotensin converting enzyme (ACE) inhibitors and angiotensin receptor blockers appear to reduce some markers of inflammation, and a meta-analysis concluded that ACE inhibitors and angiotensin receptor blockers reduce risk of developing type 2 diabetes. Metformin is known to reduce the risk of developing diabetes, and more recent evidence suggests it also lowers C-reactive protein, in part because of its modest weight-reducing effect. Thiazolidinediones reduce risk of developing diabetes, and consistently lower inflammatory markers independent of adiposity effects. High-dose aspirin inhibits cyclooxygenase and IkappaB kinase-beta and reduces fasting plasma glucose concentration, although there has not, as yet, been a large-scale trial to examine the effect of aspirin on the risk of developing diabetes. We conclude that although many drugs with potential anti inflammatory properties reduce the risk of developing diabetes, it is difficult to prove that such anti-inflammatory properties contribute to their diabetes prevention since nearly all drugs have other, often more pronounced, actions. Studies with more specific inhibitors of inflammatory pathways (e.g., interleukin 6 blockers) and mendelian randomization (genetic studies) will help determine whether targeting the inflammation axis is a fertile mechanism to treat or prevent type 2 diabetes. PMID- 16472048 TI - High-sensitivity C-reactive protein as cardiovascular risk marker in patients with diabetes mellitus. AB - C-reactive protein (CRP) is a liver-derived pattern recognition molecule that is increased in inflammatory states. It rapidly increases within hours after tissue injury, and it is suggested that it is part of the innate immune system and contributes to host defense. Since cardiovascular disease is at least in part an inflammatory process, CRP has been investigated in the context of arteriosclerosis and subsequent vascular disorders. Based on multiple epidemiological and intervention studies, minor CRP elevation [high-sensitivity CRP (hsCRP)] has been shown to be associated with future major cardiovascular risk (hsCRP:<1 mg/L=low risk; 1-3 mg/L=intermediate risk; 3-10 mg/L=high risk; >10 mg/L=unspecific elevation). It is recommended by the American Heart Association that patients at intermediate or high risk of coronary heart disease may benefit from measurement of hsCRP with regard to their individual risk prediction. Elevation of hsCRP is associated with increased risk of type 2 diabetes development in patients with all levels of metabolic syndrome. In type 1 and type 2 diabetes mellitus, hemoglobin A1c significantly correlates with hsCRP levels and future cardiovascular risk. Also, hsCRP levels increase with the stage of beta-cell dysfunction and insulin resistance. Non-diabetes drugs that have been shown to reduce hsCRP concentrations include aspirin, statins, cyclooxygenase-2 inhibitors, and fibrates. Recent intervention studies have also demonstrated the distinct efficacy of different anti-diabetes treatments on a variety of cardiovascular risk markers. Intensive insulin therapy may reduce inflammation, but this effect may be influenced by the degree of weight gain. Treatment with peroxisome proliferator-activated receptor gamma has lead to substantial reduction of hsCRP and other cardiovascular risk markers in several comparator studies. Since this effect was shown to be independent of the degree of glycemic improvement, it can be regarded as a classspecific effect. Whether these findings translate into a reduction of overall cardiovascular mortality will soon be shown by the currently running thiazolidinedione outcome studies. Positive results in these trials will further strengthen the value and acceptance of hsCRP, which is recommended as a predictive laboratory marker for cardiovascular disease risk also in patients with diabetes mellitus. PMID- 16472049 TI - An overview of inflammatory markers in type 2 diabetes from the perspective of the clinical chemist. AB - C-reactive protein (CRP), when measured by a highly sensitive method, is a measure of lowgrade, chronic inflammation and is an independent risk factor for type 2 diabetes (T2D) and cardiovascular disease (CVD). CRP also has the capacity to interact with other risk factors to increase the risk for T2D and CVD. Population distributions divided into tertiles provide the capacity to predict onset of T2D and associated CVD. Preanalytical as well as analytical sources of variation in high-sensitivity CRP (hsCRP) measurements need to be standardized in order for CRP results to be optimally useful. The Centers for Disease Control and Prevention and the American Heart Association have issued guidelines for clinical usefulness of hsCRP measurements. The Centers for Disease Control and Prevention has taken steps to standardize hsCRP assays by evaluating secondary reference materials to be used by manufacturers to calibrate their assays. PMID- 16472050 TI - Nutritional influences on inflammation and type 2 diabetes risk. AB - There is a clear role for inflammation in the development of type 2 diabetes and its associated co-morbidities. Circulating inflammatory markers such as C reactive protein, sialic acid, and interleukin- 6 are all significant independent predictors of disease. A number of nutritional components are hypothesized to modulate inflammation, and hence impact on disease risk. The most extensively studied nutrients are the long-chain n-3 polyunsaturated fatty acids. However, evidence is also emerging with respect to whole grain or low glycemic index foods and antioxidant vitamins. Obesity, resulting from long-term dietary energy excess, is also strongly linked to raised inflammatory status and type 2 diabetes. To date, much of the evidence for the effect of nutrients or foods on disease risk has been based on epidemiological associations. However, the links among diet, inflammation, and type 2 diabetes are supported by some data from human dietary intervention trials and/or mechanistic studies in animals. Further research is required to quantify the precise role and refine the evidence base. However, the proposed "anti-inflammatory" strategies to tackle type 2 diabetes are broadly consistent with current public health nutrition guidelines: to achieve and maintain a healthy weight, to reduce saturated fat, to increase the proportion of less refined forms of carbohydrate, and to increase intake of fruits and vegetables. PMID- 16472051 TI - Genetic predispositions to low-grade inflammation and type 2 diabetes. AB - Insulin resistance and cardiovascular disease share common pathophysiological mechanisms, as the chronic activation of the innate immune system. This system constitutes the first line of body's defense and is constituted by different barriers (e.g., epithelia, adipose tissue) and different blood and tissue components (e.g., macrophages, neutrophils). This system generates the acute phase response in which different acute-phase proteins and cytokines are produced in response to different aggressions as infections and traumatisms. The aim of this response is to eradicate these agents, to repair the harmed tissues, and, through increased insulin resistance, to optimize the energetic substrates, which will be drained to vital tissues and organs (i.e., brain and the immune system). Evolutionary pressures have led to survival of the fittest individuals, those with the genetics that allows the best defense against infection and periods of famine. Evidence is reported according to which gene polymorphisms in the molecules regulating the inflammatory cascade are associated with body composition, insulin action, and characteristics of the metabolic syndrome. The evolutive advantages of increased inflammatory responses, hypersecretion of proinflammatory cytokines [tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, IL-6, and IL-18], or decreased anti-inflammatory molecules (adiponectin, certain TNF-alpha isoforms, soluble CD14, etc.), would lead in westernized countries to chronic inflammation conditions, such as obesity and type 2 diabetes, resulting in cardiovascular disease. PMID- 16472052 TI - Early detection of insulin deprivation in continuous subcutaneous insulin infusion-treated patients with type 1 diabetes. AB - BACKGROUND: This study was performed to define the clinical relevance of early changes of capillary 3beta-hydroxybutyrate (3betaOHB), for detection of metabolic deterioration before occurrence of overt diabetic ketoacidosis following interruption of continuous subcutaneous insulin infusion (CSII). METHODS: An open clinical trial was performed with eight patients with type 1 diabetes on CSII therapy. After an overnight fast, at 8 a.m. (T0) CSII was interrupted for 4 h. At noon (T240) CSII was re-established, and at 4 p.m. (T480) the study was ended. Blood glucose (BG) and capillary and plasma 3betaOHB were measured at 30-min intervals, plasma insulin at 60-min intervals, and urinary ketones at 120-min intervals. RESULTS: After CSII interruption mean BG increased from 149.8+/-54.4 mg/dL at T0 to 224.8+/-56.2 mg/dL at T240 (P<0.05), and mean capillary 3betaOHB increased from 0.1+/-0.1 mmol/L at T0 to 0.9+/-0.6 mmol/L at T240 (P<0.001). The rate of increase of capillary 3betaOHB was faster and significantly more relevant than that of BG (P<0.05). The restoration of CSII produced a significant reduction of mean BG and capillary 3betaOHB (T480, 119.5+/-24 mg/dL and 0.2+/-0.2 mmol/L, respectively; P<0.05 for both vs. T240). The recovery of capillary 3betaOHB was significantly faster than that of BG (P=0.03). CONCLUSIONS: The dynamic evaluation of changes of capillary 3betaOHB levels can represent a useful support to home BG monitoring in the event of CSII interruption, providing faster information on early metabolic deterioration due to insulin deprivation and allowing preventative action for avoiding the evolution towards overt diabetic ketoacidosis. After reintroduction of insulin infusion the monitoring of the faster recovery of 3betaOHB relative to BG can provide useful information for the prevention of late hypoglycemia due to insulin overinfusion. PMID- 16472053 TI - Assessment of trueness of a glucose monitor using interstitial fluid and whole blood as specimen matrix. AB - BACKGROUND: Interstitial fluid (ISF) is a specimen of increasing interest for glucose measurements because it can be obtained in a minimally invasive manner. Our previous study showed that sufficient ISF can be obtained using microneedles to measure glucose with a conventional electrochemical glucose monitor. The aim of this study was to assess the trueness of this glucose monitor using split sample comparison with whole blood. We used ISF as specimen and our gas chromatography/mass spectrometry (GC/MS) method as reference. METHODS: We obtained 50 ISF samples and 40 whole blood samples from hairless Sprague- Dawley rats and analyzed for glucose by both methods. RESULTS: For whole blood, a non significant bias of 5.7% (+/-2 SD: -54.9% to 66.3%) was determined. ISF glucose measurements showed a significant constant bias of 29.5% (+/-2 SD: -85.0% to 144%), which seems to be caused in part by the lack of red blood cells in ISF. The correlation coefficients were 0.782 and 0.679 for whole blood and ISF, respectively. CONCLUSIONS: The assessed electrochemical glucose monitor shows a close agreement with our GC/MS reference method for whole blood, for which this monitor was optimized. When glucose measurements are performed with ISF as matrix, the observed bias needs to be taken into consideration. Further studies are necessary to elucidate the reasons for the wide dispersion of data for ISF. PMID- 16472054 TI - Pharmacokinetics and pharmacodynamics of intranasal insulin administered to patients with type 1 diabetes: a preliminary study. AB - BACKGROUND: The pharmacokinetics and pharmacodynamics of a Bentley Pharmaceuticals (Exeter, NH) proprietary insulin formulation designed for intranasal administration were studied in patients with type 1 diabetes, using subcutaneous insulin or placebo as comparator. METHODS: Seven fasting volunteer patients with type 1 diabetes (five men, two women; body mass index 23.54+/-1.32 kg/m2) received up to four doses of medication (placebo, subcutaneous insulin, or intranasal insulin) at least 3 days apart. Serum insulin and plasma glucose were measured in the 5-h period after dosing. RESULTS: The relative bioavailability of intranasal insulin compared with subcutaneous insulin was 16.6-19.8% over 2 h and 14.0-19.8% over 5 h. The formulation was generally well tolerated. At doses of 25 IU and above, a rise in serum insulin levels accompanied by a decrease in plasma glucose was seen. Peak insulin levels were generally attained in 15-20 min and remained elevated for approximately 1 h; the resultant effect upon glucose peaked at 40 min and waned 1.5-2 h post-dosing. The effect was dose related. Mean peak insulin levels increased with dose escalation. As reported in other insulin studies, the inter-individual responsiveness to insulin was variable. CONCLUSIONS: This intranasal formulation was generally well tolerated, and relatively well absorbed as demonstrated by a rapid rise in serum insulin level and concomitant reduction of plasma glucose levels. PMID- 16472055 TI - Initial clinical testing of a holographic non-invasive contact lens glucose sensor. AB - INTRODUCTION: The purpose of the present study was to determine the effectiveness of a new holographic contact lens glucose sensor for the non-invasive monitoring of blood glucose. METHODS: One fasting normal subject was given an oral challenge consisting of 44 g of glucose. The contact lens hologram signal and fingerstick blood glucose were measured over a 26- min period. RESULTS: The contact lens hologram signal appeared to track blood glucose well. The contact lens was comfortable and well tolerated. CONCLUSION: The holographic contact lens glucose sensor shows promise as a non-invasive home glucose monitor. PMID- 16472056 TI - Impact of environmental temperature on skin thickness and microvascular blood flow in subjects with and without diabetes. AB - BACKGROUND: Glucose measurement from different skin areas might be influenced by changes in skin texture due to several environmental confounders. Our study was performed to investigate the effect of changes in ambient temperature on skin thickness and microvascular skin blood flow in subjects with and without diabetes at the lower forearm. METHODS: Thirteen subjects with diabetes and seven without diabetes participated in the study. The investigations were performed in a temperature- and humidity-controlled climatic chamber (EMPA, St. Gallen, Switzerland). Starting at 25 degrees C, the environmental temperature was changed in 4 degrees C steps every 40 min. Skin thickness was measured by an ultrasonic reflection technique, and microcirculation was measured by laser Doppler fluxmetry at the lower forearm. Study participants underwent the entire procedure on up to four separate study trials. RESULTS: Our study revealed a significantly reduced skin thickness (P<0.05) and microvascular blood flow (P<0.05) in patients with diabetes mellitus compared with controls without diabetes during the entire investigation. During declining ambient temperature a significant reduction in skin thickness (with diabetes, -0.09+/-0.13 mm; without diabetes, -0.06+/-0.11 mm; P<0.05) and microvascular blood flow (with diabetes, -41+/-49 arbitrary units; without diabetes, -46+/-51 arbitrary units; P<0.05) could be observed in both groups without significant differences between the two. CONCLUSIONS: Although skin thickness and microvascular skin blood flow at the lower forearm were found to be reduced in patients with diabetes compared with controls without diabetes, both groups revealed comparable dynamics in skin thickness and microvascular blood flow during changes in environmental temperature. PMID- 16472057 TI - Accuracy and reliability of teleophthalmology for diagnosing diabetic retinopathy and macular edema: a review of the literature. AB - Using teleophthalmology for the delivery of routine eye care for patients with diabetes mellitus is becoming an increasingly common practice. Paramount in the consideration of any new diagnostic test is an analysis of its diagnostic accuracy and reliability and how that compares with conventional care. This review summarizes existing data on the diagnostic accuracy and reliability of teleophthalmology and conventional clinic-based eye care for detecting diabetic retinopathy and diabetes mellitus. The sensitivity of ophthalmoscopy for detecting diabetic retinopathy performed by eye care clinicians has varied widely, with point estimates ranging from 0% to 96%. Alternatively, specificity has been universally high. The sensitivity of teleophthalmology for detecting diabetic retinopathy has been shown to be comparable, if not better, than clinic based examinations. Sensitivity values have ranged from 50% to 93%. The specificity of teleophthalmology, like clinic-based examinations, has been consistently high. High levels of diagnostic reliability, analyzed by both simple agreement and kappa values, have been found between ophthalmoscopy and teleophthalmology for detecting and classifying diabetic retinopathy. Evaluating the accuracy of macular edema detection requires the use of dual gold standards, the clinical examination using slit-lamp biomicroscopy and stereoscopic photography. Teleophthalmology, compared with both gold standards, has, overall, been a highly sensitive and specific test. Reliability studies that compared the two gold standards with one another have found moderate to substantial levels of agreement. Based on existing data, teleophthalmology appears to be an accurate and reliable test for detecting diabetic retinopathy and macular edema. PMID- 16472058 TI - A hydrogel-based implantable micromachined transponder for wireless glucose measurement. AB - In this paper, we report on the design and characterization of a new hydrogel based implantable wireless glucose sensor. The basic device structure is a passive [inductor/capacitor (LC)] micromachined resonator coupled to a stimuli sensitive hydrogel, which is confined between a stiff nanoporous membrane and a thin glass diaphragm. As glucose molecules pass through the nanoporous membrane, the hydrogel swells and deflects the flexible glass diaphragm, which is the movable plate of the variable capacitor in the totally integrated passive LC resonator. The corresponding change in resonant frequency can be remotely detected. A glucose- sensitive phenylboronic acid-based hydrogel was loaded into the microtransponder, and its sensitivity and time response were measured. Prior to hydrogel loading, the sensitivity of the pressure sensor to applied air pressure was measured to be -222 kHz/kPa over the frequency range 51-->42 MHz. The sensor showed a sensitivity of -34.3 kHz/mM over the glucose concentration range 0-20 mM (at pH 7.4), and a response time of 90 min. The dynamic response, although unacceptable at such values, can be easily improved by decreasing the hydrogel thickness and reducing the sensor and porous membrane thicknesses. The transponder's overall dimensions were 5x5x0.8 mm3, small enough for subcutaneous implantation. PMID- 16472059 TI - Analysis: biomedically enabled soldiers: super-resilient, not superhuman. PMID- 16472060 TI - Interactive educational diabetes/insulin tutorial at www.2aida.info. AB - The World Wide Web now hosts a multitude of diabetes educational materials in various formats. Of particular interest is the diabetes/insulin tutorial available at the AIDA Website (accessible directly at: www.2aida.info). The tutorial combines textual or "static" information with an interactive diabetes simulator-AIDA online-to provide an engaging and effective educational tool. AIDA online (accessible directly at: www.2aida.net) enables the simulation of plasma insulin and blood glucose levels from user-defined insulin injection and carbohydrate intake data. A haemoglobin A1c value is also computed, giving an indication of overall blood glucose control in the virtual patient with diabetes. The diabetes/insulin tutorial is currently composed of four sections: the first two cover in considerable depth insulin injection regimens and insulin dosage adjustment; the third section introduces the principles of carbohydrate counting and, specifically, matching insulin doses to carbohydrate intake; and the fourth section illustrates the relationship between blood glucose levels and renal excretion of glucose. The simulator runs alongside the tutorial, and allows various concepts described in the text to be explored freely by the user and simulated interactively. This introduces a novel way of learning how injected insulin and dietary carbohydrate interact in various insulin injection regimens. A fifth section- for which any offers of assistance would be gratefully received is planned. This will consider the use of insulin pumps and rapidly acting and very long-acting insulin analogues. Further improvements that may strengthen the existing tutorial and/or use of the online simulator are discussed in this column. PMID- 16472061 TI - Linkage of lamins to fidelity of gene transcription. AB - Two major events are emerging as being involved in the fine regulatory mechanisms that control gene expression through the nuclear envelope/lamina. The first is the linkage of envelope proteins to transcription factors, most of which function as transcriptional repressors. The second is the interplay between nuclear lamina/nuclear envelope proteins and chromatin, through more or less direct interactions with DNA-binding proteins. The first mechanism appears to play a major role in tissue-specific functions, whereas the latter mechanism likely modifies higher-order chromatin organization, thus regulating differentiation and aging. This review highlights the importance of the study of a group of genetic diseases, collectively referred to as laminopathies, to understand the actual role played by lamins in ensuring the fidelity of gene expression and in modulating cell differentiation. PMID- 16472062 TI - Measuring DNA nucleobase adducts using neutral hydrolysis and liquid chromatography-mass spectrometry. AB - The detection and identification of DNA adducts is important for predicting human cancer risk posed by chemicals and for uncovering potential genotoxicity of new drug and agricultural chemical candidates. For compounds that react with DNA to form N7-guanine and/or N3-adenine adducts, neutral thermal hydrolysis provides a simple procedure for sample preparation. The N7-guanine and N3-adenine adducts are selectively ejected from the DNA chain, resulting in a clean sample matrix enriched in nucleobase adducts. Coupling neutral thermal hydrolysis with liquid chromatography-mass spectrometry (LC-MS) provides sensitive methods to detect and quantitate DNA adducts, and structural information is provided by MS. Combining these technologies with capillary liquid chromatography sample preconcentration systems can provide exquisitely sensitive detection. In this review, we first summarize the chemistry of nucleobase adduct formation, briefly summarize modern methods to detect DNA adducts, and then describe neutral thermal hydrolysis coupled to LC-MS/MS and some of its applications to DNA damage studies. Finally, we review recent applications of neutral thermal hydrolysis and LC-MS to toxicity screening of chemicals. PMID- 16472063 TI - Polymeric gene carriers. AB - Polymeric gene carriers are a potential alternative to using viral vectors. Polymeric carriers have relatively low immunogenicity and cytotoxicity. In addition, polymeric carriers can accommodate large-size DNA, be conjugated with appropriate functionalities, and be administered repeatedly. In spite of these advantages, polymeric gene carriers have some limitations, such as low gene transfection efficiencies and relatively short duration of gene expression. Therefore, extensive research has been conducted toward the development of efficient polymeric carriers. In this review, we discuss current problems associated with polymeric gene carriers and various strategies against transfection barriers in particular, gene stabilization and protection, cellular targeting, endosomal escaping, nuclear targeting, unpackaging, and biocompatibility. Finally, requirements for future polymeric gene carriers are considered. With all these ongoing efforts, polymeric carriers have become one of the promising gene delivery methods for human gene therapy. PMID- 16472064 TI - Age-dependent biomechanical modifications in bone. AB - Age-related nontraumatic fractures are a major public health problem. Even though lower bone mass is the most commonly implicated variable for the age-related increase in fracture incidence, studies show that the resistance of bone material against fracture (toughness) diminishes with age. The mechanisms for the age related loss of toughness are, however, unknown and may involve alterations in the quality of the bone material. The review of literature presented here identifies changes in the quality of bone material at ultrastructural, lamellar, osteonal, and tissue levels as plausible contributors to the biomechanical changes in aging human bone. The changes in bone quality at these multiple levels of organization result in a systematic breakdown of the mechanical and biological mechanisms that provide bone with its resistance against fracture. Animal studies conducted to date suggest that, similar to bone mass, bone quality may be partially regulated by genetics. PMID- 16472066 TI - MHC transfer from APC to T cells following antigen recognition. AB - Recognition of cognate MHC:peptide complexes by T cells leads to large-scale molecular rearrangements resulting in immunological synapse formation at the T cell-antigen-presenting cell (APC) interface. Although the functions of the immunological synapse are not completely understood, a consequence of this event appears to be the intercellular transfer of MHC:peptide complexes, along with other molecules such as CD80, from the APC to the T cell. The expression of APC derived molecules on the T cell is biologically significant. It has the potential to alter the homing, allow T cells to also act as APC, and may alter the effector functions of the cell. Experimental evidence suggests that intercellular transfer may play a role in the control of an immune response; however, the exact role is unclear. Both potentiation and attenuation of an ongoing response have been postulated. In addition, removal of molecules from APC may be important in controlling homeostatic proliferation, in affinity maturation of T cells, and in maintaining epitope diversity during an immune response. In this review, we highlight recent advances regarding the mechanism of intercellular transfer and focus on the potential biological significance of this event. PMID- 16472067 TI - Recombination in the human IGK locus. AB - The human IGK locus is probably the most complex of all antigen-receptor loci because of the presence of many elements that can be involved in V(D)J recombination. Next to classical Vkappa-Jkappa recombinations and deletional recombinations involving Kde (intron-Kde, Vkappa-Kde), JkappaRSS-intronRSS signal joints and Vkappa-intronRSS and JkappaRSS-Kde pseudohybrid joints are observed in human B-cell malignancies and also in normal human B cells. These recombinations occur in an ordered and tightly regulated way. Although DNA (de)methylation, histone (de)acetylation, and nuclear positioning all play a role in this regulation, much is still to be learned about the exact factors involved. One of these factors seems to be E2A that is known to be important in B-cell development and has a direct role in the induction of human endogenous IGK recombination. Contrary to the human IGH locus, aberrant recombinations resulting in chromosomal translocations are not very frequent in the human IGK locus. Despite this, IGK recombinations have diagnostic and clinical implications, being important PCR targets for clonality and MRD studies. PMID- 16472069 TI - Immune defects in Fanconi anemia. AB - Fanconi anemia (FA) is a genetic disorder characterized by sensitivity to DNA cross-linking agents, multiple congenital anomalies, progressive bone marrow failure, and an increased prevalence of malignancy. The nature of chromosomal instability in FA is better understood today than in the past, but the molecular pathogenesis of bone marrow failure in this disease has not been clarified. Although there is documented evidence that FA hematopoietic stem cells (HSC) have inherent defects that reduce their survival, the potential influence of auxiliary cells on the ability of the FA bone marrow microenvironment to maintain and support HSC in unknown. Historically, FA has not been represented as a disease that affects the lymphoid compartment. In this article we review the results of studies that suggest that the FA immune system is dysfunctional and may contribute to the pathogenesis of both FA bone marrow failure and neoplastic disease. PMID- 16472068 TI - Cell surface glycans: the why and how of their functionality as biochemical signals in lectin-mediated information transfer. AB - Calculations of the coding capacity of biomolecules come up with the striking result that carbohydrates excel nucleotides and amino acids by far in this respect. Consequently, glycans are ideally suited to equip cell surfaces with a maximum of signals in a minimum of space and form the cellular glycome. The ensuing concept of the sugar code interprets glycans as a platform for encoding messages and shifts in the glycome as a means to swiftly add/remove signals. Next, it implies the presence of effectors that read the signals in situ. Fittingly, recent work on natural decoding devices in mammals (endogenous lectins) has revealed not only their presence but a level of complexity that matches the structural diversity of glycans. The emerging importance of lectin ligand interaction in immune regulation calls for an introduction to this concept for nonspecialists. This primer to the fundamentals of the sugar code, with telling examples from the realm of immunology, also illustrates the strategic and hitherto unsuspected fine-tuning between glycan structure and lectin design/expression. PMID- 16472070 TI - Autoantibody reaction to myelin basic protein by plasma parvovirus B19 IgG in MS patients. AB - The etiology of multiple sclerosis (MS) remains unclear. To determine if autoantibodies to myelin basic protein (MBP) are produced during parvovirus B19 infection, a competitive ELISA was performed using plasma from MS patients exhibiting high IgG titers for parvovirus. Our results showed the addition of MBP decreased the binding of IgG to B19 antigen in a dose dependent fashion suggesting a possible link between parvovirus B19 and a subset of patients with clinical MS. PMID- 16472071 TI - Effect of a new ionic pair on the unfolding activation barrier of beta glucosidase B. AB - Thermal unfolding kinetics of beta-glucosidase B from Paenibacillus polymyxa and its thermoresistant mutant H62R were determined from far-UV circular dichroism (CD) measurements at different temperatures. The unfolding of both enzymes followed simple two-state kinetics. The new ionic pair formed between Arg62 and Glu429 in the H62R variant did not change substantially the enzyme structure as judged by far-UV CD and fluorescence spectra, but produced an increase in the unfolding activation barrier of 0.95 +/- 0.10 kcal mol(-1), in good agreement with the energetic contribution reported for surface salt bridges in proteins. Eyring's analysis of the unfolding kinetic constants showed that the activation enthalpies for thermal denaturation of both enzymes were essentially the same. Thus, the greater kinetic stability rendered by the salt bridge seems to be due to a reduction in the activation entropy. PMID- 16472072 TI - Processing of amyloid beta-peptides by neutral cysteine protease bleomycin hydrolase. AB - We studied the processing of amyloid beta-peptides (Abetas) including Abeta(1 40), Abeta(1-42) and pAbeta(3-42) by rat neutral cysteine protease bleomycin hydrolase (BH) according to the methods of SDS-PAGE, HPLC and matrix-assisted laser desorption/inonization time-of-flight mass spectrometry (MALDI-TOF MS). BH significantly processed them by novel features of its diverse activities. It initially cleaved at two sites, His(14)-Gln(15) and Phe(19)-Phe(20) degraded to short intermediates then to amino acids by aminopeptidase and/or carboxypeptidase activities. Also, full-length Abetas were clipped at the carboxyl(C)-terminal region. On the other hand, BH cleaved at only the His(14)-Gln(15) bond in pbetaA(3-42) within a short period of the reaction by endopeptidase activity, and processed the intermediates in order by carboxypeptidase activity. On processing by BH, it found that both fibrillar Abeta(1-40) and Abeta(1-42) were more resistant than non-fibrillar peptides. These results indicate that the processing specificity of BH depends upon the structure and sequence of Abetas. PMID- 16472073 TI - A nonradioactive assay for poly(a)-specific ribonuclease activity by methylene blue colorimetry. AB - A simple nonradioactive assay, which was based on the specific shift of the absorbance maximum of methylene blue induced by its intercalation into poly(A) molecules, was developed for poly(A)-specific ribonuclease (PARN). A good linear relationship was found between the absorbance at 662 nm and the poly(A) concentration. The assay conditions, including the concentration of methylene blue, the incubation temperature and time, and the poly(A) concentration were evaluated and optimized. PMID- 16472074 TI - Mitochondria-regulated death pathway mediates (DIPP-L-Leu)2-L-LysOCH3-induced K562 cells apoptosis. AB - (DIPP-L-Leu)2-L-LysOCH3 is a diisopropylphosphoryl dipeptide which is known to induce apoptosis of human leukemia K562 cells. The molecular and cellular mechanisms involved in this process remain to be clarified. Herein, we show that (DIPP-L-Leu)2-L-LysOCH3-induced apoptosis is associated with cytosolic accumulation of cytochrome c, sustained loss of mitochondrial transmembrane potential (MMP), transient generation of reactive oxygen species (ROS) and elevation of intracellular Ca2+ concentration. A specific caspase assay reveals an increase in caspase-9 and caspase-3 activity but no change in caspase-8 activity. Immunofluorescence analysis indicates that (DIPP-L-Leu)2-L-LysOCH3 induced upregulation of pro-apoptotic Bax and downregulation of anti-apoptotic Bcl-2 and Bcl-x(L). These results suggest that the mitochondria-regulated death pathway mediates (DIPP-L-Leu)2-L-LysOCH3-induced K562 cells apoptosis. PMID- 16472075 TI - Synthesis, characterization and biological activity of chemically modified insulin derivative with alpha lipoic acid. AB - A novel chemically-modified insulin, epsilon-N(B29)-lipoyl insulin, was selectively prepared by the covalent linkage of alpha-lipoic acid (LA) to the epsilon-amino group of Lys(B29) of insulin without any protecting agent and analyzed by PAGE, HPLC, MALDI-TOF-MS. Monolipoyl- insulin maintained the glucose lowering effect as well as native insulin and showed a longer duration of action than native insulin and an inhibitory effect towards trypsin degradation. PMID- 16472076 TI - Timing of mutation in influenza A virus hemagglutinins by means of amino-acid distribution rank and fast Fourier transform. AB - In this study, we calculated the amino-acid distribution rank of 1201 hemagglutinins from influenza A viruses dated from 1918 to 2004 in order to compare them with respect to subtypes, species and years. After noticing fluctuations in distribution rank along the time course, we used the fast Fourier transform to determine the mutation periodicity of the hemagglutinins. Then we estimated our position at the current cycle of hemagglutinin evolutionary process to determine how many years remain before the next possible outbreak of influenza and bird flu. Finally, we used the trend channel to outlook the future of hemagglutinins for the next half a century. As our study covers almost all the full-length amino-acid sequences of hemagglutinins from various influenza A viruses, the conclusions will be valid for years until the number of hemagglutinins in Protein Databank is significantly increased. PMID- 16472077 TI - Asparaginase display of human cholesteryl ester transfer protein (CETP) B cell epitopes for inducing high titers of anti-CETP antibodies in vivo. AB - The recombinant chimeric enzyme, AnsB-TTP-CETPC, comprising asparaginase, tetanus toxin helper T cell epitope and human CETP B cell epitope was expressed as a soluble protein in Escherichia coli. The purified chimeric enzyme exhibited approximate 83% activity of the native asparaginase. After immunization with three doses of chimeric enzyme, high titers of anti-CETP antibodies were induced and lasted more than eighteen weeks in mice, and could even be detected at a dilution of 1:12800 by normal ELISA assay. The specificity of anti-CETP antibody was verified by Western blot assay. After displaying on the surface of asparaginase, the weak antigenicity of CETP epitope was effectively overcome, there after a strong CETP-specific immune response was evoked in mice immunized with the chimeric enzyme. Histochemical analysis of mice kidney tissue showed that immunization with the chimeric enzyme did not cause any pathological changes in mice. Collectively, the chimeric enzyme may be further developed as a vaccine against atherosclerosis in the future. PMID- 16472078 TI - Lys70 of E. coli quinolinate phosphoribosyltransferase is protected from chemical modification by formation of an inhibitor complex. AB - Quinolinate phosphoribosyltransferase was examined for susceptibility to different chemical modification reagents. Loss of enzyme activity with trinitrobenzenesulfonate (TNBS) occurred when 1.1 lysines per subunit were modified. Tryptic digestion of the modified enzyme followed by HPLC-MS analysis of the peptides showed Lys70 reacts with TNBS. Based on x-ray studies, this amino acid participates in a conformational change distant from the active site. PMID- 16472079 TI - Influence of human B-casomorphin-7 on specific binding of 3H-spiperone to the 5 HT2-receptors of rat brain frontal cortex. AB - Using radio-receptor analysis, it has been demonstrated that human beta casomorphin-7 (Tyr-Pro-Phe-Val-Glu-Pro-Ile) displaces 3H-spiperone from 5-HT2 receptors of rat brain frontal cortex. IC50 of human beta-casomorphin-7 was 8 microM. These data suggest that one of the mechanisms of neurotropic action of beta-casomorphin-7 is might be associated with its influence on the serotoninergic system. PMID- 16472080 TI - Identification and characterization of Trichoplusia ni furin truncated mutant. AB - We report a new prohormone convertase gene of insect origin, Trichoplusia ni furin, which was cloned from BTI-Tn-5B-4 (Tn5) insect cells. We constructed a truncated mutant that lacked Cys-rich repeated segments. Using baculovirus expression system and standard enzymatic assay, we obtained recombinant Tn furin and evaluated aspects of its function. PMID- 16472081 TI - Structure-based stabilization of an enzyme: the case of penicillin acylase from Alcaligenes faecalis. AB - The modeled structure of penicillin acylase from Alcaligenes faecali (AFPGA) was constructed by comparative modeling with the Modeller program. Candidate positions that could be replaced with cysteine were estimated by scanning the modeled structure of AFPGA with the program MODIP (modeling disulfide bond in protein). The mutant Q3C/P751C had a higher optimum temperature by three degrees than that of the wild type AFPGA. The half life of the double mutant Q3C/P751C at 55 degrees C was increased by 50%. To our knowledge, this was the first structure based genetic modification of AFPGA. PMID- 16472082 TI - Effects of [Nphe1]nociceptin(1-13)NH2, [Phe1(CH2-NH)Gly2]nociceptin(1- 13)NH2, and nocistatin on nociceptin inhibited constrictions of guinea pig isolated bronchus. AB - Electric field stimulation (EFS) causes excitatory non adrenergic-non cholinergic (eNANC) and cholinergic constrictions in the guinea pig isolated bronchus, the activation of eNANC and cholinergic nerves respectively. We investigated the effects of [Nphe1]nociceptin(1-13)NH2 ([Nphe1]NC(1-13)NH2), [Phe1(CH2 NH)Gly2]nociceptin(1- 13)NH2 ([F/G] NC(1-13)NH2), and nocistatin (NST) on nociceptin (NC) inhibited constrictions in isolated bronchus of guinea pig. The results show that NC (1 micromol/L) inhibited EFS-induced eNANC and cholinergic constrictions compared with the control, in which nociceptin was not applied. After pretreatment with [Nphe1]NC(1-13)NH2, [F/G]NC(1-13)NH2, or NST, the inhibitions of NC were antagonized by [Nphe1]NC(1-13)NH2 and [F/G]NC(1-13)NH2 but not NST. However, [Nphe1]NC(1-13)NH2, [F/G]NC(1-13)NH2, and NST did not affect the inhibitions induced by morphine. Furthermore, [Nphe1]NC(1-13)NH2, [F/G]NC(1 13)NH2 and NST did not cause any appreciable effects on EFS-induced eNANC and cholinergic constrictions in guinea pig bronchi. The results demonstrate that [Nphe1]NC(1-13)NH2 and [F/G]NC(1- 13)NH2 but not NST act as selective antagonists of the NC receptor and the effects of NC on EFS-induced constrictions of guinea pig isolated bronchus. PMID- 16472083 TI - Solid phase peptide synthesis in water VI: evaluation of water-soluble coupling reagents for solid phase peptide synthesis in aqueous media. AB - Solid phase peptide synthesis requires large amounts of organic solvents, the safe disposal of which is an important environmental issue. Peptide synthesis, if performed in water and using less or nontoxic reagents, circumvents the disposal problem. Our ultimate aim is to develop an "environment-friendly" solid phase peptide synthesis (SPPS) methodology. Previously, we showed that SPPS in water is feasible. To perform SPPS in water, the coupling reagent must be water-soluble and maintain its reactivity in water. For this report, we tested the efficacy of the water-soluble coupling reagents, 2-(5-norbornene-2,3-dicarboximido)-1,1,3,3 tetramethyluronium tetrafluoroborate (TNTU) and 4-(4,6-dimethoxy-1,3,5-triazin-2 yl)-4-methylmorpholinium chloride (DMT-MM), towards SPPS in water. We successfully synthesized Leu-enkephalin amide on a solid support suspended in aqueous 50% EtOH using DMT-MM and 2-(4-sulfophenylsulfonyl)ethoxycarbonylamino acids. PMID- 16472084 TI - Eph/Ephrin membrane proteins: a mammalian expression vector pTIg-BOS-Fc allowing rapid protein purification. AB - There is an urgent need for high purity, single chain, fully functional Eph/ephrin membrane proteins. This report outlines the pTIg-BOS-Fc vector and purification approach resulting in rapid increased production of fully functional single chain extracellular proteins that were isolated with high purity and used in structure-function analysis and pre-clinical studies. PMID- 16472085 TI - Expression and purification of active WW domains of FBP11/HYPA and FBP28/CA150. AB - Production of GST-fused WW domains of FBP proteins was increased using the bubbling cultivation method for E. coli. Purified WW domains of FBP11 and FBP28 bound a PL motif peptide with dissociation constants (K(D)) of 248 +/- 27 and 1880 +/- 280 microM, respectively. PMID- 16472086 TI - Expression, purification, crystallization and preliminary X-ray analysis of human spindlin1, an ovarian cancer-related protein. AB - Human spindlin1 is a newly screened and identified gene product related to ovarian carcinomas and is highly homologous to mouse spindlin. It is an abundant maternal transcript expressed in the mouse during the transition from oocyte to embryo. Here, the recombinant human spindlin1 has been overexpressed in Escherichia coli BL21, purified and crystallized using the hanging-drop vapour diffusion method. Crystals diffracting to 2.25 A resolution were obtained using ammonium sulfate as precipitant. The crystals belong to the space group P2(1)2(1)2(1), with unit-cell parameters a =40.7 A, b =84.4 A, c =136.4 A, alpha=beta=gamma=90 degrees . Assuming two molecules per asymmetric unit, the solvent content is calculated to be 42.4%. PMID- 16472087 TI - Crystallization and preliminary diffraction studies of malate dehydrogenase from Streptomyces aureofaciens. AB - Purified malate dehydrogenase (MDH) of Streptomyces aureofaciens was crystallized either in the absence or in the presence of NADH or NADPH coenzymes by hanging drop vapour-diffusion method. An X-ray study has shown, that MDH crystals belong to space group C222(1) with unit-cell parameters a = 53.2 A, b = 104.6 A, c = 520.0 A, alpha = beta = gamma = 90( degrees ), MDH-NADH crystals to space group C2 with unit-cell parameters a = 51.5 A, b = 51.5 A, c = 256 A, alpha = beta = gamma = 90( degrees ), and MDH-NADPH crystals to space group C222(1) with unit cell parameters a = 72, A b = 72 A, c = 520 A, alpha = beta = gamma = 90( degrees ). The crystal of native MDH diffracted to 2.1 A resolution. PMID- 16472089 TI - Novel methods for the non-invasive administration of DNA therapeutics and vaccines. AB - To develop a successful protein therapeutic, effective DNA delivery technologies are required that induce high and sustained levels of protein production in appropriate targets sites, whereas robust and long-lasting immune responses need to be induced by a DNA-based vaccine. Vectors for gene therapy and DNA vaccines must be resistant to degradation and attack by the immune system, have a satisfactory safety profile, and be able to express the therapeutic protein for the desired period of time. Effective non-viral vectors, which can express the proteins of interest at high levels, are available. However, since most of the DNA delivered in vivo is degraded before it can enter the nucleus, proper formulation and delivery are critical to the development of effective gene-based therapeutics and vaccines. These systems must be safe for human and veterinary clinical applications and yet ensure that the DNA survives the extra- and intracellular environment and is capable of entering the appropriate cellular compartments. In this review various potential and proven non-invasive chemical, mechanical, physical and biological DNA delivery systems for therapeutic and vaccine applications are discussed. A few of these approaches have been evaluated and proven to be promising in target species. Others, which promise to be less invasive, have only just started to be explored. PMID- 16472090 TI - DNA delivery for vaccination and therapeutics through the skin. AB - Cutaneous gene therapy and DNA vaccination are potential applications of plasmid delivery methods where a gene for an antigen or a therapeutic protein is inserted in the plasmid and applied to the skin. However, the delivery of the DNA plasmid is a major challenge due to the unusual physicochemical properties of the DNA, the tissue and cellular barriers and expression difficulties. Even though the skin is the most accessible organ of the body and it is an ideal target for gene therapy, the delivery of plasmid DNA across the skin is very difficult due to the specific barrier function of the stratum corneum and the inconsistent transfection rate of keratinocytes and other epidermal cells. To date there is no gene delivery system that was shown to be optimal for cutaneous gene therapy. In order to develop an efficient non-viral delivery vehicle we need to design a system that provides the combined properties of effective DNA condensation, cutaneous permeation, cellular transfection and sufficiently sustained expression. This paper reviews the formulation approaches and delivery methods for DNA through the skin in the context of the barriers both at the tissue and cellular levels for both vaccine and gene therapy applications. PMID- 16472091 TI - Non-invasive immunization on the skin using DNA vaccine. AB - Skin has evolved to protect not only by acting as a physical barrier, but also by its role in our powerful immune system. As a frontline of the host's defense against pathogens, skin is well equipped for immune surveillance. For example, compared to many other tissues, the epidermis of the skin contains a high population of Langerhans cells, which are very potent immature dendritic cells. Thus, targeting antigens to the skin epidermis should be able to efficiently induce strong immune responses. However, the forbidden barrier posed by the stratum corneum layer of the epidermis prevents effective entrance of antigens into the epidermis. Nevertheless, non-invasive immunization onto the skin has proven in the last several years to be a viable immunization modality. DNA vaccine is a vaccine made of bacterial plasmid DNA encoding an antigen of interest. Upon uptake of the plasmid, host express and process the encoding antigen, and then mount immune responses against it. DNA vaccine is advantageous over many other types of vaccines. The feasibility of non-invasive immunization onto the skin with DNA vaccine has been confirmed. Although the potency of the immune response has proven to be weak, many skin stratum corneum disrupting chemical and physical approaches and DNA vaccine carriers/adjuvants that significantly enhance the resulting immune response have been reported. In addition, research on elucidating the mechanism of immune induction from non invasively, topically applied DNA vaccine has also been carried out. With further improvement and optimization, non-invasive immunization onto the skin with DNA vaccine should be able to elicit reliable and efficacious immune response to a variety of antigens. PMID- 16472092 TI - Topical DNA vaccination with DNA/Lipid based complex. AB - Topical DNA vaccines have been shown to elicit both broad humoral and cellular immune response in vivo. The skin is an attractive site for the delivery DNA antigens for DNA vaccination. However, due to skin's barrier properties, the penetration of DNA and the applications of topical vaccination are limited. To improve permeability of stratum corneum and the potency of topical DNA vaccines, efficient delivery systems are needed. Topical vaccination has been achieved using topical application of naked DNA with or without tape stripping and DNA/lipid based complex such as liposomes, niosomes, Transfersomes, or microemulsion. All methods resulted in significant enhancement in humoral and cellular immune response over naked DNA alone. To develop more cost-effective and needle free vaccines, skin targeted immunizations are required. This overview focuses on the comparison of the potency of topical DNA vaccine between naked DNA and DNA-lipid based complex. PMID- 16472093 TI - Bacterial invasin: structure, function, and implication for targeted oral gene delivery. AB - The mucosal surface of the gastrointestinal (GI) tract is the first line of defense against foreign pathogens and toxins ingested orally. The content of the GI tract is constantly being sampled by the immune system through specialized epithelial cells known as M-cells, which are present in the Peyer's patches of the gut, providing a thin covering over lymphoid tissue. In this way, once a harmful entity is found an immune response can be activated to eliminate the threat. Many bacterial pathogens, such as Yersinia, Listeria, Salmonella, and Shigella, have evolved ways of exploiting M-cells to gain entrance to the body. The Yersinia species is of particular interest since its extracellular protein invasin provides one of the most direct and efficient manners of host cell invasion. Invasin binds to a subset of beta1 integrin receptors located on the apical membrane of intestinal M-cells, thereby facilitating the bacteria's entry into the cells and the lymphatic system underneath. This mechanism is highly specific and effective, making the invasin protein a very attractive modality for use in the oral delivery of molecules that include therapeutic genes and gene based vaccines. This article provides a brief overview of the molecular structure and properties of the Yersinia invasin as related to the protein's ability to facilitate binding and entry into M-cells. Also discussed are several innovative approaches that demonstrate the use of invasin as an effective targeting agent for biological and synthetic gene carrier systems, and the future prospect of developing invasin-based oral gene delivery formulations. PMID- 16472094 TI - Advances in noninvasive pulmonary gene therapy. AB - One of the most noninvasive approaches to drug delivery is via inhalation. The delivery of genes via aerosol holds promise for the treatment of a broad spectrum of pulmonary disorders and offers numerous advantages over more invasive modes of delivery. Delivery of genes expressing secretory therapeutic proteins or peptides may even have application to a number of nonpulmonary diseases. After the cloning of the cystic fibrosis gene, there was great interest in the delivery of genes directly to the lung surfaces via inhalation and most early efforts focused on the use of nonviral vectors, particularly cationic lipids. Early on, nebulization shear forces, inefficient penetration of mucous barriers and inhibitory effects of surfactant and other lung specific features generally resulted in a lack of therapeutic effect. But in recent years, a number of other nonviral and even viral vectors have been delivered successfully in this manner. Polyethyleneimine (PEI)-based formulations have proven stable during nebulization and result in transfection of a very large proportion of epithelial cells throughout the airways (though the level of transgene expression per cell may be relatively low), as well as significant, though lower levels of transfection throughout the lung parenchyma. Most importantly, therapeutic responses have been obtained in several animal lung tumor models when PEI-based complexes of p53 and IL-12 genes were delivered by aerosol. This approach may also prove useful as a means of localized genetic immunization. In addition, inhalation delivery of some formulations seems to be associated with surprisingly low toxicity and has resulted in little or no immunostimulatory response to the unmethylated CpG sequences in bacterially-produced plasmid DNA, which has presented a challenge to repeated gene therapy via many other modes of delivery. PMID- 16472096 TI - Sequence and time dependence of transfection efficiency of electrically-assisted gene delivery to tumors in mice. AB - Electrically-assisted gene delivery is a non-viral gene delivery technique, using application of square wave electric pulses to facilitate uptake of plasmid DNA into the cells. Feasibility and effectiveness of this method in vivo was already demonstrated, elaborating on pulse parameters and plasmid construction. However, there were no studies performed on sequencing and timing of plasmid DNA injection into the tumors and application of electric pulses. For this purpose we measured luciferase expression in two tumor models (LPB fibrosarcoma, B16F1 melanoma) after electrically-assisted gene delivery at varying time intervals between the pCMV-Luc plasmid injection and electroporation. Expression of luciferase was determined by measurement of its activity using luminometer. The results demonstrated that pCMV-Luc plasmid has to be injected before the application of electric pulses, since no measurable expression was detected in the tumors when pCMV-Luc plasmid was injected after electroporation of tumors. In both tumor models the highest transfection efficiency was obtained when pCMV-Luc plasmid was injected not less than 5 minutes but also not more than 30 minutes before the application of electric pulses. The results also demonstrated variability in the transfection efficiency depending on the tumor model. High expression was obtained in B16F1 tumor model (approximately 5500 pg luc/mg tumor) and lower in LPB fibrosarcoma (approximately 200 pg luc/mg tumor). In conclusion, our results demonstrate that regardless of the susceptibility of the tumors to electrically assisted gene delivery, the best timing for pCMV-Luc plasmid is between 30 to 5 minutes prior to the application of electric pulses to the tumors. PMID- 16472095 TI - Minimally invasive cutaneous delivery of macromolecules and plasmid DNA via microneedles. AB - The stratum corneum (SC) represents a significant barrier to the delivery of gene therapy formulations. In order to realise the potential of therapeutic cutaneous gene transfer, delivery strategies are required to overcome this exclusion effect. This study investigates the ability of microfabricated silicon microneedle arrays to create micron-sized channels through the SC of ex vivo human skin and the resulting ability of the conduits to facilitate localised delivery of charged macromolecules and plasmid DNA (pDNA). Microscopic studies of microneedle-treated human epidermal membrane revealed the presence of microconduits (10-20 microm diameter). The delivery of a macromolecule, beta galactosidase, and of a 'non-viral gene vector mimicking' charged fluorescent nanoparticle to the viable epidermis of microneedle-treated tissue was demonstrated using light and fluorescent microscopy. Track etched permeation profiles, generated using 'Franz-type' diffusion cell methodology and a model synthetic membrane showed that >50% of a colloidal particle suspension permeated through membrane pores in approximately 2 hours. On the basis of these results, it is probable that microneedle treatment of the skin surface would facilitate the cutaneous delivery of lipid:polycation:pDNA (LPD) gene vectors, and other related vectors, to the viable epidermis. Preliminary gene expression studies confirmed that naked pDNA can be expressed in excised human skin following microneedle disruption of the SC barrier. The presence of a limited number of microchannels, positive for gene expression, indicates that further studies to optimise the microneedle device morphology, its method of application and the pDNA formulation are warranted to facilitate more reproducible cutaneous gene delivery. PMID- 16472097 TI - Topical DermaVir vaccine targeting dendritic cells. AB - DermaVir employs a topical, non-invasive method for vaccine delivery to dendritic cells. The vaccine product contains plasmid DNA as the active ingredient, encoding authentically expressed retroviral genes with appropriate safety modifications. The non-viral delivery system packages the DNA within pathogen like nanoparticles and studies indicate that vaccine antigens are taken up by epidermal Langerhans cells, the precursors of dendritic cells. DermaVir loaded dendritic cells reach the draining lymph node target but not the bloodstream nor indiscriminately other organ systems. Safety data from DermaVir immunized infected macaques indicate improved survival, absence of apparent toxicities other than transient erythema and lack of recombination between the vaccine DNA and the infectious viral DNA integrated in the host genome. DermaVir represents a potential new approach for the treatment of HIV infection to be utilized either in conjunction with antiretroviral therapy or during structured treatment interruption. PMID- 16472098 TI - Gene delivery into human skin in vitro using biphasic lipid vesicles. AB - Topical gene delivery to the skin shows great potential for painless, non invasive administration of novel vaccines and therapeutic agents. The challenge is to develop a pharmaceutically acceptable system that can deliver suitable amounts of plasmid DNA to produce the desired level of response. The purpose of this study was to quantitatively assess DNA delivery by a novel lipid-based biphasic delivery system into the viable layers of excised human skin. Biphasic lipid vesicle formulations, incorporating plasmid DNA were evaluated in vitro in flow-through diffusion cells. Fifty mg DNA formulation containing 10 microg DNA was applied to full-thickness human breast skin for 24 hours. Residual formulation was removed and the skin was washed with PBS, then tape-stripped, followed by DNase treatment to remove surface bound DNA. Skin samples were homogenised and digested overnight with Proteinase K. The resulting supernatant was used as a template for quantitative PCR. Three formulations yielded a significant degree of dermal absorption compared to the controls. Formulation 26 3-2-DNA indicated that approximately 1x10(9) copies of plasmid were absorbed per cm2 skin. Other formulations resulted in 5x10(6) copies/cm2 skin (17C3-1-DNA) and 5x10(8) copies/cm2 skin (26-3-1-DNA). Biphasic vesicles delivered significant quantities of plasmid DNA into the 'viable' layers of human skin in vitro. The successful delivery of this large (approximately 4,400 kDa) charged molecule through intact stratum corneum represents a major advance in transdermal macromolecule delivery. PMID- 16472099 TI - Thermotropic and spectroscopic behavior of skin: relationship with percutaneous permeation enhancement. AB - Stratum corneum (SC) is comprised of lipids, protein and low molecular weight water-soluble components. Changes in these skin micro constituents can be understood by instrumental methods like differential scanning calorimetry (DSC) and attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy. The former provides information about changes in thermotropic behavior of SC lipids and proteins, whereas the latter provides data about alterations at molecular and conformational level. Most of the DSC thermograms of intact mammalian SC show two reversible and two irreversible transitions in the temperature range of 25-125 degrees C. The reversible endotherms are ascribed to lipid melting transitions, whereas the irreversible endotherms are ascribed to protein denaturation. Similarly, the FTIR spectral bands of SC occurring between 2920-2850 cm-1 and between 1650-1550 cm-1 have been suggested to arise from lipid and protein molecular vibrations, respectively. Treatment of skin with solvents or permeation enhancers alters the composition of lipids or their molecular arrangement in the skin microenvironment, which leads to changes in permeability of drug molecules. Furthermore, inhibition of lipid synthesis in epidermis with concomitant decrease in enthalpy of lipid endothermic transitions and reduction in height and area of asymmetric and symmetric C-H stretching peaks have been found to be directly correlated with enhanced permeation of drugs. In addition, method of skin preparation, type of skin, types of enhancer etc. also influence both the nature and intensity of responses recorded in spectrographs and thermograms. Therefore, the modification in spectrographs and thermograms of skin samples treated with various enhancers, vehicles etc. are expected to provide better insight into their mechanism of action on the skin. This review article shall critically evaluate the thermotropic and infrared spectroscopic data of SC/epidermis after various treatments. PMID- 16472100 TI - Polylactide-co-glycolide microparticles with surface adsorbed antigens as vaccine delivery systems. AB - Several groups have shown that vaccine antigens can be encapsulated within polymeric microparticles and can serve as potent antigen delivery systems. We have recently shown that an alternative approach involving charged polylactide co glycolide (PLG) microparticles with surface adsorbed antigen(s) can also be used to deliver antigen into antigen presenting cell (APC). We have described the preparation of cationic and anionic PLG microparticles which have been used to adsorb a variety of agents, which include plasmid DNA, recombinant proteins and adjuvant active oligonucleotides. These PLG microparticles were prepared using a w/o/w solvent evaporation process in the presence of the anionic surfactants, including DSS (dioctyl sodium sulfosuccinate) or cationic surfactants, including CTAB (hexadecyl trimethyl ammonium bromide). Antigen binding to the charged PLG microparticles was influenced by several factors including electrostatic and hydrophobic interactions. These microparticle based formulations resulted in the induction of significantly enhanced immune responses in comparison to alum. The surface adsorbed microparticle formulation offers an alternative and novel way of delivering antigens in a vaccine formulation. PMID- 16472101 TI - Particulate and vesicular drug carriers in the management of tuberculosis. AB - Although oral drug therapy for tuberculosis exists and is widely followed, its major drawbacks are lack of patient compliance and development of adverse effects like hepatotoxicity on long term use. Absence of new therapeutic agents and the above mentioned demerits have led to search for alternative methods for delivery of antitubercular agents. Colloidal drug carriers, a popularly utilized delivery system has been deeply explored for the cause. The article discusses the advances in the management of tuberculosis by the use of particulate and vesicular drug carriers by parenteral, inhalational and oral routes. Use of this delivery strategy has led to massive reduction in the dosage resulting in toxicity alleviation. As a number of studies have already been undertaken in experimental models, it will be a promising tool in the prevention of relapse and successful treatment of tuberculosis in patients. PMID- 16472102 TI - Induction of cytochrome P450 3A4 and P-glycoprotein by the isoxazolyl-penicillin antibiotic flucloxacillin. AB - Clinical findings indicate that co-administration of the isoxazolyl-penicillin flucloxacillin with cyclosporine may reduce the plasma concentrations of cyclosporine. We have explored in the present study if induction of cytochrome P450 3A4 or P-glycoprotein may offer a mechanistic explanation of the observed effects. Flucloxacillin is neither an inhibitor nor a substrate of drug metabolizing cytochrome P450 isoenzymes (CYP3A4, 1A2, 2C9, 2C19 and 2D6) or P glycoprotein as shown by an in vitro assay for CYP inhibition, a fluorescent indicator assay for P-glycoprotein inhibition and a functional P-glycoprotein ATPase assay. However, incubation of human LS 180 colorectal adenocarcinoma cells with flucloxacillin led to a dose-dependent induction of MDR1 as well as of CYP3A4 mRNA, which was also confirmed in primary human hepatocytes. At high concentrations, flucloxacillin activated the human Pregnane-X-Receptor, PXR, a ligand-dependent transcription factor that is the target of many drugs that induce CYP3A4, with consequences for the metabolism of other drugs. Liver microsomes from control rats or rats, which received for 3 consecutive days 100 mg/kg of oral flucloxacillin, were used to study the metabolism and metabolite pattern of midazolam, a model substrate of CYP 3A4. There was a trend towards a higher intrinsic microsomal clearance of midazolam using microsomes from flucloxacillin treated rats. In addition, there was a significant increase in the formation of the principal midazolam metabolites 1-hydroxy midazolam, 4-hydroxy midazolam and 1,4-dihydroxy midazolam as compared to controls. These findings indicate that flucloxacillin has the potential to induce expression of both CYP3A4 as well as P-glycoprotein, most likely through activation of the nuclear hormone receptor PXR. This would offer an explanation for the observed clinical drug-drug interactions between the antibiotic and cyclosporine. PMID- 16472103 TI - Fluoxetine metabolism and pharmacological interactions: the role of cytochrome p450. AB - A review with 103 references. Fluoxetine is the parent drug of the SSRI (selective serotonin reuptake inhibitor) antidepressant class, and is still one of the most highly used drugs of this class world-wide. Fluoxetine now has largely (albeit not completely) substituted older and less safe drugs such as tricyclic antidepressants. Different cytochrome P450 isoforms are involved in the metabolism of fluoxetine, however, the main active metabolite, norfluoxetine, is produced by the CYP2D6 action in the human liver. In this paper, the main metabolic characteristics of fluoxetine will be reviewed, with particular attention paid to the role of cytochrome isozymes. The pharmacological interactions of the drug will be overviewed, especially those concerning other drugs used in psychiatric clinics, such as antipsychotics and antidepressants and the relationships between pharmacological interactions and cytochrome activity will be discussed. Recently, much attention has been drawn to the therapeutic drug monitoring (TDM) of fluoxetine, and in particular to the analysis of fluoxetine enantiomers for which enantiomeric separations and enantioselective metabolism will also briefly be mentioned. PMID- 16472104 TI - Prediction of drug-drug interactions for AUCoral of high clearance drug from in vitro data: utilization of a microtiter plate assay and a dispersion model. AB - The purpose of this study was to propose a new method to predict in vivo drug drug interactions (DDIs) for a high clearance drug from in vitro data. As the high clearance drug, NE-100 (N, N-dipropyl-2-[4-methoxy-3-(2 phenylethoxy)phenyl]ethylamine monohydrochloride) was used. First, approach based on I(u)/K(i) value was used for the prediction of DDIs between NE-100 and concomitant drugs. When the K(i) values (K(i-cal)) obtained from the microtiter plate (MTP) assay and the reported K(i) values (K(i-rep)) for these drugs were used to predict increases at levels of NE-100 AUC(oral) (AUC(oral) ratio), the AUC(oral) ratios from the I(u)/K(i-cal) correlated with those from the I(u)/K(i rep). This result suggests that the K(i-cal) from the MTP assay can be used for prediction of DDIs instead of the K(i-rep) value. Second, a new approach combining the inhibition rate (R) calculated from the MTP assay and two physiological models was used to predict DDIs. When the AUC(oral) ratios of NE 100 by various drugs were predicted using the R value and the well-stirred model, the ratios were similar to those predicted using the I(u)/K(i). However, after co administration of drugs such as quinidine, propafenone and thioridazine (potent inhibitors of CYP2D6), the NE-100 AUC(oral) ratios predicted from the dispersion model was much greater than those from well-stirred model. This result shows that application of the dispersion model to the prediction method using the R value might sensitively and precisely predict the increased levels of AUC(oral) by DDIs for high clearance drug, compared with the prediction method using I(u)/K(i) value. PMID- 16472105 TI - Predicting the pharmacokinetics of acyl glucuronides and their parent compounds in disease states. AB - Acyl glucuronides are potentially reactive intermediates, which not only undergo hydrolysis and intramolecular acyl migration, but also bind irreversibly to plasma protein in vitro and in vivo. To evaluate the impact of renal failure, liver dysfunction and other disease states on the pharmacokinetics of acyl glucuronides and their parent compounds, a pharmacokinetic model has been established. The model has been successfully utilized to predict the pharmacokinetics of six compounds, diflunisal (DF), valproic acid (VPA), zomepirac (Z), suprofen (S), R-etodolac (R-ET), S-etodolac (S-ET), and their acyl glucuronides in various simulated disease states in experimental animals. Modeling studies revealed that altering the metabolic pathways of these compounds had significant impact on exposure and clearance of acyl glucuoninde. The simulation results also indicated that disease states that affect irreversible metabolic pathways other than glucuronidation may have major impacts on the apparent plasma clearance of the parent compound or exposure to the reactive acyl glucuronide as well. The study concluded that the model is sufficiently robust and applicable for pharmacokinetic studies of acyl glucuronides and their parent compounds in various disease states that may modulate drug clearance. The model is also applicable to understanding the complex disposition of other drugs subject to conjugation, especially those that can be reversible and undergo enterohepatic recycling, such as sulfation and glycine conjugation. PMID- 16472107 TI - The role of blood-brain barrier studies in the pharmaceutical industry. AB - The blood-brain barrier (BBB) remains one of the greatest challenges for the discovery and development of treatments for CNS disorders, which to this day remains one of the riskiest disease areas in terms of clinical success rates. Although the BBB is currently seen predominantly as a permeability obstacle for CNS drug delivery, it is becoming increasingly clear that the BBB has many more implications for the pharmaceutical industry impacting on CNS pharmacology and pathology, CNS pharmacokinetics and pharmacodynamics, and adverse CNS effects, to name but a few areas. The present review does not intend to summarize the activities in the field of BBB research per se, which has been accomplished by a number of excellent recent reviews, but instead to provide an overview of the role of BBB studies from a pharmaceutical industry perspective. This review will elaborate on the specific needs in terms of BBB-related issues across the different drug discovery and development phases, i.e. target identification and validation, lead generation and optimization, candidate selection and profiling, preclinical development and clinical studies. The specific approaches taken will be discussed in terms of specific requirements, questions to be asked, feasibility, interpretability, and impact. It becomes clear that few of the existing BBB models fully meet the requirements of the industrialized drug discovery process, highlighting the need for an array of new or modified tools and approaches that are more effective in helping make decisions which are more specifically tailored to the various stages of the lengthy process from target to the clinic. In looking at the numerous ongoing activities in the area of BBB research from the drug discovery and development point of view, an attempt has been made to place a stronger emphasis on the applicability of particular techniques and approaches, to identify gaps and areas for future activities. In order to materialize the considerable knowledge gained in recent years, the review is intended to foster an increased awareness of the need to better integrate basic academic research with the specific requirements of the pharmaceutical industry for the search of effective and safe new CNS medicines. PMID- 16472106 TI - Preclinical pharmacokinetics: an approach towards safer and efficacious drugs. AB - Lack of efficacy and toxicity are considered to be major reasons for drug failures and pharmacokinetics governs them to a large extent. Compound with favorable pharmacokinetics is more likely to be efficacious and safe. Therefore, the preclinical pharmacokinetic evaluation should be comprehensive enough to ensure that compounds do not fail in the clinic. Preclinical ADME screening facilitates early elimination of weak candidates and directs the entire focus of the drug development program towards fewer potential lead candidates. Hence, it is mandatory that the pre-clinical candidates are subjected to as many possible reality checks. Reliance on in-vitro tests should be minimized because they do not represent the real physiological environment but rather slow down the pace of a drug discovery program. Compounds can be straight away subjected to in-vivo high throughput screens such as cassette dosing, cassette analysis or rapid rat screen etc. Candidates with the desired in-vivo pharmacokinetic profile may be further profiled in-vitro, using assays such as metabolic stability, reaction phenotyping, CYP-450 inhibition and induction, plasma protein binding etc. in human microsomes, human recombinant CYP-450 enzymes and human plasma. This also provides an early indication of whether the compound which worked in animals would work in human as well. In-vitro metabolic stability profile is a qualitative as well as quantitative comparison of metabolism of a compound in human and animal models. It helps in identifying the right model for toxicity studies. Extensive metabolism is generally considered a liability as it limits the systemic exposure and shortens the half-life of a compound. Several strategies such as reduction of lipophilicity, modification and / or blocking of metabolically soft spots and use of enzyme inhibitors; have been developed to combat metabolism. In spite of several concerns, the fact that active metabolites of several marketed drugs have been developed as drugs with better efficacy, safety and pharmacokinetics profile; cannot be denied. Therefore, instead of considering metabolic instability a liability it can be exploited as a tool for discovering better drugs. It is equally important to identify the metabolic pathways of the drug candidates by conducting in-vitro CYP450 reaction phenotyping assays. The identification of drug metabolizing enzymes involved in the major metabolic pathways of a compound helps in predicting the probable drug drug interactions in human. Compounds with more than one metabolic pathway have less likelihood of clinically significant drug interactions. In-vitro CYP450 inhibition and induction screens are used to evaluate the potential of compound towards drug - drug interactions and the most prone candidates may either be discarded or taken ahead with a caution. It is known that only unbound drug is pharmacologically active and therefore the assessment of bound fraction by the estimation of plasma protein binding of a compound is another important parameter to be explored in-vitro. In addition to the process of 'weeding out' weak candidates early in the drug discovery process, it is equally important to identify the probable causes of poor ADME exhibited by some compounds as this information is useful to medicinal chemists for improving upon backbones that exhibit un favorable pharmacokinetic profile. Toxicity study is the foundation of an INDA (Investigational new drug application) and therefore, the final selection of a compound can be performed only after proper toxicological evaluation in animal models. Toxicokinetics forms an integral part of toxicity study and is used to assess the exposure of candidates in toxicity models and correlate the drug levels in blood and various tissues with the toxicological findings. Although in-vivo screening of compounds in animal models and in-vitro assays in human recombinant CYP-450 enzymes help in drug candidate selection, both approaches have their own limitations. There is no certainty that the selected candidates will exhibit the desired target PK profile in human and real human PK remains suspense until the compound enters Phase-1 clinical trial. The recognition of human micro dosing, (HMD) by medicines and healthcare products regulatory agency (MHRA) and European agency for evaluation of medicinal products [EMEA] is a stepping stone in the direction of obtaining human PK data early in the preclinical stage. This would gradually shift the focus of early drug development away from animal studies directly towards safe and ethical studies in human yielding more relevant and reliable pharmacokinetic data. HMD would provide an answer to the growing public demand for a reduction in the use of animals for pharmaceutical development. PMID- 16472108 TI - Recent advances in molecular modeling and medicinal chemistry aspects of phospho glycoprotein. AB - Phospho-glycoprotein (P-gp) is an efflux transporter expressed in many organs (ex: kidney, lung, liver and spleen) and in hormone producing or responsive tissues (ex: adrenal cortex, testis and placenta). It is involved in many important physiological functions. Among them the major one is extrusion of xenobiotics in order to detoxify the cells. This property of P-gp is associated with multidrug resistance (MDR) for many pathological conditions. While the experimental determination of three-dimensional structure is not yet successful, the transmembrane (TM) 5, 6, 11 and 12 are sensitive to mutations and contain substrate binding sites. Designing of potential and selective inhibitors of P-gp is still hampered by a lack of information upon the three dimensional structure of P-gp. The design of P-gp inhibitors was traditionally driven by quantitative structure activity relationship studies, which is complicated by factors such as different types of assays, multiple drug binding sites and diverse chemical structures. Clearly a conclusive and predictive SAR does not seem to be practical, despite progress in the last few years towards more specific SAR suggesting well defined structural features responsible for activity. Advances made recently in solving the crystal structure of prokaryotic ATP binding cassette proteins (ABC) transporters, Ec-MsbA, Vc-MsbA and BtuCD yielded suitable templates for construction of homology models of P-gp. Few molecular dynamics (MD) simulations aimed at elucidating the functional dynamics of ABC transporters have provided useful insights to their mechanism and structure. The present review aims at the general overview of importance, expression, structure, organization and drug binding sites of P-gp. This review also highlights recent developments in the homology modeling, molecular dynamics simulations of P-gp and progress in QSAR, pharmacophore modeling of P-gp modulators. PMID- 16472109 TI - Emerging roles of the unfolded protein response signaling in physiology and disease. PMID- 16472110 TI - Divergent roles of IRE1alpha and PERK in the unfolded protein response. AB - The endoplasmic reticulum (ER) provides unique machinery for the folding and posttranslational modification of many secretory and transmembrane proteins in eukaryotic cells. The unfolded protein response (UPR) is a signal transduction network from the ER to the nucleus activated when the folding demand imposed by nascent, unfolded polypeptide chains exceeds the capacity of the ER protein folding machinery. In all eukaryotes the UPR maintains the physiological balance between folding demand and capacity of the ER by regulating adaptive responses to this stress situation. These include an increase in the folding capacity of the ER through induction of ER resident molecular chaperones and protein foldases, and a decrease in the folding demand on the ER by upregulation of ER associated degradation (ERAD), attenuation of general translation in metazoans, and stimulation of ER synthesis to dilute the unfolded protein load. In higher eukaryotes the UPR gained control over inflammatory and immune responses by controlling the activity of the transcription factor NF-kappaB to combat viral infections associated with an increased synthesis of viral glycoproteins. Similarly, in multicellular organisms apoptotic programs are controlled by the UPR to eliminate cells whose folding problems in the ER cannot be resolved by coordinated regulation of adaptive, inflammatory, and immune responses. In this review we will summarize our current understanding of signal transduction mechanisms involved in the mammalian UPR, and discuss examples to highlight the regulation of adaptive, inflammatory, immune, and apoptotic responses by the UPR. PMID- 16472111 TI - Stressing out the ER: a role of the unfolded protein response in prion-related disorders. AB - Transmissible Spongiform Encephalopathies are fatal and infectious neurodegenerative diseases characterized by extensive neuronal apoptosis and the accumulation of an abnormally folded form of the cellular prion protein (PrP), denoted PrP(SC). Compelling evidence suggests the involvement of several signaling pathways in prion pathogenesis, including proteasome dysfunction, alterations in the protein maturation pathways and the unfolded protein response. Recent reports indicate that endoplasmic reticulum stress due to the PrP misfolding may be a critical factor mediating neuronal dysfunction in prion diseases. These findings have applications for developing novel strategies for treatment and early diagnosis of transmissible spongiform encephalopathies and other neurodegenerative diseases. PMID- 16472112 TI - Stress induction of GRP78/BiP and its role in cancer. AB - GRP78, also referred to as BiP, is a central regulator of endoplasmic reticulum (ER) function due to its roles in protein folding and assembly, targeting misfolded protein for degradation, ER Ca(2+)-binding and controlling the activation of trans-membrane ER stress sensors. Further, due to its anti apoptotic property, stress induction of GRP78 represents an important pro survival component of the unfolded protein response. GRP78 is induced in a wide variety of cancer cells and cancer biopsy tissues. Recent progress, utilizing overexpression and siRNA approaches, establishes that GRP78 contributes to tumor growth and confers drug resistance to cancer cells. The discovery of GRP78 expression on the cell surface of cancer cells further leads to the development of new therapeutic approaches targeted against cancer, in particular, hypoxic tumors where GRP78 is highly induced. Progress has also been made in understanding how Grp78 is induced by ER stress. The identification of the transcription factors interacting with the ER stress response element leads to the discovery of multiple pathways whereby mammalian cells can sense ER stress and trigger the transcription of Grp78. In addition, advances have been made in understanding how Grp78 expression is regulated in the context of chromatin modification. This review summarizes the transcriptional regulation of Grp78, the molecular basis for the cytoprotective function of GRP78 and its role in cancer progression, drug resistance and potential future cancer therapy. PMID- 16472113 TI - ER stress, hypoxia tolerance and tumor progression. AB - The development of chronic and fluctuating hypoxic regions in tumors has profound consequences for malignant progression, response to therapy and overall patient survival. Understanding the events involved in hypoxia tolerance will offer new opportunities for antitumor modalities. A universal response of tumor cells to hypoxia is a rapid and substantial decrease in the rates of macromolecular synthesis. Hypoxia induces phosphorylation of the translation initiation factor eIF2alpha on Ser51 via activation of the endoplasmic reticulum (ER) resident kinase PERK and that this modification is required for the rapid downregulation of global protein synthesis by this hypoxic stress. PERK-dependent phosphorylation of eIF2alpha is one component of the Unfolded Protein Response (UPR), a coordinated program that promotes cell survival under conditions of ER stress. Inactivation of PERK or eIF2alpha phosphorylation impairs cell survival under hypoxia, and transformed cells with inactivating PERK or eIF2alpha mutations form tumors in nude mice that are slower growing, and have higher levels of apoptosis in hypoxic areas compared to tumors with an intact UPR. Expression of the transcription factor ATF4, a downstream effector of eIF2alpha phosphorylation, is also upregulated by hypoxia in vitro and in human tumors and increases hypoxia tolerance. A second UPR pathway mediated by activation of IRE1 and its downstream target XBP1 is also required for hypoxia tolerance in vitro and for tumor growth. These results reveal a critical role for UPR activation for tumor cell resistance to hypoxia and tumor growth promotion and suggest that the UPR may be an attractive target for anti-tumor modalities. PMID- 16472114 TI - Endoplasmic reticulum stress-induced apoptosis and auto-immunity in diabetes. AB - Increasing evidence suggests that stress signaling pathways emanating from the endoplasmic reticulum (ER) are important to the pathogenesis of both type 1 and type 2 diabetes. Recent observations indicate that ER stress signaling participates in maintaining the ER homeostasis of pancreatic beta-cells. Either a high level of ER stress or defective ER stress signaling in beta-cells may cause an imbalance in ER homeostasis and lead to beta-cell apoptosis and autoimmune response. In addition, it has been suggested that ER stress attributes to insulin resistance in patients with type 2 diabetes. It is necessary to study the relationship between ER stress and diabetes in order to develop new therapeutic approaches to diabetes based on drugs that block the ER stress-mediated cell death pathway and insulin resistance. PMID- 16472115 TI - ER stress and UPR in familial amyotrophic lateral sclerosis. AB - The primary mechanism by which mutations in Cu, Zn-superoxide dismutase (SOD1) contribute to progressive motor neuron loss in familial amyotrophic lateral sclerosis (FALS) remains unknown. Misfolded protein aggregates, ubiquitin proteasome system impairment and neuronal apoptosis mediated by death receptor or mitochondrial-dependent pathways are implicated in mutant SOD1-induced toxicity. Recent evidence from cellular and transgenic rodent models of FALS proposes activation of a third apoptotic pathway linked to sustained endoplasmic reticulum (ER) stress. Here, we review the emerging role of ER stress and the unfolded protein response (UPR) in the pathogenesis of mutant SOD1-linked FALS. The UPR observed in FALS rodents is described which encompasses induction of key ER resident chaperones during presymptomatic disease, leading to activation of stress transducers and pro-apoptotic molecules by late stage disease. Importantly, mutant SOD1 co-aggregates with UPR components and recruits to the ER, suggesting a direct adverse effect on ER function. By contrast, the opposing neuroprotective effects of wild-type SOD1 overexpression on UPR signalling are also highlighted. In addition, the potential impact of neuronal Golgi apparatus (GA) fragmentation and subsequent disturbances in intracellular protein trafficking on motor neuron survival in FALS is also discussed. We propose that ER stress and UPR may be coupled to GA dysfunction in mutant SOD1-mediated toxicity, promoting ER-initiated cell death signalling in FALS. PMID- 16472116 TI - The ASK1-MAP kinase signaling in ER stress and neurodegenerative diseases. AB - Accumulation of unfolded and/or misfolded proteins in the endoplasmic reticulum (ER) lumen induces ER stress. ER stress triggers the unfolded protein response (UPR), which includes the attenuation of general protein synthesis and the transcriptional activation of the genes encoding ER-resident chaperones and molecules involved in the ER-associated degradation (ERAD). The UPR coordinately reduces ER stress by restoration of the protein-folding capacity of the ER. However, severe and/or prolonged ER stress eventually leads cells to apoptosis. Several lines of evidence suggest that ER stress-induced apoptosis plays critical roles in the pathogenesis of neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, polyglutamine (polyQ) diseases and amyotrophic lateral sclerosis (ALS). Apoptosis signal-regulating kinase 1 (ASK1), a member of the MAPKKK family that constitutes the JNK and p38 MAP kinase (MAPK) cascades, is activated by physiological and cytotoxic stresses and induces various stress responses including apoptosis. Recent studies have shown that the ASK1-MAPK cascades are involved in ER stress-induced apoptosis and in the neuronal cell death in some model systems of neurodegenerative diseases. This review highlights the current understanding of regulatory mechanisms of ASK1 with a special focus on the ER stress-dependent and -independent neuronal cell death in the context of neurodegenerative diseases. PMID- 16472117 TI - The control of endoplasmic reticulum-initiated apoptosis by the BCL-2 family of proteins. AB - Irreversible perturbations in the homeostasis of the endoplasmic reticulum (ER) are thought to lead to apoptosis and cell loss in a number of important human diseases, including Alzheimer disease, Parkinson disease, and type 2 diabetes. However, the exact mechanisms that lead from ER stress to cell death remain incompletely understood. Recent work has shown that the BCL-2 family of proteins plays a central role in regulating this form of cell death, both locally at the ER and from a distance at the mitochondrial membrane. PMID- 16472118 TI - Conformational diseases and ER stress-mediated cell death: apoptotic cell death and autophagic cell death. AB - The expanded polyglutamine (polyQ) tracts observed in autosomal dominant neurodegenerative disorders have the tendency to form intracellular aggregates, thus enhancing apoptotic cell death and the formation of autophagic vesicles. PolyQ accumulation inhibits the ER-associated degradation system (ERAD) resulting in reduced retrotranslocation from the ER and increased accumulation of misfolded proteins in the lumen of ER. Autophagy is an early cellular defense mechanism associated with ER stress, but prolonged ER stress may induce autophagic cell death, with destruction of cellular components and apoptotic cell death. Endoplasmic reticulum (ER) stress may be the key signal for both of these events. PMID- 16472119 TI - The role of the endoplasmic reticulum in the accumulation of beta-amyloid peptide in Alzheimer's disease. AB - Increased cerebral levels of Abeta(42) peptide, either as soluble or aggregated forms, are suggested to play a key role in the pathogenesis of Alzheimer's disease (AD). The identification of genetic defects in presenilins and beta amyloid precursor protein (beta-APP) has led to the development of cellular and animal models that have helped in understanding aspects of the pathophysiology of the inherited early onset forms of AD. However, the majority of AD cases are sporadic with no clear or defined genetic basis. While genetic mutations are responsible for the accumulation of Abeta in early onset AD, the causative factors for accumulation of Abeta in the late onset AD forms are not known. This raises the possibility that Abeta accumulation in the absence of genetic mutations might result from abnormalities that indirectly affect Abeta production or its clearance. Currently, there is no consensus as to what are the mechanisms by which Abeta accumulates or as to which mechanisms underlie Abeta-induced neuronal death in AD. In this review, I will first describe the physiological role of endoplasmic reticulum in the cell and review some of the data supporting dysfunction of the endoplasmic reticulum as an early event leading to Abeta accumulation in familial AD. I will also discuss the possible role of oxidative stress and other factors as contributors in Abeta accumulation by reducing the clearance of Abeta from the endoplasmic reticulum. Finally, I will summarize data that show the endoplasmic reticulum stress as a mechanism underlying exogenous Abeta neurotoxicity. PMID- 16472121 TI - Accelerated delimitation of the infarct zone by capillary-derived nestin-positive cells in aged rats. AB - An important cellular event associated with reduced structural and functional recovery after stroke in aged animals is the early formation of a scar in the infarcted region that impairs neural recovery and repair. Despite the detrimental impact of infarct scar formation, the brain regions and cell types that supply the components of the scar are not well characterized. We hypothesized that premature cerebral scar formation in aged animals is associated with an altered cellular response to cerebral ischemia. Focal cerebral ischemia was produced by reversible occlusion of the right middle cerebral artery in 3 month- and 20 month old male Sprague Dawley rats. After 3, 7, 14, and 28 days, brain tissue was subjected to real-time reverse-transcriptase-PCR (RT-PCR) and immunostaining for 1) a cellular proliferation marker (BrdU); 2) a neuroepithelial marker (nestin); 3) an astrocytic marker (glial fibrillary acidic protein [GFAP]); 4) a neuronal marker, doublecortin; and 5) a basal lamina marker (laminin), and analyzed using 3D-reconstruction of confocal images. In this model the infarct was localized primarily in the parietal cortex. By RT-PCR there was a robust increase in nestin mRNA transcripts shortly after stroke, and this increase was particularly intense in aged rats. Accordantly, we found in aged rats a rapid delimitation of the infarct area by nestin-positive cells and an early incorporation of these cells into the glial scar. The capillaries of the corpus callosum were the major source of proliferating, nestin-positive cells, many of which were also immunoreactive for doublecortin, although a smaller population of nestin cells were associated with the ventricular walls. Despite the proliferation of nestin cells, they did not make a significant contribution to neurogenesis in the infarcted cortex, possibly because the corpus callosum impedes the migration of subventricular zone derived nestin-positive cells into the lesioned area. We conclude that: (i) the aged brain has the capability to mount a cytoproliferative response to injury, but the timing of the cellular and genetic reaction to cerebral insult is accelerated in aged animals; (ii) the proliferating cells contribute to the formation of the glial scar, but few of the cells appear to become neurons; and (iii) the vasculature plays a hitherto unrecognized role as a source of proliferating cells after stroke. Because capillary-derived cells help to form the glial scar, elucidating the molecular basis of this phenomenon and its acceleration in the aging brain could yield novel approaches to enhancing neurorestoration in the elderly. PMID- 16472122 TI - Cerebral angiogenesis and expression of angiogenic factors in aging rats after exercise. AB - The effect that exercise has on angiogenesis in the aging rat is unknown. We initiated this study with the intent to determine if exercise could induce angiogenesis in aging rats, as well as in adult rats reported previously. The markers we used to determine our endpoint were vascular endothelial growth factor (VEGF) and angiopoietin 1 and 2, as well as vascular density. Aged (22 month old) female Fisher 344 rats (n=16) were exercised on a treadmill 30 minutes each day for 3 weeks, or housed as non-exercised controls for the same duration. At the end of the exercise protocol, a significant (p<0.01) increase in the density of microvessels was found within the cerebral vasculature of the rats. Exercise was also associated with a significantly (p<0.01) increased mRNA expression of angiopoietin 1 and 2 in the aged cohort of rats. A mild but significant (p<0.01) increase in the four isoforms of VEGF mRNA (120, 144, 164, 188) were observed, with VEGF120 and VEGF144 being more markedly up-regulated than the other two. VEGF protein expression was also significantly (p<0.01) increased. This study demonstrates that angiogenesis can be induced in aging rats via exercise. The induced angiogenesis was associated with overexpression of angiogenic factors. These results support the hypothesis that an angiogenic response to chronic physical exercise is maintained with aging. PMID- 16472124 TI - Lovastatin reduces apoptosis and downregulates the CD40 expression induced by TNF alpha in cerebral vascular endothelial cells. AB - Inflammation may be one of the independent risk factors contributing to many neurological diseases. Moreover, there is an emerging body of data indicating that statins may have neuroprotective action. Recent studies suggest that CD40 CD40 ligand (CD40L) system is proven to be an important mediator of several auto immune and chronic inflammation diseases. To address whether lovastatin produces neuroprotection as a potential novel anti-inflammatory pathway through the inhibition of CD40 expression, we examined the possible effects of lovastatin on expression of CD40, apoptosis, level of nitric oxide (NO) and nitric oxide synthase (NOS) activity induced by tumor necrosis factor alpha (TNF-alpha) in the cerebral vascular endothelial cells (CVECs) involved in cerebrovascular diseases. Preincubation with lovastatin (10(-7), 10(-6) and 10(-5) mol/l) for 24 hours (h) protected CVECs from TNF-alpha-induced decrease of cellular viability. Further, lovastatin inhibited the TNF-alpha-induced increases of NO level, NOS activity, apoptotic cells and CD40 expression in a dose-dependent manner, and anti-CD40 antibody also inhibited the cellular apoptosis induced by TNF-alpha. In conclusion, our data provide evidence to support a direct pro-inflammatory effect of CD40-CD40L signaling pathway in CVECs, and lovastatin possesses an anti inflammatory effect independent of its lipid-lowering action involved in the cerebrovascular diseases. PMID- 16472125 TI - High glucose-mediated imbalance of nitric oxide synthase and dimethylarginine dimethylaminohydrolase expression in endothelial cells. AB - The mechanisms involved in endothelial dysfunction are multifactorial. A correlation between oxidative stress and derangements of nitric oxide synthase (NOS) pathways in altered endothelial homeostasis has been most studied and demonstrated in different pathophysiological conditions. NOS activities are regulated by endogenous inhibitors such as asymmetric dimethyl-L-arginine (ADMA) that is metabolized by dimethylarginine dimethylaminohydrolase (DDAH). Since recent data demonstrated that some endothelial dysfunction may be related to reduced expression and/or activity of DDAH, the aim of the present research was to investigate the expression of DDAH-2 and NOS isoforms in high glucose-mediated oxidative stress. Endothelial cells were incubated with normal (7 mM) and high concentrations (33 mM) of D-glucose for 5 days; mannose (26 mM) plus D-glucose (7 mM) was used as osmotic control. Data obtained in the present study show that the exposure for 5 days to high glucose increases oxidative stress, reduces DDAH-2 and eNOS expression and increases iNOS expression. These results indicate that DDAH-2 and iNOS/eNOS dysregulation may play a key role in high glucose-mediated oxidative stress, suggesting that selective modulation of DDAH isoforms may result in selective inhibition/activation of NOS isoforms, thereby providing a novel strategy of approach in vascular complications of several pathologies. PMID- 16472123 TI - Attempted cell cycle induction in post-mitotic neurons occurs in early and late apoptotic programs through Rb, E2F1, and caspase 3. AB - Either the absence or dysfunction of a number of critical pathways, such as those that involve the nuclear retinoblastoma protein (Rb) and the transcription factor E2F1, may account for the aberrant induction of the cell cycle in post-mitotic neurons that can be responsible for oxidative stress-induced apoptotic cellular destruction. Yet, it is unclear whether early programs of apoptotic injury that involve membrane phosphatidylserine (PS) exposure and calreticulin expression as well as later phases of apoptotic injury with nuclear DNA injury require the critical modulation of Rb and E2F1. We demonstrate that both the post translational of phosphorylation of Rb to prevent E2F1 transcription as well as the protein integrity of Rb are closely aligned with the modulation of cell cycle induction in post mitotic neurons during oxidative stress. More importantly, we illustrate that both the initial onset of apoptosis with either membrane PS exposure or calreticulin analysis as well as the more terminal phases of apoptosis that involve nuclear DNA degradation proceed concurrently in the same neuronal cells with cell cycle induction. Progression of attempted cell cycle induction is closely associated with the phosphorylation of Rb, its inability to bind to E2F1, and the degradation of the Rb protein. Inhibition of Rb phosphorylation using cyclin dependent kinase inhibitors maintains the integrity of the E2F1/Rb complex and is neuroprotective during free radical exposure. Furthermore, maintenance of the integrity of the Rb protein is specifically dependent upon caspase 3-like activity, since caspase 3 can cleave Rb during free radical activity and this degradation of Rb can be blocked during the inhibition of caspase 3 activity. Our studies not only highlight the critical role of attempted cell cycle induction during oxidative stress-induced neuronal apoptotic injury, but also bring to light the significant impact of the Rb and E2F1 pathways upon early apoptotic programs that can directly influence both intrinsic cell survival as well as extrinsic inflammatory cell activation. PMID- 16472126 TI - Structure and function of myelinated nerve fibers in the rabbit eye following ischemia/reperfusion injury. AB - The rabbit eye presents a valuable model to study the effects of vascular occlusion on the function and structure of myelinated nerve fibers. The rabbit eye has a band of myelinated nerve fibers within the intraocular compartment that are supplied by a narrow band of retinal vasculature. These vessels were transiently occluded ( approximately 8 hours) using laser photocoagulation and the transmission of electrical signals along the nerve fibers was assessed by recording the visual evoked response (VER). Morphological damage was assessed by histological techniques. The ischemic insult produced no permanent change in retinal function as assessed by electroretinography, but the VER was suppressed, indicating failure of nerve fiber transmission. Histologically, the visible damage to the region supported by the retinal vasculature worsened following reperfusion, showing evidence of demyelination and necrosis followed by macrophage responses and gliosis. This rabbit model of ischemia/reperfusion of the retinal vasculature offers a rare opportunity to reliably study the response of myelinated nerve fibers to ischemia/reperfusion insults and has demonstrated the susceptibility of myelinated nerve fibers to such insults. PMID- 16472127 TI - Stroke-induced neurogenesis: physiopathology and mechanisms. AB - Long-term disabilities are the main outcome of cerebral strokes, though some of the deficits show receding signs in the weeks and months following the "brain attack". Studies show that neurogenesis is induced in the hippocampus and subventricular zone (SVZ) in animal models of ischemia, and that new neurons are generated at the sites of degeneration, where they replace some of the lost nerve cells. The enhanced neurogenesis suggests the involvement of the hippocampus and SVZ in the physiopathology of cerebral strokes, and the generation of new neuronal cells at the sites of degeneration suggests that the central nervous system (CNS) may attempt to repair itself. In this manuscript, we will review the studies on adult neurogenesis in cerebral strokes, discuss the contribution of adult neurogenesis to the physiopathology of strokes, and its underlying mechanisms. PMID- 16472128 TI - Role of advanced glycation end products (AGEs) in thrombogenic abnormalities in diabetes. AB - Accelerated atherosclerosis and microvascular complications are perhaps the leading cause of coronary heart disease, blindness and renal failure, which could account for disabilities and high mortality rates in patients with diabetes. Several thrombogenic abnormalities have been shown to play a central role in the pathogenesis of these devastating complications. However, the molecular mechanism for thrombogenic diathesis in diabetes is not fully elucidated. A recent clinical study, the Diabetes Control and Complications Trial-Epidemiology of Diabetes Interventions and Complications (DCCT-EDIC) Research, has revealed that the reduction in the risk of progressive retinopathy and nephropathy resulting from intensive therapy in patients with type 1 diabetes persist for at least several years, despite increasing hyperglycemia. Further, intensive therapy during the DCCT resulted in decreased progression of carotid intima-media thickness six years after the end of the trial as well. These clinical studies strongly suggest that so-called 'hyperglycemic memory' causes chronic abnormalities in diabetic vessels that are not easily reversed, even by subsequent, relatively good control of blood glucose. Among various biochemical pathways activated under diabetes, the process of formation and accumulation of advanced glycation end products (AGEs) and their mode of action are most compatible with the theory 'hyperglycemic memory'. In this review, we discuss the role of AGEs in thrombogenic abnormalities in diabetes. PMID- 16472129 TI - Identification of genes for a complex trait: examples from hypertension. AB - Essential hypertension (EH) affects approximately 20% of the adult population, and has a multifactorial origin arising from an interaction between susceptibility genes and environmental factors. Several strategies and methods have been used to identify hypertension susceptibility genes. This review is thought to highlight current strategies for a better understanding of their limitations and strengths in a complex trait like EH. Linkage analysis is less effective at identifying common variants with modest effects typical for complex traits, and has therefore proved to be largely unsuccessful in EH. No candidate gene was assessed by a human linkage study so far. Possible redesigns of the linkage approach for complex diseases may include larger sample sizes and dense marker maps. Genetic association studies may be an effective approach to the problems posed by complex traits. With the explosion of genotyping technologies, genome-wide association studies have become feasible, and small-scale association studies have become plentiful. The different types of association studies are reviewed and issues that are important to consider when interpreting association studies of complex traits are discussed. Properly defined phenotypes, large enough sample cohorts to achieve sufficient statistical power, carefully matched samples to avoid population stratification are all integral parts of a high quality association study. Multiple testing often results in false-positive results by chance, and inconclusive results may arise from ignoring linkage disequilibrium of the tested polymorphism, an effect avoidable by haplotype analysis. A new evolutionary development of the candidate gene approach is introduced which will extent traditional association study settings gaining better understanding of complex diseases like hypertension and might give better chances to evaluate association studies for their functional relevance. PMID- 16472130 TI - Nutraceuticals--an emerging era in the treatment and prevention of cardiovascular diseases. AB - Foods and nutrients play a vital role in normal functioning of the body. They are helpful in maintaining the health of the individual and in reducing the risk of various diseases. Worldwide acceptance of this fact formed a recognition link between "nutrition" and "health" and the concept of "nutraceuticals" was evolved. Nutraceuticals are medicinal foods that play a role in maintaining well being, enhancing health, modulating immunity and thereby preventing as well as treating specific diseases. Thus the field of nutraceuticals can be envisioned as one of the missing blocks in the health benefit of an individual. More than any other disease, the etiology of cardiovascular disease reveals many risk factors that are amenable to nutraceutical intervention. The scientific literature shows that several ingredients marketed for use in dietary supplements address each of these. The ability of nutraceuticals to positively influence cardiovascular risk factors should be recognized as an enormous opportunity in the treatment of a highly prevalent disease. Nutraceuticals hold promise in clinical therapy as they have the potential to significantly reduce the risk of side effects associated with chemotherapy along with reducing the global health care cost. In this review, an attempt has been made to summarize some of the recent research findings on garlic, omega-3-fatty acids, soy products, dietary fibres, vitamins, antioxidants, plant sterols, flavonoids, prebiotics and probiotics that have beneficial effects on the heart, in order to update the practising clinician on the benefit of using nutraceuticals for the management of cardiovascular diseases. PMID- 16472131 TI - ADAMTS proteinases: potential therapeutic targets? AB - ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) proteinases have been implicated in a number of connective tissue pathologies including Ehlers-Danlos syndrome type VII C, Weill-Marchesani syndrome, encephalomyelitis, and arthritis. These proteinases therefore represent potential therapeutic targets for the treatment of such conditions. The synthesis and activity of ADAMTS proteinases is regulated at multiple levels: transcription, RNA splicing, translation, proteolytic processing, cofactor stimulation and inhibition, each of which represents a possible point of therapeutic intervention. Recent research suggests that, in addition to the direct inhibition of ADAMTS proteinases with low molecular weight non-peptidic inhibitors, targeting the transcription and protein processing of these enzymes could be effective therapeutic approaches. PMID- 16472133 TI - Fluorescence fluctuation spectroscopy in reduced detection volumes. AB - Fluorescence fluctuation spectroscopy is a versatile technique applied to in vitro and in vivo investigations of biochemical processes such as interactions, mobilities or densities with high specifity and sensitivity. The prerequisite of this dynamical fluorescence technique is to have, at a time, only few fluorescent molecules in the detection volume in order to generate significant fluorescence fluctuations. For usual confocal fluorescence microscopy this amounts to a useful concentration in the nanomolar range. The concentration of many biomolecules in living cell or on cell membranes is, however, often quite high, usually in the micro- to the millimolar range. To allow fluctuation spectroscopy and track intracellular interaction or localization of single fluorescently labeled biomolecules in such crowded environments, development of detection volumes with nanoscale resolution is necessary. As diffraction prevents this in the case of light microscopy, new (non-invasive) optical concepts have been developed. In this mini-review article we present recent advancements, implemented to decrease the detection volume below that of normal fluorescence microscopy. Especially, their combination with fluorescence fluctuation spectroscopy is emphasized. PMID- 16472132 TI - Micropropagation: a tool for the production of high quality plant-based medicines. AB - Medicinal plants are the most important source of life saving drugs for the majority of the world's population. The biotechnological tools are important to select, multiply and conserve the critical genotypes of medicinal plants. Plant tissue culture techniques offer an integrated approach for the production of standardized quality phytopharmaceutical through mass-production of consistent plant material for physiological characterization and analysis of active ingredients. Micropropagation protocols for cloning of some medicinal plants such as Catharanthus roseus (Apocynaceae), Chlorophytum borivilianum (Liliaceae), Datura metel (Solanaceae), and Bacopa monnieri (Scrophulariaceae) have been developed. Regeneration occurred via organogenesis and embryogenesis in response to auxins and cytokinins. The integrated approaches of our culture systems will provide the basis for the future development of novel, safe, effective, and high quality products for consumers. PMID- 16472134 TI - Membrane channels as therapeutic targets. PMID- 16472135 TI - Exploiting high-throughput ion channel screening technologies in integrated drug discovery. AB - Ion channels are increasingly being implicated in disease. Although existing drugs that modulate channel function currently represent a key class of pharmaceutical agents, future ion channel drugs could help to treat an even wider variety of diseases. Despite their disease relevance, ion channels remain largely under exploited as drug targets, chiefly resulting from the absence of screening technologies that provide the throughput and quality of data required to support medicinal chemistry. Although some technical challenges still lie ahead, this historic bottleneck in drug discovery is now being bypassed by newer technologies that can be fully integrated into the early stages of drug discovery and will allow the discovery of novel therapeutic agents. Sequencing the human genome has greatly added to the number of potential drug targets but selecting suitable ion channels for drug discovery research should be based on the potential therapeutic relevance of the channel and not just the availability of suitable screens. Currently, ion channel drug discovery is focused on the need to identify compounds that can provide tractable starting points for medicinal chemistry. Advances in laboratory automation have brought significant opportunities to increase screening throughput for ion channel assays but careful assay configuration to model drug-target interactions in a physiological manner remains an essential consideration. Ion channel screening platforms are described in this review to provide some insight into the variety of technologies available for screening, together with some of their inherent advantages and limitations. PMID- 16472136 TI - Brain neuronal nicotinic receptors as new targets for drug discovery. AB - Neuronal nicotinic receptors (nAChRs) are a heterogeneous family of ion channels differently expressed in the nervous system where, by responding to the endogenous neurotransmitter acetylcholine, they contribute to a wide range of brain activities and influence a number of physiological functions. Over recent years, the application of newly developed molecular and cellular biological techniques has made it possible to correlate the subunit composition of nAChRs with specific nicotine-elicited behaviours, and refine some of the in vivo physiological functions of nAChR subtypes. The major new findings are the widespread expression of nAChRs, outside the nervous system, their specific and complex organisation, and their relevance to normal brain function. Moreover, the combination of clinical and basic research has better defined the involvement of nAChRs in a growing number of nervous pathologies other than degenerative diseases. However, there are still only a limited number of nicotinic-specific drugs and, although some nicotinic agonists have an interesting pharmacology, their clinical use is limited by undesirable side effects. Some selective nicotinic ligands have recently been developed and used to explore the complexity of nAChR subtype structure and function in the expectation that they will become rational therapeutic alternatives in a number of neurodegenerative, neuropsychiatric and neurological disorders. In this review, we will discuss the molecular basis of brain nAChR structural and functional diversity mainly in pharmacological and biochemical terms, and summarise current knowledge concerning the newly discovered drugs used to classify the numerous receptor subtypes and treat the brain diseases in which nAChRs are involved. PMID- 16472137 TI - Na+ channel pharmacology and molecular mechanisms of gating. AB - Electrogenesis of efficiently propagated action potentials requires synchronized opening of transmembrane Na+ channels possessing a sodium selectivity-filter, a high-throughput ion-conductance pathway, and voltage-dependent gating functions. These properties of the Na+ channel have long been the target of molecular analysis. Several toxins and drugs, known to selectively bind to Na+ channels, have been used as pharmacological tools to investigate Na+ channel properties either electrophysiologically or chemically. Recent analyses of the protein crystal structure of bacterial voltage-dependent K+ channels have provided important clues to the identity of mobile structures involved in channel gating. The new information may be applicable to Na+ channels, and may well require a total revision of our understanding of gating mechanisms of sodium channels. Several experiments challenge the emerging view that channel gating by S6 transmembrane segments is triggered by signals from voltage sensors floating in membrane lipid. Herein, we review the various toxin and drug molecules that affect the gating behavior of Na+ channels in this new structural framework, by characterizing the binding sites of these toxins, and assessing the pharmacological effects resulting from changes in the structure of the toxin or sodium channel. PMID- 16472138 TI - L-type calcium channels. AB - The Ca2+ channel blockers represent a successful group of therapeutic agents directed against cardiovascular targets, including hypertension and angina. These drugs, including the first-generation verapamil, nifedipine and diltiazem are directed against a subclass of voltage-gated Ca2+ channel - the L-type channel. Other subclasses of Ca2+ channel exist and are targets for new indications. The mechanisms of actions of the L-type blockers are discussed and the origins of their cardiovascular selectivity discussed. Although new drugs of this class directed against hypertension could be developed, there are both clinical and economic reasons that argue against such development. However, there are other possible targets to investigate where antagonists and activators of the L-type channel may be useful: such targets include fertility, neuronal growth, bone formation and epilepsy. Limitations to these approaches are discussed. PMID- 16472139 TI - Modulation of potassium channels as a therapeutic approach. AB - Regulation of potassium (K+) channels evokes hyperpolarization or repolarization of the cell membrane to prevent or reverse cell excitability and is fundamental in the control of cellular activity throughout the range of tissue types within the human body. Genome projects predict that in excess of 80 K+ channel-related genes exist, resulting in a high degree of K+ channel diversity. In addition, dysfunction of K+ channels, as a result of mutations of the genes for the channel proteins or alterations in channel regulation, has been associated with the pathophysiology of diseases. These observations support K+ channels as therapeutic targets to regulate cellular homeostasis in pathophysiological conditions. Molecular cloning and expression of K+ channels offer important information in the identification of selective compounds to provide unique tissue management. Specific modulators have been identified for a limited number of K+ channel subtypes. Unfortunately the conversion of data obtained in the laboratory to success in the clinical setting has been limited. Tissue delivery of genes, in combination with drugs, may be an avenue enabling specific modulation of ion channel function and improved drug selectivity. Using specific examples (HERG, IKs, KCNQs, KCa, Kv1.3), issues regarding distribution, function and diversity related to advances made in the identification of modulators having therapeutic potential are discussed. The scope of this field is just emerging and the number of likely therapeutic indications for K+ channel modulators will increase as insight into the dynamics of expression of these channels in various diseases grows and the issue of the required selectivity is resolved. PMID- 16472140 TI - On the discovery and development of CFTR chloride channel activators. AB - Chloride channels play important roles in vital cellular signalling processes contributing to homeostasis in both excitable and non-excitable cells. Since 1987, more than ten ion channel genes have been identified as causing human hereditary diseases among them the genes for the voltage-dependent chloride channel ClC-1 (myotonia) and the cystic fibrosis transmembrane conductance regulator (CFTR) protein (cystic fibrosis). The CFTR gene was cloned in 1989 and its protein product identified as an ATP-gated and phosphorylation-regulated chloride channel during the following two years. Since then, searching for potent and specific small molecules able to modulate normal and mutated CFTR has become a crucial endpoint in the field for both our understanding of the physiological role that CFTR plays in epithelial cells and more importantly for the development of therapeutic agents to cure cystic fibrosis (CF). It is predicted that a pharmacological approach would help not only to restore the defective transport activity of mutant CFTR but also to correct the regulatory function of CFTR. This review describes the evolution of CFTR pharmacology and how during the last five years, high throughput screening assays have been developed to identify novel molecules, some of them probably constituting a reservoir of future therapeutic agents for CF. PMID- 16472141 TI - Membrane ion channels and diabetes. AB - Type-2, or non-insulin-dependent diabetes mellitus is a serious disease that is now widespread throughout Western society. Glucose intolerance, or failure of glucose to stimulate insulin secretion, is a primary factor in the manifestation of this disease and is likely to be due to the failure of glucose metabolism to stimulate pancreatic beta-cell electrical activity, calcium influx, and insulin secretion. In this review we describe how ion channels regulate the electrical behaviour of the beta-cell and how the membrane potential depolarises in response to a rise in glucose metabolism. Central to these electrical events is the inhibition of ATP-sensitive potassium channel by ATP, and we summarise recent advances in our understanding of the properties of this ion channel in coupling beta-cell metabolism to electrical activity. We discuss the mechanism, specificity, and clinical implications of the pharmacological inhibition of KATP channels by sulphonyureas and other antidiabetic drugs. The roles of other ion channels in regulating electrical activity are considered, and also their potential use as targets for drug action in treating beta-cell disorders. PMID- 16472142 TI - Recent progress in pharmacological and non-pharmacological treatment options of major depression. AB - In spite of recent progress in the pharmacotherapy of depression major issues are still unresolved. These include the non-response rate of approximately 30% to conventional antidepressant pharmacotherapy, side effects of available antidepressants and the latency of several weeks until clinical improvement. The only non-pharmacological biological treatment options available so far which exert more rapid antidepressant efficacy are electroconvulsive therapy and, as an augmentation strategy, sleep deprivation. Current pharmacological treatments aim to enhance serotonergic and/or noradrenergic neurotransmission. In spite of emerging knowledge, the crucial mechanisms underlying both non-pharmacological treatments, which are responsible for antidepressant efficacy, are not yet clear so far. In the meantime several new pharmacological principles are under investigation with regard to their putative antidepressant potency. These include 5-HT1A receptor agonists, tachykinin receptor antagonists and various interventions within the hypothalamic-pituitary-adrenal system. While there is evidence for antidepressant properties of these new treatments in animal studies, in case series, in open studies and to some degree also in placebo controlled studies, no definite proof for the antidepressant efficacy of these new pharmacological strategies according to the requirements for evaluation of antidepressant drugs has been furnished so far. In contrast, for the established non-pharmacological treatment strategies including bright light therapy the clinical efficacy has been proven at least in subgroups of depression, but more knowledge of the main mechanisms underlying their antidepressant efficacy is still necessary. In addition new non-pharmacological treatments like repetitive transcranial magnetic stimulation, magnetic seizure therapy and Vagus nerve stimulation are currently under development. Nevertheless, a follow-up of both the new pharmacological strategies and non-pharmacological treatment options is of major importance to provide even better strategies for the clinical management of depression, which also is of great socio-economic impact. PMID- 16472143 TI - Does Angiotensin converting enzyme inhibitor protect the heart in cardiac surgery? From laboratory to operating room: clinical application of experimental study. AB - Animal studies have shown angiotensin converting enzyme (ACE) inhibitors to be effective agents for myocardial protection. They protect against lethal arrhythmias, preserve ventricular function, improve coronary reserve (especially after ischemia/reperfusion), and reverse myocardial hypertrophy. Human studies, on the other hand, have shown inconsistent results. The beneficial effects of ACE inhibitors demonstrated in animal studies provide major advantages for cardiac surgery. First, most cardiac surgery is performed under ischemic arrest induced by a cardioplegic solution, and the protective effects of ACE inhibition against reperfusion injury can reduce peri-operative mortality and morbidity. Second, most patients who undergo such surgery have myocardial hypertrophy due to hypertension, pressure or volume overload mediated by valve disease, or myocardial infarction. Ventricular hypertrophy is a strong risk factor for sudden death, probably from arrhythmia. Regression of the hypertrophy may prevent post operative sudden death, thereby allowing for long-term benefits of surgery. In this paper, I review ACE inhibitor studies in animals and humans and the protective mechanisms involved. I also discuss why human studies show inconsistent results in spite of the fact that ACE inhibition is consistently protective in animal studies. Finally, I explore the potential clinical applications of ACE inhibitors in cardiac surgery. PMID- 16472145 TI - Potential disease targets for drugs that disrupt protein-- protein interactions of Grb2 and Crk family adaptors. AB - This review summarises some of the knowledge we have about Crk and Grb2 family adaptor protein signalling in health and disease and outlines the current status and the challenges still remaining in the development of efficient and selective inhibitors of their protein - protein interactions. It also highlights briefly some recent successes and problems of inhibitors for proteins that functionally interact with Crk and Grb2 family adaptors, as well as opportunities, which may arise from combination therapies. Grb2 and Crk family adaptors regulate signalling pathways linked to human diseases. They are mainly composed of Src homology 2 (SH2) and Src homology 3 (SH3) domains, which serve as docking sites for signalling proteins, including various receptors, cytoplasmic kinases and GTPase regulators. Considerable insight into the biological functions and mechanisms of action of small SH2/SH3 domain adaptors has been gained in the last years from experimental approaches as diverse as targeted gene disruption and structural studies at the atomic level. This has already indicated several strategies to utilise SH2 and SH3 domain interaction inhibitors in human disease therapy. Additional molecular targets for Crk and Grb2 domain interaction blockers are expected to surface as further protein-protein interactions are discovered. Examples include newly found DOCK family proteins (DOCK3, DOCK4, and DOCK5) which are known or suspected effectors of Crk proteins and the interaction of Grb2 with the cell cycle regulator p27Kip1. PMID- 16472146 TI - Gene therapy for primary immunodeficiency diseases: recent progress and misgivings. AB - The progress of clinical gene therapy trials during the last two decades has been remarkable, and its application has also expanded into various fields of human diseases. Among them, hereditary diseases such as the primary immunodeficiency diseases (PID) were considered suitable candidates for gene therapy because the therapeutic strategy was very simple, therefore, effective gene therapy may be obtained without significant difficulty compared to other more complex diseases such as cancer. Indeed, the first clinical gene therapy trial was safely performed and was in part, effective for adenosine deaminase (ADA) deficiency patients, a type of severe combined immunodeficiency diseases (SCID). However, because of certain unforeseen obstacles, it took approximately 10 years until the first curative effects were obtained for gene therapy in patients with X-linked SCID (X-SCID). Here, I review and discuss the background and historical events leading up to PID gene therapy, the safety issues, which unexpectedly arose after the successful report, and finally I will attempt to predict the future trends in this form of gene therapy. PMID- 16472147 TI - Application of hematopoietic cells to therapeutic angiogenesis. AB - Despite considerable progress in the field of cardiovascular medicine and surgery, ischemic heart disease is still the leading cause of death in advanced countries. In this context, it is no wonder why therapeutic angiogenesis, a way to ameliorate ischemic tissue from suffering dysfunction by increasing new blood vessels, gains so much attention from both clinicians and patients. In this review, we will briefly go through a decade of history in therapeutic angiogenesis including unraveling of its mechanisms, results obtained from clinical trials, and lessons learned from earlier investigations. We will then focus on an emerging, yet rapidly evolving field of hematopoietic cell therapy. Recent excellent studies seem to have brought us to the place where we might save so many patients from burden of ischemia, we should be aware that there are some controversies, and sometimes misunderstandings, regarding how or why this treatment does actually work, and what better way should we explore in order to get the best of its efficacy. With these caveats in mind, we will investigate the works elucidating the mechanisms and clinical efficacies of hematopoietic cell therapy. PMID- 16472148 TI - Cilostazol: therapeutic potential against focal cerebral ischemic damage. AB - Cilostazol was developed as a selective inhibitor of cyclic nucleotide phosphodiesterase 3 (PDE3). The anti-platelet and vasodilator properties of cilostazol have been extensively characterized and considered to contribute to the variety of clinical effects such as intermittent claudication and recurrent stroke. In this review, the novel action mechanism (s) of cilostazol are overviewed with the focus on the action of cilostazol in in vitro and in vivo studies as a maxi-K channel opener targeting anti-apoptotic signaling pathways. Under treatment with cilostazol (10 mg/kg intravenously or 30 mg/kg orally), a significant reduction in cerebral infarct area was evident in rats subjected to ischemia/reperfusion. Increase in cyclic AMP and decrease in TNF-alpha levels were identified in the ipsilateral cortex under treatment with cilostazol accompanied by decreased Bax formation and cytochrome c release with increased Bcl-2 production in the penumbral area as well as in the in vitro human umbilical endothelial cells. Cilostazol suppressed TNF-alpha-induced decrease in viability of SK-N-SH (human neuroblastoma) cells and HCN-1A (human cortical neuron) cells in association with decrease in PTEN phosphorylation and increase in Akt/CREB phosphorylation with suppression of DNA fragmentation, all of which were antagonized by iberiotoxin, a maxi-K(+) channel blocker. Further, cilostazol prevented TNF-alpha-induced PTEN phosphorylation and apoptotic cell death via increased CK2 phosphorylation in the SK-N-SH cells. Cilostazol increased K(+) current in SK-N-SH cells by opening the maxi-K channels. Thus, it was suggested that the action of cilostazol to promote cell survival was ascribed to the maxi-K channel opening-coupled upregulation of CK2 phosphorylation and downregulation of PTEN phosphorylation with resultant increased phosphorylation of Akt and CREB. These in vitro data were confirmed in the in vivo results of rats subjected to focal transient ischemic damage. PMID- 16472149 TI - Sphingosine 1-phosphate in vascular biology: possible therapeutic strategies to control vascular diseases. AB - Blood platelets are very unique in that they store sphingosine 1-phosphate (Sph-1 P) abundantly (possibly due to the existence of highly active sphingosine kinase and a lack of Sph-1-P lyase) and release this bioactive lipid extracellularly upon stimulation. Vascular endothelial cells (ECs) and smooth muscle cells (SMCs) respond dramatically to this platelet-derived bioactive lipid mainly through a family of G protein-coupled Sph-1-P receptors named S1P1, 2, 3, 4, and 5, originally referred to as EDG-1, 5, 3, 6, and 8, respectively. In fact, the importance of Sph-1-P in platelet-vascular cell interactions has been revealed in a number of recent reports. Through interaction with ECs, Sph-1-P can mediate physiological wound healing processes such as vascular repair, although this important bioactive lipid can become atherogenic and thrombogenic, and cause or aggravate cardiovascular diseases especially under certain pathological conditions. On the other hand, Sph-1-P induces vasoconstriction through interaction with SMCs. It is likely that regulation of Sph-1-P biological activities is important for the therapeutical purpose to control vascular disorders. Particularly, the development of specific S1P receptor agonists or antagonists seems a reasonable strategy to selectively regulate the bioactivity of Sph-1-P, considering that a great diversity of Sph-1-P actions has been reported and that this diversity depends mainly on the S1P receptor subtype involved. In this review, I will summarize recent findings on possible roles of Sph-1-P in vascular biology and its therapeutical implications. PMID- 16472150 TI - Pulmonary surfactant proteins A and D: innate immune functions and biomarkers for lung diseases. AB - Pulmonary surfactant, a complex of lipids and proteins, functions to keep alveoli from collapsing at expiration. Surfactant proteins A (SP-A) and D (SP-D) belong to the collectin family and play pivotal roles in the innate immunity of the lung. Pulmonary collectins directly bind with broad specificities to a variety of microorganism and possess anti-microbial effects. These proteins also exhibit both inflammatory and anti-inflammatory functions, which occur through interactions with pattern recognition receptors including Toll-like receptor and CD14, signal inhibitory regulatory protein alpha and a receptor complex of calreticulin and CD91. The collectins enhance phagocytosis of microbes by macrophages through opsonic and/or non-opsonic activities. The proteins stimulate cell surface expression of phagocytic receptors including scavenger receptor A and mannose receptor. Since the expression of SP-A and SP-D is abundant and restricted within the lung, the proteins are now clinically used as biomarkers for lung diseases. The levels of SP-A and SP-D in bronchoalveolar lavage fluids, amniotic fluids, tracheal aspirates and pleural effusions reflect alterations in alveolar compartments and epithelium, and lung maturity. The determination of SP A and SP-D in sera is a non-invasive and useful tool for understanding some pathological changes of the lung in the diseases, including pulmonary fibrosis, collagen vascular diseases complicated with interstitial lung disease, pulmonary alveolar proteinosis, acute respiratory distress syndrome and radiation pneumonitis. PMID- 16472151 TI - Antioxidant vitamins and risk of lung cancer. AB - Tobacco use is the leading risk factor for lung cancer, yet in addition to smoking habit, diet may also play a role in the disease's appearance. While there are reports to indicate that antioxidant vitamins and carotenoids may decrease the risk of lung cancer, results to date have been somewhat ambiguous. This review aimed to describe the results yielded by different studies, which have addressed antioxidant vitamin intake and lung cancer, and to indicate the mechanisms whereby these nutrients might be exercising their activity. Antioxidant vitamins were observed to have no clear protective effect, though there was some evidence pointing to a protective role for vitamins C and E. Vitamin A, in contrast, evinced no clear effect. Insofar as provitamin A carotenoids were concerned, lutein/zeaxanthin, lycopene and alpha-carotene displayed a certain protective trend, yet beta-carotene exhibited no protective effect whatsoever; and indeed, there was speculation as to whether it might even be pernicious in smokers. Beta-criptoxanthin, on the other hand, showed a more consistent protective effect. The study highlighted the need to conduct further research on smokers and non-smokers alike, and in particular, to investigate the effect, if any, on lung cancer of carotenoids or vitamins when ingested in differing dosages. PMID- 16472152 TI - Calpain inhibition: a therapeutic strategy targeting multiple disease states. AB - The calpains represent a well-conserved family of calcium-dependent cysteine proteases. They consist of several ubiquitous and tissue specific isoforms and exhibit broad substrate specificity influencing many aspects of cell physiology including migration, proliferation and apoptosis. Calpain activity in vivo is tightly regulated by its natural endogenous inhibitor calpastatin. Calpastatin specifically inhibits calpain and not other cysteine proteases by interaction with several sites on the calpain molecule. Inappropriate regulation of the calpain-calpastatin proteolytic system is associated with several important human pathological disorders including muscular dystrophy, cancer, Alzheimer's disease, neurological injury, ischaemia/reperfusion injury, atherosclerosis, diabetes and cataract formation. Recent advances in elucidating the tertiary structures of calpain 2 and its regulatory domain calpain 4, together with identification of new modes of regulating calpain activity provide new opportunities for the design of novel calpain inhibitors. Several classes of inhibitors, including peptidyl epoxide, aldehyde, and ketoamide inhibitors, targeting the active site have proven effective against the calpains and are in the process of evaluation in animal models of human disease. However, a major limitation to the clinical use of such inhibitors is their lack of specificity among cysteine proteases and other proteolytic enzymes. The development of a new class of calpain inhibitors that interact with domains outside of the catalytic site of calpain may provide greater specificity and therapeutic potential. PMID- 16472153 TI - Advances in chondroitin sulfate analysis: application in physiological and pathological States of connective tissue and during pharmacological treatment of osteoarthritis. AB - Recent glycobiology studies have suggested fundamental biological functions for chondroitin sulfate (CS) and dermatan sulfate (DS), which are widely distributed as glycosaminoglycans (GAGs) sidechains of proteoglycans (PGs) in the extracellular matrix and at cellular level. Several biological functions are closely associated with the structure and in particular with the sulfation patterns of these polysaccharides. CS is also used as a structure-modifying osteoarthritis (OA) drug that reverses, retards, or stabilizes the pathology of OA, thereby providing symptomatic relief in the long-term treatment. Advances in analytical separational techniques, including agarose-gel electrophoresis, high performance liquid chromatography (HPLC), capillary electrophoresis (HPCE), fluorophore-assisted carbohydrate electrophoresis (FACE) and electrospray ionization mass (ESI-MS) enable us to examine alterations of CS/DS with respect to their quantities and fine structural features in various pathological conditions, thus becoming applicable for diagnosis. Furthermore, sensitive analytical procedures enable us to follow the pharmacological application of CS in the treatment of OA and to monitor the progression of the disorder. In this review, the chromatographic and electromigration procedures developed to analyse and characterise CS/DS are presented. Moreover, a critical evaluation of the biological relevance of the results obtained by the developed methodology is discussed. PMID- 16472155 TI - gamma-secretase as a therapeutic target for treatment of Alzheimer's disease. AB - Alzheimer's disease (AD) is the most common cause of dementia with aging, that is pathologically characterized by senile plaques that contain amyloid-beta peptides (Abeta) and neurofibrillary tangles comprised of phosphorylated tau. Genetic and biological studies provide evidence that the production and deposition of Abeta contribute to the etiology of AD. gamma-Secretase is the pivotal enzyme in generating the C terminus of Abeta, that determines its aggregability and propensity for deposition. Drugs that regulate the production of Abeta by inhibiting gamma-secretase activity could provide an effective therapeutics for AD, although recent studies suggest that gamma-secretase plays important roles in novel signaling pathways that play essential roles in embryonic development. This review focuses on recent progresses in the gamma-secretase biology that shed substantial light on the proteolytic mechanism, regulation and composition of this unusual enzyme. Moreover, we review the recent development of inhibitors and provide a direction for the effective treatment of AD through inhibition of gamma secretase activity. PMID- 16472156 TI - Reduction of Abeta levels in the Sprague Dawley rat after oral administration of the functional gamma-secretase inhibitor, DAPT: a novel non-transgenic model for Abeta production inhibitors. AB - Considerable effort has been made to develop drugs that delay or prevent neurodegeneration. These include inhibitors of Abeta-generating proteases for the treatment of Alzheimer's disease. Testing the amyloid hypothesis in vivo requires molecules that are capable of entering the CNS and that produce a substantial reduction in brain Abeta levels. Plaque-developing APP transgenic mice are currently widely used as an in vivo model of choice as these animals produce readily measurable amounts of human Abeta. They are very useful in the testing of a variety of amyloid-lowering approaches but their use for compound screening is often limited by their cost. Transgenic animals also require extensive, time consuming breeding programs and can show high inter-animal differences in the expression level of the transgene. Hence, we considered it important to develop and characterize a new and simple non-transgenic animal model for testing Abeta modulation. For this purpose, Wild-type adult Sprague Dawley rats were treated with DAPT, a functional gamma-secretase inhibitor, and the Abeta40 and Abeta42 levels in brain-tissue and body fluids were assessed. We showed that DAPT, given orally, significantly lowered Abeta40 and Abeta42 peptide levels in brain extract, CSF, and the plasma dose- and time-dependently. We can conclude that our data establish the usefulness of the wild-type rat model for testing small molecule inhibitors of Abeta production. PMID- 16472157 TI - Targeting the alpha 7 nicotinic acetylcholine receptor to reduce amyloid accumulation in Alzheimer's disease pyramidal neurons. AB - Although there is still no known effective preventative treatment or cure for Alzheimer's disease (AD), the development of new drugs that target pathological features that appear early in the course of this disease and alleviate some of the early cognitive and memory symptoms is a laudable goal that may be one step closer. To date, the acetylcholinesterase inhibitors have been the most widely used AD drugs and have been somewhat successful in slowing loss of cognition. In the last few years, a number of studies have demonstrated that amyloid beta (1 42) (Abeta42), the predominant Abeta peptide species in amyloid plaques, first accumulates in vulnerable neurons prior to plaque formation. Recently, we have shown that many (if not most) amyloid plaques in the entorhinal cortex of AD brains are actually the lysis remnants of degenerated, Abeta42-overburdened neurons. Furthermore, the most vulnerable neurons appear to be those that abundantly express the alpha7 nicotinic acetylcholine receptor (alpha7nAChR), and internalization of Abeta42 appears to be facilitated by the high-affinity binding of Abeta42 to the alpha7nAChR on neuronal cell surfaces, followed by endocytosis of the resulting complex and its accumulation within the lysosomal compartment. This mechanism provides a reasonable explanation for the selective vulnerability of cholinergic and cholinoceptive neurons in AD brains and for the fact that Abeta42 is the dominant Abeta peptide species in both intraneuronal accumulations and amyloid plaques. In view of the pathophysiological consequences of Abeta42 binding to alpha7nAChR on neuronal surfaces that stem from excessive intraneuronal Abeta42 accumulation, the alpha7nAChR could be an important therapeutic target for treatment of AD. In addition, it further emphasizes the potential merits of new and effective therapeutic strategies pointed towards the goal of lowering of Abeta42 levels in the blood and cerebrospinal fluid as well as blocking Abeta42 in the blood from penetrating the blood-brain barrier and entering into the brain parenchyma. PMID- 16472158 TI - Antigonadotropins: a novel strategy to halt Alzheimer's disease progression. AB - A significant amount of research has been focused on the relationship between hormones and Alzheimer's disease. However, the majority of this work has been on estrogen and more recently testosterone. A serendipitous patient encounter led one of us (RLB) to question whether other hormones of the hypothalamic-pituitary gonadal axis could be playing a role in the pathogenesis of Alzheimer's disease. The age-related decline in reproductive function results in a dramatic decrease in serum estrogen and testosterone concentrations and an equally dramatic compensatory increase in serum luteinizing hormone concentrations. Indeed, there is growing evidence that the gonadotropin, luteinizing hormone, which regulates serum estrogen and testosterone concentrations, could be an important causative factor in the development of Alzheimer's disease. This review provides information supporting the "gonadotropin hypothesis," puts forth a novel mechanism of how changes in serum luteinizing hormone concentrations could contribute to the pathogenesis of Alzheimer's disease, and discusses potential therapeutic anti-gonadotropin compounds. PMID- 16472159 TI - The cell cycle and hormonal fluxes in Alzheimer disease: a novel therapeutic target. AB - Several hypotheses have been proposed attempting to explain the pathogenesis of Alzheimer disease (AD) including theories involving amyloid deposition, tau phosphorylation, oxidative stress, metal ion dysregulation and inflammation. Strong evidence suggests that each one contributes to disease pathogenesis, though none of these mechanisms result in all the downstream changes that occur during the course of AD. For this reason, we and others have begun the search for a causative factor that predates known features found in AD, and that might be a fundamental initiator of the pathophysiological cascade. In this regard, we propose that the dysregulation of the cell cycle that occurs in neurons susceptible to degeneration in the hippocampus during AD is a potential causative factor that would initiate all known pathological events. Neuronal changes supporting alterations in cell cycle control in the etiology of AD include the ectopic expression of markers of the cell cycle, organelle kinesis and cytoskeletal alterations including tau phosphorylation. Given the early and presumably devastating consequences of cell cycle re-entry, we have made a concerted effort to elucidate the initiating factor that drives aberrant mitotic re-entry in AD. As a result of the gender bias present in AD, we suspect that postmenopausal and andropausal hormones may be involved and, with this in mind, in this review we specifically focus on the gonadotropins. Therapeutic interventions targeted at gonadotropins, if they are indeed the driving mitogenic force, could both prevent disease in those patients currently asymptomatic or halt, and even reverse, disease in those currently afflicted. PMID- 16472160 TI - Antioxidants as a potential therapy against age-related neurodegenerative diseases: amyloid Beta toxicity and Alzheimer's disease. AB - Alzheimer's disease (AD) is a progressive age-related neurodegenerative disorder with distinct neuropathological features. Extracellular plaques, consisting of aggregated amyloid peptides of 39-43 amino acids are one of the most prominent pathological hallmarks of this disease. Although the exact neurochemical effector mechanism of Abeta aggregation is not yet elucidated, age-associated disturbances of metal ion metabolism have been proposed to promote the formation of aggregates from soluble Abeta. Oxidative stress is postulated to be a downstream effect of Abeta-metal ion interactions. Therefore, the modulation of brain metal metabolism and attenuation of oxidative stress by antioxidant molecules are proposed as a potential therapeutic intervention in AD. Here, we summarize the recent literature focused on APP/Abeta-metal ion interactions and the use of antioxidant metal chelators as potential therapy against AD. PMID- 16472161 TI - Neuroprotective therapies for Alzheimer's disease. AB - One of the major age-related damaging agents are reactive oxygen species (ROS). The brain is more vulnerable to oxidative stress than other organs as concomitant low activity and capacity of antioxidative protection systems allow for increased exposure of target molecules to ROS. Since neurons are postmitotic cells, they have to live with cellular damage accumulated over many decades. Increased levels of ROS (also termed "oxidative stress"), produced by normal mitochondrial activity, inflammation and excess glutamate levels, are proposed to accelerate neurodegenerative processes characteristic of Alzheimer's disease. This review presents evidence of the importance of oxidative stress in the pathogenesis of these diseases and explains the nature of different types of ROS mediating neuronal damage. Furthermore, the potential beneficial effects of neuroprotective treatments, including antioxidants and anti - glutamatergic drugs are discussed. PMID- 16472162 TI - Brain inflammation, cholesterol, and glutamate as interconnected participants in the pathology of Alzheimer's disease. AB - Alzheimer's disease (AD) represents one of the most common ailments afflicting the rapidly growing elderly segment of today's population. Despite the vast amount of effort expended in developing a cure, currently approved drugs address only cognitive symptoms that, although important for improving a patient's daily living standard, do not provide a significant delay or halt to disease progression. Early reports that individuals taking anti-inflammatory medications reduce their risk of developing AD has led to the "inflammation hypothesis" of AD and the subsequent testing of these drugs in the clinic. Tests of a select few of these medications in AD clinical trials have, however, yielded disappointing results. Reports of statin-based medications reducing the risk of AD have also led to the testing of this class of drugs in the clinic. Recently, the approval of the NMDA receptor antagonist memantine (Namenda) has provided clinical support for glutamatergic processes in the disease and generated a renewed interest in the role of excitatory amino acids in the etiology of AD. In this review, we take a closer look at these three compelling areas for addressing AD therapeutics: inflammation, cholesterol, and glutamate. We present arguments that these components are interconnected and mutually regulate processes involved in AD progression. Special focus is given to inflammation as a central feature of AD that may be acting in synergy with cholesterol and glutamate to mediate the observed pathophysiology. PMID- 16472163 TI - Mitochondria: a target for neuroprotective interventions in cerebral ischemia reperfusion. AB - Evidence obtained over the past two decades shows that reactive oxygen species (ROS) are involved in brain lesions, including those due to cerebral ischemia reperfusion. The mitochondria are the primary intracellular source of ROS, as they generate huge numbers of oxidative-reduction reactions and use massive amounts of oxygen. When anoxia is followed promptly by reperfusion, the resulting increase in oxygen supply leads to overproduction of ROS. In ischemic tissues, numerous studies have established a direct role for ROS in oxidative damage to lipids, proteins, and nucleic acids. Thus, mitochondria are both the initiator and the first target of oxidative stress. Mitochondrial damage can lead to cell death, given the role for mitochondria in energy metabolism and calcium homeostasis, as well as the ability of mitochondria to release pro-apoptotic factors such as cytochrome C and apoptosis-inducing factor (AIF). This review discusses possible mitochondrion-targeted strategies for preventing ROS-induced injury during reperfusion. The sequence of events that follow oxidative damage provides the outline for the review: thus, we will discuss protection of oxidative phosphorylation, mitochondrial membrane integrity and fluidity, and antioxidant or mild-uncoupling strategies for diminishing ROS production. Among mechanisms of action, we will describe the modulation of mitochondrial permeability transition pore (MPTP) opening, which may not only operate as a physiological Ca(2+) release mechanism, but also contribute to mitochondrial deenergization, release of pro-apoptotic proteins, and protection by ischemic preconditioning (IPC). Finally, we will review genetic strategies for controlling apoptotic protein expression, stimulating mitochondrial oxidative defences, and increasing mitochondrial proliferation. PMID- 16472164 TI - Fatty acid amide hydrolase: a potential target for next generation therapeutics. AB - Endocannabinoids are amides, esters and ethers of long chain polyunsaturated fatty acids, which act as new lipid mediators. Anandamide (N arachidonoylethanolamine; AEA) and 2-arachidonoylglycerol are the main endogenous agonists of cannabinoid receptors, able to mimic several pharmacological effects of Delta(9)-tetrahydrocannabinol, the active principle of Cannabis sativa preparations like hashish and marijuana. The activity of AEA at its receptors is limited by cellular uptake through a specific membrane transporter, followed by intracellular degradation by a fatty acid amide hydrolase (FAAH). Growing evidence demonstrates that FAAH is the critical regulator of the endogenous levels of AEA, suggesting that it may serve as an attractive therapeutic target for the treatment of human disorders. In particular, FAAH inhibitors may be next generation therapeutic drugs of potential value for the treatment of pathologies in the central nervous system and in the periphery. Here, the potential applications of these inhibitors for human disease will be reviewed, with an emphasis on the properties of hydro(pero)xy-anandamides. In fact, these oxygenated derivatives of AEA are the most powerful inhibitors of FAAH of natural origin as yet discovered. In addition, new insights into the promoter region of FAAH gene will be presented, and the therapeutic potential of mimetics of transcription factors of this gene in the management of human infertility will be discussed. PMID- 16472166 TI - On the existence of a global molecular network enmeshing the whole central nervous system: physiological and pathological implications. AB - Proteins are endowed with the "Lego property", i.e., the capability of steric fitting with other proteins to form high molecular weight complexes with emergent functions. These interactions may occur both as horizontal molecular networks at the plasma membrane level and as vertical molecular networks, i.e., towards the extra- and/or intracellular side of the cell. The present paper broadens this view by proposing the existence of three dimensional molecular networks, mainly made by proteins and carbohydrates, which might interact with each other at boundaries of compartments such as plasma membranes to form a "global molecular network" (GMN) that pervades the intra- as well as the extra-cellular environment of the entire central nervous system. The GMN is a potentially plastic structure regulated through several means. For example, its extra-cellular part is under the remodeling action of the matrix metalloproteinases. The proposal of a GMN has physiological and pathological implications. In primis, classical synaptic transmission, gap junctions and volume transmission signals by modulating GMN could importantly contribute to the "binding phenomenon", i.e. the phase synchronization of firing rates in far-located neuronal cortical groups. Secondly, alterations in protein conformation could alter the GMN organization and hence the neuronal network morphology and function. This could lead to the formation of abnormal protein aggregates such as amyloid plaques and neurofibrillary tangles, which, in turn, might affect the GMN function and/or the reciprocal interactions between its parts especially at the boundaries between compartments. PMID- 16472167 TI - The allosteric properties of hemoglobin: insights from natural and site directed mutants. AB - After over a century of extensive research, hemoglobin has become the prototype of allosteric and cooperative proteins. Its molecular structure, known in great detail, has allowed the design of hundreds of site directed mutations, aimed at interfering with its function, and thus at testing our hypotheses on the molecular mechanisms of allostery. The wealth of information thus obtained is difficult to read except for specialists, not only because it makes use of many different technical approaches, but also because of its intrinsically patchy nature. Moreover, several researchers have tried to assign specific roles to segments of the polypeptide chains, rather than to single residues, and have tested their hypotheses by multiple point mutations or by complete replacement with the homologous segment from a different hemoglobin to produce chimeric macromolecules. This approach is in great need of a revision since putative functionally relevant segments partially overlap. This review briefly describes the structure and function of hemoglobin, and analyzes the effect of point mutations, multiple mutations and segment replacement, with special attention to possible biotechnological applications, ranging from pharmacology (Hb solutions as resuscitating fluids and sources of the protein found in hemoglobinopathies for biochemical studies) to bioreactors. Occasional reference is made to site directed mutants of myoglobin, whenever this helps clarifying perplexing results obtained on hemoglobin. PMID- 16472168 TI - Expression of highly toxic genes in E. coli: special strategies and genetic tools. AB - Escherichia coli (E. coli) remains the most efficient widely-used host for recombinant protein production. Well-known genetics, high transformation efficiency, cultivation simplicity, rapidity and inexpensiveness are the main factors that contribute to the selection of this host. With the advent of the post-genomic era has come the need to express in this bacterium a growing number of genes originating from different organisms. Unfortunately, many of these genes severely interfere with the survival of E. coli cells. They lead to bacteria death or cause significant defects in bacteria growth that dramatically decrease expression capabilities. In this paper, we review special strategies and genetics tools successfully used to express, in E. coli, highly toxic genes. Suppression of basal expression from leaky inducible promoters, suppression of read-through transcription from cryptic promoters, tight control of plasmids copy numbers and proteins production as inactive (but reversible) forms are among the solutions presented and discussed. Special expression vectors and modified E. coli strains are listed and their effectiveness illustrated with key examples, some of which are related to our study of the highly toxic phage T4 restriction endoribonuclease RegB. We mainly selected those strategies and tools that permit E. coli normal growth until the very moment of highly toxic gene induction. Expression then occurs efficiently before cells die. Because they do not target a particular toxic effect, these strategies and tools can be used to express a wide variety of highly toxic genes. PMID- 16472169 TI - Recombinant allergens for diagnosis and immunotherapy of allergic disorders, with emphasis on cockroach allergy. AB - The prevalence of allergic disorders has increased over the past few decades and the quality of life has been significantly influenced at least for the allergic subjects. Allergen avoidance is thought to be the best way of preventing clinical manifestation of the disease, however, it is not possible for some allergens, and other pharmacological and/or immunological treatment has to be made. Repetitive injection of sensitized allergens to the patients (immunotherapy) is the only known curative approach to the disease even though the exact mechanism is not clear to date. Crude extract of allergens has lots of shortcomings which might arouse unexpected results. Genetic engineering and recombinant allergens are thought to be one of the alternative ways to overcome these limitations. Genetic engineering could facilitate the investigation of immune responses of the subjects especially on B cell and T cell epitopes, and produce the therapeutic allergens which might minimize the possible side effects. Furthermore, conjugation of immuno-modulatory molecules such as CpG-ODN, cytokines, or toxins which could act specifically to the given allergens, and maleylation of the allergens could maximize the prophylactic or therapeutic effect. Immunotherapies for the pollen allergy and insect sting allergy have been thought to be successful. House dust mite allergy and cockroach allergy have been reported less beneficial by immunotherapeutic approaches. Cockroaches are one of the most important causes of asthma, and severe complications are often reported in the children in city dwellers with low-incomes. The studies of the biological functions of cockroach allergens and the use of recombinant allergens should allow understanding of mechanisms of cockroach-elicited allergic disorders and development of allergen-specific and sensitive diagnostics and tailored therapeutic approaches in the future. PMID- 16472170 TI - Survey of functional activities of alpha-fetoprotein derived growth inhibitory peptides: review and prospects. AB - Alpha-fetoprotein (AFP), known largely as a growth-promoting agent, possesses a growth-inhibitory motif recently identified as an occult epitopic segment in the third domain. The present study reviews the multiple biological activities of this AFP-derived peptide segment termed the Growth Inhibitory Peptide (GIP), which is a 34-amino acid fragment taken directly from the full-length 590 amino acid molecule. The GIP segment has been chemically synthesized, purified, characterized, and subjected to a variety of bioassays. The GIP has a proven record of growth suppression in both fetal and tumor cells, but not in normal adult cells. Even though the mechanism of action has not been completely elucidated, GIP participates in various biological activities such as endocytosis, angiogenesis, and cytoskeleton-induced/cell shape changes. In this review, a survey of the functional roles of the GIP is presented which encompasses multiple organizational levels of GIP involvement, including the 1) organism, 2) organ, 3) tissue, 4) cell, 5) plasma membrane, 6) cytoplasm, and 7) the nucleus. At the cell membrane interface, the actions of GIP are discussed concerning cell aggregation, agglutination, adhesion, and migration in light of GIP serving as a possible decoy ligand and/or soluble receptor. Regarding cytosolic activities, GIP has been reported to inhibit various cytoplasmic enzyme activities, modulate apoptotic events, and regulate cytoplasmic signal transduction (MAP kinase) cascades. Concerning the nuclear compartment, GIP is capable of complexing with the estrogen receptor and binding estradiol, but does not affect estradiol-induced estrogen receptor transcription. In overview, efforts were made to review the multiple biological activities reported for GIP in order to prioritize likely physiological activities and present an updated consensus of functional roles for this AFP-derived peptide. PMID- 16472171 TI - The effects of olprinone, a phosphodiesterase 3 inhibitor, on systemic and cerebral circulation. AB - Olprinone, a phosphodiesterase (PDE) 3 inhibitor, is used to treat heart failure due to its positive inotropic and vasodilative effects. Selective inhibition of the PDE 3 isozyme increases intracellular adenosine 3;5;-cyclic monophosphate and enhances Ca(2+) influx into cardiac muscle cells. The most significant advantage of PDE 3 inhibitors is their ability not only to enhance myocardial contraction, but to reduce, through vasodilatory action, the stress to which the heart is subjected. In peripheral vessels, the decrease of cytosolic free Ca(2+) induces the vasorelaxation of vascular smooth muscle cells. In this way, olprinone reduces mean aortic and pulmonary artery pressures. Additionally, olprinone exerts differential vasodilatory effects on peripheral vessels in each organ, based on the differences in the distribution of PDE 3 among the organs. With respect to the cerebral circulation, olprinone augments blood flow in the cerebral cortex through direct vasodilatory effects on small cerebral arteries or arterioles. Olprinone increases hepatosplanchnic blood flow and improves oxygen supply. While long-term therapy with PDE 3 inhibitors in patients with chronic heart failure may accelerate the progress of the underlying disease and provoke serious ventricular arrhythmia, olprinone shows good potential for short-term treatment in patients who have experienced severe heart failure or patients who have undergone cardiac surgery. PMID- 16472172 TI - The influence of sex hormones on pulmonary vascular reactivity: possible vasodilator therapies for the treatment of pulmonary hypertension. AB - Pulmonary hypertension is a rare disease of the pulmonary vasculature defined as a mean pulmonary artery pressure >25 mmHg at rest or 30 mmHg with exercise. Recent therapies such as epoprostenol, bosentan and sildenafil are directed at the arterial vascular bed, causing vasodilatation and reducing pulmonary vascular resistance. However idiopathic pulmonary artery hypertension (IPAH) occurs predominantly in women, with three times the incidence compared to men and this suggests that sex hormones may be involved in the pathogenesis. 17beta oestradiol is a pulmonary vasodilator, proposed to act via an endothelium dependant pathway, involving nitric oxide (NO) and has also been shown to alter responses to hypoxia. Progesterone is also a pulmonary vasodilator but differs from 17beta-oestradiol in having endothelial-dependant and independent processes implicated. Interestingly testosterone has been shown to be a vasodilator in both the coronary and pulmonary circulation with a mechanism of action involving calcium channel blockade of the vascular smooth muscle and without endothelial involvement. In clinical trials testosterone confers symptomatic benefits in patients with coronary heart disease and heart failure, acting as a vasodilator. These observations lend support to the notion that testosterone could be a potential treatment for patients with PAH as vasodilator therapy remains the mainstay of treatment. Other potential beneficial effects of testosterone in the pulmonary circulation include immuno-modulation, altering expression of cytokines and an anti-thrombotic action. In this review the influence of sex hormones on the pulmonary vasculature will be discussed, with specific focus on pulmonary hypertension and the potential treatment of this condition. PMID- 16472173 TI - Hydrogen sulfide as a new endogenous gaseous transmitter in the cardiovascular system. AB - Hydrogen sulfide (H(2)S) is a well-known toxic gas with the smell of rotten eggs. Recent studies have shown that H(2)S is generated in vivo in human and animal organisms and that it participates in many pathophysiological processes. H(2)S is produced endogenously in mammalian tissues from L-cysteine metabolism mainly by 3 enzymes: cystathionine beta-synthetase (CBS), cystathionine gamma-lyase (CSE) and 3-mercaptosulfurtransferase (MST). H(2)S may not only function as a neuromodulator in the central nervous system but it also relaxes gastrointestinal smooth muscles. More importantly, present evidence shows that H(2)S exerts regulatory effects on the pathogenesis of various cardiovascular diseases such as hypertension, pulmonary hypertension, shock and myocardial injury. The genomic basis of cystathioninuria in humans is 2 nonsense and 2 sense mutations in CSE. This review reveals that H(2)S is a new endogenous gaseous transmitter in the cardiovascular system. PMID- 16472174 TI - Chronic inhibition of na(+)/h(+)-exchanger in the heart. AB - The incidence and prevalence of heart failure (HF) has increased over the last decades. The main reasons for this increase are the ageing population and an increase in survival rate after myocardial infarction and other cardiovascular diseases. Although, pharmacotherapy has significantly improved survival, the prognosis of HF is still rather poor. Total mortality is high and approximately half of the deaths are sudden and unexpected. Angiotensin-converting-enzyme (ACE) inhibitors generally given with diuretic and digoxin are the standard treatment for patients with HF. Despite the established benefits of ACE-inhibitors there is a need for new pharmacological tools for the treatment of HF. Recent experimental evidence has shown that activity of the Na(+)/H(+)-exchanger in the heart (NHE-1) is increased in HF. Because NHE-1 exchanges intracellular H(+) for extracellular Na(+) in a one by one stoichiometry, the intracellular ionic changes resulting from increased activity, will be a increased pH(i) and intracellular sodium ([Na(+)](i)). Activation of NHE-1 results only in a small increase in pH(i), under physiological conditions where bicarbonate-dependent mechanisms are active. However, a considerable increase in [Na(+)](i) was always present. The elevation of [Na(+)](i) might be responsible for the increase of intracellular calcium ([Ca(2+)](i)) levels mediated by the Na(+)/Ca(2+)-exchanger (NCX). Increases in [Na(+)](i), pH(i) and [Ca(2+)](i), features of cardiac myocytes isolated from failing hearts, are recognized as a cell growth signal And thus may play a role in the hypertrophic response, cellular remodeling and finally the development of HF. Acute application of cariporide, an inhibitor of NHE-1, on failing myocytes not only normalized [Na(+)](i) but also cytoplasmic and sarcoplasmic reticulum calcium handling and the propensity to develop delayed after depolarizations (DAD's). In several animal models of HF it has been shown the chronic inhibition of NHE-1 attenuates the development of hypertrophy and whole heart remodeling. Recently, in a volume and pressure overload model of HF in rabbits it has been demonstrated that chronic treatment also prevents the development of HF and cellular ionic and electrophysiological remodeling. Therefore, chronic treatment with an inhibitor of NHE-1 might prove beneficial in patients at risk of developing HF, especially when given at an early stage. PMID- 16472175 TI - Thrombolytic therapy in acute ischemic stroke - basic concepts. AB - Thrombolytic therapy with alteplase in acute ischemic stroke is currently established within 3 h from symptom onset in carefully selected patients. Expansion of the time window is being assessed in trials with alteplase and desmoteplase. Also, tenecteplase, reteplase and staphylokinase are being evaluated in stroke patients. A better understanding of the coagulation and fibrinolytic systems may provide insight into drug development, interactions, complications, and may ultimately improve patient triage and treatment regimens. Future adjuvant and additional therapies could prove useful for patients who do not benefit from standard thrombolytic treatment or may augment the overall gain. PMID- 16472176 TI - Towards newer molecular targets for chronic diabetic complications. AB - Prior to the discovery of insulin, the major cause of death in the diabetic population was ketoacidosis. Although insulin and improved glycemic control have improved the longevity of diabetic patients, they still suffer from significant morbidity and mortality due to chronic secondary complications. Long standing diabetes leads to structural and functional alterations in both the micro- and macrovasculature. These complications, involving the retina, kidney, and peripheral nerves, as well as cardiovascular system, severely compromise the quality and expectancy of life. Large scale clinical trials have identified hyperglycemia as the key determinant for the development of such complications. Therapeutic modalities have been developed to target glucose-induced alterations, such as protein kinase C activation, augmented polyol pathway activity, non enzymatic glycation and oxidative stress to ameliorate chronic complications. However, clinical trials targeting these biochemical alterations have failed to show significant beneficial effects. The plethora of biochemical anomalies that govern the development of chronic diabetic complications may therefore be subject to cross-interaction and complex interplays. Studies in both animal and human diabetes have, however, showed alteration of several vasoactive effector molecules such as endothelins. These molecules may be instrumental in mediating diabetes-induced structural and functional deficits at both the early and late stages of the disease. This review will discuss the current mechanistic understanding of chronic diabetic complications and will explore the potential novel therapeutic interventions. PMID- 16472177 TI - Therapeutic potential of endothelial progenitor cells for cardiovascular diseases. AB - In the past decade, researchers have defined committed stem or progenitor cells from various tissues, including bone marrow, peripheral blood, brain, liver and reproductive organs, in both adult animals and humans. Recently, endothelial progenitor cells (EPCs) were isolated from peripheral blood mononuclear cells and were shown to be incorporated into foci of neovascularization. This finding that circulating EPCs may home into sites of neovascularization and differentiate into mature endothelial cells in situ is consistent with the concept of 'vasculogenesis' and suggests that vasculogenesis and angiogenesis might constitute complementary mechanisms for postnatal neovascularization. Furthermore, experimental and clinical studies on ischemic cardiovascular diseases suggest a therapeutic potential for EPC transplantation. In this review, we summarize the biological features of EPCs and discuss their therapeutic potential for the treatment of cardiovascular diseases. PMID- 16472178 TI - Angiotensin II regulates vascular and endothelial dysfunction: recent topics of Angiotensin II type-1 receptor signaling in the vasculature. AB - Accumulating evidence strongly implicates angiotensin II (AngII) intracellular signaling in mediating cardiovascular diseases such as hypertension, atherosclerosis and restenosis after vascular injury. In vascular smooth muscle cells (VSMCs), through its G-protein-coupled AngII Type 1 receptor (AT(1)), AngII activates various intracellular protein kinases, such as receptor or non-receptor tyrosine kinases, which includes epidermal growth factor receptor (EGFR), platelet-derived growth factor receptor (PDGFR), c-Src, PYK2, FAK, JAK2. In addition, AngII activates serine/threonine kinases such as mitogen-activated protein kinase (MAPK) family, p70 S6 kinase, Akt/protein kinase B and various protein kinase C isoforms. In VSMCs, AngII also induces the generation of intracellular reactive oxygen species (ROS), which play critical roles in activation and modulation of above signal transduction. Less is known about endothelial cell (EC) AngII signaling than VSMCs, however, recent studies suggest that endothelial AngII signaling negatively regulates the nitric oxide (NO) signaling pathway and thereby induces endothelial dysfunction. Moreover, in both VSMCs and ECs, AngII signaling cross-talk with insulin signaling might be involved in insulin resistance, an important risk factor in the development of cardiovascular diseases. In fact, clinical and pharmacological studies showed that AngII infusion induces insulin resistance and AngII converting enzyme inhibitors and AT(1) receptor blockers improve insulin sensitivity. In this review, we focus on the recent findings that suggest the existence of novel signaling mechanisms whereby AngII mediates processes, such as activation of receptor or non-receptor tyrosine kinases and ROS, as well as cross-talk between insulin and NO signal transduction in VSMCs and ECs. PMID- 16472179 TI - The effects of obesity-related peptides on the vasculature. AB - Obesity and its related metabolic diseases, including type 2 diabetes, are associated with alterations in the circulating levels of various peptides. These include the adipocytokines (peptides released by adipocytes which circulate, such as leptin, adiponectin and resistin), and other peptides whose levels are altered in association with obesity (such as ghrelin, neuropeptide Y, interleukin-1 beta and tumour necrosis factor-alpha). While the primary action of these peptides is linked with the regulation and maintenance of energy balance and metabolism, many of them have also been shown to possess vasoactive, inflammatory and other properties that influence vascular biology, vascular physiology and atherogenesis. As such, they may form an important mechanistic link between obesity and cardiovascular disease. In this review, we will outline the vasoactive properties of adipocytokines and other obesity-related peptides. In particular, as pharmacotherapies suggested to achieve weight loss will alter the pathways associated with these peptides, such treatments might have either beneficial or deleterious effects on the incidence and progression of cardiovascular disease. PMID- 16472180 TI - Probing for antiparasitic drugs. PMID- 16472181 TI - Functional cloning as a means to identify Leishmania genes involved in drug resistance. AB - Resistance to anti-leishmanial drugs is a mounting problem in high-endemicity regions of South Asia and, potentially, in the context of HIV-Leishmania coinfections in Southern Europe. The molecular basis for clinical drug resistance is still largely unknown. It is important, however, to identify all relevant drug resistance markers for further drug development and for epidemiological surveys. An elegant and powerful method to identify such drug resistance markers without bias is functional cloning, using cosmid-based genomic DNA libraries. This review discusses the merits and caveats of this approach. PMID- 16472182 TI - The use of anticancer drugs in antiparasitic chemotherapy. AB - Many similarities exist between cancer cells and parasites. A potentially lucrative starting point for the discovery of novel drugs to combat parasites is to examine available compounds developed against cancer for antiparasitic properties. Here, we review the use of current and promising anticancer agents for treating major human parasitic diseases. PMID- 16472183 TI - Development of miltefosine for the leishmaniases. AB - The leishmaniases consist of visceral and cutaneous syndromes present in > 30 endemic regions of the world. Miltefosine (hexadecylephosphocholine) is the first oral agent that is effective and tolerated for both visceral and cutaneous disease in several endemic regions, and represents a major advance in the treatment of these diseases. PMID- 16472184 TI - New formulations and derivatives of amphotericin B for treatment of leishmaniasis. AB - The clinical treatment of leishmaniasis is based on a limited number of drugs, which are associated with adverse effects and have already induced resistance. Amphotericin B (AmB), a polyene antibiotic produced by Streptomyces sp, is the only anti-leishmanial drug which has not induced clinical resistance since its discovery in 1956. The limiting factor in the use of AmB is its toxic effects, mainly nephrotoxicity. The maximal dose of AmB for human use is 1.5 mg/kg which sometimes is not sufficient for cure. The mode of action of AmB is associated with its toxicity: it selectively binds to parasite membrane ergosterol but also, to a lesser extent, to human cholesterol. Apart from this mechanism, AmB has immunomodulatory effects, some of them are deleterious. Reduction of the toxic effects by using lipid formulations allows the infusion of higher doses of AmB. Unfortunately, these formulations are relatively expensive and therefore out of reach for patients in need, in the endemic areas. All the existing formulations are given parenterally, which has obvious disadvantages; most important is the need for hospitalization or multiple visits in the clinic. The current efforts to improve AmB are directed at the production of AmB aggregates in liquid solutions, encapsulation with lipid components, and solubilization by binding to soluble polymers. The expected improved treatment resulting from use of the new formulations is based on better pharmacokinetics, reduced toxicity originating from slow release, targeting to the infected organ and an altered pattern of immune responses (related to AmB). Of particular importance are the attempts to produce derivatives for oral treatment, which will decrease costs of hospitalization and improve applicability for children and the elderly population. PMID- 16472185 TI - Interference with redox-active enzymes as a basis for the design of antimalarial drugs. AB - Antimalarial drugs are urgently and continuously required. Parasite enzymes involved in antioxidant defence represent interesting target molecules for rational drug development. Here we summarize the currently available data on structural, biochemical, and functional properties of these proteins in an attempt to evaluate and compare their potential as drug targets. PMID- 16472186 TI - Drug Targets for Plasmodium falciparum: a post-genomic review/survey. AB - Over 300 million cases of malaria each year cause significant morbidity and mortality. Growing drug-resistance among the Plasmodia that cause malaria motivates the development of additional anti-malarial drugs. This review summarizes the current state of knowledge about potential drug targets for malaria. The recently sequenced malaria genome data clarifies parasite metabolic pathways, and more metabolic targets have been identified. PMID- 16472187 TI - Antibiotics which target the Wolbachia endosymbionts of filarial parasites: a new strategy for control of filariasis and amelioration of pathology. AB - Wolbachia endosymbionts of filariae are targets for the development of new antifilarial chemotherapy. Doxycycline to deplete Wolbachia from the worm has demonstrated the feasibility of this strategy and has provided a new chemotherapeutic tool. Recent research shows that depleting Wolbachia will also lessen pathology, and lessen adverse reactions to traditional antifilarial drugs. PMID- 16472188 TI - Applications of electron spin resonance and spin trapping in tropical parasitic diseases. AB - Free radicals may be reaction intermediates in biological systems in more situations than are presently recognized. However, progress in detecting such species by Electron Spin Resonance (ESR) has been relatively slow. ESR is a very sensitive technique for free radical detection and characterization. It can be used to investigate very low concentrations of radicals provided that they are stable enough for their presence to be detected. For unstable radicals special techniques have to be employed. One of these methods is called Spin Trapping. Parasitic diseases in tropical and subtropical areas constitute a major health and economic problem. The range of antiparasitic drugs varies widely in structural complexity and action at the subcellular and molecular levels. However, a number of these drugs are thought to exert their action by generating free radicals. Most of the free radical producing drugs used against parasites are: quinones, naphtoquinones, quinone-imines, aminoquinolines, N-oxides and nitroheterocyclic compounds. This review summarizes some of the more relevant achievements of ESR and Spin Trapping applications in parasitic diseases studies. The use of ESR spectroscopy to obtain relevant information about free radical characterization and the analysis of the mechanisms of action of drugs involved in several parasitic diseases is also presented. PMID- 16472189 TI - Thyrotropin-releasing hormone analogs. AB - Thyrotropin releasing hormone (TRH: pyroglutamic acid-histidine-prolineamide) regulates the activity of cells in the anterior pituitary and within the central and peripheral nervous systems. TRH, which has been the subject of much research over the past three decades, exerts its effects by acting through class A G protein coupled receptors. The recent discovery of a second receptor subtype has generated an interest in the discovery of receptor subtype-selective TRH analogs. In this review, we describe advances in the development of TRH analogs and in the understanding of their mechanism of interaction with TRH receptors. We also describe the recent breakthrough in the identification of analogs that bind selectively at TRH-R2. PMID- 16472190 TI - Multimodality imaging of tumor integrin alphavbeta3 expression. AB - Most solid tumors are angiogenesis dependent. Anti-angiogenic pharmaceuticals that inhibit the growth of new blood vessels offer considerable promise as anti cancer agents. With increasing numbers of anti-angiogenic drugs in clinical trials, there is an urgent need for detailed characterization of the heterogeneity of tumor vasculature and dissection of the complex network of mechanisms that control tumor angiogenesis. Non-invasive molecular imaging will play a key role in individualized anti-angiogenic therapy based upon molecular features of the new blood vessel growth. Integrin alpha(v)beta(3), which binds several ligands via an RGD tripeptide sequence, is uniquely expressed in tumor vasculature and aggressive tumor cells, making it a potential target for anti angiogenic interventions. This review highlights some recent advances in multimodality imaging of tumor integrin expression with emphasis on positron emission tomography (PET). PMID- 16472191 TI - Antioxidants as novel agents for asthma. AB - Oxidative stress plays an important role in the pathogenesis of asthma. Recently several investigators have studied the effects of a variety of antioxidants on asthma. Antioxidants, including L-2-oxothiazolidine-4-carboxylic acid, reduce airway inflammation and hyper-responsiveness of asthma and may be novel therapeutic agents for asthma. PMID- 16472192 TI - Proteomics in medicinal chemistry. AB - Proteomics is becoming an important research area for studying protein expression patterns induced by different external stimuli. An important aspect of proteomics is to identify and quantify proteins. Many new technologies and techniques have been developed in this field and have been applied to various aspects of drug discovery. PMID- 16472195 TI - Neurogenesis as a therapeutic strategy for cognitive aging and Alzheimer's disease. PMID- 16472196 TI - Small molecule approaches for promoting neurogenesis. AB - The discovery of small molecules capable of promoting neurogenesis will contribute to the elucidation of the physiological roles of neurogenesis and to novel therapeutic approaches. Small molecule development can be targeted to the promotion of precursor proliferation, survival, migration or maturation and might be applied to augmenting physiological neurogenesis already present in the dentate gyrus or subventricular zone/olfactory bulb or to normally non-neurogenic regions relevant to neuropathological states. Current small molecule discovery can be assessed from the perspective of the following categories: compounds modulating physiological signaling pathways regulating neurogenesis including the sonic hedgehog, bone morphogenic protein Wnt/,-catenin, Notch and chemokine systems; growth factor mimetics; protein tyrosine phosphatase inhibitors; existing drugs including antidepressants, lithium, valproate, sildenafil and statins; hormones, steroids and peptides; and neurotransmitter receptor agonists and antagonists. Unbiased, high throughput screening will likely lead to the discovery of additional active compounds and the recognition of novel mechanisms regulating neurogenesis. A major therapeutic challenge will consist of the identification of molecular targets and mechanisms relatively specific for precursor cells of interest. PMID- 16472197 TI - Preclinical analyses of the therapeutic potential of allopregnanolone to promote neurogenesis in vitro and in vivo in transgenic mouse model of Alzheimer's disease. AB - Herein, we present data to support a preclinical proof of concept for the therapeutic potential of allopregnanolone to promote neurogenesis. Our recent work has demonstrated that the neuroactive progesterone metabolite, allopregnanolone (3alpha-hydroxy-5alpha-pregnan-20-one), (APalpha) induced, in a dose dependent manner, a significant increase in proliferation of neuroprogenitor cells (NPCs) derived from the rat hippocampus and human neural stem cells (hNSM) derived from the cerebral cortex [1]. Proliferative efficacy was determined by incorporation of BrdU and (3)H-thymidine, FACS analysis of MuLV-GFP-labeled mitotic NPCs and quantification of total cell number. Allopregnanolone-induced proliferation was isomer and steroid specific, in that the stereoisomer 3beta hydroxy-5beta-pregnan-20-one and related steroids did not increase (3)H-thymidine uptake. Immunofluorescent analyses for the NPC markers, nestin and Tuj1, indicated that newly formed cells were of neuronal lineage. Furthermore, microarray analysis of cell cycle genes and real time RT-PCR and western blot validation revealed that allopregnanolone increased the expression of genes which promote mitosis and inhibited the expression of genes that repress cell proliferation. Allopregnanolone-induced proliferation was antagonized by the voltage gated L-type calcium channel blocker nifedipine consistent with the finding that allopregnanolone induces a rapid increase in intracellular calcium in hippocampal neurons via a GABA type A receptor activated L-type calcium channel. Preliminary in vivo data indicate that APalpha for 24 hrs significantly increased neurogenesis in dentate gyrus, as determined by unbiased stereological analysis of BrdU positive cells, of 3-month-old male triple transgenic Alzheimer's disease mice. The in vitro and in vivo neurogenic properties of APalpha coupled with a low molecular weight, easy penetration of the blood brain barrier and lack of toxicity, are key elements required for developing APalpha as a neurogenic / regenerative therapeutic for restoration of neurons in victims of Alzheimer's disease. PMID- 16472198 TI - Dissecting the diverse actions of pro- and mature neurotrophins. AB - The neurotrophins mediate diverse actions in the developing peripheral and central nervous systems. They are initially synthesized as precursor forms, or proneurotrophins, that are cleaved to release C-terminal mature forms that bind to Trk receptor tyrosine kinases to enhance synaptic plasticity and neuronal survival. Recent studies suggest that proneurotrophins are not inactive precursors, but signaling proteins that can activate the p75 receptor to mediate diverse responses. Proneurotrophins can activate a heteromeric receptor complex of p75 and sortilin to initiate cell death, or bind to p75 in hippocampal neurons to enhance long term depression. Thus, neurotrophin actions are regulated by the form of the neurotrophin (pro- or mature) secreted by cells, by extracellular proteolytic cleavage of proneurotrophins to generate mature forms, and by the expression of neurotrophin receptors Trk, or p75 and sortilin, that are selectively activated by mature or proneurotrophins, respectively. Here, recent studies are reviewed that reveal that pro- and mature neurotrophins have distinct and sometimes opposing actions in regulating cell death and survival in development and in pathophysiologic states, in regulating neurotrophin secretion, and in modulating synaptic plasticity. PMID- 16472199 TI - Neurodegeneration and neurogenesis: focus on Alzheimer's disease. AB - Neurogenesis, or the production of new neurons from neuronal precursor cells, is a normal phenomenon in the adult brain, and is accentuated by brain injury. Forms of injury associated with increased neurogenesis include both acute (e.g., stroke) and chronic neurodegenerations. Studies on human postmortem material and transgenic mice overexpressing amyloid precursor protein mutations found in familial Alzheimer's disease (AD) suggest that AD is associated with enhanced neurogenesis. However, the mechanism responsible for this effect is unknown, as is what influence it may have on the clinical course of murine or human AD. If AD leads to the production of fully functional, mature neurons that can restore brain function, strategies aimed at further increasing endogenous neurogenesis may have therapeutic value. PMID- 16472201 TI - Effects of paliroden (SR57667B) and xaliproden on adult brain neurogenesis. PMID- 16472200 TI - VEGF, a mediator of the effect of experience on hippocampal neurogenesis. AB - Rodents housed in an enriched environment, exercise by running or perform learning and memory tasks show an increase in hippocampal neurogenesis. We show that both environmental enrichment, as well as performance in the Morris water maze, a hippocampal-dependent learning task, leads to an increase in local VEGF expression in rats. We genetically recreated this situation by somatic cell gene transfer using recombinant adeno-associated virus (AAV) vectors. Genetically increasing hippocampal VEGF in adult rats resulted in a approximately 2 fold increase in neurogenesis associated with improved cognition. In contrast, gene transfer of placental growth factor (PGF) which signals through Flt1, but not KDR receptors had negative effects on neurogenesis and inhibited learning, although it similarly increased endothelial cell proliferation. Expression of a dominant negative, mKDR, inhibited basal neurogenesis and impaired learning. Co-expression of mKDR antagonized VEGF-enhanced neurogenesis and learning without inhibiting endothelial cell proliferation. Furthermore, inhibition of VEGF expression by RNA interference completely blocked the environmental induction of neurogenesis. These data support a model whereby VEGF acting via KDR is a mediator of the effect of the environment on neurogenesis and cognition [1]. PMID- 16472202 TI - Implications for CNS repair of redox modulation of cell survival, division and differentiation. AB - Studies on oligodendrocytes, the myelin-forming cells of the central nervous system, and on the progenitor cells from which they are derived, have provided several novel insights into the role of intracellular redox state in cell function. A central unifying theme of this research is that redox state modulation lies at the heart of understanding cell-intrinsic aspects of precursor cell function, responsiveness of precursor cells to cell-extrinsic signals and even the means by which cell-extrinsic signaling molecules alter cellular function. This review discusses our studies on the role in redox state as a critical modulator of cellular function, and considers the implications of these findings for optimizing tissue repair. PMID- 16472203 TI - Environment, physical activity, and neurogenesis: implications for prevention and treatment of Alzhemier's disease. AB - Age is the biggest risk factor for the development of neurodegenerative diseases. Consequently, as the population ages it becomes more critical to find ways to avoid the debilitating cost of neurodegenerative diseases such as Alzheimer's. Some of the non-invasive strategies that can potentially slow down the mental decline associated with aging are exercise and use of multi-sensory environmental stimulation. The beneficial effects of both exercise and multi-sensory environmental stimulation have been well-documented, thus it is possible that these strategies can either provide neuroprotection or increase resistance to the development of age-related cognitive problems. PMID- 16472204 TI - Discovery of neurogenic, Alzheimer's disease therapeutics. AB - Many researchers have questioned whether new potential therapies aimed at reversing Alzheimer's disease (AD) are indeed scientifically feasible. A number of approved therapies already exist for Alzheimer's disease, yet these drugs only slow the disease progression for a period of time and treat the symptoms of this devastating disease. New therapies intended to reverse the disease would necessarily need to replace dead, dying and dysfunctional neurons in affected regions of the brain. This complex drug discovery problem is further complicated by the knowledge that AD is mainly an aging disorder and that aging, though not considered a disease, causes biological changes that may also need to be overcome [1]. The requirement for new, functional neurons under neurodegenerative diseases, as seen in AD and stroke suggests that an inhibitor of neuronal death, like Memantine, is insufficient to reverse the cognitive and physical loss. New neurons, or neurogenesis, may be required for real improvement or reversal of the cognitive deficit. Adult neurogenesis, first described by Altman in the early 1960s [2, 3], has more recently been observed as a response to injury or disease. Of interest was the finding that new neurons appear to migrate to disease/injury affected areas in the brain not normally neurogenic in the adult. This pathological-stimulation of neurogenesis does not appear sufficient to stave off the disease and subsequent behavioral decline. Therefore, the desire to amplify and improve upon the neurogenesis-response to neurodegenerative disease appears warranted, if not yet feasible. The key to doing so lies in identifying what signals are required to promote neurogenesis and neuron survival, either in injury and disease or under environmental stimuli. This could provide clues for how to pharmacologically induce neurogenesis under neurodegenerative conditions. Currently, progress in identifying therapeutics that appear to promote ameliorative neurogenesis for AD is lagging behind the pharmacological induction of neurogenesis as a therapy for depression. PMID- 16472205 TI - sAPPalpha enhances the transdifferentiation of adult bone marrow progenitor cells to neuronal phenotypes. AB - The remediation of neurodegeneration and cognitive decline in Alzheimer's Disease (AD) remains a challenge to basic scientists and clinicians. It has been suggested that adult bone marrow stem cells can transdifferentiate into different neuronal phenotypes. Here we demonstrate that the alpha-secretase-cleaved fragment of the amyloid precursor protein (sAPPalpha), a potent neurotrophic factor, potentiates the nerve growth factor (NGF)/retinoic acid (RA) induced transdifferentiation of bone marrow-derived adult progenitor cells (MAPCs) into neural progenitor cells and, more specifically, enhances their terminal differentiation into a cholinergic-like neuronal phenotype. The addition of sAPPalpha to NGF/RA-stimulated MAPCs resulted in their conversion to neuronal like cells as evidenced by the extension of neurites and the appearance of immature synaptic complexes. MAPCs differentiated in the presence of sAPPalpha and NGF/RA exhibited a 40% to as much as 75% increase in neuronal proteins including NeuN, beta-tubulin III, NFM, and synaptophysin, compared to MAPCs differentiated by NGF/RA alone. This process was accompanied by an increase in the levels of choline acetyltransferase, a marker of cholinergic neurons, compared to those of GABAergic and dopaminergic neuronal subtypes. MAPCs immunopositive for sAPPalpha were identified within the septohippocampal system of transgenic PS/APP mice injected intravenously with sAPPalpha-transfected MAPCs and found in close proximity to the cerebral vasculature. Given that in AD cholinergic neurons are severely vulnerable to neurodegeneration and that the levels of sAPPalpha are significantly reduced, these findings suggest the combined use of sAPPalpha and MAPCs offers a new and potentially powerful therapeutic strategy for AD treatment. PMID- 16472206 TI - Has the amyloid cascade hypothesis for Alzheimer's disease been proved? AB - After much initial debate for and against the role of amyloid in Alzheimer's disease (AD), mutations on the amyloid precursor protein (APP) and processing pathways that increase levels of the amyloid b peptide of 42 residues (Abeta42) have established that faulty function or processing of these proteins are responsible for AD pathogenesis. Given the neurotoxicity of aggregates of Ab42, the central role of this peptide in AD pathogenesis is self evident. In this article, I summarize the major pieces of evidence adduced to support the amyloid cascade hypothesis and point out their limitations. PMID- 16472207 TI - Amyloid beta: the alternate hypothesis. AB - Alzheimer disease (AD) is a devastating condition and patients, caregivers, clinicians, and scientists are eager to decipher the underlying disease mechanism and, thereafter, target this therapeutically. Most investigators studying the underlying cause of AD have focused on amyloid-beta (Abeta) such that the Amyloid Cascade Hypothesis is the predominant mechanism thought to be responsible for the disease. However, a number of caveats have led us to seriously question the validity of this hypothesis. First, in addition to increases in Abeta, genetic mutations in AD lead to increased vulnerability to oxidative/apoptotic insults indicating that the mutated protein disturbs redox balance. Whether mutations result in Abeta deposition that then causes oxidative stress or whether mutations cause oxidative stress that results in Abeta deposition is unclear. Indeed, while in vitro experiments show that Abeta can directly cause oxidative stress to cells in culture, it is apparent from other studies that the reverse is also true, namely that oxidative stress leads to increases in Abeta. Notably, in vivo studies in both sporadic and genetic forms of the disease show that oxidative stress temporally precedes increases in Abeta and that increases in Abeta are associated with a decrease in oxidative stress. Based on these findings, we herein propose an Alternate Amyloid Hypothesis in which pathogenic factors for disease lead to increased oxidative stress that then leads to increases in Abeta. Further, we propose that Abeta serves as a redox sensor and that oxidatively induced Abeta serves to attenuate oxidative stress. Obviously, whether Abeta is the culprit, as argued by the Amyloid Cascade Hypothesis, or a much maligned protector, as argued by the Alternate Amyloid Hypothesis, is clearly important to decipher to advance our understanding and design efficacious therapeutics for this disease. PMID- 16472208 TI - A partial failure of membrane protein turnover may cause Alzheimer's disease: a new hypothesis. AB - The amyloid hypothesis has dominated the thinking in our attempts to understand, diagnose and develop drugs for Alzheimer's disease (AD). This article presents a new hypothesis that takes into account the numerous familial AD (FAD) mutations in the amyloid precursor protein (APP) and its processing pathways, but suggests a new perspective beyond toxicity of forms of the amyloid beta-peptide (Abeta). Clearly, amyloid deposits are an invariable feature of AD. Moreover, although APP is normally processed to secreted and membrane-bound fragments, sAPPbeta and CTFbeta, by BACE, and the latter is subsequently processed by gamma-secretase to Abeta and CTFgamma, this pathway mostly yields Abeta of 40 residues, and increases in the levels of the amyloidogenic 42-residue Abeta (Abeta42) are seen in the majority of the mutations linked to the disease. The resulting theory is that the disease is caused by amyloid toxicity, which impairs memory and triggers deposition of the microtubule associated protein, Tau, as neurofibrillary tangles. Nevertheless, a few exceptional FAD mutations and the presence of large amounts of amyloid deposits in a group of cognitively normal elderly patients suggest that the disease process is more complex. Indeed, it has been hard to demonstrate the toxicity of Abeta42 and the actual target has been shifted to small oligomers of the peptide, named Abeta derived diffusible ligands (ADDLs). Our hypothesis is that the disease is more complex and caused by a failure of APP metabolism or clearance, which simultaneously affects several other membrane proteins. Thus, a traffic jam is created by failure of important pathways such as gamma-secretase processing of residual intramembrane domains released from the metabolism of multiple membrane proteins, which ultimately leads to a multiple system failure. In this theory, toxicity of Abeta42 will only contribute partially, if at all, to neurodegeneration in AD. More significantly, this theory would predict that focussing on specific reagents such as gamma-secretase inhibitors that hamper metabolism of APP, may initially show some beneficial effects on cognitive performance by elimination of acutely toxic ADDLs, but over the longer term may exacerbate the disease process by reducing membrane protein turnover. PMID- 16472210 TI - Natural and synthetic biologically active dimeric molecules: anticancer agents, anti-HIV agents, steroid derivatives and opioid antagonists. AB - Symmetry plays a crucial role in a variety of biological processes. For instance, many protein receptors, upon activation, dimerize to its active form and subsequently produce its biological action. Hence, there is a renewal of curiosity for the synthesis of dimeric molecules (or bivalent ligands) capable, not only to interact with specific biologic receptors, but also to induce greater biological responses than the corresponding monomeric counterpart. This is a vast and diverse theme of research. Hence, this review will discuss recent developments into this flourishing research field and will focus mainly into four specific topics namely dimeric 1) anticancer agents, 2) anti-HIV, 3) steroid derivatives and 4) opioid antagonists. PMID- 16472209 TI - 5-Lipoxygenase, leukotrienes biosynthesis and potential antileukotrienic agents. AB - Leukotrienes play an important role in the inflammatory process accompanying allergic diseases of respiratory, gastrointestinal and dermatological systems. Leukotrienes are generated from arachidonic acid as a result of the 5 lipoxygenase action. This paper deals with 5-lipoxygenase action mechanism and the following biosynthesis of all leukotrienes. In this article, potential antileukotrienic agents are classified according to their mechanism of action. The original antileukotrienic compounds of the Research Institute for Pharmacy and Biochemistry in Prague (VUFB), Czech Republic are presented in a separate chapter of the paper. PMID- 16472212 TI - Medicinal chemistry of N-acylhydrazones: new lead-compounds of analgesic, antiinflammatory and antithrombotic drugs. AB - In this article we provide an overview on the medicinal chemistry of new bioactive N-acylhydrazone (NAH) derivatives designed through the structural optimization of N-arylhydrazone precursors, originally planned by molecular hybridization of two known 5-lipoxygenase inhibitors, i.e. CBS-1108 and BW-755c. The analgesic, antiedematogenic and platelet anti-aggregating profile of several isosteric NAH compounds was investigated by using classical in vivo and ex-vivo pharmacological assays, which allowed the identification of new potent centrally and peripherically-acting analgesic leads, new antiinflammatory agents and new antithrombotic prototypes. During this study, dozens of active NAH compounds were discovered, clarifying the structure-activity relationships for this series of derivatives and indicating the pharmacophoric character of the N-acylhydrazone moiety for its biological profile. PMID- 16472211 TI - Allosteric regulators of the proteasome: potential drugs and a novel approach for drug design. AB - The proteasome recently gained an exceptional attention as a novel drug target, therefore its inhibitors became important subjects for rational drug design. A synthetic competitive inhibitor Velcade was lately approved in a fast-track process to treat multiple myeloma and is tested with other types of cancers. The proteasome is a major proteolytic assembly in eukaryotic cells responsible for the degradation of most intracellular proteins, including proteins crucial to cell cycle regulation and apoptosis. The ubiquitin-proteasome pathway has been implicated in many diseases such as cancer, autoimmune diseases, inflammation, and stroke. The activity of the proteasome can be blocked for therapeutic purposes with competitive inhibitors like Velcade, which trigger apoptosis in target cells. However, much more versatile outcomes and a true control of the proteasome can be achieved with allosteric regulators. Certain natural proteins and peptides bind to the catalytic core of the proteasome and allosterically induce a wide array of effects ranging from changes in product size to substrate specific inhibition. Designing small synthetic compounds allosterically interacting with the proteasome represents a novel approach that has enormous potential for the treatment of a wide range of diseases. Below we provide a review of current knowledge about proteasomal allosteric ligands. PMID- 16472213 TI - Chemistry and biological activity of natural and synthetic prenyloxycoumarins. AB - Prenyloxycoumarins and prenyloxyfuranocoumarins (isopentenyloxy-, geranyloxy-, linear and cyclic sesquiterpenyloxy compounds and their biosynthetic derivatives) represent a family of secondary metabolites that have been considered for years just as intermediates of other coumarin-based compounds. Only in the last two decades these secondary metabolites have been recognized as interesting and valuable biologically active natural products. Up to now more than 160 compounds have been isolated from plants mainly belonging to the families of Rutaceae and Umbelliferae, comprising common edible vegetables and fruits like lemons, oranges and grapefruits. In view of the biological activity of some natural prenyloxycoumarins, very recently syntheses of structurally related analogs aimed to establish detailed structure-activity relationships have also been carried out. Many of the isolated prenyloxy- and prenyloxy-furanocoumarins and their semisynthetic derivatives were shown to exert in vitro and in vivo remarkable anti-tumoral, anti-inflammatory and anti-viral effects. The object of this review is to examine in detail the different types of prenyloxycoumarins and prenyloxyfuranocoumarins from the chemical, phytochemical and biological point of view. PMID- 16472214 TI - In silico approaches to prediction of aqueous and DMSO solubility of drug-like compounds: trends, problems and solutions. AB - The solubility of drugs and drug-like compounds has been the subject of extensive studies aimed at finding a way to predict solubility from molecular structure. The aqueous solubility of a drug is an important factor that influences its absorption, distribution and elimination in the body. Poor aqueous solubility often causes a drug to appear inactive and may cause other biological problems. Compound solubility in DMSO represents another serious problem in early stages of drug discovery. An appreciation of the factors affecting a compound's DMSO solubility could help in predicting the storage conditions and appropriateness of compounds for primary bioscreening programs. In silico procedures for estimation of water and DMSO solubility represent extremely useful tools for the drug discovery practitioners. In this review, we provide a critical discussion of in silico models for the prediction of DMSO and water solubility of drug-like compounds used for virtual screening. We describe the main tendencies in the field, "booming" approaches and unsolved problems. A critical analysis of the accuracy and applicability of methods is provided. PMID- 16472215 TI - Function oriented synthesis: the design, synthesis, PKC binding and translocation activity of a new bryostatin analog. AB - Bryostatin 1 represents a novel and potent therapeutic lead with a unique activity profile. Its natural and synthetic availability is severely limited. Function oriented synthesis provides a means to address this supply problem through the design of synthetically more accessible simplified structures that at the same time incorporate improved functional activity. Pharmacophore searching and a new computer aided visualization of a possible binding mode are combined with an understanding of function and knowledge of synthesis to design and prepare a new and simplified compound with bryostatin-like function in biological systems. This new compound is a potent ligand for protein kinase C in vitro (K(i) = 8.0 nM). More significantly, the described molecule retains the functional ability to translocate a PKCdelta-GFP fusion protein in RBL cells. The extent of protein translocation and the sub-cellular localization induced by this new compound is similar to that seen in response to bryostatin 1, indicating that the new molecule retains the functional activity of the natural product but is simpler and can be synthesized in a practical fashion. PMID- 16472216 TI - Genetically engineered mouse models for drug discovery: new chemical genetic approaches. AB - While standard transgenic and knockout mouse technologies have provided a wealth of information for target selection and validation, there have been great advances in using more sophisticated modeling techniques to achieve temporal and spatial regulation of individual genes in adult animals. Recent developments in RNA interference (RNAi) technology in in vivo models promise to further improve upon the static and irreversible features of gene knockouts. Chemical genetic approaches create novel functional alleles of targets and allow fine modulation of protein function in vivo by small molecules, providing the most pharmacologically relevant target validation. Using these advanced models, one can not only ask whether the function of the target is critical for the initiation and maintenance of the disease, but also whether therapies designed to alter the function of the target would be safe and efficacious. In this review, we describe various in vivo tools for target validation in mouse models, discuss advantages and disadvantages of each approach, and give examples of their impact on drug discovery. PMID- 16472217 TI - Experiences in implementing uHTS--cutting edge technology meets the real world. AB - Driven by growing corporate compound files, the demands of target biology, and attempts to cut cost, the number of solutions to HTS has spiralled. In quick succession new assay technologies and screening platforms are appearing on the market, with the promise of screening faster than ever in low volume high density formats whilst providing high quality data. Within this world of rapid change, Pfizer has applied cutting edge technology to HTS by introducing screening in 1 microl formats utilising single molecule detection technology. Instead of resource intensive in-house development, Pfizer entered into a collaboration with Evotec OAI / Evotec Technologies and introduced their Mark-II EVOscreen platform. In this article we will outline the benefits of the approach taken at Pfizer, Sandwich, and introduce the Mark-II EVOscreen platform, illustrating the potential but also possible pitfalls of HTS miniaturisation. PMID- 16472218 TI - Systematic analysis of large screening sets in drug discovery. AB - Each year large pharmaceutical companies produce massive amounts of primary screening data for lead discovery. To make better use of the vast amount of information in pharmaceutical databases, companies have begun to scrutinize the lead generation stage to ensure that more and better qualified lead series enter the downstream optimization and development stages. This article describes computational techniques for end to end analysis of large drug discovery screening sets. The analysis proceeds in three stages: In stage 1 the initial screening set is filtered to remove compounds that are unsuitable as lead compounds. In stage 2 local structural neighborhoods around active compound classes are identified, including similar but inactive compounds. In stage 3 the structure-activity relationships within local structural neighborhoods are analyzed. These processes are illustrated by analyzing two large, publicly available databases. PMID- 16472219 TI - The FlexX database docking environment--rational extraction of receptor based pharmacophores. AB - We present an integrated docking environment that allows for iterative and interactive detailed analysis of many docking solutions. All docking information is stored in an ORACLE database. New scoring schemes (e.g. target-specific scoring functions) as well as various types of filters can be easily defined and tested within this environment. As an example application we investigated the validity of the following hypothesis: If a docking procedure can lead to enrichments significantly better than random then a bias towards (partially) correct placements should be detectable. Such bias in terms of a preference for certain interacting groups within the active site can be used to select a set of receptor-based pharmacophore constraints, which in turn might be used to enhance the docking procedure. As a proof of concept for this approach we performed docking studies on three targets: thrombin, the cyclin-dependent kinase 2 (CDK2) and the angiotensin converting enzyme (ACE). We docked a set of known active compounds with standard FlexX and derived three sets of target-specific receptor based pharmacophore constraints by statistical analysis of the predicted placements. Applying these receptor-based constraints in a virtual screening protocol utilizing FlexX-Pharm led to significantly improved enrichments. PMID- 16472220 TI - Assessment of the health effects of chemicals in humans: I. QSAR estimation of the maximum recommended therapeutic dose (MRTD) and no effect level (NOEL) of organic chemicals based on clinical trial data. AB - The primary objective of this investigation was to develop a QSAR model to estimate the no effect level (NOEL) of chemicals in humans using data derived from pharmaceutical clinical trials and the MCASE software program. We believe that a NOEL model derived from human data provides a more specific estimate of the toxic dose threshold of chemicals in humans compared to current risk assessment models which extrapolate from animals to humans employing multiple uncertainty safety factors. A database of the maximum recommended therapeutic dose (MRTD) of marketed pharmaceuticals was compiled. Chemicals with low MRTDs were classified as high-toxicity compounds; chemicals with high MRTDs were classified as low-toxicity compounds. Two separate training data sets were constructed to identify specific structural alerts associated with high and low toxicity chemicals. A total of 134 decision alerts correlated with toxicity in humans were identified from 1309 training data set chemicals. An internal validation experiment showed that predictions for high- and low-toxicity chemicals were good (positive predictivity >92%) and differences between experimental and predicted MRTDs were small (0.27-0.70 log-fold). Furthermore, the model exhibited good coverage (89.9-93.6%) for three classes of chemicals (pharmaceuticals, direct food additives, and food contact substances). An additional investigation demonstrated that the maximum tolerated dose (MTD) of chemicals in rodents was poorly correlated with MRTD values in humans (R2 = 0.2005, n = 326). Finally, this report discusses experimental factors which influence the accuracy of test chemical predictions, potential applications of the model, and the advantages of this model over those that rely only on results of animal toxicology studies. PMID- 16472221 TI - Development of immunopharmacotherapy against drugs of abuse. AB - Drug addiction is a major worldwide medical and social problem that continues to escalate. The addiction syndrome is remarkably similar between different drugs of abuse, and can be characterized as a chronic relapsing brain disorder with neurobiological changes that lead to a compulsion to take a drug with loss of control over drug intake. Presently used medications for the treatment of dependence disorders are based on drugs that are either agonists or antagonists of drugs of abuse, and have yielded only limited success. Immunopharmacotherapy is based on the generation or administration of antibodies that are capable of binding the targeted drug before it can reach the brain, whereas replacement strategies based on agonists or antagonists of these drugs generally cause many undesired side effects. A large amount of data has been gathered in recent years on the effects of active and passive immunization against cocaine, nicotine, PCP and methamphetamine in animal models, suggesting potential efficacy of these treatments in humans; and clinical trials are currently underway for vaccines against cocaine and nicotine. PMID- 16472222 TI - Ion channel drug discovery and research: the automated Nano-Patch-Clamp technology. AB - Unlike the genomics revolution, which was largely enabled by a single technological advance (high throughput sequencing), rapid advancement in proteomics will require a broader effort to increase the throughput of a number of key tools for functional analysis of different types of proteins. In the case of ion channels -a class of (membrane) proteins of great physiological importance and potential as drug targets- the lack of adequate assay technologies is felt particularly strongly. The available, indirect, high throughput screening methods for ion channels clearly generate insufficient information. The best technology to study ion channel function and screen for compound interaction is the patch clamp technique, but patch clamping suffers from low throughput, which is not acceptable for drug screening. A first step towards a solution is presented here. The nano patch clamp technology, which is based on a planar, microstructured glass chip, enables automatic whole cell patch clamp measurements. The Port-a Patch is an automated electrophysiology workstation, which uses planar patch clamp chips. This approach enables high quality and high content ion channel and compound evaluation on a one-cell-at-a-time basis. The presented automation of the patch process and its scalability to an array format are the prerequisites for any higher throughput electrophysiology instruments. PMID- 16472223 TI - The isolated perfused rat kidney model: a useful tool for drug discovery and development. AB - Over the past three decades, the Isolated Perfused Rat Kidney (IPK) has been used to study numerous aspects of renal drug disposition. Among the available ex-vivo methods to study renal transport, the IPK allows for elucidation of the overall contributions of renal transport mechanisms on drug excretion. Therefore, IPK studies can provide a bridge between in vitro findings and in vivo disposition. This review paper begins with a detailed overview of IPK methodology (system components, surgical procedure, study design). Various applications of the IPK are then presented. These applications include characterizing renal excretion mechanisms, screening for clinically significant drug interactions, studying renal drug metabolism, and correlating renal drug disposition with drug-induced changes in kidney function. Lastly, the role of IPK studies in drug development is discussed. Demonstrated correlations between IPK data and clinical outcomes make the IPK model a potentially useful tool for drug discovery and evaluation. PMID- 16472224 TI - Reversible covalent chemistry in drug delivery. AB - The targeting of drugs specifically to their sites of action is an important strategy for increasing drug efficacy. Chemists have come up with many elegant schemes that aim to convert drugs into magic bullets. This review focuses on the chemistry that underlies these schemes, with particular emphasis on two types of cleavable covalent bonds that are frequently used to link drugs to their various carriers: disulfide bonds and hydrazone bonds. These linkages have been used to release drugs under specific conditions; in the case of disulfides, cleavage is triggered by the mildly reducing environment found in intracellular fluids, and in the case of hydrazones, the acidic conditions that prevail in endosomes cause release of the drug. The applications of these chemistries in drug delivery are reviewed. PMID- 16472225 TI - Reverse pharmacognosy: application of selnergy, a new tool for lead discovery. The example of epsilon-viniferin. AB - The aim of reverse pharmacognosy is to find new biological targets for natural compounds by virtual or real screening and identify natural resources that contain the active molecules. To demonstrate the applicability of this concept, we report here a study on epsilon-viniferin, an active ingredient for cosmetic development. Nevertheless, this natural substance is weakly defined in terms of biological properties. SELNERGY, an inverse docking computer software, was used to identify putative binding biological targets for epsilon-viniferin. Among the 400 screened proteins two targets were retained. For cosmetic application, cyclic nucleotide phosphodiesterase 4 (PDE4) was the most interesting candidate. Moreover, other PDE subtypes (1, 2, 3, 5 and 6) were not retained, indicating a selectivity for PDE4. The experimental binding tests on the 6 subtypes of PDE revealed a significant selectivity of epsilon-viniferin for the PDE4 subtype. This selectivity was confirmed by evaluation of epsilon-viniferin on the secretion of TNF-alpha and Interleukin-8. Our data demonstrated that epsilon viniferin possesses anti-inflammatory properties by inhibiting PDE4 subtype. In conclusion, reverse pharmacognosy and its inverse docking component cannot only be integrated into a program for new lead discovery but is also a useful approach to find new applications for identified compounds. PMID- 16472226 TI - Macromolecular and nanotechnological modification of camptothecin and its analogs to improve the efficacy. AB - Camptothecin (CPT) and its analogs are some of the most potent antitumor agents known. However, their poor water-solubility and high toxicity require changes of their physicochemical and biological characteristics. Active lactone forms have provoked interest in the utility of CPT and its analogs again. Macromolecular chemical modifications and nanotechnological formulations have been used to obtain improved systems of CPT-related compounds. In these systems, one of the most important concepts is the enhanced permeability and retention (EPR) effect. The outcomes obtained by these approaches are displayed by introducing concrete examples. PMID- 16472227 TI - Application of the in combo screening approach for the discovery of non-alkaloid acetylcholinesterase inhibitors from Cichorium intybus. AB - Because of the direct correlation of cholinergic deficit and the severity of dementia, Alzheimer's disease is preferentially treated with acetylcholinesterase (AChE) inhibitors to supplement the acetylcholine level. In this study we focused on non-alkaloid AChE inhibitors from natural sources in order to discover new lead structures. In the course of in vitro extract screening of Tyrolean plants using an enzyme assay with Ellman's reagent, the dichloromethane extract of chicory roots (Cichorium intybus L.) showed a pronounced inhibitory effect on AChE. At a concentration of 1 mg extract/ml an inhibition of 70% was measured. Based on a 3D multi-conformational molecular-structure database consisting of secondary metabolites from C. intybus known from the relevant literature, virtual screening filtering experiments were conducted using both a feature-based pharmacophore model and a docking procedure. Some low molecular weight sesquiterpenoids exhibited distinct interactions with the pharmacophore model. In order to verify the applicability of this computer-aided strategy, an activity guided fractionation of the chicory root extract was performed, which resulted in the isolation of two sesquiterpene lactones, 8-deoxylactucin and lactucopicrin, showing significant and dose-dependent inhibitory activity on AChE (IC(50) of 308.1 microM [CI(95) 243.9 - 405.3 microM] and 150.3 microM [CI(95) 100.8 - 188.1 microM], respectively). The two isolates were correctly predicted within the virtual screening process which corroborates the potential of the computer assisted in combo screening approach for the discovery of the anti-cholinesterase compounds from C. intybus. PMID- 16472230 TI - QSAR modeling of carcinogenic risk using discriminant analysis and topological molecular descriptors. AB - A discriminant analysis model is presented for carcinogenic risk. The data set is obtained from the two-year rodent study FDA/CDER database and was divided into a training set of 1022 organic compounds and an external validation test set of 50 compounds. The model is designed to use as a decision support tool for a defined decision threshold, and is thus a binary discrimination into "high risk" and "low risk" categories. The carcinogenic risk classification is based on the method for estimating human risk from two-year rodent studies developed at the FDA/CDER/ICSAS. The paradigm chosen for this model allows a straightforward risk analysis based on historic information, as well as the computation of coverage, probability and confidence metrics that can further qualify the computed result. The molecular structures were represented as MDL mol files. The molecular structure information was obtained as topological structure descriptors, including atom-type and group-type E-State and hydrogen E-State indices, molecular connectivity chi indices, topological polarity, and counts of molecular features. The MDL QSAR software computed all these descriptors. Furthermore, the discriminant analyses were all performed with the MDL QSAR software. The reported model is based on fifty-three descriptors, using the nonparametric normal kernel method and the Mahalanobis distance to determine proximity. The model performed very well on the fifty compounds of the test set, yielding the following statistics: 76% correctly classified "high risk" (carcinogenic) and 84% correctly classified as "low risk" (non-carcinogenic). PMID- 16472228 TI - Artificial neural networks to optimize formulation components of a fixed-dose combination of rifampicin, isoniazid and pyrazinamide in a microemulsion. AB - The aim of this study to design a stable microemulsion formulation to deliver a combination of rifampicin, isoniazid and pyrazinamide in quantities suitable for administration to a paediatric population. The chemical stability of rifampicin, isoniazid and pyrazinamide alone and in various combinations was investigated in different solvents, solubilizing agents and surfactants. An artificial neural network was used to model data from the stability studies and a sensitivity analysis was applied to optimize the selection of the formulation components. Imwitor 308 and Crillet 3, exhibiting the highest overall positive sensitivity were selected to formulate the stable microemulsion. Due to drug dose specifications and solubility limitations, the final formulation contained only rifampicin and isoniazid, since the solubility of pyrazinamide in the lipid and aqueous components of the microemulsion did not achieve the required dose. The stability and solubility of rifampicin were improved in the formulation. Solubilization of the rifampicin in the lipid droplets of the internal phase and lipophilic chains of the surfactants increased the quantity of rifampicin that can be incorporated, while protecting it from oxidative degradation and also limited its contact with isoniazid, which has been shown to affect its stability. The results of this study indicate that the Artificial Neural Network can be successfully used to optimize the choice of solvents, solubilizing agents and surfactants prior to formulation of the microemulsion, limiting the amount of experiments, thus reducing the costs during the preformulation study. PMID- 16472231 TI - Exploration tools for drug discovery and beyond: applying SciFinder to interdisciplinary research. AB - Chemists have long recognized the value of online databases for surveying the literature of their field. Chemical Abstracts Service (CAS) databases covering almost a century's worth of journal articles and patent documents are among the best known and widely used for searching information on compounds. Today's research presents a new challenge, however, as the boundaries of chemistry and biological sciences overlap increasingly. This trend is especially true in the drug discovery field where published findings relating to both chemical and biological entities and their interactions are examined. CAS has expanded its resources to meet the requirements of the new, interdisciplinary challenges faced by today's researchers. This is evident both in the content of CAS databases, which have been expanded to include more biology-related information, and in the technology of the search tools now available to researchers on their desktop. It is the integration of content and search-and-retrieval technology that enables new insights to be made in the vast body of accumulated information. CAS's SciFinder is a widely used research tool for this purpose. PMID- 16472232 TI - Challenges of target/compound data integration from disease to chemistry: a case study of dihydrofolate reductase inhibitors. AB - Despite the improvements in informatics associated with initiatives in the structure-based design and genomics fields, no straight-forward links are available between a given disease class and drug chemistry. This involves effective linking of disease to protein targets, and then mapping these targets to drug chemistry. In practice, protein-ligand structural analyses and high throughput screening experiments generate the links between targets implicated in disease and chemical leads. Additionally, large volumes of relevant data are also being produced by high-throughput X-ray crystallography and in-silico docking initiatives. Each of these efforts takes a distinctly different approach to how data is managed and mined, resulting in difficulties in sharing data across each area. This review discusses the diverse approaches taken to data management in these areas, and the challenges associated with the construction of a data warehouse that meets all of the needs of each data type. Using the current work available for dihydrofolate reductase inhibitors, we demonstrate the challenges and opportunities associated with data mining from disease to drug chemistry. PMID- 16472233 TI - New approaches to mechanism analysis for drug discovery using DNA microarray data combined with KeyMolnet. AB - We have developed a comprehensive information platform, named KeyMolnet, for drug discovery and life science research in the post-genome era. Using KeyMolnet, we show new approaches to research into the biological mechanism in DNA microarray analysis. Thanks to the DNA microarray technology, it is now possible to obtain very large quantities of gene expression data at a time. However, it is still difficult to extract meaningful information from such large quantities of data and to analyze the relationship between gene expression data and biological function. We therefore developed an advanced tool that can generate molecular networks upon demand, and beyond signaling "cross-talks," can connect them to physiological phenomena and medical and drug information. Here we show the methods of mechanism analysis using the DNA microarray data and KeyMolnet, as well as the possible mechanism of inducing apoptosis in the human promyelocytic leukemia cell line, HL-60, treated with 12-O-tetradecanoylphorbol 13-acetate (TPA), using the time series of gene expression data from DNA microarray experiments. KeyMolnet enables practical approaches to research into biological mechanisms, which in turn contribute to new discoveries in the medical, pharmaceutical and life sciences. PMID- 16472234 TI - Comprehensive computational assessment of ADME properties using mapping techniques. AB - One strategy to potentially improve the success of drug discovery is to apply computational approaches early in the process to select molecules and scaffolds with ideal binding and physicochemical properties. Numerous algorithms and different molecular descriptors have been used for modeling ligand-protein interactions as well as absorption, distribution, metabolism and excretion (ADME) properties. In most cases a single data set has been evaluated with one approach or multiple algorithms that have been compared for a single dataset. These models have been primarily evaluated by leave-one out analysis or boot strapping with groups representing 25-50% of the training set left out of the final model. In a very few examples a test set of molecules not included in the model has been used for an external evaluation. In the present study we have applied Sammon non linear maps, Support Vector Machines and Kohonen Self Organizing Maps to modeling numerous datasets for ADME properties including human intestinal absorption, blood brain barrier permeability, cytochrome P450 binding, plasma protein binding, P-gp inhibition, volume of distribution and plasma half life. PMID- 16472235 TI - Predicting dopamine receptors binding affinity of N-[4-(4-Arylpiperazin-1-yl) butyl]aryl carboxamides: computational approach using topological descriptors. AB - Relationship between the topological indices and Dopamine D3 and D4 receptor binding affinities of N-[4-(4-Arylpiperazin-1-yl)butyl]aryl carboxamides has been investigated. Three topological indices, the Wiener's Index- a distance-based topological descriptor, molecular connectivity index- an adjacency based topological descriptor and eccentric connectivity index- an adjacency-cum distance based topological descriptor were used for the present investigations. A data set comprising of 37 substituted N-[4-(4-Arylpiperazin-1-yl)butyl]aryl carboxamides was selected for the present studies. The values of the Wiener's index, eccentric connectivity index and molecular connectivity index for each of the 37 analogues comprising the data set were computed using in-house computer program. Resultant data was subsequently analyzed and suitable models were developed after identification of active ranges. Subsequently, a biological activity was assigned to each analogue using these models, which was then compared with the reported D3 and D4 receptor binding affinity. These models exhibited exceptionally high predictability. PMID- 16472236 TI - Combinatorial chemistry and fragment screening--two unlike siblings? AB - Efficiently scouting the chemical space is one of the major challenges for lead discovery for drug development. In recent times some shifts have been made away from HTS and combinatorial chemistry to more focused approaches. Combinatorial chemistry was the starting point for the development of synthesis concepts that were intended to cover and explore the chemical space without having to prepare every individual compound. In this review, these lead finding approaches will be discussed comparing virtual and synthesized libraries. In addition we discuss the concepts and relationships of evolutionary libraries using genetic algorithms and dynamic combinatorial chemistries, as well as templated fragment ligation concepts. Taking a more abstract view of all approaches, the concepts may loop back into Combinatorial Chemistry allowing a more educated choice of building blocks and chemistries. PMID- 16472237 TI - Autocorrelation of molecular electrostatic potential surface properties combined with partial least squares analysis as alternative attractive tool to generate ligand-based 3D-QSARs. AB - A database of 106 human A3 adenosine receptor antagonists was used to derive two alternative PLS models: one starting from CoMFA descriptors and the other starting from the autocorrelation descriptors. The peculiarity of this work is the introduction of autocorrelation vectors as molecular descriptors for the PLS analysis. The autocorrelation allows comparing molecules (and their properties) with different structures and with different spatial orientation without any previous alignment. In particular, Molecular Electrostatic Potential (MEP) was the property computed and its information encoded in autocorrelation vectors. The 3D spatial distribution and the values of the electrostatic potential is in fact largely responsible for the binding of a substrate to its receptor binding site. Validation was done with an external test set and the results of the two models were compared. Interestingly, our preliminary results seem to indicate that this new alternative approach could robustly compete with the already well consolidated CoMFA approach. In particular, we have suggested that it could be a very interesting tool to filter large structural database in several virtual screening applications. PMID- 16472238 TI - Preparation of cherry-picked combinatorial libraries by string synthesis. AB - String synthesis [1-3] is an efficient and cheap manual method for preparation of combinatorial libraries by using macroscopic solid support units. Sorting the units between two synthetic steps is an important operation of the procedure. The software developed to guide sorting can be used only when complete combinatorial libraries are prepared. Since very often only selected components of the full libraries are needed, new software was constructed that guides sorting in preparation of non-complete combinatorial libraries. Application of the software is described in details. PMID- 16472240 TI - The characterisation of the collagenolytic activity of cardosin a demonstrates its potential application for extracellular matrix degradative processes. AB - Type I collagen is the major fibrous protein of mammals being needed to strengthen and organise the extracellular matrix (ECM). Connective tissue components are modulated by matrix metalloproteinases, which are critical for disintegration and remodelling of ECM under physiological and pathological conditions. Cardosin A is an abundant aspartic proteinase (AP) from Cynara cardunculus L. that has been shown to be able to hydrolyse fibrillar collagen within the alpha-chains. The aim of this work is the characterisation of collagen degradation by cardosin A, since in the native state fibrillar collagen is resistant to most proteolytic enzymes. The pattern of type I collagen hydrolysis by cardosin A is defined and maintained for at least 24 hours of digestion, suggesting that cardosin A can hydrolyse collagen at a small number of specific peptide bonds. N-terminal sequencing of hydrolysis products identified one cleavage site as being Phe464-Gln465 in the alpha2 chain of collagen I. Two peptides were synthesised correspondent to collagen I specific sequences, in order to produce two polyclonal antibodies, that allowed the identification of three collagen fragments following cardosin A cleavage. Defining the mechanism of collagen cleavage by collagenases and other enzymes, like cardosin A, is important for the comprehension of physiological and pathological processes affecting the ECM. To our knowledge, this is the first study of in vitro collagenolytic activity of a plant AP. Therefore, in view of the cardosin A restricted specificity towards collagen this enzyme may be proposed for an eventual medical or technical procedures assisting ECM remodelling. PMID- 16472241 TI - Assessment of the health effects of chemicals in humans: II. Construction of an adverse effects database for QSAR modeling. AB - The FDA's Spontaneous Reporting System (SRS) database contains over 1.5 million adverse drug reaction (ADR) reports for 8620 drugs/biologics that are listed for 1191 Coding Symbols for Thesaurus of Adverse Reaction (COSTAR) terms of adverse effects. We have linked the trade names of the drugs to 1861 generic names and retrieved molecular structures for each chemical to obtain a set of 1515 organic chemicals that are suitable for modeling with commercially available QSAR software packages. ADR report data for 631 of these compounds were extracted and pooled for the first five years that each drug was marketed. Patient exposure was estimated during this period using pharmaceutical shipping units obtained from IMS Health. Significant drug effects were identified using a Reporting Index (RI), where RI = (# ADR reports / # shipping units) x 1,000,000. MCASE/MC4PC software was used to identify the optimal conditions for defining a significant adverse effect finding. Results suggest that a significant effect in our database is characterized by > or = 4 ADR reports and > or = 20,000 shipping units during five years of marketing, and an RI > or = 4.0. Furthermore, for a test chemical to be evaluated as active it must contain a statistically significant molecular structural alert, called a decision alert, in two or more toxicologically related endpoints. We also report the use of a composite module, which pools observations from two or more toxicologically related COSTAR term endpoints to provide signal enhancement for detecting adverse effects. PMID- 16472242 TI - Present status and future prospects of chemotherapeutics for intractable infections due to Mycobacterium avium complex. AB - Mycobacterium avium-intracellulare complex (MAC) infections are frequently encountered in immunocompromised hosts, especially AIDS patients, although nodular-bronchiectasis type MAC infections without predisposing conditions are steadily increasing, particularly in Japan. Clinical control of MAC infection is difficult, since it responds poorly to available antimycobacterial regimens because of the intrinsic resistance of MAC organisms to common antituberculosis drugs, although some antimycobacterial drugs, such as macrolides (clarithromycin, azithromycin), ethambutol, clofazimine and rifamycins (especially rifabutin), are fairly or modestly effective in controlling AIDS-associated MAC bacteremia. In addition, treatment of pulmonary MAC infections is also difficult, even with available antimycobacterial multi-drug therapies. The insufficient efficacy of the majority of ordinary antimycobacterial drugs in treating MAC diseases is principally due to the low susceptibility of MAC to the majority of ordinary antimycobacterial drugs, the extremely wide range of susceptibilities of MAC isolates to most antimicrobial drugs, and the fact that polyclonal MAC infections are occasionally seen in AIDS patients. Therefore, the development of new antimicrobials and administration protocols that are safe and potently effective against MAC infections is urgently needed. However, despite the gradual but steady increase in the incidence of MAC infections, and thereby the urgent call for new drug development, new drugs that are truly useful for the treatment of refractory MAC diseases continue to elude us. In this review article, the following topics will be described: (1) the present status of the existing anti MAC drugs, with special reference to the recent findings of the in vitro and in vivo anti-MAC activities of macrolides and other moderately useful drugs, such as new rifamycins, clofazimine and some fluoroquinolones, and (2) the present status and future prospects of the development of new promising antimicrobials with anti MAC activity. PMID- 16472239 TI - Protective effect of urate oxidase on uric acid induced-monocyte apoptosis. AB - Uremic patients have a higher risk of infection and malignancy than normal subjects. Previous studies have deomonstrated that monocytes isolated from uremic patients display an increased apoptosis rate compared to normal subjects; furthermore uremic plasma can increase apoptosis rates on U937, a human monocytic cell line. In several pathological conditions, precipitation of uric acid crystals can lead to renal insufficiency or acute renal failure by different mechanisms. In recent studies uric acid has been shown to induce inflammatory response from monocytes and it has been suggested to be involved in cell dysfunction. Rasburicase is a new recombinant urate oxidase developed to prevent and treat hyperuricaemia in patients with cancer or renal failure; it degrades uric acid to allantoin, a less toxic and more soluble product. In the present study, we aimed at determining whether uric acid may be a factor affecting U937 apoptosis, and whether urate oxidase may reduces or even prevent uric acid induced cell apoptosis. Hoechst staining and internucleosome ledder fragmentation of DNA showed that uric acid increased the percentage of apoptotic cells comparing to the control and that when the U937 cells were incubated with uric acid and urate oxidase the percentage of apoptosis significantly decreased (from 43+/-7% to 19+/- 3%, p<0.05). Also, the activity of caspase-8 and caspase-3 showed the same trend (caspase 3: from 2.7+/-0.53 to 1.6+/-0.42; caspase-8: from 2.2+/-0.43 to 1.3+/-0.57). A reduction of intracellular reduced glutathione (GSH) concentration was found in uric acid treated cells while the addition of urate oxidase in the uric acid incubated cells decreased the GSH extrusion. The concentration of TNF-alpha was increased in the sample incubated with uric acid comparing to the control. Uric acid is an inducer of apoptosis on U937 cell line, and therefore it may be a component of the mosaic of uremic toxins both in acute and chronic renal disease. We can hypothesize that uric acid might be directly involved in the apoptotic process trough the activation of both death receptor and mitochondrial-mediated pathways. We have, also, demonstrated that urate oxidase is able to prevent at least in part, the effect of uric acid on U937 apoptosis. This effect might be a result of different mechanisms of action. PMID- 16472243 TI - Microdialysis in drug discovery. AB - Microdialysis has been developed during the last 25 years by several authors primarily to study brain function and changes in levels of endogenous compounds such as neurotransmitters or metabolites. The development of microdialysis for the purpose of measuring drugs was initiated during the late eighties. This technique provides a means of continuous plasma sampling without repeated blood sampling and the applicability to the study of drug metabolism and pharmacokinetics in experimental animals and human. Also, the microdialysis technique allows the study of plasma protein binding and the saturation of protein binding. The implantation of the microdialysis probe in other tissues and organs, like central nervous system, adipose tissue and heart, allows the study of drug distribution. On the other hand, the measurement of endogenous substances using the microdialysis technique permits the study of the effect of drugs on neurotransmission and metabolism. Moreover, as this technique allows the simultaneous determination of different physiological parameters such as blood pressure, locomotor and convulsive activity, it is a suitable tool for pharmacokinetic-pharmacodynamic studies of drugs and pharmacokinetic pharmacodynamic (PK-PD) modeling. Lastly, the reverse microdialysis is a powerful technique for the study of local actions of drugs in different tissues such as specific brain nuclei, myocardium, liver or skeletal muscle. So, this article reviewed the vast applications of the microdialysis technique for the study of pharmacokinetic and pharmacodynamic properties of drugs. PMID- 16472244 TI - The role of atelocollagen-based cell transfection array in high-throughput screening of gene functions and in drug discovery. AB - The human genome project has been completed, but the function of many genes is unknown. It is, therefore, necessary to elucidate the function of a large number of genes within a short time. To achieve this goal, materials are needed that condense or package DNA into nano-particles that can easily be taken up by cells and would allow DNA to be retained without degradation. Atelocollagen is a reliable carrier for gene delivery because it is considered safe and appropriate for practical use. We developed a basic technique for high-throughput gene transfer and expression screening by pre-coating a multi-well plate with an Atelocollagen/DNA complex in which cells are then seeded. Complexes with a nano particle form were efficiently transduced into cells without the use of additional transfection reagents, and they allowed for long-term gene expression. The complex spotted onto the well of a plate was stable for a long period and allowed the cells to transduce and express reporter genes. We also showed that the present method with Atelocollagen-based gene transfer is applicable to gene medicines, such as antisense ODNs, siRNA, and adenovirus vectors. These results suggest that an Atelocollagen-based cell transfection array may be appropriate for general use in the high-throughput screening of large sets of gene medicines with functions in mammalian cells. PMID- 16472245 TI - Mass spectrometry in drug discovery: a current review. AB - Drug discovery in the pharmaceutical industry has shown great demands for screening absorption, distribution, metabolism, excretion (ADME) and pharmacokinetics (PK) in guiding the selection of lead candidate compounds. Determination of ADME/PK properties of new chemical entities (NCE) in early drug discovery should allow defects to be corrected prior to time-consuming and expensive preclinical and clinical development stages. Mass spectrometry has evolved to become an irreplaceable technology in all types of drug discovery applications because of its high sensitivity, speed, selectivity, versatility, and ease of automation. This review will include current mass spectrometric techniques and applications in drug discovery, as well as future prospects. PMID- 16472247 TI - Efficient optimization strategy for marginal hits active against abl tyrosine kinases. AB - Primary high-throughput screening of commercially available small molecules collections often results in hit compounds with unfavorable ADME/Tox properties and low IP potential. These issues are addressed empirically at follow-up lead development and optimization stages. In this work, we describe a rational approach to the optimization of hit compounds discovered during screening of a kinase focused library against abl tyrosine kinase. The optimization strategy involved application of modern chemoinformatics techniques, such as automatic bioisosteric transformation of the initial hits, efficient solution-phase combinatorial synthesis, and advanced methods of knowledge-based libraries design. PMID- 16472248 TI - Strategies for compound selection. AB - In-house pharmaceutical collections are no longer sufficient for sampling chemical spaces. As novel bioactive chemotypes are successfully identified by virtual and high -throughput screening, the ability to rapidly sift through large numbers of chemicals prior to acquisition or experiment is required. Strategies for compound selection include some of the following steps: 1.) database assembly ('in silico' inventory); 2a.) structural integrity verification (keep unique structures only); 2b.) limited exploration of alternative chemical representations for the uniques (stereoisomers, tautomers, ionization states); 3.) property and structural filtering (remove unwanted structures); 4.) 3D structure generation (for virtual screening or 3D-based similarity); 5a.) clustering or statistical design for selection; 5b.) similarity-based selection (if bioactives are known); 5c.) receptor-based selection (if target binding site is known); 6.) add a random subset to the final list. PMID- 16472249 TI - Prediction of glomerular filtration rate from body cell mass and plasma creatinine. AB - The gold standards for the measurement of glomerular filtration rate (GFR) are inulin clearance and radioisotopic methods. However, creatinine clearance is the most used test to evaluate GFR in clinical practice. Its adequacy is questionable, since its repeatability is quite poor, mainly due to errors in the collection of urine. The aim of this study was to evaluate a new method to predict GFR from the body cell mass (BCM) and plasma creatinine (Pcr), avoiding urine collection. The values of BCM were obtained in 275 adult renal patients with different renal function, ranging from normality to advanced renal failure. The relationship of GFR (clearance of (99m)Tc-DTPA) with BCM and Pcr was calculated in the first 85 patients. A highly significant linear correlation was found between GFR and the ratio BCM/Pcr. Thereafter, GFR was predicted from BCM and Pcr (BCM GFR) with formulas derived from the relationships found between GFR and the ratio BCM/Pcr. For comparison, GFR was predicted also according to other prediction formulas: Cockcroft and Gault (CG GFR), and the simplified MDRD formula (MDRD GFR). BCM GFR gave a more precise estimate of GFR than CG GFR and MDRD GFR. In fact, BCM GFR had the best correlation and agreement with true GFR ((99m)Tc-DTPA). Furthermore, CG GFR and MDRD GFR markedly overestimated true GFR. Finally, the error of prediction of BCM GFR was definitely lower than that of the two other estimates of GFR. GFR can be predicted from BCM and plasma creatinine. This method, which is very simple and accurate, seems suitable to establish the adequate dosage of drugs cleared by the kidneys. PMID- 16472250 TI - Applications of multi-angle laser light-scattering detection in the analysis of peptides and proteins. AB - The proliferation of new peptides and proteins requiring characterisation is a direct result of recent advances in genomics and proteomics, but protein aggregation is particular problem in the biotechnology industry, where aggregation is encountered throughout the lifetime of a therapeutic protein, including during refolding, purification, sterilization, shipping, and storage process. To ensure that it meets quality standards, the size, molecular weight and/or molecular weight distribution, and aggregate state must be accurately determined. Traditional analytical methods for determining molecular weight include size-exclusion chromatography (SEC), gel electrophoresis, analytical ultracentrifugation and time-of-flight mass spectrometry. These technologies are time-consuming (some take days), provide data based on relative standards, or cannot characterise very high molecular weight aggregates. Laser light-scattering (LS) detection coupled with SEC system have been used for over a decade to determine the size and molecular weight of bio-molecules such as proteins, peptides, polysaccharides, oligonucleotides, and antibodies, the method of choice being for molar mass determinations and the study of self-association and heterogeneous interaction under native, equilibrium conditions in solution. The purpose of the current review is to describe and discuss the capability of the SEC/LS system to determine absolute molecular weight of proteins and their complexes and the association state of the conjugate, either with itself or with protein receptor/ligands. For this, the "two or three detector" methods, each with its advantages and limitations, can be used to calculate the molecular weight of a simple protein or glycoprotein, and the stoichiometry of their complexes. Also, some alternative techniques for determining the molecular weight are discussed in this review. Applications of all these methodologies are described. PMID- 16472251 TI - Is phage display technology on target for developing peptide-based cancer drugs? AB - New tumor targeting agents are required to advance cancer diagnosis and treatment. Bacteriophage (phage) display technology, a molecular genetic means of combinatorial drug discovery, is an emerging approach to identify and improve peptide molecules as pharmaceuticals. Peptides are thought to have clinically desirable benefits over currently used biomolecules, such as antibodies, because of their rapid blood clearance, increased diffusion and tissue penetration, non immunogenic nature and ease of synthesis. Using phage display, one can rapidly and simultaneously survey billion-clone peptide libraries, resulting in large numbers of "hits". However, only a few lead compounds resulting from the hits historically reach the drug market. Hence determining which peptide may best translate into a useful drug is of particular importance. Examination of successfully marketed drugs has highlighted key features of a winning agent, including low molecular weight, high affinity, stability, solubility, lipophilicity and conformational rigidity. Although peptide modulators of tumor cell function and cancer targeting agents have been developed, the majority of peptide-based drugs reported thus far are immune and cardiac regulators. In this review, we will highlight how phage display has been employed to isolate peptides that target key steps in cancer progression--from tumor growth to metastasis--and how phage display technology can be harnessed to select a priori peptides with inherent features essential for anti-cancer drug efficacy. In 2003, phage display provided us with several novel peptides not only in clinical trials but approved by the FDA for use as therapeutics in a variety of diseases--suggesting that the future looks bright for phage display in anti-cancer drug development. PMID- 16472252 TI - Probing protein binding sites by circular dichroism spectroscopy. AB - Pharmacological and pharmacodynamic properties of biologically active natural and synthetic compounds are crucially determined via their binding to proteins of the human body. Several spectroscopic techniques are available to study these mainly non-covalent interactions. Circular dichroism (CD) spectroscopy, being sensitive to the chirality of ligand molecules induced by the asymmetric protein environment, has widely and successfully been applied for many decades. Chiral conformation of the ligand due to conformational adaptation to its binding site, or interaction between ligand molecules held in chiral arrangement relative to each other by the protein sites, results in one or more induced CD bands with different shape, sign and intensity. These extrinsic Cotton effects present in light absorbing region of the optically active or inactive ligand molecules give qualitative and quantitative information of the binding process. It can provide valuable data on the stereochemistry, number, location and nature of the binding sites. This paper is aimed to survey briefly the literature and the results of recent investigations undertaken in this field. PMID- 16472253 TI - Intramolecular hydrogen bonding in depsipeptides containing endo-3,6 tricyclo[6.2.1.0(2,7)]undeca-4,9-diene-3,6-endo-diol. AB - A variety of endo-3,6-tricyclo[6.2.1.0(2,7)]undeca-4,9-diene-3,6-endo-diol depsipeptide or bis-amino acid derivatives containing the same parallel strands were synthesised and examined for conformational preferences by NMR. The study indicates that this novel class of constrained peptides displays beta-turn-like and beta-sheet-like conformations. Evidences are supported by observation of the rates of proton-deuterium exchange, nuclear Overhauser effects and further by dynamic simulations, semi-empirical and ab initio calculations. The chemical shifts were measured in CDCl3 and CDCl3 with 15% DMSO-d6 solutions. It was carried out a van't Hoff analysis, which is in agreement with the theoretical studies. In chloroform-d, for the derivatives 3-6 the hydrogen-bonded species are enthalpically preferred but entropically disfavoured. PMID- 16472254 TI - Strategy for the development of a thermodynamically stable oral microemulsion. AB - Microemulsions, being thermodynamically stable systems, with low viscosity and elegant in appearance have attracted interest not only for the delivery of single drug substances with low water solubility but for the stabilization of drugs in combination due to their preferential solubility in either the water or oil phases. Microemulsion design involves the solubilisation of an optimum amount of the dispersed phase in the continuous phase, utilizing the minimum amount of surfactant/mixture of surfactants/cosurfactants. It is the choice of the surfactant/surfactant mixture and/or cosurfactants, which poses the greatest challenge in the design of a thermodynamically stable microemulsion formulation. This paper will present a strategy for choosing surfactants to achieve a stable, dilutable microemulsion formulation for oral administration. Ternary and pseudo ternary phase diagrams were constructed by titrating a series of mixtures [lipid (miglyol 812): cosurfactant and/or surfactant (cremophor RH, imwitor 308/742, sorbitol, brij 97, crillet 3)] with water at room temperature, the phases formed visually assessed after each addition and classified as isotropic (ME), liquid crystalline (LC) or coarse emulsions (EM). Results indicate that the surfactant combination of imwitor 308 and crillet 3 proved most successful in incorporating 25 to 30% miglyol 812 into a microemulsion formulation utilizing an intermediate quantity of surfactant and maintaining homogeneity on dilution. PMID- 16472255 TI - Rapid screening of antioxidants in pharmaceutical formulation development using cyclic voltammetry--potential and limitations. AB - Cyclic voltammetry (CV) is a unique technique for the electrochemical characterization of compounds by providing their oxidation/reduction potentials. This technique is widely used in evaluating antioxidants in the oil, food, diagnostic and agricultural industries; however, CV is rarely used in the development of pharmaceutical formulations. This review briefly describes the basic principles of CV and its application in other industries along with the potential and limitations of CV in the rapid evaluation of antioxidants in pharmaceutical formulations. An extensive survey of the literature shows that there is a good correlation between the oxidation potentials of various antioxidants and their antioxidant efficiency. In conclusion, CV should be useful in the development of pharmaceutical formulations where a small group of the preferred antioxidants is rapidly identified. This small group of the most preferred antioxidants can then be employed in a conventional drug stability study thereby providing a rapid approach for the selection of the most suitable antioxidant for a pharmaceutical formulation. PMID- 16472256 TI - The oxazolomycins: a structurally novel class of bioactive compounds. AB - Oxazolomycin, first isolated in 1985, is a novel bioactive compound, exhibiting potent antiviral, antibacterial and cytotoxic activity, and is now known to be the parent of a wider class of compounds. The broad-spectrum activity of oxazolomycin has been attributed to its protonophoric properties. This review outlines the isolation, structural determination, biosynthesis, bioactivity, biological mode of action and synthesis of all members of this compound class. Significantly, the oxazolomycins appear to offer not only a completely novel chemotype as a bioactive lead structure, but also a chemotype, which also exhibits an unusual biological mode of action. PMID- 16472259 TI - Monitoring medical research conduct: why, how and by whom? PMID- 16472260 TI - Pharmaceutical Benefits Scheme criteria for the use of tumour necrosis factor alpha inhibitors in the treatment of ankylosing spondylitis in Australia: are they evidence based? AB - BACKGROUND: In 2004, the Pharmaceutical Benefits Scheme (PBS) listed infliximab as a subsidised treatment for ankylosing spondylitis (AS). Eligibility to receive this treatment for AS involved fulfilling several criteria. AIM: To examine the medical literature concerning response to tumor necrosis factor (TNF)-alpha inhibitors in AS and compare with the PBS criteria for these agents. METHODS: Review of published studies and analysis of the PBS criteria for the prescription of TNF inhibitors for the treatment of AS to assess whether the published criteria are evidence based. RESULTS: The published findings on the prediction of response to TNF inhibitors in the treatment of AS suggest that age, duration of disease, disease activity, functional status at the time of commencement of TNF inhibitors and, possibly, level of acute phase reactants predict the outcome of treatment with TNF inhibitors in AS. The PBS criteria do not reflect the published findings on predictors of response to TNF inhibitors. CONCLUSION: The current PBS criteria that need to be fulfilled for patients to receive subsidised treatment with TNF inhibitors for AS are not evidence based and will lead to the selection of patients with established disease while excluding patients with early disease. PMID- 16472261 TI - Measurement of faecal fat excretion: an evaluation of attitudes and practices of Australian gastroenterologists. AB - BACKGROUND: Although faecal fat excretion over 72 h is the gold standard for quantifying fat malabsorption, there has been a push from chemical pathology laboratories to discontinue this test, arguing that it is unreliable and of limited clinical value. AIMS: To assess attitudes, knowledge and practices of Australian gastroenterologists in relation to the test and to gauge opinion as to whether it should remain available. METHODS: A self-administered questionnaire was developed to assess attitudes towards, patterns of use and understanding of 72-h faecal fat collections. This was posted to all members of the Gastroenterological Society of Australia. RESULTS: Of 429 eligible gastroenterologists, 124 (29%) responded. Eighty-two per cent utilized the test; 62% at least once per year. Main indications were suspected steatorrhoea (55%), unexplained chronic diarrhoea (39%) or weight loss (29%). Thirty-eight per cent ordered the test to determine stool volume. Only 26% attempted to appropriately fat load patients and approximately half did not recognize the potential influence of medications and stool volume. This was also reflected in poor interpretation of results in specific clinical scenarios. Of those who use the test at least once per year, 97% wanted its continued availability and 51% felt their practice would be significantly affected if the test was discontinued. CONCLUSIONS: Although continued availability of 72-h faecal fat estimation is supported by many Australian gastroenterologists, the test appears to be often performed and interpreted suboptimally. Rather than discard the test, efforts should more appropriately be directed to improving baseline knowledge to ensure its optimal performance and interpretation. PMID- 16472257 TI - Gi-coupled receptors mediate phosphorylation of CPI-17 and MLC20 via preferential activation of the PI3K/ILK pathway. AB - Sustained smooth-muscle contraction or its experimental counterpart, Ca2+ sensitization, by G(q/13)-coupled receptor agonists is mediated via RhoA dependent inhibition of MLC (myosin light chain) phosphatase and MLC20 (20 kDa regulatory light chain of myosin II) phosphorylation by a Ca2+-independent MLCK (MLC kinase). The present study identified the corresponding pathways initiated by G(i)-coupled receptors. Somatostatin acting via G(i)1-coupled sstr3 receptor, DPDPE ([D-Pen2,D-Pen5]enkephalin; where Pen is penicillamine) acting via G(i)2 coupled delta-opioid receptors, and cyclopentyl adenosine acting via G(i)3 coupled adenosine A1 receptors preferentially activated PI3K (phosphoinositide 3 kinase) and ILK (integrin-linked kinase), whereas ACh (acetylcholine) acting via G(i)3-coupled M2 receptors preferentially activated PI3K, Cdc42 (cell division cycle 42)/Rac1, PAK1 (p21-activated kinase 1) and p38 MAPK (mitogen-activated protein kinase). Only agonists that activated ILK induced sustained CPI-17 (protein kinase C potentiated inhibitor 17 kDa protein) phosphorylation at Thr38, MLC20 phosphorylation at Ser19, and contraction, consistent with recent evidence that ILK can act as a Ca2+-independent MLCK capable of phosphorylating the MLC phosphatase inhibitor, CPI-17, at Thr38. ILK activity, and CPI-17 and MLC20 phosphorylation were inhibited by LY294002 and in muscle cells expressing ILK(R211A) or treated with siRNA (small interfering RNA) for ILK. ACh acting via M2 receptors activated ILK, and induced CPI-17 and MLC20 phosphorylation and muscle contraction, but only after inhibition of p38 MAPK; all these responses were inhibited in cells expressing ILK(R211A). Conversely, ACh activated PAK1, a step upstream of p38 MAPK, whereas the three other agonists did so only in cells transfected with ILK(R211A) or siRNA for ILK. The results demonstrate reciprocal inhibition between two pathways downstream of PI3K, with ILK inhibiting PAK1, and p38 MAPK inhibiting ILK. Sustained contraction via G(i)-coupled receptors is dependent on CPI-17 and MLC20 phosphorylation by ILK. PMID- 16472262 TI - Communication deficiencies in research and monitoring by ethics committees. AB - AIM: To assess the range of issues that arise for researchers, research participants and ethics committee members in the setting of a hospital-based research environment and to develop a tool that could be used to assist in the process of monitoring. METHODS: A qualitative phase comprising focus group sessions and interviews involving research participants, researchers and ethics committee members of a public teaching hospital and a quantitative phase involving distribution of a questionnaire to research participants and researchers. The data from the qualitative phase were used to assist with the development of the quantitative instrument. Descriptive statistics were derived to describe the various attitudes and practices with respect to the conduct of research. RESULTS: The qualitative study identified issues concerning monitoring procedures and the quality of communication between researchers and study participants. The quantitative analysis showed that parts of the Explanatory Statement (also known as the Participant Information Statement) were incomprehensible to 21% of research participants; the Explanatory Statement was considered too long by 34% of researchers; 6% of researchers believed that explicit consent was not always necessary; of the participants who were out of pocket for attending a study, 53% were offered compensation; and 44% of research participants were unaware of the existence of the ethics committee. In addition, 12% of researchers felt that the quality of monitoring should be improved. CONCLUSIONS: Improvements are necessary in the communication between ethics committees and researchers and research participants, and there is a need for more effective monitoring by ethics committees of research practices. The questionnaire designed for this study could be applied in a prospective manner as a useful tool for monitoring the conduct of research. PMID- 16472263 TI - How have we been managing chronic obstructive pulmonary disease in Australia? AB - AIM: Although chronic obstructive pulmonary disease (COPD) is a main cause of disability, hospital admissions and premature deaths in Australia, little is known about the community management of COPD in relation to recently published guidelines. The aim of the article was to report on COPD management in a community based cohort. METHODS: A random sample of adults aged between 45 and 70 years drawn from the electoral roll participated in the study. They completed a detailed respiratory questionnaire, spirometry, methacholine challenge and measurement of transfer factor. COPD was defined according to the Global initiative for Chronic Obstructive Lung Disease (GOLD) criteria. Current asthma was defined as wheeze during the last 12 months together with bronchial hyperreactivity. Subjects were classified as either COPD-only, asthma-only or both asthma and COPD. RESULTS: Of 1224 subjects completing spirometry, 39 (3.5%) met the GOLD criteria for stage 2 or 3 COPD, asthma-only was found in 99 (8.9%) subjects and 40 (3.6%) subjects had both asthma and COPD. The COPD-only group was significantly older than the other two groups. More than 40% of subjects with COPD did not have a diagnosis of COPD from their doctors. Only 48.7% of subjects with COPD had ever been prescribed medication for their breathing. More than two thirds of all subjects had seen a doctor for breathing problems, but very few had seen a general practitioner in the last 12 months and even fewer had respiratory function tests. CONCLUSIONS: Most subjects with COPD are being undertreated. Diagnosis, monitoring and referral systems should be improved. Preventive activities such as influenza vaccination and smoking cessation should be intensified. PMID- 16472264 TI - Management of chronic hepatitis B virus infection: a new era of disease control. AB - When assessing patients with chronic hepatitis B virus (HBV) infection, consider the state of viral replication, the immune response and whether viral mutations could be present, as well as evidence for liver disease or extrahepatic manifestations. In wild-type infections, loss of hepatitis B e antigen (HBeAg), gain of anti-HBe and disappearance of HBV DNA from serum indicate immunosuppression of viral replication, or 'nonreplicative chronic HBV infection'. This 'healthy carrier' state must be distinguished from HBeAg negative chronic hepatitis B (CHB) resulting from precore and core promoter mutations. HBeAg-negative CHB is common with genotypes D (Mediterranean region, south Asia) and C (north Asia) infections. Age, disease activity (alanine aminotransferase level) and severity (fibrosis stage, cirrhosis) influence treatment decisions. Following the marginal effectiveness of interferon and often temporary effectiveness of lamivudine due to drug resistance, treatment of CHB is entering a new era. Adefovir, entecavir, tenofovir, telbivudine and clevudine have equal or superior antiviral efficacy to lamivudine, whereas several agents are effective against lamivudine-resistant HBV. Pegylated-interferon (peginterferon) is superior to conventional interferon for obtaining sustained immunosuppression of HBV without drug resistance. Antiviral suppression of HBV replication for 2-5 years reverses hepatic fibrosis, prevents cirrhosis and, when cirrhosis is established, improves liver function, prevents hepatic decompensation and lowers the risk of liver cancer. Before embarking on immunosuppressive chemotherapy or organ transplantation in patients with chronic HBV infection, it is important to start antiviral therapy to prevent hepatitis flares. Antiviral therapy can be effective against membranous glomerulonephritis and polyarteritis nodosa caused by HBV. Further improvements in treatment of CHB are needed to prevent drug resistance and permanently suppress viral replication by eradicating viral templates or stimulating host immune responsiveness to HBV. PMID- 16472265 TI - Cardiac resynchronisation therapy for heart failure. AB - Heart failure (HF) is increasingly common and, despite advances in pharmacotherapeutic management, often progresses. Progression is marked by structural and electrical changes-remodelling. In approximately one-third of patients, ventricular dilatation is accompanied by intraventricular conduction delays, most commonly the left bundle branch block (LBBB). The presence of LBBB is associated with mechanical dyssynchrony of the heart. Cardiac resynchronisation therapy (CRT), the use of special pacemakers with or without implantable cardioverter defibrillators, aims to resynchronise the failing heart, improving myocardial contraction without increased energetics. Several, large, randomised clinical trials have now established the benefit of CRT in a select group of HF patients, providing functional and, recently shown, mortality benefits. However, a substantial proportion of patients are considered non responders to CRT, and studies are now underway to identify the patients most likely to respond to CRT. PMID- 16472266 TI - Consent in crisis: the need to reconceptualize consent to tissue banking research. AB - The issues surrounding consent to tissue banking research in Australia are complex and have created a forum of intense debate, thus providing a window of opportunity to critically appraise and challenge standard models of consent for research in general and for tissue banking research in particular. The usual practical difficulties associated with meeting the criteria for valid consent to research (including adequate information provision and voluntariness) are amplified in the case of tissue banking research. A number of models, based on widely accepted ethical principles, have been proposed to improve the process of obtaining consent to tissue banking research, all of which assume that the consent of individual tissue donors is needed to meet the criteria for valid consent. Feminist and communitarian theories use many of the same criteria for valid consent but interpret these criteria differently and de-emphasize the importance of individual autonomy as the central criterion for valid consent. An enriched model of consent incorporating feminist and communitarian ideas could satisfy the currently accepted criteria for valid consent while also furthering a broader range of community values. PMID- 16472267 TI - An apparently sporadic paraganglioma with an SDHB gene germline mutation presenting at age 68 years. AB - Paragangliomas (PGLs) are rare tumours arising from parasympathetic-associated paraganglia (particularly of the head and neck) or from sympathetic-associated paraganglia such as in the adrenal medulla when they are termed phaeochromocytomas and at extra-adrenal sites in the abdomen and thorax. Recent reports have found frequent germline mutations of VHL, RET, SDHB or SDHD not only in familial cases but also in apparently sporadic cases of phaeochromocytoma. These germline mutations are particularly likely to be found if multifocal disease is present or if the phaeochromocytoma or PGL occurs at a young age. We report a germline splice site mutation in SDHB in a patient presenting with an incidental, apparently sporadic, abdominal sympathetic PGL at 68 years of age. PMID- 16472268 TI - Medication accuracy and general practitioner referral letters. AB - Prescriber medication communication is a Quality Use of Medicines barrier. Medication information in General Practice (GP) referral letters to a physician was evaluated. Accuracy of medications taken and drug dose was respectively 63% and 84%, an overall accuracy rate of 58%. Complementary/over-the-counter medication documentation occurred in 26% of the letters. To avoid medical mismanagement, physicians must validate all GP medication lists regardless of their apparent comprehensiveness. PMID- 16472270 TI - Obstructive sleep apnoea induced atrial fibrillation. PMID- 16472269 TI - Clinical findings in Niemann-Pick disease type B. PMID- 16472271 TI - Cardiac amyloidosis--sustained clinical and free light chain response to low dose thalidomide and corticosteroids. PMID- 16472272 TI - Stool antigen testing for the diagnosis and confirmation of eradication of Helicobacter pylori infection. PMID- 16472275 TI - Drug-induced QT prolongation: an update. PMID- 16472276 TI - Chronotropic incompetence and abnormal autonomic modulation in ambulatory Chagas disease patients. AB - BACKGROUND: Chagas disease (ChD) patients might present chronotropic incompetence during exercise, although its physiopathology remains uncertain. We evaluated the heart rate (HR) response to exercise testing in ChD patients in order to determine the role of autonomic modulation and left ventricular dysfunction in the physiopathology of chronotropic incompetence. METHODS: ChD ambulatory patients (n = 170) and healthy controls (n = 24) underwent a standardized protocol including Doppler echocardiography, Holter monitoring, HR variability analysis, brain natriuretic peptide (BNP) measurement, and maximal exercise testing. The chronotropic response was calculated as the percentage of predicted HR achieved and the HR increment (DeltaHR) during exercise. ChD patients were divided according to the absence or presence of cardiopathy and chronotropic incompetence (<85% predicted HR). RESULTS: Chronotropic incompetence was present in 34 (20%) of all ChD patients. The group with cardiopathy displayed reduced DeltaHR (91 +/- 19 bpm) during exercise in comparison with ChD patients without cardiopathy (100 +/- 19 bpm). Both the values observed in ChD groups were significantly different from those of controls (112 +/- 13 bpm). Exercise duration, maximal oxygen consumption, and systolic blood pressure increment were significantly reduced in patients with abnormal chronotropic response. DeltaHR during the exercise was significantly correlated with markers of autonomic control of sinus node, such as rest HR (r =-0.498, P or=35%. The number of patients who received ICDs in this expanded MADIT-I subset also grew from 5.6% in 1994 to 14.6% in 2000. Mortality during a mean follow-up of 4.7 years was significantly higher in the MADIT-I group than in the expanded MADIT-I, or the remaining primary prevention and secondary prevention subsets. CONCLUSION: The MADIT-I has not only led to an increase in the number of patients undergoing prophylactic ICD implantation, but in clinical practice it has also been extrapolated to a broader population that has a different degree of risk than originally studied. PMID- 16472277 TI - High-resolution analysis of the surface P wave as a measure of atrial electrophysiological substrate. AB - BACKGROUND: At present atrial electrophysiology can only be assessed by invasive study. This limits available data in humans concerning atrial electrophysiologic changes in disease and in response to intervention. Indirect evidence suggests that the signal-averaged P wave (SAPW) may provide noninvasive markers of atrial electrophysiology but no direct evaluations that measure both refractoriness and conduction time have been reported. METHODS: We investigated 9 patients attending for diagnostic electrophysiological studies (4 male; mean age 35.7 years). A 20 pole catheter was positioned in the right atrium; a decapole catheter was placed in the coronary sinus. Atrial effective refractory period (AERP) and conduction times were measured at the lateral and septal right atrium and the left atrium during sinus rhythm (SR) and at pacing cycle lengths of 600, 500, and 400 ms. Simultaneous SAPW recordings were taken during SR and pacing at 600 ms. Intravenous flecainide (2 mg/kg) was given after which the protocol was repeated. RESULTS: Flecainide slowed conduction significantly at all sites (P < 0.05). During baseline measurements, rate adaptation of AERP was observed (P < 0.02 at the septum). Flecainide increased filtered P wave duration (P < 0.05) and reduced P wave energies (P < 0.05). Negative correlation was observed between P wave energies and conduction time with an inverse relationship between high-frequency energy and left atrial AERP. CONCLUSIONS: The SAPW provides a noninvasive marker of atrial electrophysiology. PMID- 16472279 TI - Prognostic significance of electrocardiographic abnormalities in diphtheritic myocarditis after hospital discharge: a long-term follow-up study. AB - OBJECTIVE: We aimed to investigate the long-term cardiac mortality and the relationship between cardiac mortality and electrocardiographic abnormalities in patients with diphtheritic myocarditis who survived after hospital discharge. MATERIALS AND METHODS: Between 1991 and 1996, 32 patients (all males, mean age 21.00 +/- 3.77 years) surviving diphtheritic myocarditis were included in the study and they were followed up for an average of 16.3 months (range 10.3-26.8 months) after hospital discharge. Clinical evaluation, ECG, and echocardiography were performed on admission, daily while in hospital and at the time of discharge. ECG changes were permanent during the follow-up period. The causes of death of the patients during follow-up period were inferred from the death records of the patients and talking to the people witnessing cardiac arrest. RESULTS: We observed that the patients with left bundle branch block (LBBB) and T wave inversion at hospital discharge had lower survival rates than that of the patients without these ECG changes in the long term. Although univariate Cox regression analysis identified LBBB (P = 0.001) and T wave inversion (P = 0.014) as the predictors of survival, only LBBB was an independent predictor of survival in multivariate Cox regression analysis. Adjusted hazard ratio was calculated as 13.67 for LBBB (P = 0.001; CI = 2.81-66.28). CONCLUSION: Diphtheritic myocarditis does not only demonstrate a malignant clinical course during acute phase of the disease, but also during the long-term follow-up period, especially in patients with LBBB and T wave inversion. Besides, T wave inversion and LBBB can help us to predict survival rate of the patients in long term. Moreover, LBBB is an independent predictor of long-term survival in diphtheritic myocarditis. PMID- 16472280 TI - Effects of atropine and pirenzepine on heart rate turbulence. AB - BACKGROUND: It has been shown that mortality risk in patients after myocardial infarction could be estimated by heart rate turbulence (HRT), a short-term change in heart rate after ventricular premature beat (VPB), presumably caused by baroreceptor mechanism. We sought to determine whether pharmacological blockade with atropine, or augmentation of vagal tone with pirenzepine given in small doses would influence HRT. METHODS: In 30 patients with normal echocardiogram, and without signs or symptoms of coronary artery disease, after electrophysiologic examination or radiofrequency ablation for supraventricular arrhythmias was completed, turbulence onset (TO) and turbulence slope (TS) in basal state, after 1.3 mg IV pirenzepine and finally, after atropine in dose of 0.04 mg/kg of body weight were compared. RESULTS: As assessed by Friedman ANOVA test both pirenzepine and atropine caused a significant change in both TO (P < 0.01) and TS (P < 0.01). The mean basal TO of -3.6 +/- 2.9%, changed after pirenzepine to -5.99 +/- 5.6% (P < 0.01), and after atropine it changed to -3.3 +/- 18.1% (P < 0.01). The mean basal TS of 18.6 +/- 10.1 ms/R-R interval increased after pirenzepine to 26.8 +/- 19.9 ms/R-R interval (P < 0.05), and decreased after atropine to 1.2 +/- 0.8 ms/R-R interval (P < 0.01). Mean cycle length increased after pirenzepine from 706.8 +/- 106.8 to 830 +/- 151.9 ms (P < 0.01), and decreased after atropine to 454.2 +/- 58.1 ms (P < 0.01). CONCLUSION: A conclusion could be drawn that vagomymetic manipulation with intravenous pirenzepine increases HRT; vagal blockade with atropine decreases HRT. This finding suggests that a normal vagal innervation of heart is a prerequisite for the phenomenon of HRT. PMID- 16472281 TI - Heart rate variability in patients with systemic sarcoidosis. AB - PURPOSE: The identification of subjects with systemic sarcoidosis at higher risk for sudden death is an unresolved issue. An influence of the autonomic activity on the genesis of ventricular arrhythmias was postulated. Heart rate variability (HRV) analysis provides a useful method to measure autonomic activity, and is a predictor of increased risk of death in various conditions. Therefore, the aim of the study was to evaluate HRV in patients with systemic sarcoidosis. METHODS: The study included 35 patients with biopsy proven systemic sarcoidosis who were not taking antiarrhythmic medications. Thallium scintigraphy was performed to all patients with systemic sarcoidosis. The cardiac sarcoidosis was accepted in 16 patients as abnormal thallium scintigraphy and normal coronary arteriography. The time-domain analysis of HRV was expressed as the standard deviation of all normal to normal NN intervals (SDNN) detected during 24-hour Holter monitoring. Twenty four healthy subjects represented a control group for HRV analysis. RESULTS: There were no differences in age (44 +/- 13 years for cardiac sarcoidosis, 42 +/- 15 years for noncardiac sarcoidosis, and 40 +/- 10 years for control group; P = NS), sex (the ratio of female; 63%, 68%, and 55%, respectively; P = NS), and echocardiographic ejection fraction (63 +/- 10%, 67 +/- 8%, and 69 +/- 6%, respectively; P = NS) among study groups. The mean SDNN value of the group with cardiac sarcoidosis was significantly lower than both the group with noncardiac sarcoidosis and the control group (72 +/- 32 ms vs 110 +/- 46 ms and 152 +/- 36 ms; P < 0.05, respectively). CONCLUSION: HRV is decreased in patients with systemic sarcoidosis compared to the control group. This decreasing is more obvious in patients with cardiac sarcoidosis. PMID- 16472282 TI - The association of QT dispersion and QT dispersion ratio with extent and severity of coronary artery disease. AB - BACKGROUND: Although prolongation of the QT intervals in acute ischemic conditions, such as acute myocardial infarction, intracoronary balloon inflation, and exercise induced ischemia, has been shown, association of rest QT intervals with extent and severity of stable coronary artery disease (CAD) has not been assessed so far. The effects of extent and severity of stable CAD on rest QT interval were analyzed in this study. METHODS: Rest 12-lead electrocardiograms (ECG) were recorded in 162 clinically stable subjects undergoing coronary angiography before the angiography for measurement of corrected QT dispersion (cQTd) and the QT dispersion ratio (QTdR) defined as QT dispersion divided by cycle length and expressed as a percentage. Angiographic "vessel score,""diffuse score," and "Gensini score" were used to evaluate the extent and severity of coronary atherosclerosis. Subjects were grouped as follows: those with normal angiogram (Group 1), those with insignificant (<50%) coronary stenosis (Group 2), and those with 1- (Group 3), 2- (Group 4), or 3-vessel disease (Group 5). RESULTS: cQTd and QTdR were higher in Group 3 compared with Group 1 (P < 0.001 and P = 0.001, respectively), in Group 4 compared with Group 1 (P < 0.001 for both) and Group 2 (P = 0.001 and P = 0.003, respectively), and in Group 5 compared with Group 1 (P < 0.001 for both) and Group 2 (P < 0.001 and P = 0.003, respectively). cQTd and QTdR were positively correlated with the vessel score (r = 0.422, P < 0.001; r = 0.358, P < 0.001, respectively), diffuse score (r = 0.401, P < 0.001; r = 0.357, P < 0.001, respectively) and Gensini score (r = 0.378, P < 0.001; r = 0.373, P < 0.001, respectively). In multiple linear regression analyses, cQTd was found to be independently associated only with diffuse score (beta= 0.325, P = 0.038). Also, QTdR was independently associated with diffuse score (beta= 0.416, P = 0.006) and Gensini score (beta= 0.374, P = 0.011). CONCLUSIONS: Rest cQTd and QTdR are increased, and related to the extent and severity of coronary atherosclerosis in patients with stable CAD. PMID- 16472283 TI - Effects of three fluoroquinolones on QT analysis after standard treatment courses. AB - BACKGROUND: Fluoroquinolone (FQ) agents have been speculated to influence the risk of Torsades de pointes (Tdp). Methods of evaluating this risk are varied and not systematic. QTc interval (QTc) prolongation is the most commonly used marker of Tdp, but has questionable utility. QT dispersion (QTd) may be a more selective marker of Tdp. No assessment of QTd for FQs has been reported. The current study evaluates the effects of three commonly prescribed FQs by comprehensive QT analysis. METHODS: In an open-label crossover study, 13 healthy participants received 3 treatments in random order: ciprofloxacin 500 mg twice daily, levofloxacin 500 mg once daily, and moxifloxacin 400 mg once daily. Each treatment was given for 7 days with a 1-week washout period. Twelve-lead electrocardiographic measurements were performed prior to the first dose, 2 hours after the first dose, and following the 7-day medication course. QTc prolongation was determined by measurement of lead II, and QTd from the difference between the maximum and minimum QTc intervals among the 12 leads. The data were analyzed using Friedman ANOVA, with the Wilcoxon signed rank test post hoc analysis, with P < 0.05 significance. RESULTS AND CONCLUSIONS: No difference was seen in baseline QTc (P = 0.48) or QTd (P = 0.92). Following 7 days of moxifloxacin, the QTc was prolonged by 6 ms relative to baseline (408 ms, P = 0.022), and 11 ms from the 2-hour measurement (403 ms, P = 0.003). Ciprofloxacin and levofloxacin had no effect on QTc, and no FQ changed the QTd. Within our study population, ciprofloxacin and levofloxacin did not display an increased risk for Tdp. Moxifloxacin, while showing QTc prolongation, did not affect QTd, and an increased Tdp risk is questionable. PMID- 16472284 TI - Functional significance of KCNH2 (HERG) K897T polymorphism for cardiac repolarization assessed by analysis of T-wave morphology. AB - BACKGROUND: Although KCNH2 (HERG) K897T polymorphism has been shown to be associated with the QT interval measured from 12-lead electrocardiogram (ECG), the functional significance of K897T polymorphism has been debated. The aim of this study was to test whether the K897T polymorphism of the KCNH2 (HERG) gene coding for the rapidly activating delayed rectifier K+ channel influences cardiac repolarization assessed by principal component analysis (PCA) of T-wave morphology. METHODS: Twelve-lead ECGs were digitized and T-wave morphology was analyzed with a PCA method in a population consisting of 228 healthy middle-aged subjects (121 women and 107 men). DNA samples were genotyped for the nucleotide 2690 A>C variation of the KCNH2 gene, corresponding to the KCNH2 K(lysine)897T(threonine) amino acid polymorphism. RESULTS: The allele frequencies were 0.86 (K) and 0.14 (T). The KCNH2 K897T polymorphism was associated with the total cosine R-to-T (TCRT), which reflects the wave front direction between depolarization and repolarization. TCRT was 0.421 in the genotype KK and 0.300 in the genotypes KT and TT (P = 0.04). The difference of TCRT was more marked between the KCNH2 K897T genotypes in women (P = 0.03) than in men (P = 0.52). CONCLUSIONS: The common K897T polymorphism of the cardiac potassium channel KCNH2 has functional significance for cardiac electrical properties. Subjects with a less common genotype, KT or TT, have smaller TCRT, which reflects dyssynchrony between depolarization and repolarization and is associated with an increased risk of cardiac mortality. PMID- 16472285 TI - The 12-lead ECG in patients with Mahaim fibers. AB - The aim of this review article is to discuss the electrocardiographic presentation of the so called variants of pre-excitation ("Mahaim fibers") during sinus rhythm and tachycardia. PMID- 16472287 TI - The history, science, and innovation of Holter technology. AB - Holter technology has endured for more than 40 years, and proven to be a valuable adjunctive noninvasive diagnostic technology to record the ambulatory or long term electrocardiogram in the study of living creatures. During this span of time, many scientists, physicians, and innovators contributed to the development and evolution of Holter technology. This essay seeks to document a view of the history and evolution of the technology during that time, and concomitantly give recognition to the scientists, physicians, and engineers who contributed so greatly. PMID- 16472288 TI - Long R-P' paroxysmal supraventricular tachycardia: what is the mechanism? AB - Deciphering the electrophysiologic mechanism of a long R-P' paroxysmal supraventricular tachycardia typically requires an invasive electrophysiologic study. We present a case where analysis of a simple surface 12-lead ECG was sufficient for the diagnosis. PMID- 16472289 TI - Canine renal transplantation: the next steps. PMID- 16472290 TI - Results of clinical renal transplantation in 15 dogs using triple drug immunosuppressive therapy. AB - OBJECTIVE: To evaluate outcome of renal transplantation in dogs administered cyclosporine, azathioprine, and prednisolone immunosuppression. STUDY DESIGN: Prospective clinical study. ANIMALS: Fifteen dogs with chronic renal failure. RESULTS: Nine dogs died within 1 month of surgery; 5 died from complications associated with generalized thromboembolism. Three dogs survived for 6-25 months. Three dogs alive at the time of this report have survived 22-48 months; however, all 3 dogs have had bacterial infections that responded to antibiotic therapy. There was no biochemical evidence of acute allograft rejection in any dog. Perioperative use of enoxaparin may have prevented thromboembolism in 5 dogs. CONCLUSIONS: Triple drug immunosuppressive therapy used in this study prevented acute renal allograft rejection in 6 dogs that survived >4 weeks; however, immunosuppression was excessive, resulting in an unacceptable frequency of infection and other drug-related complications. Perioperative anticoagulation therapy seem to be warranted. CLINICAL RELEVANCE: Survival time and quality of life for this group of dogs was poor; however, there was no evidence of acute rejection in the dogs surviving >4 weeks. This protocol should only be used if the degree of immunosuppression is reduced, and early evidence of allograft rejection is monitored by renal biopsy or markers of lymphocyte activation. PMID- 16472291 TI - Use of capecitabine after renal allograft transplantation in dog erythrocyte antigen-matched dogs. AB - OBJECTIVES: To investigate the use of a capecitabine (CAP)-based regimen after renal transplantation in dogs. STUDY DESIGN: Prospective, pilot study. ANIMALS: Healthy, unrelated, dog erythrocyte antigen (DEA)-matched, adult beagles. METHOD: Standard heterotopic renal transplantation with native nephrectomy was performed in 7 dogs. Dogs received oral, twice daily, CAP (250 mg/m2), cyclosporine-A (CsA) (4 mg/kg), ketoconazole (5 mg/kg), and prednisolone (0.25 mg/kg). After 90 days the surviving dogs were euthanatized and complete necropsy was performed. RESULTS: Seven transplants were performed. All dogs survived surgery. Six dogs had acute neurotoxicity, which resulted in death or euthanasia of 2 dogs within 2 days of surgery. In the remaining dogs, toxicity resolved rapidly with cessation of drug administration. Thereafter, modification of the regimen minimized toxicity. The 5 remaining dogs survived to study end; 4 dogs had no evidence of graft rejection. Necropsy examination was mostly unremarkable in all dogs. There were no major changes in CBC or biochemical values, except for a significant increase in serum calcium. CONCLUSIONS: CAP appeared well tolerated in most dogs. Toxicity occurred but abated with modification of the drug regimen. Efficacy for postoperative immunosuppression cannot be determined by this study, although results are promising. CLINICAL RELEVANCE: CAP-CsA-prednisolone is an effective, oral immunosuppressive regimen for prevention of acute allograft rejection in DEA matched beagles. Further studies on dose, toxicity, and efficacy compared with current immunosuppressive regimens are needed before use in clinical practice. PMID- 16472292 TI - Renal allograft histopathology in dog leukocyte antigen mismatched dogs after renal transplantation. AB - OBJECTIVE: To evaluate allograft histopathology in dog leukocyte antigen (DLA) mismatched dogs undergoing renal transplantation, with transient immunosuppression. STUDY DESIGN: Prospective study. ANIMALS: Ten healthy adult mongrel dogs. METHODS: Reciprocal renal transplantation and bilateral nephrectomy were performed. Immune conditioning consisted of nonmyeloablative (200 cGy), total body irradiation (TBI), bone marrow transplantation (BMT; 7 dogs), cyclosporine (CSA; 15 mg/kg every 12 hours), mycophenolate mofetil (MMF; 10 mg/kg every 12 hours) and intermittent prednisone (1 mg/kg every 12-24 hours). Biopsies were collected at transplantation, during full immunosuppression (44-90 days), and once medications were reduced or discontinued (228-580 days). Biopsies were evaluated for interstitial, tubular, vascular, and glomerular lesions. Blood urea nitrogen, creatinine, serum CSA concentrations, and clinical score were determined at each biopsy. RESULTS: Seven dogs survived >200 days (mean, 380 days). Transient CSA toxicity was suspected in 6 dogs. Lymphocytic, plasmacytic interstitial inflammation, and tubulitis progressed when immunosuppressive medications were decreased. All 7 dogs had histologic lesions consistent with some degree of allograft rejection at study end. CONCLUSION: Nonmyeloablative TBI, BMT, and short-term immunosuppression with CSA, MMF, and prednisone allowed renal allograft function and dog survival for >200 days. It appears unlikely that total drug withdrawal will be possible in unrelated DLA-mismatched dogs using this protocol. CLINICAL RELEVANCE: Transient immunosuppression with MMF, CSA, and prednisone along with BMT and nonmyeloablative TBI may make kidney transplantation a clinical reality for treatment of kidney failure in dogs. Initiating both MMF and CSA at lower dosages may potentially eliminate early renal allograft injury. PMID- 16472293 TI - Making a rational choice between ovariectomy and ovariohysterectomy in the dog: a discussion of the benefits of either technique. AB - OBJECTIVE: To determine if ovariectomy (OVE) is a safe alternative to ovariohysterectomy (OVH) for canine gonadectomy. STUDY DESIGN: Literature review. METHODS: An on-line bibliographic search in MEDLINE and PubMed was performed in December 2004, covering the period 1969-2004. Relevant studies were compared and evaluated with regard to study design, surgical technique, and both short-term and long-term follow-up. CONCLUSIONS: OVH is technically more complicated, time consuming, and is probably associated with greater morbidity (larger incision, more intraoperative trauma, increased discomfort) compared with OVE. No significant differences between techniques were observed for incidence of long term urogenital problems, including endometritis/pyometra and urinary incontinence, making OVE the preferred method of gonadectomy in the healthy bitch. CLINICAL RELEVANCE: Canine OVE can replace OVH as the procedure of choice for routine neutering of healthy female dogs. PMID- 16472294 TI - Biomechanics of tibial plateau leveling of the canine cruciate-deficient stifle joint: a theoretical model. AB - OBJECTIVE: To evaluate the effect of tibial plateau leveling on the biomechanics of the canine stifle. STUDY DESIGN: Analysis of a 3-dimensional (3-D) anatomically accurate theoretical model of the canine stifle. METHODS: A 3-D, 3 segment mathematical model of the normal canine stifle was modified to simulate the effect of rotation of the tibial plateau during tibial plateau leveling osteotomy (TPLO). The model examined the normal stifle, the stifle with a tibial plateau angle (TPA) of 0 degrees, and the stifle with a TPA of 5 degrees. Analysis of the models at 10 consecutive equally spaced positions during the stance phase yielded data such as ligament forces and joint reaction forces at each position. RESULTS: Rotation of the tibial plateau to a TPA of 0 degrees almost eliminates forces in the cranial cruciate ligament (CCL) throughout the stance phase. Rotation to a TPA of 5 degrees did not, however, substantially decrease the load in the CCL. Both procedures increased the load in the caudal cruciate ligament (CaCL). CONCLUSIONS: Cranial tibial thrust (CTT) is converted into caudal tibial thrust when the TPA is 0 degrees ; however, rotating the plateau to a TPA of 5 degrees does not eliminate the CTT. CLINICAL RELEVANCE: The TPLO procedure performed as currently recommended (rotating the tibial plateau to a TPA of 5 degrees) may not eliminate the CTT, but only reduce it. Both TPLO procedures evaluated here were found to increase the load in the CaCL. PMID- 16472295 TI - Cortical bone graft and endoprosthesis in the distal radius of dogs: a biomechanical comparison of two different limb-sparing techniques. AB - OBJECTIVE: To compare the biomechanical properties of cortical bone and surgical steel endoprosthesis for limb-sparing surgery of the distal radius in dogs and evaluate the role of the ulna in providing stability to the reconstructed limb. STUDY DESIGN: Cadaveric biomechanical study. ANIMALS: Twelve pairs of normal canine thoracic limbs. METHODS: Paired limbs were divided into 4 groups: endoprosthesis and cortical bone graft, with and without preservation of the ulna. In each limb pair, the distal segment of the radius resected from the limb to be reconstructed with an endoprosthesis was used as the cortical bone graft in the contralateral limb. The ulna was resected en bloc with the radius and at the same level as the radial osteotomy in limbs where the ulna was not preserved. Limbs were tested in axial loading until failure. The load-deformation curve was used to acquire the biomechanical properties of each construct, which were compared using 2-way ANOVA. Failure modes were compared descriptively. RESULTS: Limbs reconstructed with the endoprosthesis had significantly greater yield load, energy at yield, and ultimate load compared with limbs reconstructed with a cortical bone graft. There were no significant differences in either energy to failure or stiffness between the 2 constructs. Preservation of the ulna did not significantly improve any of the biomechanical properties tested with either endoprosthesis or cortical bone graft constructs. The modes of failure in all 4 groups were variable and inconsistent. CONCLUSIONS: Limbs reconstructed with an endoprosthesis were biomechanically superior to limbs reconstructed with a cortical bone graft in axial loading to failure. Preservation of the ulna is not required to improve the stability in axial compression after limb-sparing surgery of the distal radius. CLINICAL RELEVANCE: The endoprosthesis may provide another option for limb-sparing surgery of the distal radius in dogs. It has potential advantages when compared with cortical bone grafts, including better biomechanical performance and resistance to implant failure in axial compression, immediate availability, and no requirement for bone banking facilities. The ulna can be resected en bloc with the radius without having a negative impact on construct stability. En bloc resection of the ulna and radius may decrease the risk of local tumor recurrence after limb-sparing surgery. PMID- 16472296 TI - Ex vivo comparison of one versus two distal screws in 8 mm model 11 interlocking nails used to stabilize canine distal femoral fractures. AB - OBJECTIVE: To compare the structural properties of an 8 mm model 11 interlocking nail (IN) with 2 proximal and 2 distal screws (2/2) to 2 proximal and 1 distal screws (2/1) in an unstable canine fracture model. STUDY DESIGN: Ex vivo biomechanical investigation. SAMPLE POPULATION: Eight pairs of adult canine femurs. METHODS: A simple transverse distal metaphyseal femoral fracture with a 1 cm gap was created. The unstable fracture in 1 femur was repaired with a nail with 2 distal and 2 proximal screws and the paired femur with a nail with 1 distal and 2 proximal screws. Cyclic mechanical testing in torsion was performed to assess fatigue life, peak torque, stiffness, and mode of failure. RESULTS: All 2/1 IN-femoral constructs, but only 2 of eight 2/2 constructs, failed before completion of 50,000 loading cycles. The 2/2 constructs had significantly greater peak torque to failure (P = .002) and longer fatigue life (P = .00003) compared with 2/1 constructs. There were no significant differences in stiffness between 2/2 and 2/1 constructs when the non-failed constructs were compared (P > .5). All constructs failed by screw deformation. CONCLUSIONS: An 8 mm model 11 IN used for fixation of unstable canine distal femoral fractures has a longer fatigue life and is stronger under torsional loads when 2 rather than 1 distal screws are placed. CLINICAL RELEVANCE: When repairing unstable canine distal femoral fractures with an IN system, 2 distal screws should be inserted to avoid catastrophic implant failure before bone healing is achieved. PMID- 16472297 TI - Tibial nerve somatosensory evoked potentials in dogs with degenerative lumbosacral stenosis. AB - OBJECTIVE: To determine somatosensory evoked potentials (SEPs) in dogs with degenerative lumbosacral stenosis (DLS) and in healthy dogs. STUDY DESIGN: Clinical and experimental study. ANIMALS: Dogs with DLS (n = 21) and 11 clinically normal dogs, age, and weight matched. METHODS: Under anesthesia, the tibial nerve was stimulated at the caudolateral aspect of the stifle, and lumbar SEP (LSEP) were recorded percutaneously from S1 to T13 at each interspinous space. Cortical SEP (CSEP) were recorded from the scalp. RESULTS: LSEP were identified as the N1-P1 (latency 3-6 ms) and N2-P2 (latency 7-13 ms) wave complexes in the recordings of dogs with DLS and control dogs. Latency of N1-P1 increased and that of N2-P2 decreased as the active recording electrode was moved cranially from S1 to T13. Compared with controls, latencies were significantly delayed in DLS dogs: .8 ms for N1-P1 and 1.7 ms for the N2-P2 complex. CSEP were not different between groups. CONCLUSIONS: Surface needle recording of tibial nerve SEP can be used to monitor somatosensory nerve function of pelvic limbs in dogs. In dogs with DLS, the latency of LSEP, but not of CSEP, is prolonged compared with normal dogs. CLINICAL RELEVANCE: In dogs with lumbosacral pain from DLS, the cauda equina compression is sufficient to affect LSEP at the lumbar level. PMID- 16472298 TI - Spinal epidural empyema in seven dogs. AB - OBJECTIVE: To characterize the clinical signs, diagnostic and surgical findings, and outcome in dogs with spinal epidural empyema (SEE). STUDY DESIGN: Retrospective study. ANIMALS: Seven dogs. METHODS: Dogs with SEE between 1992 and 2001 were identified from a computerized medical record system. Inclusion criteria were: neurologic examination, vertebral column radiographs, myelography, antimicrobial culture and susceptibility of material collected surgically from the vertebral canal, a definitive diagnosis of SEE confirmed by surgery, and microscopic examination of tissue from the vertebral canal. RESULTS: Common signs were lethargy, fever, anorexia, apparent spinal pain, and paraparesis/plegia. Common laboratory abnormalities were peripheral neutrophilia, and neutrophilic pleocytosis in cerebrospinal fluid (CSF). Three dogs had concurrent discospondylitis and 1 of these had vertebral luxation. On myelography, extradural spinal cord compression was focal (2 dogs), multifocal (3), or diffuse (2). Bacteria were isolated not from CSF but from blood, surgical site, pleural fluid, or urine in 6 dogs. Dogs were administered antibiotics and had surgical decompression by hemilaminectomy. Five dogs improved neurologically and had a good long-term outcome. Two dogs were euthanatized, 1 because of worsening of neurologic signs and pneumonia, and the other because of herniation of a cervical intervertebral disc 1 month postoperatively, unrelated to the SEE. CONCLUSION: Dogs with SEE may have a good outcome when treated by surgical decompression and antibiotic administration. CLINICAL RELEVANCE: SEE should be included in a list of possible causes for dogs with fever, apparent spinal pain, and myelopathy. PMID- 16472299 TI - Femoral neuropathy in a dog with iliopsoas muscle injury. AB - OBJECTIVE: To report femoral nerve dysfunction caused by focal iliopsoas muscle injury and treated by performing tenectomy of the muscle insertion. STUDY DESIGN: Case report. ANIMALS: A 4-year-old, castrated male, English Mastiff. RESULTS: Iliopsoas muscle injury caused femoral nerve deficits and severe pain. Focal injury was not detected by ultrasonography or computed tomography but was visible on magnetic resonance (MRI) images. Tenectomy of the insertion of the iliopsoas muscle relieved signs of pain. CONCLUSIONS: Femoral nerve dysfunction may occur with iliopsoas muscle injury. CLINICAL RELEVANCE: Iliopsoas muscle injury should be considered in patients with hip pain and MRI is a useful test for diagnosis. Tenectomy may be indicated for relief of chronic iliopsoas muscle pain. PMID- 16472300 TI - Combined spinal-epidural anesthesia in a dog. AB - OBJECTIVE: To report use of combined spinal epidural anesthesia for tail resection and surgical exploration of the pelvic canal and the perineal pararectal area in a dog. ANIMAL: A 4-month-old, 13 kg male Collie dog. METHODS: Under inhalant anesthesia, an epidural catheter was threaded through a Tuohy needle at L5-L6. Then using a Whitacre spinal needle bupivacaine and fentanyl were administered in the subarachnoid space at L6-L7 level. Fifteen minutes later, morphine was administered epidurally. Bupivacaine and morphine were administered epidurally 4 hours after the subarachnoid injection. RESULTS: No cardiorespiratory response to surgical stimulation was observed. Postoperative analgesia was satisfactory, and the catheter was removed 30 hours later. No complications or neurologic sequelae occurred before discharge or were noted 10 days later. CONCLUSION: Combined spinal-epidural anesthesia provided excellent intraoperative anesthesia and perioperative analgesia in a dog undergoing surgery involving the pelvic canal. CLINICAL RELEVANCE: Combined spinal-epidural anesthesia can be performed in dogs, and its use should be considered in major surgeries caudal to the diaphragm, as the epidural catheter allows cranial extension of the block, providing excellent intraoperative anesthesia and perioperative analgesia. PMID- 16472301 TI - Evaluation of laser Doppler flowmetry for measurement of capillary blood flow in the stomach wall of dogs during gastric dilatation-volvulus. AB - OBJECTIVES: To validate laser doppler flowmetry (LDF) for measurement of blood flow in the stomach wall of dogs with gastric dilatation-volvulus (GDV). ANIMALS: Six purpose-bred dogs and 24 dogs with naturally occurring GDV. STUDY DESIGN: Experimental and clinical. METHODS: Capillary blood flow in the body of the stomach and pyloric antrum was measured with LDF (tissue perfusion unit (TPU) before and after induction of portal hypertension (PH) and after PH plus gastric ischemia (GI; PH + GI) and compared with flow measured by colored microsphere technique. Capillary flow was measured by LDF in the stomach wall of dogs with GDV. RESULTS: PH and PH+GI induced a significant reduction in blood flow in the body of the stomach (P = .019). A significant positive correlation was present between percent changes in capillary blood flow measured by LDF and colored microspheres after induction of PH + GI in the body of the stomach (r = 0.94, P = .014) and in the pyloric antrum (r = 0.95, P = .049). Capillary blood flow measured in the body of the stomach of 6 dogs that required partial gastrectomy (5.00+/-3.30 TPU) was significantly lower than in dogs that did not (28.00+/ 14.40 TPU, P = .013). CONCLUSIONS: LDF can detect variations in blood flow in the stomach wall of dogs. CLINICAL RELEVANCE: LDF may have application for evaluation of stomach wall viability during surgery in dogs with GDV. PMID- 16472302 TI - Cesarean section and ovariohysterectomy after severe uterine torsion in four cows. AB - OBJECTIVE: To report a technique for intra-partum ovariohysterectomy (OVH) in cows after severe uterine compromise after uterine torsion. STUDY DESIGN: Clinical case report. ANIMALS: Four cows with uterine torsion. METHODS: After a ventrolateral cesarean section, uterine closure, and uterine detorsion, the laparotomy incision was elongated (20 cm) caudally so that the uterus can be completely exteriorized. The uterus was separated from broad ligaments by transection between 2 rows of overlapping interrupted sutures placed in the broad ligaments to a point approximately 2 cm caudal to the external os of the cervix. The vagina was occluded with 2 clamps directly caudal to the external os of the cervix and closed with 5 single cruciate sutures that penetrated all layers, and then the vagina was transected between the suture line and external os to remove the uterus. The abdominal incision was closed in layers. Results- All cows survived the surgery; however, 1 died the next day from Escherichia coli sepsis and 1 was euthanatized at 8 days because of unresponsive E. coli mastitis. One cow, 28 days after surgery developed a 15 cm perivaginal abscess, which resolved after drainage. This cow and the fourth cow survived and lactated. CONCLUSIONS: Intra partum OVH is technically possible after cesarean section in cows with uterine compromise or necrosis after uterine torsion. CLINICAL RELEVANCE: Ovariohysterectomy should be considered in cows with severe uterine torsion that have uterine tissue compromise to improve survival. PMID- 16472303 TI - Members of the IclR family of bacterial transcriptional regulators function as activators and/or repressors. AB - Members of the IclR family of regulators are proteins with around 250 residues. The IclR family is best defined by a profile covering the effector binding domain. This is supported by structural data and by a number of mutants showing that effector specificity lies within a pocket in the C-terminal domain. These regulators have a helix-turn-helix DNA binding motif in the N-terminal domain and bind target promoters as dimers or as a dimer of dimers. This family comprises regulators acting as repressors, activators and proteins with a dual role. Members of the IclR family control genes whose products are involved in the glyoxylate shunt in Enterobacteriaceae, multidrug resistance, degradation of aromatics, inactivation of quorum-sensing signals, determinants of plant pathogenicity and sporulation. No clear consensus exists on the architecture of DNA binding sites for IclR activators: the MhpR binding site is formed by a 15-bp palindrome, but the binding sites of PcaU and PobR are three perfect 10-bp sequence repetitions forming an inverted and a direct repeat. IclR-type positive regulators bind their promoter DNA in the absence of effector. The mechanism of repression differs among IclR-type regulators. In most of them the binding sites of RNA polymerase and the repressor overlap, so that the repressor occludes RNA polymerase binding. In other cases the repressor binding site is distal to the RNA polymerase, so that the repressor destabilizes the open complex. PMID- 16472304 TI - Genetic improvement of processes yielding microbial products. AB - Although microorganisms are extremely good in presenting us with an amazing array of valuable products, they usually produce them only in amounts that they need for their own benefit; thus, they tend not to overproduce their metabolites. In strain improvement programs, a strain producing a high titer is usually the desired goal. Genetics has had a long history of contributing to the production of microbial products. The tremendous increases in fermentation productivity and the resulting decreases in costs have come about mainly by mutagenesis and screening/selection for higher producing microbial strains and the application of recombinant DNA technology. PMID- 16472305 TI - Fungal laccases - occurrence and properties. AB - Laccases of fungi attract considerable attention due to their possible involvement in the transformation of a wide variety of phenolic compounds including the polymeric lignin and humic substances. So far, more than a 100 enzymes have been purified from fungal cultures and characterized in terms of their biochemical and catalytic properties. Most ligninolytic fungal species produce constitutively at least one laccase isoenzyme and laccases are also dominant among ligninolytic enzymes in the soil environment. The fact that they only require molecular oxygen for catalysis makes them suitable for biotechnological applications for the transformation or immobilization of xenobiotic compounds. PMID- 16472306 TI - Plasmids of lactococci - genetic accessories or genetic necessities? AB - Lactococci are one of the most exploited microorganisms used in the manufacture of food. These intensively used cultures are generally characterized by having a rich plasmid complement. It could be argued that it is the plasmid complement of commercially utilized cultures that gives them their technical superiority and individuality. Consequently, it is timely to reflect on the desirable characteristics encoded on lactococcal plasmids. It is argued that plasmids play a key role in the evolution of modern starter strains and are a lot more than just selfish replicosomes but more essential necessities of intensively used commercial starters. Moreover, the study of plasmid biology provides a genetic blueprint that has proved essential for the generation of molecular tools for the genetic improvement of Lactococcus lactis. PMID- 16472307 TI - Regulation of quorum sensing in Pseudomonas. AB - Bacteria use small signal molecules in order to monitor their population density and coordinate gene regulation in a process called quorum sensing. In Gram negative bacteria, the most common signal molecules are acylated homoserine lactones. Several Pseudomonas species produce acylated homoserine lactones that control important functions including pathogenicity and plant growth promotion. Many reports indicate that the quorum sensing systems of Pseudomonas are significantly regulated and interconnected with regulons of other global regulators. The integration of quorum sensing into additional regulatory circuits increases the range of environmental and metabolic signals beyond that of cell density, as well as further tuning the timing of the response. This review will focus on the regulation of quorum sensing in Pseudomonas, highlighting a complex response that might serve a given species to adapt in its particular environment. PMID- 16472308 TI - Protein secretion and secreted proteins in pathogenic Neisseriaceae. AB - Secreted proteins of pathogenic bacteria are often essential virulence factors. They are involved, for example, in the adherence of the bacteria to host cells or required to suppress the host's defence mechanisms. Until recently, only IgA1 protease had been studied in detail in the NeisseriaceaeNeisseria meningitidis and Neisseria gonorrhoeae. The availability of their genome sequences, however, has boosted research in this area. Here, we present a survey of the secretome of the pathogenic Neisseriaceae, based on the available genome sequences, and the current knowledge of the functions and structures of the secreted proteins. Of the six protein-secretion pathways that are widely disseminated among Gram negative bacteria, three pathways appear to be present among the Neisseriaceae, i.e. the autotransporter-, the two-partner- and the type I-secretion mechanisms. Comparison of the predicted secretomes reveals a considerable flexibility. As compared with N. meningitidis and the nonpathogen N. lactamica, N. gonorrhoeae appears to have a considerably degenerated secretome, which may reflect its altered niche occupancy. The flexibility of the secretome may be enhanced by the presence of ORFs in the genomes potentially encoding fragments of secreted proteins. We hypothesize that these ORFs may substitute for the corresponding fragments in the full-length genes through genetic recombination, thereby changing the host-cell receptor specificity of the secreted protein. PMID- 16472309 TI - Infants show a facilitation effect for native language phonetic perception between 6 and 12 months. AB - Patterns of developmental change in phonetic perception are critical to theory development. Many previous studies document a decline in nonnative phonetic perception between 6 and 12 months of age. However, much less experimental attention has been paid to developmental change in native-language phonetic perception over the same time period. We hypothesized that language experience in the first year facilitates native-language phonetic performance between 6 and 12 months of age. We tested 6-8- and 10-12-month-old infants in the United States and Japan to examine native and nonnative patterns of developmental change using the American English /r-l/ contrast. The goals of the experiment were to: (a) determine whether facilitation characterizes native-language phonetic change between 6 and 12 months of age, (b) examine the decline previously observed for nonnative contrasts and (c) test directional asymmetries for consonants. The results show a significant increase in performance for the native-language contrast in the first year, a decline in nonnative perception over the same time period, and indicate directional asymmetries that are constant across age and culture. We argue that neural commitment to native-language phonetic properties explains the pattern of developmental change in the first year. PMID- 16472310 TI - Birthweight-discordance and differences in early parenting relate to monozygotic twin differences in behaviour problems and academic achievement at age 7. AB - This longitudinal monozygotic (MZ) twin differences study explored associations between birthweight and early family environment and teacher-rated behaviour problems and academic achievement at age 7. MZ differences in anxiety, hyperactivity, conduct problems, peer problems and academic achievement correlated significantly with MZ differences in birthweight and early family environment, showing effect sizes of up to 2%. As predicted by earlier research, associations increased at the extremes of discordance, even in a longitudinal, cross-rater design, with effect sizes reaching as high as 12%. As with previous research some of these non-shared environmental (NSE) relationships appeared to operate partly as a function of SES, family chaos and maternal depression. Higher risk families generally showed stronger negative associations. PMID- 16472311 TI - Gaze following: why (not) learn it? AB - We propose a computational model of the emergence of gaze following skills in infant-caregiver interactions. The model is based on the idea that infants learn that monitoring their caregiver's direction of gaze allows them to predict the locations of interesting objects or events in their environment (Moore & Corkum, 1994). Elaborating on this theory, we demonstrate that a specific Basic Set of structures and mechanisms is sufficient for gaze following to emerge. This Basic Set includes the infant's perceptual skills and preferences, habituation and reward-driven learning, and a structured social environment featuring a caregiver who tends to look at things the infant will find interesting. We review evidence that all elements of the Basic Set are established well before the relevant gaze following skills emerge. We evaluate the model in a series of simulations and show that it can account for typical development. We also demonstrate that plausible alterations of model parameters, motivated by findings on two different developmental disorders - autism and Williams syndrome - produce delays or deficits in the emergence of gaze following. The model makes a number of testable predictions. In addition, it opens a new perspective for theorizing about cross species differences in gaze following. PMID- 16472312 TI - Blind infants in random environments: further predictions. PMID- 16472313 TI - Modeling the development of gaze following needs attention to space. PMID- 16472314 TI - The benefits of computational modelling for the study of developmental disorders: extending the Triesch et al. model to ADHD. PMID- 16472316 TI - Learning words: children disregard some pragmatic information that conflicts with mutual exclusivity. AB - Children tend to infer that when a speaker uses a new label, the label refers to an unlabeled object rather than one they already know the label for. Does this inference reflect a default assumption that words are mutually exclusive? Or does it instead reflect the result of a pragmatic reasoning process about what the speaker intended? In two studies, we distinguish between these possibilities. Preschoolers watched as a speaker pointed toward (Study 1) or looked at (Study 2) a familiar object while requesting the referent for a new word (e.g. 'Can you give me the blicket?'). In both studies, despite the speaker's unambiguous behavioral cue indicating an intent to refer to a familiar object, children inferred that the novel label referred to an unfamiliar object. These results suggest that children expect words to be mutually exclusive even when a speaker provides some kinds of pragmatic evidence to the contrary. PMID- 16472317 TI - Syllogistic reasoning and belief-bias inhibition in school children: evidence from a negative priming paradigm. AB - Research on deductive reasoning in adolescents and adults has shown that errors in deductive logic are not necessarily due to a lack of logical ability but can stem from an executive failure to inhibit biases. Few studies have examined this dissociation in children. Here, we used a negative priming paradigm with 64 children (8-10 years old) to test the role of cognitive inhibition in syllogisms with belief-bias effects. On trials where negative priming was predicted, results were as follows: For the first syllogism (A), the strategy 'unbelievable-equals invalid' had to be inhibited. The logic of the syllogism led to affirming a conclusion inconsistent with one's knowledge of the world, such as 'All elephants are light.' For the second syllogism (B), one's real-world knowledge and the syllogism's logic were congruent but the latter required affirming exactly what had been inhibited for A (i.e. that elephants are heavy). A negative priming effect on the A-B sequence was reflected in a significant drop in reasoning performance on B. This supports the idea that during cognitive development, inhibitory control is required for success on syllogisms where beliefs and logic interfere. PMID- 16472318 TI - Infants' physical knowledge affects their change detection. AB - Prior research suggests that infants attend to a variable in an event category when they have identified it as relevant for predicting outcomes in the category, and that the age at which infants identify a variable depends largely on the age at which they are exposed to appropriate observations. Thus, depending on age of exposure, infants may identify the same variable at different ages in different event categories. A good case in point is the variable height, which is identified at about 3.5 months in occlusion events, but only at about 12 months in covering events and 14 months in tube events. In the present experiments, 11 month-olds detected a change to an object's height in an occlusion but not a covering event, and 12.5-month-olds detected a similar change in a covering but not a tube event. Thus, infants succeeded in detecting a change to an object's height in an event where height had been identified as a relevant variable, but failed to detect the exact same change in another event where height had not yet been identified as a relevant variable. These findings provide evidence that infants' physical knowledge affects which changes they detect in physical events. Possible mechanisms underlying these findings are also discussed, in light of recent accounts of change detection in adults. PMID- 16472319 TI - Change in children's understanding of others' intentional actions. AB - The study investigated children's intention understanding using a longitudinal design. Thirty-two Italian children were tested on the 'Demonstration of Intention' in the Re-enactment paradigm devised by Meltzoff (1995a), at two ages. Mean age was 12 months at the first session and 15 months at the second session. Previous research by Meltzoff showed that 18-month-olds are able to re-enact what the adult intended to do, when an adult tried but failed to perform certain target acts. Bellagamba and Tomasello (1999) found that, compared to 18-month olds, 12-month-olds are less able to imitate unsuccessful goal-directed actions. The current study had two main goals. The first was to replicate earlier findings on intention understanding in 12- and 15-month-olds longitudinally assessed. The second goal was to analyse which actions children perform when not producing the target act. A coding scheme was devised to evaluate these non-target actions. Results confirmed previous findings, i.e. children's tendency to produce the target act was higher at an older age. At a younger age the most frequent response was 'undifferentiated manipulation' of target objects, followed by 'approximation to the target action'. Since children differed in their intention understanding performance at the first session, we decided to check whether these individual differences remained stable at the second session. Individual children with high intention understanding performance at 12 months tended to remain high at 15 months of age. PMID- 16472320 TI - The eyes have it: visual pop-out in infants and adults. AB - Visual search studies with adults have shown that stimuli that contain a unique perceptual feature pop out from dissimilar distractors and are unaffected by the number of distractors. Studies with very young infants have suggested that they too might exhibit pop-out. However, infant studies have used paradigms in which pop-out is measured in seconds or minutes, whereas in adults pop-out occurs in milliseconds. In addition, with the previous infant paradigms the effects from higher cognitive processes such as memory cannot be separated from pop-out and selective attention. Consequently, whether infants exhibit the phenomenon of pop out and have selective attention mechanisms as found in adults is not clear. This study was an initial attempt to design a paradigm that would provide a comparable measure between infants and adults, thereby allowing a more accurate determination of the developmental course of pop-out and selective attention mechanisms. To this end, we measured 3-month-olds' and adults' saccade latencies to visual arrays that contained either a + among Ls (target-present) or all Ls (target-absent) with set sizes of 1, 3, 5 or 8 items. In Experiment 1, infants' saccade latencies remained unchanged in the target-present conditions as set size increased, whereas their saccade latencies increased linearly in the target absent conditions as set size increased. In Experiment 2, adults' saccade latencies in the target-present and target-absent conditions showed the same pattern as the infants. The only difference between the infants and adults was that the infants' saccade latencies were slower in every condition. These results indicate that infants do exhibit pop-out on a millisecond scale, that it is unaffected by the number of distractors, and likely have similar functioning selective attention mechanisms. Moreover, the results indicate that eye movement latencies are a more comparable and accurate measure for assessing the phenomenon of pop-out and underlying attentional mechanisms in infants. PMID- 16472321 TI - The development of emotional face processing during childhood. AB - Our facial expressions give others the opportunity to access our feelings, and constitute an important nonverbal tool for communication. Many recent studies have investigated emotional perception in adults, and our knowledge of neural processes involved in emotions is increasingly precise. Young children also use faces to express their internal states and perceive emotions in others, but little is known about the neurodevelopment of expression recognition. The goal of the current study was to determine the normal development of facial emotion perception. We recorded ERPs in 82 children 4 to 15 years of age during an implicit processing task with emotional faces. Task and stimuli were the same as those used and validated in an adult study; we focused on the components that showed sensitivity to emotions in adults (P1, N170 and frontal slow wave). An effect of the emotion expressed by faces was seen on the P1 in the youngest children. With increasing age this effect disappeared while an emotional sensitivity emerged on N170. Early emotional processing in young children differed from that observed in the adolescents, who approached adults. In contrast, the later frontal slow wave, although showing typical age effects, was more positive for neutral and happy faces across age groups. Thus, despite the precocious utilization of facial emotions, the neural processing involved in the perception of emotional faces develops in a staggered fashion throughout childhood, with the adult pattern appearing only late in adolescence. PMID- 16472322 TI - Words and maps: developmental changes in mental models of spatial information acquired from descriptions and depictions. AB - People acquire spatial information from many sources, including maps, verbal descriptions, and navigating in the environment. The different sources present spatial information in different ways. For example, maps can show many spatial relations simultaneously, but in a description, each spatial relation must be presented sequentially. The present research investigated how these source differences influence the mental models that children and adults form of the presented information. In Experiment 1, 8-year-olds, 10-year-olds and adults learned the layout of a six-room space either from verbal descriptions or from a map. They then constructed the configuration and pointed to target locations. Participants who learned from the map performed significantly better than those who learned from the description. Ten-year-olds performed nearly as well as adults did. The 8-year-olds' mental models differed substantially from the older children's and adults' mental models. The younger children retained the sequential information but did not integrate the relations into a survey-like cognitive map. Experiment 2 demonstrated that viewing the shape of the configuration, without seeing the map in full, could facilitate 8-year-olds' use of the verbal information and their ability to integrate the locations. The results demonstrate developmental differences in the mental representation of spatial information from descriptions. In addition, the results reveal that maps and other graphic representations can facilitate children's spatial thinking by helping them to transcend the sequential nature of language and direct experience. PMID- 16472323 TI - Our children too: a history of the Black Caucus of the Society for Research in Child Development, 1973-1997. AB - The Black Caucus of the Society for Research in Child Development(SRCD) was founded in 1973 to address concerns about the portrayal of Black children in scientific research, the lack of participation of ethnic minority members in the governance structure of SRCD, and the perceived need for a mutual support system for minority scholars aspiring to productive careers in the child development field. In this monograph, early members of the Caucus describe its history through the first 25 years, in 15 chapters distributed among sections on Caucus history, teaching and mentoring, publications and research-related issues, and supportive academic institutions. Among the topics explored are the formation and goals of the Caucus, its structure and membership, Caucus members' achieving stature and influence within SRCD, mentoring through the Toddler and Infant Experiences Study (TIES), response to the Atlanta child murders,the successful Pre-Conferences, SRCD Monographs and milestone developmental publications by Caucus members, the role of African American scholars in research on African American children (including the use of an ecological approach to study family processes), linkages between theory,research, and practice in Project Head Start, the contributions of the University of Michigan and Howard University, and looking to the future for students. Appendices trace Caucus chronological history and identify early sustaining members. This volume celebrates the accomplishments of the Caucus while also revisiting challenges that have arisen both internally and through membership in the SRCD parent organization. Key thematic issues include: cultural deficit versus cultural difference; linkages between poverty, race, and empowerment; advocacy versus objectivity in scientific research; and how the cultural or racial identity of the researcher informs scientific knowledge. The collaborations of Caucus members and others in SRCD modified the lens through which children of racially and ethnically diverse backgrounds are portrayed in the scientific literature. PMID- 16472327 TI - Sneddon's syndrome: another migraine-stroke association? AB - Sneddon's syndrome refers to the enigmatic association of ischaemic stroke and livedo reticularis. We review the Sneddon's syndrome literature examining the association of this condition with headache, including migraine. Case reports and series are stratified into two groups based on headache reference. In the group without a reference to headache, there are 208 persons, with a female to male ratio of 3 : 1. In the headache reference group, there are 175 persons, with a female to male ratio of 3.5 : 1. The proportion with headache in this second group is 58% (102 individuals), with headache described as migraine in 28 (27.5%) of the headache subjects, including six with migraine with aura. The frequency of headache is not significantly higher in persons with positive anti-phospholipid antibodies compared with the negative cohort (43% vs. 32%, P = 0.07). A review of the histopathological, radiological and serological data in Sneddon's syndrome and migraine underscores the plausibility of an association. Considered in the context of increased risk of stroke with migraine, a higher frequency of livedo in migraineurs with stroke, and the association of migraine and livedo reticularis, the question of whether livedo reticularis may be a risk marker for stroke in migraineurs is an area for further study. PMID- 16472328 TI - Neck-tongue syndrome and related (?) conditions. AB - A search has been made for the neck-tongue syndrome. It started out with a systematic inquiry during the Vaga study of headache epidemiology during the years 1995-97. Two cases were detected relatively early during the study. This led to a scrutiny of such cases also in our hospital headache practice. The origin - and the basis of the study, nevertheless, was the Vaga study. In the Vaga study, where 1838 18-65-year-old parishioners were examined, there were four neck-tongue syndrome cases (N-TS), i.e. a prevalence of approximately 0.22%. N-TS may be more frequent than hitherto surmised. A variant was observed in one case; a young male: instead of numbness, a 'spasm' seemed to occur in the tongue. None of the four had at any time consulted their physician for their complaints. In our regular headache practice, two new cases were detected. In one of them and in one of the Vaga cases, there was a combination with ipsilateral cervicogenic headache (CEH). In N-TS, there may be both ipsilateral headache and upper extremity sensory phenomena, a constellation reminiscent of CEH. The possible pathogenetic relationship between N-TS and CEH is therefore discussed in some detail. PMID- 16472329 TI - Evaluating the IHS criteria for cluster headache--a comparison between patients meeting all criteria and patients failing one criterion. AB - Cluster headache (CH) is diagnosed according to criteria of the International Headache Society (IHS), but, in clinical practice, these criteria seem too restrictive. As part of a nation-wide study, we identified a group of patients who met all criteria minus one (IHS-CH-1), and assessed in which way they differed from CH patients meeting all criteria (IHS-CH). We performed a nation wide questionnaire study for CH and CH-like syndromes, including questions based on the IHS criteria, and additional features such as restlessness during attacks, nocturnal onset of attacks, circadian rhythmicity of attacks and response to treatment. IHS-CH and IHS-CH-1 patients were compared. Of 1452 responders to two questionnaires, 1163 were IHS-CH and 289 were IHS-CH-1. The majority of the IHS CH-1 patients were classified as such because their attacks exceeded 3 h (64%, median attack duration: 5 h), or came in a frequency of less than 1 per 2 days (16%). Age at onset was similar between the groups. The male to female ratio was 3.7 : 1 in the IHS-CH group and around 1.6 : 1 in the IHS-CH-1 groups (P < 0.005). Patients with attacks exceeding 3 h less often reported a circadian rhythmicity (IHS-CH-1: 49%, IHS-CH: 64%), episodic periodicity (IHS-CH-1: 65%, IHS-CH: 78%), nocturnal attacks (IHS-CH-1: 67%, IHS-CH: 78%), smoking (IHS-CH-1: 90%, IHS-CH: 80%) and restlessness during attacks (IHS-CH-1: 64%, IHS-CH: 76%) than IHS-CH patients (P < 0.005). Photo- or phono-phobia (IHS-CH-1: 67%, IHS-CH: 54%) and nausea (IHS-CH-1: 38%, IHS-CH: 27%) were more frequently reported by patients who reported to have attacks exceeding 3 h (P < 0.005). Similar proportions reported effect of verapamil on their attacks (IHS-CH-1: 54%, IHS-CH 61%). We conclude that average attack duration exceeding 3 h was frequently the reason for not fulfilling IHS CH criteria. Symptoms often accompanying CH such as restlessness, nocturnal attacks and an episodic attack pattern were relatively frequently present in IHS-CH-1 patients with longer attacks. These patients may therefore be diagnosed with CH. Attack frequency may not be a useful criterion for the diagnosis of CH. The upper limit of 3 h should be increased in future diagnostic criteria. PMID- 16472330 TI - Preference for rizatriptan 10-mg wafer vs. eletriptan 40-mg tablet for acute treatment of migraine. AB - Preference is a composite, patient-oriented endpoint incorporating efficacy, tolerability, formulation, and convenience of medications. The objective of this study was to compare patient preference for rizatriptan 10-mg wafer vs. eletriptan 40-mg tablet for acute treatment of migraine. In this multicentre, open-label, two-period, crossover study, out-patients were randomly assigned to treat the first of two moderate to severe migraines with rizatriptan or eletriptan and the second with the alternate therapy. Patients completed diary assessments at baseline and up to 24 h after taking study medication. At the last visit, patients completed a psychometrically validated preference questionnaire. A total of 372 patients (mean age 38 years, 85% female) treated two migraine attacks, and 342 patients (92%) expressed a preference for treatment. Significantly more (P < or = 0.001) patients preferred rizatriptan 10-mg wafer [61.1%; 95% confidence interval (CI) 55.7, 66.3] to eletriptan 40-mg tablet (38.9%; 95% CI 33.7, 44.3). The most common reason given for preference of either treatment was speed of headache relief. At 2 h, 80% and 69% of patients reported that rizatriptan and eletriptan, respectively, was convenient or very convenient to take (mean convenience score 1.99 vs. 2.31, respectively; P < or = 0.001). Both triptans were well tolerated. In this head-to-head study designed to evaluate global patient preference, significantly more patients preferred the rizatriptan 10-mg wafer to the eletriptan 40-mg tablet for acute treatment of migraine. The single most important reason for preference was speed of relief, consistent with results from previous preference studies. PMID- 16472331 TI - Clinical-biochemical correlates of migraine attacks in rizatriptan responders and non-responders. AB - The present study was aimed at verifying the clinical characteristics of a typical attack in 20 migraine patients, 10 responders and 10 non-responders to rizatriptan, and at investigating any differences in the levels of neuropeptides of the trigeminovascular or parasympathetic systems [calcitonin gene-related peptide (CGRP), neurokinin A (NKA) and vasoactive intestinal peptide (VIP) measured by radioimmunoassay methods in external jugular blood] between responders and non-responders. In all responders to rizatriptan, pain was unilateral, severe, and pulsating, and in five of them at least one sign suggestive of parasympathetic system activation was recorded. Five patients who were non-responders to rizatriptan referred bilateral and non-pulsating pain, even though severe in most of them. CGRP and NKA levels measured before rizatriptan administration were significantly higher in responders than in non responders (P < 0.0001 and P < 0.002, respectively). In the five patients with autonomic signs among rizatriptan responders, detectable VIP levels were found at baseline. One hour after rizatriptan administration, a decrease in CGRP and NKA levels was evident in the external jugular venous blood of rizatriptan responders, and this corresponded to a significant pain relief and alleviation of accompanying symptoms. VIP levels were also significantly reduced at the same time in the five patients with autonomic signs. After rizatriptan administration, CGRP and NKA levels in non-responder patients showed less significant variations at all time points after rizatriptan administration compared with rizatriptan responders. The present study, although carried out on a limited number of patients, supports recent clinical evidence of increased trigeminal activation associated with a better triptan response in migraine patients accompanied by parasympathetic activation in a subgroup of patients with autonomic signs. In contrast, the poor response seems to be correlated with a lesser degree of trigeminal activation, lower variations of trigeminal neuropeptides after triptan administration, and no evidence of parasympathetic activation at baseline. PMID- 16472332 TI - Comparison of clinical symptoms and magnetic resonance angiographic (MRA) results in patients with trigeminal neuralgia and persistent idiopathic facial pain. Medium-term outcome after microvascular decompression of cases with positive MRA findings. AB - Neurovascular compression (NC) seems to have been confirmed as the major cause of classical trigeminal neuralgia (TN). In spite of the large number of surgically positive cases, however, there are still cases where no vascular compression of the trigeminal nerve can be found. To evaluate whether NC could be demonstrated preoperatively, high-resolution magnetic resonance angiography (MRA) was performed in 287 consecutive patients with TN and persistent idiopathic facial pain (PIFP) on a 0.5-T and a 1-T MR unit. Depending on the clinical symptoms, the TN cases were divided into typical TN and trigeminal neuralgia with non-neuralgic interparoxysmal pain (TNWIP) groups. Microvascular decompression (MVD) was performed in 103 of the MRA-positive cases. The patients were followed up postoperatively for from 1 to 10 years. The clinical symptoms were compared with the imaging results. The value of MRA was assessed on the basis of the clinical symptoms and surgical findings. The outcome of MVD was graded as excellent, good or poor. The clinical symptoms were compared with the type of vascular compression and the outcome of MVD. The MRA image was positive in 161 (56%) of the 287 cases. There were significant differences between the clinical groups: 66.5% of the typical TN group, 47.5% of the TNWIP group and 3.4% of the PIFP group were positive. The quality of the MR unit significantly determined the ratio of positive/negative MRA results. The surgical findings corresponded with the MRA images. Six patients from the MRA-negative group were operated on for selective rhizotomy and no NC was found. Venous compression of the trigeminal nerve was observed in a significantly higher proportion in the background of TNWIP than in that of typical TN on MRA imaging (24.1% and 0.8%, respectively) and also during MVD (31.2% and 1.2%, respectively). Four years following the MVD, 69% of the patients gave an excellent, 23% a good and 8% a poor result. The rate of some kind of recurrence of pain was 20% in the typical TN and 44% in TNWIP group. The rate of recurrence was 57% when pure venous compression was present. The only patient who was operated on from the PIFP group did not react to the MVD. The clinical symptoms and preoperative MRA performed by at least a 1-T MR unit furnish considerable information, which can play a role in the planning of the treatment of TN. PMID- 16472333 TI - Biochemical changes in endocannabinoid system are expressed in platelets of female but not male migraineurs. AB - The endogenous cannabinoid anandamide (AEA) plays important roles in modulating pain. Head pain is an almost universal human experience, yet primary headache disorders, such as migraine without aura (MoA) or episodic tension-type headache (ETTH), can represent a serious threat to well-being when frequent and disabling. We assessed the discriminating role of endocannabinoids among patients with ETTH or MoA, and control subjects. We measured the activity of AEA hydrolase and AEA transporter, and the level of cannabinoid receptors in peripheral platelets from MoA, ETTH and healthy controls. Sixty-nine headache patients and 36 controls were selected. Diagnosis of headache type was made according to the International Headache Society criteria. We observed significant sex differences concerning AEA membrane transporter and fatty acid amide hydrolase activity in all groups. An increase in the activity of AEA hydrolase and AEA transporter was found in female but not male migraineurs. Cannabinoid receptors were the same in all groups. Here we show that the endocannabinoid system in human platelets is altered in female but not male migraneurs. Our results suggest that in migraineur women an increased AEA degradation by platelets, and hence a reduced concentration of AEA in blood, might reduce the pain threshold and possibly explain the prevalence of migraine in women. The involvement of the endocannabinoid system in migraine is new and broadens our knowledge of this widespread and multifactorial disease. PMID- 16472334 TI - Effect of sympathetic and parasympathetic mediators on the release of calcitonin gene-related peptide and prostaglandin E from rat dura mater, in vitro. AB - Although not without controversy, an influence of the autonomic nervous system in headache is a matter for current debate. A possible contact site of autonomic and sensory nerves is the dura mater, where they form a dense network accompanying blood vessels. We investigated interactions between autonomic and nociceptive fibres by measuring release of calcitonin gene-related peptide (CGRP) and prostaglandin E2 (PGE2) from the dura mater, in vitro. The parasympathomimetic agent carbachol did not change basal release of CGRP or PGE2, whereas it diminished release induced by a mixture of inflammatory mediators. Norepinephrine did not change induced release of CGRP or PGE2, nor basal release of CGRP. However, basal release of PGE2 was enhanced by norepinephrine, and this enhancement was reduced by serotonin through 5-HT(1D) receptors. We conclude that sympathetic transmitters may control nociceptor sensitivity via increased basal PGE2 levels, a possible mechanism to facilitate headache generation. Parasympathetic transmitters may reduce enhanced nociceptor activity. PMID- 16472335 TI - A sleep study in cluster headache. AB - Cluster headache (CH) is a primary headache with a close relation to sleep. CH presents a circa-annual rhythmicity; attacks occur preferably during the night, in rapid eye movement (REM) sleep, and they are associated with autonomic and neuroendocrine modifications. The posterior hypothalamus is the key structure for the biological phenomenon of CH. Our aim is to describe a 55-year-old man presenting a typical episodic CH, in whom we performed a prolonged sleep study, consisting of a 9-week actigraphic recording and repeated polysomnography, with evaluation of both sleep macrostructure and microstructure. During the acute bout of the cluster we observed an irregular sleep-wake pattern and abnormalities of REM sleep. After the cluster phase these alterations remitted. We conclude that CH was associated, in this patient, with sleep dysregulation involving the biological clock and the arousal mechanisms, particularly in REM. All these abnormalities are consistent with posterior hypothalamic dysfunction. PMID- 16472336 TI - Cardiorespiratory regulation in migraine. Results in children and adolescents and review of the literature. AB - To investigate autonomic regulation in juvenile migraine we studied 70 children and adolescents with migraine during the headache-free period and 81 healthy controls by cardiorespiratory function tests. Heart rate variability was analysed with time and frequency domain indices during spontaneous breathing at rest and during metronomic breathing. Changes of heart rate and blood pressure were studied during tilt-table test, active standing, Valsalva manoeuvre and sustained handgrip. We found significant differences in metronomic breathing, tilt-table test and Valsalva manoeuvre. We interpret our findings and results reported in the literature as pointing to a restricted ability of the system to rest, which supports therapies intending to further this ability. In autonomic tests, hyperreactivity in juvenile migraineurs changes to hyporeactivity and passive coping in adults. This might be explained by disturbances of raphe nuclei and the periaqueductal grey. It corresponds to psychological findings in juvenile migraineurs reporting hypersensitivity and repressed aggression and claiming learned helplessness. PMID- 16472337 TI - The practice pattern of migraine management among neurologists in Taiwan. AB - To assess the prevalence of migraine and the attitudes and practice patterns of Taiwanese neurologists regarding migraine management, we mailed the survey questionnaire to all of the actively practicing board-certified neurologists from the membership list of the Taiwan Neurological Association. Of the targeted 531 neurologists, 123 (23.2%) participated in this study. Thirty of the participants (27.6%) reported having migraine. Most neurologists (88.5%) felt that headache was an important part of their practice. Many neurologists (65.0%) used neuroimaging to evaluate patients with severe headache and 44.7% used electroencephalography for headache evaluation. Many participants knew that combination analgesic was a common cause of medication-overuse headache, but did not know that ergotamine, acetaminophen and triptans were possible aetiologies of medication-overuse headache. Our study suggests that awareness of medication overuse headache and the indications of neuroimaging should be stressed in Taiwan, and headache guidelines should be modified according to local factors. PMID- 16472338 TI - Forward head posture and neck mobility in chronic tension-type headache: a blinded, controlled study. AB - Forward head posture (FHP) and neck mobility were objectively assessed in 25 patients with chronic tension-type headache (CTTH) and 25 healthy controls. Side view pictures were taken in a sitting position to measure the cranio-vertebral angle. A cervical goniometer was employed to measure the range of all cervical motions. Patients with CTTH showed a smaller cranio-vertebral angle (45.3 degrees +/- 7.6 degrees) than controls (54.1 degrees +/- 6.3 degrees), thus presenting a greater FHP (P < 0.001). Patients also had lesser neck mobility for all cervical movements, except for right lateral flexion (P < 0.01). There was a positive correlation between the cranio-vertebral angle and neck mobility. Within the CTTH group, a negative correlation was found between the cranio-vertebral angle and headache frequency, but neck mobility did not correlate with headache parameters. Further research is needed to define a potential role of FHP and restricted neck mobility in the origin or maintenance of TTH. PMID- 16472339 TI - The risks of sumatriptan administration in patients with unrecognized subarachnoid haemorrhage (SAH). AB - Administration of sumatriptan in subarachnoid haemorrhage (SAH) patients, misdiagnosed as migraine patients, may induce symptomatic cerebral vasospasm with potentially dangerous consequences. Over a 5-year period, we observed three patients with a 3-15-year history of migraine, who received sumatriptan for acute headache. Two patients received 6 mg sumatriptan subcutaneously on days 4 and 6, and one patient 3 x 100 mg sumatriptan orally on day 1 after an acute headache episode. In all three cases, an alleviation of headache intensity from severe to moderate was observed. When headache recurred and meningeal signs appeared, SAH was diagnosed by computed tomography in all three cases. No neurological deficits occurred during the further course of the disease. In both patients with a SAH caused by an aneurysm, transcranial Doppler sonography demonstrated vasospasm of the basal cerebral arteries. An antinociceptive effect of sumatriptan can be observed in SAH patients in good clinical condition, which suggests a specific craniovascular antinociceptive action. This may lead to misdiagnosis as migraine and delayed appropriate diagnosis and treatment. PMID- 16472340 TI - A novel ATP1A2 mutation in a family with FHM type II. AB - Familial hemiplegic migraine (FHM) is a rare subtype of migraine with aura with an autosomal dominant pattern of inheritance. Six FHM families underwent extensive clinical and genetic investigation. The authors identified a novel ATP1A2 mutation (E700K) in three patients from one family. In the patients, attacks were triggered by several factors including minor head trauma. In one subject a 3-day coma developed after a cerebral angiography. Overall, the phenotype of the patients closely resembles that of previously reported cases of FHM type II. The E700K variant might be regarded as the cause of the disease in this family, but this was not tested functionally. PMID- 16472341 TI - Cardiovascular autonomic function tests in cluster headache. AB - While facial autonomic signs are prominent during cluster headache (CH) attacks, cardiovascular autonomic changes have been described in few CH patients. Cardiovascular autonomic function tests (AFT) can be used to assess general autonomic function in CH patients in different stages of the disease. We aimed to assess whether general autonomic function is changed in CH patients during a cluster period. AFT was performed both during a cluster period, but outside an actual attack, and outside a cluster period in 18 patients. Heart rate variability was studied at rest, during deep breathing, after standing up and during a Valsalva manoeuvre. Blood pressure (BP) changes were recorded at rest, during standing up and during sustained handgrip. Measurements during and outside the cluster period were compared using the paired t-test. AFT measurements revealed no significant differences between the two measurements, except for diastolic BP in rest, which was higher during the cluster period [80.3 (SD 12.2) vs. 74.8 (SD 9.0), P = 0.04]. Autonomic dysfunction during a cluster period, but outside an attack, does not include systemic cardiovascular control. PMID- 16472342 TI - Implication of augmented vasogenic leakage in the mechanism of persistent aura in sporadic hemiplegic migraine. AB - The aim of this study is to report a possible implication of augmented vasogenic leakage in the mechanism of prolonged aura in sporadic hemiplegic migraine. A 35 year-old woman with sporadic hemiplegic migraine presented with headache followed by right arm weakness, right visual field defect, aphasia and confusion that persisted for 1 week. During the acute stage, focal hyperaemia was seen in the left cerebral hemisphere corresponding to persistent aura symptoms. Augmented vasogenic leakage was demonstrated on delayed enhanced fluid-attenuated inversion recovery image. Magnetic resonance angiography showed dilation of the left middle cerebral artery. During the convalescent stage, such abnormal findings were not seen. Based on these results, we speculate that augmented vasogenic leakage from the leptomeningeal vessels, probably associated with activation of the trigeminovascular system, may delay the recovery of hemiplegic migraine aura. PMID- 16472343 TI - Botulinum toxin A for the treatment of greater occipital neuralgia and trigeminal neuralgia: a case report with pathophysiological considerations. PMID- 16472344 TI - Hemicrania continua: side alternation and response to topiramate. PMID- 16472345 TI - Prosopagnosia as symptom of migraine with aura: a case report. PMID- 16472346 TI - Tension-type headache with aura. PMID- 16472347 TI - Verapamil-responsive hemicrania continua in a patient with episodic cluster headache. PMID- 16472348 TI - Nummular headache: first Italian case and therapeutic proposal. PMID- 16472350 TI - Ophthalmoplegic migraine. PMID- 16472353 TI - The Aum Cult leader Asahara's mental deviation and its social relations. AB - This paper is a further discussion of the author's previous reports on the crimes committed by the Asahara-Aum Cult. First, the motivations of Asahara-Aum crimes, in particular those of murders, and their abnormality beyond the realm of criminology are ascertained. Second, psychopathology of the leader, Asahara, was considered, and it was assumed that Asahara was affected with the syndrome of pseudologia phantastica which was confirmed by his strange attitude during his trials. Third, the mental status of Asahara after the first trial was investigated, and some evidence was found that his mental status was so severe as to be 'unfit to plead' that psychiatric evaluation and treatment would be necessary. Fourth, the reasons for making people, in particular scientific elites, fascinated with Asahara and the Aum Cult were scrutinized and some clues were described. Finally, the problem that Aum has survived under the name of Aleph, how heavy capital punishment for Leader Asahara is appropriate, and that reparations for victims of Asahara-Aum crimes are urgent from the standpoint of social psychiatry, are discussed. PMID- 16472354 TI - Application of the Revised Infant Temperament Questionnaire for evaluating temperament in the Japanese infant: creation of an abridged Japanese version. AB - The Japanese version of the Revised Infant Temperament Questionnaire (RITQ) was implemented to re-evaluate factor structure of the temperament scale with the objective of simplification. The questionnaire was conducted on 1099 mothers of Japanese infants visiting health centers for their children's health examinations at 6 or 7 months. As a result, a 57-item abridged version was constructed consisting of the seven subscales: 'fear of strangers and strange situations', 'insensitivity to food', 'regularity of rhythm', 'manageability', 'activity level', 'persistence of attention', and 'insensitivity to touch'. Although the items did not break down into the traditional nine temperament dimensions extracted by Carey et al., a certain commonality was seen in factor structure compared with the results of factor analysis using the RITQ in studies in Japan and abroad. PMID- 16472356 TI - Psychological impact of verbal abuse and violence by patients on nurses working in psychiatric departments. AB - The aim of this study was to assess the psychological impact of verbal abuse or violence by patients on nurses working in psychiatry departments and to identify factors related to their impact. Survey sheets were distributed to a total of 266 nurses working at two hospitals, and replies were obtained from 232 of them. Because three of them had less than 1 month of experience working in the psychiatry department and four of them failed to answer all the questions, valid replies were obtained from 225 nurses. Among the 225 whose replies were valid, 141 nurses who replied that they had experienced verbal abuse or violence that left an impression on them remained as the subjects of the final analysis. The Impact of Event Scale-Revised (IES-R) was used to evaluate psychological impact. Of the nurses who had been exposed to verbal abuse or violence that left an impression, 21% had scores that exceeded the IES-R cut-off point (24/25), and low satisfaction with family support, and neuroticism on the Eysenck Personality Questionnaire-Revised were shown to have contributed to the psychological impact. Nurses working in psychiatry departments were shown to experience a severe psychological impact when exposed to verbal abuse or violence. These results suggest the need for mental care approaches for nurses working in psychiatry departments. PMID- 16472355 TI - Quantification analysis of 123I-IPT single photon emission computed tomography uptake in the striatum of parkinsonism with the standard magnetic resonance template. AB - This study evaluated the striatal specific binding ratio (SBR), the anterior to posterior ratio of the striatum (APR) and its reproducibility by employing a template-based registration (TBR) method using the coregistered to the standard T1 magnetic resonance (MR) template (SMRT) as a replacement for the MR image of each patient. The 123I-IPT single photon emission computed tomography (SPECT) images of 30 patients with Idiopathic Parkinson's disease (IPD) and 11 normal controls were analyzed. The region of interest (ROI) was positioned manually in the same slice showing the highest striatal activity using the manual ROI method, while the ROI were positioned automatically in the mid striatal slice of the SPECT image coregistered to the SMRT. The SBR obtained using the TBR method showed a strong correlation with those using the manual method in all groups: normal controls (r = 0.851, P = 0.001), early IPD (r = 0.841, P < 0.001), and severe IPD (r = 0.702, P = 0.007). The APR obtained by the TBR correlated with those using the manual method in only the early IPD (r = 0.72, P = 0.001), while those obtained using the manual method showed no correlation in the three groups (P > 0.05). The reproducibility (rmsCV) of the TBR method was 7.2% (normal controls, 5.2%; mild IPD, 4.2%; severe IPD, 10.8%), while the reproducibility of the manual method was 31% (normal controls, 19.7%; mild IPD, 21.7%; severe IPD, 46.2%). This shows that the use of 123I-IPT SPECT for assessing IPD is affected by the method used to position the striatal ROI. This study showed that the TBR method using the SMRT is useful in diagnosing the IPD and assessing the disease severity with a high reproducibility, indicating a possibility of using the TBR method as a good replacement for the manual method. PMID- 16472357 TI - Characteristics of the sensory-motor, verbal and cognitive abilities of preschool boys with attention deficit/hyperactivity disorder combined type. AB - The purpose of this study is to clarify the characteristics of sensory-motor, verbal and cognitive abilities of preschool boys with attention deficit/hyperactivity disorder (ADHD) in order to provide information for their treatment and education at preschool age by teachers and professionals. For this purpose, 46 Japanese boys with ADHD-combined type (ADHD-C) whose ages ranged from 45 to 72 months were examined using the Japanese version of the Miller Assessment for Preschoolers (JMAP), and were compared with 46 Japanese boys matched for age and gender in the normative samples that served as the standardizations for the JMAP. The results showed that the ADHD-C group was significantly lower than the normative sample group both on the Total score and on each Index score (P < 0.01) with the exception of the Non-verbal Index. In particular, the number of boys with ADHD-C scoring below the 5th percentile on the Foundation Index (i.e. fundamental sensory-motor tests) was the highest among all index scores. The ADHD C group had significantly lower scores than the normative sample group in equilibrium, postural control, fine motor of hand and tongue, motor praxis, articulation, memory related to the comprehension of long sentences, and visual construction. Because fundamental sensory-motor abilities were notably lower in the ADHD-C group than in the normative sample group, it is suggested that preschool boys with ADHD-C should be examined and treated for sensory-motor disabilities. PMID- 16472358 TI - Factors associated with alcohol consumption, problem drinking, and related consequences among high school students in Taiwan. AB - This research evaluated the risk factors associated with alcohol consumption, problem drinking, and related consequences among high school students in Taiwan. A total of 779 10th grade students from four randomly selected high schools in eastern Taiwan were included in the survey. Survey data were self-reported by students and analyzed using logistic regression methods. The results showed that alcohol consumption and problem drinking was 2.22-2.71-fold greater in male than in female adolescents. Parents and peer groups were the determinants influencing alcohol consumption, and these influences could be enhanced particularly by the drinking behaviors of fathers and peer groups, and the relationship of students with their peer groups. The probability of developing adolescent problem drinking was fourfold greater in students whose fathers had habits of drinking. Ethnicity had a notable impact on the risk of problem drinking. The occurrence of problem drinking among indigenous adolescents was 2.98-fold higher than among Hans, indicating a diversity of biological factors, social norms, and expectation for alcohol drinking between these two ethnic groups. This study suggests that a policy for preventing alcohol abuse among high school students should start by addressing the two major associated problems: a need to earn peer group recognition, and the lack of family support among at-risk students. PMID- 16472359 TI - Prepulse inhibition of acoustic startle response in mild cognitive impairment and mild dementia of Alzheimer type. AB - Amnestic mild cognitive impairment (MCI) describes the condition of memory impaired individuals who otherwise function well and do not meet the clinical criteria for dementia. Such individuals are considered to represent a transitional stage between normal aging and dementia of Alzheimer type (DAT). Neurobiologic changes in amnestic MCI, and their significance for psychophysiologic function, are poorly understood. In this study, the authors compared acoustic prepulse inhibition (PPI) between subjects with amnestic MCI and mild DAT to characterize sensorimotor gating. The acoustic startle reflex, which the authors measured using an accelerometer and electromyogram, involves whole-body movement and eye blink in response to a sudden loud noise (115 dB). PPI is inhibition of this reflex by a softer noise (prepulse; 85 dB) preceding the startle stimulus by 30 ms. PPI was examined in 30 controls, 20 subjects with amnestic MCI, and 20 subjects with mild DAT. Neither amnestic MCI nor mild DAT affected startle movement amplitude. Subjects with amnestic MCI showed significantly enhanced PPI (gating facilitation), while subjects with mild DAT exhibited significantly less PPI than controls (gating deficit). This pattern of PPI changes suggests that neuropathologic changes in the limbic cortex, mainly the entorhinal cortex, at the earliest stage of DAT might be responsible for PPI abnormalities via disturbed regulation of the limbic cortico-striato-pallido pontine circuitry. Startle PPI changes could be used as a biologic marker for amnestic MCI and mild DAT. PMID- 16472360 TI - Relation between social functioning and neurocognitive test results using the Optional Thinking Test in schizophrenia. AB - The current study investigated the relationship between clinical evaluations of social functioning and neurocognitive test results, including various fluency tests for assessing divergent thinking, in patients with schizophrenia. The Optional Thinking Test (OTT) was used to measure the ability of individuals to conceive of alternatives. This test assesses alternative thinking, or the capacity to generate solutions to problems. The current study examined 36 schizophrenia patients and 25 normal subjects using the Mini-Mental State Examination (MMSE), the Rey Auditory Verbal Learning Test (RAVLT), the Letter Cancellation Test (LCT), the Letter and Category Fluency Tests, and the OTT for neurocognitive assessment, as well as the Positive and Negative Syndrome Scale (PANSS), the Global Assessment of Functioning (GAF), the Social Functioning Scale (SFS), and the Life Assessment Scale for the Mentally Ill - Interpersonal Relations (LASMI-I) for clinical measures. The schizophrenia patients had significantly poorer performances on the MMSE, RAVLT, LCT (time), fluency tests, and OTT than the controls. In the OTT, the proportions of classified strategies were indistinguishable between the schizophrenia patients and the controls. Alternative thinking, as measured by the OTT, was correlated with verbal fluency and attention but was not correlated with the social functioning scores (GAF, SFS, LASMI-I), whereas the Means-Ends Problem-Solving was correlated with the GAF in schizophrenia. Patients with schizophrenia could conceive of the same categories of alternatives as healthy people, but could not conceive as many alternatives. PMID- 16472361 TI - Prefrontal cognition in schizophrenia and bipolar illness in relation to Val66Met polymorphism of the brain-derived neurotrophic factor gene. AB - The measures of prefrontal cognition have been used as endophenotype in molecular genetic studies. Brain-derived neurotrophic factor (BDNF) has been implicated in cognitive functions and in the pathogenesis of major psychoses. This study investigates the relationship between Val66Met polymorphisms of the BDNF gene and prefrontal cognitive function in 129 patients with schizophrenia and 111 patients with bipolar mood disorder. Cognitive tests included the Wisconsin Card Sorting Test (WCST), with such domains as number of perseverative errors, non perseverative errors, completed corrected categories, conceptual level responses, and set to the first category, and the N-back test, where mean reaction time and percent of correct reactions were measured. Genotyping for Val66Met BDNF polymorphism was done by polymerase chain reaction method. In schizophrenia, no relationship between Val66Met polymorphism of the BDNF gene and the results of the WCST was observed. Patients with Val/Val genotype had a higher percentage of correct reactions in the N-back test than those with the remaining genotypes. Bipolar patients with Val/Val genotype obtained significantly better results on three of five domains of the WCST. No relationship between BDNF polymorphism and the results of the N-back test was found in this group. A limitation to the results could be variable psychopathological state and medication during cognitive testing and lack of Hardy-Weinberg equilibrium in schizophrenia group. Val66Met polymorphism of the BDNF gene may be associated with cognitive performance on the WCST in bipolar mood disorder but not in schizophrenia. An association of this polymorphism with performance on the N-back test in schizophrenia and not in bipolar illness may suggest that in schizophrenia, the BDNF system may be connected with early phases of information processing. PMID- 16472362 TI - Lifetime posttraumatic stress disorder in Turkish alcohol-dependent inpatients: relationship with depression, anxiety and erectile dysfunction. AB - The aim of the present study was to evaluate the prevalence of lifetime posttraumatic stress disorder (PTSD) in Turkish male alcohol-dependent inpatients, and to investigate the relationship of lifetime PTSD diagnosis with anxiety, depression, hopelessness, erectile dysfunction and psychosocial problems related with alcohol dependency. Eighty-two male inpatients who met DSM-IV criteria for alcohol dependence and 48 subjects without substance use disorder as a control group were included in the study. Subjects were applied the Hamilton Depression Rating Scale (HAM-D), the Hamilton Anxiety Rating Scale (HAM-A), the Michigan Alcoholism Screening Test (MAST), the Beck Hopelessness Scale (BHS) and the International Index of Erectile Function (IIEF). Rate of lifetime PTSD diagnosis was found to be 26.8% among alcohol-dependent inpatients. The mean age of patients with lifetime PTSD was lower than in patients without this diagnosis, while there were no significant differences between these two groups in terms of age of first alcohol use, lifetime major depression, current depression, presence and severity of erectile dysfunction. Mean scores of HAM-D, HAM-A, BHS and MAST in the group with lifetime PTSD were significantly higher than the group without this diagnosis. There was a positive relationship between lifetime PTSD diagnosis and depression, anxiety, hopelessness and severity of psychosocial problems related to alcohol dependency, while there was no relationship between lifetime PTSD comorbidity and erectile dysfunction in alcohol-dependent patients. PMID- 16472363 TI - Family history of alcoholism and cognitive recovery in subacute withdrawal. AB - A family history of alcoholism has been demonstrated to be an important factor affecting cognitive function. However, no studies have yet been conducted to compare cognitive recovery of family history-positive (FH+) and family history negative (FH-) alcoholics in the subacute withdrawal period. To tackle this problem, a neuropsychological test battery consisting of six computerized tests was administered to 19 FH+ and 20 FH- alcoholics at 2 and 7 weeks after abstinence. At 2 weeks after abstinence, overall performance of both FH+ and FH- groups was significantly poorer than that of healthy controls. At 7 weeks, these performances tended to recover, but in Trail Making A and Figure Position, performances of FH+ alcoholics remained worse than those of controls, while those of FH- alcoholics did not. Thus cognitive recovery during the subacute withdrawal period was worse among FH+ alcoholics than FH- alcoholics, and this finding should be considered when planning alcohol rehabilitation programs. PMID- 16472364 TI - Suicide in Thailand during the period 1998-2003. AB - The aim of this study was to examine the characteristic features of suicides in Thailand between 1998 and 2003. Collected data during 1998-2003 from the Bureau of Policy and Strategy, Ministry of Public Health were analyzed to reveal the mortality from suicide according to age, gender, rate and methods of suicides. Suicide rates were found to have increased to a peak of 8.6 per 100 000 (5290 suicides) in 1999 and then to have decreased to 7.1 per 100 000 in 2003. The average suicide rate during 1998-2003 was 7.9 per 100 000 with a male to female ratio of 3.4:1. Male suicide reached a peak for those aged 25-29 years (21.9 per 100 000) while female suicide showed less variation with age. Hanging was the most common method used, followed by ingestion of agricultural toxic substances. Suicide was most prevalent in upper northern region where HIV infection might be related to the high prevalence. Suicide prevention program should focus on males in early adulthood, and particular measures should be conducted to reduce risk factors related to HIV infection among people in northern Thailand. PMID- 16472365 TI - Putative temperament of patients with generalized anxiety disorder: two-years' interval test-retest reliability of a Japanese version of the Generalized Anxious Temperament. AB - This study examined long-interval test-retest reliability of the Generalized Anxious Temperament (GAT) Japanese version, which is a self-rated questionnaire to measure the temperament of patients with generalized anxiety disorder (GAD). First, we investigated the prevalence of GAT in 214 healthy Japanese subjects (107 females and 107 males). Second, we administered GAT to the 117 (77 females and 40 males) patients with GAD, together with GAF (Global Assessment of Functioning) and HAM-A (Hamilton Anxiety Scale), and readministered after 2 years to examine its reliability. GAT showed good test-retest reliability (Pearson's correlation coefficient = 0.8). Cronbach's alpha of the 26 GAT items was 0.93 that indicated a high level of internal consistency. GAT scores were not influenced by psychological, social or occupational functioning, nor by anxiety symptoms and remained stable over 2 years. GAT scores did not correlate with the age or sex of the control subjects. GAT scores were higher in patients with GAD than they were in control subjects, particularly in females. Twenty-three of 26 GAT items were significantly higher in patients with GAD than in healthy subjects. Stepwise multiple regression analysis revealed that GAT scores in patients with GAD were higher in females than in males (P < 0.01) and in those with early onset of the disorder (P < 0.05). These results suggest that the anxious temperament appears to increase the risk of early onset of GAD, particularly in females. This long interval and large sample research of test retest reliability on GAT indicated its usefulness for understanding the core features of GAD. PMID- 16472366 TI - Group A beta hemolytic streptococcal infections and obsessive-compulsive symptoms in a Turkish pediatric population. AB - The purpose of the present study was to evaluate obsessive-compulsive disease (OCD) in Turkish Children who had group A beta hemolytic streptococcal (GABHS) infections and those who had not. Thirty-one children and adolescents (the study group) were compared with 28 children and adolescents. The Children's Yale-Brown Obsessive-Compulsive Scale (CY-BOCS) scores were rated between study group and control group. The mean score, obsession and compulsion scores of CY-BOCS in the study group were significantly higher than they were in the control group (P < 0.05). The GABHS infections should be assessed in the etiology of OCD in children. Considering GABHS infections may help the treatment of OCD. PMID- 16472367 TI - Beneficial effects of perospirone on aggressive behavior associated with dementia. AB - The aim of this study was to assess the efficacy of the serotonin-dopamine antagonist perospirone in treating aggressive and agitated behavior in patients with dementia. Six patients were referred to the outpatient clinic of Ishizaki Hospital and were followed for 6 weeks. Their psychiatric diagnoses were made using the DSM-IV. Their behavioral symptoms and degrees of cognitive impairment were measured using the Behavioral Pathology in Alzheimer's Disease (BEHAVE-AD) and the Mini-Mental Examination State. The changes in BEHAVE-AD scores were investigated. Maximum benefit was achieved at a mean perospirone dose of 9.0 mg/day. No patient experienced severe adverse effects. Post-hoc analysis showed significant improvement in the total BEHAVE-AD and aggressiveness subscale scores within 2 weeks. This study suggests that perospirone is effective in improving aggressive and agitated behavioral symptoms in demented patients and is safe to use in elderly patients. PMID- 16472368 TI - Local community intervention through depression screening and group activity for elderly suicide prevention. AB - This study aims to evaluate outcomes of a community-based program to prevent suicide among the elderly (>or=65 years old) using a quasi-experimental design with two neighboring references. During 1999-2004, the program including depression screening and group activity was conducted by the public health nurses in the Minami district (population 1685) of Nagawa town, rural Japan. Pre-post changes in the risk of completing suicide were estimated by the incidence rate ratios (IRR). The risk for Minami's elderly females was reduced by 74% (age adjusted IRR, 0.26; 90% CI, 0.07-0.98) more than the historical trend, while there was no change in the risk of Minami's males and nor in the male or female references. The local intervention using public health nursing would be effective against suicide for elderly females without diffusing to the surroundings. PMID- 16472369 TI - Risperidone alternative for a schizophrenic patient with olanzapine-exacerbated diabetic mellitus. PMID- 16472370 TI - Mania after vascular dementia in a patient with bipolar II disorder. PMID- 16472371 TI - Autocastration and penile reconstruction in a patient with obsessive-compulsive disorder. PMID- 16472372 TI - The counterbalance between leptin and cortisol may be associated with comorbid depression and anxiety. PMID- 16472373 TI - Venlafaxine-associated serotonin syndrome and manic episode in a geriatric depressive patient. PMID- 16472374 TI - Donepezil-induced nightmares in mild cognitive impairment. PMID- 16472375 TI - The complete chloroplast DNA sequence of the green alga Oltmannsiellopsis viridis reveals a distinctive quadripartite architecture in the chloroplast genome of early diverging ulvophytes. AB - BACKGROUND: The phylum Chlorophyta contains the majority of the green algae and is divided into four classes. The basal position of the Prasinophyceae has been well documented, but the divergence order of the Ulvophyceae, Trebouxiophyceae and Chlorophyceae is currently debated. The four complete chloroplast DNA (cpDNA) sequences presently available for representatives of these classes have revealed extensive variability in overall structure, gene content, intron composition and gene order. The chloroplast genome of Pseudendoclonium (Ulvophyceae), in particular, is characterized by an atypical quadripartite architecture that deviates from the ancestral type by a large inverted repeat (IR) featuring an inverted rRNA operon and a small single-copy (SSC) region containing 14 genes normally found in the large single-copy (LSC) region. To gain insights into the nature of the events that led to the reorganization of the chloroplast genome in the Ulvophyceae, we have determined the complete cpDNA sequence of Oltmannsiellopsis viridis, a representative of a distinct, early diverging lineage. RESULTS: The 151,933 bp IR-containing genome of Oltmannsiellopsis differs considerably from Pseudendoclonium and other chlorophyte cpDNAs in intron content and gene order, but shares close similarities with its ulvophyte homologue at the levels of quadripartite architecture, gene content and gene density. Oltmannsiellopsis cpDNA encodes 105 genes, contains five group I introns, and features many short dispersed repeats. As in Pseudendoclonium cpDNA, the rRNA genes in the IR are transcribed toward the single copy region featuring the genes typically found in the ancestral LSC region, and the opposite single copy region harbours genes characteristic of both the ancestral SSC and LSC regions. The 52 genes that were transferred from the ancestral LSC to SSC region include 12 of those observed in Pseudendoclonium cpDNA. Surprisingly, the overall gene organization of Oltmannsiellopsis cpDNA more closely resembles that of Chlorella (Trebouxiophyceae) cpDNA. CONCLUSION: The chloroplast genome of the last common ancestor of Oltmannsiellopsis and Pseudendoclonium contained a minimum of 108 genes, carried only a few group I introns, and featured a distinctive quadripartite architecture. Numerous changes were experienced by the chloroplast genome in the lineages leading to Oltmannsiellopsis and Pseudendoclonium. Our comparative analyses of chlorophyte cpDNAs support the notion that the Ulvophyceae is sister to the Chlorophyceae. PMID- 16472376 TI - Transcriptome analysis of a cnidarian-dinoflagellate mutualism reveals complex modulation of host gene expression. AB - BACKGROUND: Cnidarian-dinoflagellate intracellular symbioses are one of the most important mutualisms in the marine environment. They form the trophic and structural foundation of coral reef ecosystems, and have played a key role in the evolutionary radiation and biodiversity of cnidarian species. Despite the prevalence of these symbioses, we still know very little about the molecular modulators that initiate, regulate, and maintain the interaction between these two different biological entities. In this study, we conducted a comparative host anemone transcriptome analysis using a cDNA microarray platform to identify genes involved in cnidarian-algal symbiosis. RESULTS: We detected statistically significant differences in host gene expression profiles between sea anemones (Anthopleura elegantissima) in a symbiotic and non-symbiotic state. The group of genes, whose expression is altered, is diverse, suggesting that the molecular regulation of the symbiosis is governed by changes in multiple cellular processes. In the context of cnidarian-dinoflagellate symbioses, we discuss pivotal host gene expression changes involved in lipid metabolism, cell adhesion, cell proliferation, apoptosis, and oxidative stress. CONCLUSION: Our data do not support the existence of symbiosis-specific genes involved in controlling and regulating the symbiosis. Instead, it appears that the symbiosis is maintained by altering expression of existing genes involved in vital cellular processes. Specifically, the finding of key genes involved in cell cycle progression and apoptosis have led us to hypothesize that a suppression of apoptosis, together with a deregulation of the host cell cycle, create a platform that might be necessary for symbiont and/or symbiont-containing host cell survival. This first comprehensive molecular examination of the cnidarian-dinoflagellate associations provides critical insights into the maintenance and regulation of the symbiosis. PMID- 16472377 TI - Models and partition of variance for quantitative trait loci with epistasis and linkage disequilibrium. AB - BACKGROUND: A genetic model about quantitative trait loci (QTL) provides a basis to interpret the genetic basis of quantitative traits in a study population, such as additive, dominance and epistatic effects of QTL and the partition of genetic variance. The standard quantitative genetics model is based on the least squares partition of genetic effects and also genetic variance in an equilibrium population. However, over years many specialized QTL models have also been proposed for applications in some specific populations. How are these models related? How to analyze and partition a QTL model and genetic variance when both epistasis and linkage disequilibrium are considered? RESULTS: Starting from the classical description of Cockerham genetic model, we first represent the model in a multiple regression setting by using indicator variables to describe the segregation of QTL alleles. In this setting, the definition of additive, dominance and epistatic effects of QTL and the basis for the partition of genetic variance are elaborated. We then build the connection between this general genetic model and a few specialized models (a haploid model, a diploid F2 model and a general two-allele model), and derive the genetic effects and partition of genetic variance for multiple QTL with epistasis and linkage disequilibrium for these specialized models. CONCLUSION: In this paper, we study extensively the composition and property of the genetic model parameters, such as genetic effects and partition of genetic variance, when both epistasis and linkage disequilibrium are considered. This is the first time that both epistasis and linkage disequilibrium are considered in modeling multiple QTL. This analysis would help us to understand the structure of genetic parameters and relationship of various genetic quantities, such as allelic frequencies and linkage disequilibrium, on the definition of genetic effects, and will also help us to understand and properly interpret estimates of the genetic effects and variance components in a QTL mapping experiment. PMID- 16472378 TI - Coarctation of the aorta and mild to moderate developmental delay in a child with a de novo deletion of chromosome 15(q21.1q22.2). AB - BACKGROUND: Deletion of 15q21q22 is a rare chromosomal anomaly. To date, there have been nine reports describing ten individuals with different segmental losses involving 15q21 and 15q22. Many of these individuals have common features of growth retardation, hypotonia and moderate to severe mental retardation. Congenital heart disease has been described in three individuals with interstitial deletion involving this region of chromosome 15. CASE PRESENTATION: We report a child with coarctation of the aorta, partial agenesis of corpus callosum and mild to moderate developmental delay, with a de novo deletion of 15q21.1q22.2, detected by the array Comparative Genomic Hybridization (CGH). We utilized chromosome 15-specific microarray-based CGH to define the chromosomal breakpoints in this patient. CONCLUSION: This is the first description of mapping of an interstitial deletion involving the chromosome 15q21q22 segment using the chromosome 15-specific array-CGH. The report also expands the spectrum of clinical phenotype associated with 15q21q22 deletion. PMID- 16472379 TI - Comparison of the effectiveness of three manual physical therapy techniques in a subgroup of patients with low back pain who satisfy a clinical prediction rule: study protocol of a randomized clinical trial [NCT00257998]. AB - BACKGROUND: Recently a clinical prediction rule (CPR) has been developed and validated that accurately identifies patients with low back pain (LBP) that are likely to benefit from a lumbo-pelvic thrust manipulation. The studies that developed and validated the rule used the identical manipulation procedure. However, recent evidence suggests that different manual therapy techniques may result similar outcomes. The purpose of this study is to investigate the effectiveness of three different manual therapy techniques in a subgroup of patient with low back pain that satisfy the CPR. METHODS/DESIGN: Consecutive patients with LBP referred to physical therapy clinics in one of four geographical locations who satisfy the CPR will be invited to participate in this randomized clinical trial. Subjects who agree to participate will undergo a standard evaluation and complete a number of patient self-report questionnaires including the Oswestry Disability Index (OSW), which will serve as the primary outcome measure. Following the baseline examination patients will be randomly assigned to receive the lumbopelvic manipulation used in the development of the CPR, an alternative lumbar manipulation technique, or non-thrust lumbar mobilization technique for the first 2 visits. Beginning on visit 3, all 3 groups will receive an identical standard exercise program for 3 visits (visits 3,4,5). Outcomes of interest will be captured by a therapist blind to group assignment at 1 week (3rd visit), 4 weeks (6th visit) and at a 6-month follow-up. The primary aim of the study will be tested with analysis of variance (ANOVA) using the change in OSW score from baseline to 4-weeks (OSWBaseline - OSW4-weeks) as the dependent variable. The independent variable will be treatment with three levels (lumbo-pelvic manipulation, alternative lumbar manipulation, lumbar mobilization). DISCUSSION: This trial will be the first to investigate the effectiveness of various manual therapy techniques for patients with LBP who satisfy a CPR. PMID- 16472382 TI - "The case for clean indoor air". PMID- 16472380 TI - A novel expression system of domain I of human beta2 glycoprotein I in Escherichia coli. AB - BACKGROUND: The antiphospholipid syndrome (APS), characterised by recurrent miscarriage and thrombosis, is a significant cause of morbidity and mortality. Domain I (DI) of human beta 2 glycoprotein I (beta2GPI) is thought to contain crucial antibody binding epitopes for antiphospholipid antibodies (aPL), which are critical to the pathogenesis of APS. Expressing this protein in bacteria could facilitate studies investigating how this molecule interacts with aPL. METHODS: Using a computer programme called Juniper, sequentially overlapping primers were designed to be used in a recursive polymerase chain reaction (PCR) to produce a synthetic DI gene. Specifically Juniper incorporates 'major' codons preferred by bacteria altering 41 codons out of 61. This was cloned into the expression plasmid pET(26b) and expressed in BL21(DE3) Escherichia coli (E. coli). By virtue of a pelB leader sequence, periplasmic localisation of DI aided disulphide bond formation and toxicity was addressed by tightly regulating expression through the high stringency T7lac promoter. RESULTS: Purified, soluble his-tagged DI in yields of 750 microg/L bacterial culture was obtained and confirmed on Western blot. Expression using the native human cDNA sequence of DI in the same construct under identical conditions yielded significantly less DI compared to the recombinant optimised sequence. This constitutes the first description of prokaryotic expression of soluble DI of beta2GPI. Binding to murine monoclonal antibodies that recognise conformationally restricted epitopes on the surface of DI and pathogenic human monoclonal IgG aPL was confirmed by direct and indirect immunoassay. Recombinant DI also bound a series of 21 polyclonal IgG samples derived from patients with APS. CONCLUSION: By producing a synthetic gene globally optimised for expression in E. coli, tightly regulating expression and utilising periplasmic product translocation, efficient, soluble E. coli expression of the eukaryotic protein DI of beta2GPI is possible. This novel platform of expression utilising pan-gene prokaryote codon optimisation for DI production will aid future antigenic studies. Furthermore if DI or peptide derivatives of DI are eventually used in the therapeutic setting either as toleragen or as a competitive inhibitor of pathogenic aPL, then an E. coli production system may aid cost-effective production. PMID- 16472383 TI - Rate of promoter class turn-over in yeast evolution. AB - BACKGROUND: Phylogenetic conservation at the DNA level is routinely used as evidence of molecular function, under the assumption that locations and sequences of functional DNA segments remain invariant in evolution. In particular, short DNA segments participating in initiation and regulation of transcription are often conserved between related species. However, transcription of a gene can evolve, and this evolution may involve changes of even such conservative DNA segments. Genes of yeast Saccharomyces have promoters of two classes, class 1 (TATA-containing) and class 2 (non-TATA-containing). RESULTS: Comparison of upstream non-coding regions of orthologous genes from the five species of Saccharomyces sensu stricto group shows that among 212 genes which very likely have class 1 promoters in S. cerevisiae, 17 probably have class 2 promoters in one or more other species. Conversely, among 322 genes which very likely have class 2 promoters in S. cerevisiae, 44 probably have class 1 promoters in one or more other species. Also, for at least 2 genes from the set of 212 S. cerevisiae genes with class 1 promoters, the locations of the TATA consensus sequences are substantially different between the species. CONCLUSION: Our results indicate that, in the course of yeast evolution, a promoter switches its class with the probability at least approximately 0.1 per time required for the accumulation of one nucleotide substitution at a non-coding site. Thus, key sequences involved in initiation of transcription evolve with substantial rates in yeast. PMID- 16472381 TI - Genomic screen for loci associated with tobacco usage in Mission Indians. AB - BACKGROUND: The prevalence of tobacco usage in Native American adults and adolescents is higher than any other racial or ethnic group, yet biological risk and protective factors underlying tobacco use in this ethnic group remain unknown. A genome scan for loci associated with tobacco use phenotypes was performed with data collected from a community sample of Mission Indians residing in Southwest California. METHODS: A structured diagnostic interview was used to define two tobacco use phenotypes: 1) any regular tobacco usage (smoked daily for one month or more) and 2) persistent tobacco usage (smoked at least 10 cigarettes a day for more than one year). Heritability was determined and a linkage analysis was performed, using genotypes for a panel 791 microsatellite polymorphisms, for the two phenotypes using variance component methods implemented in SOLAR. RESULTS: Analyses of multipoint variance component LOD scores for the two tobacco use phenotypes revealed two scores that exceeded 2.0 for the regular use phenotype: one on chromosomes 6 and one on 8. Four other loci on chromosomes 1,7,13, and 22 were found with LOD scores between 1.0 and 1.5. Two loci of interest were found on chromosomes 1 and 4 for the persistent use phenotype with LOD scores between 1.3-1.5. Bivariate linkage analysis was conducted at the site on chromosome 4 for persistent tobacco use and an alcohol drinking severity phenotype previously identified at this site. The maximum LOD score for the bivariate analysis for the region was 3.4, however, there was insufficient power to exclude coincident linkage. CONCLUSION: While not providing evidence for linkage to specific chromosomal regions these results identify regions of interest in the genome in this Mission Indian population, for tobacco usage, some of which were identified in previous genome scans of non-native populations. Additionally, these data lend support for the hypothesis that cigarette smoking, alcohol dependence and other consumptive behaviors may share some common risk and/or protective factors in this Mission Indian population. PMID- 16472384 TI - Thyroid status influence on adiponectin, acylation stimulating protein (ASP) and complement C3 in hyperthyroid and hypothyroid subjects. AB - BACKGROUND: Thyroid abnormalities (hyperthyroid and hypothyroid) are accompanied by changes in intermediary metabolism including alterations in body weight, insulin resistance and lipid profile. The aims of this study were to examine plasma ASP, its precursor C3 and adiponectin in hyperthyroid and hypothyroid subjects as compared to controls. METHODS: A total of 99 subjects were recruited from endocrinology/out-patient clinics: 46 hyperthyroid subjects, 23 hypothyroid subjects and 30 control subjects. Subjects were evaluated for FT4, FT3, TSH, glucose, insulin, complete lipid profile and the adipokines: adiponectin, acylation stimulating protein (ASP) and complement C3. RESULTS: Hyperthyroidism was associated with a 95% increase in adiponectin (p = 0.0002), a 47% decrease in C3 (p < 0.0001), no change in ASP and increased ASP/C3 ratio (p = 0.0012). Hypothyroidism was associated with a 31% increase in ASP (p = 0.008). Adiponectin and C3 correlated with FT3 (r = 0.383, p = 0.004 and r = -0.277, p = 0.007, respectively) and FT4 (r = 0.464, p = 0.003 and r = -0.225, p = 0.03, respectively). ASP correlated with TSH (r = 0.202, p = 0.04). Adiponectin did not correlate with either ASP or C3, only ASP and C3 correlated (r = -0.197, p = 0.05). Adiponectin was negatively correlated with BMI, total cholesterol and plasma triglyceride, while C3 was positively correlated with BMI and total cholesterol. Surprisingly, adiponectin was positively correlated with insulin (r = 0.293, p = 0.02) and HOMA-IR (r = 0.373, p = 0.003) while C3 was negatively correlated with glucose (r = -0.242, p = 0.022, insulin (r = -0.184, p = 0.05) and HOMA-IR. CONCLUSION: These changes suggest that thyroid disease may be accompanied by changes in adipokines, which may contribute to the phenotype expressed. PMID- 16472385 TI - Personal protection of long lasting insecticide-treated nets in areas of Anopheles gambiae s.s. resistance to pyrethroids. AB - BACKGROUND: The development of mosquito nets pre-treated with insecticide, Long Lasting Impregnated Nets (LLINs) that last the life span of the net, is a solution to the difficulty of the re-impregnation of conventional nets. Even if they showed a good efficacy in control conditions, their efficacy in the field, particularly in areas with resistance of Anopheles gambiae to pyrethroids, is not well documented. This study compares wide (Olyset) and small (Permanet) mesh LLINs in field conditions, using entomological parameters. METHODS: The two LLINs were tested in a rice-growing area of south-western Burkina Faso (West Africa) with year around high density of the main malaria vector An. gambiae s.s. In the study village (VK6), there is a mixed population of two molecular forms of An. gambiae, the S-form which dominates during the rainy season and the M-form which dominates the rest of the year. The two LLINs Olyset and Permanet were distributed in the village and 20 matched houses were selected for comparison with four houses without treated nets. RESULTS: Mosquito entrance rate was ten fold higher in control houses than in houses with LLINs and there was no difference between the two net types. Among mosquitoes found in the houses, 36 % were dead in LLIN houses compared to 0% in control houses. Blood feeding rate was 80 % in control houses compared to 43 % in LLIN houses. The type of net did not significantly impact any of these parameters. No mosquitoes were found inside Permanet, whereas dead or dying mosquitoes were collected inside the Olyset. More than 60% of mosquitoes found on top or inside the nets had had blood meals from cattle, as shown by ELISA analysis. CONCLUSION: The percentage of blood-fed mosquitoes in a bed net study does not necessarily determine net success. The efficacy of the two types of LLINs was comparable, during a period when the S form of An. gambiae was carrying the kdr gene. Significantly higher numbers of mosquitoes were collected in control houses compared to intervention houses, indicating that the LLINs provided an additional deterrent effect, which enhanced their expected prevention capacity. PMID- 16472387 TI - Epidemiological trends in nosocomial candidemia in intensive care. AB - BACKGROUND: Infection represents a frequent complication among patients in Intensive Care Units (ICUs) and mortality is high. In particular, the incidence of fungal infections, especially due to Candida spp., has been increasing during the last years. METHODS: In a retrospective study we studied the etiology of candidemia in critically ill patients over a five-year period (1999-2003) in the ICU of the San Martino University Hospital in Genoa, Italy. RESULTS: In total, 182 episodes of candidaemia were identified, with an average incidence of 2.22 episodes/10,000 patient-days/year (range 1.25-3.06 episodes). Incidence of candidemia increased during the study period from 1.25 in 1999 to 3.06/10,000 patient-days/year in 2003. Overall, 40% of the fungemia episodes (74/182) were due to C.albicans, followed by C. parapsilosis (23%), C.glabrata (15%), C.tropicalis (9%) and other species (13%). Candidemia due to non-albicans species increased and this was apparently correlated with an increasing use of azoles for prophylaxis or empirical treatment. CONCLUSION: The study demonstrates a shift in the species of Candida causing fungemia in a medical and surgical ICU population during a 5 year period. The knowledge of the local epidemiological trends in Candida species isolated in blood cultures is important to guide therapeutic choices. PMID- 16472386 TI - Dietary and other lifestyle correlates of serum folate concentrations in a healthy adult population in Crete, Greece: a cross-sectional study. AB - BACKGROUND: Folate has emerged as a key nutrient for optimising health. Impaired folate status has been identified as a risk factor for cardiovascular disease, various types of cancers, and neurocognitive disorders. The study aimed at examining the distribution and determinants of serum folate concentrations in a healthy adult population in Crete, Greece. METHODS: A cross-sectional sample of 486 healthy adults (250 men, 236 women) aged 39 +/- 14 years, personnel of the Medical School and the University Hospital of Crete in Greece, was examined. Serum folate and vitamin B12 concentrations were measured by microbiological assay, and total homocysteine was determined fluorometrically and by high pressure liquid chromatography. Lifestyle questionnaires were completed, and nutrient intakes and food consumption were assessed by 24-h dietary recalls. Multivariate analyses were performed using SPSS v10.1. RESULTS: The geometric mean (95% confidence interval) concentrations of serum folate were 15.6 micromol/l (14.6-16.8) in men and 19.2 micromol/l (17.9-20.7) in women (p < 0.001). Inadequate folate levels ( or = 160 mmHg and/or diastolic blood pressure > or = 95 mmHg) despite the use of antihypertensive drugs, were included. Patients' pharmacy records from 12 months prior to inclusion were obtained. Refill compliance was calculated as the number of days for which the pills were prescribed divided by the total number of days in this period. After inclusion compliance was measured with an electronic monitor that records time and date of each opening of the pillbox. Agreement between both compliance measures was calculated using Spearman's correlation coefficient and Cohen's kappa coefficient. RESULTS: There was very little agreement between the two measures. Whereas refill compliance showed a large range of values, compliance as measured by electronic monitoring was high in almost all patients with estimates between 90% and 100%. Cohen's kappa coefficient was 0.005. CONCLUSION: While electronic monitoring is often considered to be the gold standard for compliance measurements, our results suggest that a short-term electronic monitoring period with the patient being aware of electronic monitoring is probably insufficient to obtain valid compliance data. We conclude that there is a strong need for more studies that explore the effect of electronic monitoring on patient's compliance. PMID- 16472390 TI - Patterns of sexually transmitted infections in adolescents and youth in Dar es Salaam, Tanzania. AB - BACKGROUND: Syndromic management of STIs has been advocated as simplified and cheap approach. Youth have been reported to be at increased risk of acquiring STIs which can facilitate HIV transmission. We have investigated the relationship between the syndromic management and specific aetiology diagnosis and its relationship with HIV infection and health seeking behaviour among youth attending a reproductive health clinic in Dar es Salaam, Tanzania. METHODS: Between September 1998 and February 1999 among 1895 adolescents and youth below 25 years seen in the clinic 199 (10.5%) were randomly selected and consented to participate in the study. A standard questionnaire was administered. Blood and vaginal or urethral specimens were taken and investigated for STI causative agents. RESULTS: Among a total of 199 studied adolescents and youth 22.6 % were teenagers, with fewer females 17.8% than males; 27.5% (p < 0.018). 20.8% of the females compared to 11.5% in males were HIV infected. Genital discharge was the most common complaint which was reported in 54.1% of male and 63.4 % of female patients. All males with gonorrhoea and four out of five with Chlamydia were given appropriate treatment with syndromic management, while 28% women with gonorrhoea or Chlamydia received appropriate treatment by syndromic management. All patients found with active syphilis by serology had not complained of genital ulcers and would not have been assigned to syndromic treatment for syphilis at the initial visit. CONCLUSION: The burden of STIs in this youth population is large indicating that youth are at increased risk of STIs and will certainly require youth friendly clinics. There is a need to refine the current syndromic management guidelines. PMID- 16472392 TI - The pattern of symptoms in patients receiving home based care in Bangwe, Malawi : a descriptive study. AB - BACKGROUND: Home based care of HIV/AIDS patients is a health need recommended but not often available in Africa. Population based assessment helps to identify unmet health needs to plan services. Careful assessment and follow up of patients receiving home based care in a defined population of Bangwe, Malawi provides details of the frequency and severity of common symptoms. METHODS: Mortality and the incidence, duration and severity of common symptoms of patients in a defined population receiving home based care were measured over an eighteen month period. RESULTS: 358 patients, of whom 199 died, were studied. A third of patients died within 4 months of being first seen. About half the patients were unable to care for themselves on first assessment. Half were malnourished with a Body Mass Index (BMI) < 18.5 kg/m2. Most patients had a mixture of symptoms at presentation. These symptoms responded to treatment usually within a fortnight. However a small proportion (5%) of patients suffered repeated episodes often as many as 6-9 times a year. Incidence rates are estimated. CONCLUSION: Symptoms which are alleviated by simple treatments are common. The patients in whom symptoms recur need a responsive home based care service. Population based estimates of incidence and duration of disease and the visit work load allow an assessment of home based care needs in an urban setting in Africa. PMID- 16472391 TI - Analysis of case-parent trios at a locus with a deletion allele: association of GSTM1 with autism. AB - BACKGROUND: Certain loci on the human genome, such as glutathione S-transferase M1 (GSTM1), do not permit heterozygotes to be reliably determined by commonly used methods. Association of such a locus with a disease is therefore generally tested with a case-control design. When subjects have already been ascertained in a case-parent design however, the question arises as to whether the data can still be used to test disease association at such a locus. RESULTS: A likelihood ratio test was constructed that can be used with a case-parents design but has somewhat less power than a Pearson's chi-squared test that uses a case-control design. The test is illustrated on a novel dataset showing a genotype relative risk near 2 for the homozygous GSTM1 deletion genotype and autism. CONCLUSION: Although the case-control design will remain the mainstay for a locus with a deletion, the likelihood ratio test will be useful for such a locus analyzed as part of a larger case-parent study design. The likelihood ratio test has the advantage that it can incorporate complete and incomplete case-parent trios as well as independent cases and controls. Both analyses support (p = 0.046 for the proposed test, p = 0.028 for the case-control analysis) an association of the homozygous GSTM1 deletion genotype with autism. PMID- 16472393 TI - Intracellular assembly and budding of the Murine Leukemia Virus in infected cells. AB - BACKGROUND: Murine Leukemia Virus (MLV) assembly has been long thought to occur exclusively at the plasma membrane. Current models of retroviral particle assembly describe the recruitment of the host vacuolar protein sorting machinery to the cell surface to induce the budding of new particles. Previous fluorescence microscopy study reported the vesicular traffic of the MLV components (Gag, Env and RNA). Here, electron microscopy (EM) associated with immunolabeling approaches were used to go deeply into the assembly of the "prototypic" MLV in chronically infected NIH3T3 cells. RESULTS: Beside the virus budding events seen at the cell surface of infected cells, we observed that intracellular budding events could also occur inside the intracellular vacuoles in which many VLPs accumulated. EM in situ hybridization and immunolabeling analyses confirmed that these latter were MLV particles. Similar intracellular particles were detected in cells expressing MLV Gag alone. Compartments containing the MLV particles were identified as late endosomes using Lamp1 endosomal/lysosomal marker and BSA-gold pulse-chase experiments. In addition, infectious activity was detected in lysates of infected cells. CONCLUSION: Altogether, our results showed that assembly of MLV could occur in part in intracellular compartments of infected murine cells and participate in the production of infectious viruses. These observations suggested that MLV budding could present similarities with the particular intracellular budding of HIV in infected macrophages. PMID- 16472394 TI - The Shoulder Pain and Disability Index demonstrates factor, construct and longitudinal validity. AB - BACKGROUND: The Shoulder Pain and Disability Index (SPADI) is a self-report measure developed to evaluate patients with shoulder pathology. While some validation has been conducted, broader analyses are indicated. This study determined aspects of cross-sectional and longitudinal validity of the SPADI. METHODS: Community volunteers (n = 129) who self-identified as having shoulder pain were enrolled. Patients were examined by a physical therapist using a standardized assessment process to insure that their pain was musculoskeletal in nature. This included examination of pain reported during active and passive shoulder motion as reported on a visual analogue pain scale. Patients completed the SPADI, the Coping Strategies Questionnaire (CSQ) and the Sickness Impact Profile (SIP) at a baseline assessment and again 3 and 6 months later. Factor analysis with varimax rotation was used to assess subscale structure. Expectations regarding convergent and divergent subscales of CSQ and SIP were determined a priori and analysed using Pearson correlations. Constructed hypotheses that patients with a specific diagnosis or on pain medication would demonstrate higher SPADI scores were tested. Correlations between the observed changes recorded across different instruments were used to assess longitudinal validity. RESULTS: The internal consistencies of the SPADI subscales were high (alpha > 0.92). Factor analysis with varimax rotation indicated that the majority of items fell into 2 factors that represent pain and disability. Two difficult functional items tended to align with pain items. Higher pain and disability was correlated to passive or negative coping strategies, i.e., praying/hoping, catastrophizing on the CSQ. The correlations between subscales of the SPADI and SIP were low with divergent subscales and low to moderate with convergent subscales. Correlations, r > 0.60, were observed between the SPADI and pain reported on a VAS pain scale during active and passive movement. The two constructed validity hypotheses (on diagnosis and use of pain medications) were both supported (p < 0.01). The SPADI demonstrated significant changes over time, but these were poorly correlated to the SIP or CSQ suggesting that these scales measure different parameters. CONCLUSION: The SPADI is a valid measure to assess pain and disability in community-based patients reporting shoulder pain due to musculoskeletal pathology. PMID- 16472395 TI - Uteroplacental bleeding disorders during pregnancy: do missing paternal characteristics influence risk? AB - BACKGROUND: Several studies have assessed the risks of uteroplacental bleeding disorders in relation to maternal characteristics. The association between uteroplacental bleeding disorders and paternal characteristics, however, has received considerably less attention. Data on paternal demographics, notably race and age, from birth certificate data are becoming increasingly incomplete in recent years. This pattern of increasingly underreporting of paternal demographic data led us to speculate that pregnancies for which paternal characteristics are partially or completely missing may be associated with increased risk for uteroplacental bleeding disorders. The objective of this study is to examine the association between placenta previa and placental abruption and missing paternal age and race. METHODS: A retrospective cohort study using U.S. linked birth/infant death data from 1995 through 2001 (n = 26,336,549) was performed. Risks of placenta previa and placental abruption among: (i) pregnancies with complete paternal age and race data; (ii) paternal age only missing; (iii) paternal race only missing; and (iv) both paternal age and race missing, were evaluated. Relative risk (RR) with 95% confidence interval (CI) for placenta previa and placental abruption by missing paternal characteristics were derived after adjusting for confounders. RESULTS: Adjusted RR for placental abruption were 1.30 (95% CI 1.24, 1.37), 1.00 (95% CI 0.95, 1.05), and 1.08 (95% CI 1.06, 1.10) among pregnancies with "paternal age only", "paternal race only", and "both paternal age and race" missing, respectively. The increased risk of placental abruption among the "paternal age only missing" category is partly explained by increased risks among whites aged 20-29 years, and among blacks aged >or=30 years. However, no clear patterns in the associations between missing paternal characteristics and placenta previa were evident. CONCLUSION: Missing paternal characteristics are associated with increased risk of placental abruption, likely mediated through low socio-economic conditions. PMID- 16472396 TI - A multiscale mathematical model of cancer, and its use in analyzing irradiation therapies. AB - BACKGROUND: Radiotherapy outcomes are usually predicted using the Linear Quadratic model. However, this model does not integrate complex features of tumor growth, in particular cell cycle regulation. METHODS: In this paper, we propose a multiscale model of cancer growth based on the genetic and molecular features of the evolution of colorectal cancer. The model includes key genes, cellular kinetics, tissue dynamics, macroscopic tumor evolution and radiosensitivity dependence on the cell cycle phase. We investigate the role of gene-dependent cell cycle regulation in the response of tumors to therapeutic irradiation protocols. RESULTS: Simulation results emphasize the importance of tumor tissue features and the need to consider regulating factors such as hypoxia, as well as tumor geometry and tissue dynamics, in predicting and improving radiotherapeutic efficacy. CONCLUSION: This model provides insight into the coupling of complex biological processes, which leads to a better understanding of oncogenesis. This will hopefully lead to improved irradiation therapy. PMID- 16472397 TI - Detection of Chlamydia in the peripheral blood cells of normal donors using in vitro culture, immunofluorescence microscopy and flow cytometry techniques. AB - BACKGROUND: Chlamydia trachomatis (Ct) and Chlamydia pneumoniae (Cp) are medically significant infectious agents associated with various chronic human pathologies. Nevertheless, specific roles in disease progression or initiation are incompletely defined. Both pathogens infect established cell lines in vitro and polymerase chain reaction (PCR) has detected Chlamydia DNA in various clinical specimens as well as in normal donor peripheral blood monocytes (PBMC). However, Chlamydia infection of other blood cell types, quantification of Chlamydia infected cells in peripheral blood and transmission of this infection in vitro have not been examined. METHODS: Cp specific titers were assessed for sera from 459 normal human donor blood (NBD) samples. Isolated white blood cells (WBC) were assayed by in vitro culture to evaluate infection transmission of blood cell borne chlamydiae. Smears of fresh blood samples (FB) were dual immunostained for microscopic identification of Chlamydia-infected cell types and aliquots also assessed using Flow Cytometry (FC). RESULTS: ELISA demonstrated that 219 (47.7%) of the NBD samples exhibit elevated anti-Cp antibody titers. Imunofluorescence microscopy of smears demonstrated 113 (24.6%) of samples contained intracellular Chlamydia and monoclonals to specific CD markers showed that in vivo infection of neutrophil and eosinophil/basophil cells as well as monocytes occurs. In vitro culture established WBCs of 114 (24.8%) of the NBD samples harbored infectious chlamydiae, clinically a potentially source of transmission, FC demonstrated both Chlamydia infected and uninfected cells can be readily identified and quantified. CONCLUSION: NBD can harbor infected neutrophils, eosinophil/basophils and monocytes. The chlamydiae are infectious in vitro, and both total, and cell type specific Chlamydia carriage is quantifiable by FC. PMID- 16472398 TI - Horizontal gene transfer from Bacteria to rumen Ciliates indicates adaptation to their anaerobic, carbohydrates-rich environment. AB - BACKGROUND: The horizontal transfer of expressed genes from Bacteria into Ciliates which live in close contact with each other in the rumen (the foregut of ruminants) was studied using ciliate Expressed Sequence Tags (ESTs). More than 4000 ESTs were sequenced from representatives of the two major groups of rumen Cilates: the order Entodiniomorphida (Entodinium simplex, Entodinium caudatum, Eudiplodinium maggii, Metadinium medium, Diploplastron affine, Polyplastron multivesiculatum and Epidinium ecaudatum) and the order Vestibuliferida, previously called Holotricha (Isotricha prostoma, Isotricha intestinalis and Dasytricha ruminantium). RESULTS: A comparison of the sequences with the completely sequenced genomes of Eukaryotes and Prokaryotes, followed by large scale construction and analysis of phylogenies, identified 148 ciliate genes that specifically cluster with genes from the Bacteria and Archaea. The phylogenetic clustering with bacterial genes, coupled with the absence of close relatives of these genes in the Ciliate Tetrahymena thermophila, indicates that they have been acquired via Horizontal Gene Transfer (HGT) after the colonization of the gut by the rumen Ciliates. CONCLUSION: Among the HGT candidates, we found an over representation (>75%) of genes involved in metabolism, specifically in the catabolism of complex carbohydrates, a rich food source in the rumen. We propose that the acquisition of these genes has greatly facilitated the Ciliates' colonization of the rumen providing evidence for the role of HGT in the adaptation to new niches. PMID- 16472399 TI - Older adults' beliefs about physician-estimated life expectancy: a cross sectional survey. AB - BACKGROUND: Estimates of life expectancy assist physicians and patients in medical decision-making. The time-delayed benefits for many medical treatments make an older adult's life expectancy estimate particularly important for physicians. The purpose of this study is to assess older adults' beliefs about physician-estimated life expectancy. METHODS: We performed a mixed qualitative quantitative cross-sectional study in which 116 healthy adults aged 70+ were recruited from two local retirement communities. We interviewed them regarding their beliefs about physician-estimated life expectancy in the context of a larger study on cancer screening beliefs. Semi-structured interviews of 80 minutes average duration were performed in private locations convenient to participants. Demographic characteristics as well as cancer screening beliefs and beliefs about life expectancy were measured. Two independent researchers reviewed the open-ended responses and recorded the most common themes. The research team resolved disagreements by consensus. RESULTS: This article reports the life expectancy results portion of the larger study. The study group (n = 116) was comprised of healthy, well-educated older adults, with almost a third over 85 years old, and none meeting criteria for dementia. Sixty-four percent (n = 73) felt that their physicians could not correctly estimate their life expectancy. Sixty-six percent (n = 75) wanted their physicians to talk with them about their life expectancy. The themes that emerged from our study indicate that discussions of life expectancy could help older adults plan for the future, maintain open communication with their physicians, and provide them knowledge about their medical conditions. CONCLUSION: The majority of the healthy older adults in this study were open to discussions about life expectancy in the context of discussing cancer screening tests, despite awareness that their physicians' estimates could be inaccurate. Since about a third of participants perceived these discussions as not useful or even harmful, physicians should first ascertain patients' preferences before discussing their life expectancies. PMID- 16472401 TI - Comparison between two methods of working length determination and its effect on radiographic extent of root canal filling: a clinical study [ISRCTN71486641]. AB - BACKGROUND: Obtaining a correct working length is critical to the success of endodontic therapy. Different methods have been used to identify this crucial measurement. The Aim of this clinical study was to compare the effect of working length determination using apex locator alone or in combination with working length radiograph on the apical extent of root canal filling. METHODS: A total number of 66 patients, 151 canals were randomized into two groups, In group (I) working length was determined by apex locator alone, while in group (II) working length was determined by apex locator confirmed by working length radiograph, length of obturation was assessed, and the total number of radiographs was recorded. The data were analyzed using SAS system and T. tests were carried out. Statistical significance was considered to be P 0.05). CONCLUSION: The practice of using electronic apex locator in the determination of working length is useful and reliable with no statistical difference of the radiographic extent of root canal filling when using apex locator alone or in combination with working length radiograph. Under the clinical conditions of this study, it is suggested that the correct use of an apex locator alone could prevent the need for further diagnostic radiographs for determination of working length. This method can be useful in patients who need not to be exposed to repeated radiation because of mental, medical or oral conditions. PMID- 16472400 TI - Phylogenetic identification of lateral genetic transfer events. AB - BACKGROUND: Lateral genetic transfer can lead to disagreements among phylogenetic trees comprising sequences from the same set of taxa. Where topological discordance is thought to have arisen through genetic transfer events, tree comparisons can be used to identify the lineages that may have shared genetic information. An 'edit path' of one or more transfer events can be represented with a series of subtree prune and regraft (SPR) operations, but finding the optimal such set of operations is NP-hard for comparisons between rooted trees, and may be so for unrooted trees as well. RESULTS: Efficient Evaluation of Edit Paths (EEEP) is a new tree comparison algorithm that uses evolutionarily reasonable constraints to identify and eliminate many unproductive search avenues, reducing the time required to solve many edit path problems. The performance of EEEP compares favourably to that of other algorithms when applied to strictly bifurcating trees with specified numbers of SPR operations. We also used EEEP to recover edit paths from over 19,000 unrooted, incompletely resolved protein trees containing up to 144 taxa as part of a large phylogenomic study. While inferred protein trees were far more similar to a reference supertree than random trees were to each other, the phylogenetic distance spanned by random versus inferred transfer events was similar, suggesting that real transfer events occur most frequently between closely related organisms, but can span large phylogenetic distances as well. While most of the protein trees examined here were very similar to the reference supertree, requiring zero or one edit operations for reconciliation, some trees implied up to 40 transfer events within a single orthologous set of proteins. CONCLUSION: Since sequence trees typically have no implied root and may contain unresolved or multifurcating nodes, the strategy implemented in EEEP is the most appropriate for phylogenomic analyses. The high degree of consistency among inferred protein trees shows that vertical inheritance is the dominant pattern of evolution, at least for the set of organisms considered here. However, the edit paths inferred using EEEP suggest an important role for genetic transfer in the evolution of microbial genomes as well. PMID- 16472402 TI - Computational models with thermodynamic and composition features improve siRNA design. AB - BACKGROUND: Small interfering RNAs (siRNAs) have become an important tool in cell and molecular biology. Reliable design of siRNA molecules is essential for the needs of large functional genomics projects. RESULTS: To improve the design of efficient siRNA molecules, we performed a comparative, thermodynamic and correlation analysis on a heterogeneous set of 653 siRNAs collected from the literature. We used this training set to select siRNA features and optimize computational models. We identified 18 parameters that correlate significantly with silencing efficiency. Some of these parameters characterize only the siRNA sequence, while others involve the whole mRNA. Most importantly, we derived an siRNA position-dependent consensus, and optimized the free-energy difference of the 5' and 3' terminal dinucleotides of the siRNA antisense strand. The position dependent consensus is based on correlation and t-test analyses of the training set, and accounts for both significantly preferred and avoided nucleotides in all sequence positions. On the training set, the two parameters' correlation with silencing efficiency was 0.5 and 0.36, respectively. Among other features, a dinucleotide content index and the frequency of potential targets for siRNA in the mRNA added predictive power to our model (R = 0.55). We showed that our model is effective for predicting the efficiency of siRNAs at different concentrations. We optimized a neural network model on our training set using three parameters characterizing the siRNA sequence, and predicted efficiencies for the test siRNA dataset recently published by Novartis. On this validation set, the correlation coefficient between predicted and observed efficiency was 0.75. Using the same model, we performed a transcriptome-wide analysis of optimal siRNA targets for 22,600 human mRNAs. CONCLUSION: We demonstrated that the properties of the siRNAs themselves are essential for efficient RNA interference. The 5' ends of antisense strands of efficient siRNAs are U-rich and possess a content similarity to the pyrimidine-rich oligonucleotides interacting with the polypurine RNA tracks that are recognized by RNase H. The advantage of our method over similar methods is the small number of parameters. As a result, our method requires a much smaller training set to produce consistent results. Other mRNA features, though expensive to compute, can slightly improve our model. PMID- 16472403 TI - An analysis of sickness absence in chronically ill patients receiving complementary and alternative medicine: a longterm prospective intermittent study. AB - BACKGROUND: The popularity of complementary and alternative medicine (CAM) has led to a growing amount of research in this area. All the same little is known about the effects of these special treatments in every-day practice of primary care, delivered by general practitioners within the health insurance system. From 1994 to 2000 more than 20 German Company health insurances initiated the first model project on CAM according to the German social law. Aim of this contribution is to investigate the effectiveness of multi-modal CAM on chronic diseases within primary health care. METHODS: A long-term prospective intermittent study was conducted including 44 CAM practitioners and 1221 self-selected chronically ill patients (64% women) of whom 441 were employed. Main outcome measure is sick leave, controlled for secular trends and regression-to-the mean and self perceived health status. RESULTS: Sick-leave per year of 441 patients at work increased from 22 (SD +/- 45.2) to 31 (+/- 61.0) days within three years prior to intervention, and decreased to 24 (+/- 55.6) in the second year of treatment, sustaining at this level in the following two years. Detailed statistical analysis show that this development exceeds secular trends and the regression toward-the-mean effect. Sick-leave reduction was corroborated by data on self reported improvement of patients' health status. CONCLUSION: Results of this longterm observational study show a reduction of sick leave in chronically ill patients after a complex multimodal CAM intervention. However, as this is an uncontrolled observational study efficacy of any specific CAM treatment can not be proven. The results might indicate an general effectiveness of CAM in primary care, worthwhile further investigations. Future studies should identify the most suitable patients for CAM practices, the most appropriate and safe treatments, provide information on the magnitude of the effects to facilitate subsequent definitive randomised controlled studies that will help to position complementary and alternative medicine in health care. PMID- 16472404 TI - Prolonged ozone exposure in an allergic airway disease model: adaptation of airway responsiveness and airway remodeling. AB - BACKGROUND: Short-term exposure to high concentrations of ozone has been shown to increase airway hyper-responsiveness (AHR). Because the changes in AHR and airway inflammation and structure after chronic ozone exposure need to be determined, the goal of this study was to investigate these effects in a murine model of allergic airway disease. METHODS: We exposed BALB/c mice to 2 ppm ozone for 4, 8, and 12 weeks. We measured the enhanced pause (Penh) to methacholine and performed cell differentials in bronchoalveolar lavage fluid. We quantified the levels of IL-4 and IFN-gamma in the supernatants of the bronchoalveolar lavage fluids using enzyme immunoassays, and examined the airway architecture under light and electron microscopy. RESULTS: The groups exposed to ozone for 4, 8, and 12 weeks demonstrated decreased Penh at methacholine concentrations of 12.5, 25, and 50 mg/ml, with a dose-response curve to the right of that for the filtered-air group. Neutrophils and eosinophils increased in the group exposed to ozone for 4 weeks compared to those in the filtered-air group. The ratio of IL-4 to INF-gamma increased significantly after exposure to ozone for 8 and 12 weeks compared to the ratio for the filtered-air group. The numbers of goblet cells, myofibroblasts, and smooth muscle cells showed time-dependent increases in lung tissue sections from the groups exposed to ozone for 4, 8, and 12 weeks. CONCLUSION: These findings demonstrate that the increase in AHR associated with the allergic airway does not persist during chronic ozone exposure, indicating that airway remodeling and adaptation following repeated exposure to air pollutants can provide protection against AHR. PMID- 16472405 TI - Molecular characterization and differential expression of cytokinin-responsive type-A response regulators in rice (Oryza sativa). AB - BACKGROUND: The response regulators represent the elements of bacterial two component system and have been characterized from dicot plants like Arabidopsis but little information is available on the monocots, including the cereal crops. The aim of this study was to characterize type-A response regulator genes from rice, and to investigate their expression in various organs as well as in response to different hormones, including cytokinin, and environmental stimuli. RESULTS: By analysis of the whole genome sequence of rice, we have identified ten genes encoding type-A response regulators based upon their high sequence identity within the receiver domain. The exon-intron organization, intron-phasing as well as chromosomal location of all the RT-PCR amplified rice (Oryza sativa) response regulator (OsRR) genes have been analyzed. The transcripts of OsRR genes could be detected by real-time PCR in all organs of the light- and dark-grown rice seedlings/plants, although there were quantitative differences. The steady-state transcript levels of most of the OsRR genes increased rapidly (within 15 min) on exogenous cytokinin application even in the presence of cycloheximide. Moreover, the expression of the OsRR6 gene was enhanced in rice seedlings exposed to salinity, dehydration and low temperature stress. CONCLUSION: Ten type-A response regulator genes identified in rice, the model monocot plant, show overlapping/differential expression patterns in various organs and in response to light. The induction of OsRR genes by cytokinin even in the absence of de novo protein synthesis qualifies them to be primary cytokinin response genes. The induction of OsRR6 in response to different environmental stimuli indicates its role in cross-talk between abiotic stress and cytokinin signaling. These results provide a foundation for further investigations on specific as well as overlapping cellular functions of type-A response regulators in rice. PMID- 16472407 TI - Quantification of the fractal nature of mycelial aggregation in Aspergillus niger submerged cultures. AB - BACKGROUND: Fractal geometry estimates have proven useful in studying the growth strategies of fungi in response to different environments on soil or on agar substrates, but their use in mycelia grown submerged is still rare. In the present study, the effects of certain important fermentation parameters, such as the spore inoculum level, phosphate and manganese concentrations in the medium, on mycelial morphology of the citric acid producer Aspergillus niger were determined by fractal geometry. The value of employing fractal geometry to describe mycelial structures was examined in comparison with information from other descriptors including classic morphological parameters derived from image analysis. RESULTS: Fractal analysis of distinct morphological forms produced by fermentation conditions that influence fungal morphology and acid production, showed that the two fractal dimensions DBS (box surface dimension) and DBM (box mass dimension) are very sensitive indexes, capable of describing morphological differences. The two box-counting methods applied (one applied to the whole mass of the mycelial particles and the other applied to their surface only) enabled evaluation of fractal dimensions for mycelial particles in this analysis in the region of DBS = 1.20-1.70 and DBM = 1.20-2.70. The global structure of sufficiently branched mycelia was described by a single fractal dimension D, which did not exceed 1.30. Such simple structures are true mass fractals (DBS = DBM = D) and they could be young mycelia or dispersed forms of growth produced by very dense spore inocula (108-109 spores/ml) or by addition of manganese in the medium. Mycelial clumps and pellets were effectively discriminated by fractal analysis. Fractal dimension values were plotted together with classic morphological parameters derived from image analysis for comparisons. Their sensitivity to treatment was analogous to the sensitivity of classic morphological parameters suggesting that they could be equally used as morphological descriptors. CONCLUSION: Starting from a spore, the mycelium develops as a mass fractal and, depending on culture conditions, it either turns to a surface fractal or remains a mass fractal. Since fractal dimensions give a measure of the degree of complexity and the mass filling properties of an object, it may be possible that a large number of morphological parameters which contribute to the overall complexity of the particles, could be replaced by these indexes effectively. PMID- 16472406 TI - Total plasma homocysteine, folate, and vitamin B12 status in healthy Iranian adults: the Tehran homocysteine survey (2003-2004)/a cross-sectional population based study. AB - BACKGROUND: Elevated plasma total homocysteine is an independent risk factor for cardiovascular disease and a sensitive marker of the inadequate vitamin B12 and folate insufficiency. Folate and vitamin B12 have a protective effect on cardiovascular disease. This population based study was conducted to evaluate the plasma total homocysteine, folate, and vitamin B12 in healthy Iranian individuals. METHODS: This study was a part of the Cardiovascular Risk Factors Survey in the Population Lab Region of Tehran University has been designed and conducted based on the methodology of MONICA/WHO Project. A total of 1214 people aged 25-64 years, were recruited and assessed regarding demographic characteristics, homocysteine, folate, and vitamin B12 levels with interview, questionnaires, examination and blood sampling. Blood samples were gathered and analyzed according to standard methods. RESULTS: The variables were assessed in 1214 participants including 428 men (35.3%) and 786 women (64.7%). Age-adjusted prevalence of hyperhomocysteinemia (Hcy > or = 15 micromol/L) was 73.1% in men and 41.07% in women (P < 0.0001). Geometric mean of plasma homocysteine was 19.02 +/- 1.46 micromol/l in men and 14.05 +/- 1.45 micromol/l in women (P < 0.004) which increased by ageing. Age-adjusted prevalence of low serum folate level was 98.67% in men and 97.92% in women. Age-adjusted prevalence of low serum vitamin B12 level was 26.32% in men and 27.2% in women. Correlation coefficients (Pearson's r) between log tHcy and serum folate, and vitamin B12 indicated an inverse correlation (r = -0.27, r = -0.19, P < 0.0001, respectively). CONCLUSION: These results revealed that the prevalence of hyperhomocysteinemia, low folate and vitamin B12 levels are considerably higher than other communities. Implementation of preventive interventions such as food fortification with folic acid is necessary. PMID- 16472408 TI - Attentional processes discriminate between patients with mild Alzheimer's disease and cognitively healthy elderly. AB - BACKGROUND: It is generally accepted that Alzheimer's disease (AD) is mainly characterized by memory disorders. Although recent studies also point to an important role of attention deficits early in the disease, this notion has not yet emerged in clinical practice. Our aim was to assess whether attention, quantified by reaction times, can discriminate between patients with mild AD and controls and therefore contribute to clinical diagnosis. METHODS: In a cross sectional study, 33 patients with mild AD were matched with cognitively healthy elderly controls for age, gender, educational level and depressive mood. Selective attention (SA), alternating attention (AA) and error-rates were measured by a modified reaction time test. RESULTS: Significant differences between both groups were found for all measures. Logistic regression showed that SA (corrected for individual processing speed) and error-rates could correctly classify subjects with an overall hit ratio of 81%. When attention measures were not corrected for individual processing speed, the overall hit ratio improved to 97%. CONCLUSION: SA and AA deteriorate in patients with mild AD and these measures can be used to discriminate between patients and matched controls, independently of depressive mood. PMID- 16472409 TI - Behavioral interventions for agitation in older adults with dementia: an evaluative review. AB - BACKGROUND: Older adults with dementia commonly exhibit agitated behavior that puts them at risk of injury and institutionalization and is associated with caregiver stress. A range of theoretical approaches has produced numerous interventions to manage these behavior problems. This paper critically reviews the empirical literature on behavioral interventions to reduce agitation in older adults with dementia. METHOD: A literature search yielded 23 articles that met inclusion criteria. These articles described interventions that targeted wandering, disruptive vocalization, physical aggression, other agitated behaviors and a combination of these behaviors. Studies are summarized individually and then evaluated. RESULTS: Behavioral interventions targeting agitated behavior exhibited by older adults with dementia show considerable promise. A number of methodological issues must be addressed to advance this research area. Problem areas include inconsistent use of functional assessment techniques, failure to report quantitative findings and inadequate demonstrations of experimental control. CONCLUSIONS: The reviewed studies collectively provide evidence that warrants optimism regarding the application of behavioral principles to the management of agitation among older adults with dementia. Although the results of some studies were mixed and several studies revealed methodological shortcomings, many of them offered innovations that can be used in future, more rigorously designed, intervention studies. PMID- 16472410 TI - Lack of association between iron metabolism and depressive mood in an elderly general population. AB - BACKGROUND: Alterations in iron metabolism have been suggested as potential pathological markers in patients with manifest depression. No data on the association between iron and depression exist from population-based studies, in which milder forms of depressive symptoms are much more common. The aim of this study was to analyze the relationship between six parameters of iron metabolism and depressive mood in a population-based cross-sectional study in Germany. METHODS: A total of 374 participants, aged 65-83 years, of the Memory and Morbidity in Augsburg Elderly (MEMO) Study were assessed using the Center for Epidemiologic Studies - Depression Scale (CES-D) for depression. Iron, ferritin, transferrin, soluble transferrin receptor, iron binding capacity, transferrin saturation and C-reactive protein were analyzed with standard laboratory methods. Linear and logistic regression analyses were applied to evaluate the relationship between iron parameters and depressive mood. RESULTS: The 7-day prevalence of depressive mood was 10.2%, with a higher risk in women compared to men [odds ratio (OR) = 2.04; 95% confidence interval (95% CI) = 1.04-4.0]. Correlation and linear regression analyses adjusted for age, gender, hypertension and smoking yielded no significant relationship between any of the iron parameters and the CES-D scores. In gender-stratified analyses a statistically significant association between serum iron and depressive mood was observed in men only. This finding disappeared after applying a Bonferroni correction for multiple testing. CONCLUSIONS: The lack of association of iron metabolism and depressive mood reported in this population-based study does not support previous findings in patients with major depression. This negative finding in milder forms of depression in elderly people indicates either the absence or a more complex nature of the interactions between iron metabolism, low-grade inflammation and depression. PMID- 16472414 TI - Size relationships between the parasitic copepod, Lernanthropus cynoscicola , and its fish host, Cynoscion guatucupa. AB - The effects of the size of Cynoscion guatucupa on the size and demographic parameters of their parasitic copepod Lernanthropus cynoscicola were evaluated. Prevalence of copepods increased with host size up to fish of intermediate length, then it decreased, probably because changes in size of gill filaments affect their attachment capability, enhancing the possibility of being detached by respiratory currents. Body length of copepods was significantly correlated with host length, indicating that only parasites of an 'adequate' size can be securely attached to a fish of a given size. The absence of relationship between the coefficient of variability in copepod length and both host length and number of conspecifics, together with the host-size dependence of both male and juvenile female sizes, prevent to interpret this relationship as a phenomenon of developmental plasticity. Therefore, the observed peak of prevalence could reflect the distribution of size frequencies in the population of copepods, with more individuals near the average length. Concluding, the 'optimum' host size for L. cynoscicola could merely be the adequate size for most individuals in the population, depending, therefore, on a populational attribute of parasites. However, its location along the host size range could be determined by a balance between fecundity and number of available hosts, which increases and decreases, respectively, with both host and parasite size. PMID- 16472413 TI - A classification of tasks for the systematic study of immune response using functional genomics data. AB - A full understanding of the immune system and its responses to infection by different pathogens is important for the development of anti-parasitic vaccines. A growing number of large-scale experimental techniques, such as microarrays, are being used to gain a better understanding of the immune system. To analyse the data generated by these experiments, methods such as clustering are widely used. However, individual applications of these methods tend to analyse the experimental data without taking publicly available biological and immunological knowledge into account systematically and in an unbiased manner. To make best use of the experimental investment, to benefit from existing evidence, and to support the findings in the experimental data, available biological information should be included in the analysis in a systematic manner. In this review we present a classification of tasks that shows how experimental data produced by studies of the immune system can be placed in a broader biological context. Taking into account available evidence, the classification can be used to identify different ways of analysing the experimental data systematically. We have used the classification to identify alternative ways of analysing microarray data, and illustrate its application using studies of immune responses in mice to infection with the intestinal nematode parasites Trichuris muris and Heligmosomoides polygyrus. PMID- 16472412 TI - Triadic model of the neurobiology of motivated behavior in adolescence. AB - BACKGROUND: Risk-taking behavior is a major cause of morbidity and mortality in adolescence. In the context of decision theory and motivated (goal-directed) behavior, risk-taking reflects a pattern of decision-making that favors the selection of courses of action with uncertain and possibly harmful consequences. We present a triadic, neuroscience systems-based model of adolescent decision making. METHOD: We review the functional role and neurodevelopmental findings of three key structures in the control of motivated behavior, i.e. amygdala, nucleus accumbens, and medial/ventral prefrontal cortex. We adopt a cognitive neuroscience approach to motivated behavior that uses a temporal fragmentation of a generic motivated action. Predictions about the relative contributions of the triadic nodes to the three stages of a motivated action during adolescence are proposed. RESULTS: The propensity during adolescence for reward/novelty seeking in the face of uncertainty or potential harm might be explained by a strong reward system (nucleus accumbens), a weak harm-avoidant system (amygdala), and/or an inefficient supervisory system (medial/ventral prefrontal cortex). Perturbations in these systems may contribute to the expression of psychopathology, illustrated here with depression and anxiety. CONCLUSIONS: A triadic model, integrated in a temporally organized map of motivated behavior, can provide a helpful framework that suggests specific hypotheses of neural bases of typical and atypical adolescent behavior. PMID- 16472417 TI - [Renal giant angiomyolipoma in tuberous sclerosis complex: case report and literature review]. AB - The aim of this paper is to describe a typical clinical case of tuberous sclerosis complex (Bourneville disease) and discuss controversial issues about the management of this rare condition, with a short revision of the literature. Particularly, we define which is the role of the surgeon in the treatment of this very rare condition, that should be primary approached conservatively. PMID- 16472416 TI - Medullary thyroid carcinoma: state of the art. AB - Medullary thyroid carcinoma (MTC) constitutes about 3-10% of all thyroid cancers. It arises from the parafollicular C cells that produce calcitonin (CT) and occurs as a sporadic form. or less commonly, as a hereditary form, as part of multiple endocrine neoplasia syndromes types 2A (MEN 2A) and 2B (MEN 2B). The hereditary forms are autosomal dominant traits associated with germline mutations of RET proto-oncogene. Progresses in genetics have permitted an improvement of management, screening and treatment. Surgery is the only successful treatment for MTC, as there is no effective adjuvant therapy for residual disease. A total thyroidectomy and vigilant management and surveillance of the neck are recommended. Interdisciplinary management including surgeons, endocrinologists, pathologists, radiotherapists, radiologists, and oncologists should be considered. PMID- 16472418 TI - Rectal bleeding by Dieulafoy-like lesion: successful endoscopic treatment. AB - A 81-year old woman affected by chronic renal failure, non insulin-dependent diabetes mellitus (NIDM) and hypertension, had an severe anemia massive hematochezia. The colonoscopy could not localize the bleeding site except some blood spots in the rectum. The patient was readmitted after 1 month with hypovolemic shock by massive hematochezia and required several blood transfusions. The endoscopic examination showed an important arterial bleeding treated successfully with epinephrine and bipolar elettro-coagulation (BICAP). We suggested that the patient presented a Dieulafoy-like lesion; this is an uncommon gastrointestinal cause of bleeding due to a defect of a submucosal artery without evidence of atherosclerosis or vasculitis. Both chronic renal failure and age could be considered as predisponent factors in this patient. Hematochezia is the most important sign and is often complicated by haemorrhagic shock. The diagnosis was delayed due to the difficulty in localizing the bleeding site; moreover, the patient needed several blood transfusions. The arteriographic diagnosis associated to endoscopic treatment by epinephrine and BICAP enabled a successful therapy. PMID- 16472419 TI - [Splenic marginal zone lymphoma: case report and review of the literature]. AB - Splenic marginal zone lymphomas are rare tumors which take origin from the B cells. More common in the elderly, often asymptomatic, they can present with abdominal pain, splenomegaly and cytopenia and have an indolent clinical course. We describe a case of a women 79 years old who presented with abdominal pain, fever and splenomegaly. Computed tomography demonstrated splenomegaly with an area of low density in the spleen. Only by laparotomy and splenectomy the correct diagnosis was possible. Because of the indolent course of this kind of lymphomas, splenectomy is the main treatment for patients with abdominal pain, splenomegaly and cytopenia. If there is no pain and no cytopenia, the treatment can be only wait and see. Only in case of progression of disease chemotherapy can be employed. PMID- 16472420 TI - [The capillary hemangioma of the stomach: an unusual cause of haemorrhage. Case report]. AB - Gastric haemorrhages are common clinical emergencies which often directly involve the surgeon in diagnosis and treatment; among these, rare vascular neoplasms deserve particular attention. The Authors report a rare case of benign vascular tumour of the stomach, a capillary angioma. After a literature review they highlight the importance of specific exams to reveal these small neoformations which, generally, show up clinically with acute bleeding, which may be severe, and which may not be discovered if the clinical evaluation is not very detailed. The surgical excision supported by a frozen sections is the most effective treatment. PMID- 16472421 TI - Eosinophilic cystitis associated with urethral stricture disease from pelvic trauma. Case report and literature review. AB - We report a case of eosinophilic cystitis (EC) in a 65-year-old man with urethral stricture disease from blunt pelvic traumatic event. EC is a rare condition characterized by eosinophilic infiltration of the bladder wall, that usually presents with irritative voiding symptoms, suprapubic pain and hematuria. Etiology is still not clear although a review of the literature suggests that pathogenetic mechanisms probably engage an altered immune response in the bladder, with the inflammatory reaction caused by factors such as exogenous allergens and previous bladder injury or surgery to the bladder or the prostate. The diagnosis of EC has to be confirmed by biopsy, since in some cases it may manifest as other inflammatory and malignant bladder disorders. A conservative medical management is indicated initially, since this disease may be self limited, with a benign course especially in children and young patients. In adults EC is more often a chronic recurrent condition that requires close follow up since it may lead to serious progressive bladder and/or upper urinary tract disease. More invasive therapies (including transurethral resection, partial or total cystectomy) may also be required when conservative therapy fails. PMID- 16472422 TI - [Results of surgical treatment of varicocele in male infertility]. AB - To evaluate the results of surgical treatment of varicocele on infertile men, especially regarding sperm count, 245 patients, surgically treated from 1993 to 2003, were evaluated. Patients underwent to ligature and section of the pampiniform plexus, through the subinguinal approach and local anaesthesia. At the follow-up (3-6-12 months) an improvement of sperm count was relieved in 79.5% of patients and the incidence of complications and relapses was of 3.7% and 1.2%, respectively. The Authors stress the efficacy of surgical treatment of varicocele in male infertility and hold the subinguinal approach as an effective treatment, minimally invasive and low cost. PMID- 16472423 TI - [Non recurrent laryngeal nerve. Personal experience]. AB - PURPOSE: Damage to the recurrent laryngeal nerve (RLN) during thyroid or parathyroid surgery is the most common iatrogenic cause of vocal cord paralysis. Identification of the RLNs and meticulous surgical technique can significantly decrease the incidence of this complication. Nonrecurrent RLNs (NRRLNs) are exceedingly rare. Surgeons need to be aware of their position to avoid injuries. PATIENT AND METHODS: A retrospective review of 263 right RLN exposures (and 251 left RNL) over a 5-year period was performed. RESULTS: Two NRRLNs were encountered, for an incidence of 0.39% (0.76% only for right dissection), without anatomic anomalies on the left side. The nerve anomaly was never preoperatively diagnosed. CONCLUSION: NRRLNs are rare and is associated with a right subclavian artery arising from distal aortic arch. Awareness of their existence and correct surgical technique will prevent the surgeon from accidentally lesion of NRRLN one if it is encountered during thyroid or parathyroid surgery. PMID- 16472424 TI - [Mininvasive treatment of strangulated paraesophageal hiatal hernia in emergency. Case report]. AB - The Authors report a case of strangulated paraesophageal hiatal hernia occurred in a elderly woman and treated with laparoscopic approach. After review of the literature regard on this uncommon pathology that present about 5% of the hiatal hernias, they emphasize that the laparoscopic approach is appropriated even in emergency and comprises complete reduction of the stomach in abdomen, control of suitable position of the distal esophagus and cardias and making of effective hiatus-plasty. PMID- 16472425 TI - [Longo hemorrhoidopexy vs Milligan-Morgan hemorrhoidectomy: perspective analysis]. AB - The authors analyzed their experience from January 2001 to February 2005 on 117 patients treated with Longo hemorrhoidopexy (46 cases) and Milligan-Morgan hemorroidectomy (71 cases). All the patients were observed after a week and one month after surgical procedure; at 6 months the Authors controlled 70 patients treated with Milligan-Morgan and 33 treated with Longo technique. The pain after 24 hours was the same in two groups but after a week a significative difference between two groups (p<0.05) was registered with a better quality of life for hemorrhoidopexy group. At 6 months pain during defecation was present in two cases of Longo group and in 6 cases of Milligan-Morgan group. In author's experience the Longo technique is a safe treatment with lower postsurgical pain and lower complications. PMID- 16472426 TI - [Endovascular laser ablation of the greater saphenous vein for varicose veins: our initial experience]. AB - Laser treatment of primary varicose veins of the legs is a new mini-invasive technique which represent an alternative to the safenectomy. Endovascular laser treatment is based on the employ of laser to destroying the vascular wall and inducing fibrosis. This technique is not without complications: burns, paraesthesias, haematomas, but most of all disappear in few days. Encouraged by the promising results reported in literature, we have performed 18 laser ablation of greater saphenous vein since 2003 till today. Our patients had a good post operative course and a follow up without troubles (3-17 months). We think that laser treatment is effective in the treatment of the primary varicose veins of the legs. It requests attention and experience in dosing the laser energy for minimizing the complications. Today there isn't long term follow up in literature. PMID- 16472427 TI - [Breast-conserving surgery in breast cancer]. AB - Breast-conserving surgery is the treatment of choice for the breast cancer T < 3 cm. The local recurrence is a problem of diagnosis and consequent treatment. We enrolled, from 1987 to 2004, 1504 breast cancer. In 803 (53.4%) tumor with T < 3 cm we performed conserving surgery. The sentinel lymph node (SN) technique induce to limit the axillary dissection in patients T1a-b to the SN only if non metastatic and located to the first level, the dissection of the 1st level of the axilla in patients T1c-T2 < 3 cm and SN negative, complete axillary dissection in patients with metastatic SN or located to the 2nd level. Our percentage of local recurrence in the follow-up was 3.5% at 5 years and 6% at 10 years. PMID- 16472430 TI - [Hypothermia--a promising supplement to the treatment of cardiac arrest]. PMID- 16472431 TI - [Registries and registration of clinical trials--secondary publication]. PMID- 16472432 TI - [Therapeutic hypothermia after cardiac arrest--status]. AB - Induced hypothermia in comatose patients resuscitated from cardiac arrest improves survival and neurological outcome when the initial rhythm is ventricular fibrillation. Results from 26 patients treated with early prehospital cooling, continued in hospital with cold fluids and body surface cooling, are presented. Sixteen patients had no or only minor neurological defects at discharge. Thirteen of 17 patients treated for ventricular fibrillation had a good cerebral outcome at discharge. Two patients had minor complications resulting from the treatment. PMID- 16472433 TI - [Non-classical human leukocyte antigen (HLA) tissue types--from implantation to transplantation]. AB - The classical and extremely polymorphic human leukocyte antigens (HLA) classes Ia and II have been studied in great detail and have significant importance in organ transplantation, autoimmune diseases and presentation of antigen peptides. However, in the human major histocompatibility complex (MHC), other functional HLA genes have been detected, the so-called non-classical HLA class Ib genes: HLA E, -G and -F. They resemble the HLA class Ia antigens in many ways, but several major differences have been described. They are almost monomorphic and generally have a restricted pattern of expression. One function of HLA-E is to inhibit and modulate natural killer (NK) activity. HLA-G seems to be important in the modulation of the maternal immune system during pregnancy and thereby the maternal acceptance of the semi-allogenic foetus. HLA-G can already be detected in at least some blastocysts and probably has a role in implantation. A very strong expression of HLA-G is observed in the invasive trophoblast cells in the placenta. HLA-G may be involved in certain complications of pregnancy and the genetic predisposition to these. Finally, HLA-G expression has been associated with a reduced risk of rejection episodes in heart and kidney/liver transplants. PMID- 16472434 TI - [Ethical aspects of transcultural research]. AB - An increasing number of patients in contact with the Danish health services have a non-Danish background. What are the ethical questions and dilemmas doctors face with this patient population in relation to their participation in medical research? This paper discusses the core medical ethical principles, including autonomy, beneficence, non-maleficence and justice in relation to the participation in research of members of various ethnic groups. PMID- 16472435 TI - [High-density lipoprotein: a new target in the battle against atherothrombotic disease]. AB - A low level of high-density lipoprotein (HDL) cholesterol is an important cardiovascular risk factor. The protective effects of HDL have been attributed to its promotion of reverse cholesterol transport, antioxidant activity and anti inflammatory actions on endothelial cells. Several treatment strategies aimed at exploiting the favourable effects of HDL are currently undergoing clinical trials and hold considerable promise as a novel approach to atheroprotection. PMID- 16472436 TI - [The influence of six alcoholic beverages on ethanol concentration in the blood and breath]. AB - INTRODUCTION: We investigated the area under the curve (AUC) as a measure of the bioavailability of ethanol in healthy volunteers drinking six alcoholic beverages of different types. Furthermore, we investigated the correlation between the ethanol concentration in the blood and the breath test. MATERIALS AND METHODS: Twelve healthy volunteers (seven females, five males) consumed six drinks of different types in a crossover design after a six-hour fast. The men ingested 36 g of ethanol and the women 24 g. Venous blood was obtained for determination of serum ethanol and glucose concentration at 0, 30, 60, 90, 120 and 180 minutes postdosing, and at the same time a breath alcohol test was done using an alcoholometer. RESULTS: The AUC of ethanol differed significantly between pure ethanol and the three beverages red wine, sparkling wine and Smirnoff Ice (p < 0.01). The glucose and insulin concentrations increased by factors of 2 and 4, respectively, 60 minutes after drinking beer or Smirnoff Ice, respectively (p < 0.05). We found a high correlation between the ethanol concentration in the blood and the breath test, r 2 = 0.77, r = 0.87 (p < 0.005). Using the blood alcohol concentration as a "gold standard", we found that the risk of obtaining a false- positive breath test was 1% at the legal limit for driving in Denmark (0.5%). Conversely, 59% of the participants with a blood alcohol level > or = 0.5% showed a negative (false negative). DISCUSSION: This investigation shows that the type of alcoholic beverage consumed determines the amount of alcohol absorbed. Furthermore, the different drinks caused different changes in the glucose and insulin concentrations, which might be important in connection with alcohol induced disturbances in carbohydrate metabolism (e.g., hypo- and hyperglycaemia). Our data indicate that the alcoholometer breath test was an acceptable screening method to estimate the blood alcohol level and to measure the amount of ethanol ingested. However, for evidental purposes during prosecution of drunk drivers, more sophisticated breath test instruments are desirable. PMID- 16472437 TI - [Prostate-specific antigen in the diagnosis of prostate cancer in Denmark]. AB - INTRODUCTION: The incidence of prostate cancer is increasing. Potentially curative treatment can be attempted in clinically localised disease, emphasising the role of early diagnosis. Prostate-specific antigen (PSA) is used as a biological marker for prostate cancer, but its value as a diagnostic test is associated with some degree of uncertainty. In 1999 the Danish Uro-Oncological Committee recommended measurement of the ratio of free to total PSA (f/tPSA) as a complementary analysis to total PSA (tPSA) in patients in whom the tPSA level is below 10 ng/mL. To assess whether the recommendation of the Danish Uro Oncological Committee has been implemented in daily clinical practice, we carried out a survey of a pre-selected sample of hospital laboratories. MATERIALS AND METHODS: Fifteen laboratories in Denmark, all associated with a university hospital, were surveyed in April 2004. Questions focused on current laboratory practice with regard to tPSA and f/t PSA. RESULTS: Within or outside their own facilities, the hospital laboratories all had tPSA analysed, and 64 percent had f/t PSA analysed. All the laboratories used the same critical value for tPSA (<4 ng/mL), while there was considerable variation in the critical value applied for f/tPSA. Forty-four percent of laboratories partially complied with the recommendation of the Danish Uro-Oncological Committee to analyse f/t PSA in addition to tPSA in patients in whom the total PSA level is below 10 ng/mL. DISCUSSION: Despite the recommendation of the Danish Uro-Oncological Committee, measurement of f/tPSA is not fully implemented in laboratories associated with university hospitals in Denmark. There is considerable practice variation in the analysis of f/t PSA as well. PMID- 16472438 TI - [Radio frequency of atrial fibrillation: results from 102 consecutive patients- secondary publication]. AB - Segmental pulmonary vein isolation (SPVI) and circumferential pulmonary vein isolation (CPVI) are the two main approaches to radiofrequency ablation in patients with atrial fibrillation (AF). The aim of this retrospective study was to investigate the safety level and patient outcome of both approaches. The overall freedom from symptoms was 73% and 25% after CPVI and SPVI, respectively. Our study concludes that CPVI is an effective treatment of highly symptomatic and drug-resistant AF and when used in laboratories by experienced electrophysiologists has a promising and reproducible outcome ratio and safety level. PMID- 16472439 TI - [Long-term survival rate of patients transfused with leucocyte-depleted and buffy coat-poor blood during colorectal surgery--secondary publication. An eight-year post-trial follow-up study]. AB - This study investigated the long-term survival rate of 589 patients enrolled in a trial in 1992-1995 who underwent colorectal surgery. The patients were randomised to receive leucocyte-depleted or buffy-coat-poor blood when transfusion was indicated. Significantly more of the non-transfused patients (59%) were alive seven years later compared to patients transfused with leucocyte-depleted blood (41%) and to patients transfused with buffy-coat-poor blood (45%). PMID- 16472440 TI - [Treatment of severe sarcoidosis using TNF-alpha-inhibitor (infliximab)]. AB - Some patients with severe sarcoidosis are resistant to conventional treatment with corticosteroids and cytostatics. We describe a 29-year-old man with histologically verified sarcoidosis and persistent, pronounced enlargement of the peripheral lymph nodes, resistant to treatment with prednisolone and methotrexate. Following treatment with TNF-alpha-inhibitor (infliximab 3 mg/kg body weight), the lymph nodes regressed to normal size and the patient improved clinically. The patient had a total of four infliximab infusions, and the effect has persisted for more than two years. PMID- 16472441 TI - [Serious bleeding in systemic lupus erythematosus complicated by lupus anticoagulant-hypoprothrombinaemia syndrome]. AB - Severe hemorrhagic diathesis due to lupus anticoagulant complicated by hypoprothrombinaemia resulting from prothrombin autoantibodies is a rare disorder and is often associated with systemic lupus erythematosus (SLE). We report a case in which a 15-year-old girl with SLE developed marked haemorrhagic manifestations due to menorrhagia and nosebleeds. The acute bleeding episode was treated with SAGM, tranexamic acid and recombinant factor VIIa. Lupus anticoagulant, cardiolipin antibodies and antiprothrombin antibodies were successfully depressed within weeks after corticosteroid therapy was begun. PMID- 16472442 TI - [Extrapancreatic tumour-induced hypoglycaemia]. AB - A 53-year-old non-diabetic man was admitted with hypoglycaemia, neuroglycopenic symptoms and acromegaloid facial swelling. Serum insulin concentration was suppressed, but the free concentration of insulin-like growth factor (IGF-II) was markedly elevated. CT scan demonstrated a large tumour in the liver. The histology showed a benign, solid, fibrous tumour. A hemihepatectomy was performed, and a 3.6 kg tumour was removed. Postoperatively, the blood glucose concentration and the concentration of free IGF-II returned to normal and the acromegaloid facial features disappeared. PMID- 16472447 TI - [The department of science undertakes a big responsibility]. PMID- 16472448 TI - [Physicians' advice on health--religion or science?]. PMID- 16472453 TI - [Emergency medicine specialty a possibility for quality improvement]. PMID- 16472450 TI - [The special feature issue on obesity]. PMID- 16472471 TI - The JIM interview. AB - Dr. Antonio Scarpa is director of the Center for Scientific Review (CSR) at the National Institutes of Health (NIH). He joined the CSR in July 2005 after 19 years at Case Western Reserve University in Cleveland, Ohio. Scarpa earned his medical degree and PhD in general pathology from the University of Padua School of Medicine and continued his training at the Weizmann Institute of Science in Israel, the University of Utrecht in the Netherlands, the University of Bristol in England, and the University of Pennsylvania in Philadelphia. He was appointed to the faculty of the University of Pennsylvania and continued his research there for 17 years. In 1986, Scarpa joined Case Western, where he was the David and Inez Myers Professor and chair of the Department of Physiology. PMID- 16472473 TI - National Institutes of Health embarks on a 3-Year, 100 million dollars pilot exploration of cancer genomics. PMID- 16472474 TI - Thiazolidinediones and progression of renal disease in patients with diabetes. AB - OBJECTIVE: Thiazolidinediones (TZDs) are used in the treatment of type 2 diabetes mellitus (T2DM) and appear to have beneficial effects on markers of cardiovascular or renal risk that are independent of glycemic control. We examined the effects of TZDs on renal survival in a predominantly black population with T2DM. METHODS: We performed a retrospective case-control study in patients with T2DM seen in our nephrology clinic in 2001 to 2002. Cases had T2DM and were on a TZD at presentation or for >or= 6 months over follow-up. Controls were matched for sex, age, duration of T2DM, and initial creatinine. Reaching end stage renal disease (ESRD) was the primary end point. RESULTS: From 387 records, 43 cases (34 blacks, 31 females) and 106 controls (96 blacks, 83 females) were identified. The baseline characteristics were similar for both groups. Both groups had moderate renal disease (estimated glomerular filtration rate approximately 40-45 mL/min). Cases had lower systolic blood pressure over follow up (p=.02), but there was no difference in glycemic control or use of insulin. Renal survival was better among cases (age- and gender-adjusted odds ratio for reaching ESRD 0.17 [95% confidence interval 0.03-0.8]; p=.03). When adjusted for systolic blood pressure over follow-up, the tendency for improved renal survival in cases remained but was no longer significant. CONCLUSION: We conclude that TZDs may protect against the progression of renal disease in T2DM. Prospective studies are required to determine the effects of TZDs on renal survival in T2DM. PMID- 16472475 TI - Influence of the dialysis membrane on markers of tissue ischemia. AB - BACKGROUND: Hemodialysis (HD) is often accompanied by adverse effects, such as tissue ischemia. We have already observed an increase in plasma adenosine (ADO) levels during HD sessions, which may be the result of tissue ischemia. Here we evaluate the influence of the dialysis membrane on two sensitive and early markers of ischemia: ADO and ischemia-modified albumin (IMA). METHODS: We included in the study 50 patients with end-stage renal failure, 39 hemodialyzed (mean age 61+/-24 years; 24 male; membranes: 23 synthetic and 16 cellulose based) and 11 undialyzed (mean age 55+/-12 years; 6 male), and 10 healthy subjects (mean age 47+/-11 years; 4 male). We compensated for hemoconcentration during HD by measuring either the IMA to albumin (Alb) or the ADO to Alb ratio. RESULTS: Under basal conditions, the IMA to Alb ratio was not significantly different in patients and controls and HD did not significantly modify this ratio. Conversely, the ADO to Alb ratio (mean+/-SD in micromol/g) was higher in patients before HD compared with either undialyzed patients or controls (before HD: 0.077+/-0.02; undialyzed patients: 0.026+/-0.11; controls: 0.022+/-0.01). During HD, there was a significant increase in the ADO to Alb ratio (before HD: 0.077+/-0.02; after HD: 0.09+/-0.029; p<.01). We found no significant difference in the IMA to Alb or ADO to Alb ratio using either synthetic or cellulose-based membranes. CONCLUSIONS: We concluded that ADO is a more sensitive marker of ischemia than IMA and that, under our HD conditions, the ischemia caused by HD was very weak, independent of the dialysis membrane. PMID- 16472476 TI - Exercise, caloric restriction, and systemic oxidative stress. AB - BACKGROUND: In humans, the main sources of reactive oxygen species (ROS), the molecules causing oxidative stress, are mitochondrial superoxide ions and neutrophil-derived oxidative radicals. Circulating antioxidants contribute to the protection against oxidative stress. Although the formation of ROS and secretion of antioxidants are independently regulated by exercise and diet, little is known about their combined effect. We hypothesized that relatively brief, intense exercise training may reduce systemic oxidation via an intrinsic mechanism, independent of changes in circulating antioxidants and of neutrophil-derived enzymes (as may be caused by concomitant caloric restriction). METHODS: Nineteen volunteers exercised for 7 days, 3 hours/day at 75% of oxygen uptake. Caloric intake was either 110% of caloric expenditure (high calorie, n=10) or 75% of caloric expenditure (low calorie, n=9). Blood samples for F2-isoprostanes, catalase, myeloperoxidase (MPO), interleukin-x (IL-x), white blood cells (WBCs), and other metabolic variables were taken at baseline, at the end of training, and 1 week after completion of the study. RESULTS: Serum F2-isoprostanes (microg/mL), markers of lipid peroxidation, were similarly reduced after 7 days of exercise in the high-calorie (from 35+/-4 to 27+/-2) and low-calorie (from 35+/-3 to 24+/-2) groups. Similar reductions were observed in IL-x concentrations. Conversely, no change was observed in circulating concentrations of the antioxidant catalase. Whereas total WBCs and neutrophil counts were significantly reduced in the low calorie group only, no difference in neutrophil-derived MPO was measured between groups. CONCLUSION: A significant reduction in systemic oxidation may occur relatively early during intense exercise training in healthy young men, independent of caloric intake. The potential contribution to these effects of circulating antioxidants and neutrophil-derived oxidative enzymes will require further investigation. PMID- 16472477 TI - Differences in triage thresholds for patients presenting with possible acute coronary syndromes: more than meets the eye. AB - BACKGROUND: Many studies have shown differences in cardiac care by racial/ethnic groups without accounting for institutional factors at the location of care. OBJECTIVE: Exploratory analysis of the effect of hospital funding status (public vs private) on emergency department (ED) triage decision making for patients with symptoms suggestive of acute coronary syndromes (ACSs) and on the likelihood of ED discharge for patients with confirmed ACS. STUDY DESIGN AND SETTING: Secondary analysis of data from a randomized controlled trial of 10,659 ED patients with possible ACS in five urban academic public and five private hospitals. The main outcome measures were the sensitivity and specificity of hospital admission for the presence of ACS at public and private hospitals and the adjusted odds of a patient with ACS not being hospitalized at public versus private hospitals. RESULTS: Of 10,659 ED patients, 1,856 had confirmed ACS. For patients with suspected ACS, triage decisions at private hospitals were considerably more sensitive (99 vs 96%; p<.001) but less specific (30 vs 48%; p<.001) than at public hospitals. The difference between hospital types persisted after adjustment for multiple patient-level and hospital-level characteristics. CONCLUSION: Significant differences in triage for patients with suspected ACS exist between public and private hospital EDs, even after adjustment for multiple patient demographic, clinical, and institutional factors. Further studies are needed to clarify the causes of the differences. PMID- 16472478 TI - Molecular mechanisms linking sodium to hypertension: report of a symposium. AB - There is abundant clinical and epidemiologic data linking excess body sodium with hypertension. The mechanism(s) at the molecular level to explain this relationship are unknown. Recent studies by multiple investigators, have identified several ion transport mechanisms in the vascular wall that interact to control vascular tone and contractility. These new data include 1) biochemical, pharmacologic, and molecule structural studies, 2) experiments in transgenic and knockout mice, and 3) results in clinical hypertension. The overall results provide compelling evidence for the concept that salt-dependent hypertension involves the secretion of endogenous ouabain (EO), an adrenal steroid synthesized with the same initial steps as aldosterone and secreted by the zona glomerulosa. Circulating EO inhibits arterial smooth muscle Na+ pumps with alpha 2 subunits. These are functionally coupled to the type 1 Na/Ca exchanger (NCX1). Thus when a2 Na pumps are inhibited in arterial smooth muscle, the resulting subplasma membrane increase in Na+ concentration triggers, via NCX1 Ca2+ entry, a rise in cytosolic Ca2+ concentration and increased myogenic tone and contractility. The ultimate result is a rise in peripheral vascular resistance-the hemodynamic hallmark of hypertension. The elucidation of this pathway has facilitated the development of pharmacologic agents that have therapeutic potential for hypertension and other cardiovascular diseases. These include agents that compete with EO for binding to the Na+ pump and inhibitors of NCX1. PMID- 16472480 TI - [Preventable adverse drug events in hospitalized patients]. AB - BACKGROUND AND OBJECTIVE: To determine the incidence of adverse drug events (ADE) in hospitalized patients, identify those that were potentially preventable, and asses the drug classes involved, the clinical symptoms and the type of medication errors that led to the preventable ADE. PATIENTS AND METHOD: An observational study of ADE prevalence in hospitalized patients in internal medicine, pneumology, gastroenterology, nephrology and neurology wards, over a six-month period, at a tertiary university hospital. ADE were prospectively detected through physician and nurses reporting fostered by daily visits of a clinical research and retrospectively through review of medical records using event codes as defined by the IDC-9-CM system. RESULTS: In a total of 2,643 hospitalized patients, 191 (7.2%) ADE were detected. Of these, 38 cases (19.9%) were classified as preventable, of which 21.1% were mild; 60.5% moderate and 18.4% serious or life-threatening. Preventable ADE were frequently associated with anti infective drugs (22.9%), diuretics (18.8%) and digoxin (16.7%). Inadequate therapy monitoring (28.3%), excessive dosage (21.7%), selection of an inappropriate drug according to patient characteristics and/or to diagnosis (15.0%), lack of prescription of a necessary drug (15.0%) and drug-drug interactions (11.7%) were the most common identified type of errors leading to preventable ADE. CONCLUSIONS: 1.4% of hospitalized patients in medical wards experienced potentially preventable ADE. Healthcare professionals and administrators must be made aware of the scope of this problem so that they will implement effective safety practices directed to reduce the incidence of medication errors, particularly prescription and monitoring errors. PMID- 16472481 TI - [Direct discharge from triage in emergency departments: assessment, risks and patient satisfaction]. AB - BACKGROUND AND OBJECTIVE: After triage assessment, some hospitals refer emergency department (ED) patients with minor chief complaints to off-site clinics. The potential for 2 different referral models introduced in 2 urban hospitals was assessed, as well as the suitability of this measure. PATIENTS AND METHOD: After triage assessment, patients with minor complaints were identified. ED from Hospital Clinic of Barcelona (HCB) proposed that these patients should be referred to a hospital affiliated off-site clinic, whereas ED from Hospital Mutua de Terrassa (HMT) tried to refer such patients to their primary care setting. Within a year, we assessed on both ED the following items: number of arrivals, percentage of proposed referrals (PR), percentage of accepted referrals (AR), percentage of carried-out referrals (CR: patients who, once discharged, really attended the other setting), and percentage of returned referrals (RR) to the ED, as well as return reasons, and the percentage of returned patients finally admitted. The degree of patient satisfaction was evaluated by means of a telephone survey. RESULTS: From both ED, 44,764 arrivals and 7,297 (16.3%) PR were registered. The percentage of AR and CR was 94.3% and 75.3%, respectively. The percentage of PR from HMT was higher (18.7% vs 13.1%; p < 0.001), yet HCB obtained a greater percentage of AR (98.9% vs 92%, p < 0.001) and CR (93.7% vs 65%; p < 0.001). The percentage of RR from both ED was 1.5% (2.8% vs 0.4%; p < 0.001). Among returned patients, 12 of them (0.17% respect to AR) were finally admitted. Only 41% of patients who were found to be eligible for direct discharge would have agreed with being referred to another clinical setting, but after the experience, up to 93% of them said they would go through it again. HMT gave referral information to patients faster than HCB (p < 0.05), but HCB got a better model evaluation (p < 0.01), a greater medical complaint solution (p < 0.05), and generated a smaller number of subsequent consultations (p < 0.05). CONCLUSIONS: After a rapid triage assessment, a percentage of patients arriving on EDs may be directly and safely discharged and referred to other clinical settings. This is achieved by both proposed models without additional risks for patients. However, patients feel more satisfied with an hospital-affiliated off-site clinic instead of their primary care setting. PMID- 16472482 TI - [Reemergence of infectious syphilis among homosexual men and HIV coinfection in Barcelona, 2002-2003]. AB - BACKGROUND AND OBJECTIVES: An increase in syphilis infections since the mid 1990s has been documented, especially in homosexual men, in different European and North American cities. We intended to describe the characteristics of newly diagnosed cases of syphilis at the Sexually Transmitted Infections Unit of Barcelona in 2002 and 2003. PATIENTS AND METHOD: Descriptive analysis of cases with infectious syphilis and multivariate analysis of factors associated with HIV coinfection. RESULTS: 102 cases were diagnosed with infectious syphilis, 98 males (88 homosexual men). HIV coinfection was present in 34% of cases. Predictive factors of HIV coinfection were age > 30 years (p = 0.003) and having a HIV positive partner (p = 0.044). Clinically, there were no differences between cases coinfected or not with HIV. CONCLUSIONS: There has been a recent increase of syphilis in Barcelona, especially among some core groups of homosexual men with high rates of HIV coinfection. PMID- 16472483 TI - [What we may we learn. Drug-related adverse events?]. PMID- 16472484 TI - [Cardiovascular disease and hypertensive cardiomyopathy. Beyond heart]. PMID- 16472485 TI - [Early screening of intimate partner violence. An evidence-based intervention?]. PMID- 16472486 TI - [Home blood pressure measurement]. AB - Measurement of blood pressure (BP) by the own patient at home has become a popular procedure. Both a great offer of simple electronic devices and publication of multiple studies supporting its usefulness for the diagnosis and control of hypertension have contributed to this fact. In fact, this technique has various advantages over the conventional clinical measurement such as a higher reproducibility, better representation of the usual BP profile of the patient, greater correlation with target organs damage and greater morbimortality predictive value. Besides, there is a potential benefit on the control of patients under antihypertensives and, maybe, a reduction in health expenses at long-term follow-up. Currently, the main hypertension societies recommend the home measurement of BP in some clinical situations. However, it cannot be considered an alternative to the clinical measurement but rather a complementary technique in the evaluation of the hypertensive patient. Here we review the indications and limitations of this procedure in clinical practice. PMID- 16472487 TI - [Acute behavior disorder and hypopotassemia in a 49-year-old male]. PMID- 16472488 TI - [Abnormal glycemic metabolism in essential hypertension. Role of the oral glucose tolerance test]. PMID- 16472489 TI - [Abnormal glycemic metabolism in essential hypertension. Role of the oral glucose tolerance test]. PMID- 16472491 TI - [When the cat jumps out of the boat]. PMID- 16472493 TI - [Talking about smoking: a proposal of improvement of the concept "packyear"]. PMID- 16472494 TI - [Influence of antecedent of hypertension in patients with acute coronary syndrome without ST elevation]. AB - BACKGROUND AND OBJECTIVE: There are few studies evaluating the effect of a previous history of hypertension on long term prognosis after an acute coronary syndrome, using the new definitions and incorporating new risk markers in the analysis. The aim of our study was to determinate if hypertensive patients differ from non-hypertensives in the epidemiological profile, clinical presentation, treatment prescribed at discharge and prognosis after admission with non ST segment elevation acute coronary syndrome. PATIENTS AND METHOD: A total of 1,029 consecutive patients admitted with high suspicion of non ST segment elevation acute coronary syndrome were evaluated. Prognostic variables were determined during admission (epidemiological and biochemical), as it was the discharge treatment. The primary endpoint was defined as all cause mortality at one year follow up. RESULTS: 65.8% (n = 677) of patients had hypertension. Hypertensive patients displayed a worst epidemiological and biochemical profile, and different discharge treatment. There were 139 (13.5%) deaths at one year follow up. The all cause mortality for non-hypertensive patients was 12.5% and for hypertensives 14.6% (p = NS). In the multivariate analysis (Cox regression) there were no differences in mortality between these groups. CONCLUSIONS: A previous history of hypertension is an important factor to explain differences in the presence of other risk factors or the treatment, but is not a mortality predictor. PMID- 16472495 TI - [Utilization of patient isolation in non critical units from a university hospital]. AB - BACKGROUND AND OBJECTIVE: Correct utilization of source isolation (SI) in hospitals is important to take advantage of hospital resources. The objective of this work was to evaluate the utilization of SI considering the appropriateness of isolation length-stay. PATIENTS AND METHOD: During a period of 19 months a prospective observational study was carried out among non-critical inpatients who underwent SI in a university hospital. The information was obtained from Admission Unit data, daily ward rounds and review of case history records. RESULTS: A total of 239 SIs were identified, summarizing 2,589 days in isolation (median: 8 days, range: 1-56 days). These data supposed a cumulative incidence of 6.74 isolations by 1,000 admissions, and an incidence density of 1.08 isolations by 1,000 inpatient-day. 36.8% of SIs were considered incorrectly used, meaning a total of 703 inappropriate stays in isolation (27.1% of all stays carried out in SI). Surgical wound infections prompted inappropriate isolations with a greater frequency (45.2%), mainly after hip and knee prosthesis implantation interventions (42.9% and 17.9%, respectively). CONCLUSIONS: It is necessary to develop quality criteria and indicators in order to implement quality improvement actions to optimize SI length-stay management. PMID- 16472496 TI - [Methicillin resistant Staphylococcus aureus infection postoperatory complications and prognosis of patients with lower extremity amputations]. AB - BACKGROUND AND OBJECTIVE: Methicillin-resistant Staphylococcus aureus (MRSA) is associated with an increasing morbimortality when compared with other microorganisms. The aim of this study was to examine the complications and prognosis of the presence of MRSA in vascular patients with amputation of lower limbs. PATIENTS AND METHOD: We included patients who had lower extremity amputation in our department in 2004 and displayed positive surgical wounds cultures. We compared patients with MRSA positive cultures with other microrganisms. We evaluated general characteristics, operative indications, surgical wounds microbiology, reamputations, morbimortality and mean time of stay in hospital. RESULTS: 117 patients (median age 73, 68% male) underwent lower extremity amputation. 82 of them had positive cultures and MRSA were isolated in 30% cases. Two two groups were comparable and no statistical differences were found in relation to reamputation rate, morbimortality and mean time of stay in hospital. CONCLUSION: Presence of MRSA does not represent an additional risk of reamputation or an increase of postoperative complications. Careful wound surveillance, through wound debridement and optimal administration of antibiotics must be applied to all patients, regardless of the bacterial flora. PMID- 16472497 TI - [Is resynchronization possible in Spain?]. PMID- 16472498 TI - [Therapeutic usefulness of sentinel lymph node biopsy in patients with melanoma]. PMID- 16472499 TI - [Celiac disease]. PMID- 16472500 TI - [Prevention and control of chemotherapy-induced nausea and vomiting]. AB - Nausea and vomiting are considered one of the most distressing side-effects of chemotherapy. Complete control of acute and delayed emesis improves quality of life and increases adherence to treatment. The frequency of nausea and vomiting depends primarily on the emetogenic potential of the chemotherapeutic agents used. With the standard antiemetic therapy (5HT-3 receptor antagonists in combination with dexamethasone) approximately 13% of patients receiving chemotherapy have vomiting in the acute phase and almost 50% in the delayed phase. A new group of antiemetic drugs, the neurokinin-1 receptor antagonists, in combination with standard therapy significantly improves emesis protection in the acute and in the delayed phase, although control of nausea is not so effective. Nowadays chemotherapy-induced emesis still occurs. Recent developments in antiemetic therapy and responsibility to achieve the best control of nausea and vomiting in patients receiving chemotherapy justified a review of this problem, which is frequently underestimated by physicians and nurses. PMID- 16472501 TI - [Medicolegal aspects of medical responsibility]. PMID- 16472502 TI - [Antiretroviral therapy in patients with primary HIV infection (corrected)]. PMID- 16472503 TI - [Antiretroviral treatment in patient with advanced immunosuppression]. PMID- 16472505 TI - [Trends in the consumption of tobacco, alcohol and cannabis among adolescents of Barcelona, Spain, between 1987 and 2004]. PMID- 16472518 TI - Ruminant alphaherpesviruses related to bovine herpesvirus 1. AB - Herpesviruses have mainly co-evolved with their hosts for millions of years. Consequently, different related host species may have been infected by various genetically related herpesviruses. Illustrating this concept, several ruminant alphaherpesviruses have been shown to form a cluster of viruses closely related to bovine herpesvirus 1 (BoHV-1): namely bovine herpesvirus 5, bubaline herpesvirus 1, caprine herpesvirus 1, cervid herpesviruses 1 and 2 and elk herpesvirus 1. These viruses share common antigenic properties and the serological relationships between them can be considered as a threat to BoHV-1 eradication programmes. BoHV-1 is a herpesvirus responsible for infectious bovine rhinotracheitis, which is a disease of major economic concern. In this article, the genetic properties of these ruminant alphaherpesviruses are reviewed on a comparative basis and the issue of interspecific recombination is assessed. The pathogenesis of these infections is described with emphasis on the host range and crossing of the host species barrier. Indeed, the non bovine ruminant species susceptible to these ruminant alphaherpesviruses may be potential BoHV-1 reservoirs. The differential diagnosis of these related infections is also discussed. In addition, available epidemiological data are used to assess the potential of cross-infection in ruminant populations. A better knowledge of these ruminant alphaherpesvirus infections is essential to successfully control infectious bovine rhinotracheitis. PMID- 16472519 TI - Genetic evolution of canine coronavirus and recent advances in prophylaxis. AB - Since the first identification of the virus in 1971, the disease caused by canine coronavirus (CCoV) has not been adequately investigated and the role that the virus plays in canine enteric illness has still not been well established. In the last decade, as a consequence of the relatively high mutation frequency of RNA positive stranded viruses, CCoV has evolved and a new genotype has been identified in the faeces of infected dogs. The several studies carried out by different researchers have focused upon the epidemiological relevance of these viruses and, considering the wide diffusion of CCoV infections among dog populations, the author underlines the need for further investigation on the biology of CCoV and on the pathogenetic role of their infections. PMID- 16472520 TI - Corynebacterium pseudotuberculosis: microbiology, biochemical properties, pathogenesis and molecular studies of virulence. AB - Corynebacterium pseudotuberculosis is the etiological agent of caseous lymphadenitis (CLA), a common disease in small ruminant populations throughout the world. Once established, this disease is difficult to eradicate because drug therapy is not effective and because the clinical detection of infected animals is of limited efficiency. We reviewed the microbiological, biochemical and taxonomic features of C. pseudotuberculosis, general aspects of infection, the main virulence determinants and currently available commercial vaccines. We also examined the current molecular strategies for the study of virulence in C. pseudotuberculosis, including the latest research on the identification of novel virulence factors and genes, which will help us to better understand the biology of this microorganism. This knowledge may also contribute to the development of improved CLA vaccines, including subunit and DNA-based types, as well as to improve the diagnosis, treatment and control of this disease. PMID- 16472521 TI - Characterization of cytokine expression in milk somatic cells during intramammary infections with Escherichia coli or Staphylococcus aureus by real-time PCR. AB - The expression of inflammatory cytokines, including interleukin (IL)-6, IL-8, IL 12, granulocyte macrophage-colony stimulating factor (GM-CSF), tumor necrosis factor (TNF)-alpha, and interferon (IFN)-gamma, by milk somatic cells was characterized by real-time polymerase chain reaction (PCR) in dairy cows experimentally challenged with either E. coli (n = 8) or S. aureus (n = 8). The mRNA abundance of a target gene was calibrated with that of a reference gene (beta-actin) and expressed as fold of induction over the control quarter at each time point. At no single time point did all eight quarters challenged with the same type of bacteria demonstrated increased expression of a target gene and there was large variation among animals at each given time. As a consequence, most tested comparisons were not statistically significant except the peak time points of IL-8 expression (75- and 29- fold in glands challenged with E. coli and S. aureus, respectively). However, the average fold induction of all targeted cytokines was increased in response to both bacterial challenges with the exception of IFN-gamma. The expression of IFN-gamma was only increased in milk somatic cells isolated from E. coli, but not S. aureus, challenged mammary glands. Moreover, upregulated expression of cytokine genes had higher magnitudes and/or faster responses in glands challenged with E. coli in comparison with those challenged with S. aureus. We propose that the compromised upregulation of inflammatory cytokines in S. aureus infected glands may, at least partially, contribute to the chronic course of infection caused by this pathogen. Further research on identifying factors responsible for the differentially expressed cytokine profiles may be fundamental to developing strategies that mitigate the outcome of bovine mastitis. PMID- 16472523 TI - Broad-range PCR-TTGE for the first-line detection of bacterial pathogen DNA in ticks. AB - Ticks are known or suspected vectors for a wide range of bacterial pathogens. One of the first steps for tick-borne risk assessment is the detection of these pathogens in their vectors. In the present study, a broad-range PCR amplification of the eubacterial gene encoding the 16S rRNA gene combined with Temporal Temperature Gradient gel Electrophoresis (TTGE) was evaluated as a method allowing the one-step detection of bacterial pathogen DNA in ticks. Firstly, DNA extracts from bacteria known to be tick-borne pathogens, i.e., Borrelia burgdorferi lato sensu, Anaplasma phagocytophilum, Spotted Fever Group (SFG) Rickettsia spp., were used to establish a TTGE pathogen DNA reference marker. Secondly, we used broad-range PCR-TTGE to detect the presence of DNA from these three pathogens in 55 DNA extracts from pools of 10 nymphal Ixodes ricinus ticks, which have been previously shown to carry DNA from at least one of those bacteria by specific PCR. Among the 20 B. burgdorferi specific-PCR samples, 15 (75%) were also found to be positive using PCR-TTGE. Sixteen of the seventeen (94%) Rickettsia spp. PCR-specific samples were positive using PCR-TTGE detection and all PCR-specific positive extracts (11/11, 100%) for A. phagocytophilum were also positive using PCR-TTGE. Moreover, we identified unexpected bacterial sequences that were not related to any of the three pathogens such as a sequence related to Spiroplasma sp. Thus, broad-range PCR-TTGE allowed the single step detection of DNA from up to 3 pathogens in the same co-infected samples as well as detection of DNA from unexpected bacteria. PMID- 16472522 TI - A serological study of exposure to arthropod-borne pathogens in dogs from northeastern Spain. AB - There is limited information regarding the prevalence of many vector borne pathogens in Europe and especially in Spanish dogs. We investigated 206 sick and 260 clinically healthy dogs from three different regions in northeastern Spain for antibodies to Rickettsia conorii (Rc), Ehrlichia canis (Ec), Anaplasma phagocytophilum (Ap), Bartonella henselae (Bh), Bartonella vinsonii subsp. berkhoffii (Bvb), Leishmania infantum (Li) and Borrelia burgdorferi (Bb) and for antigen of Dirofilaria immitis (Di). Total prevalences were the following: Rc (56.4%), Li (30%), Ec (16.7%), Bh (16.8%), Ap (11.5%), Bvb (1.07%), Di (0.6%) and Bb (0.6%). Seroprevalences for Rc, Ec, Ap, Bh, and Bvb and Bb and Di antigens were similar among the three different study sites. The Ec seroprevalence, as determined by Snap 3DX, was statistically lower in dogs from Mallorca (0%) than Tarragona (16%) and Barcelona (5%) (P < 0.0001). Detection of Rc antibodies was associated with seroreactivity to Ec and Ap antigens (P = 0.018 and P = 0.002, respectively). IFA Ec antibodies were associated with Ap seroreactivity (P < 0.0001). There was no association between the clinical status, sex, time of the year when samples were collected, life-style or exposure to fleas or ticks and a positive test result for Ec, Bh, Bvb, or Bb antibodies or Di antigens. Li seroreactivity was associated with illness and living outdoors (P < 0.0001, P = 0.029; respectively), Rc seroreactivity with the male gender (P = 0.028) and Ap seroreactivity with living outdoors (P = 0.045). This study indicates that exposure to Rc, Li, Ec or related Ehrlichia spp., Bh and Ap or a related spp., is common whereas Di, Bb and Bvb is uncommon among dogs from the Mediterranean basin. We also provide serological data that suggests the existence of a novel Ehrlichia species on Mallorca island. PMID- 16472524 TI - Mechanisms of chronic waterborne Zn toxicity in Daphnia magna. AB - In order to gain better insights in the integrated response of Daphnia magna following chronic zinc exposure, several physiological parameters were measured in a time-dependent manner. D. magna juveniles were exposed for 21 days to dissolved Zn concentrations up to 340 microg/L. Next to standard endpoints such as mortality, growth and reproduction the following sub-lethal endpoints were measured: filtration and ingestion rate, respiration rate, energy reserves, internal Zn and total Ca concentrations in the organisms. Organisms exposed to 80 microg/L generally performed better than the Zn deprived control organisms. The former were used to elucidate the effects of higher Zn concentrations on the endpoints mentioned above. After 1 week, only 7% of the organisms exposed to 340 microg/L survived. Body Zn contents of these organisms were 281 +/- 76 microg g dry weight and a 37% decrease of the Ca contents was observed. This suggests a competitive effect of Zn on Ca uptake. Filtration rate (-51%), individual weight (-58%) and energy reserves (-35%) also exhibited a decreasing trend as a function of increasing Zn exposure concentrations. During the second and third exposure week an overall repair process was observed. In the surviving organisms mortality and reproduction were only slightly affected. This can be explained by (over)compensation reactions at lower levels of biological organisation: Ca contents (+24%) and filtration rate (+90%) increased as a function of the exposure concentration while respiration rate decreased (-29%) resulting in energy reserves remaining constant as a function of Zn exposure. It is hypothesized that a disturbed Ca balance is probably the first cause for zinc toxicity effects in D. magna. PMID- 16472526 TI - Determination of subtalar joint axis location by restriction of talocrural joint motion. AB - The location of the subtalar joint axis is an important determinant of the mechanical function of the foot. The moments of muscle forces and of the ground reaction force about the subtalar joint are dependent upon the location of this joint axis. There is substantial variation in subtalar axis location across subjects, but current methods for determining its location are often invasive or involve expensive imaging protocols. A novel technique for location of the subtalar axis is presented in which the talocrural joint is passively immobilized so that motion of the tibia relative to the calcaneus can be used to estimate the subtalar axis. This paper presents results of cadaver testing in which accuracy of the technique was assessed by comparing helical axes computed from calcaneus tibia bone motions to axes computed from calcaneus-talus bone motions. Only small motions at the talocrural joint were observed, and good estimates of the subtalar axis (errors less than 15 degrees and 2mm) were achieved in four of six specimens. PMID- 16472525 TI - Training induced adaptations in characteristics of postural reflexes in elderly men. AB - The aging neuromuscular system is affected by structural and functional changes which lead to a general slowing down of neuromuscular performance and an increased risk of falling. The impact of heavy resistance (HR) training in the elderly on maximum voluntary contraction (MVC) and rate of force development (RFD) has been investigated in the past. However, the influence of sensorimotor (SENSO) training and HR training on the ability to compensate for gait perturbations has not yet been investigated in the elderly. Therefore, the purpose of the study was to examine the impact of HR and SENSO training in elderly men on unexpected treadmill perturbations. Functional reflex activity was recorded by means of surface EMG in 40 male subjects (>60 years) before and after 13 weeks of HR and SENSO training and in another 20 male subjects (>60 years), which served as a CONTROL-group. SENSO training resulted in a decrease in onset latency, an enhanced reflex activity in the prime mover as well as a decrease in maximal angular velocity of the ankle joint complex during the perturbation impulses. No significant changes were observed in the HR- and in the CONTROL group. The results clearly indicate that SENSO training has an impact on spinal motor control mechanisms in the elderly. Training induced improvements in perception and procession of afferent information could be a possible reason for the increase in reflex contraction. Due to these adaptive processes, SENSO training could be a well-suited method for fall preventive programs in elderly people. PMID- 16472527 TI - Simple gas chromatographic method for the stereodifferentiation of methyl nilate, a chiral alpha-methyl-beta-hydroxy ester. AB - The four stereoisomers of methyl nilate, methyl 3-hydroxy-2-methylbutanoate, have been separated and analyzed by the direct simultaneous achiral-chiral high resolution gas chromatography (HRGC) and also by the chiral high-resolution gas chromatography-time-of-flight mass spectrometry (HRGC-TOF-MS). The method involved the use of DB-5 and CP Chirasil-Dex CB columns. The elution sequences on the two columns were determined and are reported. The one-step GC method allows the identification by retention data and simultaneously to determine unequivocally the absolute configuration of nilic acid moieties contained in minor complex natural products without previous isolation, as part of a general strategy for the structure determination of the compounds. PMID- 16472528 TI - Liquid phase microextraction with in situ derivatization for measurement of bisphenol A in river water sample by gas chromatography-mass spectrometry. AB - A new method that involves liquid phase microextraction (LPME) with in situ derivatization and gas chromatography-mass spectrometry (GC-MS) is described for the determination of trace amounts of bisphenol A (BPA) in river water samples. The LPME conditions, such as the type of extraction solvent and the extraction time, are investigated. Then, the extract is directly injected into GC-MS. The detection limit and the quantification limit of BPA in river water sample are 2 and 10pgml(-1) (ppt), respectively. The calibration curve for BPA is linear with a correlation coefficient of >0.999 in the range of 10-10,000pgml(-1). The average recoveries of BPA in river water samples spiked with 100 and 1000pgml(-1) BPA are 104.1 (RSD: 8.9%) and 98.3 (RSD: 3.2%), respectively, with correction using the added surrogate standard, bisphenol A-(13)C(12). This simple, accurate, sensitive and selective analytical method may be applicable to the determination of trace amounts of BPA in liquid samples. PMID- 16472529 TI - Analysis by gas chromatography-mass spectrometry of the volatiles from the fruits of Ammodaucus leucotrichus subsp. leucotrichus and subsp. nanocarpus grown in North Africa and the Canary Islands, respectively. AB - The volatiles from the fruits of Ammodaucus leucotrichus subsp. leucotrichus and subsp. nanocarpus (two endemic species, the first from North Africa and the second from the Canary Islands, Spain) were studied by gas chromatography and gas chromatography-mass spectrometry. The major components of the volatiles of subsp. nanocarpus were found to be, beta-pinene (22.2-33.6%), bornyl angelate (20.6 21.8%) and camphor (8.3-11.7%) whereas in the fruits of subsp. leucotrichus, the main constituents were perillaldehyde (63.6%) and limonene (26.8%). We also suggest that subsp. nanocarpus should have the status of species and should be named Ammodaucus nanocarpus. PMID- 16472530 TI - Amperometric detector designs for capillary electrophoresis microchips. AB - Electrochemical (EC) detection is a sensitive and miniaturisable detection mode for capillary electrophoresis (CE) microchips. Detection cell design is very important in order to ensure electrical isolation from the high separation voltage. Amperometric detectors with different designs have been developed for coupling EC detection to CE-microchips. Different working electrode alignment: in channel or end-channel has been tested in conjunction with several materials: gold, platinum or carbon. The end-channel detector was based on a platinum or gold wire manually aligned at the exit of the separation channel. Thick- (screen printed carbon electrode) and thin-film (sputtered gold film) electrodes have also been employed with this configuration, but with a different design that allowed the rapid replacement of the electrode. The in-channel detector was based on a gold film within the separation channel. A gold-based dual electrode detector, which combined for the first time in- and end-channel detection, has been also tested. These amperometric detectors have been evaluated in combination to poly(methylmethacrylate) (PMMA) and Topas (thermoplastic olefin polymer of amorphous structure) CE-microchips. Topas is a new and promising cyclic olefin copolymer with high chemical resistance. Relevant parameters of the polymer microchip separation such as precision, efficiency or resolution and amperometric detection were studied with the different detector designs using p-aminophenol and L-ascorbic acid as model analytes in Tris-based buffer pH 9.0. PMID- 16472531 TI - Determination of unbound docetaxel and paclitaxel in plasma by ultrafiltration and liquid chromatography-tandem mass spectrometry. AB - This work describes the development of a straightforward method for the determination of free docetaxel and paclitaxel in plasma. The separation of bound and unbound drug was performed with ultrafiltration. Different ultrafiltration devices were evaluated, especially regarding non-specific binding to the device. The most appropriate device for this application was selected and a procedure to counteract non-specific binding to the ultrafiltrate collection cup was developed. This consisted of a wash procedure with methyl t-butyl ether. A liquid/liquid extraction with methyl t-butyl ether was performed and samples were analysed with a previously developed liquid chromatography-tandem mass spectrometry procedure. The method used a Merck Purospher Star RP-18 column (55 mm x 2.0 mm, 3-microm particle size) and electrospray in the positive mode. A triple quadrupole instrument was used to monitor MRM transitions. Small modifications to this procedure were made to ensure adequate sensitivity. Within- and between-day reproducibility did not exceed 15% and accuracy ranged between 94.4 and 102.5%. The calibration range of the method was from 0.4 to 100 ng/ml both for paclitaxel and docetaxel. Finally, a fast and relatively simple method could be developed. PMID- 16472532 TI - Validation of a high-performance thin-layer chromatography/densitometry method for the quantitative determination of glucosamine in a herbal dietary supplement. AB - A quantitative densitometric high-performance thin-layer chromatography (HPTLC) method was developed for the determination of glucosamine in a dietary supplement containing dried extracts of the main plants traditionally used for rheumatic disorders. The HPTLC method was chosen in order to circumvent the tedious and time-consuming sample preparation steps necessarily performed before using HPLC methods when analysing complex matrixes. Glucosamine was separated from the plant extracts on a silica gel 60 F(254) HPTLC plate using a saturated mixture of 2 propanol-ethyl acetate-ammonia solution (8%) (10:10:10, v/v/v). The plates were developed vertically up to a distance of 80 mm. For visualization, the plate was dipped into a modified anisaldehyde reagent and heated at 120 degrees C for 30 min in a drying oven. Glucosamine appeared as brownish-red chromatographic zones on a colourless background. Densitometric quantification was performed at lambda = 415 nm by reflectance scanning. The HPTLC method was successfully validated by applying the novel validation protocol proposed by a commission of the "Societe Francaise des Sciences et Techniques Pharmaceutiques" (SFSTP). In the pre validation phase, the appropriate response function was determined, while in the validation phase the method showed good performance thereby fulfilling its objective of quantifying accurately. The relative standard deviations for repeatability and intermediate precision were between 4.9 and 8.6%. Moreover, the method was found to be accurate, as the two-sided 95% beta-expectation tolerance interval did not exceed the acceptance limits of 85 and 115% on the whole analytical range (800-1,200 ng of glucosamine). PMID- 16472533 TI - Proteomic analysis of rat plasma by two-dimensional liquid chromatography and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. AB - The proteomic analysis of plasma and serum samples represents a formidable challenge due to the presence of a few highly abundant proteins such as albumin and immunoglobulins. Detection of low abundance protein biomarkers therefore requires either the specific depletion of high abundance proteins using immunoaffinity columns and/or optimized protein fractionation methods based on charge, size or hydrophobicity. Here we describe a two-dimensional (2D) liquid chromatography separation method for the fractionation of rat plasma. In the first dimension proteins were separated by chromatofocusing according to their isoelectric point (pI). In the second dimension, proteins were further fractionated by non-porous, reversed-phase chromatography according to their hydrophobicity. The data from both separations was displayed as a 2D protein expression map of pI versus retention time (relative hydrophobicity). Both separations were carried out on the ProteomeLab PF 2D system (Beckman Coulter), an instrument platform that provides a high degree of automation and real-time monitoring of the separation process. The reproducibility of the first-dimension separation was evaluated in terms of pH gradient formation. The second-dimension separation was evaluated in terms of peak retention times on the reversed-phase column. We found in four consecutive chromatofocusing separations that the pH gradient differed by less than 0.2 pH units at any time during the elution step. Second dimension retention times of peaks from identical pI fractions differed by less than 7 s in six consecutive separations. Each 2D separation generated a total of 540 fractions which were analyzed by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS). We detected approximately 275 peptides and proteins with molecular masses ranging from 3 to 225 kDa. Most fractions were found to contain multiple low and high molecular weight proteins. Differential display of 2D protein expression maps from retinol sufficient and -deficient rat plasma samples identified a fraction with several proteins that appeared to be down-regulated in the vitamin A-deficient animal. Quantitative proteomic analysis of complex samples such as plasma is still a difficult task. We discuss the potential of this approach for biomarker discovery and address the experimental challenges that remain. PMID- 16472534 TI - Qualitative and quantitative determination of extractives in heartwood of Scots pine (Pinus sylvestris L.) by gas chromatography. AB - A method for quantitative determination of extractives from heartwood of Scots pine (Pinus sylvestris L.) using gas chromatography (GC) with flame ionization detection (FID) was developed. The limit of detection (LOD) was 0.03 mg/g wood and the linear range (r = 0.9994) was up to 10 mg/g with accuracy within +/- 10% and precision of 18% relative standard deviation. The identification of the extractives was performed using gas chromatography combined with mass spectrometry (GC-MS). The yields of extraction by Soxhlet were tested for solid wood, small particles and fine powder. Small particles were chosen for further analysis. This treatment gave good yields of the most important extractives: pinosylvin, pinosylvin monomethyl ether, resin acids and free fatty acids. The method is used to demonstrate the variation of these extractives across stems and differences in north-south direction. PMID- 16472535 TI - Chemical composition of the essential oil of Salvia microstegia Boiss. et Balansa growing wild in Lebanon. AB - The essential oil of aerial parts of Salviamicrostegia Boiss. et Balansa (Lamiaceae) growing wild in Lebanon was obtained by hydrodistillation and was analysed by GC and GC-MS. 70 compounds, representing 94.0% of the oil, were identified. The major components were caryophyllene oxide (6.2%), pulegone (5.7%), 4-vinylguaiacole (5.3%), hexadecanoic acid (5.1%) and menthone (4.9%). PMID- 16472536 TI - Fast, high-efficiency peptide separations on a 50-microm reversed-phase silica monolith in a nanoLC-MS set-up. AB - Proteomic studies have stimulated the development of novel stationary phases in miniaturised chromatographic columns that permit high linear flow velocities and exhibit high resolving power. In this work, a 50-microm reversed-phase silica based monolith was chromatographically characterised for its use in proteomics applications using a nanoLC-MS set-up. It showed high efficiency for the separation of tryptic peptides under isocratic elution conditions (HETP(min)=5-10 microm at 2.4 mm/s). Flow rates up to 1.95 microL/min (18.4 mm/s) and gradient slopes up to an unusually fast 9% could be used. This resulted in rapid separations of peptide mixtures, with peak widths at half height of between 5 and 10 s. The 50-microm monolithic column was used to analyse depleted serum from a cervical cancer patient at a throughput of one sample per 30 min. PMID- 16472537 TI - Optimization of a trypsin-bioreactor coupled with high-performance liquid chromatography-electrospray ionization tandem mass spectrometry for quality control of biotechnological drugs. AB - The optimization of a silica-based trypsin bioreactor and its use in the quality control of biotechnological drugs like peptides and proteins is described. Five bioreactors based on monolithic material have been prepared, with different amount of bound trypsin. The performances of these bioreactors were compared to the proteolytic activity of a bioreactor based on silica material. The trypsin based chromatographic columns were coupled on-line with an LC/ESI/MS/MS system for digestion and identification of proteins. First, human serum albumin has been used as test protein to compare the ability of the bioreactors to hydrolyse high molecular-weight proteins. The best chromatographic material (epoxy monolithic silica) and the optimum amount of enzyme bound (7.13 mg) have been identified to obtain the highest protein recovery and an analytical reproducibility of the whole digestion, separation and identification process. The optimized enzyme reactor has been used for the on-line digestion of some biotechnological drugs such as somatotropin. Somatotropin for parentheral use has been analyzed, without sample pre-treatment, with both an on-line procedure and the traditional off-line procedure described in the European Pharmacopoeia. It was found that the cleavage efficiency (aminoacidic recovery, %AA) achieved within minutes by the developed protocol is at least comparable or even better than the conventional 4h consuming method. PMID- 16472538 TI - Determination of carbosulfan and its metabolites in oranges by liquid chromatography ion-trap triple-stage mass spectrometry. AB - Liquid chromatography ion-trap mass spectrometry LC-MSn has been successfully applied to identify and confirm carbosulfan and seven of its metabolites in oranges after pressurized liquid extraction (PLE) with dichloromethane. Mass spectra of carbosulfan and its metabolites were investigated using multiple stages of mass spectrometry. Although interpretation of the fragmentation pathways, based on mass spectra, enables structural elucidation and identification of these compounds, the proposed fragmentation pathways and ion structures need verification by exact mass measurements. The analytical method- PLE and LC-MS3 --was validated: limits of quantification (LOQ) ranged from 0.01 to 0.07 mg kg(-1); at this level, recoveries were 55-90% with RSDs (five replicate analyses) from 8 to 19%. The degradation of carbosulfan was determined in a laboratory study carried out in mature oranges (Valencia Late) and tangerines (Clementine of Nules) already harvested. Main degradation products of carbosulfan were carbofuran, 3-hydroxycarbofuran, 3-ketocarbofuran and dibutylamine. PMID- 16472539 TI - Direct enrichment and high performance liquid chromatography analysis of ultra trace Bisphenol A in water samples with narrowly dispersible Bisphenol A imprinted polymeric microspheres column. AB - Direct injection, enrichment and high performance liquid chromatography (HPLC) analysis of ultra-trace Bisphenol A (BPA) in water samples using one narrowly dispersible BPA imprinted polymeric microspheres (MIPM) column in one analysis process was developed. One BPA imprinted MIPM that had the best globular morphology and imprinted efficiency was used as HPLC stationary phase and applied to direct analysis of ultra-trace BPA in water. The optimum direct analysis conditions were: conditioning the MIPM column with water for 10min, injecting 40mL water sample directly, eluting with 70% methanol for 13min and then 100% methanol for 7min. Under the optimum conditions, the MIPM column can simultaneously extract, enrich, separate and determine ultra-trace BPA in one analysis process with UV detector by injection of large volume water samples (40mL). The calibration graph was linear with R(2)>0.998 in the ranges from 0.1 100nmol/L BPA standard solution. The intra- and inter-day RSD are less than 9.5 and 9.6%, respectively. The limit of quantification was 0.1nmol/L. RSD for spiked tap and lake water was less than 8.9% and the recoveries were 96-101.8%. The enrichment factor for BPA was 10,000 as 40mL water sample was directly injected and analyzed. PMID- 16472540 TI - Chromatographic fingerprint analysis--a rational approach for quality assessment of traditional Chinese herbal medicine. AB - Traditional Chinese Herbal Medicine (TCHM) contain multiple botanicals, each of which contains many compounds that may be relevant to the medicine's putative activity. Therefore, analytical techniques that look at a suite of compounds, including their respective ratios, provide a more rational approach to the authentication and quality assessment of TCHM. In this paper we present several examples of applying chromatographic fingerprint analysis for determining the identity, stability, and consistency of TCHM as well as the identification of adulterants as follows: (1) species authentication of various species of ginseng (Panax ginseng, Panax quinquefolium, Panax noto-ginseng) and stability of ginseng preparations using high performance thin-layer chromatography (HPTLC) fingerprint analysis; (2) batch-to-batch consistency of extracts of Total Glycosides of Peony (TGP), to be used as a raw material and in finished products (TGP powdered extract products), using high performance liquid chromatography (HPLC) fingerprint analysis with a pattern recognition software interface (CASE); (3) documenting the representative HPLC fingerprints of Immature Fruits of Terminalia chebula (IFTC) through the assessment of raw material, in-process assay of the extracts, and the analysis of the finished product (tablets); (4) HPLC fingerprint study demonstrating the consistent quality of total flavonoids of commercial extracts of ginkgo (Ginkgo biloba) leaves (EGb) along with detection of adulterations. The experimental conditions as well as general comments on the application of chromatographic fingerprint analysis are discussed. PMID- 16472541 TI - Vacancy ion-exclusion chromatography of inorganic acids on a weakly acidic cation exchange resin column. AB - Vacancy ion-exclusion chromatography (VIEC) for inorganic acids such as H(2)SO(4), HCl, H(3)PO(4), HNO(3), HI and HF is tested on a polymethacrylate based weakly acidic cation-exchange resin column in the H(+)-form. That is, mixture of inorganic acids in the mobile phase is adsorbed to the resin phase passing through the separation column, and each vacant peak induced by injecting water is determined. Retention times are dependent on the degrees of retention for each analyte in the resin phase. In VIEC, well-shaped peaks of inorganic acids are produced, leading to efficient separations. However, retention behaviors of inorganic acids were strongly affected by the concentrations of the acids in the mobile phase. Sulfosalicylic acid was mixed with inorganic acids in the mobile phase prior to the introduction of a separation column in order to obtain the well-resolutions in the lower concentrations of the acids. By using this method, the separations of inorganic acids could be achieved in the range of 0.01-1 mM, and the linear ranges could be extended over two-orders of magnitude. This is considered since the protonated carboxylic groups fixed on the resin phase were increased with increasing the acid concentrations in the mobile phase, and the penetration effects for the acids to the resin phase were thus enhanced. The detection limits (S/N=3) were below 1.0 microM for all analyte acids. Precision values for retention times were below 0.32% and for peak area were below 0.91%. PMID- 16472542 TI - 16S rRNA gene probe quantitates residual host cell DNA in pharmaceutical-grade plasmid DNA. AB - The development and widespread use of DNA-based vaccination against infectious pathogens have been a great triumph of medical science. Quality control of DNA vaccines as biopharmaceutical productions is a problem to solve. Residual genomic DNA of engineering bacteria has been identified as a potential risk factor, so whose level must be controlled under the regulatory standards. We report a dot blot hybridization method to detect residual host cell DNA in purified DNA vaccines. The assay utilizes PCR amplified and digoxigenin-labeled Escherichia coli 16S rRNA gene as probe. The sensitivity of the dot-blot hybridization assay with E. coli 16S rRNA gene probe was evaluated in comparison with single copy UidR gene probe. The optimized dot-blot hybridization assay had both low background and a suitable sensitivity, detecting 10 pg of residual E. coli DNA. The method is suitable in the routine use of measuring the levels of residual E. coli DNA in the pharmaceutical-grade DNA vaccine. PMID- 16472543 TI - Studies on in vitro expression and in vivo immunogenicity of a recombinant MVA HIV vaccine. AB - Here we conduct dose-response studies for in vitro expression and in vivo immunogenicity for a recombinant modified vaccinia Ankara (MVA) vaccine that expresses HIV Gag, Pol, and Env proteins. The dose-response studies for in vitro expression used fluorescent-activated cell sorting to score Gag- and Env expressing cells and showed good increases for antigen expression with increasing MVA dose. In these studies, a 1000-fold increase in the dose of MVA resulted in a 300-fold increase in the frequency of antigen-expressing cells. In contrast, dose response studies for in vivo immunogenicity showed <10-fold increases in elicited T cells and Ab for 100-1000-fold increases in the dose of inoculated MVA. These studies used intracellular cytokine assays to enumerate responding CD8 and CD4 T cells and an ELISA to score anti-Env Ab. The shallow dose-response curves for immunogenicity were observed post priming as well as post boosting of an MVA or a DNA prime. PMID- 16472544 TI - Serum-free influenza virus production avoiding washing steps and medium exchange in large-scale microcarrier culture. AB - A complete serum-free process without washing steps and medium exchange before infection for influenza A virus vaccine production (equine and human) is described for cultivation in roller bottles and in a 5-L stirred tank microcarrier system. Adherent Madin-Darby canine kidney cells (MDCK) were adapted from growth in serum containing GMEM medium to growth in serum-free Ex-Cell MDCK medium. Roller bottle experiments showed that the medium exchange step, typically required for serum containing vaccine production processes, could be omitted without losses in virus titre and without limitations in glucose or glutamine supply in the cultivation medium. The serum-free medium could even be used glutamine-free as it contained pyruvate, resulting in very low levels of ammonia. Cell attachment onto microcarriers was critical. Therefore, microcarriers had to be preconditioned in medium. Also, trypsin concentration used for inoculum preparation had to be reduced. After these modifications 1.3 x 10(6)cells/mL were obtained after 97 h (2g/L Cytodex 1) of cell growth. Maximum virus titres of 2.3 2.9 log HA units/100 microL were obtained from infections with a multiplicity of infection (moi) of 0.05-0.10 for human and equine influenza A virus. Metabolite and amino acid profiles as well as on-line data for the serum-free process are compared with the serum containing process. Omission of the medium exchange before infection clearly simplified the process and reduced sterility risks due to washing steps. PMID- 16472545 TI - An improved rearranged Human Papillomavirus Type 16 E7 DNA vaccine candidate (HPV 16 E7SH) induces an E7 wildtype-specific T cell response. AB - A new and very promising approach in vaccine development is the application of naked DNA. In comparison to conventional vaccines it offers several advantages, especially if there is a need for the development of low cost vaccines. Infection with high-risk human papillomaviruses (hr-HPVs) is the major risk factor for the development of cervical cancer (cc), the third most common cancer in women worldwide. The HPV E7 oncogene is constitutively expressed in HPV-infected cells and represents an excellent target for immune therapy of HPV-related disease. Therefore, we chose the HPV-16 E7 as model antigen in the development of a therapeutic DNA vaccine candidate. For safety reasons the use of a transforming gene like the HPV-16 E7 for DNA vaccination is not feasible in humans. In consequence we have generated an artificial ("shuffled") HPV-16 E7-gene (HPV-16 E7SH), containing all putative cytotoxic T-lymphocyte (CTLs) epitopes and exhibiting high safety features. Here, we show the induction of a strong E7 wildtype (E7WT) directed cellular and humoral immune response including tumor protection and regression after in vivo immunization in the murine system. Moreover, the vaccine candidate demonstrated immunogenicity in humans, demonstrated by priming of antigen-specific T cells in vitro. Importantly, the artificial HPV-gene has completely lost its transforming properties as measured in soft agar transformation assays. These results may be of importance for the development of vaccines based on oncogenes or oncoproteins. PMID- 16472547 TI - A MUC1/IL-18 DNA vaccine induces anti-tumor immunity and increased survival in MUC1 transgenic mice. AB - MUC1 (mucin 1) is a tumor-associated antigen that is overexpressed in many adenocarcinomas. Active immunotherapy targeting tumors expressing MUC1 could have great treatment value. MUC1 DNA vaccines were evaluated in MUC1 transgenic (MUC1.Tg) mice challenged with MC38/MUC1+ tumor cells. Vaccination with MUC1 plasmid DNA (pMUC1) alone was insufficient to induce tumor protection. However, co-administration of pMUC1 with a plasmid encoding murine interleukin-18 (pmuIL 18) resulted in significant tumor protection and survival after tumor challenge. Protection was durable in the absence of additional vaccination, as demonstrated by continued protection of vaccinated mice following tumor rechallenge. Mice surviving challenges with MC38/MUC1+ cells showed significant protection after challenge with MUC1(-) MC38 tumor cells, suggesting that these mice had developed immune responses to epitopes shared between the tumor cell lines. Antibody mediated depletion of lymphocyte subsets demonstrated that protection was due largely to CD4+ T cells. This work demonstrates that a naked DNA vaccine can break tolerance to MUC1 and induce an immune response capable of mediating both significant protection from tumor challenge and increased survival. PMID- 16472546 TI - A truncated C-terminal fragment of Mycobacterium tuberculosis HSP70 gene enhanced potency of HBV DNA vaccine. AB - DNA vaccine represents an attractive approach to therapy of chronic hepatitis B virus (HBV) infection because of its ability to generate antigen-specific immunity; nevertheless, there is still a need to increase the potency of DNA vaccine. Mycobacterium tuberculosis heat shock protein70 (HSP70) has both chaperon and cytokine functions, and has been shown to act as an adjuvant when co administered with peptide antigens or given as fusion proteins. Here we evaluated the effects of two truncated HSP70 molecules, N-terminal domain (HSP70(1-360), amino acids 1-360) and C-terminal domain (HSP70(359-610), amino acids 359-610) of mycobacterial HSP70, on the potency of antigen-specific immunity generated by a HBV DNA vaccination. We found that only the HSP70(359-610)-fused HBV DNA vaccination resulted in a significant increase in hepatitis B surface antigen (HBsAg)-specific humoral response, while the HSP70(1-360)- or the complete HSP70 molecule-fused vaccine did not. Moreover, HSP70(359-610)-fused DNA vaccine did not induce anti-HSP70 antibody. Interestingly, HSP70(359-610) not only enhanced HBsAg-specific cytotoxic lymphocytes (CTL) responses but also overcame the epitope suppression caused by L(d)-restricted epitope. Meanwhile, HSP70(369-610) mediated T helper (Th) cell balance towards Th1 pathway. In a HBV transgenic mouse model, the HSP70(359-610) fusion vaccine facilitated clearance of circulating HBsAg and down-regulation of HBV replication. These results suggested that the truncated mycobacterial HSP70 molecule, HSP70(359-610), might be a superior candidate to deliver the adjuvant function in HBV DNA vaccination instead of the complete HSP70 molecule. PMID- 16472548 TI - Response of steroid-treated former preterm infants to a single dose of meningococcal C conjugate vaccine. AB - Attenuated antibody responses have been reported in preterm infants who received neonatal dexamethasone treatment. The duration of immunosuppression may extend into later infancy. This study assessed the immune response of former preterm infants to a single meningococcal serogroup C conjugate (MCC) immunisation given after infancy. A cohort of 49 toddlers born at less than 33 weeks' gestation were given an initial dose of MCC vaccine at a median age of 13 months; 11 had received dexamethasone in the neonatal period. Sera obtained 4 weeks post immunisation were analysed for serum bactericidal antibody (SBA) and serogroup C specific IgG antibody concentrations. Immune responses were compared with those of an historical cohort of 70 term toddlers given a single dose of the same vaccine at age 13 months. An SBA titre of > or =8 was taken to indicate a protective response. Following a single MCC dose, the SBA geometric mean titre (GMT) for former preterm infants was 249 (95% C.I. 111, 558), not significantly different from that of the historical term cohort whose SBA GMT was 141 (95% C.I. 89, 224) (p=0.06). A significantly lower proportion of former preterm infants achieved a protective SBA titre of > or =8 compared with term infants, 37/48 (77%) versus 64/70 (91%), (p=0.03). For steroid-treated and non steroid-treated subgroups, SBA GMTs were 1237 (95% C.I. 250, 6132) and 154 (62, 385), respectively, and numbers achieving an SBA titre of > or =8 were 10/11 (91%) and 27/37 (73%), (p=0.42). Most children born at <33 weeks' gestation mount a protective immune response to a single MCC vaccine dose given at age 13 months, but fewer former preterm infants attain a protective SBA titre of 8 compared with term infants. Previous neonatal dexamethasone treatment does not appear to attenuate immune response after infancy. PMID- 16472549 TI - Differential regulation of the CXCR2 chemokine network in rat brain trauma: implications for neuroimmune interactions and neuronal survival. AB - Chemokine receptors represent promising targets to attenuate inflammatory responses and subsequent secondary damage after brain injury. We studied the response of the chemokines CXCL1/CINC-1 and CXCL2/MIP-2 and their receptors CXCR1 and CXCR2 after controlled cortical impact injury in adult rats. Rapid upregulation of CXCL1/CINC-1 and CXCL2/MIP-2, followed by CXCR2 (but not CXCR1), was observed after injury. Constitutive neuronal CXCR2 immunoreactivity was detected in several brain areas, which rapidly but transiently downregulated upon trauma. A second CXCR2-positive compartment, mainly colocalized with the activated microglia/macrophage marker ED1, was detected rapidly after injury in the ipsilateral cortex, progressively emerging into deeper areas of the brain later in time. It is proposed that CXCR2 has a dual role after brain injury: (i) homologous neuronal CXCR2 downregulation would render neurons more vulnerable to injury, whereas (ii) chemotaxis and subsequent differentiation of blood-borne cells into a microglial-like phenotype would be promoted by the same receptor. PMID- 16472550 TI - Body posture facilitates retrieval of autobiographical memories. AB - We assessed potential facilitation of congruent body posture on access to and retention of autobiographical memories in younger and older adults. Response times were shorter when body positions during prompted retrieval of autobiographical events were similar to the body positions in the original events than when body position was incongruent. Free recall of the autobiographical events two weeks later was also better for congruent-posture than for incongruent posture memories. The findings were similar for younger and older adults, except for the finding that free recall was more accurate in younger adults than in older adults in the congruent condition. We discuss these findings in the context of theories of embodied cognition. PMID- 16472551 TI - Effect of chlorpyrifos-methyl on steroid and thyroid hormones in rat F0- and F1 generations. AB - Chlorpyrifos-methyl (CPM) suppressed androgenic activity in Hershberger assay using castrated rats. Acute oral lowest-observed-adverse-effect-level (LOAEL) and no-observed-adverse-effect-level (NOAEL) was evaluated as 12 and 0.1 mg/kg bw, respectively, based on its major effect of cholinesterase inhibition. Also, repeated oral NOAEL was 0.1 mg/kg bw/day based on adrenal damage in rats. We investigated one-generation reproductive toxicity of CPM focusing on endocrine disrupting effects by the administration of 1, 10 and 100 mg/kg bw/day CPM to mature SD rats (F0) through pre-mating, mating, gestation and lactation period and to their offspring (F1) until 13 weeks age via gavage. A group treated with corn oil served as vehicle control. In F0 rats, the most affected organs were adrenal glands as increased in weight at all doses of CPM in males and at 10 and 100 mg/kg CPM in females and adrenal vacuolation at CPM 10 and 100 mg/kg. The relative and absolute ovaries and the absolute seminal vesicle weights were decreased but the weights of liver, spleen or kidneys were increased at 100 mg/kg CPM. Parameters representing reproductive performances as mating ratio, gestation length and delivery index were not affected, except for decreased fertility index and numbers of implantation and born pups and a higher male sex ratio of pups at CPM 100 mg/kg. F1 pups exposed to CPM 100 mg/kg in utero and via maternal milk showed lower body weight with changes of absolute or relative weights of brain, ovary, liver, spleen and epididymis and decreased absolute not relative anogenital distance at weanling time. The time of vaginal patency and preputial separation and estrous cycling pattern of F1 rats were not impacted by CPM. After further 10 weeks oral administration until 13 weeks old, adrenal glands, brain, liver, spleen or kidneys tended to be increased, while thyroid gland, testes and ventral prostate of F1 male rats were decreased at CPM 10 or 100 mg/kg. Histopathologically, necrosis or vacuolation of thyroid follicular epithelial cells and adrenal cortical cells were observed at all doses of CPM. Serum levels of estradiol, testosterone, T4 and T3 were significantly lower while TSH and cholesterol were higher in both F1 female and male rats treated with CPM though dose-responsiveness was not clear in F1 females. Decreased sperm were counted in F1 rats at CPM 100 mg/kg. As a whole, LOAEL and NOAEL was evaluated as 10 and 1 mg/kg bw, respectively, based on decreased estradiol and T4 and increased TSH in serum of F1 male rats, and when considering histopathological alteration of adrenal and thyroid glands, LOAEL assumed to be lower than 1 mg/kg bw. This study elucidates that CPM exhibit weak reproductive toxicity in F0 rats exposed at adulthood and negligible effects in F1 offspring exposed in utero and via lactation at weanling, but induce anti-androgenic effect and hypothyroidism after long term exposure from in utero through sexual maturation of F1 rats. PMID- 16472552 TI - Expression of Ku80 correlates with sensitivities to radiation in cancer cell lines of the head and neck. AB - The Ku protein is essential for the repair of a majority of DNA double-strand breaks in mammalian cells. The purpose of this study was to investigate the relationship between the expression of Ku70/80 and sensitivity to radiation in cancer cell lines of the head and neck. The sensitivity to radiation in various head and neck cancer cell lines (AMC-HN-1 to -9) was analyzed by colony forming assay. Of the nine cell lines examined, the most radiosensitive cell line (AMC-HN 3) and the most radioresistant cell line (AMC-HN-9) were selected for this experiments. The expression of Ku70/80 was examined after irradiation using real time PCR, Western blotting and immunofluorescence in two different cell lines. Cell cycle distribution after irradiation were analysed. A differential radioresponse was demonstrated by expression of Ku70/80 in AMC-HN-3 and AMC-HN-9 cells. While the expression of Ku70 was slightly increased in the radioresistant AMC-HN-9 cell line, the expression of Ku80 was remarkably increased, suggesting a correlation between Ku80 expression and radiation resistance. Overexpression of Ku80 plays an important role in the repair of DNA damage induced by radiation. Ku80 expression may provide an effective predictive assay of radiosensitivity in head and neck cancers. PMID- 16472554 TI - Reducing hospital-acquired infection by design: the new University College London Hospital. AB - The planning and building of new University College London Hospital has provided a challenge to the Infection Control Team to ensure all areas are easy to clean, air flows are correct and appropriate and that there is every opportunity for handwashing. The seven year process illustrates the paper by Stockley et al. ['Stockley JM, Constantine CE, Orr KE, The Association of Medical Microbiologists' New Hospital Developments Project Group. Building new hospitals: a UK infection control perspective. PMID- 16472553 TI - 15-Lipoxygenase-1 induced by interleukin-4 mediates apoptosis in oral cavity cancer cells. AB - It is unknown whether IL-4 induces apoptosis in oral cavity cancer (OCC) cells and, if so, what the mediator of this apoptosis is. Therefore, this study investigated whether apoptosis of OCC cells is induced by IL-4 and whether 15 lipoxygenase-1 (15-LO-1), induced by IL-4, is the mediator of this apoptosis. SCC 1483 oral cavity cancer cells were used in these experiments, and flow cytometry and PARP cleavage were used to examine apoptosis. At an IL-4 dosage that inhibited 50% of cell proliferation, apoptosis was observed, and this apoptosis was inhibited by 2.2 microM caffeic acid (CA). 15-LO-1 mRNA expression was observed beginning at 8 h after treatment with IL-4, and apoptosis increased after 24 h treatment with IL-4. In this apoptosis, the caspase cascade was not involved. In summary, IL-4 induced apoptosis in SCC 1483 OCC cells, and 15-LO-1, induced by IL-4, may mediate this apoptotic pathway. PMID- 16472555 TI - Thermodynamics of membrane domains. AB - The concept of lipid rafts and the intense work toward their characterization in biological membranes has spurred a renewed interest in the understanding of domain formation, particularly in the case of cholesterol-containing membranes. The thermodynamic principles underlying formation of domains, rafts, or cholesterol/phospholipid complexes are reviewed here, along with recent work in model and biological membranes. A major motivation for this review was to present those concepts in a way appropriate for the broad readership that has been drawn to the field. Evidence from a number of different techniques points to the conclusion that lipid-lipid interactions are generally weak; therefore, in most cases, massive phase separations are not to be expected in membranes. On the contrary, small, dynamic lipid domains, possibly stabilized by proteins are the most likely outcome. The results on mixed lipid bilayers are used to discuss recent experiments in biological membranes. The clear indication is that proteins partition preferentially into fluid, disordered lipid domains, which is contrary to their localization in ordered, cholesterol/sphingomyelin rafts inferred from detergent extraction experiments on cell membranes. Globally, the evidence appears most consistent with a membrane model in which the majority of the lipid is in a liquid-ordered phase, with dispersed, small, liquid-disordered domains, where most proteins reside. Co-clustering of proteins and their concentration in some membrane areas may occur because of similar preferences for a particular domain but also because of simultaneous exclusion from other lipid phases. Specialized structures, such as caveolae, which contain high concentrations of cholesterol and caveolin are not necessarily similar to bulk liquid-ordered phase. PMID- 16472556 TI - A small HSP, Lo18, interacts with the cell membrane and modulates lipid physical state under heat shock conditions in a lactic acid bacterium. AB - The small heat shock proteins (sHSP) are characterized by a chaperone activity to prevent irreversible protein denaturation. This study deals with the sHSP Lo18 induced by multiple stresses in Oenococcus oeni, a lactic acid bacterium. Using in situ immunocytochemistry and cellular fractionation experiments, we demonstrated the association of Lo18 with the membrane in O. oeni cells submitted to heat shock. The same result was obtained after exposure of cells to ethanol or benzyl alcohol, agents known to have an influence on membranes. For the different stresses, the protein was located on the periphery of the cell at membrane level and was also found within the cytoplasm. In order to determine if Lo18 could interact with the phospholipids, we used model membranes made of lipids extracted from O. oeni cells. Using fluorescence anisotropy of diphenylhexatriene (DPH) and generalized polarization of Laurdan, we showed that purified Lo18 interacts with these liposomes, and increases the molecular order of the lipid bilayer in these membranes when the temperature reaches 33.8 degrees C. All these data suggest that Lo18 could be involved in an adaptive response allowing the maintenance of membrane integrity during stress conditions in O. oeni cells. PMID- 16472557 TI - Effects of pH-induced variations of the charge of the transmembrane alpha-helical peptide Ac-K2(LA)12K2-amide on the organization and dynamics of the host dimyristoylphosphatidylcholine bilayer membrane. AB - The effects of the transmembrane alpha-helical peptide Ac-K(2)(LA)(12)K(2)-amide ((LA)(12)) on the phase transition and dynamics of saturated dimyristoylphosphatidylcholine (DMPC) membranes were investigated at different pH using conventional and saturation-recovery EPR observations of phosphatidylcholine spin labels. At a peptide-to-DMPC ratio of 1/10, the main phase-transition temperature of the DMPC bilayer is decreased by 4.0 degrees C when measured at pH 7.0, by 1.6 degrees C when measured at pH 9.5, and not affected when measured at pH 11.5. This reversible pH effect is due to the subsequent neutralization of the positive charges of lysine side chains at both ends of (LA)(12). Apparent pK(a)s of the lysine side chain amino groups of (LA)(12) in DMPC bilayer are 8.6 and approximately 10.9, as compared with the pK(a) value of 10.5 for these groups when lysine is dissolved in water. Saturation-recovery curves as a function of oxygen concentration using phosphatidylcholine spin labels in DMPC bilayer containing (LA)(12) are always mono-exponential when measured at pH 7.0 and 9.5. This observation is consistent with the hypothesis that the lipid exchange rates among the bulk, boundary, and (LA)(12)-rich regions are faster than 0.5 micros, the electron spin-lattice relaxation time in the presence of molecular oxygen, suggesting that stable oligomers of (LA)(12) do not form. Neutralization of one lysine side chain positive charge on each end of the peptide significantly decreases the ordering effect of (LA)(12) on the lipid hydrocarbon chains, while its effect on the reorientational motion of terminal groups of lipid hydrocarbon chains is rather moderate. It does not affect the local diffusion-solubility product of oxygen measured in the DMPC-(LA)(12) membrane interior. PMID- 16472559 TI - Contraception: an international perspective. AB - Although it is estimated that the population growth rate will decline to a replacement level by 2050, it is also now predicted that the total world population will reach 8.9 billion in that year -- far higher than the 2004 estimate of 6.4 billion. More than 26 billion new couples will need contraceptives in the next half century. Although a steady increase in contraceptive use has been observed in both developed and developing countries, the contraceptive needs of a high percentage of couples have not yet been met and the number of unplanned pregnancies continues to increase. The actual use of contraception differs from region to region. Although no new method has been registered for many years, several new products have been marketed during the last 5. Among these are new implants, medicated intrauterine systems, contraceptive vaginal rings, transdermal patches and several new combined oral contraceptive formulations. New contraceptive methods have been developed to meet the objectives of expanding contraceptive choices for both women and men and answering an unmet need for contraceptives with a long-term action that meet the expectations of consumers. Simplicity, reversibility and effectiveness are the desired features of a male contraceptive, but no new male contraceptive method is yet available. New areas of basic research include studies on genes, proteins and enzymes involved in the reproductive system. The new methods will be targeted to specific interactions within the reproductive system at the level of ovaries and testes, as well as between spermatozoa and ova. This futuristic approach still keeps in mind the need for better access to existing contraceptive methods, as well as the discovery of new contraceptives that are simple to use, safe, reversible and inexpensive. In the future, contraceptives may be combined with other medicinal agents to provide dual protection against both pregnancy and other preventable conditions, such as sexually transmitted infections. PMID- 16472558 TI - Effect of triorganotin compounds on membrane permeability. AB - Organotin compounds are widely distributed toxicants. They are membrane-active molecules with broad biological toxicity. In this contribution, we study the effect of triorganotin compounds on membrane permeability using phospholipid model membranes and human erythrocytes. Tribultyltin and triphenyltin are able to induce the release of entrapped carboxyfluorescein from large unilamellar vesicles. The rate of release is similar for phosphatidylcholine and phosphatidylserine systems and the presence of equimolar cholesterol decreases the rate of the process. Release of carboxyfluorescein is almost abolished when a non-diffusible anion like gluconate is present in the external medium, and it is restored by addition of chloride. Tributyltin is able to cause hemolysis of human erythrocytes in a dose-dependent manner. Relative kinetics determination shows that potassium leakage occurs simultaneously with hemoglobin release. Hemolysis is reduced when erythrocytes are suspended in a gluconate medium. These results indicate that triorganotin compounds are able to transport organic anions like carboxyfluorescein across phospholipids bilayers by exchange diffusion with chloride and suggest that anion exchange through erythrocyte membrane could be related to the process of hemolysis. PMID- 16472561 TI - Safety and efficacy of an extended-regimen oral contraceptive utilizing continuous low-dose ethinyl estradiol. AB - OBJECTIVE: To assess the efficacy and safety of Seasonique, a 91-day extended regimen oral contraceptive (OC) utilizing continuous low-dose ethinyl estradiol (EE) during the typical hormone-free interval. METHODS: A multicenter, open label, 1-year study of Seasonique [30 microg EE/150 microg levonorgestrel (LNG)] for 84 days followed by EE 10 microg for 7 days was conducted in sexually active, adult women of childbearing potential. All patients completed daily electronic diaries to monitor compliance and bleeding. RESULTS: Method failure rate was 0.78 (Pearl index) and 0.64% (life table analysis). Cycle control and safety of the regimen were similar to that reported for other OCs. CONCLUSION: This study demonstrates that Seasonique is effective, safe and well tolerated for the prevention of pregnancy. PMID- 16472560 TI - Risk of nonfatal venous thromboembolism in women using a contraceptive transdermal patch and oral contraceptives containing norgestimate and 35 microg of ethinyl estradiol. AB - CONTEXT: There is concern that a new transdermal contraceptive patch containing ethinyl estradiol (EE) and the progestin norelgestromin increases the risk for venous thromboembolism (VTE) compared to previously marketed oral contraceptives (OCs). OBJECTIVE: Quantitative information was obtained on the risk of nonfatal VTE in women using the contraceptive patch in comparison to women using OCs, norgestimate (either monophasic or triphasic) and 35 microg EE (norgestimate-35), an OC that has been marketed for over a decade. DESIGN, SETTING AND PARTICIPANTS: Nested case-control design based on information from PharMetrics, a US-based company that collects and organizes information on claims paid by managed care plans. The study was nested among all women aged 15 to 44, who started either the contraceptive patch or norgestimate-35 after April 1, 2002. Cases were women with current use of one of these two study drugs and a documented diagnosis of VTE in the absence of identifiable clinical risk factors (idiopathic VTE). Up to four controls were matched to each case by age and calendar time. MAIN OUTCOME MEASURES: Odds ratios (ORs) comparing the risk of nonfatal VTE in new users of the two contraceptives and incidence rates of nonfatal VTE for new users of each of the study contraceptives. RESULTS: We identified 68 newly diagnosed, idiopathic cases of VTE in the study population. In the case-control analysis, the OR comparing the contraceptive patch to norgestimate-35 was 0.9 (95% CI 0.5 1.6). The overall incidence rate for VTE was 52.8 per 100,000 women-years (95% CI 35.8-74.9) among users of the contraceptive patch and 41.8 per 100,000 women years among users of norgestimate-35 (95% CI 29.4-57.6), and the age-adjusted VTE incidence rate ratio (IRR) for current use of the contraceptive patch vs. norgestimate-35 was 1.1 (95% CI 0.7-1.8). CONCLUSIONS: The risk of nonfatal VTE for the contraceptive patch is similar to the risk for OCs containing 35 microg ethinylestradiol and norgestimate. PMID- 16472562 TI - Ovarian follicular dynamics during conventional vs. continuous oral contraceptive use. AB - OBJECTIVES: The purpose of this study was to characterize ovarian follicular and endometrial development during conventional vs. continuous oral contraceptive (OC) dosing regimens, to explore follicular development during the hormone-free interval (HFI) and to examine follicular development following OC discontinuation. STUDY METHODS: A randomized clinical trial involving 36 clinically normal healthy women between the ages of 18 and 35 years (24.4 +/- 3.9, SEM). Transvaginal ultrasonography and blood sampling were done to ascertain ovarian function. RESULTS: Fewer follicles > 4 mm developed during continuous vs. conventional OC use (p = .006). No dominant follicles developed during continuous OC use vs. eight dominant follicles (16.1 +/- 3.3 mm) during the conventional OC regimen. Two of eight (25%) dominant follicles ovulated. All dominant follicles began development during the HFI. Following discontinuation of OC use, ovulation took approximately 5 days longer when compared to natural cycles. CONCLUSION: Continuous OC regimens more effectively prevent dominant follicle development and breakthrough ovulation. The slight delay in time to ovulation following OC discontinuation and natural cycles could be attributed to suppression of follicle wave activity. PMID- 16472563 TI - Termination of pregnancy in patients with previous cesarean section. AB - OBJECTIVE: To evaluate the safety and efficacy of termination of pregnancy using mifepristone and misosprostol at more than 15 weeks' gestation in patients with uterine scar due to previous cesarean section. MATERIALS AND METHODS: This retrospective study was conducted in a tertiary maternity ward between January 2000 and October 2004. A total of 252 women at more than 15 weeks' gestation underwent termination of pregnancy including 50 women with uterine scar due to previous cesarean section (Group 1) and 202 control patients (Group 2) without known uterine scar. Abortion was induced with mifepristone and a prostaglandin analogue. Women between 15 and 34 weeks' gestation received misoprostol intravaginally every 3 h at doses of 200 microg (Group 1) or 400 microg (Group 2). Women at more than 34 weeks' gestation received Prostin E2 vaginal gel. Main end points were hemorrhage, fever, retained placenta, occurrence of complications including uterine rupture and dehiscence, and final outcome. RESULTS: A total of 13 (26%) patients in Group 1 and 79 (39.1%) in Group 2 were at more than 24 weeks' gestation. The abortion failure rate was 2% (1/50) in Group 1 and 0.5% (1/202) in Group 2 (p = .28). The median induction-to-delivery interval was 8.5 h (range, 3.0-114.2 h) for Group 1 and 9.0 h (range, 1.3-124.3 h) in Group 2 (p = .26). One case of uterine rupture and one case of dehiscence were observed, both in women in Group 1. The incidence of hemorrhage was not significantly different between Group 1 and Group 2 (2% vs. 0.9%, respectively, p = .56). The incidence of retained placenta was higher in the Group 1 (70% vs. 52.5%, respectively, p = .025). CONCLUSION: In this retrospective series of women who underwent abortion at 15-35 weeks' gestation using mifepristone and a prostaglandin analogue for labor induction abortion, history of cesarean section was not associated with higher morbidity except risk of uterine rupture. However, dose and interval of misoprostol should be determined. A larger study is needed before drawing definitive conclusions about the safety of these regimens. PMID- 16472565 TI - The effect of tranexamic acid for treatment of irregular uterine bleeding secondary to Norplant use. AB - OBJECTIVE: The purpose of this study is to evaluate the effects of tranexamic acid and placebo on controlling irregular uterine bleeding secondary to Norplant use. MATERIALS AND METHODS: A prospective randomized, double-blind, placebo controlled trial was conducted. Sixty-eight Norplant users who had irregular bleeding were randomly allocated into two groups. A total of 34 users received tranexamic acid 500 mg twice a day for 5 days; the placebo was given to the other 34 users in the same manner. The total number of days of bleeding and spotting and the percentage of women whose irregular uterine bleeding was stopped was analyzed at the end of weeks 1 and 4. RESULTS: In the follow-up period (4 weeks after the initial treatment), a bleeding-free interval of > 20 days was found in 58.8% of the subjects treated with tranexamic acid and in 76.5% treated with placebo (p = .12). The percentage of subjects whose irregular uterine bleeding stopped during week 1 after the initial treatment was significantly higher in the tranexamic acid group than those in the placebo group (64.7% vs. 35.3%, p = .015). The mean number of bleeding/spotting days was not significantly different between the groups (15.4 vs. 12.7 days, p = .182). CONCLUSIONS: Tranexamic acid for 5 days appears to be an effective treatment for irregular uterine bleeding associated with Norplant use only while it is being used. However, treatment for 5 days does not seem to provide long-term benefit. PMID- 16472564 TI - Comparison of two regimens of a monthly injectable contraceptive containing dihydroxyprogesterone acetophenide and estradiol enanthate. AB - INTRODUCTION: This study compared two regimens of a monthly injectable contraceptive containing dihydroxyprogesterone acetophenide 150 mg and estradiol enanthate 10 mg (Perlutan) over 12 cycles of use. METHODS: Three hundred sixty five adolescents were randomized into two groups. The patients in Group 1 received an initial injection of Perlutan on the 1st-5th day of their menstrual cycle and subsequent injections every 30 +/- 3 days, whereas those in Group 2 followed the traditional schedule of administration in which the first injection is administered between Days 7 and 10 of their menstrual cycle and subsequent injections 7-10 days after Day 1 of withdrawal bleeding. This schedule may result in an irregularity in the timing of injections. RESULTS: No significant difference was found between the two groups regarding tolerability or pregnancy (two in Group 1 and three in Group 2). CONCLUSION: Monthly administration limits the annual number of injections to a maximum of 12, thus frequently reducing the total annual dose while maintaining efficacy and tolerability similar to those obtained with the traditional regimen. PMID- 16472566 TI - Uterine perforation in women using a levonorgestrel-releasing intrauterine system. AB - OBJECTIVE: To determine an estimated incidence of uterine perforations related to the insertion of a levonorgestrel-releasing intrauterine system (LNG IUS) and to identify possible risk factors. DESIGN: Retrospective, case report study. SETTING: Hospitals in Limburg, the Netherlands. METHODS: Gynecologists in hospitals in Limburg were asked about uterine perforations related to the insertion of a LNG IUS between 1999 and 2002. The charts of the reported perforations were studied. Data on the patient, doctor, insertion, diagnosis and removal were collected for every reported uterine perforation. RESULTS: In Limburg, the estimated incidence of uterine perforations related to the insertion of a LNG IUS is 2.6 per 1000 insertions. Insertion in lactating women, even beyond 6 weeks after delivery, was shown to be an important risk factor. CONCLUSIONS: Complete registration of complications provides a greater insight into the actual incidence of LNG IUS-related uterine perforations and their possible consequences. This may eventually lead to a decrease in complications. PMID- 16472568 TI - A qualitative study of pharmacists' perspectives on the supply of emergency hormonal contraception via patient group direction in the UK. AB - AIM: To investigate pharmacists' views and experiences of supplying emergency hormonal contraception (EHC) via a group prescribing protocol in community pharmacies in the UK. DESIGN: Qualitative study using depth interviews. SETTING: Community pharmacists in Manchester, Salford and Trafford (Greater Manchester), and Lambeth, Southwark and Lewisham (London) Health Action Zones in the UK. PARTICIPANTS: Forty-four community pharmacists supplying EHC in Manchester, Salford and Trafford, and Lambeth, Southwark and Lewisham (London). RESULTS: Pharmacists were broadly very positive about their experiences supplying EHC via the group prescribing protocol. Pharmacists identified many benefits of the EHC schemes for clients, in particular, improved access to EHC at no cost to clients. The confidential nature of the scheme was also seen as an advantage as was the scope for referral to other service providers. Pharmacists also believed that the scheme had benefits for the profession in terms of enhanced professional standing. However, their concerns included the extent of repeated use of EHC, the possible impact on contraceptive behaviors and sexually transmitted infections and its impact on male coercive sexual behavior. CONCLUSIONS: Although pharmacy supply of EHC may improve access for some clients and is perceived as a popular service, research into the implications of the schemes as identified in this study need to be conducted. PMID- 16472567 TI - Mifepristone vaginally in an early medical abortion regimen: a pilot study. AB - OBJECTIVE: This prospective observational pilot study was undertaken to assess the efficacy of mifepristone and misoprostol, both administered vaginally. The ultimate goal is to investigate alternative means of reducing the time interval between the two treatments involved. The efficacy of the early medical abortion regimen utilizing mifepristone and misoprostol is beyond doubt. The regimen usually involves administering misoprostol 36 h following oral administration of mifepristone. The interval between the two treatment components might affect a woman's choice of the medical method. METHODS: Eighteen women undergoing abortion for nonmedical reasons were recruited. RESULTS: Seven women required further intervention to achieve complete abortion. Median induction-to-abortion interval was 7.66 h in the 11 women with complete abortion. CONCLUSION: The complete abortion rate of 61% in this study was lower than that with the standard medical regimen. PMID- 16472569 TI - Comparing patients' telephone calls after medical and surgical abortions. AB - OBJECTIVE: The purpose of this study is to monitor the content, timing of and response to telephone calls from medical and surgical abortion patients in order to improve the counseling and nursing care and allay patient's concerns. METHODS: There were 43 calls from a possible 626 surgical patients and 100 calls from a possible 671 medical patients. Calls were considered preventable in 67% of the medical patients and 46.5% of the surgical patients. Women who were having their first abortion were more likely to place preventable calls. CONCLUSIONS: Calls could be reduced by explaining variations in normal bleeding and how to use analgesics and providing this information in diagrammatic form on an information sheet. PMID- 16472570 TI - The relationship between source of sexual information and sexual behavior among female adolescents. AB - INTRODUCTION: The relationship between the source of sexual information and adolescent sexual behavior is not well understood. MATERIALS AND METHODS: As part of a larger study, 892 sexually active female adolescents between 15 and 18 years of age completed a paper-and-pencil survey which assessed discussions about sexual intercourse, contraceptive use and condom use among friends and family members and their own sexual behavior. Multivariate logistic regression analyses were performed to examine associations while controlling for key demographics. RESULTS: Adolescents who had never used condoms were less likely to report having had discussions about sexual intercourse with their friends [adjusted odds ratio (AOR) = 0.2]. Discussions about condom use among friends and family members were associated with condom use in the past 3 months, with adolescents who used condoms at least half of the time being more likely to report hearing discussions relative to adolescents who never used condoms (AORs 2.0-3.8). Furthermore, the number of sexual partners was related to whether the adolescent had heard discussions among her family about condom use; the higher the number of sexual partners, the more likely the adolescent was to have heard discussions. For adolescents reporting 2-3, 4-5 and 6 or more partners (relative to one partner), the AORs were 1.7, 2.3 and 2.7, respectively. DISCUSSION: Adolescents exposed to information about condom use among the primary individuals in their lives are more likely to be engaging in this behavior themselves. PMID- 16472571 TI - Hormonal and ovarian stromal blood supply changes after laparoscopic tubal sterilization: a prospective controlled study. AB - OBJECTIVES: The purpose of this study is to determine if the effects of tubal sterilization (TS) by laparoscopy have any risk of a subsequent significant decrease in ovarian reserve and vascular support within the ovary by means of stromal artery Doppler study, and to compare the results with matched paired controls. DESIGN: Between February 2002 and January 2005, 148 healthy volunteers were enrolled sequentially, 74 undergoing laparoscopic TS (study group) and 74 age-, parity-, body mass index-matched women were recruited as a control group. The main outcome measurements were blood levels of follicle-stimulating hormone (FSH), luteinizing hormone and E2, ovarian volume, number of antral follicles and Doppler study of ovarian stromal artery pulsatile index (PI) and maximum velocity (Vmax) on the third day of the cycle immediately before, and 1 and 12 months after the surgical intervention. RESULTS: There were significant elevations in both serum FSH levels and PI values observed 1 month after TS, compared to the preoperative levels (p < .05), and also when compared to controls. However, there were no significant elevations at 12 months postoperation in both study and control groups. The other outcome measurements did not show any significant differences between the two groups. CONCLUSIONS: Tubal sterilization by laparoscopic electrocoagulation does not cause any decrease in ovarian reserve or ovarian stromal blood supply, except an early postoperative increase in FSH and PI. PMID- 16472572 TI - In vitro determination of the contraceptive spermicidal activity of a composite extract of Achyranthes aspera and Stephania hernandifolia on human semen. AB - OBJECTIVE: To evaluate the effect of a 50% ethanolic extract of the leaf of Stephania hernandifolia and the root of Achyranthes aspera on sperm motility and function in a ratio of 1:3 by weight at different concentrations. RESULTS: Concentration of 0.08 g/mL of the extract affected the motility, and at a concentration of 0.16 g/mL, the sperm motility was reduced to 20% immediately (within 20 s). At a concentration of 0.32 g/mL, this composite extract showed the most promising results by complete sperm immobilization within 2 min after the application of the extract. The effects were spermicidal but not spermiostatic as sperm immobilization effect was found to be irreversible. Sperm viability was decreased significantly and was found to be nonviable after 30 min when treated with the composite extract at a concentration of 0.32 g/mL. The hypo-osmotic swelling of these sperm was reduced significantly at this highest concentration, indicating that the crude extract may probably cause injury to the sperm plasma membrane. A low concentration of 0.04 g/mL is ineffective. CONCLUSION: The findings indicate that this composite plant extract possesses potential contraceptive spermicidal activity in vitro. PMID- 16472573 TI - Evaluation of single intratesticular injection of calcium chloride for nonsurgical sterilization in adult albino rats. AB - OBJECTIVE: To investigate a method of chemical sterilization and its efficacy in adult albino rats. METHOD: Evaluation was conducted 3 weeks after a single bilateral intratesticular injection of calcium chloride (CaCl2) at the dose of 5, 10, 15 or 20 mg per testis per 100 g body weight. RESULTS: The significant graded diminution in relative sex organ weights, testicular androgenic enzymes like Delta5,3beta-hydroxysteroid dehydrogenase (Delta5,3beta-HSD) and 17beta hydroxysteroid dehydrogenase (17beta-HSD), glutathione peroxidase (GPx), glutathione S-transferase (GST) and superoxide dismutase (SOD) activities, testicular content of reduced glutathione (GSH), plasma and intratesticular concentrations of testosterone, epididymal sperm count as well as significant elevation in plasma concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH) and testicular content of malondialdehyde (MDA) were noted in all the treated groups with respect to vehicle control. There was no chronic general stress in experimental animals as indicated by insignificant changes in plasma concentrations of corticosterone, prolactin, total protein, blood urea nitrogen and fasting blood sugar level. Dose-dependent responses on testicular histopathology were recorded by noting multinucleated giant cells in seminiferous tubules, derangement of tubular architecture along with infiltration of leucocytes and appearance of fibrous tissue throughout the testicular sections. The fertility efficacy of the 10, 15 or 20 mg CaCl2-treated males was nil, proven after mating with fertile, virgin healthy females, as there were no implantation sites in each uterine horns noted by laparotomy. CONCLUSION: Intratesticular CaCl2 injection at a specific dose might serve as a way of sterilization and may be considered as an alternative to surgical castration in male animals. PMID- 16472574 TI - Evaluation of the potential antifertility effect of fenugreek seeds in male and female rabbits. AB - OBJECTIVE: The objective of this study was to evaluate the potential antifertility activity of feeding diets containing 30% fenugreek seeds to male and female white New Zealand rabbits. RESULTS: The data presented in this study clearly demonstrate an antifertility effect of fenugreek seeds in the female rabbits and more of a toxicity effect in the male rabbits. In males, testis weight was reduced, with evident damage to the seminiferous tubules and interstitial tissues as shown by the histopathology of testis tissue sections. In addition, the plasma concentration of the androgen hormone and sperm concentrations were halved in the treated animals. In the case of the females, there was evidence of a significant reduction of developing fetuses as observed by reductions of both fetal and placental weights at 20 days of gestation and of the litter size. This was further supported histopathologically by the observed proliferative changes of the endometrial glands. The circulating plasma progesterone concentrations at 10 and 20 days of gestation significantly increased with no significant effect on the prebreeding estrogen concentrations in the treated animals. PMID- 16472575 TI - Postcoital ingestion of the aqueous extract of Erythrina falcata Benth prevents pregnancy in the mouse. AB - AIM: We examined whether the aqueous extract of Erythrina falcata, reputed to be a contraceptive in Peruvian folklore, could prevent pregnancy in the mouse. METHODS: Female mice on Day 1 of pregnancy were given aqueous extract of E. falcata or tap water (control) orally for 4 days. On Day 4 of pregnancy, animals were killed and the embryos were flushed from oviducts and uterus to examine their developmental stage, cell number, mitotic index and micronuclei frequency. Other mice were killed on Day 12 of pregnancy to determine the number of implantation sites. RESULTS: Ingestion of E. falcata diminished the percentage of embryos that progressed to blastocyst stage, reduced the cell number and mitotic index, and increased the micronuclei frequency of early embryos. The number of implantation sites was also reduced in females treated with E. falcata. CONCLUSION: The aqueous extract of E. falcata, ingested during early pregnancy, disturbs preimplantation embryo development and implantation in the mouse. These results provide the first experimental evidence of the contraceptive properties of the aqueous extract of E. falcata. PMID- 16472576 TI - Skin venous thromboembolism by combined oral contraceptive in a woman with acquired angioedema and Factor V Leiden mutation. AB - OBJECTIVE: The purpose of this study is to report a case of acquired angioedema and Factor V Leiden mutation in a woman who had started taking an oral contraceptive (OC), and to review the literature. RESULTS: A 44-year-old nonobese and nonsmoking woman was admitted to our hospital because of pain and progressive erythematous lesion on her left groin. The patient had had a 6-week history of combined OC use. There was no family history of angioedema. With a diagnosis of necrotizing fasciitis, surgical debridement of all necrotic tissue was considered with fasciocutaneous flap. Histopathological analysis showed diffusely fresh bleeding areas with diffused subcutaneous necrosis. The culture of the necrotic tissue was negative. Postoperatively, she restarted combined OC, and then the painful petechial and rapidly disseminated ecchymotic lesions were seen. Oral contraceptive was stopped immediately. A venous Doppler ultrasonography of the lower extremities showed subacute thrombosis in the left popliteal vein. Complement C3 level was normal, but amounts of C4, C1 esterase inhibitor level and activity were low. Factor V Leiden was positive for the heterozygous presence of Factor V gene. CONCLUSIONS: Women with angioedema and Factor V Leiden should be counseled against using hormonal contraceptives containing estrogen, and because they may interact in a synergistic manner in the pathogenesis of cutaneous lesions and/or venous thrombosis, appropriate individualized therapy should be defined. Further studies are needed. PMID- 16472577 TI - One-year gross cumulative discontinuation rates are not the rates published by Wildemeersch et al. PMID- 16472583 TI - Living with rejection--lessons for us all. PMID- 16472584 TI - Image of the month. PMID- 16472585 TI - Association of the predominant symptom with clinical characteristics and pathophysiological mechanisms in functional dyspepsia. AB - BACKGROUND & AIMS: Functional dyspepsia (FD) is considered a heterogeneous disorder with different pathophysiological mechanisms contributing to the symptom pattern. The Rome II committee proposed that subdividing patients with FD into groups with predominant pain versus discomfort might identify subgroups with homogeneous pathophysiological and clinical properties. The aim of this study was to analyze the relationship of predominant pain or discomfort with pathophysiological mechanisms and to evaluate whether considering individual predominant symptoms yields better results. METHODS: Consecutive FD patients (n = 720; 489 women; mean age, 41.3 +/- 0.6 years) filled out a dyspepsia questionnaire and identified a single most bothersome symptom. We analyzed the association of this predominant symptom with demographic, clinical, and pathophysiological features (Helicobacter pylori status, gastric emptying in 592 patients, and gastric sensitivity and accommodation testing in 332 patients). RESULTS: According to Rome II criteria, 22% were pain predominant and 78% discomfort predominant. Patients with predominant pain had a higher prevalence of hypersensitivity (44% vs 25%) and delayed gastric emptying was observed less frequently in these patients (16% vs 26%), but there was major overlap. Detailed analysis showed that any of 8 dyspeptic symptoms could be predominant. Predominant early satiety or vomiting was associated with significantly higher prevalences of weight loss (89% and 75%, respectively) and of acute onset (61% and 60%, respectively). Impaired accommodation was found in 79% of patients with predominant early satiety. The highest prevalence of delayed emptying was found in predominant fullness (38%) and of hypersensitivity in predominant pain (44%). CONCLUSIONS: Subdividing FD patient groups according to the predominant symptom does not reliably identify subgroups with a homogeneous underlying pathophysiological mechanism. PMID- 16472586 TI - Hypothalamic-pituitary-gut axis dysregulation in irritable bowel syndrome: plasma cytokines as a potential biomarker? AB - BACKGROUND & AIMS: Irritable bowel syndrome (IBS) is a functional disorder with an etiology that has been linked to both psychological stress and infection. The primary aim of this study was to examine the hypothalamic-pituitary-adrenal axis in patients with IBS and to relate such response to plasma cytokine profiles. METHODS: A total of 151 subjects, 76 patients and 75 controls, were recruited. The patients with IBS were diagnosed according to Rome II criteria. Forty-nine patients and 48 matched controls had cytokine levels measured, and a subset of 21 patients and 21 controls also underwent a corticotropin-releasing hormone (CRH) stimulation test with plasma levels of adrenocorticotropic hormone (ACTH) and cortisol measured. The remaining 27 patients and 27 controls underwent a dexamethasone (1 mg) challenge. RESULTS: Cortisol and the proinflammatory cytokines interleukin (IL)-6 (together with its soluble receptor) and IL-8 were elevated in all IBS subgroups (diarrhea predominant, constipated, and alternators), although the elevation was most marked in the constipated subgroup. There was no alteration in the anti-inflammatory cytokine IL-10. Following CRH infusion, an exaggerated release of both ACTH and cortisol was observed in patients with IBS. There was a significant correlation between the ACTH response (deltaACTH) and the IL-6 levels. A similar relationship existed between the deltaACTH/deltacortisol ratio and the IL-6 levels. Dexamethasone suppression of cortisol was similar in patients and controls. CONCLUSIONS: IBS is characterized by an overactivation of the hypothalamic-pituitary-adrenal axis and a proinflammatory cytokine increase. PMID- 16472587 TI - Heterozygous mutations in PMS2 cause hereditary nonpolyposis colorectal carcinoma (Lynch syndrome). AB - BACKGROUND & AIMS: The role of the mismatch repair gene PMS2 in hereditary nonpolyposis colorectal carcinoma (HNPCC) is not fully clarified. To date, only 7 different heterozygous truncating PMS2 mutations have been reported in HNPCC suspected families. Our aim was to further assess the role of PMS2 in HNPCC. METHODS: We performed Southern blot analysis in 112 patients from MLH1-, MSH2-, and MSH6-negative HNPCC-like families. A subgroup (n = 38) of these patients was analyzed by denaturing gradient gel electrophoresis (DGGE). In a second study group consisting of 775 index patients with familial colorectal cancer, we performed immunohistochemistry using antibodies against MLH1, MSH2, MSH6, and PMS2 proteins. In 8 of 775 tumors, only loss of PMS2 expression was found. In these cases, we performed Southern blot analysis and DGGE. Segregation analysis was performed in the families with a (possibly) deleterious mutation. RESULTS: Seven novel mutations were identified: 4 genomic rearrangements and 3 truncating point mutations. Three of these 7 families fulfill the Amsterdam II criteria. The pattern of inheritance is autosomal dominant with a milder phenotype compared with families with pathogenic MLH1 or MSH2 mutations. Microsatellite instability and immunohistochemical analysis performed in HNPCC-related tumors from proven carriers showed a microsatellite instability high phenotype and loss of PMS2 protein expression in all tumors. CONCLUSIONS: We show that heterozygous truncating mutations in PMS2 do play a role in a small subset of HNPCC-like families. PMS2 mutation analysis is indicated in patients diagnosed with a colorectal tumor with absent staining for the PMS2 protein. PMID- 16472588 TI - Human anti-tumor necrosis factor monoclonal antibody (adalimumab) in Crohn's disease: the CLASSIC-I trial. AB - BACKGROUND & AIMS: Tumor necrosis factor blockade has been shown to be an effective treatment strategy in Crohn's disease (CD). Adalimumab is a human immunoglobulin G1 (IgG(1)) monoclonal antibody targeting tumor necrosis factor (TNF). A randomized, double-blind, placebo-controlled, dose-ranging trial was performed to evaluate the efficacy of adalimumab induction therapy in patients with CD. METHODS: A total of 299 patients with moderate to severe CD naive to anti-TNF therapy were randomized to receive subcutaneous injections at weeks 0 and 2 with adalimumab 40 mg/20 mg, 80 mg/40 mg, or 160 mg/80 mg or placebo. The primary endpoint was demonstration of a significant difference in the rates of remission at week 4 (defined as a Crohn's Disease Activity Index score <150 points) among the 80 mg/40 mg, 160 mg/80 mg, and placebo groups. RESULTS: The rates of remission at week 4 in the adalimumab 40 mg/20 mg, 80 mg/40 mg, and 160 mg/80 mg groups were 18% (P = .36), 24% (P = .06), and 36% (P = .001), respectively, and 12% in the placebo group. Adverse events occurred at similar frequencies in all 4 treatment groups except injection site reactions, which were more common in adalimumab-treated patients. CONCLUSIONS: Adalimumab was superior to placebo for induction of remission in patients with moderate to severe Crohn's disease naive to anti-TNF therapy. The optimal induction dosing regimen for adalimumab in this study was 160 mg at week 0 followed by 80 mg at week 2. Adalimumab was well tolerated. PMID- 16472589 TI - Intermittent spatial separation of diaphragm and lower esophageal sphincter favors acidic and weakly acidic reflux. AB - BACKGROUND & AIMS: In small hiatal hernias, the size of the hernia is variable. Intermittent complete reduction can be observed with high-resolution manometry as a transition from a double-peak (hernia) to a single-peak (reduced) high-pressure zone. The aim of this study was to investigate whether intermittent separation of the diaphragm and lower esophageal sphincter (LES) favors the occurrence of gastroesophageal reflux. METHODS: In 16 patients with a small hiatal hernia (3 cm), prolonged high-resolution manometry was performed. Acid and weakly acidic reflux episodes were detected with pH-impedance monitoring. RESULTS: The single pressure peak profile (reduced hernia) was present for 814 minutes (56.5% of total time), and the double peak profile (unreduced hernia) was present for 626 minutes (43.5% of total time). In all patients, both pressure profiles were observed. The transition rate between the 2 profiles was 7.5 +/- 0.9 per hour. More reflux occurred when the LES and diaphragm were separated versus the reduced hernia state (23.1 +/- 5.1 vs 12.2 +/- 2.4 episodes per hour, respectively; P < .05). The proportions of acidic reflux episodes during the single and double pressure peaks were similar (70% and 67%, respectively). In the two-pressure-zone state, there was an increase in all reflux mechanisms except transient LES relaxation. CONCLUSIONS: In patients with a small hiatal hernia, intermittent reduction of the hernia occurs frequently. Spatial separation of the diaphragm and LES in the nonreduced state results in a 2-fold increase in acidic and weakly acidic reflux due to mechanisms other than transient LES relaxation. PMID- 16472590 TI - The pregnane X receptor locus is associated with susceptibility to inflammatory bowel disease. AB - BACKGROUND & AIMS: The pregnane X receptor (PXR) regulates an array of genes involved in the response to xenobiotics. Evidence from several studies suggests that xenobiotic metabolism may play a role in inflammatory bowel disease (IBD) and that low levels of PXR may be associated with disease expression. The aim of this study was to investigate the association of functional polymorphisms of the PXR encoding gene (NR1I2) with disease in IBD populations. METHODS: This was a case-control study examining 8 NR1I2 single nucleotide polymorphisms (SNPs) previously associated with altered activity of PXR-regulated genes in an Irish cohort including 422 patients with IBD and 350 ethnically matched controls. RESULTS: We showed significant associations of NR1I2 with IBD, Crohn's disease (CD), and ulcerative colitis (UC) groups compared with a control population for SNPs -23585 (IBD: P = .000008; odds ratio [OR], 1.62; 95% confidence interval [CI], 1.31-2.00) and -24381 (IBD: P = .0002; OR, 1.50; 95% CI, 1.21-1.84). SNPs 7635 (P = .0008) and 8055 (P = .007) were found to be associated with IBD and CD but not UC. Risk of IBD is strongly correlated to genotype at these sites, especially for the -25385CC genotype (P = .00001; OR, 2.92; 95% CI, 1.87-4.66). We also show specific correlations of IBD phenotype with genotypes and haplotypes in the patient group. CONCLUSIONS: These results show that genetic variation in the PXR encoding gene, which has been associated with altered activity of PXR, is strongly associated with susceptibility to IBD, CD, and UC. PMID- 16472591 TI - Involvement of the anion exchanger SLC26A6 in prostaglandin E2- but not forskolin stimulated duodenal HCO3- secretion. AB - BACKGROUND & AIMS: SLC26A6 is a recently identified apical Cl(-)/HCO(3)(-) exchanger with strong expression in murine duodenum. The present study was designed to examine the role of SLC26A6 in prostaglandin E(2) (PGE(2))-, forskolin-, and carbachol-induced duodenal HCO(3)(-) secretion. METHODS: Murine duodenal mucosal HCO(3)(-) secretion was examined in vitro in Ussing chambers and mucosal SLC26A6 expression levels were analyzed by semiquantitative reverse transcription polymerase chain reaction. RESULTS: Basal HCO(3)(-) secretion was diminished by 20%, PGE(2)-stimulated HCO(3)(-) secretory response by 59%, and carbachol-stimulated response was reduced by 35% in SLC26A6-/- compared with +/+ duodenal mucosa, whereas the forskolin-stimulated HCO(3)(-) secretory response was not different. In Cl(-)-free solutions, PGE(2)- and carbachol-stimulated HCO(3)(-) secretion was reduced by 81% and 44%, respectively, whereas forskolin stimulated HCO(3)(-) secretion was not altered significantly. PGE(2) and carbachol, but not forskolin, were able to elicit a Cl(-)-dependent HCO(3)(-) secretory response in the absence of short-circuit current changes in cystic fibrosis transmembrane conductance regulator knockout mice. CONCLUSIONS: In murine duodenum, PGE(2)-mediated HCO(3)(-) secretion is strongly SLC26A6 dependent and cystic fibrosis transmembrane conductance regulator independent, whereas forskolin-stimulated HCO(3)(-) secretion is completely SLC26A6 independent and cystic fibrosis transmembrane conductance regulator dependent. Carbachol-induced secretion is less pronounced, but occurs via both transport pathways. This suggests that PGE(2) and forskolin activate distinct HCO(3)(-) transport pathways in the murine duodenum. PMID- 16472593 TI - Suppressor of cytokine signaling-1 regulates inflammatory bowel disease in which both IFNgamma and IL-4 are involved. AB - BACKGROUND & AIMS: The suppressor of cytokine signaling-1 (SOCS1) is a potent negative regulator of various cytokines and it has been implicated in the regulation of immune responses. However, the role of SOCS1 in inflammatory bowel diseases (IBDs) has not been clarified. To determine the role of SOCS1 in colitis, we generated SOCS1/T-cell receptor alpha (TCRalpha) double knockout (DKO) mice. METHODS: The depletion of interferon gamma (IFNgamma) and IL-4 was achieved by crossing the DKO mice with IFNgamma knockout (KO) mice and by the administration of anti-IL-4 antibody, respectively. The activation of cytokine induced transcription factors was determined by Western blotting with phosphorylation-specific antibodies, and the induction of inflammatory factors was measured by reverse-transcription polymerase chain reaction. RESULTS: Much more severe colitis developed in 100% of the DKO mice within 9 weeks of age than in TCRalpha-KO mice. Although the proportion and the activation status of CD4(+) TCRalpha(-)beta(+) T cells in DKO mice were similar to those in TCRalpha-KO mice, signal transducer and activator of transcription 1, nuclear factor kappaB, and their target genes were hyperactivated in infiltrated mononuclear cells and colonic epithelial cells in DKO mice. Cytokine-depletion experiments showed that exacerbated colitis in the DKO mice was dependent on both IFNgamma and IL-4. SOCS1-deficient cells were hypersensitive to IFNgamma, IL-4, and lipopolysaccharides, depending on the target genes. CONCLUSIONS: SOCS1 plays an important role in preventing murine colitis by restricting the cytokine signals. SOCS1/TCRalpha DKO mice could be a useful model for investigating human IBD. PMID- 16472592 TI - Bile acids induce ileal damage during experimental necrotizing enterocolitis. AB - BACKGROUND & AIMS: Necrotizing enterocolitis (NEC) is the most common gastrointestinal emergency of premature infants. While the effect of bile acids (BAs) on intestinal mucosal injury is known, we investigated the contribution of BAs during the development of NEC in neonatal rats. METHODS: Premature rats were fed with cow's milk-based formula and subjected to asphyxia and cold stress to develop NEC. Jejunal and ileal luminal BAs, portal blood BAs, and messenger RNA and protein for the apical sodium-dependent bile acid transporter, the ileal bile acid binding protein, and the heteromeric organic solute transporter (Ostalpha/Ostbeta)were evaluated. RESULTS: Ileal luminal BAs levels were increased significantly during disease development and the removal of ileal BAs significantly decreased the incidence and severity of disease. Furthermore, when NEC was reduced via treatment with epidermal growth factor (EGF), BA levels were reduced significantly. Jejunal luminal BA levels were similar between animals with NEC and controls, but portal/ileal luminal BA ratios were decreased significantly in animals with NEC. The apical sodium-dependent bile acid transporter was up-regulated at the site of injury in animals with NEC and decreased after EGF treatment; however, the ileal bile acid binding protein was up-regulated only in the NEC and EGF group. Ostalpha/Ostbeta expression was low in all groups, and only slightly increased in the NEC group. CONCLUSIONS: These data strongly suggest that BAs play a role in the development of ileal damage in experimental NEC and that alterations in BA transport in the neonatal ileum may contribute to disease development. PMID- 16472594 TI - A functional role of flip in conferring resistance of Crohn's disease lamina propria lymphocytes to FAS-mediated apoptosis. AB - BACKGROUND & AIMS: There is evidence that, in Crohn's disease (CD), lamina propria T lymphocytes (LPLs) are resistant to FAS-mediated apoptosis and that this defect contributes to the mucosal T-cell accumulation. In this study we examined the functional role of Flip, a Flice inhibitor protein, in the resistance of CD LPL to FAS-mediated apoptosis. METHODS: Biopsy specimens and LPLs were taken from CD and ulcerative colitis (UC) patients and normal controls and analyzed for Flip by Western blotting. We also examined whether inhibition of Flip by antisense oligonucleotide restored the susceptibility of CD LPLs to FAS induced apoptosis. LPL apoptosis was assessed by flow cytometry. RESULTS: After FAS stimulation, the rate of apoptosis of CD3+ LPLs was higher in normal controls and patients with UC than in patients with CD. Enhanced expression of both long and short Flip isoforms was seen in biopsy specimens and purified CD3+ and CD45RO+ LPLs of CD patients in comparison with UC patients and normal controls. No increase in Flip was documented in untreated celiac disease mucosa, thus suggesting the possibility that induction of Flip in the gut does not simply rely on the ongoing inflammation. Finally, we showed that inhibition of Flip by antisense oligonucleotide reverted the resistance of CD LPLs to FAS-induced apoptosis. CONCLUSIONS: Data suggest a role for Flip in the resistance of CD LPLs to FAS-mediated apoptosis. PMID- 16472595 TI - Chitinase 3-like-1 exacerbates intestinal inflammation by enhancing bacterial adhesion and invasion in colonic epithelial cells. AB - BACKGROUND & AIMS: Dysregulated host/microbial interactions appear to play a central role in the development of inflammatory bowel disease (IBD). However, molecular events leading to the dysregulation have not yet been defined fully. Studies were designed to characterize a key molecule that is involved in the dysregulation. METHODS: Colonic mucosal RNA from C57BL/6 mice on days 4 and 8 with administration of 4% dextran sulfate sodium for 5 days were subjected to DNA microarray analysis. Chitinase 3-like-1 (CHI3L1) messenger RNA and protein expressions were examined by reverse-transcription polymerase chain reaction and immunohistochemistry. A gentamicin protection assay of Salmonella typhimurium was performed using epithelial cell lines that are engineered genetically to overexpress or lack mouse CHI3L1. To examine the functional role of CHI3L1 in vivo, anti-CHI3L1 antibody was administered into the dextran sulfate sodium colitis model. RESULTS: Microarray analysis identified that CHI3L1 is up regulated specifically in inflamed mucosa. The expression of CHI3L1 protein clearly was detectable in lamina propria and colonic epithelial cells (CECs) in several murine colitis models and ulcerative colitis and Crohn's disease patients but absent in normal controls. The gentamicin protection assays using intracellular bacteria showed that CHI3L1 is required for the enhancement of adhesion and internalization of these bacteria in CEC. In vivo neutralization experiments showed that CHI3L1 contributes to the facilitation of bacterial invasion into the intestinal mucosa and the development of acute colitis. CONCLUSIONS: CHI3L1 plays a pathogenic role in colitis, presumably by enhancing the adhesion and invasion of bacteria on/into CEC. Inhibition of CHI3L1 activity would be a novel therapeutic approach for IBD. PMID- 16472596 TI - Epidermal growth factor receptor signaling regulates Bax and Bcl-w expression and apoptotic responses during intestinal adaptation in mice. AB - BACKGROUND & AIMS: Normal intestinal adaptation to massive small-bowel resection requires intact epidermal growth factor receptor signaling and consists of increased enterocyte proliferation and apoptosis. Although emphasis has been placed on understanding the regulation of proliferation, few studies have evaluated the mechanism and contribution of apoptosis to the adaptation response. We sought to test the hypothesis that epidermal growth factor receptor signaling regulates specific Bcl-2 family members (Bax and Bcl-w) to direct apoptosis and adaptation after massive small-bowel resection. METHODS: Laser capture microdissection microscopy permitted measurement of Bax and Bcl-w messenger RNA expression in crypt and villus enterocytes in control conditions and under epidermal growth factor receptor-inhibited (waved-2 mice) or stimulated (epidermal growth factor transgenic mice) conditions after a 50% small-bowel resection or sham operation. Resection-induced adaptation was then studied in Bax null and Bcl-w-null mice under control circumstances and after epidermal growth factor receptor stimulation. RESULTS: When compared with Bcl-w, the most significant expression changes were observed with Bax and took place within crypt enterocytes. Epidermal growth factor receptor stimulation resulted in a decreased ratio of Bax to Bcl-w expression and decreased rates of apoptosis. Bax-null mice had no apoptosis response to small-bowel resection and displayed an amplified adaptation response to the administration of epidermal growth factor. Bcl-w-null mice had poor survival and impaired adaptation to small-bowel resection, an effect that was rescued by crossbreeding these mice with epidermal growth factor transgenic mice. CONCLUSIONS: The crypt expression of Bax and Bcl-w is influenced by epidermal growth factor receptor signaling and is key for the regulation of apoptosis. Epidermal growth factor receptor stimulation, coupled with apoptosis inhibition, may provide a novel strategy to amplify adaptation responses in patients after massive intestinal loss. PMID- 16472597 TI - CD48 controls T-cell and antigen-presenting cell functions in experimental colitis. AB - BACKGROUND & AIMS: The cell-surface receptor CD48 is a lipid-anchored protein expressed on all antigen-presenting cells and T cells. CD2 and 2B4 are known ligands for CD48, which themselves are expressed on the surface of hematopoietic cells. Here we examine the effect of CD48 in the development of chronic experimental colitis and how CD48 affects adaptive and innate immune functions. METHODS: The role of CD48 in experimental colitis was first assessed by transferring CD4(+)CD45RB(hi) cells isolated from either wild-type or CD48(-/-) mice into either Rag-2(-/-) or CD48(-/-) x Rag-2(-/-) mice. Development of chronic colitis in these adoptively transferred mice was assessed by disease activity index, histology, and production of interferon-gamma in mesenteric lymph nodes. Relevant functions of CD48(-/-)CD4(+) T cells and CD48(-/-) macrophages were examined using in vitro assays. In a second set of experiments, the efficacy of anti-CD48 in prevention or treatment of chronic colitis was determined. RESULTS: CD48(-/-)CD4(+) cells induced colitis when transferred into Rag-2(-/-) mice, but not when introduced into CD48(-/-) x Rag-2(-/-) recipients. However, both recipient mouse strains developed colitis upon adoptive transfer of wild type CD4(+) cells. Consistent with a CD4(+) T-cell defect was the observation that in vitro proliferation of CD48(-/-)CD4(+) T cells was impaired upon stimulation with CD48(-/-) macrophages. In vitro evidence for a modest macrophage functional defect was apparent because CD48(-/-) macrophages produced less tumor necrosis factor alpha and interleukin 12 than wild-type cells upon stimulation with lipopolysaccharide. Peritoneal macrophages also showed a defect in clearance of gram-negative bacteria in vitro. Treatment of the CD4(+)CD45RB(hi)-->Rag-2(-/ ) mice or the wild-type BM-->tg26 mice with anti-CD48 (HM48-1) ameliorated development of colitis, even after its induction. CONCLUSIONS: Both CD48 dependent activation of macrophages and CD48-controlled activation of T cells contribute to maintaining the inflammatory response. Consequently, T cell-induced experimental colitis is ameliorated only when CD48 is absent from both T cells and antigen-presenting cells. Because anti-CD48 interferes with these processes, anti-human CD48 antibody treatment may represent a novel therapy for inflammatory bowel disease patients. PMID- 16472598 TI - Natural killer cells ameliorate liver fibrosis by killing activated stellate cells in NKG2D-dependent and tumor necrosis factor-related apoptosis-inducing ligand-dependent manners. AB - BACKGROUND & AIMS: Viral hepatitis infection, which is a major cause of liver fibrosis, is associated with activation of innate immunity. However, the role of innate immunity in liver fibrosis remains obscure. METHODS: Liver fibrosis was induced either by feeding mice with the 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) diet or by injecting them with carbon tetrachloride. The Toll-like receptor 3 ligand, polyinosinic-polycytidylic acid, was used to activate innate immunity cells and mediators, including natural killer cells and interferon gamma. RESULTS: In the mouse model of DDC-induced liver fibrosis, natural killer cell activation by polyinosinic-polycytidylic acid induced cell death to activated hepatic stellate cells and attenuated the severity of liver fibrosis. Polyinosinic-polycytidylic acid treatment also ameliorated liver fibrosis induced by carbon tetrachloride. The observed protective effect of polyinosinic polycytidylic acid on liver fibrosis was diminished through either depletion of natural killer cells or by disruption of the interferon gamma gene. Expression of retinoic acid early inducible 1, the NKG2D ligand, was undetectable on quiescent hepatic stellate cells, whereas high levels were found on activated hepatic stellate cells, which correlated with the resistance and susceptibility of quiescent hepatic stellate cells and activated hepatic stellate cells to natural killer cell lysis, respectively. Moreover, treatment with polyinosinic polycytidylic acid or interferon gamma enhanced the cytotoxicity of natural killer cells against activated hepatic stellate cells and increased the expression of NKG2D and tumor necrosis factor-related apoptosis-inducing ligand on liver natural killer cells. Blocking NKG2D or tumor necrosis factor-related apoptosis-inducing ligand with neutralizing antibodies markedly diminished the cytotoxicity of polyinosinic-polycytidylic acid-activated natural killer cells against activated hepatic stellate cells. CONCLUSIONS: Our findings suggest that natural killer cells kill activated hepatic stellate cells via retinoic acid early inducible 1/NKG2D-dependent and tumor necrosis factor-related apoptosis inducing ligand-dependent mechanisms, thereby ameliorating liver fibrosis. PMID- 16472600 TI - 24-norUrsodeoxycholic acid is superior to ursodeoxycholic acid in the treatment of sclerosing cholangitis in Mdr2 (Abcb4) knockout mice. AB - BACKGROUND & AIMS: Current therapy for primary sclerosing cholangitis is of limited efficacy. Multidrug resistance gene 2 knockout mice (Mdr2(-/-)) represent a well-characterized model for sclerosing cholangitis. Experiments were performed to test in such mice the therapeutic effects of 24-norUrsodeoxycholic acid, a C(23) homologue of ursodeoxycholic acid with 1 fewer methylene group in its side chain. METHODS: Mdr2(-/-) mice were fed a diet containing 24-norUrsodeoxycholic acid (0.5% wt/wt) or ursodeoxycholic acid (0.5% wt/wt) as a clinical comparator for 4 weeks; controls received standard chow. Effects on serum liver tests, liver histology, markers of inflammation and fibrosis, and bile acid transport and metabolism were compared. 24-norUrsodeoxycholic acid metabolism was studied in serum, liver, bile, and urine. RESULTS: 24-norUrsodeoxycholic acid markedly improved liver tests and liver histology and significantly reduced hydroxyproline content and the number of infiltrating neutrophils and proliferating hepatocytes and cholangiocytes. 24-norUrsodeoxycholic acid underwent extensive phase I/II metabolism (hydroxylation, sulfation, and glucuronidation), thereby increasing the hydrophilicity of biliary bile acid secretion. There was a coordinated induction of bile acid detoxifying enzymes (Cyp2b10, Cyp3a11, and Sult2a1) and efflux pumps (Mrp3 and Mrp4). Ursodeoxycholic acid, in contrast, increased alanine transaminase and alkaline phosphatase levels, had no significant effects on hydroxyproline content, and induced biliary transporters and detoxification enzymes to a much smaller extent than 24-norUrsodeoxycholic acid. CONCLUSIONS: 24 norUrsodeoxycholic acid ameliorates sclerosing cholangitis in Mdr2(-/-) mice. Its therapeutic mechanisms involve (1) increasing the hydrophilicity of biliary bile acids, (2) stimulating bile flow with flushing of injured bile ducts, and (3) inducing detoxification and elimination routes for bile acids. PMID- 16472599 TI - Vaccination with protein-transduced dendritic cells elicits a sustained response to hepatitis C viral antigens. AB - BACKGROUND & AIMS: Professional antigen-presenting dendritic cells are capable of eliciting a vigorous antiviral response in naive T cells. The administration of antigen-loaded dendritic cells offers a potential approach to induce high-level immunity against hepatitis C virus. METHODS: The dendritic cell population in mice was expanded in vivo by hydrodynamic delivery of naked DNA that encoded the secreted form of human fms-like tyrosine kinase 3 ligand. The CD11c-enriched dendritic cell population obtained from the spleen was transduced in vitro with recombinant hepatitis C virus core and nonstructural 5 proteins by using macromolecular-based protein delivery. Vaccine efficacy was assessed with a cytotoxic T-lymphocyte assay, cytokine enzyme-linked immunosorbent assays, and intracellular cytokine staining in vitro and by a tumor challenge model in vivo. RESULTS: Relative to mice inoculated with nontransduced dendritic cells, splenocytes derived from mice immunized with either hepatitis C virus core transduced or nonstructural 5-transduced dendritic cells showed 3- to 5-fold greater antigen-specific cytotoxic T lymphocyte activity. The CD4(+) T cells obtained from mice immunized with nonstructural 5-transduced dendritic cells produced interferon gamma, but not interleukin 4, when stimulated with nonstructural 5. In contrast, T cells derived from mice immunized with hepatitis C virus core-transduced dendritic cells produced neither interferon gamma nor interleukin 4 when stimulated with core protein. Mice vaccinated with nonstructural 5-transduced dendritic cells, but not a nonstructural 5-expressing plasmid, showed a sustained antiviral response to nonstructural 5 as evidenced by reduced growth of nonstructural 5-expressing tumor cells inoculated 10 weeks after vaccination. CONCLUSIONS: These findings suggest that vaccination with protein-transduced dendritic cells may constitute an important antiviral strategy for hepatitis C virus. PMID- 16472601 TI - The hepatitis C envelope 2 protein inhibits LFA-1-transduced protein kinase C signaling for T-lymphocyte migration. AB - BACKGROUND & AIMS: The ability of viruses to escape the host immune response represents a globally important problem related to a wide variety of pathogens. Hepatitis C is one of the major causes of liver disease worldwide. Clearance rates of this virus are low, and this condition normally involves a chronic inflammatory process. This raises a possibility that the virus may have developed mechanisms enabling it to evade T-cell-mediated immune surveillance. The aim of this study was to investigate the effect of the hepatitis C envelope protein E2 on LFA-1-stimulated T-cell migration and macrophage inflammatory protein (MIP 1alpha, MIP-1beta) secretion. METHODS: T cells were stimulated through the leukocyte function-associated molecule-1 (LFA-1) receptor by incubating with either intracellular adhesion molecule 1 (ICAM-1)-Fc fusion protein or anti-LFA-1 immobilized on 8-well chamber slides. Subcellular localization of protein kinase C (PKC)-beta, CD81, and LFA-1 was determined by immunofluorescence analysis. Lipid raft formation was assessed using the Cellomics Kineticscan reader. MIP 1alpha and MIP-1beta levels were detected by enzyme-linked immunosorbent assay. RESULTS: We report that the hepatitis C envelope protein E2 can dramatically inhibit T-lymphocyte motility and chemokine release induced via LFA-1 integrin ligation. We have demonstrated a novel T-lymphocyte-directed viral inhibitory mechanism involving the PKC-beta enzyme as a definitive intracellular target. E2 CD81 interaction stimulates translocation of PKC-beta to lipid rafts, thereby preventing its association with the centrosome and microtubule cytoskeleton, which is crucial to the process of T-cell migration. CONCLUSIONS: These studies identify a mechanism whereby the hepatitis C virus can evade the host immune response by inhibition of T-cell migration. PMID- 16472602 TI - ATP7B mediates vesicular sequestration of copper: insight into biliary copper excretion. AB - BACKGROUND & AIMS: The Wilson protein (ATP7B) regulates levels of systemic copper by excreting excess copper into bile. It is not clear whether ATP7B translocates excess intrahepatic copper directly across the canalicular membrane or sequesters this copper into exocytic vesicles, which subsequently fuse with canalicular membrane to expel their contents into bile. The aim of this study was to clarify the mechanism underlying ATP7B-mediated copper detoxification by investigating endogenous ATP7B localization in the HepG2 hepatoma cell line and its ability to mediate vesicular sequestration of excess intracellular copper. METHODS: Immunofluorescence microscopy was used to investigate the effect of copper concentration on the localization of endogenous ATP7B in HepG2 cells. Copper accumulation studies to determine whether ATP7B can mediate vesicular sequestration of excess intracellular copper were performed using Chinese hamster ovary cells that exogenously expressed wild-type and mutant ATP7B proteins. RESULTS: In HepG2 cells, elevated copper levels stimulated trafficking of ATP7B to pericanalicular vesicles and not to the canalicular membrane as previously reported. Mutation of an endocytic retrieval signal in ATP7B caused the protein to constitutively localize to vesicles and not to the plasma membrane, suggesting that a vesicular compartment(s) is the final trafficking destination for ATP7B. Expression of wild-type and mutant ATP7B caused Chinese hamster ovary cells to accumulate copper in vesicles, which subsequently undergo exocytosis, releasing copper across the plasma membrane. CONCLUSIONS: This report provides compelling evidence that the primary mechanism of biliary copper excretion involves ATP7B mediated vesicular sequestration of copper rather than direct copper translocation across the canalicular membrane. PMID- 16472603 TI - Cell competition leads to a high level of normal liver reconstitution by transplanted fetal liver stem/progenitor cells. AB - BACKGROUND & AIMS: A critical property of stem cells is their ability to repopulate an organ or tissue under nonselective conditions. The aims of this study were to determine whether we could obtain reproducible, high levels of liver repopulation by transplanted fetal liver stem/progenitor cells in normal adult liver and the mechanism by which liver replacement occurred. METHODS: Wild type (dipeptidyl peptidase IV [DPPIV(+)]) embryonic day (ED) 14 fetal liver cells underwent transplantation into DPPIV(-) mutant F344 rats to follow the fate and differentiation of transplanted cells. To determine the mechanism for repopulation, proliferation and apoptosis of transplanted and host liver cells were also followed. RESULTS: Transplanted ED 14 fetal liver cells proliferated continuously for 6 months, differentiated into mature hepatocytes, and replaced 23.5% of total liver mass. The progeny of transplanted cells were morphologically and functionally indistinguishable from host hepatocytes and expressed unique liver-specific genes commensurate with their location in the hepatic lobule. Repopulation was based on greater proliferative activity of transplanted cells and reduced apoptosis of their progeny compared with host hepatocytes, coupled with increased apoptosis of host hepatocytes immediately adjacent to transplanted cells. This process, referred to as cell-cell competition, has been described previously in Drosophila during wing development. CONCLUSIONS: We show for the first time that cell-cell competition, a developmental paradigm, can be used to replace functional organ tissue in an adult mammalian species under nonselective conditions and may serve as a strategy for tissue reconstitution in a wide variety of metabolic and other disorders involving the liver, as well as other organs. PMID- 16472604 TI - Endothelial progenitor cell transplantation improves the survival following liver injury in mice. AB - BACKGROUND & AIMS: Neovascularization, which is vital to the healing of injured tissues, recently has been found to include both angiogenesis, which involves in mature endothelial cells, and vasculogenesis, involving endothelial progenitor cells. The aim of this study was to clarify the possible roles of endothelial progenitor cells during postnatal liver regeneration. METHODS: To determine how endothelial progenitor cells participate in liver regeneration, human or mouse endothelial progenitor cells were transplanted into the mice with carbon tetrachloride-induced acute liver injury. Survival rate of the mice in endothelial progenitor cell-transplanted and control groups was calculated. Separately, livers removed temporally from both groups were examined. RESULTS: At an early stage, transplanted human endothelial progenitor cells were seen mainly surrounding hepatic central veins where hepatocytes showed extensive necrosis; later, the transplanted cells formed tubular structures. More of these cells were observed along hepatic sinusoids. Transplantation of human or mouse endothelial progenitor cells improved survival of the mice following liver injury (from 28.6% to 85.7%, P < .0005 and from 33.3% to 80.0%, P < .001, respectively), accompanied by greater proliferation of hepatocytes. Human endothelial progenitor cells produced several growth factors, such as hepatocyte growth factor, transforming growth factor-alpha, heparin-binding epidermal growth factor-like growth factor, and vascular endothelial growth factor, and also elicited endogenous growth factors. CONCLUSIONS: Endogenous and exogenous growth factors and direct neovascularization after endothelial progenitor cell transplantation promoted liver regeneration, thus improving survival after liver injury. Transplantation of endothelial progenitor cells could represent a new therapeutic strategy for promoting liver regeneration. PMID- 16472605 TI - The transcription factor hepatocyte nuclear factor-6 controls the development of pancreatic ducts in the mouse. AB - BACKGROUND & AIMS: A number of hereditary polycystic diseases are associated with formation of cysts within the pancreatic ducts. The cysts result from abnormal tubulogenesis, but how normal pancreatic duct development is controlled remains poorly understood. Here, we investigate the transcriptional mechanisms that control pancreatic duct development by addressing the role of the transcription factor hepatocyte nuclear factor (HNF)-6. METHODS: Using immunostaining, we have determined the expression pattern of HNF-6 in pancreatic ducts during mouse development. Hnf6 null mice at various stages of development were studied by immunolocalization methods to assess the morphology, differentiation, and proliferation status of ductal cells. The expression of genes involved in hereditary polycystic diseases was determined by real-time, reverse-transcription polymerase chain reaction (RT-PCR). RESULTS: We show that HNF-6 is expressed in the pancreatic duct epithelium throughout development and that, in the absence of HNF-6, duct morphogenesis is perturbed. Although development of the intercalated ducts is normal, cysts appear within the interlobular and intralobular ducts. This is associated with abnormal development of primary cilia at the apical pole of the duct cells and with reduced expression of a set of genes involved in polycystic diseases, namely those coding for HNF-1beta and for the cilium associated proteins polyductin/fibrocystin and cystin. CONCLUSIONS: We identify HNF-6 as the first transcriptional regulator of pancreatic duct development and reveal the existence of different regulatory mechanisms in distinct duct compartments. HNF-6 controls a network of genes involved in cilium formation and in hereditary polycystic diseases. Finally, HNF-6 deficiency represents a genetically defined model of pancreatic cystic disease. PMID- 16472606 TI - Liver transplantation in a patient with sitosterolemia and cirrhosis. AB - Sitosterolemia (MIM 210250) is a rare genetic disorder caused by disruption of the normal mechanisms that regulate dietary cholesterol absorption and prevent the accumulation of noncholesterol sterols. As a result of this defect, affected individuals accumulate high concentrations of plant sterols in plasma and tissues. They present clinically with tendon or tuberoeruptive xanthomas, premature coronary artery disease, and/or hemolytic anemia. Two genes, ABCG5 and ABCG8, compose the STSL locus, and complete mutation in either, but not both, results in disease. The expression of these genes is confined to the intestine and liver. They are thought to function as sterol efflux pumps. It is not clear which organ, liver or intestine, is of greater importance in maintaining sterol balance with respect to noncholesterol sterols. We report here a case of a patient who presented with "chronic active liver disease" and was found to have sitosterolemia by chance and subsequently underwent orthotopic liver transplantation. Following transplantation, the grossly elevated pretransplantation serum plant sterol levels decreased to values only slightly higher than those of the patient's heterozygous father. This case highlights 2 important features: (1) The liver functions as the predominant organ for maintaining noncholesterol sterol balance (because the intestinal defect was not altered), and (2) a new clinical feature of undiagnosed sitosterolemia may be "idiopathic" liver disease. Because the diagnosis of sitosterolemia is based on specialized plasma analyses, we would propose that some consideration to this diagnosis should be given in appropriate cases. PMID- 16472607 TI - Differential and epigenetic gene expression profiling identifies frequent disruption of the RELN pathway in pancreatic cancers. AB - BACKGROUND & AIMS: Recently described genome-wide approaches robustly detect many candidate genes that are regulated by DNA methylation, but many of these genes do not represent important targets for functional inactivation. Here we used a microarray-based strategy to identify biologically relevant genes associated with epigenetic silencing in pancreatic cancer. METHODS: We compared information from differential gene expression analysis with the transcriptional responses to epigenetic modifiers. RESULTS: Using this approach, we identified 7 novel targets for aberrant methylation in pancreatic cancer. One of the genes identified, RELN (Reelin), a key regulator of neuronal migration, is frequently silenced in pancreatic cancers, as are several of its downstream mediators. Importantly, small interfering RNA-mediated knockdown of RELN in pancreatic cancer cells that retain RELN expression resulted in greatly enhanced cell motility, invasiveness, and colony-forming ability. Increased cell motility was also induced by knockdown of downstream components of the RELN pathway, including ApoER2, VLDLR, and DAB1. Treatment of pancreatic cancer cells with histone deacetylase inhibitors, valproic acid and suberoylanilide hydroxamic acid, restored the expression of RELN and DAB1 and markedly inhibited their migration. CONCLUSIONS: The high prevalence of the silencing of RELN pathway components and its reversal by histone deacetylase inhibitors suggest the importance of this pathway as a diagnostic and therapeutic target for pancreatic cancer. PMID- 16472608 TI - Nonpolypoid (flat and depressed) colorectal neoplasms. PMID- 16472609 TI - New developments in Lynch syndrome (hereditary nonpolyposis colorectal cancer) and mismatch repair gene testing. PMID- 16472610 TI - Drawing a blank in functional dyspepsia? PMID- 16472611 TI - Neuroendocrine and neuroimmune markers in IBS: pathophysiological role or epiphenomenon? PMID- 16472612 TI - Activation-induced cell death of hepatic stellate cells by the innate immune system. PMID- 16472613 TI - The dendritic cell: the immune system's adjuvant--a strategy to develop a HCV vaccine? PMID- 16472614 TI - Azathioprine versus mesalamine in steroid-dependent ulcerative colitis: long awaited results? PMID- 16472616 TI - Where do we go from HEIRS? PMID- 16472618 TI - Searching for the bitter truth: insights into receptors for taste. PMID- 16472619 TI - Combination therapy for chronic hepatitis B: a one-two knockout punch, or a swing and a miss? PMID- 16472621 TI - New frontier in wireless endoscopy: Crohn's disease. PMID- 16472622 TI - Expression of hedgehog pathway components in the adult colon. PMID- 16472623 TI - Complementary stimulation of hepatobiliary transport and detoxification by rifampicin and ursodeoxycholic acid in humans. PMID- 16472624 TI - High-quality colonoscopies must be an integral part of screening and surveillance programs. PMID- 16472627 TI - Reducing the risk of interval colorectal neoplasia: high magnification chromoscopic colonoscopy may offer a key resource? PMID- 16472630 TI - Spectral processing by the peripheral auditory system: facts and models. PMID- 16472631 TI - Basic psychophysics of human spectral processing. PMID- 16472632 TI - Across-channel spectral processing. PMID- 16472633 TI - Speech and music have different requirements for spectral resolution. PMID- 16472634 TI - Non-linearities and the representation of auditory spectra. PMID- 16472635 TI - Spectral processing in the inferior colliculus. PMID- 16472636 TI - Neural mechanisms for spectral analysis in the auditory midbrain, thalamus, and cortex. PMID- 16472637 TI - Spectral processing in the auditory cortex. PMID- 16472638 TI - Processing of dynamic spectral properties of sounds. PMID- 16472639 TI - Representations of spectral coding in the human brain. PMID- 16472641 TI - Spectral information in sound localization. PMID- 16472640 TI - Spectral processing and sound source determination. PMID- 16472642 TI - Plasticity of spectral processing. PMID- 16472643 TI - Spectral processing in cochlear implants. PMID- 16472645 TI - Purification and biochemical properties of Rac1, 2, 3 and the splice variant Rac1b. AB - Rac proteins (Rac1, 1b, 2, 3) belong to the GTP-binding proteins (or GTPases) of the Ras superfamily and thus act as molecular switches cycling between an active GTP-bound and an inactive GDP-bound form through nucleotide exchange and hydrolysis. Like most other GTPases, these proteins adopt different conformations depending on the bound nucleotide, the main differences lying in the conformation of two short and flexible loop structures designated as the switch I and switch II region. The three distinct mammalian Rac isoforms, Rac1, 2 and 3, share a very high sequence identity (up to 90%), with Rac1b being an alternative splice variant of Rac1 with a 19 amino acid insertion in vicinity to the switch II region. We have demonstrated that Rac1 and Rac3 are very closely related with respect to their biochemical properties, such as effector interaction, nucleotide binding, and hydrolysis. In contrast, Rac2 displays a slower nucleotide association and is more efficiently activated by the Rac-GEF Tiam1. Modeling and normal mode analysis corroborate the hypothesis that the altered molecular dynamics of Rac2, in particular at the switch I region, may be responsible for different biochemical properties. On the other hand, our structural and biochemical analysis of Rac1b has shown that, compared with Rac1, Rac1b has an accelerated GEF-independent GDP/GTP-exchange and an impaired GTP-hydrolysis, accounting for a self-activating GTPase. This chapter discusses the use of fluorescence spectroscopic methods, allowing real-time monitoring of the interaction of nucleotides, regulators, and effectors with the Rac proteins at submicromolar concentrations and quantification of the kinetic and equilibrium constants. PMID- 16472646 TI - Biochemical analyses of the Wrch atypical Rho family GTPases. AB - The Rho family of GTPases comprises a major branch of the Ras superfamily of small GTPases. To date, at least 22 human members have been identified. However, most of our knowledge of Rho GTPase function comes from the study of the three classical Rho GTPases, RhoA, Rac1, and Cdc42. These Rho GTPases function as GDP/GTP-related binary switches that are activated by diverse extracellular signal-mediated stimuli. The activated GTPases then interact with downstream effectors to regulate cytoplasmic signaling networks that in turn regulate actin organization, cell cycle progression, and gene expression. Recently, studies have begun to explore the regulation and function of some of the lesser-known members of the Rho GTPase family. Wrch-1 (Wnt-regulated Cdc42 homolog-1) and the closely related Chp (Cdc42 homologous protein)/Wrch-2 protein comprise a distinct branch of the mammalian Rho GTPase family. Although both share significant sequence and functional similarities with Cdc42, Wrch proteins possess additional N- and C terminal sequences that distinguish them from the classical Rho GTPases (Cdc42, RhoA, and Rac1). We have determined that Wrch-1 and Wrch2 exhibit unusual GDP/GTP binding properties and undergo posttranslational lipid modifications distinct from those of the classical Rho GTPases. In this chapter, we summarize our experimental approaches used to characterize the biochemical properties of these atypical Rho GTPases. PMID- 16472647 TI - Purification of P-Rex1 from neutrophils and nucleotide exchange assay. AB - The P-Rex family of guanine-nucleotide exchange factors (GEFs) are activators of the small GTPase Rac (Donald et al., 2004; Rosenfeldt et al., 2004; Welch et al., 2002). They are directly regulated in vitro and in vivo by the lipid second messenger phosphatidylinositol (3,4,5)-triphosphate (PtdIns(3,4,5)P3) and by the betagamma subunits of heterotrimeric G proteins (Donald et al., 2004; Rosenfeldt et al., 2004; Welch et al., 2002). Activation by PtdIns(3,4,5)P3 occurs by means of the PH domain of P-Rex1 and activation by Gbetagamma subunits by means of the catalytic DH domain (Hill et al., 2005). P-Rex1 and P-Rex2 also contain two DEP and two PDZ protein interaction domains, as well as homology over their COOH terminal half to inositol polyphosphate 4-phosphatase (Donald et al., 2004; Welch et al., 2002). These domains, although not necessary for P-Rex1 activity in vitro, influence its basal and/or stimulated Rac-GEF activity, suggesting that their interaction with the DH/PH domain tandem is important for P-Rex1 function (Hill et al., 2005). P-Rex2B, a splice variant of P-Rex2, lacks the C-terminal half (Rosenfeldt et al., 2004). P-Rex1 was originally identified during a search for PtdIns(3,4,5)P3-dependent activators of Rac in neutrophils and purified to homogeneity from pig leukocyte cytosol, in which it is the major such activity (Welch et al., 2002). P-Rex1 is mainly expressed in neutrophils and regulates reactive oxygen species formation in these cells (Welch et al., 2002), whereas P Rex2 is expressed in a wide variety of tissues but not in neutrophils (Donald et al., 2004), and P-Rex2B is expressed in the heart (Rosenfeldt et al., 2004). This Chapter describes our methods for (1) the purification of endogenous P-Rex1 from pig leukocyte cytosol, (2) the production and purification of recombinant P-Rex proteins and their substrate GTPase Rac from Sf9 cells, and (3) the in vitro assay for measuring the GEF activities of native or recombinant P-Rex proteins. PMID- 16472648 TI - In vitro guanine nucleotide exchange activity of DHR-2/DOCKER/CZH2 domains. AB - Rho family GTPases regulate a large variety of biological processes, including the reorganization of the actin cytoskeleton. Like other members of the Ras superfamily of small GTP-binding proteins, Rho GTPases cycle between a GDP-bound (inactive) and a GTP-bound (active) state, and, when active, the GTPases relay extracellular signals to a large number of downstream effectors. Guanine nucleotide exchange factors (GEFs) promote the exchange of GDP for GTP on Rho GTPases, thereby activating them. Most Rho-GEFs mediate their effects through their signature domain known as the Dbl Homology-Pleckstrin Homology (DH-PH) module. Recently, we and others identified a family of evolutionarily conserved, DOCK180-related proteins that also display GEF activity toward Rho GTPases. The DOCK180-family of proteins lacks the canonical DH-PH module. Instead, they rely on a novel domain, termed DHR-2, DOCKER, or CZH2, to exchange GDP for GTP on Rho targets. In this chapter, the experimental approach that we used to uncover the exchange activity of the DHR-2 domain of DOCK180-related proteins will be described. PMID- 16472649 TI - Biochemical characterization of the Cool (Cloned-out-of-Library)/Pix (Pak interactive exchange factor) proteins. AB - The Cool (Cloned out of Library)/Pix (Pak interactive exchange factor) proteins have been implicated in a diversity of biological activities, ranging from pathways initiated by growth factors and chemoattractants to X-linked mental retardation. Initially discovered through yeast two-hybrid and biochemical analyses as binding partners for the Cdc42/Rac-target/effector, Pak (p21 activated kinase), the sequences for the Cool/Pix proteins revealed a DH (Dbl homology) domain. Because the DH domain is the limit functional unit for stimulating guanine nucleotide exchange on Rho family GTP-binding proteins, it was assumed that the Cool/Pix proteins would act as guanine nucleotide exchange factors (GEFs) for the Rho proteins. Of the three known isoforms, (p50Cool-1, p85Cool-1/beta-Pix, and 90Cool-2/alpha-Pix), only Cool-2/alpha-Pix has exhibited significant GEF activity. A number of experimental techniques have been used to characterize Cool-2, and in vitro analysis has revealed that its GEF activity is under tight control through intramolecular interactions involving several binding partners. Here we describe the biochemical methods used to study the Cool/Pix proteins and, in particular, the regulation of the GEF activity of Cool-2/alpha Pix. PMID- 16472650 TI - GEF and glucosylation assays on liposome-bound Rac. AB - Rac binds tightly to lipid membranes through a lipid modification. The influence of the lipid membrane environment on the multiple interactions of Rac has not been well documented. In this chapter, we detail a method to prepare geranyl geranylated Rac bound to liposomes of defined composition. With this method, one can dissect some lipid-protein interactions that facilitate the interaction of Rac with other proteins such as guanine nucleotide exchange factors and bacterial toxins. PMID- 16472651 TI - Phosphorylation of RhoGDI by p21-activated kinase 1. AB - Rho GTPase activation is partially regulated at the level of guanine nucleotide dissociation inhibitors, or GDIs. The binding of Rho GTPases to GDIs has been shown to dramatically reduce the action of guanine nucleotide exchange factors (GEFs) to initiate Rho GTPase activation. The GDI-GTPase complex thus serves as a major point of regulation of Rho GTPase activity and function. It is likely that specific mechanisms exist to dissociate individual members of the Rho GTPase family from cytosolic Rho GDI complexes to facilitate the activation process. Such dissociation would likely be tightly coupled to GEF-mediated guanine nucleotide exchange and membrane association of the activated GTPase, resulting in effector binding and functional responses. Accumulating evidence suggests that the phosphorylation of either the Rho GTPases themselves and/or phosphorylation of GDIs might serve as a mechanism for regulating the formation and/or dissociation of Rho GTPase-GDI complexes. Indeed, the selective release of Rac1 from RhoGDI complexes induced by the p21-activated kinase-regulated phosphorylation of RhoGDI has been reported. We describe here methods for the analysis of RhoGDI phosphorylation and regulation by p21-activated kinase 1 (Pak1). PMID- 16472652 TI - Purification of ARAP3 and characterization of GAP activities. AB - ARAP3 is a dual Arf and Rho GTPase activating protein (GAP) that was identified from pig leukocyte cytosol using a phosphatidylinositol-(3,4,5)-trisphosphate (PtdIns[3,4,5]P3) affinity matrix in a targeted proteomics study. ARAP3's domain structure includes five PH domains, an Arf GAP domain, three ankyrin repeats, a Rho GAP domain, and a Ras association domain. ARAP3 is a PtdIns(3,4,5)P3 dependent GAP for Arf6 both in vitro and in vivo. It acts as a Rap-GTP-activated RhoA GAP in vitro, and this activation depends on a direct interaction between ARAP3 and Rap-GTP; in vivo PtdIns(3,4,5)P3 seems to be required to allow ARAP3's activation as a RhoA GAP by Rap-GTP. Overexpression of ARAP3 in pig aortic endothelial (PAE) cells causes the PI3K-dependent loss of adhesion to the substratum and interferes with lamellipodium formation. This overexpression phenotype depends on ARAP3's intact abilities to bind PtdIns(3,4,5)P3, to interact with Rap-GTP, and to be a catalytically active RhoA and Arf6 GAP. PMID- 16472653 TI - Regulation of RhoGAP specificity by phospholipids and prenylation. AB - Among the key protein regulators of the various and numerous small GTPases are the GTPase activating proteins (GAPs). Experimental studies of some of the approximately 170 GAPs predicted by the human genome indicate that their catalytic GAP activity is regulated by a variety of mechanisms, including phosphorylation, protein-protein interactions, proteolysis, and interactions with lipids. Most reported biochemical studies to address the specificity of GAPs for particular GTPases have been conducted in vitro with bacterially produced GTPases. Thus, the potential influence of these various regulatory mechanisms in the context of GAP-GTPase specificity may be overlooked in such assays. Here, we present experimental studies that highlight the role of lipids in modulating the GTPase specificity for some of the Rho GAPs. We find that particular phospholipids can substantially alter the substrate "preference" for the p190 GAPs. We find that C-terminal prenylation of GTPases can influence the specificity of GAP interactions as well. These observations emphasize the limitations of standard in vitro GAP assays in definitively establishing the physiologically relevant GTPase targets for particular GAPs. PMID- 16472654 TI - Purification and activity of the Rho ADP-ribosylating binary C2/C3 toxin. AB - C3 exoenzyme from Clostridium limosum, specifically ADP-ribosylates and inactivates Rho GTPases, but not or much less than Rac and Cdc42. To bypass the poor cell accessibility of the exoenzyme, a chimeric fusion toxin was constructed consisting of C3 exoenzyme and the N-terminal adaptor domain of the enzyme component C2I of the actin-ADP-ribosylating Clostridium botulinum C2 toxin. This fusion toxin C2IN-C3 is transported into cells by interaction with the binding and translocation component (C2II) of C2 toxin. Purification and activity of the chimeric toxin is reported. PMID- 16472655 TI - Purification of TAT-C3 exoenzyme. AB - The Clostridium botulinum C3 exoenzyme has been an invaluable tool for the study of the biological functions of Rho GTPases. The C3 enzyme selectively catalyzes the ADP-ribosylation, and consequent inactivation, of RhoA, RhoB, and RhoC of the Rho GTPase protein family. Through the experimental use of C3, it has been possible to determine the contributions made by these signaling proteins to processes including the regulation of cell morphology, cell cycle progression, and gene transcription. Unlike bacterial toxins that have some means to attach to and/or enter cells, C3 does not have an element that facilitates efficient entry. As a result, numerous methods have been used to effectively deliver C3 into cells. One approach has been to engineer a recombinant C3 with an HIV TAT leader sequence that permits transduction of the protein across the plasma membrane. In this chapter, the purification and characterization of the recombinant TAT-C3 protein is described. PMID- 16472656 TI - Imaging and photobleach correction of Mero-CBD, sensor of endogenous Cdc42 activation. AB - This chapter details quantitative imaging of the Mero-CBD biosensor, which reports activation of endogenous Cdc42 in living cells. The procedures described are appropriate for imaging any biosensor that uses two different fluorophores on a single molecule, including FRET biosensors. Of particular interest is an algorithm to correct for fluorophore photobleaching, useful when quantitating activity changes over time. Specific topics include procedures and caveats in production of the Mero-CBD sensor, image acquisition, motion artifacts, shading correction, background subtraction, registration, and ratio imaging. PMID- 16472657 TI - Cdc42 and PI(4,5)P2-induced actin assembly in Xenopus egg extracts. AB - Xenopus egg cytoplasmic extracts have been used to study a variety of complex cellular processes. Given their amenability to biochemical manipulation and physiological balance of regulatory proteins, these extracts are an ideal system to dissect signal transduction pathways leading to actin assembly. We have developed methods to study Cdc42 and PI(4,5)P2-induced actin assembly in Xenopus egg extracts. In this chapter, we describe detailed procedures to prepare Xenopus egg extracts, Cdc42, and PI(4,5)P2 for use in actin assembly experiments. We also describe a fluorometric pyrene actin assay for quantitative kinetic analysis of actin polymerization and a microscopic rhodamine actin assay for quick measurement of actin rearrangements in extracts. Finally we provide a protocol for immunodepletion of proteins and discuss the use of immunodepletion and rescue experiments for functional analysis of components in the extracts. PMID- 16472658 TI - In vitro reconstitution of cdc42-mediated actin assembly using purified components. AB - In the accompanying chapter, we describe an in vitro system that uses Xenopus egg extracts to study actin assembly induced by phosphatidylinositol (4,5)bisphosphate (PIP2) and Cdc42. Biochemical fractionation and candidate screening experiments conducted in the extract system have identified the Arp2/3 complex, the N-WASP-WIP (or N-WASP-CR16) complex, and the Cdc42-binding protein Toca-1 as important mediators of PIP2- and Cdc42-actin signaling. Toward our ultimate goal of reconstituting an in vitro system that recapitulates the signaling properties observed in vivo, we then developed a purified actin assembly assay system consisting of the regulatory components that we discovered from extracts. In these assays, the stereotypical sigmoidal kinetics of actin polymerization are monitored by pyrene-actin fluorescence in the presence of defined recombinant or purified proteins, enabling the detailed study of mechanism and protein function. In this chapter, we describe the preparation of the components used in these purified actin assembly reactions, as well as the assay conditions under which we monitor actin polymerization kinetics in vitro. PMID- 16472659 TI - Biochemical analysis of mammalian formin effects on actin dynamics. AB - Formins are members of a conserved family of proteins, present in all eukaryotes, that regulate actin dynamics. Mammals have 15 distinct formin genes. From studies to date, surprising variability between these isoforms has been uncovered. All formins examined have several common effects on actin dynamics in that they: (1) accelerate nucleation rate; (2) alter filament barbed end elongation/depolymerization rates; and (3) antagonize capping protein. However, the potency of each effect can vary greatly between formins. In addition, a subset of formins binds tightly to filament sides and bundle filaments. Even isoforms that are closely related phylogenetically can display marked differences in their effects on actin. This chapter discusses several methods for examining formin function in vitro. We also discuss pitfalls associated with these assays. As one example, the effect of profilin on formin function is difficult to interpret by "pyrene-actin" polymerization assays commonly used in the field and requires assays that can distinguish between filament nucleation and filament elongation. The regulatory mechanisms for formins are not clear and certainly vary between isoforms. A subset of formins is regulated by Rho GTPases, and the assays described in this chapter have been used for characterization of this regulation. PMID- 16472660 TI - Formin proteins: purification and measurement of effects on actin assembly. AB - We describe methods for expressing and isolating formin proteins from a wide range of species and comparing quantitatively their effects on actin assembly. We first developed these procedures for purification of S. cerevisiae formins Bni1 and Bnr1 but have extended them to mammalian formins, including mouse mDia1 and mDia2 and human Daam1. Thus, the approach we describe should be universally applicable to the purification and analysis of formins from any eukaryote. Formins expressed in yeast rather than bacteria usually have improved solubility, yield, and actin assembly activity. Yields are 200-500 microg purified formin per liter of yeast culture. For some applications bacterial expression and purification is preferable, and these methods are also described. For expression of most formins, in either yeast or bacteria, we recommend using an amino terminal 6xHis affinity tag. Active FH1-FH2 containing fragments of the formins Bni1, Bnr1, mDia1, mDia2, and Daam1 are all digomeric. However, they nucleate actin filaments with variable efficiencies, as high as one actin filament per formin complex. In the last section, we outline fluorometric methods for measuring and quantitatively analyzing the in vitro activities of formins on actin nucleation and processive capping of actin filaments. PMID- 16472661 TI - Purification and kinase assay of PKN. AB - PKN is a serine/threonine protein kinase, which has a catalytic domain highly homologous to that of protein kinase C (PKC) in the carboxyl-terminal region and three repeats of the antiparallel coiled coil (ACC) domain in the amino-terminal region. Mammalian PKN has three isoforms each derived from different genes, PKN1 (PKNalpha/PRK1/PAK1), PKN2 (PRK2/PAK2/PKNgamma), and PKN3 (PKNbeta). PKN isoforms show different enzymatic properties and tissue distributions and have been implicated in various distinct cellular processes (reviewed in Mukai [2003]). This chapter discusses methods to prepare purified enzymes and to assay substrate phosphorylation activities. PMID- 16472662 TI - Purification and enzyme activity of ACK1. AB - The activated Cdc42 associated kinases (ACKs) are nonreceptor tyrosine kinases that are specific targets of Cdc42. To study the biochemical properties of ACK1, we expressed and purified the enzyme using the baculovirus/Sf9 cell system. This ACK1 construct contains (from N- to C-terminus) the kinase catalytic domain, SH3 domain, and Cdc42-binding CRIB domain. We describe enzyme activity assays based on synthetic peptide substrates. The best such substrate is a peptide derived from the site of ACK1-catalyzed phosphorylation of the Wiskott-Aldrich syndrome protein (WASP). Although the SH3 and CRIB domains of purified ACK1 are able to bind ligands (a polyproline peptide and Cdc42, respectively), the ligands did not stimulate in vitro tyrosine kinase activity. Purified ACK1 undergoes autophosphorylation at Tyr284, and autophosphorylation increases kinase activity. PMID- 16472663 TI - Direct activation of purified phospholipase C epsilon by RhoA studied in reconstituted phospholipid vesicles. AB - Phospholipase C-epsilon (PLC-epsilon) was shown recently to be a downstream effector of Rho GTPases, and we have used an in vitro phospholipid vesicle reconstitution system with purified proteins to show this regulation to be direct. This chapter describes high-level expression of a hexahistidine-tagged fragment of PLC-epsilon encompassing the catalytic core of the enzyme through the tandem RA domains by use of a recombinant baculovirus and High Five insect cells. The recombinant protein is purified to homogeneity using metal chelate affinity and size exclusion chromatography. The small GTPase RhoA also is expressed to high levels in a lipidated form after baculovirus expression in High Five cells and is purified to near homogeneity after detergent extraction and metal chelate affinity chromatography. The capacity of GTPgammaS-bound RhoA to stimulate the phospholipase activity of PLC-epsilon is assessed by reconstitution of the RhoA in mixed-detergent phospholipid micelles containing PtdIns(4,5)P2 substrate. PMID- 16472664 TI - Regulation of PLCbeta isoforms by Rac. AB - Small GTPases function as molecular switches, which transduce cellular signals from upstream regulators to downstream effectors in a guanine nucleotide dependent manner. Direct binding partners of small GTPases fall into four classes of both regulators and effectors that can be differentiated on the basis of the state of nucleotide required for binding. Here we describe a procedure for the rapid screening and quantitative assessment of direct interactions of the Rho family of small GTPases with effector molecules of the phospholipase Cbeta class of enzymes using surface plasmon resonance technology. The experimental format described is also readily adaptable toward characterizing guanine nucleotide dependent binding events of both small and heterotrimeric G proteins with various classes of GTPase regulatory proteins. PMID- 16472665 TI - Biochemical properties and inhibitors of (N-)WASP. AB - The Wiskott-Aldrich syndrome protein (WASP) is an effector of the Rho GTPase Cdc42 and a key component of signaling pathways that regulate the actin cytoskeleton. WASP is regulated by a number of ligands, and the mechanisms by which these act are beginning to be understood through detailed biochemical analyses. Here we describe the protocols we use to study WASP proteins, including the methods we use to purify signaling components and the assays we use to quantitatively characterize the biochemical and biophysical properties of WASP, its activation by Cdc42, and its inhibition by the small molecule wiskostatin. These methods have broad use within the WASP-related cytoskeletal-signaling pathway but are also applicable to investigations of other intramolecular and intermolecular interactions. PMID- 16472666 TI - The use of GFP to localize Rho GTPases in living cells. AB - The green fluorescent protein (GFP) of the jellyfish Aequorea victoria has revolutionized the study of protein localization and dynamics. GFP fusions permit analysis of proteins in living cells and offer distinct advantages over conventional immunofluorescence. Among these are lower background, higher resolution, robust dual color colocalization, and avoidance of fixation artifacts. In the case of Ras and Rho family proteins, GFP fusions have allowed breakthroughs in the understanding of how CAAX proteins are targeted to specific cell membranes and how signaling at different membranes can result in different cellular responses. GFP-tagged Rho proteins have also been informative in analyzing the interactions with the cytosolic chaperone, RhoGDI. The major disadvantages of studying GFP fusion proteins is that they are generally overexpressed relative to endogenous proteins, and the GFP tag can, in principle, affect protein function. Fortunately, in the case of Ras and Rho family proteins, a GFP tag at the N terminus seems to have little effect on protein targeting and function. Nevertheless, it is prudent to confirm GFP fusion protein data with the study of the endogenous protein. This chapter describes the tagging of Rho proteins with GFP and the analysis of GFP-Rho protein localization by epifluorescence and confocal microscopy. It further describes methods of analyzing endogenous Rho proteins as confirmation of data acquired using GFP-Rho fusion proteins. These techniques will be useful for anyone studying Rho protein function and are widely applicable to many cell types and signal transduction systems. PMID- 16472667 TI - Analysis of the spatiotemporal activation of rho GTPases using Raichu probes. AB - GFP-based FRET probes that can visualize local activity changes in Rho GTPases in living cells are now available for examining the spatiotemporal regulation of these proteins. We previously developed FRET probes for Rho (and Ras) GTPases and collectively designated them "Ras and interacting protein chimeric unit" (Raichu) probes. In this chapter, we describe the principles and strategies used to develop Raichu-type FRET probes for Rho-family GTPases. The procedures for characterizing candidate probes, setting up the imaging system, and image acquisition/processing are also explained. An optimal FRET probe should: (1) have a wide dynamic range (i.e., a high sensitivity); (2) demonstrate high fluorescence intensity (i.e., a high signal-to-noise ratio); (3) show target specificity; and (4) cause minimal perturbation of endogenous signaling cascades. Although improvements of FRET probes should be executed in a trial-and-error manner, we provide practical tips for their optimization. In addition, some experimental results are presented to illustrate the expanding number of fields for the application of Raichu-RhoA/Rac1/Cdc42, and the advantages and disadvantages of Raichu probes are discussed. PMID- 16472668 TI - Measurement of activity of rho GTPases during mitosis. AB - The members of Rho-family GTPases regulate progression through mitosis. Rho induces the contractile ring at the equatorial cortex of the dividing cell, and thus works as a molecular switch between nuclear division and cytokinesis. Cdc42 regulates the progression from prometaphase to metaphase by stabilizing microtubule attachment to the kinetochore. These results suggest that Rho GTPases are activated at specific points in mitosis and regulate each step. Here we describe the methods to analyze the activity and regulation of Rho GTPases during mitosis. PMID- 16472669 TI - Inhibition of rho GTPases by RNA interference. AB - Selective down-modulation or silencing of individual members of the Rho-GTPase family is now practical using RNA interference. Transfection of mammalian cells with an individual siRNA duplex or siRNA pools can suppress expression of a specific isoform to understand its function. By adjusting the dose of siRNA, intermediate levels of suppression can be attained to test the biological role of different levels of a GTPase such as Rac. Nevertheless, there are significant potential pitfalls, including "off-target" effects of the siRNA on other genes. Besides demonstrating successful, noncytotoxic suppression of protein and activity levels of a specific GTPase, controls are essential to establish specificity. In this chapter, we provide methods for selective knockdown of expression by siRNA and confirmation of the effectiveness of Rho GTPase silencing, as well as descriptions and some examples of controls for specificity that include evaluations of dose-response, negative and positive controls, GTPase specificity, confirmation by using more than one siRNA for the same gene, rescue by a mutated siRNA-resistant cDNA encoding the target gene, and complementary supporting evidence. Selective silencing of specific Rho family GTPases should provide increasing insight into the regulatory and functional roles of each isoform in a wide variety of biological processes. PMID- 16472670 TI - RNA interference techniques to study epithelial cell adhesion and polarity. AB - Polarized epithelial cells are characterized by distinct plasma membrane domains and asymmetrical distribution of cell surface proteins and lipids. In vertebrates, tight junctions act as a fence between the apical and basolateral domains. Although many of the key components of the polarity machinery have been identified, their functions in cell polarization and junction formation remain to be determined. With the rapid improvement of the RNA interference (RNAi) technique, it is now possible to silence the expression of these polarity proteins in mammalian cells and to systematically analyze their distinct roles in orchestrating the polarization program. Here we describe approaches to achieve specific gene suppression in MDCK cells, a well-established cell culture model of canine kidney cells. We discuss the potential challenges and problems associated with the RNAi technique and describe basic protocols for suppressing gene expression using a vector-based short hairpin RNA (shRNA) expression system coupled with nucleofection. PMID- 16472671 TI - Nucleofection of primary neurons. AB - Efficient gene transfer is an important tool for the study of neuronal function and biology. This has proved difficult and inefficient with traditional transfection strategies, which can also be fairly toxic, whereas viral-mediated gene transfer, although highly efficient, is often time-consuming. The recently developed Amaxa Nucleofector technology, based on electroporation with preset parameters in a cell-type-specific solution, enables direct delivery of DNA, small interfering (si)RNA oligonucleotides and siRNA vectors into the cell nucleus. This strategy results in reproducible, rapid, and efficient transfection of a broad range of cells, including primary neurons. Nucleofected neurons survive for up to 3 weeks and remain functional. We are currently using this transfection method to examine the contribution of Rho GTPase signaling pathways in the establishment of neuronal polarity, neuronal migration, and neurite outgrowth. Here, we describe three protocols to efficiently nucleofect rat cerebellar granule, cortical, and hippocampal neurons. PMID- 16472672 TI - Dock180-ELMO cooperation in Rac activation. AB - Dock180 superfamily of proteins has been recently identified as novel, unconventional guanine nucleotide exchange factors (GEF) for Rho-family GTPases. Unlike most other GEFs for Rho-family GTPases, Dock180 family members do not contain the characteristic Dbl homology (DH) domain. Instead, they use a conserved "Docker" or "CZH2" domain to mediate the nucleotide exchange on Rho family GTPases. The Dock180 family members are evolutionarily conserved from worms to mammals. They play critical roles in a number of biological processes essential for the normal development of entire organisms, as well as for the physiological responses of these organisms, including removal of apoptotic cells and directed cell migration in C. elegans; myoblast fusion, and dorsal closure in Drosophila; lymphocyte migration, T-cell activation, tumor metastasis, HIV infection, and development of neuronal degenerative diseases in mammals. All these biological activities of the Dock180 family members have been linked to their ability to activate their specific GTPase substrate. At least four members of the Dock180 family bind to another evolutionarily conserved protein ELMO to optimally activate the Rac GTPase. The best characterized is the Rac activation by the Dock180-ELMO complex. ELMO modulates the Rac activation by Dock180 by means of at least three distinct mechanisms: helping Dock180 stabilize Rac in its nucleotide-free transition state; relieving a self-inhibition of Dock180; and targeting Dock180 to the plasma membrane to gain access to Rac. Thus, Dock180 and ELMO function together as a bipartite GEF to optimally activate Rac on upstream stimulation to mediate the engulfment of apoptotic cells and cell migration. PMID- 16472673 TI - Rho GTPase activation by cell-cell adhesion. AB - Cell-cell adhesion can occur in a calcium-dependent or calcium-independent manner, depending on the type of receptor involved. Establishment of cell contacts by either type of cell-cell adhesion (calcium-dependent or calcium independent) has been shown to activate Rho GTPases in different cells. In this chapter, we describe the method used to assess the activation of Rho GTPases by cadherins, the prototype calcium-dependent adhesion receptor in epithelial cells. We cover the optimal cell culture conditions and controls to ensure that the activation of the GTPases is specifically triggered by the formation of cadherin dependent cell-cell contacts. Controls described herein determine the specificity of activation of Rho proteins with respect to cadherin adhesion and exclude the contribution of other adhesive receptors, calcium-signaling, cell spreading, and migration. Although we focus on cadherin receptors and normal human keratinocytes as our model system, the methods described can be easily adapted to other adhesion receptors and different cell types. PMID- 16472674 TI - Activation of Rap1, Cdc42, and rac by nectin adhesion system. AB - Nectin is an immunoglobulin-like cell-cell adhesion molecule and forms adherens junctions cooperatively with cadherin. Trans-interaction of nectin induces activation of Rap1, Cdc42, and Rac small G proteins. The activity of these small G proteins can be analyzed by the pull-down assay using GST-Ral-GDS fusion protein for Rap1 and GST-PAK-CRIB for Cdc42 and Rac. The fluorescent resonance energy transfer (FRET) system is also available to spatially and temporally detect the activity of these small G proteins in the living cells. In addition to these assays, the activity of Cdc42 and Rac is indirectly, but easily, evaluated by the cell-spreading assay to examine formation of filopodia and lamellipodia, respectively. To clearly explore the effect of trans-interacting nectin on the small G proteins, we use L fibroblasts stably expressing nectin-1 (nectin-1-L cells) and the extracellular domain of nectin-3 fused to human IgG Fc (Nef-3). Treatment of nectin-1-L cells with Nef-3 remarkably increases both the amount of the GTP-bound form and the FRET efficiency of all Rap1, Cdc42, and Rac small G proteins. In the cell-spreading assay, Cdc42 and Rac activated in this way promote the formation of filopodia and lamellipodia, respectively. Here, we focus on how the activity of Cdc42 and Rac induced by trans-interacting nectin is examined by use of the pull-down and the cell-spreading assays. PMID- 16472675 TI - Analysis of activated GAPs and GEFs in cell lysates. AB - An assay was developed that allows the precipitation of the active pools of Rho GEFs, Rho-GAPs, or effectors from cell or tissue lysates. This assay can be used to identify GEFs, GAPs, and effectors involved in specific cellular pathways to determine their GTPase specificity and to monitor the temporal activation of GEFs and GAPs in response to upstream signals. PMID- 16472676 TI - Degradation of RhoA by Smurf1 ubiquitin ligase. AB - The Rho family of small GTPases plays a key role in the dynamic regulation of the actin cytoskeleton that underlies various important cellular functions such as shape changes, migration, and polarity. We found that Smurf1, a HECT domain E3 ubiquitin ligase, could specifically target RhoA but not Cdc42 or Rac1 for degradation. Smurf1 interacts with the dominant inactive form of RhoA, RhoA N19, which binds constitutively to guanine nucleotide exchange factors (GEFs) in vivo. Smurf1 also interacts directly with either nucleotide-free or GDP-bound RhoA in vitro; however, loading with GTPgammaS inhibits the interaction. RhoA is ubiquitinated by wild-type Smurf1 but not the catalytic mutant of Smurf1 (C699A) in vivo and in vitro, indicating that RhoA is a direct substrate of Smurf1. In this chapter, we summarize the systems and methods used in the analyses of Smurf1 regulated RhoA ubiquitination and degradation. PMID- 16472677 TI - Ubiquitin-mediated proteasomal degradation of Rho proteins by the CNF1 toxin. AB - The CNF1 toxin is produced by uropathogenic and meningitis-causing Escherichia coli. CNF1 catalyzes the constitutive activation of Rho proteins by deamidation. The threshold of activation of Rho proteins by CNF1 is, however, attenuated because of a concomitant decrease of their cellular levels. Depletion of activated-Rac1 is catalyzed by ubiquitin-mediated proteasomal degradation. Consequently, we show by effector-binding pull-down that co-treatment of intoxicated cells with the MG132 proteasome-inhibitor results in a higher level of activation of Rac, as well as RhoA and Cdc42. We show that CNF1 induces the transient recruitment of Rho proteins to cellular membranes. Interestingly, at the difference of Rac and Cdc42, the inhibition of the proteasome during CNF1 treatment does not result in a significant accumulation of RhoA to cellular membranes. Using an in vivo ubiquitylation assay, we evidence that mutation of the geranylgeranyl acceptor cysteine of Rac1 (Rac1C189G) abolished the sensitivity of permanently activated-Rac1 to ubiquitylation, whereas Rac1C189G remained able to bind to the effector-binding domain of p21-PAK. Collectively, these results indicate that association with the cellular membranes is a necessary step for activated-Rac1 ubiquitylation. PMID- 16472678 TI - Regulation of superoxide-producing NADPH oxidases in nonphagocytic cells. AB - The membrane-integrated protein gp91phox functions as the catalytic center of the superoxide-producing phagocyte NADPH oxidase. Recent studies have identified homologs of gp91phox in nonphagocytic cells, which constitute the NADPH oxidase (Nox) family. Activation of the Nox oxidases leads to production of reactive oxygen species (ROS), thereby participating in a variety of biological events, such as host defense, hormone biosynthesis, and signal transduction. The activity of the Nox enzymes is regulated by various proteins, including the small GTPase Rac; regulatory mechanisms differ dependent on the type of the Nox proteins. For example, an oxidase activator (p47phox or Noxo1) and an oxidase activator (p67phox or Noxa1) are absolutely required for superoxide production by gp91phox and Nox1, but not by Nox3. Rac, albeit probably dispensable to the Nox3 activity, plays an essential role in activation of gp91phox. Thus, functional reconstitution of Nox systems is crucial for the study of Nox regulation. Here we describe a basic method for the reconstitution of Nox systems by expression of oxidase proteins in transfectable cells. PMID- 16472679 TI - Activation of MEKK1 by Rho GTPases. AB - Mammalian MAP/ERK kinase kinase 1 (MEKK1) is MAP kinase kinase kinase (MAP3K) that is a crucial regulator of many cellular signaling cascades. One of the most important physiological functions of MEKK1 is its ability to regulate cell migration, because MEKK1 null mice are defective in eyelid closure. MEKK1 exhibits its signaling activity through interaction with a large array of cellular factors, including several proteins that are known to play central roles in controlling cell movement and motility. We have recently identified an interaction between MEKK1 and RhoA. This interaction occurs between the GTP bound, active form of RhoA and the amino terminal region of MEKK1 that harbors a PHD domain with E3 ubiquitin ligase activity. RhoA-GTP activates MEKK1 in vitro and in cells. Here we describe in detail the assay methods for RhoA activation of MEKK1, including preparation of recombinant proteins and proteins immunoprecipitated from cells, pretreatment of proteins, and assay conditions. We also briefly explain the methods and conditions we use to identify the interaction between MEKK1 and RhoA in yeast and in mammalian cells. PMID- 16472680 TI - Activation of the apoptotic JNK pathway through the Rac1-binding scaffold protein POSH. AB - The JNKs (c-Jun N-terminal protein kinases) play important roles in a variety of physiological and pathological functions including induction of apoptosis. A major pathway by which JNKs are activated in response to apoptotic stimuli includes the GTP*Rac1-binding scaffold protein POSH (plenty of SH3s). POSH acts as a scaffold for binding and autoactivation of the MLK family of MKKK proteins, which in turn phosphorylate and activate the MKK family members MKK4 and 7, which in turn phosphorylate JNKs. In this chapter, we describe methods and techniques that have been successfully used to study the POSH-dependent apoptotic JNK pathway. Use of these techniques may lead to a better understanding of the components of this pathway and of how it is suppressed in viable cells and rapidly activated in response to apoptotic stimuli. PMID- 16472682 TI - Activation of Rho and Rac by Wnt/frizzled signaling. AB - Wnt/Frizzled (Fz) signaling controls developmental, physiological, and pathological processes by means of several distinct pathways. Wnt/Fz activation of the small GTPases Rho and Rac has emerged as a key mechanism that regulates cell polarity and movements during vertebrate gastrulation. In this chapter, we describe biochemical assays for the detection of Wnt/Fz activation of Rho and Rac in both mammalian cells and Xenopus embryo explants. PMID- 16472681 TI - Quantification of isozyme-specific activation of phospholipase C-beta2 by Rac GTPases and phospholipase C-epsilon by Rho GTPases in an intact cell assay system. AB - Phospholipase C (PLC) catalyzes the hydrolysis of PtdIns(4,5)P2, which results in both formation of the second messengers Ins(1,4,5)P3 and diacylglycerol and alteration in the membrane association and/or activity of PtdIns(4,5)P2-binding proteins. The existence of 13 different PLC isozymes suggests multiple mechanisms of regulation of inositol lipid signaling, and the recent realization that Rho family GTPases directly bind and activate certain PLC isozymes has added to this potential diversity of inositol lipid-related signal transduction. With the goal of delineating a less labor-intensive method for quantification of intracellular inositol phosphate production, we have applied a commercially available yttrium silicate RNA binding resin selective for inositol phosphates to develop a high throughput inositol phosphate scintillation proximity assay (SPA). We highlight the utility of this assay using COS-7 cells robotically transfected in a 96-well format. This method is readily applied to quantify activation of PLC by receptors and G proteins, and we illustrate here the selective activation of PLC-beta2 by Rac but not by Rho GTPases and the selective activation of PLC-epsilon by Rho but not Rac GTPases. PMID- 16472683 TI - Fluorescent assay of cell-permeable C3 transferase activity. AB - C3 transferase has been extensively used in the investigation of Rho signaling. It acts as a Rho antagonist by covalently attaching an ADP-ribose group to RhoA, B, and C proteins. Cell-permeable C3 fusion proteins have been developed so that lower doses of C3 can be used. We describe a simple and fast fluorescence-based assay to evaluate the enzymatic activity of cell-permeable C3 proteins purified from Escherichia coli. The assay measures glycohydrolase (GH) activity of C3 that cleaves NAD+ into ADP-ribose and nicotinamide. Results from the GH activity correlate with other tests carried out in tissue culture cells such as neurite outgrowth or ADP ribosylation of RhoA. This method provides reliable measurements of the activity of permeable C3 proteins or other C3-related proteins. PMID- 16472684 TI - Use of TIRF microscopy to visualize actin and microtubules in migrating cells. AB - Total internal reflection fluorescence (TIRF) is the technique of choice to visualize and quantify cellular events localized at the basal plasma membrane of adherent cells. By selectively illuminating the first 200 nm above the basal membrane, it allows maximal resolution in the vertical z-axis. In this chapter, I describe a prism-based TIRF setup and the procedures to visualize the actin and microtubule cytoskeleton in migrating astrocytes. TIRF microscopy provides quantitative information on the organization of the cytoskeleton in both fixed and live migrating cells. PMID- 16472685 TI - Inhibition of ROCK by RhoE. AB - RhoE belongs to the Rnd subfamily of small Rho-related GTP-binding proteins. Similar to other Rho proteins, RhoE regulates actin cytoskeleton dynamics. Expression of RhoE induces loss of actin stress fibers, and it also increases cell migration speed. In part, this is due to RhoE interaction with the RhoA effector ROCK I, a serine/threonine kinase that regulates the formation and contractility of stress fibers. Interestingly, RhoE does not interact with the highly homologous kinase ROCK II. RhoE binding inhibits ROCK I from phosphorylating its downstream target myosin light chain phosphatase, thus increasing the activity of the phosphatase to dephosphorylate myosin II, which results in reduced actomyosin contractility. RhoE itself is phosphorylated by ROCK I, and this may enhance RhoE regulation of ROCK I function. This chapter describes the method used for studying ROCK inhibition by RhoE. PMID- 16472686 TI - Conditional regulation of a ROCK-estrogen receptor fusion protein. AB - The Rho-associated kinases ROCK I and ROCK II are serine/threonine kinases that play central and critical roles in regulating the actin cytoskeleton. Activation of ROCK proteins contributes positively to the phosphorylation of myosin II light chains (MLC), myosin ATPase activity, stabilization of filamentous actin, and coupling of the actin-myosin filaments to the plasma membrane, thereby leading to the increased actin-myosin force generation and contractility. We have constructed a conditionally-activated form of ROCK II (called ROCK:ER) by fusing the ROCK II kinase domain to the estrogen receptor hormone-binding domain. In this chapter, we describe the construction and characterization of this regulatable ROCK:ER fusion protein. PMID- 16472687 TI - Rational design and applications of a Rac GTPase-specific small molecule inhibitor. AB - Rac GTPases are involved in the regulation of multiple cell functions and have been implicated in the pathology of certain human diseases. Dominant negative mutants of Rac have been the tool of choice in studying Rac function in cells. Given the difficulty of introducing high concentrations of the Rac mutants into primary cells and nonspecific effects of the mutants on Rho guanine nucleotide exchange factor (GEF) activities, it is desirable to develop small molecule inhibitors that could specifically inhibit Rac activities. Here we describe the rational design, characterization, and applications of a first-generation Rac specific small molecule inhibitor. On the basis of the structure-function information of Rac interaction with GEFs, in a computer-based virtual screening we have identified NSC23766, a highly soluble and membrane permeable compound, as a specific inhibitor of a subset of GEF binding to Rac and, therefore, Rac activation by these GEFs. In fibroblast cells, NSC23766 inhibited Rac1 GTP loading without affecting Cdc42 or RhoA activity and suppressed cell proliferation induced by a Rac GEF Tiam1. It has little effect on cell growth induced by a constitutively active Rac1 mutant. In addition, NSC23766 inhibited: (1) the anchorage-independent growth and invasion phenotypes of human prostate cancer PC-3 cells; (2) Rac activation and Rac-dependent aggregation of platelets stimulated by thrombin; and (3) Rac1 and Rac2 activities of hematopoietic stem/progenitor cells and induced their mobilization from mouse bone marrow to peripheral blood. Thus, NSC23766 is a lead small molecule inhibitor of Rac activity and could be useful for studying Rac-mediated cellular functions and for modulating pathological conditions in which Rac-deregulation may play a role. PMID- 16472688 TI - In vitro assay of primary astrocyte migration as a tool to study Rho GTPase function in cell polarization. AB - Rho GTPases are key players in cell migration. The contribution of Rho, Rac, and Cdc42 to the regulation of the actin and microtubule cytoskeletons is essential for membrane protrusion and cell retraction (Etienne-Manneville and Hall, 2002). The polarization of these protrusive and retracting activities in a migrating cell is also under the control of Rho GTPases, in particular Cdc42 (Nobes and Hall, 1999). In vitro study of cell migration has shown that Cdc42 activity is required for polarized cell migration in several cell types, including fibroblasts, neutrophils, macrophages, and astrocytes (Allen et al., 1998; Etienne-Manneville, 2004; Etienne-Manneville and Hall, 2001; Palazzo et al., 2001; Srinivasan et al., 2003). Using scratch-induced migration assay, we have previously used primary astrocytes as a tool to study the molecular mechanisms controlling cell polarization at the onset of migration (Etienne-Manneville and Hall, 2001, 2003). On scratching of the monolayer, astrocytes polarize perpendicularly to the scratch to migrate and close the wound. Astrocyte polarization is characterized by the formation of a protrusion in the direction of migration, the elongation of the microtubules that fill the protrusion, and the reorientation of the centrosome, which serves as a microtubule-organizing center toward the direction of migration. This in vitro migration assay allows us to simultaneously investigate the mechanisms controlling cell migration, cell protrusion, and cell polarization. Primary astrocytes, although more constraining, provide a more physiological model than immortalized cell lines. Moreover, astrocyte culture can be obtained in a large number and, therefore, also allows biochemical analysis. Here I describe the procedure by which we can obtain and purify primary rat astrocytes and the different assays we have previously used to analyze the role of Rho GTPases and their downstream targets in cell migration and polarization. PMID- 16472689 TI - Real-time centrosome reorientation during fibroblast migration. AB - The centrosome is positioned between the nucleus and the leading edge of many types of migrating cells. Cdc42 regulates this centrosome reorientation through its effectors Par6 and MRCK. Using time-lapse microscopy of live cells, the mechanisms and kinetics of centrosome reorientation can be studied. In this chapter, we describe a modification in the standard wound healing assay that allows the study of signaling pathways involved in centrosome reorientation and other polarization events that occur before cell migration. We also describe a method for visualization of centrosome reorientation by time-lapse microscopy using NIH 3T3 fibroblasts stably transfected with GFP-tubulin. PMID- 16472690 TI - Lentiviral delivery of RNAi in hippocampal neurons. AB - The breakthrough discovery that double-stranded RNA of 21 nucleotides in length (referred to as short or small interfering RNA; siRNA) can trigger sequence specific gene silencing in mammalian cells has led to the development of a powerful new approach to study gene function (Dillon et al., 2005; Dykxhoorn et al., 2003; Elbashir et al., 2001; Hannon et al., 2004). Effective delivery of siRNA molecules into target cells or tissues is critical for successful RNA interference (RNAi) application. Here, we describe the use of human immunodeficiency virus type 1 (HIV-1)-based lentiviral vectors for delivery of short hairpin RNA (shRNA), a precursor of siRNA, into primary neurons to suppress gene expression. Major advantages of lentiviral vectors are their ability to transduce nondividing cells and to confer long-term expression of transgenes. This chapter covers selection of short hairpin sequences, vector design, production of lentiviral supernatants, transduction of dissociated primary hippocampal neurons, and testing the effectiveness of shRNA-mediated silencing. PMID- 16472691 TI - Methods for studying neutrophil chemotaxis. AB - Chemotaxis is a fascinating biological process, through which a cell migrates along a shallow chemoattractant gradient that is less than 5% difference between the anterior and posterior of the cell. Chemotaxis is composed of two independent, but interrelated processes--motility and directionality--both of which are regulated by extracellular stimuli and chemoattractants; small GTPases have been shown to be involved. In this chapter, the methods that are used to prepare mouse neutrophils and study their chemotactic behaviors and morphological and biochemical changes in response to chemoattractant stimulation are described. PMID- 16472692 TI - Measurement of epidermal growth factor receptor turnover and effects of Cdc42. AB - Ligand-induced degradation represents an essential component of the overall regulation of EGF receptor (EGFR)-coupled signal transduction. Following activation, EGFRs are monoubiquitinated, subsequently sorted by ubiquitin interaction-based sorting machinery, and transported to multivesicular bodies (MVBs) and lysosomes for degradation. The Rho-family small G-protein, Cdc42, has been implicated in the regulation of EGFR degradation. Here we describe routine methods for assaying EGFR endocytosis and degradation. In addition, we have introduced procedures for determining the effects of Cdc42 and its downstream targets, in particular, ACK (Activated Cdc42-associated Kinase) and p85Cool-1 (Cloned out of library)/Pix (for Pak-interactive exchange factor), on EGFR degradation. PMID- 16472694 TI - An in vitro model to study the role of endothelial rho GTPases during leukocyte transendothelial migration. AB - Tissue injury induces release of cytokines that stimulate the expression of adhesion receptors in the endothelial wall of neighboring vessels. These endothelial receptors recruit leukocytes from the bloodstream and facilitate their transendothelial migration (TEM) toward the inflamed area. The molecules involved in leukocyte-endothelium interaction and TEM have been studied in vivo and in vitro over the past 20 years. Human umbilical vein endothelial cells (HUVECs) are a popular in vitro model to analyze TEM, and have been used to investigate the role of Rho GTPases in this process. Here we describe methods to activate HUVECs, to investigate Rho GTPase-activation by the endothelial adhesion receptor ICAM-1, and to inhibit or activate Rho GTPases using C3 transferase or adenoviruses coding for dominant negative or constitutive active Rho GTPases. Finally, we describe how to image and quantitate leukocyte TEM by digital time lapse microscopy. PMID- 16472693 TI - Tumor cell migration in three dimensions. AB - In almost all physiological and pathological situations, cells migrate through three-dimensional environments, yet most studies of cell motility have used two dimensional substrates. It is clear that two-dimensional substrates do not mimic the in vivo environment accurately, and recent work using three-dimensional environments has revealed many different mechanisms of cell migration (Abbott, 2003; Sahai and Marshall, 2003; Wolf et al., 2003). This chapter will describe methods for generating three-dimensional matrices suitable for studying cell motility, methods for imaging the morphology of motile cells in situ, and methods for quantifying cell migration through three-dimensional environments. PMID- 16472695 TI - Analysis of a mitotic role of Cdc42. AB - Rho GTPases including Rho, Rac, and Cdc42 determine the cell shape by regulating the actin and microtubule dynamics. Through these actions on cytoskeleton, Rho GTPases also regulate cell cycle progression. Specifically, Rho, Rac, and Cdc42 regulate G1-S progression, and Rho regulates cytokinesis. However, involvement of these GTPases in nuclear division has not been definitely shown. This seems to be due to the lack of standard procedures examining mitosis-specific functions of these GTPases. Recently, we have found a mitosis-specific role of Cdc42 by enrichment of cells in mitosis by cell cycle synchronization and interfering with functions of Rho GTPases. This chapter describes the procedures we used in these studies. PMID- 16472697 TI - Morphological and biochemical analysis of Rac1 in three-dimensional epithelial cell cultures. AB - Rho GTPases are critical regulators of epithelial morphogenesis. A powerful means to investigate their function is three-dimensional (3D) cell culture, which mimics the architecture of epithelia in vivo. However, the nature of 3D culture requires specialized techniques for morphological and biochemical analyses. Here, we describe protocols for 3D culture studies with Madin-Darby Canine Kidney (MDCK) epithelial cells: establishing cultures, immunostaining, and expressing, detecting, and assaying Rho proteins. These protocols enable the regulation of epithelial morphogenesis to be explored at a detailed molecular level. PMID- 16472696 TI - Plexin-induced collapse assay in COS cells. AB - Semaphorins are a family of growth cone guidance molecules. When associated with their receptors and coreceptors, plexins and neuropilins, they act as chemorepellents for an extensive range of neuronal populations. The prototypic semaphorin, Sema3A, has a potent inhibitory effect on sensory axons emanating from dorsal root ganglia. This has formed the basis of the most famous assay for semaphorin activity, the chick dorsal root ganglia collapse assay. Recently, a heterologous, highly tractable assay has been used to investigate semaphorin signaling. In this system, the binding of recombinant semaphorins to COS cells expressing plexins and neuropilins induces a morphological collapse that may correlate with growth cone collapse. This chapter describes the optimization of this assay and outlines the subtle differences required to enable Sema3A-Fc and Sema4D-Fc to induce identical collapse phenotypes in COS cells expressing Plexin A1 and neuropilin-1, or Plexin-B1, respectively. PMID- 16472698 TI - Using three-dimensional acinar structures for molecular and cell biological assays. AB - Epithelial cells grown in three-dimensional matrix allows one to investigate biological processes in a setting that is closer to in vivo conditions than those obtained by growing cells on plastic culture dishes. Here we outline procedures that allow one to investigate molecular mechanisms that regulate formation and disruption of three-dimensional epithelial structures. PMID- 16472699 TI - TC10 and insulin-stimulated glucose transport. AB - Insulin stimulates glucose uptake in insulin-responsive tissues by means of the translocation of the glucose transporter GLUT4 from intracellular sites to the plasma membrane. Two pathways are required, one involving activation of a phosphatidylinositol 3-kinase (PI 3-kinase) and downstream protein kinases, and one involving activation of the Rho-family GTPase TC10. TC10 activation by insulin is catalyzed by the exchange factor C3G, which is translocated to lipid rafts along with its binding partner CrkII as a consequence of Cbl tyrosine phosphorylation by the insulin receptor. This activation of TC10 is dependent on localization of TC10 in the lipid raft subdomains of the plasma membrane. We describe experimental approaches using the insulin-responsive cell line 3T3-L1 adipocytes to study the role of TC10 in insulin-stimulated glucose transport. PMID- 16472701 TI - In vitro assembly of filopodia-like bundles. AB - A breakthrough in understanding the mechanism of lamellipodial protrusion came from development of an in vitro model system, namely the rocketing movement of microbes and activated beads driven by actin comet tails (Cameron et al., 1999, 2000; Loisel et al., 1999; Theriot et al., 1994). As a model for investigation of the other major protrusive organelle, the filopodium, we developed in vitro systems for producing filopodia-like bundles (Vignjevic et al., 2003), one of which uses cytoplasmic extracts and another that reconstitutes like-like bundles from purified proteins. Beads coated with Arp2/3-activating proteins can induce two distinct types of actin organization in cytoplasmic extracts: (1) comet tails or clouds displaying a dendritic array of actin filaments and (2) stars with filament bundles radiating from the bead. Actin filaments in star bundles, like those in filopodia, are long, unbranched, aligned, uniformly polar, and grow at the barbed end. Like filopodia, star bundles are enriched in fascin and lack Arp2/3 complex and capping protein. Similar to cells, the transition from a dendritic (lamellipodial) to a bundled (filopodial) organization is induced by depletion of capping protein, and add-back of this protein restores the dendritic mode. By use of purified proteins, a small number of components are sufficient for the assembly of filopodia-like bundles: WASP-coated beads, actin, Arp2/3 complex, and fascin. On the basis of analysis of this system, we proposed a model for filopodial formation in which actin filaments of a preexisting dendritic network are elongated by inhibition of capping and subsequently cross-linked into bundles by fascin. PMID- 16472700 TI - GTPases and the control of neuronal polarity. AB - Neurons are probably the most highly polarized cell type and typically develop a single axon and several dendrites. The establishment of a polarized morphology and the functional specialization of axonal and dendritic compartments are essential steps in the differentiation of neurons. Primary cultures of dissociated hippocampal neurons are a widely used system to study the development of neuronal differentiation. In this article, we will describe gain-of-function and loss-of-function approaches that allow us to analyze the role of GTPases in neuronal differentiation. PMID- 16472703 TI - Second-line treatment options in advanced non-small cell lung cancer: current status. AB - Despite the emergence of several antineoplastic drugs with activity in advanced non-small cell lung cancer (NSCLC), real progress with first-line therapy has been limited in the last decade. It has proven difficult to demonstrate important differences between third generation two-drug combinations. In contrast, progress with second-line therapy has been significant, especially in the past few years, with pemetrexed showing equivalent activity to docetaxel but with substantially less toxicity. At the same time, much work has been undertaken investigating the efficacy of the small-molecule epidermal growth factor receptor tyrosine kinase inhibitors gefitinib and erlotinib in the second-line and subsequent-line settings. Erlotinib has been shown to significantly improve survival versus placebo following failure on one or two prior regimens, especially in select or targeted patient populations. Finding the optimal place and patients for these agents in NSCLC treatment will be one of the more fascinating challenges of the next few years. PMID- 16472704 TI - Use of novel second-line targeted therapies in non-small cell lung cancer. AB - Non-small cell lung cancer (NSCLC) is the leading cause of cancer death in the United States and throughout the world. This is largely because more than half of lung cancer cases present as metastatic disease, making local therapy for cure impossible. The last decade has seen significant improvement in first-line treatment of NSCLC, including the use of new chemotherapeutic agents with more effective therapeutic profiles. However, standard cytotoxic regimens are still limited and it remains critical to better understand and develop new treatment options for refractory disease. This has included some new second-line therapeutic approaches and has led to a focus on molecular targeted therapy, including agents that block the epidermal growth factor receptor (EGFR) or angiogenesis. EGFR-targeted agents such as gefitinib, erlotinib, and cetuximab have been successfully used for NSCLC treatment, with studies reporting overall response rates of 18.4%, 8.9%, and 3.3%, respectively. Angiogenesis inhibitors such as bevacizumab and ZD6474 have also improved treatment outcome. Bevacizumab had an overall response rate of 27% when used in combination with paclitaxel and carboplatin, and ZD6474 had an overall response rate of 26% when used in combination with docetaxel. Using these compounds alone or in combination may improve the survival and quality of life of patients with lung cancer in the refractory setting. PMID- 16472705 TI - Second-line treatment options in non-small cell lung cancer: a comparison of cytotoxic agents and targeted therapies. AB - Current options for the second-line treatment of non-small cell lung cancer (NSCLC) include cytotoxic drugs, such as docetaxel and pemetrexed, and targeted therapies. Docetaxel was approved in the United States and Europe in 2000 after two phase III trials showed drug superiority versus best supportive care alone and versus alternative single-agent chemotherapy. Pemetrexed was approved in the United States and Europe in 2004 after a phase III trial showed that, compared with docetaxel, it had comparable activity (median survival time of approximately 8 months in both arms) and a more favorable toxicity profile: grade 3-4 neutropenia was observed in 5.3% versus 40.2% of patients in the pemetrexed and docetaxel arms, respectively, while febrile neutropenia was observed in 1.9% versus 12.7% of patients, respectively. In the United States, gefitinib and erlotinib have also been approved for the treatment of recurrent NSCLC (in 2003 and 2004, respectively), while in Europe the registration of these agents is currently under evaluation. This review focuses on the use of docetaxel and pemetrexed for the second-line treatment of NSCLC and compares these drugs with targeted therapies, highlighting the latest developments in pharmacogenomics that might lead to a more tailored approach to treatment. PMID- 16472706 TI - Optimizing therapy in previously treated non-small cell lung cancer. AB - The past decade has seen the identification of several novel cytotoxic agents. More recently, targeted therapies with single-agent activity or that enhance the efficacy of chemotherapy in advanced non-small cell lung cancer have been identified. These agents have been, or are undergoing, active testing alone and in combination in both loco-regional and advanced disease as initial therapy and after failure of cytotoxic therapy. The cytotoxic agents docetaxel, pemetrexed, and topotecan, as well as the epidermal growth factor receptor tyrosine kinase inhibitors erlotinib and gefitinib, have been evaluated in phase III trials in previously treated populations. This review summarizes the results of these phase III studies with a particular focus on predictors of favorable outcome, attempts to provide a rational approach to therapeutic selection in this patient population, and discusses ongoing pivotal trials and future strategies in this field. PMID- 16472707 TI - The emergence of a unique population in non-small cell lung cancer: systemic or loco-regional relapse following postoperative adjuvant platinum-based chemotherapy. AB - Surgical resection remains the foundation of curative therapy in early stage non small cell lung cancer (NSCLC). Recent evidence from several randomized trials has shown that adjuvant chemotherapy regimens of platinum-based agents plus third generation cytotoxic agents improve survival rates in this population of patients. Despite the use of adjuvant chemotherapy, many patients will suffer a relapse, typically systemic, and be candidates for subsequent therapies. This represents a new population of patients with NSCLC. To date, there are no published trials that evaluate prognostic factors or therapeutic outcomes in these patients. Although therapeutic paradigms have been established in patients who present with de novo metastatic NSCLC, it is not clear whether these paradigms are successful for patients who relapse following platinum-based adjuvant chemotherapy. The influence of time from the previous adjuvant exposure is likely to be as important as it seems to be in other solid tumors. However, dedicated clinical research has not yet established this paradigm in NSCLC and the influence this should have on the choice of subsequent therapy is unknown. The nature of the prior adjuvant exposure may also play a role in the choice of subsequent therapy. This has implications for the design of future trials in advanced NSCLC as these patients may or may not have the same natural history as the de novo metastatic NSCLC patient. Future clinical trials are required to address these issues as well as the biologic heterogeneity that likely exists between patients who are seemingly cured by surgery, relapse early (<1 year), or at later times following adjuvant therapy. PMID- 16472708 TI - Geographic variation in the second-line treatment of non-small cell lung cancer. AB - Although there is broad agreement on management options for treating different stages of non-small cell lung cancer (ie, surgery for stage I and II disease, combined treatment modalities for stage III disease, and platinum-based chemotherapy as initial treatment for appropriate patients with stage IV disease), there is considerable geographic variation in practice patterns. These variations reflect a number of factors, including health care economics, the influence of national and regional regulatory bodies, the nature of physician and patient interaction, and probable biological differences between different populations in terms of drug metabolism and inherent susceptibility to both drug activity and toxicity. The approaches taken by three different geographic regions, the United States, European Union, and Japan, are evaluated. Clinically, the most striking differences in activity and toxicity between different regions have been seen with the epidermal growth factor receptor inhibitors gefitinib and erlotinib. Japanese patients experience significantly greater response and a greater degree of interstitial lung disease than patients in the European Union and North America (ie, US and Canada). Similar differences in efficacy and toxicity have also been noted with cytotoxic chemotherapy agents in the first line setting. These geographic and ethnic differences in toxicity and efficacy will need to be considered in the design and comparison of future clinical trials. PMID- 16472709 TI - Future directions in the second-line treatment of non-small cell lung cancer. AB - Single-agent chemotherapy has shown limited activity as second-line treatment in metastatic non-small cell lung cancer (NSCLC), with short-lived responses and modest survival benefit over best supportive care or placebo. There are multiple ways to improve the poor outcome of patients whose disease progresses after first line chemotherapy. First, individualizing second-line chemotherapy could optimize its effect; the discovery of dramatic responses and significant improvement in survival in patients with epidermal growth factor receptor (EGFR) gene mutations who are treated with EGFR tyrosine kinase inhibitors may lead to the application of other novel therapeutic approaches. Cancer vaccines, using autologous tumor cells genetically modified with granulocyte-macrophage colony-stimulating factor, constitute a new therapeutic option for patients with chemoresistant advanced NSCLC. Vaccines based on lymphocyte-defined tumor antigens, such as melanoma associated antigen-3, toll-like receptor 9, and mucin 1, are also in the first stages of testing and have shown promising preliminary results. New approaches in gene therapy, including a p53-based method, are currently being investigated. The ultimate goal of gene therapy is to target cancerous stem cells, the importance of which is beginning to be recognized in NSCLC through the study of abnormalities in the wingless (Wnt) pathway. At the preclinical level, small interfering RNA sequences have been used successfully to neutralize multiple abnormal components of the Wnt pathway. PMID- 16472711 TI - Goals of treatment for patients with metastatic breast cancer. AB - The key goal in the treatment of metastatic breast cancer is to prolong survival, with an emphasis on restricting treatment-related toxicity as much as possible. Despite the plethora of treatment modalities available in metastatic breast cancer, significant survival differences are relatively uncommon. Symptom relief and quality of life are other important, clinically validated measurement instruments. Symptom relief in particular is not used as widely used as it could be, in contrast to lung cancer where it has been proven clinically informative. Finally, time to disease progression is an increasingly used primary endpoint in comparing treatments for metastatic breast cancer; this measure includes both patients who achieve an objective response, and those whose disease may be stabilized with treatment. PMID- 16472712 TI - Role of gemcitabine in breast cancer management: an update. AB - Anthracyclines and the taxanes are now used earlier in the course of therapy for metastatic breast cancer, and increasingly as part of adjuvant treatment. Accordingly, novel therapies are warranted, including alternatives to anthracyclines and combination strategies in the setting of disease resistance to or disease progression after anthracycline and taxane therapy. Gemcitabine is part of this strategy. The gemcitabine/paclitaxel combination has emerged as part of a new standard of combination therapy for advanced breast cancer, based on phase III data showing superiority of this combination over paclitaxel alone in the first-line therapy setting. Gemcitabine alone, or in combination with paclitaxel and other agents, is also undergoing evaluation in other settings including the refractory metastatic setting, as part of management of patients with HER2-positive disease, and also as part of adjuvant therapy for breast cancer. PMID- 16472713 TI - Clinical experience with pemetrexed in breast cancer. AB - Alimta (pemetrexed) is a novel multitargeted antifolate that inhibits several enzymes in the de novo pathways of pyrimidine and purine biosynthesis, including thymidylate synthase, dihydrofolate reductase, and glycinamide ribonucleotide formyltransferase. Pemetrexed possesses antitumor activity in several solid tumors, including non-small cell lung cancer, malignant pleural mesothelioma, pancreas, colorectal, gastric, bladder, breast, and head and neck cancers. The main toxicities of the drug are myelosuppression, skin rash, and mucositis. Both myelosuppression and mucositis are more frequent in patients with high homocysteine plasma levels (an indicator of deficient vitamin B(12) and folate pools). Supplementation with vitamin B(12) and folic acid greatly reduces most severe toxicities and has been implemented in pemetrexed trials since December 1999. Pemetrexed has been tested in five phase II trials in locally advanced or metastatic breast cancer. The drug has shown an activity of around 30% in advanced breast cancer patients with minimal or no prior chemotherapy. In patients who received prior anthracyclines, response rates of 21% were reported. Responses have also been observed in a moderate proportion of patients who had been pretreated with anthracyclines, taxanes, and capecitabine. Some studies have suggested that a correlation exists between thymidylate synthase tumor expression with pemetrexed antitumor activity; this attractive hypothesis should be confirmed in further studies. The optimal dose when combined with vitamin supplementation is under current investigation in patients with breast cancer. A randomized phase II study comparing pemetrexed 600 and 900 mg/m(2) with vitamin supplementation as first-line treatment for metastatic breast cancer is ongoing. PMID- 16472714 TI - Differences in IL-10 and IL-12 production patterns and differences in the effects of acute ethanol treatment on macrophages in vivo and in vitro. AB - Several recent studies have documented that signaling can be fundamentally different in vivo and in vitro. However, studies of signaling and cytokine production by macrophages are often conducted in vitro, without confirmation in vivo. In addition, the direct effects of drugs and chemicals, including ethanol, on these processes are also often investigated in vitro. The purpose of the present study was to compare production of interleukin-6 (IL-6), IL-10, and IL-12 by macrophages in response to two different ligands for toll-like receptors and the effects of acute ethanol exposure on these responses in vivo and in vitro. The macrophage-like cell line RAW 264.7 is also widely used in cytokine and signaling studies, so these cells were also evaluated in this study. The results indicate that IL-6 production and the effects of Ethanol on IL-6 were similar in vivo and in vitro. In contrast, IL-10 was produced to a much greater extent in vitro than in vivo, and IL-12 was often undetectable in vitro even though it was produced at greater concentrations than IL-10 in vivo. To determine the role of altered secretion of preformed IL-10 as compared to new synthesis, cells were treated in vitro with protein and mRNA synthesis inhibitors. The results suggest that preformed IL-10 is released in vivo, but almost all IL-10 secreted in vitro is newly synthesized. Ethanol suppressed IL-12 and enhanced or had no effect on IL-10 production in vivo, whereas it decreased IL-10 production in vitro. These effects were similar at different times and using different concentrations of toll-like receptor ligands. In general, RAW 264.7 cells responded similarly to peritoneal macrophages in vitro. This suggests that results for cytokine studies and probably signaling studies as well that are conducted in vitro should be interpreted with caution and confirmed in vivo, particularly if they involve IL 10 and IL-12. PMID- 16472715 TI - Ethanol withdrawal and proclivity are inversely related in rats selectively bred for differential saccharin intake. AB - Withdrawal severity and voluntary alcohol consumption are inversely related in rats and mice. The present study demonstrated this empirical relation and extended it in two ways. First, the rats were selectively bred for low (LoS) and high (HiS) saccharin intake, a phenotype that correlates positively with ethanol intake and inversely with emotional reactivity. Withdrawal has not yet been studied in these rats. Second, proclivity to consume ethanol was measured as conditioned preference for an ethanol-paired flavor. After 2 weeks of forced exposure to ethanol and a period of abstinence, LoS rats showed elevated acoustic startle; HiS rats did not (Exp. 1). When ethanol- and no-ethanol solutions were available freely during conditioning, both LoS and HiS rats preferred a flavor paired with 4% ethanol, but only HiS rats preferred a flavor paired with 10% ethanol (Exp. 2A); when exposure to the two solutions was controlled, all groups except LoS males preferred flavors paired with 4% or 10% ethanol (Exp. 2B). Thus, as predicted, withdrawal was more severe in the line with less ethanol proclivity (LoS). These results implicate basic associative and affective processes in individual differences in patterns of alcohol use. PMID- 16472717 TI - Discriminative stimulus effects of 5.6 mg/kg pregnanolone in DBA/2J and C57BL/6J inbred mice. AB - Neurosteroids represent a class of endogenous compounds that exert rapid, nongenomic effects through neurotransmitter receptor systems such as gamma aminobutyric acid(A) (GABA(A)). Two neurosteroids, allopregnanolone (3alpha hydroxy-5alpha-pregnan-20-one) and pregnanolone (3alpha-hydroxy-5beta-pregnan-20 one), possess anxiolytic and sedative properties and show substitution for ethanol, benzodiazepines, and barbiturates in drug discrimination assays. A previous study examining the discriminative stimulus effects of 10 mg/kg pregnanolone in DBA/2J and C57BL/6J mice showed pregnanolone's discriminative stimulus to be mediated primarily through GABA(A) positive modulation. This study examined the discriminative stimulus effects of a lower training dose (5.6 mg/kg) of pregnanolone in DBA/2J and C57BL/5J mice. Twelve male DBA/2J mice and 12 male C57BL/6J mice were trained to discriminate 5.6 mg/kg pregnanolone. GABA(A) receptor positive modulators, neuroactive steroids, NMDA receptor antagonists, and 5-HT(3) receptor agonists were tested for pregnanolone substitution. In DBA/2J and C57BL/6J mice benzodiazepine, barbiturate, and GABAergic neuroactive steroids all substituted for pregnanolone. In the DBA/2J mice, NMDA receptor antagonists showed generalization to the discriminative stimulus cues of pregnanolone, an effect not seen in the C57BL/6J mice. 5-HT(3) receptor agonists and zolpidem failed to substitute for pregnanolone's discriminative stimulus in either strain. AlloTHDOC and midazolam were more potent in producing pregnanolone like discriminative stimulus effects in DBA/2J mice. These results provide a comprehensive look at pregnanolone's discriminative stimulus effects in two commonly used strains of mice. The present data suggest GABA(A)-receptor positive modulation as the predominant receptor mechanism mediating the discriminative stimulus effects of pregnanolone. NMDA receptor antagonism was suggested in the DBA/2J mice and may represent a heterogenous cue produced by the lower training dose of pregnanolone. PMID- 16472718 TI - Saccharin consumption and the effect of a long-term exposure to a sweetened alcoholic solution in high- (UChB) and low- (UChA) alcohol-drinking rats. AB - An association between saccharin consumption and alcohol intake has been observed in rodent lines genetically developed for alcohol preference or alcohol avoidance. It has also been proposed that a sweetened alcohol solution can condition rats to consume high amounts of alcohol. This work had two aims. First, to study the relationship between saccharin and alcohol intake in both high alcohol-drinking UChB rats and low-alcohol-drinking UChA rats and, second, to determine whether a long-term exposure to a sweetened alcohol solution can increase their voluntary alcohol consumption. For the first purpose, UChB and UChA rats were tested under a free-choice paradigm between two graduated bottles, one containing a saccharin solution (0.1, 0.2, or 0.4% [wt/vol]) and the other water. For the second purpose, UChB and UChA rats that were under free choice between 10% alcohol and water, were offered a 10% alcohol solution containing 0.2% saccharin, instead of 10% alcohol for 1 month and were then returned to free choice between 10% alcohol and water. The first experiment showed that both lines have a high preference for saccharin at any concentration, but UChB rats drank twice as much saccharin solution as UChA rats and consequently they increased significantly their total daily fluid intake. This fact has been suggested to be an animal analogue of the clinical phenomenon known as "loss of control." In the second experiment a long-term exposure to a 10% alcohol solution containing 0.2% saccharin induced a significant increase in alcohol consumption in UChB rats once saccharin was faded out, whereas the alcohol consumption in UChA rats returned to the previous low value. This result indicates that UChA rats have a genetic predisposition to avoid alcohol. In conclusion, the results reported here for UChB and UChA rats show an association between saccharin and alcohol preference, and suggest that their different genotypes are probably involved in alcohol aversion. PMID- 16472716 TI - Treatment with and withdrawal from finasteride alter ethanol intake patterns in male C57BL/6J mice: potential role of endogenous neurosteroids? AB - Exogenous administration of the gamma-aminobutyric acid (GABA)-ergic neurosteroid allopregnanolone (ALLO) can increase ethanol intake in rats and mice. To determine the contribution of endogenous neurosteroids (i.e., ALLO and related pregnane steroids) in the regulation of established ethanol consumption patterns in male C57BL/6J (B6) mice, the 5alpha-reductase (5alpha-R) enzyme inhibitor, finasteride (FIN), was chronically administered and then subsequently withdrawn. Mice were provided daily 2-h limited access to a 10% vol/vol ethanol solution (10E) and water in lickometer chambers during the dark phase. Following the establishment of stable 10E intake patterns, mice were injected intraperitoneally with either vehicle (20% wt/vol 2-hydroxypropyl-beta-cyclodextrin; n=8) or FIN (50 mg/kg; n=16) for 7 days. Effects of withdrawal from FIN treatment were subsequently assessed for an additional 7 days. Ethanol intakes were significantly decreased with acute FIN treatment (days 1-3) and during early withdrawal (days 1-3). Acute FIN treatment was also associated with an extended latency to first bout, reduced first bout size, and greatly attenuated sipper contact count during the initial 20-min interval of 10E access. These findings collectively indicated that acute FIN treatment markedly attenuated the initiation of 10E consumption during the limited access sessions. The influence of FIN on 10E intake patterns was largely dissipated with chronic treatment, suggesting that compensatory changes in neurosteroid modulation of inhibitory tone may have occurred. Thus, acute FIN treatment modulated ethanol intake patterns in a manner opposite to that previously demonstrated for a physiologically relevant, exogenous ALLO dose, consistent with the ability of a alpha-R inhibitor to block ALLO biosynthesis. Manipulation of endogenous neurosteroid activity via biosynthetic enzyme inhibition or antagonism of steroid binding to the GABA type A receptor may prove to be a beneficial pharmacotherapeutic strategy in the intervention of alcohol abuse and alcoholism. PMID- 16472719 TI - Early and late effects of alcohol ingestion on blood pressure and endothelial function. AB - Previous investigations have shown a biphasic effect of alcohol on blood pressure (BP). However, there are no studies on possible simultaneous influences in endothelial function. This study aims to evaluate the early and late effects of alcohol ingestion on vascular and endothelial function parameters in healthy young men. The diameter of brachial artery (DBA), endothelium-dependent flow mediated dilatation, endothelium-independent nitroglycerin-mediated dilatation, systolic blood pressure (SBP), diastolic blood pressure (DBP), and heart rate were measured 30 min before intake, 4 h after intervention (when there is a reported hypotensive effect of alcohol), and after 13 h (subsequent increase in BP). The study group consisted of 100 males aged 18-25 years who were evaluated by brachial artery ultrasound. Subjects were randomized to drink either an alcoholic (60 g of ethanol) or a similar nonalcoholic beverage. Alcohol induced a biphasic effect on SBP and DBP, with a 4-h decrease followed by an increase after 13 h. After 4 h, the alcohol-drinking group presented a DBA increase that was significant at baseline and after hyperemia but not after nitroglycerin administration. There were no DBA differences between the intervention and control groups 13 h after drinking. This study replicates the initial reports of alcohol-induced biphasic alteration in BP. Our results showed that despite the late increase in BP, there were no accompanying changes in endothelial function. PMID- 16472720 TI - Brain mechanisms of expectation associated with insula and amygdala response to aversive taste: implications for placebo. AB - The experience of aversion is shaped by multiple physiological and psychological factors including one's expectations. Recent work has shown that expectancy manipulation can alter perceptions of aversive events and concomitant brain activation. Accruing evidence indicates a primary role of altered expectancies in the placebo effect. Here, we probed the mechanism by which expectation attenuates sensory taste transmission by examining how brain areas activated by misleading information during an expectancy period modulate insula and amygdala activation to a highly aversive bitter taste. In a rapid event-related fMRI design, we showed that activations in the rostral anterior cingulate cortex (rACC), orbitofrontal cortex (OFC), and dorsolateral prefrontal cortex to a misleading cue that the taste would be mildly aversive predicted decreases in insula and amygdala activation to the highly aversive taste. OFC and rACC activation to the misleading cue were also associated with less aversive ratings of that taste. Additional analyses revealed consistent results demonstrating functional connectivity among the OFC, rACC, and insula. Altering expectancies of upcoming aversive events are shown here to depend on robust functional associations among brain regions implicated in prior work on the placebo effect. PMID- 16472722 TI - Diseases manifesting in the upper genital tract in children and adolescents: a review. PMID- 16472724 TI - Self-reported physical activity and bone mineral density in urban adolescent girls. AB - BACKGROUND: This observational study aimed to examine the prevalence of activities of daily living, as well as the impact of leisure time activities, on bone mineral density in urban adolescent girls. METHODS: Patients completed a 23 item physical activity questionnaire at baseline, recording time spent in various activities in the previous 7 days. In addition to leisure time activities, activities of daily life were also considered. Activities were characterized and scored by metabolic intensity (METPA) and mechanical strain on bone (MECHPA). The METPA score for each activity is the product of the metabolic intensity of the activity and the time spent in the activity. The MECHPA score estimates the mechanical strain on bone from ground reaction forces. The logged scores were divided into quartiles with the lowest quartile as the reference group. RESULTS: Four hundred fifty-five females (ages 12-18 years) completed the survey (62% black and 38% non-black). The log of the overall METPA score was a significant predictor of bone mineral density (i.e. higher METPA score predicted a higher bone mineral density, P = 0.03). A MECHPA score in the highest quartile was associated with a higher bone mineral density (P < 0.05) when compared to the other MECHPA quartiles. CONCLUSIONS: In this population of urban adolescent girls, activities of daily living were reported with a higher frequency than sports activities. Results indicated a positive association between the time spent in metabolically intense activities and bone mineral density. There also appears to be a threshold effect for the relationship between activities with the highest mechanical strain and bone mineral density. PMID- 16472725 TI - Effect of specimen order on Chlamydia trachomatis and Neisseria gonorrhoeae test performance and adequacy of Papanicolaou smear. AB - STUDY OBJECTIVE: To evaluate the effect of specimen collection order on the performance of diagnostic tests for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC), and the specimen adequacy of ThinPrep Papanicolaou (Pap) smears. DESIGN: Prospective cohort study. SETTING: Public adolescent clinic. POPULATION: 313 women. INTERVENTIONS: The order of five cervical testing specimens was randomized for (1) ligase chain reaction (LCR) and (2) polymerase chain reaction (PCR) for both CT and GC; (3) transcription-mediated amplification (TMA) for CT; (4) culture for CT; and (5) cytology and LCR for GC and CT performed on ThinPrep Pap specimens. For CT and GC, a reference standard was based on three different tests performed on separate specimens. Generalized estimating equations were used to account for repeated measures. MAIN OUTCOME MEASURE: Sensitivity and specificity of diagnostic tests. RESULTS: The proportion of inadequate Pap smears was independent of specimen order. As a group, nucleic acid amplification test (NAAT) sensitivity and specificity for GC and CT were similar in the first two (early) and last three (late) swabs. Although point estimates for sensitivity were higher in the early swabs compared to the late swabs for GC LCR (13% difference), GC PCR (13%), and CT TMA (10%), these differences were not statistically significant. Their clinical significance warrants further investigation. CONCLUSIONS: In clinical settings where both Pap smears and STI testing are performed in adolescents, clinical considerations can influence the order of specimen collection, since neither Pap specimen adequacy nor test performance of NAAT for CT and GC were significantly associated with swab order. PMID- 16472726 TI - Effect of perineal cleansing on contamination rate of mid-stream urine culture. AB - STUDY OBJECTIVE: Urinary tract infection (UTI) and chlamydial or gonococcal urethritis are the most common causes of female dysuria. While chlamydia and gonorrhea can be detected with a nucleic acid amplification test performed on an uncleansed first part voided urine sample, urine cultures to test for UTI require a mid-stream clean caught sample. In order to determine whether collecting two sequential non-clean caught urine samples during the same void to test for chlamydia, gonorrhea, and UTI is a reasonable approach, we assessed the degree to which perineal cleansing reduces bacterial contamination of mid-stream urine cultures. DESIGN: Experimental study comparing mid-stream urine samples collected with (n = 25) and without (n = 25) perineal cleansing. SETTING: A university based adolescent clinic PARTICIPANTS: We recruited fifty 14-23-year-old (mean 18.5 yrs, SD 2.3) asymptomatic females. MAIN OUTCOME MEASURE: Perineal flora contamination rate of mid-stream urine cultures collected with and without perineal cleansing. RESULTS: No culture grew >10(4) colonies of a pathogenic bacterium. Eleven (44%) of the experimental group samples and 9 (36%) of the control samples grew >10(4) colonies of perineal bacterial flora (chi(2) = .33, P = 0.56). Participants' previous experience collecting mid-stream urine was not associated with less bacterial contamination. CONCLUSIONS: Among this small sample of asymptomatic young women, perineal cleansing did not significantly reduce perineal flora contamination of mid-stream urine cultures. If larger studies of symptomatic young women replicate these findings, young women could collect two sequential urine samples from the same void to test for chlamydia, gonorrhea, and UTI. PMID- 16472727 TI - Rapidly growing bilateral ovarian cystadenoma in a 6-year-old girl: case report and literature review. AB - BACKGROUND: Benign ovarian neoplasms originating from epithelial tissue are common tumors in adult women. However, they are rarely seen in the pediatric population, especially in the first decade of life. CASE: We report a case of a 6 year-old, premenarchal girl, previously healthy, with frequent micturition lasting 3 days prior to the first examination, without discomfort or pain. Laboratory analyses of blood and urine showed no abnormalities. Repeated ultrasonographic examinations revealed bilateral, cystic, rapidly growing ovarian masses. Cysts were surgically removed, with preservation of normal ovarian tissue, and histopathologic findings showed a serous cystadenoma of both ovaries. PMID- 16472728 TI - Anaplastic carcinoma arising in an ovarian mucinous cystadenocarcinoma in a 17 year-old female. AB - BACKGROUND: Anaplastic carcinoma arising within a mucinous ovarian neoplasm is rare, with only about 30 reported cases. Reported cases have given a broad age range, ranging from 17 to 72 years of age, but occurrence in adolescents is exceptional, with only a few cases reported. CASE: We report a case of anaplastic carcinoma arising in a mucinous cystadenocarcinoma in a 17-year-old female who presented with severe abdominal pain, an unusual symptom for an ovarian malignancy in the postmenopausal patient, but not in the adolescent. The patient had widespread metastases at the time of presentation, consistent with the aggressive behavior of this neoplasm. CONCLUSIONS: This case illustrates that, although rare, epithelial ovarian malignancy is in the differential diagnosis of abdominal pain in an adolescent. PMID- 16472729 TI - What's your diagnosis? Painful nodules on the perineum with scarring and sinus tract formation. PMID- 16472730 TI - Dysfunctional uterine bleeding in adolescents. PMID- 16472731 TI - Management quandary. Menstrual issues in a teenager with developmental delay. PMID- 16472732 TI - Home visiting for the pregnant and parenting teen. PMID- 16472733 TI - The most complex pseudouridylase. PMID- 16472734 TI - A lock on formins. PMID- 16472735 TI - Selectivity and promiscuity in Eph receptors. PMID- 16472736 TI - Decapper comes into focus. PMID- 16472737 TI - Structure of the catalytic and ubiquitin-associated domains of the protein kinase MARK/Par-1. AB - The Ser/Thr kinase MARK2 phosphorylates tau protein at sites that cause detachment from microtubules in Alzheimer neurofibrillary degeneration. Homologs of MARK2 include Par-1 in C. elegans and Drosophila, which generates embryonic polarity. We report the X-ray structure of the catalytic and ubiquitin-associated domains (UBA) of human MARK2. The activity was altered by mutations in the ATP binding site and/or activation loop. The catalytic domain shows the small and large lobes typical of kinases. The substrate cleft is in an inactive, open conformation in the inactivated and the wild-type structure. The UBA domain is attached via a taut linker to the large lobe of the kinase domain and leans against a hydrophobic patch on the small lobe. The UBA structure is unusual because the orientation of its third helix is inverted, relative to previous structures. Possible implications of the structure for the regulation of kinase activity are discussed. PMID- 16472738 TI - The crystal structure of Aspergillus fumigatus cyclophilin reveals 3D domain swapping of a central element. AB - The crystal structure of Aspergillus fumigatus cyclophilin (Asp f 11) was solved by the multiwavelength anomalous dispersion method and was refined to a resolution of 1.85 A with R and R(free) values of 18.9% and 21.4%, respectively. Many cyclophilin structures have been solved to date, all showing the same monomeric conformation. In contrast, the structure of A. fumigatus cyclophilin reveals dimerization by 3D domain swapping and represents one of the first proteins with a swapped central domain. The domain-swapped element consists of two beta strands and a subsequent loop carrying a conserved tryptophan. The tryptophan binds into the active site, inactivating cis-trans isomerization. This might be a means of biological regulation. The two hinge loops leave the protein prone to misfolding. In this context, alternative forms of 3D domain swapping that can lead to N- or C-terminally swapped dimers, oligomers, and aggregates are discussed. PMID- 16472739 TI - Modeling a self-avoiding chromatin loop: relation to the packing problem, action at-a-distance, and nuclear context. AB - There is now convincing evidence that genomes are organized into loops, and that looping brings distant genes together so that they can bind to local concentrations of polymerases in "factories" or "hubs." As there remains no systematic analysis of how looping affects the probability that a gene can access binding sites in such factories/hubs, we used an algorithm that we devised and Monte Carlo methods to model a DNA or chromatin loop as a semiflexible (self avoiding) tube attached to a sphere; we examine how loop thickness, rigidity, and contour length affect where particular segments of the loop lie relative to binding sites on the sphere. Results are compared with those obtained with the traditional model of an (infinitely thin) freely jointed chain. They provide insights into the packing problem (how long genomes are packed into small nuclei), and action-at-a-distance (how firing of one origin or gene can prevent firing of an adjacent one). PMID- 16472740 TI - Complete reaction cycle of a cocaine catalytic antibody at atomic resolution. AB - Antibody 7A1 hydrolyzes cocaine to produce nonpsychoactive metabolites ecgonine methyl ester and benzoic acid. Crystal structures of 7A1 Fab' and six complexes with substrate cocaine, the transition state analog, products ecgonine methyl ester and benzoic acid together and individually, as well as heptaethylene glycol have been analyzed at 1.5-2.3 angstroms resolution. Here, we present snapshots of the complete cycle of the cocaine hydrolytic reaction at atomic resolution. Significant structural rearrangements occur along the reaction pathway, but they are generally limited to the binding site, including the ligands themselves. Several interacting side chains either change their rotamers or alter their mobility to accommodate the different reaction steps. CDR loop movements (up to 2.3 angstroms) and substantial side chain rearrangements (up to 9 angstroms) alter the shape and size (approximately 320-500 angstroms3) of the antibody active site from "open" to "closed" to "open" for the substrate, transition state, and product states, respectively. PMID- 16472741 TI - Flexibility of thiamine diphosphate revealed by kinetic crystallographic studies of the reaction of pyruvate-ferredoxin oxidoreductase with pyruvate. AB - Pyruvate-ferredoxin oxidoreductases (PFOR) are unique among thiamine pyrophosphate (ThDP)-containing enzymes in giving rise to a rather stable cofactor-based free-radical species upon the decarboxylation of their first substrate, pyruvate. We have obtained snapshots of unreacted and partially reacted (probably as a tetrahedral intermediate) pyruvate-PFOR complexes at different time intervals. We conclude that pyruvate decarboxylation involves very limited substrate-to-product movements but a significant displacement of the thiazolium moiety of ThDP. In this respect, PFOR seems to differ substantially from other ThDP-containing enzymes, such as transketolase and pyruvate decarboxylase. In addition, exposure of PFOR to oxygen in the presence of pyruvate results in significant inhibition of catalytic activity, both in solution and in the crystals. Examination of the crystal structure of inhibited PFOR suggests that the loss of activity results from oxime formation at the 4' amino substituent of the pyrimidine moiety of ThDP. PMID- 16472742 TI - Crystal structure of group A streptococcus Mac-1: insight into dimer-mediated specificity for recognition of human IgG. AB - Group A Streptococcus secretes cysteine proteases named Mac-1 and Mac-2 that mediate host immune evasion by targeting both IgG and Fc receptors. Here, we report the crystal structures of Mac-1 and its catalytically inactive C94A mutant in two different crystal forms. Despite the lack of sequence homology, Mac-1 adopts the canonical papain fold. Alanine mutations at the active site confirmed the critical residues involved in a papain-like catalytic mechanism. Mac-1 forms a symmetric dimer in both crystal forms and displays the unique dimer interface among papain superfamily members. Mutations at the dimer interface resulted in a significant reduction in IgG binding and catalysis, suggesting that the dimer contributes to both IgG specificity and enzyme cooperativity. A tunnel observed at the dimer interface constitutes a target for designing potential Mac-1 specific antimicrobial agents. The structures also offer insight into the functional difference between Mac-1 and Mac-2. PMID- 16472743 TI - Ligand-induced domain rearrangement of fatty acid beta-oxidation multienzyme complex. AB - The quaternary structure of a fatty acid beta-oxidation multienzyme complex, catalyzing three sequential reactions, was investigated by X-ray crystallographic and small-angle X-ray solution scattering analyses. X-ray crystallography revealed an intermediate structure of the complex among the previously reported structures. However, the theoretical scattering curves calculated from the crystal structures remarkably disagree with the experimental profiles. Instead, an ensemble of the atomic models, which were all calculated by rigid-body optimization, reasonably explained the experimental data. These structures significantly differ from those in the crystals, but they maintain the substrate binding pocket at the domain boundary. Comparisons among these structures indicated that binding of 3-hydroxyhexadecanoyl-CoA or nicotinamide adenine dinucleotide induces domain rearrangements in the complex. The conformational changes suggest the structural events occurring during the chain reaction catalyzed by the multienzyme complex. PMID- 16472744 TI - Antiparallel four-stranded coiled coil specified by a 3-3-1 hydrophobic heptad repeat. AB - Coiled-coil sequences in proteins commonly share a seven-amino acid repeat with nonpolar side chains at the first (a) and fourth (d) positions. We investigate here the role of a 3-3-1 hydrophobic repeat containing nonpolar amino acids at the a, d, and g positions in determining the structures of coiled coils using mutants of the GCN4 leucine zipper dimerization domain. When three charged residues at the g positions in the parental sequence are replaced by nonpolar alanine or valine side chains, stable four-helix structures result. The X-ray crystal structures of the tetramers reveal antiparallel, four-stranded coiled coils in which the a, d, and g side chains interlock in a combination of knobs into-knobs and knobs-into-holes packing. Interfacial interactions in a coiled coil can therefore be prescribed by hydrophobic-polar patterns beyond the canonical 3-4 heptad repeat. The results suggest that the conserved, charged residues at the g positions in the GCN4 leucine zipper can impart a negative design element to disfavor thermodynamically more stable, antiparallel tetramers. PMID- 16472746 TI - The backrub motion: how protein backbone shrugs when a sidechain dances. AB - Surprisingly, the frozen structures from ultra-high-resolution protein crystallography reveal a prevalent, but subtle, mode of local backbone motion coupled to much larger, two-state changes of sidechain conformation. This "backrub" motion provides an influential and common type of local plasticity in protein backbone. Concerted reorientation of two adjacent peptides swings the central sidechain perpendicular to the chain direction, changing accessible sidechain conformations while leaving flanking structure undisturbed. Alternate conformations in sub-1 angstroms crystal structures show backrub motions for two thirds of the significant Cbeta shifts and 3% of the total residues in these proteins (126/3882), accompanied by two-state changes in sidechain rotamer. The Backrub modeling tool is effective in crystallographic rebuilding. For homology modeling or protein redesign, backrubs can provide realistic, small perturbations to rigid backbones. For large sidechain changes in protein dynamics or for single mutations, backrubs allow backbone accommodation while maintaining H bonds and ideal geometry. PMID- 16472745 TI - Structure of the autoinhibitory switch in formin mDia1. AB - Diaphanous-related formins (DRFs) regulate the nucleation and polymerization of unbranched actin filaments. The activity of DRFs is inhibited by an intramolecular interaction between their N-terminal regulatory region and a conserved C-terminal segment termed the Diaphanous autoinhibitory domain (DAD). Binding of GTP bound Rho to the mDia1 N terminus releases this autoinhibitory restraint. Here, we describe the crystal structure of the DAD segment of mDia1 in complex with the relevant N-terminal fragment, termed the DID domain. The structure reveals that the DAD segment forms an amphipathic helix that binds a conserved, concave surface on the DID domain. Comparison with the structure of the mDia1 N terminus bound to RhoC suggests that release of the autoinhibitory DAD interaction is accomplished largely by Rho-induced restructuring of the adjacent GTPase binding subdomain (GBD), but also by electrostatic repulsion and a small, direct steric occlusion of the DAD binding cleft by Rho itself. PMID- 16472747 TI - Structural and functional aspects of the sensor histidine kinase PrrB from Mycobacterium tuberculosis. AB - We describe the solution structures of two- and three-domain constructs of the sensor histidine kinase PrrB from Mycobacterium tuberculosis, which allow us to locate the HAMP linker relative to the ATP binding and dimerization domains. We show that the three-domain construct is active both for autophosphorylation and for phosphotransfer to the cognate response regulator, PrrA. We also describe the high-resolution crystal structure of the catalytic domain alone, and we show that, in solution, it binds ATP. The conformational flexibility of this domain is discussed and related to other structural information. PMID- 16472748 TI - A versatile conformational switch regulates reactivity in human branched-chain alpha-ketoacid dehydrogenase. AB - The dehydrogenase/decarboxylase (E1b) component of the 4 MD human branched-chain alpha-ketoacid dehydrogenase complex (BCKDC) is a thiamin diphosphate (ThDP) dependent enzyme. We have determined the crystal structures of E1b with ThDP bound intermediates after decarboxylation of alpha-ketoacids. We show that a key tyrosine residue in the E1b active site functions as a conformational switch to reduce the reactivity of the ThDP cofactor through interactions with its thiazolium ring. The intermediates do not assume the often-postulated enamine state, but likely a carbanion state. The carbanion presumably facilitates the second E1b-catalyzed reaction, involving the transfer of an acyl moiety from the intermediate to a lipoic acid prosthetic group in the transacylase (E2b) component of the BCKDC. The tyrosine switch further remodels an E1b loop region to promote E1b binding to E2b. Our results illustrate the versatility of the tyrosine switch in coordinating the catalytic events in E1b by modulating the reactivity of reaction intermediates. PMID- 16472749 TI - Crystal structure of the cytoplasmic domain of the chloride channel ClC-0. AB - Ion channels are frequently organized in a modular fashion and consist of a membrane-embedded pore domain and a soluble regulatory domain. A similar organization is found for the ClC family of Cl- channels and transporters. Here, we describe the crystal structure of the cytoplasmic domain of ClC-0, the voltage dependent Cl- channel from T. marmorata. The structure contains a folded core of two tightly interacting cystathionine beta-synthetase (CBS) subdomains. The two subdomains are connected by a 96 residue mobile linker that is disordered in the crystals. As revealed by analytical ultracentrifugation, the domains form dimers, thereby most likely extending the 2-fold symmetry of the transmembrane pore. The structure provides insight into the organization of the cytoplasmic domains within the ClC family and establishes a framework for guiding future investigations on regulatory mechanisms. PMID- 16472750 TI - Molecular basis for phosphorylation-dependent, PEST-mediated protein turnover. AB - Proteasomal-mediated rapid turnover of proteins is often modulated by phosphorylation of PEST sequences. The E2 protein from papillomavirus participates in gene transcription, DNA replication, and episomal genome maintenance. Phosphorylation of a PEST sequence located in a flexible region accelerates its degradation. NMR analysis of a 29 amino acid peptide fragment derived from this region shows pH-dependent polyproline II and alpha helix structures, connected by a turn. Phosphorylation, in particular that at serine 301, disrupts the overall structure, and point mutations have either stabilizing or destabilizing effects. There is an excellent correlation between the thermodynamic stability of different peptides and the half-life of E2 proteins containing the same mutations in vivo. The structure around the PEST region appears to have evolved a marginal stability that is finely tunable by phosphorylation. Thus, conformational stability, rather than recognition of a phosphate modification, modulates the degradation of this PEST sequence by the proteasome machinery. PMID- 16472752 TI - Crystal structures of U8 snoRNA decapping nudix hydrolase, X29, and its metal and cap complexes. AB - X29, a 25 kDa Nudix hydrolase from Xenopus laevis that cleaves 5' caps from U8 snoRNA, crystallizes as a homodimeric apoenzyme. Manganese binds crystals of apo X29 to form holo-X29 only in the presence of nucleot(s)ide. Structural changes in X29 on nucleo-t(s)ide-assisted Mn(+2) uptake account for the observed cooperativity of metal binding. Structures of X29 with GTP or m7GpppA show a different mode of ligand binding from that of other cap binding proteins and suggest a possible three- or four-metal Nudix reaction mechanism. The X29 dimer has no known RNA binding motif, but its striking surface dipolarity and unique structural features create a plausible RNA binding channel on the positive face of the protein. Because U8 snoRNP is essential for accumulation of mature 5.8S and 28S rRNA in vertebrate ribosome biogenesis, and cap structures are required for U8 stability in vivo, X29 could profoundly influence this fundamental cellular pathway. PMID- 16472751 TI - Structure and thermodynamic characterization of the EphB4/Ephrin-B2 antagonist peptide complex reveals the determinants for receptor specificity. AB - The Eph receptor tyrosine kinases and their ligands, the ephrins, regulate numerous biological processes in developing and adult tissues and have been implicated in cancer progression and in pathological forms of angiogenesis. We report the crystal structure of the EphB4 receptor in complex with a highly specific antagonistic peptide at a resolution of 1.65 angstroms. The peptide is situated in a hydrophobic cleft of EphB4 corresponding to the cleft in EphB2 occupied by the ephrin-B2 G-H loop, consistent with its antagonistic properties. Structural analysis identifies several residues within the EphB4 binding cleft that likely determine the ligand specificity of this receptor, while isothermal titration calorimetry experiments with truncated forms of the peptide define the amino acid residues of the peptide that are critical for receptor binding. These studies reveal structural features that will aid drug discovery initiatives to develop EphB4 antagonists for therapeutic applications. PMID- 16472754 TI - Structural basis for sulfur relay to RNA mediated by heterohexameric TusBCD complex. AB - Uridine at wobble position 34 of tRNA(Lys), tRNA(Glu), and tRNA(Gln) is exclusively modified into 2-thiouridine (s2U), which is crucial for both precise codon recognition and recognition by the cognate aminoacyl-tRNA synthetases. Recent Escherichia coli genetic studies revealed that the products of five novel genes, tusABCDE, function in the s2U modification. Here, we solved the 2.15 angstroms crystal structure of the E. coli TusBCD complex, a sulfur transfer mediator, forming a heterohexamer composed of a dimer of the heterotrimer. Structure-based sequence alignment suggested two putative active site Cys residues, Cys79 (in TusC) and Cys78 (in TusD), which are exposed on the hexameric complex. In vivo mutant analyses revealed that only Cys78, in the TusD subunit, participates in sulfur transfer during the s2U modification process. Since the single Cys acts as a catalytic residue, we proposed that TusBCD mediates sulfur relay via a putative persulfide state of the TusD subunit. PMID- 16472753 TI - Structure and specific RNA binding of ADAR2 double-stranded RNA binding motifs. AB - Adenosine deaminases that act on RNA (ADARs) site-selectively modify adenosines to inosines within RNA transcripts, thereby recoding genomic information. How ADARs select specific adenosine moieties for deamination is poorly understood. Here, we report NMR structures of the two double-stranded RNA binding motifs (dsRBMs) of rat ADAR2 and an NMR chemical shift perturbation study of the interaction of the two dsRBMs with a 71 nucleotide RNA encoding the R/G site of the GluR-B. We have identified the protein and the RNA surfaces involved in complex formation, allowing us to present an NMR-based model of the complex. We have found that dsRBM1 recognizes a conserved pentaloop, whereas dsRBM2 recognizes two bulged bases adjacent to the editing site, demonstrating RNA structure-dependent recognition by the ADAR2 dsRBMs. In vitro mutagenesis studies with both the protein and the RNA further support our structural findings. PMID- 16472755 TI - Crystal structure of the human retinitis pigmentosa 2 protein and its interaction with Arl3. AB - The crystal structure of human retinitis pigmentosa 2 protein (RP2) was solved to 2.1 angstroms resolution. It consists of an N-terminal beta helix and a C terminal ferredoxin-like alpha/beta domain. RP2 is functionally and structurally related to the tubulin-specific chaperone cofactor C. Seven of nine known RP2 missense mutations identified in patients are located in the beta helix domain, and most of them cluster to the hydrophobic core and are likely to destabilize the protein. Two residues, Glu138 and the catalytically important Arg118, are solvent-exposed and form a salt bridge, indicating that Glu138 might be critical for positioning Arg118 for catalysis. RP2 is a specific effector protein of Arl3. The N-terminal 34 residues and beta helix domain of RP2 are required for this interaction. The abilitities of RP2 to bind Arl3 and cause retinitis pigmentosa seem to be correlated, since both the R118H and E138G mutants show a drastically reduced affinity to Arl3. PMID- 16472756 TI - A phosphorylation-induced conformation change in dematin headpiece. AB - Dematin is an actin binding protein from the junctional complex of the erythrocyte cytoskeleton. The protein has two actin binding sites and bundles actin filaments in vitro. This actin bundling activity is reversibly regulated by phosphorylation in the carboxyl terminal "headpiece" domain (DHP). DHP is a typical villin-type headpiece actin binding motif and contains a flexible N terminal loop and an alpha-helical C-terminal subdomain that is phosphorylated at Ser74. The NMR structure of a Ser74-to-Glu mutant (DHPs74e) closely mimics the conformation of phosphorylated DHP. The negative charge at Ser74 does not alter the conformation of the C-terminal subdomain, but attracts the N-terminal loop toward the C terminus, changing the orientation of the N-terminal subdomain. NMR relaxation studies also indicate reduced mobility in the N-terminal loop in DHPs74e. Thus, phosphorylation in DHP serves as a switch controlling the conformational state of DHP and the actin bundling activity of dematin. PMID- 16472757 TI - Development of high-throughput spermidine synthase activity assay using homogeneous time-resolved fluorescence. AB - Spermidine synthase (SPDS) catalyzes transfer of the propylamine group from decarboxylated S-adenosylmethionine (dcSAM) to putrescine to yield methylthioadenosine (MTA) and spermidine. SPDS plays a regulatory role in cell proliferation and differentiation. This article describes the development of a high-throughput SPDS activity assay using homogeneous time-resolved fluorescence (HTRF) based on energy transfer from europium cryptate as a donor to crosslinked allophycocyanin (XL665) as an acceptor. First a highly specific anti-MTA monoclonal antibody, MTA-7H8, was generated, and then a competitive immunoassay for MTA determination was developed using europium cryptate-labeled MTA-7H8 and XL665-labeled MTA. In our homogeneous immunoassay, the percentage molar cross reactivity of dcSAM with MTA-7H8 was 0.01% and the detection limit of MTA was 2.6 pmol/well. Our HTRF assay uses only one assay plate in which both enzyme reaction and MTA determination can be done successively. Therefore, our method can enable automatic screening of SPDS inhibitors from large numbers of samples. PMID- 16472758 TI - Detection of oxidative stress-induced mitochondrial DNA damage using fluorescence correlation spectroscopy. AB - Using fluorescence correlation spectroscopy (FCS), we tested the feasibility of rapid detection of oxidative damage of mitochondrial DNA (mtDNA) in a small volume. The complete mtDNA genome was amplified by long polymerase chain reaction (LPCR), and the product was fluorescently labeled with an intercalating dye, YOYO 1. The fluorescence autocorrelation function was analyzed using a simple two component model with the diffusion time of 0.21 ms for the LPCR primer and 18 ms for the mtDNA LPCR product. When human embryonic kidney 293 (HEK-293) cells were exposed to 0.4 mM H2O2, the fraction of the mtDNA LPCR product decreased significantly. In contrast, the fraction of the nuclear-encoded beta-globin LPCR product remained unchanged. The analysis time of FCS measurement was very short (5 min) compared with that of gel electrophoresis (3 h). Thus, FCS allowed the rapid detection of the vulnerability of mtDNA to oxidative stress within a small volume element at the subfemtoliter level in solution. These results suggest that the LPCR-FCS method can be used for epidemiological studies of diseases caused by mtDNA damage. PMID- 16472759 TI - [Extragastrointestinal stromal tumors: a report of 4 cases]. AB - Gastrointestinal stromal tumors (GIST) are mesenchymal tumors that arise from the wall of the gastrointestinal tract expressing CD117 and/or the CD34. Similar tumors were described in the soft tissue of the abdomen and are so-called extragastrointestinal stromal tumors (EGIST). We report 4 new cases of EGIST. The tumors occurred in 2 women and 2 men, who ranged in age from 42 to 71 years. Three tumors arose from the soft tissue of the abdominal cavity, and the remainder arose from the retroperitoneum. They ranged in size from 10 to 27 cm. Three cases were composed purely of short fusiform cells tumors, the last case showed an epithelioid pattern. All the tumors expressed CD117. One patient presented with local recurrence and metastasis, one was lost to the follow-up and two patients were doing well. Immunohistochemistry is useful in distinguishing EGIST from other mesenchymal tumors. Intra-abdominal aggressive fibromatosis may express actin and CD117 but catenin is also positive, leiomyosarcoma expresses the actin and/or desmin but CD117 is usually negative, retroperitoneal dedifferentiated liposarcoma is CD117 negative and PS100 positive, inflammatory myofibroblastic tumor is negative for CD117 and CD34. The solitary fibrous tumor expresses CD34 and is negative to CD117. Some tumors expressing CD117 such desmoplastic round cell tumor should not be confused with EGIST. On the occasion of these four observations, we will discuss the clinical aspects and the main differential diagnoses of this tumor. PMID- 16472760 TI - Herbicide binding and thermal stability of photosystem II isolated from Thermosynechococcus elongatus. AB - Binding of herbicides to photosystem II inhibits the electron transfer from Q(A) to Q(B) due to competition of herbicides with plastoquinone bound at the Q(B) site. We investigated herbicide binding to monomeric and dimeric photosystem II core complexes (PSIIcc) isolated from Thermosynechococcus elongatus by a combination of different methods (isothermal titration and differential scanning calorimetry, CD spectroscopy and measurements of the oxygen evolution) yielding binding constants, enthalpies and stoichiometries for various herbicides as well as information regarding stabilization/destabilization of the complex. Herbicide binding to detergent-solubilized PSIIcc can be described by a model of single independent binding sites present on this important membrane protein. Interestingly, binding stoichiometries herbicide:PSIIcc are lower than 1:1 and vary depending on the herbicide under study. Strong binding herbicides such as terbutryn stabilize PSIIcc in thermal unfolding experiments and endothermically binding herbicides like ioxynil probably cause large structural changes accompanied with the binding process as shown by differential scanning calorimetry experiments of the unfolding reaction of PSIIcc monomer in the presence of ioxynil. In addition we studied the occupancy of the Q(B) sites with plastoquinone (PQ9) by measuring flash induced fluorescence relaxation yielding a possible explanation for the deviations of herbicide binding from a 1:1 herbicide/binding site model. PMID- 16472761 TI - EGFR signaling pathway negatively regulates PSA expression and secretion via the PI3K-Akt pathway in LNCaP prostate cancer cells. AB - Epidermal growth factor (EGF) and its receptor (EGFR) are involved in hormone refractory growth and poor prognosis of a subgroup of human prostate cancer. In this communication, we investigated the regulation of PSA by the EGFR signaling pathway using LNCaP C-81 prostate cancer cells. Administration of EGF stimulated the growth of LNCaP C-81 cells, however, PSA expression and secretion were suppressed. An EGFR inhibitor, AG1478, abrogated the PSA suppression effect by EGF, in concurrence with the suppression of tyro-phosphorylation levels of EGFR. Interestingly, the AR level was also decreased in EGF-treated LNCaP C-81 cells. Moreover, LY294002, but not PD98059, inhibited the PSA and AR suppression effect by EGF in concurrence with the suppression of phosphorylation levels of Akt. In conclusion, our results strongly suggest the existence of a novel androgen independent PSA regulatory mechanism, i.e., the EGFR signaling pathway negatively regulates PSA expression which may be induced by the alteration of AR expression via the PI3K-Akt pathway in LNCaP C-81 cells. PMID- 16472762 TI - CYP1A1 in polycyclic aromatic hydrocarbon-induced B lymphocyte growth suppression. AB - The AhR is a ligand-activated transcription factor that mediates immunosuppression by environmental PAH. Previous studies demonstrated that activation of mature human B cells up-regulates AhR expression, suggesting that human B cells are direct PAH targets. To test this hypothesis and to determine the metabolic requirements for PAH toxicity in a human model, the effects of a prototypic PAH, B[a]P, on B cell growth were evaluated. B[a]P and its proximal (B[a]P-7,8-dihydrodiol) and terminal (B[a]P-7,8-dihydrodiol-9,10-epoxide) metabolites inhibited growth in a dose-dependent manner. A poorly metabolized AhR ligand had no effect, suggesting that biotransformation is required for growth inhibition. Inhibition of the CYP1A1 monooxygenase completely blocked growth inhibition induced by B[a]P or B[a]P-7,8-dihydrodiol, but not by B[a]P dihydrodiol-9,10-epoxide, indicating that CYP1A1-dependent metabolism of B[a]P into the terminal B[a]P-7,8-dihydrodiol-9,10-epoxide metabolite is required for growth inhibition. These studies show for the first time the metabolic requirements for PAH-mediated suppression of human B cell growth. PMID- 16472763 TI - Alzheimer's amyloid beta-peptide (1-42) induces cell death in human neuroblastoma via bax/bcl-2 ratio increase: an intriguing role for methionine 35. AB - The beta amyloid (Abeta), the major protein component of brain senile plaques in Alzheimer's disease, is known to be directly responsible for the production of free radicals toxic to brain tissue and the redox state of Met-35 residue seems to play a particular and critical role in peptide's neurotoxic actions. In this study, we investigated, in human neuroblastoma cells (IMR-32), the relationship between the oxidative state of methionine, and both neurotoxic and pro-apoptotic actions induced by Abeta-peptide, comparing the effects of native peptide, in which the Met-35 is present in the reduced state, with those of a modified peptide with oxidized Met-35 (Abeta(1-42)(35Met-ox)), as well as an Abeta derivative with Met-35 substituted with norleucine (Abeta(1-42)(35Nle)). The obtained results show that Abeta induces a time-dependent decrease in cell viability; Abeta(1-42)(35Met-ox) was significantly less potent, though inducing a remarkable decrease in cell viability compared to control. On the contrary, no toxic effects were observed after treatment with Abeta(1-42)(35Nle). Abeta peptide as well as the amyloid modified peptide with oxidized Met-35 induced the pro-apoptotic gene bax over-expression after 24 h, whereas Abeta(1-42)(35Nle) had no effect. Conversely, bcl-2, an anti-apoptotic gene, became highly down regulated by Abeta peptide treatment, in contrast to that evidenced by the Abeta(1-42)(35Met-ox) peptide. Finally, Abeta caused an increase in caspase-3 activity to be higher with respect to that shown by Abeta(1-42)(35Met-ox) while Abeta(1-42)(35Nle) had no effect. These results support the hypothesis that Abeta induced neurotoxicity occurs via bax over-expression, bcl-2 down-regulation, and caspase-3 activation, first indicating that methionine 35 redox state may alter this cell death pathway. PMID- 16472764 TI - rmlB and rmlC genes are essential for growth of mycobacteria. AB - The rhamnose-GlcNAc disaccharide linker is fundamental to the structural integrity of mycobacterial cell wall. The donor dTDP-rhamnose is synthesized by four enzymes (RmlA, B, C, and D) beginning with dTTP and glucose-1-phosphate. We generated M. smegmatis rmlB gene knockout mutant (transcription of downstream rmlC gene was blocked because of a polar effect) by homologous recombination. When the Mycobacterium tuberculosis (Tb) rmlB rescue plasmid carrying a temperature-sensitive replication origin and Tb rmlC bearing plasmid with a normal replication origin were present in the mc(2)155 rmlB knockout mutant, the mutant was unable to grow at non-permissive temperature (42 degrees C) where the Tb rmlB rescue plasmid is lost. While the Tb rmlC rescue plasmid carrying a temperature-sensitive replication origin and Tb rmlB bearing plasmid with a normal replication origin were present in the mc(2)155 rmlB knock out mutant, this mutant was also unable to grow at the non-permissive temperature where the Tb rmlC rescue plasmid is lost. These results demonstrate that rmlB and rmlC genes are essential for mycobacterial growth, therefore, RmlB and RmlC are essential targets to develop new anti-tuberculosis drugs. PMID- 16472765 TI - Structural and functional characterization of peptide-beta2m fused HLA A2/MART1(27-35) complexes. AB - The uses of soluble HLA class I/peptide complexes to monitor antigen reactive T cells are often hampered by their low-yield and high-cost production. As an alternative strategy, the peptide-beta(2)m fused, 2-component (2C) HLA class I/peptide complex has been developed, but its application is limited due to the lack of the comparison of its structural and functional characteristics with those of its conventional 3-component (3C) counterpart. In this study, we have demonstrated that the 2C and 3C HLA-A2/MART1(27-35) complexes have a similar chromatographical profile and comparable stability, but the former has 2.5 times higher yield and significantly higher binding ability with HLA-A2/MART1(27-35) complex-specific receptors than the latter. Furthermore, the 2C complex has a comparable ability to stimulate specific CTL proliferation, but appears to be more effective in eliciting the cytotoxicity of antigen-specific CTL, as compared to its 3C counterpart. PMID- 16472766 TI - UnpEL/Usp4 is ubiquitinated by Ro52 and deubiquitinated by itself. AB - The autoantigen Ro52 is an E3 ubiquitin ligase that can ubiquitinate itself (self ubiquitination). Recently, we showed that UnpEL/Usp4 is an isopeptidase that can deconjugate ubiquitin from self-ubiquitinated Ro52. Here, we showed that UnpEL is ubiquitinated by Ro52 in cooperation with UbcH5B in vitro. We also showed that UnpEL is ubiquitinated by Ro52 in HEK293T cells. Interestingly, a catalytically inactive UnpEL mutant was strongly ubiquitinated by Ro52 in HEK293T cells. These results suggest that wild-type UnpEL is ubiquitinated by Ro52 and deubiquitinated by itself (self-deubiquitination), while mutant UnpEL is ubiquitinated by Ro52 but not deubiquitinated by itself. In conclusion, Ro52 and UnpEL transregulate each other by ubiquitination and deubiquitination. PMID- 16472768 TI - Cytochrome P450 enzymes from the metabolically diverse bacterium Rhodopseudomonas palustris. AB - Four (CYP195A2, CYP199A2, CYP203A1, and CYP153A5) of the seven P450 enzymes, and palustrisredoxin A, a ferredoxin associated with CYP199A2, from the metabolically diverse bacterium Rhodopseudomonas palustris have been expressed and purified. A range of substituted benzenes, phenols, benzaldehydes, and benzoic acids was shown to bind to the four P450 enzymes. Monooxygenase activity of CYP199A2 was reconstituted with palustrisredoxin A and putidaredoxin reductase of the P450cam system from Pseudomonas putida. We found that 4-ethylbenzoate and 4 methoxybenzoate were oxidized to single products, and 4-methoxybenzoate was demethylated to form 4-hydroxybenzoate. Crystals of substrate-free CYP199A2 which diffracted to approximately 2.0A have been obtained. PMID- 16472767 TI - Expression of a functional extracellular calcium-sensing receptor in human aortic endothelial cells. AB - Extracellular Ca(2+) concentration ([Ca(2+)](o)) regulates the functions of many cell types through a G protein-coupled [Ca(2+)](o)-sensing receptor (CaR). Whether the receptor is functionally expressed in vascular endothelial cells is largely unknown. In cultured human aortic endothelial cells (HAEC), RT-PCR yielded the expected 555-bp product corresponding to the CaR, and CaR protein was demonstrated by fluorescence immunostaining and Western blot. RT-PCR also demonstrated the expression in HAEC of alternatively spliced variants of the CaR lacking exon 5. Although stimulation of fura 2-loaded HAEC by several CaR agonists (high [Ca(2+)](o), neomycin, and gadolinium) failed to increase intracellular Ca(2+) concentration ([Ca(2+)](i)), the CaR agonist spermine stimulated an increase in [Ca(2+)](i) that was diminished in buffer without Ca(2+) and was abolished after depletion of an intracellular Ca(2+) pool with thapsigargin or after blocking IP(3)- and ryanodine receptor-mediated Ca(2+) release with xestospongin C and with high concentration ryanodine, respectively. Spermine stimulated an increase in DAF-FM fluorescence in HAEC, consistent with NO production. Both the increase in [Ca(2+)](i) and in NO production were reduced or absent in HAEC transfected with siRNA specifically targeted to the CaR. HAEC express a functional CaR that responds to the endogenous polyamine spermine with an increase in [Ca(2+)](i), primarily due to release of IP(3)- and ryanodine sensitive intracellular Ca(2+) stores, leading to the production of NO. Expression of alternatively spliced variants of the CaR may result in the absence of a functional response to other known CaR agonists in HAEC. PMID- 16472769 TI - Two GC-rich boxes in huC promoter play distinct roles in controlling its neuronal specific expression in zebrafish embryos. AB - HuC, a vertebrate ortholog of Drosophila elav gene, encodes an RNA binding protein and is involved in early neurogenesis. Zebrafish huC is expressed in distinct neurons, including Rohon-Beard (RB) sensory neurons, interneurons and motoneurons, during primary neurogenesis, and in all neurons later during secondary neurogenesis. In this study, we identify two GC-rich box elements, proximal GC (p-GC) box from -172 to -149 and distal GC (d-GC) box from -218 to 208, in zebrafish huC promoter. Using transgenic approach, we demonstrate that deletion of the p-GC box from the promoter results in loss of expression of the reporter GFP in neurons while deletion of the d-GC box leads to GFP expression only in dorsal RB sensory neurons. These results suggest that the p-GC box alone confers transcriptional activity of huC promoter in primary RB neurons and the d GC is required for huC transcription in the full spectrum of spinal cord neurons. Further studies are needed to identify specific Sp1-like transcription factors that bind to these GC boxes and activate huC transcription. PMID- 16472770 TI - Identification of differentially expressed genes in human heart with ventricular septal defect using suppression subtractive hybridization. AB - Ventricular septal defect (VSD) accounts for the largest number of birth congenital heart defects in human, but the genetic programs that control ventricular septation are poorly understood. To identify differentially expressed genes between ventricular septal defect and normal ventricular septum myocardium, we have undertaken suppression subtractive hybridization (SSH) and generated reciprocal cDNA collections of representative mRNAs specific to human heart with ventricular septal defect versus normal control. Following SSH, 1378 clones were sequenced and found to derive from 551 different genes. These predominately expressed genes included genes involved in energy metabolism, cell cycle and growth, cytoskeleton and cell adhesion, LIM protein, zinc finger protein, and development. It is anticipated that further study of genes identified will provide insights into their specific roles in the etiology of VSD, even in cardiac development, aging, and disease. PMID- 16472771 TI - Structural separation of different extracellular activities in aminoacyl-tRNA synthetase-interacting multi-functional protein, p43/AIMP1. AB - AIMP1 (previously known as p43) is first found as a factor associated with a macromolecular tRNA synthetase complex. However, it is also secreted and acts on diverse target cells such as endothelial cells, macrophages, and fibroblasts to control angiogenesis, inflammation, and dermal regeneration, respectively. We previously showed that AIMP1 induces the death of endothelial cell but proliferation of fibroblasts and activates macrophages. In this work, we found that elastase 2-cleaved AIMP1 retained its pro-apoptotic activity to endothelial cells but lost the growth-stimulatory activity to fibroblasts. To determine the functional domains responsible for each activity, we generated several deletion fragments of AIMP1 and compared the activities to the target cells. AIMP1 promoted endothelial cell death and caspase-3 activation through its 101-114 amino acid region, fibroblast proliferation through its 6-46 amino acid region, and endothelial migration through its 114-192 amino acid region as revealed by deletion mapping. Thus, this work revealed that AIMP1 uses different regions for its diverse extracellular activities. PMID- 16472772 TI - Regulation of IRF7 through cell type-specific protein stability. AB - Interferon regulatory factor 7 (IRF7) is a key component of the cellular response to virus infection that culminates in physiologically relevant IFNalpha production. We studied molecular mechanisms governing responses to respiratory viral infection that are characterized by transient induction and subsequent shut off of interferon (IFN) gene expression. We asked whether alterations in IRF7 protein stability occurred during virus infection that might contribute to this regulation. To this end, we measured IRF7 half-life in various cell types and found it to be short-lived, in marked contrast to the pronounced stability of the related transcription factor, IRF3. Furthermore, virus infection accelerated IRF7 degradation in a proteosome-dependent manner in most cell types. However, plasmacytoid dendritic cells (pDC), which constitute the major circulating IFN producing cell type, displayed a distinct pattern of regulation. Infection of lymphoid tissues, where the majority of IRF7 is expressed in pDC, attenuated the normal proteosome-mediated degradation of IRF7, resulting in a long-lived protein. Stabilization was partially stimulated by autocrine IFN as a positive feedback mechanism, but was partially IFN independent. Thus, two distinct posttranslational mechanisms regulate IRF7 activity in response to viral infection, with protein turnover attenuating responses postinfection in most cell types while infection-induced protein stabilization contributes to the heightened IFN production characteristic of pDC. PMID- 16472774 TI - Oxidative stress promotes mutant huntingtin aggregation and mutant huntingtin dependent cell death by mimicking proteasomal malfunction. AB - Huntington's disease (HD) is a familial neurodegenerative disorder caused by an abnormal expansion of CAG repeats in the coding region of huntingtin gene. A major hallmark of HD is the proteolytic production of N-terminal fragments of huntingtin containing polyglutamine repeats that form ubiquitinated aggregates in the nucleus and cytoplasm of the affected neurons. However, the mechanism by which the mutant huntingtin causes neurodegeneration is not well understood. Here, we found that oxidative stimuli enhance the polyglutamine-expanded truncated N-terminal huntingtin (mutant huntingtin) aggregation and mutant huntingtin-induced cell death. Oxidative stimuli also lead to rapid proteasomal dysfunction in the mutant huntingtin expressing cells as compared to normal glutamine repeat expressing cells. Overexpression of Cu/Zn superoxide dismutase (SOD1), Hsp40 or Hsp70 reverses the oxidative stress-induced proteasomal malfunction, mutant huntingtin aggregation, and death of the mutant huntingtin expressing cells. Finally, we show the higher levels of expression of SOD1 and DJ 1 in the mutant huntingtin expressing cells. Our result suggests that oxidative stress-induced proteasomal malfunction might be linked with mutant huntingtin induced cell death. PMID- 16472773 TI - Responses of brain and non-brain endothelial cells to meningitis-causing Escherichia coli K1. AB - Bacterial interaction with specific host tissue may contribute to its propensity to cause an infection in a particular site. In this study, we examined whether meningitis-causing Escherichia coli K1 interaction with human brain microvascular endothelial cells, which constitute the blood-brain barrier, differed from its interaction with non-brain endothelial cells derived from skin and umbilical cord. We showed that E. coli K1 association was significantly greater with human brain microvascular endothelial cells than with non-brain endothelial cells. In addition, human brain microvascular endothelial cells maintained their morphology and intercellular junctional resistance in response to E. coli K1. In contrast, non-brain endothelial cells exhibited decreased transendothelial electrical resistance and detachment from the matrix upon exposure to E. coli K1. These different responses of brain and non-brain endothelial cells to E. coli K1 may form the basis of E. coli K1's propensity to cause meningitis. PMID- 16472775 TI - Single amino-acid InDel variants generated by alternative tandem splice-donor and -acceptor selection. AB - We have investigated putative single amino-acid InDel variants with human ESTs. Examination of the formation process for single amino-acid InDel variants indicates a possible splicing mechanism in addition to the genomic insertion/deletion events as would be expected. The wobble-splicing transcripts were often generated around the intron-exon boundaries by selecting an alternative neighboring splice signal sequence, in particular the tandem agNAG or GTNgt sequence at the splice-acceptor or -donor site, thus creating single amino acid InDel isoforms. Another category of variants was identified with one altered amino-acid plus one amino-acid InDel, under divergent coding-frame usage. We demonstrate that such minute distance of splice site choice generates an even greater level of transcriptome diversity, and suggest that non-functional synonymous or intronic SNPs could be converted to functionally significant InDel alterations through this process. This subtle alteration in mRNA and protein coding sequence may elicit a great impact upon human genome and proteome diversity. PMID- 16472776 TI - Identification of integrin alpha1 as an interacting protein of protein tyrosine phosphatase PRL-3. AB - PRL-3 is a newly identified protein tyrosine phosphatase associated with tumor metastasis. It is over-expressed in various cancers, such as colorectal cancer, gastric cancer, and ovarian cancer, and is correlated with the progression and survival of cancers. Although PRL-3 plays a causative role in promoting cancer cell invasion and metastasis, the molecular mechanism is unknown. To investigate PRL-3's roles in tumorigenesis and signal transduction pathway, we screened the human placenta brain cDNA library with the bait of PRL-3 in yeast two-hybrid system. Then we identified integrin alpha1 as a PRL-3-interacting protein for the first time, and verified this physical association with pull-down and co immunoprecipitation assays. Furthermore, we found that PRL-3 could down-regulate the tyrosine-phosphorylation level of integrin beta1 and increased the phosphorylation level of Erk1/2. Our present discovery will provide new clues for elucidating the molecular mechanism of PRL-3 in promoting cancer invasion and metastasis. PMID- 16472777 TI - Novel short chain fatty acids restore chloride secretion in cystic fibrosis. AB - Phenylalanine deletion at position 508 of the cystic fibrosis transmembrane conductance regulator (DeltaF508-CFTR), the most common mutation in cystic fibrosis (CF), causes a misfolded protein exhibiting partial chloride conductance and impaired trafficking to the plasma membrane. 4-Phenylbutyrate corrects defective DeltaF508-CFTR trafficking in vitro, but is not clinically efficacious. From a panel of short chain fatty acid derivatives, we showed that 2,2-dimethyl butyrate (ST20) and alpha-methylhydrocinnamic acid (ST7), exhibiting high oral bioavailability and sustained plasma levels, correct the DeltaF508-CFTR defect. Pre-incubation (>or=6h) of CF IB3-1 airway cells with >or=1mM ST7 or ST20 restored the ability of 100microM forskolin to stimulate an (125)I(-) efflux. This efflux was fully inhibited by NPPB, DPC, or glibenclamide, suggesting mediation through CFTR. Partial inhibition by DIDS suggests possible contribution from an additional Cl(-) channel regulated by CFTR. Thus, ST7 and ST20 offer treatment potential for CF caused by the DeltaF508 mutation. PMID- 16472778 TI - Thymidine secretion by hybridoma and myeloma cells. AB - Secretion of thymidine appeared to be a common property of hybridoma and myeloma cells, but not of other cell types, which were tested. Of three hybridoma cell lines tested, all secreted thymidine in amounts resulting in the accumulation of thymidine to concentrations of 10-20microM in the culture medium. Also three of five myeloma cell lines that were analyzed secrete thymidine, but none of the other cell types that were studied. Thymidine was purified to homogeneity (4mg purified from 3l of culture medium) and identified as such by nuclear magnetic resonance spectroscopy. The cells that secreted thymidine showed high resistance to the growth inhibitory effect of thymidine. PMID- 16472779 TI - Arabidopsis cytosolic glutamine synthetase AtGLN1;1 is a potential substrate of AtCRK3 involved in leaf senescence. AB - While considerable progress has been achieved in plant CDPK studies in the past decade, there is relatively no information about the potential substrates of CRKs. In this report, a yeast two-hybrid screen was performed with truncated form of AtCRK3 as bait to identify its interacting proteins in an effort to dissect its physiological roles. One gene encoding cytosolic glutamine synthetase AtGLN1;1 was isolated. Further analyses indicated that AtGLN1;1 could interact specifically with AtCRK3 and the kinase domain of AtCRK3 and the catalytic domain of AtGLN1;1 were responsible for such interaction, respectively. Furthermore, in vitro and in vivo co-immunoprecipitation results strongly supported that they could physically interact with each other. Phosphorylation assays revealed that AtGLN1;1 could be specifically phosphorylated by AtCRK3 in vitro. All the results demonstrate that AtGLN1;1 may be a substrate of AtCRK3. In addition, both AtGLN1;1 and AtCRK3 could be induced by natural or artificially induced leaf senescence, implying that such interaction may be involved in the regulation of nitrogen remobilization during leaf senescence. PMID- 16472780 TI - 3'-Azido-3'-deoxythymidine (AZT) inhibits thymidine phosphorylation in isolated rat liver mitochondria: a possible mechanism of AZT hepatotoxicity. AB - 3'-azido-3'-deoxythymidine (AZT) is a staple of highly active antiretroviral therapy (HAART). Prior to HAART, long-term use of high-dosage AZT caused myopathy, cardiomyopathy, and hepatotoxicity, associated with mitochondrial DNA depletion. As a component of HARRT, AZT causes cytopenias and lipodystrophy. AZT 5'-triphosphate (AZTTP) is a known inhibitor of the mitochondrial polymerase gamma and has been targeted as the source of the mitochondrial DNA depletion. However, in previous work from this laboratory with isolated rat heart mitochondria, AZT phosphorylation beyond AZT-5'-monophosphate (AZTMP) was not detected. Rather, AZT was shown to be a more potent inhibitor of thymidine phosphorylation (50% inhibitory concentration (IC50) of 7.0+/-1.0 microM) than AZTTP is of polymerase gamma (IC50 of >100 microM), suggesting that depletion of mitochondrial stores of TTP may limit replication. This work has investigated whether an identical mechanism might account for the hepatotoxicity seen with long-term use of AZT. Isolated rat liver mitochondria were incubated with labeled thymidine or AZT, and the rate and extent of phosphorylation were determined by HPLC analysis of acid-soluble extracts of the incubated mitochondria. The results showed that in the phosphorylation of thymidine to TMP, liver mitochondria exhibit a higher Vmax and Km than heart mitochondria, but otherwise heart and liver mitochondria display similar kinetics. AZT is phosphorylated to AZTMP, but no further phosphorylated forms were detected. In addition, AZT inhibited the production of TTP, with an IC50 of 14.4+/-2.6 microM AZT. This is higher, but comparable to, the results seen in isolated rat heart mitochondria. PMID- 16472781 TI - Insulin sensitizers may prevent metabolic inflammation. AB - The relative decreased response of peripheral tissues to insulin (insulin resistance) is a key metabolic disturbance that predisposes a large percentage of individuals to the development of type 2 diabetes and to cardiovascular disease. As detailed in an extensive literature over the last two decades, insulin resistance co-exists in varying degrees with a variety of other key risk factors, including dyslipidemia, hypertension, and vascular inflammation, that contribute to poor cardiovascular outcomes of individuals with type 2 diabetes and metabolic syndrome. Whereas insulin resistance is generally thought of as pathology unto itself, this commentary suggests that insulin resistance is a physiological compensation to inappropriate oxidative metabolism that induces a metabolic inflammatory response. Via signaling of this inflammatory response, the protective compensation to excessive oxidative metabolism dampens metabolism by reducing insulin action, fatty acid oxidation, and eventually mitochondrial function and numbers. Such a scenario could explain the coexistence of these phenomena with obesity and reduced mitochondrial function. Recent evidence suggests that thiazolidinediones exert pharmacology through modifications of mitochondrial metabolism, preventing the metabolic inflammation and allowing the up regulation of mitochondrial biogenesis. A further understanding of these mechanisms, which are likely to involve key redox signaling events emanating from mitochondrial biochemistry, is needed to fuel new therapeutic advances for the treatment of metabolic syndrome. PMID- 16472782 TI - Estrogen and CCK1 receptor modification of mu-opioid receptor binding in the cortex of female rats. AB - Cholecystokinin (CCK) in the nervous system has effects opposite to those of opioids. However, the mechanism by which CCK opposes the effect of opioids at the receptor or cellular level is still unknown. In the brain, distributions of CCK receptors and opioid receptors have been demonstrated to overlap. The present study was undertaken to determine the mechanism of CCK-opioid interactions in the cortex of ovariectomized rats. Furthermore, because estrogen is a powerful regulator of CCK and opioid activity, we examined whether estrogen state also modulates the interactions of these neuropeptides. mu-Opioid (MOP) receptor binding was examined in cortical membranes that were preincubated with CCK-8S and CCK receptor agonist and antagonist followed with 3H-DAMGO. Pharmacological results revealed that CCK-8S suppressed 3H-DAMGO binding in cortical membranes of ovariectomized rats. The same result was obtained using a CCK1 receptor agonist (JMV-180), whereas a CCK2 receptor agonist (CCK-4) failed to suppress 3H-DAMGO binding. Antagonism of the CCK1 receptor by JMV-179 blocked both CCK-8S and JMV 180 suppression of 3H-DAMGO binding. Furthermore, estrogen treatment to female rats resulted in a suppression of 3H-DAMGO binding in cortical membranes. These results demonstrate an estrogen regulation of the MOP receptor and a protein protein interaction between CCK1 receptor and MOP receptor. PMID- 16472783 TI - In humans Ib facilitation depends on locomotion while suppression of Ib inhibition requires loading. AB - The role of force feedback during gait is still a matter of debate. From work on cats, it is known that input from Golgi tendon organs from triceps surae does produce Ib facilitation during locomotion instead of autogenic inhibition. In humans, Stephens and Yang (Stephens, M.J., Yang, J.F., 1996. Short latency, non reciprocal group I inhibition is reduced during the stance phase of walking in humans. Brain Res. 743, 24-31) found that voluntary contraction results in a reduction of Ib inhibition. During gait, they even observed Ib facilitation in a subset of subjects. This raises the question whether the crucial elements involved in these changes are either loading of the leg or locomotion. To examine this question, Ib reflexes were investigated during sitting, lying supine, lying supine with 300 N pressure applied to the foot sole, standing, and a rhythmic loading and unloading task called "reduced" gait. Ib inhibition was obtained during sitting and lying supine. This inhibition was significantly reduced or disappeared during standing and when lying supine but loaded. During the stance phase of "reduced" gait, the inhibition disappeared in eight subjects, and even a facilitation was observed in six subjects. It is concluded that the decrease in Ib inhibition from gastrocnemius to soleus occurs during a load-bearing condition and does not require locomotion. In contrast, Ib facilitation requires locomotion at least in a rudimentary form. PMID- 16472784 TI - Effects of prolonged hypobaric hypoxia on carotid nerve endings and glomus cells. Changes in intercellular coupling. AB - Carotid bodies were removed from anesthetized rats kept under normobaric (640 Torr) and hypobaric conditions (380 Torr for 2-3 weeks). Slices (100-150 microm) of the organ were viewed under an inverted microscope for simultaneous stimulation and recording of coupled glomus cells and carotid nerve endings. The latter were identified by their more negative Em, high input resistance (Ro) and time-dependent rectification in response to negative current pulses. Also, when nerve endings had an Em more negative than -40 mV showed spontaneous activity in the form of mini-receptor potentials (mrps). Glomus cells had less negative Em and lower Ro. Prolonged hypobaric hypoxia did not change the Em of nerve endings and glomus cells. However, in both structures, Ro increased. Also, the mrps became smaller and occurred less frequently. Intercellular coupling was recognized when currents applied to one cell spread to adjoining ones. In the case of glomus cells (GC/GC coupling), it was mostly resistive and bidirectional. Coupling between nerve endings and glomus cells was more complex, When a glomus cell was stimulated, current spread to the nerve ending (GC/NE coupling) was similar in magnitude (2-3%) to coupling between GCs. However, when NE was stimulated current spread to GC (NE/GC coupling) was minimal (less than 0.1%) and transient (capacitive). Nerve endings were also bidirectionally and capacitively coupled (NE/NE coupling) with a median of 2,8%. Intracellularly injected Lucifer Yellow or Alexa 488 diffused to neighboring structures. Prolonged hypobaric hypoxia significantly tightened coupling modes GC/NE, NE/GC, and NE/NE but reduced GC/GC coupling. Tighter coupling was accompanied by lower coupling resistance, and the opposite occurred when intercellular coupling decreased. Increased GC/NE and reduced GC/GC coupling during hypobaric hypoxia may be partly responsible for the increased reactivity of these receptors under this condition. PMID- 16472785 TI - NMDA-induced preconditioning attenuates synaptic plasticity in the rat hippocampus. AB - It was recently demonstrated that glutamate could precondition hippocampal slices against the damaging effects of hypoxia, and we have now extended this observation by investigating (i) the ability of glutamate receptor agonists to act as preconditioning agents and (ii) the effects of preconditioning on synaptic plasticity. Using rat hippocampal slices, 15 microM NMDA applied for 10 min (chemical insult) caused abolition of the population spike potentials (PS) followed by approximately 33% recovery at 60 min post-insult. In comparison, a 5 min preconditioning exposure of 10 microM NMDA given 30 min prior to the insult significantly improved the recovery to 69%. Preconditioning did not alter paired pulse facilitation; however, it significantly enhanced paired pulse depression and reduced population spike long-term potentiation (PS-LTP) and LTP in field recordings. This effect on PS-LTP appeared to be NMDA receptor dependent and was blocked by the nitric oxide synthase inhibitors nitro-L-arginine methyl ester (L NAME) and 7-nitro indazole (7-NI) but not by the adenosine receptor antagonist 8 cyclopentyl-1,3-dipropylxanthine (DPCPX). We conclude that preconditioning by NMDA can improve recovery following acute insults but may have deleterious effects on neuronal plasticity. PMID- 16472786 TI - Cannabinoid CB2 receptors: immunohistochemical localization in rat brain. AB - Brain expression of CB2 cannabinoid receptors has been much less well established and characterized in comparison to the expression of brain CB1 receptors. Since CB2 receptors are intensely expressed in peripheral and immune tissues, expression in brain microglia has been anticipated. Nevertheless, we now describe expression of CB2-receptor-like immunoreactivity in brain in neuronal patterns that support broader CNS roles for this receptor. Two anti-CB2 affinity purified polyclonal antibodies were raised in rabbits immunized with peptide conjugates that corresponded to amino acids 1-33 and 20-33. Western blot analyses revealed specific bands that were identified using these sera and were absent when the sera were preadsorbed with 8.3 mug/ml of the immunizing peptides. These studies, and initial RT-PCR analyses of brain CB1 and CB2 mRNAs, also support the expression of brain CB2 receptor transcripts at levels much lower than those of CB1 receptors. CB2 cannabinoid receptor mRNA was clearly expressed in the cerebellum of wild type but not in CB2 knockout mice. CB2 immunostaining was detected in the interpolar part of spinal 5th nucleus of wild type but not in CB2 knockout mice, using a mouse C-terminal CB2 receptor antibody. Immunohistochemical analyses revealed abundant immunostaining for CB2 receptors in apparent neuronal and glial processes in a number of rat brain areas. Cerebellar Purkinje cells and hippocampal pyramidal cells revealed substantial immunoreactivity that was absent when sections were stained with preadsorbed sera. CB2 immunoreactivity was also observed in olfactory tubercle, islands of Calleja, cerebral cortex, striatum, thalamic nuclei, hippocampus, amygdala, substantia nigra, periaqueductal gray, paratrochlear nucleus, paralemniscal nucleus, red nucleus, pontine nuclei, inferior colliculus and the parvocellular portion of the medial vestibular nucleus. In-vitro, CB2 immunoreactivity was also present in hippocampal cell cultures. The multifocal expression of CB2 immunoreactivity in glial and neuronal patterns in a number of brain regions suggests reevaluation of the possible roles that CB2 receptors may play in the brain. PMID- 16472787 TI - Multivalent glycomimetics: synthesis of nonavalent mannoside clusters with variation of spacer properties. AB - Oligosaccharide mimetics are important tools in the glycosciences. In this work, we have employed spaced glycodendrons for the synthesis of oligomannoside mimetics. Starting from a number of trivalent, branched molecular wedges, the preparation of nonavalent cluster mannosides was accomplished, which were varied with regard to the chemical characteristics of their spacer moieties and spacer lengths. For ligation of the various trivalent dendrons to the nonavalent target molecules peptide coupling was employed. PMID- 16472789 TI - Controlled peeling of the surfaces of starch granules by gelatinization in aqueous dimethyl sulfoxide at selected temperatures. AB - Microscopic examination of starch granules in 90:10 (v/v) Me(2)SO-H(2)O indicated that the granules were slowly being gelatinized from their surfaces. The rate of gelatinization was dependent on two variables: (1) the amount of water in Me(2)SO and (2) the temperature. An increase of water in Me(2)SO and/or an increase in temperature increased the rate of gelatinization and vice versa. Specific ratios of Me(2)SO and H(2)O (85:15-95:5) and temperatures (0-15 degrees C) were found to give controlled sequential peeling/gelatinization of eight kinds of starch granules in 1-12h, with amounts of 10-25% gelatinization per hour. It was observed that the percent of starch granule remaining versus time gave curves that were linear and others that had linear parts separated by one or more abrupt changes. No two starches had a similar gelatinization curve for the same two conditions of the amount of water and the temperature. It is hypothesized that these curves reflect different structural characteristics for the individual kinds of starch granules. PMID- 16472788 TI - A highly concise preparation of O-deacetylated arylthioglycosides of N-acetyl-D glucosamine from 2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-alpha-D-glucopyranosyl chloride and aryl thiols or disulfides. AB - An expedient and mild route to a range of aryl 2-acetamido-2-deoxy-1-thio-beta-D glucopyranosides has been devised from 2-acetamido-3,4,6-tri-O-acetyl-2-deoxy alpha-D-glucopyranosyl chloride and arylthiols or aryl disulfides using phase transfer catalysis conditions. This simple procedure compresses up to three synthetic steps into a one-pot reaction, obviating the need for tedious workups and chromatography and directly furnishes crystalline materials in good yields. The procedure is compatible with a range of thiols and disulfides and may be amenable for preparing a wide range of thioglycosides with various glycons and aglycons. PMID- 16472790 TI - The structure of the O-polysaccharide from the lipopolysaccharide of Providencia alcalifaciens O30. AB - The O-polysaccharide was obtained by mild acid degradation of the lipopolysaccharide of Providencia alcalifaciens O30. Studies by sugar and methylation analyses along with (1)H and (13)C NMR spectroscopy, including two dimensional (1)H,(1)H COSY, TOCSY, ROESY, and H-detected (1)H,(13)C HSQC, HMBC, and HMQC-TOCSY experiments, showed that the polysaccharide has a linear pentasaccharide repeating unit of the following structure: PMID- 16472791 TI - Targeting bone marrow to treat vascular diseases: Accelerated vascular healing by colony stimulating factor. PMID- 16472792 TI - Potentiation of humoral immune response and activation of NF-kappaB pathway in lymphocytes in experimentally induced hyperthyroid rats. AB - This study explored the effect of hyperthyroid state on humoral immune response and NF-kappaB signaling in lymphocytes. Male Wistar rats were treated with l thyroxin for four weeks. Animals were immunized with sheep red blood cells (SRBC) after three weeks of l-thyroxin treatment. After one week of immunization, serum anti-SRBC titer was measured and NF-kappaB signaling was studied in lymphocytes by Western blot analysis of p-IKB-alpha, IKB-alpha, and p65. These results were compared with that of control rats. Antibody response and density of p-IKB-alpha and p65 were significantly higher in l-thyroxin treated rats in comparison to controls. The antibody response was found to have significant correlation with density of p-IKB-alpha and p65. Our results indicate that NF-kappaB signaling pathway in lymphocytes is activated in hyperthyroid state which might play a role in potentiation of antibody response. PMID- 16472793 TI - Specific antihepatocellular carcinoma T cells generated by dendritic cells pulsed with hepatocellular carcinoma cell line HepG2 total RNA. AB - Dendritic cell (DC) vaccination with the use of total tumor RNA provides the potential to generate a polyclonal immune response to multiple known and unknown tumor antigens without HLA restriction. Our study evaluated this approach as potential immunotherapy for patients with hepatocellular carcinoma (HCC). Immature DCs generated from peripheral blood mononuclear cells of patients with HCC were transfected with HepG2-GFP (HepG2 cells transfected stably with plasmid pEGFP-C3) cells total RNA. Transfected, matured DCs were used to stimulate autologous T cells. Results revealed that DCs transfected with HepG2-GFP cells total RNA expressed EGFP when observed by flow cytometry. Compared with those before transfection, the expressions of membrane molecules were increased dramatically, and interleukin-12p70 release in the supernatant was elevated significantly. Specific T cells generated by DCs transfected with HepG2-GFP total RNA recognized HLA-matched HepG2 cell lines specifically. These findings indicate that these RNA-transfected DCs successfully generate specific T cells that specifically recognize HCC cells. Total tumor RNA-pulsed DCs may have potential as an adjuvant immunotherapy for patients with HCC. PMID- 16472794 TI - Naproxen-induced oxidative stress in the isolated perfused rat liver. AB - We previously showed that naproxen induced the oxidative stress in the liver microsomes and the isolated hepatocytes of rats. In this study, the in situ effect of naproxen on the rat liver tissue was investigated, using the isolated perfused liver from the view-point of the naproxen-induced hepatotoxicity. The leakage of glutamic-oxaloacetic transaminase (GOT) from the perfused liver and appearance of thiobarbituric acid reactive substances (TBARS) in the perfusate increased with the progress of perfusion after a lag time of about 1h. The naproxen-perfusion of the liver decreased the biliary excretion of glutathione (GSH) and oxidized glutathione, glutathione disulfide (GSSG) prior to TBARS production and GOT leakage. GSSG content in the naproxen-perfused liver was significantly higher than in the control. TBARS appeared in the perfusate of the naproxen-perfused liver for 30 min, but not in the control. The biliary excretion clearance (CL(bile)) of indocyanine green (ICG), a reagent for testing the liver function, in the liver perfused with naproxen decreased to a half of that in the liver perfused without naproxen. Thus, the naproxen-induced oxidative stress in the liver was shown to affect the physiological function of liver through the impairment of biliary excretion, which is recognized as a detoxification system. PMID- 16472795 TI - Influence of hormone replacement therapy on blood antioxidant enzymes in menopausal women. AB - BACKGROUND: Natural loss of estrogen occurring in menopausal process may contribute to various health problems many of them possibly related to oxidative stress. Hormone replacement therapy (HRT) is the most common treatment to attenuate menopausal disturbances. This study was aimed at evaluating the influence of HRT on the activity of antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT; and glutathione peroxidase, GPx) and lipid peroxidation (thiobarbituric acid reactive substances, TBARS) in menopausal women. METHODS: Blood antioxidant enzyme activities were determined in premenopausal (n=18) and in postmenopausal healthy women without (n=21) or with (n=19) HRT (mean ages: 47, 59, and 57 years, respectively). RESULTS: TBARS, CAT, and GPx activity were not significantly different among the groups of study. However, SOD activity was significantly lower in postmenopausal women without HRT (0.68+/-0.04 U/mg Hb) when compared both to premenopausal women (0.91+/-0.04 U/mg Hb) and to postmenopausal women with HRT (0.89+/-0.07 U/mg Hb). SOD activity was positively correlated to the duration of HRT in the postmenopausal groups (r=0.33, p<0.05). CONCLUSIONS: HRT antagonizes the decrease of SOD activity that occurs after menopause, suggesting that HRT may play a beneficial role in the protection against oxidative stress. PMID- 16472796 TI - Diagnostic accuracy of serum asialo-alpha1-acid glycoprotein concentration for the differential diagnosis of liver cirrhosis and hepatocellular carcinoma. AB - BACKGROUND: Serum asialoglycoproteins concentration are increased in patients with hepatic disease. We developed an antibody-lectin sandwich assay that is sensitive and specific to measure asialo-alpha(1)-acid glycoprotein (AsAGP) concentration in human serum and evaluated it as a biochemical marker for hepatic disease. METHODS: Serum AsAGP concentration was measured by antibody-lectin sandwich assay with 610 serum specimens of patients with hepatic disease. Serum from 41 healthy donors and 155 patients with non-hepatic disease served as negative controls. The AsAGP values were analyzed by receiver operator characteristics (ROC) curve analysis. The diagnostic accuracy of AsAGP value was compared with those of the conventional biochemical markers in the liver function test. RESULTS: Serum AsAGP concentration in 83% of patients with liver cirrhosis (LC) and 89% of patients with hepatocellular carcinoma (HCC) was increased over the cutoff value (1.33 microg/ml), indicating that an increase of serum AsAGP concentration is restricted to LC or HCC cases. The area under curve (AUC) in the ROC curve was 0.919 for LC and 0.946 for HCC. CONCLUSIONS: Serum AsAGP concentration exhibited good diagnostic accuracy as a biochemical marker for LC and HCC. The addition of AsAGP to conventional liver function tests may significantly improve the diagnosis and prognosis. PMID- 16472797 TI - The effect of different doses of fluvastatin on inflammatory markers in the early phase of acute coronary syndrome. AB - BACKGROUND: Inflammation promotes acute coronary syndromes (ACS) and ensuing clinical complications. It is well known that statins decrease the risk of coronary events and may benefit the stabilization of atherosclerotic plaque with their anti-inflammatory effects. We investigated the effects of different doses of fluvastatin on serum concentrations of high-sensitive C-reaction protein (hs CRP) and tumor necrosis factor-alpha (TNF-alpha) in the early phase of ACS. METHODS: We prospectively randomized 60 patients with ACS to 3 groups: (1) group A (n=20): were given routine therapy; (2) group B (n=20): were administrated routine therapy with 40 mg/d oral fluvastatin; (3) group C (n=20): received routine therapy with 80 mg/d oral fluvastatin. Twenty patients with stable coronary heart disease served as controls. The following-up period was 7 days. By immunoturbidimetric assay and ELISA methods the serum concentrations of hs-CRP and TNF-alpha were measured before and after therapy. RESULTS: (1) The serum concentrations of hs-CRP and TNF-alpha in patients with ACS was significantly higher than those in the control group (P<0.05). (2) After 1 week of therapy, the serum concentrations of hs-CRP and TNF-alpha were significantly lower in group B and group C (all P<0.01), especially in group C. (3) The serum concentrations of hs-CRP and TNF-alpha did not correlate to the concentrations of TC, TG, LDL-C, or HDL-C. CONCLUSION: Early fluvastatin intervention decreases dose-dependently the serum concentrations of hs-CRP and TNF-alpha of patients with ACS. The high-dose fluvastatin invention may play a stronger anti-inflammatory effect in ACS patients. The anti-inflammatory effect of fluvastatin may be beyond the lipid lowering. PMID- 16472798 TI - Analysis of GABRB2 association with schizophrenia in German population with DNA sequencing and one-label extension method for SNP genotyping. AB - OBJECTIVES: Schizophrenia (SCZ) is a complex mental disease that affects approximately 1% of the population. In this study, six SNPs in GABRB2 were genotyped for a case-control association study with the cycloid psychosis subtype of SCZ in the German population using two methods for SNP genotyping. DESIGN AND METHODS: The SNPs were genotyped by direct DNA sequencing, as well as a novel one label extension method. The results were analyzed for association with SCZ. RESULTS AND CONCLUSIONS: Significant association was found for SNPs rs1816071 and rs1816072 with SCZ susceptibility. This is consistent with our previous finding of association of SNPs in GABRB2 with SCZ susceptibility in Han Chinese. There was a total agreement between the genotyping results from one-label extensions and the results of direct DNA sequencing, thus validating the accuracy of the one label extension method of SNP genotyping. PMID- 16472799 TI - Paraoxonase 1 gene Q192R polymorphism affects stroke and myocardial infarction risk. AB - OBJECTIVES: Paraoxonase (PON1), an enzyme associated with high-density lipoprotein (HDL) particles, inhibits oxidation and atherogenesis. We sought to investigate the association of the PON1 Q192R polymorphism with stroke and heart disease. DESIGN AND METHODS: In a case control study, we genotyped 242 ischemic stroke, 231 myocardial infarction (MI), and 310 healthy control subjects, all Chinese. RESULTS: R-containing genotypes (R+) were associated with vascular disease, OR = 1.5, P = 0.03. RR was increased in MI patients who were either smokers (OR = 3.1, P = 0.01), male, or younger than 60. R+ but not RR genotypes were increased in stroke patients, particularly large artery type (OR = 2.6 and P = 0.02 for R+, OR = 1.0 for RR) or among smokers. The relative dearth of RR in stroke might be due to earlier MI or death in at-risk people, such as smokers. R+ genotypes were increased with stroke in hypertensive (OR = 2.1, P = 0.02) but not normotensive (OR = 1.0) subjects. CONCLUSIONS: PON1 192R+ genotypes were associated with stroke and MI, particularly in subsets of patients, in patterns suggesting a possible survivor effect. PMID- 16472801 TI - Altered serotonergic function of dorsal raphe nucleus in perinatally protein deprived rats: effects of fluoxetine administration. AB - We have previously described that perinatally undernourished rats showed increased locus coeruleus activity, a phenomenon reversed by repeated desipramine or fluoxetine administration. Since there is reciprocal modulation between the locus coeruleus and the dorsal raphe nucleus, and because these structures are associated with the pathophysiology of different states of anxiety, we evaluated the activity of serotonergic dorsal raphe neurons from early malnourished animals compared with controls, using in vivo extracellular single-unit recordings. The number of spontaneously active cells/track was significantly higher in protein deprived animals, although the firing rate and the sensitivity of 5-HT(1A) receptors did not differ from those of controls. Five days of fluoxetine administration (5 mg/kg/day i.p.) was able to reverse the increased number of active serotonergic cells without affecting their firing rate. Furthermore, subsensitivity of 5-HT(1A) autoreceptors developed in the same way after repeated fluoxetine administration in both control and protein-deprived animals. These results suggest that the increased noradrenergic transmission observed in protein deprived animals may induce an activation of serotonergic neurons in the dorsal raphe nucleus, and that this effect is normalized following fluoxetine treatment, which normalizes locus coeruleus activity. PMID- 16472800 TI - Role of CGRP and GABA in the hypotensive effect of intrathecally administered anandamide to anesthetized rats. AB - In urethane-anesthetized rats the intrathecal (i.t.) injection of 100 nmol anandamide produced a hypotensive effect (-19.3+/-1.6 mm Hg; n=6) that was mimicked by i.t. administration of 0.25 nmol calcitonin gene-related peptide (CGRP; -26.2+/-1.8 mm Hg, n=4). Both effects were antagonized either by the CGRP receptor antagonist CGRP(8-37) (5 nmol; i.t.) or by the gamma-aminobutyric acid (GABA)(A) receptor antagonist bicuculline (8.8 nmol, i.t) or by the GABA(B) receptor antagonist 2-hydroxy saclofen (110 nmol; i.t.). On the contrary, blockade of spinal CGRP receptors by CGRP(8-37) did not modify the hypotensive response to either the GABA(A)-receptor agonist muscimol (8.8 nmol; i.t.) or the GABA(B)-receptor agonist baclofen (100 nmol; i.t). This result suggests a unidirectional effect of CGRP on the GABAergic system. The response to anandamide remained unaltered after acute inhibition of nitric oxide (NO) synthase activity by either i.t. (1 micromol) or i.v. (10 mg/kg) injection of N(G)-nitro-L-arginine methyl ester (L-NAME), but increased significantly after long-term L-NAME administration (70 mg/kg/day; four weeks; p.o.), thus suggesting compensatory changes in cardiovascular homeostasis. It is proposed that the hypotensive effect of anandamide in urethane-anesthetized rats could involve the release of CGRP followed by the release of GABA in the spinal cord. NO does not appear to have a direct participation in the spinal mechanisms involved in the decrease of the blood pressure caused by anandamide. PMID- 16472802 TI - Stimulation of erythrocyte phosphatidylserine exposure by chlorpromazine. AB - Side effects of treatment with chlorpromazine include anaemia which could result from decreased formation or accelerated clearance of circulating erythrocytes. Recently, a novel mechanism leading to erythrocyte clearance has been disclosed. Osmotic shock, oxidative stress and glucose deprivation lead to activation of cation channels, Ca(2+) entry, activation of a Ca(2+)-sensitive erythrocyte scramblase and subsequent exposure of phosphatidylserine at the erythrocyte surface. As macrophages are equipped with phosphatidylserine receptors, they bind, engulf and degrade phosphatidylserine exposing cells. The present experiments have been performed to explore whether chlorpromazine triggers phosphatidylserine exposure of erythrocytes. The phosphatidylserine exposure was estimated from annexin binding as determined in fluorescence activated cell sort (FACS) analysis. A 24 h exposure to glucose-free medium decreased cytosolic ATP levels, decreased cellular levels of reduced glutathione (GSH) and increased annexin binding. The effect on annexin binding and ATP but not on GSH was significantly enhanced in the presence of chlorpromazine (10 microM). Higher concentrations of chlorpromazine (40 microM) increased cytosolic Ca(2+) activity. Osmotic shock and Cl(-) removal similarly increased annexin binding, effects again being enhanced in the presence of chlorpromazine. In conclusion, the present observations point to a novel side effect of chlorpromazine, i.e. increased sensitivity of erythrocytes to glucose deprivation. The effect could well contribute to the known anaemia observed in the treatment with this antipsychotic drug. PMID- 16472803 TI - Silencing of sigma-1 receptor induces cell death in human lens cells. AB - Sigma receptors have no known homology with other receptor systems, have no known natural ligands, but appear to play a critical role in a large diversity of cell functions. In the absence of a conventional pharmacology, siRNA technology provides a direct means of elucidating the major cell signaling pathways influenced by this receptor system. The non-transformed human lens cell line FHL124 was found to express the sigma-1 receptor (Sig-1R) and was employed for these studies. 72 h of transfection with either of the two siRNA directed against the sigma-1 receptor reduced messenger RNA and protein levels by over 70 and 60% respectively. Subsequent incubation for 96 h in culture medium (EMEM) supplemented with 5% serum gave a partial recovery of message, but there was no significant increase in protein. LDH leakage assays showed that significant cell death occurred during this time with an increased expression of caspase-3. Thrombin (10 nM) drives the growth of lens cells with a concomitant increase in ERK and Akt phosphorylation. These increases were inhibited in the cells where knockdown had occurred but not in cells exposed to scrambled siRNA. This study establishes a central role for Sig-1R in cell survival and death. PMID- 16472806 TI - Opisthorchis viverrini: influence of maternal infection in hamsters on offspring infected with homologous parasite and their IgG antibody response. AB - We investigated the influence in hamsters of a maternal Opisthorchis viverrini infection on their offspring infected with homologous parasites and the kinetics of the O. viverrini-specific IgG antibody responses. No significant difference (P > 0.05) was found in the specific IgG antibody response and the number of O. viverrini eggs per gram feces (EPG) between infected offspring from infected mothers and infected offspring from uninfected mothers. A significant difference (P < 0.05) of EPG per worm was found between infected offspring from infected mothers and infected offspring from uninfected mothers only when the offspring were infected with O. viverrini after weaning at 5 weeks of age. The worm loads in infected offspring from infected mothers were significantly less than that in infected offspring from uninfected mothers. This study demonstrated that maternal infection effects worm fecundity and the worm load in an infected offspring. PMID- 16472805 TI - Transplanted neural precursor cells reduce brain inflammation to attenuate chronic experimental autoimmune encephalomyelitis. AB - Stem cell transplantation was introduced as a mean of cell replacement therapy, but the mechanism by which it confers clinical improvement in experimental models of neurological diseases is not clear. Here, we transplanted neural precursor cells (NPCs) into the ventricles of mice at day 6 after induction of chronic experimental autoimmune encephalomyelitis (EAE), a model of multiple sclerosis (MS). Transplanted cells migrated into white matter tracts and attenuated the clinical course of disease. NPC transplantation down-regulated the inflammatory brain process at the acute phase of disease, as indicated by a reduction in the number of perivascular infiltrates and of brain CD3+ T cells, an increase in the number and proportion of regulatory T cells and a reduction in the expression of ICAM-1 and LFA-1 in the brain. Demyelination and acute axonal injury in this model are considered to result mainly from the acute inflammatory process and correlate well with the chronic neurological residua. In consequence to inhibition of brain inflammation, precursor cell transplantation attenuated the primary demyelinating process and reduced the acute axonal injury. As a result, the size of demyelinated areas and extent of chronic axonal pathology were reduced in the transplanted brains. We suggest that the beneficial effect of transplanted NPCs in chronic EAE is mediated, in part, by decreasing brain inflammation and reducing tissue injury. PMID- 16472807 TI - Modulating the splicing activity of Tetrahymena ribozyme via RNA self-assembly. AB - The internal guiding sequence (IGS) is normally located at the 5' end of trans splicing ribozymes that are derived from the Tetrahymena group I intron, and is required for the recognition of substrate RNAs and for trans-splicing reactions. Here, we separated the Tetrahymena group I intron at the L2 loop to produce two fragments: the IGS-containing substrate, and the IGS-lacking ribozyme. We show here that two fragments can complex not through the IGS interaction but under the guidance of appended interacting nucleotides, and perform trans-splicing. The splicing reactions took place both in vitro and in mammalian cells, and the spliced mRNA product from the self-assembled ribozyme complex can be translated into functional proteins in vivo. The splicing efficiency was dependent on the length of appending nucleotides. PMID- 16472808 TI - Sequence-specific interference by small RNAs derived from adenovirus VAI RNA. AB - A virus-associated RNA (VAI) of adenoviruses is a cytoplasmic non-coding RNA and it plays an important role for viral replication in infected cells. VAI RNA transcripts, produced by RNA polymerase III (pol III), form tightly structured stems, which confer resistance to cellular defense systems. We demonstrate here that small RNAs of approximately 22 nucleotides are produced from a terminal stem region but not from an apical stem of VAI RNA. We determined the processing sites of VAI RNA by S1 nuclease mapping and further confirmed that the processed small RNA can act as small interfering RNAs (siRNAs) or as microRNAs (miRNAs) in transient transfection assays and during viral infection. Our data demonstrate that non-coding RNAs synthesized by pol III can be substrates for Dicer, and diced small RNAs might regulate cellular phenomena as siRNAs and miRNAs. PMID- 16472809 TI - Phylogeny of the corticotropin-releasing factor family of peptides in the metazoa. AB - The corticotropin-releasing factor (CRF) family of peptides consists of four distinct paralogs found on separate chromosomes in vertebrates. Among invertebrates, the family has been relatively well characterized in the insects where at least 2 or 3 paralogs, a CRF-binding protein ortholog and a CRF receptor variant have been found. The conservation of structure and function of this system in insects imply that the CRF system evolved in ancestral species well before the Precambrian explosion. The CRF family peptides association with diuresis and feeding may have developed early in its evolution. However, CRF's role in the hypothalamo-pituitary-adrenal axis and regulation of the glucocorticoids in association with energy metabolism appears to have developed in the chordate lineage. PMID- 16472810 TI - Hormonal regulation of ion and water transport in anuran amphibians. AB - Amphibians occupy a wide variety of ecological habitats, and their adaptation is made possible through the specialization of the epithelia of their osmoregulatory organs, such as the skin, kidney, and urinary bladder, which control the hydromineral and acid-base balance of their internal medium. Amphibians can change drastically plasma Na+, Cl-, and urea levels and excretion rates in response to environmental stimuli such as acute desiccation and changes in external salinity. Several hormones and the autonomic nervous system act to control osmoregulation. Several ion channels including an epithelial sodium channel (ENaC), a urea transporter (UT), and water channels (AQPs) are found in epithelial tissues of their osmoregulatory organs. This mini review examines the currents status of our knowledge about hormone receptors for arginine vasotocin, angiotensin II and aldosterone, and membrane ion channels and transporters, such as ENaC, UT, and AQPs in amphibians. PMID- 16472811 TI - Functional characterisation of the regulatory subunit of cyclic AMP-dependent protein kinase A homologue of Giardia lamblia: Differential expression of the regulatory and catalytic subunits during encystation. AB - To understand the functional roles of protein kinase A (PKA) during vegetative and differentiating states of Giardia parasites, we studied the structural and functional characteristics of the regulatory subunit of PKA (gPKAr) and its involvement in the giardial encystment process. Molecular cloning and characterisation showed that gPKAr contains two tandem 3'5'-cyclic adenosine monphosphate (cyclic AMP) binding domains at the C-terminal end and the interaction domain for the catalytic subunit. A number of consensus residues including in vivo phosphorylation site for PKAc and dimerisation/docking domain are present in gPKAr. The regulatory subunit physically interacts with the catalytic subunit and inhibits its kinase activity in the absence of cyclic AMP, which could be partially restored upon addition of cyclic AMP. Western blot analysis showed a marked reduction in the endogenous gPKAr concentration during differentiation of Giardia into cysts. An increased activity of gPKAc was also detected during encystation without any significant change in the protein concentration. Distinct localisations of gPKAc to the anterior flagella, basal bodies and caudal flagella as noted in trophozoites were absent in encysting cells at later stages. Instead, PKAc staining was punctate and located mostly to the cell periphery. Our study indicates possible enrichment of the active gPKAc during late stages of encystation, which may have implications in completion of the encystment process or priming of cysts for efficient excystation. PMID- 16472812 TI - Methodology to determine failure characteristics of planar soft tissues using a dynamic tensile test. AB - Predicting the injury risk in automotive collisions requires accurate knowledge of human tissues, more particularly their mechanical properties under dynamic loadings. The present methodology aims to determine the failure characteristics of planar soft tissues such as skin, hollow organs and large vessel walls. This consists of a dynamic tensile test, which implies high-testing velocities close to those in automotive collisions. To proceed, I-shaped tissue samples are subjected to dynamic tensile tests using a customized tensile device based on the drop test principle. Data acquisition has especially been adapted to heterogeneous and soft biological tissues given that standard measurement systems (considered to be global) have been completed with a non-contact and full-field strain measurement (considered to be local). This local measurement technique, called the Image Correlation Method (ICM) provides an accurate strain analysis by revealing strain concentrations and avoids damaging the tissue. The methodology has first been applied to human forehead skin and can be further expanded to other planar soft tissues. The failure characteristics for the skin in terms of ultimate stress are 3 MPa +/- 1.5 MPa. The ultimate global longitudinal strains are equal to 9.5%+/-1.9% (Green-Lagrange strain), which contrasts with the ultimate local longitudinal strain values of 24.0%+/-5.3% (Green-Lagrange strain). This difference is a consequence of the tissue heterogeneity, clearly illustrated by the heterogeneous distribution of the local strain field. All data will assist in developing the tissue constitutive law that will be implemented in finite element models. PMID- 16472813 TI - Application of internal universal calibration for determination of fully dissociated species in capillary electrophoresis using indirect UV detection and electrokinetic injection. AB - Although the electrokinetic injection suffers from quantitation problems arising from mobility and matrix bias, the application of electrokinetic injection and indirect photometric detection may provide a very simple, sensitive and universal determination of fully dissociated components. In this work a survey of the possibilities of quantitative analysis using EK injection with indirect UV detection is given and a simple procedure (internal universal calibration) for accurate quantitative determination of traces using EK injection is proposed. It was shown that the thiosulfate as an internal universal standard (IUS) can be applied for quantitative determination of fully dissociated analytes (e.g. Br(-), Cl(-), NO(2)(-), NO(3)(-), SO(4)(2-), S(2)O(3)(2-)) whose mobilities (migration times) do not differ from the mobilities of monitoring ion (chromate), counter ion (potassium) and IUS with more than 10%. The proposed calibration procedure provides concentration data, which deviate generally less than 5% extent from the actual true values. The validity and the applicability of the method was studied in the analysis of samples containing different amount of matrix material. PMID- 16472814 TI - High-performance liquid chromatographic method for determination of reversible isomers of SU5416. AB - SU5416 shows light-induced reversible geometric isomerism. A simple, reliable, isocratic HPLC method using an UV-vis detector at lambda(425nm) was developed. The method provides efficient (R(S)=3.5) analysis of the two isomers with retention of the isomeric integrity. Additionally, the method has linearity over a wide range (50-1000microg/mL, r(2)=0.99), is accurate (99-102%, RSD <4%), and reproducible (RSD <0.8%). The method was used for analyzing pharmaceutical samples and understanding the kinetics of SU5416 isomers in methanol. In addition, this method can be used for quantifying the non-isolatable E-isomer. PMID- 16472815 TI - Atmospheric pressure photoionization mass spectrometry for analysis of fatty acid and acylglycerol lipids. AB - In this work, we optimize parameters and conditions for analysis of fatty acid ester and acylglycerol lipids by atmospheric pressure photoionization-mass spectrometry (APPI-MS). The investigated parameters include atmospheric pressure chemical ionization (APCI) nebulizer/vaporizer physical orientation and APPI lamp face position, solvent selections, mobile phase compositions and flow rates, cone voltages and probe temperatures. APPI sensitivity is found to be highly dependent on mobile phase compositions. Normal phase solvents offer much higher sensitivity and better peak shape than reversed phase for nonpolar lipids. Hexane and isooctane are found to be two solvents generating highest S/N for eicosapentaenoic acid (EPA) methyl ester. The effects of mobile phase flow rates on sensitivity are found to be target analytes and target ions specific. However, the flow rate changes do not significantly affect the sensitivity of three out of four tested analytes under normal phase conditions over tested flow rates of 50 500muL/min. Cone voltage is found to be one of key parameters affecting sensitivity. Optimum probe temperature is found to be more dependent on mobile phase compositions than on the specific target analytes. Aqueous reversed-phase mobile phase requires higher probe temperature than normal phase for better sensitivity. More volatile mobile phase solvents require lower probe temperature for analyte desolvation. APPI offers four to five decades of linear ranges under normal phase condition. Full scan mass spectra of individual lipid standards, custom lipid mixtures and natural fish oil show that APPI spectra are clean and very easy to interpret. APPI also gives stable, reproducible peak responses with good peak shape. Limits of detection (LODs) by FIA (S/N=3) are estimated to be 12pg for EPA methyl ester and monoarachidin, 19pg for diarachidin and 7pg for trielaidin. LODs on-column are estimated to be 94pg for EPA methyl ester, 90pg for monoarachidin and diarachidin and 24pg for trielaidin. PMID- 16472816 TI - Two-dimensional gas chromatography coupled to triple quadrupole mass spectrometry for the unambiguous determination of atropisomeric polychlorinated biphenyls in environmental samples. AB - Two-dimensional gas chromatography coupled to triple quadrupole mass spectrometry (GC-GC-MS-MS) is presented as a robust, selective and sensitive method for the accurate quantification of enantiomeric fractions (EFs) of atropisomeric polychlorinated biphenyls (PCBs) in a range of environmental matrices. The method was subjected to a set of quality control and validation experiments. EF precision of the atropisomers 95, 149, 132, and 174 ranged from 0.7 to 1.4% in soil samples. Quantified amounts of these congeners in soil and compost samples (59T:A transversions, a signature mutation of oxidative DNA damage. Ercc1-/m mice, with their short lifespan of 6 months and severe symptoms of premature aging, especially in liver and kidney, displayed an even faster lacZ-mutant accumulation in liver. In this case, the excess mutations were mostly genome rearrangements. Csb-/- mice, with mild premature aging features and no reduction in lifespan, and Xpdm/m mice, exhibiting prominent premature aging features and about 20% reduction in lifespan, did not have elevated lacZ-mutant frequencies. It is concluded that while increased genomic instability could play a causal role in the mildly accelerated aging phenotype in the Xpa-null mice or in the severe progeroid symptoms of the Ercc1-mutant mice, shortened lifespan in mice with defects in transcription-related repair do not depend upon increased mutation accumulation. PMID- 16472828 TI - Exogenous 8-oxo-dG is not utilized for nucleotide synthesis but enhances the accumulation of 8-oxo-Gua in DNA through error-prone DNA synthesis. AB - 7,8-Dihydro-8-oxoguanine (8-oxo-Gua) and its nucleoside in cytosol are derived from the repair of oxidative DNA and the cleanup of oxidatively damaged DNA precursors, respectively. While the harmful effects of 8-oxo-Gua present in DNA have been studied extensively, few have reported its effects on cytosolic function. Our previous study showed that the addition of 8-oxo-dG to culture media caused an accumulation of 8-oxo-Gua in nuclear DNA in several leukemic cells including KG-1, which lack 8-oxoguanine glycosylase 1 (OGG1) activity due to mutational loss. However, the mechanism underlying 8-oxo-Gua level increases in DNA has not been addressed. In this study, we elucidated the metabolic fate of 8-oxo-Gua-containing nucleotide and the effect of exogenous 8-oxo-dG on DNA synthesis in KG-1 cells. We found that 8-oxo-dGMP was rapidly dephosphorylated to 8-oxo-dG rather than phosphorylated to 8-oxo-dGDP, thus indicating that 8-oxo-Gua containing molecule is not used as a substrate for DNA synthesis in KG-1 cells. In fact, radiolabeled 8-oxo-dG was incubated but radioactivity was not detected in nuclear DNA of KG-1 cells, showing that 8-oxo-dG is not directly incorporated into DNA. Interestingly, the activity of DNA polymerase beta, which synthesize DNA with low fidelity increased in KG-1 cells treated with 8-oxo-dG, whereas the expression of DNA polymerase alpha decreased. In addition, the accumulation of 8 oxo-Gua in KG-1 DNA was completely inhibited by a specific inhibitor of DNA polymerase beta. Thus, our findings address that the insertion of 8-oxo-dG into KG-1 DNA is not due to the direct incorporation of exogenous 8-oxo-dG, but rather to the inaccurate incorporation of endogenous 8-oxo-dGTP by DNA polymerase beta. It further suggests that 8-oxo-dG in the cytosol may function as an active molecule itself and perturb the well-defined DNA synthesis. PMID- 16472829 TI - Replication restart: a pathway for (CTG).(CAG) repeat deletion in Escherichia coli. AB - (CTG)n.(CAG)n repeats undergo deletion at a high rate in plasmids in Escherichia coli in a process that involves RecA and RecB. In addition, DNA replication fork progression can be blocked during synthesis of (CTG)n.(CAG)n repeats. Replication forks stalled at (CTG)n.(CAG)n repeats may be rescued by replication restart that involves recombination as well as enzymes involved in replication and DNA repair, and this process may be responsible for the high rate of repeat deletion in E. coli. To test this hypothesis (CAG)n.(CTG)n deletion rates were measured in several E. coli strains carrying mutations involved in replication restart. (CAG)n.(CTG)n deletion rates were decreased, relative to the rates in wild type cells, in strains containing mutations in priA, recG, ruvAB, and recO. Mutations in priB and priC resulted in small reductions in deletion rates. In a recF strain, rates were decreased when (CAG)n comprised the leading template strand, but rates were increased when (CTG)n comprised the leading template. Deletion rates were increased slightly in a recJ strain. The mutational spectra for most mutant strains were altered relative to those in parental strains. In addition, purified PriA and RecG proteins showed unexpected binding to single-stranded, duplex, and forked DNAs containing (CAG)n and/or (CTG)n loop-outs in various positions. The results presented are consistent with an interpretation that the high rates of trinucleotide repeat instability observed in E. coli result from the attempted restart of replication forks stalled at (CAG)n.(CTG)n repeats. PMID- 16472830 TI - Identification of somatic and germline mitochondrial DNA sequence variants in prostate cancer patients. AB - Prostate cancer (PCa) is the second most frequent cancer among men in the European Union and the most common in the United States. Older age and a positive family history of PCa are important risk factors, but little is known about the disease aetiology. Mitochondria are involved in essential cellular pathways, some of which have been associated with tumorigenesis. We analysed the presence of sequence variants, depletion and rearrangements in the mitochondrial DNA (mtDNA) of PCa patients. Sequencing of the D-loop and genes RNR1 and 2, ND3, ND4L and ND4, and long-range and real-time PCR techniques were carried out on 51 samples (41 from patients and 10 from controls). Normal, hyperplastic or tumour samples were obtained from 17 patients. Six patients' seminal vesicles were also investigated as an additional patient's control tissue (these structures seldom develop tumours). Neither depletion nor mtDNA rearrangements were detected. In contrast, 94 mtDNA sequence variants were identified, 9 previously unreported. The regions presenting more sequence variants were MT-DLOOP (52%), MT-RNR2 (14%) and MT-ND4 (13%). The patients' seminal vesicles studied showed the same set of variants as the corresponding prostate, suggesting either that the pathogenic role of these particular variants is minor or that they participate in the prostatic carcinogenesis in combination with other factors absent in seminal vesicles. Five patients (29.4%) harboured eight somatic changes in the mtDNA. One affects a conserved residue and three have not been previously described. The analysis of other genes in the mtDNA molecule might demonstrate an even higher incidence of mtDNA somatic variants in these PCa patients. PMID- 16472831 TI - The role of DNA polymerase iota in UV mutational spectra. AB - UVB (280-320 nm) and UVC (200-280 nm) irradiation generate predominantly cyclobutane pyrimidine dimers (CPDs) and (6-4) photoproducts in DNA. CPDs are thought to be responsible for most of the UV-induced mutations. Thymine-thymine CPDs, and probably also CPDs containing cytosine, are replicated in vivo in a largely accurate manner by a DNA polymerase eta (Pol eta) dependent process. Pol eta is a DNA damage-tolerant and error-prone DNA polymerase encoded by the POLH (XPV) gene in humans. Another member of the Y family of error-prone DNA polymerases is POLI encoding DNA polymerase iota (Pol iota). In order to clarify the specific role of Pol iota in UV mutagenesis, we have used an siRNA knockdown approach in combination with a supF shuttle vector which replicates in mammalian cells, similar as we have previously done for Pol eta. Synthetic RNA duplexes were used to efficiently inhibit Pol iota expression in 293 T cells. The supF shuttle vector was irradiated with 254 nm UVC and replicated in 293 T cells in presence of anti-Pol iota siRNA. Surprisingly, there was a consistent reduction of recovered plasmid from cells with Pol iota knockdown and this was independent of UV irradiation of the plasmid. The supF mutant frequency was unchanged in the siRNA knockdown cells relative to control cells confirming that Pol iota does not play an important role in UV mutagenesis. UV-induced supF mutants were sequenced from siRNA-treated cells and controls. Neither the type of mutations nor their distribution along the supF gene were significantly different between controls and siRNA knockdown cells and were predominantly C to T and CC to TT transitions at dipyrimidine sites. These results show that Pol iota has no significant role in UV lesion bypass and mutagenesis in vivo and provides some initial data suggesting that this polymerase may be involved in replication of extrachromosomal DNA. PMID- 16472832 TI - Partial lesion of the serotonergic system by a single dose of MDMA results in behavioural disinhibition and enhances acute MDMA-induced social behaviour on the social interaction test. AB - The acute effects of 3,4-methylenedioxymethamphetamine (MDMA, ecstasy) on anxiety related behaviours were studied using indices of social interaction in Dark Agouti (DA) both drug naive rats and those pretreated with MDMA (15 mg/kg i.p.) 3 weeks earlier. The functional neuroanatomy of these MDMA effects was visualised using 2-deoxyglucose imaging of local cerebral glucose use (LCMRglu), whilst MDMA induced serotonergic neurotoxicity was measured by radioligand binding with [3H]paroxetine. Acute MDMA alone markedly decreased most typical elements of social interaction but increased adjacent lying, a behaviour that also contains social elements. In animals pre-exposed to MDMA, decreased [3H]paroxetine binding indicated serotonergic terminal depletion, and in these animals significant increases in locomotor activity, exploratory behaviour and aggressive behaviour were found. Both behavioural effects and also the metabolic activation induced by acute MDMA were potentiated in rats previously exposed to the drug. In conclusion, a single dose of MDMA caused marked changes in social behaviour acutely that might be interpreted either as a decrease or increase in anxiety. Three weeks after MDMA a behavioural disinhibition similar to psychomotor agitation, a symptom connected to depression or mania, and a sensitization to the acute effects of MDMA are apparent in both the behavioural and brain metabolic effects of the drug. PMID- 16472833 TI - Rapid turnover and polyubiquitylation of the retroviral restriction factor TRIM5. AB - TRIM5alpha and TRIMCyp are retroviral restriction factors that, like other members of the tripartite motif (TRIM) family, contain RING, B-box 2 and coiled coil domains. We found that both proteins are rapidly turned over, with half lives of 50-60 min. Polyubiquitylation and rapid degradation of TRIM5alpha depended upon intact RING and B-box 2 domains. A chimera consisting of monkey TRIM5alpha with a RING domain of human TRIM21 exhibited a half-life of 210 min, yet potently restricted human immunodeficiency virus; therefore, rapid turnover of TRIM5alpha is not required for its antiretroviral activity. TRIM5alpha forms cytoplasmic bodies that contain other polyubiquitylated proteins, heat shock proteins and dynein, and thus resemble aggresome precursors. Consistent with this interpretation, proteasomal inhibitors triggered the formation of TRIM5alpha(rh) containing aggresomes in a microtubule-dependent manner. Thus, TRIM5alpha levels in the cell are maintained by continuous synthesis and rapid proteasome-mediated degradation, imbalances in which result in the formation of pre-aggresomal cytoplasmic bodies. PMID- 16472834 TI - Downregulation of human immunodeficiency virus type 1 Gag expression by a gp41 cytoplasmic domain fusion protein. AB - The cytoplasmic domain of human immunodeficiency virus type 1 (HIV-1) envelope (Env) transmembrane protein gp41 interacts with the viral matrix MA protein, which facilitates incorporation of the trimeric Env complex into the virus. It is thus feasible to design an anti-HIV strategy targeting this interaction. We herein describe that Gag expression can be downregulated by a cytoplasmic domain fusion protein of the Env transmembrane protein, beta-galactosidase (beta gal)/706-856, which contains the cytoplasmic tail of gp41 fused at the C terminus of Escherichia coli beta-gal. This mediator depleted intracellular Gag molecules in a dose-dependent manner. Sucrose gradient ultracentrifugation and confocal microscopy revealed that Gag and beta-gal/706-856 had stable interactions and formed aggregated complexes in perinuclear, intracellular sites. Pulse-chase and cycloheximide chase analyses demonstrated that this mediator enhanced unmyristylated Gag degradation. The results demonstrate a novel mode of HIV-1 Gag downregulation by directing Gag to an intracellular site via the interaction of Gag with a gp41 cytoplasmic domain fusion protein. PMID- 16472835 TI - Methylation of the human papillomavirus-18 L1 gene: a biomarker of neoplastic progression? AB - Epigenetic transcriptional regulation plays an important role in the life cycle of human papillomaviruses (HPVs) and the carcinogenic progression of anogenital HPV associated lesions. We performed a study designed to assess the methylation status of the HPV-18 genome, specifically of the late L1 gene, the adjacent long control region (LCR), and part of the E6 oncogene in cervical specimens with a range of pathological diagnoses. In asymptomatic infections and infections with precancerous (precursor) lesions, HPV-18 DNA was mostly unmethylated, with the exception of four samples where hypermethylation of L1 was detected. In contrast, L1 sequences were strongly methylated in all cervical carcinomas, while the LCR and E6 remained unmethylated. HeLa cells, derived from a cervical adenocarcinoma, contain chromosomally integrated HPV-18 genomes. We found that L1 is hypermethylated in these cells, while the LCR and E6 are unmethylated. Treatment of HeLa cells with the methylation inhibitor 5-Aza-2'-deoxycytidine (5-Aza-CdR) led to the expected reduction of L1 methylation. After removal of 5-Aza-CdR, L1 methylation resumed and exceeded pretreatment levels. Unexpectedly, the LCR and E6 also became methylated under these conditions, albeit at lower levels than L1. We hypothesize that L1 is preferentially methylated after integration of the HPV genome into the cellular DNA, possibly since linearization prohibits its normal transcription, while the enhancer and promoter may be protected from methylation by transcription factors. Since our data suggest that HPV-18 L1 methylation can only be detected in carcinomas, except in some few precancerous lesions and asymptomatic infections, L1 methylation may constitute a powerful molecular marker for detecting this important step of neoplastic progression. PMID- 16472836 TI - Amodal completion impairs stereoacuity discrimination. AB - Visual stimulus configuration can influence elementary visual processes. We provide empirical evidence to demonstrate this effect in stereoscopic depth discrimination. Two vertically aligned bars were presented in stereo such that one of them was closer to the human observer. Observers discriminated which of the two was closest. In the first, "occluded" condition, a horizontal bar, positioned closest in depth to the observer, was added to the display such that the two vertical bars perceptually completed to form a whole by connecting together behind the horizontal bar. In the second, control condition, the horizontal bar was placed furthest away from the observer such that there was a visible gap between the two vertical bars, which could no longer complete perceptually. We measured observers' psychometric functions using the method of constant stimuli, and found that their discrimination sensitivity d' was smaller when the two vertical bars perceptually completed than when they did not. We used a simple model to illustrate that when the two vertical bars perceptually completed, they also tended to be perceived as coplanar in the fronto-parallel plane. This consequence of completion made it more difficult to discriminate any difference in depth between the two vertical bars. PMID- 16472838 TI - Removal of methylene blue from waste water using micellar enhanced ultrafiltration. AB - Ultrafiltration of micellar solutions containing sodium dodecylsulfate (SDS) and oxyethylated coconut fatty acid methyl esters (OMC-10) and their binary mixtures were studied and used to recover methylene blue. This was achieved through solubilization in mixed negatively charged micelles. Under the experimental conditions used, no significant fouling of the membrane comprising of cellulose, polyethersulfone and polyvinylidene fluoride was observed, with no retardation of ultrafiltration. The introduction of a nonionic surfactant to SDS reduced the critical micelle concentration of mixed micelles and then the concentration of surfactants in the permeate, i.e., from 8.3 x 10(-3)M for SDS to 2.0 x 10(-3) M for the mixture of SDS:OMC-10 = 4:1. Such a tailored surfactant mixture enabled the authors to achieve 93-94% retention of methylene blue using a hydrophilic membrane made of cellulose. Ultrafiltration of micellar solutions could also be considered a research method, helpful in determining important parameters such as micelle loading and the micelle binding constant used to characterize micellar solutions. Additionally the distribution coefficient employed in extraction studies can be resolved. PMID- 16472837 TI - Macular pigment and ocular biometry. AB - This study is designed to investigate the relationship between macular pigment optical density (MPOD) and ocular biometric parameters. The following details were recorded for 180 healthy subjects: demographic profile; best-corrected visual acuity; refractive status; ocular biometric parameters [axial length (AL), anterior chamber depth (ACD), lens thickness (LT) and vitreous chamber depth (VCD)]; ocular dominance; MPOD; serum lutein (L) and zeaxanthin (Z). The mean MPOD (+/-SD) was 0.307 (0.155) and 0.305 (0.149) in the right and left eyes, respectively. No demonstrable relationship was observed between MPOD and AL, ACD or VCD [AL: r=0.091, p=0.225; ACD: r=0.091, p=0.227; VCD: r=0.146, p=0.051]. There was a significant and inverse relationship between LT and MPOD (r=-0.204; p=0.008), which was attenuated to non-significance after correction for age and height (r=-0.058; p=0.466). This study fails to identify an association between MPOD and ocular biometric parameters. This is an important negative finding, which allows investigators to study MP, and its relationship with potentially important variables, without the need to correct for ocular biometric parameters. PMID- 16472839 TI - Hominin lower second premolar morphology: evolutionary inferences through geometric morphometric analysis. AB - Mandibular premolars are increasingly used in taxon-specific diagnostic analyses of hominins. Among the principal difficulties in these evaluations is the absence of discrete, discernible, and comparable anatomical structures for rigorous quantitative assessment. Previous research has addressed either internal crown surface features (such as cusps and fossae) or the morphology of the crown outline. In the present paper, we integrate both types of information in the examination of morphological variation of lower P4s (n = 96) among various fossil hominin species with an emphasis on genus Homo. We use a set of 34 2D landmarks combining coordinate data from four classical dental landmarks on the occlusal surface and 30 sliding semilandmarks of the crown outline. Our results indicate that external shape variation is closely related to the configuration of the occlusal morphological features and influenced by dental size. The external and internal shapes of P4 are polymorphic but still useful in depicting a primitive derived gradient. The primitive pattern seems to have been an asymmetrical contour with a mesially displaced metaconid, development of a bulging talonid, and a broad occlusal polygon. The trend toward dental reduction during the Pleistocene produced different morphological variants with a reduced occlusal polygon and decreased lingual occlusal surface in later Homo species. Homo heidelbergensis/neanderthalensis have fixed plesiomorphic traits in high percentages, whereas in modern humans a symmetrical outline with a centered metaconid and talonid reduction evolved. PMID- 16472840 TI - Population history, biogeography, and taxonomy of orangutans (Genus: Pongo) based on a population genetic meta-analysis of multiple loci. AB - This paper examines orangutan population history and evolution through a meta analysis of seven loci collected from both Sumatran and Bornean orangutans. Within orangutans, most loci show that the Sumatran population is about twice as diverse as the Bornean population. Orangutans are more diverse than African apes and humans. Sumatran and Bornean populations show significant genetic differentiation from one another and their history does not differ significantly from an 'island model' (population splitting without gene flow). Two different methods support a divergence of Bornean and Sumatran orangutans at 2.7-5 million years ago. This suggests that Pleistocene events, such as the cyclical exposure of the Sunda shelf and the Toba volcanic eruption, did not have a major impact on the divergence of Bornean and Sumatran orangutans. Pairwise mismatch analyses, however, suggest that Bornean orangutans have undergone a recent population expansion (beginning 39,000-64,000 years ago), while Sumatran orangutan populations were stable. Pleistocene events may have contributed to these aspects of orangutan population history. These conclusions are applied to the debate on orangutan taxonomy. PMID- 16472841 TI - A new pliopithecoid genus from the early Miocene of Uganda. AB - A partial face and mandible from the early Miocene site of Napak IX in Uganda are described here as a new genus and species of catarrhine primate, Lomorupithecus harrisoni gen. et sp. nov. The face is among the most complete specimens known for a Miocene small-bodied catarrhine. Several aspects of its anatomy indicate that the new species is a stem catarrhine, and as such, it may provide valuable information pertaining to the primitive catarrhine cranial morphotype. Lomorupithecus is most similar in its facial anatomy to members of the Pliopithecoidea, and these similarities could be interpreted in three ways. They could be symplesiomorphies, which would support the traditional view of the primitive catarrhine cranial morphotype; they could be synapomorphies reflecting a phylogenetic position of Lomorupithecus within Pliopithecoidea; or they could represent convergence. Phylogenetic analysis of Lomorupithecus along with 35 other primates indicates that it is a pliopithecoid. As such, it would be the oldest and only Afro-Arabian member of this otherwise Eurasian clade. PMID- 16472842 TI - Sequence variations within the genes related to hemostatic imbalance and their impact on coronary artery disease in Turkish population. AB - Coronary artery disease (CAD) is reported to be associated with some genetic risk factors. Since identification of genetic risk factors for CAD in different ethnic groups is important for the development of new intervention and prevention programs, we investigated the association between the R353Q and -323ins10 polymorphisms in Factor VII gene, C677T mutation in MTHFR, Factor V Leiden and PT G20210A mutations and CAD in Turkish population. The promoter region of the PAI-1 gene was also screened by SSCA (single-stranded conformation analysis) using specifically designed primers. 137 CAD patients with early onset documented by coronary angiography and 41 individuals who had no significant coronary stenosis by angiography as control group were screened for the identification of the polymorphisms. In conclusion, Factor V Leiden was found to be an independent genetic risk factor for CAD in Turkish population. Combined risk assessment indicated that the coexistence of two other inherited thrombophilia markers, namely MTHFR C677T and PT G20210A with Factor V Leiden may increase the risk of the development of the disease in this population. The results of the present study show that there is no statistically significant association between the two polymorphisms in Factor VII gene, MTHFR C677T polymorphism, PT G20210A polymorphism, 4G/5G polymorphism of PAI-1 and CAD in Turkish population. PMID- 16472843 TI - Type I and type III collagen-platelet interaction: inhibition by type specific receptor peptides. AB - We have previously cloned and characterized a platelet receptor for type III collagen (47 kDa) from a human bone marrow cDNA phage library and defined two active peptides. We also cloned and characterized a platelet receptor for type I collagen (65 kDa) and defined an active peptide. Our objective was to study whether there is type specificity of these active peptides. We have engineered a mutant receptor clone by replacing one of the two active peptides of the platelet receptor for type III collagen with the active peptide of the platelet receptor for type I collagen. The replacement of an active peptide at the amino terminal end (rMIII) of the platelet receptor for type III collagen with the type I collagen active peptide was done without altering the hydrophilicity of the protein. This purified recombinant protein reacts with polyclonal anti-47-kDa and anti-65-kDa active peptide antibodies. The purified recombinant protein inhibits both types I and III collagen-induced platelet aggregation. This rMIII also inhibits the adhesion of washed platelets to rabbit aortic segments (natural matrix) in a dose-dependent manner. The chemically synthesized hybrid peptide of each active peptide of platelet type I and type III collagen receptors inhibits types I and III collagen-induced platelet aggregation in a dose-dependent manner. These results suggest that there is a type specific reactive site on platelets for type I and type III collagens. PMID- 16472844 TI - Total laparoscopic radical hysterectomy and lymphadenectomy: the M. D. Anderson Cancer Center experience. AB - OBJECTIVE: To retrospectively evaluate the feasibility and morbidity of total laparoscopic radical hysterectomy and lymphadenectomy. METHODS: We performed a retrospective review of all patients who underwent a total laparoscopic radical hysterectomy at our institution between May 2004 and August 2005. Data collected included age, body mass index, stage, histopathologic subtype, tumor grade, estimated blood loss, perioperative blood transfusions, number and status of lymph nodes obtained, status of surgical margins, length of hospital stay, time to resumption of normal bladder function, intraoperative and postoperative complications, and disease-free interval. RESULTS: Twenty patients underwent total laparoscopic radical hysterectomy during the study period. None of the surgeries required conversion to laparotomy. The median patient age was 41.5 years (range, 25-76). Eighteen patients had cervix cancer (5 stage IA2 and 13 stage IB1), and 2 had endometrial cancer (1 stage IB and 1 stage IIIA). Among those with cervix cancer, 12 had adenocarcinoma, 4 squamous cell carcinoma, and 2 adenosquamous carcinoma. The median weight was 70 kg (range, 49-112). The median number of resected pelvic lymph nodes was 13 (range, 9-26). One patient had nodal disease. The surgical margins were free of disease in all cases. The median blood loss was 200 ml (range, 25-700 ml). Only 1 patient required an intraoperative blood transfusion (1 U packed red blood cells). The median length of hospital stay was 1 day (range, 1-5). There were 3 short-term complications--unintentional cystotomy, pulmonary embolus, and pneumomediastinum with subcutaneous emphysema. There were 2 long-term complications--vaginal eviscerations and a lymphocyst. The median time to resumption of normal bladder function was 16 days (range, 13-29). The median follow-up time was 8 months range (1-16). All patients remain free of disease at the time of this report. CONCLUSIONS: Total laparoscopic radical hysterectomy can be performed safely with minimal blood loss and postoperative morbidity, and patients undergoing this procedure may be discharged after an overnight stay in the hospital. PMID- 16472845 TI - Langerhans' cells and smoking in intraepithelial neoplasia of the cervix. AB - OBJECTIVE: To compare the intraepithelial population of Langerhans' cells (LC) in normal cervix epithelium adjacent to cervical intraepithelial neoplasia grade 3 (CIN 3) and correlate to smoking habit. METHODS: Cases in this study included conization specimens from 48 women affected by CIN 3. The LC count was performed in areas without histopathologic alteration adjacent to CIN 3. The control group is compound by normal cervix from 46 hysterectomy specimens. The identification of LC was done by immunohistochemical study demonstrating immunoreactivity to S 100 protein. The number of intraepithelial LC was determined using 400x magnification light microscope in 10 high-power fields, and results were expressed in number of cells per square millimeter (LC/mm(2)). RESULTS: In the control group, there was lower number of Langerhans' cells in smokers than in non smokers (P = 0.045). There was lower number of Langerhans' cells in normal areas adjacent to CIN 3 than in normal cervix control group (P = 0.004). There was no significant difference in the number of Langerhans' cells in normal areas of the cervix with CIN 3 between smokers and non-smokers (P = 0.991). The number of cigarettes consumed daily, time of consume, total number of cigarettes consumed showed a reduced LC count, yet was not statistically significant. CONCLUSIONS: It was revealed that smoking reduces the number of intraepithelial Langerhans' cells in the uterine cervix. In women with CIN 3, the LC count was lower, despite the habit of smoking. PMID- 16472846 TI - Re: "Is the assumed natural history of vulvar intraepithelial neoplasia III based on enough evidence? A systematic review of 3322 published patients". PMID- 16472847 TI - Endocervical glandular involvement is associated with high-grade SIL. AB - OBJECTIVE: To examine whether endocervical glandular involvement (EGI) by squamous intraepithelial lesions (SIL) is associated with the grade of the squamous lesion and to assess the usefulness of such association for diagnostic purposes. METHODS: Histologic sections from two hundred successive cases of cervical SIL were reviewed retrospectively from the archives of the Department of Pathology of a tertiary care medical center. RESULTS: Nine of 93 (9.7%) low-grade lesions and 41 of 107 (38.3%) high-grade lesions showed glandular involvement (P = 0.001, Chi-squared test). The relative risk of having EGI in high-grade lesions as compared to low-grade lesions was 3.95. CONCLUSION: EGI is seen approximately four times more frequently in high-grade SIL than in low-grade SIL. Therefore, in cases that are difficult to grade, the presence of EGI is in favor of the lesion being a high-grade SIL. PMID- 16472848 TI - Effect of storage media and storage time on survival of spermatozoa recovered from canine and feline epididymides. AB - The aim of this study was evaluate the survival ability of canine and feline spermatozoa maintained within epididymides stored at 4 degrees C for 24, 48 or 72 h in sterile isotonic saline solution (SAL) or a Tris-egg yolk (TEY) storage medium. Fifteen domestic dogs and 15 cats were neutered and their testes were placed in TEY or SAL and stored at 4 degrees C for either 24, 48 or 72 h. Sperm samples were obtained by cutting the cauda epididymides into a Tris extender and were evaluated for motility, velocity, viability, plasma membrane integrity, and acrosome morphology. In dogs, there were no significant differences between storage media for motility, plasma membrane integrity, viability and velocity. However, dog sperm stored in TEY had better acrosome morphology compared to sperm stored in SAL (P < 0.05). Dog sperm recovered at 72 h had a reduction in all parameters studied compared to those recovered at 24 h (P < 0.05). In cats, sperm recovered from epididymides stored in TEY had higher motility, plasma membrane integrity and velocity at all times compared to those stored in SAL (P < 0.05). Cat sperm recovered at 72 h had reduced motility, acrosome morphology, viability and velocity compared to those recovered at 24 h (P < 0.05). The addition of TEY to canine epididymal sperm, thus, had a better protective effect than SAL only on acrosome morphology. In cats, in contrast, TEY had a better protective effect than SAL on all epididymal sperm parameters studied. In both species, sperm recovered at 72 h had a significant reduction in all parameters studied compared to those recovered at 24 h. PMID- 16472849 TI - Influence of cooling rates and addition of Equex pasta on cooled and frozen thawed semen of generic gray (Canis lupus) and Mexican gray wolves (C. l. baileyi). AB - A current priority for the preservation of the endangered Mexican gray wolf (Canis lupus baileyi) is the development of a sperm-based genome resource bank for subsequent use in artificial insemination. To optimize the quality of cryopreserved sperm, the procedures involved in processing semen before and during freezing need to be improved. The aim of this study were to examine the effects of: (i) different cooling periods before freezing and (ii) addition of Equex pasta (Minitub, Tubingen, Germany) on the characteristics of sperm from the generic gray wolf and the Mexican gray wolf after cooling and cryopreservation. For Mexican wolf sperm, cooling for 0.5 and 1.0 h had a less detrimental effect on cell morphology than cooling for 2.5 h, whereas the slower cooling rate (2.5 h) had a less detrimental effect on functional parameters and seemed to cause less damage to plasma membrane and acrosome integrity than 0.5 and 1.0 h. For the generic gray wolf, cooling semen for 2.5 h had less detrimental effect on plasma membrane integrity and viability; together with the 0.5 h cooling time, it yielded the highest percentages of intact acrosomes. As previously shown in the domestic dog, Equex pasta had no beneficial effect on sperm characteristics in either wolf species. PMID- 16472850 TI - Freezing of epididymal spermatozoa from dogs after cool storage for 2 or 4 days. AB - An experiment was conducted to investigate the freezing ability of canine epididymal spermatozoa after cool storage at 5 degrees C for 2 or 4 days. Spermatozoa were collected from the caudae epididymidis from 16 dogs. Total motility, plasma membrane integrity and acrosome integrity were evaluated immediately on harvesting, and after 2 and 4 days of storage at 5 degrees C, and at 0 and 2 h post-thaw at 37 degrees C. Sperm motility decreased significantly during cold storage, compared to freshly harvested spermatozoa (P < 0.001). Although there was no significant effect of pre-freeze storage time on post-thaw motility, there was a tendency towards decreased motility in spermatozoa that had been stored for 4 days, compared to spermatozoa that were frozen immediately after collection (P = 0.09). The number of post-thaw spermatozoa with an intact plasma membrane was decreased in spermatozoa cold-stored for 4 days (P < 0.001). There was no significant effect of pre-freeze storage time on the acrosomal status of post-thaw spermatozoa. In conclusion, canine epididymal spermatozoa were stored at 5 degrees C for up to 4 days without a clear detrimental effect on post-thaw motility and acrosome integrity, but storage may have decreased post thaw motility. Results were, however, generally low. PMID- 16472851 TI - Embryotrophic effects of ethylenediaminetetraacetic acid and hemoglobin on in vitro porcine embryos development. AB - This study investigated the embryotrophic effects of ethylenediaminetetraacetic acid (EDTA) and hemoglobin (Hb) on porcine preimplantation embryo development. Porcine embryos produced by in vitro maturation/fertilization were cultured for 6 days in modified North Carolina State University-23 medium (mNCSU-23) supplemented with EDTA and/or Hb. In Exp. 1, culturing porcine zygotes with 100 microM EDTA significantly increased cleavage frequencies (85.3%) at 48 h post insemination and the number of inner cell mass (ICM) (9.6+/-5.5) compared to the control (7.0+/-2.8). However, 100 microM EDTA did not improve blastocyst formation compared to 0, 1 or 10 microM EDTA. In Exp. 2, in vitro fertilized oocytes were cultured with 0, 1 or 10 microg/ml Hb. Culturing with Hb did not promote porcine embryo development, but significantly increased the cell numbers of blastocysts in 1 microg/ml Hb compared to 0 or 10 microg/ml Hb. In Exp. 3, culturing embryos with 100 microM EDTA+1 microg/ml Hb significantly improved frequencies of cleavage, blastocyst formation, and total cell numbers in blastocysts compared to the control. Moreover, 100 microM EDTA, 1 microg/ml Hb and their combination reduced reactive oxygen species (ROS) accumulation and decreased the incidence of apoptosis. In conclusion, the present study clearly demonstrated that the combining treatment of EDTA and Hb improved IVF porcine embryo development. PMID- 16472852 TI - Follicle-stimulating hormone receptor in felids: intra- and interspecies variation. AB - Assisted reproductive technologies are increasingly applied to support breeding efforts for many endangered felids. To explain the highly variable responses among felids to exogenous gonadotropins (FSH, eCG), we analyzed a 567bp fragment spanning a hyper-variable region of the FSH receptor in the domestic cat (catFSHR) and nine wild felid species/subspecies (felFSHR). Phylogenetic analysis indicated that the newly sequenced felFSHRs, together with the bear FSHR, belong to the carnivore group closely related to the ungulate clade. Within Felidae, genetic distances were 0.0089 +/- 0.0018 for nucleotide and 0.0183 +/- 0.0044 for amino acid (aa) sequences. In pairwise comparisons among catFSHR and all new felFSHRs, similarity ranged from 98.6 to 99.5% for nucleotides and from 97.4 to 98.9% for aa. Besides interspecies variability, intraspecies variation was also detected on both the cDNA and the protein level. There were no indications for an expression of tissue-specific isoforms of FSHR in testis and ovary. PMID- 16472853 TI - Ovarian response to repeated administration of alternating exogenous gonadotropin regimens in the ocelot (Leopardus pardalis) and tigrinus (Leopardus tigrinus). AB - Exogenous gonadotropins are used to stimulate ovarian follicular growth and ovulation in mammalian species, including wild cats. However, successes in application of assisted reproduction techniques in nondomestic felids have been sparse. Our objectives were to assess the effectiveness of alternating gonadotropin regimens on ovarian responses. Five adult female ocelots and four adult female tigrinus were treated four to six times, using alternating eCG/hCG and pFSH/pLH at 4-month intervals. Laparoscopies were done to assess follicular development and to collect oocytes from matures follicles. The average number of follicles and corpus luteum (CL) per stimulation was higher in ocelots (7.0 +/- 0.8; mean +/- S.E.M.) than in tigrinus (2.5 +/- 0.4; P < 0.05), but the percentage of mature oocytes did not differ between the two species (mean range, 54-55%). Within species, both gonadotropin regimens were equally effective in inducing follicular growth and oocyte maturation. The total number of ovarian structures and oocyte maturation percentages did not decrease in either species with sequential stimulations. In summary, female ocelots and tigrinus continued to respond to repeated alternating ovarian stimulation protocols. In conclusion, the use of alternating gonadotropin regimens may permit more intensive reproductive management in these endangered cats. PMID- 16472854 TI - The roles of progestagen and uterine irritant in the maintenance of cystic endometrial hyperplasia in the canine uterus. AB - Cystic endometrial hyperplasia (CEH) was induced in the left uterine horns of 14 mature ovariectomised greyhound bitches with an intra-luminal silk suture (uterine irritant) and treatment with estradiol benzoate and megestrol acetate (to simulate stages of a normal canine estrous cycle). Right uterine horns served as suture-free controls. From Day 30 of simulated diestrus, bitches received treatments of suture removal (n = 4), progestagen withdrawal (n = 5) or both (n = 5). Necropsies were performed 3 or 9 weeks later. At 3 weeks, severe cystic endometrial hyperplasia was present in all (6/6) left horns and in no (0/6) right horns. At 9 weeks, the left horns in 5/6 of bitches subjected to progestagen withdrawal had recovered (in varying degrees) from cystic endometrial hyperplasia, whereas no recovery was evident in the left horns of bitches (n = 2) that continued to receive progestagen. This study demonstrated that: (i) cystic endometrial hyperplasia was reversible upon withdrawal of progestagen; (ii) progestagen maintained cystic endometrial hyperplasia in the presence or absence of irritant; and (iii) persistent endometrial irritation in the absence of progestagen may not maintain cystic endometrial hyperplasia. PMID- 16472856 TI - Measuring nigrescence attitudes in school-aged adolescents. AB - In this study, we examined the reliability and validity of Cross Racial Identity Scale (CRIS; Vandiver, B. J., Cross Jr., W. E., Fhagen-Smith, P. E., Worrell, F. C., Swim, J. K., & Caldwell, L. D. (2000). The Cross Racial Identity Scale. Unpublished scale; Worrell, F. C., Vandiver, B. J., & Cross Jr., W. E., (2004). The Cross Racial Identity Scale: Technical manual (2nd ed.). Berkeley, CA: Author) scores in 143 middle and high school students. Exploratory factor analyses provided support for the six-factor structure of the CRIS, reliability estimates for the scores were in the moderate to high range, and subscale intercorrelations were low. Multicultural Inclusive scores were significantly correlated with age with high school students reporting significantly higher multicultural attitudes than middle school students. The authors concluded that the CRIS is appropriate for use with adolescent populations. PMID- 16472855 TI - Effect of in-office tooth bleaching on the microhardness of six dental esthetic restorative materials. AB - OBJECTIVES: The aim of this in vitro study was to evaluate the effect of the in office bleaching technique on the microhardness of six dental esthetic restorative materials. METHODS: Four composite resins (a hybrid, a flowable, a micro-hybrid and a nano-hybrid), an ormocer and a ceramic were tested, after the use of an in-office bleaching product. Fourteen specimens of each composite and the ormocer were fabricated and randomly divided into two groups of seven samples each. One group was polished and the other group remained unpolished. For the ceramic, seven polished samples were fabricated. Two samples of each group were used as negative controls. The specimens were bleached for 15, 30 and 45min. Five Knoop microhardness measurements were made on each sample, for each of the following periods tested: before bleaching, after 15, 30 and 45min of bleaching, 24h and 1 month after the bleaching procedure. Data were analyzed by the repeated measures analysis of variance with three between factors and one within. RESULTS: The differences in the microhardness values between the bleached and the control samples for the composites and the ceramic, were not statistically significant (hybrid: p=0.264; flow: p=0.584; micro-hybrid: p=0.278; nano-hybrid: p=0.405; ceramic: p=0.819). For the ormocer, although bleaching did not have any significant effect on the unpolished samples (p=0.115), it caused an increase on microhardness of the polished samples. SIGNIFICANCE: Bleaching with 38% hydrogen peroxide does not reduce the microhardness of the restorative materials tested. Therefore, no replacement of restorations is required after bleaching. PMID- 16472857 TI - Gain of 1q in pediatric myelodysplastic syndromes. AB - The presence of acquired clonal cytogenetic abnormalities in hematopoietic cells is one of the diagnostic hallmarks of myelodysplastic syndromes (MDS). Such anomalies may help in defining prognostic groups. We analyzed eight pediatric MDS, and herein describe three new cases, one de novo and two therapy-related, presenting an unbalanced rearrangement of 1q: one of them resulted in a derivative chromosome 6 apparently identical to a previously described one. We also review all the cases of gain of 1q reported in de novo and therapy-related childhood MDS. PMID- 16472858 TI - Evaluation of a novel short polyadenylation signal as an alternative to the SV40 polyadenylation signal. AB - The soluble neuropilin-1 (sNRP-1) gene employs an extremely short dual function polyadenylation (pA) signal/stop codon that is efficient for termination of gene transcription and translation in vivo. However, the functionality and usefulness of this signal in regard to other genes is unknown. This quantitative study compares the levels of humanized Renilla green fluorescent protein (hrGFP) mRNA polyadenylated with either the sNRP-1 pA signal or the much larger, more widely used SV40 pA signal. We show that the overall starting copy number of hrGFP for equally loaded RNAs is equivalent between the two groups. Our data show little to no difference between levels of mRNA generated by the sNRP-1 polyadenylation signal and the SV40 polyadenylation signal despite a remarkable size difference in signal length. An extremely short polyadenylation signal could potentially alleviate gene insert size restrictions associated with cloning or with therapeutic vectors such as adeno-associated virus. PMID- 16472859 TI - Molecular classification of IncP-9 naphthalene degradation plasmids. AB - A large collection of naphthalene-degrading fluorescent Pseudomonas strains isolated from sites contaminated with coal tar and crude oil was screened for the presence of IncP-9 plasmids. Seventeen strains were found to carry naphthalene catabolic plasmids ranging in size from 83 to 120 kb and were selected for further study. Results of molecular genotyping revealed that 15 strains were closely related to P. putida, one to P. fluorescens, and one to P. aeruginosa. All catabolic plasmids found in these strains, with the exception of pBS216, pSN11, and p8909N-1, turned out to belong to IncP-9 beta-subgroup. Plasmids pBS216, pSN11, and p8909N-1 were identified as members of IncP-9 delta-subgroup. One plasmid, pBS2, contains fused replicons of IncP-9beta and IncP-7 groups. RFLP analyses of the naphthalene catabolic plasmids revealed that organisation of the replicon correlates well with the overall plasmid structure. Comparative PCR studies with conserved oligonucleotide primers indicated that genes for key enzymes of naphthalene catabolism are highly conserved among all studied plasmids. Three bacterial strains, P. putida BS202, P. putida BS3701, and P. putida BS3790, were found to have two different salicylate hydroxylase genes one of which has no similarity to the "classic" enzyme encoded by nahG gene. Discovery of a large group of plasmid with unique nahR suggested that the regulatory loop may also represent a variable part of the pathway for catabolism of naphthalene in fluorescent Pseudomonas spp. PMID- 16472860 TI - Fine specificity and sequence of antibodies directed against the ectodomain of matrix protein 2 of influenza A virus. AB - The ectodomain of matrix protein 2 (M2e) has remained remarkably conserved amongst human influenza A viruses and is a target for Abs with protective activity. For these reasons, M2e is being investigated for its potential as a broadly protective influenza A virus vaccine. Here, we report on the fine specificity and sequence of seven M2e-specific mAbs isolated from three BALB/c mice after different immunization protocols. The mAbs recognized epitopes comprised within a 13aa long peptide corresponding to M2e(4-16). They originated from 4 distinct precursor B cells and showed a highly restricted variable (V) gene usage, in that their heavy chain V regions were all formed by the same V(H), D and J(H) gene segments and their light chain V regions made use of only two distinct Vkappa genes (Vkappa19-15/IGKV6-15 and Vkappa8-30/IGKV8-30; NCBI/IMGT annotation, respectively). The consensus sequence of the expressed V(H) genes belongs to the J558/HV1 family. It showed 96% identity with the BALB/c germline gene J558.n/IGHV1S137 and 100% identity with a V(H) gene expressed by several BALB/c B-1 B cells. This suggests that the consensus sequence is that of a functional BALB/c germline V(H) gene. The genetic restriction of this response may in part underlie the generally poor M2e-specific Ab response induced by infection. PMID- 16472861 TI - Compstatin inhibits complement activation by binding to the beta-chain of complement factor 3. AB - Compstatin is a peptidic complement inhibitor that prevents the cleavage of complement factor 3 (C3) by C3 convertase. Compstatin differs from other C3 regulatory proteins, such as complement receptor (CR) 1 and decay-accelerating factor (DAF), in that it binds native as well as activated C3 fragments and acts through mechanisms that do not involve the destabilization of the C3 convertase or the accelerated degradation of C3b. Compstatin's activity most likely relies on its affinity for native C3 and the conformational change that results upon binding with C3. Although the intermolecular interactions between compstatin and C3 have been studied, the identity of the targeted region on C3 is still elusive. To address this issue, we synthesized a photo-crosslinking compstatin analog and used it to probe C3 for sites of interaction. We identified a 40-kDa region at the C-terminus of the beta-chain of C3 that included the binding site of the compstatin analog. The specificity of the binding was confirmed by inhibition studies, which showed reduced crosslinking signal after pre-incubation of C3 with compstatin but not with various inactive analogs. Binding studies performed with a recombinant homolog of the 40-kDa region confirmed these findings. Five smaller recombinant proteins corresponding to various overlapping regions of the 40-kDa fragment did not bind compstatin, suggesting that a proper protein conformation, only found in larger fragments, is required for compstatin binding. The identified region on the beta-chain has, thus far, not been implicated in C3 cleavage or interactions with other proteins. Therefore, further research on this part of the C3 molecule may have implications for studies on the regulation of C3 cleavage, as well as for complement-based drug design. PMID- 16472862 TI - Gold(III) compounds of 1,4,7-triazacyclononane showing high cytotoxicity against A-549 and HCT-116 tumor cell lines. AB - Two gold(III) compounds [Au(TACN)Cl(2)]Cl (1) and [Au(TACN)Cl(2)][AuCl(4)] (2) (where TACN=1,4,7-triazacyclononane), have been synthesized and characterized by electrospray ionization mass spectrometry (ESI-MS), (1)H NMR spectroscopy and elemental analyses. The structure of compound 2 was determined by X-ray crystallography, in which TACN coordinates to the gold(III) center in a bidentate mode and the unbound amine group forms a very short intramolecular Au-H(-N) contact (1.91A). Biological activity data showed that compound 1 is more cytotoxic than cisplatin against A-549 and HCT-116 tumor cell lines. The interactions of compound 1 with CT-DNA were studied by UV-Vis, fluorescence and CD spectroscopy, which suggests that compound 1 can induce the distortion of DNA double helix. PMID- 16472863 TI - Copper-containing nitrite reductase: a DFT study of nitrite and nitric oxide adducts. AB - Copper-containing nitrite reductases (Cu-NIRs) reduce nitrite to NO. Reported here are DFT (density functional theory) results on models of the Cu-NIR active site bound to nitrite and nitric oxide. The Cu-NIR active site appears to have been designed to exclude N-nitrite binding even though N-O bond cleavage would be equally facile in the N- and O-isomers. The active site also appears to force a side-on coordination of the end-product, nitric oxide. The latter feature has to rely on the sterics of the active site to destabilize, thermodynamically speaking, the Cu-NO adduct; under these conditions, the absence of N-nitrite coordination is proposed to be merely a side-effect. For the Cu(II)-NO adduct, sterical crowding appears to also favour the Cu-NO electromer over Cu(I)-NO+, helping to avoid the potentially damaging chemistry associated with an NO+ moiety. These conclusions are in reasonable agreement with previous conclusions drawn from experiment [Science 304 (2004) 867]. PMID- 16472864 TI - Potassium channel blockers in multiple sclerosis: neuronal Kv channels and effects of symptomatic treatment. AB - Multiple sclerosis (MS) is an inflammatory disease of the central nervous system (CNS) characterized by demyelination, with a relative sparing of axons. In MS patients, many neurologic signs and symptoms have been attributed to the underlying conduction deficits. The idea that neurologic function might be improved if conduction could be restored in CNS demyelinated axons led to the testing of potassium (K(+)) channel blockers as a symptomatic treatment. To date, only 2 broad-spectrum K(+) channel blockers, 4-aminopyridine (4-AP) and 3,4 diaminopyridine (3,4-DAP), have been tested in MS patients. Although both 4-AP and 3,4-DAP produce clear neurologic benefits, their use has been limited by toxicity. Here we review the current status of basic science and clinical research related to the therapeutic targeting of voltage-gated K(+) channels (K(v)) in MS. By bringing together 3 distinct but interrelated disciplines, we aim to provide perspective on a vast body of work highlighting the lengthy and ongoing process entailed in translating fundamental K(v) channel knowledge into new clinical treatments for patients with MS and other demyelinating diseases. Covered are (1) K(v) channel nomenclature, structure, function, and pharmacology; (2) classic and current experimental morphology and neurophysiology studies of demyelination and conduction deficits; and (3) a comprehensive overview of clinical trials utilizing 4-AP and 3,4-DAP in MS patients. PMID- 16472865 TI - Determination of amino acids in saliva using capillary electrophoresis with fluorimetric detection. AB - In the present study a sensitive method for the quantification of main free amino acids in saliva using capillary electrophoresis with laser induced fluorescence detection was developed. As background electrolyte 20 mM borate buffer pH 9.5 was used. Amino acids were separated after derivatization with fluorescein isothiocyanate (FITC) and the conditions for derivatization were optimized. The main amino acids occurring in saliva (Pro, Ser, Gly and Glu) were separated in less than 7 min. The parameters of validation such as linearity of response, precision and detection limits were determined. The detection limits were obtained in the range from 0.1 to 2.4 nM. The developed method was employed for determination of amino acids in real saliva samples. PMID- 16472866 TI - Low-cost conversion of the Polaroid MD-4 land camera to a digital gel documentation system. AB - A simple, inexpensive design is presented for the rapid conversion of the popular MD-4 Polaroid land camera to a high quality digital gel documentation system. Images of ethidium bromide stained DNA gels captured using the digital system were compared to images captured on Polaroid instant film. Resolution and sensitivity were enhanced using the digital system. In addition to the low cost and superior image quality of the digital system, there is also the added convenience of real-time image viewing through the swivel LCD of the digital camera, wide flexibility of gel sizes, accurate automatic focusing, variable image resolution, and consistent ease of use and quality. Images can be directly imported to a computer by using the USB port on the digital camera, further enhancing the potential of the digital system for documentation, analysis, and archiving. The system is appropriate for use as a start-up gel documentation system and for routine gel analysis. PMID- 16472867 TI - Filtration disrupts synaptosomes during radiochemical analysis of serotonin uptake: comparison with chronoamperometry in SERT knockout mice. AB - Radiochemical methods have failed to reveal decreases in synaptosomal serotonin uptake in mice lacking one functional copy of the serotonin transporter (SERT) gene. By contrast, uptake rates determined by chronoamperometry in synaptosomes from SERT+/- mice show gene-related reductions. We revisited [(3)H]5-HT uptake in SERT knockout mice to determine the effects of inclusion of O(2) in the incubation buffer on the kinetic parameters obtained by this method. In oxygenated synaptosomes prepared from frontal cortex and striatum, modest 25 and 35% reductions in radiolabeled 5-HT uptake were detected in SERT+/- versus SERT+/+ mice. However, even in the presence of O(2), no differences in [(3)H]5-HT uptake were detected between SERT+/- and SERT+/+ mice in brain stem in contrast to 60% reductions determined by chronoamperometry. Moreover, while inclusion of O(2) modestly increased the rates of [(3)H]5-HT uptake, rates determined by chronoamperometry in the presence of O(2) were 40-fold greater than those determined radiochemically. We present evidence that the filtration process used in the radiochemical method leads to substantial loss of transported 5-HT resulting in lower apparent uptake rates. These findings explain the relative insensitivity of radiochemical methods for determining biologically important alterations in uptake such as those occurring between SERT+/- and SERT+/+ mice and in response to O(2). PMID- 16472868 TI - Tissue affinity of the infusate affects the distribution volume during convection enhanced delivery into rodent brains: implications for local drug delivery. AB - Convection-enhanced delivery (CED) is a recently developed technique for local delivery of agents to a large volume of tissue in the central nervous system (CNS). We have previously reported that this technique can be applied to CNS delivery of nanoparticles including viruses and liposomes. In this paper, we describe the impact of key physical and chemical properties of infused molecules on the extent of CED-mediated delivery. For simple infusates, CED distribution was significantly increased if the infusate was more hydrophilic or had less tissue affinity. Encapsulation of tissue-affinitive molecules by neutral liposomes significantly increased their tissue distribution. The poorer brain distribution observed with cationic liposomes, due to their greater tissue affinity, was completely overcome by PEGylation, which provides steric stabilization and reduced surface charge. Finally, liposomal encapsulation of doxorubicin reduced its tissue affinity and substantially increased its distribution within brain tumor tissue. Taken together, the physical and chemical properties of drugs, small molecules and macromolecular carriers determine the tissue affinity of the infusate and strongly affect the distribution of locally applied agents. Thus, an increased and more predictable tissue distribution can be achieved by reducing the tissue affinity of the infusate using appropriately engineered liposomes or other nanoparticles. PMID- 16472870 TI - Selective MCA occlusion: a precise embolic stroke model. AB - The present study describes a method for improving the precision and accuracy of clot placement within the middle cerebral artery (MCA) of rats, utilizing a micro catheter and laser Doppler flowmetry. This technique reduces the size of clot needed to achieve stable occlusion with no failed embolizations and a low percentage of early recanalizations. Infarctions were consistent in both size and distribution within the MCA perfusion territory. Selective embolization in aged animals (n = 10) resulted in substantially larger infarctions than those seen in aged animals (n = 10) following non-selective embolization (P < 0.05), or young animals (n = 10) subjected to filamentous occlusion (P < 0.001). Clots were localized to the MCA by direct examination at 0, 60 and 120 min post-embolization (n = 14). All aged animals surviving 24h exhibited moderate to severe functional deficits, with selectively occluded animals having a higher mean score on the modified neurologic severity scale (P = 0.002). This model provides a highly reproducible method for embolization of the MCA and reliable reperfusion with rt PA. PMID- 16472869 TI - Motor unit synchronous firing as revealed by determinism of surface myoelectric signal. AB - Information on motor strategies can be extracted from the surface electromyogram (EMG) by non-linear methods. The percentage of determinism (%DET) obtained from recurrence quantification analysis (RQA) may be a sensitive variable to detect synchronous motor unit behaviour. The purpose of the present study was to validate this methodology by comparing it with an established technique estimating the degree of synchronization of pairs of voluntary activated motor units from the correlation of their firing in the time-domain. Single motor unit activity was recorded in extensor carpi radialis (ECR) muscle by pairs of tungsten microelectrodes inserted into the muscle belly. Cross-correlation analysis was performed in order to determine synchronization peak area by computing synchronous impulse probability. Surface EMG activity was recorded in parallel by electrodes placed over the skin of the same muscle and %DET was used as a measure of synchronous activity. The %DET appeared to be a valid measure of synchronization yielding results comparable to those obtained with cross correlation analysis. Increases in %DET (t = 64.59, P < 0.0001) highly correlated (r2 = 0.70, P = 0.0013) with pharmacologically induced increases in the synchronization activity of pairs of ECR motor units (t = 8.71, P < 0.0001). RQA may be used as an alternative methodology for testing synchronous motor unit behaviour from surface EMG under physiological and pathological conditions. PMID- 16472872 TI - Immune escape of equine herpesvirus 1 and other herpesviruses of veterinary importance. AB - Equine herpesvirus (EHV)-1 is a pathogen of horses, well known for its ability to induce abortion and nervous system disorders. Clinical signs may occur despite the presence of a virus-specific immune response in the horse. The current review will summarize the research, on how, EHV-1-infected cells can hide from recognition by the immune system. Research findings on immune evasion of EHV-1 will be compared with those of other herpesviruses of veterinary importance. PMID- 16472871 TI - Detection of equine herpesvirus-specific effector and memory cytotoxic immunity in the equine upper respiratory tract. AB - Immunological protection of horses from equine herpesvirus-1 (EHV-1) infection and disease depends on the cooperation of virus-specific humoral and cellular immune responses. EHV-specific mucosal immunity may be an important component of such immune responses. This study demonstrates the induction of anti-EHV cytotoxic cellular immune responses in various mucosal and systemic lymphoid tissues associated with the upper respiratory tract (URT) of the horse. Four young horses (1-2 years of age) were inoculated intranasally with the Army 183 strain of EHV-1 and euthanized 1 week later. One untreated foal served as a non infected control. Mucosa-associated tonsillar tissues, draining lymph nodes and PBMC were harvested. Virus-specific memory and effector cytolytic activity were individually assessed using 4 h chromium release assays, with and without in vitro restimulation with EHV-1, respectively. EHV-specific cytotoxic activity was detected ex vivo in several URT-associated mucosal lymphoid tissues of horses, particularly within the lining of the nasopharynx, a principal site of EHV-1 replication. This activity was also detected in the circulation of some horses 1 week post-challenge. Virus-specific memory cytotoxic activity was elevated in the circulation, and detectable in the draining lymph nodes of all horses following challenge infection. PMID- 16472873 TI - Inhibition of the development of chronic experimental autoimmune encephalomyelitis by laquinimod (ABR-215062) in IFN-beta k.o. and wild type mice. AB - Laquinimod is a novel oral immunomodulatory substance, which is currently developed for the treatment of multiple sclerosis (MS). The ability of laquinimod to inhibit disease development was investigated in chronic experimental autoimmune encephalomyelitis (chEAE) in IFN-beta k.o. mice and wild type mice. Laquinimod was shown to inhibit both disease development and histopathological changes in the CNS. Furthermore, laquinimod was found to be independent of endogenous IFN-beta for its effect in chEAE. When laquinimod was combined with exogenous IFN-beta, a synergistic disease inhibitory effect was seen. These findings using laquinimod in preclinical disease models for MS emphasize the potential of laquinimod in the future treatment of MS also in patients that do not respond to IFN-beta monotherapy. Furthermore, the results indicate that laquinimod may favourably be combined with IFN-beta. PMID- 16472874 TI - Detection of three porcine vesicular viruses using multiplex real-time primer probe energy transfer. AB - Rapid identification of the etiologic agent in infected animals is important for the control of an outbreak of vesicular disease in livestock. We have in the present study developed a multiplex real-time reverse transcription-PCR, based on primer-probe energy transfer (PriProET), for simultaneous detection and differentiation of three Office International des Epizooties (OIE) classified vesicular viruses: foot-and-mouth disease virus, vesicular stomatitis virus and swine vesicular disease, causing clinically indistinguishable vesicular diseases in swine. The multiplex assay consists of extraction of total RNA from clinical samples; reverse transcription to cDNA using random primers and one-tube real time amplification of cDNA using multiplex PriProET with specific fluorescent labelled primers and probes for detection of the three viruses from the vesicular disease complex. The probes are labelled with unique reporter fluorophores, which during amplification are excited by donor fluorophores incorporated in the 5' end of specific amplicons by primer extension. The sensitivity of the multiplex assay was approximately 100 TCID(50), which is 10-fold lower compared to the individual PriProET assays for the three vesicular viruses. PMID- 16472875 TI - Real-time PCR protocol for the detection of porcine parvovirus in field samples. AB - This report describes a real-time polymerase chain reaction assay with SYBR Green for detection of a broad range of porcine parvoviruses (PPV) and accurate virus quantification in porcine tissues. The assay targets the VP2 gene of PPV and the porcine genomic c-myc gene for normalization. The detection limit of the SYBR Green reaction was shown to be equivalent to 6 x 10(0) to 6 x 10(1) PPV copies/reaction and the overall detection limit equivalent to 0.1 TCID(50)/100 microl. The assay was linear over a 10(7) dilution range of template concentrations. Other porcine pathogens involved in reproductive disorders such as porcine circovirus 2 (PCV-2), porcine reproductive and respiratory virus (PRRSV), Aujeszky's disease virus (PRV) and other parvoviruses such as feline parvovirus (FPV), canine parvovirus (CPV), minute virus of canines (MVC) and a human parvovirus (B19) were not detected by this assay. PMID- 16472877 TI - In vitro selection of mutations in human immunodeficiency virus type 1 reverse transcriptase that confer resistance to capravirine, a novel nonnucleoside reverse transcriptase inhibitor. AB - Capravirine (CPV; formerly AG1549 and S-1153) is a novel, nonnucleoside reverse transcriptase inhibitor (NNRTI) of human immunodeficiency virus type 1 (HIV-1) that has demonstrated potent in vitro antiviral activity against several HIV-1 laboratory strains and clinical isolates with EC50 values ranging from 0.7 to 10.3 nM. In this study, we evaluated the resistance and cross-resistance profiles of CPV through selection of resistant HIV-1 variants from in vitro serial passage of HIV-1 NL4-3 and HIV-1 IIIB and by performing susceptibility assays on HIV-1 variants constructed to contain CPV-specific amino acid substitutions in reverse transcriptase (RT). Results demonstrate that HIV-1 variants selected at increasing CPV concentrations contained multiple substitutions in diverse patterns including L100I, Y181C, G190E and/or L234I in various combinations with K101R/E, K103T, V106A/I, V108I, E138K, T139K, A158T, V179D/I/G, Y188D, V189I, G190A, F227C, W229R, L234F, M230I/L and P236H/T. Interestingly, HIV-1 variants constructed to contain the T215Y zidovudine (AZT)-resistance associated substitution with CPV-resistance associated substitutions V106A, Y181C, F227C, F227L, L234I or V106A/F227L demonstrated 2.4-5.4-fold increased susceptibility to CPV. Results also demonstrate that the CPV-resistance associated substitutions Y181C, F227C, F227L and L234I reverse the phenotypic resistance to AZT conferred by the T215Y substitution. PMID- 16472876 TI - Mapping brain maturation. AB - Human brain maturation is a complex, lifelong process that can now be examined in detail using neuroimaging techniques. Ongoing projects scan subjects longitudinally with structural magnetic resonance imaging (MRI), enabling the time-course and anatomical sequence of development to be reconstructed. Here, we review recent progress on imaging studies of development. We focus on cortical and subcortical changes observed in healthy children, and contrast them with abnormal developmental changes in early-onset schizophrenia, fetal alcohol syndrome, attention-deficit-hyperactivity disorder (ADHD) and Williams syndrome. We relate these structural changes to the cellular processes that underlie them, and to cognitive and behavioral changes occurring throughout childhood and adolescence. PMID- 16472879 TI - Repeated administration of dopaminergic agents in the nucleus accumbens and morphine-induced place preference. AB - In the present study, the effects of repeated intra nucleus accumbens (intra-NAc) injections of dopamine receptor agents on morphine-induced conditioned place preference (CPP) in rats were investigated by using an unbiased 3-days schedule of place conditioning design. The animals receiving once daily subcutaneous (s.c.) injections of morphine (0.5-7.5mg/kg) or saline (1.0 ml/kg, s.c.) showed a significant place preference in a dose-dependent manner. The maximum response was observed with 5mg/kg of the opioid. Three days intra-NAc injections of apomorphine (0.5 and 1 microg/rat) followed by 5 days free of the drug, increased or decreased, respectively CPP induced by the lower dose of morphine (0.5mg/kg, s.c.). Morphine-induced CPP was also significantly increased in the animals that had previously received the 3-days intra-NAc injections of SKF 38393 (4 and 8 microg/rat) or quinpirole (2 and 4 microg/rat, intra-NAc). The CPP induced by a higher dose of morphine (5mg/kg, s.c.) was significantly decreased in the animals that had previously received the 3-days SCH 23390 (0.005 and 0.01 microg/rat; intra-NAc). On the other hand, the CPP induced by morphine (5mg/kg, s.c.) was significantly increased in the animals that had previously received the 3-days sulpiride administration (5 microg/rat, intra-NAc). The 3-days administration of apomorphine, SKF 38393 or quinpirole, but not SCH 23390 and sulpiride reduced the locomotor activity in the test session. It is concluded that repeated injections of dopamine receptors agents followed by 5 days free of the drugs in the NAc can affect morphine reward. PMID- 16472878 TI - Zaprinast consolidates long-term memory when administered to neonate chicks trained using a weakly reinforced single trial passive avoidance task. AB - A weakly reinforced variant of the single trial passive avoidance task developed for the day-old chick typically fails to consolidate long-term memory. However, administration of zaprinast, a phosphodiesterase (PDE) type 5 inhibitor, (ic; 10 microl/side) immediately post-training resulted in a dose-dependent increase in retention at test 180 min post-training. Further, 100 microM zaprinast resulted in high levels of retention at test 180 min post-training when administered from 10 min before training to 10 min after training. Finally, 100 microM zaprinast, when administered immediately post-training, resulted in the consolidation of long-term memory at a number of times of test extending as late as 24 h post training. Inhibition of PDE type 5 is known to increase cellular cGMP levels. Previous investigations using a strongly reinforced variant of this task have suggested a role for cGMP in memory retrieval, we now postulate that cGMP is also necessary for memory formation in chicks trained using passive avoidance. PMID- 16472880 TI - Short-term potentiation at the parallel fiber-Purkinje cell synapse. AB - Changes in synaptic efficacy at the parallel fiber (PF)-Purkinje cell (PC) synapse are postulated to be a cellular basis for motor learning. Although long term efficacy changes lasting more than an hour at this synapse, i.e., long-term potentiation and depression, have been extensively studied, relatively short lasting synaptic efficacy changes, namely short-term potentiation (STP) lasting for tens of minutes, have not been discussed to date. Here we report that this synapse shows an apparent STP reliably by a periodic burst pattern of homosynaptic stimulation. This STP is presynaptically expressed, since it accompanies with a reduced paired-pulse facilitation and is resistant to postsynaptic Ca(2+) reduction by BAPTA injection or in P/Q-type Ca channel knockout cerebella. This novel type of synaptic plasticity at the PF-PC synapse would be a clue for understanding the presynaptic mechanisms of plasticity at this synapse. PMID- 16472881 TI - Use of a serological approach for prediction of Salmonella status in an integrated pig production system. AB - Relevance of a Salmonella serological detection technique was studied from complete results obtained from 9 pigs fattening units. Feces and overshoes were sampled at different periods after starting fattening (2, 3 and 4 months) while caecal contents were taken on the slaughter line. The bacteriological technique used was based on a Diasalm enrichment and a commercial test was used for serology on an average of ten animals per batch. The aim of this work was to establish a correlation between serological results obtained at slaughter (10 samples/batch) and bacteriological results. In this context, two types of logistic regression models were tested by considering alternatively serology and Salmonella detection in caecal contents as the dependent variables. Firstly, beside the fact that all logistic regression models show weak correlations, the first finding was that positive results in overshoes taken at 2 and 3 months are slightly correlated with serological status of herds (odds-ratios of 4.96 and 2.55). Secondly, when batches were characterized as positive on the basis of serological results, the probability of Salmonella recovery in caecal contents was higher than when the batches were considered as negative (odds-ratios comprised between 4.36 and 5.81). A major conclusion is that serology can be used to follow the improvement of an integrated pig production system, but is not the unique solution for assessing risk of Salmonella shedding from specific herds. PMID- 16472882 TI - Hepatitis E virus genotyping based on full-length genome and partial genomic regions. AB - Some genomic regions for hepatitis E virus (HEV) genotyping have been reported to correlate well with the results from the phylogenetic analyses on the basis of the complete genome. However, few studies have systemically investigated the genomic regions for HEV genotyping using a combined phylogenetic and statistical approach. A consensus region for HEV genotyping has not been determined. In this study the nucleotide identities and genetic distances of 24 partial genomic regions and the complete genome sequences of 37 HEV strains were compared statistically. It was demonstrated with both one-way ANOVA and two-way ANOVA that only one genomic region in RNA-dependent RNA polymerase domain (4254-4560nt) for which there were no significant differences when compared with the full-length genome (P>0.05). The same four genotypes were identified by phylogenetic analysis based on this statistically predicted region identified as for the complete genome. RT-PCR amplification of HEV strains from all four genotypes confirmed conservation of the flanking primer sites of this region. Serum samples from 20 patients with a clinical diagnosis of hepatitis E were further analyzed by PCR using the same primers, 13 were positive and could be classified into genotype 4. These data strongly suggested that this newly identified region could be used for future HEV genotyping. PMID- 16472883 TI - [Disabled workers with motor impairments: data from an occupational health service]. AB - INTRODUCTION: Mediclen is an occupational health service in charge of following up 36,736 workers (divided among 1770 companies) in 3 cities of an area near Paris. The employment rate of disabled people among the French population is not well known (rough estimate 4.4%), and few studies have reported on the situation of workers with a motor impairment. SUBJECTS AND METHOD: The recent computerization of medical records allowed us to identify 195 workers considered disabled by the French administration (i.e. 0.55% of the 36,736 workers followed up in 2002). Among these, 26 had a motor impairment. RESULTS: Twenty-one neurological disabilities were central and 5 were peripheral or neuromuscular. The workers were 44-years-old. Only two workers had a severe handicap. Companies had to adapt workstations for half of the workers, with the advice of neurologists (7 of 10 advice given) and once a physical medicine doctor. DISCUSSION-CONCLUSION: The integration of people with motor impairments into the world of work is rare and difficult. This practical experience showed the difficulties people with motor impairment face. Close collaboration of physical medicine services with occupational health services is necessary to improve the integration of this population into the world of work. PMID- 16472884 TI - [Multiresistant bacteria colonisation in rehabilitation units]. AB - The objective of this study was to evaluate the management of multiresistant bacteria infection in French rehabilitation units. We e-mailed a questionnaire to French rehabilitation practitioners asking them what they did and what they knew about multiresistant bacteria colonisation. Their answers are described and analysed. We received 60 answers representing most of the country's rehabilitation centers. Routine screening was rare, but multiresistant bacteria were often looked for when certain risk factors were present. Methicillin resistant Staphylococcus aureus and multiresistant enterobacteria were the usual target germs. In general, respondents asked for evidence-based recommendations that are also applicable without too great a prejudice (for the rehabilitation as well as for the psychological state) for a patient hospitalised in a rehabilitation unit. The legal risks and the costs involved are also to be tested. Many questions lay unanswered, although the literature is rich in descriptions of this area. The opposition between the medical advice of isolation and the readaptation objectives of a rehabilitation unit are strong. This study aims to highlight the most important uncertainties so as to promote prospective studies and enlighten the legal authorities. PMID- 16472885 TI - Allocation of labour to somatic and psychiatric specialist care--the effects of earmarked grants. AB - Until 2002, counties were responsible for providing both somatic and psychiatric specialized health care services in Norway. The financing arrangement of the counties consisted of fixed local taxes, a general block grant and different types of earmarked grants from the national government. Since 1997, earmarked conditional grants related to DRG-activity have been used for somatic services, whereas earmarked unconditional grants have been used for mental health care services from approximately the same time. This paper analyse the price and revenue effects of grants on the allocation of labour, with special attention to the two types of earmarked grants: conditional and unconditional. Theoretically, labour (as an index of production output) is assumed to be allocated to somatic and psychiatric services dependent upon revenues (taxes, block grants and earmarked unconditional grants), price per labour-year (that among other things are affected by earmarked conditional grants) and the preferences of the local government (which relate to the age structure and population density of the county). We assume that a conditional grant reduces the net price of labour and thereby increases the (relative) demand, whereas an unconditional grant adds to the other revenues of the county and do not affect the relative allocation of labour. Data from a panel of 18 counties for the period 1992-2001 is analysed using OLS with fixed effects. The results show revenue effects and direct price effects as expected. However, the assumption that unconditional grants do not affect relative allocation of labours is not supported. We find a positive effect of the unconditional grant to psychiatric care on the demand for labour in this sector. We interpret this as an effect of hierarchical governance such as supervision and monitoring that were remedies that were used together with the unconditional grant. PMID- 16472886 TI - Learning to cross boundaries: the integration of a health network to deliver seamless care. AB - We analysed the development of an integrated network from a learning perspective to see how care givers from different organisations were able to cross the professional and organisational boundaries that existed between them to make sure patients receive the right care, at the right moment, in the right place. We show how through a process of collective learning social contacts between health professionals increased and improved. These professionals learned to speak each other's language, learned how other professionals and organisations work and learned to look at the care process from a network perspective instead of only from a professional or organisational perspective. Through this learning process, they also experienced the limitations of standardizing knowledge in criteria, protocols and rules, and the value of direct contact for sharing information and knowledge, to ensure continuity in care. PMID- 16472887 TI - Health system factors impacting on delivery of mental health services in Russia: multi-methods study. AB - OBJECTIVE: To evaluate how the regulatory environment and health system organisation, financing and provider payment systems influence the delivery of mental health services in the Sverdlovsk region of the Russian Federation. METHODS: A multi-methods study was conducted including analysis of routine data and key informant interviews supplemented by analysis of published literature, legal and regulatory documents, ministerial orders and reports. RESULTS: Mental health care services are still largely provided in hospitals, although the need for more community-based and rehabilitation services is widely recognised. Resource allocation and provider payment systems remain largely unchanged from Soviet times and favour large inpatient institutions, creating incentives for hospitals to maintain a large number of beds and staff. Community-based social services and human resources remain limited, especially in the areas of social work, housing support and vocational rehabilitation, but co-operation across sectors at local level is growing. CONCLUSION: In Russia, the pace of reform in the mental health system will be helped if financial resource allocation mechanisms and provider payment systems are also reformed, so that resources follow individuals regardless of where they are treated. Such major health system shifts can only be achieved through changes at the Federal level and require major political will. Additional transitional funding is also required to help develop the necessary alternative community-based services. The nature of mental health disorders mean that this is not a problem faced within the health system alone, greater attention needs to be placed also on how to maximise the cross sector benefits especially with the social protection and employment sectors. PMID- 16472888 TI - A comparison of new drug availability in Canada and the United States and potential therapeutic implications of differences. AB - BACKGROUND: Claims are made that new valuable drugs are not available in Canada at the time that they are marketed in the United States. This study uses a convenience sample of new drugs marketed in the United States and determines how many of these products are initially unavailable in Canada and their therapeutic value. METHODS: Issues of the Canadian edition of The Medical Letter from May 12, 2003 to June 21, 2004 were hand searched for evaluations of new drugs and the following information was recorded: indication, availability in Canada and conclusions about therapeutic value. For drugs not available in Canada two clinical pharmacologists rated the therapeutic value of the products and the type of FDA review (standard or priority) was recorded. A database from the Therapeutic Products Directorate was searched to see if any of the drugs initially unavailable were subsequently marketed. RESULTS: Thirty-two of 37 drugs were not available in Canada. Between 9 and 11 of these products were rated as offering moderate to significant therapeutic gains. Twelve of the 32 drugs eventually were marketed in Canada. INTERPRETATION: Although the majority of new drugs marketed in the United States but not available in Canada do not offer any therapeutic advantage, between about a quarter and a third of may offer moderate to significant therapeutic gains. The reasons why these drugs are unavailable and how much their absence affects the treatment Canadians receive should be the subject of future research. PMID- 16472889 TI - The difference in mRNA expressions of hypothalamic CCK and CCK-A and -B receptors between responder and non-responder rats to high frequency electroacupuncture analgesia. AB - The present study was performed to determine whether the expression levels of the hypothalamic cholecystokinin (CCK) and its receptors are associated with the responsiveness to high frequency electroacupuncture (EA) analgesia in rats. EA stimulation (100 Hz, 0.5 ms pulse width, 0.2-0.3 mA) was delivered to the Zusanli (ST36) acupoint of male Sprague-Dawley rats for 20 min without anesthetics or holder restraint. The analgesic effect of EA was quantified using a tail flick latency test, and subsequently animals were allocated to responder or non responder groups. The hypothalamus of rats in each group was dissected and RNA was purified. The mRNA expressions of CCK, and CCK-A and -B receptor were determined by real-time RT-PCR. CCK mRNA levels were not significantly different in the two groups, whereas both CCK-A and -B receptors were significantly more expressed in non-responders. These results suggest that the level of CCK receptor mRNA expression in the hypothalamus, rather than CCK mRNA, has an important relationship with the individual variations to high frequency EA analgesia in rats. PMID- 16472890 TI - Molecular cloning of BNP from heart and its immunohistochemical localization in the hypothalamus of monkey. AB - Previous physiological studies have suggested central roles of brain natriuretic peptide (BNP). However, little information is available about the localization of BNP in the brain. In this study, we determined cDNA sequence encoding the entire coding region of prepro-BNP of Japanese and cynomologus monkeys, and then examined the immunohistochemical localization of BNP in the monkey hypothalamus. Japanese and cynomologus monkey prepro-BNP consisted of 132 amino acid residues with biologically active C-terminal 32 amino acids. Comparisons of deduced amino acid sequences among different species revealed high homology between monkey and human (91% in prerpro-BNP and 97% in the mature region). Immunohistochemical examination showed that BNP immunoreactive dots were observed in the paraventricular, periventricular, and supraoptic nuclei of the monkey hypothalamus. The present result suggests the central role of BNP in the neuroendocrine system in the hypothalamus. PMID- 16472891 TI - Synthesis of some new substituted triazolo [4,3-a][1,4] benzodiazepine derivatives as potent anticonvulsants. AB - Novel 8-chloro-6-(2-fluorophenyl)-1-(aryl)-4H-[1,2,4]triazolo[4,3 a][1,4]benzodiazepines (5a-f) were prepared by treating 7-chloro-5-(2 fluorophenyl)-1,3-dihydro-2H-1,4-benzodiazepine-2-thione with various aromatic acid hydrazides. The newly prepared compounds were characterized by spectral analysis. Compounds were tested for anticonvulsant activity. Four of the tested compounds such as 5a, 5d, 5e and 5f exhibited excellent anticonvulsant activity in comparison with standard drug, diazepam. PMID- 16472892 TI - Influence of the tunnel angle in ACL reconstructions on the biomechanics of the knee joint. AB - BACKGROUND: A high tension in anterior cruciate ligament grafts affects both graft and knee functional performance. Clinical observations suggest that impingement of the graft against the posterior cruciate ligament might cause high graft tensions. Also, meniscal injury has been well documented in association with damage in the anterior cruciate ligament. METHODS: In this paper, we present the results obtained in a three-dimensional finite element model of the human knee, corresponding to different aspects of anterior cruciate ligament reconstruction with bone-patellar tendon-bone grafts. This model was used to investigate the effect of the angle in the coronal plane of femoral and tibial tunnels. Firstly, graft tension was computed in a knee moved from 0 degrees to 60 degrees of flexion and the results were compared with experimental ones obtained by other authors. Secondly, the resulting kinematics under an anterior load of 134 N was compared to that of the intact knee. FINDINGS: The obtained results showed that the closest anterior tibial translation to that of the intact knee was obtained with femoral and tibial tunnels with angles of 60 degrees. In this same case, a lower graft tension was also obtained. The results demonstrated noticeable increases in the meniscal stresses after anterior cruciate ligament reconstruction. INTERPRETATION: Our results showed that impingement only depends on the femoral tunnel angle. On the contrary, laxity principally depends on the tibial tunnel angle. The angle of the femoral tunnel affects the graft tension while the tibial tunnel affects laxity, meniscal stresses and strains. PMID- 16472893 TI - Anemia in the elderly: current understanding and emerging concepts. AB - Anemia is currently defined by the World Health Organization (WHO) as a hemoglobin (Hb) level <13 g/dL in men and <12 g/dL in women. While estimates vary widely, nearly one quarter of community-based octagenerians and one half of the chronically ill elderly have Hb levels that satisfy a diagnosis of anemia according to these criteria. A growing body of evidence has linked adverse events with even "mild" anemia or low-normal Hb in the elderly. Recent studies suggest strongly that aging is associated with dysregulation of pro-inflammatory cytokines, most notably interleukin-6 (IL-6), which may negatively impact hematopoiesis, either by inhibition of erythropoietin (EPO) production or interaction with EPO receptors. Anemia in older individuals is associated with a very wide range of complications, including increased risk for mortality, cardiovascular disease, cognitive dysfunction, longer hospitalization for elective procedures and comorbid conditions, reduced bone density, and falls and fractures. Not surprisingly, anemia also has a significant effect on quality of life (QOL) in the elderly. Most anemia in older individuals results from iron deficiency, chronic inflammation, or chronic kidney disease, or it may be unexplained. Future research on anemia in the elderly should focus on the age related physiologic changes underlying this condition and whether anemia correction can reduce anemia-associated risks, and improve QOL. PMID- 16472895 TI - Is conditioning a useful framework for understanding the development and treatment of phobias? AB - Despite the prevalence of therapeutic interventions based on conditioning models of fear acquisition, conditioning has been seen by many as a poor explanation of how fears develop: partly because research on conditioning has become less mainstream and models of learning have become increasingly more complex. This article reviews some of what is now known about conditioning/associative learning and describes how these findings account for some early criticisms of conditioning models of fear acquisition. It also describes how pathways to fear such as vicarious learning and fear information can be conceptualised as forms of associative learning that obey the same learning rules. Some popular models of conditioning are then described with a view to highlighting the important components in learning. Finally, suggestions are made about how what we know about conditioning can be applied to improve therapeutic interventions and prevention programs for child anxiety. PMID- 16472894 TI - A remediation strategy based on active phytoremediation followed by natural attenuation in a soil contaminated by pyrite waste. AB - Phytoremediation of metal-polluted soils can be promoted by the proper use of soil amendments and agricultural practices. A 4-year phytoremediation programme was applied to a site affected by the toxic spill of pyrite residue at Aznalcollar (Spain) in 1998, contaminated with heavy metals (Zn, Cu, Pb, Cd) and arsenic. This consisted of active phytoremediation, using organic amendments (cow manure and compost) and lime and growing two successive crops of Brassica juncea (L.) Czern., followed by natural attenuation without further intervention. Changes in soil pH, extractable metal and As concentrations, organic carbon content and microbial biomass was evaluated. The initial oxidation of metal sulphides from pyrite residues released soluble metals and reduced soil pH to extremely acidic values (mean 4.1, range 2.0-7.0). The addition of lime (up to 64 t ha(-1)) increased soil pH to adequate values for plant growth, resulting in a significant decrease in DTPA-extractable metal concentrations in all plots. The natural attenuation phase showed also a decrease in extractable metals. Organic treatments increased the soil total organic carbon, which led to higher values of microbial biomass (11.6, 15.2 and 14.9 g kg(-1) TOC and 123, 170 and 275 microg g(-1) biomass-C in control, compost and manure plots, respectively). Active phytoremediation followed by natural attenuation, was effective for remediation of this pyrite-polluted soil. PMID- 16472896 TI - Anxiety of childhood and adolescence: Challenges and opportunities. AB - Despite significant advances in our understanding of anxiety in childhood and adolescence, the area is still in its infancy. However, this is an area that is attracting increasing interest from researchers and clinicians alike. This editorial describes some of the aspects of research and clinical attention that are likely to be most fruitful in the coming years, and focusses on some of the inter-related themes that have emerged from the six papers comprising this special edition of Clinical Psychology Review. The first theme concerns the quality and limited power of studies (particularly treatment trials) that have characterised this field. A number of the authors contributing to this edition have noted that this lack of investment in high quality, highly powered research has prevented many of the key questions from being answered. Second, there is growing awareness that we are under-investigating anxiety in younger children. Third, and relatedly, there is still a huge amount of work to be done in understanding the role of the family in child anxiety. What limited information that does exist is confusing and contradictory, and some suggestions for clarifications in this area are made. Finally, there is a plea for more developmentally appropriate, family-focussed and child-led models of anxiety in young populations. PMID- 16472897 TI - Non-epileptic seizures and child sexual abuse: a critical review of the literature. AB - Non-epileptic seizures have received a substantial amount of attention in the psychiatric and medical literature, but comparatively little attention from psychologists. Non-epileptic seizures resemble epileptic seizures but lack the physiological symptoms of genuine epilepsy and are psychological in origin. Many authors have emphasized the role that child sexual abuse may play in the etiology of this disorder. In the present paper, we provide a review of 34 studies examining this relationship, followed by a meta-analysis of 19 effect sizes. While our statistical results support the professed link between child sexual abuse and non-epileptic seizures, we suggest that because of research design limitations, it is premature to draw any definitive conclusions regarding a relationship. Eight of these research design limitations are identified and discussed (e.g., the absence of comparison groups; an explicit and public definition of child sexual abuse). Alternatives to a traditional psychoanalytic perspective that emphasizes the role of child sexual abuse in the etiology of NES are presented. Specific recommendations for future research are made and psychologists are strongly encouraged to play a more active role in both researching and treating non-epileptic seizures. PMID- 16472898 TI - Development of a reference dose for the persistent congeners of weathered toxaphene based on in vivo and in vitro effects related to tumor promotion. AB - Toxaphene is a mixture of chlorinated camphenes and bornanes that was produced and used in the United States until 1982. 1.3 million tons of toxaphene have been released worldwide. "Technical" toxaphene (TT) consists of a mixture of up to 800 different chemicals, known as congeners. TT weathers in the environment by both biotic and abiotic processes. The human body burden of toxaphene consists of only five persistent congeners that are not metabolized; three of these occur in considerably greater amounts than the other two. Because of the rapid metabolism and excretion of the non-persistent congeners, the persistent congeners that make up the human body burden most likely play a role in eliciting any potential adverse effects. EPA's toxicity assessment for TT is based on the occurrence of liver cancer in rodents, and considerable doubt exists whether this assessment is applicable to weathered toxaphene (WT). Using experimental results from European Union scientists, a reference dose (RfD) was developed for WT based on the three most persistent congeners that comprise the human body burden. The critical effect chosen was tumor promotion and this endpoint is considered protective for other endpoints as well. Although RfDs are typically derived for non-carcinogenic effects, the endpoint of tumor promotion is appropriate for RfD development because the experimental data suggest a dose threshold. The RfD for weathered toxaphene represented by the sum of the three major persistent congeners ( summation 3PC) is 2E-05 mg/kg-day. To apply this reference dose to a particular WT mixture, information is needed regarding the percentage of summation 3PC in the mixture. PMID- 16472899 TI - Exit, voice, governance and user-responsiveness: the case of English primary care trusts. AB - Hirschman contrasts exit and voice as 'recuperation' mechanisms for making organisations responsive to users. However, the emergence of health-care quasi markets and of network governance structures since Hirschman necessitate revising his theory, for they complicate the relationships between governance structures and recuperation mechanisms. Using a case study of nine primary care trusts (PCTs), this paper analyses the recuperation mechanisms, governance structures and relations between them in primary care in England. User voice can be exercised through dedicated networks besides hierarchies. As well as the 'user exit' described by Hirschman, two new 'exit' mechanisms now exist in quasi markets. Commissioner exit occurs when a third-party payer stops using a given provider. Professional proxy exit occurs when a general practitioner (GP) fund holder (or analogous budget-holder) behaves similarly. Neither exit mechanism requires the existence of mechanisms for user exit from healthcare purchasers, provided strong voice mechanisms exist instead to make commissioners responsive to users' demands. Establishing such voice mechanisms is not straightforward, however, as the experience of English PCTs illustrates. PMID- 16472900 TI - Marital status disparities in maternal smoking during pregnancy, breastfeeding and maternal depression. AB - One of the dramatic recent changes in family life in Western nations has been the rise in non-marital childbearing. Much of this increase is attributable to the growth in cohabitation. But in some countries, notably the UK (and the USA) this is much less the case with significant proportions of children being born to parents who are not living together. This study uses data from the Millennium Cohort Study, a British birth cohort established in 2001, to examine whether the closeness of the tie between parents, as assessed by their partnership status at birth, is related to smoking during pregnancy, breastfeeding and maternal depression. Four sets of parents are distinguished representing a hierarchy of bonding or connectedness: married and cohabiting parents, and two groups of solo mothers, those closely involved with the father at the time of the birth and those not in a relationship. Smoking in pregnancy, breastfeeding and maternal depression tests for trend, adjusted for socio-demographic factors, showed that there was a statistically increased risk of adverse health and health behaviours by degree of parental connectedness. There were also consistent and statistically significant differences between married and non-married mothers. Particularly noteworthy was the finding that cohabiting mothers have greater risk of adverse outcomes than married women. Among the non-married set, there were also differences in risk of adverse outcomes. For smoking in pregnancy, the key difference for continuing to smoke throughout the pregnancy lay between mothers involved with partners and those lacking an intimate relationship. For breastfeeding, stronger parental bonds were associated with initiation of breastfeeding, with a clear difference between cohabiting mothers compared to solo mothers. There was also an increased risk of maternal depression with looser parental bonding, and among non-married groups this increased risk was most noticeable among cohabiting mothers when compared with solo mothers. PMID- 16472902 TI - Use of reverse adipofascial flap in cutaneous defects of the distal third of the lower leg. AB - Lower limb reconstruction, especially for the aquilian and calcaneal regions, represent a great challenge for the plastic surgeon. Among the existing options of cutaneous coverage, the distally pedicled adipofascial flap may reach even the most distal zones. From April 1995 to May 2002, 15 adipofascial flaps to the cutaneous coverage of the distal-third leg were performed. The reconstruction was immediate in eight patients. In one case, partial tissue necrosis was observed. PMID- 16472903 TI - An innovative model for regulating supplement products: natural health products in Canada. AB - On 1 January 2004, Health Canada officially added a new term to the global list of synonyms for dietary supplements: natural health products (NHP). Developed with the intent of providing Canadian consumers with ready access to NHP that are safe, effective, and of high quality, the Natural Health Products Regulations (the NHP regulations) are applicable to the sale, manufacture, packaging, labelling, importation, distribution, and storage of NHP, and are administered by the recently formed Natural Health Products Directorate (NHPD) within Health Canada. This paper provides an overview of the process for regulating supplement products in Canada. PMID- 16472901 TI - Anterior medial temporal lobe activation during encoding of words: FMRI methods to optimize sensitivity. AB - The existence of a rostrocaudal gradient of medial temporal lobe (MTL) activation during memory encoding has historically received support from positron emission tomography studies, but less so from functional MRI (FMRI) studies. More recently, FMRI studies have demonstrated that characteristics of the stimuli can affect the location of activation seen in the MTL when those stimuli are encoded. The current study tested the hypothesis that MTL activation during memory encoding is related to the modality of stimulus presentation. Subjects encoded auditorily or visually presented words in an FMRI novelty paradigm. Imaging and analysis parameters were optimized to minimize susceptibility artifact in the anterior MTL. Greater activation was observed in the anterior than posterior MTL for both modalities of stimulus presentation. The results indicate that anterior MTL activation occurred during encoding, independent of stimulus modality and provide support for the hypothesis that verbal-semantic memory processing occurs in anterior MTL. The authors suggest that technical factors are critical for observing the rostrocaudal gradient in MTL memory activation. PMID- 16472904 TI - Influence of carbamide peroxide on enamel fluoride uptake. AB - OBJECTIVE: Aim of the study was to evaluate the influence of carbamide peroxide (CP) on enamel fluoride uptake by comparing enamel fluoride uptake from a 1% amine fluoride (AmF) gel with the fluoride acquisition from a 10% carbamide peroxide agent supplemented with 1% AmF. MATERIALS AND METHODS: Three enamel cylinders (4mm in diameter) were prepared from the buccal surfaces of 60 bovine incisors. One sample of each tooth was used for determination of baseline fluoride content of the respective tooth. The two remaining samples were allocated to the experimental series 1 or 2, respectively. Each series consisted of five experimental groups (A-E, n=12) and differed with respect to the length of the treatment period with the gels (A-D). The experimentally designed gels (pH 5.5) used in the study were as follows: A (10% CP), B (10% CP, 1% F(-) as AmF), C (1% F(-) as AmF), D (no CP, no F(-)) and were formulated on the same basis. The enamel samples were covered for 4h with the respective gel at 37 degrees C and were then transferred to artificial saliva for 20 h (series 1). The samples of group E served as controls and were not treated with a gel. In series 2, treatment with the gels and storage in saliva was conducted seven times. Finally, the samples were assessed for KOH-soluble and structurally bound fluoride. RESULTS: Only the enamel samples treated with the fluoridated bleaching gel (group B) and with the amine fluoride gel (group C) exhibited significant fluoride acquisition. Thereby, both gels showed significantly lower uptake in series 1 as compared to series 2. Both KOH-soluble and structurally bound fluoride acquisition was significantly higher in group C than in group B. CONCLUSION: Treatment with a carbamide peroxide gel supplemented with amine fluoride causes less fluoride acquisition in enamel than a pure amine fluoride gel. Under the conditions of the study, it is assumed that carbamide peroxide seems to influence enamel fluoride uptake. PMID- 16472905 TI - Association between the scores on the general health questionnaire-28 and the saliva levels of 3-methoxy-4-hydroxyphenylglycol in normal volunteers. AB - To access the saliva level of 3-methoxy-4-hydroxy-phenylglycol (sMHPG) as an index of mental health in normal volunteers, we investigated the relationship between the sMHPG and the scores on the general health questionnaire-28 (GHQ-28). A total of 270 normal volunteers answered the GHQ-28 and the sMHPG levels were determined. The sMHPG levels in women and men were comparable. There was a significant negative correlation between the social dysfunction score on the GHQ 28 and sMHPG levels in women (P=0.0035), but not in men. Moreover, the sMHPG levels also correlated with the total GHQ-28 score (P=0.0205), the anxiety and the insomnia score (P=0.0306) in women. These data indicate a high social dysfunction score on the GHQ-28 to be associated with a reduced noradrenergic neuronal tone thus possibly reflecting psychomotor retardation in women. PMID- 16472906 TI - Extending animal models of fear conditioning to humans. AB - A goal of fear and anxiety research is to understand how to treat the potentially devastating effects of anxiety disorders in humans. Much of this research utilizes classical fear conditioning, a simple paradigm that has been extensively investigated in animals, helping outline a brain circuitry thought to be responsible for the acquisition, expression and extinction of fear. The findings from non-human animal research have more recently been substantiated and extended in humans, using neuropsychological and neuroimaging methodologies. Research across species concur that the neural correlates of fear conditioning include involvement of the amygdala during all stages of fear learning, and prefrontal areas during the extinction phase. This manuscript reviews how animal models of fear are translated to human behavior, and how some fears are more easily acquired in humans (i.e., social-cultural). Finally, using the knowledge provided by a rich animal literature, we attempt to extend these findings to human models targeted to helping facilitate extinction or abolishment of fears, a trademark of anxiety disorders, by discussing efficacy in modulating the brain circuitry involved in fear conditioning via pharmacological treatments or emotion regulation cognitive strategies. PMID- 16472907 TI - Thermal behavior of novel non-sonicated arsonolipid-containing liposomes. AB - The thermal properties of novel arsonolipid-containing liposomes in PBS pH 7.4 and in water in absence and presence of Ca(2+) ions are reported. Liposomes composed of arsonolipids with different acyl chains (C(12), C(16) and C(18)) were prepared by the one step method. Microcalorimetry results showed that (i) the thermotropic transitions of arsonoliposomes (in PBS, pH 7.4, and in water) increase as a function of arsonolipid fatty acyl chain length, (ii) arsonoliposomes of long fatty acyl chain arsonolipids (C(16) and C(18)) showed higher enthalpy and transition temperature in the buffer compared to those observed in water (for arsonoliposomes of C(12)-fatty acyl chain arsonolipid, the order was reversed which might be attributed to their different structure), and (iii) the presence of 2 mM CaCl(2) has more pronounced effects on the thermal properties of arsonoliposomes in distilled water than in buffer, which suggests that the ionic strength of the dispersion medium plays an important role in determining the thermal properties of arsonoliposomes. PMID- 16472908 TI - Transgenic RNAi: a tool to study testis-specific genes. PMID- 16472909 TI - No association of the Arg51Gln and Leu72Met polymorphisms of the ghrelin gene with anorexia nervosa or bulimia nervosa. AB - Genetic factors likely contribute to the biological vulnerability to anorexia nervosa (AN) and bulimia nervosa (BN). We investigated whether the Arg51Gln and/or the Leu72Met gene polymorphisms of the human ghrelin, a peptide involved in the regulation of eating behavior, were associated to AN and/or BN. Two hundred-ninety-two Caucasian women (114 with BN, 59 with AN and 119 healthy controls) participated into the study. No significant differences were found in the frequencies of the Arg51Gln and the Leu72Met ghrelin gene variants among patients with AN or BN and healthy controls. Moreover, no significant differences emerged in eating-related phenotypic variables between patients carrying the Leu72Met genotype as compared to those with the Leu72Leu genotype. These results suggest that the Arg51Gln and the Leu72Met polymorphisms of the human ghrelin gene do not contribute to the genetic susceptibility to AN and BN. PMID- 16472910 TI - Association between dopamine D4 receptor (DRD4) exon III polymorphism and Neuroticism in the Japanese population. AB - The association between the dopamine D4 receptor (DRD4) exon III polymorphism and personality trait of novelty seeking (NS) has been studied intensively. In the Japanese population, the results of the previous studies did not always coincide. In the present study, we investigated the association between the polymorphism and personality traits evaluated by using the Revised NEO Personality Inventory (NEO PI-R) and State-Trait Anxiety Inventory (STAI) in 196 Japanese subjects. A meta-analysis of the present and previous Japanese studies was also conducted regarding NS. As a result, significant association was observed between the polymorphism and personality traits evaluated by using NEO PI-R as a whole (p=0.022, MANCOVA). Subsequent analyses showed a significant association between short alleles (2-4 repeats) and higher scores for Neuroticism or its subscales, Anxiety, Depression, and Vulnerability (p=0.015, 0.039, 0.021, and 0.008, respectively, uncorrected). No other significant difference in the scores for NEO PI-R was observed in the subsequent analyses. Significant association was also observed between the polymorphism and scores for STAI as a whole (p=0.004, MANCOVA). Subsequent analyses did not show significant association, although a weak trend for the relation between the genotype consisting of short alleles and Trait Anxiety was observed (p=0.10, uncorrected). The meta-analysis showed no significant association between the polymorphism and NS. Thus, the present study suggested the association between the short allele of the DRD4 exon III polymorphism and personality trait of Neuroticism in Japanese subjects. PMID- 16472911 TI - Early reduction of NeuN antigenicity induced by soman poisoning in mice can be used to predict delayed neuronal degeneration in the hippocampus. AB - The neuronal nuclei (NeuN) antigen is increasingly being used as a specific marker to identify neuronal cell loss under various pathological conditions. However, recent studies pointed out that a decrease in NeuN labeling could also be due to the reduction of protein expression level or loss of antigenicity and this was not necessarily related to neuronal cell disappearance. We also investigated the presence of damaged neurons, the loss of NeuN immunoreactivity and the level of NeuN protein in the brain hippocampus of mice subjected to soman poisoning (1.2 LD50 of soman). Damaged neurons were detected using hemalun phloxin (H&P) and Fluoro-Jade B (FJB) staining on brain sections. NeuN immunohistochemistry was also performed on adjacent brain sections and NeuN protein level quantified by Western blot analysis. One and eight days after soman exposure, about 49% of hippocampal neurons were damaged, as assessed by H&P or FJB staining. NeuN immunohistochemistry indicated that all these damaged neurons were deprived of NeuN immunoreactivity. Using Western blot analysis, we proved that loss of NeuN immunoreactivity in degenerating neurons was due to reduced NeuN antigenicity rather than a fall in protein expression level. In this study, we discuss the potential use of NeuN immunohistochemistry as a good biomarker to predict delayed neuronal degeneration in the rodent hippocampus after various brain injuries. PMID- 16472912 TI - Bilateral phase entrainment by movement-elicited afference contributes equally to the stability of in-phase and antiphase coordination. AB - This study examined if and how phase entrainment by movement-related afference-- induced by passive movements of the contralateral hand-- contributes to the coordinative stability of rhythmic bimanual movements. The results revealed that phase-shifted passive movements of the dominant or the nondominant hand induced similar entrainment effects on the active movements of the contralateral hand. In terms of the phase relations between the hands only two attractors of equal strength were present, corresponding to relative phase shifts of 0 degrees and 180 degrees , respectively, i.e., to in-phase and antiphase coordination. The attractors were separated by two repellors located at relative phase shifts of +90 degrees and -90 degrees . EMG analysis indicated that the entrainment effects indeed resulted from contralateral afference, because no related changes in muscle activation were observed in the passively moved hand. It was concluded that phase entrainment by contralateral afference contributed equally to the stable performance of the bimanual in-phase and antiphase patterns, thereby enhancing the stability of these patterns relative to other phase relations between the limbs. PMID- 16472914 TI - People in pain. PMID- 16472913 TI - Mitochondrial electron transport in models of neuropathic and inflammatory pain. AB - Although peripheral nerve function is strongly dependent on energy stores, the role of the mitochondrial electron transport chain, which drives ATP synthesis, in peripheral pain mechanisms, has not been examined. In models of HIV/AIDS therapy (dideoxycytidine), cancer chemotherapy (vincristine), and diabetes (streptozotocin)-induced neuropathy, inhibitors of mitochondrial electron transport chain complexes I, II, III, IV, and V significantly attenuated neuropathic pain-related behavior in rats. While inhibitors of all five complexes also attenuated tumor necrosis factor alpha-induced hyperalgesia, they had no effect on hyperalgesia induced by prostaglandin E2 and epinephrine. Two competitive inhibitors of ATP-dependent mechanisms, adenosine 5'-(beta,gamma imido) triphosphate and P1,P4-di(adenosine-5') tetraphosphate, attenuated dideoxycytidine, vincristine, and streptozotocin-induced hyperalgesia. Neither of these inhibitors, however, affected tumor necrosis factor alpha, prostaglandin E2 or epinephrine hyperalgesia. These experiments demonstrate a role of the mitochondrial electron transport chain in neuropathic and some forms of inflammatory pain. The contribution of the mitochondrial electron transport chain in neuropathic pain is ATP dependent. PMID- 16472915 TI - Patient-perceived satisfactory improvement (PPSI): interpreting meaningful change in pain from the patient's perspective. AB - The assessment of clinically meaningful changes in patient-reported pain has become increasingly important when interpreting results of clinical studies. However, proposed response criteria, such as the minimal clinically important difference, do not correspond with the growing need for information on truly meaningful, individual improvements. The aim of the present study was to investigate satisfactory improvements in pain from the patient's perspective. Data were collected in a 2-week prospective study of 181 arthritis patients treated with a local corticosteroid injection. Baseline and follow-up pain were assessed on 100mm visual analogue scales for pain intensity (VAS-PI). At baseline, patients also marked a hypothetical level on a VAS-PI representing a satisfactory improvement in pain. Patient-perceived satisfactory improvement (PPSI) was constructed using a 5-point categorical rating of change scale at follow-up as the anchor. PPSI was associated with a minimal reduction of 30mm or 55% on the VAS-PI. Since absolute change in pain associated with satisfactory improvement proved highly dependent on baseline pain, percent change scores performed better in classifying improved patients. The 55% threshold for satisfactory improvement was consistent over the course of treatment and reasonably consistent across groups of patients. Our data suggest that PPSI is a clinically relevant and stable concept for interpreting truly meaningful improvements in pain from the individual perspective. PMID- 16472916 TI - The importance of symptom duration in determining prognosis. AB - Symptom duration is integral to clinical and epidemiological research on pain. It is widely used for sample selection and commonly assessed in clinical practice. However, there has been little specific investigation of the link between duration and outcome. This work aimed to examine the association of episode duration with symptoms and clinical course in primary care consulters with low back pain (LBP). In a cohort of general practice LBP consulters, 619 patients returned two or more questionnaires during a 12-month period. LBP episode duration at baseline consultation was defined by time since their last pain-free month. Associations of duration with pain, disability and psychological status at baseline were examined. Survival analysis was used to assess the association between episode duration and time taken for disability to improve by 30%. At baseline, increasing episode duration was associated with worse pain, disability and psychological status (p < 0.001), but there were no differences between people with more or less than 3 months of pain. People with 3 years' or more duration at baseline took significantly longer to improve than those with shorter duration (adjusted hazard ratio 1.57, 95% confidence interval 1.27-1.95). In conclusion, memory of LBP episode duration is associated with pain, disability and psychological status, and is an independent predictor of time to improvement. There are important differences between people who recall more or less than 3 years' duration. Mechanisms for these associations are poorly understood, but this research suggests that duration itself is an important focus for research. PMID- 16472917 TI - Glucocorticoid inhibition of vascular abnormalities in a tibia fracture rat model of complex regional pain syndrome type I. AB - Tibia fracture in rats evokes chronic hindpaw warmth, spontaneous extravasation, edema, allodynia, and periarticular bone loss, a syndrome resembling complex regional pain syndrome type I (CRPS I). Glucocorticoids such as methylprednisolone (MP) are probably effective analgesic and anti-edematous agents in patients suffering from CRPS and this study examined the effects of chronic MP treatment in the rat CRPS I model. Bilateral hindpaw thickness, temperature, and nociceptive thresholds were determined, and the hindlimb bone density was measured using dual-energy X-ray absorptiometry (DXA). Spontaneous cutaneous extravasation and substance P infusion evoked extravasation were determined using an Evans blue vascular permeability assay. After baseline testing, the distal tibia was fractured and the hindlimb casted for 4 weeks. At 2 weeks post-fracture MP infusion was started (1 mg/kg/day for 28 days). The rats were retested at 4, 6, and 8 weeks post-fracture. Hindpaw edema and warmth after fracture were reversed by MP infusion and these effects persisted after discontinuing treatment. Furthermore, there was an increase in spontaneous protein extravasation and an enhanced substance P evoked extravasation and edema response in the hindpaw at 4 weeks that was inhibited by MP infusion. Glucocorticoid treatment had no effect on the allodynia, hindpaw unweighting, or the periarticular bone loss observed after tibia fracture. We postulate that post junctional facilitation of substance P signaling contributes to the hindpaw warmth, edema, and the enhanced spontaneous protein extravasation observed in this CRPS I model, and that the anti-edematous effects of glucocorticoid treatment are due to inhibition of post-junctional neuropeptide signaling. PMID- 16472918 TI - Pharmacological assessment of the freezing, antinociception, and exploratory behavior organized in the ventrolateral periaqueductal gray. AB - Opioid and serotonergic mechanisms of the ventrolateral periaqueductal gray (vlPAG) are recruited by conditioned freezing and antinociception. However, it is unclear whether freezing and antinociception induced by stimulation of the vlPAG are interrelated. To address this issue we looked at the effects of the opioid antagonist naltrexone, the 5-HT2 antagonist ketanserin, and the benzodiazepine agonist midazolam injected into the vlPAG on the freezing and antinociception induced by electrical stimulation of this region. This antinociception was evaluated by the tail-flick and formalin tests. To further characterize the involvement of the vlPAG in unconditioned fear, the effects of intra-vlPAG injections of midazolam on the exploratory behavior were also assessed in independent groups of rats submitted to the elevated plus-maze test (EPM). The data obtained showed that: (i) electrical stimulation of the vlPAG causes freezing blocked by midazolam but not by naltrexone and ketanserin; (ii) antinociception generated at the level of the vlPAG is inhibited by naltrexone, ketanserin, and midazolam; (iii) activation of benzodiazepine-mediated mechanisms in the vlPAG increased the exploratory behavior of rats in the closed arms but not the avoidance behavior of open arms of the EPM. Thus, freezing and antinociception generated in the vlPAG are dissociated pharmacologically. Whereas antinociception is a multimediated process sensitive to naltrexone, ketanserin, and midazolam, the freezing induced by vlPAG stimulation was reversed only by the benzodiazepine compound. As injections of midazolam into the vlPAG do not cause anxiolytic effects in the EPM, the aversive stimuli inherent of this test seem to bypass the vlPAG. PMID- 16472919 TI - Current evidence for a genetic modulation of the response to analgesics. PMID- 16472920 TI - The productivity effects of cattle tick (Boophilus microplus) infestation on cattle, with particular reference to Bos indicus cattle and their crosses. AB - In response to uncertainty among cattle producers in Australia regarding the need to treat Bos indicus and B. indicus crossbreeds, the scientific literature relating to the productivity effects of Boophilus microplus on cattle of all breeds was reviewed. Estimates of the mean effect of each engorging tick (damage coefficient, d) were made from a simple analysis of the reported data. On average, each engorging female tick is responsible for the loss of 1.37 +/- 0.25 g bodyweight in B. taurus cattle. The comparable value for B. taurusxB. indicus cattle is 1.18 +/- 0.21 g/engorging tick. These values were not statistically significantly different, indicating that if a threshold approach to tick control were taken, then the threshold number of standard ticks would be the same regardless of cattle genotype. No studies provided useable estimates of the effect of tick infestation on pure B. indicus cattle. An economic threshold for treatment, below which acaricide application is not beneficial, can be predicted, using known values for the cost of acaricide application and the price of beef. However, the application of a threshold approach to control has not been embraced by government advisers and runs contrary to the accepted principals of strategic control programs. PMID- 16472921 TI - An examination of the prevalence of and risk factors for shedding of Cryptosporidium spp. and Giardia spp. in cows and calves from western Canadian cow-calf herds. AB - The primary objectives of this study were to describe the prevalence and risk factors for Cryptosporidium spp. and Giardia spp. infection in cows and calves during the calving season in western Canadian cow-calf herds. Through the calving season of 2002, fresh fecal samples were collected from 560 beef cows and 605 calves in western Canada. Feces were examined for the presence of Cryptosporidium spp. and Giardia spp. using a quantitative sucrose gradient immunoflourescent antibody test. Samples were collected from mature cows on 59 farms and from calves on 100 farms. Only 1.1% (5/560) of the cows were positive for Cryptosporidium spp. whereas 3.1% (19/605) of the calves were positive. Prevalence for Giardia spp. was much higher; Giardia spp. was detected in 17.0% (95/560) of the cow and 22.6% (137/605) of calf fecal samples. Data describing herd management practices, treatment and disease history, age, gender, breed and fecal consistency were gathered to assess potential risk factors associated with shedding. The association between the risk of shedding and average precipitation from December to June and ecological region were also evaluated. Risk factors for infection with Cryptosporidium spp. in either cows or calves could not be evaluated because the multilevel model would not converge due to the relatively low prevalence of the organism in this sample. The prevalence for Giardia spp. was sufficient to explore potential risk factors in both cows and calves. No risk factors were identified for Giardia spp. in beef cows following calving. After the construction of a multivariable model, the only significant predictors for Giardia spp. presence in beef calves was dam age and calf age. Calves born to 2 year-old heifers were 2.3 (95% CI, 1.09-5.06; P = 0.031) times more likely to be shedding Giardia spp. then calves born to cows that were 4-10 years of age. Calves that were 9-18 days of age and calves that were > 18 days of age were 22.4 (95% CI, 5.88-88.18; P < 0.001) and 150 (95% CI, 39.72-603.19; P < 0.001) times more likely, respectively, to be shedding Giardia spp. than calves < or = 4 days of age. PMID- 16472922 TI - A modified larval migration assay for detection of resistance to macrocyclic lactones in Haemonchus contortus, and drug screening with Trichostrongylidae parasites. AB - We have developed a modified migration assay system in 96-well plate format which is able to detect resistance to the macrocyclic lactone group of drugs in Haemonchus contortus. The assay involves exposure of infective stage larvae to drug for a 24 h period, then counting the numbers of larvae that are able to migrate through an agar and filter mesh system over a further 48 h. The agar barrier greatly increased the sensitivity of the assay for resistance detection compared to use of filter mesh alone. The assay was able to detect the presence of 10% resistant worms in an otherwise susceptible background. However, the assay was ineffective with Trichostrongylus colubriformis and Ostertagia circumcincta indicating that its usefulness for field monitoring will be restricted to situations where H. contortus is of most significance. A small-scale drug screening exercise showed that the assay identifies some anthelmintic activities distinct from those identified by larval development assays. The assay therefore also has a potential role in drug discovery programmes in screening for new anthelmintics. PMID- 16472923 TI - PCR detection of Neospora caninum, Toxoplasma gondii and Encephalitozoon cuniculi in brains of wild carnivores. AB - Neospora caninum, Toxoplasma gondii and Encephalitozoon cuniculi are important pathogens with affinity to the central nervous system of many animals. 240 brains of wild carnivores were examined by PCR-based diagnosis. The presence of N. caninum DNA was confirmed in 4.61% (7/152) red foxes (Vulpes vulpes). DNA of T. gondii was found in 4.92% (3/61) martens (Martes sp.) and in 1.32% (2/152) red foxes. DNA of E. cuniculi was determined in 3.28% (2/61) martens and in one examined European otter (Lutra lutra). There were no co-infections found. These results provide the first evidence of E. cuniculi in the European otter, the first report of N. caninum in foxes in the Czech Republic and confirm the presence of T. gondii in wild carnivores in the Czech Republic. PMID- 16472924 TI - Effectiveness of stereotactic radiosurgery alone or in combination with whole brain radiotherapy compared to conventional surgery and/or whole brain radiotherapy for the treatment of one or more brain metastases: a systematic review and meta-analysis. AB - PURPOSE: To assess the effectiveness of SRS alone or in combination with WBRT compared to surgery and/or WBRT in prolonging survival and improving the quality of-life and functional status of patients with brain metastases. METHODS AND MATERIALS: A meta-analysis of randomized controlled trials and concurrent cohort studies examining SRS versus SRS + WBRT, SRS versus WBRT +/- surgical resection, SRS versus surgical resection only, or SRS + WBRT versus WBRT was conducted. Trial registers, bibliographic databases, and reference lists from selected studies and recent issues of relevant journals were searched. Neuro-oncology specialists were also contacted. All studies were analyzed independently by two reviewers, applying validated critical appraisal techniques. RESULTS: The review identified three RCTs and one cohort study. Among patients with multiple metastases, no difference in survival between those treated with WBRT + SRS and those treated with WBRT was found. However, in patients with one metastasis, a statistically significant difference, favoring those treated with WBRT + SRS, was observed. Regarding local tumor control at 24 months, rates were significantly higher in the WBRT + SRS treatment arm, regardless of the number of metastases. CONCLUSIONS: Adding SRS to WBRT improves survival in patients with one brain metastasis. Combining SRS and WBRT improves local tumour control and functional independence in all patients. PMID- 16472925 TI - Hormonal therapy for male breast cancer: A different approach for a different disease. AB - Male breast cancer (MBC) is on the rise in the United States [Surveillance, Epidemiology, and End Results (SEER) Program () SEER Stat Database: Incidence SEER 9 Regs Public-Use; November 2004 submission (1973-2002), National Cancer Institute, DCPPS, Surveillance Research Program, Cancer Statistics Branch, released April 2005, based on the November 2004 submission]; however mortality due to MBC has not changed unlike in its female counterpart [American Cancer Society: Cancer facts and figures 2005. Atlanta (GA): American Cancer Society; 2005]. The rarity of MBC has precluded major progress in the understanding and treatment of this disease. Treatment has often been extrapolated from female breast cancer (FBC) despite distinct clinicopathologic features between the two entities, especially with regards to the role of male hormones and estrogens in this disease. Also, it is uncertain if hormone receptor positive tumors carry the same prognostic implication in MBC as in the female disease. Hormonal therapy has been the mainstay of treatment in MBC with tamoxifen the front-line drug. The role of the newer generation aromatase inhibitors has not been well defined but they are being used in clinical practice for the treatment of MBC, based on accepted data for women with the disease. This commentary focuses on the major hormonal differences between male and female breast cancer that would suggest the need to explore different treatment strategies if significant advances are to be made in the understanding and treatment of this distinct disease. PMID- 16472926 TI - Cigarette smoking induces heat shock protein 70 kDa expression and apoptosis in rat brain: Modulation by bacoside A. AB - Cigarette smoking is associated with the development of several diseases and antioxidants play a major role in the prevention of smoking-related diseases. Apoptosis is suggested as a possible contributing factor in the pathogenesis of smoking-induced toxicity. Therefore the present study was designed to investigate the influence of chronic cigarette smoke exposure on apoptosis and the modulatory effect of bacoside A (triterpenoid saponin isolated from the plant Bacopa monniera) on smoking-induced apoptosis in rat brain. Adult male albino rats of Wistar strain were exposed to cigarette smoke and simultaneously administered with bacoside A (10 mg/kg b.w./day, orally) for a period of 12 weeks. Expression of brain hsp70 was analyzed by Western blotting. Apoptosis was identified by DNA fragmentation, terminal deoxynucleotidyl transferase-mediated deoxy uridine triphosphate nick end labeling (TUNEL) staining and transmission electron microscopy. The results showed that exposure to cigarette smoke induced hsp70 expression and apoptosis as characterized by DNA laddering, increased TUNEL positive cells and ultrastructural apoptotic features in the brain. Administration of bacoside A prevented expression of hsp70 and neuronal apoptosis during cigarette smoking. We speculate that apoptosis may be responsible for the smoking-induced brain damage and bacoside A can protect the brain from the toxic effects of cigarette smoking. PMID- 16472928 TI - Modifications of local cerebral glucose utilization during circadian food anticipatory activity. AB - Food-anticipatory activity that animals express before a daily timed meal is considered as the behavioral output of a feeding-entrainable oscillator whose functional neuroanatomy is still unknown. In order to identify the possible brain areas involved in that timing mechanism, we investigated local cerebral metabolic rate for glucose during food-anticipatory activity produced either by a 4-h daily access to food starting 4 h after light onset or by a hypocaloric feeding provided at the same time. Local cerebral metabolic rate for glucose measured by the labeled 2-[(14)C]-deoxyglucose technique was quantified in 40 structures. In both groups of food-restricted rats, three brain regions (the nucleus of the solitary tract, the cerebellar cortex and the medial preoptic area) showed a decrease in local cerebral metabolic rate for glucose, compared with control ad libitum animals. In addition, only one structure, the paraventricular thalamic nucleus, was affected by temporal restricted feeding, and not by hypocaloric feeding, compared with ad libitum rats. By contrast, three brain regions, i.e. the intergeniculate leaflets, the paraventricular hypothalamic and the arcuate nuclei, showed specifically metabolic decreases during anticipation of hypocaloric feeding, and not during anticipation of temporal restricted feeding, compared with the ad libitum group. Expression of food-anticipatory activity appears to be regulated by an integrated neural circuit of brainstem and hypothalamic pathways, with hypocaloric feeding involving more extensive forebrain areas than temporal restricted feeding. PMID- 16472927 TI - Presynaptic alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors modulate release of inhibitory amino acids in rat spinal cord dorsal horn. AB - Local inhibition within the spinal cord dorsal horn is mediated by the neurotransmitters GABA and glycine and strongly influences nociceptive and temperature signaling. Alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors are expressed by inhibitory interneurons and have been shown to modulate GABA release in other regions of the CNS. In the spinal cord, there is morphological evidence for presynaptic AMPA receptor subunits in GABAergic dorsal horn neurons, but functional data are lacking. To determine if AMPA receptors are indeed functional at presynaptic terminals of inhibitory neurons, we recorded evoked and miniature inhibitory postsynaptic currents (mIPSPs) in the superficial dorsal horn of the rat spinal cord. We show that AMPA receptor activation enhances spontaneous release of inhibitory amino acids in the presence of tetrodotoxin onto both lamina II neurons and NK1 receptor-expressing (NK1R+) lamina I neurons. This effect is sensitive to the concentration of extracellular Ca2+, yet is not fully blocked in most neurons in the presence of Cd2+, suggesting possible Ca2+ entry through AMPA receptors. Postsynaptic Ca2+ elevation is not required for these changes. AMPA-induced increases in mIPSP frequency are also seen in more mature dorsal horn neurons, indicating that these receptors may play a role in nociceptive processing in the adult. In addition, we have observed AMPA-induced depression of evoked release of GABA and glycine onto lamina I NK1R+ neurons. Taken together these data support a role for presynaptic AMPA receptors in modulating release of GABA and glycine in the superficial dorsal horn. Because inhibition in the dorsal horn is important for controlling pain signaling, presynaptic AMPA receptors acting to modulate the inhibitory inputs onto dorsal horn neurons would be expected to impact upon pain signaling in the spinal cord dorsal horn. PMID- 16472930 TI - Blockade of arachidonic acid pathway induces sprouting in the adult but not in the neonatal uncrossed retinotectal projection. AB - The uncrossed retinotectal projection of rats undergoes extensive axonal elimination and subsequent growth of axonal arbors in topographically appropriate territories within the first two/three postnatal weeks. Nitric oxide has been implicated in development and stabilization of synapses in the retinotectal pathway since blockade of nitric oxide synthesis disrupts the normal pattern of retinal innervation in subcortical nuclei. The present work investigated the role of arachidonic acid pathway in the development and maintenance of ipsilateral retinotectal axons. We also investigated the role of this retrograde messenger in the modulation of plasticity that follows retinal lesions in the opposite eye. Pigmented rats received systemic treatment with quinacrine, a phospholipase A2 inhibitor, indomethacin, a cyclooxygenase inhibitor, nordihydroguaiaretic acid, a 5-lipoxygenase inhibitor or vehicle during 4-8 days at various postnatal ages. Rats given a unilateral temporal retinal lesion were treated with either quinacrine or vehicle during the same period. For anterograde tracing of ipsilateral retinal projections, animals received intraocular injections of horseradish peroxidase. Before the third postnatal week no difference was observed in the laminar or topographic organization of the ipsilateral retinotectal projection between vehicle and treated rats in either normal or lesion conditions. After the third postnatal week, however, systemic blockade of phospholipase A2 or 5-lipoxygenase, but not cyclooxygenase induced sprouting of uncrossed axons throughout the collicular visual layers in unoperated rats. In retinal lesion groups, phospholipase A2 blockade increased the sprouting of uncrossed intact axons to the collicular surface in the same period. The results suggest that arachidonic acid or lipoxygenase metabolites play a role in the maintenance of the retinotectal synapses after the critical period and that the blockade of the arachidonic acid pathway induces reactive sprouting of retinal axons late in development. PMID- 16472929 TI - The roles of different subtypes of opioid receptors in mediating the nucleus submedius opioid-evoked antiallodynia in a neuropathic pain model of rats. AB - Previous studies have indicated that thalamic nucleus submedius is involved in opioid-mediated antinociception in tail flick test and formalin test. The current study examined the effects of opioids microinjected into the thalamic nucleus submedius on the allodynia developed in neuropathic pain model rats, and determined the roles of different subtypes of opioid receptors in the thalamic nucleus submedius opioid-evoked antiallodynia. The allodynic behaviors induced by L5/L6 spinal nerve ligation were assessed by mechanical (von Frey filaments) and cold (4 degrees C plate) stimuli. Morphine (1.0, 2.5, and 5.0 microg) microinjected into the thalamic nucleus submedius contralateral to the nerve injury paw produced a dose-dependent inhibition of the mechanical and cold allodynia, and these effects were reversed by microinjection of the non-selective opioid receptor antagonist naloxone (1.0 microg) into the same site. Microinjection of endomorphin-1 (5.0 microg), a highly selective mu-opioid receptor agonist, and [D-Ala2, D-Leu5]-enkephalin (10 microg), a delta-/mu-opioid receptor agonist, also inhibited the allodynic behaviors, and these effects were blocked by selective mu-opioid receptor antagonist beta-funaltrexamine hydrochloride (3.75 microg). However, the [D-Ala2, D-Leu5]-enkephalin-evoked antiallodynic effects were not influenced by the selective delta-opioid receptor antagonist naltrindole (5.0 microg). Microinjection of the selective kappa receptor agonist spiradoline mesylate salt (100 microg) into the thalamic nucleus submedius failed to alter the allodynia induced by spinal nerve ligation. These results suggest that the thalamic nucleus submedius is involved in opioid-evoked antiallodynia which is mediated by mu- but not delta- and kappa-opioid receptor in the neuropathic pain model rats. PMID- 16472932 TI - GHB use among gay and bisexual men. AB - The recreational use of gamma-hydroxybutyrate (GHB) has been relatively understudied, despite its popularity in gay communities. We examined the use of GHB in a sample of 450 club drug using gay and bisexual men. Of these, 29% indicated use of the substance in the recent past. GHB users were similar to those in the sample who reported no use along key demographic factors, although GHB users were more likely to identify as gay than bisexual and were slightly older. Poly-drug use was common, with close to half of GHB users combining with methamphetamine, MDMA, or ketamine; approximately one quarter also used GHB with alcohol. Participants reported that GHB was often used at nightclubs, circuit parties, sex parties, and sex clubs, with HIV-positive men more likely to use the substance in sexual contexts. Use of GHB is common among a certain subset of gay men despite warnings within the community about the potentially fatal effects of the substance, suggesting that more effort be given to educate drug using gay men about GHB. PMID- 16472931 TI - An overview of medications for the treatment of alcohol withdrawal and alcohol dependence with an emphasis on the use of older and newer anticonvulsants. AB - There is a growing interest in the development of new pharmacological tools for treating alcohol withdrawal and dependence. A number of anticonvulsants including valproate and carbamazepine have been shown to be safe and effective alternatives to benzodiazepines for treating alcohol withdrawal. These agents are relatively safe, are free from demonstrated abuse liability, and do not usually potentiate the psychomotor and cognitive effects of alcohol. For the treatment of alcohol dependence, there is a growing literature on the utility of medications that have neurochemical effects at opioid, serotonergic, GABAergic, and glutamate receptors. Furthermore, as a class of medication, there appears to be a growing interest in investigating the utility of novel anticonvulsants such as topiramate for the treatment of alcohol dependence. PMID- 16472934 TI - Preliminary results of a phase I/II study of simultaneous modulated accelerated radiotherapy for nondisseminated nasopharyngeal carcinoma. AB - PURPOSE: To present preliminary results of intensity-modulated radiotherapy (IMRT) with the simultaneous modulated accelerated radiotherapy (SMART) boost technique in patients with nasopharyngeal carcinoma (NPC). METHODS AND MATERIALS: Twenty patients who underwent IMRT for nondisseminated NPC at the Asan Medical Center between September 2001 and December 2003 were prospectively evaluated. Intensity-modulated radiotherapy was delivered with the "step and shoot" SMART technique at prescribed doses of 72 Gy (2.4 Gy/day) to the gross tumor volume, 60 Gy (2 Gy/day) to the clinical target volume and metastatic nodal station, and 46 Gy (2 Gy/day) to the clinically negative neck region. Eighteen patients also received cisplatin once per week. RESULTS: The median follow-up period was 27 months. Nineteen patients completed the treatment without interruption; the remaining patient interrupted treatment for 2 weeks owing to severe pharyngitis and malnutrition. Five patients (25%) had Radiation Therapy Oncology Group Grade 3 mucositis, whereas 9 (45%) had Grade 3 pharyngitis. Seven patients (35%) lost more than 10% of their pretreatment weight, whereas 11 (55%) required intravenous fluids and/or tube feeding. There was no Grade 3 or 4 xerostomia. All patients showed complete response. Two patients had distant metastases and locoregional recurrence, respectively. CONCLUSION: Intensity-modulated radiotherapy with the SMART boost technique allows parotid sparing, as shown clinically and by dosimetry, and might also be more effective biologically. A larger population of patients and a longer follow-up period are needed to evaluate ultimate tumor control and late toxicity. PMID- 16472933 TI - Practice guideline for the performance of therapy with unsealed radiopharmaceutical sources. PMID- 16472935 TI - Prognostic index score and clinical prediction model of local regional recurrence after mastectomy in breast cancer patients. AB - PURPOSE: To develop clinical prediction models for local regional recurrence (LRR) of breast carcinoma after mastectomy that will be superior to the conventional measures of tumor size and nodal status. METHODS AND MATERIALS: Clinical information from 1,010 invasive breast cancer patients who had primary modified radical mastectomy formed the database of the training and testing of clinical prognostic and prediction models of LRR. Cox proportional hazards analysis and Bayesian tree analysis were the core methodologies from which these models were built. To generate a prognostic index model, 15 clinical variables were examined for their impact on LRR. Patients were stratified by lymph node involvement (<4 vs. >or =4) and local regional status (recurrent vs. control) and then, within strata, randomly split into training and test data sets of equal size. To establish prediction tree models, 255 patients were selected by the criteria of having had LRR (53 patients) or no evidence of LRR without postmastectomy radiotherapy (PMRT) (202 patients). RESULTS: With these models, patients can be divided into low-, intermediate-, and high-risk groups on the basis of axillary nodal status, estrogen receptor status, lymphovascular invasion, and age at diagnosis. In the low-risk group, there is no influence of PMRT on either LRR or survival. For intermediate-risk patients, PMRT improves LR control but not metastases-free or overall survival. For the high-risk patients, however, PMRT improves both LR control and metastasis-free and overall survival. CONCLUSION: The prognostic score and predictive index are useful methods to estimate the risk of LRR in breast cancer patients after mastectomy and for estimating the potential benefits of PMRT. These models provide additional information criteria for selection of patients for PMRT, compared with the traditional selection criteria of nodal status and tumor size. PMID- 16472936 TI - Protection against radiation-induced oxidative stress in cultured human epithelial cells by treatment with antioxidant agents. AB - PURPOSE: To evaluate the protective effects of antioxidant agents against space radiation-induced oxidative stress in cultured human epithelial cells. METHODS AND MATERIALS: The effects of selected concentrations of N-acetylcysteine, ascorbic acid, sodium ascorbate, co-enzyme Q10, alpha-lipoic acid, l selenomethionine, and vitamin E succinate on radiation-induced oxidative stress were evaluated in MCF10 human breast epithelial cells exposed to radiation with X rays, gamma-rays, protons, or high mass, high atomic number, and high energy particles using a dichlorofluorescein assay. RESULTS: The results demonstrated that these antioxidants are effective in protecting against radiation-induced oxidative stress and complete or nearly complete protection was achieved by treating the cells with a combination of these agents before and during the radiation exposure. CONCLUSION: The combination of antioxidants evaluated in this study is likely be a promising countermeasure for protection against space radiation-induced adverse biologic effects. PMID- 16472938 TI - Modeling radiation dosimetry to predict cognitive outcomes in pediatric patients with CNS embryonal tumors including medulloblastoma. AB - PURPOSE: Model the effects of radiation dosimetry on IQ among pediatric patients with central nervous system (CNS) tumors. METHODS AND MATERIALS: Pediatric patients with CNS embryonal tumors (n = 39) were prospectively evaluated with serial cognitive testing, before and after treatment with postoperative, risk adapted craniospinal irradiation (CSI) and conformal primary-site irradiation, followed by chemotherapy. Differential dose-volume data for 5 brain volumes (total brain, supratentorial brain, infratentorial brain, and left and right temporal lobes) were correlated with IQ after surgery and at follow-up by use of linear regression. RESULTS: When the dose distribution was partitioned into 2 levels, both had a significantly negative effect on longitudinal IQ across all 5 brain volumes. When the dose distribution was partitioned into 3 levels (low, medium, and high), exposure to the supratentorial brain appeared to have the most significant impact. For most models, each Gy of exposure had a similar effect on IQ decline, regardless of dose level. CONCLUSIONS: Our results suggest that radiation dosimetry data from 5 brain volumes can be used to predict decline in longitudinal IQ. Despite measures to reduce radiation dose and treatment volume, the volume that receives the highest dose continues to have the greatest effect, which supports current volume-reduction efforts. PMID- 16472939 TI - Addressing glucose sensitivity measured by F-18 FDG PET in lung cancers for radiation treatment planning and monitoring. AB - PURPOSE: To address glucose sensitivity in lung cancers before and after radiation treatment (Tx). METHODS AND MATERIALS: Twelve patients were each studied with two pre-Tx positron emission tomography (PET) scans and 3 patients each with one post-Tx PET scan, with glucose concentration [Glc] and maximum standard uptake value (SUV) recorded. The pre-Tx glucose sensitivity, g from SUV1/SUV2= {[Glc]1/[Glc]2}g and Tx index, tau from SUVpost-Tx/SUVpre-Tx = {[Glc]post-Tx/[Glc]pre-Tx}tau was calculated by linear regression. Pre-Tx SUVs were corrected to post-Tx Glc with g (SUV'pre-Tx) for a pure Tx effect, R = ln(SUVpost-Tx/SUV'pre-Tx). RESULTS: There were no significant differences in SUV but [Glc] were different (96.4 +/- 10.9 vs. 88.3 +/- 10.5, p = 0.015) between two pre-Tx PET scans. Linear regression yielded g = -0.79 and tau = -1.78 to -2.41 (p < 0.0005 in all). The %DeltaSUV after Tx for 3 patients without vs. with g correction were different by -12%, 0%, and + 7%, suggesting varying effects from glucose. R values were also different and mean R (-0.81 +/- 0.38) was significantly different from zero (p = 0.03), consistent with successful Tx as confirmed by clinico-radiologic follow-up. CONCLUSIONS: The extra dimension of glucose sensitivity, g besides SUV incorporated in the combined Tx-derived tau may be a useful global Tx evaluation index even with differing [Glc]. PMID- 16472937 TI - Sustained long-term immune responses after in situ gene therapy combined with radiotherapy and hormonal therapy in prostate cancer patients. AB - PURPOSE: To explore long-term immune responses after combined radio-gene-hormonal therapy. METHODS AND MATERIALS: Thirty-three patients with prostate specific antigen 10 or higher or Gleason score of 7 or higher or clinical stage T2b to T3 were treated with gene therapy that consisted of 3 separate intraprostatic injections of AdHSV-tk on Days 0, 56, and 70. Each injection was followed by 2 weeks of valacyclovir. Intensity-modulated radiation therapy was delivered 2 days after the second AdHSV-tk injection for 7 weeks. Hormonal therapy was initiated on Day 0 and continued for 4 months or 2.3 years. Blood samples were taken before, during, and after treatment. Lymphocytes were analyzed by fluorescent antibody cell sorting (FACS). RESULTS: Median follow-up was 26 months (range, 4 48 months). The mean percentages of DR+CD8+ T cells were increased at all timepoints up to 8 months. The mean percentages of DR+CD4+ T cells were increased later and sustained longer until 12 months. Long-term (2.3 years) use of hormonal therapy did not affect the percentage of any lymphocyte population. CONCLUSIONS: Sustained long-term (up to 8 to 12 months) systemic T-cell responses were noted after combined radio-gene-hormonal therapy for prostate cancer. Prolonged use of hormonal therapy does not suppress this response. These results suggest the potential for sustained activation of cell-mediated immune responses against cancer. PMID- 16472940 TI - Formulation considerations for the development of medications with abuse potential. AB - The availability of increasingly sophisticated drug formulations and delivery devices has created new opportunities in drug development. These newer approaches can result in improved drug bioavailability, or they can alter key pharmacokinetic parameters in such a way as to decrease dosing interval, decrease variability, or blunt maximal concentrations that are associated with adverse events of particular concern. Special formulations or devices can also provide for easier or more convenient dosing in subpopulations of interest, such as children or the elderly. Although the key principles of abuse potential assessment and the underlying neurochemistry and pharmacology are relatively well understood, evaluation of the influences of drug formulation have received much less study. Because dose and formulation - and even the therapeutic indication - are refined over the course of development, it is usually difficult to conduct more than a cursory evaluation of the influence of formulation on the underlying abuse potential of the active pharmaceutical ingredient. Industrial sponsors would benefit from further research in areas of formulation science and pharmacokinetics that would improve the predictability of prescription drug abuse. In particular, validation of new pharmacokinetic parameters and standardization of methods to understand the consequences of product tampering could assist in gaining a better assessment of risk for controlled release formulations. Such methods could apply not only to innovator products, but also to those wishing to develop generic versions with similar pharmaceutical performance. This article proposes several factors that may be of use to industrial sponsors in making formulation choices for drugs with the potential for abuse. PMID- 16472941 TI - A novel discoidin domain receptor 1 (Ddr1) transcript is expressed in postmeiotic germ cells of the rat testis depending on the major histocompatibility complex haplotype. AB - The Ddr1 gene encoding the discoidin domain receptor 1 (DDR1), a member of a small subfamily of receptor tyrosine kinases, is known to be involved in differentiation, proliferation, and cell adhesion. The extracellular discoidin domain is responsible for the binding of the ligand collagen. As the human homologue, the rat Ddr1 gene consists of 17 exons and is located in the major histocompatibility complex, the RT1 complex in rats. A novel testis-specific Ddr1 transcript of 3.5 kb is described here which is expressed specifically in the postmeiotic germ cells of the rat testis. The exons 1 to 5 are missing in this transcript and the putative protein would lack the discoidin domain and parts of the stalk region. The expression level of both, the full-length 4.3 kb and the novel 3.5 kb Ddr1 transcript, is dependent on the RT1 haplotype. In the RT1(avl) haplotype, carried by DA and LEW.1AV1 rats, the 3.5 kb Ddr1 transcript is completely missing. This might be explained by the lack of four nucleotides GGGC in the RT1(avl) haplotype, which appear to contribute to a SP1 binding site in intron 5 of the Ddr1 gene in the presumed testis-specific alternative promoter region of the 3.5 kb Ddr1 transcript. In addition, two novel exons in the 5' untranslated region of the Ddr1 gene were found that give rise to further alternative Ddr1 transcripts. Interestingly, the 3.5 kb Ddr1 transcript is not only expressed in a cell type-specific manner in postmeiotic germ cells but also controlled by the RT1 haplotype. PMID- 16472942 TI - Ohanin, a novel protein from king cobra venom: its cDNA and genomic organization. AB - Ohanin, from king cobra venom, is a novel protein which induces hypolocomotion and hyperalgesia in mice [Pung, Y.F., Wong, P.T.H., Kumar, P.P., Hodgson W.C., Kini, R.M., 2005. Ohanin, a novel protein from king cobra venom induces hypolocomotion and hyperalgesia in mice. J. Biol. Chem. 280, 13137-13147.]. It is weakly similar to PRY-SPRY domains (B30.2-like domain). Here we report the complete cDNA and genomic organization of ohanin. Interestingly, cDNA sequence does not show significant sequence similarity to any known sequences, including those of B30.2-like domain-containing proteins. Its full-length cDNA sequence of 1558 bp encodes for prepro-ohanin with a propeptide segment at the C-terminal. Ohanin is the first member of a new subfamily of proteins containing B30.2-like domain with short N-terminal segment. We named this subfamily as vespryns. There are two mRNA subtypes differing in their 5'-untranslated regions. Southern hybridization study shows that ohanin is encoded by a single gene. Its genomic sequence is 7086 bp with five exons and four introns, and the two types of mRNAs are generated by alternative splicing of exon 2. Our results indicate that ohanin and vespryns may have evolved from the same ancestral gene as B30.2 domain. PMID- 16472943 TI - Functional analysis on the 5'-flanking region of carp fast skeletal myosin heavy chain genes for their expression at different temperatures. AB - Two types of the fast skeletal myosin heavy chain (MYH) genes were cloned from a genomic DNA library of carp (Cyprinus carpio L.) and named MYH10 and MYH30, which showed the sequence similarity to the MYH cDNAs predominantly expressed in carp acclimated to 10 and 30 degrees C, respectively. The 5'-flanking region of about 3 kbp in size each from MYH10 and MYH30 contained various cis-elements to bind to transcriptional regulatory factors such as MyoD family and myocyte enhancer factor 2 (MEF2) family members. To localize functional regions responsible for the MYH gene expression in a temperature-dependent manner, a series of deletion constructs were prepared from the 5'-flanking region, inserted upstream the luciferase gene in a commercially available plasmid, and injected into the dorsal fast muscle of carp acclimated to 10 and 30 degrees C. The sequence of -1004 to 995 bp with the transcriptional activity in MYH30 was identified as an MEF2 binding site. While the activity given by a sequence of -921 to -824 bp in MYH10 contained only a GATA box, that of the activity of the -1 kbp construct from MYH10 was markedly higher in carp reared at 10 degrees C than fish reared at 30 degrees C. On the other hand, no temperature-dependent expressional regulation was observed for MYH30 even with the full-length construct of -3 kbp. The DNA fragment of -921 to -824 bp in MYH10 and MEF2 binding site in MYH30 interacted with nuclear proteins extracted from carp fast skeletal muscle as revealed by electrophoretic mobility shift assay. The signal intensity of a complex formed between the DNA fragment of MYH10 and nuclear extracts from the 10 degrees C acclimated carp were higher than those with extracts from the 30 degrees C acclimated fish. Although MEF2-binding site in MYH30 could form complex with nuclear extracts from the 30 degrees C-acclimated carp, the same or stronger signals were detected in complex formed with extracts from the 10 degrees C acclimated fish. PMID- 16472944 TI - Organization and structure of hox gene loci in medaka genome and comparison with those of pufferfish and zebrafish genomes. AB - We isolated BAC clones that cover the entire hox gene loci in the medaka fish Oryzias latipes. The BAC clones were characterized by the Southern hybridization with many hox gene probes isolated in our previous study and by PCR using primers designed for selective amplification of respective hox genes. Then, the BAC clones have been subjected to shotgun sequencing. The results revealed the organization of the entire hox gene loci. Forty-six hox genes in total are encoded in seven clusters as follows: 10 hox genes in Aa cluster; 5 in Ab; 9 in Ba; 4 in Bb; 10 in Ca; 6 in Da; and 2 in Db. Together with the information on the hox gene loci registered in the Fugu genome database and in the Danio genome database, the physical maps of three fish genomes were constructed and compared one another. Not only numbers of hox genes but also the distances between the neighboring hox genes are highly similar between medaka and fugu. As for six clusters, Aa, Ab, Ba, Bb, Ca and Da that are commonly present in the three fishes, only few or no differences were found in each cluster. Thus, the hox gene sets should have been well conserved once they had been established in respective species. PMID- 16472945 TI - Genomic organization, characterization, and molecular 3D model of GDE1, a novel mammalian glycerophosphoinositol phosphodiesterase. AB - Glycerophosphodiester phosphodiesterase (GDPD) catalyzes the hydrolysis of deacylated glycerophospholipids to glycerol phosphate and alcohol. A mammalian glycerophosphoinositol phosphodiesterase, GDE1/MIR16, was recently identified as an interacting protein of the regulator of G protein signaling 16 (RGS16) providing a link between phosphoinositide metabolism and G protein signal transduction. To further understand the function and properties of human GDE1, we determined its genomic organization and its biochemical and structural characteristics. GDE1 encodes a 331-residue protein with two hydrophobic domains and contains a GDE domain that shares strong homologies with GDE1-related proteins as well as bacterial GDPDs. The human GDE1 gene is located on chromosome 16p12-p11.2 and contains six exons and five introns. A molecular 3D model, which was built based on the crystal structure of Escherichia coli GDPD (1YDY), provides the first structural information of human GDE1 and suggests a TIM barrel core as typically found in bacterial GDPDs. Furthermore, a model of the putative catalytic motif within the GDE domain was nearly identical to the corresponding domain of GDPD and highlights the individual core residues Glu97, Asp99, and His112, which are crucial to maintaining GDE1 catalytic activity. These studies provide important new insights into understanding the function of GDE1 and GDE1 related proteins. PMID- 16472946 TI - Pulsed-field gel electrophoresis of Campylobacter jejuni sheep abortion isolates. AB - Campylobacter species are a significant cause of sheep abortion in most sheep raising countries. In New Zealand, Campylobacter fetus subsp. fetus is the leading cause of diagnosed sheep abortion and the species C. jejuni and C. coli have also been implicated. To date, strain typing information of C. jejuni sheep abortion isolates is limited. The objective of the present study was to genotype C. jejuni isolates cultured from sheep abortions submitted to diagnostic laboratories in New Zealand during the 2000 breeding season, using pulsed-field gel electrophoresis (PFGE). In this study, C. jejuni isolates were cultured from approximately 10% of farms from which Campylobacter species were isolated from sheep abortions in the year 2000. This equated to 25 C. jejuni isolates from 21 farms. These isolates were obtained from the veterinary diagnostic laboratories and strain typed using the molecular typing technique PFGE. Ten distinct PFGE types were identified amongst the isolates. No particular PFGE type was found most frequently amongst these C. jejuni sheep abortion isolates. However, indistinguishable or similar C. jejuni PFGE types were identified from different aborted foetuses from the same flock, consistent with the role of C. jejuni as an infectious cause of abortion in sheep. These strain types were similar or indistinguishable from C. jejuni sheep abortion isolates obtained in 1999 in a smaller study (Mannering, S.A., Marchant, R.M., Middelberg, A., Perkins, N.R., West, D.M., Fenwick, S.G., 2003. Pulsed-field gel electrophoresis typing of C. fetus subsp. fetus from sheep abortions in the Hawke's Bay region of New Zealand. NZ Vet. J. 51, 33-37). PMID- 16472947 TI - Physiological development of a mixed metabolic and respiratory umbilical cord blood acidemia with advancing gestational age. AB - OBJECTIVE: To investigate respiratory and metabolic components of gestational age dependent umbilical cord blood gas changes. STUDY DESIGN: Cord blood gases were determined in 1336 vigorous singletons with uncomplicated cephalic vaginal delivery at 37-43 weeks. Linear regression analysis and non-parametric statistics were used with a P < 0.05 being significant. RESULTS: Gestational age correlated negatively with arterial pH and HCO3- and positively with pCO2 and base deficit. Venous blood showed corresponding changes except for pCO2, which was independent of gestational age. Arterial pCO2, but not venous, correlated positively with birthweight deviation from the mean. The fractional fetal CO2 production per birthweight unit and the fractional placental CO2 clearance per placental weight unit were not correlated with gestational age. CONCLUSIONS: A mixed respiratory and metabolic cord blood acidemia develops with advancing gestational age. The respiratory component depends on an increased 'CO2 load' from the growing fetus and not on deterioration of placental gas exchange. The etiology of the metabolic component is yet unknown. PMID- 16472948 TI - Association between life stress during pregnancy and infant crying in the first six months postpartum: a prospective longitudinal study. AB - OBJECTIVE: To examine prospectively the relationship between prenatal life stress and infant crying/fussing during the first 6 months of postnatal life, taking into account an array of confounders suggested in the literature. DESIGN: Prospective longitudinal study of a convenient sample, with data points in pregnancy and at about 6 weeks, 3, and 6 months postpartum. METHODS: The study included 86 pregnant women who completed a standardized, validated and widely used questionnaire on negative life changes experienced in the preceding 12 months. Women were grouped by median split on the impact score of negative life changes. Demographic, obstetric and lifestyle variables were obtained from pre- and postnatal interviews and from medical records in order to be taken into account as possible confounders. At all three postnatal data points, mothers kept a validated 5-day 24-h behavior diary to assess durations of infant crying/fussing. RESULTS: Infants of mothers with high scores of negative life changes exhibited more crying/fussing than infants born to mothers with low negative change scores, throughout the first half year postpartum, but particularly at ages 3 and 6 months. These results do not seem to be spurious due to the confounders considered in this report or to recording bias. CONCLUSION: Prenatal life stress is associated with infant crying/fussing in the first half year after delivery. To prevent or reduce infant crying and to foster a well adapted parent-infant relationship, professionals attending expectant mothers should consider their emotional condition. If required, support should be provided already in pregnancy. PMID- 16472949 TI - Evaluation of eye and skin irritation of arginine-derivative surfactants using different in vitro endpoints as alternatives to the in vivo assays. AB - Arginine-derivative surfactants constitute a novel class of surfactants, which can be regarded as an alternative to conventional surfactants. Prior to human exposure, it is necessary to assess their irritation potential. The classical in vivo evaluation of the irritancy potential via the Draize test has been extensively criticized. In that regard, a great number of in vitro alternatives have been developed. Erythrocytes were chosen as the target cells for eye irritation assessment and hemolysis and hemoglobin denaturation were selected as appropriate endpoints. For skin irritancy assessment, the keratinocyte cell line NCTC 2544 was used and different in vitro endpoints were measured: two cytotoxicity assays (NRU and MTT) and the synthesis of the proinflammatory cytokine IL-1alpha. The eye and skin Draize tests were also performed for comparative purposes. The results point out that, according to in vivo and in vitro assays, the new arginine-derivative surfactants have lower eye and skin irritation potential than the synthetic surfactant SDS. Furthermore, in vitro methods were also able to detect differences in irritancy among the new surfactants not noticeable by the Draize tests, indicating that in vitro methods can be more sensitive than the in vivo test, offering the opportunity to detect subtle differences in irritancy. PMID- 16472950 TI - Parsing the ageing Asian woman: symptom results from the China study of midlife women. AB - OBJECTIVES: It has become popular to assert that Asian women in general have low midlife rates of reporting a wide variety of symptoms and that these rates are much lower than those found among women in western countries. Comparable data from mainland China remains sparse, however. This study aimed to uncover the symptom reporting frequency for a general population sample of women in mainland China and to compare these results with rates from similar studies conducted in Japan and North America. METHODS: The author conducted a community-based survey of over 400 Chinese women aged 40-65 years living in a rural village and an urban neighborhood in northern China. The cross-cultural analysis was conducted on the 156 Chinese women within that sample who were between the ages of 45 and 55 years. Their rates of reporting 16 core symptoms in the previous 2 weeks were compared with those of women in the same age range in studies conducted in Japan [Lock M. Encounters with aging: mythologies of menopause in Japan and North America. Berkeley: University of California Press; 1993], Canada [Kaufert P, Lock M, McKinlay S, et al. Menopause research: the Korpilampi workshop. Soc Sci Med 1986;22(11):1285-9], and the U.S. [McKinlay SM, McKinlay JB. Research note: aging in a 'healthy' population. Soc Sci Med 1986;23(5):531-5]. RESULTS: The Chinese women had a low to moderate rate (below 50%) of reporting each of the 16 core symptoms. On most symptoms, the Chinese rates were higher than the Japanese and more similar to the North American. The only exceptions were hot flashes and headaches. CONCLUSIONS: Data from the China Study of Midlife Women showed that women's rates of midlife core symptom reporting in mainland China differ from the extremely low levels found in Japan. It is important to avoid generalizations about East Asian women relative to North American women in this regard. PMID- 16472952 TI - Development of the human fetal cochlear nerve: a morphometric study. PMID- 16472951 TI - Partitioning of parabens between phases of submicron emulsions stabilized with egg lecithin. AB - Partitioning of methyl and propyl parabens (methyl and propyl hydroxybenzoate, paraben M and P) between the major phases in the parenteral submicron emulsions was studied. The investigated emulsions contained 10% or 20% soya-bean oil, 1.2% or 2.4% egg lecithin, 0.18% or 0.36% paraben M and 0.02% or 0.04% paraben P. The aqueous phase was obtained by ultracentrifugation, and subsequently, it was subjected to ultrafiltration, which procedure allowed to distinguish between the fractions of free preservatives (Fw) and incorporated in the liposomal or micellar region (Flm). The fractions present in the oily phase and in the interface were calculated. Depending on the formulation, Fw was 17-31% and 2.3 6.0% for paraben M and P, respectively. The Flm values were in a very narrow range, i.e. 3.0-6.0% for both preservatives. Substantial accumulation, i.e. 38 58% was found in the interface and the partitioning into this region was related to the oil/lecithin ratio rather than to lipophilicity of the preservative. PMID- 16472953 TI - Proposal for internet-based Digital Dental Chart for personal dental identification in forensics. AB - A dental chart is very useful as a standard source of evidence in the personal identification of bodies. However, the kind of dental chart available will often vary as a number of types of odontogram have been developed where the visual representation of dental conditions has relied on hand-drawn representation. We propose the Digital Dental Chart (DDC) as a new style of dental chart, especially for open investigations aimed at establishing the identity of unknown bodies. Each DDC is constructed using actual oral digital images and dental data, and is easy to upload onto an Internet website. The DDC is a more useful forensic resource than the standard types of dental chart in current use as it has several advantages, among which are its ability to carry a large volume of information and reproduce dental conditions clearly and in detail on a cost-effective basis. PMID- 16472955 TI - Lymphotropic nanoparticle enhanced MR imaging (LNMRI) technique for lymph node imaging. AB - Accurate nodal staging is important in the management of any primary malignancy. The presence of nodal metastases has both therapeutic and prognostic implications. Lymphotropic nanoparticles are a new class of MRI contrast agents, which are promising in detecting minimal metastatic nodal disease particularly in normal sized lymph nodes. This paper discusses the technique and interpretation of lymphotropic nanoparticle enhanced MRI (LNMRI) and reviews the various trials evaluating nodal staging with ferumoxtran-10 enhanced MRI. PMID- 16472956 TI - Relationship between HPLC precision and number of significant figures when reporting impurities and when setting specifications. AB - The rounding of an analytical result is a process that should take into account the uncertainty of the result, which is in turn assessed during the validation exercise. Rounding rules are known in physical and analytical chemistry since a long time, but are often not used or misused in pharmaceutical analysis. The paper describes the theoretical background of the most common rules and their application to fix the rounding of results and specifications. The paper makes use of uncertainty values of impurity determination acquired during studies of reproducibility and intermediate precision with regards to 22 impurities of drug substances or drug products. As a general rule, authors propose the use of sound and well-established rounding rules to derive rounding from the results of the validation package. PMID- 16472959 TI - Physical activity counseling in primary care: who has and who should be counseling? AB - OBJECTIVE: To examine the physical activity (PA) counseling literature in primary care in order to identify which intervention provider has been used to date and their relative effectiveness for increasing PA. METHOD: MEDLINE and PsycINFO databases were searched for PA intervention studies in primary care settings. RESULTS: Of the 19 studies, 37% were conducted solely by physicians, 37% by allied health professionals, while 26% were combined-provider interventions. There was a decline in the number of physician-only interventions and a shift towards interventions offered by allied health professionals as adjuncts or alone. Interventions across all provider categories generated some improvements in physical activity behavior, however, it appears that allied health professionals as adjuncts or alone produced the best results in the long-term (>6 months). There was substantial variation in the location and counseling approach employed by allied health professionals. CONCLUSION: We argue for an interdisciplinary model in which physicians recommend PA and provide referrals to allied health professionals such as physical activity counselors. PRACTICE IMPLICATIONS: With physical activity counselors' specialized training and greater time available to the patient, they may provide more intensive and effective counseling required for behavior change and maintenance. PMID- 16472958 TI - Contribution of inflammatory processes to Alzheimer's disease: molecular mechanisms. AB - There is compelling evidence that Alzheimer's disease (AD) amyloid-beta (Abeta) deposition is associated with a local inflammatory response, which is initiated by the activation of microglia and the recruitment of astrocytes. These cells secrete a number of cytokines and neurotoxic products that may contribute to neuronal degeneration and cell death. It has been documented that long-term intake of non-steroidal anti-inflammatory drugs (NSAIDs) decrease the risk for developing AD and delay the onset of the disease. The mechanism behind these NSAIDs is still controversial and several hypotheses have been raised, including changes in the amyloid precursor protein (APP) metabolism, in Abeta aggregation and a decrease in inflammatory mediators. Recently, it was proposed that some NSAIDs might activate the peroxisome proliferator-activated receptor-gamma (PPAR gamma). PPAR-gamma belongs to a family of nuclear receptors that are able to regulate the transcription of pro-inflammatory molecules, such as iNOS. The activation of PPAR-gamma has been recently reported to reduce Abeta levels in cell culture and AD animal models. The implication of PPAR-gamma in the control of Abeta-induced inflammation suggests a new target for AD therapy and emphasize the contribution of neuroinflammatory mechanisms to the pathogenesis of AD. PMID- 16472960 TI - Pictures and text in instructions for medical devices: effects on recall and actual performance. AB - OBJECTIVE: The present study aimed to contribute to the design of effective health education information. Based on cognitive-psychological theory, pictures were expected to improve understanding of two existing textual instructions for using asthma devices (inhaler chamber and peak flow meter). From an analysis of the affordances and constraints of both devices this effect was expected to be stronger with the inhaler chamber than with the peak flow meter. METHODS: To test this, both instructions were systematically illustrated with seven line-drawings visualizing the actions. In two separate randomized controlled trials with in total 99 participants from the general public, the original text-only versions were compared to the text-picture versions of the same instruction. Dependent variables were participants' recall of the instructions and the quality of their performance with the instruction observed from video-recordings. RESULTS: Conform expectations, the results showed significant positive effects of pictures on recall and performance in both instructions, especially with the inhaler chamber. CONCLUSION: Thus, pictures may contribute to a better comprehension and use of medical devices that are inherently less clear. PRACTICE IMPLICATIONS: Health educators may optimize instruction design by careful analysis of the device with instruction and observational testing with potential users. PMID- 16472962 TI - [Combined ilio-inguinal and iliohypogastic block]. PMID- 16472961 TI - Acromegaly due to a somatroph adenoma in a dog. AB - A 10-year-old uncastrated male Dalmatian dog was referred for gait abnormalities consisting of chronic progressive stiffness and rigidity. Other symptoms were polyphagia associated with weight gain, polyuria and polydipsia, excessive panting, and an inspiratory stridor. The owner had noticed progressive thickening of the skin and enlargement of the tongue over the last 3 years. Physical examination revealed thickening of the skin, redundant skin folds, and enlargement of the tongue. The only remarkable abnormalities found on routine laboratory examination were mild anaemia and an increased serum fructosamine concentration. Circulating concentrations of total thyroxine, free thyroxine, and cTSH, and the results of an ACTH stimulation test were all within reference ranges. The basal serum growth hormone (GH) concentration was markedly elevated (23microg/l) and did not decrease during a glucose tolerance test or after somatostatin administration. The serum insulin-like growth factor-1 concentration was also markedly elevated (1254microg/l). Basal serum insulin concentration was high (95mU/l) and insulin concentrations increased considerably after glucose loading, consistent with insulin resistance. Abdominal ultrasonography showed no abnormalities. Survey radiographs of the vertebral column showed severe spondylosis deformans extending from the cervical to the lumbosacral spine. CT scanning of the skull showed an enlarged pituitary gland with normal enhancement pattern. On post-mortem examination, the entire vertebral column appeared as a single and inflexible structure due to the presence of multiple fused osteophytes. The pituitary gland contained an acidophilic adenoma that immunostained positively for GH (and negatively for ACTH and alpha-MSH). In conclusion, this Dalmatian dog with acromegaly and insulin resistance represents the first case of GH hypersecretion proven to be due to a somatotroph adenoma. PMID- 16472963 TI - Protective effect of L-propionylcarnitine in chronic cyclosporine-a induced nephrotoxicity. AB - Cyclosporine (CyA) is an immunosuppressive agent used after solid organ transplantation, but its clinical use is limited by side effects, the most important of which is nephrotoxicity. In a previous work we demonstrated that L propionylcarnitine (L-PC), a propionyl ester of L-carnitine, is able to prevent CyA-induced acute nephrotoxicity reducing lipid peroxidation in the isolated and perfused rat kidney. CyA administration was associated with a dose dependent increase in renovascular resistance prevented by a pretreatment with L-PC. The aim of the present study was to confirm L-PC protective effect, previously described in vitro, in an in vivo rat model. Chronic nephrotoxicity study was carried out for 28 days. L-PC was administered (i.p. 25 mg/kg b.w.) since the first day, while CyA treatment was performed for the last 21 days (by oral administration 25 mg/kg b.w.). We demonstrate that L-PC was able to significantly lower blood pressure in CyA treated animals and to prevent CyA induced decrease in creatinine clearance. Moreover renal tissue analysis revealed that L-PC was able to reduce lipid hydroperoxide content and morphological abnormalities associated to chronic CyA administration. In conclusion our study demonstrated for the first time in vivo that L-PC protects against functional and tissue damage associated to chronic CyA administration. PMID- 16472964 TI - Differential induction of apoptosis by type A and B trichothecenes in Jurkat T lymphocytes. AB - Several studies have shown that the mycotoxins T-2 toxin, diacetoxyscirpenol (DAS), deoxynivalenol (DON) and nivalenol (NIV) affect lymphocyte functioning. However, the molecular mechanisms underlying the immunomodulatory effects of these trichothecenes are not defined yet. In this study, the potency of the type A trichothecenes T-2 toxin and DAS, and the type B trichothecenes DON (and its metabolite de-epoxy-deoxynivalenol; DOM-1) and NIV to reduce mitochondrial activity and to induce apoptosis of Jurkat T cells (human T lymphocytes) were examined. T-2 toxin and DAS are much more cytotoxic at low concentrations than DON and NIV as shown by the AlamarBlue cytotoxicity assay. In addition, the mechanism whereby DON and NIV induced cytotoxicity is mainly via apoptosis as we observed phosphatidylserine externalization, mitochondrial release of cytochrome c, procaspase-3 degradation and Bcl-2 degradation. In contrast, type A trichothecenes reduce the mitochondrial activity at approximately 1000-fold lower concentrations than the type B trichothecenes, resulting in necrosis. These data suggest that the mechanisms resulting in cytotoxic effects are different for type A and type B trichothecenes. PMID- 16472965 TI - Cytotoxic effect of As(III) in Caco-2 cells and evaluation of its human intestinal permeability. AB - Inorganic arsenic has been classified as a carcinogen for humans (Group I). However, its transit across the human intestinal epithelium has not been characterized. Using Caco-2 cells, the thiol-redox balance and apparent permeability coefficients (P(app)) for As(III) in the apical to basolateral (AP BL) and basolateral to apical (BL-AP) direction were evaluated. After As(III) exposure, GSH-induced synthesis was observed, increasing the GSH/GSSG ratio by elevating the As(III) concentration. The AP-BL permeabilities decreased as the As(III) concentrations increased, indicating the existence of a mediated transport mechanism. The (BL-AP)/(AP-BL) permeability ratios were higher than unity, suggesting the existence of a secretion process. PMID- 16472966 TI - Effect of pyridoxylidene aminoguanidine on human diploid cells B-HEF-2: in vitro cytotoxicity test and cytogenetic analysis. AB - BACKGROUND: Pyridoxylidene aminoguanidine is an appropriate inhibitor of protein glycation, respectively formation of advanced glycation products, which are connected with mechanism of pathogenesis in chronic diabetic complications. Moreover, it was found that in comparison with aminoguanidine, pyridoxylidene aminoguanidine does not influence the level of vitamin B6 in liver and kidneys in vivo. The aim of this study was to test cytotoxic effect of pyridoxylidene aminoguanidine in vitro, in regard to its potential use as inhibitor of advance protein glycation in diabetic patients. METHODS: The potential genotoxic activity of pyridoxylidene aminoguanidine in vitro was assessed by the micronucleus test and the karyological analysis. The direct contact method using diploid human cell line B-HEF-2 was performed to evaluate cytotoxicity. The concentrations of 5 x 10(-3), 2.5 x 10(-3) and 1 x 10(-3) ml/l were used in all tests. RESULTS: Microscopic analysis did not proved any changes in morphology of exposed fibroblasts. The inhibitive effect of pyridoxylidene aminoguanidine was increased with rising concentration. The proliferative activity of exposed cells to concentrations of 1 x 10(-3), 2.5 x 10(-3), 5 x 10(-3) mol/l was inhibited approximately by 30%, 60% and 80%, respectively. The frequency of micronuclei and rate of numerical or structural aberrations was not increased. CONCLUSION: Obtained results confirmed that pyridoxylidene aminoguanidine in selected concentrations has an inhibitive effect on the proliferation activity of exposed cells, but did not develop any cytotoxic effect on B-HEF-2 cells. PMID- 16472967 TI - Study of PMS777, a new type of acetylcholinesterase inhibitor, in human HepG2 cells. Comparison with tacrine and galanthamine on oxidative stress and mitochondrial impairment. AB - Acetylcholinesterase inhibitors are commonly used as cognitive enhancers for dementia in aged people. Among them, tacrine (THA) but not galanthamine, was shown to exhibit hepatotoxicity which reduces its clinical use. PMS777, both a PAF antagonist and a new potent acetylcholinesterase inhibitor was recently demonstrated to reverse scopolamine-induced amnesia in mice without toxicity. In the present study, the effects of THA, galanthamine and PMS777 were compared in HepG2 cells on the oxidative parameters involved in the reported hepatotoxicity of THA. THA (> or = 10 microM) induced an oxidative stress as shown by elevated ROS and MDA production and by a decrease in GSH level. Moreover, mitochondrial membrane potential and redox status were decreased. At low concentrations (< or =10 microM), there was no significant disturbance. None of the oxidative stress markers was affected by PMS777 up to the maximum concentration tested and it is suggested that PMS777 is not cytotoxic for HepG2 cells. Galanthamine was also without cytotoxicity. Our results suggest that the toxic effect of THA above 10 microM may be caused by drug-induced mitochondrial energization impairment and destabilisation of membrane phospholipids associated with an oxidative stress. In contrast by preventing these dysfunctions, PMS777 could be safer than THA. PMID- 16472968 TI - Lack of effect on rat testicular organogenesis after in utero exposure to 3 monochloropropane-1,2-diol (3-MCPD). AB - 3-Monochloropropane-1,2-diol (3-MCPD) is a food-born contaminant known to display toxic effects on male reproduction, producing infertility in rats and humans. Using the rat as a model, we investigated whether or not testicular organogenesis, which, in the rat species, occurs during the second half of gestation, was at particular risk regarding 3-MCPD toxicity. Pregnant rats were given daily doses of 5, 10 or 25 mg/kg BW of 3-MCPD from days 11.5-18.5 postcoitum (dpc). On 19.5 dpc, testes were removed from fetuses for histological examination and testosterone analysis. Eight genes were selected among the differentiation markers of testicular cell lineages, and their expression was studied by RT-PCR. The levels of 3-MCPD and its main metabolite, beta chlorolactic acid, were assayed in fetal tissues and dam plasma. Our results show a statistically significant decrease in the mean body weight gain of pregnant rats treated with 10 and 25 mg/kg BW of 3-MCPD. Fetal testes exposed to 3-MCPD exhibited normal histology and produced testosterone at levels that were similar to controls. In addition, 3-MCPD did not alter gene expression in the fetal testes. This lack of effect occurred under conditions where 3-MCPD and beta chlorolactic acid were found to readily cross the placental barrier and diffuse throughout the fetal tissues. Our findings indicate that 3-MCPD has minimal effect on rat testicular organogenesis. PMID- 16472970 TI - Reproductive outcomes following environmental exposure to DDT. AB - We used official statistics of births and stillbirths in 1945-1954 to evaluate reproductive outcomes in the general population following use of DDT during a 1946-1950 anti-malarial campaign in the Italian region of Sardinia. Due to the disruption of registration systems in the World War II years, data in the pre-DDT years were available only for 1945-1946. Such a short period of observation, and social conditions in the war and post-war years, do not allow exclusion of adverse effects of DDT on birth rate; however, we did not observe an effect. The stillbirth rate, infant mortality rate, and male/female ratio in newborns were apparently unaffected following widespread but focused use of DDT in Sardinia, Italy. PMID- 16472969 TI - Sub-chronic administration of diphenyl diselenide potentiates cadmium-induced testicular damage in mice. AB - Sub-chronic cadmium (Cd) exposure causes testicular damage in mice. The mode of action may involve oxidative stress and especially lipid peroxidation. The present study has monitored the pathogenesis of testicular damage during sub chronic Cd exposure and has evaluated the potential protective effect of antioxidant therapy with diphenyl diselenide (PhSe)(2). Male mice were dosed with 2.5 mg/kg CdCl(2) (2.5 mg/kg) with or without (PhSe)(2) (5 micromol/kg) at 30 min post-exposure using a model of five weekly subcutaneous injections. Histological evaluation of the testis was performed across a 4 week test period. Animals exposed to CdCl(2) and CdCl(2) plus (PhSe)(2) displayed a reduction in body weight gain and testicular weight. Progressive damage and histolopathological changes in the testis were not remedied with, but rather were potentiated by, (PhSe)(2) therapy. We conclude that (PhSe)(2) enhances testicular injury in an animal model for sub-chronic Cd exposure mice. PMID- 16472971 TI - Prenatal cocaine dampened behavioral responses to methylphenidate in male and female adolescent rats. AB - Clinical and animal data point toward deficits in attention and arousal after prenatal cocaine exposure. Since methylphenidate (MPD) is widely used to treat attention disorders, we wanted to determine whether prenatal cocaine (PC) exposure affects the behavioral response to MPD in young rats of both sexes. Pregnant dams received 60 mg/kg of cocaine or vehicle from gestational days 8-22 by intragastric intubations. After delivery, litters were culled to 10 (5 males, 5 females) and fostered. On a single day between PND 41-44 locomotion was recorded in a Plexiglas box within an Accuscan activity monitor after receiving a single injection of 10 mg/kg intraperitoneally of MPD or saline. Rats were also videotaped for analysis of stereotyped behavior. Results showed that MPD administration enhanced locomotion compared to saline injected groups. PC exposure in male rats did not have any effect on the locomotor response to MPD compared to prenatal controls. However, PC-exposed males showed a lower amount of time spent in low intensity stereotypy compared to prenatal control males and both groups of females that received MPD. PC exposure in female rats that received MPD dampened the locomotor response compared to prenatal control females that also received MPD. In conclusion PC exposure dampens the behavioral response to MPD differentially in males and females with an apparent selectivity of locomotion in females and stereotyped behavior in males. PMID- 16472972 TI - Symptoms of extraesophageal reflux in a community-dwelling sample. AB - OBJECTIVES: Extraesophageal reflux disease (EERD) is linked with serious and varied health concerns. The authors' goal was to determine the prevalence of EERD and the effect on health-related quality of life in adults within a large community-based sample. STUDY DESIGN: Survey study conducted in a community setting. METHODS: In the Madison, Wisconsin area, 1845 adult subjects were surveyed. Main outcome measures were frequently of GERD and laryngeal symptoms, the Medical Outcomes Trust Short Form 12 (SF-12), Voice Handicap Index (VHI), and Quality of Life in Reflux and Dyspepsia (QOLRAD) questionnaires. RESULTS: More than 66% of subjects reported either GERD or laryngeal symptoms, and 26% reported both GERD and laryngeal symptoms (which reflect symptoms commonly reported for EERD). In addition, 39% of our sample took medicine for heartburn. Forty-four percent of subjects with both GERD and laryngeal symptoms reported occasional breathing difficulties, and 38% of these subjects reported a voice disorder. Significant reductions in perceived quality of general health, digestive health, and voice-related quality of life were found in subjects with both GERD and laryngeal symptoms, in comparison with subjects manifesting each symptom group alone, or in subjects with no symptoms. CONCLUSIONS: The presence of GERD and laryngeal symptoms, and their concurrence, was identified in a substantial number of subjects. The GERD and laryngeal symptoms surveyed represent those most commonly reported for EERD. Because EERD has been linked with tissue damage and reductions in health-related quality of life, it is important to identify these persons and provide treatment before progression of the condition. PMID- 16472973 TI - Transoral robotic surgery (TORS): glottic microsurgery in a canine model. AB - OBJECTIVES/HYPOTHESIS: We hypothesize that bimanual, three-dimensional robotic surgery will prove valuable for glottic microsurgical procedures. STUDY DESIGN: To test this hypothesis, we developed and optimized a canine model for glottic microsurgery using a commercially available surgical robot. Methods Using a da Vinci Surgical Robot (Intuitive Surgical, Inc., Sunnyvale, CA), glottic microsurgery was performed with a hydrodissection technique in a canine model. The experiments were performed on two orotracheal intubated mongrel dogs under general anesthesia in the supine position on a standard operating room table. A videoscope and two, 360-degree rotating, 5- and 8-mm, wrested-end effector instruments were introduced transorally with three robotic arms. The surgeon performed the actual procedures while positioned at a robotic system console that was located across the operating room suite. The procedure was performed in duplicate and was documented with still and video photography. RESULTS: Glottic microsurgery was successfully performed using the da Vinci Surgical Robot, with both 5- and 8-mm instrumentation. The smaller, 5-mm instruments afforded greater visualization of the operative site and increased maneuverability, which resulted in reduced operative time. The robotic system provided excellent visualization as well as controlled and delicate microdissection at the vocal cord level. CONCLUSIONS: Robotic laryngeal microsurgery is technically feasible in the canine model and warrants consideration for evaluation in controlled human trials. PMID- 16472974 TI - Are real-time displays of benefit in the singing studio? An exploratory study. AB - This article reports on an exploratory research project to evaluate the usefulness or otherwise of real-time visual feedback in the singing studio. The primary purpose of the work was not to optimize the technology for this application, but to work alongside teachers and students to study the impact of real-time visual feedback technology use on the students' learning experiences. An action research methodology was used to explore the benefit of real-time displays over an extended period. The experimental phase of the work was guided by a Liaison Panel of teachers and academics in the areas of singing, pedagogy, voice science, speech therapy, and linguistic science. Qualitative data were collected from eight students working with two professional singing teachers. The teachers and students acted as co-researchers under the action research paradigm. Teachers and students alike kept journals of their teaching and learning experiences. Singing lessons were observed regularly by the research team, coded for teacher and student behaviors, and all co-researchers were interviewed at the mid- and endpoint of the project. The use of technology had a positive impact on the learning process, and this is evidenced through case study data. PMID- 16472975 TI - Dietary docosahexaenoic acid-induced generation of liver lipid peroxides is not suppressed further by elevated levels of glutathione in ODS rats. AB - OBJECTIVES: We examined the effects of ascorbic acid (AsA) and glutathione (GSH; experiment 1) and of GSH in acetaminophen-fed rats (experiment 2) on dietary docosahexaenoic acid (DHA)-induced tissue lipid peroxidation. METHODS: In experiment 1, AsA-requiring Osteogenic Disorder Shionogi/Shi-od/od (ODS) rats were fed soybean protein diets containing DHA (10.0% total energy) and AsA at 50 (low) or 300 (normal) mg/kg without (low) or with (normal) methionine at 2 g/kg for 32 d. In experiment 2, ODS rats were fed diets containing DHA (7.8% total energy) and acetaminophen (4 g/kg) with different levels of dietary methionine (low, moderate, high, and excessive at 0, 3, 6, and 9 g/kg, respectively) for 30 d. Tissue lipid peroxides and antioxidant levels were determined. RESULTS: In experiment 1, liver lipid peroxide levels in the low-AsA group were lower than those in the normal-AsA group, but kidney and testis lipid peroxide levels in the low-AsA group were higher than those in the normal-AsA group. Dietary methionine tended to decrease tissue lipid peroxide levels but did not decrease vitamin E (VE) consumption. In experiment 2, a high level of methionine (6 g/kg) decreased liver lipid peroxide levels and VE consumption. However, generation of tissue lipid peroxides and VE consumption were not decreased further by a higher dose of methionine (9 g/kg). CONCLUSIONS: Higher than normal levels of dietary methionine are not necessarily associated with decreased dietary DHA-induced generation of tissue lipid peroxides and VE consumption except that the GSH requirement is increased in a condition such as acetaminophen feeding. PMID- 16472977 TI - Citrus juice modulates bone strength in male senescent rat model of osteoporosis. AB - OBJECTIVE: An experiment evaluated the effect of citrus juice on enhancing serum antioxidant status and on osteoporosis prevention in orchidectomized rats. METHODS: Thirty-six 1-y-old male rats were randomized to two groups: a sham control group (n = 9) and an orchidectomized group (n = 27). The orchidectomized group was divided into three groups of nine and assigned to one of the following treatments: orchidectomy, orchidectomy plus orange juice, and orchidectomy plus grapefruit juice. Sixty days after initiation of the study, all rats were killed, blood was collected, and serum was harvested for total antioxidant status and indices of bone formation and resorption. Femoral density and biomechanical properties were monitored. RESULTS: Orchidectomy decreased (P < 0.05) total antioxidant capacity, femoral density, and biomechanical properties and increased (P < 0.05) alkaline phosphatase, acid phosphatase, and urinary excretion of hydroxyproline compared with the sham-control group. In contrast to orchidectomy, orchidectomy plus orange juice and orchidectomy plus grapefruit juice reversed (P < 0.05) orchidectomy-induced antioxidant suppression, decreased (P < 0.05) alkaline phosphatase and acid phosphatase activities, moderately restored (P = 0.07) femoral density, increased (P < 0.05) femoral strength, significantly delayed time-induced femoral fracture, and decreased (P < 0.05) urinary excretion of hydroxyproline. CONCLUSION: The present study supports the supposition in that drinking citrus juice positively affects serum antioxidant status and bone strength. PMID- 16472976 TI - Response of muscle protein and glutamine kinetics to branched-chain-enriched amino acids in intensive care patients after radical cancer surgery. AB - OBJECTIVE: Patients with cancer are characterized by decreased muscle protein synthesis and glutamine availability that contribute to an impaired immune response. These abnormalities worsen after surgical stress. We tested the hypothesis that pharmacologic doses of branched-chain amino acids would improve the early metabolic response after major cancer surgery. METHODS: By using a crossover experimental design, we compared the metabolic effects of isonitrogenous solutions of balanced and branched-chain-enriched amino acid mixtures infused at the rate of 82 mg x h(-1) x kg(-1) for 3 h in patients with colorectal or cervical cancer on the first and second days after radical surgery combined with intraoperative radiation therapy. The ratios of leucine to total amino acid (grams) in the two mixtures were 0.09 and 0.22, respectively. Muscle protein and glutamine kinetics were determined by using stable isotope of amino acids and the leg arteriovenous balance technique. Glucose and insulin were continuously infused throughout the 2-d study to maintain near euglycemia. RESULTS: Rates of muscle protein synthesis and degradation were not significantly affected by the balanced amino acid infusion. In contrast, the isonitrogenous, branched-chain-enriched amino acid mixture accelerated muscle protein turnover by stimulating (P or=500 microM) of alphaTOC, suggesting an inhibition on nuclear factor kappaB pathway for activation of IL-2 expression. PPARgamma mRNA level in activated splenocytes was upregulated by 1 mM of alphaTOC. PPARgamma mRNA level in unstimulated splenocytes was upregulated by alphaTOC in a dose-dependent manner, suggesting that alphaTOC might enhance the PPARgamma signaling pathway. High-dose alphaTOC induced apoptosis of splenocytes and inhibited phytohemagglutinin-stimulated T-cell proliferation. Conversely, the proliferative response of splenocytes was enhanced by 5 microM of alphaTOC. Low-dose (50 microM) alphaTOC increased IL-2 expression, which may have been due to the absence of downregulation of PPARgamma and IkappaBalpha on the IL-2 gene. CONCLUSION: The results indicated that low and high doses of alphaTOC exert opposite effects on IL-2, possibly through modulation of PPARgamma, IkappaBalpha, and apoptosis pathways. The present findings support our previous observation of opposite effects of low- and high-dose vitamin E on survival of MRL/lpr mice. PMID- 16472985 TI - Prevention of bacterial translocation using glutamine: a new strategy of investigation. AB - OBJECTIVE: This study investigated the role of glutamine (Gln) on bacterial translocation in an intestinal obstruction model by using Escherichia coli labeled with technetium 99m (99mTc-E. coli). METHODS: Intestinal obstruction was performed by a single ligature of the terminal ileum in rats. Animals in the control group (group 1) were sham operated (not obstructed). Experimental group 2 had intestinal obstruction. Groups 1 and 2 were not treated with Gln. Groups 3 and 4 were treated with Gln for 7 d before surgery with 250 and 500 mg x kg(-1) x d(-1), respectively. A suspension containing 100 million colony-forming units/mL of (99m)Tc-E. coli was injected into the lumen of the ileum. Twenty-four hours after surgery, blood, mesenteric lymph nodes, livers, spleens, and lungs were collected for determination of radioactivity. The Mann-Whitney U test was performed for statistical analysis. P 20 years ago. What began as a curiosity in this model organism has expanded into almost every area of biology; there are now 326 confirmed miRNA genes in humans and the total is predicted to reach 1000. Each miRNA has the potential to regulate hundreds of mRNAs; therefore, there are likely to be few biological pathways not impacted by miRNA regulation. Recent evidence has suggested that miRNAs might be viable therapeutic targets for a wide range of diseases, including cancer. A recent article by Stoffel and colleagues has demonstrated remarkably effective inhibition of miRNAs in vivo, thus providing an entry point into the promising new arena of miRNA therapeutics. PMID- 16473044 TI - Lymphadenitis as a major element of disease in the guinea pig model of tuberculosis. AB - Guinea pigs infected by low dose aerosol with the H37Rv strain of Mycobacterium tuberculosis rapidly developed granulomatous lesions in the pulmonary parenchyma and within the intra-thoracic hilar lymph node cluster. Lung lesions showed no predilection for specific lobes and were perivascular, peribronchial and peribronchiolar throughout the subpleural, hilar and pulmonary parenchyma. Marked hilar lymph node enlargement was due to coalescing foci of subcapsular, paracortical and medullary granulomatous inflammation that progressed to necrosis that effaced normal lymph node architecture. Lymph node lesions became severe and progressed more rapidly than pulmonary lesions. Immunization with BCG 6 weeks prior to infection significantly reduced the lung and lymph node lesion burden as well as the progression to necrosis in both tissues. Lymph node inflammation in BCG immunized animals partially resolved and was replaced by fibroblasts and fibrous connective tissue while lesions from non-immunized animals continued to progress to necrosis. We discuss here the observation that the distribution and progression of lung and lymph node lesions in the guinea pig aerosol model of tuberculosis have considerable similarity to the naturally occurring disease in children. PMID- 16473046 TI - "Erratic" complex partial status epilepticus as a presenting feature of MELAS. AB - Patients with the syndrome of mitochondrial encephalopathy, lactic acidosis, and strokelike episodes (MELAS) may rarely present with complex partial status epilepticus (CPSE) at clinical onset. We report on a 38-year-old woman with MELAS presenting with multifocal CPSE and periodic lateralized epileptiform discharges (PLEDs) on EEG during her first strokelike episode. CT scan documented a right temporo-parieto-occipital strokelike lesion. EEG showed prolonged seizure discharges with alternating focus over the temporo-occipital and frontotemporal regions of the right hemisphere; moreover, right frontotemporal PLEDs were evident when the seizure activity was localized in the temporo-occipital region. The electroclinical status and CT findings normalized gradually on carbamazepine therapy. The four other patients with MELAS described in the literature as presenting with CPSE showed unifocal epileptic discharges on EEG. We report for the first time a case in which multifocal CPSE is the presenting feature of MELAS. Our findings document the multifocality of neuronal hyperexcitability in the context of the cerebral strokelike lesion in this syndrome. PMID- 16473047 TI - Haemolymph as compartment for efficient and non-destructive determination of P glycoprotein (Pgp) mediated MXR activity in bivalves. AB - Measurement of the modulation of accumulation rate of model P-glycoprotein (Pgp) substrates has been a well established methodology for determination of the presence and activity of the multixenobiotic resistance (MXR) defence mechanism in aquatic invertebrates. Most studies have been focused on the gill tissue of various bivalves as a primary compartment for this type of measurements. In this study, we evaluated the potential of measuring the accumulation rate of a fluorescent model Pgp substrate rhodamine B (RB) in haemolymph, plasma and haemocytes of the freshwater painter's mussel (Unio pictorum) as additional potentially useful compartments. The obtained results demonstrated several important advantages of the determination of Pgp mediated MXR transport activity in haemolymph over determinations in gill tissue. The overall MXR response correlated well with the level of Pgp activity simultaneously determined in gills. The method is more sensitive, the procedure is easier and less laborious, and repeated use of same individuals is possible. Finally--the approach is non destructive, offering a potentially powerful biomarker and research tool for studies directed to the evaluation of ecotoxicological importance of MXR defence and the presence of MXR inhibitors in the environment. PMID- 16473045 TI - Echocardiographic visualization of laceration of atrial septum during balloon sizing of atrial septal defect. AB - Transcatheter closure of atrial septal defect (ASD) has become an accepted alternative to surgery. A number of complications associated with ASD device closure have been recognized but most are rare or minor in severity. We report a rare complication of atrial septal laceration during transcatheter closure of secundum ASD. We discuss the diagnostic confusion, which resulted in the decision for surgical correction. PMID- 16473048 TI - Resveratrol decreases calcium sensitivity of vascular smooth muscle and enhances cytosolic calcium increase in endothelium. AB - Resveratrol causes endothelium dependent and independent relaxation of vascular smooth muscle. This study investigated the mechanisms behind the effect of resveratrol on vascular tone. Resveratrol (0.1 mM) inhibited KCl-stimulated contractions in endothelium-denuded rat aorta and this inhibition was not reversed by tetraethylammonium (TEA) (5 mM), glyburide (3 microM), ouabain (0.1 mM), thapsigargin (1 microM), or indomethacin (10 microM). KCl (90 mM) increased the intracellular free calcium concentration ([Ca2+]i) in the isolated smooth muscle cells from the rat aorta and resveratrol (0.1 mM) did not inhibit the KCl stimulated [Ca2+]i increase. The CaCl2 (0.1-100 microM) stimulated contractions were inhibited by resveratrol (0.1 mM) in the Triton X-100 skinned smooth muscle of the aorta. In heart valve endothelium, resveratrol (0.1 mM) augmented the acetylcholine (10 microM) stimulated [Ca2+]i increase. Resveratrol-induced augmentation of the acetylcholine-stimulated [Ca2+]i elevation was reversed by glyburide (3 microM), but not by TEA (5 mM). The present study indicated that resveratrol affected vascular smooth muscle and endothelium in different ways. Resveratrol decreased the Ca2+ sensitivity but did not affect the KCl-stimulated [Ca2+]i increase in the vascular smooth muscle. In the endothelial cells, resveratrol enhanced the agonist-stimulated [Ca2+]i increase that might trigger nitric oxide synthesis from endothelial cells. PMID- 16473049 TI - Listeria monocytogenes internalins are highly diverse and evolved by recombination and positive selection. AB - To probe the evolution of internalins with confirmed or suspected roles in Listeria monocytogenes virulence we sequenced the full inlB, inlC2, inlC, inlD, inlE, inlF, inlG, and inlH ORFs from 40 L. monocytogenes isolated from human (n=10) and animal (n=10) clinical cases, foods (n=10), and the natural environment (n=10). inlB and inlE were present in all isolates, representing 26 and 20 alleles, respectively. inlC was found in all lineage I and II isolates and represented 21 alleles. inlC2 and inlD represented 22 and 24 alleles, respectively, and were found in all L. monocytogenes isolates, with the exception of three lineage II isolates, which carried inlH, an apparent fusion of the 5' end of inlC2 with the 3' end of inlD. inlF and inlG were absent from lineage I isolates and represented 16 and 11 alleles, respectively. Average pairwise nucleotide differences per site (pi) ranged from 0.00849 (inlF) to 0.07020 (inlE). Phylogenetic trees generally showed clustering of internalin genes into two major evolutionary lineages consistent with lineages I and II previously assigned by ribotyping. In addition to detection of recombination events within each internalin gene, inlB, inlC, inlC2, and inlF showed significant evidence for positive selection (i.e., selection for an advantageous mutant allele). Overall, our data indicated that (i) internalin genes are highly diverse, (ii) internalin gene sequences cluster consistent with the phylogenetic lineages of L. monocytogenes, (iii) both intragenic recombination and positive selection have contributed to the evolution of L. monocytogenes internalins, and (iv) L. monocytogenes internalins show distinct evolutionary histories. PMID- 16473051 TI - Determination of helicidum and its metabolites in dog plasma by LC/UV/MS/MS and its application to pharmacokinetic studies. AB - A simple, rapid and reliable method was developed for the identification and quantification of helicidum and its metabolites in beagle dog plasma by liquid chromatography/ultra-violet/electrospray ionization-ion trap mass spectrometry (LC/UV/ESI-ITMS). Two metabolites were identified by MS: formylphenyl-O-beta-d pyranosyl alloside (I) and hydroxylmethylphenyl-O-beta-d-pyranosyl alloside (II). UV was used for concentration determination with the wavelength of 270 nm. Liquid liquid extraction was used and the extraction recovery exceeded 90%. Kromacil C(18) column (5 microm, 4.6mm i.d. x 250 mm) was used as the analytical column. Linear detection responses were obtained for helicidum concentration ranging from 1.76 x 10(-4) to 70.4 x 10(-4) micromol/mL (0.050-2.00 microg/mL). The precision and accuracy data, based on intra- and inter-day variations over 3 days, were less than 5%. The limit of determination and quantitation (LOD, LOQ) for helicidum was 0.010 and 0.030 microg/mL, respectively. Pharmacokinetic data of helicidum and the two metabolites were obtained with this method after administration of intravenous injection and a single oral dose of tablets to six beagle dogs, respectively. PMID- 16473050 TI - Regulation of mesenchymal stem cell adhesion and orientation in 3D collagen scaffold by electrical stimulus. AB - Cell adhesion and orientation are important for both natural and engineered tissues to fully achieve physiologic functions. Based on diverse cellular responses induced by electrical stimulus on 2D substrate, we applied non-invasive electrical stimulus to regulate cell adhesion and orientation of bone marrow derived mesenchymal stem cells (MSCs) and fibroblasts in a reconstituted 3D collagen-based scaffold. While fibroblasts were induced to reorient perpendicularly in response to direct current electrical stimulus, rat MSCs showed only slight changes in cell reorientation. Multiphoton microscopy revealed that rat MSCs exhibited much stronger 3D adhesion, which appears to resist cell reorientation. Only in response to a large electrical stimulus (e.g., 10 V/cm), collagen fibers around rat MSCs became disconnected and loosely reorganized. In contrast, the collagen fibers surrounding the fibroblasts were entangled in a random network and became preferentially aligned in the direction of the electrical stimulus. When incubated with integrin antibodies, both fibroblasts and rat MSCs failed to respond to electrical stimulus, providing evidence that integrin-dependent molecular mechanisms are involved in 3D cell adhesion and orientation. Elucidation of physical regulation of 3D cell adhesion and orientation may offer a novel approach in controlling cell growth and differentiation and could be useful for stem cell-based therapeutic application and engineering tissue constructs. PMID- 16473052 TI - Assessment of population structure by single nucleotide polymorphisms (SNPs) in goat breeds. AB - Single nucleotide polymorphisms (SNPs) may be used in biodiversity studies and commercial tasks like traceability, paternity testing and selection for suitable genotypes. Twenty-seven SNPs were characterized and genotyped on 250 individuals belonging to eight Italian goat breeds. Multilocus genotype data were used to infer population structure and assign individuals to populations. To estimate the number of groups (K) to test in population structure analysis we used likelihood values and variance of the bootstrap samples, deriving optimal K from a drop in the likelihood and a rise in the variance plots against K. PMID- 16473053 TI - The natural history of Cri du Chat Syndrome. A report from the Italian Register. AB - The aim of this report is to provide an update on the natural history of the Cri du Chat Syndrome by means of the Italian Register (I.R.). Two hundred twenty patients were diagnosed by standard cytogenetic methods and 112 of these were also characterised by molecular-cytogenetic investigation (FISH). FISH analysis showed interstitial deletions, short terminal deletions and other rare rearrangements not previously correctly diagnosed by standard cytogenetics. The diagnosis was made in the first month of life in 42% and within first year in 82% of cases. The remaining 18% were diagnosed at an age ranging from 13 months to 47 years. At the last follow-up, patient age ranged from 8 months to 61 years. Mortality, already low, has decreased over time as it is lower between 1984-2002 compared to 1965-1983. Mortality was higher in patients with unbalanced translocations resulting in 5p deletions. Our data confirm that the cat-like cry and peculiar timbre of voice are the most typical signs of the syndrome, not only at birth but also later and these are the only signs which might suggest the diagnosis in patients with small deletions and mild clinical picture. A cytogenetic and clinical variability must be underlined. Cardiac, cerebral, renal and gastrointestinal malformations were more frequent in the patients with unbalanced translocations resulting in 5p deletions. Sucking and feeding difficulties and respiratory infections are frequent in the first months or years of life. Intubation difficulties linked to larynx anomalies must be considered. Psychomotor development is delayed in all patients but there is a variability related to deletion size and type as well as other genetic and environmental factors. However, the results showed an improvement in the acquisition of the development skills and progress in social introduction which should encourage caregivers and parents to work together in carrying out the rehabilitative and educational interventions. PMID- 16473054 TI - Inhibitors of cyclo-oxygenase-2 and secretory phospholipase A2 preserve bone architecture following ovariectomy in adult rats. AB - Epidemiological evidence and in vitro data suggest that COX-2 is a key regulator of accelerated remodeling. Accelerated states of osteoblast and osteoclast activity are regulated by prostaglandins in vitro, but experimental evidence for specific roles of cyclooxygenase-2 (COX-2) and secretory phospholipase A2 (sPLA2) in activated states of remodeling in vivo is lacking. The aim of this study was to determine the effect of specific inhibitors of sPLA2-IIa and COX-2 on bone remodeling activated by estrogen deficiency in adult female rats. One hundred and twenty-four adult female Wistar rats were ovariectomized (OVX) or sham-operated. Rats commenced treatment 14 days after surgery with either vehicle, a COX-2 inhibitor (DFU at 0.02 mg/kg/day and 2.0 mg/kg/day) or a sPLA2-group-IIa inhibitor (KH064 at 0.4 mg/kg/day and 4.0 mg/kg/day). Treatment continued daily until rats were sacrificed at 70 days or 98 days post-OVX. The right tibiae were harvested, fixed and embedded in methylmethacrylate for structural histomorphometric bone analysis at the proximal tibial metaphysis. The specific COX-2 or sPLA2 inhibitors prevented ovariectomy-induced (OVX-induced) decreases in trabecular connectivity (P<0.05); suppressed the acceleration of bone resorption; and maintained bone turnover at SHAM levels following OVX in the rat. The sPLA2 inhibitor significantly suppressed increases in osteoclast surface induced by OVX (P<0.05), while the effect of COX-2 inhibition was less marked. These findings demonstrate that inhibitors of COX-2 and sPLA2-IIa can effectively suppress OVX-induced bone loss in the adult rat by conserving trabecular bone mass and architecture through reduced bone remodeling and decreased resorptive activity. Moreover, we report an important role of sPLA2-IIa in osteoclastogenesis that may be independent of the COX-2 metabolic pathway in the OVX rat in vivo. PMID- 16473055 TI - Workshop on Intestinal Failure, February 7-8, 2004, Las Vegas, Nevada, USA. PMID- 16473056 TI - Short-chain fatty acids impact on intestinal adaptation, inflammation, carcinoma, and failure. PMID- 16473057 TI - Glutamine: do the data support the cause for glutamine supplementation in humans? AB - This review examines the preclinical rationale for using glutamine supplements and reviews the prospective randomized trials using glutamine to improve outcomes in patients. A special role for glutamine in gut physiology and in management of a variety of serious illnesses has been suggested, because it is the most abundant extracellular amino acid, and is used at high rates by the gut, liver, central nervous system, and immune cells. A state of relative Gln deficiency has been postulated in humans based on the decrease in plasma Gln in acute critical illness, but the decrease in plasma Gln is not specific for that amino acid, predicts only poorer outcome, and has not been validated to identify a deficiency state. Current evidence does not necessarily predict a special need or role for Gln in critical illness. Clinical efficacy of supplemental Gln has been difficult to demonstrate, possibly related to the lack of a Gln deficiency state, the wide range of end points used that reflect the lack of certainty of the predicted effect of supplementation, the heterogeneous patient populations studied, the lack of stable clinical course during the study, the lack of adequate power, and the relatively short follow-up period. Prospective randomized clinical trials of Gln supplementation were reviewed in patients with short-bowel syndrome, during cancer chemotherapy and in bone marrow transplantation, and in surgical, burn, and intensive care unit patients. No firm recommendation can be made at this time. Future studies should seek to develop a more standard and stable design for intervention in sufficiently powered studies. PMID- 16473058 TI - Novel agents in the treatment of intestinal failure: humoral factors. PMID- 16473060 TI - Glucagon-like peptide-2: update of the recent clinical trials. AB - Although long-term parenteral nutrition is lifesaving in patients with intestinal failure, it is expensive and associated with serious complications such as catheter sepsis, venous occlusions, and liver failure and severely impairs the quality of life in the short bowel patients. Therefore, treatments that increase the absolute intestinal absorption, thereby eliminating or minimizing the need for parenteral support, are needed. In this respect, glucagon-like peptide 2 (GLP 2) has received attention. In this review, the nature of the short bowel syndrome is described, and the antisecretory, transit-modulating, but also intestinotrophic effects of GLP-2 are presented. As illustrated in 2 pilot studies, one using native GLP-2 and the other a degradation-resistant analogue, teduglutide, these new agents may prove important in optimizing remnant intestinal function, thereby eliminating the need for parenteral support and improving quality of life in short bowel patients with intestinal failure. PMID- 16473059 TI - Short bowel syndrome: recent clinical outcomes with growth hormone. AB - There has been recent clinical enthusiasm for treating short bowel patients with either human recombinant growth hormone alone or in combination with glutamine and diet in hopes of reducing their parenteral nutrition requirements. Part of this interest has been the result of studies done in short bowel animal models showing increased bowel growth and function following such treatment. Since 1995, 5 key clinical studies have been published in peer-reviewed literature reporting the effects of growth hormone alone or in combination with glutamine and diet in patients with short bowel syndrome. Three of these controlled studies have reported negative results, and 1 controlled and 1 uncontrolled study reported positive results. This review discusses each of these articles and provides some explanation on how these studies and results may have differed. PMID- 16473062 TI - From the cradle to enteral autonomy: the role of autologous gastrointestinal reconstruction. AB - The short bowel state is treatable, with acceptable long-term quality of life. Management during the first 6-12 months of life is critical but, presently, frequently compromises long-term survival and prospects. At first presentation, primary caregivers, working with specialists at designated intestinal failure centers, should develop a structured individual-oriented management plan. Preservation of venous access, "hepatosparing" parenteral nutrition, and avoidance of liver sepsis are crucial to survival. Early surgery should be limited to conservation of autologous bowel, even short bowel lengths having great potential, and to facilitating natural intestinal adaptation. Bowel expansion may be relevant prior to delayed bowel reconstruction with single or combined techniques that include bowel lengthening and/or tailoring, reversed segments, and colon interposition. Bowel transplantation, as yet not recommended for primary management, offers survival and opportunity to those with no prospect of autologous bowel autonomy or following failed autologous gastrointestinal reconstruction. This paper reviews current surgery for the short bowel state and concludes that it is presently appropriate before bowel transplantation to offer autologous gastrointestinal reconstruction, with its prospect of enteral autonomy with quality life. It emphasizes the need for an individual-oriented management plan, developed jointly at the time of first presentation between the primary caregivers and a designated multidisciplinary intestinal failure center, to enhance the prospects for enteral autonomy preferably on autologous bowel. Autologous gastrointestinal reconstruction is in its infancy and requires resources, commitment, and research from dedicated bowel reconstructive surgeons toward a better opportunity for the child and family with short bowel. PMID- 16473061 TI - Intestinal transplantation for short bowel syndrome and gastrointestinal failure: current consensus, rewarding outcomes, and practical guidelines. PMID- 16473063 TI - Intestinal replacement therapy: timing and indications for referral of patients to an intestinal rehabilitation and transplant program. AB - Current treatment options for patients suffering from intestinal insufficiency include all forms of intestinal replacement therapy (IRT). Parenteral nutrition has achieved extended success for the majority of patients requiring interval treatment, however, complications leading to failure of this treatment increases with the duration of therapy. There is currently no consensus as to the appropriate timing for transplantation of the intestine or the timing of referral for evaluation at a center experienced with this therapy. Certain patient characteristics warrant evaluation. Those patients with no jejunoileum who have guaranteed lifelong parenteral dependence, both adult and pediatric, should be immediately referred to a transplant center due to the high likelihood of the development of liver disease. Patients with metastatic infectious complications from catheter sepsis, patients with cholestasis seen intermittently with sepsis episodes, patients who are not successfully weaning and who demonstrate progressive thrombocytopenia, and patients with motility disorder experiencing deterioration should also warrant early referral to an intestinal rehabilitation and transplant program. The objective of evaluation is to maximize the opportunities for rehabilitation while not missing the critical window of opportunity for successful transplantation when needed. We favor an aggressive directed approach to rehabilitation, coupled with psychological preparation for both transplantations and other options. Early referral requires trust between the patient, referring physician, and the transplant team to assure that a rush to judgment will not lead to a premature transplant. The current wait list mortality is high, mandating early referral and listing with an approach aimed at maximizing both the success of gastrointestinal support, as well as of transplantation when necessary. PMID- 16473064 TI - Innovative approaches to improving organ availability for small bowel transplant candidates. PMID- 16473065 TI - Cost and quality of life after intestinal transplantation. AB - Intestinal transplantation has become a standard treatment for intestinal failure in patients with life-threatening complications of TPN. Although the long-term survival of patients with continued parenteral nutrition is higher than after intestinal transplantation, the 1 and 2 year survival is comparable. Here we examine other aspects of the treatment options available for patients with intestinal failure including the cost of the therapy and the quality of life. The cost of parenteral nutrition compared to intestinal transplantation reveals that transplantation is cost-effective in patients that maintain graft function within 1 to 3 years after surgery. The quality of life after transplantation is probably equal to or better than quality of life on TPN and children report quality of life similar to normal school children. Although currently reserved for those with life-threatening complications, intestinal transplantation may soon be an option for any patient permanently dependent on parenteral nutrition. PMID- 16473066 TI - Causes and management of intestinal failure in children. AB - Intestinal failure is a condition requiring the use of parenteral nutrition as long as it persists. Causes of severe protracted intestinal failure include short bowel syndrome, congenital diseases of enterocyte development, and severe motility disorders (total or subtotal aganglionosis or chronic intestinal pseudo obstruction syndrome). Intestinal failure may be irreversible in some patients, thus requiring permanent parenteral nutrition. Liver disease may develop with subsequent end-stage liver cirrhosis in patients with intestinal failure as a consequence of both underlying digestive disease and unadapted parenteral nutrition. Death will occur if combined liver-intestine transplantation is not performed. Catheter-related sepsis and/or extensive vascular thrombosis may impede the continuation of a safe and efficient parenteral nutrition and may also require intestinal transplantation in some selected cases. Thus management of patients with intestinal failure requires an early recognition of the condition and the analysis of its risk of irreversibility. Timing of referral for intestinal transplantation remains a crucial issue. As a consequence, management should include therapies adapted to each stage of intestinal failure based on a multidisciplinary approach in centers involving pediatric gastroenterology, parenteral nutrition expertise, home parenteral nutrition program, pediatric surgery, and liver intestinal transplantation program. PMID- 16473067 TI - Optimal management of the post-intestinal transplant patient. AB - The lack of controlled interventional studies limits the ability to assess optimal management of intestine transplant recipients. This report aims to examine factors that probably impact on the quality of patient care in the setting of intestine transplantation. The specific practice in the most experienced intestine transplant programs in the United States was surveyed with regard to immunosuppressive regimens, treatment of acute allograft rejection, feeding, and viral surveillance and treatment. The most striking finding was in the level of agreement between the centers, particularly with regard to use of tacrolimus for maintenance immunosuppression, methylprednisone boluses for treatment of acute rejection, early postoperative enteral feeding, and ganciclovir prophylaxis. PMID- 16473068 TI - Chronic intestinal pseudo-obstruction: assessment and management. PMID- 16473069 TI - Overview of etiology and management of intestinal failure. PMID- 16473070 TI - Parenteral nutrition: transient or permanent therapy in intestinal failure? PMID- 16473071 TI - Guidelines for management of home parenteral support in adult chronic intestinal failure patients. AB - Management of home parenteral support in adult benign but chronic intestinal failure patients requires a nutrition support team using disease-specific pathways. Education of patients to ensure they self manage home parenteral nutrition (HPN) is cornerstone to obtain minimal rate of technical complications and improvement in quality of life. Nutritive mixtures, compounded by pharmacists in single "all-in-one" bags, must be tailored according to the nutritional and intestinal status of individual patients with definition of macronutrients and water-electrolyte needs, respectively. Each PN cycle should be complete in essential nutrients to be nutritionally efficient and should have sufficient amounts of amino acids, dextrose, water, minerals, and micronutrients to avoid deficiency. When the nutritional goal is achieved, a minimum number of PN cycles per week should be implemented, guided ideally by digestive balance(s) (In-Out) of macronutrients and minerals of individual patients. Indeed, HPN is, in most cases, a complementary nonexclusive mode of nutritional support. In short gut patients--who represent 75% of chronic intestinal failure patients--encouraging enteral feeding decrease PN delivery and the risk of metabolic liver disease associated with HPN. In short gut patients with no severe renal impairment, blood citrulline dosage, in association with the remnant anatomy, is a tool to delineate transient from permanent intestinal failure. The latter group includes candidates for trophic gut factors and rehabilitative or reconstructive surgery, including intestinal transplantation. Thus, outcome improvement for intestinal failure patients needs intestinal failure teams having expertise in all medical and surgical aspects of this field. PMID- 16473072 TI - Etiology and initial management of short bowel syndrome. PMID- 16473073 TI - Home parenteral nutrition: survival, cost, and quality of life. AB - This article summarizes what is known about which factors influence survival of patients on home parenteral nutrition, the costs related to this therapy, and the quality of life for patients living on home parenteral nutrition. The article refers to both North American and European experiences with this complex therapy. PMID- 16473074 TI - Management of short bowel syndrome: avoidance of total parenteral nutrition. PMID- 16473076 TI - Intestinal failure-associated liver disease: what do we know today? AB - Intestinal failure-associated liver disease develops in 40% to 60% of infants who require long-term total parenteral nutrition (TPN) for intestinal failure and 15% to 40% of adults on home parenteral nutrition. The clinical spectrum includes hepatic steatosis, cholestasis, cholelithiasis, and hepatic fibrosis. Progression to biliary cirrhosis and the development of portal hypertension and liver failure occurs in a minority but is more common in infants and neonates than in adults. The pathogenesis is multifactorial. In infants it is related to prematurity, low birth weight, duration of PN, short bowel syndrome requiring multiple laparotomies, and recurrent sepsis. Other important mechanisms include lack of enteral feeding, which leads to reduced gut hormone secretion; reduction of bile flow and biliary stasis, which leads to the development of cholestasis; and biliary sludge and gallstones, which exacerbate hepatic dysfunction. In adults, IFALD is less common and related to age, length of time on PN, total caloric intake, and lipid or glucose overload. In preterm infants, a deficiency of taurine or cysteine may play a role, whereas in both adults and children, choline deficiency may exacerbate IFALD. Lipid emulsions, choline deficiency, and manganese toxicity are associated with both hepatic steatosis and cholestasis in adults and children. Management strategies for the prevention of intestinal failure-induced liver disease include early enteral feeding, a multidisciplinary approach to the management of parenteral nutrition, and aseptic catheter techniques to reduce sepsis. The addition of choline, taurine, and cysteine to PN solutions may also play a role. Oral administration of ursodeoxycholic acid may improve bile flow and reduce gallbladder stasis. Survival after either isolated small bowel or combined liver and small bowel transplantation is approximately 50% at 5 years, making this an acceptable therapeutic option in adults and children with irreversible liver and intestinal failure. PMID- 16473075 TI - Bacterial overgrowth and liver complications in short bowel intestinal failure patients. PMID- 16473077 TI - Small intestinal bacterial overgrowth: roles of antibiotics, prebiotics, and probiotics. AB - Small intestinal bacterial overgrowth is common in intestinal failure. Its occurrence relates to alterations in intestinal anatomy, motility, and gastric acid secretion. Its presence may contribute to symptoms, mucosal injury, and malnutrition. Relationships between bacterial overgrowth and systemic sepsis are of potential importance in the intestinal failure patient because the direct translocation of bacteria across the intestinal epithelium may contribute to systemic sepsis: a phenomenon that has been well established in experimental animal models. The accurate diagnosis of bacterial overgrowth continues to present a number of challenges in clinical practice and especially so among patients with intestinal failure. The management of patients with bacterial overgrowth remains, for the most part, primarily empiric and comprises antibiotic therapy and correction of any associated nutritional deficiencies. Although evidence from experimental animal studies consistently indicates that probiotics exert barrier-enhancing, antibacterial, immune-modulating, and anti-inflammatory effects, which all could be benefits in small intestinal bacterial overgrowth and intestinal failure, their role in human beings remains to be evaluated adequately. PMID- 16473078 TI - Specific nutrients in intestinal failure: one size fits no one. PMID- 16473079 TI - Mechanisms of enteral nutrient-enhanced intestinal adaptation. AB - The role of enteral nutrients in maintaining small intestinal structure and function is well established. Evidence that enteral nutrients induce intestinal adaptation include the structural and functional gradient along the length of the healthy intestine, the atrophy and functional compromise induced by fasting and parenteral nutrition, and the enhanced adaptive capacity of the distal intestine following partial enterectomy. Key mechanisms contributing to enteral nutrient induced intestinal adaptation include nonspecific luminal stimulation and that provided by specific nutrients, "functional workload" induced by polymeric nutrients, potential stimulation of pancreaticobiliary secretions, secretion of humoral mediators, and induction of intestinal hyperemia. PMID- 16473080 TI - The neutral zone in complete dentures. 1976. PMID- 16473082 TI - Surgical and prosthetic planning for a two-implant-retained mandibular overdenture: a clinical report. AB - A 2-implant-retained mandibular overdenture is considered by some to be the standard of care for mandibular edentulism. Compared to a conventional complete denture, an implant-retained overdenture requires more thorough planning. Careful consideration is necessary regarding the 3-dimensional orientation of the implants to ensure adequate horizontal and vertical space for prosthetic components. This clinical report describes a patient with a compromised mandibular overdenture in whom the position of the existing implants yielded insufficient space for prosthetic components. This report describes the concepts for treatment planning prior to fabricating a new mandibular overdenture, including considerations for the surgical removal of the existing implants, alveoloplasty to create the necessary space for prosthetic components, and placement of the new implants to ensure an esthetic and functional prosthesis. PMID- 16473083 TI - Osteomyelitis and pathological mandibular fracture related to a late implant failure: a clinical report. AB - This clinical report describes an unusual consequence of peri-implantitis and subsequent implant failure in a 72-year-old man. Following uneventful implant removal, osteomyelitis developed at the implant site, which subsequently weakened the mandible sufficiently for a pathological fracture to occur. The patient was treated successfully by use of an intensive oral hygiene regime, antibiotics, and conservative management. The consequences and management of implant failure are discussed, as well as issues relating to the management of osteomyelitis and pathological bone fracture. PMID- 16473084 TI - Fracture resistance of single-tooth implant-supported all-ceramic restorations: an in vitro study. AB - STATEMENT OF PROBLEM: High-strength ceramic materials can be used to fabricate esthetic and stable implant-supported single-tooth restorations. No study was identified that compared the fracture resistance of individual components of single-tooth implant-supported all-ceramic restorative systems after artificial aging. PURPOSE: The purpose of this in vitro study was to evaluate the fracture resistance of single-tooth implant-supported all-ceramic restorations consisting of alumina all-ceramic restorations on different implant abutments and to identify the weakest component of the restorative system. MATERIAL AND METHODS: Forty-eight standardized maxillary central incisor alumina crowns (Procera) were fabricated for each of the 3 test groups (n = 16) (Control group Ti, titanium abutments; Group Al, alumina abutments; Group Zr, zirconia abutments) for the Replace implant system. The crowns were adhesively luted using a resin luting agent (Panavia 21) and artificially aged through dynamic loading and thermal cycling. Afterwards, all specimens were tested for fracture resistance using compressive load on the palatal surfaces of the crowns. Kruskal-Wallis analysis of variance and post hoc Wilcoxon rank sum tests were performed to test for differences in fracture resistance values (alpha = .05). RESULTS: All test specimens survived the artificial aging process using simulated oral conditions. No screw loosening was recorded. The median fracture resistance was 1454 N, 422.5 N, and 443.6 N for groups Ti, Al, and Zr, respectively. Significant differences were found for the fracture resistance comparisons of group Ti with groups Al and Zr (Kruskal-Wallis test, P < .001). The test results for the comparison of groups Al and Zr were not significant. CONCLUSION: All 3 implant-supported restorations have the potential to withstand physiologic occlusal forces applied in the anterior region. PMID- 16473085 TI - Fracture load of composite resin and feldspathic all-ceramic CAD/CAM crowns. AB - STATEMENT OF PROBLEM: Various machinable materials are currently used with computer-aided design/computer-assisted manufacturing (CAD/CAM) technologies for the chairside fabrication of restorations. However, properties of these new machinable materials, such as fracture load, wear, marginal deterioration, and color stability, should be investigated in vitro under replicated clinical conditions prior to time-consuming clinical studies. PURPOSE: This study investigated the effect of cyclic loading fatigue and different luting agents under wet conditions on the fracture load of CAD/CAM machined composite resin and all-ceramic crowns. MATERIALS AND METHODS: Ninety-six intact human maxillary premolars were prepared for composite resin and all-ceramic crowns with the following preparation criteria: 6-degree axial taper, 1.5-mm shoulder finish line placed 0.5 mm occlusal to the cemento-enamel junction, 1.5-mm axial reduction, 2 mm occlusal reduction, and 5-mm occluso-gingival height. Sixteen unprepared premolars served as controls. Forty-eight all-ceramic crowns (Vita Mark II) and 48 millable composite resin crowns (MZ100 Block) were fabricated using a CAD/CAM system (Cerec 3). Three luting agents-RelyX ARC (RX), GC Fuji CEM (FC), and zinc phosphate cement (ZP)-were used for cementation (n = 16). After 1-week storage in water, half of the specimens (n=8) in each subgroup were cyclically loaded and thermal cycled under wet conditions for 600,000 masticatory cycles and 3500 thermal cycles (58 degrees C/4 degrees C; dwell time, 60 seconds) in a masticatory simulator; the other half (n = 8) were fractured without cyclic loading. All specimens were loaded in a universal testing machine with a compressive load (N) applied along the long axis of the specimen at a crosshead speed of 1 mm/min until fracture. Fracture loads (N) were recorded for each specimen. Three-way analysis of variance was used to detect the effects of the experimental factors (crown material, luting agent, and loading conditions) on the fracture load. The comparison with the unprepared natural teeth as controls was done by means of t tests (alpha=.05). RESULTS: Analysis of variance revealed a statistically significant influence of the luting agent and the cyclic loading (P < .001), whereas the crown material had no significant influence. Cyclic loading fatigue significantly decreased the mean fracture load of test groups independent of the 3 luting agents used: MZ100/ZP, 827.1 to 552.5 N; MZ100/FC, 914.7 to 706.2 N; MZ100/RX, 955.9 to 724.4 N; Vita/ZP, 772.3 to 571.5 N; Vita/FC, 923.6 to 721.1 N; and Vita/RX, 929.1 to 752.7 N. However, there was no significant difference in the mean fracture load of control specimens before and after cyclic loading (1140.1 N and 1066.2 N, respectively). Adhesive luting agents RelyX ARC and GC Fuji CEM increased fracture load significantly compared to zinc phosphate cement. CONCLUSIONS: Cyclic loading fatigue significantly reduced the fracture loads of composite resin and all-ceramic crowns, whereas adhesive cementation significantly increased the fracture loads. PMID- 16473086 TI - Effect of core and veneer thicknesses on the color parameters of two all-ceramic systems. AB - STATEMENT OF PROBLEM: Specific contributions of the core and the veneer thickness on the appearance of layered disk specimens are not well characterized. PURPOSE: This study examined the effect of varying core and veneer thickness on the color parameters of layered disk specimens made of 2 ceramic systems. MATERIAL AND METHODS: Disk specimens 16 mm in diameter with core/veneer thickness (mm) of 0.8/0.2 (baseline), 0.8/0.7, 0.8/1.2, 1.0/0.5, 1.3/0.2, and 1.8/0.2 were made from a leucite-reinforced ceramic (IPS Empress; 2B shade), and thickness (mm) of 0.5/0.5 (baseline), 0.5/1.0, 0.5/1.5, 0.8/0.7, 1.0/0.5, and 1.5/0.5 were made from a glass-infiltrated spinell ceramic (In-Ceram Spinell; A2 shade). Color parameters L*, a*, and b* of CIELAB color space were measured against a neutral gray background with a tri-stimulus colorimeter, and deltaE between disk group and its respective baseline group was calculated. Analysis of variance was used to determine the effect of material and core and veneer thickness on the color parameters (alpha = .05). RESULTS: Increasing the total disk thickness resulted in decrease of L* (P < .001) and an increase of a* (P < .001), b* (P < .001), and deltaE (P = .0236). For leucite-reinforced ceramic, the core thickness, veneer thickness, and their interaction exhibited significant influence on the mean values of a* and b* (P < .0028) but not on the mean values of L*. For spinell ceramic, the core thickness and veneer thickness exhibited significant influence on the mean values of L* and b* (P < .0272) but not on a*; the interaction demonstrated significant influence on b* (P = .0003) but not on L* and a*. CONCLUSION: The color appearance of the layered ceramic disk specimens is strongly influenced not only by the core thickness and veneer thickness, but also by their interaction. PMID- 16473087 TI - Resistance of three new esthetic post-and-core systems to compressive loading. AB - STATEMENT OF PROBLEM: Despite their esthetic properties, the fracture resistance of newly introduced esthetic post-and-core systems has not been studied, restricting clinical use. PURPOSE: The purpose of this study was to determine and compare the fracture resistances of 3 recently introduced esthetic post-and-core systems with a cast metal post and core using a clinically related test method. MATERIAL AND METHODS: Forty recently extracted caries-free maxillary central incisors were selected, sectioned, and their roots endodontically treated. The roots were then embedded in acrylic resin blocks and assigned to 4 experimental groups (n = 10). The cast metal (Wiron 99) post-and-core group (CMDC) served as the control. Three experimental groups consisted of a stainless steel post/composite-resin (Charisma) core group (SCOM), zirconium dioxide post/composite-resin (Charisma) core group (ZCOM), and zirconium dioxide post/ceramic (Cosmopost) core group (ZCER). The post spaces were prepared, posts were seated, cores were formed, and 40 post-and-core foundations were cemented into the roots. Following thermal cycling (5000 cycles between 5 degrees C and 55 degrees C with a dwell time of 30 seconds), a compressive load was applied to the inclined surface of each specimen at a 130-degree angle to the specimen long axes in a universal testing machine at a crosshead speed of 1 mm/min until failure occurred. Force to failure (N) was recorded. Data were analyzed with a 1-way analysis of variance and Bonferroni tests to determine the difference between the groups (alpha=.05). For pairwise comparisons among group mean values, P < .0083 indicated statistical significance and established the overall confidence level at 95%. RESULTS: Groups CMDC (738.6 N +/- 10.2) and ZCER (710.2 N +/- 5.3) exhibited the highest resistance to fracture. The ZCOM group showed the lowest fracture resistance (450.3 N +/- 5.7). The fracture resistance of the SCOM group (565 N +/- 7.2) was lower than the CMDC and ZCER groups, and higher than the ZCOM group. The differences among the groups were significant (P < .001) except for CMDC and ZCER (P < .016). CONCLUSION: Within the limitations of this study, the cast metal post/core and zirconia post/ceramic core foundations were found to be more fracture resistant than the zirconia post/composite-resin core and stainless steel post/composite-resin core foundations. Aside from its desirable esthetic properties, the zirconia post/ceramic core combination demonstrated high resistance to fracture. PMID- 16473088 TI - Effects of staining and bleaching on color change of dental composite resins. AB - STATEMENT OF PROBLEM: Discoloration of resin-based composites by colored solutions is a common problem. The use of bleaching agents for discolored natural teeth is becoming increasingly popular. It is not clear if bleaching agents can remove the stain from composite resins. PURPOSE: The purpose of this study was to investigate the effects of 2 staining solutions and 3 bleaching systems on the color changes of 2 dental composite resins. MATERIAL AND METHODS: Forty-five disk shaped specimens (9 x 2.5 mm) of each of 2 composite resins, Filtek Supreme (FS) and Esthet X (EX), were prepared. The specimens were then divided into 3 groups of 15 specimens each and immersed in 2 staining solutions (coffee or red wine) or distilled water (control) for 3 hours daily over a 40-day test period. The 3 groups were then divided into 3 subgroups (n = 5), and 3 bleaching agents (Crest Night Effects, Colgate Simply White Night, or Opalescence Quick) were applied to the surface of the specimens over a 14-day period. Color of the specimens was measured with a spectrophotometer using CIELAB color space relative to CIE standard illuminant D55 at baseline, after staining, and after bleaching. The color differences (deltaE(ab)*) between the 3 measurements were calculated. The value deltaE(ab)* = 3.3 was used as an acceptable value in subjective visual evaluations. Analysis of variance and nonparametric analysis (Kruskal-Wallis test and Mann-Whitney test) were used to analyze the data. RESULTS: After staining, FS had more color change than EX and was more affected by the wine solution. After bleaching, the color of both EX and FS specimens returned to the baseline. The color differences between bleaching and baseline were less than value deltaE(ab)* = 3.3 for all groups. CONCLUSION: The nanocomposite (FS) changed color more than the microhybrid composite (EX) as a result of staining in coffee or red wine solutions. After bleaching, discoloration was removed completely from the composite resins tested. PMID- 16473089 TI - The evaluation of microleakage and bond strength of a silicone-based resilient liner following denture base surface pretreatment. AB - STATEMENT OF PROBLEM: The failure of adhesion between a silicone-based resilient liner and a denture base is a significant clinical problem. PURPOSE: The purpose of this study was to examine the effects of denture base resin surface pretreatments with different chemical etchants preceding the silicone-based resilient liner application on microleakage and bond strength. The initial effects of chemical etchants on the denture base resin in terms of microstructural changes and flexural strength were also examined. MATERIAL AND METHODS: Forty-two polymethyl methacrylate (PMMA) denture base resin (Meliodent) specimens consisting of 2 plates measuring 30 x 30 x 2 mm were prepared and divided into 7 groups (n = 6). Specimen groups were treated by immersion in acetone for 30 (A30) or 45 (A45) seconds, methyl methacrylate monomer for 180 (M180) seconds, and methylene chloride for 5 (MC5), 15 (MC15) or 30 (MC30) seconds. Group C had no surface treatment and served as the control. Subsequently, an adhesive (Mollosil) and a silicone-based resilient denture liner (Mollosil) were applied to the treated surfaces, and all specimens were immersed in the radiotracer solution (thalium-201 chloride) for 24 hours. Tracer activity (x-ray counts), as a parameter of microleakage, was measured using a gamma camera. For bond-strength measurement, 84 rectangular PMMA specimens (10 x 10 x 40 mm) were surface-smoothed for bonding and treated with the different chemical etchants using the same previously described group configurations. The adhesive and the silicone-based denture liner were applied to the treated surfaces. Tensile bond-strength (MPa) was measured in a universal testing machine. Flexural strength measurement was performed with 49 PMMA specimens (65 x 10 x 3.3 mm according to ISO standard 1567) in 7 groups (n = 7), with 1 flat surface of each treated with 1 of the chemical etchants preceding adhesive application. The flexural strength (MPa) was measured using a 3-point bending test in a universal testing machine. The data were analyzed by 1-way analysis of variance and the Tukey HSD test (alpha = .05). RESULTS: Significant differences were found between the groups in terms of microleakage (P < .0001). The lowest microleakage was observed in group M180 (30,000 x-ray counts) and the highest in the control group (44,000 x-ray counts). The mean bond strength to PMMA resin ranged from 1.44 to 2.22 MPa. All treated specimens showed significantly higher bond strength than controls (P < .01). The flexural strength values all significantly differed (P < .05). All experimental specimens that had chemical surface treatments showed lower flexural strength than controls (P < .05). CONCLUSION: Treating the denture base resin surface with chemical etchants increased the bond strength of silicone based resilient denture liner to denture base and decreased the microleakage between the 2 materials. Considering the results of both tests together, the use of methyl methacrylate monomer for 180 seconds was found to be the most effective chemical treatment. PMID- 16473090 TI - Evidence-based decision making: Guide to reading the dental materials literature. AB - Although potential links between materials data and clinical behavior are often implied, the status of such linkage is often left obscure. This paper provides clinicians a context within which to view materials information as evidence for clinical indications and to broaden readers' appreciation for the subject. Hierarchies of both clinical and nonclinical data are presented and discussed from the point of view of their predictive potential regarding clinical performance. Excellent sources of information are identified for the clinician making treatment decisions, and perspectives are offered on the value of other published materials data. PMID- 16473091 TI - Immediate function in the edentulous mandible: replacement of a lost implant using prefabricated guides. AB - This article describes a technique for replacement of a lost implant. The procedure involves the use of templates, drill guides, and drills of the system to replace a 4.5-mm-wide lost implant with a 5.0-mm-wide implant. The surgical procedure was simplified to optimize the healing process and to be more comfortable for the patient. The accuracy of the templates and guides allowed for ideal position of the implant and the immediate use of the original fixed implant supported prosthesis. PMID- 16473092 TI - A technique for locating implant abutment screws of posterior cement-retained metal-ceramic restorations with ceramic occlusal surfaces. AB - With the increased use of cement-retained implant-supported restorations for the replacement of missing teeth, clinicians may choose to use a definitive cement to lute the definitive restoration. A complication that may occur, especially for a single-tooth replacement, is loosening of the abutment screw. In those situations, it may be difficult to locate the abutment-screw access to remove the restoration. The purpose of this article is to describe a technique that may facilitate the clinician's ability to locate the abutment-screw access in the event of abutment-screw loosening, thus reducing the need for refabricating the restoration. PMID- 16473093 TI - An investment technique for custom dental implant abutments. PMID- 16473094 TI - A technique for salvaging an implant-supported crown with a fractured abutment screw. PMID- 16473095 TI - A photo guide for arranging artificial anterior teeth. PMID- 16473096 TI - Emergencies in the outpatient setting: Part II. PMID- 16473097 TI - Angina pectoris: evaluation in the office. AB - Angina pectoris is a clinical manifestation of myocardial ischemia. Complete evaluation consists of a review of risk factors, a careful history, and, typically, a provocative test. Stress testing can be performed with exercise(treadmill, bicycle, or arm ergometry) or pharmacologic agents that increase cardiac work (dobutamine) or dilate the coronary vessels (adenosine or dipyridamole). Patients who have high-risk features found by clinical history or by stress testing should be referred for coronary angiography and possible revascularization. Comprehensive management of patients who have angina (with or without revascularization) includes smoking cessation,diet and weight control, vasculoprotective drugs (aspirin, statins, and possibly ACE inhibitors), and antianginal medications (nitrates, D-blockers, and calcium channel blockers). These strategies have led to an important reduction in morbidity and mortality over the past 2 decades, and the focus on implementing guidelines for patients who are currently undertreated is expected to improve outcomes further. PMID- 16473098 TI - Arrhythmias in the office. AB - The incidence of patients who present to the office with arrhythmia and hemodynamic instability is unknown. Emergency medical systems data, based on ambulance runs, are available only for patients who have had a cardiac arrest. When faced with an unstable or potentially unstable patient, however, we must be prepared to act quickly, safely, and accurately. This article addresses the general approach to such a patient; provides necessary information on office emergency preparation, including training, rapid response team protocol, and the use of automated external defibrillators; and addresses the identification and initial office management of the various rhythms that are capable of threatening a patient's life. PMID- 16473099 TI - Hypertensive emergency and severe hypertension: what to treat, who to treat, and how to treat. AB - Remember to treat patients, not numbers. Use fast acting shortterm medicines only when convincing evidence of rapidly evolving end-organ damage is present. For all patients, emergent or asymptomatic, the treatment goal is long-term control of hypertension. Potent IV agents for the im-mediate control of elevated blood pressure need to be used cautiously,bearing in mind both the side effects and the hazards of overly rapid control of hypertension. Conventional oral medication regimens demonstrated to modify the risks of chronic hypertension should be used whenever possible and as early as is practical to promote gradual control of hypertension. Whenever a patient presents for the evaluation of severe hypertension in an emergent setting, take the opportunity to encourage appropriate ongoing follow-up; after all, hypertension is not a single episode, it is an ongoing threat to good health. PMID- 16473100 TI - Dyspnea. AB - When evaluating a dyspneic patient in the office, a quick initial assessment of the airway, breathing, and circulation, while gathering a brief history and focused physical examination are necessary. Most often, an acute cardiopulmonary disorder, such as CHF, cardiac ischemia, pneumonia, asthma, or COPD exacerbation, can be identified and treated. Stable patients who improve can be sent home, but those in acute distress with unstable or impending unstable conditions need to be transferred emergently to definitive care. Because of the difficult logistics involved in attempting to work up an outpatient for new onset of SOB, some patients will need to be transferred to the nearest ED for a definitive diagnosis. PMID- 16473101 TI - Acute abdominal pain. AB - Because there are many causes of acute abdominal pain, a systematic approach by the evaluating physician is necessary to narrow the differential diagnosis. It is vital that the physician have an understanding of the mechanisms of pain generation and be familiar with the presentations of common diseases that cause abdominal pain. Recognizing the red flags in the history and physical examination and the initial imaging and laboratory findings helps to determine which patients may have a serious underlying disease process, and therefore warrant more expedited evaluation and treatment. PMID- 16473102 TI - Back pain emergencies. AB - Most adults in the United States will experience an episode of back pain at some point during their lifetime. Most will present to their primary care physician for evaluation and treatment. Many patients have non-life-threatening etiologies and recover within 4 to 6 weeks. A small percentage, however, have back pain due to a potentially life-threatening emergency. AD,rupturing AAA, SEM, cauda equina syndrome, vertebral osteomyelitis,and SEA are just some of the medical emergencies that can present with back pain. Clinical suspicion for these diagnoses begins with a thorough history and physical examination. It is imperative that the office-based physician search for and accurately identify any red flag within the history or physical examination. Appropriate laboratory studies and diagnostic imaging are obtained based on the suspected etiology. PMID- 16473103 TI - Corruption in health care costs lives. PMID- 16473104 TI - The value of a good nap. PMID- 16473105 TI - Research versus treatment for neglected diseases. PMID- 16473106 TI - What are the implications of optimism bias in clinical research? PMID- 16473107 TI - Rheumatoid arthritis: the goal rather than the health-care provider is key. PMID- 16473108 TI - Somatisation: a joint responsibility of doctor and patient. PMID- 16473109 TI - Nuclear receptors, bile-acid detoxification, and cholestasis. PMID- 16473110 TI - Postconditioning for protection of the infarcting heart. PMID- 16473111 TI - Research injury in clinical trials in South Africa. PMID- 16473112 TI - I'll scratch your back ... if you have one. PMID- 16473113 TI - Ernest McCulloch: father of experimental haematology. PMID- 16473114 TI - Face transplants. PMID- 16473115 TI - Efficacy and safety of cholesterol-lowering treatment. PMID- 16473116 TI - Efficacy and safety of cholesterol-lowering treatment. PMID- 16473119 TI - Guillain-Barre syndrome. PMID- 16473120 TI - Guillain-Barre syndrome. PMID- 16473121 TI - Guillain-Barre syndrome. PMID- 16473123 TI - Palliative care for management of small-cell lung cancer. PMID- 16473124 TI - Development of adenoviral-vector-based pandemic influenza vaccine against antigenically distinct human H5N1 strains in mice. AB - INTRODUCTION: Avian H5N1 influenza viruses currently circulating in southeast Asia could potentially cause the next pandemic. However, currently licensed human vaccines are subtype-specific and do not protect against these H5N1 viruses. We aimed to develop an influenza vaccine and assessed its immunogenicity and efficacy to confer protection in BALB/c mice. METHODS: We developed an egg independent strategy to combat the avian influenza virus, because the virus is highly lethal to chickens and the maintenance of a constant supply of embryonated eggs would be difficult in a pandemic. We used a replication-incompetent, human adenoviral-vector-based, haemagglutinin subtype 5 influenza vaccine (HAd-H5HA), which induces both humoral and cell-mediated immune responses against avian H5N1 influenza viruses isolated from people. FINDINGS: Immunisation of mice with HAd H5HA provided effective protection from H5N1 disease, death, and primary viral replication (p<0.0001) against antigenically distinct strains of H5N1 influenza viruses. Unlike the recombinant H5HA vaccine, which is based on a traditional subunit vaccine approach, HAd-H5HA vaccine induced a three-fold to eight-fold increase in HA-518-epitope-specific interferon-gamma-secreting CD8 T cells (p=0.01). INTERPRETATION: Our findings highlight the potential of an Ad-vector based delivery system, which is both egg-independent and adjuvant-independent and offers stockpiling options for the development of a pandemic influenza vaccine. PMID- 16473125 TI - Incidence of clinically significant bacteraemia in children who present to hospital in Kenya: community-based observational study. AB - BACKGROUND: Estimates of the burden of invasive bacterial disease in sub-Saharan Africa have previously relied on selected groups of patients, such as inpatients; they are, therefore, probably underestimated, potentially hampering vaccine implementation. Our aim was to assess the incidence of bacteraemia in all children presenting to a hospital in Kenya, irrespective of clinical presentation or decision to admit. METHODS: We did a community-based observational study for which we cultured blood from 1093 children who visited a Kenyan hospital outpatient department. We estimated bacteraemia incidence with a Demographic Surveillance System, and investigated the clinical significance of bacteraemia and the capacity of clinical signs to identify cases. RESULTS: The yearly incidence of bacteraemia per 100,000 children aged younger than 2 years and younger than 5 years was 2440 (95% CI 1307-3573) and 1192 (692-1693), respectively. Incidence of pneumococcal bacteraemia was 597 (416-778) per 100,000 person-years of observation in children younger than age 5 years. Three-quarters of episodes had a clinical focus or required admission, or both; one in six was fatal. After exclusion of children with occult bacteraemia, the incidence of clinically significant bacteraemia per 100,000 children younger than age 2 years or 5 years fell to 1741 (790-2692) and 909 (475-1343), respectively, and the yearly incidence of clinically significant pneumococcal bacteraemia was 436 (132 739) per 100,000 children younger than 5 years old. Clinical signs identified bacteraemia poorly. INTERPRETATION: Clinically significant bacteraemia in children in Kilifi is twice as common, and pneumococcal bacteraemia four times as common, as previously estimated. Our data support the introduction of pneumococcal vaccine in sub-Saharan Africa. PMID- 16473126 TI - Obstetric outcomes after conservative treatment for intraepithelial or early invasive cervical lesions: systematic review and meta-analysis. AB - BACKGROUND: Conservative methods to treat cervical intraepithelial neoplasia and microinvasive cervical cancer are commonly used in young women because of the advent of effective screening programmes. In a meta-analysis, we investigated the effect of these procedures on subsequent fertility and pregnancy outcomes. METHODS: We searched for studies in MEDLINE and EMBASE and classified them by the conservative method used and the outcome measure studied regarding both fertility and pregnancy. Pooled relative risks and 95% CIs were calculated with a random effects model and interstudy heterogeneity was assessed with Cochrane's Q test. FINDINGS: We identified 27 studies. Cold knife conisation was significantly associated with preterm delivery (<37 weeks; relative risk 2.59, 95% CI 1.80 3.72, 100/704 [14%] vs 1494/27 674 [5%]), low birthweight (<2500 g; 2.53, 1.19 5.36, 32/261 [12%] vs 905/13 229 [7%]), and caesarean section (3.17, 1.07-9.40, 31/350 [9%] vs 22/670 [3%]). Large loop excision of the transformation zone (LLETZ) was also significantly associated with preterm delivery (1.70, 1.24-2.35, 156/1402 [11%] vs 120/1739 [7%]), low birthweight (1.82, 1.09-3.06, 77/996 [8%] vs 49/1192 [4%]), and premature rupture of the membranes (2.69, 1.62-4.46, 48/905 [5%] vs 22/1038 [2%]). Similar but marginally non-significant adverse effects were recorded for laser conisation (preterm delivery 1.71, 0.93-3.14). We did not detect significantly increased risks for obstetric outcomes after laser ablation. Although severe outcomes such as admission to a neonatal intensive care unit or perinatal mortality showed adverse trends, these changes were not significant. INTERPRETATION: All the excisional procedures to treat cervical intraepithelial neoplasia present similar pregnancy-related morbidity without apparent neonatal morbidity. Caution in the treatment of young women with mild cervical abnormalities should be recommended. Clinicians now have the evidence base to counsel women appropriately. PMID- 16473127 TI - Epilepsy in children. AB - 10.5 million children worldwide are estimated to have active epilepsy. Over the past 15 years, syndrome-oriented clinical and EEG diagnosis, and better aetiological diagnosis, especially supported by neuroimaging, has helped to clarify the diversity of epilepsy in children, and has improved management. Perinatal and postinfective encephalopathy, cortical dysplasia, and hippocampal sclerosis account for the most severe symptomatic epilepsies. Ion channel defects can underlie both benign age-related disorders and severe epileptic encephalopathies with a progressive disturbance in cerebral function. However, the reasons for age-related expression in children are not understood. Neither are the mechanisms whereby an epileptic encephalopathy originates. Several new drugs have been recently introduced but have provided limited therapeutic benefits. However, treatment and quality of life have improved because the syndrome-specific efficacy profile of drugs is better known, and there is heightened awareness that compounds with severe cognitive side-effects and heavy polytherapies should be avoided. Epilepsy surgery is an important option for a few well-selected individuals, but should be considered with great caution when there is no apparent underlying brain lesion. PMID- 16473128 TI - Stigma and global health: developing a research agenda. PMID- 16473129 TI - Stigma and its public health implications. PMID- 16473130 TI - Stigma and the law. PMID- 16473131 TI - Stigma, race, and disease in 20th century America. PMID- 16473132 TI - The many stigmas of mental illness. PMID- 16473133 TI - Obstetric fistula and stigma. PMID- 16473134 TI - Stigma interventions and research for international health. PMID- 16473135 TI - Stigma and global health: looking forward. PMID- 16473136 TI - Occam can be wrong: a young man with lumbar pain and acute weakness of the legs. PMID- 16473137 TI - Sleeping sickness: PEX and drugs. AB - Finding new ways in the treatment of fatal parasitic diseases like the human sleeping sickness is a major challenge of biomedical research. The growing body of knowledge about the biogenesis of the glycosome, a peroxisome-related organelle of trypanosomes, might allow defining novel targets for drug development. PMID- 16473138 TI - The telomerase inhibitor telomestatin induces telomere shortening and cell death in Arabidopsis. AB - The cellular response to telomere dysfunction in plants was investigated with the use of telomestatin, an inhibitor of human telomerase activity. Telomestatin bound to plant telomeric repeat sequence, and inhibited telomerase activity in suspension-cultured cells of Arabidopsis thaliana and Oryza sativa (rice) in a dose-dependent manner. The inhibitor did not affect transcript level of the TERT gene, which encodes the catalytic subunit of telomerase, in the plant cells. Inhibition of telomerase activity by telomestatin resulted in rapid shortening of telomeres and the induction of cell death by an apoptosis-like mechanism in Arabidopsis cells. These results suggest that telomerase contributes to the survival of proliferating plant cells by maintaining telomere length, and that telomere erosion triggers cell death. PMID- 16473139 TI - Expression levels and subcellular localization of Bcy1p in Candida albicans mutant strains devoid of one BCY1 allele results in a defective morphogenetic behavior. AB - We investigated expression, functionality and subcellular localization of C. albicans Bcy1p, the PKA regulatory subunit, in mutant strains having one BCY1 allele fused to a green fluorescent protein (GFP). DE-52 column chromatography of soluble extracts of yeast cells from strains bearing one BCY1 allele (fused or not to GFP) showed co-elution of Bcy1p and Bcy1p-GFP with phosphotransferase activity, suggesting that interaction between regulatory and catalytic subunits was not impaired by the GFP tag. Subcellular localization of Bcy1p-GFP supports our previous hypothesis on the nuclear localization of the regulatory subunit, which can thus tether the PKA catalytic subunit to the nucleus. Protein modeling of CaBcy1p, showed that the fusion of the GFP tag to Bcy1p C-terminus did not significantly disturb its proper folding. Bcy1p levels in mutant strains having one or both BCY1 alleles, led us to establish a direct correlation between the amount of protein and the number of alleles, indicating that deletion of one BCY1 allele is not fully compensated by overexpression of the other. The morphogenetic behavior of several C. albicans mutant strains bearing one or both BCY1 alleles, in a wild-type and in a TPK2 null genetic background, was assessed in N acetylglucosamine (GlcNAc) liquid medium at 37 degrees C. Strains with one BCY1 allele tagged or not, behaved similarly, displaying pseudohyphae and true hyphae. In contrast, hyphal morphology was almost exclusive in strains having both BCY1 alleles, irrespective of the GFP insertion. It can be inferred that a tight regulation of PKA activity is needed for hyphal growth. PMID- 16473140 TI - Molecular alterations of cells resistant to platinum drugs: role of PKCalpha. AB - Development of resistance to platinum compounds may involve not only overexpression of defence mechanisms but also alterations in cellular response to the drug-induced genotoxic stress. To investigate the cellular bases of response to platinum compounds, we examined the profile of gene expression of ovarian carcinoma cells exhibiting sensitivity (A2780) or resistance (A2780/BBR3464) to platinum compounds. Using display PCR, we found that acquisition of resistance to the multinuclear platinum complex BBR3464 was associated with modulation of several transcripts, including up-regulation of the major substrate of protein kinase C (PKC), the myristoylated alanine-rich C kinase substrate (MARCKS). This feature was associated with PKCalpha down-regulation. To explore the role of PKCalpha in cellular sensitivity to platinum compounds, resistant cells were transfected with a PKCalpha-containing vector. PKCalpha-overexpressing resistant cells exhibited a decrease in sensitivity to cisplatin, whereas no significant change in sensitivity to BBR3464 was observed. A number of approaches designed to modulate the function or expression of PKCalpha support that the isoenzyme may play a role in determining resistance only to cisplatin but not to BBR3464, which is known to activate a different pathway of cell response. In conclusion, in spite of PKCalpha down-regulation in our model, its regulatory function was not apparently implicated in the development of resistance to platinum compounds and the present results do not support a general role of PKCalpha as a determinant of the resistance status. PMID- 16473141 TI - Whet the volunteer appetite. PMID- 16473147 TI - CPR in the box. PMID- 16473148 TI - CPR in the box. PMID- 16473149 TI - CPR in the box. PMID- 16473150 TI - It's not what you say, but what you write. PMID- 16473151 TI - What makes it work? Helping people understand their cots. PMID- 16473152 TI - AAA calls on Congress for Medicare relief. PMID- 16473153 TI - Planning ahead. Albuterol & the potential for hypokalemia-induced ventricular tachycardia in prehospital care. PMID- 16473155 TI - 7 steps to a good SOP. Why are we setting up our employees for failure? PMID- 16473156 TI - The vague OD. The symptomatic patient with an unknown exposure. PMID- 16473157 TI - Get to know Bryan Bledsoe, DO, FACEP. PMID- 16473158 TI - Have you hugged your physician advisor today? PMID- 16473161 TI - Recommended clinical competencies for initiating a program in endovascular repair of the thoracic aorta. PMID- 16473159 TI - Crew & patient safety receive high priority. PMID- 16473162 TI - Current Procedural Terminology (CPT) coding for descending thoracic aorta endovascular repair. PMID- 16473163 TI - Branched and fenestrated stent grafts for endovascular repair of thoracic aortic aneurysms. PMID- 16473164 TI - US multicenter trials of endoprostheses for the endovascular treatment of descending thoracic aneurysms. PMID- 16473165 TI - Worldwide survey of thoracic endografts: practical clinical application. PMID- 16473166 TI - Endovascular treatment of traumatic thoracic aortic injury--should this be the new standard of treatment? PMID- 16473167 TI - Starting a program for endovascular thoracic procedures: challenges and solutions. PMID- 16473168 TI - Aortic dissection: perspectives in the era of stent-graft repair. PMID- 16473169 TI - Emergent stent-graft treatment for rupture. PMID- 16473170 TI - Endovascular management of thoracic aortic aneurysms. Preoperative imaging and device sizing. PMID- 16473171 TI - Intraprocedural imaging: thoracic aortography techniques, intravascular ultrasound, and special equipment. PMID- 16473172 TI - Current results of open surgical repair of descending thoracic aortic aneurysms. PMID- 16473173 TI - Techniques for large sheath insertion during endovascular thoracic aortic aneurysm repair. PMID- 16473174 TI - Percutaneous techniques for aneurysm repair. PMID- 16473175 TI - Subclavian artery transposition and bypass techniques for use with endoluminal repair of acute and chronic thoracic aortic pathology. PMID- 16473176 TI - Managing proximal arch vessels. PMID- 16473177 TI - Visceral vessel relocation techniques. PMID- 16473178 TI - Complications of thoracic aortic endografts: spinal cord ischemia and stroke. PMID- 16473179 TI - Postoperative imaging surveillance and endoleak management after endovascular repair of thoracic aortic aneurysms. PMID- 16473180 TI - Late complications of thoracic endografts. PMID- 16473181 TI - A l'oeil malade la lumiere nuit. PMID- 16473183 TI - Intraoperative floppy iris syndrome associated with tamsulosin. PMID- 16473184 TI - Antipsychotic agent as an etiologic agent of IFIS. PMID- 16473185 TI - Intraoperative floppy iris syndrome associated with tamsulosin. PMID- 16473188 TI - Phaco rolling technique. PMID- 16473190 TI - Alpha antagonists in cataract surgery. PMID- 16473191 TI - How dark is "dark"? Experimental methods in clinical research. PMID- 16473193 TI - Comparative study of first-day postoperative cataract review methods. PMID- 16473194 TI - Autorefraction versus subjective refraction. PMID- 16473197 TI - Amblyopic adult eyes after LASIK. PMID- 16473199 TI - Phakic refractive lenses and vitreous dislocation. PMID- 16473200 TI - Pupil response to tropicamide following LASIK. PMID- 16473201 TI - Mini-nuc technique. PMID- 16473202 TI - Consultation section: cataract surgical problem. PMID- 16473213 TI - Two-step procedure to enlarge small optical zones after photorefractive keratectomy for high myopia. AB - We describe a method for the visual rehabilitation of patients with small optical zones and related complaints after photorefractive keratectomy (PRK) and laser in situ keratomileusis for high myopia. In many of these cases from the early 1990s, low central corneal thickness in combination with residual myopia did not allow for enlargement of the small optical zone by a topography-guided treatment in the first instance. We therefore perform, as a first step, a clear lens exchange aiming at hyperopia. This enables us to perform, as a second step, a topography guided customized PRK with marked enlargement of the optical zone. Such 2-step procedures may be of great benefit to patients with small optical zones and low central corneal thickness. PMID- 16473214 TI - Modification of the high-speed vitrectomy system TSV 25 for pediatric cataract surgery. AB - Modifications of a transconjunctival sutureless vitrectomy system (TSV 25, Bausch & Lomb Surgical) for removing cataracts and performing poster capsulotomy anterior vitrectomy in children are reported. PMID- 16473215 TI - Limbal relaxing incisions versus on-axis incisions to reduce corneal astigmatism at the time of cataract surgery. AB - PURPOSE: To compare limbal relaxing incisions (LRIs) with placement of the corneal cataract incision on the steepest keratometric axis for the reduction of preexisting corneal astigmatism at the time of cataract surgery. SETTING: The Queen Elizabeth Hospital, Adelaide, South Australia, Australia. METHODS: In a prospective single center study, patients having 1.5 diopters (D) or more of keratometric astigmatism were randomly assigned to 2 surgical techniques: on-axis incisions (OAIs) consisting of a single clear corneal cataract incision centered on the steepest corneal meridian or LRIs consisting of 2 arcuate incisions straddling the steepest corneal meridian and a temporal clear corneal incision. Vector analysis of the target axis flattening effect was used to assess the efficacy of treatment. RESULTS: Seventy-one eyes of 71 patients were evaluated, 33 in the OAI group and 38 in the LRI group. Six weeks postoperatively, the flattening effect was 0.41 D (median and interquartile range 0.15 to 0.78 D) in the OAI group and 1.21 D (range 0.43 to 2.25 D) in the LRI group (P = .002). After 6 months, the flattening effect was 0.35 D (range 0.00 to 0.96 D) and 1.10 D (range 0.25 to 1.79 D), respectively (P = .004). CONCLUSION: The amount of astigmatism reduction achieved at the intended meridian was significantly more favorable with the LRI technique, which remained consistent throughout the follow up period. PMID- 16473216 TI - Secondary Artisan-Verysise aphakic lens implantation. AB - PURPOSE: To evaluate efficacy, predictability and safety of Artisan-Verysise intraocular lens (IOL) secondary implantation for aphakia correction. SETTING: Instituto de Microcirugia Ocular, and Autonoma University of Barcelona, Barcelona, Spain. METHODS: Uncorrected visual acuity, best spectacle-corrected visual acuity (BSCVA), manifest refraction, endothelial cell count, and clinical complications were evaluated. Sixteen consecutive eyes of 14 patients with aphakia were submitted to surgery. Postoperative examinations were done at 6 weeks, 6 months, 1 year, and every year for at least 3 years. An iris-supported Artisan-Verysise IOL was implanted for aphakia correction. RESULTS: Thirty-six months after Artisan-Verysise lens implantation, BSCVA was 20/40 or better in 6 eyes (37.5%). Preoperatively, 5 eyes had the same BSCVA (31.25%). Mean postoperative spherical equivalent (SE) was 0.46 diopter (D). Mean endothelial cell loss was 10.9% 36 months postoperatively. The cell loss occurred predominantly during the first year (7.78%). Cystoid macular edema was observed in 2 cases, 1 of them associated with chronic unresponsive low intraocular pressure. No other serious complications were observed. CONCLUSION: Artisan Verysise IOL implantation seems a safe, predictable, and effective option for aphakic eyes without capsule support. PMID- 16473217 TI - Influence of pupil and optical zone diameter on higher-order aberrations after wavefront-guided myopic LASIK. AB - PURPOSE: To investigate the influence of pupil and optical zone (OZ) diameter on higher-order aberrations (HOAs) after myopic wavefront-guided laser in situ keratomileusis (LASIK). METHODS: Twenty-seven myopic eyes of 19 patients were included. The mean preoperative spherical equivalent was -6.86 diopters (D) +/- 1.24 (SD) (range -4.25 to -9.5 D); the mean planned OZ diameter was 6.26 +/- 0.45 mm (range 5.7 to 7.1 mm). All patients had uneventful wavefront-guided LASIK (Zyoptix version 3.1, Bausch & Lomb) and an uncomplicated follow-up of 12 months. Wavefront measurements were performed with a Hartmann-Shack sensor in maximum mydriasis preoperatively and 12 months after LASIK. Wavefront errors were computed for pupil diameters (PDs) of 3.0, 3.5, 4.0, 5.0, 6.0, and 7.0 mm for the individual OZ diameter and for the individual mydriatic PD (7.93 +/- 0.46 mm). The impact of the relationship between pupil diameter and OZ diameter (fractional clearance [FC]) on HOA was described and quantified using curvilinear regression with a 4th-order polynomial fit. RESULTS: There was a reproducible relationship between FC and the amount of induced HOA. The change in HOA root mean square and primary spherical aberration (Z(4)0) was significantly correlated with FC. If the OZ was 16.5% larger than the pupil (FC = 1.17), only half the amount of HOA was expected to be induced than if the OZ equaled the pupil. In contrast, an OZ that was 9% smaller than the pupil (FC = 0.91) resulted in an HOA induction 50% higher than at FC = 1. CONCLUSION: The OZ zone to pupil ratio (fractional clearance) had a significant impact on HOA induction after wavefront-guided LASIK. PMID- 16473218 TI - Laser in situ keratomileusis in patients with corneal guttata and family history of Fuchs' endothelial dystrophy. AB - PURPOSE: To report 1-year results of laser in situ keratomileusis (LASIK) in 7 eyes with corneal endothelial guttata and a family history of Fuchs' endothelial dystrophy. SETTING: John Moran Eye Center, University of Utah, Salt Lake City, Utah, USA. METHODS: A retrospective chart review was performed of 4 patients (7 eyes) who had trace to 1+endothelial guttata and a family history of Fuchs dystrophy and then had uneventful LASIK for the correction of myopia and myopic astigmatism. Preoperative and postoperative measurements included uncorrected visual acuity (UCVA), best spectacle-corrected visual acuity (BSCVA), corneal pachymetry, endothelial cell density (ECD), manifest refraction, and spherical equivalent. The changes in ECD, pachymetry, and spherical equivalent after LASIK were subjected to statistical analysis using a paired Student t test to determine significance. RESULTS: Transient corneal edema was noted in the early postoperative period in 3 eyes of 2 patients. At 1 year, 6 of the 7 (86%) eyes had lost > or =2 lines of BSCVA. A statistically significant decrease in ECD of 12.4% +/- 2.7% was observed at 1 year compared with baseline (P < .001). An increase in corneal thickness (P = .006) and a statistically significant myopic shift in spherical equivalent (P = .017) was also noted at 1 year compared with 3 months. CONCLUSIONS: Patients with mild corneal guttata and a family history of Fuchs' dystrophy are prone to transient corneal edema, loss of BSCVA, endothelial cell loss, and myopic regression after uneventful LASIK for correction of myopia and myopic astigmatism. PMID- 16473219 TI - Contact lens overrefraction variability in corneal power estimation after refractive surgery. AB - PURPOSE: To evaluate the accuracy and precision of the contact lens overrefraction (CLO) method in determining corneal refractive power in post refractive-surgery eyes. SETTING: Refractive Surgery Service and Contact Lens Service, University of Illinois, Chicago, Illinois, USA. METHODS: Fourteen eyes of 7 subjects who had a single myopic laser in situ keratomileusis procedure within 12 months with refractive stability were included in this prospective case series. The CLO method was compared with the historical method of predicting the corneal power using 4 different lens fitting strategies and 3 refractive pupil scan sizes (3 mm, 5 mm, and total pupil). Rigid lenses included 3 9.0 mm overall diameter lenses fit flat, steep, and an average of the 2, and a 15.0 mm diameter lens steep fit. Cycloplegic CLO was performed using the autorefractor function of the Nidek OPD-Scan ARK-10000. Results with each strategy were compared with the corneal power estimated with the historical method. The bias (mean of the difference), 95% limits of agreement, and difference versus mean plots for each strategy are presented. RESULTS: In each subject, the CLO-estimated corneal power varied based on lens fit. On average, the bias between CLO and historical methods ranged from -0.38 to +2.42 diopters (D) and was significantly different from 0 in all but 3 strategies. Substantial variability in precision existed between fitting strategies, with the range of the 95% limits of agreement approximating 0.50 D in 2 strategies and 2.59 D in the worst-case scenario. The least precise fitting strategy was use of flat-fitting 9.0 mm diameter lenses. CONCLUSIONS: The accuracy and precision of the CLO method of estimating corneal power in post refractive-surgery eyes was highly variable on the basis of how rigid lense were fit. One of the most commonly used fitting strategies in clinical practice--flat fitting a 9.0 diameter lens-resulted in the poorest accuracy and precision. Results also suggest use of large-diameter lenses may improve outcomes. PMID- 16473220 TI - Photorefractive keratectomy with intraoperative mitomycin-C application. AB - PURPOSE: To evaluate the efficacy and safety of photorefractive keratectomy (PRK) with intraoperative application of mitomycin-C (MMC). SETTING: Yonsei Eye Center, Seoul, South Korea. METHODS: This retrospective noncomparative case series included 536 patients (1011 eyes) who had had PRK with intraoperative application of MMC using the Nidek EC-5000 excimer laser. Preoperative and postoperative best spectacle-corrected and uncorrected visual acuities, spherical equivalent (SE) refraction, corneal haze graded by slitlamp biomicroscopy, and endothelial cell density measured by specular microscopy were evaluated. RESULTS: The mean preoperative SE was -7.82 diopters (D) +/- 2.64 (SD); 72% of eyes (732) were more than -6.00 D, and 28% (287) were more than -9.00 D. The mean follow-up was 13 months (range 6 to 27 months). Six months postoperatively, the mean postoperative SE was -0.14 +/- 0.62 D; 86% were within +/-0.50 D and 93% were within +/-1.00 D of desired refraction. Eighty-six percent had 20/20 or better visual acuity, and 98% were 20/40 or better. Regression of more than 1.00 D occurred in 78 eyes (7.6%), and it was more common in eyes with a preoperative SE of -9.00 D or worse (18%). Haze occurred in 32 eyes (3.17%), but in most cases it was limited to grade 1. Grades 2 and 3 haze occurred in 3 eyes and 2 eyes, respectively. The postoperative endothelial cell density measured by specular microscopy did not show a significant difference from preoperative measurements. Delayed epithelial healing was observed in 2 eyes. CONCLUSION: Photorefractive keratectomy with intraoperative application of MMC was a safe procedure that produced excellent visual outcomes with few complications. PMID- 16473221 TI - Laser-assisted subepithelial keratectomy for dry eye associated with soft contact lenses. AB - PURPOSE: To evaluate the efficacy and safety of laser-assisted subepithelial keratectomy (LASEK) in myopic patients with preoperative dry eye associated with extended soft contact lens use. SETTING: Department of Ophthalmology, Taipei Veterans General Hospital, Taipei, Taiwan. METHODS: This nonrandomized prospective interventional case series study comprised 9 patients (18 eyes) with dry eye and keratoconjunctivitis associated with long-term complicated soft contact lens use. Prior to LASEK surgery and for 1 year following surgery, measurements and comparisons were made of uncorrected visual acuity, best corrected visual acuity (BCVA), manifest refraction, symptoms, tear function, ocular surface abnormality, and corneal sensitivity. RESULTS: No significant differences (P > .05) in the mean results of the Schirmer test with anesthesia and tear breakup times before and after LASEK were evident. Corneal sensitivity recovered within 1 month after LASEK. No complications occurred during or after LASEK. Subepithelial haze and superficial punctate epithelial defects occurred in 1 eye, representing a loss of 1 Snellen line of BCVA. CONCLUSION: Laser-assisted subepithelial keratomileusis in dry eyes and keratoconjunctivitis associated with complicated soft contact lens wearing was safe and efficacious. PMID- 16473222 TI - Enhancement outcomes after photorefractive keratectomy and laser in situ keratomileusis using topographically guided excimer laser photoablation. AB - PURPOSE: To evaluate the efficacy and safety of topographically guided excimer laser photoablation to retreat unsuccessful myopic and hyperopic photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK). SETTING: Eye Clinic, San Salvatore Hospital, University of L'Aquila, L'Aquila, Italy. METHODS: At least 3 months after primary PRK (Group A) or primary LASIK (Group B), 48 eyes of 42 patients were submitted to PRK or LASIK enhancements. The eyes were treated with an excimer laser linked to a computerized videokeratography unit with a topographically supported customized ablation workstation. RESULTS: The mean follow-up was 27.8 months +/- 8.2 (SD). In Group A, the uncorrected visual acuity (UCVA) changed from 0.5 +/- 0.7 logarithm of the minimum angle of resolution (logMAR) (range 20/600 to 20/200) to 0.1 +/- 0.7 logMAR (range 20/60 to 20/20); the mean best spectacle-corrected visual acuity (BSCVA) changed from 0.1 +/- 0.7 logMAR (range 20/50 to 20/20) to 0 +/- 0.7 logMAR (range 20/50 to 20/20) after the enhancement. In Group B, the UCVA changed from 0.7 +/- 0.8 logMAR (range 20/600 to 20/40) to 0.1 +/- 0.7 logMAR (range 20/40 to 20/20); the mean BSCVA improved from 0.2 +/- 0.8 logMAR (range 20/30 to 20/20) to 0 +/- 1.3 logMAR (range 20/25 to 20/20) after surgery. CONCLUSIONS: The enhancements using topographically guided excimer laser photoablation with a topographically supported customized ablation method resulted in satisfactory and stable visual outcome with good safety and efficacy after unsuccessful PRK and LASIK. PMID- 16473223 TI - Sensitivity of the National Eye Institute Refractive Error Quality of Life instrument to refractive surgery outcomes. AB - PURPOSE: To evaluate scores from the National Eye Institute Refractive Error Quality of Life instrument (NEI-RQL) relative to refractive surgery outcomes in myopic patients. SETTING: Large institutional ophthalmic practices. METHODS: Patients seeking refractive surgery (n = 59) self-administered the NEI-RQL before and 6 months after surgery; the comparison group of myopes and emmetropes (n = 72) not seeking refractive surgery also completed the NEI-RQL. Myopic patients seeking refractive surgery had bilateral, same-day laser in situ keratomileusis (LASIK) treatment. Regression analyses were conducted to compare survey scores in the LASIK treatment group before surgery with those in the myopic sample who were not seeking refractive surgery and to compare the survey scores in the post-LASIK treatment group (surgical emmetropes) to those in the naturally emmetropic group. Logistic regression was used to determine NEI-RQL subscales that were related to myopic patients seeking refractive surgery and myopic patients not seeking refractive surgery. RESULTS: Myopes seeking LASIK reported greater expectations for vision improvement (P < .0001), greater activity limitations (P < .0001), suboptimal correction (P = .04), worse appearance (P < .0001), and lower satisfaction (P < .0001) than myopes not seeking refractive surgery. Logistic regression showed that worse self-perceived appearance (odds ratio 20.71), more dissatisfaction with current correction (odds ratio 8.28), and higher expectations for "optimal" correction (odds ratio 6.41) were independently associated with myopes seeking LASIK, while better perceived "clarity of vision" (odds ratio = 0.20) was associated with myopes not seeking surgery. Overall, surgical emmetropes were similar to natural emmetropes in terms of refractive error-specific quality of life (clarity of vision, near vision, far vision, diurnal fluctuations) and clinical measures of visual function, yet they reported significantly better appearance (P = .02) and greater satisfaction (P < .0001) than natural emmetropes. CONCLUSION: The NEI-RQL demonstrated vision-related quality-of-life differences when comparing patient samples associated with refractive surgery. PMID- 16473225 TI - AcrySof Natural SN60AT versus AcrySof SA60AT intraocular lens in patients with color vision defects. AB - PURPOSE: To determine whether implantation of the AcrySof Natural intraocular lens (IOL) worsened the severity of existing color deficit in congenital partial red-green color deficient individuals (CPRG). METHODS: A prospective controlled randomized double-masked analysis of 30 consecutive patients with CPRG defect and bilateral cataracts received a Natural IOL (test group) in 1 eye and a single piece AcrySof IOL (control group) in the other eye. Patients were tested unilaterally to detect CPRG defect using Ishihara pseudoisochromatic plates and the Farnsworth D-15 test. Plates 1 to 21 measured the Ishihara error score; plates 22 to 25 indicated severity of defect based on clarity of both numerals as partial mild/moderate (both visible), partial severe defect (only 1 visible). The D-15 test is based on number of diametrical crossings on the circular diagram; severity is graded as mild (1 crossing), moderate (2 crossings), or severe (>2 crossings). Tests were performed before and after IOL implantation at 1, 3, and 6 months. At mean follow-up of 6.13 months +/- 1.2 (SD), analysis of variance test judged the difference in error scores and cross tabulation represented change in number of diametrical crossings. RESULTS: The mean age was 62.3 +/- 8.5 years. All patients were men. Before IOL implantation, all patients had moderate CPRG defect on both tests. The Ishihara error score in the test and control groups did not reveal statistically significant differences (P = .505 and P = .119, respectively). With D-15, none of the patients in the test or control group showed >2 crossings. CONCLUSION: The implantation of AcrySof Natural IOL did not worsen the preexisting severity of color defect in CPRG individuals. PMID- 16473224 TI - Clinical results of the blue-light filtering AcrySof Natural foldable acrylic intraocular lens. AB - PURPOSE: To verify the safety and effectiveness of the new AcrySof Natural (Alcon Laboratories, Inc.) blue-light filtering intraocular lens (IOL), which was designed to achieve a light-transmission spectrum similar to that of the natural human crystalline lens. SETTING: Multicenter U.S. clinical trial. METHODS: In a prospective randomized patient-masked multicenter study, 150 patients received the AcrySof Natural IOL and 147 patients received the AcrySof single-piece IOL as a control. Patients with bilateral age-related cataracts who were willing and able to wait at least 30 days between cataract procedures and had verified normal preoperative color vision were eligible for the study. Standardized surgery included a 4.0 to 5.0 mm capsulorhexis and phacoemulsification. All lenses were inserted in the capsular bag, with verification of in-the-bag placement of both haptics. In all bilateral implantation cases, the same model IOL was used in each eye. Postoperatively, contrast sensitivity and color perception were measured up to 180 days and up to 1 year (for visual acuity) after implantation. RESULTS: No statistically significant differences were discovered between the 2 patient groups in visual acuity, contrast sensitivity evaluated under mesopic and photopic conditions, or the number of patients who passed the Farnsworth D-15 color perception test. There were no lens-related adverse events in either group. CONCLUSIONS: The blue-light filtering AcrySof Natural IOL was equivalent to the conventional AcrySof lens in terms of postoperative visual performance. Additional long-term clinical studies should show whether the IOL actually provides the theoretical benefits to retinal health. PMID- 16473226 TI - Results of cataract extraction after implantable contact lens removal. AB - PURPOSE: To evaluate the visual results following insertion of implantable contact lenses (ICLs) in ametropic eyes and the development of subcapsular opacification with visual loss and to examine the anterior capsule, including the subcapsular tissue alteration, by light microscopy. SETTING: Department of Ophthalmology, Schlosspark-Klinik, affiliated hospital of the Charite Berlin, Humbold University, Berlin, Germany. METHODS: A prospective noncomparative interventional case series of anterior subcapsular cataracts in 9 of 127 (7.1%) patient eyes receiving ICLs to correct myopia and hyperopia was studied. The cataracts were phacoemulsified due to visual loss, and an intraocular lens (IOL) was implanted in the bag. After capsulorhexis, the anterior capsule was withdrawn for light microscopy examination. Visual acuity in each eye was measured before and after ICL implantation and before and after cataract extraction. The age range of cataract patients was 39 to 53 years. RESULTS: Implantable contact lens removal and phacoemulsification with IOL implantation for emmetropia resulted in an increased visual acuity compared to initial vision. Four of 28 hyperopic eyes (14.3%) developed subcapsular central opacification after ICL implantation, whereas 5 of 99 myopic patients (5.1%) developed opacifications. CONCLUSIONS: Patients should be informed prior to ICL implantation, there is a possibility of secondary subcapsular cataract formation and vision reduction. Although the posterior chamber inlay as well as the cataract can be removed and better acuity can be restored, a possible complication due to the ICL implantation cannot be avoided and the accommodation in young patients lost. PMID- 16473227 TI - Optical biometry of the anterior eye segment: interexaminer and intraexaminer reliability of ACMaster. AB - PURPOSE: To evaluate the interexaminer and intraexaminer reliability of corneal thickness, anterior chamber depth (ACD), and crystalline lens thickness measurements using a commercially available anterior segment optical biometry instrument (ACMaster, Carl Zeiss Meditec) based on partial coherence interferometry (PCI). SETTING: Medical University of Vienna, Vienna, Austria. METHODS: Interexaminer reliability and intraexaminer reliability were evaluated in 10 eyes of 10 young volunteers and 11 eyes of 11 cataract patients. The measurements of the interexaminer reliability were taken by 3 examiners. Corneal thickness, ACD, and lens thickness of the intraexaminer reliability were measured twice in all eyes by 1 examiner. To evaluate the effect of cycloplegia on the variability, the measurements were performed on 5 volunteers under cyclopentolate 1%. Measurements were performed using the prototype of the ACMaster based on PCI. RESULTS: The interexaminer/intraexaminer reliabilities were 99.9% for corneal thickness and ACD. The reliability of lens thickness could not be estimated because of a large number of missing values in the cataract patient group. The median interexaminer variability (SD) was 1.9 microm for corneal thickness, 7.5 microm for ACD, and 10.6 microm for lens thickness. The median intraexaminer variability (SD) was 1.6 microm for corneal thickness, 10.8 microm for ACD, and 8.7 microm for lens thickness. With cycloplegia, both the interexaminer variability and intraexaminer variability were smaller than without cycloplegia. CONCLUSIONS: Partial coherence interferometry measurements of anterior chamber distances (corneal thickness, ACD, lens thickness) using the prototype of ACMaster were highly reliable, allowing examiner-independent measurements. However, lens thickness measurements in cataract eyes were often difficult. PMID- 16473228 TI - Diffuse lamellar keratitis complicating laser in situ keratomileusis: post marketing surveillance of an emerging disease in British Columbia, Canada, 2000 2002. AB - PURPOSE: To describe a surveillance system and summarize data between January 2000 and December 2002 regarding diffuse lamellar keratitis (DLK), a complication of laser in situ keratomileusis (LASIK) surgery. SETTING: Community-based clinics in British Columbia, Canada, in which LASIK surgery is performed. METHODS: Monthly, all clinics in which LASIK is performed reported the number of LASIK procedures and nonnominal cases of DLK (by grade and onset date) to the British Columbia Centre for Disease Control. Diffuse lamellar keratitis outbreaks were investigated, and prevention and control measures were recommended. RESULTS: From 2000 to 2002, approximately 72,000 LASIK procedures were performed, with a mean DLK incidence rate of 0.67% (95% confidence interval, 0.61-0.73). The overall proportion of DLK cases attributed to outbreaks was 64%, decreasing from 72% in 2000 to 40% in 2003. CONCLUSIONS: An effective DLK surveillance program was implemented at all laser refractive clinics in British Columbia. Reported DLK incidence was 0.67 cases per 100 procedures, with 64% occurring in outbreaks. PMID- 16473229 TI - Comparative study of 2 anesthesia techniques for pediatric refractive surgery. AB - PURPOSE: To compare the effectiveness of 2 anesthesia techniques for pediatric refractive surgery, propofol/fentanyl and ketamine/midazolam. SETTING: Department of Ophthalmology, Magraby Eye & Ear Center, Muscat, Oman. METHODS: This prospective clinical study was of children ranging in age range from 3 to 12 years who were followed for myopic anisometropia with amblyopia after failure of conventional correction or patients with superficial stromal corneal opacities. The children were scheduled to have refractive surgery under general anesthesia using propofol/fentanyl (P/F group) or ketamine/midazolam (K/M group). All drugs were administered intravenously. RESULTS: Thirty patients were included in the study and were divided randomly into 2 equal groups. Both groups were comparable in age, weight, sex, duration of anesthesia, and duration of surgery. Time to complete recovery was significantly shorter in the P/F group. The hemodynamic changes observed in this study were not of clinical significance. Three patients in the P/F group showed arterial oxygen (O2) saturation of less than 90% and supplemental O2 was given by nasal cannula. The incidence of postoperative agitation and vomiting was significantly higher in the K/M group, while the need for jaw thrust was significantly higher in the P/F group. The difference between both groups in the ophthalmologist satisfaction score was not significant. CONCLUSIONS: Propofol offered some advantages for brief procedures outside the traditional operating rooms. It is short acting, and has a rapid offset resulting in shorter postoperative monitoring and a smoother recovery profile. It has greater potential for respiratory depression than ketamine. Hence, greater vigilance and experience with the pediatric airway is recommended with its use. PMID- 16473230 TI - Goodness-of-prediction of Zernike polynomial fitting to corneal surfaces. AB - PURPOSE: To determine the goodness-of-prediction of the fitting routine by measuring the difference between topographic corneal surfaces and their Zernike reconstructions as a function of polynomial order and optical zone size for various corneal conditions. SETTING: Corneal research laboratory in a university eye center. METHODS: Corneal topography maps (N = 253) were obtained from the Louisiana State University Eye Center. A variety of corneal conditions were used: normals; astigmatism; laser in situ keratomileusis, photorefractive keratoplasty (PRK), and radial keratotomy (RK) postoperative cases (myopic spherical corrections); keratoconus suspect; mild, moderate, and severe keratoconus; pellucid marginal degeneration; contact lens-induced corneal warpage; and penetrating keratoplasty. The root-mean-square (RMS) error of the goodness-of prediction of the Zernike representation of corneal surface elevation was extracted for 4, 6, and 10 mm optical zones, whereas Zernike radial orders were varied from 3 to 14 in 1-order steps. The mean +/- SEM of the RMS error was plotted as a function of Zernike order and compared with criteria for normal surface fits. RESULTS: Fitting accuracy improved as more Zernike terms were included, but some conditions showed significant errors (when compared with normal surfaces), even with many added terms. For a 6 mm optical zone, the normal cornea group had the lowest RMS error and did not require terms above the 4th order to achieve <0.25 microm RMS error. Astigmatism met the 0.25 microm threshold at the 5th order, whereas keratoconus suspect required 7 orders. Laser in situ keratomileusis and PRK met the 0.25 microm threshold at the 8th order, whereas RK required 10 orders. Contact lens-induced corneal warpage and mild keratoconus needed 12 orders to meet the 0.25 microm threshold, whereas pellucid marginal degeneration, moderate and severe keratoconus, and keratoplasty categories were not well fitted even at 14 orders. CONCLUSIONS: A 4th-order Zernike polynomial appeared reliable for modeling the normal cornea, but using a 4th-order fitting routine with an abnormal corneal surface caused a loss of fine detail shape information. As more Zernike terms were added, the accuracy of the fit improved, and the result approached the minimum error found with normal corneas. Unless sufficient higher-order Zernike terms are included when analyzing irregular surfaces, some diagnostic applications of Zernike coefficients may not be rigorous. This conclusion also suggests that wavefront shape analysis is similarly dependent on the number of orders used. Current surgical corrections based on normal-eye wavefronts may fail to capture all visually relevant aberrations in abnormal eyes, such as those having laser retreatments or experiencing corneal warpage from contact lens wear. A clinical goodness-of-fit or goodness-of-prediction index would indicate whether the number of terms in use has fully accounted for all of the visually significant aberrations present in the eye. PMID- 16473231 TI - Effective corneal refractive diameter as a function of the object tangent angle in visual space. AB - PURPOSE: To determine whether the currently accepted method of selecting a minimum optical zone diameter for laser refractive surgery that is equal to or slightly greater than the dark-adapted pupil diameter provides a sufficient diameter of corneal surface to focus light arising from objects in the paracentral and peripheral visual field. SETTING: Department of Ophthalmology and Visual Sciences, Texas Tech University Health Sciences Center, Lubbock, Texas, USA. METHODS: An optical model of the anterior segment was developed to calculate the effective corneal refractive diameter (ECRD), which is the diameter of the area of cornea that refracts all incident light rays arising from an object through the physical pupil (PP). This model incorporates the patient variables of central anterior chamber depth (ACD), central corneal curvature (K(c)), and the diameter of the apparent entrance pupil (EP). The model was expanded to incorporate distant objects off the line of sight (LOS), described by their angular displacement from the fixation object in visual space (the object tangent angle delta(ob)). Results were calculated for the 360 meridian degree visual field (ie, for all objects in visual space perceptually displaced from the fixation object by angle delta(ob)). The effect of the prolate nature of the cornea was also investigated. RESULTS: The ECRD expanded rapidly as a function of PP and delta(ob) but was minimally influenced by K(c). Beyond a critical object tangent angle delta(c), light rays striking the corneal vertex were not refracted through the PP, and the ECRD became an annular surface centered on the corneal vertex. The delta(c) was not a function of K, but increased as the PP increased and decreased as the ACD increased. The prolate nature of the cornea had little influence on the ECRD, even for very peripheral light rays. CONCLUSIONS: The ECRD expands rapidly when considering distant objects only slightly displaced from the LOS. A patient treated with an optical zone equal to or slightly greater than the dark-adapted pupil diameter may experience vision quality loss for paracentral and midperipheral objects even under conditions of ambient indoor lighting. PMID- 16473232 TI - Quantitative assessment of ophthalmic viscosurgical device retention using in vivo confocal microscopy. AB - PURPOSE: To develop and apply a new laboratory method for in vivo quantitative assessment of the retention of ophthalmic viscosurgical devices (OVDs) following phacoemulsification. SETTING: Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas, Texas, USA. METHODS: Studies of both eyes of New Zealand White rabbits were performed. Six OVDs were evaluated: Provisc and Healon (both sodium hyaluronate 1%), Healon5 (sodium hyaluronate 2.3%), Amvisc Plus (sodium hyaluronate 1.6%), Viscoat (sodium hyaluronate 3%-chondroitin sulfate 4%), and a new viscous-dispersive OVD, DisCoVisc (sodium hyaluronate 3% chondroitin sulfate 4%). The OVD was injected to fill the anterior chamber and a phacoemulsification needle inserted with the tip positioned just anterior to the lens capsule. Simulated phacoemulsification was performed for 1 minute using flow rates of 20, 40, and 60 mL/min; a vacuum level of 300 mm Hg; and ultrasound power of 60% using a Legacy phacoemulsification unit. The needle was removed, and silicone oil (1000 centistokes) was injected into the anterior chamber. The distance between the corneal endothelium and the OVD-silicone oil interface was measured using in vivo confocal microscopy through-focusing (CMTF). RESULTS: Significant differences in residual thickness were found between the OVDs tested. Specifically, the residual thickness of both DisCoVisc (mean 324.5 microm +/- 163.7 [SD]) and Viscoat (251.4 +/- 100.9 microm) was significantly greater than that of Provisc (9.5 +/- 16.7 microm), Healon (3.8 +/- 11.3 microm), Healon5 (0.6 +/- 2.4 microm), and Amvisc Plus (65.6 +/- 134.0 microm) (P < .05, Dunn test). Ophthalmic viscosurgical device retention was greatest with DisCoVisc; however, there was no statistically significant difference between DisCoVisc and Viscoat in residual thickness. The flow rate did not have a significant effect on the residual thickness (Friedman 2-way analysis of variance by ranks). CONCLUSIONS: Residual OVD thickness following simulated phacoemulsification could be quantitatively measured using in vivo CMTF. The results are consistent with human surgical experience in that the dispersive OVD (Viscoat) was better retained than the cohesive compounds. DisCoVisc, a new viscous-dispersive OVD, also showed retention compared with Viscoat under the experimental conditions. PMID- 16473233 TI - Evaluation of posterior capsule opacification using a new posterior view method in rabbits: single-piece acrylic versus 3-piece acrylic intraocular lens. AB - PURPOSE: To introduce a new procedure for evaluating posterior capsule opacification (PCO) in rabbit eyes and to perform a comparative study of the single-piece and 3-piece acrylic intraocular lenses (IOLs) on PCO using the new evaluation method. SETTING: Jinshikai Medical Foundation, Nishi Eye Hospital, Osaka, Japan. METHODS: A single-piece or 3-piece acrylic IOL was implanted in 1 eye and the other in the contralateral eye of 5 rabbits. Three weeks after surgery, PCO was scored by Evaluation of Posterior Capsule Opacification (EPCO) in posterior view. Before the posterior view was photographed, the anterior capsule was removed from the whole optic area to eliminate disturbing anterior capsule opacification (ACO) for the PCO evaluation. RESULTS: Posterior capsule opacification could be well observed and viewed in the posterior view so that it could be scored by EPCO without confusion with ACO. The mean PCO score of the single-piece and 3-piece acrylic IOLs was 3.12 +/- 0.19 and 2.41 +/- 0.70, respectively (P < .05 and P = .03, respectively). CONCLUSION: The removal of ACO allowed scoring of PCO by EPCO in rabbit eyes. The single-piece acrylic IOL showed significantly more PCO than the 3-piece acrylic IOL at least 3 weeks after surgery in rabbits. PMID- 16473234 TI - Lens epithelial cell ongrowth: comparison of 6 types of hydrophilic intraocular lens models. AB - PURPOSE: To compare the ongrowth of lens epithelial cells (LECs) on the anterior surface of 6 different hydrophilic intraocular lenses (IOLs). SETTING: Medical University of Vienna, Department of Ophthalmology and Optometry, Vienna, Austria. METHODS: Six models of hydrophilic IOLs were compared in this prospective study: Visionflex A-100 (Distra Softcryl), Rayner Centerflex 570H (Rayner), Collamer CC4204BF-IOL (Staar), Injectacryl F 3000 (Distra), Hydroview H60M (Bausch & Lomb), and MemoryLens (ORC). Postoperative biomicroscopic examinations were performed 1, 3, 7, 30, 90, 180, and 365 days after surgery. Lens epithelial cells in each quadrant of the anterior capsule-free lens surface were graded. The product with the highest density and the number of quadrants with this density were used to measure LEC ongrowth. RESULTS: The Hydroview and Visionflex IOLs showed significantly more LECs than the other IOLs starting on day 7 after surgery (P < .028). There was a statistically significant difference in LEC ongrowth on the Memory IOL compared with all other IOLs from day 30 onward (P < .001). The Rayner, Collamer and Injectacryl IOLs had the fewest LECs on the anterior surface compared with all other IOLs from day 7 until the final examination. CONCLUSIONS: The findings show that LEC ongrowth on the IOL surface is material dependent. The findings suggest that the material of the recently developed hydrophilic IOLs induces less LEC ongrowth than older models. PMID- 16473235 TI - Modulation transfer function and pupil size in multifocal and monofocal intraocular lenses in vitro. AB - PURPOSE: To investigate the relationship between pupil size and the modulation transfer function (MTF) of a multifocal intraocular lens (IOL) in vitro and to predict the visual effects in vivo. SETTING: Department of Ophthalmology and Visual Science, Kitasato University Graduate School of Medical Sciences, Kitasato, Japan. METHODS: A refractive multifocal IOL (Array SA-40N, Allergan) and a monofocal IOL (PhacoFlex SI-40NB, AMO) were evaluated using the OPAL Vector system and a model eye with a variable effective aperture. With effective pupil diameters of 2.1, 3.0, 3.4, 3.9, 4.6, 5.1, and 5.5 mm, the in-focus and defocus MTFs were measured in the multifocal and monofocal IOLs. RESULTS: With increases in effective pupil diameter, the far MTF progressively decreased at all spatial frequencies. In contrast, the near MTF began to increase at effective pupil diameter 2.1 mm, showed a peak at 3.4 mm, and decreased at diameters greater than 3.4 mm. The ratio of near MTF to far MTF showed an increase with larger effective pupil diameters and at lower spatial frequencies. CONCLUSIONS: With a zonal progressive multifocal IOL, the pupil size effected a trade-off between the far and near MTFs: The near MTF increased at the expense of the far MTF at large pupil sizes (effective pupil diameter >3.4 mm). To enhance near vision with a multifocal IOL, the desirable effective pupil diameter should be 3.4 mm or larger. PMID- 16473236 TI - First safety study of femtosecond laser photodisruption in animal lenses: tissue morphology and cataractogenesis. AB - PURPOSE: To determine through safety studies the tissue effects and potential cataractogenesis of laser modification of the crystalline lens (photophaco modulation). SETTING: Laser Zentrum Hannover, Hannover, Germany. METHODS: Six fresh porcine lenses and 6 living rabbit eyes (with the contralateral eye as a control) were radiated with a low-energy femtosecond laser to induce lens fiber disruption. After 3 months, the rabbit eyes were extracted and tested for light scatter and lens function and fixed for histology and ultrastructure. RESULTS: After laser treatment, all lenses displayed a tightly packed array of intralenticular bubbles, which resolved with time. In the porcine eyes, the bubbles coalesced unless spacing of 9 mum or greater was applied at an energy of 2 microJ. In the rabbit eyes, an energy of 1 microJ and spacing of 10 microm was chosen for transcorneal delivery, showing minimum bubble coalescence. After 3 months, the rabbit lenses showed good transparency, with only 1 rabbit having cataract formation unrelated to the laser. Laser scanning studies show essentially identical values for the back focal length and sharpness of focus (variability of back focal length). Ultrastructurally, the rabbit eyes showed a 0.5 microm electron dense border layer with adjacent normal lens architecture. CONCLUSIONS: Femtosecond laser photodisruption of the ocular lens yields a self limited lesion with bubbles that resolve with time. In living animal eyes, no cataract formation was found with no loss of lens function or induced light scatter after 3 months. These results suggest that use of a low-energy femtosecond laser might be safe when modifying the lens for presbyopia correction. PMID- 16473237 TI - Smoking and cataract: review of causal association. AB - Several risk factors for the development of cataract have been identified. This review evaluates epidemiologic literature that has examined tobacco smoking as a risk factor for cataract formation using established causality criteria. Twenty seven studies were included in this review. Evidence suggests that smoking has a 3-fold increase on the risk for incident nuclear cataract development. There was also evidence of dose response, temporal relationship, and reversibility of effect. There was limited evidence of an association between smoking and posterior subcapsular cataract, but little or no association with cortical cataract. Thus, the literature review indicated a strong association between smoking and the development of cataract, particularly nuclear cataract. The association fulfills the established criteria for causality. The association between smoking and other types of cataract is less distinct and requires further evaluation. PMID- 16473238 TI - ASCRS Binkhorst lecture 2004: the search for perfect vision: ophthalmology's Holy Grail? AB - Wavefront technology and wavefront-based vision correction are major advances but will not, in isolation, achieve the goal of perfect vision. Study of the animal kingdom teaches lessons of adaptation, leading to the conclusion that the human visual system is probably optimized for our ancestral hunter-gatherers, but today's visual demands have developed faster than the pace of evolution. Psychophysical studies of perception emphasize the importance of the visual processing system in the totality of visual function, with application today in areas such as multifocal intraocular lenses. The goal of "perfect vision" is more than optimized optics. A better understanding of visual function can be achieved only by a cooperative effort of ophthalmologists, vision scientists, and psychophysicists. PMID- 16473239 TI - Ophthalmology and vision science research: Part 3: avoiding writer's block- understanding the ABCs of a good research paper. AB - Completion of a scientific manuscript for submission to a peer-reviewed journal is a daunting task for clinicians and scientists early in their careers. In an ongoing series, this third article is the first of 2 related articles that deal with the basics of producing a high-quality research manuscript. Although ophthalmology and vision science are the principal focus of this series, the general concepts essential to producing a quality manuscript are applicable to diverse fields of research. This article highlights the exponential growth in the scientific literature over the past 40 years, considers why it is important to publish completed research, and discusses the necessity of identifying the key messages of the research, and their context, in relation to the published literature. The ethics of publishing biomedical research and scientific misconduct, such as duplicate publication or plagiarism, are outlined. To avoid later conflict, there is a critical need for coworkers to carefully address authorship order and inclusion early in the manuscript process. Internationally agreed guidelines are identified to guide this process. The importance of choosing the correct journal for a specific article and the nature of basic citation indices are discussed. The article concludes by elaborating and contrasting different scientific writing styles and emphasizing the considerable importance of developing a representative title and applying clarity and appropriate structure to the abstract. PMID- 16473240 TI - Complications arising from iris-fixated posterior chamber intraocular lenses. AB - Iris fixation of posterior chamber intraocular lenses (IOLs) is a technique that has been recommended for patients in whom a posterior chamber IOL is indicated but there is insufficient capsular support. We present 4 cases of iris-fixated posterior chamber IOLs with complications related to the procedure. Three patients developed postoperative slippage of the IOL, and 1 patient presented with iris erosion of the haptic leading to recurrent hemorrhage. Three cases were repaired by resuturing the same IOL using iris fixation, and in the last case IOL exchange with transscleral fixation of a posterior chamber IOL was performed. Final follow-up revealed stable IOLs in all cases. PMID- 16473241 TI - Origin of intraocular metallic foreign bodies during phacoemulsification. AB - This is the first report of a case of intraocular metallic foreign bodies retained after phacoemulsification in which the source was identified as silver emanating from the brazing of the irrigation tube as it entered the handpiece shell on its inner diameter. Although silver is considered to be inert in the eye and nonmagnetic, manufacturers should evaluate alternate methods of phacoemulsification handpiece design. PMID- 16473242 TI - Late spontaneous prolapse of a posterior chamber intraocular lens. AB - This is an unusual case of complete spontaneous anterior displacement of the optic of a posterior chamber intraocular lens (IOL) occurring nearly 12 years after its conventional insertion in the capsular bag. This occurred after the patient lowered her head. As the patient bent down to retrieve an object from underneath the bed, her vision became blurred on subsequent straightening and adoption of an upright standing posture. In this case, the haptic of the IOL likely disinserted at the time of pupil capture owing to possible previous damage during insertion. The optic spontaneously reverted to its intracapsular position. PMID- 16473243 TI - Imatinib as a cause of cystoid macular edema following uneventful phacoemulsification surgery. AB - Cystoid macular edema (CME) is an important cause of visual impairment following phacoemulsification surgery. Imatinib is a new drug that is being used as adjunctive therapy in the management of various malignancies. A reported ocular side effect of this drug is spontaneous retinal edema. We report the case of a patient on imatinib who developed significant premature CME following uneventful phacoemulsification surgery. This resolved soon after cessation of imatinib therapy, and there was an improvement in visual acuity. PMID- 16473244 TI - Tilt of single-piece hydrophobic intraocular lens due to folded haptic. PMID- 16473245 TI - In vitro adherence of Staphylococcus epidermidis, Serratia marcescens, and Pseudomonas aeruginosa to AcrySof intraocular lenses. PMID- 16473246 TI - Posterior dislocation of a crystalline lens associated with airbag deployment. PMID- 16473248 TI - Case management and the standards of practice. PMID- 16473249 TI - Flawed language or judges rule. PMID- 16473250 TI - Should I promote an internal candidate? PMID- 16473251 TI - Managing mental health disability in the workplace. AB - Jane is a 51-year-old customer-service representative for a utility company's customer-service call center. She has worked in this capacity for almost 10 years. She was considered an exemplary employee, received commendations at work, and was in line for a promotion to a supervisory position. Unexpectedly, she was assigned a new supervisor. Her performance began to decline, and her relationship with her supervisor became fraught with tension. A week later, Jane filed a grievance with her human resources department stating that her supervisor was verbally abusive, humiliated her in front of coworkers, and refused to accommodate her recently developing back pain. PMID- 16473252 TI - Are we being forced into isolation? AB - Do you ever feel a sense of being locked in? Has it been more than a year since you went out for lunch? Are e-mails your main source of keeping in touch with friends and colleagues? What are the issues for our self-inflicted isolation? What am I hearing and seeing? Case managers are saying they do not have time to meet with manufacturers or home care providers, and then the home care providers are upset that no one cares about what they have to offer. PMID- 16473253 TI - Mediation in health care: a collaborative journey. AB - In health care, the patient is the recipient of a service, another person provides that service, a third party attempts to standardize and regulate the service, and a fourth party creates invoices and expects a fifth party to pay for the service. One misstep in this game of dominoes and the ripple effect can cause adversity and conflict. In these situations there often is no wrong or right, only perceptions of situations, with perceived meanings and motives. Individuals' emotions and vision are often clouded in this web of perceptions, and mean spirited words may be exchanged. Once individuals have acted irrationally, they feel compelled to justify their actions, hence the escalation of conflict, each party vilifying the other and painting them as the enemy. These types of conflict abound in health care, consuming time, energy, and money. They divert attention from heath care's main focus-the patient-and affect everyone's ability to make wise and level-headed decisions. PMID- 16473254 TI - Self and balance amid chaos. AB - There I was, 18 months ago on my case management career fast track: working 12 hour days at the hospital, squeezing in time and energy to write those first articles and presentations, serving as president of my local Case Management Society of America chapter, occasionally waving to my husband, communicating by one-word e-mails with my son at college, and playing voice mail tag with family and friends. There was a nice, even flow to this work-though it resembled a car speeding along a highway yet never finding the exit. Know that feeling? PMID- 16473256 TI - Cardiometabolic risk assessment: an approach to the prevention of cardiovascular disease and diabetes mellitus. AB - An alarming increase in the development of cardiovascular disease (CVD) during the past 5 decades has led to intensive research on the epidemiology and pathogenesis of CVD, resulting in dramatic improvements in treatment. Today, there is an alarming increase in obesity and diabetes mellitus (DM), with a concomitant increase in diabetes-related complications, including CVD. Researchers have found that the risk of CVD becomes greater with increasing hyperglycemia and insulin resistance that occur in people long before the onset of clinical DM. Lifestyle modification with moderate weight loss has been shown to prevent or delay the onset of DM in patients who are at high risk for developing this disease. Unfortunately, the current guidelines for risk assessment provided by medical societies and national organizations are numerous, confusing, and inconsistent in their basic approach to identify specific risk factors for CVD and DM in patients. Extending routine systematic assessment from cardiovascular risk to cardiometabolic risk--that is, risk for developing CVD and/or DM--and increasing our understanding of the basic mechanisms that regulate energy balance and metabolic risk factors are needed to address this impending epidemic of DM. PMID- 16473257 TI - The endocannabinoid system: a new approach to control cardiovascular disease. AB - The endocannabinoid (EC) system consists of 2 types of G-protein-coupled cannabinoid receptors--cannabinoid type 1 (CB1) and cannabinoid type 2 (CB2)--and their natural ligands. The EC system plays a key role in the regulation of food intake and fat accumulation, as well as glucose and lipid metabolism. When overactivated, the EC system triggers dyslipidemia, thrombotic and inflammatory states, and insulin resistance. Blocking CB1 receptors centrally and peripherally in adipose tissue can help normalize an overactivated EC system. CB1 blockade helps regulate food intake and adipose tissue metabolism, contributing to improved insulin sensitivity and other features of the metabolic syndrome. Visceral adipose tissue is most closely associated with the metabolic syndrome, which is a constellation of conditions that place people at high risk for coronary artery disease. Targeting the EC system represents a new approach to treating visceral obesity and reducing cardiovascular risk factors. PMID- 16473259 TI - A constellation of complications: the metabolic syndrome. AB - The metabolic syndrome is a common cluster of risk factors for coronary heart disease and type 2 diabetes mellitus that includes obesity, elevated blood pressure, insulin resistance, and dyslipidemia. The diagnosis of metabolic syndrome itself appears to be an important risk factor for atherogenic cardiovascular disease and diabetes, and there is recent evidence that its components cluster, rather than occurring together by coincidence. A recent statement from the American Heart Association and the National Heart, Lung, and Blood Institute slightly modifies and clarifies the diagnostic criteria for the metabolic syndrome that are most widely used in the United States, along with giving practical guidance about management. Prompt therapeutic attention to the underlying risk factors--abdominal obesity, physical inactivity, and atherogenic/diabetogenic diet--is warranted for all patients with the metabolic syndrome, and drug therapy for specific metabolic risk factors should be considered for those at high or moderately high 10-year absolute risk of atherosclerotic cardiovascular disease. A new class of investigational drugs that block cannabinoid type 1 receptors have shown promise. This review also discusses issues that require additional research and new drugs that are considered promising for treatment of the metabolic syndrome itself. PMID- 16473258 TI - The metabolic basis of atherogenic dyslipidemia. AB - Atherogenic dyslipidemia is one of the major components of the metabolic syndrome, a complex cluster of several risk factors within a single patient that according to the National Cholesterol Education Program (NCEP) Adult Treatment Panel III includes at least 3 of the following: large waist circumference, elevated triglyceride levels, low levels of high-density lipoprotein cholesterol (HDL-C), hypertension, and elevated fasting glucose levels, which are directly related to the incidence of coronary heart disease. Atherogenic dyslipidemia clinically presents as elevated serum triglyceride levels, increased levels of small dense low-density lipoprotein (sdLDL) particles, and decreased levels of HDL-C. An important component of atherogenic dyslipidemia is central obesity, which is defined as increased waist circumference and has recently been identified as a chief predictor of the metabolic syndrome in certain patients. Another recent study found that both body mass index and waist circumference were highly predictive of eventual development of the metabolic syndrome. Because atherogenic dyslipidemia usually precedes the clinical manifestation of the metabolic syndrome, strategies to treat it are the focus of pharmacologic intervention. For example, the 3-hydroxy-3-methylglutaryl coenzyme-A reductase inhibitors, commonly known as statins, benefit hypercholesterolemic patients who have atherogenic dyslipidemia that is associated with the metabolic syndrome. Pioglitazone, an antidiabetic agent that acts primarily by decreasing insulin resistance, improves sensitivity to insulin in muscle and adipose tissue and inhibits hepatic gluconeogenesis. Pioglitazone improves glycemic control while reducing circulating insulin levels. The investigational agent, rimonabant--a centrally and peripherally acting, selective cannabinoid type-1 receptor blocker- is the first therapy developed for managing several cardiovascular risk factors at one time. Rimonabant has shown promise in attacking atherogenic dyslipidemia from several vantage points by affecting glucose, HDL-C, triglycerides, and waist circumference in patients who are prone to atherogenic dyslipidemia. PMID- 16473260 TI - Managing cardiovascular risk inpatients with metabolic syndrome. AB - The metabolic syndrome is a constellation of risk factors that contribute to the onset of type 2 diabetes mellitus and cardiovascular disease (CVD). CVD has been identified by the National Cholesterol Education Program (NCEP) as the primary clinical outcome of the metabolic syndrome. Although no algorithm is currently available for estimating the absolute risk of CVD for patients with the metabolic syndrome, screening for cardiovascular (CV) risk in these patients involves testing for lipoprotein abnormalities (namely, an analysis of specific low density lipoprotein particle numbers) and an assessment of various surrogate markers for subclinical coronary artery disease. Such screening can be used to help predict the development of CVD and thereby allow for effective interventions to help prevent coronary events. Strategies for reducing CV risk in patients with the metabolic syndrome are multifactorial. In addition to placing an emphasis on therapeutic lifestyle changes that increase levels of physical activity, dietary modification, and weight reduction, several pharmacologic therapies are available. One novel approach for managing CV risk in patients with the metabolic syndrome involves the inhibition of the endocannabinoid system, including the use of rimonabant. A review of CV risk factors in patients with the metabolic syndrome is beneficial for clinicians to apply in the care of their patients, along with a discussion about strategies for identifying at-risk patients and managing CVD risk for these patients. PMID- 16473261 TI - Pharmacotherapy to reduce visceral fat. AB - Cardiovascular disease (CVD) is the leading cause of death of men and women in the United States. The accumulation of visceral adipose tissue, as opposed to subcutaneous fat, increases the risk of developing CVD and metabolic disease. Because visceral adipose tissue uniquely contributes to the pathophysiology of CVD and insulin resistance, waist circumference is now being considered as a more useful marker of potential health risks associated with overweight and obesity than body mass index. Weight loss, particularly a reduction in waist circumference, improves insulin sensitivity, lipid profile, and serum adipocytokines, thereby reducing the risk of developing chronic disease and CVD. PMID- 16473262 TI - The metabolic syndrome, hyperlipidemia, and insulin resistance. AB - The metabolic syndrome is a cluster of cardiovascular risk factors associated with obesity. The defining components of the metabolic syndrome, according to the Third Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III), are elevated waist circumference, elevated triglycerides, reduced high-density lipoprotein cholesterol, elevated blood pressure, and elevated fasting glucose. The pathophysiologic basis of metabolic syndrome is complex and reflects several interrelated disturbances of glucose and lipid homeostasis. Metabolic syndrome is prevalent in the United States and developed nations, and patients with this disorder are at risk for type 2 diabetes mellitus and cardiovascular disease, underscoring the need for prompt patient identification and management. The first-line approach to control of metabolic syndrome is weight control and exercise. A wide range of pharmacologic interventions (eg, statins, antihypertensive drugs, insulin sensitizers, and thiazolidinediones) have been shown to be effective in controlling the individual components of metabolic syndrome. Obesity, which is a necessary component of metabolic syndrome, has reached epidemic proportions in the United States. Although lifestyle management or medications can control the underlying risk factors and most of the components of metabolic syndrome, long-term weight loss is difficult to achieve. Several promising pharmacologic interventions that may have an important role in the management of metabolic syndrome by treating adipose tissue-mediated metabolic disturbances are in the early stages of development. PMID- 16473265 TI - Draughtsmen, botanists and nature: constructing eighteenth-century botanical illustrations. AB - At first glance botanical illustrations of the eighteenth century might be interpreted as being naturalistic portraits of living plants. A more detailed investigation, however, reveals that the pictures were meant to communicate typical features of plant species in the way of a model. To this end, botanists of the period gave botanical draughtsmen specialist training; copying earlier examples and standardised motives from drawing books was a common part of this training. The practice of copying elements of previously published drawings and integrating them into new pictures was also widespread. However, only carefully selected elements were taken over, and even these were improved in terms of their correctness and appropriateness to the new context. This procedure was a strategy that eighteenth-century botanists used so that they would present an illustration that met their own requirements more satisfactorily than existing depictions. From this perspective, botanical illustrations can serve as historical sources on the working practices of eighteenth-century botanists and draughtsmen, which are usually not mentioned in textual sources. PMID- 16473266 TI - Darwin's artificial selection as an experiment. AB - Darwin used artificial selection (ASN) extensively and variedly in his theorizing. Darwin used ASN as an analogy to natural selection; he compared artificial to natural varieties, hereditary variation in nature to that in the breeding farm; and he also compared the overall effectiveness of the two processes. Most historians and philosophers of biology have argued that ASN worked as an analogical field in Darwin's theorizing. I will argue rather that this provides a limited and somewhat muddled view of Darwinian science. I say "limited" because I will show that Darwin also used ASN as a complex experimental field. And I say "muddled" because, if we concentrate on the analogical role exclusively, we conceive Darwinian science as rather disconnected from contemporary conceptions of "good science". I will argue that ASN should be conceived as a multifaceted experiment. As a traditional experiment, ASN established the efficacy of Darwin's preferred cause: natural selection. As a non traditional experiment, ASN disclosed the nature of a crucial element in Darwin's evolutionary mechanics: the nature of hereditary variation. Finally, I will argue that the experiment conception should help us make sense of Darwin's comments regarding the "monstrous" nature of domestic breeds traditionally considered to be problematic. PMID- 16473268 TI - What was Fisher's fundamental theorem of natural selection and what was it for? AB - Fisher's 'fundamental theorem of natural selection' is notoriously abstract, and, no less notoriously, many take it to be false. In this paper, I explicate the theorem, examine the role that it played in Fisher's general project for biology, and analyze why it was so very fundamental for Fisher. I defend and Lessard (1997) in the view that the theorem is in fact a true theorem if, as Fisher claimed, 'the terms employed' are 'used strictly as defined' (1930, p. 38). Finally, I explain the role that projects such as Fisher's play in the progress of scientific inquiry. PMID- 16473267 TI - Storytelling, statistics and hereditary thought: the narrative support of early statistics. AB - This paper's main contention is that some basically methodological developments in science which are apparently distant and unrelated can be seen as part of a sequential story. Focusing on general inferential and epistemological matters, the paper links occurrences separated by both in time and space, by formal and representational issues rather than social or disciplinary links. It focuses on a few limited aspects of several cognitive practices in medical and biological contexts separated by geography, disciplines and decades, but connected by long term transdisciplinary representational and inferential structures and constraints. The paper intends to show a given set of knowledge claims based on organizing statistically empirical data can be seen to have been underpinned by a previous, more familiar, and probably more natural, narrative handling of similar evidence. To achieve that this paper moves from medicine in France in the late eighteenth and early nineteenth century to the second half of the nineteenth century in England among gentleman naturalists, following its subject: the shift from narrative depiction of hereditary transmission of physical peculiarities to posterior statistical articulations of the same phenomena. Some early defenders of heredity as an important (if not the most important) causal presence in the understanding of life adopted singular narratives, in the form of case stories from medical and natural history traditions, to flesh out a special kind of causality peculiar to heredity. This work tries to reconstruct historically the rationale that drove the use of such narratives. It then shows that when this rationale was methodologically challenged, its basic narrative and probabilistic underpinings were transferred to the statistical quantificational tools that took their place. PMID- 16473269 TI - Rational versus anti-rational interpretations of science: an ape-language case study. AB - Robert Nola (2003) has argued that anti-rationalist interpretations of science fail to adequately explain the process of science, since objective reasons can be causal factors in belief formation. While I agree with Nola that objective reasons can be a cause of belief, in this paper I present a version of the strong programme in the sociology of knowledge, the Interests Thesis, and argue that the Interests Thesis provides a plausible explanation of an episode in the history of ape-language research. Specifically, I examine Terrace, Petitto, Sandess, & Bever (1979, 1980) illegitimate comparison of the signing of their chimpanzee, Nim, with data from human early childhood language development, and argue that Terrace et al.'s interests played a causal role in determining their sceptical beliefs concerning ape linguistic abilities. However, I go on to argue that Terrace et al.'s interests are not the only causal factors in determining their beliefs: objective reasons, associated with the institution of new methodologies, were also causally determinative of Terrace et al.'s sceptical beliefs. Consequently, I argue that belief formation in science is a multi-factorial affair wherein both interests and objective reasons have causal roles. I finish the paper with two conjectures concerning the proper locus of scientific rationality. PMID- 16473270 TI - Norms for emotions: biological functions and representational contents. AB - Normative standards are often applied to emotions. Are there normative standards that apply to emotions in virtue solely of facts about their nature? I will argue that the answer is no. The psychological, behavioural, and neurological evidence suggests that emotions are representational brain states with various kinds of biological functions. Facts about biological functions are not (and do not by themselves entail) normative facts. Hence, there are no normative standards that apply to emotions just in virtue of their having various kinds of biological functions. Moreover, the peculiar features of emotions make the view that representational content is essentially normative very implausible. Hence, the representational properties of emotions cannot be seen as entailing normative standards. The conclusion is that there are no normative standards that apply to emotions solely in virtue of their nature. PMID- 16473271 TI - How Darwinian reductionism refutes genetic determinism. AB - Genetic determinism labels the morally problematical claim that some socially significant traits, traits we care about, such as sexual orientation, gender roles, violence, alcoholism, mental illness, intelligence, are largely the results of the operation of genes and not much alterable by environment, learning or other human intervention. Genetic determinism does not require that genes literally fix these socially significant traits, but rather that they constrain them within narrow channels beyond human intervention. In this essay we analyze genetic determinism in light of what is now known about the inborn error of metabolism phenylketonuria (PKU), which has for so long been the poster child 'simple' argument in favor of some form of genetic determinism. We demonstrate that this case proves the exact opposite of what it has been proposed to support and provides a strong refutation of genetic determinism in all its guises. PMID- 16473272 TI - Anarchy, socialism and a Darwinian left. AB - In A Darwinian left Peter Singer aims to reconcile Darwinian theory with left wing politics, using evolutionary game theory and in particular a model proposed by Robert Axelrod, which shows that cooperation can be an evolutionarily successful strategy. In this paper I will show that whilst Axelrod's model can give support to a kind of left wing politics, it is not the kind that Singer himself envisages. In fact, it is shown that there are insurmountable problems for the idea of increasing Axelrodian cooperation within a welfare state. My surprising conclusion will be that a Darwinian left worthy of the name would be anarchistic. PMID- 16473273 TI - Reconstructing a disease: What essential features of the retinoic acid receptor fusion oncoproteins generate acute promyelocytic leukemia? AB - Acute promyelocytic leukemia (APL) is associated with rearrangement of the retinoic acid receptor alpha (RARalpha) gene leading to the formation of chimeric receptor proteins. In this issue of Cancer Cell, studies by Kwok et al. and Sternsdorf et al. indicate that the ability of the RARalpha oncoproteins to dimerize/multimerize is an essential feature required for the development of disease. Homodimerization allows RARalpha to bind to corepressors with increased affinity and the ability to bind to novel DNA sequences. However, artificial RARalpha dimers were weak oncogenes in vivo, indicating that the fusion partners confer additional properties to RARalpha to efficiently generate disease. PMID- 16473274 TI - How to decrease p27Kip1 levels during tumor development. AB - p27Kip1, a cyclin-cdk inhibitor, is a tumor suppressor. An overwhelming amount of data correlate p27 abundance to tumor prognosis in humans. Mouse models have supported the importance of decreasing p27 to tumor incidence. Inactivation of most tumor suppressors occurs at the level of gene mutation or silencing, but p27 is regulated posttranscriptionally, and how its level is reduced in cancer is largely unknown. Reports on a series of allelic mice with p27 mutations affecting different posttranscriptional regulatory pathways are emerging and being used to examine which pathways are necessary for p27 turnover associated with tumor development, with surprising results. PMID- 16473275 TI - TORgeting oncogene addiction for cancer therapy. AB - The PI3K-Akt-mTOR growth-regulating pathway is conserved from mammals to flies and hyperactivated in many cancers. Accordingly, rapamycin analogs, which are inhibitors of mTOR-Raptor signaling, have recently garnered much interest as potential therapeutic agents against cancer. However, due to the heterogeneity of tumors, prior knowledge of the genetic and biochemical background of cancer cells will be required for effective targeted therapy. Thus, the identification of biological markers against activated oncogenic pathways is needed. In the January issue of Nature Medicine, Thomas et al. identify the loss of VHL tumor suppressor gene as a potential determining factor in tumor sensitivity to rapamycin. PMID- 16473276 TI - Forced retinoic acid receptor alpha homodimers prime mice for APL-like leukemia. AB - RARA becomes an acute promyelocytic leukemia (APL) oncogene by fusion with any of five translocation partners. Unlike RARalpha, the fusion proteins homodimerize, which may be central to oncogenic activation. This model was tested by replacing PML with dimerization domains from p50NFkappaB (p50-RARalpha) or the rapamycin sensitive dimerizing peptide of FKBP12 (F3-RARalpha). The X-RARalpha fusions recapitulated in vitro activities of PML-RARalpha. For F3-RARalpha, these properties were rapamycin sensitive. Although in vivo the artificial fusions alone are poor initiators of leukemia, p50-RARalpha readily cooperates with an activated mutant CDw131 to induce APL-like disease. These results demonstrate that the dimerization interface of RARalpha fusion partners is a critical element in APL pathogenesis while pointing to other features of PML for enhancing penetrance and progression. PMID- 16473277 TI - Forced homo-oligomerization of RARalpha leads to transformation of primary hematopoietic cells. AB - Almost 100% of APL patients carry chimeric transcripts encoding truncated RARalpha fused to homo-oligomerization domains from partner proteins. To gain further insights into the cellular transformation mechanisms mediated by RARalpha fusion proteins, thorough structure/function analyses have been performed and identified the POZ homo-oligomerization domain as the minimal transformation domain that is necessary and sufficient for PLZF-RARalpha-mediated in vitro transformation of primary hematopoietic cells. A transformation-incompetent PLZF RARalpha mutant defective in homo-oligomerization but not corepressor interaction could be rescued by synthetic FKBP-oligomerization domains. Furthermore, an artificial FKBP-RARalpha construct not only mimicked various biochemical properties of bona fide RARalpha fusion proteins but also mediated an ATRA dependent transformation. Taken together, these findings endorse an oligomerization-dependent mechanism for RARalpha-mediated transformation and suggest a potential avenue for molecular therapy. PMID- 16473278 TI - Block of T cell development in P53-deficient mice accelerates development of lymphomas with characteristic RAG-dependent cytogenetic alterations. AB - Mice deficient in the DNA damage sensor P53 display normal T cell development but eventually succumb to thymic lymphomas. Here, we show that inactivation of the TCR beta gene enhancer (E beta) results in a block of T cell development at stages where recombination-activating genes (RAG) are expressed. Introduction of the E beta mutation into p53-/- mice dramatically accelerates the onset of lethal thymic lymphomas that harbor RAG-dependent aberrant rearrangements, chromosome 14 and 12 translocations, and amplification of the chromosomal region 9A1-A5.3. Phenotypic and genetic analyses suggest that lymphomas emerge through a normal thymocyte development pathway. These findings provide genetic evidence that block of lymphocyte development at stages with RAG endonuclease activity can provoke lymphomagenesis on a background with deficient DNA damage responses. PMID- 16473279 TI - X chromosomal abnormalities in basal-like human breast cancer. AB - Sporadic basal-like cancers (BLC) are a distinct class of human breast cancers that are phenotypically similar to BRCA1-associated cancers. Like BRCA1-deficient tumors, most BLC lack markers of a normal inactive X chromosome (Xi). Duplication of the active X chromosome and loss of Xi characterized almost half of BLC cases tested. Others contained biparental but nonheterochromatinized X chromosomes or gains of X chromosomal DNA. These abnormalities did not lead to a global increase in X chromosome transcription but were associated with overexpression of a small subset of X chromosomal genes. Other, equally aneuploid, but non-BLC rarely displayed these X chromosome abnormalities. These results suggest that X chromosome abnormalities contribute to the pathogenesis of BLC, both inherited and sporadic. PMID- 16473280 TI - Using small molecules to overcome drug resistance induced by a viral oncogene. AB - We used small molecule screening to discover compounds and mechanisms for overcoming E6 oncogene-mediated drug resistance. Using high-throughput screening in isogenic cell lines, we identified compounds that potentiate doxorubicin's lethality in E6-expressing colon cancer cells. Such compounds included quaternary ammonium salts, protein synthesis inhibitors, 11-deoxyprostaglandins, and two additional classes of compounds-analogs of 1,3-bis(4-morpholinylmethyl)-2 imidazolidinethione (a thiourea) and acylated secondary amines that we named indoxins. Indoxins upregulated topoisomerase IIalpha, the target of doxorubicin, thereby increasing doxorubicin lethality. We developed a photolabeling strategy to identify targets of indoxin and discovered a nuclear actin-related protein complex as a candidate indoxin target. PMID- 16473283 TI - The unique clinical presentation of children with chronic arthritis: putting the pediatrics in pediatric rheumatology. PMID- 16473284 TI - Genetic analysis of juvenile rheumatoid arthritis: approaches to complex traits. PMID- 16473285 TI - Gene expression arrays in juvenile rheumatoid arthritis: will the blind men finally see the elephant? PMID- 16473286 TI - Emerging therapies in juvenile rheumatoid/idiopathic arthritis. PMID- 16473287 TI - The future of rheumatology research: the Childhood Arthritis and Rheumatology Research Alliance. PMID- 16473291 TI - The biopsychosocial approach to adolescents with somatoform disorders. AB - Somatoform disorders are presented in the first article in this issue of Adolescent Medicine Clinics because the physical symptoms that cause the adolescent to present for diagnosis and treatment reflect the interaction of the psyche and the soma in ways that are poorly understood. Because of dualistic conceptualizations that are encouraged by technology such as MRI, CT scans and other technologically advanced tools, patients who have these conditions often suffer. As noted by Cassell [35], "suffering is experienced by persons, not merely by bodies, and has its source in challenges that threaten the intactness of the person as a complex social and psychological entity." Clinicians who care for adolescents who have somatic complaints also suffer when they are unable to provide relief of an adolescent's suffering. Cassell [36] noted that "physicians are less skilled at what were once thought to be the basic skills of doctors discovering the history of an illness though questioning and physical examination, and working toward healing the whole person." The biopsychosocial approach offers a means of working toward healing the whole person, and the focus of this article is on practical solutions to difficult challenges that are presented by adolescents and their families. PMID- 16473292 TI - Consent, competence, and confidentiality related to psychiatric conditions in adolescent medicine practice. AB - Health care for adolescents with psychiatric conditions plays out on a complex stage with considerable state variation, based on a mix of statutory and case law. Added to this are less defined factors such as level of trust in community providers, level of cooperation between generalists and specialists, and local regulatory stances toward adolescent health care and mental health care. And, of course, there is the great diversity in diagnosis and maturity level, as well as family cohesion, from patient to patient (and even within a given patient across time). Finally, this situation resides within a larger environment of stigma vis a-vis mental health care, most notably evident in the United States in disparate insurance coverage of mental versus physical health treatment. With so complex and varied a picture, clinicians should consult with legal counsel to understand applicable state law and local regulatory guidance (if any)and should also seek out ethical consultation when law does not apply or is not decisive, leaving ongoing concerns. And, as with clinical decision-making, in law and ethics "facts matter" [4]. A growing body of law carves out exceptions to general requirements for parental consent, including in mental health care. Ethically informed discussion around capacity determinations, the consent process, and confidentiality can help engage adolescents as "emerging adults"while remaining mindful of risky behavior and "immediate future" orientation that can be hallmarks of adolescence [30]. Respect for the adolescent, parental responsibility toward their child's best interests, and the family unit generally are paramount. Respect--coupled with caution, greater disclosure and cultural sensitivity, and a participatory approach to decision-making that seeks out the least restrictive and coercive options-can help avoid potential legal traps. How best to proceed? It truly depends-with law and ethics the start (not end) of the discussion and analysis. PMID- 16473293 TI - Medical conditions with psychiatric manifestations. AB - A variety of medical conditions can present, or be associated, with psychiatric symptoms. At times, these may be so prominent that they can overshadow the underlying pathophysiologic process that accounts for them. Thus, it is equally important for mental health providers to be alert to the possibility that adolescents whom they are treating may have symptoms related to a treatable medical condition as it is for primary care providers to conduct a targeted history and physical examination with their adolescent patients exhibiting psychiatric symptoms. Using the biopsychosocial approach, these two domains are not considered separately or hierarchically, but as highly interactive. In some cases (eg, adrenal insufficiency), appropriate and continued treatment of the underlying condition results in resolution of the psychiatric symptoms. In others (eg, SLE), treatment of the underlying condition may alleviate but may also exacerbate psychiatric symptoms. Therefore, comprehensive treatment of adolescents with psychiatric symptoms due to a medical condition may require the professional services of primary care, mental health, and specialty care providers, but their services should follow the collaborative model espoused throughout the other articles in this volume. PMID- 16473294 TI - Disorders of mood and anxiety in adolescents. AB - Mood disorders and anxiety disorders contribute significantly to morbidity and mortality during adolescence. These disorders often persist or recur in adulthood. Clinical presentations in the primary care setting are myriad and often confusing. Early recognition, differentiation from physical and other psychiatric disorders, and accurate diagnosis lead to more appropriate treatment and improved outcome. Collaboration among primary care providers and mental health professionals is key to reducing the suffering from these disorders for adolescents and their families. PMID- 16473295 TI - Disruptive behaviors: conduct and oppositional disorders in adolescents. AB - CD and ODD present unique challenges in both the diagnostic and treatment arenas. The complex network of causes leading to disruptive disorders makes it necessary to structure treatment in a multifaceted manner. The earlier that adolescents are identified as exhibiting disruptive behavior disorders, the better the opportunity to intervene. While early-onset CD is very difficult to treat effectively, the disorder is far more difficult to correct if left untreated and can ultimately lead to ASPD in adulthood. When physicians are presented with behavioral complaints, they must fully assess the adolescent to determine what, if any, referrals and treatments are needed. PMID- 16473296 TI - Adolescent personality disorders in adolescent medicine. AB - Both BPD and ASPD are disorders whose complex presentations pose numerous clinical challenges to all health care professionals. However, early careful diagnosis and management can diminish the long-term morbidity of these illnesses and offer the hope of a better outcome. As data emerge regarding the long-term prognosis and effective treatment of PDs, clinicians are cautioned to avoid the traditional teaching that the dysfunctional patterns will necessarily continue into adulthood. PMID- 16473297 TI - Psychosis in adolescence. AB - Psychotic behavior in adolescent patients is uncommon but occasionally encountered in adolescent medicine practice. Although a physician may not be able to distinguish the precise diagnosis initially, immediate symptomatic management of acute psychotic symptoms is indicated to allay the patient's anxiety and tQ begin treatment. In chronic and more subtle conditions, their recognition is the essential first step. In all cases, referral to a mental health specialist is indicated for comprehensive evaluation and appropriate management. Continuing support of patient and family by the primary care physician avoids a perception of abandonment and can help ensure effective long-term care. PMID- 16473298 TI - Psychological testing in adolescent medicine. AB - Psychological and neuropsychological assessment remains an important aspect of clinical evaluation in adolescents with psychiatric and neurologic disorders. The primary care practitioner can refer for psychological and neuropsychological assessment when cognitive, behavioral, or psychiatric problems appear to be affecting the adolescent's learning, psychosocial development, or overall functioning. The most appropriate assessment depends upon the diagnostic issue at hand. Most referrals will include IQ and achievement testing to assess for learning disabilities; behavioral and personality assessment to aid in psychiatric diagnosis and treatment planning; and neuropsychological testing for more complex issues, such as the impact of neurological disorders and injuries on cognition and behavior. The patient can be referred to the school psychologist for IQ and achievement testing that complements any additional testing that may be required to help determine the best intervention. Psychological assessment can be invaluable in the early identification and intervention of learning, behavioral, and psychiatric difficulties in adolescents. PMID- 16473299 TI - Psychopharmacology in adolescent medicine. AB - Psychopharmacology is a challenge for health care providers treating adolescents. A detailed and accurate assessment, including developmental issues relevant to adolescence in general and to the individual adolescent, guides clinicians in formulating thoughtful and effective treatment plans to meet the needs of each patient. Parents play an important role in providing family history regarding psychiatric diagnoses and the response to various drugs, in making decisions to initiate medication and to change a medication regimen, and in monitoring an adolescent's adherence to a prescribed regimen. The role of parents is especially important for younger patients. Following the biopsychosocial model, rarely should psychopharmacologic agents be used as the sole means to treat a psychiatric condition in adolescents. Pharmacologic agents described in this article are tools that have their effect in the biological domain of central neurotransmitters, but psychosocial interventions addressing the emotional and behavioral issues that are the indications for such medication are generally also required. The development of newer medications holds promise for more effective treatment of target symptoms with minimal side effects. PMID- 16473300 TI - Psychiatric emergencies in adolescents. AB - The spectrum of psychiatric emergencies in adolescents may be best appreciated using the broad framework of urgency. ED physicians and staff using such a framework will be in a better position to triage and to tailor the evaluation assessment and target the intervention and disposition. Understanding the range of urgency can minimize frustration, enhance the clinician's ability to accurately assess complex situations, and make a tremendous difference in the patient's receipt of future services. This article reviews the epidemiology, risk factors, and critical elements of emergency evaluation and treatment of a variety of juvenile psychiatric emergencies. Factors influencing presentation, disposition,and consideration of local treatment resources are reviewed. PMID- 16473301 TI - Adolescents' use of the Internet: a controversial, coming-of-age resource. AB - Emerging research on adolescents and the Internet should be thoughtfully considered, especially with respect to the influence of Internet activities on mental health and psychiatric conditions. A given Internet site may have content that would be helpful to one, have no effect on another, and be harmful to yet an other adolescent. The effect on the adolescent would in part be determined by the site itself and the way in which it provides information and interactivity, and in part on the characteristics of the particular adolescent. It will be necessary to employ diverse approaches to keep current the information on how adolescents use the Internet, as well as to understand how it affects them. Rather than rely on focus groups and self-report surveys, researchers should examine adolescent Internet use with naturalistic methods that will provide quantitative and qualitative data regarding this new domain. Additionally, researchers can use experimental designs to explore how Internet experiences or exposure to online information affects health knowledge, attitudes, and behaviors. Because adolescents rely on the Internet for both its content and technology,youth caregivers and advocates should continue to research and study this medium. The efficacy of delivering Internet-based therapies and prevention pro-grams should be studied. Mental health organizations should remain vigilant,monitoring and helping to improve websites [7]. Parents should also be aware of their adolescents' online activities, so it may be necessary for health practitioners to educate parents about what is available and occurring on line. Lastly, youth need to be better informed on how to best use the Internet. We take for granted that adolescents are savvy users of this technology, when it is not yet fully known how young people make use of the Internet and how they incorporate information from the Internet into their lives. Online media literacy skills are necessary and should be developed among the very young to make the best use of the Internet. PMID- 16473302 TI - Individual, family, and group therapy for adolescents. AB - The three main psychotherapeutic treatment modalities include individual,family, and group therapies. Many theoretic orientations guide psychotherapists as they try to help adolescents with mental health problems. PCPs play a critical role in initial assessment of mental health symptoms, in addition to coordinating treatment needs. There is a need for increased education regarding mental health treatment for health care providers to help them connect adolescents and their families to appropriate mental health care providers. Integrative approaches that involve more than one treatment modality are often needed to provide the best treatment for adolescents. Better collaborative care not only improves physician understanding of mental health treatment but also improves the mental health provider's understanding of the medical system [30]. This emerging con-text of increased mutual collaborative care builds a better system that serves the adolescent.Web-based resources related to psychotherapy for adolescents American Academy of Child and Adolescent Psychiatry http://www.AACAP.org American Association for Marriage & Family Therapy http://www.AAMFT.org American Psychological Association http://www.APA.org American Psychiatric Association http://www.psych.org National Mental Health Association http://www.NMHA.org National Alliance for the Mentally Ill http://www.NAMI.org PMID- 16473303 TI - Treatment settings for adolescent psychiatric conditions. AB - The intensive psychiatric treatment settings, including inpatient hospitalization, day programs and residential care, are valuable options for clinicians caring for adolescents with serious mental health problems. The availability of these setting may be limited by geographic, insurance, or financial restraints, but providers should stay informed about the treatment settings available in their area and be prepared to advocate for their adolescent patients' psychiatric needs. Although little evidence-based practice is available to guide clinicians taking care of adolescents in need of the most intensive psychiatric treatment settings, certain elements of care have proven most essential, including especially the successful engagement of families in treatment. Good outcomes for the most complicated adolescents follow from successful collaborations with families and the various providers that intersect in their multidisciplinary care. PMID- 16473304 TI - The Fragile X premutation: new insights and clinical consequences. AB - The Fragile X syndrome (FXS, MIM 309550) is still the most prevalent cause of heritable mental retardation, with a frequency of 1/4000 males and 1/6000 females. The syndrome and its particular pattern of heredity are caused by a dynamic mutation, involving an unstable expansion of a trinucleotide (CGG) repeat at the 5' UTR of the FMR1 gene, located at Xq27.3. Expansion of this repeat region greater than 200 repeats leads to methylation-coupled silencing of the gene and absence of the Fragile X mental retardation protein (FMRP), causing the classical FXS. Individuals with expanded repeat lengths varying from 50 to 200 repeats do not exhibit the classical FXS phenotype, but are considered as fragile X premutation (PM) carriers. These premutation alleles may become unstable, only through maternal transmission, with further expansion in the next generations. For long-time, male and female premutation carriers were considered as asymptomatic. This view was, however, gradually challenged with the description and reports of different premutation-associated clinical phenotypes over the last decade. PMID- 16473305 TI - Spectrum and distribution of MECP2 mutations in 424 Rett syndrome patients: a molecular update. AB - Mutations in the MECP2 (Methyl-CpG-binding protein) gene have been reported to cause Rett syndrome (RTT), an X-linked progressive encephalopathy. Recent studies have identified large gene rearrangements that escape the common PCR-based mutation screening strategy and mutations in a novel MeCP2 isoform (named MECP2B). We have collected the results of MECP2 mutational analysis concerning 424 RTT patients conducted in eight laboratories in France. In total, 121 different MECP2 mutations were identified. R168X (11.5%) is the most common of MECP2 mutations, followed by R270X (9%), R255X (8.7%), T158 M (8.3%) and R306C (6.8%). Only eight mutations had relative frequency>3%. Large and complex rearrangements not previously detected using only a PCR-based strategy represent 5.8% of MECP2 mutations. On the contrary, mutation in exon 1 appears to be rare (less than 0.5%). These data demonstrate the high allelic heterogeneity of RTT in France and suggest that routine mutation screening in MECP2 should include quantitative analysis of the MECP2 gene. This study represents an important instrument for molecular diagnosis strategy and genetic counseling in RTT families. PMID- 16473306 TI - Virilization of the external genitalia and severe mental retardation in a girl with an unbalanced translocation 1;18. AB - A female infant with dysmorphic facial features, psychomotor retardation, and clitoris hypertrophy is described. Molecular cytogenetic analyses revealed a de novo unbalanced translocation, causing partial monosomy 1p36 and partial trisomy 18q22. Monosomy 1p was confirmed by FISH, and trisomy of the distal part of chromosome 18q was demonstrated by microFISH. Gene copy number changes in these chromosomal regions were determined by array-CGH. The absence of a number of facial dysmorphic signs, and the presence of clitoris hypertrophy indicate that the combination of a del(1p36->pter) with a dup(18q22->qter) may lead to a unique phenotypic constellation. The findings at birth and at age 12 years in our patient are compared with genotype-phenotype correlations discussed in the literature. PMID- 16473307 TI - Identification and characterization of a novel variant in the highly conserved catalytic center of Rab11a. AB - Small GTPases of the Rab family regulate vesicular traffic and distribution of proteins in different cell types. Rab11a is a member of this GTP hydrolyzing protein class and acts as a mediator of insulin stimulated translocation of the glucose transporter GLUT4 in peripheral tissues including heart and skeletal muscle. Here we report on Rab11a Q70R, a mutation in the catalytic center of Rab11a, observed in the cardiomyoblast cell line H9c2. Analysis of GTPase activity showed that Rab11a Q70L acts as a classical constitutive active mutant. Interestingly, the GTPase activity of Rab11a Q70R was not significantly different from the enzymatic activity of the Rab11a Q70 wild type protein. We therefore conclude that the glutamine residue of Rab11a at position 70 is not strictly essential for GTPase activity of this protein in contrast to Ras and other Rab proteins. PMID- 16473309 TI - HLA genes in Southern Tunisians (Ghannouch area) and their relationship with other Mediterraneans. AB - South Tunisian HLA gene profile has studied for the first time. HLA-A, -B, -DRB1 and -DQB1 allele frequencies of Ghannouch have been compared with those of neighboring populations, other Mediterraneans and Sub-Saharans. Their relatedness has been tested by genetic distances, Neighbor-Joining dendrograms and correspondence analyses. Our HLA data show that both southern from Ghannouch and northern Tunisians are of a Berber substratum in spite of the successive incursions (particularly, the 7th-8th century A.D. Arab invasion) occurred in Tunisia. It is also the case of other North Africans and Iberians. This present study confirms the relatedness of Greeks to Sub-Saharan populations. This suggests that there was an admixture between the Greeks and Sub-Saharans probably during Pharaonic period or after natural catastrophes (dryness) occurred in Sahara. PMID- 16473308 TI - HLA non-class II genes may confer type I diabetes susceptibility in a Mapuche (Amerindian) affected family. AB - A rare case of type I diabetes is studied in an Amerindian (Mapuche) family from Chile, analyzing glutamic acid decarboxylase, islet-cell autoantibodies and human leukocyte antigen (HLA) genes. The affected sib is the only one that has one specific HLA haplotype combination that differs from the other sibs only in the HLA class I genes. It is concluded that HLA diabetes susceptibility factors may be placed outside the class II region or even that susceptibility factors do not exist in the HLA region in this Amerindian family. PMID- 16473310 TI - No mutations detected in the INSR gene in a chromosome 19p13 linked migraine pedigree. AB - The aim of this study was to investigate through direct sequencing the insulin receptor (INSR) gene in DNA samples from a migraine affected family previously showing linkage to chromosome 19p13 in an attempt to detect disease associated mutations. Migraine is a common debilitating disorder with a significant genetic component. At present, the number and type of genes involved in the common forms of migraine are not clear. The INSR gene on chromosome 19p13.3-13.2 is a gene of interest since a number of single nucleotide polymorphisms (SNPs) located within the gene have been implicated in migraine with (MA) and without aura (MO). Six DNA samples obtained from non-founding migraine affected members of migraine family 1 (MF1) were used in this study. Genomic DNA was sequenced for the INSR gene in exons 1-22 and the promoter region. In the six migraine family member samples, previously reported SNPs were detected within two exonic DNA coding regions of the INSR gene. These SNPs, in exons 13 and 17, do not alter the normal INSR polypeptide sequence. In addition, intron 7 also revealed a DNA base sequence variation. For the 5' untranslated promoter region of the gene, no mutations or polymorphisms were detected. In conclusion, this study detected no INSR mutations in affected members of a chromosome 19 linked migraine pedigree. Hence, migraine linkage to this chromosomal region may involve other candidate genes. PMID- 16473311 TI - A different approach to telomere analysis with ddPRINS in chronic lymphocytic leukemia. AB - Telomeric sequences, located at the very end of the chromosomes, compensate for the chromosomal shortening as it happens after each round of cell division. Telomeric sequences influence the progress of cellular senescence and cancer progression. It has been reported that telomeres are shortened in acute leukemias where the cell turnover is high. B-cell chronic lymphocytic leukemia (CLL) is a particularly interesting haematological malignancy in regard to telomere dynamics because most of the malignant cells in CLL are mitotically inactive. In this study, we analysed the telomere length in patients with B-cell CLL in a comparison with the control group by using ddPRINS technique. Twenty patients with CLL and four healthy donors as a control group were included. We found short telomeres and no detectable telomeric repeats at the sites of chromosome fusion. We hypothesise that the telomeric erosion in CLL may reflect the dominance of malignant cells with an abnormally long life span. These cells may have encountered many antigenic stimulants in the past and hence underwent multiple clonal expansions. Our findings imply that shortened telomeres in CLL may be reflecting the "history" of the disease and serve as an independent prognostic factor. PMID- 16473312 TI - Another observation with VATER association and a complex IV respiratory chain deficiency. AB - The VATER association of vertebral anomalies (V), anal atresia (A), esophageal atresia and/or tracheo-esophageal fistula (TE), radial and renal anomalies (R) is a common congenital association of unknown origin with probably heterogeneous causes. Here, we report on a girl presenting with pre- and postnatal growth retardation, esophageal atresia, vertebral and costal anomalies and a unilateral radial defect, consistent with the diagnosis of VATER association. In the first month of life, she presented with failure to thrive, severe episodes of hypoglycemia, liver dysfunction and high levels of lactate, which prompted us to perform metabolic screening. A complex IV respiratory chain deficiency (RCD) was diagnosed on a liver biopsy. The respiratory chain defect was not observed in skin fibroblasts. No mtDNA point mutation or deletion was identified. The girl is now 9 years old and has a normal mental development but persistent feeding difficulties and moderate hyperlactatemia. To our knowledge, this is the second report of VATER association with mitochondrial disorder. In a previous report, a VACTERL association was observed in a girl with the mitochondrial A3243G point mutation. The association of VATER phenotype with a mitochondrial disorder may be coincidental but could also suggest that the presence of multiple malformations is the result of the antenatal expression of RCD. PMID- 16473313 TI - Central precocious puberty in a girl with Williams syndrome: the result of treatment with GnRH analogue. AB - Williams syndrome (WS) is a well-known microdeletion syndrome characterized by specific facial features, retardation in growth and development, typical personality and cardiac defects. Poor growth potential is further affected by central precocious puberty (CPP) which is frequent in these patients. A WS patient with CPP is presented, whose pubertal development and bone age progression were arrested by administration of GnRH analogues. The case is reported to discuss the role of GnRH analogues for management of CPP in patients with WS. PMID- 16473315 TI - Cri du chat syndrome determined by the 5p15.3-->pter deletion--diagnostic problems. AB - A cytogenetic analysis was performed on an 8-day-old girl, who was suspected of Cri du chat syndrome (CdCS) on the basis of a cat-like cry, despite her dysmorphic features not being characteristic of this syndrome. The cytogenetic analysis revealed a partial deletion of the short arm of chromosome 5, but did not allow precise specification of the break points. Fluorescence in situ hybridization (FISH) analysis, using the specific probe for CdCS, revealed two signals in all the cells analyzed. However, one of two signals was less intense than the other. Thus, telomere probes were applied for all chromosomes. Two signals from 5q and one signal from 5p were observed. The results allowed us to establish the location of the deleted fragment as 5p15.3-->5pter [46,XX,del(5)(p15.3)]. The analysis of a genotype-phenotype correlation confirmed that the cat-like cry, but not the characteristic dysmorphic features of CdCS are correlated with the deletion of 5p15.3. PMID- 16473314 TI - Fibular aplasia with ectrodactyly--broadening the clinical spectrum. AB - Fibular aplasia-ectrodactyly is a rare disorder of the central axis, characterized by shortening of the affected limbs and formation of split hand and/or foot. Here we report on a severely affected case of fibular aplasia with ectrodactyly, in which the upper limb malformations are more pronounced than usually described in sporadic cases. PMID- 16473316 TI - A further case of opsismodysplasia with hydrocephalus. AB - We present a case of opsismodysplasia, a very rare skeletal dysplasia, in a term newborn female who had short length, short extremities and markedly short fingers. Radiological studies demonstrated severe platyspondyly, absence of epiphyseal ossification centers, short tubular bones, especially severe in hands and feet, with metaphyseal cupping. She also had hydrocephaly, a rare finding in opsismodysplasia. In our literature review we have found 24 cases, 17 born alive and seven terminations of pregnancy (TOPs). PMID- 16473318 TI - Hepatic stellate cells in human schistosomiasis mansoni: a comparative immunohistochemical study with liver cirrhosis. AB - This study compares the populations of liver mesenchymal cells (LMCs) and their proliferative activity in schistosomal periportal fibrosis and in hepatitis C virus-induced cirrhosis. LMCs were evaluated by immunohistochemical double staining for proliferating cell nuclear antigen (PCNA) and alpha-smooth muscle actin (alpha-SMA) or glial fibrillary acid protein (GFAP) in liver biopsies from humans with schistosomal fibrosis (n=40), hepatitis C virus-induced cirrhosis (n=20), and normal controls (n=20). The number of LMCs was found to be higher in schistosomal fibrosis than in the normal liver, but lower than in cirrhosis. alpha-SMA- and GFAP-positive cells were increased in both diseases, but more so in cirrhosis. In cirrhotic liver, alpha-SMA-positive cells were highly predominant in relation to GFAP-positive cells. However, there was an inverted ratio between these cells in schistosomiasis as compared to cirrhosis. The PCNA labeling index was higher in alpha-SMA-positive cells than in GFAP-positive cells, and did not differ between pipe-stem fibrosis and liver cirrhosis regarding alpha-SMA- or GFAP-positive cells. The predominance of GFAP-positive cells observed in schistosomiasis suggests that hepatic stellate cells (HSCs) have a major role in connective tissue deposition in the human schistosomal liver. On the other hand, the smaller number of LMCs in schistosomal fibrosis in comparison to liver cirrhosis may be related to mild and limited injury due to the schistosomal egg-induced inflammatory response. The granulomatous inflammation around Schistosoma mansoni eggs appears to mobilize and activate a reduced number of mesenchymal cells in comparison to the scattered necro inflammatory reaction produced by the hepatitis C virus. PMID- 16473319 TI - A quantitative high-throughput trapping assay as a measurement of potential for bioactivation. AB - Idiosyncratic adverse drug reactions (ADRs) are one of the most common causes of pharmaceutical withdrawals and labeling changes. Most ADRs are caused by drugs that form reactive species that can bind covalently to macromolecules such as proteins. The current methodology for the measurement of covalent binding relies on the use of radiolabeled material that requires an investment in time and resources not typically expended until later in the discovery process. Efforts are also made to identify reactive intermediates by the use of chemical trapping agents, such as reduced glutathione and cyanide, to form stable adducts that are characterized by liquid chromatography-tandem mass spectrometry and/or nuclear magnetic resonance spectroscopy. Here, we describe a high-throughput assay for the measurement of reactive intermediate formation. The method involves incubation of cold compound with liver microsomes in the presence of [14C]potassium cyanide. Hard electrophilic species would react with the trapping agent, resulting in the formation of a radiolabeled conjugate. Unreacted trapping agent is removed using solid-phase extraction, and the amount of radiolabeled conjugate present is determined by liquid scintillation counting. This newly developed screen has proved to be specific, sensitive, robust, and a powerful tool for assessing bioactivation potential. PMID- 16473320 TI - Trizol-based method for sample preparation and isoelectric focusing of halophilic proteins. AB - A persisting complication in the development of well-resolved two-dimensional PAGE maps of halophilic proteins is their natural incompatibility with isoelectric focusing (IEF). The complete desalting of samples, which is necessary for IEF, tends to aggregate halophilic proteins, often requires relatively large amounts of starting material due to significant loss of sample, and is relatively time-consuming. Here, we describe a method of preparing protein samples from the haloarchaeon Haloferax volcanii that not only desalts the samples thoroughly but also drastically reduces the amount of protein loss associated with previous sample preparation methods and prevents protein aggregation during the removal of salt. This method of sample preparation, which incorporates Trizol (phenol/guanidine isothiocyanate), can easily be extended to analyze halophilic proteins from other organisms. PMID- 16473321 TI - Development and initial assessment of objective fatigue measures for apple harvest work. AB - Previous research has shown that neck, back and shoulder musculoskeletal strain is a major occupational health problem affecting migrant orchard harvest workers. Researchers seek to measure the effect of an ergonomic modification to the apple picking bucket on muscle fatigue, however objective measures for use in the orchard are not yet available. The purpose of this study is to develop simple back, shoulder or arm strength measures, which detect statistically significant drops in strength over one workday. Candidate muscle strength measures were piloted in the laboratory, adapted for the orchard and evaluated (n=102). Data were analyzed for morning to afternoon fatigue, and for correlation between fatigue score and hours worked. In the laboratory, the timed arm hold (35.7% time reduction, 95% CI: 21.81-49.61), and the timed spinal extension (31.8% time reduction, 95% CI: 23.54-39.96) showed significant fatigue. In the orchard (n=102), only the timed arm hold showed significant (11.4%, p<.0001) fatigue. The potential effect of field conditions and subject motivation on these results needs further exploration. PMID- 16473322 TI - Mitochondrial DNA maintenance and bioenergetics. AB - Oxidative phosphorylation requires assembly of the protein products of both mitochondrial and of nuclear genomes into functional respiratory complexes. Cellular respiration can be compromised when mitochondrial DNA (mtDNA) sequences are corrupted. Oxidative damage resulting from reactive oxygen species (ROS) produced during respiration is probably a major source of mitochondrial genomic instability leading to respiratory dysfunction. Here, we review mechanisms of mitochondrial ROS production, mtDNA damage and its relationship to mitochondrial dysfunction. We focus particular attention on the roles of mtDNA repair enzymes and processes by which the integrity of the mitochondrial genome is maintained and dysfunction prevented. PMID- 16473323 TI - Effects of trehalose on the phase behavior of DPPC-cholesterol unilamellar vesicles. AB - A systematic study is presented of the effects of trehalose on the physical properties of extruded DPPC-cholesterol unilamellar vesicles. Particular emphasis is placed on examining how the interactions present in the hydrated state translate into those in the dehydrated state. Observations from HSDSC and DSC are used to examine the phase behavior of hydrated and dehydrated vesicles, respectively. The concentration of trehalose inside and outside the vesicles is manipulated, and is shown to affect the relative stability of the membranes. Our results show for the first time that a combination of high inner and low outer trehalose concentration is able to decrease the gel-to-liquid crystalline phase temperature (T(m)), while any other combination will not. Upon dehydration, the T(m) of all lipid mixtures increases. The extent of the increase depends on the trehalose distribution across the bilayer. The T(m) changes in the same direction with trehalose concentration for both freeze-dried and fully hydrated samples, suggesting that the trehalose distribution across the vesicle membrane, as well as the trehalose-phospholipid interaction, is maintained upon lyophilization. The results presented in this work may aid in the formulation of systems to be used in the lyophilization of liposomes for drug delivery applications. PMID- 16473324 TI - Spectroscopic studies of amphotericin B solubilized in nanoscale bilayer membranes. AB - Nanodisks (ND) are discrete nanometer scale phospholipid bilayers whose perimeter is circumscribed by amphipathic apolipoproteins. The membranous environment of ND serves as a matrix for solubilizing the polyene antibiotic amphotericin B (AMB). The spectral properties of AMB in ND are dependent upon AMB concentration. Whereas AMB-ND prepared at a concentration of 2.5 mg AMB per 10 mg phospholipid are consistent with AMB self association in the ND membrane environment, AMB-ND prepared at 0.25 or 0.025 mg AMB per 10 mg phospholipid give rise to spectra reminiscent of AMB in organic solvent. Incubation of ND prepared at a phospholipid/AMB ratio of 400:1 (w/w) at 37 degrees C for 1 h induced a shift in absorbance and near UV circular dichroism spectra consistent with antibiotic self association. The kinetics of this spectral transition were investigated as a function of incubation temperature. While no change in A388 nm occurred in incubations at 20 degrees C, a time-dependent decrease in A388 nm was observed at 25, 30 and 37 degrees C. Inclusion of ergosterol in the ND membrane attenuated temperature-induced AMB spectral changes. In Saccharomyces cerevisiae growth inhibition assays, ND containing self associated AMB were somewhat less effective than ND possessing a greater proportion of monomeric AMB. On the other hand, inclusion of ergosterol or cholesterol in the ND particle did not alter the growth inhibition properties of AMB-ND. The miniature membrane environment of ND provides a novel milieu for solubilization and characterization of lipophilic biomolecules. PMID- 16473325 TI - The role of imagery in the maintenance and treatment of snake fear. AB - Two studies assessed the role of mental imagery in the maintenance of fear of common phobic stimuli. Study 1 asked participants who were afraid of a wide range of phobic stimuli to report their visual and somatic imagery. Blind ratings of the imagery on horror and vividness were positively correlated with participant's self-reported fear and avoidance. Study 2 tested the efficacy of modifying imagery using cognitive restructuring compared to in vivo exposure and a minimal exposure, relaxation control in snake fearful participants. Both active treatment groups improved significantly more than the control group in self-reported snake fearfulness and behavioral approach. Condition also interacted with initial severity. Highly fearful subjects responded better to the cognitive imagery modification than to the in vivo exposure, and found the cognitive intervention significantly less aversive. These results are seen as supporting a cognitive model of the maintenance of specific phobia. PMID- 16473326 TI - Sensory-evoked potentials recordings from the ventral tegmental area, nucleus accumbens, prefrontal cortex, and caudate nucleus and locomotor activity are modulated in dose-response characteristics by methylphenidate. AB - Most of the studies investigating the effects of methylphenidate (MPD) are using behavioral and biochemical approaches. There are some electrophysiological studies about the effects of MPD on spontaneous electrical activity; however, there is none about the effects of MPD on sensory inputs. The objectives of the present study were to investigate the MPD dose-response characteristics on locomotor activity and sensory inputs using acoustic stimuli. Freely behaving rats previously implanted with semi-microelectrodes were used to record from four brain areas known to be sites of psychostimulant action. For locomotor behavior assessment, rats received saline on experimental day 1 and an acute administration of MPD (0.6, 2.5, or 10.0 mg/kg, i.p.) on experimental day 2. Using an automated, computerized activity-monitoring system, locomotor behavior was recorded for 2-h postinjection on both days. For the electrophysiological experiments, animals were implanted with permanent electrodes in the ventral tegmental area (VTA), nucleus accumbens (NAc), prefrontal cortex (PFC), and caudate nucleus (CN) under general anesthesia. Five to seven days after electrode implantation, they were used to study the effects of three different MPD doses on the response to sensory inputs. The lowest dose of MPD (0.6 mg/kg, i.p.) failed to alter locomotor activity, while the two higher MPD doses (2.5 and 10.0 mg/kg) elicited increase in locomotion, with the 10.0 mg/kg dose increased at least twice as much as the 2.5 mg/kg dose. However, the same three MPD doses elicited significant attenuation of sensory inputs in dose-response characteristics, i.e., as MPD dose increased, evoked sensory inputs decreased. These opposite effects (motor activation and sensory input suppression) were further discussed. PMID- 16473327 TI - Therapeutic action of cannabinoid on axonal injury induced by peroxynitrite. AB - This study examined whether the potent cannabinoid HU210 ameliorates axonal injury through its indirect action to stimulate the secretion of corticosterone. We observed that HU210 dramatically reduced peroxynitrite-induced axonal injury in rats receiving adrenalectomy and corticosterone replacement treatment. These results suggest that the ameliorating effects of cannabinoids on axonal injury associated with multiple sclerosis are achieved by its direct action, but not by its indirect action to elevate the serum corticosterone levels. PMID- 16473328 TI - Adenoviral gene transfer of BDNF and GDNF synergistically prevent motoneuron loss in the nucleus ambiguus. AB - We have previously shown that neuroprotective effects of an adenoviral glial cell line-derived neurotrophic factor (GDNF) gene transfer on the lesioned adult rat motoneurons in the nucleus ambiguus. In the present study, we examined neuroprotective effects of adenoviral gene transfer of brain-derived neurotrophic factor (BDNF) or/and GDNF to motoneurons in nucleus ambiguus using an adult rat vagal nerve avulsion model. The animals avulsed and inoculated with adenoviral vectors encoding BDNF (AxCAmBDNFME) or/and GDNF (AxCAhGDNF) showed immunolabeling for BDNF or/and GDNF in the nucleus ambiguus on the treated side, respectively, and expression of virus-induced BDNF or/and GDNF mRNA transcripts in the brainstem tissue that contained the nucleus ambiguus of the treated side. The treatment with AxCAhGDNF or AxCAmBDNFME significantly prevented the loss of vagal motoneurons in comparison to the control; the protective effect of AxCAmBDNFME was greater than that of AxCAhGDNF. The combined treatment with AxCAmBDNFME and AxCAhGDNF acted synergistically and significantly larger number of vagal motoneurons was preserved as compared to either AxCAmBDNFME treatment or AxCAhGDNF treatment. The treatment with AxCAmBDNFME or/and AxCAhGDNF after avulsion also suppressed the activity of nitric oxide synthase in lesioned motoneurons in the nucleus ambiguus. These results indicate that adenovirus mediated BDNF and GDNF gene transfer may prevent the degeneration of motoneurons in humans after either vagal nerve injury or recurrent laryngeal nerve injury. PMID- 16473329 TI - Neural correlates of within-level and across-level attention to multiple compound stimuli. AB - Event-related potentials (ERPs) were recorded to investigate the neural mechanisms of attention to the same or different levels of two compound letters presented concurrently in the left and right visual fields, respectively. Relative to the condition when attention was allocated to the global level of one compound stimulus and the local level of another one (across-level attention), attention to the same level of the two compound stimuli (within-level attention) increased an early positivity between 100 and 140 ms (P1) over the occipito parietal cortex. A long-latency positivity between 320 and 560 ms (P3) over the central-parietal area was also increased in the within-level relative to across level attention conditions. The ERP results suggest that, relative to across level attention, within-level attention to multiple compound stimuli facilitates both early sensory-perceptual processing and late process of stimulus evaluation and identification in hierarchical analysis. PMID- 16473330 TI - The pre-ischaemic neuroprotective effects of the polyamine analogues BU43b and BU36b in permanent and transient focal cerebral ischaemia models in mice. AB - The present study investigated the neuroprotective potential of two novel polyamine analogues, BU43b and BU36b, when administered 30 min prior to cerebral ischaemia. Neuroprotection in a permanent and a transient focal cerebral ischaemia mouse model (induced by intraluminal middle cerebral artery occlusion (MCAO)) was investigated using a range of histological and behavioural assessments. In the permanent ischaemia model, BU43b reduced oedema and showed a trend towards reduction in %HLV (percentage hemisphere lesion volume) when administered at a dose of 30 mg/kg i.p. Following transient ischaemia, treatment with BU43b decreased the %HLV and reduced oedema when administered at 30 mg/kg. BU43b also improved the locomotor activity (LMA) in MCAO mice at both 20 mg/kg and 30 mg/kg doses. BU36b was less effective than BU43b in both the permanent and the transient models, with its most pronounced effect being a trend towards reduction in oedema in both models. These results demonstrate that BU43b administered 30 min before ischaemia provided a good level of neuroprotection in the two models of cerebral ischaemia used and may have potential as a neuroprotective treatment for stroke. PMID- 16473331 TI - Adrenergic nerves mediate the venoconstrictor response to PVN stimulation. AB - Veins play an important role in the control of venous return, cardiac output and cardiovascular homeostasis. However, the central nervous system sites and effector systems involved in modulating venous function remain to be fully elucidated. The hypothalamic paraventricular nucleus (PVN) is an important site modulating autonomic outflow to the cardiovascular system. Venous tone can be modulated by sympathetic nerves or by adrenal catecholamines. The present study assessed the relative contribution of these autonomic effector systems to the venoconstrictor response elicited by stimulation of the hypothalamic paraventricular nucleus. Male Sprague-Dawley rats were subjected to sham operation or bilateral adrenal demedullation fitted with PVN guide cannulae and fitted with catheters for recording arterial pressure (AP) and intrathoracic vena caval pressure (VP). A latex balloon was advanced into the right atrium. MCFP was calculated from the AP and VP recorded after 4 s of right atrial occlusion. MCFP = VP + (AP - VP)/60. Mean arterial pressure (MAP), heart rate (HR), VP and MCFP responses to injections of BMI (25 ng/side) into the PVN were recorded from conscious rats to avoid the complicating effects of anesthesia. In sham-operated rats, injection of BMI into the PVN increased MAP by 13 +/- 3 mm Hg and HR by 56 +/- 6 bpm. MCFP was also increased significantly by 0.98 +/- 0.15 mm Hg indicating an increase in venomotor tone. Adrenal medullectomy did not affect the pressor (DeltaMAP = 12 +/- 2 mm Hg), tachycardic (DeltaHR = 48 +/- 7 bpm) or venoconstrictor (DeltaMCFP = 0.73 +/- 0.11 mm Hg) responses. Ganglionic blockade abolished the PVN-induced responses in both groups of rats. In a separate group, pretreatment with the adrenergic neuron blocker, guanethidine (20 mg/kg), also abolished the PVN-mediated venoconstrictor responses. Conversely, selective beta2 adrenergic receptor blockade did not affect MCFP responses to BMI. These data indicate that adrenomedullary catecholamines are not necessary for full expression of the venoconstrictor response to PVN stimulation. PMID- 16473332 TI - Isoflurane exerts neuroprotective actions at or near the time of severe traumatic brain injury. AB - Isoflurane improves outcome vs. fentanyl anesthesia, in experimental traumatic brain injury (TBI). We assessed the temporal profile of isoflurane neuroprotection and tested whether isoflurane confers benefit at the time of TBI. Adult, male rats were randomized to isoflurane (1%) or fentanyl (10 mcg/kg iv bolus then 50 mcg/kg/h) for 30 min pre-TBI. Anesthesia was discontinued, rats recovered to tail pinch, and TBI was delivered by controlled cortical impact. Immediately post-TBI, rats were randomized to 1 h of isoflurane, fentanyl, or no additional anesthesia, creating 6 anesthetic groups (isoflurane:isoflurane, isoflurane:fentanyl, isoflurane:none, fentanyl:isoflurane, fentanyl:fentanyl, fentanyl:none). Beam balance, beam walking, and Morris water maze (MWM) performances were assessed over post-trauma d1-20. Contusion volume and hippocampal survival were assessed on d21. Rats receiving isoflurane pre- and post-TBI exhibited better beam walking and MWM performances than rats treated with fentanyl pre- and any treatment post-TBI. All rats pretreated with isoflurane had better CA3 neuronal survival than rats receiving fentanyl pre- and post-TBI. In rats pretreated with fentanyl, post-traumatic isoflurane failed to affect function but improved CA3 neuronal survival vs. rats given fentanyl pre- and post-TBI. Post-traumatic isoflurane did not alter histopathological outcomes in rats pretreated with isoflurane. Rats receiving fentanyl pre- and post-TBI had the worst CA1 neuronal survival of all groups. Our data support isoflurane neuroprotection, even when used at the lowest feasible level before TBI (i.e., when discontinued with recovery to tail pinch immediately before injury). Investigators using isoflurane must consider its beneficial effects in the design and interpretation of experimental TBI research. PMID- 16473333 TI - The multiple dorsoventral origins and migratory pathway of tectal oligodendrocytes in the developing chick. AB - Oligodendrocytes have been considered to originate in a restricted ventricular zone of the ventral neural tube and to migrate and mature in their final targets. However, recent studies indicate that oligodendrocytes arise from multiple distinct dorsoventral origins. In this study, we investigate oligodendrocyte lineage cells in the embryonic optic tectum of chick, which develops from the dorsal region of the neural tube and invasion of optic tract. Oligodendrocyte precursor cells (OPCs) first appeared bilaterally on either side of the floor plate at E5. With further development, OPCs increased and spread laterally and dorsally to populate the optic tectum. At E7, OPCs appeared in another site along the ventral midline of the third ventricle, just dorsal to the optic chiasm. To examine the migration routes of these ventrally derived OPCs, we used DiI tracing in the organic culture and retinal denervation. Our results reveal that OPCs dispersed bilaterally along the optic tract and then migrated to the optic tectum in the stratum opticum (SO). In addition to these extrinsic OPCs, OPCs intrinsic to the tectal ventricle zone were identified at E14 using a combination of immunohistochemistry and retroviral mediated lineage tracing studies. These data support stage-specific dorsoventral origins and distribution of oligodendrocytes populating the optic tectum. PMID- 16473334 TI - Ibotenic acid lesions of the medial preoptic area disrupt the expression of partner preference in sexually receptive female rats. AB - The present study evaluated the effects of ibotenic acid lesions of the medial preoptic area (mPOA) on the display of partner preference in ovariectomized, estrogen- and progesterone-primed rats. Preference for a sexually vigorous male or an estrous female rat was determined in one of two conditions: unlimited physical access to the stimulus rats (Contact condition) or access that was limited to olfactory, auditory and visual cues (No-contact condition). Lesions of the mPOA reduced the male preference, social preference, and arena crossings, independent of test condition. However, the reduction in male preference following mPOA lesions was most pronounced during tests with unlimited physical access. These results suggest that the mPOA may be involved in integrating somatosensory signals from coital stimulation with the motor responses associated with the appetitive aspects of female sexual behavior. PMID- 16473335 TI - Interconnection between orexigenic neuropeptide Y- and anorexigenic alpha melanocyte stimulating hormone-synthesizing neuronal systems of the human hypothalamus. AB - Peripheral feeding-related hormones such as leptin, insulin, and ghrelin exert their main central effects through neuropeptide Y- (NPY) synthesizing and alpha melanocyte-stimulating hormone- (alpha-MSH) synthesizing neurons of the hypothalamic arcuate nucleus. In rodents, recent reports have described an asymmetric signaling between these neuron populations by showing that while NPY influences alpha-MSH-synthesizing neurons, the melanocortin-receptor agonist Melanotan II (MTII) does not modulate the electrophysiological properties of NPY neurons. The functional neuroanatomy of the relationship between these cell populations is unknown in humans. The aim of the current study was to analyze the putative relationship of the orexigenic NPY and anorexigenic alpha-MSH systems in the infundibular nucleus of the human hypothalamus, the analogue of the rodent arcuate nucleus. Double-labeling fluorescent immunocytochemistry for NPY and alpha-MSH was performed on postmortem sections of the human hypothalamus. The sections were analyzed by confocal laser microscopy. Both NPY- and alpha-MSH immunoreactive (IR) neurons were embedded in dense, intermingling networks of NPY and alpha-MSH-IR axons in the human infundibular nucleus. NPY-IR varicosities were observed in juxtaposition to all alpha-MSH-IR neurons. The mean number of NPY-IR axon varicosities on the surface of an alpha-MSH-IR neuron was approximately six. The majority of NPY-IR neurons were also contacted by alpha MSH-IR varicosities, although, the number of such contacts was lower (two alpha MSH-IR varicosities per NPY neuron). In summary, the present data demonstrate that these two antagonistic, feeding-related neuronal systems are interconnected in the infundibular nucleus, and the neuronal wiring possesses an asymmetric character in the human hypothalamus. PMID- 16473336 TI - Long-term memory for aversive training is impaired in Idua(-/-) mice, a genetic model of mucopolysaccharidosis type I. AB - Mucopolysaccharidosis type I (MPS I) is a lysosomal storage disease that leads to neurodegeneration and neurological deficits, among other pathological and clinical consequences. The aim of the present study was to evaluate neurobehavioral parameters in a genetic mouse model of mucopolysaccharidosis type I (MPS I). During exploration of an open field, adult MPS I (Idua(-/-)) mice showed normal locomotion and anxiety but reduced number of rearings. Idua(-/-) mice performed normally in a novel object recognition memory task and showed normal short-term retention of inhibitory avoidance training. By contrast, long term retention of inhibitory avoidance was impaired in Idua(-/-) mice. The deficit in inhibitory avoidance memory could not be attributed to reduced footshock reactivity. The results indicate that Idua(-/-) mice present deficits in long-term memory for aversive training and reduced exploratory behavior. PMID- 16473337 TI - Neurotrophic factors in the central nucleus of amygdala may be organized to provide substrates for associative learning. AB - The central nucleus of amygdala was examined to identify the ultrastructural distribution of neurotrophins responsible for the complex of neuronal signaling processes which regulate synaptic transmission and neuronal plasticity, and possibly underlie memory formation. We investigated at the electron microscopic level the cellular organization of brain-derived neurotrophic factor (BDNF) and its receptor, tyrosine kinase receptor B (TrkB), in the extended amygdala (CE). We also investigated the interaction between cortical inputs to CE and BDNF and TrkB. Our results indicate the presence of pro-BDNF and BDNF in terminals in the CE which show a strong association with immunoreactive postsynaptic densities. TrkB receptor immunoreactivity was localized to postsynaptic densities of asymmetric synapses on dendrites and dendritic spines. Cortical terminals formed asymmetric synapses with dendritic shafts and spines, but were not BDNF immunoreactive. TrkB receptors were observed opposed to cortical terminals. These data also suggest that one potential substrate for associative learning may be the interaction of different cortical inputs with neurotrophin-containing terminals ending on dendritic spines and other neuronal structures of CE. PMID- 16473338 TI - Visual motion and the neural correlates of event perception. AB - People perceive ongoing activity in terms of discrete temporal events. Distinctive changes in the movement of objects or actors may contribute to the perception that one event has ended and another has begun. However, little is known about the quantitative contribution of movement information to the processing of events. This study investigated how movement features are related to the neural processing of events by performing functional magnetic resonance imaging while participants viewed simple animations of moving objects. After the imaging session, participants watched the animations again and segmented them into meaningful events. Movement features were systematically related to viewers' perceptual segmentation and to cortical activity throughout visual processing areas. Activity in the MT complex, which is known to be specialized for processing motion, increased with increases in the objects' speed. The perception of an event boundary was associated with transient changes in the MT complex and in a nearby region in the superior temporal sulcus associated with processing biological motion. Other movement features were associated with changes in activity in occipital, parietal, and frontal cortex. These results indicate a role for movement features in the perceptual processing of meaningful events, and in the neural basis of that processing. PMID- 16473339 TI - Mentalizing and Marr: an information processing approach to the study of social cognition. AB - To interact successfully, individuals must not only recognize one another as intentional agents driven primarily by internal mental states, but also possess a system for making reliable and useful inferences about the nature of those beliefs, feelings, goals, and dispositions. The ability to make such mental state inferences (i.e., to mentalize or mindread) is the central accomplishment of human social cognition. The present article suggests that our understanding of how humans go about making mental state inferences will benefit from treating social cognition primarily as an information processing system that comprises a set of mechanisms for elaborating more basic social information into an understanding of another's mind. Following Marr's [Marr, D., 1982. Vision. W. H. Freeman, San Francisco, CA] framework for the study of such information processing systems, I suggest that questions about social cognition might profitably be asked at three levels--computation, algorithm, and implementation- and outline a number of ways in which a description of social cognition at the middle level (i.e., the step-by-step processes that give rise to mental state inferences) can be informed by analysis at the other two. PMID- 16473340 TI - Predicting Autism Spectrum Quotient (AQ) from the Systemizing Quotient-Revised (SQ-R) and Empathy Quotient (EQ). AB - BACKGROUND: Empathizing is a specific component of social cognition. Empathizing is also specifically impaired in autism spectrum condition (ASC). These are two dimensions, measurable using the Empathy Quotient (EQ) and the Autism Spectrum Quotient (AQ). ASC also involves strong systemizing, a dimension measured using the Systemizing Quotient (SQ). The present study examined the relationship between the EQ, AQ and SQ. The EQ and SQ have been used previously to test for sex differences in 5 'brain types' (Types S, E, B and extremes of Type S or E). Finally, people with ASC have been conceptualized as an extreme of the male brain. METHOD: We revised the SQ to avoid a traditionalist bias, thus producing the SQ-Revised (SQ-R). AQ and EQ were not modified. All 3 were administered online. SAMPLE: Students (723 males, 1038 females) were compared to a group of adults with ASC group (69 males, 56 females). AIMS: (1) To report scores from the SQ-R. (2) To test for SQ-R differences among students in the sciences vs. humanities. (3) To test if AQ can be predicted from EQ and SQ-R scores. (4) To test for sex differences on each of these in a typical sample, and for the absence of a sex difference in a sample with ASC if both males and females with ASC are hyper-masculinized. (5) To report percentages of males, females and people with an ASC who show each brain type. RESULTS: AQ score was successfully predicted from EQ and SQ-R scores. In the typical group, males scored significantly higher than females on the AQ and SQ-R, and lower on the EQ. The ASC group scored higher than sex-matched controls on the SQ-R, and showed no sex differences on any of the 3 measures. More than twice as many typical males as females were Type S, and more than twice as many typical females as males were Type E. The majority of adults with ASC were Extreme Type S, compared to 5% of typical males and 0.9% of typical females. The EQ had a weak negative correlation with the SQ-R. DISCUSSION: Empathizing is largely but not completely independent of systemizing. The weak but significant negative correlation may indicate a trade-off between them. ASC involves impaired empathizing alongside intact or superior systemizing. Future work should investigate the biological basis of these dimensions, and the small trade-off between them. PMID- 16473341 TI - On-line flexibility of the cognitive tuning of corticospinal excitability: a TMS study in human gait. AB - Human subjects have been found to be able to cognitively prepare themselves to resist to a TMS-induced central perturbation by selectively modulating the corticospinal excitability (CS). The aim of this study was to investigate the on line adaptability of this cognitive tuning of CS excitability during human gait. Transcranial magnetic stimulation (TMS) was used both as a central perturbation evoking a movement and as a tool for quantifying the CS excitability before the movement was evoked. TMS was applied at mid-stance (evoking additional hip extension) or at the beginning of the swing (evoking hip flexion) with a random phase, thus evoking unpredictable flexion or extension movement. This was compared to a condition of fixed phase, in which the subjects knew in advance the direction of the evoked movement. In both conditions, we compared the amplitude of the TMS-evoked movement and the motor-evoked potentials (MEPs) of the muscles acting at the hip joint (RF/BF) according to two opposite instructions, either to cognitively prepare to "let go", or to cognitively prepare to "compensate" for the evoked movements. The results showed that the subjects were able to compensate for random TMS-evoked movements, but with a lower performance level in comparison to the fixed TMS-evoked movements. When they succeeded in the random phase condition, the subjects used the same preparation strategy as in the fixed phase condition; preparing to compensate resulted in a selective increase in the CS excitability to those muscles which would be involved in counteracting the possible central perturbation. This requires continuous change in the tuning of CS excitability within the stride and thus reveals the high flexibility of the cognitive tuning of CS excitability during gait. PMID- 16473342 TI - Glycosidation of 2,5-anhydro-3,4-di-O-benzyl-D-mannitol with different glucopyranosyl donors: a comparative study. AB - 2,5-Anhydro-3,4-di-O-benzyl-D-mannitol was glycosylated using different donors such as tetra-O-acetyl-alpha-D-glucopyranosyl bromide in the presence of Hg(CN)(2), the corresponding beta-thiophenylglycoside in the presence of NIS and TfOH as well as the alpha- and beta-trichloroimidate with TMSOTf as promoter. The resulting mixtures were analyzed by HPLC and the following main components were isolated and characterized: 2,5-anhydro-3,4-di-O-benzyl-1-O-(2,3,4,6-tetra-O acetyl-beta-D-glucopyranosyl)-d-mannitol; 6-O-acetyl-2,5-anhydro-3,4-di-O-benzyl 1-O-(2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl)-D-mannitol; 2,5-anhydro-3,4-di O-benzyl-1,6-bis-O-(2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl)-D-mannitol; 2,5 anhydro-3,4-di-O-benzyl-1-O-[-2-O-(2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl) 3,4,6-tri-O-acetyl-beta-D-glucopyranosyl]-6-O-(2,3,4,6-tetra-O-acetyl-beta-D glucopyranosyl)-D-mannitol and 2,5-anhydro-3,4-di-O-benzyl-1,6-bis-O-(3,4,6-tri-O acetyl-1,2-O-ethylidene-2'-yl-alpha-D-glucopyranosyl)-D-mannitol. The latter compound representing a bis-orthoester might be a common intermediate in all the investigated reactions, as its rearrangement and/or decomposition can yield all of the isolated compounds. PMID- 16473343 TI - Evaluation of essential and toxic metals by ultrasound-assisted acid leaching from scalp hair samples of children with macular degeneration patients. AB - BACKGROUND: The causes of night blindness in children are multifactorial and particular consideration has been given to childhood nutritional deficiency, which is the most common problem found in underdeveloped countries. Such deficiency can result in physiological and pathological processes that in turn influence hair composition. METHOD: An ultrasonic-assisted acid leaching procedure was developed as a sample pretreatment for the determination of Zn, Cu, Cd, As and Pb in human scalp hair samples of night blindness male children with age between 5 to 15 y and compared with the children without vision anomalies that lived in the same localities. The effects of different factors on acid leaching of metals, such as preintensification time (without ultrasonic stirring) after treatment of acid mixture, exposure time to ultrasound and temperature of the ultrasonic bath have been investigated. The proposed method was validated by certified reference samples of scalp hair CRM 397. The wet acid digestion method was used to obtain the total metal concentration in both scalp hair and CRM samples. Cu and Zn in leachates and digests were measured by flame atomic absorption spectrometry (FAAS) using a conventional air/acetylene flame, while Cd and Pb were determined by electrothermal atomic absorption spectrometry (ETAAS) under optimized conditions. RESULTS: It was observed that at optimal conditions, the recovery for Zn, Cd, Pb, As and Cu were 98%, 98.5%, 96%, 97.2% and 94% respectively. The mean values of Zn and Cu in scalp hair samples of children having night blindness were significantly lower as compared to normal healthy children (p for Zn<0.001 and Cu<0.003), while the level of toxic metals As, Cd and Pb were significantly higher in children having ocular problems as related to normal children (p As<0.0074, Cd<0.001 and lead<0.004). CONCLUSION: These data present guidance to clinicians and other professional investigating deficiency of essential trace metals and excessive level of toxic metals in biological samples. PMID- 16473344 TI - Ultrasonic backscattering in tissue: characterization through Nakagami generalized inverse Gaussian distribution. AB - Ultrasonic tissue characterization through composite probability distributions such as Nakagami-lognormal, Nakagami-gamma, Nakagami-inverse Gaussian has been found to be useful. Such a probabilistic description also depicts heavy tails which arise from multiple scattering in tissue besides local and global variations in scattering cross-sections. A new composite probability distribution, viz. Nakagami-generalized inverse Gaussian distribution (NGIGD) with four parameters is proposed which under different limiting conditions results in approximating the known distributions. A salient aspect of the new distribution is that the probability density function (pdf) of NGIGD variate is available in closed form and is analytically tractable. PMID- 16473345 TI - The histidine kinase homologue DhkK/Sombrero controls morphogenesis in Dictyostelium. AB - A key event in Dictyostelium development is the formation of the Mexican hat. This corresponds to a commitment step in morphogenesis that irreversibly signals progression from the slug stage to the fruiting body. We describe the characterization of the dhkK gene that controls this morphogenetic step. Null mutants of dhkK repeatedly attempt, and fail, to undergo morphogenesis from the slug to the Mexican hat, causing them to exhibit a "slugger" phenotype, which cannot be corrected by co-development with wild-type cells. The dhkK gene encodes a putative receptor histidine kinase whose expression is enriched in prestalk cells in the slug. Uniquely for a histidine kinase, DhkK is located in the nuclear envelope. Entry into culmination requires the DhkK response regulator domain, which appears to directly regulate cyclic AMP signaling. PMID- 16473347 TI - The influence of the time course of inflammation and spinalization on the antinociceptive activity of the alpha2-adrenoceptor agonist medetomidine. AB - The purpose of the present study was to investigate the influence of the time course of inflammation and the implication of spinal and supraspinal sites on the antihyperalgesic effects of the alpha(2)-adrenoceptor agonist medetomidine. Behavioral experiments showed a more intense antihyperalgesia in the phase of maintenance of inflammation than in the early or resolution stages. Maximum effect, without sedation, was observed with a dose of 40 microg/kg (66+/-12% and 76+/-15% reduction of mechanical and thermal hyperalgesia). No change was observed in the paw swelling, indicating that its effects were not secondary to a reduction of inflammation. In electrophysiological experiments, the effect was more pronounced in animals with an intact spinal cord than in spinalized animals (max. effects of 2+/-0.7% vs. 48+/-11% of control, noxious mechanical stimulation). We conclude that the antihyperalgesic effect of medetomidine depends on the time course of inflammation and that it is mainly located supraspinally. PMID- 16473346 TI - Novel concepts of neuropeptide-based drug therapy: vasoactive intestinal polypeptide and its receptors. AB - Chronic inflammatory airway diseases such as bronchial asthma or chronic obstructive pulmonary disease (COPD) are major contributors to the global burden of disease. Although inflammatory cells play the central role in the pathogenesis of the diseases, recent observations indicate that also resident respiratory cells represent important targets for pulmonary drug development. Especially targeting airway neuromediators offers a possible mechanism by which respiratory diseases may be treated in the future. Among numerous peptide mediators such as tachykinins, calcitonin gene-related peptide, neurotrophins or opioids, vasoactive intestinal polypeptide (VIP) is one of the most abundant molecules found in the respiratory tract. In human airways, it influences many respiratory functions via the receptors VPAC1, VPAC2 and PAC1. VIP-expressing nerve fibers are present in the tracheobronchial smooth muscle layer, submucosal glands and in the walls of pulmonary and bronchial arteries and veins. Next to its strong bronchodilator effects, VIP potently relaxes pulmonary vessels, and plays a pivotal role in the mediation of immune mechanisms. A therapy utilizing the respiratory effects of VIP would offer potential benefits in the treatment of obstructive and inflammatory diseases and long acting VIP-based synthetic non peptide compounds may represent a novel target for drug development. PMID- 16473348 TI - Class Ic antiarrhythmics block human skeletal muscle Na channel during myotonia like stimulation. AB - Flecainide, a class Ic antiarrhythmic drug, has been anecdotally reported to improve myotonia, but little is known about its kinetics on human skeletal muscle sodium channels applicable in vivo. Here we explored the anti-myotonic action of flecainide for human skeletal muscle sodium channels heterologously expressed in cultured cells. Flecainide blocked sodium channels in a highly state-dependent manner with 20-fold difference in IC(50) between use-dependent and tonic blocks. When pulses of brief depolarization simulating myotonia were applied from a holding potential of -90 mV, flecainide at therapeutic concentrations significantly blocked sodium currents. Flecainide slowed the time course of recovery but most channels recovered from block within 10-20 s. In contrast to mexiletine, flecainide did not markedly block sodium current during prolonged depolarization, suggesting an open-channel blocking action. Considering the slow recovery from block and the specific action against repetitive depolarization, flecainide may represent a potent therapeutic agent for myotonia. PMID- 16473349 TI - Membrane-type 1 matrix metalloproteinase modulates focal adhesion stability and cell migration. AB - Membrane-type 1 matrix metalloproteinase (MT1-MMP) plays an important role in extracellular matrix-induced cell migration and the activation of extracellular signal-regulated kinase (ERK). We showed here that transfection of the MT1-MMP gene into HeLa cells promoted fibronectin-induced cell migration, which was accompanied by fibronectin degradation and reduction of stable focal adhesions, which function as anchors for actin-stress fibers. MT1-MMP expression attenuated integrin clustering that was induced by adhesion of cells to fibronectin. The attenuation of integrin clustering was abrogated by MT1-MMP inhibition with a synthetic MMP inhibitor, BB94. When cultured on fibronectin, HT1080 cells, which endogenously express MT1-MMP, showed so-called motile morphology with well organized focal adhesion formation, well-oriented actin-stress fiber formation, and the lysis of fibronectin through trails of cell migration. Inhibition of endogenous MT1-MMP by BB94 treatment or expression of the MT1-MMP carboxyl terminal domain, which negatively regulates MT1-MMP activity, resulted in the suppression of fibronectin lysis and cell migration. BB94 treatment promoted stable focal adhesion formation concomitant with enhanced phosphorylation of tyrosine 397 of focal adhesion kinase (FAK) and reduced ERK activation. These results suggest that lysis of the extracellular matrix by MT1-MMP promotes focal adhesion turnover and subsequent ERK activation, which in turn stimulates cell migration. PMID- 16473350 TI - Identification of novel genes regulated in the developing human ventral mesencephalon. AB - In the human embryo, from approximately 6 weeks gestational age (GA), dopaminergic (DA) neurons can be found in the ventral mesencephalon (VM). More specifically, the post-mitotic neurons are located in the ventral part of the tegmentum (VT), whereas no mature DA neurons are found in the neighboring dorsal part. We used Affymetrix HG-U133 GeneChip technology to compare genome-wide expression profiles of ventral and dorsal tegmentum from 8 weeks GA human embryos, in order to identify genes involved in specification, differentiation, and survival of mesencephalic DA (mDA) neurons. Known mDA marker genes including ALDH1A1, DAT1, VMAT2, TH, CALB1, NURR1, FOXA1, GIRK2, PITX3, RET, and DRD2 topped the list of 96 genes from HG-U133A with higher expression in VT, validating the experimental set-up. In addition, 28 probes from HG-U133B were identified whereof most are annotated to UniGene clusters with no gene associated or to genes of unknown function. Of these, the fifteen most regulated transcripts, representing changes down to 56% could be verified by quantitative real-time PCR (Q-PCR) on a developmental series of subdissected human embryonic and fetal brain material, resulting in not only a regional but also a temporal expression profile. This revealed a distinct DA-associated profile for in particular a putative transcription factor (FLJ45455) and the uncharacterized transmembrane proteins KIAA1145 and SLC10A4. The data presented here may help to device cell replacement and regenerative therapies for Parkinson's disease (PD). PMID- 16473351 TI - The effects of metronidazole and furazolidone during Giardia differentiation into cysts. AB - The protozoon Giardia lamblia infects millions of people worldwide, most of them in underdeveloped countries, where it is frequently a hyperendemic disease. The search for an effective anti-Giardia treatment has been intense, but recurrent infections, virulence factors, and drug resistance imposed obstacles in the achievement of an efficient medication. Most papers about drug effects in Giardia are related to the trophozoite form, although viable cysts, the infective forms, are continuously eliminated in the stools during the treatment. Supported by this knowledge, we analyzed the inhibitory effects of metronidazole (MZ) and furazolidone (FZ) on the differentiation of Giardia into cysts and its viability. The presence of cavities, lamellar bodies and thread-like structures were the most frequent morphological alterations. The results showed also that FZ was more effective by 50% than MZ in inhibiting in vitro cyst differentiation. PMID- 16473352 TI - Sex steroid concentrations and localization of steroidogenic enzyme expression in free-ranging female northern fur seals (Callorhinus ursinus). AB - Recent precipitous population declines in northern fur seal (Callorhinus ursinus) and other Alaskan pinniped populations are due in part to reduced fecundity and have emphasized deficits in basic reproductive knowledge of these species. Following estrus and mating, fertilized female pinnipeds experience an obligatory delayed implantation lasting several months and non-pregnant (pseudopregnant) females are indistinguishable by sex steroid levels during this time. The current study examined circulating steroid concentrations and ovarian expression of key steroid-synthesizing enzymes in northern fur seals to identify changes associated with embryonic implantation, data necessary for estimating early pregnancy rates of the population. Blood samples were collected from 84 female fur seals captured on an Alaskan rookery from October 15 to November 30, a period spanning the end of the delay and subsequent uterine implantation of the fertilized blastocyst in this species. Concentrations of progesterone, estradiol, and estrone measured in blood collected during boreal fall were contrasted with samples collected during late summer from post-ovulatory females and males. Serum sex steroids were also related to ovarian expression of key enzymes responsible for androgen (17alpha hydroxylase/17,20-lyase, cytochrome b5, and 3beta-hydroxysteroid dehydrogenase) and estrogen (aromatase cytochrome P450) synthesis. Enzymes necessary for androgen synthesis were highly expressed in ovaries, and accordingly, endocrine profiles were expanded to include DHEA, androstenediol, androstenedione, and testosterone. Estradiol concentrations were universally low and free and conjugated estrone were the primary circulating estrogens in fur seal sera. No implantation-associated peak was identified for estrogens or progesterone and mean values of progesterone and estrone were actually greater in female serum samples collected during summer than fall. However, there was a significant positive relationship between fall sampling date and testosterone concentration. Additionally, DHEA concentrations in females were lower in samples collected during the implantation period than during the summer embryonic delay. These data suggest androgens may play a substantial regulatory role in the embryonic delay of northern fur seals. PMID- 16473353 TI - Male 11-ketotestosterone levels change as a result of being watched in Siamese fighting fish, Betta splendens. AB - This study investigated the effects of nesting status and the presence of an audience on 11-ketotestosterone (11KT) levels in male Siamese fighting fish, Betta splendens. Prior studies have demonstrated that both nesting status, an indicator of territory-holding power and reproductive state, and the sex of a conspecific audience lead to differences in male behavior during aggressive encounters. Since behavioral changes have already been demonstrated, we chose to investigate whether 11KT levels were also influenced by nesting status and audience presence as 11KT both stimulates, and is stimulated by, reproductive and aggressive behaviors in male teleosts. Male 11KT levels were measured from water samples taken from containers holding fish both before and after interaction. Males interacted under three treatment conditions: no audience, female audience, and male audience. Within these treatments were two nest paradigms: both males had nests or neither male had a nest. 11KT levels varied depending on nesting status and audience type. In general, 11KT levels were lower in interacting males when a female audience was present or when males had nests. Overall, 11KT showed increases or decreases as aggression increased or decreased, as shown by already established behavioral findings [see Dzieweczynski T.L., Green T.M., Earley R.L., Rowland W.J., 2005. Audience effect is context dependent in Siamese fighting fish, Betta splendens. Behav. Ecol. 16, 1025-1030; Doutrelant, C., McGregor, P.K., Oliveira, R.F., 2001. Effect of an audience on intrasexual communication in male Siamese fighting fish (Betta splendens). Behav. Ecol. 12, 283-286.]. Our results suggest that 11KT levels are influenced by reproductive status, as indicated by nest ownership, and audience presence and are most likely modulated by territorial behavior and social environment. PMID- 16473354 TI - Differential effect of testosterone and repetitive induction on cataleptic and dorsal immobility in mice. AB - In nature, many species under conditions of stress (e.g., predator attack, pups carried by the mother, mating) show immobility states called "immobility responses" (IRs), which are characterized by the complete absence of movement and a relative unresponsiveness. These IR states can be induced by several kinds of sensorial stimuli. Many brain neurotransmitters from diverse cerebral areas participate in the expression of IRs. Other factors are also involved in IRs, such as learning and hormones, but at present, there is not enough experimental support about these factors. Our purpose was to investigate whether the IRs are subject to sexual hormone modulation and to examine the possible relation to learning processes. We tested the effects of acute testosterone decanoate (30 mg/kg, s.c.) and repetitive induction of two IRs; cataleptic immobility (CAT) and dorsal immobility (DI). These were tested in mice of both sexes which were either gonadectomized or sham-treated. CAT and DI were measured before and then 1 and 5 h after testosterone injection. The results show a differential effect of the repetitive induction on CAT and DI. CAT was augmented with repetition, and DI was decreased. Sex differences of the effects of the acute testosterone treatment were observed. Sham and castrated male mice showed CAT potentiation; in contrast, DI was reduced albeit only in sham male mice. Sham and ovariectomized female mice were not affected by testosterone. These results support the hypothesis that there are multiple immobility systems that can be differentially modulated by brain regions associated with processes of learning. PMID- 16473355 TI - Common errors in the design of orthopaedic studies. AB - There are several issues that one has to take into consideration in order to avoid potential pitfalls in the design of orthopaedic studies. This article highlights how to avoid common errors and how to continue the drive towards the unattainable, but laudable, goal of perfection. PMID- 16473356 TI - Conducting industry-partnered trials in orthopaedic surgery. AB - We have seen an emergence of larger scale collaborative multi-center trials in surgery. These larger trials have the advantage of increased generalisability of the results and the potential for large scale and efficient recruitment (1000 patients or more). It is estimated that the average cost of bringing a new drug to market approximates 500 million dollars. The funding limitations in most national peer-reviewed agencies provides a compelling rationale for industry co sponsors. To limit biases associated with industry-sponsored research, researchers should develop specific protocols to ensure accurate and transparent reporting of funding sources, design and implementation of surgical trials, manuscript preparation and the criteria for authorship. PMID- 16473357 TI - Integrating 'mental illness' and 'motherhood': the positive use of surveillance by health professionals. A qualitative study. AB - BACKGROUND: Sociologists have long recognised the social control functions of different social institutions. Nurses, however, often appear more comfortable with formulating their roles in altruistic terms. OBJECTIVES: In this paper, we examine the relevance of Foucauldian concepts, in particular that of surveillance, to an understanding the relationship between healthcare professionals (especially nurses) and their patients. DESIGN: We use the concept of 'interactional frames' to analyse data from qualitative interviews with mothers who have a mental illness. SETTINGS: The research, from which the data in this paper were taken, was carried out in a largely urban area of south-east Wales, in the UK, during 2001 and 2002. PARTICIPANTS: The participants were 11 women, each with one or more children, all of whom were under the care of their local Community Mental Health Team. METHODS: The paper draws on findings from a wider study of the influence of child-care responsibilities on access to services for women with mental health problems. Data were generated through individual, semi-structured interviews, carried out and transcribed by one of the authors (BD). RESULTS: Women produced accounts of their mothering practices which acknowledged the norms of 'good' mothering. They spoke about the need for 'impression management' in their clinical encounters, both those in which they were the patient and those undertaken on behalf of their children. The data showed health professionals moving between frames in which the woman was a mother and in which she was a person with a mental illness, and integrating the two frames to the woman's benefit. CONCLUSIONS: Women who are mothers and who are also users of mental health services face particular challenges in managing the contradictory aspects of their dual identity. Health professionals can use their disciplinary power in a positive way, to help women in this task. PMID- 16473358 TI - The cis-9,trans-11 isomer of conjugated linoleic acid (CLA) lowers plasma triglyceride and raises HDL cholesterol concentrations but does not suppress aortic atherosclerosis in diabetic apoE-deficient mice. AB - OBJECTIVE: Reduction in atherosclerosis has been reported in experimental animals fed mixtures of conjugated linoleic acid (CLA). In this study, the major naturally occurring CLA isomer (cis-9,trans-11) was tested in an atherosclerosis prone mouse model. METHODS: In a model of insulin deficient apoE deficient mice, 16 animals were fed for 20 weeks with supplemental CLA (09.%, w/w) and compared with a similar number of mice of this phenotype. A control comparison was made of metabolic changes in non-diabetic apoE deficient mice that develop little atherosclerosis over 20 weeks. At 20 weeks, plasma lipids were measured and aortic atherosclerosis quantified by Sudan staining in the arch, thoracic and abdominal segments. RESULTS: The diabetic apoE deficient mice developed marked dyslipidemia, primarily as cholesterol-enriched chylomicron and VLDL-sized lipoproteins and atherosclerosis in the aortic arch. However, there were no significant differences between CLA fed and non-CLA fed mice in either phenotype in plasma cholesterol concentration (in diabetic: 29.4+/-7.7 and 29.5+/-5.9 mmol/L, respectively) or in the area of aortic arch atherosclerosis (in diabetic: 24.8+/-10.3 and 27.6+/-7.7%, respectively). However, among diabetic mice the triglyceride concentration in triglyceride-rich lipoproteins was significantly lower in those fed CLA (for plasma 2.2+/-0.8 to 1.1+/-0.3 mmol/L; P<0.001), a significant difference that was seen also in the non-diabetic mice in which HDL cholesterol increased significantly with CLA (0.35+/-0.12-0.56+/-0.15 mmol/L). CONCLUSION: In this atherosclerosis-prone model, the diabetic apoE deficient mouse, supplemental 0.9% CLA (cis-9,trans-11) failed to reduce the severity of aortic atherosclerosis, although plasma triglyceride concentration was substantially lowered and HDL cholesterol raised. PMID- 16473359 TI - Theoretical investigations of the chromatographic separation of interacting enantiomers. AB - Separation of a pair of enantiomers by liquid chromatography is modeled using the equilibrium dispersive (ED) model of chromatography. It is assumed that the chiral stationary phase used for the separation consists of two types of adsorption sites, including chiral selectors linked to the surface and nonselective centers belonging to the achiral matrix. Additionally, intermolecular interactions between adsorbed enantiomers are taken into account. The corresponding equilibrium adsorption isotherms of the enantiomers are derived by means of the mean field approximation (MFA) and used as input data for the ED model. Special attention is paid to the influence of the lateral interactions on the effectiveness of the enantiomer separation. In particular, we examine the effect of the interactions on the shape and relative position of the chromatographic peaks associated with the enantiomers. Furthermore, the influence of the spacer length, which modifies screening of the lateral interactions, on the adsorption process is studied. The obtained results suggest that the lateral interactions combined with the screening effect may cause serious changes in the separation, depending on the nature (attraction or repulsion) and strength of the interactions as well as on the spacer length. PMID- 16473360 TI - Development of capillary column packed with thiol-modified gold-coated polystyrene particles and its selectivity for aromatic compounds. AB - Three types of thiol compounds (n-octadecanethiol, thiophenol, and 2 phenylethanethiol) were used to modify the gold-coated polystyrene particles (dp. 5microm) to prepare a stationary phase for capillary liquid chromatography through the formation of self-assembled monolayer. The column with n octadecanethiol-modified gold-coated polystyrene particles (C18-Au) demonstrated the higher affinity to phenanthrene and anthracene than small aromatics compared to the ODS column. In addition, the shape selectivity between phenanthrene and anthracene in the C18-Au column was much higher than that in the ODS column (separation factors: 1.82 and 1.14, respectively). The relationship between the retention factor and acetonitrile content in the mobile phase revealed that the retention behaviors in the C18-Au column was more sensitive on the acetonitrile content than those in the ODS column. Relatively higher affinity for phenanthrene and anthracene was commonly observed in all the three thiol-modified Au columns than that for the conventional ODS column, whereas separations of benzene and nitro- and chlorobenzenes were quite different among the three thiol-modified Au and ODS columns. PMID- 16473362 TI - Peak compression in reversed-phase gradient elution. AB - Previous reports suggest that peak widths in linear gradient elution are consistently larger than predicted by theory; however, if gradient compression is ignored, experiment and theory are in reasonable agreement. This suggests that gradient compression might represent an incorrect or poorly understood concept. In the present study, an experimental program was carried out to better understand the role of gradient compression and the reason for past differences between experiment and theory. It is concluded that the concept of gradient compression is correct. PMID- 16473361 TI - Method development for the determination of residual fluorotelomer raw materials and perflurooctanoate in fluorotelomer-based products by gas chromatography and liquid chromatography mass spectrometry. AB - The methodology for the determination of perfluorooctanoate (C(7)F(15)COO-, PFO), fluorotelomer alcohols (FTOHs: 6-2, 8-2, and 10-2), perfluorooctyl iodide (PFOI), and 8-2-8 fluorotelomer alcohol ester in complex fluorotelomer-based commercial products has been demonstrated and validated. Sample preparation procedures allowing determination of residual levels of these compounds were developed. The analytes were detected either by LC/MS/MS (PFO), LC/MS (FTOHs), or GC/MS (PFOI, 8 2-8 ester). The methods were validated by investigating the recoveries of analytes spiked at multiple levels to authentic sample matrices. The recoveries generally were between 70 and 130%. The limits of detection were in sub-microg/g range and the limits of quantitation were in the mug/g range. The methods were applied to fluorotelomer-based raw materials and fluorotelomer-based surfactants and polymeric products and represent methods useful for the determination of higher carbon chain length homologs as well. PMID- 16473363 TI - Stabilizer-free nanosized gold sols. AB - The paper describes a convenient, rapid, and reproducible method for the synthesis of stable dispersions of uniform gold nanoparticles at ambient temperatures by mixing aqueous solutions of tetrachloroauric acid and iso ascorbic acid. The influence of the experimental conditions on the size of the gold particles and the stability of the final sols was monitored by dynamic light scattering and UV-vis spectrophotometry. It was found that the size of the resulting nanoparticles is affected by the concentration and the pH of gold solution, while the stability of the electrostatically stabilized final sols is strongly dependent on the excess of reductant in the system, the ionic strength, and the temperature of the precipitation. Since the preparation process does not require the addition of a dispersing agent, the surface of the resulting gold nanoparticles can be easily functionalized to make them suitable for applications in medicine, biology, and catalysis. PMID- 16473364 TI - Effects of cleaning procedures of silica wafers on their friction characteristics. AB - Silicon wafers with thermal silicon oxide layers were cleaned and hydrophilized by three different methods: (1) the remote chemical analysis (RCA) wet cleaning by use of ammonia and hydrogen peroxide mixture solutions, (2) water-vapor plasma cleaning, and (3) UV/ozone combined cleaning. All procedures were found to remove effectively organic contaminations on wafers and gave identical characteristics of the contact angle, the surface roughness and the normal force interactions, measured by atomic force microscopy (AFM). However, it is found that wafers cleaned by the RCA method have several times larger friction coefficients than those cleaned by the plasma and UV/ozone methods. The difference was explained by the atomic-scale topological difference induced during the RCA cleaning. This study reveals the lateral force microscopy as a very sensitive method to detect the microstructure of surfaces. PMID- 16473365 TI - Short-term study of the uptake of PrP(Sc) by the Peyer's patches in hamsters after oral exposure to scrapie. AB - The disease-associated prion protein (PrP(Sc)) has been detected in the ileal Peyer's patches of lambs as early as one week after oral exposure to scrapie. In hamsters, the earliest reported time of PrP(Sc) detection in the Peyer's patches after oral exposure to scrapie is 69 days post-infection. To evaluate the acute uptake of inoculum and to investigate whether the Peyer's patches constitute the primary site of entry for scrapie after oral exposure, hamsters were each exposed orally to 1 ml of a 10% brain homogenate from hamsters in the terminal stage of infection with the 263 K strain of the scrapie agent. PrP(Sc) was demonstrated in the Peyer's patches only a few days after exposure, i.e., much earlier than previously reported. This study supports the view that the Peyer's patches constitute at least one of the primary entry sites of PrP(Sc) after oral exposure to scrapie. PMID- 16473366 TI - Analysis of the secondary structure of expansion segment 39 in ribosomes from fungi, plants and mammals. AB - The structure of expansion segment 39, ES39, in eukaryotic 23 S-like ribosomal RNA was analysed using a combination of chemical and enzymic reagents. Ribosomes were isolated from yeast, wheat, mouse, rat and rabbit, five organisms representing three different eukaryotic kingdoms. The isolated ribosomes were treated with structure-sensitive chemical and enzymic reagents and the modification patterns analysed by primer extension and gel electrophoresis on an ABI 377 automated DNA sequencer. The expansion segment was relatively accessible to modification by both enzymic and chemical probes, suggesting that ES39 was exposed on the surface of the ribosomes. The collected modification data were used in secondary structure modelling of the expansion segment. Despite considerable variation in both sequence and length between organisms from different kingdoms, the structure analysis of the expansion segment gave rise to structural fingerprints that allowed identification of homologous structures in ES39 from fungi, plants and mammals. The homologous structures formed an initial helix and an invariant hairpin connected to the initial helix via a long single stranded loop. The remaining part of the ES39 sequences accounted for most of the length variation seen between the analysed species. This part could form additional, albeit less similar, hairpins. A comparison of ES39 sequences from other fungi, plants and mammals showed that identical structures could be formed in these organisms. PMID- 16473367 TI - Structural characterization of an equilibrium unfolding intermediate in cytochrome c. AB - Although the denaturant-induced unfolding transition of cytochrome c was initially thought to be a cooperative process, recent spectroscopic studies have shown deviations from two-state behavior consistent with accumulation of an equilibrium intermediate. However, little is known about the structural and thermodynamic properties of this state, and whether it is stabilized by the presence of non-native heme ligands. We monitored the reversible denaturant induced unfolding equilibrium of oxidized horse cytochrome c using various spectroscopic probes, including fluorescence, near and far-UV CD, heme absorbance bands in the Soret, visible and near-IR regions of the spectrum, as well as 2D NMR. Global fitting techniques were used for a quantitative interpretation of the results in terms of a three-state model, which enabled us to determine the intrinsic spectroscopic properties of the intermediate. A well-populated intermediate was observed in equilibrium experiments at pH 5 using either guanidine-HCl or urea as a denaturant, both for wild-type cytochrome c as well as an H33N mutant chosen to prevent formation of non-native His-heme ligation. For a more detailed structural characterization of the intermediate, we used 2D 1H-15N correlation spectroscopy to follow the changes in peak intensity for individual backbone amide groups. The equilibrium state observed in our optical and NMR studies contains many native-like structural features, including a well structured alpha-helical sub-domain, a short Trp59-heme distance and solvent shielded heme environment, but lacks the native Met80 sulfur-iron linkage and shows major perturbations in side-chain packing and other tertiary interactions. These structural properties are reminiscent of the A-state of cytochrome c, a compact denatured form found under acidic high-salt conditions, as well as a kinetic intermediate populated at a late stage of folding. The denaturant-induced intermediate also resembles alkaline forms of cytochrome c with altered heme ligation, suggesting that disruption of the native methionine ligand favors accumulation of structurally analogous states both in the presence and absence of non-native ligands. PMID- 16473368 TI - Recapitulation and design of protein binding peptide structures and sequences. AB - An important objective of computational protein design is the generation of high affinity peptide inhibitors of protein-peptide interactions, both as a precursor to the development of therapeutics aimed at disrupting disease causing complexes, and as a tool to aid investigators in understanding the role of specific complexes in the cell. We have developed a computational approach to increase the affinity of a protein-peptide complex by designing N or C-terminal extensions which interact with the protein outside the canonical peptide binding pocket. In a first in silico test, we show that by simultaneously optimizing the sequence and structure of three to nine residue peptide extensions starting from short (1 6 residue) peptide stubs in the binding pocket of a peptide binding protein, the approach can recover both the conformations and the sequences of known binding peptides. Comparison with phage display and other experimental data suggests that the peptide extension approach recapitulates naturally occurring peptide binding specificity better than fixed backbone design, and that it should be useful for predicting peptide binding specificities from crystal structures. We then experimentally test the approach by designing extensions for p53 and dystroglycan based peptides predicted to bind with increased affinity to the Mdm2 oncoprotein and to dystrophin, respectively. The measured increases in affinity are modest, revealing some limitations of the method. Based on these in silico and experimental results, we discuss future applications of the approach to the prediction and design of protein-peptide interactions. PMID- 16473369 TI - Elongated oligomers assemble into mammalian PrP amyloid fibrils. AB - In prion diseases, the mammalian prion protein PrP is converted from a monomeric, mainly alpha-helical state into beta-rich amyloid fibrils. To examine the structure of the misfolded state, amyloid fibrils were grown from a beta form of recombinant mouse PrP (residues 91-231). The beta-PrP precursors assembled slowly into amyloid fibrils with an overall helical twist. The fibrils exhibit immunological reactivity similar to that of ex vivo PrP Sc. Using electron microscopy and image processing, we obtained three-dimensional density maps of two forms of PrP fibrils with slightly different twists. They reveal two intertwined protofilaments with a subunit repeat of approximately 60 A. The repeating unit along each protofilament can be accounted for by elongated oligomers of PrP, suggesting a hierarchical assembly mechanism for the fibrils. The structure reveals flexible crossbridges between the two protofilaments, and subunit contacts along the protofilaments that are likely to reflect specific features of the PrP sequence, in addition to the generic, cross-beta amyloid fold. PMID- 16473370 TI - Absence of cerebrospinal fluid oligoclonal bands is associated with delayed disability progression in relapsing-remitting MS patients treated with interferon beta. AB - To assess the role of CSF oligoclonal bands (OB) in determining the clinical outcome in patients with relapsing-remitting multiple sclerosis (RRMS) treated with IFN-beta, we carried out a retrospective, multicentre, observational study recruiting 209 RRMS patients from six MS centres from northern, central and southern areas of Italy under treatment with IFN-beta-1a i.m., IFN-beta-1a s.c. and IFN-beta-1b s.c. Twenty-two of 209 patients (10.6%) showed no OB in CSF. The patients without had, at disease onset, significantly higher frequency of visual disturbances (p=0.02) and less sensory involvement (p=0.04) than those with OB. A statistical trend (p=0.056) towards a longer time to reach sustained disability progression during treatment was found in patients without compared to those with OB. Thirty-six of 187 (19%) patients with OB worsened by at least 1 EDSS point compared to none of 22 (0%) OB-negative patients (p=0.017). The delaying of disability progression in OB-negative patients during treatment was significantly dependent only on the number of baseline MRI T2-weighted lesions (p=0.012) that was found to be significantly lower in OB-negative than in OB-positive patients (p=0.04). The absence of OB and low number of baseline T2-weighted lesions in this cohort of MS patients are favourable prognostic factors influencing the clinical response to IFN-beta treatment in RRMS patients. PMID- 16473371 TI - Differential patterns of dopamine transporter loss in the basal ganglia of progressive supranuclear palsy and Parkinson's disease: analysis with [(123)I]IPT single photon emission computed tomography. AB - We evaluated the patterns of dopamine transporter loss in the striatum of ten controls, twenty patients with Parkinson's disease (PD), and nine with progressive supranuclear palsy (PSP) using (123)I-IPT single photon emission tomography (SPECT). Four ROIs in the striatum correspond to the head of caudate nucleus (ROI 1), a transitional region between head of caudate and putamen (ROI 2), anterior putamen (ROI 3), and posterior putamen (ROI 4). A striatal ratio of specific to nondisplaceable uptake (V3'') was calculated normalizing the activity of the ROIs to that of occipital cortex. V3'' values were significantly reduced in all ROIs of PD and PSP patients, compared with controls (p=0.001). V3'' value in ROI 2 was significantly lower in PSP group, compared with PD group (p=0.02). The percent reductions of striatal uptake in ROI 1, ROI 2, ROI 3 and ROI 4 were 56%, 53%, 64% and 78% in PD patients, whereas 75%, 72%, 75% and 77% in PSP patients, respectively. The reduction patterns of uptake were significantly different between PD and PSP groups (p=0.001). In PD patients, the percent reductions of (123)I-IPT uptake were significantly greater in ROI 3 and 4 compared with ROI 1 or 2, whereas those were similar in all ROIs of PSP patients. In addition, PD patients showed a significantly higher posterior putamen/caudate ratio of reduced (123)I-IPT uptake than the anterior putamen/caudate ratio (p=0.005). Our results implicate that (123)I-IPT SPECT is a relatively simple and reliable technique that may be useful in differentiating PD from PSP. PMID- 16473372 TI - Notch-1 immunoexpression is increased in Alzheimer's and Pick's disease. AB - Notch-1 is a protein that influences cell fate decisions, with its expression occurring primarily during embryogenesis and development. However, Notch-1 is also expressed in the adult brain, in regions with high synaptic plasticity, particularly the hippocampus. Its role in adults is unknown; however, it may impact neurite outgrowth or cell differentiation in adult brain regions undergoing neurogenesis. Notch-1 is increased in Alzheimer's disease (AD); however, its expression in other CNS degenerative diseases has not been described. To begin to define the range of degenerative disorders where Notch-1 expression is altered, we examined Notch-1 immunoreactivity in a variety of neurodegenerative diseases to determine whether its increase is selective for AD. We examined sections of hippocampus from 13 AD, 13 classical Pick's disease (PiD; with Pick bodies), 4 dementia lacking distinctive histopathology (DLDH) and 8 control brains, emphasizing hippocampal (dentate gyrus) pathology. We determined that Notch-1 immunoexpression is increased in AD and PiD relative to control cases. DLDH cases were not significantly different than control cases with respect to Notch-1 expression. Given the increase in Notch-1 immunoexpression in AD and PiD, two diseases where abnormal tau aggregates are present, and the lack of Notch-1 immunoexpression in DLDH (where tau aggregates are absent), we cannot rule out the possibility that tau aggregates are associated with Notch-1 expression in neurodegenerative diseases. PMID- 16473373 TI - A simple time delay model for eukaryotic cell cycle. AB - We propose a seven variable model with time delay in one of the variables for the cell cycle in higher eukaryotes. The model consists of four important phosphorylation-dephosphorylation (P-D) cycles that govern the cell cycle, namely Pre-MPF-MPF, Cdc25P-Cdc25, Wee1P-Wee1 and APCP-APC. Other variables are cyclin, free cyclin dependent kinase (Cdk) and mass. The mass acts as a G2/M checkpoint and the checkpoint is represented by a saddle node loop bifurcation. The key feature of the model is that a time lag has been introduced in the activation of anaphase promoting complex (APC) by maturation promoting factor (MPF). This is effected by treating MPF as a time-delayed variable in the activation step of APC. The time lag acts as a spindle checkpoint. Absence of time delay induces a bistability in our model. Time delay also brings about variability in G1 phase timings. The model also reproduces the mutant phenotype experiments on wee1 cells. Stochasticity has been introduced in the model to simulate the dependence of the cycle time on cell birth length. Mutant phenotypes in the stochastic model reproduce the experimental observations better than the deterministic model. PMID- 16473374 TI - Inhibition of inducible nitric oxide synthesis by azathioprine in a macrophage cell line. AB - Azathioprine is used as an anti-inflammatory agent. Although there are numerous data demonstrating cytotoxic and immunosuppressive properties of azathioprine and its metabolite 6-mercaptopurine, the mechanism of the anti-inflammatory action of azathioprine has not yet been fully clarified. During our study, we investigated the effects of azathioprine on the inducible nitric oxide synthase (iNOS) in lipopolysaccharide stimulated murine macrophages (RAW 264.7) by measurement of iNOS protein (immunoblotting), iNOS mRNA (semiquantitative competitive RT-PCR), and NO production (nitrite levels). Azathioprine (0-210 muM) induces a concentration dependent inhibition of inducible nitric oxide synthesis (IC50: 33.5 muM). iNOS protein expression showed a concentration dependent reduction as revealed by immunoblotting when cells were incubated with increasing amounts of azathioprine. Azathioprine decreases iNOS mRNA levels as shown by semiquantitative competitive RT-PCR. In contrast, 6-mercaptopurine showed no inhibition of inducible nitric oxide synthesis. Azathioprine did not reduce iNOS mRNA stability after the addition of actinomycin D. Enzymatic activity assays with increasing concentrations of azathioprine (0-210 muM) showed no statistically significant inhibition of iNOS enzyme activity compared to cell lysates without azathioprine. Nuclear translocation of NF-kappaB p65 subunit and binding of NF-kappaB p50 subunit from nuclear extracts to a biotinylated consensus sequence was unaffected by azathioprine treatment. iNOS inhibition by azathioprine was associated with a decreased expression of IRF-1 (interferon regulatory factor 1) and IFN-beta (beta-interferon) mRNA. Azathioprine induced iNOS inhibition seems to be associated with an action of the methylnitroimidazolyl substituent. This suggests a route to the rational design of nontoxic anti-inflammatory agents by replacing the 6-mercaptopurine component of azathioprine with other substituents. The inhibition of inducible nitric oxide synthesis might contribute to the anti-inflammatory activities of azathioprine. PMID- 16473375 TI - The equity of access to orthodontic dental care for children in the North East of England. AB - OBJECTIVES: To assess the equity of access to primary care orthodontic treatment in relation to deprivation in the County Durham and Tees Valley Strategic Health Authority area. STUDY DESIGN: An observational study based on Dental Practice Board data for the County Durham and Tees Valley Strategic Health Authority area with a population of 1.13 million. METHODS: The postcode of all orthodontic claims made by National Health Service dentists across the area in the financial years 2002/2003 and 2003/2004 were obtained and the claim rate per 1000 at risk population calculated for each ward. These ward rates were then compared to both the level of deprivation measured by the Index of Multiple Deprivation 2000 and the children's dental registration rate of the area. RESULTS: Inequity in access to orthodontic care was observed. There was a moderate negative correlation r = 0.40 suggesting wards with the lowest claim rates had the greatest deprivation. In addition, the wards with the lowest child dental registration rates also had the lowest claim rates for orthodontic treatment. CONCLUSIONS: Currently, there is inequity of access to orthodontic treatment for children in County Durham and Tees Valley. The move towards local commissioning for dental services within the NHS will provide an opportunity to reduce inequalities in access. PMID- 16473376 TI - Factors associated with non-participation in a physical activity promotion trial. AB - BACKGROUND: Non-participation can bias outcome in intervention studies of physical activity. OBJECTIVES: To compare characteristics, knowledge and attitudes to physical activity in participants and non-participants of a physical activity intervention trial in primary care. STUDY DESIGN: Cross-sectional survey. METHODS: Patients aged 40-64 years were recruited opportunistically during surgery visits in an inner city general practice in Newcastle upon Tyne, UK. Attitudes to physical activity, views of its health benefits, and barriers to participation were elicited in interviews with participants, and by postal questionnaire from non-participants. Data held by general practitioners were used to compare anthropometry and lifestyle between groups. RESULTS: Of 842 eligible patients, 276 (33%) refused outright (non-volunteers) and 566 volunteered for the intervention study, of which 353 (42%) attended a baseline assessment and 213 (25%) subsequently defaulted. The initial refusal rate was higher amongst men, smokers and those with addresses in more deprived areas. The response rate to the postal survey of non-volunteers was 45%. Compared with participants, the non volunteers were more likely to be an adult carer and to report poorer health, and were less likely to have had higher education or to have children living at home. Far more non-volunteers considered that they already did enough exercise to maintain health. Non-volunteers had slightly less knowledge of the benefits of physical activity; attached far less importance to it in maintaining health; were more likely to cite 'fear of leaving their home unattended', 'do not enjoy exercise' and 'poor health' as barriers to exercise; and were less likely to cite 'no one to exercise with' as a barrier to exercise. CONCLUSION: Recruitment of 'hard to engage' individuals requires careful phrasing of the message to focus on their personal goals and to address gaps in their knowledge about physical activity and the principal barriers they perceive. Differential uptake across population subgroups could lead to a widening of health inequalities. PMID- 16473377 TI - HIV-related sexual behaviour among migrants and non-migrants in a rural area of China: role of rural-to-urban migration. AB - OBJECTIVE: To compare human immunodeficiency virus (HIV)-related sexual risks between temporary rural-to-urban migrants and non-migrants, and to explore the role of migration in HIV transmission in a rural area of China. STUDY DESIGN: A cross-sectional study was conducted among 605 (302.5 couples, one male missing) marriage licence applicants. METHODS: A two-part anonymous questionnaire was used. The first part included non-sensitive questions including demographics and HIV knowledge. Sensitive questions related to sexual behaviours were administered in the second part of the questionnaire using a tape recorder, earphones and an answer sheet. RESULTS: Sixty-six percent (399/605) of the study subjects had migrated to a city for a temporary job. The proportions of migrants who had premarital sex (62%, 247/399) and multiple sexual partners (12%, 47/399) were significantly higher than those of non-migrants (52 and 6%, respectively). Among those who had multiple sexual partners, only 9% (4/47) of migrants and 8% (1/12) of non-migrants reported often or always using condoms with sexual partners other than their spouse. A low level of knowledge regarding HIV and acquired immunodeficiency syndrome was observed in both groups. Very few migrants (15%, 61/399) and non-migrants (10%, 20/206) perceived themselves to be at risk of HIV infection. CONCLUSIONS: As both migrants and non-migrants are at risk of HIV infection, intervention programmes targeting reduction of HIV sexual risk behaviour should cover both groups in rural areas of China. PMID- 16473378 TI - Access to water in the countries of the former Soviet Union. AB - BACKGROUND: During the Soviet period, authorities in the USSR invested heavily in collective farming and modernization of living conditions in rural areas. However, many problems remained, including poor access to many basic amenities such as water. Since then, the situation is likely to have changed; economic decline has coincided with migration and widening social inequalities, potentially increasing disparities within and between countries. AIM: To examine access to water and sanitation and its determinants in urban and rural areas of eight former Soviet countries. METHODS: A series of nationally representative surveys in Armenia, Belarus, Georgia, Moldova, Kazakhstan, Kyrgyzstan, Russia and Ukraine was undertaken in 2001, covering 18,428 individuals (aged 18+ years). RESULTS: The percentage of respondents living in rural areas varied between 27 and 59% among countries. There are wide urban-rural differences in access to amenities. Even in urban areas, only about 90% of respondents had access to cold running water in their home (60% in Kyrgyzstan). In rural areas, less than one third had cold running water in their homes (44% in Russia, under 10% in Kyrgyzstan and Moldova). Between one-third and one-half of rural respondents in some countries (such as Belarus, Kazakhstan and Moldova) obtained their water from wells and similar sources. Access to hot running water inside the homes was an exception in rural households, reflecting the lack of modern heating methods in villages. Similarly, indoor access to toilets is common in urban areas but rare in rural areas. Access to all amenities was better in Russia compared with elsewhere in the region. Indoor access to cold water was significantly more common among rural residents living in apartments, and in settlements served by asphalt roads rather than dirt roads. People with more assets or income and living with other people were significantly more likely to have water on tap. In addition, people who had moved in more recently were more likely to have an indoor water supply. CONCLUSIONS: This was the largest single study of its kind undertaken in this region, and demonstrates that a significant number of people living in rural parts of the former Soviet Union do not have indoor access to running water and sanitation. There are significant variations among countries, with the worse situation in central Asia and the Caucasus, and the best situation in Russia. Access to water strongly correlates with socio-economic characteristics. These findings suggest a need for sustained investment in rebuilding basic infrastructure in the region, and monitoring the impact of living conditions on health. PMID- 16473379 TI - A randomized controlled trial to assess the effectiveness of a letter from a consultant surgeon in causing smokers to stop smoking pre-operatively. PMID- 16473380 TI - Prismatic dentine in the Australian lungfish, Neoceratodus forsteri (Osteichthyes: Dipnoi). AB - The Australian lungfish, Neoceratodus forsteri, has a dentition consisting of enamel, mantle dentine and bone, enclosing circumdenteonal, core and interdenteonal dentines. Branching processes from cells that produce interdenteonal dentine leave the cell surface at different angles, with collagen fibrils aligned parallel to the long axis of each process. In the interdenteonal dentine, crystals of calcium hydroxyapatite form within fibrils of collagen, and grow within a matrix of non-collagenous protein. Crystals are aligned parallel to the cell process, as are the original collagen fibrils. Because the processes are angled to the cell surface, the crystals within the core or interdenteonal dentine are arranged in bundles set at angles to each other. Apatite crystals in circumdenteonal dentine are finer and denser than those of the interdenteonal dentine, and form outside the fibrils of collagen. In mature circumdenteonal dentine the crystals of circumdenteonal dentine form a dense tangled mass, linked to interdenteonal dentine by isolated crystals. The functional lungfish tooth plate contains prisms of large apatite crystals in the interdenteonal dentine and masses of fine tangled crystals around each denteon. This confers mechanical strength on a structure with little enamel that is subjected to heavy wear. PMID- 16473381 TI - Prenatal cigarette smoke exposure: pregnancy outcome and gestational changes in plasma nicotine concentration, hematocrit, and carboxyhemoglobin in a newly standardized rat model. AB - Epidemiological studies support an association between perinatal cigarette smoke (CS) exposure and a number of severe pre- and postnatal complications. However, the mechanisms through which CS enhances such risks largely remain unknown. One of the reasons for our inability to discover such mechanisms has been the unavailability of a clinically relevant and physiologically concordant animal model. A number of studies have previously used nicotine (Nic) as surrogate for CS. We sought to (1) establish the amount of CS exposure to achieve plasma Nic concentrations observed among moderate to heavy smokers (20-60 ng/ml), (2) investigate the temporal changes in plasma Nic concentrations, carboxyhemoglobin, and hematocrit with advancing pregnancy, and (3) elucidate the effects of CS exposure on pregnancy outcome. Pregnant Sprague-Dawley rats were exposed to various doses of CS or room air (Sham) from days 6 to 21 of gestation. Exposure to 6000 ml/day of CS led to very high plasma Nic concentrations and increased maternal and fetal mortality (P < 0.001). The plasma Nic concentrations remained higher than those observed in moderate smokers until the CS dose was reduced to 1000 ml/day and showed dose-dependent temporal changes with advancing gestational age. Significant increases in carboxyhemoglobin and hematocrit were observed in the CS group as compared with the Sham group (P < 0.001). In addition, prenatally CS exposed fetuses had lower birth weight as compared with the Sham group (P = 0.04). Our current study establishes a newly standardized and physiologically relevant model to investigate the mechanisms of CS-mediated adverse effects during the critical period of fetal development. PMID- 16473382 TI - Oxidative stress-driven mechanisms of nordihydroguaiaretic acid-induced apoptosis in FL5.12 cells. AB - Nordihydroguaiaretic acid (NDGA), a general lipoxygenase (LOX) enzyme inhibitor, induces apoptosis independently of its activity as a LOX inhibitor in murine pro B lymphocytes (FL.12 cells) by a mechanism that is still not fully understood. Glutathione depletion, oxidative processes and mitochondrial depolarization appear to contribute to the apoptosis induced by NDGA. The current data demonstrate that NDGA (20 microM)-induced apoptosis in FL5.12 cells is partially protected by N-acetylcysteine (NAC) (10 mM) and dithiothreitol (DTT) (500 microM) pretreatment, confirming a role for oxidative processes. In addition, the treatment of FL5.12 cells with NDGA led to an increase in phosphorylation and activation of the MAP kinases ERK, JNK and p38. Although pretreatment with ERK inhibitors (PD98059 or U0126) abolished ERK phosphorylation in response to NDGA, neither inhibitor had any effect on NDGA-induced apoptosis. SP600125, a JNK inhibitor, did not have any effect on NDGA-induced phosphorylation of JNK nor apoptosis. Pretreatment with the p38 inhibitor SB202190 attenuated NDGA-induced apoptosis by 30% and also abolished p38 phosphorylation, compared to NDGA treatment alone. NAC, but not DTT, also decreased the phosphorylation of p38 and JNK supporting a role for oxidative processes in activating these kinases. Neither NAC nor DTT blocked the phosphorylation of ERK suggesting that this activation is not related to oxidative stress. The release of cytochrome c and activation of caspase-3 induced by NDGA were inhibited by NAC. SB202190 slightly attenuated caspase-3 activation and had no effect on the release of cytochrome c. These data suggest that several independent mechanisms, including oxidative reactions, activation of p38 kinase and cytochrome c release contribute to NDGA induced apoptosis. PMID- 16473383 TI - The effects of dexamethasone on rat brain cortical nuclear factor kappa B (NF kappaB) in endotoxic shock. AB - To explore the molecular mechanism of brain tissue injury induced by lipopolysaccharide (LPS), we studied the effects of endotoxic shock on rat brain cortex NF-kappaB and the effects of dexamethasone on these changes. Rats were randomly divided into LPS, LPS + dexamethasone, and control groups. The DNA binding activity of NF-kappaB was observed using electrophoretic mobility shift assay (EMSA). Protein expression in nuclear extracts was studied using Western blots, and nuclear translocation was observed using immunohistochemistry. These indices were assayed at 1 h and 4 h after intravenous injection of LPS (4 mg x kg(-1)). EMSA showed significantly increased NF-kappaB DNA-binding activity in nuclear extracts from the LPS group at both 1 h and 4 h after LPS injection, compared with the control group (P < 0.01). For the LPS group, the NF-kappaB DNA binding activity was greater at 1 h than at 4 h (P < 0.05). The expression of p65 and p50 protein in the nuclear extracts was also increased, as compared with the control group. However, the expression of p65 and p50 protein from cytosolic extracts did not show any significant change. Dexamethasone down-regulated not only NF-kappaB DNA-binding activity but also the expression of p65 protein in the nuclear extracts. From these data, we have concluded that NF-kappaB activation and nuclear translocation of NF-kappaB play a key role in the molecular mechanism of brain tissue injury in endotoxic shock. Dexamethasone may alleviate brain injury by inhibiting NF-kappaB activation. PMID- 16473384 TI - Combining mutations in HIV-1 reverse transcriptase with mutations in the HIV-1 polypurine tract affects RNase H cleavages involved in PPT utilization. AB - The RNase H cleavages that generate and remove the polypurine tract (PPT) primer during retroviral reverse transcription must be specific to generate linear viral DNAs that are suitable substrates for the viral integrase. To determine if specific contacts between reverse transcriptase (RT) and the PPT are a critical factor in determining the cleavage specificity of RNase H, we made HIV-1 viruses containing mutations in RT and the PPT at the locations of critical contacts between the protein and the nucleic acid. The effects on titer and RNase H cleavage suggest that combining mutations in RT with mutations in the PPT affect the structure of the protein of the RT/nucleic acid complex in ways that affect the specificity and the rate of PPT cleavage. In contrast, the mutations in the PPT (alone) and RT (alone) affect the specificity of PPT cleavage but have much less effect on the overall rate of cleavage. PMID- 16473385 TI - The parallel programming of voluntary and reflexive saccades. AB - A novel two-step paradigm was used to investigate the parallel programming of consecutive, stimulus-elicited ('reflexive') and endogenous ('voluntary') saccades. The mean latency of voluntary saccades, made following the first reflexive saccades in two-step conditions, was significantly reduced compared to that of voluntary saccades made in the single-step control trials. The latency of the first reflexive saccades was modulated by the requirement to make a second saccade: first saccade latency increased when a second voluntary saccade was required in the opposite direction to the first saccade, and decreased when a second saccade was required in the same direction as the first reflexive saccade. A second experiment confirmed the basic effect and also showed that a second reflexive saccade may be programmed in parallel with a first voluntary saccade. The results support the view that voluntary and reflexive saccades can be programmed in parallel on a common motor map. PMID- 16473386 TI - Collinear context (and learning) change the profile of the perceptual filter. AB - The effect of collinear context on the filter mediating the detection of a Gabor stimulus was investigated by using the classification image method. Classification images were estimated for a 1.5 cpd horizontal Gabor target and the same target flanked by two collinear Gabors horizontally 1.7 degrees displaced from the target. The target was masked by a low-contrast white-noise mask. Obtained classification images were fitted by Gabor functions. The results show that collinear flankers increase the length of the classification image profiles along the collinear axis. At the same time, modest facilitory effects were observed in most subjects. The specificity and the amount of context-induced elongation in the classification images makes it hard to be explained by uncertainty reduction alone. In previous studies, collinear facilitation has been reported to abolish due to perceptual learning. We report a possibly related phenomenon: classification image data was re-analyzed in two parts consisting of the early and the late trials. In the latter trials, differences between the classification images in flankers and no-flankers condition are no longer significant. PMID- 16473387 TI - Late stages of visual pigment photolysis in situ: cones vs. rods. AB - Slow photolysis reactions and the regeneration of the dark pigment constitute the mechanisms of dark adaptation whereby photoreceptor cells restore their sensitivity after bright illumination. We present data on the kinetics of the late stages of the photolysis of the visual pigment in intact rods and red- and green-sensitive cones of the goldfish retina. Measurements were made on single photoreceptors by means of a fast-scanning dichroic microspectrophotometer. We show that in cones the hydrolysis of the opsin-all-trans 3-dehydroretinal linkage proceeds with a half-time of approximately 5s at 20 degrees C that is almost two orders of magnitude faster than in rods. 3-Dehydroretinol in cones is produced approximately 3-fold faster than retinol in amphibian rhodopsin rods; the rate of the reaction is limited by the speed of retinal reduction catalyzed by retinoldehydrogenase. The fast hydrolysis of the 3-dehydroretinal/opsin Schiff base and the correspondingly fast appearance of the substrates for dark visual pigment regeneration (free opsin and 3-dehydroretinol) provide essential conditions for faster dark adaptation of cone (diurnal) as compared to rod (nocturnal) vision. PMID- 16473388 TI - Comment on "Chromate ion adsorption by agricultural by-products modified with dimethyloldihydroxylethylene urea and choline chloride" by Wartelle and Marshall. PMID- 16473389 TI - Biotransformation of the flame retardant tetrabromo-bisphenol A by human and rat sub-cellular liver fractions. AB - The comparative in vitro metabolism of the flame retardant tetrabromo-bisphenol A was studied in rat and human using a [(14)C]-radio-labelled molecule. Tetrabromo bisphenol A is metabolised into the corresponding glucuronide (liver S9 fractions) and several other metabolites produced by cytochrome P450 dependent pathways (liver microsomes and liver S9 fractions). No major qualitative differences were observed between rat and human, regardless of the selected concentration, within the 20-200 microM range. Tetrabromo-bisphenol A undergoes an oxidative cleavage near the central carbon of the molecule, that leads to the production of hydroxylated dibromo-phenol, hydroxylated dibromo-isopropyl-phenol and glutathione conjugated dibromo-isopropyl-phenol. The main metabolites of tetrabromo-bisphenol A are two molecules of lower polarity than the parent compound, characterised as a hexa-brominated compound with three aromatic rings and a hepta-brominated dimer-like compound, respectively. Both structures, as well as the lower molecular weight metabolites resulting from the breakdown of the molecule, suggest the occurrence of chemically reactive intermediates formed following a first step oxidation of tetrabromo-bisphenol A. PMID- 16473390 TI - Sorption-desorption behavior of PCP on soil organic matter and clay minerals. AB - Pentachlorophenol (PCP) contamination is a severe environmental problem due to its widespread occurrence, toxicity and recalcitrance. In order to gain a better understanding of the fate of PCP in soils, the role of the soil organic matter (SOM) and clay minerals in the PCP sorption-desorption was studied on two bulk field soils, two subsoils (i.e., SOM or clay-removed soil) and two artificial soils. The two field soils used were a silty loam from New Mexico (NM) containing 10% clay and a sandy-clay-loam from Colombia (CO) South America comprised of 18% clay minerals. The bulk CO soil containing kaolinite sorbed significantly less PCP than the NM soil. All soils depicted an apparent hysteresis during sorption. The CO bulk and subsoils desorbed 14-20% and 15-26% of the sorbed PCP respectively whereas the NM bulk and subsoils desorbed only 4-12% and 5-16%, respectively. Experiments conducted with pure clay and artificial soils indicated that the expandable clay minerals were key sorbent material. Additional studies to investigate the interaction between SOM and clay minerals are needed to fully understand sorptive phenomena. PMID- 16473391 TI - Photochemically-assisted electrochemical degradation of landfill leachate. AB - In this work, the treatment of landfill leachates by a photoelectrochemical procedure is reported. When applied to untreated leachates the photochemical system was significantly hindered on account of the characteristic dark coloration of the samples. At this condition the degradation process was essentially electrochemical permitting typical color and COD removal of about 50% and 20%, respectively. When a previous chemical precipitation process was applied aiming the elimination of colored species (mainly humic substances) the decolorization and COD removal was extended to 90% and 60%, respectively. Considering the extremely complex character of the leachates and its usual resistance to conventional degradation processes the result reported here attest the high potentiality of photoelectrochemical processes to remediation of recalcitrant residues. PMID- 16473392 TI - Identification and steroid receptor activity of products formed from the bromination of technical nonylphenol. AB - Alkylphenols are commonly present in wastewater effluents and may contribute to the total hormonal loading of receiving waters due to their weakly estrogenic properties. However the presence of reactive bromine species in some treated wastewaters can result in the formation of brominated alkylphenols which may also possess steroid receptor activity. In this study, the products of bromination of technical nonylphenol (NP) were identified, purified and tested in vitro in recombinant yeast steroid receptor transcription assays. Bromination of NP in the presence of acetic acid resulted in the formation of one major product which was identified as 2-bromo-4-nonylphenol (NPBr). In the presence of methanol/water, bromination of NP resulted in the formation 2,6-dibromo-4-nonylphenol (NPBr2) as well as a number of other minor polybrominated products. The EC50 of NPBr in the yeast estrogen receptor transcription (YES) assay was 6.7x10(-6) M, which was 48 fold less active than NP and 86,000 fold less active than the estrogen agonist 17beta-estradiol NPBr2 was not active in the YES assay. NP, NPBr and NPBr2 were all weakly androgenic in the yeast androgen receptor transcription assay but at concentrations which were 100,000 fold less active than the androgen receptor agonist dihydrotestosterone. Neither NP, NPBr or NPBr2 exhibited appreciable anti estrogenic or anti-androgenic activity in the yeast receptor transcription assays. This study suggests that bromination of NP markedly reduces its estrogen receptor transcription activity but has no effect on the weak androgen receptor transcription activity of the alkylphenol. PMID- 16473393 TI - Coupling of 2,4,6-trinitrotoluene (TNT) metabolites onto humic monomers by a new laccase from Trametes modesta. AB - During degradation of trinitrotoluene (TNT) by Trametes modesta, addition of humic monomers prevented the accumulation of all major stable TNT metabolites (aminodinitrotoluenes [AMDNT]) by at least 92% in the presence of 200 mM ferulic acid and guaiacol. Acute toxicity tests with individual TNT metabolites and in T. modesta cultures supplemented with 200 microM TNT demonstrated that the TNT biodegradation process lead to less toxic metabolites. Toxicity decreased in the order TNT>4-HADNT (4-hydroxylaminodinitrotoluene)>2-HADNT>2,6-DNT (2,6 dinitrotoluene)>2',2',6,6-azoxytetranitrotoluene>4-AMDNT>2-AMDNT>2,4 diamninonitrotoluene (2,4-DAMNT) while 2,4-DNT and 2,6-DAMNT were the least toxic. Ferulic acid is the best candidate for immobilization TNT biodegradation metabolites since it prevented the accumulation of AMDNTs in cultures during TNT biodegradation and its products were less toxic. All humic monomers were very effective in immobilizing 2-HADNT [100%], 4-HADNT [100%] and 2,2,6,6 azoxytetranitrotoluene [100%]. Two distinct laccase isoenzymes (LTM1 and LTM2) potentially involved in immobilization of TNT degradation products were purified to electrophoretic homogeneity. LTM1 and LTM2 have molecular weights of 77.6 and 52.5 kDa, are 18% and 24% glycosylated, have pI values of 3.6 and 4.2, respectively. Both enzymes oxidized all the typical laccase substrates tested. LTM1 showed highest kinetic constants (K(m)=0.03 microM; K(cat)=8.8 4x 10(7)s( 1)) with syringaldazine as substrate. PMID- 16473394 TI - The application of electrochemical technology to the remediation of oily wastewater. AB - The successful application of electrochemical technology, employing a dimensionally stable anode (DSA((R))), for the remediation of wastewater from the oil extraction industry has been demonstrated. Samples from the oil-water separation box of an effluent treatment plant were submitted to voltammetry, chronoamperometry and electrolysis studies using a DSA anode of nominal composition Ti/Ru(0.34)Ti(0.66)O(2). Electrolysis of the oily wastewater lead to a time-dependent reduction in chemical oxygen demand (COD) in the sample that could be attributed to: (i) the direct oxidation of oil components at the electrode, by the metal oxide itself or by OH() radicals available at the electrode surface, (ii) the indirect oxidation of oil components by intermediate oxidising agents formed in parallel reactions (ex. ClO(-)), and (iii) the aggregation of suspended oil droplets by electroflotation. The largest reduction (57%) in COD was obtained following electrolysis of an oily sample for 70 h at 50 degrees C with a current density of 100 mA cm(-2). The stability of DSA electrodes for use in oily wastewater remediation has been assessed. PMID- 16473395 TI - Remazol Brilliant Blue R decolourization by the laccase from Trametes trogii. AB - The decolourization of the recalcitrant dye RBBR by the culture filtrate of Trametes trogii and its isolated laccase was investigated. Both filtrates from Cu induced cultures as well as purified laccase decolourized the dye RBBR. The purified laccase decolourized the dye down to 97% of 100 mg l(-1) initial concentration of RBBR when only 0.2U ml(-1) of laccase was used in the reaction mixture. The effects of different physicochemical parameters were tested and optimal decolourization rates occurred at pH 5 and at a temperature of 50 degrees C. Decolourization of RBBR occurred in the presence of metal ions which could be found in textile industry effluents. Of all the metal ions tested, FeCl2 was the most inhibiting for the decolourization. HBT was shown to have no effect on the decolourization of RBBR at low concentration, while at a concentration of 5 mM it slightly inhibited decolourization. The presence of aromatic compounds was found to be inhibiting for the decolourization at a concentration of 10 mM, but not at 0.1 mM, while at 1 mM only ortho-diphenols were inhibiting. Probing the effect of methanol it was found that higher concentrations caused a decrease in the decolourization rate of RBBR. The effect of laccase inhibitors on the decolourization of RBBR was tested with L-cysteine, SDS and EDTA. It was demonstrated that L-cysteine was the most inhibiting substrate for the decolourization while SDS was only inhibiting at 10 mM concentration and ETDA was not inhibiting at all tested concentrations. PMID- 16473396 TI - Acetylsalicylic acid is compounding to antiplatelet effect of C-reactive protein. AB - The contribution of inflammatory process to the modulation of platelet response to acetylsalicylic acid (ASA) remains obscure. In our study, we examined the in vitro effect of C-reactive protein (CRP) on the ASA-mediated inhibition of collagen-stimulated platelet reactivity. Influence of CRP on platelet responsiveness to ASA was analysed using classical turbidimetric aggregation and flow cytometry. When acting alone, both C-reactive protein and ASA inhibited collagen-dependent platelet aggregation and reduced the expressions of two platelet surface membrane activation markers: P-selectin and activated GPIIbIIIa complex. Compared to the effects observed for ASA alone, the simultaneous action of both agents lead to further reductions in platelet aggregation (by 56.7+/-1.0% vs. 14.9+/-0.6%, p<0.0001) and lowered the expressions of platelet surface membrane P-selectin (by 72.1+/-5.3% vs. 65.0+/-6.0%, p<0.01) and activated GPIIbIIIa (by 67.0+/-5.6% vs. 47.7+/-8.3%, p<0.01). In general, our findings showed for the first time the augmenting effect of native C-reactive protein in the antiplatelet action of acetylsalicylic acid. Thus, we conclude that the effectiveness of aspirin therapy may strongly depend upon the presence of native CRP in circulation. PMID- 16473397 TI - Role of prophylactic hysterectomy in patients at high risk for hereditary cancers. AB - BACKGROUND: Current surgical recommendations for ovarian cancer prophylaxis in women at high risk of developing ovarian cancer include bilateral salpingo oophorectomy (risk-reducing salpingo-oophorectomy (RRSO)). The role of hysterectomy is unclear. We sought to determine outcomes following prophylactic surgery in high-risk women. METHODS: We surveyed unaffected members of the Gilda Radner Familial Ovarian Cancer Registry who had undergone oophorectomy from 1981 to 2002. Data were collected and analyzed for statistical significance by the Fisher's Exact Test. RESULTS: Two hundred eighty women were surveyed, and 154 (55%) responded; 97% were Caucasian and 14% reported being Jewish. The median age of the respondents was 51 years (range 29-79); median age at oophorectomy was 41 years (range 15-68). Fifty-eight patients (38%) reported a laparoscopic procedure. One hundred five patients (68%) had a simultaneous hysterectomy, and 4 (3%) had a prior hysterectomy. Forty-four patients (29%) underwent BSO only. Of these 44 patients, 40 (91%) did not require a subsequent hysterectomy. Of the 4 who did, 2 were for leiomyomas, one for menorrhagia and the other was unknown. While not statistically significant, of the 3 patients who developed a subsequent gynecologic malignancy, all had undergone a hysterectomy. There was a statistically significant difference in whether or not the uterus was removed as part of the procedure by time period, whereby women treated prior to 1990 had a higher likelihood of having a hysterectomy (P = 0.03). CONCLUSION: The women in our study did not require hysterectomy for prevention of malignancy. We conclude that one should screen for benign gynecological indications for hysterectomy when planning a prophylactic BSO for prevention of ovarian cancer. Other potential risk factors for endometrial cancer, including the role of UPSC in HBOC, remain to be elucidated. PMID- 16473398 TI - Association between endometriosis and cancer: a comprehensive review and a critical analysis of clinical and epidemiological evidence. AB - OBJECTIVE: This review was aimed to critically evaluate observational, cohort, and case-control studies performed so far in order to assess the association between endometriosis and malignant diseases. Based on the observations herein presented, clinical indications that might avoid physicians' mismanaging of affected patients are proposed. METHODS: Search strategies included online searching of the MEDLINE database and hand searching of relevant publications and reviews. Additional reports were collected by systematically reviewing all references from retrieved papers. RESULTS: Endometriosis is not associated with an increased risk of cancer in general. Data from large cohort and case-control studies indicate an increased risk of ovarian cancers in women with endometriosis. The observed effect sizes are modest varying between 1.3 and 1.9. Evidence from clinical series consistently demonstrates that the association is confined to the endometrioid/clear-cell histotypes. Available studies are characterized by several limitations, some of which potentially bias results towards the null hypothesis whereas others leading to overestimate the association. Evidence for an association with melanoma and non-Hodgkin's lymphoma is increasing but still to be verified whereas an increased risk for other gynecological cancer types is not supported. CONCLUSIONS: Epidemiological findings on the association between endometriosis and cancer are still elusive. At present, endometriosis should not be considered a medical condition associated with a clinically relevant risk of any specific cancer. On the basis of the present findings, modifications of the standard treatment options for the disease are not justifiable. PMID- 16473399 TI - A case of a pT3, HPV 52-positive vulvar carcinoma in an 18-year-old woman. AB - BACKGROUND: Vulvar carcinoma in young women is rare; so far, no case of an 18 year-old woman has been described. Here, we report a case with a T3 HPV 52 induced tumor 3 years after primary HPV contamination. CASE: An 18-year-old woman without risk factors complaining of dysuria and vulvar pain was treated several months for fungal infection before referred and diagnosed with a vulvar carcinoma located between clitoris and urethra. She underwent operation with partial urethral resection and external radiation. The tumor tested positive for HPV type 52, the time between primary sexual contact and tumor development was less than 3 years. CONCLUSION: Also in very young women, an ulcer and vulvar pain have to be biopsied to exclude malignancy despite an unusual short time interval between possible HPV contamination and symptoms. PMID- 16473400 TI - Degeneration and apoptosis of endometrial cells in the bitch. AB - The relationships between changes in plasma progesterone concentrations, degeneration of the luminal epithelium, the occurrence of apoptosis of endometrial cells and endometrial leucocyte populations in the bitch were determined. Mature bitches (n = 15) were euthanized and necropsied when in diestrus (Days 7-75, n = 12) or in anestrus (Days 10, 32 and 53). Degeneration of the luminal epithelium was observed in bitches in late diestrus (Days 38-75, n = 5) when plasma progesterone concentrations were decreasing and in anestrus (Days 10 and 32, n = 2) when plasma progesterone concentrations were < 0.5 ng/mL. Endometrial leucocyte populations increased after degeneration of the luminal epithelium (around Day 42 of diestrus). Apoptosis was mainly observed in the basal glandular epithelial cells and endothelial cells of blood capillaries in all except anestrous bitches. Very few apoptotic cells were found in the superficial glandular epithelial cells and stromal cells. Higher apoptotic indices were detected in the basal glandular epithelium on Days 12-42 of diestrus than at other stages. Therefore, apoptosis of glandular basal epithelial cells occurred mainly in early diestrus, degeneration of cells of the luminal epithelium occurred from mid-diestrus to early anestrus, and the increase in leucocyte numbers may have been a consequence and not a cause of luminal epithelial degeneration. PMID- 16473401 TI - In vitro development of domestic cat embryos following intra-cytoplasmic sperm injection with testicular spermatozoa. AB - The objective was to assess the ability of testicular spermatozoa to fertilize in vitro matured domestic cat oocytes and support blastocyst formation in vitro following intra-cytoplasmic sperm injection (ICSI). After IVM, oocytes were randomly and equally allocated among treatment groups (ICSI with testicular spermatozoa, ICSI with ejaculated spermatozoa, sham ICSI, and control IVF). At 18 h after either injection or insemination, the percentage of fertilized oocytes (per total metaphase II oocytes) was approximately 65% after ICSI with testicular or ejaculated spermatozoa (P > 0.05), which was less (P < 0.05) than control IVF (approximately 90%). On Day 7, the percentage of cleaved embryos (per total metaphase II oocytes) was approximately 60% after ICSI with testicular or ejaculated spermatozoa (P > 0.05), which also was less (P < 0.05) than control IVF (approximately 85%). After ICSI with testicular spermatozoa, the percentage of blastocysts (per total cleaved embryos) was approximately 11.0%, which was less (P < 0.05) than ICSI with ejaculated spermatozoa (approximately 21.0%); the latter was less (P < 0.05) than control IVF (approximately 43.0%). No blastocyst formation was observed after sham ICSI. For the first time in the domestic cat, this study demonstrated the fertilizing ability and developmental potential of intra-testicular spermatozoa delivered directly into intra-ovarian oocytes matured in vitro. PMID- 16473402 TI - Fatigue of zirconia under cyclic loading in water and its implications for the design of dental bridges. AB - OBJECTIVES: The aim of this study was to evaluate the cyclic fatigue behavior of zirconia (3Y-TZP) in water and derive guidelines for the design of zirconia-based dental bridges with extended lifetime. METHODS: The subcritical crack growth parameters under aqueous and cyclic loading conditions were determined from Weibull distributions of the initial mechanical strength and of the lifetime of TZP specimens. The strength and lifetime data were obtained using a specially designed bending machine under simple and oscillatory loading conditions, respectively. RESULTS: The TZP components submitted to cyclic loading in water exhibited subcritical crack propagation at stress levels significantly ( approximately 50%) lower than the critical stress intensity factor (K(IC)=5.6 MPam(1/2)). In spite of this susceptibility to subcritical crack growth, calculations based on the fatigue parameters and on the stress applied on the prosthesis indicate that posterior bridges with zirconia frameworks can exhibit lifetimes longer than 20 years if the diameter of the bridge connector is properly designed. SIGNIFICANCE: This in vitro study indicates that partially stabilized zirconia (3Y-TZP) can withstand the severe cyclic loading and wet conditions typically applied in the molar region of the mouth and is therefore an appropriate material for the fabrication of all-ceramic multi-unit posterior bridges. PMID- 16473403 TI - Mechanical and fracture behavior of veneer-framework composites for all-ceramic dental bridges. AB - OBJECTIVES: High-strength ceramics are required in dental posterior restorations in order to withstand the excessive tensile stresses that occur during mastication. The aim of this study was to investigate the fracture behavior and the fast-fracture mechanical strength of three veneer-framework composites (Empress 2/IPS Eris, TZP/Cercon S and Inceram-Zirconia/Vita VM7) for all-ceramic dental bridges. METHODS: The load bearing capacity of the veneer-framework composites were evaluated using a bending mechanical apparatus. The stress distribution through the rectangular-shaped layered samples was assessed using simple beam calculations and used to estimate the fracture strength of the veneer layer. Optical microscopy of fractured specimens was employed to determine the origin of cracks and the fracture mode. RESULTS: Under fast fracture conditions, cracks were observed to initiate on, or close to, the veneer outer surface and propagate towards the inner framework material. Crack deflection occurred at the veneer-framework interface of composites containing a tough framework material (TZP/Cercon S and Inceram-Zirconia/Vita VM7), as opposed to the straight propagation observed in the case of weaker frameworks (Empress 2/IPS Eris). SIGNIFICANCE: The mechanical strength of dental composites containing a weak framework (K(IC)<3 MPam(1/2)) is ultimately determined by the low fracture strength of the veneer layer, since no crack arresting occurs at the veneer framework interface. Therefore, high-toughness ceramics (K(IC)>5 MPam(1/2)) should be used as framework materials of posterior all-ceramic bridges, so that cracks propagating from the veneer layer do not lead to a premature failure of the prosthesis. PMID- 16473404 TI - Characterisation of electrospun polystyrene scaffolds for three-dimensional in vitro biological studies. AB - The purpose of this study was to produce a well-characterised electrospun polystyrene scaffold which could be used routinely for three-dimensional (3D) cell culture experimentation. A linear relationship (p<0.01) between three principal process variables (applied voltage, working distance and polymer concentration) and fibre diameter was reliably established enabling a mathematical model to be developed to standardise the electrospinning process. Surface chemistry and bulk architecture were manipulated to increase wetting and handling characteristics, respectively. X-ray photoelectron spectroscopy (XPS) confirmed the presence of oxygen-containing groups after argon plasma treatment, resulting in a similar surface chemistry to treated tissue culture plastic. The bulk architecture of the scaffolds was characterised by scanning electron microscopy (SEM) to assess the alignment of both random and aligned electrospun fibres, which were calculated to be 0.15 and 0.66, respectively. This compared to 0.51 for collagen fibres associated with native tissue. Tensile strength and strain of approximately of 0.15 MPa and 2.5%, respectively, allowed the scaffolds to be routinely handled for tissue culture purposes. The efficiency of attachment of smooth muscle cells to electrospun scaffolds was assessed using a modified 3 [4,5-dimethyl(thiazol-2yl)-3,5-diphery] tetrazolium bromide assay and cell morphology was assessed by phalloidin-FITC staining of F-actin. Argon plasma treatment of electrospun polystyrene scaffold resulted in significantly increased cell attachment (p<0.05). The alignment factors of the actin filaments were 0.19 and 0.74 for the random and aligned scaffold respectively, compared to 0.51 for the native tissue. The data suggests that electrospinning of polystyrene generates 3D scaffolds which complement polystyrene used in 2D cell culture systems. PMID- 16473405 TI - Autonomous regulation of free Ca2+ concentrations in isolated plant cell nuclei: a mathematical analysis. AB - Experiments performed on nuclei isolated from animal or plant cells have provided evidence that the nucleus generates directly specific nucleoplasmic calcium transients in response to external stimuli. Recent data suggest that isolated plant nuclei might be considered as a closed system where the nuclear concentration of free calcium would be regulated by reversible movements between the nucleoplasm and nuclear stores. We have addressed the relevance of this hypothesis by developing a mathematical approach to simulate nucleoplasmic calcium dynamics generated under various pH and temperature conditions. Here, we show that the experimental results could be explained provided that calcium channels as well as systems transporting calcium are present on the inner nuclear membrane. The putative channels would allow the entry of calcium into the nucleoplasm whereas the elusive transporting system(s) would contribute to replenish the nuclear stores. The simple proposed model is versatile enough to explain and predict autonomous changes in free calcium in the nucleoplasm of isolated plant nuclei. PMID- 16473407 TI - Construction of a conventional non-radioisotope method to quantify HM1.24 antigens: correlation of HM1.24 levels and ADCC activity of the humanized antibody against HM1.24. AB - A humanized monoclonal antibody (mAb) against HM1.24 (AHM) caused antibody dependent cellular cytotoxicity (ADCC) against multiple myeloma (MM) cells. Here, we constructed a conventional non-radioisotope method that quantifies the amount of HM1.24 using fluorescein-labeled AHM. More than 10(4) molecules/cell of HM1.24 were detected in 12 out of 14 patients' MM cells, and a linear correlation was found between ADCC by AHM and the amounts of HM1.24. Thus, AHM is likely to be more efficacious against MM cells with high levels of HM1.24. This conventional non-RI method to quantify HM1.24 will be useful to select patients most likely to respond to AHM. PMID- 16473406 TI - ATRA can enhance apoptosis that is induced by Flt3 tyrosine kinase inhibition in Flt3-ITD positive cells. AB - Among activating Flt3 mutations that have been shown in 25-30% of acute myeloid leukaemia (AML) Flt3-internal tandem duplication (ITD) mutations are predominant. We investigated the influence of all-trans-retinoic acid (ATRA) and granulocyte colony stimulating factor (G-CSF) for their effects on differentiation and apoptosis in human cell lines with different Flt3 variants (THP-1 versus MV4-11 and MOLM13) dependent on the inhibition of Flt3 tyrosine kinase by the bis(lH-2 indolyl)methanone D-65476. While myeloid differentiation was not observed in both Flt3-ITD cell lines (MV4-11 and MOLM13), we demonstrate an enhanced proapoptotic effect of D-65476 in the presence of ATRA that was restricted to the Flt3-ITD expressing cells. The combined treatment with ATRA and D-65476 also led to a pronounced down-regulation of surviv in on mRNA and protein level in Flt3-ITD but not in Flt3 wildtype expressing cells (THP-1). Surprisingly, there was no differential expression of important proteins like Bcl-X(L), Bcl-2 or Bax that might explain enhanced apoptosis. Furthermore, Akt phosphorylation after stimulation with Flt3 ligand dependent on D-65476 was not affected by pretreatment with ATRA. We suggest that regulation of inhibitors of apoptosis might play a crucial role how ATRA can increase the proapoptotic effect of Flt3 inhibitors in myeloid leukemia cells expressing Flt3-ITD. This effect can potentially be exploited for the treatment of Flt3-ITD positive acute myeloid leukemia. PMID- 16473409 TI - Visceral leishmaniasis complicated by fungal pulmonary valve endocarditis. AB - We present a rare neglected case of fungal pulmonary valve endocarditis which presented with typical extra cardiac manifestations after repeated injections for treatment of visceral leishmaniasis. Surgical intervention to replace the pulmonary valve was the only option to manage the patient in spite of extensive medical treatment. PMID- 16473408 TI - Prevalence of hepatitis C virus infection in children admitted to an urban hospital. AB - BACKGROUND AND AIM: Hepatitis C infection is a global health problem affecting about 3% of the world's population. However, very little data exists on the prevalence of hepatitis C virus infection in childhood. METHODS: We performed a cross-sectional study in 2000 children and adolescents who were treated as in- or out patients in our hospital. Blood samples were collected between February 2002 and June 2004 and were tested for HCV antibodies (anti-HCV). Positive samples were further investigated by HCV specific PCR and Western blot assay. RESULTS: Mean age of children was 8.1 years. 908 (45%) were female and 1092 (55%) male. One thousand eight hundred and ninety-eight were Caucasian, 37 African, and 65 Asian. 16 (0.8%) tested positive for anti-HCV. HCV-RNA was detectable in one child (0.05%), and three were positive in the Western blot assay (0.15%). The HCV viremic child had received multiple blood transfusions after cardiac surgery. She was asymptomatic with normal transaminases. Seroprevalence of HCV antibodies were equally distributed among boys and girls. CONCLUSIONS: The prevalence of persistent hepatitis C in children from an urban hospital in Germany is low. Most patients with HCV antibodies are not infected. Therefore, although universal screening is not warranted, it should always be performed in risk groups such as transfused children because HCV infection in childhood is usually asymptomatic. PMID- 16473410 TI - Characterisation of rickettsial diseases in a hospital-based population in Malta. AB - OBJECTIVE: The aim of the study was to characterise the causative agents of rickettsial disease in Malta. A secondary objective was to study the epidemiology of cases of rickettsial disease. METHODS: Cases admitted to St Luke's Hospital between June 2002 and May 2003 presenting with complaints of fever, headache, rash and/or an eschar were considered possible cases of rickettsial disease. A patient interview was conducted within 24h of admission. Paired sera were taken for serology and blood samples sent for rickettsial PCR and culture. Whenever an eschar was present, biopsies were taken for culture and immunohistochemical analysis. RESULTS: Thirty-three cases of possible rickettsial disease were identified. Although serological tests showed cross reactivities between different species of rickettsiae, one was diagnostic for Rickettsia conorii. None of the sera showed any cross-reactivity with Rickettsia typhi. There was one positive biopsy for R. conorii when tested by PCR and another was positive for spotted fever group Rickettsia by immunohistochemistry. CONCLUSION: Spotted fever rickettsiosis is endemic in Malta. Contrary to previous belief, none of the cases were due to murine typhus. The predominant causative agent of rickettsial disease in Malta is likely to be R. conorii, although the animal reservoir has still not been definitely identified. PMID- 16473411 TI - Estrogen therapy and thrombotic risk. AB - Post-menopausal hormone therapy increases the risk for venous thrombosis, and possibly myocardial infarction (MI) and ischemic stroke. However, most women using hormone therapy do not suffer thrombosis, and to date our ability to identify women at risk is limited. Thrombosis, arterial or venous, has 2 requisites: a vascular anomaly and a response of the hemostasis system to the anomaly. Consequently, experimental approaches to understand the pathophysiology of thrombosis require definition of vascular anatomy and function as well as characteristics of the blood within the context of genetic background, lifestyle choices and environmental exposures, which influence gene expression. Defining interactions among factors that affect individual propensity to thrombosis will allow physicians to better identify at-risk individuals, for example a woman contemplating estrogen therapy for symptoms of menopause, and prevent adverse thrombotic events. PMID- 16473412 TI - Effects of Salmonella enterica serovars Typhimurium (ST) and Choleraesuis (SC) on chemokine and cytokine expression in swine ileum and jejunal epithelial cells. AB - The gastrointestinal epithelium represents a barrier to potentially invasive enteric pathogens, maintains a role in innate immune surveillance, and is a source of both chemokine and cytokine chemotactic mediators in response to bacterial invasion. In the current study, we evaluated cytokine and chemokine mediators known to regulate movement of macrophages (macrophage migration inhibitory factor; MIF), neutrophils (IL8), dendritic cells (CCL20), and epithelial remodeling (osteopontin; OPN) in response to invasive swine enteropathogens Salmonella enterica serovar Typhimurium (ST) or Choleraesuis (SC). For the in vivo experiment, weaned pigs served as uninfected controls (0 h) or were given 3 x 10(9) CFU ST orally. Pigs were sacrificed at 8, 24, 48, and 144 h after inoculation and total RNA was extracted from defined segments of proximal (PI) and distal (DI) ileum. Relative expression of MIF and OPN were not affected by ST. IL8 expression was increased numerically (P = 0.17 for the interaction term) at 24 and 144 h in the PI and these increases accounted for greater expression in the PI relative to the DI (P < 0.05). Relative expression of CCL20 was increased at 24 h after ST (P < 0.05). Next, we evaluated the time course of MIF, IL8, CCL20, and OPN mRNA expression induced by application of lipopolysaccharide (LPS), ST or SC in vitro using pig jejunal epithelial cells (IPEC-J2). Cells were grown to confluency on permeable membranes, and treated apically with LPS (10 ng/mL), ST or SC (10(8)/well). After 1 h, cells were washed to remove LPS or extracellular bacteria, and media containing gentamicin was added to kill remaining extracellular bacteria. Media and RNA were collected at 1.5, 3, and 6 h after treatment. MIF mRNA was not affected by LPS or bacterial treatment. Similarly, IL8 expression was not affected by LPS, but was increased by ST and SC relative to controls at 1.5 and 3 h post exposure (P < 0.05 for all comparisons). Treatment with SC increased CCL20 mRNA relative to controls at 3 h (P < 0.05), while ST increased CCL20 at 1.5, 3, and 6h with maximal expression at 6 h (P < 0.05 for all comparisons). ST and SC increased polarized IL8 secretion. Our data demonstrate that invasive bacterial pathogens in the pig gastrointestinal tract trigger upregulation of selected cytokine and chemokine mediators, but serovars of Salmonella elicited differing patterns of activation in vitro. PMID- 16473413 TI - Cytokine responses to EHV-1 infection in immune and non-immune ponies. AB - Protecting equids against equine herpesvirus-1 (EHV-1) infection remains an elusive goal. Repeated infection with EHV-1 leads to protective immunity against clinical respiratory disease, and a study was conducted to measure the regulatory cytokine response (IFN-gamma and IL-4) in repeatedly infected immune ponies compared to non-immune ponies. Two groups of four ponies were established. Group 1 ponies had previously been infected on two occasions, and most recently 7 months before this study. Group 2 ponies had no history no vaccination or challenge infection prior to this study. Both groups were subjected to an intranasal challenge infection with EHV-1, and blood samples were collected pre infection, and at 7 and 21 days post-infection for preparation of PBMCs. At each time point, the in vitro responses of PBMCs to stimulation with EHV-1 were measured, including IFN-gamma and IL-4 mRNA production, and lymphoproliferation. Group 1 ponies showed no signs of clinical disease or viral shedding after challenge infection. Group 2 ponies experienced a biphasic pyrexia, mucopurulent nasal discharge, and nasal shedding of virus after infection. Group 1 ponies had an immune response characterized both before and subsequent to challenge infection by an IFN-gamma response to EHV-1 in the absence of an IL-4 response, and demonstrated increased EHV-1-specific lymphoproliferation post-infection. Group 2 ponies had limited cytokine or lymphoproliferative responses to EHV-1 pre challenge, and demonstrated increases in both IFN-gamma and IL-4 responses post challenge, but without any lymphoproliferative response. Protective immunity to EHV-1 infection was therefore characterized by a polarized IFN-gamma dependent immunoregulatory cytokine response. PMID- 16473415 TI - Smooth muscle research: from Edith Bulbring onwards. AB - The properties of smooth muscle are currently being studied extensively. Indeed, the small size of myocytes and the huge range of behaviours they exhibit make them an attractive focus for current research. However, this was not always the case. These properties initially made smooth muscles more difficult to study than the larger specialized striated muscles that were the focus of attention of leading researchers. In the UK, research into the physiology of smooth muscles began in the Pharmacology Department at Oxford, led by Edith Bulbring; her early results attracted much attention and resulted in the formation of an active international smooth muscle research group. Although several areas of current interest in the field of smooth muscle were not tackled by the Oxford group, progression of much of the field has clear links to Bulbring and her group. PMID- 16473414 TI - Equine herpesvirus 1 glycoprotein D expressed in E. coli provides partial protection against equine herpesvirus infection in mice and elicits virus neutralizing antibodies in the horse. AB - The envelope glycoprotein D of EHV-1 (EHV-1 gD) is essential for virus infectivity and entry of virus into cells and is a potent inducer of virus neutralizing antibody. In this study, truncated EHV-1 gD (gDt) was expressed with a C-terminal hexahistidine tag in E. coli using a pET vector. Western blot analysis using an anti-gD monoclonal antibody demonstrated the presence of gDt bands at 37.5, 36, 29.5 and 28 kDa. The immunogenicity and protective efficacy of partially purified gDt was compared with gD expressed in insect cells by a recombinant baculovirus (Bac gD) using a BALB/c mouse model of EHV-1 respiratory infection. The proteins were also compared in a prime-boost protocol following an initial inoculation with gD DNA. gDt elicited similar levels of gD-specific antibody and neutralizing antibody compared with Bac gD and also provided a similar level of protection against EHV-1 challenge in mice. Inoculation of horses with gDt elicited EHV-1 gD-specific antibodies including virus neutralizing antibody, suggesting that despite the lack of glycosylation, E. coli may be a useful vehicle for large scale production of EHV-1 gD for vaccine studies. PMID- 16473416 TI - Inhibition of human immunodeficiency virus type 1 infection in macrophages by an alpha-v integrin blocking antibody. AB - Macrophages are key cells for HIV infection and HIV spreading inside the organism. Macrophages cultured in vitro can be successfully infected after differentiation with cytokines such as macrophage colony stimulating factor (M CSF). In the monocyte to macrophage differentiation process with M-CSF, alphav integrins are upregulated concomitantly with the capacity of HIV to generate a productive virus infection. In the present study we show that an anti-alphav antibody, 17E6, inhibited HIV-1 infection of primary macrophages. The effect of 17E6 on HIV-1 BaL replication in acutely infected macrophages was dose-dependent, with a 50% effective concentration (EC50) of 17+/-2 microg/ml in the absence of cytotoxicity. Similarly, a monoclonal antibody targeting the alphavbeta6 integrin (14D9.F8) also inhibited HIV-1 BaL infection in this cell type. 17E6 reduced the detection of HIV-1 BaL proviral DNA in acutely infected macrophages, but was completely ineffective against HIV-1 BaL production in chronically infected macrophages, suggesting that 17E6 inhibited HIV infection at an early stage of the virus cycle. Finally, a small molecular weight antagonist of the alphavbeta6 integrin, EMD 409849, reduced HIV replication at subtoxic concentrations. Therefore, our results suggest that alphav-containing integrins could play a role in HIV replication in macrophages and suggest that small-molecular-weight compounds might interfere with HIV replication in macrophages through the interaction with alphav integrins. PMID- 16473417 TI - Antiviral activity and cross-resistance profile of P-1946, a novel human immunodeficiency virus type 1 protease inhibitor. AB - The HIV protease inhibitor P-1946 is a member of a novel family of l-Lysine derivatives. The compound is a specific HIV-1 protease inhibitor that has potent and selective in vitro antiviral activity (EC50 152 nM) against a range of isolates resistant to commercially available protease inhibitors. The presence of at least four primary and four secondary drug resistance mutations is required to achieve greater than four-fold resistance to P-1946. P-1946's favorable resistance profile makes it a good lead for the development of new agents active against existing PI-resistant virus in treatment-experienced patient. PMID- 16473418 TI - Environmental enrichment increases responding to contextual cues but decreases overall conditioned fear in the rat. AB - This study aimed at investigating the effects of environmental enrichment on various aspects of contextual processing in adult female rats. In experiment 1, simple conditioning was studied using either a training procedure allowing overshadowing of the contextual cues by signalling footshock with a discrete tone or a training procedure allowing a reduction of this overshadowing by explicitly unpairing the footshock and the tone. In experiment 2, contextual discrimination and contextual occasion-setting were assessed. Rats were daily exposed to two different contexts. In one context, a footshock was delivered 30s after the offset of a tone, whereas in the other context the same tone was presented alone. Experiment 3 examined familiarization to a new context. Experiment 1 showed that environmental enrichment reduced the overshadowing of contextual cues by the tone and also reduced freezing to the more predictive cue according to the training procedure used. Experiment 2 showed that environmental enrichment increased the ability of rats to discriminate two contexts. Experiment 3 showed that enriched rats familiarized faster to a new context than standard rats. Taken together, these results suggest that environmental enrichment in adult rats enhances learning about contextual cues and reduces overall fear associated with aversive events. PMID- 16473419 TI - Safrole oxide induced human umbilical vein vascular endothelial cell differentiation into neuron-like cells by depressing the reactive oxygen species level at the low concentration. AB - Previously, we found that 5-25 microg/ml safrole oxide could inhibit apoptosis and dramatically make a morphological change in human umbilical vein vascular endothelial cells (HUVECs). But the possible mechanism by which safrole oxide function is unknown. To answer this question, in this study, we first investigated the effects of it on the activity of nitric oxide synthetase (NOS), the expressions of Fas and integrin beta4, which play important roles in HUVEC growth and apoptosis, respectively. The results showed that, at the low concentration (10 microg/ml), safrole oxide had no effects on NOS activity and the expressions of Fas and integrin beta4. Then, we investigated whether HUVECs underwent differentiation. We examined the expressions of neuron-specific enolase (NSE) and neurofilament-L (NF-L). Furthermore, we analyzed the changes of intracellular reactive oxygen species (ROS). After 10 h of treatment with 10 microg/ml safrole oxide, some HUVECs became neuron-like cells in morphology, and intensively displayed positive NSE and NF-L. Simultaneously, ROS levels dramatically decreased during HUVECs differentiation towards neuron-like cells. At the low concentration, safrole oxide induced HUVECs differentiation into neuron-like cells. Furthermore, our data suggested that safrole oxide might perform this function by depressing intracellular ROS levels instead of by affecting cell growth or apoptosis signal pathways. PMID- 16473420 TI - A case-control study of factors associated with pelvic and tibial stress fractures in Thoroughbred racehorses in training in the UK. AB - Few epidemiological studies have investigated risk factors for musculoskeletal injury occurring in Thoroughbred racehorses during training, although it is the major cause of wastage in the racing industry. We recently conducted a large scale epidemiological study to estimate the incidence of fracture in racehorses in training in the UK and to identify associated risk factors. Thirteen racehorse trainers provided data on horses in their care, with daily recording of training information and provision of details on any fractures incurred. Data were collected for 2 years, including two consecutive flat racing seasons (1999 and 2000). This paper describes findings from a nested case-control study investigating factors associated with the occurrence of pelvic and tibial stress fractures in our study population. Cases were identified from the main study and defined as horses with a pelvic or tibial stress fracture, confirmed through routine diagnostic imaging. Randomly selected controls were matched on date of fracture in the case. Age and gender of the horse, its exercise history and training surfaces were examined as explanatory variables. Exercise was quantified as cumulative distances cantered and worked at high speed in 30- and 60-day periods prior to date of fracture in the case. Conditional logistic regression was used to construct multivariable models for the 30- and 60-day periods, respectively. We hypothesised that larger cumulative exercise distances would be associated with an increased risk of pelvic or tibial stress fracture and that different training surfaces would be associated with differences in fracture risk. In the 30-day period, when adjusting for trainer, the risk of pelvic or tibial stress fracture increased with increasing distance cantered, reaching a peak at around 50 km, after which the risk reduced. This trend was not obvious in the 60-day period, with no significant association between exercise distances and risk of stress fracture. Predominant use of one particular sand-based all-weather surface was related to an increased risk of pelvic or tibial stress injury, although this finding should be interpreted with caution. Surface maintenance and construction may play a role; variables that were not considered in the current analyses, which were based on a relatively small number of cases. Trainer was associated with differences in stress fracture risk after adjusting for exercise distances and surface but age and gender were not. PMID- 16473421 TI - Diversity, distribution and divergence of lin genes in hexachlorocyclohexane degrading sphingomonads. AB - Two forms of hexachlorocyclohexane (HCH), gamma-HCH (lindane) and technical HCH (incorporating alpha-, beta-, gamma- and delta- isomers), have been used against agricultural pests and in health programs since the 1940s. Although all the isomers are present in the milieu, delta- and beta-HCH isomers are the most problematic and present a serious environmental problem. Bacteria that degrade HCH isomers have been isolated from HCH contaminated soils from different geographical locations around the world (from the family Sphingomonadaceae). Interestingly, all these bacteria contain nearly identical lin genes (responsible for HCH degradation), which are diverging to perform several catabolic functions. The organization and diversity of lin genes have been studied among several sphingomonads, and they have been found to be associated with plasmids and IS6100, both of which appear to have a significant role in their horizontal transfer. The knowledge of the molecular genetics, diversity and distribution of lin genes, and the potential of sphingomonads to degrade HCH isomers, can now be used for developing bioremediation techniques for the decontamination of HCH contaminated sites. PMID- 16473422 TI - Mineralocorticoid and glucocorticoid receptor expressions in astrocytes and microglia in the gerbil hippocampal CA1 region after ischemic insult. AB - In the present study, we observed expression and changes of mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) in the gerbil hippocampal CA1 region, but not in the CA2/3 region, after 5 min of transient forebrain ischemia. In blood, corticosterone levels were increased biphasically at 30 min and 12 h after ischemia/reperfusion, and thereafter its levels were decreased. In the sham operated group, MR and GR immunoreactivities were weakly detected in the CA1 region. By 3 days after ischemia, MR and GR were not significantly altered in the CA1 region: at 12 h after ischemia, GR was expressed in a few neurons in the CA1 region, whereas MR was not expressed in any neurons after ischemic insult. From 4 days after ischemia, MR and GR immunoreactivities were detected in astrocytes and microglia in the CA1 region, and at 7 days after ischemia, MR and GR immunoreactivities peaked in the hippocampal CA1 region. At this time, 55% of astrocytes and 30% of microglia showed MR immunoreactivity, and 20% of astrocytes and 40% of microglia showed GR immunoreactivity. Western blot analyses showed that the pattern of changes in MR and GR protein levels was similar to the immunohistochemical changes observed after transient forebrain ischemia. From 4 days after ischemia, MR and GR protein levels were increased time-dependently after ischemia. In conclusion, enhanced MR and GR expressions in astrocytes and microglia were detected in the hippocampal CA1 region 4-7 days after ischemia/reperfusion. At this time, GR immunoreactivity was abundant in microglia, whereas MR immunoreactivity was prominent in astrocytes. The specific distribution of corticosteroid receptors in the astrocytes and microglia may be associated with the differences of MR and GR functions against ischemic damage. PMID- 16473423 TI - Visual cortical contribution to open-loop and feed-back control of convergence eye movements in the cat. AB - Open-loop and closed loop controls in convergence eye movement have been reported by human psycho-physical studies. To investigate the visual cortical involvement in open-loop convergence eye movement, we trained a cat to elicit anticipatory convergence (convergence eye movement before the onset of target movement) by using an approaching visual target with a preceding alarm signal. After 1-2 weeks of training, anticipatory convergence was observed in more than half of the trials in seven cats. The frequency of occurrence of anticipatory convergence was significantly decreased after electrocoagulation in the convergence-related region of the lateral suprasylvian (LS) area, an extrastriate visual cortex of the cat. On the other hand, the localized injection of Muscimol, a GABA-A agonist, reduced visually evoked convergence, but caused no significant effects to anticipatory convergence. These differential results suggest that the LS plays a role in both open-loop and feed-back control of convergence eye movement, and a GABA-A sensitive subregion is involved in the feed-back control of convergence eye movement. PMID- 16473424 TI - Zaltoprofen, a non-steroidal anti-inflammatory drug, inhibits bradykinin-induced pain responses without blocking bradykinin receptors. AB - Zaltoprofen, a preferential COX-2 inhibitor, exhibited a potent inhibitory action on the nociceptive responses induced by a retrograde infusion of bradykinin into the right common carotid artery in rats. However, other COX-2 preferential inhibitors such as meloxicam and etodolac did not exhibit any apparent action, and also, preferential COX-1 inhibitors mofezolac and indomethacin, COX-1 and COX 2 inhibitor loxoprofen sodium showed a weak effect. These non-steroidal anti inflammatory drugs (NSAIDs) except for zaltoprofen, strongly inhibited an acetic acid-induced writhing response related to PGs based on COX-1, at lower doses. Zaltoprofen had a moderate inhibitory effect compared with those of the above mentioned NSAIDs. These results suggest that the inhibitory effect of zaltoprofen on bradykinin-induced nociceptive responses is not explainable by the inhibition of cyclooxygenase (COX). So, we examined the inhibitory effect of zaltoprofen on bradykinin-induced nociceptive responses by performing several in vitro experiments. Zaltoprofen did not bind to B(1) and B(2) receptors in a radio ligand binding assay. In the cultured dorsal root ganglion cells of mature mice, zaltoprofen completely inhibited the bradykinin-induced increase of [Ca(2+)](i), which was inhibited by B(2) antagonist D-Arg-[Hyp(3), Thi(5,8), D-Phe(7)] bradykinin, but not by B(1) antagonist. In addition, the inhibition of zaltoprofen on the increase of [Ca(2+)](i) was observed even under extracellular Ca(2+)-free conditions. The above results suggest that zaltoprofen produces an analgesic action on bradykinin-induced nociceptive responses by blocking the B(2) receptor-mediated pathway in the primary sensory neurons. Taken together, these results suggest that zaltoprofen may serve as a potent and superior analgesic for the treatment of pain. PMID- 16473425 TI - Changes in protein synthesis during thermal adaptation of Propionibacterium freudenreichii subsp. shermanii. AB - Dairy propionibacteria are present in Graviera Kritis, a traditional Gruyere-type cheese made without added propionic starter. Ten isolated strains were identified by a combination of SDS-PAGE, species-specific PCR and according to their ability to ferment lactose. They were all found to belong to the Propionibacterium freudenreichii subsp. shermanii species. Because of the stressing Gruyere technology, which includes cooking at 52 to 53 degrees C their thermotolerance was investigated at 55 degrees C. Thermotolerant and thermosensitive strains were clearly discriminated. Interestingly, the reference strain CIP 103027 belongs to the sensitive subset. One sensitive strain, ACA-DC 1305 and one tolerant, ACA-DC 1451, were selected for further study and compared to CIP 103027. For the sensitive strains ACA-DC 1305 and CIP 103027, heat pre-treatment at 42 degrees C conferred thermoprotection of cells at the lethal temperature of 55 degrees C, while there was less effect on the tolerant ACA-DC 1451. No cross-protection of salt-adapted cells against heat stress was observed for none of the strains. Differential proteomic analysis revealed distinct but overlapping cell responses to heat stress between sensitive and tolerant strains. Thermal adaptation upregulated typical HSPs involved in protein repair or turnover in the sensitive one. In the tolerant one, a distinct subset of proteins was overexpressed, whatever the temperature used, in addition to HSPs. This included enzymes involved in propionic fermentation, amino acid metabolism, oxidative stress remediation and nucleotide phosphorylation. These results bring new insights into thermoprotection in propionibacteria and the occurrence of divergent phenotypes within a same subspecies. PMID- 16473426 TI - Persistence of caliciviruses on environmental surfaces and their transfer to food. AB - The noroviruses (NoV) are a common cause of human gastroenteritis whose transmission by foodborne routes is well documented. Fecally contaminated surfaces are likely to contribute to this foodborne transmission and to the propagation of viral disease outbreaks. The purpose of this study was to (i) investigate the stability of NoV on various food preparation surfaces; and (ii) evaluate the degree of virus transfer from these surfaces to a model-ready-to-eat (RTE) food. For the virus persistence experiments, stainless steel, formica and ceramic coupons were artificially contaminated with Norwalk virus (NV), the prototype genogroup I NoV; NV RNA; or feline calicivirus (FCV) F9 (a NoV surrogate), stored at ambient temperature for up to 7 d, and periodically assayed for detection. In the transfer experiments, stainless steel coupons were inoculated with NV or FCV F9 and allowed to dry for 10, 30 and 60 min, after which lettuce leaves were exposed to the surface of the coupons at various contact pressures (10, 100, and 1000 g/9 cm2). Virus recovery was evaluated by RT PCR (for NV and NV RNA) or by plaque assay (for FCV F9) using Crandell Reese Feline Kidney (CRFK) cells. NV and FCV were detected on all three surfaces for up to 7 d post-inoculation; for FCV, there was an approximate 6 to 7-log10 drop in virus titer over the 7 d evaluation period. By contrast, when stainless steel was inoculated with purified NV RNA, RT-PCR detection was not possible beyond 24 h. Transfer of both NV and FCV from stainless steel surfaces to lettuce occurred with relative ease. This study confirms lengthy NoV persistence on common food preparation surfaces and their ease of transfer, confirming a potential role for environmental contamination in the propagation of viral gastroenteritis. PMID- 16473427 TI - Effects of mechanical stress on Anammox granules in a sequencing batch reactor (SBR). AB - The effect of shear stress on Anammox process was studied in a sequencing batch reactor (SBR). The reactor was operated during 218 days under different stirring speeds (60-250 rpm) in order to expose the system to different shear conditions and to study the stability of the Anammox granules referred to their biological activity and size. The nitrogen loading rate (NLR) fed to the SBR was kept around 0.3g N(L day)(-1). The nitrite (limiting substrate) removal percentage was 98% during most of the operational period. The specific Anammox activity of the biomass was practically constant and around 0.4 g N(g VSSday)(-1) and the average feret diameter of the formed granules was 0.64 mm. Obtained results indicated that stirring speeds up to 180 rpm have no negative effect on the performance of the Anammox process, whereas Anammox activity decreased to 40% when a rotating speed of 250 rpm was tested and the average diameter decreased in 45%, the concentration of solids in the effluent increased to 0.2g TSSL(-1) and nitrite was accumulated in the reactor up to 60 mg NL(-1). PMID- 16473428 TI - Infectious hypodermal and hematopoietic necrosis virus (IHHNV)-related sequences in the genome of the black tiger prawn Penaeus monodon from Africa and Australia. AB - We found an infectious hypodermal and hematopoietic necrosis virus (IHHNV) related sequence within the shrimp genome in populations of Penaeus monodon from Africa and Australia. IHHNV is a single-stranded DNA virus that has caused severe mortality and stunted growth in penaeid shrimp. Recently, IHHNV-related sequences were found in samples of P. monodon from Madagascar and Tanzania. These sequences vary considerably (14 and 8%, respectively) from that of IHHNV found in association with viral epidemics. Laboratory bioassays were carried out with P. monodon and Litopenaeus vannamei to determine if either of these IHHNV-related sequences is infectious. We used juvenile and adult P. monodon containing the virus-related sequences from four geographic regions to generate inocula and tissues for feeding. Specific pathogen free P. monodon and L. vannamei were used as indicator shrimp. During the 2-4 week bioassays, none of the indicator shrimp showed signs of infection or disease. Results of both PCR assays and histological examination of the indicator shrimp were negative for IHHNV infection, indicating that the Africa type IHHNV-related sequences are not infectious. With the shrimp containing the Madagascar type IHHNV-related sequence (designated as type A), we performed genome walking at the 3' end of the virus-related sequence and found that this virus-related sequence is part of the P. monodon genome. A fragment of 1.9 kb flanking sequence was cloned and sequenced. Sequence analysis showed that this flanking sequence contains shrimp microsatellite DNA. Also, its translated amino acid sequence was highly similar to a retrotransposon. This result provides molecular evidence that the type A IHHNV-related sequence is shrimp DNA. This sequence was found in the P. monodon collected from Africa and Australia. PMID- 16473429 TI - A highly attenuated rabies virus HEP-Flury strain reverts to virulent by single amino acid substitution to arginine at position 333 in glycoprotein. AB - An amino acid at position 333 in the glycoprotein of several fixed rabies virus strains is responsible for the pathogenicity in adult mice. Substitution of arginine at this position largely reduces the viral pathogenicity in adult mice. Attenuation by this single amino acid substitution has been established by using escape mutants selected by monoclonal antibodies and point-mutated virus generated by reverse-genetics. A highly attenuated HEP-Flury strain, which was selected by serial passages in cell cultures, has glutamine at this position. In this study, a point-mutated rHEP333R virus, having arginine at position 333, was generated and examined for the responsibility of this substitution in rabies pathogenicity. The rHEP333R acquired an ability to spread and propagate in mouse brain but the parental rHEP did not. The pathogenicity of rHEP333R to adult mice by intracerebral inoculation largely increased. We confirmed that an arginine at position 333 contributed to reversion of the pathogenicity in a highly attenuated HEP-Flury strain. PMID- 16473430 TI - Development of an oral sustained release drug delivery system utilizing pH dependent swelling of carboxyvinyl polymer. AB - A new oral sustained-release (SR) drug delivery system utilizing pH-dependent swelling of carboxyvinyl polymer (CP) has been proposed. This swelling polymer incorporation layer system, referred to as SPILA system, consists of core granules coated with a mixture of CP, water insoluble polymer, and water-soluble polymer (WP). Release profiles of metoprolol tartrate (ME) from SPILA system were pH dependent: drug release was slower in the medium of pH 1.2 than in the medium of pH 6.8 due to a coating layer with pH-dependent swelling polymer, CP. Lower C(max), longer T(max), longer MRT and higher AUC values were obtained following administration of SR granules based on SPILA system to beagle dogs compared with pH-independent SR granules having a coating layer without CP, while both SR granules provided similar in vitro release profiles. Moreover, using morphine hydrochloride (MO), in vitro and in vivo performances of the SPILA system were investigated. pH-dependency on the release profiles of MO from SPILA system was more evident when the amount of CP incorporated in SPILA system was increased. AUC and MRT values following administration of SR granules of MO with a coating layer containing 8% of CP to beagle dogs were 191 ng h/mL and 10.6 h respectively, while those following SR granules of MO with a coating layer containing 1% of CP to beagle dogs were 86.4 ng h/mL and 5.86 h, respectively. An important role of CP in the release-regulating layer of SPILA system for keeping the higher and more extended plasma levels of morphine free base was confirmed from the in vivo performance of SPILA system. PMID- 16473431 TI - Occult HBV infection and blood transfusion. PMID- 16473433 TI - Complete weaning off immunosuppression in HCV liver transplant recipients is feasible and favourably impacts on the progression of disease recurrence. AB - BACKGROUND/AIMS: HCV-related disease recurrence progresses rapidly after liver transplantation. We hypothesised that withdrawal of immunosuppression might favourably impact on disease progression. METHODS: Weaning off immunosuppression was attempted in 34 HCV-RNA positive patients (mean age 62+/-6.4 years) transplanted 63.5+/-20.1 months earlier, under cyclosporine A monotherapy. Patients were followed for 3 years including yearly protocol liver biopsies. Primary endpoints were feasibility of weaning off immunosuppression and its impact on disease progression. Secondary endpoint was to identify predictors of an immunosuppression-free state and fibrosis progression. RESULTS: Complete and permanent immunosuppression withdrawal was achieved in 8 patients (23.4%), whereas 14 (41.2%) developed rejection within eight months despite an initial response and 12 (35.2%) rejected during tapering. After a mean follow-up 45.5+/ 5.8 months weaned patients showed stabilisation/improvement of histological fibrosis (P<0.01), lower necro-inflammation (P<0.02) and improved liver function (P<0.05) compared to weaning-intolerants. Multiple logistic regression identified low blood cyclosporine A trough levels during the first post-transplant week (P=0.004) and initial steroid-free immunosuppression (P<0.008) as independent predictors of sustained weaning. Achievement of immunosoppression freedom (P=0.02) and baseline staging score (P<0.0001) were independently associated with stabilisation/improvement of histological fibrosis. CONCLUSIONS: Reconstitution of immune-competence in the host improves the natural history of HCV recurrence in the graft. PMID- 16473434 TI - New localism in the English National Health Service: what is it for? AB - This paper discusses the decentralisation in the English National Health Service (and the new organisational form of Foundation Trusts, in particular) in the context of the trend for decentralising public services generally since the New Labour government started its second term of office in 2001. It considers the likelihood of decentralisation achieving the aims policy makers have set: these are better services and an increase in local people's participation in the planning and running of services (active citizenship). Looking at the evidence to date, both about decentralisation of health services and about Foundation Trusts, the achievement of these goals seems uncertain. PMID- 16473435 TI - The diffusion of medical technology, local conditions, and technology re invention: a comparative case study on coronary stenting. AB - Innovation of medical technology is a major driving force behind the increase in medical expenditures in developed countries. Previous studies identified that the diffusion of medical technology varied across countries according to the characteristics of regulatory policy and payment systems. Based on Roger's diffusion of innovation theory, this study purported to see how local practice norms, the evolving nature of diffusing technology, and local clinical needs in addition to differences in politico-economic systems would affect the process of innovation diffusion. Taking a case of coronary stenting, an innovative therapeutic technology in early 1990s, we provided a case study of hospital-based data between two teaching high-tech hospitals in Japan and the US for discussion. Stenting began to be widely used in both countries when complementary new technology modified its clinical efficacy, but the diffusion process still differed between the two hospitals due to (1) distinctive payment systems for hospitals and physicians, (2) practice norms in favor of percutaneous intervention rather than bypass surgery that was shaped by payment incentives and cultural attitudes, and (3) local patient's clinical characteristics that the technology had to be tailored for. The case study described the diffusion of stent technology as a dynamic process between patients, physicians, hospitals, health care systems, and technology under global and local conditions. PMID- 16473436 TI - Do policy changes in the pharmaceutical reimbursement schedule affect drug expenditures? Interrupted time series analysis of cost, volume and cost per volume trends in Sweden 1986-2002. AB - The last decades increasing pharmaceutical expenditures in Sweden and other western countries have created a need for reforms to reduce the trend. The aim was to analyse if reforms concerning the pharmaceutical reimbursement scheme in Sweden during the years 1986-2002 were associated with changes in cost, volume and cost per volume of pharmaceuticals. Effects of changes in the reimbursement schedule during the study period were evaluated for all registered pharmaceuticals in Sweden and for five indicator drug groups. Five policy changes during the study period were assessed. Three concerned increased patient co payment (January 1, 1991; January 1, 1995 and June 1, 1999), one the introduction of reference based pricing and increased co-payment (January 1, 1993) and one a new structure of the reimbursement schedule (January 1, 1997). The National Corporation of Swedish Pharmacies provided pharmaceutical delivery data for all Swedish pharmacies. Possible breaks in the trend associated with the investigated reforms were analysed with linear segmented regression analysis. This showed that increased co-payments were not associated with changed level or slope of cost and volume. The new reimbursement schedule was associated with a decreased level of cost and volume, both for all drugs combined and for several of the indicator drug groups. It was also associated with an increased slope for both volume and cost in some indicator drug groups and for all drugs. Introduction of reference based pricing was associated with a reduced slope of cost/defined daily doses (DDD) in all of the indicator drug groups and for all drugs. The analysis showed that major changes in the reimbursement system such as the introduction of a new reimbursement schedule and reference based pricing were associated with reductions in cost and volume for the new reimbursement schedule and cost per volume for reference based pricing. PMID- 16473437 TI - Interprovincial migration of physicians in Canada: where are they moving and why? AB - This paper analyzes the provincial characteristics contributing to physician's decision in his/her choice of province of residence. It (1) attempts to determine if, and the extent to which, expected income in a province plays a role in physician's decision; (2) predicts yearly probabilities of physician's choice of province of residence; (3) examines marginal effect of expected income in a province on response (choice of province of residence) probability for physicians residing in each province. We estimated McFadden's conditional logit discrete choice model with yearly probability of choosing a province of residence as a dependent variable for physicians residing in each province separately. The results show: (1) the effect of expected income in a province on the choice of province of residence is positive and statistically significant for physicians residing in Ontario and Saskatchewan. There are other provincial characteristics besides income that affect physician's choice of province of residence; (2) most physicians choose the province they are currently residing. However, those who choose a province other than the current province of residence, the preference for a certain province varies across physicians residing in different provinces; (3) the marginal effect of expected income in a province on response probability varies across provinces for physicians residing in the same province. It also varies across physicians residing in different provinces for the same amount of change in income in the same province. PMID- 16473438 TI - Economic strain and self-rated health among lone and couple mothers in Sweden during the 1990s compared to the 1980s. AB - OBJECTIVES: Changes on the labour market and in Swedish welfare policy during the 1990s may have affected lone mothers. This study analysed economic strain and self-rated health (SRH) among lone and couple mothers in Sweden in the 1980s and the 1990s. PARTICIPANTS: 22,308, mothers, 19,122 couple and 3186 lone mothers, who responded to the Swedish Survey of Living Conditions in the years 1979-1998. METHODS: Exposure for economic strain was defined as having had difficulties to make ends meet in the last year, the outcome measure was less than good SRH. Prevalence rates were calculated and logistic regression analysis was used in the analysis. Adjustments were made for type of mother, age, time period, socio economic group, income, born in Sweden/foreign born and employment. RESULTS: Prevalence rates of economic strain and less than good SRH increased during the 1990s compared to the 1980s among lone and couple mothers. A polarisation in SRH was noted among lone mothers, with worsening health in poorer groups and improved health among better off groups. Economic strain had a substantial explanatory value for the excess risk of less than good SRH throughout the period studied. The association between economic strain and SRH did not change between lone and couple mothers between the 1980s and the 1990s or in different income groups. CONCLUSIONS: The increased prevalence of less than good SRH among sub-groups of lone mothers may in part be due to an increase of financial problems among these groups in Sweden. Economic strain was an important explanatory factor. PMID- 16473439 TI - Excitotoxic mechanisms and the role of astrocytic glutamate transporters in traumatic brain injury. AB - Glutamate excitotoxicity plays an important role in the development of secondary injuries that occur following traumatic brain injury (TBI), and contributes significantly to expansion of the total volume of injury. Acute increases in extracellular glutamate levels have been detected in both experimental brain trauma models and in human patients, and can lead to over-stimulation of glutamate receptors, resulting in a cascade of excitotoxic-related mechanisms culminating in neuronal damage. These elevated levels of glutamate can be effectively controlled by the astrocytic glutamate transporters GLAST (EAAT1) and GLT-1 (EAAT2). However, evidence indicate these transporters and splice variant are downregulated shortly following the insult, which then precipitates glutamate mediated excitotoxic conditions. Lack of success with glutamate receptor antagonists as a potential source of clinical intervention treatment following TBI has resulted in the necessity for a better understanding of the mechanisms that underlie the process of excitotoxicity, including the function and regulation of glutamate transporters. Such new insight should improve the likelihood of development of novel avenues for therapeutic intervention following TBI. PMID- 16473440 TI - Neuropsychological functioning in people with ADHD across the lifespan. AB - ADHD is defined by behavioral characteristics similar to neuropsychological disorders of executive dysfunction. This paper is a literature review of the neurocognitive characteristics of ADHD from early childhood through adulthood. The author addresses the development of the concept of attention and executive function (EF) deficits in ADHD, clinical neuropsychological studies of pre teenage children, teenagers and adults with ADHD, gender and the role of psychiatric co-morbidity including the relationship of learning disabilities to ADHD, heterogeneity of neuropsychological dysfunctions, experimental neuropsychological studies, the relationship of brain structure to function, psychopharmacology of ADHD, and clinical neuropsychological assessment. The group data clearly supports the hypothesis that executive dysfunctions are correlates of ADHD regardless of gender and age, and these EF deficits are exacerbated by co morbidity with learning disabilities such as dyslexia. However, there is limited data on children under the age of 5, teenagers from age 13-18, and adults with ADHD over the age of 40. Studies of individual classification of people with ADHD compared to healthy, non-psychiatric controls do not support the use of neuropsychological tests for the clinical diagnosis of ADHD, and indicate that not all persons with ADHD have EF deficits. Some persons with ADHD may have deficits in brain reward systems that are relatively independent of EF impairments. Future research should clarify the multiple sources of ADHD impairments, continue to refine neuropsychological tools optimized for assessment, and incorporate longitudinal, developmental designs to understand ADHD across the lifespan. PMID- 16473441 TI - Family issues in child anxiety: attachment, family functioning, parental rearing and beliefs. AB - Family studies have found a large overlap between anxiety disorders in family members. In addition to genetic heritability, a range of family factors may also be involved in the intergenerational transmission of anxiety. Evidence for a relationship between family factors and childhood as well as parental anxiety is reviewed. Four groups of family variables are considered: (I) attachment; (II), aspects of family functioning, such as marital conflict, co-parenting, functioning of the family as a whole, and sibling relationships; (III) parental rearing strategies; and (IV) beliefs that parents hold about their child. The reviewed literature provides evidence for an association between each of these family factors and child anxiety. However, there is little evidence as yet that identified family factors are specific to child anxiety, rather than to child psychopathology in general. Moreover, evidence for a relationship between child anxiety and family factors is predominantly cross-sectional. Therefore, whether the identified family factors cause childhood anxiety still needs to be investigated. Further research that investigates mechanisms mediating the relationship between family factors and child anxiety is also called for. Finally, parental beliefs are identified as important predictors of parental behaviour that have largely not been investigated in relation to child anxiety disorders. PMID- 16473442 TI - Growth following adversity: theoretical perspectives and implications for clinical practice. AB - A number of literatures and philosophies throughout human history have conveyed the idea that there is personal gain to be found in suffering, and it is an idea central to the existential-humanistic tradition of psychology. However, it is only relatively recently that the topic of growth following adversity has become the focus for empirical and theoretical work. In this paper, we review theoretical models of growth, and discuss the implications of growth for clinical practice. Three main theoretical perspectives are reviewed, the functional descriptive model, the meta-theoretical person-centered perspective, and the biopsychosocial-evolutionary view. It is proposed that these three approaches to theory offer different but complementary levels of analysis, and that theoretical integration between them is possible. We then go on to explore the implications of this theoretical integration for clinical practice, and conclude with a consideration of the role of therapy in facilitating growth following adversity. PMID- 16473443 TI - Depression research and treatment: are we skating to where the puck is going to be? AB - This paper critically reviews empirical findings regarding current key assumptions underlying the nature and treatment of depression which heavily rely on the DSM approach. This review shows that empirical evidence provides little support for these assumptions. In response to these findings, an etiologically based, biopsychosocial, dynamic interactionism model of depression is proposed. This model could foster further integration in research on depression and assist in the development of guidelines for the treatment of depression that are better informed by research findings and more congruent with complex clinical realities. PMID- 16473444 TI - Biomonitoring: is body burden relevant to public health? AB - Biomonitoring is the study of the presence and concentration of chemicals in humans usually by the measurement of blood, urine or breath (exhaled air). Properly conducted, these data provide a picture of the amount of a chemical or agent actually absorbed into the body for a specific period of time. This review provides a history of biomonitoring, as well as the limitations and potential benefits of these studies. Examples of the proper and possibly improper use of biomonitoring and the impact made on our society are provided. Reasons for having comprehensive national biomonitoring programs are summarized, along with the societal benefits and risks. A brief discussion of the history of the NHANES program and select results from the 2005 Report are presented. By 2010, it has been predicted that the Centers for Disease Control (CDC) will be monitoring nearly 1000 chemicals in persons from all regions of the nation. The measurement of chemicals and biomarkers has revolutionized the field of exposure assessment. Overall, we recommend an approach of careful interpretation, understanding that the data obtained are useful for establishing baseline information about exposure, rather than equating detection with risk. We present suggestions for contextualizing biomonitoring results in order to provide the public with the tools to distinguish genuine health risks from trivial ones. PMID- 16473445 TI - Personal meanings in the construction of need for total knee replacement surgery. AB - This study explores patients' personal meanings of knee osteoarthritis (OA) and total knee replacement (TKR). Personal meanings are important because decisions regarding the need for TKR do not seem to be explained by symptoms alone. A total of 18 semi-structured interviews were conducted with a purposive sample of respondents who were listed for TKR at one UK specialist orthopaedic hospital, and who had lower than average disease burden according to standard quantitative criteria. Data were analysed using interpretive phenomenological analysis. Several themes related to need for TKR emerged: adoption of the medical model, a person's social network, pain, functional loss, feelings of vulnerability, dependency, low mood and fatigue, ideas related to disease progression and expectations of TKR. Results suggest that the decision to undergo TKR is not related to symptoms alone, but to personal meanings. Some of these personal meanings may not be useful in accurately assessing the need for TKR, and may result in mis-targeting of treatment. It is important for health professionals to explore and tackle personal meanings when considering treatment. In particular, it may be important to consider gender differences that may influence when men and women are listed for TKR. For example: women may be less likely to discuss treatment options with their doctor; they may be more likely to have heard negative examples of TKR from friends or family; they may be more likely to discuss pain and mobility issues than activities requiring higher function; and they may be more likely to discuss the effect on their mood and the emotional impact of OA. PMID- 16473446 TI - The "Americanisation" of migrants: evidence for the contribution of ethnicity, social deprivation, lifestyle and life-course processes to the mid-20th century Coronary Heart Disease epidemic in the US. AB - We investigated the contribution of the large-scale immigration of White Europeans into the US between 1850 and 1930 to the timing and extent of the epidemic pattern of heart disease between 1900 and 1980. The analyses are based on data collected through the United States Federal Census from 1850 to the present. The hardcopy historical record confirms that census reports themselves and related monographs were concerned from 1850 with excessive mortality from heart disease of immigrants, particularly of Northern European origin and initially at least, their first-generation native-born children. Our analysis of the electronic database indicates a strong relationship between the percentage of US population foreign born and native born of foreign parentage and age adjusted mortality from heart disease. We identified a lag of 50 years giving the maximum linear correlation coefficient for men (r(2) = 0.92), and for women a shorter lag of 38 years and an earlier decline in Coronary Heart Disease (CHD) rates (r(2) = 0.96). Both the rise and fall of the CHD epidemic over an 80-year period correspond closely to the rise and fall of the foreign population in previous years. For the foreign born only, age adjusted negative binomial general estimated equation (GEE) models calculate the relative risk of dying of heart disease per 10% increase in proportion foreign born. There is an independent influence for men until 1930 and for women throughout the period from 1910 onwards. We conclude there is an impact of immigration on the pattern of the epidemic, mediated through a combination of factors, such as accumulated life course susceptibility, deprived socio-economic conditions upon arrival, and the enthusiastic uptake of behaviours related to the classic risk factors of smoking, high saturated fat and salt diet. Our analysis provides a more contextualised understanding of the scale and timing of the epidemic of CHD in the US. PMID- 16473447 TI - Suicide mortality of Eastern European regions before and after the Communist period. AB - The aim of this study was to investigate the spatial distribution of Eastern European suicide mortality both before and at the end of the Communist period, as well as the changes that occurred during this period. Regional data on suicide mortality were collected from Czarist "European Russia" in 1910 and from the corresponding area in 1989. The distribution of suicide mortality was mapped at both points in time. Regional continuity over time was further studied with the help of geographical units specially constructed for this purpose. In 1910, suicide mortality was found to be high in the northern Baltic provinces, in the urban parts of north and central Russia, the more urbanized parts of northern and western Poland, in east Ukraine, and in the northern Caucasus, while suicide rates were generally low in south Russia, Dagestan, and in southern Poland. In 1989, suicide mortality was highest in the Urals, the east Russian "ethnic" areas, and in southeast Russia. The rates were low in Poland, Moldavia, and in most of the northern Caucasus. The across-time analysis using specially constructed comparison units showed that the spatial distributions of suicide mortality in 1910 and 1989 were not correlated with each other. Additional analyses pointed to a short-term consistency of regional patterns both in the 1900s-1920s and the 1980s-1990s. The lack of regional continuity in suicide mortality in the area may imply an absence of strong and continuous regional cultures, or a strong influence of other factors, such as societal modernization, on suicide mortality. Suicide as an act changed its social nature during the Communist period, becoming more normal, and more equally distributed among social classes and geographical locations. PMID- 16473448 TI - Adolescent suicide attempters in Hong Kong and the United States. AB - Using a nested case-control design drawing from a larger sample of 2272 adolescents, this study examined the characteristics of adolescents from Hong Kong (n = 99) and the United States (US) (n = 60), who indicated that they had made a suicide attempt at least once in the past, compared to their equally depressed sex- and age-matched counterparts who had not attempted suicide. The cross-cultural literature suggested three hypotheses which were tested: (a) depressive symptoms would differentiate between suicide attempters compared to non-attempters less in Hong Kong and more in the US; (b) discord in interpersonal relationships would be a more important distinguishing factor in suicide attempts in Hong Kong than in the US and (c) cognitive variables such as hopelessness would be less associated with suicide attempts in Hong Kong compared to the US. There were strong similarities in the two cultures with regard to characteristics that distinguished between attempters and matched controls. Hypotheses regarding cultural differences between attempters in Hong Kong and the US were not supported. Depression, current and lifetime suicide ideation, hopelessness, poor interpersonal relationships and exposure to suicide attempters and completers distinguished attempters from controls equally in the two cultures and in both genders. These findings do not support the proposition that depression is a less relevant variable in suicide attempts in Chinese versus western groups. PMID- 16473449 TI - Brain activation during semantic processing in autism spectrum disorders via functional magnetic resonance imaging. AB - Language and communication deficits are core features of autism spectrum disorders (ASD), even in high-functioning adults with ASD. This study investigated brain activation patterns using functional magnetic resonance imaging in right-handed adult males with ASD and a control group, matched on age, handedness, and verbal IQ. Semantic processing in the controls produced robust activation in Broca's area (left inferior frontal gyrus) and in superior medial frontal gyrus and right cerebellum. The ASD group had substantially reduced Broca's activation, but increased left temporal (Wernicke's) activation. Furthermore, the ASD group showed diminished activation differences between concrete and abstract words, consistent with behavioral studies. The current study suggests Broca's area is a region of abnormal neurodevelopment in ASD, which may be linked with semantic and related language deficits frequently observed in ASD. PMID- 16473450 TI - Hibiscus anthocyanins-rich extract inhibited LDL oxidation and oxLDL-mediated macrophages apoptosis. AB - The oxidative modification of low-density lipoprotein (LDL) plays a key role in the pathogenesis of atherosclerosis. Anti-oxidative reagents, which can effectively inhibit LDL oxidation, may prevent atherosclerosis via reducing early atherogenesis, and slowing down the progression to advance stages. As shown in previous studies Hibiscus sabdariffa L. is a natural plant containing a lot of pigments that was found to possess anti-oxidative of activity. Therefore, in this study, we evaluated the anti-oxidative activity of Hibiscus anthocyanins (HAs) by measuring their effects on LDL oxidation (in cell-free system) and anti-apoptotic abilities (in RAW264.7 cells). HAs have been tested in vitro examining their relative electrophoretic mobility (REM), Apo B fragmentation, thiobarbituric acid relative substances (TBARS) and radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity assay. The anti-oxidative activity of HAs was defined by relative electrophoretic mobility of oxLDL (decrease of 50% at 2 mg/ml), fragmentation of Apo B (inhibition of 61% at 1mg/ml), and TBARS assay (IC(50): 0.46 mg/ml) in the Cu(2+)-mediated oxidize LDL. Furthermore, the addition of >0.1 mg/ml of HAs could scavenge over 95% of free DPPH radicals, HAs showed strong potential in inhibiting LDL oxidation induced by copper. In addition, to determine whether oxLDL-induced apoptosis in macrophages is inhibited by HAs, we studied the viability, morphology and caspase-3 expression of RAW 264.7 cells. MTT assay, Leukostate staining analysis and Western blotting reveals that HAs could inhibit oxLDL-induced apoptosis. According to these findings, we suggest that HAs may be used to inhibit LDL oxidation and oxLDL-mediated macrophage apoptosis, serving as a chemopreventive agent. However, further investigations into the specificity and mechanism(s) of HAs are needed. PMID- 16473451 TI - Roles of endogenous ascorbate and glutathione in the cellular reduction and cytotoxicity of sulfamethoxazole-nitroso. AB - Sulfamethoxazole (SMX) is an effective drug for the management of opportunistic infections, but its use is limited by hypersensitivity reactions, particularly in HIV-infected patients. The oxidative metabolite SMX-nitroso (SMX-NO), is thought to be a proximate mediator of SMX hypersensitivity, and can be reduced in vitro by ascorbate or glutathione. Leukocytes from patients with SMX hypersensitivity show enhanced cytotoxicity from SMX metabolites in vitro; this finding has been attributed to a possible "detoxification defect" in some individuals. The purpose of this study was to determine whether variability in endogenous ascorbate or glutathione could be associated with individual differences in SMX-NO cytotoxicity. Thirty HIV-positive patients and 23 healthy control subjects were studied. Both antioxidants were significantly correlated with the reduction of SMX-NO to its hydroxylamine, SMX-HA, by mononuclear leukocytes, and both were linearly depleted during reduction. Controlled ascorbate supplementation in three healthy subjects increased leukocyte ascorbate with no change in glutathione, and significantly enhanced SMX-NO reduction. Ascorbate supplementation also decreased SMX-NO cytotoxicity compared to pre-supplementation values. Rapid reduction of SMX-NO to SMX-HA was associated with enhanced direct cytotoxicity from SMX-NO. When forward oxidation of SMX-HA back to SMX-NO was driven by the superoxide dismutase mimetic, Tempol, SMX-NO cytotoxicity was increased, without enhancement of adduct formation. This suggests that SMX-NO cytotoxicity may be mediated, at least in part, by redox cycling between SMX-HA and SMX-NO. Overall, these data indicate that endogenous ascorbate and glutathione are important for the intracellular reduction of SMX-NO, a proposed mediator of SMX hypersensitivity, and that redox cycling of SMX-HA to SMX-NO may contribute to the cytotoxicity of these metabolites in vitro. PMID- 16473452 TI - Etiology of bromate-induced cancer and possible modes of action-studies in Japan. AB - Renal cell tumors were significantly increased in male and female rats given potassium bromate at 250 and 500 mg/L in drinking water. In at least one other study renal cell tumors were produced in male rats at 125 mg/L. Among male mice given 750 mg/L of potassium bromate, there were no significant differences in renal cell tumors between treated and control groups after 88 weeks on test. In oxidative DNA damage tests 8-oxodeoxyguanosine (8-oxodG also referred to as 8-OH dG) was induced in DNA in the male rat kidney in 1 week, and in females after 3 weeks at 500 mg/L, and also in both male and female rats at 250 mg/L, but not at 125 mg/L. DNA adducts are considered to be an initial step in the carcinogenesis process, however, the administered doses are not always sufficient to cause mutations, possibly due to DNA repair. In the two-step rat renal carcinogenesis model using N-ethyl-N-hydroxyethylnitrosamine (EHEN) as initiator, promotion activity by potassium bromate was measured using the BrdU labeling index. The promoting activity of bromate in male rats was much greater and extended to doses as low as 60 mg/L in male rats, whereas in females the response was limited to 250 and 500 mg/L. Therefore, it was concluded that the mechanisms contributing to cancer in the male rat were more complex than in the female rat. The accumulation of alpha2mu-globulin in the kidneys of male rats exposed to potassium bromate probably accounts for the greater labeling index in the male rat relative to the female rat. Accumulation of alpha(2mu)-globulin as a result of treatment with chemicals is unique to the male rat and does contribute to carcinogenic responses. Neither humans nor female rats display this response. Nevertheless, bromate must be considered carcinogenic because of the response of the female rats. The better correlation between 8-oxodG formation and tumor response indicates that dose-response information from the female rat would be much more relevant to human risk assessment. The fact that an elevation of BrdU-LI in the kidney of the female rat is consistent with the possibility that cell proliferation observed in female rats resulted from oxidative stress and/or cytotoxic responses in the kidney. Therefore, oxidative stress is most likely the mechanism of interest for cancer risk in humans. PMID- 16473453 TI - Enzyme induction in cryopreserved human hepatocyte cultures. AB - Freshly isolated human hepatocytes are considered as the gold standard for in vitro testing of drug candidates. Meanwhile also cryopreserved human hepatocyte suspensions are available. However, a drawback of these cells is the incalculability of attachment to the culture dish. Therefore, we established a technique freezing hepatocytes cultured on a collagen gel. After thawing damaged cells were removed to a certain extent by gentle washing with culture medium prior to adding an upper gel layer. The morphology of the resulting hepatocyte cultures could not be distinguished from that of non-frozen cells. However, basal activities of cytochrome P450 isoforms decreased in cryopreserved compared to non frozen hepatocytes, as evidenced by analysis of testosterone hydroxylation (OHT) in positions 6beta, 16alpha, 2beta and 6alpha. Nevertheless, enzyme induction factors caused by 24 h incubation with 50 microM rifampicin were similar in cryopreserved and non-frozen hepatocytes. In cryopreserved hepatocytes rifampicin caused an increase in mean values of 6beta-OHT formation from 57.2 to 157.7 pmol/well/min (2.8-fold), compared to an increase from 115.8 to 269.1 pmol/well/min (2.3-fold) in non-frozen cells. Similarly, 16alpha- and 2beta-OHT showed induction factors of 2.4- and 2.3-fold in cryopreserved compared to 1.6- and 2.4-fold in non-frozen hepatocytes, respectively. In conclusion, human hepatocytes cryopreserved on collagen gels show a clear induction of CYP3A4 by rifampicin, although the basal activities are reduced compared to non-frozen cells. PMID- 16473454 TI - A preliminary study on the relationship between stimulated saliva and periodontal conditions in community-dwelling elderly people. AB - OBJECTIVES: The purpose of this study was to explore the relationship between flow rate and spinnbarkeit of stimulated whole saliva and periodontal conditions in healthy elderly people. METHODS: Three hundred and fifty-five dentate subjects aged 76 years were included. The pocket probing depth (PD), attachment level (AL), and bleeding on probing (BOP) were measured. Stimulated whole saliva was collected and the salivary flow rate (SFR) was calculated. Then, salivary spinnbarkeit (SS) was immediately measured. RESULTS: The mean SFR and SS were 1.44 ml/min and 1.91 mm, respectively. SFR was not significantly related to each periodontal parameter. On the other hand, subjects with SS2.00 mm exhibited a significantly higher mean PD (p<0.05), % of sites with PD>or=4mm (p<0.05), mean AL (p<0.01), % of site with AL>or=4mm (p<0.05) and % of sites with AL>or=6mm (p<0.01) than subjects in the other three groups (one-way ANOVA). In the logistic regression analysis, the factors significantly associated with the highest quintile of PD>or=4mm were: the low salivary flow and the high salivary spinnbarkeit (OR 3.84), current smokers (OR 5.08), cleaning interdental spaces rarely/never (OR 2.12), and frequent BOP (OR 5.20). CONCLUSION: These findings suggest that high salivary spinnbarkeit in addition to a low salivary flow rate might be a high risk for periodontal disease in elderly people. PMID- 16473455 TI - Mnemonic functions of the hippocampus: a comparison between animals and humans. AB - This review summarizes a series of experiments aimed at answering the question whether the hippocampus in rats and humans performs parallel functions focusing on studies that assess spatial and temporal pattern separation, sequential learning, spatial and temporal pattern association, spatial and temporal pattern completion, and short-term and intermediate-term memory. It is recognized that a comparison of the functions of the rat hippocampus with human hippocampus is difficult, because of differences in methodology, differences in complexity of life experiences, and differences in the degree of hippocampal damage as well as damage to interconnected brain regions. Yet, in general the data support the idea that with respect to spatial and temporal pattern separation, sequential learning, spatial and temporal pattern associations, spatial and temporal pattern completion, and short-term and intermediate-term memory, similar functions are observed in rats and humans with hippocampal damage using analogous tasks. These data provide support for evolutionary continuity in cognitive function assigned to the hippocampus of rats and humans. PMID- 16473456 TI - Sub-typing CFS patients on the basis of 'minor' symptoms. AB - The diagnosis of chronic fatigue syndrome (CFS), an illness characterized by medically unexplained fatigue, depends on a clinical case definition representing one or more pathophysiological mechanisms. To prepare for studies of these mechanisms, this study sought to identify subtypes of CFS. In 161 women meeting 1994 criteria for CFS, principal components analysis of the 10 'minor' symptoms of CFS produced three factors interpreted to indicate musculoskeletal, infectious and neurological subtypes. Extreme scores on one or more of these factors characterized about 2/3 of the sample. Those characterized by the neurological factor were at increased risk of reduced scores on cognitive tests requiring attention, working memory, long-term memory or rapid performance. In addition, the neurological subtype was associated with reduced levels of function. Those characterized by the musculoskeletal factor were at increased risk for the diagnosis of fibromyalgia (chronic widespread pain and mechanical allodynia) and reduced physical function. Those characterized by the infectious factor were less likely to evidence co-occurring fibromyalgia, and showed lesser risk of functional impairment. The prevalence of disability was increased in those with the highest scores on any of the subtypes, as well as in those with high scores on multiple factors. Depression and anxiety, while frequently present, were not more prevalent in any particular subtype, and did not increase with the severity of specific symptom reports. Results suggest that subtypes of CFS may be identified from reports of the minor diagnostic symptoms, and that these subtypes demonstrate construct validity. PMID- 16473457 TI - Interfacial properties of the M1 segment of the nicotinic acetylcholine receptor. AB - We have studied the thermodynamic, surface, and structural properties of alphaM1 transmembrane sequence of the nicotinic acetylcholine receptor (nAChR) by using Langmuir monolayer, FT-IR spectroscopy and molecular dynamics simulation techniques in membrane-mimicking environments. M1 spontaneously incorporates into a lipid-free air-water interface, showing a favourable adsorption free energy of 7.2 kcal/mol. A cross-sectional molecular area of 210 A(2)/molecule, a surface potential of 4.2 fV/molecule and a high stability of the film were deducted from pure M1 monolayers. FT-IR experiments and molecular dynamics simulations in membrane-mimicking environments (sodium-dodecyl-sulfate and CCl(4), respectively) indicate coexistence between helical and non-helical structures. Furthermore, mixed peptide-lipid monolayers and monolayer penetration experiments were performed in order to study the peptide-lipid interaction. Mixed with condensed lipids (dipalmitoyl-phosphocholine, and dipalmitoyl-phosphoglycerol), M1 shows immiscible/miscible behaviour at low/high peptide concentration, respectively. Conversely, a complete miscible peptide-lipid interface is observed with liquid expanded lipids (palmitoyl-oleoyl-phosphocholine, and palmitoyl-oleoyl phosphoglycerol). Peptide penetration experiments demonstrate that the M1 peptide preferentially interacts with zwitterionic phosphocholine interfaces. PMID- 16473458 TI - Penile suspensory ligament division for penile augmentation: indications and results. AB - OBJECTIVE: This study assessed the management of patients requesting penile length enhancement by division of the penile suspensory ligament. METHODS: From September 1998 to January 2005, 42 patients with a variety of etiologies were included; all underwent division of the penile suspensory ligament. The outcome was assessed objectively based on increase in flaccid stretched penile length (SPL) and subjectively using the rates of patient satisfaction. RESULTS: The mean increase in SPL was 1.3+/-0.9 cm (range, -1 to +3 cm), with the addition of a silicone spacer placed between the pubis and penis giving a better outcome (p<0.05). The overall patient satisfaction rate was 35% but lower in the group with penile dysmorphic disorder at 27%. CONCLUSION: Division of the penile suspensory ligament or other augmentation techniques may increase penile length but usually not to a degree that satisfies the patient. Men with penile dysmorphic disorder often have unrealistic expectations regarding the outcome of surgical intervention and should be encouraged to seek psychological help primarily, with surgery reserved as the last resort. PMID- 16473459 TI - Developmental expression of Manduca shade, the P450 mediating the final step in molting hormone synthesis. AB - The ecdysone 20-monooxygenase (E20MO; 20-hydroxylase) is the enzyme that mediates the conversion of ecdysone (E) to the active insect molting hormone, 20 hydroxyecdysone (20E), which coordinates developmental progression. We report the identification and developmental expression of the Halloween gene shade (shd; CYP314A1) that encodes the E20MO in the tobacco hornworm, Manduca sexta. Manduca Shd (MsShd) mediates the conversion of E to 20E when expressed in Drosophila S2 cells. In accord with the central dogma, the data show that Msshd is expressed mainly in the midgut, Malpighian tubules, fat body and epidermis with very low expression in the prothoracic gland and nervous system. Developmental variations in E20MO enzymatic activity are almost perfectly correlated with comparable changes in the gene expression of Msshd in the fat body and midgut during the fifth instar and the beginning of pupal-adult development. The results indicate three successive and overlapping peaks of expression in the fat body, midgut and Malpighian tubules, respectively, during the fifth larval instar. The data suggest that precise tissue-specific transcriptional regulation controls the levels, and thereby the activity, of the Manduca E20MO. PMID- 16473460 TI - Stimulation of the motor cortex for disabling essential tremor. AB - OBJECTIVE: To examine the safety and efficacy of targeted stimulation of the motor cortex as a treatment for essential tremor (ET). PATIENTS AND METHODS: At the University of Kansas Medical Center, two patients with essential tremor received stimulation of the (contralateral) motor cortex using an investigational implantable pulse generator (IPG). Patients were evaluated with the Fahn Tolosa Marin tremor rating scale (TRS) at baseline, 1 week and 4 weeks after surgery, both with stimulation turned on and turned off. Both patients also received neuropsychological assessments at baseline and again after surgery. RESULTS: Patient 1 was a 75-year-old male with tremor for 20 years. His baseline total TRS score was 61 and his TRS 1 month after surgery was 57. His IPG was set at 30 Hz, 3 mA and 250 micros pulse width. Patient 2 was a 60-year-old male with tremor for over 10 years. His baseline total TRS was 47 and it was 43, 1 month after surgery. His IPG was set at 50 Hz, 5 mA and 250 micros pulse width. There were no adverse effects. CONCLUSION: Cortical stimulation of the primary hand motor cortex contralateral to the dominant hand was ineffective for the treatment of ET with the stimulation parameters used in this study. Future research examining other stimulation parameters is necessary to determine if there is a role for cortical stimulation in the treatment of ET. PMID- 16473461 TI - Urinary cytokeratin 20 mRNA expression has the potential to predict recurrence in superficial transitional cell carcinoma of the bladder. AB - Higher levels of cytokeratin 20 (CK 20) mRNA are expressed in malignant urothelial tissue compared to normal tissue. We determined the CK 20 mRNA expression in urine from patients with transitional cell carcinoma (TCC) of the bladder and assessed the biological behavior of such tumors in a 5-year follow up. Second voided urine was preoperatively collected from 56 patients with bladder carcinoma, from 20 patients with nonmalignant urological diseases and from 40 healthy volunteers. RNA extraction from exfoliated urothelial cells was followed by quantitative real-time RT-PCR with the Light Cycler. Patients in the superficial TCC group had a median expression of 8226AU (arbitrary units) with and 1523AU without tumor recurrence (P=0.023). No such correlation was detected in the group with muscle-invasive tumors. Kaplan-Meier analysis revealed a significant difference between recurrent and nonrecurrent disease (P=0.019) in superficial but not in muscle-invasive TCC (P=0.84). CK 20 mRNA expression in urine has the potential to identify patients at risk for recurrence of noninvasive papillary urothelial tumors. It helps to categorize patients prior to TUR-B, so that the cystoscopy interval during follow-up may be extended in those with low-risk superficial TCC. PMID- 16473463 TI - Potentiation of morphine antinociception by pentobarbital in female vs. male rats. AB - It has been shown previously that female rats are more sensitive than males to barbiturate anesthesia, whereas males may be more sensitive than females to opioid antinociception. The aim of the present study was to determine whether enhancement of morphine antinociception by pentobarbital, previously demonstrated in male animals and humans, occurs similarly in females. Pentobarbital (50 mg/kg i.p.) produced longer-lasting anesthetic effects (loss of muscle tone, righting reflex) in gonadally intact female rats than in males, but greater antinociceptive effects in males at some time points post-injection. There were no significant sex differences in morphine-induced anesthesia or antinociception; however, 50 mg/kg pentobarbital produced greater leftward shifts in the morphine antinociceptive dose-effect curve in gonadally intact females than males, whether pentobarbital was administered 30 vs. 120 min before morphine (times at which there were no sex differences vs. sex differences, respectively, in pentobarbital's effects when administered alone). Dose-addition analysis confirmed that pentobarbital enhancement of morphine antinociception was supra additive in both sexes; morphine also significantly enhanced pentobarbital induced anesthesia in both sexes. In gonadectomized males, testosterone did not significantly alter pentobarbital enhancement of morphine antinociception; in contrast, in gonadectomized females, estradiol significantly attenuated the drug interaction. Estradiol did not significantly alter the effects of pentobarbital alone or morphine alone, indicating that the attenuation of the pentobarbital's potentiation of morphine antinociception in estradiol-treated rats is specific to the drug interaction. These results suggest that barbiturate potentiation of opioid antinociception may be greater in females - particularly those in low ovarian hormone states - than in males. PMID- 16473462 TI - Loss of expectation-related mechanisms in Alzheimer's disease makes analgesic therapies less effective. AB - Expectation/placebo-related mechanisms and specific effects of therapies show additive effects, such that a therapy is less effective if the placebo component is absent. So far, the placebo component has been disrupted experimentally by using covert administrations of treatments. Here, we show for the first time that disruption of expectation/placebo-related analgesic mechanisms may occur in a clinical condition, Alzheimer's disease (AD). In order to assess the placebo component of a therapy, we used the recently developed open-hidden paradigm. A local anesthetic was applied, either overtly or covertly, to the skin of AD patients to reduce burning pain after venipuncture. The placebo (psychological) component is represented by the difference between the analgesic effect after open (expected) and after hidden (unexpected) application. We correlated the placebo component with both cognitive status and functional connectivity among different brain regions. We found that AD patients with reduced Frontal Assessment Battery scores showed reduced placebo component of the analgesic treatment. We also found that the disruption of the placebo component occurred when reduced connectivity of the prefrontal lobes with the rest of the brain was present. Remarkably, the loss of these placebo-related mechanisms reduced treatment efficacy, such that a dose increase was necessary to produce adequate analgesia. These findings highlight the active role of cognition and prefrontal lobes in the therapeutic outcome and underscore the need of considering a possible revision of the therapeutic approach in Alzheimer patients in order to compensate for the loss of the endogenous expectation and placebo mechanisms. PMID- 16473464 TI - Recurrent pain symptoms in young schoolchildren are often multiple. AB - Different recurrent pain symptoms in children have mainly been investigated separately and comprehensive studies exploring the relationship between several of the most common recurrent pain symptoms simultaneously are few. The present study investigated frequency and co-occurrence of recurrent headache, stomach ache, and backache in young schoolchildren (6-13 years of age). A randomized cluster sample of 1155 children from grades 0 to 6 completed a questionnaire; the youngest children helped by their parent. The results showed that 2/3 of the children reported having pain at least once every month, 1/3 at least once a week, and 6% reported experiencing pain symptoms every day. Half of the children with recurrent pain symptoms reported pain symptoms from several body locations, and, in children with weekly pain symptoms, two out of three reported multiple pain. Multiple, but not single, pain symptoms became more prevalent with age. The only difference between girls and boys was a higher prevalence of multiple weekly pain symptoms in girls. In conclusion, this study shows that a great number of young schoolchildren suffer from frequently recurring and co-occurring pain symptoms, indicating an urgent need for preventive and curative programs starting already in the first school years. Furthermore, the high prevalence of multiple symptoms indicates that recurrent pain symptoms in children, particularly frequent symptoms, should be regarded a potential general pain disorder rather than merely a localized body disorder. PMID- 16473465 TI - Efficacy of an injectable formulation of eprinomectin against Psoroptes cuniculi, the ear mange mite in rabbits. AB - Thirty rabbits naturally infected with ear mange mite, Psoroptes cuniculi, were subcutaneously administrated with a single dose of eprinomectin at 100, 200 and 300 microg/kg body weight or vehicle on day 0. The extent of lesions was scored on day -6 (prior to treatment), day 0 (treated), days 7, 14, 21, 28 and 35, the ear scabs were collected simultaneously; mites in scabs were examined and counted. The results showed that a single dose of eprinomectin at 200 or 300 microg/kg body weight following subcutaneous administration was able to eliminate P. cuniculi infection in rabbits, and a dose of eprinomectin at 100 microg/kg could significantly reduce mites but was unable to eliminate P. cuniculi. PMID- 16473466 TI - Epidemiological study of the cystic echinococcosis in Morocco. AB - The objectives of this epidemiological study on cystic echinococcosis (CE) in Morocco (2001-2004) were to update the prevalence of CE in different animal species living in the most important areas of the country and to collect protoscoleces and germinal layers for genetic research purposes. The post mortem inspection concerned 2948 sheep, 2337 goats, 618 cattle, 482 camels and 455 equines (325 horses, 60 mules and 70 donkeys) in five different regions: the Rif (Mediterranean coast and high mountains of the Rif), the Loukkos (Atlantic northwest plain), the center (Rabat and Casablanca regions), the Middle Atlas mountains and the south (arid and semi desert areas). The global CE infection prevalence rates obtained were 22.98% in cattle, 10.58% in sheep, 12.03% in camels, 17.80% in equines and 1.88% in goats. The infection rates were especially high in the Middle Atlas in cattle (48.72%) and in the Loukkos in cattle and sheep (37.61 and 31.65%, respectively). The majority of infected cattle (49.6%) and sheep (52.1%) had hydatid cysts in both liver and lungs. Except for cattle, the liver was more infected than lungs in all the other animal species. Animals more than 5 years old were the most infected in all species. The mean CE infection rates of these animals were about 56% in cattle, 40% in sheep, 20% in camels, 17.80% in equines and 7% in goats. These rates were much higher in the Loukkos (85% of cattle and 59% of sheep) and in the Middle Atlas (68% of cattle and 45% of sheep) than in the other regions. Results showed that Echinococcus granulosus is in an endemic steady state with no evidence of protective immunity in the intermediate hosts. The mean numbers of infections per year are 0.099 for cattle, 0.063 for sheep, 0.03 for camels and 0.010 for goats. PMID- 16473467 TI - Advantages of real-time PCR assay for diagnosis and monitoring of canine leishmaniosis. AB - The aim of the present study is to highlight the advantages of real-time quantitative PCR intended to aid in the diagnosis and monitoring of canine leishmaniosis. Diagnosis of canine leishmaniosis is extremely challenging, especially in endemic areas, due to the diverse and non-specific clinical manifestations, and due to the high seroprevalence rate in sub-clinical dogs. Veterinarian clinicians are usually confronted with cases that are compatible with the disease, and with several diagnostic tests, sometimes with contradictory results. We have developed a new TaqMan assay, targeting the kinetoplast, applied to 44 samples of bone marrow aspirate or peripheral blood. The dynamic range of detection of Leishmania DNA was established in 7 logs and the limit of detection is 0.001 parasites in the PCR reaction. At the time of diagnosis parasitemia ranges from less than 1 to 10(7)parasites/ml. The ability to quantify the parasite burden allowed: (i) to elucidate the status of positive dogs by conventional PCR, although larger studies are necessary to clarify the dividing line between infection and disease, (ii) to estimate the kinetics of the parasite load and the different response to the treatment in a follow-up and (iii) to validate blood as less invasive sample for qPCR. The continuous data provided by real-time qPCR could solve the dilemma for the clinician managing cases of canine leishmaniosis by differentiating between Leishmania-infected dogs or dogs with active disease of leishmaniosis. PMID- 16473468 TI - The rabbit coccidium Eimeria piriformis: Selection of a precocious line and life cycle study. AB - Specific pathogen-free (SPF) rabbits were inoculated with oocysts of an original strain (OS) of Eimeria piriformis and the first newly developed oocysts recovered from the intestine were used for infection of other rabbits. The prepatent period (PP) was shortened after 12 passages from 194 to 170 hours and remained stable after 5 passages without any selection pressure. Oocysts of the precocious line (PL) exhibited peculiar morphology. Besides refractile bodies (RB) within sporozoites, one huge RB joined with a residual body was present inside each sporocyst. The parasite developed in the proximal colon and, to a lesser extent, in other parts of the large intestine. All stages were seen in the epithelium of crypts. In OS, four asexual generations preceded gamogony and, like in other rabbit coccidia, two types of meronts were observed: meronts of type A that develop into polynucleate merozoite, in which endomerogony takes place, and meronts of the type B that form uninucleate merozoites. The endogenous development of PL was identical with that of OS except for the last merogony which was absent. This resulted in earlier appearance of gamogony and shortening of PP. These observations of the life cycle of E. piriformis substantially improve on its description made 50-60 years ago. PMID- 16473469 TI - Glycomics investigation into insulin action. AB - Defects in glycosylation are becoming increasingly associated with a range of human diseases. In some cases, the disease is caused by the glycosylation defect, whereas in others, the aberrant glycosylation may be a consequence of the disease. The implementation of highly sensitive and rapid mass spectrometric screening strategies for profiling the glycans present in model biological systems is revealing valuable insights into disease phenotypes. In addition, glycan screening is proving useful in the analysis of knock-out mice where it is possible to assess the role of glycosyltransferases and glycosidases and what function they have at the cellular and whole organism level. In this study, we analysed the effect of insulin on the glycosylation of 3T3-L1 cells and the effect of insulin resistance on glycosylation in a mouse model. Transcription profiling of 3T3-L1 cells treated with and without insulin revealed expression changes of several glycogenes. In contrast, mass spectrometric screening analysis of the glycans from these cells revealed very similar profiles suggesting that any changes in glycosylation were most likely on specific proteins rather than a global phenomenon. A fat-fed versus carbohydrate-fed mouse insulin resistant model was analysed to test the consequences of chronic insulin resistance. Muscle and liver N-glycosylation profiles from these mice are reported. PMID- 16473470 TI - Vinorelbine in the management of breast cancer: New perspectives, revived role in the era of targeted therapy. AB - Vinorelbine is a semi-synthetic vinca alkaloid that has been shown active in many tumour types and is currently registered for the treatment of advanced breast cancer (ABC) and non-small cell lung cancer (NSCLC). This agent has a generally favourable safety profile, and may be suitable for use in special populations such as the elderly and/or frail patient. However, with the taxanes firmly established as standard second line treatment for ABC after failure of an anthracycline, vinorelbine has been generally relegated for use as third line therapy, in competition with the oral compound capecitabine. More recently, the exciting results observed with the combination of vinorelbine and trastuzumab in patients with Her-2 overexpressing/amplified tumours, as well as the development of a reliable formulation and revised schedule of oral vinorelbine with proven activity in ABC appear to have revived the interest in this compound in the management of this disease. There are still a number of unanswered questions that will have to be addressed by properly designed, adequately powered randomised clinical trials. PMID- 16473471 TI - Long splice variant N type calcium channels are clustered at presynaptic transmitter release sites without modular adaptor proteins. AB - The presynaptic N type Ca channel (CaV2.2) is associated with the transmitter release site apparatus and plays a critical role in the gating of transmitter release. It has been suggested that a distinct CaV2.2 long C terminal splice variant is targeted to the nerve terminal and is anchored at the release face by calcium/calmodulin-dependent serine protein kinase (CASK) and Munc-18-interacting protein (MINT), two modular adaptor proteins. We used the isolated chick ciliary ganglion calyx terminal together with two new antibodies (L4569, L4570) selective for CaV2.2 long C terminal splice variant to test these hypotheses. CaV2.2 long C terminal splice variant was present at the presynaptic transmitter release sites, as identified by Rab3a-interacting molecule (RIM) co-staining and quantitative immunocytochemistry. CASK was also present at the terminal both in conjunction with, and independent of its binding partner, MINT. Immunoprecipitation of CaV2.2 long C terminal splice variant from brain lysate coprecipitated CASK, confirming that these two proteins can form a complex. However, CASK was not colocalized either with CaV2.2 long C terminal splice variant or the transmitter release site marker RIM at the calyx terminal release face. Neither was MINT colocalized with CaV2.2 long C terminal splice variant. Our results show that native CaV2.2 long C terminal splice variant is targeted to the transmitter release sites at an intact presynaptic terminal. However, the lack of enrichment of CASK at the release site combined with the failure of this protein or its partner MINT to colocalize with CaV2.2 argues against the idea that these modular adaptor proteins anchor CaV2.2 at presynaptic nerve terminals. PMID- 16473472 TI - Effect of morphine on the release of excitatory amino acids in the rat hind instep: Pain is modulated by the interaction between the peripheral opioid and glutamate systems. AB - Behavioral evidence supports a role for peripheral glutamate receptors in normal nociceptive transmission. In this study, we examined the release of the excitatory amino acids, glutamate and aspartate, in the s.c. perfusate of the rat hind instep by in vivo microdialysis. Antidromic stimulation of the sciatic nerve and noxious stimuli in the form of heat stimulation and local application of capsaicin cream (1%) to the instep caused an increase in excitatory amino acid release. This capsaicin-induced excitatory amino acid release was suppressed by pretreatment with capsaicin. Both systemic (10 mg/kg, i.v.) and local injections (10(-5) M in the perfusate) of morphine inhibited the increase in excitatory amino acid release evoked by local application of capsaicin cream to the instep. This inhibitory effect of morphine was antagonized by naloxone either given systemically (5 mg/kg, i.v.) or locally (10(-5) M). These results suggest that excitatory amino acids are released from small diameter afferent fibers by heat stimulation in the periphery or local application of capsaicin cream, and that activation of opioid receptors, present on the peripheral endings of small diameter afferent fibers, can regulate noxious stimulus-induced excitatory amino acid release. PMID- 16473473 TI - Enhanced hippocampal GABAergic inhibition in mice overexpressing heparin-binding growth-associated molecule. AB - Heparin-binding growth-associated molecule is a developmentally regulated extracellular matrix protein promoting neurite outgrowth, axonal guidance and synaptogenesis. In the hippocampus, heparin-binding growth-associated molecule is expressed in an activity-dependent manner, and has been shown to suppress long term potentiation of glutamatergic synapses in the area CA1, but the mechanisms underlying this action are unknown. One of the mechanisms by which extracellular matrix proteins might modulate fast synaptic transmission is by altering GABAergic function. Therefore, we have studied the properties of GABAA receptor mediated inhibition in hippocampus of mutant mice overexpressing heparin-binding growth-associated molecule (heparin-binding growth-associated molecule transgenics). Under control conditions the wild-type mice have much higher level of long-term potentiation than the transgenics. However, in the absence of the GABAA receptor-mediated-inhibition a similar level of long-term potentiation is seen in both strains. In field potential recordings blockade of GABAA receptors by picrotoxin resulted in more accentuated increase in the CA1 population spike in the transgenics than in the wild-type animals. Whole-cell patch-clamp recordings revealed that when compared with the wild-type animals the transgenic mice had higher frequency of spontaneous inhibitory postsynaptic currents in CA1 pyramidal neurons. However, the frequency of action potential-independent miniature inhibitory postsynaptic currents was similar in both strains. Further, the transgenics had reduced paired-pulse depression of inhibitory postsynaptic currents, which was insensitive to the blockade of GABAB receptors in contrast to wild-type mice. The results demonstrate that the mice overexpressing heparin binding growth-associated molecule have accentuated hippocampal GABAA receptor mediated inhibition, which in turn may explain the lowered predisposition of glutamatergic synapses to undergo plastic changes in these animals. Thus, our findings suggest a mechanism by which heparin-binding growth-associated molecule can regulate synaptic plasticity. PMID- 16473475 TI - Reported distress associated with concern about a cigarette smoker. AB - It is well documented that concerned others of alcohol abusers report distress associated with the substance user's behavior. No previous study has examined distress associated with concern for someone who smokes cigarettes. To investigate this association, college students, ages 18-24 (N=1719), completed a survey assessing whether or not they were close to someone who smokes whom they thought should quit and the level of distress they experienced regarding this person's smoking. 827 (48.1%) respondents endorsed knowing a smoker whom they thought should quit and 60% reported experiencing at least some distress regarding this person's smoking. From multivariate analysis, the presence of distress (any vs. none) was associated with female gender (OR=1.8, 95% C.I. 1.2, 2.5, p=0.001), never tobacco user (OR=2.0, 95% C.I. 1.4, 2.9, p<0.001), and elevated Perceived Stress Scale score (OR=1.05, 95% C.I. 1.02, 1.08, p<0.001, per 1 unit increase in PSS score). These findings contribute to our understanding of how smoking impacts the psychological well being of another and is intended to stimulate future research investigating the potential for distressed individuals to serve as change agents in tobacco control efforts. PMID- 16473474 TI - Corticosterone inhibits expression of the microglial glutamate transporter GLT-1 in vitro. AB - The present study investigates the effect of the glucocorticoid corticosterone on microglial glutamate transporters in vitro. Microglial cultures obtained from rat cerebral cortex were found to express the excitatory amino acid transporter GLT 1, but not GLAST, and this expression was increased by 1 ng/ml lipopolysaccharide after 12 h of stimulation. This increase has previously been shown to be mediated by tumor necrosis factor-alpha, a cytokine released by microglia during pathological conditions. Furthermore, lipopolysaccharide increased the microglial release of tumor necrosis factor-alpha and 1 microM corticosterone inhibited this effect. Corticosterone also inhibited the lipopolysaccharide-induced increase of the GLT-1 expression as well as the expression in non-activated cells. The effect of corticosterone on the GLT-1 expression was dose dependent and accompanied by similar effects on the microglial glutamate uptake capacity. Additionally, exogenous tumor necrosis factor-alpha was found to counteract the effect of corticosterone on microglial GLT-1 expression. The effect of corticosterone appeared to be glucocorticoid receptor specific since 10 microM of the glucocorticoid receptor antagonist mifepristone inhibited the effect. Thus, corticosterone decreased the microglial uptake of glutamate by decreasing the expression of glutamate transporters, probably due to the inhibited microglial tumor necrosis factor-alpha release. These results provide insights into the mechanisms behind microglial glutamate transporter expression during pathological conditions, and contribute to the debate about the beneficial or harmful effects of glucocorticoids. PMID- 16473476 TI - Cigarette smoking and interest in quitting in methadone maintenance patients. AB - OBJECTIVES: To examine tobacco use, readiness to quit, and interest in smoking cessation interventions among methadone maintenance patients. METHODS: Cross sectional survey of outpatients enrolled in four urban methadone maintenance clinics. Stage of readiness to quit was determined for all smokers, and factors associated with both readiness to quit and interest in attending an on-site smoking cessation program were determined. RESULTS: Among 389 patients, 83% were current smokers. Nearly half (48%) of smokers were contemplating quitting, and an additional 22% were in the preparation stage of readiness to quit. In multivariate analyses, lower nicotine dependence, prior use of smoking cessation pharmacotherapy, and lower methadone dose were associated with being in the preparation stage. Patients with more education, Hispanics/Latinos, and patients who had used smoking cessation pharmacotherapy or were in the preparation or contemplation stages of behavior change were more interested in attending an on site smoking cessation program. CONCLUSIONS: Tobacco use is highly prevalent among methadone maintenance patients, but we also observed a high level of readiness to quit and interest in smoking cessation. Targeted smoking cessation interventions, including on-site programs, should be developed for methadone maintenance patients. PMID- 16473477 TI - The human stratum corneum as extended, covalently cross-linked biopolymer: mathematics, molecules, and medicine. AB - A novel mathematical and molecular hypothesis is proposed to account for the peculiar organization of human epidermis. Mathematically, the organization of the interfollicular epidermis is hypothesized to be a tetratomic identity manifesting a gravitational logic in the arrangement of its functional compartments. The squares of the natural numbers; i.e., 1, 4, 9, and 16 are taken, on empirical grounds, to correspond to the number of cell layers in the respective epidermal strata (germinativum, spinosum, granulosum, and corneum). The outer two strata, overlying the Langerhans cells, constitute the 'living' and 'dead' components of the traditional 'epidermal barrier'. Together, these two strata illustrate in their union of 9 + 16 = 25 cells, a way of conceiving the skin surface (the body environment identity) as both closure and contact. The organization of human epidermis into functional units based on phi, the golden section ratio, builds upon this gravitational logic. Finally, the fact that the extensively cross linked proteolipid envelope of the cornified epidermal cell is a single multi gene molecule is deemed scientifically incontrovertible. The molecular hypothesis in need of validation and verification is whether the corneodesmosomal 'rivets' linking one corneocyte to another are covalently bonded structures. If so, the cornified scaffolding of the stratum corneum constitutes a highly organized, extended, multi-gene, polymer molecule strategically located precisely at the shared surface of the body and environment. This hypothesis places the differentiated structure of the epidermis, an ectodermal derivative like the brain, front and center in the translation of molecular biology to clinical bedside care. PMID- 16473478 TI - [Comparison of cell culture and RT-PCR for the detection of enterovirus in sewage and shellfish]. AB - Enteric viruses contaminating the environment represent a danger for public health notably enteroviruses that are excreted in stools and can contaminate wastewater and shellfish. The ability of enteroviruses to grow in cell culture and the development of techniques of molecular biology applied to their detection make these viruses a good marker of viral pollution of aquatic environment. The aim of our study was to develop a rapid and sensitive RT-PCR technique, able to detect enterovirus RNA in wastewater and shellfish. From 26 samples of wastewater and 56 samples of shellfish, 50.0 and 42.8% were found positive by RT-PCR, respectively, whereas 38.4 and 28.5% were positive by culture, respectively (P=0.077 by chi square test). The two techniques were found concordant in 57.3% of the 82 combined samples, whereas 23 samples (28.0%) were positive only by RT PCR and that 12 samples (14.6%) were positive only by culture. These discrepancies illustrate the fact that the two techniques are not equivalent: the molecular technique allows the detection of not cultivable samples but is sensitive to PCR inhibitors that are present in large amounts in environmental samples. PMID- 16473479 TI - [Investigation of the new QNR-based mechanism of quinolone resistance among enterobacterial strains isolated in Henri-Mondor hospital 2002-2005]. AB - AIM OF THE STUDY: To assess the prevalence of the novel plasmid-mediated resistance to quinolones in enterobacteria isolated in our hospital. MATERIALS AND METHODS: We have screened 737 enterobacterial strains isolated in Henri Mondor hospital between 2002 and 2005 for the presence of the qnr gene by PCR using specific primers. Among them, 282 had a phenotype in concordance with extended spectrum betalactamase (ESBL). Qnr-positive strains were phenotypically and genetically characterized, and epidemiological link between the cases was investigated. RESULTS: Five qnr+ strains were described. The global prevalence was 0.7% but 5/282 among ESBL producing strains and 0/437 among quinolone resistant enterobacteria non producing ESBL. The sequences of the PCR products were identical to qnrA in the environment of the integron In36. All the strains harboured also the ESBL SHV-12 gene. Transfer of qnr by conjugation raised quinolone MICs from 2 to 24 times. However clinical strains harboured a higher level of quinolone resistance and harboured also DNA gyrase and topoisomerase IV mutations. Two strains were epidemiologically related by molecular typing and contact tracing revealed that the patients have been previously hospitalized in the same tertiary care center. CONCLUSION: We described the first investigation of qnr-positive strains in one hospital in France over 4 years. Although the qnr gene prevalence is low, nosocomial transmission is already shown and the transfer of the qnr containing integron among ESBL producing strains may predict future epidemic. Surveillance will be necessary to confirm this low prevalence rate of qnr in France. PMID- 16473480 TI - [Type IV secretion system and their effectors: an update]. AB - Subversion of eukaryotic hosts by bacterial pathogens requires specialized macromolecules secretion systems delivering virulence factors either into the environment or directly into host cells. Transport of molecules across bacterial and eukaryotic membranes is a process requiring multi-component machineries called secretion systems. This review focuses on the Type IV secretion system. This complex is required for genetic exchange (DNA transport) and secretion of effectors (proteins, macromolecules, DNA-proteins complex) into target cells. They transport a wide variety of substrates including large DNA/protein complexes, multi protein toxins, or individual proteins. We describe recent advances on the structure and the function of this secretion system, their effectors and their effects on the functions of eukaryotic cell. PMID- 16473481 TI - Pigeons' memory for sequences of light flashes: reliance on temporal properties and evidence for delay interval/gap confusion. AB - In Experiment 1, pigeons were trained to discriminate between sequences of two and four light flashes (illumination of the feeder). Retention functions obtained with dark delays exhibited a choose-many bias at a 1-s delay and a choose-few bias at delays of 4 and 8s. Retention functions obtained with illuminated delays only displayed a slight choose-few bias. In Experiment 2, additional birds were trained with the same sample sequences. However, the intertrial interval was illuminated by the houselight for Group Light, and it was dark for Group Dark. The acquisition data suggested that multiple temporal features of the light flash sequences controlled choice responding. For both groups, the retention functions were similar to those obtained in Experiment 1. In Experiment 3, baseline training with a 1-s dark delay eliminated the choose-many bias, but a significant choose-few bias at extended dark delays was still observed. Pigeons discriminate light flash sequences by relying on temporal properties of the sequence rather than using an event switch to count flashes. The biased-forgetting effects obtained in these studies appear to be primarily due to confusion between the delay interval and the gap between light flashes. PMID- 16473482 TI - Fetal development of the hand, digits and digit ratio (2D:4D). AB - OBJECTIVE: The purpose of this study was to investigate growth patterns in human hands, digits and digit ratio (2D:4D) during the fetal period. METHODS: The study is carried out on 161 human fetuses (83 males, 78 females) free from external pathology or malformation with ages ranging between 9 and 40 weeks of gestation. Following general external measurements, length and width of the hand, digit lengths separate for each hand was measured, hand index and the ratio of the lengths of the 2nd finger to the 4th finger (2D:4D index) was computed. RESULTS: Means and standard deviations of the parameters with respect to gestational weeks, months and trimesters were calculated. There was a significant correlation between all parameters and gestational age (p<0.001). No significant differences were observed between sexes or sides for any of the parameters (p>0.05). 2D:4D ratio was significantly higher in females compared to males (p<0.05) and mean 2D:4D did not change with gestational age. CONCLUSION: Detailed information of hand and digit parameters related to the fetal period will reveal the extent of biological variations of hand and digit parameters to be used in future studies. We hope that data acquired in this study will facilitate other studies on hand and digit anomalies, pathologies and variations as well as diagnoses and treatments of such conditions conducted in obstetrics, perinatology, forensic medicine and fetal pathology departments. PMID- 16473483 TI - Changes in prostaglandin production and ovarian function in gilts during endometritis induced by Escherichia coli infection. AB - The aim of this study was to determine the prostaglandins (PGs) production and ovarian function in gilts after intrauterine infusions of 10(6) and 10(9) colony forming units (cfu)/ml of Escherichia coli (E. coli). In Experiments 1 and 2, 30 ml of saline or 30 ml of E. coli suspension containing 10(6) or 10(9)cfu/ml, were infused once into each uterine horn in three groups of gilts on day 3 of the estrous cycle, respectively. In Experiment 1, 17 days after treatment it was revealed that inoculation of E. coli 10(9)cfu/ml induced severe acute or subacute endometritis while 10(6)cfu of E. coli evoked moderate acute endometritis or resulted in no inflammatory changes. In the gilts receiving 10(9)cfu/ml of E. coli, the concentration of 13,14-dihydro-15-keto-PGF(2)alpha in blood from the jugular vein was elevated (P<0.05-0.001) compared to concentration in the gilts inoculated with 10(6)cfu on days 8-17 after treatment. Both the E. coli-treated groups had a lower (P<0.05, P<0.01) progesterone plasma level from days 10 to 14 after administration than the control group. On day 17 of the study, infusion of E. coli 10(9)cfu/ml, in comparison to 10(6)cfu, resulted in the greater (P<0.001) content of PGE(2) in the myometrium. The content of both PGs in the endometrium as well as PGF(2alpha) in the myometrium of gilts-treated with 10(9)cfu/ml of E. coli was lower (P<0.001) than in gilts-treated with 10(6)cfu of bacteria. Newly formed corpora lutea were found in the gilts infused with 10(6), but not those infused with 10(9)cfu/ml of E. coli on day 17 after infusion. On day 8 of the study (Experiment 2), the blood from utero-ovarian vein of the gilts-treated with 10(9)cfu/ml of bacteria had a higher (P<0.05) PGF(2alpha) level and lower (P<0.001) PGE(2) level than following infusion of E. coli 10(6)cfu/ml. Also on day 8 of the study, the content of PGE(2) in the endometrium, both the PGs in the myometrium as well as cyclooxygenase-2 in the endometrium and myometrium was greater (P<0.01, P<0.001) after applying 10(9)cfu/ml than 10(6)cfu/ml of E. coli. These results indicate that intrauterine infusions of 10(6) or 10(9)cfu/ml of E. coli lead to the development of inflammatory states of different intensities which is connected with different PGF(2alpha) and PGE(2) production and function of ovaries. PMID- 16473484 TI - The influence of variation of gastric pH on the gelation and release characteristics of in situ gelling pectin formulations. AB - The aim of this study was to examine the influence of variation of gastric pH over the range 1-3 on the gelation of liquid formulations of pectin and on the in vitro and in vivo release of paracetamol and ambroxol from the resultant gels. The formulations were dilute solutions of pectin containing complexed calcium ions that form gels when these ions are released in the acidic environment of the stomach. Gels suitable as vehicles for sustained delivery of these drugs were formed in vitro at pH<3 from pectin solutions of concentrations 1.0-2.0% (w/v). Very weak gels were formed at pH 3.0 resulting in poor sustained release characteristics compared with those at pH 1.2; no significant in vitro gelation was observed at pH 3.5. The bioavailabilities of paracetamol and ambroxol from gels formed in the stomach following oral administration of the liquid formulations were investigated using gastric-acidity controlled rabbits. Visual observations showed in situ gelation of 1.5% (w/v) pectin formulations under conditions of both high (pH 1.0-1.6) and low gastric acidity (pH 3.3-3.6). The bioavailabilities of these drugs were not significantly different when released from gels formed at the two pH limits suggesting that normal variations of gastric acidity in the fasting state will have no effect on the bioavailability of these drugs when delivered using this vehicle. PMID- 16473485 TI - Medicinal plants: an important asset to health care in a region of Central Laos. AB - A field survey of commonly used medicinal plants in the district of Paksan, Bolikhamsai Province in the Lao People's Democratic Republic (Lao P.D.R.) indicates that 55 species of plants, belonging to 49 genera in 31 families of vascular plants, are used in day-to-day medical therapy. Lao names along with uses and preparations for remedies are given. Seven species have medicinal uses that overlap with uses reported in the literature. No medicinal uses have been previously reported for 31 of the species collected based on ethnobotanical field interviews, signifying that their uses may be unique to Laos. Nine of the 31 previously unreported species are mentioned as medicinals multiple times, indicating that further research may be warranted. PMID- 16473486 TI - Ethnobotany of medicinal plants used by Assamese people for various skin ailments and cosmetics. AB - The present paper deals with the medicinal plants used by the people of Assam for curing different skin ailments and for cosmetics. A total of 85 plants belonging to 49 families have been documented for their therapeutic use against skin diseases and as herbal care. The herbal medicines were prepared from various plant parts of single plant, or multiple plants. The majority of the preparation was made using water as the medium. The mode of application was topical, but in many cases it was also administered orally. In several cases the pure herbal preparations was administered along with milk, ghee, honey, coconut oil, curd, etc. Remedies for 18 skin ailments were documented through this study. About 14 plants are known for their use to cure multiple skin diseases. Among these Curcuma longa and Melia azaderach constitute the major plants. The herbal cosmetic products used by the people of Assam ranges from the enhancement of skin colour, hair care, removal of ugly spots, colouring of nails, palms, and teeth. However, many of the plant preparations used for enhancing beauty were also applied for therapeutic use. Herbal remedies were also available for skin burns, prickly heat and pimples. Information on nine plants used for managing dry skin also emerged from this study. PMID- 16473487 TI - Allelic and haplotypic frequencies at 11 Y-STR loci in Buryats from South-East Siberia. AB - We have obtained Y-STR haplotypes in 12 loci (DYS19, DYS385, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438 and DYS439) from 215 Buryat males. We have found that one haplotype (15-11,18-13-28-23-10-11-14-14-10-12) comprises more than 30% of Y chromosomes in this population while another haplotype (14 11,13-14-30-23-10-14-14-14-10-10) comprises additional 14% of chromosomes. The population under study seems to be very homogenous as far as Y chromosome is regarded and the most frequent haplotype seems to be the modal haplotype for Buryats. PMID- 16473488 TI - Optical imaging of lymph nodes. AB - Optical imaging or near infrared fluorescence (NIRF) imaging using enzymatically activatable smart probes is an exiting new imaging modality that can also be used for lymph node visualization and detection. This review intends to briefly summarize general aspects of optical imaging and its capabilities for lymph node imaging. PMID- 16473489 TI - Lymph node imaging: basic principles. AB - Lymph nodes are involved in a wide variety of diseases, particularly in cancer. In the latter, precise nodal staging is essential to guide therapeutic options and to determine prognosis. For long, imaging of the lymphatic system has been limited to lymph vessel,especially via the exclusive use of conventional lymphography, at the expense of invasive procedures and patient's discomfort. Three main technical advances, however, have recently completed the clinical armamentarium for lymph node imaging: first, the refinement of cross sectional imaging, i.e. CT and MRI, combined or not with dedicated contrast agents, has progressively replaced conventional lymphography in oncology situations; second, the development of intra-operative sentinel node mapping has profoundly modified the diagnostic and therapeutic procedures in several cancer situations, mostly melanoma and breast cancer; finally, the increased availability of functional imaging, especially through the use of FDG-PET, has greatly contributed to the accuracy improvement of nodal metastases identification. The aim of this review will thus be to briefly review the anatomy and physiology of the lymphatic systems and to overview the basic principles of up-to-date lymph node imaging. PMID- 16473490 TI - A stability-indicating HPLC assay method for docetaxel. AB - A novel stability-indicating high-performance liquid chromatographic assay method was developed and validated for docetaxel in the presence of degradation products generated from forced decomposition studies. A gradient HPLC method was developed to separate the drug from the degradation products, using a Hichrom RPB HPLC column. Mixture of water and acetonitrile was used as mobile phase. The flow rate was 1.0 ml/min and the detection was done at 230 nm. Using the above method one can carry out the quantitative estimation of impurity namely DCT-1 and docetaxel. The developed gradient LC method was subsequently validated. PMID- 16473491 TI - HPLC determination of calcium pantothenate and two preservatives in topical cream. AB - A RP-HPLC method for simultaneous determination of calcium pantothenate and two preservatives methylparaben and propylparaben present in topical cream was developed. Different analytical columns with various stationary phases were tested. During method development, Supelco Discovery C18 column (125 mmx4.0 mm, 5 microm) and Zorbax SB-CN column (150 mmx4.6 mm, 5 microm) were tested. Both were not convenient for analytical separation because of the co-elution of calcium pantothenate with dead volume, and problems with the peak-shape of all components. Good separation was achieved using Zorbax TSM (250 mmx4.6 mm, 5 microm) and Hypersil ODS column (250 mmx4.6 mm, 5 microm), the latter was finally used for the analysis. The analysis time was 12 min, at flow rate 0.7 ml min-1. Chromatography was performed using binary mobile phase composed of methanol and phosphoric acid, pH 2.5, 65:35 (v/v). UV detection was accomplished at 214 nm. The method was validated according to ICH guideline recommendations. The method is suitable for practical routine analysis of commercially produced topical pharmaceutical preparations. PMID- 16473492 TI - The role of iron in reactive oxygen species generation from diesel exhaust particles. AB - Diesel exhaust particles (DEP) are known to produce reactive oxygen species (ROS), which induce oxidative stress and inflammation in the lung and respiratory tract. DEP are composed of polycyclic aromatic hydrocarbons (PAH) and their derivatives, redox active semi-quinones, and trace amounts of heavy metals. ROS production was measured by thiobarbituric acid-reactive substances of deoxyribose (TBARS) formation from DEP samples obtained from Korea (DEP-KO), and the Standard Reference Material (SRM) 2975 to explore the role of transition metals. Both DEP KO and SRM2975 had similar amounts of transition metals, whereas DEP-KO contained more iron, but less copper and zinc, than SRM2975. The water-soluble fraction from SRM2975, but not that from DEP-KO, had a broad absorption in the visible region, but not from DEP-KO, obscuring an accurate absorption measurement of TBARS. Fluorescence measurements of TBARS generation in a water-soluble extract showed that SRM2975 produces more TBARS, but the addition of hydrogen peroxide (H2O2) generated more TBARS in DEP-KO than in SRM2975, consistent with the higher amounts of iron in DEP-KO. The incubation of DEP with iron chelators inhibited the production of TBARS. Finally, a novel use of the fluorogenic spin trap probe, proxyl fluorescamine, enabled the detection of the ROS production from both DEP KO and SRM2975. Our findings suggested that careful consideration is needed to measure TBARS production in DEP, and that iron in DEP seems to be more important than other transition metals in H2O2-induced ROS generation. PMID- 16473493 TI - A comparative study of using comet assay and gammaH2AX foci formation in the detection of N-methyl-N'-nitro-N-nitrosoguanidine-induced DNA damage. AB - Comet assay is a useful technique in the detection of DNA damages, particularly DNA strand breaks; and it has been utilized to show that a potent carcinogen N methyl-N'-nitro-N-nitrosoguanidine (MNNG), can induce such damages. Recently, gammaH2AX foci formation has been suggested as another sensitive way to detect DNA double strand breaks (DSBs). However, there is no systematic comparison being conducted to evaluate the consistency of these two methods. Using MNNG as a model chemical, the sensitivity of neutral comet assay and gammaH2AX foci formation in detecting MNNG-induced damage was studied. It was found that at concentrations of 0.1 and 1 microg/ml, both methods can detect MNNG-induced damage in human amnion FL cells. However, at 0.1 microg/ml, comet assay revealed more percentage of cells with DNA damage than gammaH2AX fluorescence revealed. On the other hand, while gammaH2AX foci were readily formed at very early times by 10 microg/ml MNNG treatment, neutral comet assay did not detect any significant DNA damage at the same time points. In addition, 10 microg/ml MNNG induced a distinct whole nuclei staining pattern of gammaH2AX, a type of DNA damage which was not detected by neutral comet assay but could be detected by alkaline comet assay. Therefore, gammaH2AX may be used as a sensitive indicator for DNA damage. PMID- 16473494 TI - The subthalamic nucleus: From response selection to execution. AB - The involvement of the subthalamic nucleus in physiological and pathological motor behaviour has now largely been established. Clinical observations in patients suffering from Parkinson disease treated with Deep Brain Stimulation of the STN show that these patients can suffer from postoperative changes in non motor behaviour mainly involving alterations in cognitive functions. The involvement of the STN in cognition has initially been demonstrated by non-human studies investigating the effects of STN lesions and stimulations on cognitive parameters. In the present review, we discuss the findings of these preclinical studies on cognitive parameters and outline the anatomical and functional place of the STN in the basal ganglia cognitive circuit. PMID- 16473496 TI - Behavioral characteristics of children with vocal fold nodules. AB - Vocal fold nodules (VNs) in children are benign, bilateral lesions occurring on the mid-membranous vocal folds. Repetitive phonotraumatic behavior leading to chronic vocal fold injury and repair is frequently cited as the primary etiology; however, specific behavioral characteristics may predispose some children toward intense and potentially phonotraumatic voice use, thereby contributing secondarily to VN formation. The purpose of this case-control study was to determine whether children with VNs possess unique behavioral characteristics that may predispose them to VN development. Parents of 26 children with VNs (20 boys, 6 girls, mean age=7.2 years, SD=2.5 years), and 29 vocally normal, medical controls (22 boys, 7 girls, mean age=6.7 years, SD=2.4), completed the Childhood Behavior Checklist (CBCL/4-18, Achenbach, 1991), a standardized parent-rating scale with strong psychometric properties. No significant between-group differences were detected on any of the behavior problem syndrome scales. Group differences approached significance for the individual items "screams a lot" and "teases a lot" (VN group > Controls). The VN group scored significantly higher than the controls on the "Social Scale," a compilation of positive ratings of the child's social activity, frequency of contacts with friends, behavior with others, and behavior by themselves. Observed outcomes were consistent with previous characterizations of children with VN as "outgoing" or "extroverted" but were not consistent with other claims that this population may be at risk for "aggressive," "attentional," or "impulsive" behavior problems. PMID- 16473495 TI - Effects of prenatal marijuana on visuospatial working memory: an fMRI study in young adults. AB - The long lasting neurophysiological effects of prenatal marijuana exposure on visuospatial working memory were investigated in 18-22 year olds using functional magnetic resonance imaging (fMRI). The participants are members of the Ottawa Prenatal Prospective Study (OPPS), a longitudinal study that provides a unique body of information collected from each participant over 20 years, including prenatal drug history, detailed cognitive/behavioral performance from infancy to young adulthood, and current and past drug usage. This information allowed for the control of potentially confounding drug exposure variables in the statistical analyses. Thirty-one offspring from the OPPS (16 prenatally exposed and 15 nonexposed) performed a visuospatial 2-back task while neural activity was imaged with fMRI. Cognitive performance data were also collected. No significant performance differences were observed when comparing controls versus exposed participants. Multiple regression analyses (including controls with no exposure) revealed that as the amount of prenatal marijuana exposure increased, there was significantly more neural activity in the left inferior and middle frontal gyri, left parahippocampal gyrus, left middle occipital gyrus and left cerebellum. There was also significantly less activity in right inferior and middle frontal gyri. These results suggest that prenatal marijuana exposure alters neural functioning during visuospatial working memory processing in young adulthood. PMID- 16473497 TI - Postoperative parenteral nutrition while proactively minimizing insulin resistance. AB - OBJECTIVE: We compared the metabolic effects of postoperative total parenteral nutrition (TPN) and hypocaloric glucose after treatment with oral carbohydrates preoperatively and epidural anesthesia to proactively minimize postoperative insulin resistance. METHODS: Thirteen patients undergoing colorectal resections were given oral carbohydrates preoperatively and epidural anesthesia and randomized to TPN or hypocaloric glucose during and after surgery. Insulin sensitivity (hyperinsulinemic clamp [0.8 mU x kg(-1) x min(-1)], normoglycemic clamps [4.5 mM]), and glucose kinetics (6,6(2)H2-D-glucose), were studied before and on postoperative day 3. Indirect calorimetry was performed and nitrogen excretion in urine was measured. Values are presented as mean +/- standard deviation. Analysis of variance, planned comparison, and Bonferroni's correction were used for statistical analysis. RESULTS: Three days after surgery insulin stimulated whole-body glucose disposal decreased by 24 +/- 11% versus 28 +/- 23% in patients receiving TPN and hypocaloric glucose, respectively (P < 0.05 for both, not significant between groups). Endogenous glucose production during insulin stimulation was increased only in the glucose group after surgery (P < 0.05 versus before). After surgery, insulin-stimulated glucose oxidation was higher after treatment with TPN, whereas fat oxidation was lower (P < 0.05 for both versus glucose treatment). Fat oxidation increased in the glucose group at basal after surgery (P < 0.05 versus before). Nitrogen balance was less negative after treatment with TPN (P < 0.01). CONCLUSIONS: Treatment with TPN does not seem to improve postoperative peripheral insulin sensitivity in patients with minor insulin resistance after pretreatment with preoperative carbohydrates and perioperative epidural anesthesia. Hypocaloric nutrition results in changes in substrate utilization and nitrogen balance resembling starvation, whereas TPN attenuates these changes. PMID- 16473500 TI - Single staged, Z-plasty supported, wedge excision modification for reconstruction of auricular lesions under local anaesthesia: a technical note. PMID- 16473498 TI - Metastatic tumours to the jaws and oral soft tissues: a retrospective analysis of 41 Korean patients. AB - This article describes a pooled analysis of 41 Korean patients with metastatic oral tumours. The data reviewed are from Korean dental and medical case reports published between 1983 and 2004. The mean age was 55.2 years, and the male-to female ratio was 1.9:1. There were more metastases in the jawbone than in oral soft tissues. The lung was the most common primary site for jawbone metastases, whereas the liver was for those of oral soft tissues. In contrast to reports in Western literature of the breast being the most common primary site, the liver was the most common primary site, followed by the lung and thyroid. These differences may be caused by a relatively high incidence rate of hepatocellular carcinoma in Korea. PMID- 16473502 TI - Interference of antibiotics in the growth curves of oral streptococci. AB - The growth curve interference (GCI) induced by different antibiotics was studied using reference strains of oral streptococci. This parameter may prove to be useful in preventing subacute endocarditis of odontological origin in high-risk patients. The growth curves using different concentrations of antibiotics and with an initial optical density (OD) of 0.05 +/- 0.01 were observed until an OD of 0.9-1.0 was reached. The GCI was defined as the lowest concentration of antibiotic that modified the growth curve with respect to a control without antibiotic. The GCI values were then compared with the minimum inhibitory concentration (MIC) values. In all cases, interference in the growth curves was at least one concentration lower than the MIC in a time ranging from 4-6 h. PMID- 16473501 TI - Benign convulsion with mild gastroenteritis and benign familial infantile seizure. AB - The authors present Japanese siblings of a 6-year-old girl and a 4-year-old boy, who concurrently experienced convulsions with mild gastroenteritis. These siblings, their father and paternal grandfather had afebrile seizures that intermittently occurred without symptoms of gastroenteritis and terminated within a few days at their infancy. An underlying genetic factor might not only cause benign familial infantile seizures but it might also confer the susceptibility to the convulsions with mild gastroenteritis in these siblings. PMID- 16473503 TI - Antibiotic treatment and resistance of unselected uropathogens in the elderly. AB - A population-based study was conducted among women over the age of 70 years with complaints of uncomplicated urinary tract infections (UTIs). The positive predictive value of the nitrite test alone or in combination with the leukocyte esterase test ranged between 83% and 99%. The nitrofurantoin prescription rate decreased whereas fluoroquinolone and amoxicillin/clavulanic acid prescriptions increased with increasing age. The aetiology of infection was age-dependent. Escherichia coli was the most commonly isolated uropathogen, followed by Proteus mirabilis and Klebsiella pneumoniae. For these uropathogens, the lowest susceptibility percentages were found for amoxicillin, trimethoprim and co trimoxazole. As trimethoprim susceptibility reached approximately 75%, it may be advisable not to use this as a first-choice agent in the treatment of uncomplicated UTIs in the elderly. PMID- 16473504 TI - Production and metabolism of indole acetic acid in roots and root nodules of Phaseolus mungo. AB - The mature root nodules of Phaseolus mungo (L.), a leguminous pulse, contain higher amount of indole acetic acid (IAA) than non-nodulated roots. The tryptophan pool present in the mature nodule and young roots might serve as a precursor for the IAA production. Presence of IAA metabolising enzymes - IAA oxidase and peroxidase - indicate the metabolism of IAA in the nodules and roots. In culture, the symbiont, isolated from the nodules, produced a high amount of IAA, when tryptophan was supplied in the medium as a precursor. The symbiont preferred l-isomer over the dl- or d-isomer of tryptophan for IAA production. The important physiological implication of the IAA production in the legume-Rhizobium symbiosis is discussed. PMID- 16473505 TI - Gene regulatory networks and the determination of lymphoid cell fates. AB - Considerable progress has been achieved in the identification and analysis of regulatory molecules that orchestrate B and T cell fate determination. IL-7 and Notch signaling appear to represent analogous pathways that direct B and T cell fates, respectively. For the B lineage, it is possible to assemble the various signaling components and transcription factors into a coherent gene regulatory network. At present, it is not possible to assemble an equivalently precise network for the T lineage as fewer components have been analyzed and their connectivity remains to be more fully explored. However, the ability to manipulate T cell development in a simple culture system using Notch ligand expressing stromal cells is greatly facilitating analysis of this pathway. PMID- 16473506 TI - Point-of-care biosensor systems for cancer diagnostics/prognostics. AB - With the growing number of fatalities resulting from the 100 or so cancer-related diseases, new enabling tools are required to provide extensive molecular profiles of patients to guide the clinician in making viable diagnosis and prognosis. Unfortunately with cancer-related diseases, there is not one molecular marker that can provide sufficient information to assist the clinician in making effective prognoses or even diagnoses. Indeed, large panels of markers must typically be evaluated that cut across several different classes (mutations in certain gene fragments--DNA; over/under-expression of gene activity as monitored by messenger RNAs; the amount of proteins present in serum or circulating tumor cells). The classical biosensor format (dipstick approach for monitoring the presence of a single element) is viewed as a valuable tool in many bioassays, but possesses numerous limitations in cancer due primarily to the single element nature of these sensing platforms. As such, if biosensors are to become valuable tools in the arsenal of the clinician to manage cancer patients, new formats are required. This review seeks to provide an overview of the current thinking on molecular profiling for diagnosis and prognosis of cancers and also, provide insight into the current state-of-the-art in the biosensor field and new strategies that must be considered to bring this important technology into the cancer field. PMID- 16473507 TI - Evaluation of progesterone content in saliva using magnetic particle-based immuno supported liquid membrane assay (m-ISLMA). AB - Progesterone in saliva was monitored using a new method called magnetic particle based immuno supported liquid membrane assay (m-ISLMA) in a sequential injection (SI) setup, allowing automatic sample cleanup, analyte enrichment, and detection in a single analysis unit. Progesterone (Ag) diffuses from a continuous flowing sample - the donor - into a supported organic liquid membrane (SLM), based on analyte partitioning (solubility) between the aqueous donor and the organic phase. The Ag is re-extracted from the SLM into a second stagnant aqueous acceptor, containing antibodies (Ab) immobilized on magnetic beads, held at the bottom of the acceptor by a magnet. Due to the formation of strong Ag-Ab-bead complexes and a large excess of Ab-beads, the Ag is accumulated and selectively enriched in the acceptor. The extracted progesterone was quantified by injecting into the acceptor a horseradish peroxidase (HRP) labeled analyte tracer, the substrate (luminol, H(2)O(2), and p-iodophenol), and finally detection of the generated chemiluminescence by a photomultiplier tube. After optimization of experimental parameters (e.g., sample flow rate, extraction time, type of organic solvent and antibody-bead concentration in the acceptor), a detection limit of 8.50+/-0.17 fgL(-1) and a dynamic range between 35 fgL(-1) and 10 pgL(-1) was reached. The progesterone level of saliva for three subjects (women in different period of ovarian cycle) was investigated, and the corresponding progesterone concentrations detected with m-ISLMA coincided well with the expected values. PMID- 16473508 TI - Expression of steroidogenic factors 1 and 2 in normal human pancreas. AB - Orphan nuclear receptor steroidogenic factor-1 (SF-1) is crucial for development and function of steroidogenic organs. The steroidogenic factor-2 (SF-2) is an essential factor involved in cholesterol transfer and activation of promoters of steroidogenic enzymes CYP11A1, CYP17 and Steroidogenic Acute Regulatory Protein (StAR). We have previously demonstrated steroidogenic activity in pancreatic tissue. The aim of this study was to investigate the presence of SF-1 and SF-2 in human pancreas. Total RNA was extracted from normal male (five) and female (five) samples, obtained from the organs donor program. RT-PCR approach was used to analyze the expression of SF-1 and SF-2. Immunohistochemical analysis was performed for SF-1. The bands of expression were present in both male and female samples, although differential expression was observed. For both factors, the signal detected was more evident in males than in females. A similar pattern was present in the immunohistochemical study. Normal human pancreas expresses SF-1 and SF-2 factors similarly to ovary and adrenals. A distinctive characteristic is the sexually dimorphic expression of these factors. Our data provide evidence suggesting that the pancreas achieves steroidogenic activity supporting the presence of gender- and location-related differences in the expression of these steroidogenic factors. PMID- 16473509 TI - N-thiolated beta-lactams: a new family of anti-Bacillus agents. AB - This report describes the evaluation of N-thiolated beta-lactam antibiotics as potential anti-Bacillus agents. N-Thiolated beta-lactams are a new family of antibacterials that previously have been found to selectively inhibit the growth of Staphylococcus bacteria over many other genera of microbes. From the data presented herein, these lactams similarly inhibit a variety of Bacillus species, including Bacillus anthracis. The preliminary structure-activity studies suggest that there is a need to balance the lipophilic character of the C3/C4 groups in order to obtain optimal anti-Bacillus activity. Elongation or extensive branching of the organothio substituent diminishes antibacterial effects, with the sec butylthio derivative providing the strongest activity. PMID- 16473510 TI - Amyloids, prions and the inherent infectious nature of misfolded protein aggregates. AB - Misfolded aggregates present in amyloid fibrils are associated with various diseases known as "protein misfolding" disorders. Among them, prion diseases are unique in that the pathology can be transmitted by an infectious process involving an unprecedented agent known as a "prion". Prions are infectious proteins that can transmit biological information by propagating protein misfolding and aggregation. The molecular mechanism of prion conversion has a striking resemblance to the process of amyloid formation, suggesting that misfolded aggregates have an inherent ability to be transmissible. Intriguing recent data suggest that other protein misfolding disorders might also be transmitted by a prion-like infectious process. PMID- 16473511 TI - Bilateral stress fracture of the patellae: a case report. AB - A case of bilateral simultaneous stress fracture of the patella is presented in a 64 year old Marathon runner sustained when he slipped off a curb whilst out walking. This was confirmed on Xray and treated by tension band wiring using the AO technique. He had an excellent result returning to normal function in both knees. This case is unusual since the fractures were not due to osteoporosis, as would be expected in this age group. PMID- 16473512 TI - Sustained astrocytic clusterin expression improves remodeling after brain ischemia. AB - Clusterin is a glycoprotein highly expressed in response to tissue injury. Using clusterin-deficient (Clu-/-) mice, we investigated the role of clusterin after permanent middle cerebral artery occlusion (MCAO). In wild-type (WT) mice, clusterin mRNA displayed a sustained increase in the peri-infarct area from 14 to 30 days post-MCAO. Clusterin transcript was still present up to 90 days post ischemia in astrocytes surrounding the core infarct. Western blot analysis also revealed an increase of clusterin in the ischemic hemisphere of WT mice, which culminates up to 30 days post-MCAO. Concomitantly, a worse structural restoration and higher number of GFAP-reactive astrocytes in the vicinity of the infarct scar were observed in Clu-/- as compared to WT mice. These findings go beyond previous data supporting a neuroprotective role of clusterin in early ischemic events in that they demonstrate that this glycoprotein plays a central role in the remodeling of ischemic damage. PMID- 16473513 TI - Minimally invasive direct coronary artery bypass versus stenting. PMID- 16473514 TI - Coronary artery size and disease in UK South Asian and Caucasian men. AB - OBJECTIVE: South Asian patients in the UK have a higher mortality rate after coronary artery bypass grafting (CABG) than Caucasian patients. As coronary artery size has been shown to correlate to outcome from bypass grafting, it has been suggested that smaller coronary arteries in South Asians as compared to Caucasians could contribute to a poorer outcome in the Asian population. We aimed to measure coronary artery size and disease in matched South Asian and Caucasian men undergoing first time coronary artery bypass grafting. METHODS: Coronary arteriograms from 53 matched first generation South Asian and Caucasian men were examined. The patients had no history of myocardial infarction, coronary revascularisation, familial dyslipidaemia, diabetes or renal disease. They were individually matched for age, height, weight, body mass index and body surface area. Thereafter, coronary artery diameters and significant (> or =50%) diameter stenoses were measured in a blinded fashion using quantitative coronary angiography (QCA). RESULTS: In South Asian men, diameters of the left main stem (LMS) and the proximal left anterior descending, the circumflex and the right coronary arteries were 4.6+/-0.9 mm, 3.5+/-0.8 mm, 3.4+/-0.8 mm and 3.5+/-0.8 mm, respectively. The corresponding arterial diameters among Caucasian men (4.5+/-0.9 mm, 3.5+/-0.7 mm, 3.5+/-0.8 mm and 3.8+/-0.8 mm) did not differ from those in South Asians. There was no difference in the number of significant coronary artery stenoses between the two groups and no difference in bypass and cross clamp times or in adverse outcome (one from each group died after coronary artery bypass grafting). CONCLUSION: Proximal coronary artery size and number of significant coronary stenoses did not differ between matched pairs of South Asian and Caucasian men using strict inclusion/exclusion criteria. PMID- 16473515 TI - Aortobrachial bypass for acute vascular insufficiency due to tumor embolization. PMID- 16473516 TI - The vacuum bell for treatment of pectus excavatum: an alternative to surgical correction? AB - OBJECTIVE: Pectus excavatum (PE) is the most common chest wall malformation and one of the most frequent major congenital anomalies. The surgical repair of PE in childhood is a well-established procedure. Previously used operative techniques to correct PE were largely based on the Ravitch technique. Today, the minimally invasive repair (MIRPE) by Nuss is well established. Conservative treatment with the vacuum bell to elevate the funnel in patients with PE represents a potential alternative. METHODS: A suction cup is used to create a vacuum at the anterior chest wall. A patient-activated hand pump is used to reduce the pressure up to 15% below atmospheric pressure. Three different sizes of vacuum bell exist which are selected according to the individual patients age. When creating the vacuum, the lift of the sternum is obvious and remains for a different time period. The device should be used for a minimum of 30 min (2 per day), and may be used up to a maximum of several hours daily. Presently, a 12-15-month course of treatment is recommended. In addition, the device was used intraoperatively during the MIRPE procedure to enlarge the retrosternal space to ensure safer passage of the introducer in a few patients. RESULTS: Thirty-four patients (31 males, 3 females), aged 6-52 years (median 17.8 years) used the vacuum bell for 1 to maximum 18 months (median 10.4 months). Follow-up included photography and clinical examination every 3 months. Computed tomographic scans showed that the device lifted the sternum and ribs immediately. In addition, this was confirmed thoracoscopically during the MIRPE procedure. After 3 months, an elevation of more than 1.5 cm was documented in 27 patients (79%). After 12 months, the sternum was lifted to a normal level in five patients (14.7%). Relevant side effects were not noted. CONCLUSIONS: The vacuum bell has proved to be an alternative therapeutic option in selected patients with PE. The initial results proved to be dramatic, but long-term results are so far lacking, and further evaluation and follow-up studies are necessary. In addition, the method may assist the surgeon during the MIRPE procedure. PMID- 16473517 TI - Factors affecting survival after bronchoplasty and broncho-angioplasty for lung cancer: single institutional review of 147 patients. AB - OBJECTIVE: We compared the results of bronchoplasty and bronchoplasty with pulmonary angioplasty (broncho-angioplasty). Multivariate analysis was done to determine potential prognostic factors for both procedures based on our 20-year single-center experience. METHODS: Between January 1985 and December 2004, 163 bronchoplastic procedures were done in 1405 patients who underwent lung resections for lung cancer at Nagasaki University Hospital. After excluding 16 carino-plasty patients, 147 patients (118 bronchoplasty and 29 broncho angioplasty) were included. RESULTS: In the bronchoplasty group, the postoperative morbidity was 22.9% (27/118) and the 90-day postoperative mortality was 5.9% (7/118), while in the broncho-angioplasty group the postoperative morbidity was 27.6% (8/29) and the postoperative mortality was 17.2% (5/29). The 5-year survival for all patients was 56.0%. Among patients with stage I or II, the 5-year survival was 76.2% in the bronchoplasty group and 51.9% in the broncho angioplasty group (p=0.1791). On the other hand, among patients with stage III or IV, 5-year survival was 43.5% in the bronchoplasty group and 7.9% in the broncho angioplasty group (p=0.0192). Multivariate analysis indicated that the type of operation, postoperative complications, histologic type, and pN status were significant factors affecting survival. CONCLUSIONS: Both bronchoplasty and broncho-angioplasty are useful for the treatment of patients with lung cancer and should be performed in stage I or II. However, careful patient selection is mandatory in patients with advanced tumor stages and in those with nonsquamous cell carcinoma, especially if broncho-angioplasty is being considered. PMID- 16473518 TI - Chest wall mass with double pathology. AB - Intrathoracic neurogenic tumors arising from chest wall are generally rare tumors. The benign soft tissue tumors may produce compression effect on the chest wall but are generally free. The presence of unusual adherence raises the suspicion of malignancy. Our case report describes the clinical features of a young male who underwent excision of a left posterosuperior chest wall mass with a portion of the fourth rib. Histopathological examination unexpectedly revealed the existence of two different pathologies. The mass was found to be benign schwannoma and the rib showed features of tuberculous osteomyelitis. Inflammatory response and fibrous reaction mimicked the features of malignancy. PMID- 16473519 TI - Validation of the EuroSCORE model in Australia. AB - OBJECTIVE: There is an important role for accurate risk prediction models in current cardiac surgical practice. Such models enable benchmarking and allow surgeons and institutions to compare outcomes in a meaningful way. They can also be useful in the areas of surgical decision-making, preoperative informed consent, quality assurance and healthcare management. The aim of this study was to assess the performance of the European System for Cardiac Operative Risk Evaluation (EuroSCORE) model on the Australasian Society of Cardiac and Thoracic Surgeons (ASCTS) patient database. METHODS: The additive and logistic EuroSCORE models were applied to all patients undergoing cardiac surgery at six institutions in the state of Victoria between 1st July 2001 and 4th July 2005 within the ASCTS database who have complete data. The entire cohort and a subgroup of patients undergoing coronary artery bypass grafting (CABG) only were analysed. Observed and predicted mortalities were compared. Model discrimination was tested by determining the area under the receiver operating characteristic (ROC) curve. Model calibration was tested by the Hosmer-Lemeshow chi-square test. RESULTS: Eight thousand three hundred and thirty-one patients with complete data were analysed. There were significant differences in the prevalence of risk factors between the ASCTS and European cardiac surgical populations. Observed mortality was 3.20% overall and 2.00% for the CABG only group. The EuroSCORE models over estimated mortality (entire cohort: additive predicted 5.31%, logistic predicted 8.76%; CABG only: additive predicted 4.25%, logistic predicted 6.19%). Discriminative power of both models was very good. Area under ROC curve was 0.83 overall and 0.82 for the CABG only group. Calibration of both models was poor as mortality was over predicted at nearly all risk deciles. Hosmer-Lemeshow chi-square test returned P-values less than 0.05. CONCLUSIONS: The additive and logistic EuroSCORE does not accurately predict outcomes in this group of cardiac surgery patients from six Australian institutions. Hence, the use of the EuroSCORE models for risk prediction may not be appropriate in Australia. A model, which accurately predicts outcomes in Australian cardiac surgical patients, is required. PMID- 16473520 TI - Capecitabine use in geriatric oncology: an analysis of current safety, efficacy, and quality of life data. AB - Breast and colon cancer occur primarily in people over the age of 65 years, yet standards of care are mostly derived from clinical trials conducted with predominantly younger patients. Clinicians caring for older patients largely rely on clinical judgment and anecdotal experience and most would agree that additional research is needed to optimize care for this growing subset of patients. Capecitabine, a fluoropyrimidine formulated for oral administration has theoretical appeal in this population. In this analysis of published reports and recent presentations published in abstract form, the data on capecitabine efficacy and toxicity are reviewed in the context of patient age. For both breast and colon cancer, capecitabine alone or in combination with other cytotoxics is safe and effective. PMID- 16473521 TI - Effects of gabexate mesilate on serum inflammatory cytokines in rats with acute necrotizing pancreatitis. AB - Gabexate mesilate is a synthetic protease inhibitor. The effectiveness of gabexate mesilate in patients with acute pancreatitis is controversial. Proinflammatory cytokines are associated with systemic inflammatory response syndrome (SIRS) in acute pancreatitis. A compensatory anti-inflammatory response occurs in parallel with SIRS. We investigated the effects of gabexate mesilate on acute necrotizing pancreatitis in rats, emphasizing the changes in serum levels of proinflammatory and anti-inflammatory cytokines. Acute necrotizing pancreatitis was induced by retrograde infusion of sodium taurodeoxycholate into the pancreatobiliary duct in rats. The rats were divided into three groups. Group I was given gabexate mesilate 2 mg/kg/h i.v. continuously 1 h before the induction of acute pancreatitis. Group II was given gabexate mesilate the same dose immediately after the induction of acute pancreatitis. Group III was given normal saline as the controls. Serum levels of amylase, lipase, tumor necrosis factor alpha, interleukin-6, and interleukin-10, pancreatic histopathology and hemodynamics were examined at 5h after the induction of acute pancreatitis. Gabexate mesilate significantly reduced serum levels of amylase, lipase, tumor necrosis factor alpha and interleukin-6 at 5 h. Serum levels of interleukin-10 significantly increased in Group I, as compared with Groups II and III. The severity of pancreatic histopathology, the reduction of mean arterial pressure, the volume of ascites and pancreatic wet weight/body weight ratios were also significantly improved by the administration of gabexate mesilate. The beneficial effects of gabexate mesilate on acute pancreatitis may be, in part, due to the modulation of inflammatory cytokine responses. PMID- 16473522 TI - Decreased frequency of the TNF2 allele of TNF-alpha -308 promoter polymorphism is associated with lacunar infarction. AB - BACKGROUND AND PURPOSE: Enhanced release of proinflammatory cytokines may contribute to the pathogenesis of stroke. It was examined whether G to A promoter polymorphism in the tumor necrosis factor-alpha gene at position -308 affects the risk of stroke. METHODS: We genotyped 336 patients with ischemic stroke and 333 healthy controls for this polymorphism. Patients were divided into different groups based on the Oxfordshire Community Stroke Project (OCSP) or a modified TOAST classification. Distribution of the alleles at -308 G>A promoter polymorphism was determined by PCR-RLFP method. RESULTS: Patients with ischemic stroke had a significantly (p<0.001) decreased (0.115) frequency of the -308 A (TNF2) allele compared to the healthy controls (0.196). When patients were categorized according to the OCSP classification, it turned out that significant (p=0.002) decrease in TNF2 allele frequency (0.065) was restricted to the patients with lacunar infarct (LACI) whereas the frequency of the TNF2 alleles in patients with the other three subtypes (TACI, PACI, and POCI) did not significantly differ from that in healthy controls. Similar results were obtained when the patients were divided according to the modified TOAST classification: the frequencies of the TNF2 allele were 0.068 and 0.140 (p=0.010) in the patients with small-vessel and non-small vessel (large vessel infarction or ischemic stroke of other origin) infarction, respectively. The age-adjusted odds ratio of the patients carrying the TNF2 allele to develop lacunar infarct was 0.33 (0.16 0.68) (p=0.002) compared to the non-carriers. This difference was also restricted to the male patients. CONCLUSIONS: Our results suggest that male carriers of TNF2 allele are less susceptible for the development of lacunar subtype of ischemic stroke than the non-carriers. PMID- 16473523 TI - Instruction of Treg commitment in peripheral T cells is suited to reverse autoimmunity. AB - In order to exploit regulatory T cells in a clinical setting it is desirable to be able to generate such cells by a variety of antigens that elicit unwanted immune responses. This goal has been achieved by targeting antigen to dendritic cells under subimmunogenic conditions which results in the conversion of naive Foxp3 negative T cells into Foxp3-expressing regulatory T cells that are indistinguishable from what has been referred to as natural Treg. Such cells have the ability to interfere with immunity at early as well as late stages of the immune response during which effector cells have already been formed. This suggests that Treg cannot only be exploited to prevent immune responses but also to interfere with already established immunity. PMID- 16473524 TI - Measurement of cytokine release at the single cell level using the ELISPOT assay. AB - The enzyme-linked immunospot (ELISPOT) assay took its design concept from traditional ELISA techniques and evolved over the years from a method for detecting antibodies secreted from B cells to a method for detecting cytokines or other soluble mediators secreted from a variety of different cell types. The ELISPOT assay allows the quantitative measurement of frequency of cytokine secreting cells at the single cell level directly ex vivo without elaborate in vitro expansion or manipulation of cell populations. The function of cells can be inferred from the pattern of cytokines secreted by cells in response to diverse antigenic stimuli and thus the ELISPOT assay has become a powerful method for monitoring immune responses in health and disease. The ELISPOT assay like the ELISA assay is relatively easy to perform and it has the promise of robustness, reliability, and reproducibility of performance for use as a diagnostic tool. The history, applications, validation process, and future challenges of the ELISPOT assay are discussed in this chapter. PMID- 16473525 TI - High-level expression and purification of human thymidine kinase 1: quaternary structure, stability, and kinetics. AB - Human cytosolic thymidine kinase (hTK1) is the key enzyme of the pyrimidine salvage pathway and phosphorylates thymidine to thymidine monophosphate, a precursor building block of the DNA. Wild-type hTK1 (hTK1W) as well as a truncated form of the enzyme (hTK1M) carrying deletions at the N- and C-terminal regions were cloned as His(6)-tagged fusion proteins. Expression, isolation, and purification protocols have been established, leading to high yields of soluble and active wild type (approximately 35 mg) and truncated hTK1 (approximately 23 mg) per liter of culture. The protein was purified to near homogeneity. The chaperone DnaK was identified to be the major contaminant that could be removed by applying an additional ATP-MgCl(2) incubation and washing step. hTK1W was a permanent tetramer in solution, whereas the truncated construct hTK1M appears to be a dimer in absence and presence of substrates. Both hTK1W and hTK1M exhibit pronounced thermal stability with transition temperatures (T(m)) of 71.7 and 73.4 degrees C, respectively, when measured without adding substrates. The presence of substrates stabilized both hTK1W (DeltaT(m) ranging from 5.6 to 12.5 degrees C) and hTK1M (DeltaT(m) ranging from 0.8 to 5.3 degrees C). Both enzymes show high activity over a broad range of pH, temperature, and ionic strength. Kinetic studies determined a K(M) of 0.51 microM and a k(cat) of 0.28 s(-1) for wild-type hTK1. The truncated hTK1M has a K(M) of 0.87 microM and k(cat) of 1.65 s(-1), thus exhibiting increased catalytic efficiency. The availability of recombinant human TK1 will facilitate further biochemical and crystallographic studies. PMID- 16473526 TI - Cultural variability in the effects of question design features on respondent comprehension of health surveys. AB - PURPOSE: The aim is to identify the characteristics of common health survey questions that may be associated with cross-cultural variability in question comprehension. METHODS: Interviews with respondents representing four cultural subgroups in the United States were analyzed through behavior coding to identify survey question characteristics that may be associated with cross-cultural variability in comprehension. Using survey responses as the unit of analysis, nested within survey respondents and survey questions, hierarchical linear modeling was used to examine the effects of four questionnaire design features on cultural variations in comprehension difficulties. RESULTS: Respondent culture was found to moderate the effects of response format, question length, and reading levels. CONCLUSIONS: Several question design strategies that reduce overall comprehension difficulty appear to also increase cross-cultural disparities. PMID- 16473527 TI - Correlative microXRF and optical immunofluorescence microscopy of adherent cells labeled with ultrasmall gold particles. AB - Synchrotron-based X-ray fluorescence microscopy (microXRF) is a powerful tool to study the two-dimensional distribution of a wide range of biologically relevant elements in tissues and cells. By growing mouse fibroblast cells directly on formvar-carbon coated electron microscopy grids, microXRF elemental maps with well-defined subcellular resolution were obtained. In order to colocalize the elemental distribution with the location of specific cellular structures and organelles, we explored the application of a commercially available secondary antibody conjugated to FluoroNanogold, a dual-label that combines a regular organic fluorophore with a 1.4 nm Au-cluster as xenobiotic label for microXRF imaging. Adherent mouse fibroblast cells were grown on silicon nitride windows serving as biocompatible XRF support substrate, and labeled with FluoroNanogold in combination with primary antibodies specific for mitochondria or the Golgi apparatus, respectively. Raster scanning of the in-air dried cells with an incident X-ray energy of 11.95 keV, sufficient to ensure excitation of the Au Lalpha line, provided two-dimensional maps with submicron resolution for Au as well as for most biologically relevant elements. MicroXRF proved to be sufficiently sensitive to image the location and structural details of the Au labeled organelles, which correlated well with the subcellular distribution visualized by means of optical fluorescence microscopy. PMID- 16473528 TI - Transcranial magnetic stimulation and stroke: a computer-based human model study. AB - This paper explores how transcranial magnetic stimulation (TMS) induced currents in the brain are perturbed by electrical and anatomical changes following a stroke in its chronic stage. Multiple MRI derived finite element head models were constructed and evaluated to address the effects that strokes can have on the induced stimulating TMS currents by comparing stroke models of various sizes and geometries to a healthy head model under a number of stimulation conditions. The TMS induced currents were significantly altered for stimulation proximal to the lesion site in all of the models analyzed. The current density distributions were modified in magnitude, location, and orientation such that the population of neural elements that are stimulated will be correspondingly altered. The current perturbations were minimized for conditions tested where the coil was far removed from the lesion site, including models of stimulation contralateral to the lesioned hemisphere. The present limitations of TMS to the peri-lesional cortex are explored, ultimately concluding that conventional clinical standards for stimulation are unreliable and potentially dangerous predictors of the site and degree of stimulation when TMS is applied proximal to infarction site. PMID- 16473529 TI - History of the Crested Lark in the Mediterranean region as revealed by mtDNA sequences and morphology. AB - The Crested Lark has a very complex taxonomy, partly as a result of a strong variation in plumage ground color seemingly linked with environmental factors. However, large variations in body size and bill shape further complicate the situation in the Maghreb. In this paper, we first present a set of hypotheses to explain patterns of morphological variation around the Mediterranean Sea. A phylogeographical analysis covering all major biogeographical areas in the species' range is then performed to test these scenarios. Three mtDNA groups with distinct geographical distribution were identified. The randonii clade (= G. (c.) randonii) is endemic from central Maghreb and is phylogenetically basal relative to cristata and senegallensis. These two latter groups are much more widespread. The cristata clade is found in NW Morocco, throughout Europe and W Asia and in NE Africa, while senegallensis regroups the populations sampled in the Western Sub Saharan Africa and in NE Maghreb (E Algeria, Tunisia). A combination of genetic and paleoenvironmental evidences supports a scenario of allopatric differentiation of these two lineages outside the Maghreb, with subsequent range expansion leading to their secondary presence in the Maghreb. However, the alternative hypothesis of differentiation in two, or even three separate Maghreb refuges cannot be completely dismissed with the present data. Interestingly, the Sahara desert and the Gibraltar Strait did not act as permanent barriers to dispersal in this species. In addition, the populations in the Maghreb are consistently longer-billed than their closest relatives, suggesting a role for natural selection or phenotypic plasticity. PMID- 16473530 TI - Analysis of the complete mitochondrial DNA from Anopheles funestus: an improved dipteran mitochondrial genome annotation and a temporal dimension of mosquito evolution. AB - Virtually no information regarding timing of deep lineage divergences within mosquito family (Culicidae) exists, which poses an important problem in the postgenomic era. To address this issue, the complete 15,354 bp mitochondrial genome of Anopheles funestus was assembled from both mtDNA and cDNA sequences generated from transcripts of the mtDNA-encoded protein and rRNA genes. Analysis of the transcript information allowed an improved genome annotation, revealing that the translation initiation codon for the cox1 gene is TCG, rather than atypical, longer codons proposed in several other insects. The 5'ends of nad1 and nad5 transcripts begin with TTG and GTG triplets, respectively, which apparently serve as the translation initiators for those genes. We used all the A. funestus mtDNA gene sequences and three other publicly available mosquito mtDNA genomes for the estimation of divergence time points within Culicidae. The maximum likelihood date estimates for the splits between Anopheles and Aedes (approximately 145-200 Mya), between Anopheles subgenera Cellia and Anopheles (approximately 90-106 Mya), and between lineages within subgenus Anopheles (approximately 70-85 Mya) inferred from protein-coding genes are roughly twice as high as the dates based on RNA gene sequences. Although existing evidence does not unequivocally favor one of the alternatives, fossil-based predictions of the age of the family Culicidae are in better agreement with dates inferred from protein-coding genes. PMID- 16473531 TI - Enhancement of long-term memory retention by Colostrinin in one-day-old chicks trained on a weak passive avoidance learning paradigm. AB - Colostrinin (CLN) is a biologically active proline-rich polypeptide which has therapeutic potential for the alleviation of memory deficits in age-related dementias in a number of human conditions, particularly Alzheimer's disease. To examine the efficacy of CLN in other species, day-old domestic chicks were used as a model system to study its effects on retention of memory for a single one trial learning paradigm--avoidance of a bitter-tasting substance (methylanthranilate, MeA). Birds were presented with a bead coated with either a dilute (10%) solution of MeA or a bead coated with 100% MeA. Those trained on 100% MeA avoided pecking at a similar but dry bead 24 h later, thereby demonstrating long-term memory whereas chicks trained on the 10% solution pecked the bead at 24 h, indicating lack of long term memory for the task. However, when CLN was injected (i.c.) into a region known to be important in memory formation, the mesopallium intermediomediale (IMM), prior to training with 10% MeA, chicks exhibited strong memory retention at 24 h, similar to those trained on 100% MeA. Control chicks trained on 10% MeA but injected i.c. with a 10% saline solution did not show improvement in memory retention. Intraperitoneal (i.p.) injections of CLN were as effective as the i.c. route. These data extend the known efficacy of CLN from mammals demonstrating its widespread efficacy as a cognitive enhancer. PMID- 16473533 TI - Novel targets for antithrombotic drug discovery. AB - Platelet aggregation is a dynamic entity, capable of directing its own growth and stability via the activation of signaling cascades that lead to the expression and secretion of various secondary agonists. Recent data using proteomics and genomics strategies have established that signaling pathways during platelet aggregation are triggered by two homophilic adhesion molecules, CD84 and CD150 (SLAM), and by a novel EGF-containing receptor, PEAR1, which are tyrosine phosphorylated in a platelet-aggregation-dependent fashion (N. Nanda, P. Andre, M. Bao et al., Platelet aggregation induces platelet aggregate stability via SLAM family receptor signaling, Blood 106 (2005) 3028-3034, N. Nanda, M. Bao, H. Lin et al., Platelet Endothelial Aggregation Receptor 1 (PEAR1), a novel epidermal growth factor repeat-containing transmembrane receptor, participates in platelet contact-induced activation, J. Biol. Chem. 280 (2005) 24680-24689). Analysis of SLAM-deficient mice revealed an overall defect in platelet aggregation in vitro and a delayed arterial thrombotic process in vivo. The data indicate that these aggregation co-receptors may function in a "platelet synapse" and may be novel targets for antithrombotic drug discovery. PMID- 16473534 TI - Contact-dependent signaling events that promote thrombus formation. AB - There is increasing evidence that formation of a stable hemostatic plug requires adhesive and signaling events that continue beyond the onset of platelet aggregation. These events are facilitated and, in some cases, made possible, by the persistent close contacts between platelets that can only occur when platelets begin to aggregate. Participants include integrins and other cell adhesion molecules, secreted agonists, receptor tyrosine kinases, and protein fragments that are shed from the surface of activated platelets. Collectively, these molecules promote the continued growth and stability of the hemostatic plug. PMID- 16473535 TI - Initiation of coagulation by tissue factor carriers in blood. AB - Fibrin can be generated in blood vessels in the absence of substantial damage to the vessel wall. According to novel observations, fibrin formation within the vessel lumen could be initiated by intravascular (blood borne) tissue factor (TF), a central starter protein of blood coagulation. We have recently detected TF in platelets, principally allowing coagulation to be initiated within the developing thrombus. While TF is stored in intraplatelet compartments under resting conditions, it is rapidly translocated to the cell surface in response to platelet activation, accumulating on filopodia. Platelet TF might be acquired from extracellular sources via transfer of TF positive microparticles and/or, potentially, be generated through translational mechanisms. Exocytotic microparticles, regular blood components, share the ability of the activated platelets to coexpose TF and phosphatidylserine, allowing the assembly of the entire coagulation system on a single membrane surface. Nonetheless, the procoagulant activity of the microparticles, when present alone, is limited. However, the vesicular TF might be uncovered by platelet-microparticle interactions. Thereby, the recruitment of platelets and microparticles to the site of vascular injury could synergistically trigger fibrin generation. In summary, by utilizing differential pathways, activated platelets are mandatory for the TF-mediated coagulation start in blood. PMID- 16473536 TI - Purification, cDNA cloning and homology modeling of endo-1,3-beta-D-glucanase from scallop Mizuhopecten yessoensis. AB - The retaining endo-1,3-beta-D-glucanase (LV) with molecular mass of 36 kDa was purified to homogeneity from the crystalline styles of scallop Mizuhopecten yessoensis. The purified enzyme catalyzed hydrolysis of laminaran as endo-enzyme forming glucose, laminaribiose and higher oligosaccharides as products (Km approximately 600 microg/mL). The 1,3-beta-D-glucanase effectively catalyzed transglycosylation reaction that is typical of endo-enzymes too. Optima of pH and temperature were at 4.5 and 45 degrees C, respectively. cDNA encoding the endo 1,3-beta-D-glucanase was cloned by PCR-based methods. It contained an open reading frame that encoded 339-amino acids protein. The predicted endo-1,3-beta-D glucanase amino acid sequence included a characteristic domain of the glycosyl hydrolases family 16 and revealed closest homology with 1,3-beta-D-glucanases from bivalve Pseudocardium sachalinensis, sea urchin Strongylocentrotus purpuratus and invertebrates lipopolysaccharide and beta-1,3-glucan-binding proteins. The fold of the LV was more closely related to kappa-carrageenase, agarase and 1,3;1,4-beta-D-glucanase from glycosyl hydrolases family 16. Homology model of the endo-1,3-beta-D-glucanase from M. yessoensis was obtained with MOE on the base of the crystal structure of kappa-carrageenase from P. carrageonovora as template. Putative three-dimensional structures of the LV complexes with substrate laminarihexaose or glucanase inhibitor halistanol sulfate showed that the binding sites of the halistanol sulfate and laminarihexaose are located in the enzyme catalytic site and overlapped. PMID- 16473537 TI - A genetic redox sensor for mammalian cells. AB - Nutrient and oxygen availability are key metabolic parameters for biopharmaceutical manufacturing. In order to enable mammalian cells to manifest their intracellular nutrient and oxygen levels we engineered a genetic sensor circuitry which converts signals impinging on the cellular redox balance into a robust reporter gene expression readout. Capitalizing on the Streptomyces coelicolor redox control system, consisting of REX modulating ROP-containing promoters in an NADH-dependent manner, we designed a mammalian dual sensor transcription control system by fusing REX to the generic VP16 transactivation domain of Herpes simplex, which reconstitutes an artificial transactivator (REDOX) able to bind and activate chimeric promoters assembled by placing a ROP operator module 5' of a minimal eukaryotic promoter (P(ROP)). When nutrient levels were low and resulted in depleted NADH pools REDOX-dependent P(ROP)-driven expression of secreted (human-secreted alkaline phosphatase; SEAP) or intracellular (Renilla reniformis luciferase; rLUC) reporter genes was high as a consequence of increased REDOX-P(ROP) affinity. Conversely, at hypoxic conditions leading to high intracellular NADH levels, strongly reduced REDOX-P(ROP) interaction mediated low-level transgene expression in Chinese hamster ovary (CHO K1) cells. Other molecules (for example, 2,4-dinitrophenol, cyanide or hydrogen peroxide) which are known to imbalance the intracellular NADH/NAD+ poise could also be detected using the REDOX-P(ROP) sensor circuitry. REDOX's sensor capacity (nutrient and oxygen levels) operated seamlessly in transgenic CHO-K1 cell derivatives adapted for growth in serum-free suspension cultures and enabled precise monitoring of the population's metabolic state. As the first genetic metabolic sensor designed for mammalian cells, REDOX may foster advances in process development and biopharmaceutical manufacturing. PMID- 16473538 TI - Primary and opportunistic pathogens associated with meningitis in adults in Bangui, Central African Republic, in relation to human immunodeficiency virus serostatus. AB - OBJECTIVE: To determine the causative organisms and characteristics of patients presenting with meningitis in Bangui in order to provide guidance to physicians for case management. METHODS: Adults with proven or suspected meningitis were enrolled in this prospective study. LABORATORY TESTS: Full blood count, blood chemistry, and HIV tests were performed. Cerebrospinal fluid (CSF) was submitted for routine microbiology, chemistry (glucose, protein), and hematology testing. When classical microbiology analyses were negative, a broad-range bacterial polymerase chain reaction (BRBPCR) was used. RESULTS AND CONCLUSIONS: Of the 276 patients enrolled, 215 (77.9%) were HIV positive. In HIV-positive patients cryptococcal meningitis (CM) was the most common cause of meningitis (39.1%) followed by pyogenic meningitis (PM) (30.7%), mononuclear meningitis (MM) (28.8%), and tuberculous meningitis (TM) (1.4%). In HIV-negative patients, PM was the most common cause (60.7%) followed by MM (37.7%) and CM (1.6%, one case). In hospital mortality was higher in HIV-positive patients (73/128 = 57%) compared to those HIV negative (3/18 = 16.7%) (p = 0.001). Streptococcus pneumoniae (n = 26) was the most common bacterial diagnosis, mainly in HIV-positive patients (n = 22, 10.2%). Meningococcal meningitis (14 Neisseria meningitidis of group A and one W135) was diagnosed in nine (4.2%) HIV-positive and six (9.8%) HIV-negative patients. Gram-negative rods were isolated from five HIV-positive and two HIV negative patients, respectively. The bacteria and fungi involved in meningitis did not display high levels of in vitro resistance. Conventional microbiology techniques failed to detect the causative agent in 55 (53.4%) PM cases. Broad range bacterial PCR detected DNA from S. pneumoniae in three samples, N. meningitidis in two, Escherichia coli in one, Listeria monocytogenes in two and Staphylococcus aureus in one sample. In the CSF of five (three HIV negative and two HIV positive), PCR products were not identified with the oligonucleotide probes specific for the usual species of bacteria found in CSF, or genera commonly considered potential contaminants of clinical samples. Among the MM cases, 77 (90.5%) probable viral meningitis (54 HIV positive and 23 HIV negative) and eight TM (HIV positive) were suspected. PMID- 16473539 TI - Helicobacter pylori regulates the expression of inhibitors of DNA binding (Id) proteins by gastric epithelial cells. AB - Id transcription factors control proliferation, differentiation and apoptosis by inhibiting the DNA binding of basic helix-loop-helix transcription factors. Increased expression of Id proteins promotes proliferation, inhibits differentiation, and is associated with intestinal tumorigenesis. We aimed to determine how Helicobacter pylori may alter the expression of Id proteins by gastric epithelial cells: it was hypothesised that H. pylori, a known carcinogen, would result in increased expression of one or more Id family members. In vitro and in vivo models of infection were employed, including treatment of AGS gastric epithelial cells with wild-type H. pylori strains, 60190 and SS1, and Mongolian gerbils infected with H. pylori SS1. A small cohort of human gastric mucosal biopsies was also examined. Treatment of AGS cells with H. pylori resulted in down-regulation of Id1 and Id3. Unexpectedly, expression of the main target of Id proteins, the basic helix-loop-helix transcription factor E2A, was also suppressed, with an associated decrease in E-box binding activity. In contrast, H. pylori induced the expression of the CDK inhibitor p21(WAF-1/cip1), and the homeobox transcription factor, Cdx2, an early marker of intestinal metaplasia of the stomach epithelium. Gastric epithelium from H. pylori-infected gerbils demonstrated similar changes, with decreased Id2, Id3 and E2A, and elevated p21(WAF-1/cip1) expression. In human gastric epithelium also, H. pylori infection was associated with reduced Id and E2A expression. In conclusion, H. pylori alters the expression of Id proteins, in vitro and in vivo; it is hypothesised that these changes contribute to H. pylori-associated pathologies. PMID- 16473540 TI - Candida albicans isolates with different genomic backgrounds display a differential response to macrophage infection. AB - Few human pathogens possess the ability exhibited by Candida albicans to colonize and cause symptomatic infections at different body sites. The host immune system is the major factor determining whether this opportunistic yeast behaves as a commensal or as a pathogen, since C. albicans strains appear capable of expressing similar virulence factors in response to specific body-district cues. This report provides evidence showing that C. albicans isolates with diverse genomic backgrounds (b and c karyotypes) differently modulate their pathogenic potential when assayed in cocultures with human monocytic derived macrophages (THP-1 cells). Striking differences were observed in the ability to undergo bud hypha transition, a relevant C. albicans virulence factor, between b and c karyotypes (P<0.0001) upon their internalization by macrophages. All c types were able to develop hyphal forms, resist intracellular killing, replicate, and escape from macrophages. The b type isolates, which were shown to be more efficiently ingested by THP-1 cells than the c type strains (P=0.013), were susceptible to intracellular killing and predominantly found as blastoconidia inside macrophages. Despite their different intracellular disposition, both b and c type isolates were equally able to undergo morphogenesis and to express NRG1 and HWP1 genes, markers of the bud-hypha transition program, during in vitro propagation. Since macrophages play a critical role in the host resistance to C. albicans, the different response of b and c isolates to macrophage infection suggests that the c type strains are better suited to behave as a more virulent strain cluster. PMID- 16473542 TI - Nomenclature and functions of RNA-directed RNA polymerases. AB - There is little relationship between eukaryotic RNA-directed RNA polymerases (RDRs), viral RNA-dependent RNA polymerases (RdRps) and DNA-dependent RNA polymerases, indicating that RDRs evolved as an independent class of enzymes early in evolution. In fungi, plants and several animal systems, RDRs play a key role in RNA-mediated gene silencing [post-transcriptional gene silencing (PTGS) in plants and RNA interference (RNAi) in non-plants] and are indispensable for heterochromatin formation, at least, in Schizosaccharomyces pombe and plants. Recent findings indicate that PTGS, RNAi and heterochromatin formation not only function as host defence mechanisms against invading nucleic acids but are also involved in natural gene regulation. RDRs are required for these processes, initiating a broad interest in this enzyme class. PMID- 16473541 TI - Does kinetics play a relevant role for intracellular staining? PMID- 16473544 TI - Do we need a new hypothesis to explain plant VOC emissions? PMID- 16473546 TI - CP43-like chlorophyll binding proteins: structural and evolutionary implications. AB - CP43, encoded by the psbC gene, is a chlorophyll (Chl)-binding protein of Photosystem II (PSII), the water-splitting and oxygen-evolving enzyme of photosynthesis. CP47, encoded by psbB, a Chl-binding protein of PSII, is closely related to CP43. The Chl-binding six transmembrane helical unit typified by CP43, is also structurally related to the N-terminal domains of the PsaA and PsaB proteins of Photosystem I (PSI) as well as to the family of light-harvesting proteins encoded by cyanobacterial isiA genes and prochlorophyte pcb genes. Here we use recent structural information derived for PSII and PSI to review similarities and differences between the various members of the CP43-like class of light-harvesting proteins, exploring both functional and evolutionary implications. PMID- 16473545 TI - Negative regulation of defence and stress genes by EAR-motif-containing repressors. AB - Although positive control or activation mechanism(s) involved in plant defence- and stress-related gene expression is relatively well studied, little is known about what keeps defensive armoury under control when not needed. Recent reports suggest that transcriptional repression of gene expression by EAR-motif containing repressor proteins plays a key role in modulating plant defence and stress responses. PMID- 16473547 TI - A clinical evaluation of Systane. AB - BACKGROUND: Systane is a new artificial tear solution containing the pH dependent gelling polymer hydroxypropyl-Guar (HP-Guar). We evaluated Systane in an open trial. METHODS: Patients complaining of dry feeling eyes were eligible to enroll, with inclusion criteria based on the presence of corneal staining with sodium fluorescein (NaFl) and participants' desire to use ocular lubricants. Six symptoms of ocular irritation were rated on a four-point scale. Conjunctival injection, conjunctival and corneal fluorescein staining were graded and tear break-up time (TBUT) and tear meniscus height were measured. Subjects were instructed to instill one drop of Systane into each eye four times a day for approximately four weeks. Ocular symptoms and signs were then reassessed and subjects were asked to rate eight statements concerning their satisfaction with the product. RESULTS: A total of 38 subjects were enrolled with 32 completing the follow-up visit. The following variables improved significantly at follow-up: all ocular irritation symptom scores, conjunctival injection, total conjunctival and total corneal staining scores, and TBUT. Tear meniscus height showed no significant change. Systane proved effective in reducing the symptoms of dry eye; however induced blur appeared to be a complication for many participants. CONCLUSIONS: The study represents a community based evaluation of the benefits of a novel ocular lubricant in optometric practice. The results suggest that the gelling properties of HP-Guar may be influential in promoting ocular surface recovery through improved ocular surface retention, however further randomised controlled trials are needed to confirm this. The extent and significance of induced blur also needs further investigation. PMID- 16473548 TI - Patient-directed intelligent and interactive computer medical history-gathering systems: a utility and feasibility study in the emergency department. AB - INTRODUCTION: Patients can be used as a resource to enter their own pertinent medical information. This study will evaluate the feasibility of an intelligent computer medical history-taking device directed at patients in the emergency department (ED). METHODS: Two of the authors (MB, RE) developed an expert system that can take patient-directed medical histories. Patients interacted with the computer in the ED waiting room while it gathered a medical history based on chief complaint (CC). A survey was completed post history. A sub-study assessed the computer's ability to take an adequate history for an index CC. We compared the computer and emergency physician histories for the presence or absence of important historical elements. RESULTS: Sixty-seven patients used the interactive computer system. The mean time to complete the history was 5 min and 32s +/- 1 min and 21s. The patient response rate was 97%. Over 83% felt that the computer was very easy to use and over 92% would very much use the computer again. A total of 15 patients with abdominal pain (index CC) were evaluated for the sub-study. The computer history asked 90+/-7%, and the emergency physician asked 55+/-18%, of the important historical elements. These groups were statistically different with a p-value of <0.00001. CONCLUSION: This feasibility study has shown that the computer history-taking device is well accepted by patients and that such a system can be integrated into the normal process of patient triage without delaying patient care. Such a system can serve as an initial mode for documentation and data acquisition directly from the patient. PMID- 16473549 TI - The usefulness of a multidisciplinary educational programme after breast cancer surgery: a prospective and comparative study. AB - The aim of the study was to compare and evaluate a multidisciplinary educational programme with traditional follow-up visits to a physician after breast cancer surgery in terms of well-being, aspects of self-care and coping ability 1 year after diagnosis. A reduction in the intensity of follow-up after breast cancer surgery is recommended. New follow-up models are being debated and could be of interest. The study design was non-randomised and comparative. Ninety-six consecutively selected women with newly diagnosed breast cancer, classified as stage I or stage II, participated in either a multidisciplinary educational programme (n=50), or traditional follow-up by a physician (n=46). Three questionnaires were used: Functional Assessment of Cancer Therapy-General (FACT G), a study specific questionnaire regarding self-care aspects (SCA) and Sense of Coherence (SOC). With the exception of physical well-being at baseline there was no significant difference between the groups. The women in the multidisciplinary educational programme increased their physical and functional well-being (P<0.01). The women in traditional follow-up by a physician increased their functional well-being while social/family well-being (P<0.01) decreased over time. There was a statistically significant difference in SOC (P<0.001) in the traditional follow-up by a physician between baseline (mean=74.4, SD=12.4) and the 1-year follow up (mean=67.7, SD=11.4). Thus, women in the traditional follow up by a physician scored lower in the area of SOC 1 year after diagnosis. A multidisciplinary educational programme may be an alternative to traditional follow-up by a physician after breast cancer surgery, but more research is needed about the financial benefits and effectiveness of such a programme. PMID- 16473550 TI - Spinal muscular atrophy: the RNP connection. AB - Degenerated motor neurons in the spinal cord are the pathological hallmark of spinal muscular atrophy (SMA). SMA is caused by mutations in the ubiquitously expressed survival motor neuron 1 (SMN1) gene, which lead to reduced levels of functional SMN protein. Many different functions have been assigned to SMN, including assembly of ribonucleoproteins (RNPs), splicing, transcription and axonal mRNA transport. Recently, tissue from SMA patients and animal models has been used to determine which function of SMN is affected in SMA patients. A surprising picture has emerged: the impaired assembly of RNP subunits of the spliceosome seems to be responsible for SMA pathogenesis. Here, we present a model of how this defect might cause motor-neuron degeneration and consider potential therapies. PMID- 16473551 TI - Inhibition of in vitro gamete adhesion and in vivo fertility in mice by immunization with a synthetic peptide. AB - PROBLEM: To evaluate the contraceptive ability of a synthetic peptide in in vitro and in vivo fertility in the mouse. METHOD OF STUDY: A synthetic peptide segment: GELRERAPGQGTNG (SP) was used to immunize female B6CF1 (C57BL/6 x BALB/c) mice. A peptide with an amino acid sequence QQPLSIQQHERG (p2control) was used as control. Anti-SP and anti-p2control antisera were used to evaluate sperm function inhibition in vitro. Fertility of immunized mice was determined by microscopic evaluation of the number and state of preimplantation embryos (8-16 cell stage). RESULTS: In the mouse, anti-SP antisera recognized surface antigens in the acrosome region of mature and capacitated sperm. Anti-SP antisera inhibited in vitro sperm binding to zona pellucida. In vivo, immune response against SP in Freund's adjuvant resulted in a significant increase (P < 0.01) in the number of dead embryos and eggs (a mean of 66%, in contrast with < 25% in control mice). Fertility inhibition in vivo and in vitro was not observed when the p2control peptide was used in the immunizations. CONCLUSIONS: These results would suggest that the SP sequence is involved in gamete adhesion, and an antifertility vaccine against the SP peptide segment could be feasible. PMID- 16473552 TI - A chimeric human-cat Fcgamma-Fel d1 fusion protein inhibits systemic, pulmonary, and cutaneous allergic reactivity to intratracheal challenge in mice sensitized to Fel d1, the major cat allergen. AB - Co-aggregation of FcepsilonRI with FcgammaRIIb can block FcepsilonRI-mediated reactivity and Fc gamma:allergen chimeric proteins, by co-crosslinking FcgammaRIIb to allergen-specific IgE bound to the FcepsilonRI can block allergen specific reactivity. We evaluated whether a human cat chimeric fusion protein (GFD) composed of part of the human Ig G1 Fc fused to the major cat allergen (Fel d1) would function as allergen immunotherapy while not inducing acute allergic reactivity in mice sensitized to Fel d1. Injection of GFD 6 h prior to Fel d1 challenge acutely blocked systemic and skin reactivity to Fel d1 challenge while mice given subcutaneous immunotherapy with GFD at days 37, 38, and 39 showed inhibition of systemic, lung, and cutaneous reactivity to Fel d1 2 weeks later. GFD immunotherapy did not induce systemic reactivity. Overall, the Fcgamma-Fel d1 chimeric fusion protein blocked Fel d1-induced IgE-mediated reactivity but did not induce in vivo mediator release on its own; suggesting that this approach using allergen combined with Fc gamma1 so as to achieve inhibitory signaling may provide an enhanced form of allergen immunotherapy. PMID- 16473554 TI - Removal of empty capsids from type 1 adeno-associated virus vector stocks by anion-exchange chromatography potentiates transgene expression. AB - Production of recombinant adeno-associated virus (rAAV) results in substantial quantities of empty capsids or virus-like particles (VLPs), virus protein shells without the vector genome. The contaminating VLPs would interfere with transduction by competing for cell-surface receptors and, when administered in vivo, contribute to antigen load, which may elicit a stronger immune response. Density-gradient ultracentrifugation provides a means to separate VLPs from rAAV particles, but is not feasible for large-scale preparations of vectors. Since the compositions of the VLP and vector differ by the single-stranded DNA genome, we hypothesized that the isoelectric point of the vector may differ from that of the VLP. In an attempt to separate type 1 rAAV particles from VLPs by ion-exchange chromatography, we tested a number of buffer systems and found that trimethylammonium sulfate, or [(CH3)4N]2SO4, effectively separated rAAV1 particles from VLPs. The rAAV1-GFP chromatographically separated from VLPs induced stronger GFP expression in HEK293 cells than rAAV1-GFP contaminated with VLPs. The transduction of mouse muscles with rAAV1-SEAP (secreted form of alkaline phosphatase) isolated from VLPs also showed higher serum SEAP levels than rAAV1-SEAP with VLPs. These results suggest that chromatographic separation of rAAV1 from empty capsids increased the efficacy of rAAV1. PMID- 16473553 TI - Constitutive and activation-inducible cyclooxygenase-2 expression enhances survival of chronic lymphocytic leukemia B cells. AB - We recently reported that activated normal human B lymphocytes express Cox-2. These findings prompted us to evaluate whether human B-CLL cells express Cox-2 and synthesize prostaglandins. In contrast to naive human B cells, B-CLL cells constitutively expressed Cox-2 protein and produced PGE2, PGF2alpha, and TXA2. Elevated Cox-2 expression was seen in a subgroup of B-CLL cells that exhibit poor prognostic factors, including unmutated variable heavy chain status and increased CD38 expression. Furthermore, stimulation of B-CLL cells with CD40 ligand plus TNFalpha increased Cox-2 levels. The role of Cox-2 in promoting B-CLL survival was investigated using nonselective and selective Cox-2 inhibitors. Significant reductions in B-CLL survival occurred following Cox-2 inhibition. These new findings support that constitutive Cox-2 expression in B-CLL cells promotes their survival and possibly their expansion in vivo. It will therefore be important to evaluate drugs that inhibit Cox-2 as potential therapeutic agents in B-CLL in vivo. PMID- 16473555 TI - Menhaden fish oil improves spatial memory in rat pups following recurrent pentylenetetrazole-induced seizures. AB - This study investigated the effects of two supplementary dietary oils (fish oil and corn oil) as parts of isocaloric/isoproteic diets on growth, brain fatty acid composition, and behavior in rat pups with recurrent seizures. Recurrent seizures were induced by injecting rat pups with pentylenetetrazole (PTZ) between P10 (10 days of age) and P14. Either menhaden fish oil (FO) or corn oil (CO) was given as supplemental dietary oil throughout the experiment from P3 to P40. We assessed the effects of the two supplemental dietary oils on spatial memory, histomorphology, and fatty acid composition of brain tissue at the end of the study on P40. Rats that received dietary FO performed significantly better in the Morris water maze, a test used to examine spatial performance in rats; the FO group had significantly shorter escape latencies (P=0.041) during the escape test. Compared with the CO group, the FO group stayed a longer time (P=0.015) and swam a longer distance (P=0.033) in the target quadrant in the spatial probe test. The FO group had significantly higher brain docosahexaenoic acid (P0.01) and significantly lower brain C20:3 n-6 and C20:4 n-6 (P<0.01 and P=0.031) levels compared with the CO group, but the two groups did not differ significantly with respect to neuronal cell loss in the histomorphology study. This study demonstrated that, compared with CO, FO is better in improving spatial memory in rats following recurrent PTZ-induced seizures. PMID- 16473556 TI - Hypersexuality in two patients with epilepsy treated with lamotrigine. AB - PURPOSE: Lamotrigine (LTG) is a novel anticonvulsant drug that exerts an antiepileptic effect by decreasing glutamate release through inhibition of voltage-sensitive sodium channels. LTG has no effect on serum levels of most female reproductive hormones, but its effect on male reproductive hormones still remains unclear. Improvement in sexual function after LTG treatment has been reported, and could have been caused by reduction of seizures, inhibition of focal discharges, or an unknown effect of LTG on reproductive hormones and protein levels. CASES: Two male patients exhibited acute hypersexuality while taking lamotrigine as add-on therapy: one patient on carbamazepine and one on oxcarbazepine. Neither prior history of psychiatric illness nor brain damage predisposed them to such a response to treatment, and in both patients, the hypersexuality was not a part of hypomania or a more diffuse psychiatric disturbance. In the first case, sexual hyperactivity resolved after discontinuation of LTG therapy without any concomitant treatment. In the second case, a reduction in the dose of LTG decreased the intensity of the hypersexuality and contributed to the patient's increased satisfaction with his sex life. CONCLUSIONS: Lamotrigine may cause drug-related hypersexuality by an unclear underlying mechanism. PMID- 16473557 TI - The impact of levetiracetam on challenging behavior. AB - An observational longitudinal design was employed to evaluate whether treatment with the antiepileptic drug levetiracetam (LEV) adversely impacts behavior in people with intellectual disabilities and/or acquired brain damage. Thirty-five adults were assessed once off the drug and once when on LEV therapy, with a 2 month interval between assessments. Behaviors were rated using an adaptation of the Yale-Brown Obsessive Compulsive Scale and the Challenging Behaviour Scale. Challenging behaviors were rated as more frequent and severe when individuals were taking LEV. Behavioral worsening was not related to better seizure control or increased levels of engagement in activities. Families and professionals need to be aware of the potential reversible adverse effects of this drug. PMID- 16473558 TI - Every-12-hour administration of extended-release divalproex in patients with epilepsy: impact on plasma valproic acid concentrations. AB - Extended-release divalproex sodium (divalproex-ER) biopharmaceutics after every 12-hour (q12h) administration was compared with that of once-daily divalproex-ER and conventional divalproex given every 6 hours (q6h) in a multiple-dose (14 day), randomized, three-period crossover design study in 24 patients with epilepsy concomitantly receiving enzyme-inducing antiepileptic medication(s). Plasma valproic acid (VPA) minimum concentration (Cmin) for divalproex-ER q12h was higher than the once-daily divalproex-ER Cmin (P=0.043). Once-daily divalproex-ER Cmin values were not different from those for divalproex q6h, suggesting that adequate trough steady-state concentrations are maintained with once daily dosing, despite enzyme-inducing comedication. The degree of peak trough fluctuation (DFL, calculated as (Cmax-Cmin)/Cavg) in VPA concentration was less with both q12h (35.2% less) and once-daily (16.9% less) divalproex-ER regimens compared with q6h divalproex (P0.024). The DFL for divalproex-ER dosed as a q12h regimen was 22% less than that for once-daily divalproex-ER (P=0.02). The DFL in VPA concentration with divalproex-ER can be minimized with once-daily administration and more so with q12h administration, compared with conventional enteric-coated divalproex taken q6h. PMID- 16473559 TI - Traumatic intracranial hemorrhage in patients with seizures: descriptive characteristics. AB - We reviewed the records of all patients with recurrent seizures and severe head injury-induced traumatic intracranial hemorrhage (TIH) between 1989 and 2003 in three Israeli medical centers. We identified 52 cases (44 males, mean age=43+/-19 years, range=8-84; 8 females; mean age=74+/-12 years, range=48-85). Twenty-seven (52%) had additional known risk factors for TIH, e.g., older age, alcohol abuse, and anticoagulant use. All five children and adolescents had mental retardation. Approximately one-half of patients with seizures and TIH have additional risk factors for TIH. Non-mentally retarded children and adolescents with seizures are probably at low risk of developing TIH. Women less than 70 years old with seizures are much less prone to TIH than men. In young "otherwise healthy" patients with epilepsy, suboptimal treatment seems to be an important factor in the occurrence of TIH. PMID- 16473560 TI - Development of methods for extraction and in vitro quantification of estrogenic and androgenic activity of wastewater samples. AB - Chemicals released into the environment by anthropogenic activities have been linked to estrogenic or androgenic effects in exposed wildlife, and there is a need to develop and validate rapid and cost-effective methods to quantify the total estrogenic and androgenic activity of environmental water samples. In this study, estrogen receptors (ER) were isolated from sheep (Ovis aries) uteri and rainbow trout (Oncorhynchus mykiss) livers and androgen receptors (AR) were isolated from rainbow trout brains. The isolated receptors were used in competitive receptor binding assays to test the affinity of known estrogenic and androgenic chemicals for the receptor binding site, and results were compared with literature values for the rat uterine ER binding assay and the E-Screen. The relative binding affinities of the tested compounds to ER from different species were similar, and binding to the ER was a more responsive endpoint than the cellular effect measured in the E-Screen. Using the sheep ER binding assay in combination with solid-phase extraction, the estrogenic activity in a raw sewage sample from a municipal treatment plant in Brisbane (Queensland, Australia) was measured at 51-73 ng/L estradiol equivalents (EEq). PMID- 16473561 TI - A normal mucin-binding lectin from the sponge Craniella australiensis. AB - A lectin, Craniella australiensis (CAL), was isolated from sponge C. australiensis by ion-exchange on DEAE-Sephacel and purified by gel filtration on Sephadex G-150 and HPLC on DEAE-5PW. The purified lectin was a trimeric protein as revealed by SDS-PAGE and MALDI-TOF analysis. SDS-PAGE showed that the CAL protein had a molecular mass of 54 kDa, and consisted of three 18 kDa subunits. Gel filtration of purified lectin on Sephadex G-200 indicates that it exists as a 54 kDa protein in its native state. The amino acid composition was rich in Thr and Glx. CAL was found to agglutinate native and trypsinized human A, B erythrocytes, and agglutinate native erythrocytes of mouse, sheep, rabbit and chicken, and trypsinized erythrocytes of sheep and rabbit. The hemagglutination activity was inhibited by glycoproteins such as PSM and asialo-PSM, but not by any of the monosaccharides tested. The activity was stable between 20 and 70 degrees C. Significant CAL activity was observed between pH 5 and 8. The lectin reaction is independent of the presence of divalent cations Ca2+ and Mg2+. The sequence of N-terminal residues of CAL was determined as TSSCQSIVVE. The lectin showed a potent mitogenic response towards BALB/c splenocytes. PMID- 16473562 TI - Signalling the state of the digestive tract. AB - The gastrointestinal tract has a rich sensory innervation. Extrinsic afferents in vagal, splanchnic and pelvic nerves project to the CNS where gut reflex function is coordinated and integrated with behavioural responses (e.g. regulation of food intake) and mediate sensations. The afferent information conveyed by vagal and spinal mechanosensitive afferents can be very different. Vagal afferents have low thresholds of activation and reach maximal responses within physiological levels of distension. In contrast, spinal afferents, although many have corresponding thresholds for activation, are able to respond beyond the physiological range and encode both physiological and noxious levels of stimulation. However, mechanosensitivity is not fixed but can be influenced by a wide range of chemical mediators released as a consequence of ischemia, injury and inflammation. Indeed, previously mechanical insensitive afferents can develop mechanosensitivity during inflammation and a variety of chemical mediators are implicated in this sensitisation process. Chemosensitivity is also a property of vagal mucosal afferents that detect the chemical milieu for chemicals absorbed across the epithelium or released from enteroendocrine cells that are strategically positioned to "taste" luminal contents. Thus, there exists a complex interplay between immunomodulators, neurotransmitters and neuroendocrine factors that underlie gastrointestinal sensing mechanisms and enable orchestration of appropriate host responses. PMID- 16473563 TI - Phylogenetic and evolutionary aspects of Paracoccidioides brasiliensis reveal a long coexistence with animal hosts that explain several biological features of the pathogen. AB - The habitat of the mycelial saprobic form of Paracoccidioides brasiliensis, which produces the infectious propagula, has not been determined and has proven difficult for mycologists to describe. The fungus has been rarely isolated from the environment, the disease has a prolonged latency period and no outbreaks have been reported. These facts have precluded the adoption of preventive measures to avoid infection. The confirmation of natural infections in nine-banded armadillos (Dasypus novemcinctus) with P. brasiliensis, in high frequency and wide geographic distribution, has opened new avenues for the study and understanding of its ecology. Armadillos belong to the order Xenarthra, which has existed in South America ever since the Paleocene Era (65 million years ago), when the South American subcontinent was still a detached land, before the consolidation of what is now known as the American continent. On the other hand, strong molecular evidence suggests that P. brasiliensis and other dimorphic pathogenic fungi--such as Blastomyces dermatitidis, Coccidioides immitis and Histoplasma capsulatum- belong to the family Onygenaceae sensu lato (order Onygenales, Ascomycota), which appeared around 150 million years ago. P. brasiliensis ecology and relation to its human host are probably linked to the fungal evolutionary past, especially its long coexistence with and adaptation to animal hosts other than Homo sapiens, of earlier origin. Instead of being a blind alley, the meaning of parasitism for dimorphic pathogenic fungi should be considered as an open two-way avenue, in which the fungus may return to the environment, therefore contributing to preserve its teleomorphic (sexual) and anamorphic (asexual) forms in a defined and protected natural habitat. PMID- 16473564 TI - HIV-1 subtypes and recombinants in the Republic of Congo. AB - To document the actual genetic diversity of HIV-1 strains in the Republic of Congo, 114 HIV-1 positives persons were sampled in 2003 and 2004 after their informed consent. They were attending the teaching hospital, the reference health center in Makelekele, Brazzaville and the regional hospital centers in Pointe Noire, Gamboma and Ouesso. A total of 104 samples were genetically characterized by direct sequencing of the p24 gag region and 80 were also subtyped in the V3-V5 env region. The genetic subtype distribution of the Congolese strains showed the predominance of subtype A (36.5% and 32.5% in gag and env, respectively) and G (30.8% and 21.25%), whereas subtype D strains represented 12.5% and 15%. Subtypes C, F, H, J, K and the CRFs-01, -02, -05 -06, and also the recently characterized CRF18 were seen at lower rates. Finally, 4.8% (gag) and 6.25% (env) of the strains could not be classified. Moreover, a high intra-subtype diversity was observed in our study. Among 70 strains which have been characterized in the two genomic regions, 14 (20%) appeared to be unique recombinants. These data show a high genetic variability in the Republic of Congo, where all the subtypes have been documented together with certain subsubtypes and several CRFs. PMID- 16473565 TI - Nitro radical anion formation from nitrofuryl substituted 1,4-dihydropyridine derivatives in mixed and non-aqueous media. AB - Three new nitrofuryl substituted 1,4-dihydropyridine derivatives were electrochemically tested in the scope of newly found compounds useful as chemotherapeutic alternative to the Chagas' disease. All the compounds were capable to produce nitro radical anions sufficiently stabilized in the time window of the cyclic voltammetric experiment. In order to quantify the stability of the nitro radical anion we have calculated the decay constant, k2. Furthermore, from the voltammetric results, some parameters of biological significance as E7(1) (indicative of in vivo nitro radical anion formation) and KO2 (thermodynamic indicator of oxygen redox cycling) have been calculated. From the comparison of E7(1), KO2 and k2 values between the studied nitrofuryl 1,4-DHP derivatives and well-known current drugs an auspicious activity for one of the studied compounds i.e. FDHP2, can be expected. PMID- 16473566 TI - Translesion DNA synthesis across non-DNA segments in cultured human cells. AB - DNA lesions that have escaped DNA repair are tolerated via translesion DNA synthesis (TLS), carried out by specialized error-prone DNA polymerases. To evaluate the robustness of the TLS system in human cells, we examined its ability to cope with foreign non-DNA stretches of 3 or 12 methylene residues, using a gap lesion plasmid assay system. We found that both the trimethylene and dodecamethylene inserts were bypassed with significant efficiencies in human cells, using both misinsertion and misalignment mechanisms. TLS across these non DNA segments was aphidicolin-sensitive, and did not require poleta. In vitro primer extension assays showed that purified poleta, polkappa and poliota were each capable of inserting each of the four nucleotides opposite the trimethylene chain, but only poleta and polkappa could fully bypass it. Poleta and poliota, but not polkappa, could also insert each of the four nucleotides opposite the dodecamethylene chain, but all three polymerases were severely blocked by this lesion. The ability of TLS polymerases to insert nucleotides opposite a hydrocarbon chain, despite the lack of any similarity to DNA, suggests that they may act via a mode of transient and local template-independent polymerase activity, and highlights the robustness of the TLS system in human cells. PMID- 16473567 TI - A practical approach to optimization and validation of a HPLC assay for analysis of polyribosyl-ribitol phosphate in complex combination vaccines. AB - The use of multi-factor statistical experimental design methodology minimized the vaccine material and laboratory resources required for optimization and validation of an HPLC assay for quantitation of depolymerized and total PRP. Components of the assay selected for optimization were adjuvant dissolution, ultracentrifuge conditions including ultracentrifuge model, sample diluent, mobile phase and column oven temperature. Previous experience has shown these components of the assay to be most troublesome and therefore required optimization prior to validation. Specificity, linearity, precision, accuracy and ruggedness were confirmed through a validation of the optimized assay. The validation also established the assay to be stability indicating, by showing that changes to the integrity of the PRP-OMPC conjugate could be detected. PMID- 16473569 TI - Methodology and assessments: the tools of the trade. PMID- 16473568 TI - Cross-sectional reference data for phalangeal quantitative ultrasound from early childhood to young-adulthood according to gender, age, skeletal growth, and pubertal development. AB - Bone mineral status by phalangeal quantitative ultrasound (QUS, DBM Sonic, IGEA, Carpi, Modena, Italy) was examined in 3044 (1513 males and 1531 females) healthy subjects, aged 2-21 years. The aim of the study was to provide a reference database for phalangeal QUS parameters, amplitude-dependent speed of sound (AD SoS) and bone transmission time (BTT), both expressed as centiles and Z score, according to gender, age, height, weight, body mass index (BMI), and pubertal stage to be used for estimating bone mineral status in patients with disorders of growth or of bone and mineral metabolism. In both sexes, AD-SoS and BTT increased significantly (P<0.0001) according to all the anthropometric variables. Females showed higher values than males in the age groups 9-14 for AD-SoS (P<0.04 P<0.0001) and in the age groups 11-13 for BTT (P<0.02). Males had higher BTT values than females in the age groups 6-8 and 15-21 (P<0.04-P<0.0001). AD-SoS was higher (P<0.02-P<0.0001) in females than in males at pubertal stages 2, 3, and 4, but it was higher (P=0.001) in males compared with females at pubertal stage 5. BTT was higher in males than females at pubertal stages 1 (P<0.0001), 2 (P<0.01), and 5 (P<0.0001). In both sexes, AD-SoS and BTT were significantly correlated between them (r=0.92, P<0.0001) and with all the anthropometric variables (r=0.53 r=0.85, P<0.0001). Age, weight, BMI, and pubertal stage were independent predictors of AD-SoS in males; age and pubertal stage were independent predictors of AD-SoS in females. In both sexes, height and pubertal stage, and also age only in females, were independent predictors of BTT. In conclusion, our data show that gender, age, height, and timing of sexual maturation are main determinants of bone structure and geometry, and that both these two processes may be captured by phalangeal QUS. It may be a useful tool to assess bone mineral status from early childhood to young-adulthood with a very small confounding effect related to bone sizes and without exposing the subjects to a source of radiation. PMID- 16473570 TI - Upper airway resistance syndrome: a long-term outcome study. AB - This prospective study aimed to assess symptomatic evolution of patients diagnosed with Upper Airway Resistance Syndrome (UARS) four and half years after the initial UARS diagnosis. For this purpose, 138 UARS patients were contacted by mail between 43 and 69 months after the initial evaluation; 105 responded to the letter and 94 patients accepted to undergo new clinical and polysomnographic evaluations. Initial and follow-up polysomnographic recordings were scored using the same criteria. RESULTS: Of the 94 patients who completed the follow-up examination, none of them were using nasal CPAP. It was related to refusal by insurance providers to provide equipment based on initial apnea-hypopnea index (AHI) in 90/94 subjects. Percentage of patients with sleep related-complaints significantly increased over the four and half year period: daytime fatigue, insomnia and depressive mood increased by 12 to 20 times. Reports of sleep maintenance sleep onset insomnia and depressive mood was significantly increased. Hypnotic, antidepressant and stimulant prescription increased from initial to follow-up visit (from 11.7% to 61.7%; from 3.2% to 25.5% and from 0% to 9.6%, respectively) with antidepressant given as much for sleep disturbance as mood disorder. The polysomnography results at follow-up showed that 5 subjects had AHI compatible with Obstructive Sleep Apnea Syndrome (OSAS) but overall, respiratory disturbance index had no significant change. Total sleep time was significantly reduced compared to initial visit. CONCLUSIONS: Many UARS patients remained untreated following initial evaluation. Worsening of symptoms of insomnia, fatigue and depressive mood were seen with absence of treatment of UARS. PMID- 16473573 TI - Use of search algorithms to define specificity in Rab GTPase domain function. AB - The continuing explosion of sequencing data has inspired a corresponding effort in the annotation and classification of protein families. Within a particular protein family, however, individual members may have distinct functions, although they share a common fold and broadly defined physiological role. Rab GTPases are the largest subfamily of the Ras superfamily, yet from early in their discovery, it was apparent that each Rab protein has a unique subcellular localization and regulates a particular stage(s) membrane traffic. To gain insight into the contribution of individual residues to unique protein functions a general strategy is outlined. This method should allow the cell and molecular biologist with no specialist expertise to implement an algorithm that makes use of a combination of experimental and phylogenetic data. The algorithm is applicable to the analysis of any protein domain and here is illustrated with the analysis of residues contributing to the individual functions of a pair of Rab GTPases. PMID- 16473572 TI - Exploring trafficking GTPase function by mRNA expression profiling: use of the SymAtlas web-application and the Membrome datasets. AB - Despite complete sequencing of the human and mouse genomes, functional annotation of novel gene function still remains a major challenge in mammalian biology. Emerging strategies to help elucidate unknown gene function include the analysis of tissue-specific patterns of mRNA expression. A recent study investigated the steady-state mRNA expression profiling of the vast majority of protein-encoding human and mouse genes across a panel of 79 human and 61 mouse nonredundant tissues. The microarray data from this study constitutes the Genomics Institute of Novartis Foundation (GNF) Human and Mouse Gene Atlases and is publicly available for exploration through the SymAtlas web-application (http://symatlas.gnf.org/). We have recently reported the use of these data and hierarchical clustering algorithms to generate a global overview of the distribution of Rabs, SNAREs, and coat machinery components, as well as their respective adaptors, effectors, and regulators. This systems biology approach led us to propose Rab-centric protein activity hubs as a framework for an integrated coding system, the membrome network, which orchestrates the dynamics of specialized membrane architecture of differentiated cells. Here, we describe the use of the SymAtlas web-application and the Membrome datasets to help explore trafficking GTPase function. The human and mouse membrome datasets are available through the Membrome homepage (http://www.membrome.org/) and correspond to subsets of the SymAtlas content restricted to known membrane trafficking components. Considering the fragmentary nature of the current reductionist approaches in elucidating trafficking component functions, the membrome datasets provide a more focused systems biology perspective that not only complements our current understanding of transport in complex tissues but also provides an integrated perspective of Rab activity in controlling membrane architecture. PMID- 16473574 TI - Application of phylogenetic algorithms to assess Rab functional relationships. AB - Researchers looking to solve biological problems have access to enormous amounts of sequence information and the desktop computational infrastructure to personally interrogate and analyze large datasets. Many powerful bioinformatics tools are available online; however, this discourages the customized analysis of data that is necessary for the experimental scientist to make maximally effective use of the information. In addition, a customized environment facilitates the critical evaluation of bioinformatic methods. This chapter presents a protocol developed to aid in classification of subfamilies and subclasses of a superfamily using the personal desktop computer. The visual representation of the qualitative and quantitative results of data analyses is also considered. The examples are focused on Rab GTPases but are more widely applicable to the classification of any given protein family. PMID- 16473575 TI - Application of protein semisynthesis for the construction of functionalized posttranslationally modified rab GTPases. AB - Rab GTPases represent a family of key membrane traffic regulators in eukaryotic cells. To exert their function, Rab proteins must be modified with one or two geranylgeranyl moieties. This modification enables them to reversibly associate with intracellular membranes. In vivo the newly synthesized Rab proteins are recruited by Rab escort protein (REP) that presents them to the Rab geranylgeranyl transferase (Rab GGTase), which transfers one or two geranylgeranyl moieties to the C-terminal cysteines. Detailed understanding of the mechanism of prenylation reaction and subsequent membrane delivery of Rab proteins to the target membranes were hampered by lack of efficient technologies for the generation of preparative amounts of prenylated Rab GTPases. To circumvent this problem, we developed an approach that combines recombinant protein production, chemical synthesis of lipidated peptides with precisely designed and readily alterable structures, and a technique for peptide-to-protein ligation. Using this approach, we generated a number of semisynthetic prenylated Rab GTPases. Some of the proteins were also supplemented with fluorophores, which enabled us to develop a fluorescence-based in vitro prenylation assay. The approach described allows production of preparative amounts of prenylated GTPases, which was demonstrated by generation and crystallization of a monoprenylated YPT1:Rab GDI complex. PMID- 16473576 TI - Assay and functional properties of PrBP(PDEdelta), a prenyl-binding protein interacting with multiple partners. AB - A 17-kDa prenyl-binding protein, PrBP(PDEdelta), is highly conserved among various species from human to Caenorhabditis elegans. First identified as a putative regulatory delta subunit of the cyclic nucleotide phosphodiesterase (PDE6) purified from mammalian photoreceptor cells, PrBP(PDEdelta) has been hypothesized to reduce activation of PDE6 by the heterotrimeric G-protein, transducin, thereby desensitizing the photoresponse. However, recent work shows that PrBP(PDEdelta) interacts with numerous prenylated proteins at their farnesylated or geranylgeranylated C-termini, as well as with non-prenylated proteins. These polypeptides include small GTPases such as Rab13, Ras, Rap, and Rho6, as well as components involved in phototransduction (e.g., rod and cone PDE6, rod and cone opsin kinases). Expression of PrBP(PDEdelta) in tissues and organisms not expressing PDE6, the demonstration of multiple interacting partners with PrBP(PDEdelta), and its low abundance in rod outer segments all argue against it being a regulatory PDE6 subunit. This raises intriguing questions as to its physiological functions. In this chapter, we review the current status of PrBP(PDEdelta) and describe some of the assays used to determine these interactions in detail. In mammalian photoreceptors, the results are consistent with a role of PrBP(PDEdelta) in the transport of prenylated proteins from their site of synthesis in the inner segment to the outer segment where phototransduction occurs. PMID- 16473577 TI - Functional assays for the investigation of the role of Rab GTPase effectors in dense core granule release. AB - Rab3 and Rab27 GTPases control late events in the secretory pathway of mammalian cells including docking and fusion of secretory vesicles with the plasma membrane. The action of Rab3 and Rab27 on the exocytotic process is exerted through the activation of specific effectors. Several proteins with the capacity to interact in a GTP-dependent manner with Rab3 and Rab27 have been identified. However, for most of these potential Rab effectors a precise function in the secretory process has not yet been attributed. In this chapter we describe a series of approaches that can be applied to assess the properties of Rab3 and Rab27 effectors and their potential role in the regulation of dense core granule release. PMID- 16473578 TI - Analysis of Rab1 recruitment to vacuoles containing Legionella pneumophila. AB - Legionella pneumophila is an intracellular bacterial pathogen that utilizes a type IV secretion system to subvert the function of the Rab1 GTPase. Bacterial proteins translocated into host cells mediate the accumulation of the Rab1 protein on vacuoles containing Legionella pneumophila. Assays used to investigate recruitment of Rab1 by L. pneumophila are described. These assays can be used to determine host and bacterial factors required for L. pneumophila subversion of the host secretory pathway. PMID- 16473579 TI - Reconstitution of Rab4-dependent vesicle formation in vitro. AB - We have developed an in vitro assay to reconstitute the formation of endosomal recycling vesicles. To achieve specificity for endosomes as the donor organelle, cells are surface-biotinylated and allowed to endocytose for 10 min, after which the remaining surface-biotin is stripped off. The cells are then permeabilized and the cytosol washed away. Upon addition of exogenous cytosol and energy, sealed vesicles containing biotinylated recycling receptors are produced. Modification of the cytosol, for example, by immunodepletion or addition of purified proteins, allows the identification of proteins involved in vesicle formation. The results show that recycling is mediated by AP-1/clathrin-coated vesicles, requires Rab4, and is negatively regulated by rabaptin-5/rabex-5. PMID- 16473580 TI - Assay of Rab4-dependent trafficking on microtubules. AB - We present an in vitro method to measure how Rab4 and other regulatory proteins affect microtubule-based organelle motility. The protocols utilize small-volume, disposable "microchambers" designed for epifluorescence, confocal, or other microscope platforms and into which microtubules, organelles, and primary and fluorescent secondary antibodies are added. Our work has focused on the isolation and use of endocytic vesicles from rat liver, and we present these protocols. However, the techniques can be adapted for other organelles or cell types. Multiple fluorescent probes, rapid image capture, and immunofluorescence under non-fixation conditions allow for measurements of the location and intensity changes of endogenous proteins upon addition of ATP or upon addition of other proteins or regulatory factors. We review measurements of microtubule-based motility as well as measurements for protein localization and protein segregation in vitro. PMID- 16473581 TI - CD2AP, Rabip4, and Rabip4': analysis of interaction with Rab4a and regulation of endosomes morphology. AB - In this chapter, we describe various approaches that allow us to study interactions between the small GTPase Rab4a and its two effectors, Rabip4 and CD2AP. Two complementary approaches, one using the yeast two-hybrid system and the other using a GST pull-down assay, are described. We document the studies of the localization of these proteins by cellular fractionation. Finally, we develop cellular imaging techniques to study the morphology of vesicular structures containing Rab4a. We show that the coexpression of Rab4a with its effectors affects Rab4a-containing structures, giving a clear indication of their interaction in the mammalian cellular context. PMID- 16473582 TI - Visualization of Rab5 activity in living cells using FRET microscopy. AB - Rab5 is a member of the large family of small GTPases involved in membrane trafficking. Two genetically encoded sensors were developed to visualize Rab5 in its GTP-bound conformation in living cells. Rab5-binding fragments of Rabaptin5 or early endosomal antigen 1 (EEA.1) were fused to yellow fluorescent protein (YFP) and used in the fluorescent resonance energy transfer (FRET) assay together with Rab5-tagged cyan fluorescent protein (CFP). The presence of energy transfer between CFP-Rab5 and YFP-Rab5 binding fragments detected by sensitized FRET microscopy has validated the utility of these generated sensors to visualize the localization of GTP-bound Rab5. GTP-bound Rab5 was found in endosomes, often concentrated in distinct microdomains. Molecular architecture of the Rab5 microdomains was analyzed by three-chromophore FRET (3-FRET) microscopy, utilizing YFP, CFP, and monomeric red fluorescent proteins (mRFP.l). The results of the 3-FRET analysis suggest that GTP-bound Rab5 is capable of oligomerization and present in multiprotein complexes. PMID- 16473583 TI - Selection and application of recombinant antibodies as sensors of rab protein conformation. AB - The existence of a conformational switch of Rabs and other small GTPases involved in intracellular transport regulation has been known for many years. This switch is superimposed on the membrane association/dissociation cycle for most of these GTPases. While these processes are key features of the dynamics of intracellular transport events, surprisingly very few previous studies have focused on the dynamics of the GDP/GTP cycle of Rab proteins in time and space. The main reason for this is the lack of tools available to dynamically probe for Rab GTPases conformation switches and membrane association/dissociation, in particular in vivo. We recently reported the in vitro selection of conformation-specific recombinant antibodies specific to the GTP-bound conformation of Rab6 proteins. These antibodies were obtained in vitro by phage display, a rather simple, rapid, and cheap technique. We additionally showed that these conformation-specific antibodies can be expressed in living cells to follow endogenous Rab6 in its activated conformation in vivo. The same strategy could be used to study other conformation switching mechanisms and, in general, to study the switching between states that antibodies can distinguish (e.g., phosphorylation, ubiquitination). PMID- 16473585 TI - Use of Rab GTPases to study lipid trafficking in normal and sphingolipid storage disease fibroblasts. AB - We describe methods for studying lipid transport in normal and sphingolipid storage disease fibroblasts. These techniques include endocytic assays with fluorescent sphingolipid analogs, expression of dominant negative (DN) Rab GTPases, and methods of manipulating cholesterol levels in intact cells and isolated cell membranes. These methods should be useful in future studies of lipid trafficking in normal and disease cell types. PMID- 16473586 TI - Assay of Rab13 in regulating epithelial tight junction assembly. AB - Rab13 is recruited to tight junctions from a cytosolic pool after cell-cell contact formation. Tight junctions are intercellular junctions that separate apical from basolateral domains and are required for the establishment/maintenance of polarized transport in epithelial cells. They form selective barriers regulating the diffusion of ions and solutes between cells. They also maintain the cell surface asymmetry by forming a "fence" that prevents apical/basolateral diffusion of membrane proteins and lipids in the outer leaflet of the plasma membrane. We generate stable MDCK cell lines expressing inactive (T22N mutant) and constitutively active (Q67L mutant) Rab13 as GFP-Rab13 chimeras. Expression of GFP-Rab13Q67L delays the formation of electrically tight epithelial monolayers, induces the leakage of small nonionic tracers from the apical domain, and disrupts the tight junction fence diffusion barrier. It also alters the tight junction strand structure and delays the localization of the tight junction transmembrane protein, claudin1. In contrast, the inactive Rab13T22N mutant does not disrupt tight junction functions, tight junction strand architecture, or claudin1 localization. Here we describe a set of assays that allows us to investigate the role of Rab13 in modulating tight junction structure and function. PMID- 16473584 TI - Analysis of Rab protein function in neurotransmitter receptor trafficking at hippocampal synapses. AB - Members of the Rab family of small GTPases are essential regulators of intracellular membrane sorting. Nevertheless, very little is known about the role of these proteins in the membrane trafficking processes that operate at synapses, and specifically, at postsynaptic terminals. These events include the activity dependent exocytic and endocytic trafficking of AMPA-type glutamate receptors, which underlies long-lasting forms of synaptic plasticity such as long-term potentiation (LTP) and long-term depression (LTD). This chapter summarizes different experimental methods to address the role of Rab proteins in the trafficking of neurotransmitter receptors at postsynaptic terminals in the hippocampus. These techniques include immunogold electron microscopy to ultrastructurally localize endogenous Rab proteins at synapses, molecular biology methods to express recombinant Rab proteins in hippocampal slice cultures, electrophysiological techniques to evaluate the role of Rab proteins in synaptic transmission, and confocal fluorescence imaging to monitor receptor trafficking at dendrites and spines and its dependence on Rab proteins. PMID- 16473587 TI - Tyrosine phosphorylation of Rab proteins. AB - Tyrosine phosphorylation is a fundamental mechanism for regulating the functions of numerous proteins in eukaryotic cells. It has been known for some time that several members of the Rab GTPase family can undergo phosphorylation on serine or threonine residues, but the potential for tyrosine phosphorylation has been appreciated only recently, based on a single example-Rab24. Herein we describe a series of straightforward methods to facilitate an initial assessment of the potential for tyrosine phosphorylation of epitope-tagged Rab proteins transiently expressed in mammalian cells. The approach takes advantage of the availability of highly specific monoclonal antibodies against phosphotyrosine and specific chemical inhibitors for tyrosine kinases. We also describe the use of site directed mutagenesis to identify tyrosine residues that may be targets for phosphorylation, and we discuss the possible relevance of this modification for regulating Rab function. PMID- 16473588 TI - Assay of Rab25 function in ovarian and breast cancers. AB - There is a multitude of critical steps during the pathogenesis of cancer that allow cells to acquire the ability to escape from normal controls on cell growth, to avoid programmed cell death, and to become malignant. Here, we describe a molecular approach that can be broadly applied to identify drivers of genomic aberrations in cancer development. In the process, areas of genomic aberrations and genes that are dysregulated by genomic amplification are identified by array comparative genomic hybridization (CGH) and transcription profiling, respectively, with major emphasis on coordinating amplification at the CGH and RNA level and on correlation with patient's outcomes. Once candidate genes are identified, we perform functional genomics by manipulating levels in normal and tumor cells using RNAi or transfection, and assessing a battery of cellular functions including proliferation, anti-apoptosis, loss of contact inhibition, changes in cell signaling or transcriptional profiles, anchorage-independent growth, and in vivo tumor growth. We have successfully used this approach to identify the RAB25 gene that has been implicated in the progression and aggressiveness of ovarian and breast cancers. PMID- 16473589 TI - Functional analysis of Rab27a effector granuphilin in insulin exocytosis. AB - Granuphilin is specifically expressed on dense-core granules in a defined set of secretory cells such as insulin-producing pancreatic beta-cells. It preferentially binds the GTP-bound form of Rab27a and regulates the exocytosis of secretory granules. Furthermore, granuphilin directly interacts with syntaxin-la, the plasma-membrane-anchored SNARE protein, and with Munc18-1, a Sec1/Munc18 protein. We previously reported evidence that granuphilin mediates the docking of secretory granules onto the plasma membrane through these protein-protein interactions. This chapter details the methods and protocols we use to analyze the function of granuphilin with particular attention to the assays for detecting the expression, protein interactions, and effects on exocytosis of secretory granules in pancreatic beta-cells and their derivative cell lines. PMID- 16473591 TI - Fluorescent microscopy-based assays to study the role of Rab22a in clathrin independent endocytosis. AB - The endocytic and trafficking route followed by proteins that enter cells independently of clathrin is not well understood. In HeLa cells, a distinct endocytic and recycling pathway is followed for such clathrin-independent cargo proteins, and we have characterized the role of Rab proteins in this process. Here we describe cell-based flourescence assays to examine the effects of expression and depletion of Rab22 on endosomal morphology and endocytic recycling. PMID- 16473590 TI - Assays for functional properties of Rab34 in macropinosome formation. AB - We have shown that Rab34/Rah participates in the promotion of macropinosome formation. Here we describe procedures for the analyses of intracellular localization and some functional properties of Rab34. Rab34 lacks a consensus sequence of the fourth motif for GTP/GDP binding and GTPase activities. Indeed, GTPase assay shows that wild-type Rab34 has extremely weak GTPase activity in vitro. However, Rab34 exhibits appreciable GTPase activity in vivo probably due to the presence of specific GTPase-activating protein (GAP) activity in cells. Specific intracellular localization of Rab34 is easily detected by the expression of epitope-tagged or enhanced green fluorescent protein (EGFP)-tagged protein. It is colocalized with actin filaments to membrane ruffles and membranes of nascent macropinosomes, which are formed from the ruffles. By contrast, Rab5 is not associated with the ruffles or nascent macropinosomes but present in endosomes at later stages. The function of Rab34 in macropinosome formation is analyzed by the transfection of wild-type, constitutively active, and dominant-negative mutants of Rab34 in fibroblasts followed by treatment with platelet-derived growth factor (PDGF) or phorbol ester. These analyses indicate that Rab34 is required for efficient macropinosome formation. PMID- 16473592 TI - Purification and properties of Rab3 GEP (DENN/MADD). AB - Rab3A, a member of the Rab3 small GTP-binding protein (G protein) family, regulates Ca(2+)-dependent exocytosis of neurotransmitter. Rab3A cycles between the GDP-bound inactive and GTP-bound active forms, and the former is converted to the latter by the action of a GDP/GTP exchange protein (GEP). We have previously purified a GEP from rat brain with lipid-modified Rab3A as a substrate. Purified Rab3 GEP is active on all the Rab3 subfamily members including Rab3A, -3B, -3C, and -3D. Purified Rab3 GEP is active on the lipid-modified form, but not on the lipid-unmodified form. Purified Rab3 GEP is inactive on Rab3A complexed with Rab GDI. The recombinant protein is prepared from the Rab3 GEP-expressed Spodoptera frugiperda cells (Sf9 cells). The properties of recombinant Rab3 GEP, including the requirement for lipid modifications of Rab3A, the substrate specificity, and the sensitivity to Rab GDI, are similar to those of purified Rab3 GEP. Overexpression of Rab3 GEP inhibits Ca(2+)-dependent exocytosis from PC12 cells. On the other hand, Rab3 GEP is identical to a protein named DENN/MADD: differentially expressed in normal versus neoplastic (DENN)/mitogen-activated protein kinase-activating death domain (MADD). Here, we describe the purification method for recombinant Rab3 GEP from Sf9 cells and the functional properties of Rab3 GEP in Ca(2+)-dependent exocytosis by use of the human growth hormone coexpression assay system of PC12 cells. PMID- 16473593 TI - Biochemical characterization of Alsin, a Rab5 and Rac1 guanine nucleotide exchange factor. AB - Alsin is the gene product mutated in three juvenile-onset neurodegenerative disorders including amyotrophic lateral sclerosis 2 (ALS2). Sequence motif searches within Alsin predict the presence of Vps9, DH, and PH domains, implying that Alsin may function as a guanine nucleotide exchange factor (GEF) for Rab5 and a member of the Rho GTPase family. Procedures are presented in this chapter for the expression, purification, and biochemical characterization of the individual GEF domains of Alsin. A fractionation method is also described for the determination of Alsin's subcellular distribution. The presence of both Rac1 and Rab5 GEF activities makes Alsin a unique dual exchange factor that may couple endocytosis (via Rab5 activation) to cytoskeletal modulation (via Rac1 activation). PMID- 16473594 TI - Purification and analysis of RIN family-novel Rab5 GEFs. AB - The small GTPase Rab5 plays important roles in membrane budding and trafficking in the early endocytic pathways, and the activation of this GTPase is mediated by several guanine nucleotide exchange factors (GEFs) at each of the transport steps. The RIN family has been identified as GEFs for Rab5 and shown to possess unique biochemical properties. The RIN family preferentially interacts with an activated form of Rab5, although it enhances guanine nucleotide exchange reaction. Moreover, biochemical analysis indicates that the RIN family functions as a tetramer. In this chapter, we describe the isolation of the recombinant RIN family via expression in Spodoptera frugiperda (Sf9) insect cells and in mammalian cells. In addition, functional analysis is also provided to assess the physiological properties of the RIN family. PMID- 16473595 TI - Purification and functional properties of a Rab8-specific GEF (Rabin3) in action remodeling and polarized transport. AB - Considering the large number of Rab proteins, only a few Rab-specific exchange factors have been found and characterized. Rab8 is involved in mediating polarized membrane traffic through reorganization of actin and microtubules. It is possible to use the yeast two-hybrid technique to find potential Rab activators. A human protein (Rabin8) and its rat equivalent (Rabin3) were found to bind Rab8 and function as nucleotide exchange factors for Rab8 but not for Rab3A and Rab5. Endogenous and ectopically expressed Rabin8 frequently colocalize with cortical actin. This association is increased by cytochalasin D and phorbol esters that also induced the translocation of both Rabin8 and Rab8 to lamellipodia-like structures. We also show that a GFP-fused Rabin8 behaves identically in this respect. Furthermore, coexpression of Rabin8 with the dominant negative mutant of Rab8 leads to translocation of Rabin8 onto vesicular structures enriched in cell protrusions, indicating that both Rab8 and Rabin8 are involved in mediating polarized membrane transport. This chapter presents a detailed description of the methods and protocols developed to find and characterize a Rab8-specific activator. PMID- 16473596 TI - Assay and functional properties of SopE in the recruitment of Rab5 on Salmonella containing phagosomes. AB - We investigated the mechanism by which Salmonella inhibits transport to lysosomes to survive in macrophages. We have purified the Salmonella-containing phagosomes from macrophages and determined the presence of different endocytic Rab proteins on the phagosomes. Our results have shown that live Salmonella-containing phagosomes recruit more Rab5 than dead Salmonella-containing phagosomes. Recruitment of Rab5 on live Salmonella-containing phagosomes depends on the presence of viable bacteria in the phagosomes. Subsequently, we identified an effector molecule of Salmonella, SopE, which specifically binds Rab5. Moreover, SopE is found to be a specific nucleotide exchange factor for Rab5 and thereby retains Rab5 in an active conformation. Activated Rab5 on Salmonella-containing phagosomes promotes fusion with early endosomes and thus avoids transport to the lysosomes. PMID- 16473597 TI - Purification and functional analyses of ALS2 and its homologue. AB - ALS2 is a causative gene product for a form of the familial motor neuron diseases. Computational genomic analysis identified ALS2CL, which is a novel protein highly homologous to the C-terminal region of ALS2. Both proteins contain the VPS9 domain, which is a hallmark for all known members of the guanine nucleotide exchange factors for Rab5 (Rab5GEF), and are known to act as novel factors modulating the Rab5-mediated endosome dynamics in the cells. It has also been reported that oligomerization of ALS2 is one of the fundamental features of its biochemical and physiological function involving endosome dynamics. This chapter describes methods, including purification of the recombinant ALS2 and ALS2CL, and Rab5GEF assay, which have been utilized to clarify the molecular function for ALS2 and ALS2CL. PMID- 16473598 TI - Polycistronic expression and purification of the ESCRT-II endosomal trafficking complex. AB - Eukaryotic cells use sophisticated mechanisms to direct protein traffic between subcellular compartments. In eukaryotic cells, transmembrane proteins are delivered for degradation in the lysosome or yeast vacuole via multivesicular bodies. The sorting of proteins into lumenal vesicles within multivesicular bodies is directed by the three ESCRT protein complexes. Here we describe the expression and purification of the ESCRT-II complex using the polycistronic expression vector pST39 developed by Tan. In a modification of Tan's procedure, Pfu polymerase amplification with overlapping oligonucleotides was used to generate the translation cassettes for subcloning into pST39 expression vector in a single step. This approach reduces the number of restriction sites and subcloning steps required to express a heterooligomeric protein complex, facilitating rapid screening of multiple complexes and complex variants for crystallization or biochemical characterization. PMID- 16473599 TI - Analysis and properties of the yeast YIP1 family of Ypt-interacting proteins. AB - The YIP1 family of proteins is an intriguing collection of small membrane proteins with critical roles in membrane traffic. Although their mode of action is unknown, they are receiving attention as participants in vesicle biogenesis, and as factors that may mediate the association of Rab proteins with membranes. Yeast YIP1 is an essential gene and can be fully complemented by its human counterpart-suggesting that the essential function of Yip1p is evolutionarily conserved. This chapter presents methods for the cell biological and genetic analysis of Yip1p and other YIP1 family members in the yeast Saccharomyces cerevisiae. PMID- 16473600 TI - Use of Hsp90 inhibitors to disrupt GDI-dependent Rab recycling. AB - Guanine nucleotide dissociation inhibitor (GDI) is a central regulator of Rab GTPase family members. GDI recycles Rab proteins from the membrane and sequesters the inactive GDP-bound form of Rab in the cytosol for use in multiple rounds of transport. The balance between the membrane-bound form of Rab and the cytosolic reserve pool of the Rab-GDI complex is critical for vesicular trafficking between membrane compartments. Recycling of Rab GTPases is likely to require a membrane bound complex of GDI, Hsp90, and Rab given that alphaGDI-dependent recycling of Rab3A at the synapse and neurotransmitter transmitter release is inhibited by Hsp90-specific inhibitors. Here we describe methods required for establishing the dependence of Rab recycling pathways on Hsp90 in vitro. PMID- 16473601 TI - Purification and properties of Yip3/PRA1 as a Rab GDI displacement factor. AB - Prenylated Rab proteins exist in the cytosol bound to guanine dissociation inhibitor (GDI). These dimeric complexes contain all of the information needed for accurate membrane delivery. We have shown that membranes contain a proteinaceous activity that is required for Rab delivery, and we named that activity GDI displacement factor (GDF). Biochemical analysis revealed that GDF activity was membrane associated and had a mass of approximately 25 kDa. We therefore used a candidate gene approach and were able to show that pure Yip3/PRA1 protein displays GDF activity. In this chapter, we review key aspects of GDF analysis: our assay and the method by which we purify Yip3/PRA1 in active form. PMID- 16473602 TI - Purification and analysis of TIP47 function in Rab9-dependent mannose 6-phosphate receptor trafficking. AB - TIP47 (tail interacting protein of 47 kDa) is a cytosolic protein that is essential for the transport of mannose 6-phosphate receptors (MPRs) from endosomes to the trans-Golgi. This protein is recruited from the cytosol onto the surface of late endosomes by Rab9 GTPase, which enables TIP47 to bind to MPR cytoplasmic domains with enhanced affinity. A mutation in a deep hydrophobic cleft of TIP47 (F(236)C) confers enhanced affinity binding to MPR cytoplasmic domains and stabilizes MPRs in living cells. We describe the purification of native and recombinant TIP47 proteins and assays that we use to monitor the function of this protein in MPR transport in living cells. PMID- 16473603 TI - Capture of the small GTPase Rab5 by GDI: regulation by p38 MAP kinase. AB - The small GTPase Rab5 is one of the key regulators of early endocytic traffic and, like other GTPases, cycles between GTP- and GDP-bound states as well as between membrane and cytosol. The latter cycle is controlled by a guanine nucleotide dissociation inhibitor (GDI), which functions as a Rab vehicle in the cytosol. GDI extracts from membranes the inactive GDP-bound form of the Rab. Then, the cytosolic GDI:Rab complex is delivered to the appropriate target membrane, where the Rab protein is reloaded, presumably via a GDI displacement factor (Pfeffer and Aivazian, 2004). We previously reported that the formation of the GDI:Rab5 complex is stimulated by the mitogen-activated protein kinase p38 (Cavalli et al., 2001). Mol. Cell7, 421-432.]. Selective activation of p38 MAPK increases endocytic rates in vivo, presumably allowing more efficient internalization of cell surface components for repair, storage, or degradation. These observations emphasize the possibility that external stimuli contribute to the regulation of membrane traffic. Here, we describe how to monitor the ability of GDI to extract Rab5 from early endosomal membranes in vitro and the role of p38 MAPK in this process. In addition, we detail how to investigate the possible role of p38 MAPK in the regulation of endocytosis in vivo. PMID- 16473604 TI - Rab2 purification and interaction with protein kinase C iota/lambda and glyceraldehyde-3-phosphate dehydrogenase. AB - The small GTPase Rab2 is essential for membrane trafficking in the early secretory pathway. Rab2 associates with vesicular tubular clusters (VTCs) located between the endoplasmic reticulum (ER) and the Golgi complex. VTCs function as transport intermediates and sort anterograde-directed cargo from recycling proteins. Rab2 selectively recruits atypical protein kinase C iota/lambda (aPKCiota/lambda) and glyceraldehyde-3-phosphate (GAPDH) to VTCs where aPKCiota/lambda phosphorylates GAPDH. Both aPKCiota/lambda and GAPDH bind directly to Rab2 and this interaction ultimately results in COPI recruitment and the release of retrograde-directed vesicles. This chapter describes a protocol to purify recombinant Rab2 from Rab2 cDNA transformed bacteria and methods to assess recombinant Rab2 biological activity. Additionally, in vivo and in vitro assays are outlined that are employed to demonstrate Rab2 interaction with the downstream effectors aPKCiota/lambda and GAPDH. PMID- 16473605 TI - Purification and functional interactions of GRASP55 with Rab2. AB - GRASP55 is a member of the GRASP family of peripheral membrane proteins thought to contribute to the organization of the Golgi apparatus. GRASPs are typically found in complex with coiled-coil protein, termed golgins, which interact with specific GTPases of the RAB family. Here I will describe the purification of native GRASP55 complexes from Golgi membranes, and the comparison with the related GRASP65 complex, standard methods for the analysis of interactions between GRASPs, golgins, and Rab GTPases using yeast two-hybrid analysis and protein biochemistry with native and recombinant proteins are also described. PMID- 16473607 TI - Physical and functional interaction of noc2/rab3 in exocytosis. AB - Rab, monomeric small Ras-like GTPase, regulates intracellular membrane trafficking in eukaryotic cells. Rab3 is involved in the exocytotic process in a variety of secretory cells including neuronal, neuroendocrine, endocrine, and exocrine cells. Noc2, originally identified as a molecule homologous to Rabphilin 3, is a putative effector of Rab3. Noc2 interacts with the active (GTP-bound) form of Rab3 and regulates hormone secretion in neuroendocrine and endocrine cells and enzyme release in exocrine cells. This chapter describes two kinds of interaction assay by which the association of Noc2 with Rab3 is analyzed: a yeast two-hybrid assay to detect the interaction of Noc2 with the active form of Rab3 in intact cells and a pull-down assay using GST-fused Noc2 protein to ascertain the physical interaction of Noc2 and Rab3 in vitro. Thus, the Noc2 knockout mouse is a useful model for studying the functional consequences of disruption of the interaction. PMID- 16473606 TI - Purification and properties of rabconnectin-3. AB - Rab3A, a member of the Rab3 small GTP-binding protein (G protein) family, regulates Ca(2+)-dependent exocytosis of neurotransmitter. The cyclical activation and inactivation of Rab3A are essential for the Rab3A action in exocytosis. GDP-Rab3A is activated to GTP-Rab3A by Rab3 GDP/GTP exchange protein (Rab3 GEP), and GTP-Rab3A is inactivated to GDP-Rab3A by Rab3 GTPase-activating protein (Rab3 GAP). We) have found a novel protein, named rabconnectin-3, that is coimmunoprecipitated with Rab3 GEP or GAP from the crude synaptic vesicle fraction of rat brain. Rabconnectin-3 constitutes a subunit structure consisting of alpha and beta subunits and localizes at synaptic vesicles. Overexpression of the C-terminal fragment of rabconnectin-3alpha inhibits Ca(2+)-dependent exocytosis from PC12 cells. We describe the purification method for native rabconnectin-3alpha and -3beta from rat brain and the functional properties of rabconnectin-3alpha in Ca(2+)-dependent exocytosis by use of human growth hormone coexpression assay system of PC12 cells. PMID- 16473608 TI - Functional analysis of slac2-a/melanophilin as a linker protein between Rab27A and myosin Va in melanosome transport. AB - Slac2-a/melanophilin regulates melanosome transport in mammalian skin melanocytes by linking melanosome-bound Rab27A and an actin-based motor protein, myosin Va. Slac2-a consists of an N-terminal Slp homology domain (SHD), which has been identified as a specific GTP-Rab27-binding domain, a myosin Va-binding domain (MBD) in the middle region, and an actin-binding domain (ABD) at the C-terminus. Mutations in the slac2-a/mlph gene cause the abnormal pigmentation (i.e., perinuclear melanosome aggregation in melanocytes) in human Griscelli syndrome type III and in leaden mice because of the inability to form the tripartite protein complex consisting of Rab27A, Slac2-a, and myosin Va. In this chapter we describe the methods, including in vivo melanosome distribution assay combined with dominant-negative approaches and RNA interference technology, that have been used to analyze the function of Slac2-a in melanosome transport in melanocytes. PMID- 16473609 TI - Identification and biochemical analysis of Slac2-c/MyRIP as a Rab27A-, myosin Va/VIIa-, and actin-binding protein. AB - Slac2-c/MyRIP is a specific Rab27A-binding protein that contains an N-terminal synaptotagmin-like protein (Slp) homology domain (SHD, a newly identified GTP Rab27A-binding motif), but in contrast to the Slp family proteins, it lacks C terminal tandem C2 domains. In vitro Slac2-c simultaneously directly interacts with both Rab27A and an actin-based motor protein, myosin Va, via its N-terminal SHD and middle region, respectively, consistent with the fact that the overall structure of Slac2-c is similar to that of Slac2-a/melanophilin, a linker protein between Rab27A and myosin Va in the melanosome transport in melanocytes. Unlike Slac2-a, however, the middle region of Slac2-c interacts with two types of myosins, myosin Va and myosin VIIa. In addition, the most C-terminal part of both Slac2-a and Slac2-c functions as an actin-binding domain: it directly interacts with globular and fibrous actin in vitro, and the actin-binding domain of Slac2-a and Slac2-c colocalizes with actin filaments when it is expressed in living cells (i.e., PC12 cells and mouse melanocytes). In this chapter we describe the methods that have been used to analyze the protein-protein interactions of Slac2-c, specifically with Rab27A, myosin Va/VIIa, and actin. PMID- 16473610 TI - Analysis of the role of Rab27 effector Slp4-a/Granuphilin-a in dense-core vesicle exocytosis. AB - Slp4-a/granuphilin-a is a member of the synaptotagmin-like protein (Slp) family and consists of an N-terminal Slp homology domain (SHD) and C-terminal tandem C2 domains. Slp4-a is specifically localized on secretory granules in some endocrine and exocrine cells through its SHD, and it attenuates Ca(2+)-dependent dense-core vesicle (DCV) exocytosis when transiently expressed in endocrine cells. Although the SHD of Slp4-a interacts with three distinct Rab species (Rab3A, Rab8A, and Rab27A) in vitro, in contrast to other Slp members, which only recognize Rab27 isoforms, Slp4-a functions as a Rab27A effector during DCV exocytosis under physiological conditions. This chapter describes various approaches that have been used to characterize the function of Slp4-a as a Rab27A effector, rather than a Rab3A or Rab8A effector, both in in vitro and in neuroendocrine PC12 cells. Specifically, the methods that have been used to analyze (1) the physical interaction between Slp4-a and Rab27A, including pull-down assay and cotransfection assay in COS-7 cells; (2) the localization of Slp4-a-Rab27A complex on DCVs in PC12 cells; and (3) the involvement of Slp4-a and Rab27A in DCV exocytosis by neuropeptide Y (NPY) cotransfection assay combined with site directed mutagenesis are described. PMID- 16473611 TI - Assay and functional interactions of Rim2 with Rab3. AB - Rim was originally identified as a protein that contains a putative Rab3A effector domain at the N-terminus, the same as rabphilin, and two forms of Rim, Rim1 and Rim2, have been reported in mammals. The putative Rab3A-binding domain (RBD) of Rim consists of two alpha-helical regions (named RBD1 and RBD2) separated by two zinc finger motifs, and several alternative splicing events occur in the RBD1 of both Rims that result in the production of long forms and short forms of RBD. The short forms of Rim2 RBD are capable of interacting with Rab3A with high affinity in vitro, and it is recruited to dense-core vesicles (DCVs) in neuroendocrine PC12 cells through interaction with endogenous Rab3A, whereas the long forms of Rim2 RBD show dramatically reduced Rab3A-binding activity in vitro (more than a 50-fold decrease in affinity compared with the short forms of Rim2 RBD), and it is mainly present in the cytoplasm and nucleus. Expression of the shortest form of Rim2 RBD, but not its Rab3A binding-defective mutant (E36A/R37S), promotes high-KCl-dependent neuropeptide Y secretion from PC12 cells, suggesting that the Rim2 containing the short forms of RBD functions as a Rab3A effector during DCV exocytosis. In this Chapter, I describe several assay methods that have been used to determine the physiological significance of the alternative splicing event in the RBD1 of Rim2, including assays for the in vitro interaction between Rim2 RBD and Rab3A and for the localization of Rim2-RBD on DCVs in PC12 cells. PMID- 16473612 TI - Assay of the Rab-binding specificity of rabphilin and Noc2: target molecules for Rab27. AB - Rabphilin and Noc2 were originally described as Rab3A effector proteins involved in the regulation of secretory vesicle exocytosis in neurons and certain endocrine cells. Both proteins share the conserved N-terminal Rab-binding domain (RBD) that consists of two alpha-helical regions separated by two zinc finger motifs. However, the RBD of rabphilin and Noc2 has been shown to bind Rab27A (the closest homologue of Rab3 isoforms) in preference to Rab3A, both in vitro and in vivo. Rabphilin and Noc2 are recruited to dense-core vesicles (DCVs) in neuroendocrine PC12 cells and regulate their exocytosis through interaction with Rab27A rather than with Rab3A. Rab3A-binding-defective mutants of rabphilin(E50A) and Noc2(E51A) retain the ability to target DCVs in PC12 cells, the same as the wild-type proteins, whereas Rab27A-binding-defective mutants of rabphilin(E50A/I54A) and Noc2(E51A/I55A) do not (i.e., they are present throughout the cytoplasm). Expression of the wild-type or the E50A mutant of rabphilin-RBD, but not the E50A/I54A mutant of rabphilin-RBD, in PC12 cells significantly attenuated DCV exocytosis monitored by high-KCl-stimulated neuropeptide Y secretion. In this chapter we describe various assay methods that have been used to characterize the RBD of rabphilin and Noc2 as "RBD27 (Rab binding domain for Rab27)." PMID- 16473614 TI - Purification and functional properties of Rab11-FIP2. AB - Rab11-FIP2 is a 512-amino acid protein that was first identified in a screen for Rab11 interacting proteins. Database analysis revealed that it belongs to a family of proteins characterized by the presence of a highly homologous domain located at its carboxy-termini. This family was termed the Rab11 family of interacting proteins (Rab11-FIPs), as all members have been demonstrated to interact with Rab11. The Rab11-FIPs can be further subdivided into two classes. Rab11-FIP2 belongs to the class I Rab11-FIPs due to the presence of a C2 domain near its amino-terminus. RCP and Rip11 are the other class I family members. A number of proteins that interact directly with Rab11-FIP2, in addition to Rab11, have been identified. These include the EH domain-containing protein Reps1, the AP-2 subunit alpha-adaptin, the actin-based motor protein myosin Vb, and the chemokine receptors CXCR2 and CXCR4. It is hypothesized that Rab11-FIP2 functions to transport cargo, such as chemokine receptors and the EGF receptor, from the endocytic recycling compartment (ERC) to the plasma membrane. PMID- 16473613 TI - Functional properties of the Rab-binding domain of Rab coupling protein. AB - Rab-Coupling Protein (RCP) is an approximately 80-kDa, hydrophilic protein that belongs to a recently identified family of proteins that is characterized by its ability to interact with Rab11 via a highly homologous Rab-binding domain positioned at its carboxy-termini. Five members of this family have been identified; however, a number of these Rab11-FIPs, including RCP and Rip11, have several splice isoforms. RCP is involved in regulating transport of membrane bound vesicles from the endocytic recycling compartment to the plasma membrane, and it can be found at both locations within the cell. PMID- 16473615 TI - Purification and functional properties of Rab11-FIP3. AB - The Rab family of small GTPases are key regulators of membrane trafficking in eukaryotic cells. Rab11, one member of this family, plays a role in regulating various cellular functions such as plasma membrane recycling, phagocytosis, and cytokinesis. A family of Rab11-binding proteins has been identified and termed the Rab11 family interacting proteins or Rab11-FIPs. Rab11-FIP3, a member of this Rab11-binding protein family, in addition to interacting with Rab11, is also capable of interaction with members of the ADP-Ribosylation Factor (ARF) GTPase family. Here we describe the purification of Rab11-FIP3 and report its biological properties in eukaryotic cells as visualized by immunofluorescence microscopy. PMID- 16473616 TI - Class I FIPs, Rab11-binding proteins that regulate endocytic sorting and recycling. AB - Rab11 GTPase is an important regulator of endocytic membrane traffic. In the GTP bound form Rab GTPases interact with effector proteins and each Rab-effector complex is proposed to regulate a unique trafficking step/event such as vesicle docking, budding, transport, or fusion. At least six Rab11 effectors (family of Rab11 interacting proteins, FIPs) have been identified and shown to interact with Rab11. Based on the sequence homology FIPs are divided in class I and class II subfamilies. Class I FIPs have been hypothesized to regulate the recycling of plasma membrane receptors. In contrast, class II FIPs have been implicated in regulating membrane traffic during more specialized cellular functions, such as cytokinesis. This chapter reviews the background and methodology required for characterizing interactions between FIPs and Rab11, as well as understanding their role in regulating endocytic membrane traffic. PMID- 16473617 TI - Expression and properties of the Rab4, Rabaptin-5alpha, AP-1 complex in endosomal recycling. AB - We previously showed that the small GTPase Rab4 regulates formation of recycling vesicles from early endosomes. To understand how Rab4 accomplishes this task, we started to identify the Rab4 effector protein network. In this chapter, we describe experiments leading to the characterization of a complex consisting of Rab4GTP, its effector Rabaptin-5alpha, and the adaptor protein complex AP-1, which regulates recycling from endosomes. PMID- 16473618 TI - Measurement of the interaction of the p85alpha subunit of phosphatidylinositol 3 kinase with Rab5. AB - During endocytosis of the activated platelet-derived growth factor (PDGF) receptor, phosphatidylinositol 3-kinase (PI3K) remains associated with the receptor. We found that the p85 alpha subunit of PI3 kinase binds directly to Rab5 and possesses GTPase activating protein (GAP) activity toward Rab5. Rab5 is a small monomeric GTPase involved in regulating vesicle fusion events during receptor-mediated endocytosis. We used two methods to characterize the direct binding between Rab5 in various nucleotide-bound states and the p85 protein. In the first assay, the ability of p85 to bind to Rab5 is measured using an enzyme linked immunosorbent assay (ELISA). The second assay is a glutathione S transferase (GST) pull-down approach in which GST-Rab5 proteins in various nucleotide-bound states are allowed to bind p85. In both instances, bound p85 is detected using anti-p85 antibodies. PMID- 16473619 TI - Assay and stimulation of the Rab5 GTPase by the p85 alpha subunit of phosphatidylinositol 3-kinase. AB - Rab5 is a small monomeric GTPase involved in regulating vesicle fusion events during receptor-mediated endocytosis. During endocytosis of the activated platelet-derived growth factor receptor, phosphatidylinositol 3-kinase (PI3K) remains associated with the receptor. We have found that the p85 alpha subunit of PI3K binds directly to Rab5 and possesses GTPase-activating protein (GAP) activity toward Rab5. We describe two methods used to characterize the GAP activity of p85 toward the Rab5 protein. The first method is a steady-state GAP assay, used to show that the p85 alpha protein has GAP activity toward Rab5. The second method is a single turnover GAP assay and measures changes in the catalytic rate of Rab5 GTP hydrolysis with or without the p85 alpha protein. PMID- 16473620 TI - Ubiquitin regulation of the Rab5 family GEF Vps9p. AB - To maintain cellular homeostasis, the levels of transmembrane receptors found on the plasma membrane must be tightly regulated. Endocytosis of activated receptors and the eventual degradation of these transmembrane proteins in the lysosome serve a vital role in maintaining the plasma membrane receptor levels as well as attenuating the downstream signaling pathways. Two processes that regulate this receptor trafficking are the covalent modification of the receptor with ubiquitin (ubiquitylation) and the activation of the Rab5 family of small GTPases. Activation of Rab5 family proteins has been shown to be critical for early steps of the endocytic pathway including delivery of activated receptors to the early endosome, while ubiquitylation of activated receptors has been shown to be involved in receptor internalization, delivery to the endosome, and sorting into the multivesiclar body. In yeast, the guanine nucleotide exchange factor Vps9p serves to integrate the activation of a Rab5 protein (Vps21p) via the Vps9 domain with ubiquitin binding via the CUE domain to facilitate the delivery of ubiquitylated receptors to the endosome. Here we provide detailed protocols for the study of Vps9p in vivo and in vitro with regard to Vps21p activation, ubiquitin binding, and Vps9p ubiquitylation. PMID- 16473621 TI - Analysis of the interaction between GGA1 GAT domain and Rabaptin-5. AB - GGAs are a family of adaptor proteins involved in vesicular transport. As an effector of the small GTPase Arf, GGA interacts using its GAT domain with the GTP bound form of Arf. The GAT domain is also found to interact with ubiquitin and rabaptin-5. Rabaptin-5 is, in turn, an effector of another small GTPase, Rab5, which regulates early endosome fusion. The interaction between GGAs and rabaptin 5 is likely to take place in a pathway between the trans-Golgi network and early endosomes. This chapter describes in vitro biochemical characterization of the interaction between the GGA1 GAT domain and rabaptin-5. Combining with the complex crystal structure, we reveal that the binding mode is helix bundle-to helix bundle in nature. PMID- 16473622 TI - Purification and properties of rab6 interacting proteins. AB - A crucial step in the characterization of novel partners of Rab proteins is the confirmation that they indeed interact together by techniques other than the yeast two-hybrid assay used to discover them. Some methods and clues that would help to discriminate between putative interactors are summarized. Pull-down, co immunoprecipitation, and gel filtration experiments are described as ways of checking protein-protein interaction in vitro and in vivo. PMID- 16473624 TI - Assay and properties of rab6 interaction with dynein-dynactin complexes. AB - RAB GTPases help to maintain the fidelity of membrane trafficking events by recruiting cytosolic tethering and motility factors to vesicle and organelle membranes. In the case of Rab6, it recruits the dynein-dynaction complex to Golgi associated vesicles via an adaptor protein of the Bicaudal-D family. Here we describe methods for the identification of Rab6-binding partners in cell extracts. We then focus on the biochemical analysis of interactions with the dynein-dynactin complex and the adaptor proteins Bicaudal-D1 and -D2. Standard protocols for yeast two-hybrid analysis, and biochemical assays for the analysis of the interactions between Rab6, Bicaudal-D, and the subunits of the dynein dynactin complex are outlined. PMID- 16473623 TI - Affinity purification of Ypt6 effectors and identification of TMF/ARA160 as a Rab6 interactor. AB - Rab/Ypt GTPases are key regulators of intracellular traffic in eukaryotic cells. One important function of Rab/Ypts is the nucleotide-dependent recruitment of downstream effector molecules onto the membrane of organelles. In budding yeast Ypt6 is required for recycling of membrane proteins from endosomes back to the Golgi. A biochemical approach based on the affinity purification of Ypt6:GTP interacting proteins from yeast cytosol led to the identification of two conserved Ypt6 effectors, the tetrameric VFT complex and Sgm1. The mammalian homolog of Sgm1, TMF/ARA160, contains a short conserved coiled-coil motif that is sufficient for the binding to the three mammalian orthologs of Ypt6, Rab6A, Rab6A', and Rab6B. PMID- 16473625 TI - Assay and functional properties of Rabkinesin-6/Rab6-KIFL/MKlp2 in cytokinesis. AB - Here we describe methods for the characterization of the kinesin-9 family motor protein Rabkinesin-6/Rab6-KIFL/MKlp2 in cytokinesis. Here we outline biochemical assays for studying the interaction of Rabkinesin-6/Rab6-KIFL/MKlp2 with microtubules, and its regulation and interaction with the mitotic polo-like kinase 1 using recombinant proteins expressed in and purified from insect cells. Protocols for the in vivo functional analysis of Rabkinesin-6/Rab6-KIFL/MKlp2 using depletion with small interfering RNA duplexes are described. PMID- 16473626 TI - Interaction and functional analyses of human VPS34/p150 phosphatidylinositol 3 kinase complex with Rab7. AB - The Rab7 GTPase is a key regulator of late endocytic membrane transport and autophagy. Rab7 exerts temporal and spatial control over late endocytic membrane transport through interactions with various effector proteins. Among Rab7 effectors, the hVPS34/p150 phosphatidylinositol (PtdIns) 3-kinase complex serves to regulate late endosomal phosphatidylinositol signaling that is important for protein sorting and intraluminal vesicle sequestration. In this chapter, reagents and methods for the characterization of the interactions and regulation of the Rab7/hVPS34/p150 complex are described. Using these methods we demonstrate the requirement for activated Rab7 in the regulation of hVPS34/p150 PtdIns 3-kinase activity on late endosomes in vivo. PMID- 16473627 TI - Functional analyses and interaction of the XAPC7 proteasome subunit with Rab7. AB - Proteasomes have long been known to mediate the degradation of polyubiquitinated proteins in the cytoplasm and the nucleus. Additionally, proteasomes have been identified as participating in cellular degradative pathways involving the endomembrane system. In conjunction with the endoplasmic reticulum, proteasomes serve as a quality control mechanism for disposing of malfolded newly synthesized proteins, while on the endocytic pathway they serve to facilitate the degradation of key signaling and nutrient receptors as well as the destruction of phagocytosed pathogens. Our laboratory has identified a direct interaction between the late endocytic Rab7 GTPase and the alpha-proteasome subunit, XAPC7, thus providing the first molecular link between the endocytic trafficking and cytosolic degradative machineries. In this chapter reagents and methods for studying the regulation and interactions between XAPC7, the 20S proteasome, and Rab7 are described. PMID- 16473628 TI - Expression, assay, and functional properties of RILP. AB - Rab proteins are master regulators of vesicular membrane traffic of endocytic and exocytic pathways. They basically serve to recruit proteins and lipids required for vesicle formation, docking, and fusion. Each Rab protein is able to recruit one or more effectors, and, through the action of effectors, it drives its specific downstream functions. The Rab interacting lysosomal protein (RILP) is a common effector of Rab7 and Rab34, two Rab proteins implicated in the biogenesis of lysosomes. RILP is recruited onto late endosomal/lysosomal membranes by Rab7 GTP where it induces the recruitment of the dynein-dynactin motor complexes. Therefore, through the timed and selective dynein motor recruitment onto late endosomes and lysosomes, Rab7 and RILP control transport to endocytic degradative compartments. A similar role for Rab7 and RILP has been demonstrated also for phagosomes. Indeed, RILP recruits dynein-dynactin motors on Rab7-GTP-positive phagosomes and the recruitment not only displaces phagosomes centripetally, but also promotes the extension of phagosomal tubules toward late endocytic compartments. RILP is therefore a key protein for the biogenesis of lysosomes and phagolysosomes. This chapter describes how to express wild-type or mutated RILP in mammalian cells, and how to test the effects caused by RILP dysfunction. In particular, we report assays to monitor the interaction between RILP and Rab7, morphology and distribution of endosomes, and to measure degradation of endocytic markers. PMID- 16473629 TI - Assay and functional properties of Rab34 interaction with RILP in lysosome morphogenesis. AB - We have recently characterized Rab34 as a new member of the Rab GTPase family based on its ability to regulate lysosomal morphology. Rabbit polyclonal antibody raised against recombinant Rab34 reveals that Rab34 is a 29-kDa protein present both in the cytosol and in the Golgi apparatus. A GTP overlay assay shows that a wild-type and GTP-restricted mutant form of recombinant Rab34 bind GTP in vitro. Yeast two-hybrid interaction screens identify Rab7-interacting lysosomal protein (RILP) as a partner of Rab34. Both GST pull-down experiments and direct binding assays in vitro demonstrate that RILP interacts selectively with the wild-type and GTP-restricted but not GDP-restricted form of Rab34. A key residue (K82) of Rab34 is necessary for interaction with RILP. Expression of EGFP-tagged Rab34 wild-type or GTP-restricted forms in mammalian cells results in redistribution of clustered lysosomes to the peri-Golgi region and this property depends on K82, suggesting that Rab34 regulates lysosome distribution via interaction with RILP. These results suggest that RILP is a common effector shared by Rab7 and Rab34. We describe the methods used in our study. PMID- 16473630 TI - Rabring7: a target protein for rab7 small g protein. AB - Rab7, a member of the Rab family of small G proteins, has been shown to regulate late endocytic traffic and lysosome biogenesis, but the exact roles and the mode of actions of Rab7 are still undetermined. Accumulating evidence suggests that each Rab protein has multiple target proteins and works together with them to coordinate the individual step of vesicle traffic. Rabring7 (Rab7-interacting ring finger protein) is a Rab7 target protein that has been isolated using a CytoTrap system. This protein shows no homology with RILP, which has been reported as another Rab7 target protein. Rabring7 is recruited efficiently to late endosome/lysosome by the GTP-bound form of Rab7. Exogenous expression of Rabring7 not only affects epidermal growth factor degradation but also induces the perinuclear aggregation of lysosomes and the increased acidity in the lysosomes. This chapter describes the procedures for the isolation of Rabring7 with a CytoTrap system, the analysis of the Rab7-Rabring7 interactions, and the properties of Rabring7. PMID- 16473631 TI - Analysis of potential binding of the recombinant Rab9 effector p40 to phosphoinositide-enriched synthetic liposomes. AB - The transport factor p40 is thought to assist Rab9 in mediating late-endosome-to TGN vesicular transport. p40 was recently identified as an associated protein of the PtdIns 5-P/PtdIns 3,5-P2-producing kinase PIKFyve. Moreover, p40 recovery in membrane fractions appeared totally dependent on the presence of an intact enzymatic activity of PIKFyve, implying a mechanism of p40 membrane association dependent on membrane PtdIns 5-P and/or PtdIns 3,5-P2. Here we have evaluated plausible interaction of recombinant p40 with the PIKFyve products PtdIns 5-P and PtdIns 3,5-P2 by a synthetic-liposome binding assay. PMID- 16473632 TI - Assessment of Rab11-FIP2 interacting proteins in vitro. AB - Members of the Rab family of small GTPases are involved in multiple trafficking events in both endocytotic and biosynthetic pathways. To understand more fully the regulation of these events, a concerted effort is underway to ascertain the binding partners and regulators of Rabs. Here, we describe methods to assess binding of Rab11a with Rab11-FIP2 and other Rab11-FIPs utilizing a modified far Western approach. We then broaden this application to assess binding of Rab11 FIP2 with myosin Vb and homodimerization of Rab11-FIP2. PMID- 16473633 TI - Interactions of myosin vb with rab11 family members and cargoes traversing the plasma membrane recycling system. AB - Myosin Vb interacts with Rab11 family members and is the major motor protein identified in association with plasma membrane recycling systems in nonpolarized and polarized cells. Using yeast two-hybrid binary screens, we demonstrated that dominant active Rab25S21V fails to interact with myosin Vb, but does interact with Rab11-FIP2. Transfection of DsRed2-myosin Vb tail in MDCK cell lines stably transfected with wild-type or dominant active forms of Rab11a or Rab25 demonstrated that the distribution of Rab25S21V is only partially altered by expression of the myosin Vb tail. Finally, we demonstrate EGF-dependent sequestration of internalized EGF receptor by EGFP-myosin Vb in A431 cells. Expression of myosin Vb tail represents a powerful provocative test for the trafficking of cargoes through Rab11-containing plasma membrane recycling systems. PMID- 16473634 TI - Properties of Rab13 interaction with protein kinase A. AB - The small GTPase Rab13 localizes to tight junctions in epithelial cells and regulates the recruitment of claudin1 and ZO-1, two proteins required for the assembly of functional tight junctions. Rab13 directly binds to the alpha catalytic subunit of protein kinase A (PKA alpha cat) and reversibly inhibits PKA dependent phosphorylation of vasodilator-stimulated phosphoprotein (VASP), a key actin cytoskeletal remodeling protein. The inhibition of VASP phosphorylation abolishes the targeting of VASP to cell-cell junctions, which in turn leads to a delay in the recruitment of claudin1 and ZO1 into tight junctions. Consequently, tight junctions formed in epithelial cells expressing the GTP-bound Rab13 are structurally disorganized and functionally leaky for small molecules (A. M. Marzesco et al. [2002]. Mol. Biol. Cell13, 1819-1831; K. Kohler et al. [2004]. J. Cell Biol. 165, 175-180). Our data provide the first direct link between activation of small GTPases and the recruitment of cytoskeletal modulators into tight junctions. Here, we describe different procedures we used to demonstrate that Rab13 interacts with PKA and reversibly controls phosphorylation and recruitment of VASP. PMID- 16473635 TI - Functional properties of rab15 effector protein in endocytic recycling. AB - Receptor recycling has emerged as an important regulatory mechanism for cell surface composition, pathogen invasion, and for control over the intensity and duration of receptor signaling in multiple cell types. In the case of the transferrin receptor, receptor recycling is an important step for facilitating iron uptake into the cell, by regulating the availability of the receptor at the cell surface. Following internalization into clathrin-coated pits, the transferrin receptor first enters peripheral sorting endosomes. Here, internalized transferrin receptor is either sorted for recycling back to the cell surface directly, or targeted to a slower route of recycling through a perinuclear population of endosomes termed the endocytic recycling compartment. This chapter describes methodologies to examine the fast and slow modes of transferrin receptor recycling, with a particular emphasis on the function of the novel protein Rab15 effector protein. PMID- 16473636 TI - Assays for interaction between Rab7 and oxysterol binding protein related protein 1L (ORP1L). AB - ORP1L belongs to the recently described family of human oxysterol binding protein homologues. We have previously shown that ORP1L localizes to late endosomes. In this chapter we describe methods that have been used to investigate the functional link of ORP1L with the protein machinery regulating late endosomal membrane trafficking. Co-immunoprecipitation, COS cell two-hybrid, and pull-down assays were applied to demonstrate a physical interaction between ORP1L and the late endosomal small GTPase Rab7. With these methods we were able to map the Rab7 binding determinant of ORP1L to the amino-terminal ankyrin repeat region (aa 1 237) and show that the interaction is preferentially with the GTP-bound form of Rab7. Furthermore, we describe approaches based on transient transfection and confocal immunofluorescence microscopy, which were employed to study the effect of this amino-terminal ORP1L fragment on late endosome morphology. The ankyrin repeat fragment induces juxtanuclear clustering of late endosomes, dependent on an intact microtubule network. When it is coexpressed with the dominant inhibitory Rab7 mutant T22N, the clustering is inhibited, suggesting that the effect involves interaction of the fragment with active Rab7. PMID- 16473638 TI - Purification and functional analysis of a Rab27 effector munc 13-4 using a semi intact platelet dense-granule secretion assay. AB - We have demonstrated that small GTPase Rab27 regulates dense-granule secretion in platelets. Using Rab27a affinity chromatography, we purified Munc 13-4 as a novel Rab27a interacting protein from platelet cytosol. This chapter describes the purification of Munc 13-4 and an in vitro assay system analyzing the mechanism of dense-granule secretion in platelets. The activity of Munc 13-4 is tested in this assay. PMID- 16473637 TI - Characterization of Rab23, a negative regulator of sonic hedgehog signaling. AB - The hedgehog signaling pathway is indispensable in embryogenesis, being responsible for the development of a wide array of vertebrate organs. Given its importance in embryogenesis, the precise regulation of hedgehog signaling is crucial. Aberrant activation of this pathway in postnatal life has been associated with a number of tumor types, reinforcing the role of developmental signaling pathways in tumorigenesis. The small GTPase Rab23 acts as a negative regulator of the hedgehog signaling pathway, most notably in the vertebrate neural system. By analogy with studies of other Rab proteins, analysis of the localization of wild-type and constitutively active and inactive forms of Rab23 provides the potential to shed light on the role of Rab23 at the cellular level. We previously produced expression constructs encoding these proteins for analysis in mammalian cell cultures at both the light and the electron microscopy level. This revealed that both wild-type and active Rab23 localizes to the plasma membrane and to endocytic vesicles (T. M. Evans et al. [2003] Traffic4, 869-884). We describe the methods used to design and make the Rab23 expression constructs, and to assess their localization relative to key hedgehog pathways and endocytic markers in both transiently and stably transfected cell cultures. PMID- 16473639 TI - Analysis of hVps34/hVps15 interactions with Rab5 in vivo and in vitro. AB - The hVps34 phosphatidylinositol (PI) 3-kinase plays an important role in the regulation of vesicular trafficking in the endosomal system. hVps34 associates with a myristylated protein kinase, hVps15. The two proteins are targeting to early endosomal membranes by interactions between hVps15 and activated (GTP bound) Rab5. This leads to the production of the hVps34 product, PI(3)P, in the endosomal membrane, and subsequent recruitment of FYVE and PX domain-containing effector proteins. This chapter describes the analysis of hVps34/hVps15 interactions with Rab5 in tissue culture cells and in vitro. PMID- 16473640 TI - Purification and functional properties of prenylated Rab acceptor 2. AB - PRA2 was found to interact with the ER-localized protein VAMP-associated protein of 33 kDa or VAP-33 by a yeast two-hybrid screen. We describe here the purification of PRA2 and VAP-33 as well as an in vitro pull-down procedure to verify the interaction. PRA2 was found to form a large sodium dodecyl sulfate (SDS)-insoluble complex upon heat denaturation, resulting in significant reduction in the Western immunoblot signal. This phenomenon is specific to PRA2 and was not observed with PRA1. We also found that protein interaction with PRA2 is highly sensitive to detergent and describe a covalent cross-linking procedure for mammalian cell extracts to stabilize the PRA2-containing complex prior to membrane solubilization and immunoprecipitation. PMID- 16473642 TI - Anticancer agents and cardiotoxicity. AB - Although rare, cardiotoxicity is a significant complication of cancer treatment. The incidence and severity of cardiotoxicity are dependent on the type of drugs used, dose and schedule employed, and age of patients, as well as the presence of coexisting cardiac diseases and previous mediastinal irradiation. Anthracyclines are among one of the most active agents in oncology, but their use is often hampered by their cumulative dose-limiting cardiotoxicity. Combination therapy with new drugs in the last decade, such as taxanes and trastuzumab, in the treatment of metastatic breast cancer has yielded impressive results but also unexpected cardiotoxicity. Existing methods of minimizing cardiotoxicity include the use of protective agents such as dexrazoxane, different preparations of anthracyclines such as liposomal formulations, and alternative scheduling techniques. Assessment of left ventricular ejection fraction (LVEF) with two dimensional (2D)-echocardiography or radionuclide ventriculography (RNVG) remains the most pragmatic means of monitoring for cardiotoxicity. The increasing number of long-term survivors of pediatric cancers, as well as the use of trastuzumab, taxanes, and anthracyclines in adjuvant treatment of breast cancer, means that more than ever, cardiotoxicity will remain an important issue for clinicians. PMID- 16473643 TI - Diagnosis, management, and evaluation of chemotherapy-induced peripheral neuropathy. AB - Peripheral neuropathy induced by cancer chemotherapy represents a large unmet need for patients due to the absence of treatment that can prevent or mitigate this common clinical problem. Chemotherapy-induced peripheral neuropathy (CIPN) diagnosis and management is further compounded by the lack of reliable and standardized means to diagnose and monitor patients who are at risk for, or who are symptomatic from, this complication of treatment. The pathogenesis and pathophysiology of CIPN are not fully elucidated, but there is increasing evidence of damage or interference with tubulin function. The diagnosis of CIPN may present a diagnostic dilemma due to the large number of potential toxic etiologies and conditions, which may mimic some of the clinical features; the diagnosis must be approached with care in such patients. The incidence and severity of CIPN is commonly under-reported by physicians as compared with patients. The development of new and reliable methods for the assessment of CIPN as well as safe and effective treatments to prevent this complication of treatment would represent important medical advancements for cancer patients. PMID- 16473644 TI - Hepatotoxicity of chemotherapy. AB - The selection of an antineoplastic regimen for an oncology patient is based first on the availability of effective drugs and then on a balancing of potential treatment-related toxicities with the patient's clinical condition and associated comorbidities. Liver function abnormalities are commonly observed in this patient population and identifying their etiology is often difficult. Immunosuppression, paraneoplastic phenomena, infectious diseases, metastases, and poly-pharmacy may cloud the picture. While criteria for standardizing liver injury have been established, dose modifications often rely on empiric clinical judgment. Therefore, a comprehensive understanding of hepatotoxic manifestations for the most common chemotherapeutic agents is essential. We herein review the hepatotoxicity of commonly used antineoplastic agents and regimens. PMID- 16473645 TI - Renal toxicities of chemotherapy. AB - Chemotherapeutic agents can produce a variety of acute and chronic organ toxicities. Since many antitumor drugs and their metabolites are cleared renally, the kidneys are vulnerable to injury. The drugs involved will determine the site of injury within the kidney, resulting in clinical manifestations ranging from an asymptomatic rise in serum creatinine to acute renal failure. The most common renal toxicities of chemotherapeutic agents are described. PMID- 16473646 TI - Myelotoxicity from chemotherapy. AB - Myelosuppression continues to be a major dose-limiting toxicity for most chemotherapy regimens. While the development of growth factors has changed the approach to myelosuppression, costs remain high and can be measured both in terms of quality of life (QOL) and economic outcomes. Growing data suggest that there may also be effects on response and survival in some tumors. This review will highlight the incidence, effect, and treatment/prevention of myelosuppression and briefly discuss the questions that remain. PMID- 16473647 TI - Dermatologic toxicity of chemotherapeutic agents. AB - Due to its high metabolic rate, skin represents one of the major target organs of chemotherapy-associated toxicity. Reactions range from common, nonspecific exanthematous eruptions to rare but distinctive cutaneous lesions that may not become apparent until a drug transitions from clinical trials to widespread oncologic use. The challenge of the physician is to recognize reaction patterns that reflect a drug reaction, identify a likely causative drug, and determine whether the reaction is a dose-limiting toxicity. This review will focus on the cutaneous side effects of the newer classes of chemotherapy drugs, including targeted monoclonal antibody therapy and small molecule inhibitors. PMID- 16473648 TI - Pulmonary toxicity of chemotherapy. PMID- 16473649 TI - Gastrointestinal toxicity of chemotherapeutic agents. AB - Chemotherapy-induced toxicities commonly occur in sites within the gastrointestinal (GI) tract and account for dose-limiting effects. These toxicities are major contributing factors to dose reduction, delays, and cessation of cancer treatment. Through intensive therapies including surgery, combination chemotherapy, hormonal therapy, and targeted therapy, an increasing number of patients with cancer are experiencing improved survival and long-term disease-free survival, as well as palliation of disease-related symptoms. Thus, GI toxicities should be predicted and appropriate interventions initiated to prevent them when possible and provide effective supportive measures and comprehensive follow-up care. This review will discuss the etiology, incidence, prevention, and treatment of GI toxicities of cancer chemotherapy. PMID- 16473650 TI - Vascular toxicity of antineoplastic agents. AB - Among the various deleterious effects of cancer chemotherapy, vascular toxicity is the least well recognized. This lack of recognition may be because the vasculotoxic phenomena are not unique to antineoplastic agents, can occur in patients without exposure to these agents, and the fact cancer itself may produce a hypercoagulable state. As a result, many vascular events either go unnoticed, are ignored, and/or are attributed to the underlying malignancy. Many antineoplastic therapies are associated with various vascular phenomena that range from simple phelibitis to lethal microangiopathy. Recognition of these events is important to minimize the morbidity and even prevent unnecessary deaths. Herein we review the vascular syndromes that have been reported in association with antineoplastic agents. PMID- 16473651 TI - Extravasation of chemotherapeutic agents: prevention and treatment. AB - Most chemotherapeutic agents can cause varying degrees of local tissue injuries when extravasated. The medical treatment of extravasation is based on proper maintenance of the intravenous (IV) line and application of cold or warm compresses, plus the use of antidotes when available. Antidotes for extravasation that have been shown to be useful are sodium thiosulfate for nitrogen mustard, dimethylsulfoxide for anthracyclines and mitomycin, and hyaluronidase for the vinca alkaloids. New treatments include dexrazoxane, sargramostim, and hyperbaric oxygen for doxorubicin extravasations. Tissue damage secondary to drug infiltration occurs as a result of one of two major mechanisms: (1) the drug is absorbed by local cells in the tissue and binds to critical structures (eg, DNA, microtubules), causing cell death; and (2) the drug does not bind to cellular DNA. Damage to immediately adjacent tissue is more readily neutralized than is damage to surrounding tissue. PMID- 16473652 TI - Evidence for involvement of the phenylpropanoid pathway in the biosynthesis of the norlignan agatharesinol. AB - In order to study the biosynthesis of agatharesinol, a norlignan, l-phenylalanine [ring-2,3,4,5,6-2H] and trans-cinnamic acid-[ring-13C6] were administered to fresh sapwood sticks of Cryptomeria japonica (sugi, Japanese cedar), that is, the labeled precursors were allowed to be absorbed through the tangential section of the wood sticks. The wood sticks were then maintained in high humidity desiccators for approximately 20 d after which ethyl acetate (EtOAc) extracts of the wood sticks were analyzed by gas chromatography-mass spectrometry (GC-MS). Native agatharesinol (trimethylsilylated) produces an m/z 369 ion and an m/z 484 ion that are characteristic of its structure. Agatharesinol formed in the sapwood sticks treated with the deuterium-labeled l-phenylalanine generated both of these ions together with m/z 373 and 377 ions (m/z 369+4 and +8, respectively), and also m/z 488 and 492 ions (m/z 484+4 and +8, respectively). Generation of m/z 373 and 488 ions is attributed to the substitution by deuterium of the four hydrogen atoms of either of the p-hydroxyphenyl rings of agatharesinol, and that of m/z 377 and 492 ions is attributed to the substitution by deuterium of the eight hydrogen atoms of both p-hydroxyphenyl rings. In the administration of the 13C labeled trans-cinnamic acid, m/z 375 and 381 ions (m/z 369+6 and +12, respectively), and also m/z 490 and 496 ions (m/z 484+6 and +12, respectively) were found, indicating that either aromatic ring or both aromatic rings of agatharesinol were 13C-labeled. Consequently, assimilation of the labeled precursors into agatharesinol was clearly detected, and an experimental procedure for studies on the biosynthesis was developed. PMID- 16473653 TI - Pectin methylesterase activity in vivo differs from activity in vitro and enhances polygalacturonase-mediated pectin degradation in tabasco pepper. AB - Polygalacturonase (PG) and pectin methylesterase (PME) activities were analyzed in ripening fruits of two tabasco pepper (Capsicum frutescens) lines that differ in the extent of pectin degradation (depolymerization and dissolution). Ripe 'Easy Pick' fruit is characterized by pectin ultra-degradation and easy fruit detachment from the calyx (deciduous trait), while pectin depolymerization and dissolution in ripe 'Hard Pick' fruit is limited. PG activity in protein extracts increased similarly in both lines during fruit ripening. PME activity in vivo assessed by methanol production, however, was detected only in fruit of the 'Easy Pick' line and was associated with decreased pectin methyl-esterification. In contrast, methanol production in vivo was not detected in fruits of the 'Hard Pick' line and the degree of pectin esterification remained the same throughout ripening. Consequently, a ripening specific PME that is active in vivo appears to enhance PG-mediated pectin ultra-degradation resulting in cell wall dissolution and the deciduous fruit trait. PME activity in vitro, however, was detected in protein extracts from both lines at all ripening stages. This indicates that some PME isozymes are apparently inactive in vivo, particularly in green fruit and throughout ripening in the 'Hard Pick' line, limiting PG-mediated pectin depolymerization which results in moderately difficult fruit separation from the calyx. PMID- 16473654 TI - Shoot-applied polyamines suppress nodule formation in soybean (Glycine max). AB - In legumes, the number of root nodules is controlled by a mechanism called autoregulation. Recently, we found that the foliar brassinosteroid (BR), a plant growth-regulating hormone, systemically regulates the nodule number in soybean plants. In the present study we report that such down-regulation of root nodule formation by a BR may occur through a change of the polyamine contents, with the experimental evidence as follows. The foliar contents of both spermidine (Spd) and spermine (Spm) in the super-nodulating soybean mutant, En6500, were always lower than those in its parent line, Enrei. This lower Spd and Spm content accompanied a striking accumulation of putrescine (Put) in the former plant. This finding indicates that Spd and Spm biosynthesis from their precursor Put is repressed in En6500. The foliar treatments with Spd or Spm of En6500 led to a reduction of both nodule number and root growth. On the other hand, foliar treatment with MDL74038, a specific inhibitor of Spd biosynthesis, apparently increased the root nodule number in Enrei. Foliar application of brassinolide (BL) of En6500 increased the leaf Spd level and reduced the nodule number. These results suggested that BL-induced Spd synthesis in shoots might suppress the root nodule formation. PMID- 16473656 TI - Mycorrhizal symbiosis and response of sorghum plants to combined drought and salinity stresses. AB - Arbuscular mycorrhizal (AM) symbiosis can confer increased host resistance to drought stress, although the effect is unpredictable. Since AM symbiosis also frequently increases host resistance to salinity stress, and since drought and salinity stress are often linked in drying soils, we speculated that the AM influence on plant drought response may be partially the result of AM influence on salinity stress. We tested the hypothesis that AM-induced effects on drought responses would be more pronounced when plants of comparable size are exposed to drought in salinized soils. In two greenhouse experiments, several water relations characteristics were measured in sorghum plants colonized by Glomus intraradices (Gi), Gigaspora margarita (Gm) or a mixture of AM species, during a sustained drought following exposure to salinity treatments (NaCl stress, osmotic stress via concentrated macronutrients, or soil leaching). The presence of excess salt in soils widened the difference in drought responses between AM and nonAM plants in just two instances. Days required for plants to reach stomatal closure were similar for Gi and nonAM plants exposed to drought alone, but with exposure to combined NaCl and drought stress, stomates of Gi plants remained open 17-22% longer than in nonAM plants. Promotion of stomatal conductance by Gm occurred with exposure to NaCl/drought stress but not with drought alone or with soil leaching before drought. In other instances, however, the addition of salt tended to nullify an AM-induced change in drought response. Our findings confirm that AM fungi can alter host response to drought but do not lend much support to the idea that AM-induced salt resistance might help explain why AM plants can be more resilient to drought stress than their nonAM counterparts. PMID- 16473655 TI - Do exogenous polyamines have an impact on the response of a salt-sensitive rice cultivar to NaCl? AB - In order to analyze the putative impact of polyamines (PAs) on the plant response to salt, seedlings from the salt-sensitive rice cultivar I Kong Pao (IKP) were exposed for 5, 12 and 19 days to 0, 50 or 100 mM NaCl in the absence, or in the presence of exogenous PAs (putrescine (Put), spermidine (Spd) or spermine (Spm) 1mM) or inhibitors of PA synthesis (methylglyoxalbis-guanyl hydrazone (MGBG) 1mM, cyclohexylammonium (CHA) 5mM and D-arginine (D-Arg) 5mM). The addition of PAs in nutritive solution reduced plant growth in the absence of NaCl and did not afford protection in the presence of salt. PA-treated plants exhibited a higher K+/Na+ ratio in the shoots, suggesting an improved discrimination among monovalent cations at the root level, especially at the sites of xylem loading. The diamine Put induced a decrease in the shoot water content in the presence of NaCl, while Spd and Spm had no effects on the plant water status. In contrast to Spd, Spm was efficiently translocated to the shoots. Both PAs (Spd and Spm) induced a decrease in cell membrane stability as suggested by a strong increase in malondialdehyde content of PA-treated plants exposed to NaCl. These results are discussed in relation to the putative functions of PAs in stressed plant metabolism. PMID- 16473657 TI - Photosynthetic light and CO2 utilization and C4 traits of two novel super-rice hybrids. AB - Characteristics of photosynthetic light and CO2 use efficiency from seedling to heading stage, and C4 pathway enzyme activities in both flag leaves and lemma were compared between two newly developed super-rice hybrids (Oryza sativa L.), Liangyoupeijiu and Hua-an 3, and a traditional rice hybrid, Shanyou 63. At seedling and tillering stages, Liangyoupeijiu and Hua-an 3 had higher net photosynthetic rates (Pn) and light saturated assimilation rates (Asat) than did Shanyou 63, at both normal (360 micromol mol(-1)) and doubled (720 micromol mol( 1)) CO2 concentrations. At the heading stage, the flag leaves of all three rice hybrids had similar Pn and Asat. However, the two super-rice hybrids had higher apparent quantum yield (AQY) and carboxylation efficiency (CE) during all three typical developmental stages, and higher quantum yield of CO2 fixation (PhiCO2) at the tillering and heading stages. In addition, Liangyoupeijiu showed significantly higher activities of the C(4) pathway enzymes in both flag leaves and lemmas than did Shanyou 63. As a result, flag leaves of the two super-rice hybrids had higher Pn at morning, noontime and late afternoon during the daily cycle. Since most of the grain yield of rice comes from the photosynthesis of flag leaves, the similar Asat and much higher AQY, CE and PhiCO2 at heading stage of the two super-rice hybrids indicates that higher photosynthetic efficiency rather than higher photosynthetic capacity may be the primary factor contributing to their higher grain yields. PMID- 16473658 TI - Induction of beta-glucosidase activity in maize coleoptiles by blue light illumination. AB - The role of beta-glucosidase during the phototropic response in maize (Zea mays) coleoptiles was investigated. Unilateral blue light illumination abruptly up regulated the activity of beta-glucosidase in the illuminated halves, 10 min after the onset of illumination, peaking after 30 min and decreasing thereafter. The level of 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), which is released from DIMBOA glucoside (DIMBOA-Glc) by beta-glucosidase, and its degradation compound 6-methoxy-benzoxazolinone (MBOA) were elevated within 30 min in the illuminated halves as compare to the shaded halves, prior to the phototropic curvature. Furthermore, beta-glucosidase inhibitor treatment significantly decreased the phototropic curvature and decreased growth suppression in the illuminated sides. These results suggest that blue light induces the activity of beta-glucosidase in the illuminated halves of coleoptiles causing an increase in DIMBOA biosynthesis and the growth inhibition that leads to a phototropic curvature. PMID- 16473660 TI - Inhibition of stem elongation in spinach by theobroxide. AB - In the current study, we investigated the influences of theobroxide on stem elongation in spinach (Spinacia oleracea). Our results showed that stem elongation and flower formation were inhibited by spraying spinach plants with theobroxide under inductive, long day conditions (16 h light/8 h dark), while application of exogenous applied GA3 prevented the effect of theobroxide. Quantitative analysis showed that theobroxide suppressed GA1 biosynthesis, whereas the endogenous content of jasmonic acid was unchanged. However, under short day conditions (10 h light/14 h dark), there were no differences in stem length between treated and untreated plants. These results suggest that the inhibition of stem elongation by theobroxide is probably due to the suppression of gibberellin biosynthesis. PMID- 16473659 TI - A novel pathogenesis-related protein (SsPR10) from Solanum surattense with ribonucleolytic and antimicrobial activity is stress- and pathogen-inducible. AB - A cDNA clone (designated as SsPR10, GenBank Accession Number AY660753 ) encoding a PR10 protein from yellow-fruit nightshade (Solanum surattense) was isolated and characterized. SsPR10 encoded a 160-amino-acid polypeptide with a predicted molecular mass of 17.58 kDa and pI of 5.29. Sequence alignments showed that SsPR10 had high identity (68.1%) with CaPR10, but had only about 31.7% identity with JIOsPR10 at the amino acid level. Genomic DNA gel blot analysis indicated that SsPR10 belonged to a multigene family. The constitutively expressed SsPR10 was detected to be the highest in roots of the sterile seedlings cultured in jars, while SsPR10 expression was the highest in old yellow leaves from the seedlings incubated with sap containing TMV. SsPR10 always expressed at slightly higher level in senescent leaves than in tender ones under both conditions. Further expression analysis revealed that the signaling components of defense/stress pathways (MeJA, SA, ABA, GA3, H2O2 and Cu2+) up-regulated significantly the SsPR10 mRNA levels over the control. However, darkness failed to induce SsPR10 expression and its expression was also inhibited by cold treatment. The SsPR10 was successfully expressed in Eschericha coli and the expressed protein was purified to near homogeneity. The dialytically renatured SsPR10 protein without phosphorylation exhibited ribonucleolytic activity against S. surattense leaf total RNA preparations and could inhibit hyphal growth of Pyricularia oryzae. Our findings suggest that the novel stress- and pathogen inducible SsPR10 with ribonucleolytic and antimicrobial activity participates not only in the defense/stress response pathways but also in plants' growth, development and senescence. PMID- 16473661 TI - Cloning of a CACTA transposon-like insertion in intron I of tomato invertase Lin5 gene and identification of transposase-like sequences of Solanaceae species. AB - Very few CACTA transposon-like sequences have been described in Solanaceae species. Sequence information has been restricted to partial transposase (TPase) like fragments, and no target gene of CACTA-like transposon insertion has been described in tomato to date. In this manuscript, we report on a CACTA transposon like insertion in intron I of tomato (Lycopersicon esculentum) invertase gene Lin5 and TPase-like sequences of several Solanaceae species. Consensus primers deduced from the TPase region of the tomato CACTA transposon-like element allowed the amplification of similar sequences from various Solanaceae species of different subfamilies including Solaneae (Solanum tuberosum), Cestreae (Nicotiana tabacum) and Datureae (Datura stramonium). This demonstrates the ubiquitous presence of CACTA-like elements in Solanaceae genomes. The obtained partial sequences are highly conserved, and allow further detection and detailed analysis of CACTA-like transposons throughout Solanaceae species. CACTA-like transposon sequences make possible the evaluation of their use for genome analysis, functional studies of genes and the evolutionary relationships between plant species. PMID- 16473662 TI - Expression analysis of a stress-modulated transcript in drought tolerant and susceptible cultivars of sorghum (Sorghum bicolor). AB - The present study reports the cloning of a 581 bp sequence, designated as SbEST8, from the osmotically stressed germinated seeds of a drought tolerant cultivar of sorghum (Sorghum bicolor). The SbEST8, which shows no homology with the reported gene sequences, is present in multiple copies and lacks restriction fragment length polymorphism among different sorghum cultivars. The expression of SbEST8 in the germinating seeds of sorghum was modulated by different abiotic stresses. Kinetic studies revealed that imposition of osmotic stress after 8h resulted in maximum levels of SbEST8 mRNA in the germinating seeds of cv. ICSV-272, with further stress causing a decline to undetectable levels by 16 h. However, relieving the stress after 12h resulted in an enhancement of SbEST8 mRNA levels for at least another 4h following which it declined. The decrease in SbEST8 mRNA levels in the leaves at 30 DAS in response to drought stress was observed only in the drought susceptible cultivar (CSV-216), whereas its expression was either increased substantially or remained unaffected in the tolerant cultivars, thus suggesting its role in water stress tolerance. PMID- 16473663 TI - Identification of genes induced upon water-deficit stress in a drought-tolerant rice cultivar. AB - Among the abiotic stresses, the availability of water is the most important factor that limits the productive potential of higher plants. The identification of novel genes, determination of their expression patterns, and the understanding of their functions in stress adaptation is essential to improve stress tolerance. Amplified fragment length polymorphism analysis of cDNA was used to identify rice genes differentially expressed in a tolerant rice variety upon water-deficit stress. In total, 103 transcript-derived fragments corresponding to differentially induced genes were identified. The results of the sequence comparison in BLAST database revealed that several differentially expressed TDFs were significantly homologous to stress regulated genes/proteins isolated from rice or other plant species. Most of the transcripts identified here were genes related to metabolism, energy, protein biosynthesis, cell defence, signal transduction, and transport. New genes involved in the response to water-deficit stress in a tolerant rice variety are reported here. PMID- 16473665 TI - Ether a go-go potassium channels and cancer. AB - Ion channels play important roles in health and disease. In the last few years, an interesting relationship between potassium channels and cancer has evolved. Especially, members of the ether a go-go (EAG) potassium channels family have gained interest as research tools for detection and therapy of different cancers. This review will summarize most of the findings relating EAG channels and cancer, focusing on mRNA expression in tissues, oncogenic properties, modulation and pharmacology. Plausible scenarios on the cellular mechanisms of EAG oncogenicity will be discussed. PMID- 16473664 TI - Evidence for a rebinding of antheraxanthin to the light-harvesting complex during the epoxidation reaction of the violaxanthin cycle. AB - In the present study, we investigated the epoxidation reaction of the violaxanthin (Vx) cycle in intact cells of Chlorella vulgaris. Our results show that the overall epoxidation is slightly slower in darkness compared to the epoxidation during high light (HL) illumination. The calculation of the rate constants of the two epoxidation steps revealed that, for both conditions, the first epoxidation step from zeaxanthin (Zx) to antheraxanthin (Ax) is faster than the second epoxidation step from Ax to Vx. However, the most noteworthy result of our present study is that Ax, which is transiently formed during the epoxidation reaction, participates in non-photochemical quenching of chlorophyll fluorescence (NPQ). A correlation between NPQ and the de-epoxidized xanthophyll cycle pigments during the time-course of the epoxidation reaction can only be achieved when NPQ is plotted versus the sum of Zx and Ax. The accumulation of significant amounts of Ax during the epoxidation reaction further indicates that Ax-dependent quenching proceeds with a similar efficiency compared to the Zx-mediated NPQ. As the xanthophyll-dependent NPQ relies on the presence of de-epoxidized xanthophylls in the PS II antenna, Ax-dependent NPQ is only possible under the assumption that Ax rebinds to the light-harvesting complex (LHC) II during the epoxidation reaction. PMID- 16473666 TI - Caveolin-1 as a potential new therapeutic target in multiple myeloma. AB - Caveolae are specialized flask-shaped lipid rafts enriched in cholesterol, sphingolipids, and structural marker proteins termed caveolins. Caveolins are highly conserved hairpin loop-shaped, oligomeric proteins of 22-24 kDa. Besides the plasma cell membrane, caveolins are also present in mitochondria, the endoplasmatic reticulum, the Golgi/trans-Golgi network, and secretory vesicles. They play a critical role in normal vesicular transport, cholesterol homeostasis, and signal transduction. Conversely, dysregulation of caveolin-1 has been associated with several human diseases including multiple myeloma, an incurable malignancy characterized by excess monoclonal plasma cells within the bone marrow. In this mini-review, we characterize the functional role of caveolin-1 in multiple myeloma, and present the preclinical rationale for novel potential therapeutic approaches targeting caveolin-1 in multiple myeloma. PMID- 16473667 TI - A polymorphism in FAS gene promoter associated with increased risk of nasopharyngeal carcinoma and correlated with anti-nuclear autoantibodies induction. AB - Loss of FAS (CD95) expression is a common feature of malignant transformation, which has been related to loss of epithelial cell differentiation and loss of sensitivity to apoptosis. We investigated the potential association between FAS promoter polymorphism and the genetic susceptibility to the Epstein-Barr virus (EBV)-related nasopharyngeal carcinoma. The in vivo functional significance of the FAS polymorphism was investigated by assessing the correlation between FAS genotypes and the presence of autoantibodies to cytoskeleton and nuclear antigens frequently detected in nasopharyngeal carcinoma. We determined the FAS polymorphism distributions by RFLP-PCR in 170 patients with nasopharyngeal carcinoma and in 224 sex and age-matched controls. We used ELISA and the immunofluorescence analysis to characterize the presence of IgG autoantibodies to the cytoskeleton and nuclear proteins in patients' sera. A significantly increased risk of nasopharyngeal carcinoma was associated with heterozygote FAS A/G (OR=2.00, P=0.001) and homozygote FAS-G/G (OR=3.19, P=0.0001) variants. The increased frequency of FAS-G/G genotype is correlated with the presence of anti nuclear autoantibodies in patients with nasopharyngeal carcinoma (P=0.0298). Our results demonstrated that FAS promoter polymorphism was significantly associated with the nasopharyngeal carcinoma in Tunisians. The anti-nuclear autoantibodies induction was also found to be related to FAS polymorphism. The FAS promoter polymorphism associated not only with the increased risk of nasopharyngeal carcinoma in Tunisians but also with immune response deregulation observed in this cancer. PMID- 16473668 TI - FTY720, a fungus metabolite, inhibits invasion ability of androgen-independent prostate cancer cells through inactivation of RhoA-GTPase. AB - The failure of controlling androgen-independent and metastatic prostate cancer growth is the main cause of death in prostate cancer patients. In this study, we have demonstrated evidence on the inhibitory effects of a fungus metabolite, FTY720, on the clonogenesity as well as invasion ability of androgen-independent prostate cancer cells. First, using colony forming assay, we found that FTY720 treatment led to decreased colony forming ability of androgen-independent prostate cancer cell lines DU145 and PC3, indicating its negative role on cancer cell survival. In addition, treatment with relatively low dose of FTY720 (i.e. inhibitory concentration of 50% cell survival) resulted in suppression of prostate cancer cell migration and invasion abilities demonstrated by Wound closure, 3D collagen gel invasion assays and stress fiber staining. Furthermore, we found that the inhibitory effect of FTY720 on prostate cancer invasion was associated with down-regulation of GTP-bound active form of RhoA. Transfection of a dominant-active RhoA vector in DU145 and PC3 cells conferred resistance to FTY720. Since activation of RhoA-GTPase is associated with metastasis in many types of malignancies, our results not only suggest a new agent for the treatment of advanced prostate cancer, but also implicate a possible novel anticancer drug especially against metastatic cancers. PMID- 16473669 TI - Radiobiological effect of 99mTechnetium-MIBI in human peripheral blood lymphocytes: ex vivo study using micronucleus/FISH assay. AB - 99mTc-MIBI is currently used, for cardiac investigations, for parathyroid thyroid imaging and evaluation of various tumours. It has been demonstrated that 99mTc MIBI is specifically taken up by the human peripheral blood lymphocytes (HPBL), cells which are known to be highly radiosensitive. To evaluate the possible chromosomal damage induced on HPBL by their in vitro exposure to increasing activities of 99mTc-MIBI and also to establish whether HPBL undergo apoptosis or necrosis after in vitro exposure to 99mTc-MIBI. Blood from two healthy donors were irradiated, incubated in vitro with increasing activities of 99mTc-MIBI corresponding to absorbed doses ranging from 1 microGy, 100 microGy, 1 cGy, 10 cGy, 50 cGy to 1 Gy. The cytokinesis block micronucleus (MN) assay was used and the frequency of binucleated cells (BN) with MN (MNBN) was analyzed in cultured HPBL (in either the G0- or G1- and S1-phase of the cell cycle). The fluorescence in situ hybridization (FISH) with pancentromeric probes was also applied to study the MN regarding whole chromosomes or acentric fragments. Apoptosis induction by 0.1 Gy of 99mTc-MIBI in HPBL was quantified using annexin-V test. The frequencies of MNBC were similar in control cultures and in HBPL cultures exposed to 1 microGy, 100 microGy and 1 cGy. However, they were significantly higher (P<0.05 versus controls and lower doses) after one treatment exposure to 0.25 mCi of 99mTc-MIBI (corresponding to 10 cGy) or more but the percentages of MNBN with 10 cGy, 50 cGy and 1 Gy did not differ significantly. The increase of MNBN was more pronounced (P<0.05) for cells irradiated during G1 phase than for those irradiated during G0 or S1. Using FISH, 80-90% of the MN were centromere negative. Although small, the absolute number of MN positive for centromeric signal and presumably containing whole chromosomes increased with doses. There is a statistically significant (P=0.001 and 0.006) increase of both apoptotic cells and necrosis, respectively, as compared to control cells in two times studied (24 and 36 h). Chromosomic damages can thus be demonstrated in HPBL after in vitro exposure of blood to at least 0.25 mCi of 99mTc-MIBI corresponding to one absorbed dose of 10 cGy, and for this dose, apoptosis and necrosis phenomenons were detected. PMID- 16473670 TI - Inhibition of cell-cycle progression in HeLa cells by HY52, a novel cyclin dependent kinase inhibitor isolated from Bauhinia forficata. AB - In the course of screening for a novel inhibitor of cyclin-dependent kinase (CDK), HY52 (C17H30O2N2; molecular weight 294) was isolated from the leaves of Pata de Vaca (Bauhinia forficata). The growth of HeLa cells was inhibited in a dose-dependent manner when treated with 0.07 to 0.41 mM of HY52 for 24 h (IC50:0.11 mM). Furthermore, HY52 showed the selective inhibitory activity on CDC2 kinase purified using immunoprecipitation with an IC 50 value of 0.45 mM. A flow cytometric analysis of the HeLa cells treated with HY52 revealed an appreciable cell-cycle arrest in the G1 phase. Moreover, a TUNEL assay exhibited the apoptotic induction of HeLa cells treated with HY52. To obtain further information on the cell-cycle arrest induced by HY52, the expression of certain cell-cycle-associated proteins was examined using a Western blot analysis. The results revealed that HY52 was found to inhibit the proliferation of HeLa cells through inducing a G1-phase arrest by inhibiting pRb phosphorylation via an up regulation of p21WAF1/CIP1 and p27KIP1, and G2/M-phase arrest by down-regulation of CDC2, cyclin A, and cyclin B1. PMID- 16473672 TI - Sonic hedgehog mRNA expression by real-time quantitative PCR in normal and tumor tissues from colorectal cancer patients. AB - Shh is expressed in the early stages of embryogenesis and in the foregut development. Although Shh has been shown to be overexpressed in brain, pancreas, gastric and lung cancers, its role in the development of colorectal cancer has not been examined. We used real-time quantitative PCR to assess Shh mRNA expression levels in tumor and matched normal tissue from 57 colorectal cancer patients and correlated the results with patient clinicopathological characteristics. Shh expression levels were higher in tumor tissue than in normal tissue from the same patient (P=0.00001). Higher levels of Shh expression were associated with early stage disease (P=0.02). Shh overexpression may influence the development of colorectal cancer. PMID- 16473671 TI - Plasma concentration and activity of matrix metalloproteinase 2 and 9 in patients with breast disease, breast cancer and at risk of developing breast cancer. AB - Matrix metalloproteinases (MMPs) are involved in extracellular matrix modification and associated with invasive and metastatic behavior of human malignant tumors. Specifically, MMP2 and MMP9 are implicated in both early and late processes of tumor development. It is reported that MMPs occur as inactive precursors, active enzymes or enzyme inhibitor complexes in biological samples. However, there is limited knowledge on the role of each form in disease and/or the significance of changes in the plasma concentration and/or activity in breast cancer patients. The aim of this study was to determine if patients with breast cancer, benign disease and at risk for developing breast cancer display characteristic levels of active and/or total MMP2 and MMP9 in plasma. Concentration and activity of MMP2 and MMP9 were determined quantitatively in the plasma of 124 female volunteers diagnosed with breast cancer (n=31), benign disease (n=38), or determined by the Gail Model to be at high risk (n=31) or low risk (controls, n=24) of developing breast cancer. Data obtained was statistically analyzed to search for differences/patterns characteristic of each category. Concentration of total MMP2 was significantly lower in control individuals than benign, high risk (P<0.001 respectively) and breast cancer patients (P=0.002). Activity of total MMP2 was significantly lower in controls compared to cancer, benign and high risk patients (P<0.001 respectively). Attempts to build a predictive/descriptive model using canonical discriminant analysis (utilizing all eight features; concentrations and activity levels of active/total MMP2 and MMP9) enabled the distinction of the controls from the high risk, benign and cancer groups. Our results suggest that preoperative plasma concentration and activity of MMP2 and MMP9 may permit sub-classification of female patients with breast disorders. PMID- 16473673 TI - Regulation of hMSH2 and hMLH1 expression in the human colon cancer cell line SW1116 by DNA methyltransferase 1. AB - Aberrant DNA methylation is now recognized as an important epigenetic alteration occurring early in human cancer. To directly study the role of DNA methyltransferase 1 (DNMT1) in the regulation of expression of tumor-related genes in human colon cancer cells, we stably transfected expression constructs containing sense or antisense DNMT1 into the human colon cancer cell line, SW1116. The expression level of mismatch repair genes (MMR), human mut-L homologue 1 (hMLH1) and human Mut S homologue 2 (hMSH2), was monitored by real time RT-PCR. The methylation status of hMLH1 and hMSH2 promoters was determined by bisulfite modification and methylation-specific PCR (MSP). The protein levels of DNMT1, hMSH2 and hMLH1 were determined by Western analysis. The results show that DNMT1 protein expression was increased or decreased in transfected cell lines containing sense or antisense DNMT1 constructs, respectively. In cells expressing the sense DNMT1 construct, the expression of hMLH1 and hMSH2 was down regulated through hypermethylation of their respective promoters. Furthermore, antisense DNMT1 expression induced promoter demethylation and up-regulated transcription of hMSH2 (P<0.05) and hMLH1 (P=0.064) in SW1116 cells. PMID- 16473674 TI - An alternatively spliced transcript of the PHD3 gene retains prolyl hydroxylase activity. AB - Cellular response to limiting oxygen levels is managed, in part, by the transcription factor hypoxia-inducible factor 1 (HIF-1), and the prolyl hydroxylase (PHD) family of oxygen-requiring enzymes. In the process of analyzing the expression of PHD3, we observed the presence of two alternatively processed PHD3 transcripts, designated PHD3Delta1 and PHD3Delta4 . The expression of both PHD3 and PHD3Delta1 was observed in all tissues and cell lines tested, although the expression of the novel PHD3Delta4 appeared to be restricted to primary cancer tissues. The function of PHD3Delta4 was assessed in transfection experiments showing a preserved prolyl hydroxylase activity. We would submit that PHD3 variants generated by alternative splicing may be intrinsically involved in the complex system of oxygen sensing. PMID- 16473675 TI - Otto W. Madelung and the recognition of Madelung's deformity. AB - Madelung's deformity is an uncommon congenital condition of the wrist usually seen in adolescent girls. It first was documented in the 1800s and is characterized by a shortened radius that curves ulnarly and volarly, a prominent ulna head that projects dorsally from the wrist, and a triangular arrangement of the carpal bones. It is mostly an aesthetic deformity although functional problems and pain may prompt surgeons to undertake a variety of corrective surgical procedures with varying degrees of success. This challenging condition is encountered rarely in a hand surgeon's practice. Even more obscure than the condition is the physician it is named after: Otto Wilhelm Madelung, a distinguished and successful German surgeon who lived at the turn of the century. This article provides a historical perspective on the person and the condition that is still an enigma a century later. PMID- 16473676 TI - Combined nerve transfers for C5 and C6 brachial plexus avulsion injury. AB - PURPOSE: To report the results of combined nerve transfer in C5 and C6 brachial plexus avulsion injury. METHODS: Fifteen patients had nerve transfers: spinal accessory nerve to the suprascapular nerve, a part of the ulnar nerve to the biceps motor branch, and the nerve to the long head of the triceps to the anterior branch of the axillary nerve. Patients were evaluated with regard to elbow flexion, shoulder abduction, and shoulder external rotation. RESULTS: All patients had recovered full elbow flexion: 13 scored M4 and 2 scored M3. Thirteen of the 15 patients obtained good results. The weight the patients could lift ranged from 0 to 7 kg. All patients had recovery of the deltoid function: 13 scored M4 and 2 scored M3. All 15 patients achieved useful functional recovery. Ten patients experienced excellent recoveries and 5 were classified as having good results. The mean shoulder abduction was 115 degrees . Shoulder external rotation strength was scored as M4 in 9 patients, M3 in 4 patients, and M2 in 2 patients. The range of motion of external rotation that was measured from full internal rotation averaged 97 degrees . No clinical donor nerve deficits were observed. CONCLUSIONS: We recommend combined nerve transfers for C5 and C6 avulsion root injuries. These nerve transfers have the advantage of a quick recovery time as a result of the short regeneration distance without nerve graft. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic, Level IV. PMID- 16473677 TI - Brachialis muscle transfer to reconstruct finger flexion or wrist extension in brachial plexus palsy. AB - PURPOSE: Tendon transfers are a routine procedure used to improve hand function in brachial plexus injuries; however, muscles from forearm donors are not always available for transfer. In this situation a distant muscle may be used. This study describes transfer of the brachialis muscle to the forearm muscles to reconstruct finger flexion or wrist extension in patients with brachial plexus injuries. METHODS: In 6 patients the brachialis muscle was transferred to the flexor digitorum profundus and the flexor pollicis longus to restore finger and thumb flexion with the goal of reconstructing a key pinch and hook grasp. In 3 patients the brachialis muscle was transferred to the extensor carpi radialis brevis to restore wrist extension. The patients were evaluated at regular intervals and had final assessments between 10 and 12 months after surgery. RESULTS: Brachialis transfer to the flexor digitorum profundus and the flexor pollicis longus resulted in active motion with full range of digital flexion in the 2 patients who had partial flexion before surgery, and for the 4 patients who had no finger flexion before surgery it resulted in a pulp-to-palm distance for the middle finger of 1 cm in 3 patients and of 2 cm in 1 patient. A lateral key pinch and hook grasp reconstruction was achieved in all patients. Grasping and lateral pinch strengths averaged 110 and 94 mm Hg, respectively. When the brachialis was transferred to the wrist extensors the patients recovered 20 degrees of active wrist extension against resistance. CONCLUSIONS: Brachialis muscle transfer to the forearm muscle constitutes a valid strategy in the reconstruction of finger and thumb flexion and wrist extension after brachial plexus injury when forearm donor muscles are not available. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic, Level IV. PMID- 16473678 TI - Pediatric Evaluation of Disability Inventory: its application to children with obstetric brachial plexus palsy. AB - PURPOSE: A standardized method of measurement of self-care ability in children with obstetric brachial plexus palsy (OBPP) has not been universally adopted. A study was conducted to determine if the Pediatric Evaluation of Disability Inventory (PEDI) was able to discriminate between the self-care ability of children with OBPP and their peers and distinguish between those with differing severities of OBPP. METHODS: The PEDI self-care domain results for 45 children with OBPP (30 without hand impairment, 15 with hand impairment) were reviewed retrospectively. RESULTS: The group performance of children without hand impairment was within 1 SD above the mean. The group performance of children with hand impairment was more than 2 SDs below the mean. The difference between the 2 groups was statistically significant. CONCLUSIONS: In this study children without hand impairment did not have a self-care activity limitation as measured by the PEDI. A deficit in self-care ability was found in those with hand impairment. The PEDI was able to differentiate between the performances of reported self-care activities of children with differing severities of OBPP; however, it was unable to discriminate between those without hand impairment and their peers. TYPE OF STUDY/LEVEL OF EVIDENCE: Diagnostic, Level II. PMID- 16473679 TI - Birth brachial plexus injury is the preferred terminology. PMID- 16473680 TI - The heritability of Dupuytren's disease: familial aggregation and its clinical significance. AB - PURPOSE: Dupuytren's disease (DD) is a benign, fibroproliferative disease affecting the hands. The familial occurrence of DD and its presence in identical twins suggests a genetic basis for the condition. Our aims in this study were (1) to provide evidence for familial aggregation of DD by estimating the sibling recurrence-risk ratio and (2) to link previously associated environmental risk factors with family history of DD. METHODS: Patients diagnosed with DD between the ages of 58 and 81 years (N = 92) were interviewed to assess potential risks and the severity of their conditions. A clinical history and examination were performed and we attempted to interview every family member either in person or through a postal questionnaire. RESULTS: The sibling recurrence-risk ratio (lambda(s)) equaled 2.9 and ranged from 2.6 to 3.3 based on the 95% confidence intervals for the population prevalence. This suggests a high genetic basis for the causation of DD. A lower age of onset and greater severity of DD were associated significantly with a positive family history of DD. Other factors showed no statistical significance with familial aggregation of DD. CONCLUSIONS: The familial clustering observed in DD likely is due to genetic influence rather than shared environment, as shown by the lack of association with exposure to environmental risk factors and family history. Understanding the genetic basis of DD is important for developing novel diagnostic, preventative, and therapeutic regimens in the future. TYPE OF STUDY/LEVEL OF EVIDENCE: Prognostic, Level II. PMID- 16473681 TI - Expression of a novel gene, MafB, in Dupuytren's disease. AB - PURPOSE: Dupuytren's disease (DD) is characterized by fibroblastic proliferation of the palmar fascia, often leading to flexion contracture in the hand. Although there is a strong genetic component the genome-wide expression of novel genes is not known. The purpose of this study was to use DNA microarray technology to identify upregulated genes in DD. METHODS: Human tissue samples were harvested from 3 patient sources: DD cord tissue (n = 20), normal-appearing adjacent control fascia (n = 15), and palmar fascia from patients having carpal tunnel release (n = 15). DNA microarray analysis was performed on amplified sample RNA. Novel genes were compared with known gene functions. A candidate gene of interest was studied further by using immunohistochemistry on DD tissue samples and controls. RESULTS: Several novel genes not described previously in the study of DD were upregulated significantly, including MafB, collagen type V, alpha-2 (COL5A2), collagen type VIII, alpha-1 (COL8A1), contactin I (CNTN1), and leucine rich repeat containing 17 (LRRC17). These upregulated genes were compared with their known gene-expression profiles in other tissues and their purported functions. MafB was found to be of particular interest because of its prominent role in tissue development and cellular differentiation. MafB immunohistochemistry showed positive staining in 50% of the DD specimens but complete absence of MafB in all control tissues (adjacent control fascia, carpal tunnel fascia). Co-localization experiments with MafB and alpha-smooth muscle actin showed staining properties in similar regions but these 2 proteins were not confined solely to the same cells. CONCLUSIONS: Microarray analysis of DD tissue has identified significant upregulated gene expression of MafB. MafB protein also is found in Dupuytren's cords but not in control fascia. Co-localization data suggest that the association of MafB with DD is not related exclusively to myofibroblast proliferation. Because of its role in fibroblastic transformation in other models MafB and its relationship to the pathogenesis of DD deserves further study. PMID- 16473682 TI - Revision of thumb trapeziometacarpal arthroplasty. AB - PURPOSE: To determine the incidence of revision surgery after thumb trapeziometacarpal joint arthroplasty and to present the expected results of future surgical treatment. METHODS: We reviewed the patient database at our medical center and found 654 procedures performed between 1988 and 2000 for the treatment of thumb basal joint arthritis. From the database we found that revision arthroplasties were performed on 15 patients with 17 arthroplasties in the treatment of mechanical pain related to instability or bone impingement. Referred patients with primary surgery performed elsewhere and patients with neurogenic pain were reviewed but not included as a primary focus of this study. The revision surgical procedures included soft-tissue interposition alone or soft tissue interposition with ligament reconstruction for mechanical symptoms and neurolysis of peripheral nerves for associated neurogenic pain. The assessment included preoperative and postoperative patient chart review, measurement of pinch and grip strengths, radiographic assessment, and patient contact to provide complete medical information including any further medical or surgical treatment. RESULTS: Based on a grading system previously used to evaluate thumb revision surgery, objective good (or satisfactory) results were found in 13 of 17 revision procedures. The method of soft-tissue revision (resection with soft-tissue interposition or resection with ligament reconstruction) did not influence the outcome of good versus fair or poor results. Persistent failures (2 fair, 2 poor) resulted from both soft-tissue interposition alone and revision with ligament support arthroplasties. Revision arthroplasty with soft-tissue procedures for mechanical pain provided predictably good results unless a nerve injury occurred at the time of the revision surgery. CONCLUSIONS: Failure of primary thumb trapeziometacarpal arthroplasty can be salvaged by a second surgery with ligament reconstruction procedures combined with soft-tissue interposition and provides satisfactory patient outcomes in more than 75% of cases studied. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic, Level IV. PMID- 16473683 TI - Experimental investigation of finger dynamics before and after metacarpophalangeal joint arthroplasty. AB - PURPOSE: The purpose of this study was to quantify the changes in the arc of digital flexion before and after metacarpophalangeal (MCP) silicone arthroplasty with a 30 degrees preflexed design. METHODS: Index, middle, and ring fingers of 4 fresh-frozen cadaver hands were used. Each hand was attached (palmar side up) to a custom test apparatus. The tendon was drawn by a small winch-type servomotor. Micropotentiometers that were attached to the centers of rotation of the MCP, distal interphalangeal, and proximal interphalangeal joints measured angular displacement before and after MCP arthroplasty as a function of tendon excursion. The data were analyzed comparing the angle of flexion initiation and the angular displacement as a function of tendon excursion before and after joint arthroplasty. RESULTS: There were no statistical differences in the angles of the MCP joints at rest, the order of initiation of joint flexion, and the overall degree of flexion between the unoperated fingers and the fingers that had surgery. There was, however, a trend toward delay in flexion initiation, an increase in the MCP angle at rest, and a decrease in torque after implant arthroplasty. CONCLUSIONS: The decrease in initiation of flexion of the MCP joint, although not statistically significant, probably was related to the 30 degrees of preflexion built into the implant. We also noted a trend of decreased flexion at the MCP joint and increased flexion at the proximal interphalangeal and distal interphalangeal joints. This trend may be advantageous in the reconstruction of hands that initially have an MCP joint flexion deformity. TYPE OF STUDY/LEVEL OF EVIDENCE: to come. PMID- 16473684 TI - Reconstruction of thumb loss complicated by skin defects in the thumb-index web space by combined transplantation of free tissues. AB - OBJECTIVE: To introduce a new technique of 1-stage reconstruction for thumb loss complicated by thumb-index web space contracture and to report its clinical effectiveness. METHODS: From November 1994 to September 2004 there were 11 patients with thumb loss and contracture in the thumb-index web space who had a combined transplantation of free tissues to reconstruct the missing thumb and to rebuild the web space. The reconstructive procedure used the second toe and the anterolateral thigh flap in 5 patients, the second toe and the scapular flap in 2, the big toe wraparound flap and the anterolateral thigh flap in 3, and the big toe wrapround flap and the scapular flap in 1. The 2 independent free tissues were connected together by a vascular combination to form an assembly with 1 common vascular pedicle, which then was anastomosed to the selected vessels in the recipient hand. In this vascular series the dorsalis pedis artery and the greater saphenous vein served as the common vascular pedicle of the flap transfers and the radial artery and the cephalic vein provided the recipient vessels. The outcomes of the reconstructions were evaluated using the Michigan Hand Outcomes Questionnaire. RESULTS: The flaps survived completely in all patients except 1. In this patient a small area in the distal part of the transplanted anterolateral thigh flap became necrotic but healed after dressing changes without the need for further surgical intervention. A mean follow-up period of 3.6 years showed a mean increase of 4.3 cm in the width of the thumb index web space and a considerable improvement in overall hand function was noted on the Michigan Hand Outcomes Questionnaire with effect sizes of greater than 3 (large effect) in all domains. CONCLUSIONS: Combined transplantation of the second toe or the big toe wraparound flap and a free skin flap is suitable to reconstruct a missing thumb and repair the associated skin defect in the adjacent thumb-index web space. We found good functional recovery and an acceptable appearance in this series of patients. Type of study/level of evidence: Therapeutic, Level IV. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic, Level IV. PMID- 16473685 TI - Closed treatment of nonrheumatoid extensor tendon dislocations at the metacarpophalangeal joint. AB - PURPOSE: Acute sagittal band injuries at the metacarpophalangeal (MCP) joint resulting in subluxation or dislocation of the extensor tendons may cause pain and swelling at the MCP joint and limit active extension of the MCP joint. These injuries often are treated with surgical repair or reconstruction. This article outlines a nonsurgical treatment protocol that uses a customized splint for acute, nonrheumatoid extensor tendon dislocations caused by injury to the sagittal bands. METHODS: We retrospectively reviewed 10 patients with 11 acute sagittal band injuries who were treated with a splint of thermally molded plastic that differentially holds the injured MCP joint in 25 degrees to 35 degrees of hyperextension relative to the adjacent MCP joints. All the sagittal band ruptures resulted in complete dislocation of the extensor digitorum communis (EDC) tendon-Rayan and Murray type III injuries. Active proximal interphalangeal and distal interphalangeal motion was begun immediately at the time of initial splinting. The average follow-up period was 14 months. RESULTS: At the time of final evaluation all patients had full range of motion in flexion and extension. Eight patients had no pain and 3 had moderate pain. Four patients (5 digits) had no extensor tendon subluxations and 3 had barely discernable subluxations. Three patients had moderate subluxation of the EDC tendon and their treatments were considered failures. One of these patients had subsequent sagittal band reconstruction. CONCLUSIONS: Our results show acute sagittal band injuries in nonrheumatoid patients resulting in dislocation of the EDC tendon can be managed nonsurgically in many patients with a customized splint called the sagittal band bridge. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic, Level IV. PMID- 16473686 TI - Comparison of pullout button versus suture anchor for zone I flexor tendon repair. AB - PURPOSE: To evaluate the clinical outcome after repair of zone I flexor tendon injuries using either the pullout button technique or suture anchors placed in the distal phalanx. METHODS: Between 1998 and 2002 we treated 26 consecutive zone I flexor tendon injuries. Thirteen patients had repairs from 1998 to 2000 using a modified pullout button technique (group A) and 13 patients had repair using suture anchors placed in the distal phalanx (group B). Patient characteristics were similar for both groups. The same postoperative flexor tendon rehabilitation protocol and follow-up schedule were used for both groups. Evaluation included range of motion, sensibility and grip strength, failure, complications, and return to work. The Student t test was used to determine significant differences. RESULTS: All patients completed 1 year of follow-up evaluation. There were 2 infections in group A that resolved with oral antibiotics and no infections in group B. There were no tendon repair failures and no repeat surgeries in either group. At final follow-up evaluation there were no statistically significant differences for the following end points: sensibility (Semmes-Weinstein monofilament testing and 2-point discrimination), active range of motion (at the proximal interphalangeal joint, distal interphalangeal joint, or their combined motion), flexion contracture (at the proximal interphalangeal joint, distal interphalangeal joint, or their combined contracture), and grip strength (injured tendon as a percent of the contralateral uninjured tendon). The suture anchor group had a statistically significant improvement for time to return to work. CONCLUSIONS: There was no significant difference in the clinical outcome after flexor tendon repair using either suture anchors or the pullout button technique. A significant improvement was found for time to return to work for repairs using the suture anchor technique. Flexor tendon repair can be achieved using suture anchors placed in the distal phalanx, thereby avoiding the potential morbidity associated with the pullout button technique. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic, Level III. PMID- 16473688 TI - Anatomic study of arborization patterns of the ulnar artery in Guyon's canal. AB - PURPOSE: To show variations in arborization patterns of the ulnar artery in Guyon's canal and to investigate the relationship between the hypothenar muscles and the ulnar artery. METHODS: Thirty-five embalmed cadaveric hands were dissected and the existence and course of the superficial and deep palmar branches of the ulnar artery and the site of feeding branches to the hypothenar muscles were recorded. The anatomic relationship between the ulnar artery and the hypothenar muscle variations also was investigated. RESULTS: Four arborization patterns were identified. In type 1UA (n = 17 hands), an artery accompanying the deep branch of the ulnar nerve (AADBUN) formed a deep palmar arch (DPA). In type 2UA (n = 11 hands) the AADBUN continued to the feeding artery of the abductor digiti minimi and the distal deep palmar branch of the ulnar artery (DDPBUA) branched off distally. This arterial structure formed a DPA. In type 3UA (n = 6 hands) both the AADBUN and DDPBUA formed DPAs. In type 4UA(n = 1 hand), the AADBUN continued to the feeding artery of the abductor digiti minimi with no DDPBUA and therefore no DPA. A dorsal perforating artery of the ulnar artery also was found in 4 hands. This branch came from the AADBUN at the level of the distal edge of the pisiform and merged with the dorsal carpal arterial arch. We also investigated the relationship between the structural pattern of the hiatus for the deep branch of the ulnar nerve and ulnar artery variation but found no association. The most common pattern observed was a type 1 hiatus with a type 1UA arborization pattern. CONCLUSIONS: Our study confirmed considerable variations in the arborization pattern of the ulnar artery in Guyon's canal. To avoid injury to the arterial branches during surgery in this region care must be taken with respect to variations of the ulnar artery in Guyon's canal. PMID- 16473687 TI - A comparative study of ulnar-shortening osteotomy by the freehand technique versus the Rayhack technique. AB - PURPOSE: To compare the results of ulnar shortening by the traditional freehand method with those achieved by the Rayhack technique. METHODS: A historical cohort of 95 patients (97 ulnas) who had ulnar shortening for the treatment of ulnar impaction syndrome was evaluated. Forty-three patients (45 ulnas) were treated by the freehand technique and 52 patients (52 ulnas) by the Rayhack technique. Both groups were well matched in terms of age, gender, prior history of trauma, and associated injuries. The following variables were compared: duration of surgery, relief of pain, return to work, postoperative complications, time elapsed between surgery and return to work, union of the osteotomy, collinear alignment of the ulnar shaft, and alignment of the plate against the bone. These variables were compared by using the independent-groups t test, chi-square test, and Fisher exact test, as appropriate. RESULTS: Statistical analysis of the compared parameters: duration of surgery, relief of pain, return to work, postoperative complications, time elapsed between surgery and return to work, and union of the osteotomy, showed that none was significant. There were no cases of malalignment of the ulnar shaft or malalignment of the plate against bone in either group. Our calculations show that one would need a cohort of at least 300 patients in each group to show meaningful differences between the groups provided the same proportions held true. CONCLUSIONS: There was a trend toward a higher incidence of nonunion in patients who had the freehand technique although we were unable to show a statistical difference. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic, Level III. PMID- 16473689 TI - Kienbock's disease in a 14-year-old gymnast: a case report. AB - Kienbock's disease is rare in children and there are few reports and therapeutic recommendations in the literature about this condition. We report a case of a 14 year-old female gymnast for whom nonsurgical treatment was followed by complete healing within 12 months. Repeated computed tomography scans provided a sequential coronal, sagittal, and transverse illustration of the progressive healing of the lunate. PMID- 16473690 TI - Reconstruction of the interosseous ligament unloads metallic radial head arthroplasty and the distal ulna in cadavers. AB - PURPOSE: Longitudinal radioulnar dissociation may result when a compressive load to the hand results in excessive proximal migration of the radius with interosseous ligament (IOL) disruption and radial head fracture. Interosseous ligament reconstruction has been proposed to restore more normal forearm mechanics. The objective of this study was to evaluate the effect of IOL reconstruction on compressive load transfer through the forearm after excision and metallic replacement of the radial head in cadavers. METHODS: In 8 cadaveric forearms, 138 N of compressive load was applied to the hand in neutral forearm rotation and neutral elbow varus-valgus alignment. We measured proximal radial migration and 3-dimensional force vectors acting on the distal radius, distal ulna, IOL, proximal radius, and proximal ulna. The experiment was repeated in order for 5 conditions: (1) with the IOL intact, (2) with the IOL reconstructed with a double-bundle flexor carpi radialis construct, (3) with the radial head excised, (4) after metallic radial head arthroplasty, and (5) after cutting the IOL reconstruction. Analysis of variance was used for statistical comparisons. RESULTS: With the IOL intact the resultant load in the distal radius was 94% +/- 3% of hand load, with 75% +/- 2% transmitted to the proximal radius. Double bundle flexor carpi radialis reconstruction effectively restored this relationship. After radial head excision the resultant distal radius load decreased whereas great increases were seen in the resultant distal ulna load, the resultant load in the IOL reconstruction, the resultant load in the proximal ulna, and proximal radial migration. Force continued to transfer through the proximal radius transversely, with a 400% increase in transverse force, a consequence of abutment of the radial stump caused by proximal radioulnar convergence (there was no contact at the capitellum). After radial head arthroplasty loads transferred across the wrist remained closer to intact but the resultant load on the distal ulna and proximal radius remained increased. With cutting of the IOL reconstruction transverse forces in the forearm became negligible, the resultant distal ulna load increased by 50%, and the resultant radial head load increased by 25%. Proximal migration of the radius was small and was increased by 4.6 mm with radial head excision and by 1 mm after radial head replacement compared with the IOL-reconstructed, radial head-intact state. CONCLUSIONS: Interosseous ligament reconstruction may help improve treatment of longitudinal radioulnar dissociation but remains an experimental procedure. PMID- 16473691 TI - Radiofrequency ablation of osteoid osteoma in the upper extremity. AB - PURPOSE: To examine the results of radiofrequency ablation (RFA) of osteoid osteoma in the upper extremity. METHODS: Twenty-nine patients with a clinical and radiographic diagnosis of an upper-extremity osteoid osteoma were referred for treatment between 1990 and 2003. All had computed tomography-guided percutaneous RFA performed by the senior investigator. Records were reviewed for patient age and gender, lesion size and location, prior treatment, and pathology results. Outcomes were determined by questionnaire via mail or telephone. Complete success was defined as complete resolution of pain without further treatment. Partial success was defined as occasional pain with activities that did not require another procedure. Failure was defined as recurrence, no change in symptoms, and/or the need for another procedure. RESULTS: Follow-up data were available for 25 patients at a minimum of 1 year. Lesion locations included 17 humerus, 5 scapula, 2 radius, 1 ulna. Results for 19 patients were rated completely successful, 4 were rated partially successful, and 2 were rated failures. Two patients whose results were not completely successful received decreased RFA temperature or duration because of the proximity of a neurovascular bundle. There were no complications. Outcomes did not correlate with any patient or tumor characteristics. CONCLUSIONS: Radiofrequency ablation is a safe and effective treatment for most osteoid osteomas in the shoulder, arm, and forearm. Lesions that do not allow a safe distance between the electrode and a major neurovascular structure may require surgical excision. This may be of particular importance in lesions of the hand and carpus. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic, Level IV. PMID- 16473692 TI - Use of routine histologic evaluation of carpal ganglions. AB - PURPOSE: Although histologic evaluation is performed routinely on tissue removed during the excision of carpal ganglions, the necessity of this evaluation is uncertain. We evaluated the use of routine histologic evaluation of excised carpal ganglions and performed a cost-benefit analysis. METHODS: A retrospective review identified 160 consecutive carpal ganglion (102 dorsal, 58 volar) excisions performed over the past 7 years by 2 surgeons. The preoperative evaluation, surgical findings, and results of the pathologic assessment were evaluated for patients treated with a typical carpal ganglion excision. RESULTS: In 156 of 160 patients the preoperative and intraoperative data were highly suggestive of a carpal ganglion; the pathology reports confirmed the diagnosis in all of these patients. No new information was provided by pathologic assessment. In 4 patients the preoperative and intraoperative assessments were less certain; the pathologic assessments in these patients also were equivocal. No malignancy or condition requiring additional treatment was identified. At our institution the total cost of pathologic evaluation for a carpal ganglion including processing and interpretation fees is $352. CONCLUSIONS: Routine histologic examination of excised dorsal and volar carpal ganglions may be unnecessary if the preoperative and intraoperative evaluations are consistent with a diagnosis of carpal ganglion. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic, Level IV. PMID- 16473693 TI - Reasons why rheumatoid arthritis patients seek surgical treatment for hand deformities. AB - PURPOSE: Previous studies have found that function and pain are the main factors that persuade physicians to recommend surgical reconstruction to patients with rheumatoid arthritis (RA). The factors that influence patients to choose surgical reconstruction, however, are not known fully. The purpose of this study was to determine how function, pain, and aesthetics rank in order of importance to RA patients who are considering metacarpophalangeal (MCP) joint arthroplasty for rheumatoid hand deformities. METHODS: Study participants are part of a larger National Institutes of Health-sponsored study. Participants who are eligible to receive MCP joint arthroplasty are enrolled in our study using defined inclusion and exclusion criteria. All patients have RA and MCP joint extensor lag and/or ulnar deviation. Study participants choose whether they want to enroll in a surgical group to receive MCP joint arthroplasty or in a nonsurgical group. At enrollment all participants complete the Michigan Hand Outcomes Questionnaire. Function, pain, and aesthetic domains from the Michigan Hand Outcomes Questionnaire were used in a logistic regression model as predictors to determine the factors associated with patients choosing reconstruction for rheumatoid hand deformities. RESULTS: Younger age and female gender were associated significantly with an increased likelihood for choosing MCP joint arthroplasty surgery. The age and gender-adjusted odds ratios of choosing MCP joint arthroplasty were 0.50 for function, 1.47 for pain, and 0.83 for aesthetics. Patients with less function and greater pain were more likely to choose MCP joint arthroplasty. Aesthetic consideration was not a statistically significant predictor. CONCLUSIONS: Impaired function had the strongest association with patients choosing reconstruction and pain relief was the next most important factor. Although aesthetic consideration was less important, it may prove to be an important factor in determining patient satisfaction after surgery. Understanding which factors are associated with choosing rheumatoid hand reconstruction is an essential component of patient preoperative counseling. PMID- 16473694 TI - Investigation of risk acceptance in hand transplantation. AB - PURPOSE: Twenty-four hands have been transplanted in 18 individuals and clinicians continue to debate whether the risks justify the benefits in these procedures. To assess quantitatively the risks versus benefits in hand transplantation we used a validated instrument (Louisville Instrument for Transplantation) to measure the degree of risk individuals are willing to accept to receive different types of transplantation procedures. METHODS: The Louisville Instrument for Transplantation contains 237 standardized questions that incorporate modified standard gamble and time trade-off outcome measures and questions that assess body image perception, depression, self-esteem, optimism, socially desirable responding, and demographics. Respondents were questioned on the extent to which they would trade off specific numbers of life-years or sustain other costs in exchange for receiving 7 different transplantation procedures. For this study we questioned 227 individuals in 4 study populations with differing life experiences: healthy individuals, organ transplant recipients, upper-extremity amputee patients, and lower-extremity amputee patients. RESULTS: All populations questioned perceived risk similarly despite their differing life experiences and would accept differing degrees of risk for the different transplantation procedures. Organ transplant recipients were the most risk-tolerant group whereas upper-extremity amputee patients were the most risk adverse, even when considering a single hand transplant. All groups that were questioned would accept a higher degree of risk to receive a double versus a single hand transplant. CONCLUSIONS: By questioning individuals who have experienced directly the risks of immunosuppression and those who could benefit from hand transplantation, this study presents an empiric basis for assessing risk versus benefit in this procedure. PMID- 16473695 TI - Nerve injuries about the elbow. AB - Multiple neurovascular structures may be at risk during injuries or procedures about the elbow joint. An appreciation of the complex anatomy of the region, the appropriate evaluation procedures and processes to diagnose injury, and an understanding of treatment options are necessary for surgeons who treat elbow injuries. This article reviews the anatomy, diagnosis, and treatment options of injuries to neural structures about the elbow. PMID- 16473696 TI - Radial head arthroplasty. AB - Radial head fractures are a particularly common yet challenging problem facing orthopedic surgeons today. Over the years multiple treatment modalities have been used including conservative management, open reduction and internal fixation, head excision, and radial head arthroplasty. The clinical outcome studies of metallic radial head arthroplasty systems indicate that head replacement is a reasonable option to offer patients with comminuted radial head fractures and complex elbow trauma. Overall the results for this procedure are quite encouraging and appear durable given the severity of the elbow injuries treated with radial head arthroplasty. This article discusses the history, classification, and treatments of radial head fractures. A description of the stabilizing structures of the elbow as they relate to radial head fractures also is included. Surgical considerations that relate to radial head arthroplasty are detailed and encompass arthroplasty indications, surgical technique, postoperative rehabilitation, and complications. Finally the arthroplasty outcomes literature and a review of current implant options also are discussed. PMID- 16473697 TI - Radiology corner: osteoid osteoma of the index finger: case presentation and discussion. PMID- 16473698 TI - Sex and side differences in relative thumb length. PMID- 16473700 TI - Results of four-corner arthrodesis using dorsal circular plate fixation. PMID- 16473703 TI - "Extreme" treatment with an orthodontic component. PMID- 16473705 TI - Implants for anchorage. PMID- 16473706 TI - Effects of different sizes of occlusal metal on curing depth of light-cured orthodontic band cement. AB - INTRODUCTION: Sufficient penetration and quantity of light are necessary to cure band cement. It is doubtful whether this occurs when bands are cemented to teeth restored with large pieces of occlusal metal. The purpose of this in-vitro investigation was to examine the curing depth and degree of conversion from monomer to polymer of light-cured adhesives when used to cement bands to ceramic blocks covered with metal restorations. METHODS: Two resin adhesives--Transbond Plus (3M Unitek, Monrovia, Calf) and Ultra Band-Lok (Reliance Orthodontic Products, Itasca, Ill)--and 3 curing methods were used. The upper surfaces of ceramic blocks (13 x 10 x 15 mm), used instead of human molars, were restored by using metal pieces of different sizes (4 x 8 x 2, 6 x 8 x 2, and 8 x 8 x 2 mm). The curing depth of the cement along the lateral surface of the block was measured at the middle of its width with slide calipers. Fourier transform infrared spectrometry was used to evaluate the degree of conversion of the adhesives. RESULTS: There was unpolymerized resin in the deep area of the cement on curing with central irradiation from the occlusal surface. However, the amount of unpolymerized resin was significantly (P < .05) decreased with the boundary and circle irradiation methods. CONCLUSIONS: The curing depth of band cement was significantly affected by the size of the occlusal metal restoration and the irradiation method. Therefore, a suitable irradiation method is essential for complete curing of resin for cementing bands to teeth with large metal restorations. PMID- 16473707 TI - Bond strength evaluation of ceramic and stainless steel bracket bases subjected to cyclic tensile loading. AB - INTRODUCTION: The aim of this study was to evaluate the effects of loading, static and cyclic, on 1 stainless steel and 3 ceramic bracket bonding bases. METHODS: The brackets were chosen because of the different characteristics of their bonding bases. The brackets in each group were bonded to bovine teeth by using the same adhesive and subjected to both static and cyclic tensile loading; the results were analyzed (ANOVA and Scheffe) for statistical differences among the groups. RESULTS: The analysis showed that ceramic brackets have unique characteristics compared with stainless steel; the most significant is higher bond strengths. The effects of cyclic loading were shown to be significant, in that fatigue testing caused a decrease in mean tensile bond strength for most groups. An additional factor on the bond strength might be the composition and design of the bracket base. CONCLUSIONS: Simulations of occlusal forces (a combination of tensile, shear, and compressive forces) to orthodontic materials during in-vitro studies in accordance with clinical use might adequately describe actual clinical performance. PMID- 16473708 TI - Psychiatry in orthodontics. Part 1: Typical adolescent psychiatric disorders and their relevance to orthodontic practice. AB - Adolescence is a time of rapid physical and mental development. It is also a time when many diagnosable psychiatric diseases are first noticed. A prior study showed that a high rate of suicidal behavior is seen in orthodontic practices. The orthodontist is in a unique position among medical practitioners because treatment occurs over several years with frequent appointments. This article is a current review of the etiology, diagnosis, and therapy for several pertinent mental disorders that occur in adolescents, including mood disorders, schizophrenia, attention-deficit hyperactivity disorder, personality disorders, and eating disorders. All have been associated with high rates of suicidal behavior and completed suicides. With a keen eye for the development of psychiatric issues, the orthodontist is in a position to make appropriate referrals, if needed. PMID- 16473709 TI - Psychiatry in orthodontics. Part 2: Substance abuse among adolescents and its relevance to orthodontic practice. AB - Substance abuse by adolescents is a serious problem that will touch every orthodontic practice. Recent data show that 40% of tenth graders in the United States will use an illicit drug at some time, and 18% will do so in a 30-day period. These are significant figures that should impact orthodontic diagnosis and treatment planning. The nature of orthodontic treatment is unique in that the orthodontist will see relatively healthy adolescent patients on a monthly basis over a period of years. The orthodontist is in a prime position to recognize potential substance abuse problems and make referrals. This article discusses various types of substance abuse, diagnosis, options for referral, and orthodontic implications. PMID- 16473710 TI - Orthodontists and sleep-disordered breathing. PMID- 16473711 TI - Long-term sequellae of oral appliance therapy in obstructive sleep apnea patients: Part 1. Cephalometric analysis. AB - INTRODUCTION: Oral appliances (OAs) have been widely used to treat snoring and sleep apnea, but their effects on craniofacial structures in patients after 5 years or more of wear have not yet been quantified. METHODS: Seventy-one patients who had worn adjustable mandibular repositioners to treat snoring or sleep apnea were evaluated. Upright lateral cephalometric radiographs in centric occlusion taken before treatment and after a mean of 7.3 +/- 2.1 years of OA use were compared. Baseline sleep studies and patient demographic data were included in the analysis. RESULTS: Cephalometric analyses after long term OA use showed significant (P < .01) changes in many variables, including increases in mandibular plane and ANB angles; decreases in overbite and overjet; retroclined maxillary incisors; proclined mandibular incisors; increased lower facial height; and distally tipped maxillary molars with mesially tipped and erupted mandibular molars. The initial deep overbite group had a significantly greater decrease in overbite. Duration of OA use correlated positively with variables such as decreased overbite and increased mandibular plane angle; changes in the dentition appeared to be progressive over time. CONCLUSIONS: After long-term use, OAs appear to cause changes in tooth positions that also might affect mandibular posture. PMID- 16473712 TI - Long-term sequellae of oral appliance therapy in obstructive sleep apnea patients: Part 2. Study-model analysis. AB - INTRODUCTION: Side effects observed in the occlusion and dental arches of patients using an oral appliance (OA) to treat snoring or sleep apnea for more than 5 years have not yet been investigated. METHODS: Stone casts trimmed in centric occlusion before appliance placement and after an average of 7.4 +/- 2.2 years of OA use in 70 patients were compared visually by 5 orthodontists. RESULTS: Of these patients, 14.3% had no occlusal changes, 41.4% had favorable changes, and 44.3% had unfavorable changes. Significant changes in many variables were found. Patients with greater initial overbites and Class II Division 1 and Class II Division 2 malocclusions were more likely to have favorable or no changes. More favorable changes in overbite occurred in subjects with large baseline overbites. A greater baseline overjet and more distal mandibular canine relationship were correlated to favorable changes. A greater initial overjet was correlated to a more favorable change, a decrease in mandibular crowding, a smaller change in anterior crossbite, and a greater change in overjet. CONCLUSIONS: OA wear after a mean of 7.4 years induces clinically relevant changes in the dental arch and the occlusion. PMID- 16473713 TI - Predictors of long-term orthodontic side effects from mandibular advancement devices in patients with snoring and obstructive sleep apnea. AB - INTRODUCTION: Orthodontic side effects can complicate the long-term use of mandibular advancement devices (MADs) in the treatment of patients with snoring and obstructive sleep apnea. The aim of this study was to find predictors of dental side effects from monoblock MADs. METHODS: Four hundred fifty patients, who consecutively received treatment with either soft elastomeric or hard acrylic devices, were followed up after 5.4 +/- 0.8 years (mean +/- SD). The continuing patients responded to questionnaires and had dental examinations and plaster casts made. RESULTS: Twenty-seven patients had moved or died during the follow-up period. Two hundred thirty-six of the remaining 423 patients (56%) continued treatment, and 187 of them reported compliance rates of > or = 50% at night. A small reduction in overjet of < 1 mm was associated with a deepbite with an overbite of > 3 mm and an overjet of < or = 3 mm (odds ratio [OR] = 7.5; P = .015), nasal congestion (OR = 2.9; P = .005), or the use of a soft elastomeric device (OR = 2.7; P = .014) controlled for age, sex, treatment time, and mandibular displacement. A small reduction in overbite of < 1 mm was related to a small opening of the mandible of < 11 mm (OR = 2.5; P = .008). CONCLUSIONS: Orthodontic side effects might be predictable on the basis of initial characteristics in dental occlusion and the design of MADs. PMID- 16473714 TI - A comparison of responders and nonresponders to oral appliance therapy for the treatment of obstructive sleep apnea. AB - INTRODUCTION: This retrospective study compared cephalometric variables between responders and nonresponders to a titratable oral appliance (OA) in a group of subjects matched for sex, pretreatment age, and body mass index (BMI). METHODS: Nine nonresponders as defined by an improvement in the apnea hypopnea index (AHI; <20%) and their individually matched responders were selected for this study. The difference in age for each matched pair was +/-5 years, and, for BMI, the difference was +/-15%. The pretreatment AHI was matched to the same category (moderate, >15 to < or =30; severe I, >30 to < or =45; and severe II, >45 AHI). RESULTS: Middle and inferior airway space and oropharyngeal airway cross sectional area were significantly larger in the nonresponders. Position of the mandible relative to the cervical spine was the only significant skeletal variable and was larger in nonresponders. Changes in BMI between the groups were statistically significant; the averages were a 2.9% increase in the nonresponders and a 0.5% decrease in responders. The wider airway in nonresponders might reflect an enhanced neuromuscular compensation while awake. The weight gain in nonresponders was relatively small, but it might have reduced the effectiveness of the OA. CONCLUSION: When treating OSA patients with OA therapy, clinicians should pay particular attention to airway size and weight changes. PMID- 16473715 TI - Factors influencing treatment time in orthodontic patients. AB - INTRODUCTION: The purpose of this retrospective study was to identify (and quantify the effect of) factors that influence orthodontic treatment time. METHOD: The sample consisted of 366 consecutive orthodontic patients (220 female, 146 male, aged 10-20 years, treated by 1 orthodontist) who completed treatment in a single stage with fixed appliances. Four categories of data (sociodemographic characteristics, malocclusion characteristics, treatment methods, and patient cooperation) were collected from treatment records and analyzed. RESULTS: Average treatment time was 23.5 months (range, 12-37; SD, 4.7). A multiple regression model explained 38% of the variance and identified 9 significant variables. Five were pretreatment characteristics: male sex, maxillary crowding of 3 mm or more, Class II molar relationships, proposed treatment plan involving extractions, and delayed extractions. The remaining variables (3 of which were associated with patient cooperation) were poor oral hygiene, poor elastic wear, bracket breakages, and brackets rebonded for repositioning. CONCLUSIONS: Orthodontic treatment time is influenced by a number of patient characteristics and clinical decisions. It is possible to predict estimated treatment time for a patient by using a small number of personal characteristics and treatment decisions. PMID- 16473716 TI - Clinical and quantitative assessment of headgear compliance: a pilot study. AB - INTRODUCTION: This study was undertaken to evaluate the compliance of patients using headgear with a timing device and to determine the efficiency of the electronic module timer as a patient motivator. METHODS: Twenty-one patients (average age, 14 years 10 months) were selected from the orthodontic clinic of Federal University of Parana on the basis of headgear wear for anchorage. The patients were instructed to wear their headgear 14 hours a day for a given number of days. The headgears were equipped with recorders (Compliance Science System and Affirm smart headgear modules, Ortho Kinetics, Vista, Calif). The patients were not told about the recorders, but they were instructed to keep track of their wear times. At the end of the test period (T1), the patients' reported wear times were compared with readings from the electronic modules. The patients were assigned a second period of headgear wear (T2) and told that their use would be monitored electronically. Again, the wear times reported by the patients were compared with the values from the electronic modules. Total time, number of hours accumulated between sessions, and quality time (uninterrupted use of headgear) were assessed. RESULTS: Patients reported wearing their headgear an average of 13.6 hours per day; the electronic modules detected 5.6 hours per day in T1 and 6.7 hours per day in T2. Quality time was 1.8 hours per day in T1 and T2. The mean actual hours of daily wear relative to the provider's requirement was 56.7% in T1 and 62.7% in T2. Boys were more compliant than girls. After they learned of the electronic device, the girls' compliance improved. Younger patients were more compliant than older ones. The compliance rate of older patients improved slightly in T2. CONCLUSIONS: Patients tend to overreport their headgear wear times. The mean actual hours of daily wear relative to the providers' requirement was 56.7%. This increased to 62.7% when patients knew a recording device was being used. A monitoring system can provide feedback to the patient, facilitate parental involvement, and motivate patients to comply with headgear wear. PMID- 16473717 TI - Antimicrobial properties of an orthodontic adhesive combined with cetylpyridinium chloride. AB - INTRODUCTION: Patients undergoing fixed appliance treatment are at greater risk for increases in salivary and plaque levels of Streptococcus mutans and an elevated risk of dental caries. Cetylpyridinium chloride (CPC) is known to be an effective antiplaque agent. The purpose of this study was to investigate whether incorporating CPC into a commercially available orthodontic adhesive would impart antimicrobial properties without altering the diametral tensile strength of the material. METHODS: CPC was added to a commercially available, filled, photo activated bracket adhesive (Transbond XT, Unitek 3M, Monrovia, Calif) in varying amounts, to obtain specimens with CPC concentrations of 0% (control), 2.5%, 5.0%, and 10.0% by weight. Adhesive discs 2 mm thick and 4 mm in diameter were incubated with Streptococcus mutans for 48 hours. The diameters of the zones of bacterial inhibition were measured in an agar disc diffusion assay; specimens of each concentration were tested every other week for 196 days. Other discs were soaked in distilled water for 180 days. The amount of CPC released into the water from the modified discs was measured and recorded on days 7, 15, 30, 60, and 180 by using a spectrophotometer at 254 nm. Diametral tensile strength of the modified adhesive discs was measured with a universal testing machine, and the effect of water aging was also evaluated. RESULTS: The measured zone of bacterial inhibition increased as CPC content increased. All CPC-adhesive specimens maintained antimicrobial activity up to 196 days. No zone of bacterial inhibition was measured around the control specimens. CPC release was observed through the end of 180-day period, but the greatest release was recorded in the first week. There was no significant difference (P < .05) in diametral tensile strength between the 2.5% CPC-adhesive group and the control; there were significant differences among the 5.0% and 10.0% CPC-adhesive groups and the control. Water aging had no significant effect on diametral tensile strength other than decreasing it for the test group containing 10.0% CPC. CONCLUSIONS: The incorporation of 2.5% CPC in adhesive material imparted antimicrobial activity without altering diametral tensile strength. PMID- 16473718 TI - Inhibition of orthodontically induced root resorption with echistatin, an RGD containing peptide. AB - INTRODUCTION: Induced dental root resorption is a common side effect of orthodontic treatment. It is an unpredictable phenomenon, and its etiology is unknown. Odontoclasts responsible for the resorption of the dental tissues--ie, cementum and dentin--share many cytochemical and morphological characteristics with osteoclasts, which are responsible for bone resorption. The aim of this study was to explore cellular mechanisms that decrease induced root resorption in orthodontically treated teeth. METHODS: The effects of targeting the alphavbeta3 integrin receptor, expressed by odontoclasts, on induced root resorption surface areas and the number of root resorption lacunae were investigated by using an RGD containing peptide, echistatin. The effect of echistatin on the number of clast cells in the periodontium was also examined. Tooth movement was achieved in 14 Sprague-Dawley rats by placing elastic bands between the right maxillary first and second molars for 24 hours. The animals were equally divided into 2 groups; the experimental animals received echistatin intravenously for 8 hours (0.8 microg/kg/min), and the controls received sterile water. The specimens obtained were processed for light microscopy. The surface area and the number of root resorption lacunae were measured histomorphometrically by using digital photomicrographs. Echistatin labeled with a fluorescent marker was used to confirm its presence in clast cells with fluorescent microscopy. Cytochemically, tartrate-resistant acid phosphatase was used to quantify mature and committed clast cells. Echistatin was localized in targeted cells in the periodontium. RESULTS: Echistatin significantly decreased root resorption surface areas (P < .01) and reduced the number of root resorption lacunae (P < .01). There was no statistically significant difference in clast cell numbers. CONCLUSIONS: Targeting alphavbeta3 integrin receptor expressed by odontoclasts can be effective in reducing root resorption during tooth movement. Further studies are needed to elucidate the mechanism of this inhibition. PMID- 16473719 TI - Effect of debonding force direction on orthodontic shear bond strength. AB - INTRODUCTION: The aim of this study was to determine the influence of debonding force direction in shear bond strength testing of orthodontic brackets in vitro. METHODS: One hundred fifty extracted bovine permanent mandibular incisors were randomly divided into 5 groups of 30 specimens each. Before bonding with a composite adhesive, enamel surfaces were etched with 37% phosphoric acid for 30 seconds. Teeth were bonded with mesh-based stainless steel orthodontic brackets. Shear bond strength was measured with a universal testing machine. Debonding forces were directed either parallel to the bracket base (group B), toward the enamel surface (group A: 15 degrees), or away from the enamel surface (group C: 15 degrees; group D: 30 degrees; group E: 45 degrees). RESULTS: Shear bond strength measurements were as follows: 22.90 (SD, 1.72) MPa (group A), 17.90 (SD, 2.63) MPa (group B), 12.99 (SD, 1.71) MPa (group C), 7.93 (SD, 1.14) MPa (group D), and 6.65 (SD, 1.14) MPa (group E). Analysis of variance indicated that shear bond strengths were significantly different between the groups investigated. Mean adhesive remnant index scores ranged between 1.60 in group B and 2.13 in group D. CONCLUSIONS: Shear bond strength measurements were significantly influenced by the direction of the debonding force, indicating the need for control and standardization of this testing parameter in orthodontic shear bond strength testing. PMID- 16473720 TI - Effect of an antioxidizing agent on the shear bond strength of brackets bonded to bleached human enamel. AB - INTRODUCTION: The purpose of this study was to investigate the effect of antioxidant treatment and delayed bonding on the shear bond strength of metal brackets bonded with composite resin to human enamel after bleaching with carbamide peroxide (CP). METHODS: Eighty recently extracted premolars were divided into an experimental group (n = 60), which was bleached with 10% CP, and a control group (n = 20), which was not bleached. The experimental group was further divided into 3 groups. Specimens in group 1 (n = 20) were bonded immediately after bleaching; specimens in group 2 (n = 20) were bleached, then treated with 10% sodium ascorbate, an antioxidant agent, and then bonded; group 3 specimens (n = 20) were bleached, then immersed in artificial saliva and held for 1 week before bonding. The specimens were debonded, and the enamel surfaces and bracket bases were examined with a stereomicroscope. The adhesive remnant index was used to assess the amount of resin left on the enamel surfaces after debonding. The shear bond strength data were subjected to 1-way analysis of variance. Multiple comparisons were performed with the Bonferroni test. The level of significance was established at P < .05 for all statistical tests. RESULTS: Shear bond strength of brackets bonded immediately after bleaching with 10% CP was significantly lower than that of brackets bonded to unbleached enamel (P < .05). No statistically significant differences in shear bond strength were noted when the antioxidant-treated and delayed bonding groups were compared with the control group (P > .05). CONCLUSIONS: Bleaching with 10% CP immediately before bonding reduces the bond strength of composite resin to enamel. Treating the bleached enamel surface with 10% sodium ascorbate or waiting 1 week reverses the reduction. PMID- 16473721 TI - Effect of chlorhexidine application on shear bond strength of brackets bonded with a resin-modified glass ionomer. AB - INTRODUCTION: The purpose of study was to assess the effect of chlorhexidine application on the shear bond strength and bond failure site of a resin-modified glass ionomer (Fuji Ortho LC, GC Europe, Leuven, Belgium). METHODS: Forty-five bovine permanent mandibular incisors were randomly divided into 3 groups of 15. Group 1 had no chlorhexidine treatment and served as the control; groups 2 and 3 had chlorhexidine application immediately before and 1 week before bonding, respectively. Stainless steel brackets (DB, Leone, Sesto Fiorentino, Italy) were bonded with the resin-modified glass ionomer. After bonding, all samples were stored in distilled water for 24 hours and subsequently tested in a shear mode on a testing machine. Analysis of variance (ANOVA) and the Scheffe test were applied to determine significant differences in the shear bond strength values, and the chi-square test was used to determine significant differences in the adhesive remnant index scores among the groups. Significance for all statistical tests was predetermined at P < .05. RESULTS: Group 1 (no chlorhexidine application) showed significantly higher shear bond strength values than group 2 (chlorhexidine applied immediately before bonding). No significant differences were found between groups 1 and 3 (chlorhexidine applied 1 week before bonding). Moreover, significant differences in debond locations were found among the 3 groups. CONCLUSIONS: Chlorhexidine application immediately before bonding significantly lowered the bond strength values of Fuji Ortho LC but did not affect its bond strength when applied 1 week before bonding. PMID- 16473722 TI - Comparison of bond strength between orthodontic brackets bonded with halogen and plasma arc curing lights: an in-vitro and in-vivo study. AB - INTRODUCTION: This study assessed in-vitro shear bond strength and in-vivo survival rate of orthodontic brackets bonded with either a halogen or a plasma arc light. METHODS: Ninety extracted premolars were divided into 6 groups of 15. Stainless steel brackets were bonded to the teeth by using either a halogen light with a 20-second curing time or a plasma arc light with a 2-, 6-, or 10-second curing time. Brackets were debonded either within 30 minutes of bonding or after thermocycling for 24 hours. Bond strengths were tested on a testing machine at a crosshead speed of 1 mm/minute. The bracket failure interface was measured with a modified adhesive remnant index score. Data were analyzed by using ANOVA and Tukey-Kramer multiple comparison tests. For the in-vivo study, a split-arch design was used to determine the bracket-failure rate and distribution in 25 patients. The patients were followed for a mean period of 1.1 years (386 days). Survival analysis was carried out to compare the failure rates of the 2 techniques. RESULTS: No significant differences in bond strengths were found 30 minutes after bonding between the halogen light (13.6 +/- 3.8 MPa) and the plasma arc light with 2-, 6-, or 10-second curing times (9.6 +/- 2.9, 14.2 +/- 4.6, 16.0 +/- 3.0 MPa, respectively). Similar bond strengths were also found between the halogen light with a 20-second (16.1 +/- 3.6 MPa) curing time and plasma arc light with 6 seconds (18.2 +/- 4.6 MPa) of curing time after 24 hours of thermocycling. For the in-vivo study, no significant difference was found in bracket failure rates between the 2 light sources (4.9% in both groups). No significant differences were found between ARI scores for the halogen light and the plasma arc light at either 30 minutes or 24 hours after debonding. CONCLUSIONS: These results indicate that the plasma arc light with a 6-second curing time can produce similar bond strength and bracket-failure rates as the halogen light that requires a longer curing time. PMID- 16473723 TI - Correction of severe obstructive sleep apnea with bimaxillary transverse distraction osteogenesis and maxillomandibular advancement. AB - Obstructive sleep apnea (OSA) can be a debilitating, even life-threatening, condition. The most favorable treatment for patients with OSA is multidisciplinary care by a team that represents various dental and medical disciplines. Prescribed therapies might include weight loss, behavior modification, oral appliances, soft tissue surgery, skeletal surgery, or some combination of approaches. When orthognathic surgery has been used, often only the anteroposterior dimension is addressed, and the transverse dimension is overlooked. The treatment presented here demonstrates the important role that transverse expansion of the maxillary and mandibular arches can have for patients with severe OSA. An initial stage of maxillary and mandibular transverse distraction osteogenesis was performed, followed immediately by fixed orthodontic treatment. After appropriate orthodontic alignment, leveling, and coordination, a second surgical stage consisting of maxillary and mandibular advancement was performed. Marked enhancements in occlusion and facial morphology, and a profound improvement in the OSA, were obtained. PMID- 16473724 TI - Orthodontics in 3 millennia. Chapter 7: Facial analysis before the advent of the cephalometer. AB - The cephalometer was not invented in a vacuum. It was the culmination of centuries of efforts on the part of artists, anthropologists, and scientists to fathom nature's vicissitudes. Whereas Renaissance investigators "caged" the human face in a series of grids in an effort to find proportional relations, 20th century orthodontists were more interested in knowing how the teeth and jaws related to the face and cranial base. Primarily a research tool, the cephalometer became a means of unmasking a patient's whole developmental pattern, becoming our most important diagnostic tool since study models. PMID- 16473725 TI - Articulators in orthodontics: an evidence-based perspective. PMID- 16473726 TI - Color management for perfect color images. PMID- 16473728 TI - Reliability and validity of pinch and thumb strength measurements in de Quervain's disease. AB - The purpose of this study is to evaluate the test-retest reliability and construct validity of pinch and thumb strength measurements in subjects with de Quervain's disease. Maximal palmar pinch and thumb strength (adduction, extension, abduction, and flexion) were measured using, respectively, a pinch gauge and a biaxial dynamometer. The reliability was estimated using the generalizability theory. The validity hypotheses were as follows: 1) the pinch and thumb strength of the symptomatic side would be significantly lower than that of the asymptomatic side, and 2) the strength loss would be greater for thumb extension and abduction. The reliability was high for all strength measurements, pinch strength being the more reliable one. The pinch and thumb strength in all directions evaluated was significantly decreased on the symptomatic side (p<0.003); no direction showed a greater decrease than the others. The results suggest that pinch and thumb strength measurements are reliable and able to show a decreased strength on the symptomatic side in this population. PMID- 16473729 TI - Reliability and validity of a self-report of hand function in persons with rheumatoid arthritis. AB - The purpose of this study was to examine the test-retest reliability and the concurrent validity of the Duruoz Hand Index (DHI) in persons with rheumatoid arthritis (RA). Forty participants with RA and no other major medical problems completed the DHI, a self-report of hand function, at two points in time to assess test-retest reliability. To determine concurrent validity, participants were also administered three performance-based tests, the Arthritis Hand Function Test (AHFT), the Hand Mobility in Scleroderma Test (HAMIS), and the Keitel Functional Test (KFT), and two self-report questionnaires of functional ability, the Health Assessment Questionnaire (HAQ) and the Scleroderma Functional Assessment Questionnaire (SFAQ). Test-retest reliability intraclass correlation coefficients for the DHI ranged from 0.83 to 0.90. Scores on the DHI were significantly correlated with scores on the AHFT (r(s)=0.36-0.54), the HAMIS (r(s)=0.39), the HAQ (r(s)=0.78), the HAMIS (r(s)=0.39), and the SFAQ (r(s)=0.85). Scores on the DHI did not correlate with KFT scores. The results from this study show the DHI to be a reliable and valid test for hand function in persons with RA. PMID- 16473730 TI - Construct validity of the Chinese version of the Patient-rated Wrist Evaluation Questionnaire (PRWE-Hong Kong Version). AB - With increasing economic globalization, including health care, it is important to use standardized outcome measures applicable to a broad spectrum of patients in a wide array of countries. The purpose of this study was to verify construct and content validity and reliability of the Chinese version of the Patient-rated Wrist Evaluation Questionnaire (PRWE-Hong Kong version). The PRWE was translated into Chinese, and face validity was established by inviting experts and patients to participate in the panel review of the questionnaire. A correlation field study was performed using a convenience sample of 47 patients with wrist injuries. Patients were assessed at baseline and six weeks after the initial measurement. The following measures were taken: the Chinese version of the PRWE and the Medical Outcome Short Form (36) Health Survey (SF-36), Visual Analogue Scale (VAS) for pain, active wrist range of motion, grip strength, and the Jebsen Hand Function Test. Statistical analysis consisted of Pearson correlation coefficients (convergent validity), factor analysis (content validity), paired t test (convergent validity), and the Cronbach alpha (internal consistency). Clinically relevant correlations existed between "Pain at rest" and the VAS "resting pain" (r=0.785, p<0.0001) as well as between "Pain on repeated wrist movement" and the VAS "exertion pain" (r=0.872, p<0.0001). The "Physical Component Summary" of the SF-36 was found significantly correlated with the PRWE function subset total score (r=-0.618, p<0.0001), and the PRWE total score (r= 0.645, p<0.0001). The specific function subset score also correlated with the wrist flexion range (r=-0.308, p<0.0001) and the grip strength (r=-0.488, p=0.035). Two factors were found that accounted for 61% of the variance. The Cronbach alpha coefficients ranged from 0.7805 to 0.9502, indicating that the internal consistency of the questionnaire items was sound and reliable. Positive correlations between the wrist ranges of motion (ROM) and the specific function subset score showed that the function subset measured dimensions related to wrist specific performance in activities of daily living. Factor analysis results supported the construct validity of the PRWE-Hong Kong version in wrist-injured patients. Internal consistency testing results suggested that item consistency within subset items was good and persisted over time. In conclusion, the Chinese version of the PRWE is a reliable and valid self-rated tool in measuring treatment outcome. It supplements traditional objective clinical measures and is potentially applicable in Hong Kong clinical settings. PMID- 16473731 TI - Opponens plasty rehabilitation protocol--a case report. AB - In this case report of opponens plasty, we will attempt to accomplish two objectives: 1) to characterize some innovative modifications to the standard rehabilitation protocol for an opponens plasty and 2) to explain the role and advantages of a new muscle re-education splint in this modified protocol. PMID- 16473732 TI - A biomechanical analysis of static progressive elbow flexion splinting. AB - We all have many occasions to splint the stiff elbow, especially to overcome flexion contratures, but the functional requirement of 130+degrees of elbow flexion is also an important clinical reality requiring end range splinting in no small number of cases. The author presents an analysis of the theoretical forces acting upon the elbow in two types of splints at various angles of flexion. PMID- 16473733 TI - Swing design dynamic traction splinting for the treatment of intra-articular fractures of the digits. AB - The concept of treating acute PIP fractures with traction splinting to allow early motion to mitigate against the known deleterious effects of prolonged immobilization of this problematic joint has been around for a long time and was made popular by Schenck. Our author in this PF piece has modified a splint design of Griet Van Veldhoven to further refine the dynamic traction approach. PMID- 16473734 TI - Maximizing upper extremity function: integrating our foundations, patients' goals, and evidence. Nathalie Barr lecture delivered at the ASHT Annual Meeting, San Antonio, September 2005. PMID- 16473735 TI - Building collaborative partnerships. ASHT Presidential address at the annual meeting, San Antonio, September 2005. PMID- 16473736 TI - Show me the evidence. Invited lecture from the ASHT Annual Meeting, San Antonio, September 2005. PMID- 16473737 TI - Rethinking TNM: breast cancer TNM classification for treatment decision-making and research. AB - Current classification for solid tumors is based upon characteristics of their extent. Size of the primary tumor, presence of metastatic regional lymph nodes and/or of distant metastases are the key elements for their categorization. Treatment decision-making may depend upon defined extent of disease, but it requires the knowledge of several other factors. Furthermore, effective therapeutics is less dependent upon extent of disease, biological features being increasingly instrumental for treatment choice. A new classification that integrates both requisites is proposed. The scope of this proposal is to transform the current rigid and gross categorization into a more analytical and fine tuned listing including biological variables, making staging allocation more flexible and functional for proper clinical and research needs in the present and for the future. The significant changes we propose are: PMID- 16473738 TI - Validity of needle core biopsy in the histological characterisation of mammary lesions. AB - Over the last few years, there has been an enormous increase in the use of needle core biopsy (CB) for the histopathological characterisation of suspect lesions of the breast. The aim of this study was to verify the diagnostic reliability of CB by comparing the histological results obtained with the use of this technique with those obtained from the whole of the surgically resected specimen. We studied 198 out of 426 patients with clinically and/or radiologically suspect breast lesions. We found correspondence between the histological examination of the whole of the excised specimen and that of the CB in 94.9% of the cases of infiltrating carcinoma and in 71.4% of those involving ductal carcinomas in situ. The predictive value of CB was 98.9%, sensitivity was 96.1% and specificity 93.3%. These results confirmed that CB is an extremely reliable diagnostic tool in the definition of breast lesions. PMID- 16473739 TI - Ageing, hormonal behaviour and cyclin D1 in ductal breast carcinomas. AB - Owing to the gradual modification of breast tissue in postmenopausal women, there can be differential effects on local oestrogen receptor (ER) expression, with potential impingement on the biological behaviour of cancer cells in the ageing. A series of 45 ductal carcinoma (DC) cases were selected in postmenopausal women who were not being treated with HRT. Immunohistochemical analyses were performed for hormone receptors and Ki67 expression. Fluorescence in situ hybridisation (FISH) analysis was carried out to study CCND1 amplification. The selected population was subdivided into three groups by age and was subjected to statistical studies: linear model analysis, estimation of relative incidence (RI), multivariate analysis, and nonparametric tests were performed to investigate whether there were any links between age and molecular variables in DCs. The results show a low rate of proliferation and high ER expression in the oldest age group. In the same group a close correlation was found between high ER expression and CCN in the older age group D1 amplification (P=0.000), as was a more advanced phenotype in terms of tumour size and presence of positive lymph nodes than in the other age groups considered. The results suggest that ductal breast cancer has a favourable molecular prognosis, especially in extreme old age. In particular, there is an inverse correlation between ageing and proliferation rate despite the presence of an accentuated proliferation stimulus (high ER with CCD1 amplifications) in the oldest group relative to the other groups considered. PMID- 16473740 TI - A comparison of the pattern of occurrence of breast cancer in Nigerian and British women. AB - Studies have suggested a predominance of premenopausal breast cancer in black compared to white women. The aim of the study was to compare the age specific incidence of breast cancer in Nigerian and British women. The mean age at presentation was 43.1 and 64 years for Jos (Nigeria) and Norfolk (United Kingdom), respectively. The age specific incidence rates were higher in women above 50 years compared to women less than 50 years of age in both populations. The odds of having breast cancer for women aged less than 50 years is 3.0 times higher in Norfolk (95% Confidence Interval 2.0-4.4) than Jos and 9.0 times higher for women over 50 years of age in Norfolk (95% Confidence Interval 5.3-18.3) than Jos. The age specific incidence rates are higher for postmenopausal women in both populations; with higher rates for all age groups in the United Kingdom population. PMID- 16473741 TI - Secondary breast, pancreatic, and renal involvement with non-Hodgkin's lymphoma: Imaging findings. AB - The radiologic features in a patient with breast, pancreatic, and renal non Hodgkin's lymphoma are presented in this paper. Although diagnosis of lymphoma is based on histologic criteria, radiologic techniques may be helpful in the evaluation and follow-up of masses in patients with lymphoma. PMID- 16473742 TI - Tc-99m sestamibi uptake in the chest mimicking a malignant lesion of the breast. AB - The use of nuclear medicine imaging of the breast has resulted in the improved diagnosis of breast cancer (Radiology 196 (1995) 421; J. Nucl. Med. 36 (1995) 1758). It's use is becoming more widespread, particularly as breast optimized scintigraphy with a high-resolution, breast-specific gamma camera has been developed (J. Nucl. Med. 43 (2002) 909; J. Nucl. Med. 45 (2004) 553). With the increasing use of breast-optimized scintigraphy, pitfalls in the interpretation of breast-optimized scintigraphy are being recognized. This report describes a previously unrecognized cause for a false positive interpretation of scintimammography due to tumor uptake of the radiotracer in the chest as a result of physiologic activity in the auricular aspect of the right atrium. PMID- 16473743 TI - An unusual case of breast ulceration. AB - We present an unusual case of breast ulceration secondary to heroin injection in a pregnant woman. PMID- 16473744 TI - Radiation-induced angiosarcoma of the breast shows major response to docetaxel after failure of anthracycline-based chemotherapy. AB - We report on the case of a patient with a diagnosis of an uncommon breast tumour, namely a radiation-induced angiosarcoma, which was primarily refractory to anthracycline-based chemotherapy, but highly sensitive to docetaxel. Although the sarcomas in general tend to be relatively refractory to taxanes, there is some evidence that the angiosarcomas may be sensitive to these agents. This is particularly well documented with paclitaxel, but may also be the case with docetaxel. PMID- 16473745 TI - Breast cancer multi-disciplinary teams in England: much achieved but still more to be done. AB - OBJECTIVE: To look at the national coverage, composition and discussion of breast cancer multi-disciplinary teams (MDTs) in England. DESIGN AND DATA SOURCE: All breast units in England both symptomatic and screening were sent a questionnaire. MAIN OUTCOME MEASURES: Attendance of core members at MDT meetings; Percentage of patients discussed with a treatment plan; Private patients discussed; Reconstructive surgery. RESULTS: The majority of core members of the breast MDT attend weekly meetings to discuss the multidisciplinary management of patients with breast cancer, although attendance by medical oncologists and reconstructive breast surgeons is limited. Three MDTs never had a radiologist present and 3 never had a pathologist present at the MDT meeting. Most breast MDTs have a meeting coordinator to collect case-notes, radiographs and pathology reports to facilitate the meeting. Seventy-nine out of 134 teams discuss every cancer patient and 118 also discuss private patients. Twenty-seven teams record the outcome of the MDT meeting electronically, 32 teams book surgery, 16 radiotherapy and 15 book chemotherapy direct from the MDT meeting. PMID- 16473746 TI - Is mammography overused in male patients? AB - There is no agreed protocol for the use of mammography in evaluating the male breast. As a result, the tendency is to use the mammography more often than required. In order to define the role of mammography in men, we carried out a retrospective analysis of all male patients referred to the breast clinic with a history of breast lump between January 2001 and December 2003. The impact of mammography in the evaluation of male breast cancer cases was studied. A total of 220 male patients were referred to the breast clinic during this period. Of these, 134 men had a mammographic examination, with majority (96%) being performed prior to their consultation with the breast clinician as per the clinic protocol. Nine patients under the age of 35 years also had a mammographic evaluation. There were 4 cases of breast cancer diagnosed during this period. Breast cancer was suspected in all patients on clinical examination and was confirmed by biopsy. Breast cancer in men can be suspected on clinical examination in the majority of cases. Mammography appears unnecessary in most men and should not be used as a routine imaging procedure. One should consider imaging only those with clinically suspicious breast lumps to avoid unnecessary imaging particularly in young male patients. PMID- 16473747 TI - Complementary or alternative medicine: towards establishing 'value for money'. PMID- 16473748 TI - The effect of topical wheatgrass cream on chronic plantar fasciitis: a randomized, double-blind, placebo-controlled trial. AB - OBJECTIVES: To investigate the efficacy of a topical wheatgrass cream for improving pain and function in patients with chronic plantar fasciitis. DESIGN: Randomized, double-blind, placebo-controlled trial. SETTING: Eighty participants with chronic plantar fasciitis were randomly assigned to a treatment group (wheatgrass cream) or a control group (placebo cream). All participants applied a cream twice daily for 6 weeks. Follow up was conducted at 6 and 12 weeks. MAIN OUTCOME MEASURES: Visual Analogue Scale (VAS) for daily first-step pain and the Foot Health Status Questionnaire (FHSQ) for overall foot function. Secondary measures of foot posture, calf muscle strength and range of ankle dorsiflexion were also assessed. RESULTS: No significant differences were found between groups with respect to main outcomes of first-step pain or foot function at any time. Both groups improved significantly from baseline to 6 weeks, and these improvements were maintained at 12 weeks. CONCLUSIONS: The topical application of wheatgrass cream is no more effective than a placebo cream for the treatment of chronic plantar fasciitis. PMID- 16473749 TI - Patient requests and provider suggestions for alternative treatments as reported by rural and urban care providers. AB - OBJECTIVES: Explored the relationship between different types of care providers' willingness to suggest alternative and complementary treatments (CAM), patients' requests for CAM, and provider perceptions about CAM as barriers to effective healthcare. DESIGN: Large survey. SETTING: Alaska and New Mexico. MAIN MEASURES: Survey responses from 1528 physical and behavioral healthcare providers. RESULTS: Over 97% of providers suggested CAM; over 97% reported patients asked for CAM. Providers were more likely to suggest CAM than perceived CAM as a barrier to care. Healthcare providers who were female, from small rural areas, or specializing in behavioral healthcare were more likely to suggest CAM and less likely to perceive CAM as a barrier. Patients of physical healthcare providers asked for CAM more often than patients of behavioral healthcare providers, yet physical care providers suggested CAM less frequently. CONCLUSIONS: Healthcare providers of all disciplines, regions, and gender are sensitive to patients' desire for CAM and do not perceive CAM as a barrier to care. PMID- 16473750 TI - Patients seeking care from acupuncture practitioners in the UK: a national survey. AB - OBJECTIVE: The primary aim was to describe the characteristics of acupuncture patients and in particular the main problem or symptom for which they were seeking treatment. Our secondary aim was to compare the profiles of acupuncture patients with those of a patient survey undertaken in 1988. METHODS: We used the data from a recent prospective adverse event survey of a representative sample of 9408 acupuncture patients who were consulting members of the British Acupuncture Council. We analysed patient reports of demographic details, pathways to care, whether the National Health Service paid for their treatment, whether they had previously consulted a GP or hospital specialist for their main problem and whether they were consulting an acupuncturist for the first time. We separately analysed equivalent data from a survey funded by the Nuffield Provincial Hospital Trust undertaken in 1988. RESULTS: Seventy-four percent of patients were female, and with an average age of 51 years. Most commonly, patients had self-referred (39%), had previously consulted their doctor about their problem or symptom (78%), were paying for their own treatment (95%), and had received acupuncture before (87%). The most common main problem or symptom reported by patients was musculo-skeletal (38%), followed by psychological (11%), general (9%), neurological (8%) and gynaecological/obstetric (8%), while 5% of patients were seeking treatment for their general well-being. In comparing our data with that of the 1988 survey, we found a number of changes over time, in particular the proportion of musculo-skeletal conditions had dropped significantly (chi(2) = 17.36, d.f. = 1, p < 0.001), indicating a wider case mix amongst patients compared with those seeking care in 1988. CONCLUSION: This evidence from acupuncture patients' reports shows that musculo-skeletal problems provide the main reason for seeking treatment. The large dataset from this study provides a wealth of information and a fresh raft of questions which will inform future research and policy-making. PMID- 16473751 TI - In vivo dc and ac measurements at acupuncture points in healthy and unhealthy people. AB - OBJECTIVES: The aim of this work was to compare in vivo measurements of direct current (dc) and alternating current (ac) obtained from acupuncture points in Ukrainian and Mexican residents. METHODS: Measurements were made using the method of Voll. The participants were 43 healthy Ukrainian and 71 healthy Mexican residents aged between 20 and 30 years, as well as 24 Mexican patients with a clinical diagnosis of rheumatoid arthritis and 14 patients with a clinical diagnosis of allergy. RESULTS: The results showed that dc measurements are not directly applicable to different populations. Thus, the dc resistance of the acupuncture points in the Mexican participants was 4-5 times larger than in the Ukrainians. In contrast, the capacitance of the two groups did not differ by more than 25%. CONCLUSIONS: Impedance measurements from acupuncture points can be used as an efficient and prompt non-invasive method for diagnostic purposes. PMID- 16473752 TI - Acupuncture for tinnitus: a series of six n = 1 controlled trials. AB - OBJECTIVE: To explore patient perceived benefits of acupuncture for tinnitus. DESIGN: Controlled n=1 trials, with two phases A and B. SUBJECTS: Six patients with tinnitus. OUTCOME MEASURES: Primary outcome was Daily Diary records related to four tinnitus symptoms: loudness of tinnitus; pitch of tinnitus; waking hours affected with tinnitus; quality of sleep. Secondary outcomes were the Tinnitus Handicap Inventory (THI) and Measure Your Medical Outcome Profile (MYMOP). METHODS: Patients received a course of 10 acupuncture treatments over a 2-week period. Daily Diary entries related to the four tinnitus symptoms were recorded by patients for 14 days pre-treatment (phase A) and 14 days post-treatment (phase B). A hierarchical Bayesian model was used to combine the results from the individual patients to obtain estimates of the population and individual patient treatment effects, incorporating random variations at both levels (between patients and within patient). Tinnitus Handicap Inventory (THI) and Measure Your Medical Outcome Profile (MYMOP) were recorded at assessment points pre-treatment and post-treatment. RESULTS: Six patients participated in the trials, each receiving 10 treatments and completing all Daily Diary entries and outcome measures. For the of symptoms of loudness and pitch, there were variable treatment effects between patients, with a trend for the median overall reduction for loudness of -2.49 (-5.04, 0.02) and for pitch -1.39 (-3.74, 0.89), 95% credibility intervals being shown in brackets. For the other two symptoms, the waking hours affected and quality of sleep, patients' responses were more consistent, with amore credible overall median reduction for affected waking hours of -2.76 (-3.94, -1.63) and for quality of sleep -2.72 (-3.45, -2.03). The THI and MYMOP measures showed a trend of improvement after treatment. CONCLUSION: The n=1 trial methodology, with an AB design and Bayesian analysis, can be considered of value in exploring treatment effects for small numbers of patients receiving individualised treatments, as is common within complementary medicine. When the treatment effects from six patients were synthesized, the results of this study suggest that acupuncture may have a beneficial role in the treatment of tinnitus. PMID- 16473753 TI - The therapeutic efficacy of somatic acupuncture is not increased by auriculotherapy: a randomised, blind control study in cervical myofascial pain. AB - Auriculotherapy (ear acupuncture) is a therapeutic technique in which points on the auricle are stimulated with needles. Usually it is combined with somatic acupuncture because of possible synergy, although the efficacy of this pairing has neither been confirmed nor disproved. The aim of this study was to verify: (1) if somatic acupuncture can reduce myofascial cervical pain; (2) if concomitant auriculotherapy improves the efficacy of somatic acupuncture. A group of 62 patients affected by cervical myofascial pain was randomly divided into two groups of 31. Group A (6 males and 25 females) underwent eight sessions of somatic acupuncture. Group B (7 males and 24 females) underwent eight sessions of somatic acupuncture in the same way as group A, paired with auriculotherapy. Pain was scored using the McGill Pain Questionnaire before and at the end of treatment, and 1 and 3 months later. The results showed that both somatic acupuncture and somatic plus ear acupuncture have a positive effect in reducing pain. The pain intensity score was 40.70 +/- 17.78 in group A before therapy and 13.32 +/- 9.62 after therapy; in group B it was 38.90 +/- 15.31 and 13.43 +/- 10.96. Somatic plus auriculotherapy was therefore not statistically significantly superior to somatic therapy alone in the treatment of cervical myofascial pain. PMID- 16473754 TI - A novel sham moxibustion device: a randomized, placebo-controlled trial. AB - OBJECTIVES: (1) Develop a sham moxibustion device; (2) determine whether volunteer participants and practitioners can distinguish the sham procedure from real moxibustion during treatment for prevention of the common cold; and (3) assess the feasibility of conducting a larger clinical trial of the device. DESIGN: Double-blinded, randomized, placebo-controlled clinical trial. SETTING: A community outpatient health station in Chaoyang district, Beijing, China, from early November through late December 2003. METHODS: Sham and real moxibustion pillars were made by affixing moxa cones onto special collar bases. These pillars resemble each other in appearance, burning procedure and residue, but the base of the sham pillar isolates the moxa-produced heat and smoke and prevents them from radiating to the skin. These devices were tested in a pilot clinical trial in which patients received moxibustion at Zusanli (ST 36) for prevention of the common cold. Volunteers (n = 71) aged 55-75 years were given pre-treatment questionnaires to assess their knowledge of moxibustion and their expectations and motives for participating in the trial, randomized into treatment (n = 36) and placebo-controlled (n = 35) groups, and treated once every 2 days for 1 month. Questionnaires assessing the effectiveness of the blinding were given to the volunteers at the end of the trial and to practitioners after each treatment session. RESULT: There were no significant differences between the two groups of patients in past moxibustion experience, knowledge of moxibustion, expectations and motivation. The number of volunteers in the treatment group and the placebo group who believed they received: (1) real moxibustion; (2) sham moxibustion; and (3) were uncertain as to which they received is 30, 0, 5; 29, 0, 4, respectively. The results show no significant statistical differences (P = 1.000). The number of volunteers in the treatment group and the placebo group who were believed, by the two practitioners who provided the treatments, to have received: (1) real moxibustion; (2) sham moxibustion; and (3) and an undisclosed treatment were 10, 6, 19; 9, 4, 20 and 11, 7, 17; 12, 5, 16, respectively. There were no statistically significant differences between the practitioners (P = 0.811 and 0.840). CONCLUSION: The sham moxibustion device was successfully validated in the present study. The results demonstrate that these specially designed real and sham moxibustion pillars can be used together to provide an effective placebo control in moxibustion research and can successfully fulfill the double blinding protocol in moxibustion clinical trials. PMID- 16473756 TI - Topical and oral CAM in acne: a review of the empirical evidence and a consideration of its context. AB - INTRODUCTION: Acne is one of the commonest diseases to afflict humanity. Anecdotally, the use of CAM in acne is widespread. In this review the empirical evidence for the efficacy of CAM modalities is examined and the context for their use discussed. METHODS: Searches were made of the MEDLINE, EMBASE, AMED (Allied and Complementary Medicines), Cochrane, and DARE databases using the search terms "acne" and "acne vulgaris" together with "alternative" and "complementary". RESULTS: The identified studies examined a broad range of CAM modalities but were of generally poor methodological quality. Evidence suggests that many of these therapies are biologically plausible. DISCUSSION AND CONCLUSION: Complementary therapies in acne should be viewed in a wider context than that of the very limited empiric evidence base that exists for their use. Further rigorously conducted trials should be conducted to define efficacy and adverse effect profiles of currently used CAM acne therapies. PMID- 16473757 TI - Documentary analysis in CAM research: Part 2. AB - The analysis of documents can provide insights into important social and political issues related to complementary and alternative medicine (CAM). This paper identifies some important sources of documents and outlines some of the advantages and disadvantages of using documents in social research. A number of analytic strategies are illustrated with examples to demonstrate the potential for documentary analysis to answer particular CAM-related research questions. These strategies are described as quantitative content analysis, qualitative descriptive analysis, qualitative interpretive analysis and semiotic analysis. It is argued that documents are a convenient source of data and that their analysis can enrich our understanding of CAM. Part 1 covered quantitative and qualitative strategies. Part 2 focuses on semiotic analysis and the methodological strengths and weaknesses of the different strategies. PMID- 16473758 TI - Recruitment and implementation strategies in randomised controlled trials of acupuncture and herbal medicine in women's health. AB - INTRODUCTION: Recruitment of subjects to clinical trials is known to be difficult and there is little research describing recruitment and implementation strategies to CAM clinical trials. This paper describes the experiences from recruitment and implementation for five randomised controlled trials in women's health conducted in South Australia. METHODS: Descriptive study reporting on trial documentation and questionnaires completed by women. RESULTS: Recruitment rates varied between trials and a variety of recruitment strategies were used between studies. Promotion of the trials to the wider community were facilitated by issuing press releases and subsequent reporting by the media. Women found the trial acceptable suggesting factors motivating and preventing women from participating were well addressed. CONCLUSION: It is hoped that the experiences described here will give some insight to recruitment and implementation strategies. There is a need for more systematic research and evaluation of these strategies, and dissemination of these findings to assist with successful implementation of trials. PMID- 16473759 TI - Integration--a key to success in the genetic dissection of complex diseases? AB - Complex diseases are polygenic and multifactorial. The outcome of two decades of search for the culprit genes in complex diseases involving the cardiovascular system has been less than satisfactory. Genomic studies using linkage analysis have led so far to the detection of a large number of quantitative trait loci that embed a large number of candidate genes. Transcriptomic studies using differential gene expression profiling and DNA microarrays have also generated hundreds of potential candidate genes. None of these genetic strategies has enabled researchers to reduce the number of genes to a manageable number or to identify the specific culprit genes. We recently proposed that the search for genes involved in complex diseases such as hypertension might benefit from an integration of genomics and transcriptomics as a logical alternative strategy to using either approach alone. We applied this integrated genomic-transcriptomic approach to identify the genes that are involved in the pathogenesis of hypertension in the Sabra rat model of salt susceptibility. We successfully identified seven novel candidate genes for hypertension, an outcome that could not have been achieved by genomics or transcriptomics alone. PMID- 16473760 TI - Vang-like 2 and noncanonical Wnt signaling in outflow tract development. AB - Despite rapid advances in cardiovascular developmental genetics, the precise morphogenetic processes that coordinate heart development, and the genes and signaling pathways that regulate them remain unclear. In this review, we describe a highly conserved signaling pathway, the noncanonical Wnt (planar cell polarity) pathway, and its relationship to cardiovascular development and congenital heart defects. This pathway regulates cell polarity and polarized cell movements in a variety of contexts. Mutations in several genes in this pathway and specifically in the Vang-like 2 (Vangl2) (strabismus) gene, result in abnormalities in the remodeling of the outflow tract and, ultimately, in the cardiac alignment defect double-outlet right ventricle. Polarized cell migration of cardiomyocytes into the outflow tract cushions is inhibited when Vangl2 function is disturbed, suggesting that the noncanonical Wnt pathway may regulate this aspect of outflow tract remodeling. These studies suggest that mutations in Vangl2 and other components of the noncanonical Wnt pathway, may be candidates for causing congenital outflow tract defects in humans. PMID- 16473761 TI - Definitive hematopoiesis from endothelial cells in the mouse embryo; a simple guide. AB - Circulation is composed of two interactive systems, the cardiovascular and the hematopoietic, which affect each other. Recently, endothelial progenitor cells/angioblasts have been identified in the circulation of the adult mouse and human. Furthermore, some hematopoietic cells (HCs) have been shown to contribute to angiogenesis, suggesting that HCs can transdifferentiate into endothelial cells (ECs). Although these concepts in adult are still controversial, understanding the mechanisms of the relationship between ECs and HCs would benefit the clinical application for cardiovascular and hematologic disorders. Both ECs and HCs are considered to be derived from a common germ layer, the mesoderm, and have more intimate relationship in embryo than in adult. Here, we describe the relationship between ECs and HCs with special attention to the hemogenic ECs in the mouse embryo. PMID- 16473762 TI - Caveolin-1: dual role for proliferation of vascular smooth muscle cells. AB - Although caveolae function in vesicular and cholesterol trafficking, the recent identification of various signaling molecules in caveolae and their functional interaction with caveolin suggest that they may participate in transmembrane signaling. Interestingly, many of the signaling molecules that interact with caveolin-1 (cav-1) mediate mitogenic signals to the nucleus, implying that cav-1 may play a modulating role in the pathophysiology of vascular proliferative diseases such as atherosclerosis and restenosis after angioplasty. Although much attention has been given to the predominantly antiproliferative role of cav-1 in growth-factor-induced signal transduction, we were recently able to demonstrate that cav-1 acts in mechanotransduction too. During cyclic strain, however, cav-1 is critically involved in proproliferative signaling. We propose that, at least in the vasculature which is constantly exposed to alternating mechanical force and different growth factors, cav-1 holds a dual role toward modulation of proliferation, depending on the stimulus the cells are exposed to. In vivo, the net effect of growth factors and mechanically triggered stimuli determines the amount of local cell proliferation and, therefore, the onset and progression of vascular proliferative disease. PMID- 16473764 TI - Atherosclerosis and the Lectin-like OXidized low-density lipoprotein scavenger receptor. AB - The Lectin-like OXidized low-density lipoprotein scavenger receptor (LOX-1) is implicated in vascular inflammation and atherosclerotic plaque initiation, progression, and destabilization. LOX-1 levels are elevated upon recognition of oxidized low-density lipoprotein, a key pro-atherogenic substance in the vasculature. Recent evidence indicates this gene product is a biomarker of inflammation and disease status. We review and assess the role of LOX-1 in atherosclerotic plaque formation, physiologic regulation, and as a biomarker and target in cardiovascular disease diagnosis and prevention. PMID- 16473763 TI - Angiogenesis: the VE-cadherin switch. AB - Because angiogenesis is a key step in a number of pathologic processes, including tumor growth and atherosclerosis, many research studies have investigated the regulatory signals active at various stages of vascular invasion. The differential activities of the endothelial junction protein vascular endothelial (VE)-cadherin reflect the versatile behavior of endothelial cells between vascular quiescence and angiogenesis. VE-cadherin function and signaling are deeply modified in proliferating cells, and this conversion is accompanied by phosphorylation of the protein on tyrosine residues and enhanced transcription of its gene. Recent advances in the complex interplay between protein tyrosine kinases and phosphatases regulating VE-cadherin phosphorylation and function are discussed in this review. PMID- 16473765 TI - Prevention of infections by probiotics. AB - 'Viable bacteria that exhibit beneficial effects for health based on improvement of balance of intestinal bacterial flora' was the most common definition of probiotics, but the diversity of their action mechanisms, including immune regulation, has been clarified, and probiotics have recently been broadly defined as 'live microorganisms which when administered in adequet amounts confer a health benefit on the host'. Coined words such as prebiotics, describing non digestible food fiber components that contribute to host health by activating proliferation and function of beneficial intestinal bacteria, and synbiotics, describing a combination of probiotics and prebiotics have also been established as medical expressions. In this report, clinical studies of probiotics, prebiotics, and synbiotics on prevention of infection by various infectious diseases in humans are reviewed. The effects of and action mechanisms against sporadic intestinal infectious diseases that are difficult to investigate in humans (enterohemorrhagic Escherichia coli and Salmonellosis) in experimental animal models are also reviewed. Finally, points necessary for clarification of the role of probiotics in health care, and their functions in health care foods are discussed. PMID- 16473766 TI - Biosynthetic regulation of phytochelatins, heavy metal-binding peptides. AB - Phytochelatins (PCs) are heavy metal-binding peptides that play important roles in the detoxification of toxic heavy metals and the regulation of intracellular concentrations of essential metals in eukaryotes, including higher plants, fungi, and microalgae. Recently, PC synthase genes in higher plants and fission yeast have been identified and characterized, enabling molecular biological studies to unravel the mechanisms underlying PC synthesis. Moreover, recent routine database searches have unexpectedly identified genes that are similar to plant PC synthase genes in the genomes of worms and some prokaryotes. In this review, we introduce these recent advances in our understanding of the molecular mechanisms for PC biosynthesis and functions in order to supply basic information about the unique and attractive peptides applicable to various fields. PMID- 16473767 TI - Potential of predominant activated sludge bacteria as recipients in conjugative plasmid transfer. AB - We investigated the possibility of conjugative plasmid transfer to the predominant bacteria in activated sludge and the factors influencing the transfer frequency in the activated sludge process. We performed conjugative transfers of a self-transmissible, broad-host-range plasmid RP4 from Escherichia coli C600 to activated sludge bacteria by broth mating. Most of the activated sludge bacteria tested could acquire plasmid RP4, although the transfer frequencies varied from 8.8 x 10(-7) to 1.3 x 10(-2) transconjugants per recipient. The transfer frequencies in several strains were similar to, or higher than, that in intraspecific transfer to E. coli HB101. Matings under various environmental conditions showed that factors relevant to physiological activity, such as temperature and nutrient conditions, seemed to affect the transfer frequency. In addition, conjugative transfer was detected even in filtered raw and treated wastewaters. Thus, the predominant activated sludge bacteria seem to have sufficient potential as recipients in conjugative plasmid transfer under the conditions likely to occur in the activated sludge process. Transfer frequency was reduced by agitation in the presence of suspended solid. This may suggest that conjugative plasmid transfer is physically inhibited in aeration tanks. PMID- 16473768 TI - Production of L-lactic acid from fresh cassava roots slurried with tofu liquid waste by Streptococcus bovis. AB - To reduce the production cost of biodegradable plastics, the fermentation performance of L-lactic acid for a new fermentation medium, fresh cassava roots (FCRs) as a substrate slurried with tofu liquid waste (TLW) as basal medium, was investigated by batch fermentation of Streptococcus bovis. The fermentation properties of the three substrates, namely, FCR, tapioca (cassava starch) and glucose, which were independently mixed with TLW, were compared with those independently mixed with the standard basal medium, trypto-soya broth (TSB). Experiments were conducted at various sugar concentrations of the substrates with CaCO(3) as a neutralizer. The maximum L-lactic acid concentrations (C(La)) obtained using the three substrates in TLW were about 75% of those obtained using TSB caused by less nutrients in the TLW. The L-lactic acid productivities (P(La)) and the specific growth rates of S. bovis (mu) in TLW were about 1/4 to 1/3 and 1/5 to 1/4 of those in TSB, respectively. The maximum C(La), P(La) and mu were obtained at 10% w/w sugar concentration. Total yields (eta) were nearly constant up to 10% w/w sugar concentration for TSB and TLW, that is, 80% to 85% and 50% to 60%, respectively. But their total yields decreased in more than 10% w/w sugar concentration in both basal media, because of substrate inhibition. The fermentation properties (C(La), P(La), mu, and eta) were found to be in the order of: FCR > tapioca > glucose for all concentrations of the three substrates. The fermentation properties for FCR and tapioca were higher than those for glucose, in TLW or TSB, because S. bovis in a medium containing starch (FCR and tapioca) has more amylase activity than in a medium containing glucose. The nutrients in FCR with poor nutrient basal medium (TLW) more strongly affected the fermentation properties than those in FCR with rich nutrient basal medium (TSB). The proposed fermentation medium of FCR slurried with TLW is worth studying in order to reduce production cost of biodegradable plastics. PMID- 16473769 TI - High hydrostatic pressure treatment impairs AcrAB-TolC pump resulting in differential loss of deoxycholate tolerance in Escherichia coli. AB - We reported previously that high hydrostatic pressure-injured stationary phase cells of Escherichia coli K-12 lost their intrinsic deoxycholate tolerance. The AcrAB-TolC multi-drug resistance pump driven by proton motive force has been argued to be responsible for the tolerance to deoxycholate. In this report, we tested the sensitivity of the AcrAB-TolC (three components) pump to high hydrostatic pressure treatment (HPT). E. coli K-12 treated with HPT became sensitive to AcrAB-TolC-specific drugs such as ethidium bromide, but not to tetracycline which is pumped out by a one-component transporter, Tet. Only E. coli K-12 overproducing both AcrAB and TolC exhibited restored tolerance to deoxycholate after HPT but not E. coli overproducing either TolC or AcrAB. These observations strongly suggest that three-component pumps such as AcrAB-TolC are more susceptible to HPT than one-component pumps such as Tet, resulting in the differential loss of deoxycholate tolerance in high hydrostatic pressure-injured E. coli cells. PMID- 16473770 TI - Transformation of oil-producing fungus, Mortierella alpina 1S-4, using Zeocin, and application to arachidonic acid production. AB - The arachidonic acid-producing fungus Mortierella alpina 1S-4, an industrial strain, was endowed with Zeocin resistance by integration of the Zeocin resistance gene at the rDNA locus of genomic DNA. Plasmid DNA was introduced into spores by microprojectile bombardment. Twenty mg/ml Zeocin completely inhibited the germination of M. alpina 1S-4 spores, and decreased the growth rate of fungal filaments to some extent. It was suggested that preincubation period and temperature had a great influence on transformation efficiency. Four out of 26 isolated transformants were selected. Molecular analysis of these stable transformants showed that the plasmid DNA was integrated into the rDNA locus of the genomic DNA. We expect that this system will be applied for useful oil production by gene manipulation of M. alpina 1S-4 and its derivative mutants. On the basis of the fundamental transformation system, we also tried to overexpress a homologous polyunsaturated fatty acid elongase gene, which has been reported to be included in the rate-limiting step for arachidonic acid production, thereby leading to increased arachidonic acid production. PMID- 16473771 TI - Purification and properties of an extracellular endo-1,4-beta-xylanase from Penicillium citrinum and characterization of the encoding gene. AB - An extracellular endo-1,4-beta-xylanase was purified from the culture filtrate of a filamentous fungus Penicillium citrinum FERM P-15944 grown on birch-wood xylan. The purified enzyme showed a single band on SDS-PAGE with an apparent M(r) of 20,000 and had an isoelectric point below 3.5. Xylanase activity was optimal at pH 5.0 and 55 degrees C. The genomic DNA and cDNAs encoding this protein were cloned and sequenced. Southern blot analysis indicated that the xylanase gene (xynA) was present as a single copy in the genome. An open reading frame of 657 bp was interrupted by two introns of 65 and 55 bp, and encoded a presumed prepropeptide of 27 amino acids and a mature protein of 190 amino acids. Three distinct transcription start points were observed at positions -20 (A), -31 (A), and -36 (A) from the start codon. The 5'-noncoding region had a putative TATA box at nt -66 (TATAAA). The xynA cDNA was functionally expressed under the control of the alcohol oxidase I gene promoter in the methylotrophic yeast Pichia pastoris. A neighbor-joining tree showed that the P. citrinum enzyme is closely related to several other fungal xylanases belonging to the glycoside hydrolase family 11: Trichoderma reesei XYN2, Aspergillus niger xynNB, Penicillium funiculosum xynC, Penicillium sp. strain 40 xynA, Chaetomium gracile cgxB, and Aspergillus nidulans xlnA and xlnB. PMID- 16473772 TI - Extracellular trypsin-like proteases produced by Cordyceps militaris. AB - A trypsin-like protease, P-1-1, was purified from the culture supernatant of the fungus Cordyceps militaris by (NH(4))(2)SO(4) precipitation, chromatography on DEAE Bio-Gel Agarose, TSKgel CM-5PW, and gel-filtration with HiLoad 26/60 Superdex 75 pg, and its properties were examined. Purified P-1-1 showed a single band by SDS-PAGE and was estimated to have a molecular mass of 23,405 by matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS). The optimum pH of the enzyme was between 8.5 and 12.0. It was inhibited strongly by leupeptin and diisopropyl fluorophosphate (DFP), and definitely did by N(alpha)-tosyl-L lysine chloromethyl ketone hydrochloride (TLCK), phenylmethanesulfonyl fluoride (PMSF) and chymostatin. The carbonyl group sides of Arg and Lys were confirmed as the sites of cleavage by the enzyme toward cecropin B. These results indicate that P-1-1 is a trypsin-type serine protease. The N-terminal amino acid sequence of P-1-1 showed a high homology with those of trypsins or chymotrypsin derived from Diptera insects. PMID- 16473773 TI - Evaluation of pretreatment with Pleurotus ostreatus for enzymatic hydrolysis of rice straw. AB - The effects of biological pretreatment of rice straw using four white-rot fungi (Phanerochaete chrysosporium, Trametes versicolor, Ceriporiopsis subvermispora, and Pleurotus ostreatus) were evaluated on the basis of quantitative and structural changes in the components of the pretreated rice straw as well as susceptibility to enzymatic hydrolysis. Of these white-rot fungi, P. ostreatus selectively degraded the lignin fraction of rice straw rather than the holocellulose component. When rice straw (water content of 60%) was pretreated with P. ostreatus for 60 d, the total weight loss and the degree of Klason lignin degraded were 25% and 41%, respectively. After the pretreatment, the residual amounts of cellulose and hemicellulose were 83% and 52% of those in untreated rice straw, respectively. By enzymatic hydrolysis with a commercial cellulase preparation for 48 h, 52% holocellulose and 44% cellulose in the pretreated rice straw were solubilized. The net sugar yields based on the amounts of holocellulose and cellulose of untreated rice straw were 33% for total soluble sugar from holocellulose and 32% for glucose from cellulose. The SEM observations showed that the increase in susceptibility of rice straw to enzymatic hydrolysis by pretreatment with P. ostreatus is caused by partial degradation of the lignin seal. When the content of Klason lignin was less than 15% of the total weight of the pretreated straw, enhanced degrees of enzymatic solubilization of holocellulose and cellulose fractions were observed as the content of Klason lignin decreased. PMID- 16473774 TI - Diversity of 16S ribosomal DNA-defined bacterial population in acid rock drainage from Japanese pyrite mine. AB - Four acidophilic bacteria (YARDs1-4) were isolated from an acid rock drainage (ARD) from Yanahara mine, Okayama prefecture, Japan. The physiological and 16S rDNA sequence analyses revealed that YARD1 was closely affiliated with Acidithiobacillus ferrooxidans, YARD2 was an Acidiphilium-like bacterium, and YARD3 and YARD4 were sulfur-oxidizing bacteria with a relatively close relationship to A. ferrooxidans in the phylogenetic analysis. A molecular approach based on the construction of a 16S rDNA clone library was used to investigate the microbial population of the ARD. Small-subunit rRNA genes were PCR amplified, subsequently cloned and screened for variation by a restriction fragment length polymorphism (RFLP) analysis. A total of 284 clones were grouped into 133 operational taxonomic units (OTUs) by the RFLP analysis. Among them, an OTU showing the same RFLP pattern as those of the isolates from the ARD was not detected. The phylogenetic analysis based on the 16S rDNA sequences from 10 major OTUs and their close relatives revealed that 4 OTUs containing 32.1% of the total clones were loosely affiliated with Verrucomicrobia, 2 OTUs containing 6.6% of the total clones were loosely affiliated with Chloribi, and other OTUs were affiliated with Actinobacteria, Nitrospirae, and beta-Proteobacteria. PMID- 16473775 TI - Horseradish peroxidase degrades lipid hydroperoxides and suppresses lipid peroxidation of polyunsaturated fatty acids in the presence of phenolic antioxidants. AB - Linoleic acid hydroperoxide (LAOOH) was effectively degraded by horseradish peroxidase (HRP) in the presence of quercetin. Several natural phenolic antioxidants, such as quercetin, capsaicin, and alpha-tocopherol, acted as good hydrogen donors in the peroxidase reaction that occurs during lipid hydroperoxide degradation. However, glutathione, which is a non-phenolic antioxidant that acts as a hydrogen donor for glutathione peroxidase, could not suppress lipid peroxidation in the presence of HRP. Lipid hydroperoxides generated from eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were also degraded with HRP in the presence of quercetin, and oxidative decomposition of DHA was suppressed by this reaction. PMID- 16473777 TI - Enzymatic chemoselective synthesis of secondary-amide surfactant from N methylethanol amine. AB - Efficient selective synthesis of the secondary amide surfactant N-methyl lauroylethanolamide from methyl laurate and N-methylethanol amine by carrier fixed Chirazyme L-2 (Candida antarctica) using a kinetic strategy has been demonstrated. When different solvents were screened for product yields using Chirazyme L-2, acetonitrile was found to be optimal. The rate of the reaction increased sharply by increasing the molar ratio of the reactants and the reaction temperature. When the reaction was performed at 50 degrees C for 36 h with 50 mmol ester and 100 mmol amine, the product was obtained in a 97.1% yield. With 50 mmol ester and 150 mmol amine, the highest yield (97.3%) was obtained after 16 h of incubation at 50 degrees C. It took only 5 h to get a yield of 95.8% at 60 degrees C using 50 mmol ester and 200 mmol amine. The enzyme activity in the amidation reaction mixture did not decrease notably even after six uses. PMID- 16473776 TI - Enhancement of cometabolic biodegradation of trichloroethylene (TCE) gas in biofiltration. AB - A biofilter column inoculated with Pseudomonas putida F1 was operated to study cometabolic biodegradation of trichloroethylene (TCE) gas using toluene as a primary substrate. Variations in the efficiency and capacity of TCE elimination with different inlet concentrations of toluene and TCE were investigated in order to understand the competitive inhibition between toluene and TCE. Two toluene feeding methods, stage feeding along the column and cyclic feeding, were examined as strategies to enhance TCE cometabolic biodegradation by avoiding the toluene inhibition of TCE biodegradation and the toxic effect of TCE on cells and toluene dioxygenase enzymes. It was concluded that both methods are promising and that the determination of a suitable feeding frequency, recovery period, and inlet toluene concentration was required to optimize cyclic feeding in the cometabolic biodegradation of TCE. PMID- 16473778 TI - Preparation of silk protein sericin as mitogenic factor for better mammalian cell culture. AB - We previously reported that sericin small (sericin-S), with a molecular weight that ranges from 5 to 100 kDa, is a cell culture supplement used to accelerate cell proliferation. In this study, a novel preparation method for sericin and several applications of sericin were examined. Sericin large, prepared under nonhydrolyzing conditions and ranging from 50 to 200 kDa, also accelerated cell proliferation, but its effects were inferior to those of sericin-S. Additional sericin preparations with various molecular weights that were differentially hydrolyzed were also tested but none of them was significantly superior to sericin-S, and neither were several recombinant sericin peptides. Sericin-S successfully accelerated the proliferation of hybridoma cells in various serum free media, implying the mitogenic effect of sericin is independent from media. We also demonstrated that sericin-S successfully induced the proliferation of CTLL-2, an established T lymphocyte cell line, under IL-2 starvation conditions. These results indicate that sericin, particularly sericin-S, improves serum-free mammalian cell culture. PMID- 16473779 TI - Improvement of growth stability of photosynthetic bacterium Rhodobacter capsulatus. AB - Using a semicontinuous culture method, in which operational parameters such as cell concentration and light intensity distribution were maintained almost constant, instability of the specific growth rate of Rhodobacter capsulatus B 100, a purple bacterium, was observed to be similar to that of R. capsulatus ST 410 when cultivated under high ratios of light intensity on the illuminated side to that of the transmitted light. Such instability was not observed in the cultivation of Chlorella vulgaris, a eukaryotic green alga, even at higher cell concentrations. Under the same conditions, the increase in only the ferrous concentration from 43 microM, the concentration in the original RCV medium, to 172 microM sustained a stable growth, whereas Fe(2+) was slightly consumed during the cultivation. Supplemental illumination with a fluorescent lamp on the transmitted side of a flat plate photobioreactor sustained a moderate level of stable growth, while a halogen lamp slightly affected the growth stability. Our results showed that an increase in Fe(2+) concentration or supplemental illumination improves the growth stability of R. capsulatus. PMID- 16473780 TI - Amplified fragment length polymorphism of the AWA1 gene of sake yeasts for identification of sake yeast strains. AB - Sake yeasts are used for sake brewing and have a crucial role in the quality of sake, since they produce not only ethanol but also various compounds that provide sake flavors. Therefore, the appropriate selection and monitoring of a strain used in sake mash is important. However, the identification of specific sake yeast strains has been difficult, because sake yeasts have similar characteristics in taxonomic and physiological analyses. We found amplified fragment length polymorphisms (AFLPs) in the PCR products of the AWA1 gene of sake yeast strains. The AWA1 gene encodes a cell wall protein that is responsible for foam formation in sake mash. This polymorphism of the AWA1 gene can be used for the identification of sake yeast strains. PMID- 16473781 TI - Ca(2+)-dependent cell aggregation of halophilic archaeon, Halobacterium salinarum. AB - Halobacterium salinarum CCM 2090 exhibits a Ca(2+)-dependent cellular aggregation system. This process is caused by the interaction between Ca(2+) and aggregation factor(s). Although the aggregates formed are encased in matrix-like substances, such as bacterial biofilms, they are not attached to the abiotic substratum. Therefore, these substances may be distinguished from typical biofilms. PMID- 16473782 TI - Analysis of isoflavone content in tempeh, a fermented soybean, and preparation of a new isoflavone-enriched tempeh. AB - To produce a tempeh-like functional food containing a high level of isoflavone with a high absorptivity, we analyzed changes in the composition of isoflavone during tempeh fermentation and the difference in isoflavone content depending on the soybean variety and particular tissue. By adding soybean germ (hypocotyl) that contained a large amount of isoflavone, we prepared a new isoflavone enriched tempeh in the form of a granular fermented soybean-based food, which can serve as a nutritious supplement for the elderly. PMID- 16473784 TI - ICD therapy: what have we learned from the clinical trials? AB - Development of implantable cardioverter defibrillators (ICD) has been a dramatic advancement in the management of patients with life-threatening ventricular arrhythmias. We hereby reviewed the landmark clinical trials on ICD with special emphasis on late-breaking clinical trials and assessed their impact on every-day decision making and patient selection for ICD implantation. PMID- 16473785 TI - Transcatheter closure of muscular ventricular septal defect using the Amplatzer devices. AB - TECHNIQUE: From November 1997 to June 2002, percutaneous transcatheter closure of muscular ventricular septal defects was attempted in seven patients. Four patients had single and three had multiple defects. Surgical closure was performed in two patients in an attempt to close a perimembranous defect, leaving behind a large apical muscular defect, which was successfully closed using a device in one patient, whilst the second patient succumbed to septicemia/endocarditis 3 weeks after failure of device implantation. One patient had previous pulmonary artery banding and in another intraoperative placement of two Clamshell devices followed by additional transcatheter closure using Gianturco coils in two different sessions was performed. RESULTS: Transcatheter closure of ventricular septal defect was successful in six patients. Three patients underwent closure of their ventricular septal defects using the Amplatzer Septal Occluder and in the remaining three patients. Amplatzer Muscular Ventricular Septal Defect Occluder was used. The median age was 15 months (range 14 months to 10 years) and the median weight was 7.8 kg (range 7-16 kg). The devices were deployed antegradely in all patients. Immediate complete closure was obtained in three patients while two patients had small and one had a large residual following the procedure. The later was due to another multiple muscular septal defect. Transient heart block occurred in one patient during the procedure and another patient developed heart block on day 3 post-procedure. The latter required temporary pacemaker. During the follow up, one patient who had residual multiple muscular defects underwent a successful redo transcatheter closure using two Amplatzer Muscular Ventricular Septal Occluder devices. CONCLUSIONS: We conclude that transcatheter closure of muscular ventricular septal defect using Amplatzer devices is feasible and effective. PMID- 16473786 TI - Neonatal isolated critical aortic valve stenosis: balloon valvuloplasty or surgical valvotomy. AB - BACKGROUND: Open surgical valvotomy and transcatheter balloon valvuloplasty are recognised treatments for neonatal critical aortic stenosis. METHODS: A retrospective analysis was undertaken of all newborns with critical aortic valve stenosis between 1990 and 2000 presenting to a tertiary centre and who required intervention. The initial catheter and surgical intervention was generally based on the preference of the attending cardiologist and the anatomy of the aortic valve and in consultation with the cardiothoracic surgeon. The two groups were therefore not strictly comparable. Twelve were subjected to balloon valvuloplasty and thirteen to surgical valvotomy at a median age of 11 days (2-42 days) and 3.5 days (1-19 days) respectively. There was no significant difference in the timing of the procedure, weight of the infant, aortic annulus or left ventricular dimensions in either group. RESULTS: There was one unrelated hospital death in the balloon group compared to two in the surgical group both of whom had endocardial fibroelastosis. Mild to moderate aortic regurgitation was seen after both procedures. Four patients in the balloon valvuloplasty group, developed femoral artery thrombosis and two had cardiac perforation that resolved with non operative management. The mean Doppler gradient was reduced from 44+/-14 mmHg to 13.4+/-5 mmHg (p<0.01) in the valvuloplasty group compared to a reduction from 42+/-15 mmHg to 27+/-8 mmHg (p<0.05) in the surgical group. Five patients in the balloon group required re-intervention within 3 weeks to 21 months after the initial procedure. Two patients in the surgical group required a pulmonary autograft and Konno Procedure 3 and 5 years following surgical valvotomy. CONCLUSION: Both aortic valvuloplasty and valvotomy offered effective short and medium term palliation. Balloon valvuloplasty patients had a higher re intervention rate but shorter hospital and intensive care stay, reduced immediate morbidity and were associated with less severe aortic regurgitation. PMID- 16473787 TI - Pulmonary parasitosis: applied clinical and therapeutic issues. AB - BACKGROUND: The aim of this study is to review the problems encountered in treating pulmonary hydatid and bilharsiasis and to highlight the risks associated with chemotherapy and the delay of their surgical treatment. METHODS: The medical records of 60 patients with pulmonary hydatid and bilharsiasis were retrospectively investigated. The patients were divided into two groups based on whether the parasite was hydatid (group 1, n=56) or schistosomiasis (group 2, n=4). The group 1 was divided into group 1a (complicated cyst n=32) and group 1b (n=24 noncomplicated hydatids). RESULTS: In all cases of pulmonary bilharsiasis and intact pulmonary hydatid cysts, the lesions were either incidental findings or the patient had presented with haemoptysis, cough, dyspnea and chest pain. The differences between the groups with respect to the rates of preoperative complications and postoperative morbidity, frequency of decortication and hospital stay were statistically insignificant (p>0.05). CONCLUSIONS: Complicated cases have higher rates of preoperative and postoperative complications but the differences are insignificant. Complicated cases are easier to diagnose. This underlines the need for paying more attention to the possibility of pulmonary parasitosis especially in lower lobe lesions in endemic areas. PMID- 16473788 TI - Atrial distribution of connexin 40 and 43 in patients with intermittent, persistent, and postoperative atrial fibrillation. AB - BACKGROUND: Sustained atrial fibrillation (AF) causes alterations in atrial electrical and structural properties. Conflicting data regarding the structural remodeling of the gap junction proteins connexin (Cx) 40 and 43 in human and animal studies exists. We investigated the amount and distribution of Cx40 and Cx43 in three subtypes of AF. METHODS: In 50 patients undergoing coronary artery bypass graft and/or mitral or aortic valve surgery, right atrial appendages were taken and examined with immunoconfocal microscopy. Retrospectively, four groups were built: (1) sinus rhythm pre- and postoperative (SR, n=20), (2) intermittent AF, but SR prior to surgery (intAF, n=6), (3) postoperative AF (popAF, n=12), and (4) persistent AF, at least 3 month prior to surgery (persAF, n=12). We analyzed the amount of Cx40 and Cx43 and the degree of fibrosis in three randomly selected areas of each sample. RESULTS: As compared with SR, the amount of Cx40 was significantly reduced by 53% in persAF. The distribution pattern of Cx40 was heterogeneous in patients with SR, intAF, and popAF, whereas patients with persAF showed similar densities of Cx40 in the three examined areas. We found no significant difference in the amount of Cx43 between the four groups. The distribution pattern of Cx43 was heterogeneous in all four groups. The Cx40/Cx43 ratio was significantly reduced in patients with popAF and persAF by 51% and 53%, respectively. No difference was seen in the degree of fibrosis between the four groups. CONCLUSIONS: In this study, sustained AF leads to a reduction in the amount of Cx40. Together with a specific Cx40/Cx43 ratio, this may contribute to localized conduction abnormalities, facilitating the self-perpetuation of re entry pathways in AF. In the time course of structural atrial remodeling these changes seem to be earlier than a concomitantly developing fibrosis. PMID- 16473789 TI - Right atrial trans-septal approach for left atrial myxomas--nine-year experience. AB - OBJECTIVE: In this single-center study we reviewed our experience with left atrial myxomas occurring over the past 9 years. METHODS: Sixty-three patients underwent excision of cardiac myxomas between 1995 and 2004. Of these, 56 patients (32 females and 24 males) had left atrial (LA) myxoma. The mean age was 37.80+12.97 years (range 3.5-67 years). Echocardiography was the only diagnostic evaluation done. The preferred approach for resection was right atrial trans septal. Annual echocardiographic evaluation was undertaken following surgery. Follow-up is current and available in all the survivors (range 4 months-9 years). RESULTS: Clinically 75% of the LA myxomas simulated mitral stenosis. The symptoms were present for 2-6 months before operation. Most (86%) LA myxomas were attached to the fossa ovalis. Few (14%) originated from the LA wall, mitral valve annulus and anterior mitral leaflet. One patient succumbed to low cardiac output and another died of massive embolic stroke following surgery. There were no late deaths. Two patients (3.7%) developed left hemiparesis after operation but recovered completely. There was one (1.9%) recurrence 3 years after surgery. Atrial fibrillation occurred in one patient. Mitral insufficiency which was seen in two (3.7%) patients prior to surgery subsided following excision of the tumor. Postoperatively 94% patients remained without symptoms. CONCLUSION: Owing to the risk of valvular obstruction or embolization early surgery is indicated. Right atrial trans-septal approach is safe and easy. Most patients are asymptomatic following surgery. A yearly follow-up is essential. PMID- 16473790 TI - The foundation and launch of the Melbourne Interventional Group: a collaborative interventional cardiology project. AB - The Melbourne Interventional Group (MIG) is a voluntary collaborative venture of interventional cardiologists practicing at 12 major public and private hospitals in Victoria, designed to record data pertaining to percutaneous coronary interventions (PCI) and perform long-term follow-up. The potential advantages of collaboration involve large-scale analysis of current interventional strategies (e.g. drug-eluting stents, evaluation of new technologies and cost-effective analysis), provide a basis for multi-centred clinical trials and allow comparison of clinical outcomes with cardiac surgery. The established registry documents demographic, clinical and procedural characteristics of consecutive patients undergoing PCI and permits analysis of those characteristics at 30 days and 12 months. The registry is co-ordinated by the Centre of Clinical Research Excellence (CCRE), a research body within the Department of Epidemiology and Preventive Medicine (Monash University, Melbourne). The eventual goal of MIG is to provide a contemporary appraisal of Australian interventional cardiology practice, with opportunities to improve in-hospital and long-term outcomes of patients with coronary artery disease. PMID- 16473791 TI - Mitral valve replacement with a bioprosthesis: prevention of suture entanglement. AB - A simple technique of prevention of suture entanglement in the struts of bioprosthesis during implantation in the mitral position is described. PMID- 16473792 TI - Thoracic spinal cord ischemia following acute myocardial infarction and cardiac arrest in a young male. AB - In an era of percutaneous stenting for acute myocardial infarction (AMI), a case of thoracic spinal cord ischemia following AMI and cardiac arrest is presented, to highlight and discuss this rare but debilitating condition, well-documented within the neurological literature, but rarely encountered in cardiovascular practice. PMID- 16473793 TI - Pre-operative use of levosimendan in two patients with severe aortic stenosis and left ventricular dysfunction. AB - We describe two patients with severe aortic stenosis, coronary artery disease, severe left ventricular dysfunction and heart failure in which the calcium sensitising agent, levosimendan was administered prior to aortic valve replacement and coronary artery bypass graft surgery. In both cases, drug infusion was well tolerated at the doses used, heart failure improved significantly prior to surgery and peri-operative management was relatively uncomplicated in cases that would traditionally be considered high risk. Further investigation of the use of levosimendan both for treating heart failure in the presence of severe aortic stenosis and as pre-operative therapy is warranted. PMID- 16473794 TI - Subacute anthracycline cardiotoxicity. AB - We describe the case of 25-year-old man, with acute myeloid leukaemia, who presented with a myopericarditis syndrome 17 days post consolidation chemotherapy with high dose cytarabine and idarubicin. Transthoracic echocardiography showed marked transient increased left ventricular wall thickness associated with normal systolic contraction. In conjunction, pulsed tissue Doppler analysis revealed low early diastolic annular velocities, consistent with diastolic dysfunction. Endomyocardial biopsy showed severe interstitial myocardial oedema in the absence of a cellular infiltrate or myofibre damage. We believe this is the first case of subacute anthracycline toxicity described with the pathological findings of isolated myocardial oedema. PMID- 16473795 TI - Undifferentiated sarcoma of the pulmonary artery mimicking pulmonary thromboembolic disease. AB - Primary pulmonary artery sarcoma is an uncommon tumor. Mandelstamm in 1923 was the first to describe the disease in an autopsy. Since then, less than 200 cases were reported. The incidence is 0.001-0.03%, they are always highly malignant sarcoma, and women are involved twice as often as men. The presentation is often cough, dyspnea, and chest pain, and patients are usually diagnosed as suffering from pulmonary emboli, and primary tumor of the pulmonary artery is not usually considered in the differential diagnosis. The diagnosis of pulmonary artery sarcoma is made of the "clot" resected during pulmonary artery thrombendarterectomy. Our suggestion is that in patients with unilateral pulmonary artery occlusive disease, no evidence of positive hypercoagulability tests, and no history of thromboembolism, a high suspicion of pulmonary artery sarcoma should be kept in mind, and an angiographic-guided biopsy from the intra arterial occlusive material should be considered. The treatment is surgery. The survival without operation is less than 2 months. Some patients were treated with adjuvant chemotherapy. We report on a woman with undifferentiated sarcoma of the pulmonary artery, mimicking chronic pulmonary artery emboli. This case illustrates the need to consider malignancy in the differential diagnosis of patients having pulmonary emboli. PMID- 16473796 TI - A rare and life threatening complication of prosthetic valve endocarditis. AB - Up to 70% of cases of Guillain-Barre syndrome (GBS) follow a preceding infection and a number of infectious agents have been described. We present a previously unreported association of Streptococcus viridans infective endocarditis affecting a prosthetic aortic valve and Guillain-Barre syndrome. This case highlights that this potentially life-threatening diagnosis should be considered in any patient presenting with symptoms of peripheral nervous system dysfunction following an infective illness. PMID- 16473797 TI - Preface. PMID- 16473798 TI - Gastrointestinal malignancies evaluated with (18)F-fluoro-2-deoxyglucose positron emission tomography. AB - (18)F-fluoro-2-deoxyglucose positron emission tomography has demonstrated high accuracy in the staging and evaluation of colorectal and esophageal carcinomas. FDG PET is demonstrating increasing utility in a number of other gastrointestinal tumours and clinical scenarios. The established clinical indications for its use, the diagnostic accuracy, and limitations will be reviewed. Data on the emerging indications and limitations for pancreatic, hepatocellular, and gastric carcinomas, as well as gastrointestinal stromal tumours, cholangiocarcinoma, and carcinoma of unknown primary will also be briefly discussed. The use of combined PET-CT is demonstrating further improvements in diagnostic accuracy. PMID- 16473799 TI - Capsule endoscopy in 2005: facts and perspectives. AB - Capsule endoscopy has recently been introduced to explore endoscopically the whole small intestine, fulfilling a gap between examinations of the upper and lower gastrointestinal tract. The technique consists of a miniaturized endoscope, embedded in a swallowable capsule that is propulsed by peristalsis and achieves the journey to the right colon in five to eight hours. Images captured by the capsule are recorded on a hard drive worn in a belt by the patient. The main indication for capsule examination is the examination of the small bowel to find a bleeding lesion in patients with obscure bleeding. Several studies have shown that the diagnostic yield of capsule endoscopy is superior to that of push enteroscopy in this indication. Other possible indications are patients with suspected intestinal location of Crohn's disease, familial adenomatous polyposis, complicated coeliac disease and lesions due NSAIDs. The review contains information on the technical aspects of capsule endoscopy and discusses the indications. Issues of safety and tolerance are also discussed. PMID- 16473800 TI - Spectroscopy and fluorescence in esophageal diseases. AB - Malignant tumors of the esophagus continue to be a major health issue associated with high mortality primarily because most present with symptoms of dysphagia or anaemia. The disease at that stage is advanced and not likely curable. The big issue for squamous dysplasia and that associated with BE is that only a small proportion are discovered in surveillance programs when they are asymptomatic, either because the patient lives in a high-incidence geographical area, has a family history, previously diagnosed head and neck cancer or chronic reflux, as in Barrett's. Current endoscopic methods are hampered by the endoscopist's inability to recognize subtle topographic clues of dysplasia, sampling errors related to biopsy protocols, and confounding inflammation-induced artifacts both for the endoscopist and pathologist. What is desperately needed would be a biomarker (e.g. serological, fecal, urinary) that selects patients for endoscopy. However, such a test is not yet on the horizon. This article examines the current status in practice and research of novel optically based 'bioendoscopic' devices (i.e. fluorescence spectroscopy and imaging, confocal fluorescence microendoscopy (CFM), light scattering spectroscopy (LSS), Raman spectroscopy (RS), and immunophotodiagnostic endoscopy) which may enhance the diagnosis of dysplasia in all patients undergoing conventional white light endoscopy. Perhaps these new technologies will lead to more cost-effective diagnosis, mapping (e.g. surface), and staging (e.g. depth) of dysplasia, thereby allowing timely cure by endoscopic means (e.g. EMR and/or PDT), biological interventions (e.g. Cox-2 inhibitors) rather than esophajectomy. PMID- 16473801 TI - Magnifying chromoendoscopy for the detection of premalignant gastrointestinal lesions. AB - The prognosis for patients with malignancies of the gastrointestinal tract is strictly dependent on the early detection of premalignant and malignant lesions. At present, endoscopy can be performed with new, powerful high-resolution or magnifying endoscopes. Comparable to the rapid development in chip technology, the optic features of the newly designed endoscopes offer resolutions that allow new mucosal surface details to be seen. In conjunction with chromoendoscopy, the newly discovered tool of video endoscopy is much easier to use and more impressive than previously used fibreoptic endoscopy. This review summarises the value of magnifying endoscopy in the upper and lower gastrointestinal tract and focuses on gastroesophageal reflux disease and early gastric and colorectal cancer. PMID- 16473802 TI - Virtual colonoscopy in the detection of colonic polyps and neoplasms. AB - First introduced in 1994, CT colonography (Virtual colonoscopy) has emerged as an accurate, non-invasive test that will likely play a future role in colorectal cancer screening. Over the past 3 years, there have been dramatic improvements in both hardware and software technology relating to CT colonography resulting in shorter scan times, enhanced user-friendliness and improved performance statistics. Published results show the accuracy of CT colonography to be comparable to conventional colonoscopy for detection of polyps >6mm in size with few false-positives. While many of the technical aspects of CT colonography have now been standardised current interest focuses on the development of faecal tagging agents to avoid full bowel catharsis and the use of low dose multislice CT acquisition to reduce patient radiation exposure. This chapter will summarise the development of CT colonography to date, document its published performance in detection of colorectal polyps and cancers, and review its current and potential future uses. PMID- 16473803 TI - Bowel ultrasound in Crohn's disease. AB - Most published studies have found bowel ultrasound to be a useful tool in the management of Crohn's disease. Indeed, it has been successfully used as the imaging technique of choice in screening patients with clinically suspected Crohn's disease. In these patients, bowel ultrasound, which is well accepted by patients, non-invasive and of low cost, may be the first diagnostic tool employed for young patients and can be used in the preliminary diagnostic work-up prior to further invasive tests. The most important application of bowel ultrasound is, however, in the follow-up of patients already diagnosed with Crohn's disease, in whom it may be useful to assess the site and extent of the lesions and to ensure the early detection of intra-abdominal complications, particularly abscesses and strictures. In this regard, improving the ultrasound assessment of intramural blood flow by means of colour power-Doppler ultrasonography and intravenous contrast agents may help to differentiate fibrotic and inflammatory strictures, and to discriminate inflammatory masses from intra-abdominal abscesses. Despite several attempts to correlate ultrasound findings with clinical and biochemical activity, there are as yet no convincing data on the usefulness of ultrasound in assessing the activity of Crohn's disease. In contrast, preliminary results are in agreement regarding the usefulness of ultrasound in the assessment of postoperative recurrence and in monitoring the outcome of the disease following surgery. In fact, the persistence of a thickened bowel wall or increased high bowel wall thickening at ultrasound following surgery has been identified as an index of early surgical recurrence. How these data may be usefully employed in the management of individuals with Crohn's disease needs to be investigated in further studies. PMID- 16473804 TI - Anorectal ultrasound for neoplastic and inflammatory lesions. AB - Accurate staging of rectal and anal carcinoma is crucial for planning surgery and indicating adjuvant therapy. Although, computed tomography and magnetic resonance imaging are very sensitive in detecting metastatic disease, the local staging of rectal cancer with these techniques has been disappointing. Endorectal ultrasound (ERUS) and anal endosonography (AE) remain the most accurate methods for staging rectal and anal cancer. Anal endosonography is also of value in evaluating perianal sepsis: it can assist the surgeon in planning the surgical strategy by delineating the anatomy of fistula tracts, and can aid in puncturing abscesses in the operating room. Continued research and development has made the instrumentation for ERUS and AE more accurate and user-friendly. New techniques that have contributed significantly to the evolution of ERUS include three dimensional ERUS, high-frequency miniprobes, transrectal ultrasound-guided biopsy techniques and hydrogen peroxide-enhanced endosonography. Further improvements can be expected from contrast enhancement with microbubbles and colour Doppler imaging. In this new millennium, new developments in ERUS and anal endosonography, such as tri-dimensional ERUS and anal endosonography and radial electronic probing, widen the role of ERUS in the staging of rectal and anal carcinoma, as well as for perianal inflammatory conditions. PMID- 16473805 TI - Magnetic resonance imaging evaluation of small intestinal Crohn's disease. AB - Imaging evaluation in patients with Crohn's disease is based on morphological abnormalities, traditionally disclosed by barium studies including conventional enteroclysis, and more recently by cross-sectional imaging. A major prerequisite for optimal image quality and a more confident diagnosis is the adequate luminal distension of the bowel, which is usually achieved by administering an intraluminal contrast agent through a nasojejunal catheter. Magnetic resonance enteroclysis is a new technique that combines the advantages of volume challenge with state-of-the-art ultrafast pulse sequences, resulting in an excellent demonstration of the anatomy of the small bowel. Magnetic resonance enteroclysis compares favourably with conventional enteroclysis in detecting, localising and assessing the extent of involved small bowel segments. Luminal narrowing and extramural manifestations or complications of the disease can also be accurately assessed by the technique. Imaging features including the presence of deep ulcers, extensive wall thickening and mesenteric lymph nodes exhibiting marked gadolinium enhancement correlate strongly with disease activity. Although the clinical utility of magnetic resonance enteroclysis in Crohn's disease has been widely recognised, its routine application is currently limited to academic centres. Its clinical indications may include follow-up studies of known disease, the classification of Crohn's disease subtypes, an estimation of disease activity and a determination of the extramucosal extent and spread of the disease process. PMID- 16473806 TI - Magnetic resonance imaging in the detection of pancreatitis and pancreatic neoplasms. AB - This article presents current magnetic resonance imaging techniques for the diagnosis of acute and chronic pancreatitis and pancreatic neoplasms. It emphasises the need for a comprehensive protocol combining imaging sequences of the pancreatic parenchyma with magnetic resonance angiography and secretin enhanced magnetic resonance cholangiopancreatography in order to evaluate the full range of pancreatic inflammatory and neoplastic conditions. Imaging characteristics that may indicate a specific diagnosis are discussed. PMID- 16473809 TI - The physiology of cough. AB - Cough is comprised of three phases (inspiratory, compressive and expiratory) and serves as a vital defensive mechanism for lung health. It prevents pulmonary aspiration, promotes ciliary activity and clears airway debris. The importance of an intact cough mechanism is reflected in the occurrence of pulmonary problems when cough is inefficient. Cough efficiency is dependent on physical/mechanical aspects (respiratory muscles, mucus, airway calibre and larynx) and integrity of the neurophysiological pathway of cough. The understanding of the latter has progressed significantly (albeit mostly in animals) with the discovery of vanniloid receptors (and subtypes) and, more recently, by the characterisation of distinct cough receptors. However, the relative contributions of previously described airway afferents/receptors to cough are still disputed. Plasticity of the peripheral and central afferent pathways in cough has recently been shown to be important in pathological states associated with increased cough. To date, little is known of the developmental aspects of cough. PMID- 16473807 TI - Double-balloon endoscopy for the diagnosis and treatment of small intestinal disease. AB - Evaluation of the small bowel has long been hampered because of its long and multiple complex loop configurations. Yamamoto et al have developed a new method of inserting an endoscope, known as the double-balloon method of enteroscopy, the Fujinon double-balloon endoscopy system being specialised for this application. Two types of endoscope are currently available for this technique. The EN-450 P5 is a thin endoscope for regular use, whereas the EN-450 T5 is a therapeutic double-balloon endoscope with a larger accessory channel of 2.8mm in diameter. Double-balloon endoscopy enables visualisation of the entire small bowel and also allows for interventional therapy in the small intestine. This method can be used either from an oral or an anal insertion. Observation of an affected area with controlled movement of the endoscope enables interventions, including biopsies, haemostasis, balloon dilatation, stent placement, polypectomy and endoscopic mucosal resection, to be performed. In our large study series, it was demonstrated that double-balloon endoscopy permitted exploration of the small intestine with the high success rate of total enteroscopy. The procedure is safe and useful, and it provides high diagnostic yields and therapeutic capabilities. PMID- 16473810 TI - Cough in children: when does it matter? AB - Cough is a common presenting symptom to paediatricians and paediatric respiratory physicians. The causes of chronic cough in childhood are significantly different from those in adults, and a different approach to diagnosis and management is needed. In most cases cause can be identified or a satisfactory label given. This review will cover the approach to cough in children and cover some specific causes of troublesome cough not discussed elsewhere in this mini-symposium. PMID- 16473811 TI - Investigation and management of suppurative cough in pre-school children. AB - Suppurative cough can be defined as a cough where purulent sputum is produced. Chronic suppurative cough may be associated with the destruction of the bronchial wall (bronchiectasis). As mild forms of the disease are not associated with respiratory limitation or failure to thrive, such children may not present for investigation and therefore the true incidence of suppurative cough is difficult to gauge. Chronic suppurative cough remains an important health problem in developing countries and some indigenous populations of developed countries. The purpose of this review is to present the appropriate investigations and evaluate the evidence for current management strategies in children with suppurative cough. To accomplish this, a brief discussion on the aetiology of suppurative cough in childhood is presented. The most commonly identifiable cause of suppurative cough is cystic fibrosis. A detailed discussion on cystic fibrosis is beyond the scope of this review. Other causes of chronic suppurative cough in pre school children may be classified according to congenital malformations of the airway, immunodeficiency, ciliary dysfunction and, unusually, acquired causes. Microbiology of sputum culture or bronchoalveolar lavage, assessment of immune function, the role of exhaled nitric oxide and ciliary studies, and medical imaging are discussed in detail. One can conclude that the evidence for management strategies for children with suppurative cough is, at best, level 3 evidence, i.e. non-randomised, controlled or cohort studies. PMID- 16473813 TI - Cough and asthma. AB - The relationship between cough and asthma is complex. Epidemiological studies now suggest that most children with recurrent cough who do not wheeze do not have asthma. These children are commonly described as having non-specific cough that appears to be due to increased cough receptor sensitivity during the coughing episode. Children with asthma who cough have also been shown to have increased cough receptor sensitivity during an acute exacerbation. Cough severity does not generally correlate with asthma severity. Apart from wheeze and dyspnoea, there are no clear distinguishing features to separate asthma from non-specific cough. To date, no specific treatment has clearly been shown to benefit children with non-specific cough. Although a trial of asthma treatment may be justified in these children, it is preferable to cease rather than escalate treatment if there is no response. In children with asthma who cough, cough should not be used as the predominant symptom to direct asthma therapy. PMID- 16473812 TI - Habit cough: assessment and management. AB - Habit cough is a condition that is often misdiagnosed as asthma. The cough is bizarre in nature, troublesome to those around the person coughing and clearly a waking phenomenon. Often, relatives will have considered the possibility of a habit cough by the time that they present to the respiratory or general paediatrician. In the majority of cases, simple explanation of the nature of likely stressors and reassurance form the basis of effective therapy. In young people with more entrenched symptoms, the provision of coping strategies and increasing the subjective sense of control is an intervention in itself and will improve the likelihood of a good outcome. In more extreme cases, the role of rehabilitation programmes involving negotiation with schools and community organisations may prove useful in remediation of the cough and normalisation of social and peer supports. PMID- 16473814 TI - Does post-nasal drip cause cough in childhood? AB - The proposition that post-nasal drip (PND) is a common cause of cough in childhood is controversial. The concept of PND as a common cause of cough is derived primarily from the adult literature. The definition of what constitutes PND is variable and it is unclear whether it is a symptom, a sign or both. Examination of the specificity of symptoms and signs for the diagnosis of PND syndrome further confuses the issue. A definitive diagnosis of cough induced by PND cannot be made from history and physical examination alone. The concept is inconsistent both with the meaning of the word 'drip' and the science of rheology. The most plausible explanation for the occurrence of cough in children identified with increased post-nasal secretions is that both reflect co-existent airways pathologies. In considering causes of chronic cough in childhood, it is now time to abandon the concept of PND and the associated PND syndrome. PMID- 16473815 TI - Secondary pulmonary hypertension. AB - Recent research has greatly improved our understanding of the pathophysiology of pulmonary hypertension. There is increasing recognition that pulmonary hypertension is an important complication of many childhood respiratory diseases including cystic fibrosis, interstitial lung diseases, upper airways obstruction and disorders of the respiratory muscles and chest wall. Chronic hypoxaemia and, in some cases, destruction of the vascular bed are the key factors. The clinical features of pulmonary hypertension are often overshadowed by those of the primary respiratory disease but newer imaging techniques allow earlier detection of this complication. This may be important in the future if new specific therapies for primary pulmonary hypertension are shown to be beneficial in secondary pulmonary hypertension. With some conditions, such as airways obstruction due to adenotonsillar hypertrophy, treating the underlying cause leads to rapid resolution of the hypertension. However, with most disorders, such as cystic fibrosis, management is focused on treating the lung disease intensively and correcting the chronic hypoxaemia with oxygen therapy, sometimes augmented by nasal mask ventilation. However, although several new selective therapies are effective in idiopathic pulmonary arterial hypertension, their role in secondary pulmonary hypertension remains unclear. PMID- 16473816 TI - Pulmonary haemosiderosis in infants and children. AB - Pulmonary haemosiderosis (PH) results from recurrent bleeding into alveolar spaces and interstitial lung tissue. If untreated, fibrosis and restrictive lung disease will develop and may lead to death. A distinction can be made between primary and secondary pulmonary haemosiderosis as a manifestation of auto-immune small vessel vasculitides, coagulation disorders or diseases with increased venous pressure. Interestingly, in some cases of 'idiopathic' PH, exposure to moulds and van Willebrand's disease may contribute to the pathogenesis. Haemosiderosis usually begins with haematemesis. Anaemia, alveolar infiltrates on chest x ray and siderophages in broncho-alveolar lavage fluid are diagnostic factors. Immunosuppressive treatment has improved the prognosis in recent years. PMID- 16473817 TI - Transgenic mice. AB - For the past 20 years researchers have used transgenic mice to help understand the basic mechanisms associated with inherited human and animal diseases. The ability to integrate exogenous genetic information into the mouse genome has revolutionised the analysis of gene function. Both gene addition and gene replacement can be performed and the capability exists to create 'conditional' mutations and to study gene dosage effects. The aim of the present review is to provide a framework of information on transgenic mouse methodologies that can be applied to any area of research. A basic understanding of transgenic technology, recognising its advantages and disadvantages, is essential knowledge for the scientist in the 21st century. PMID- 16473818 TI - The relationship between paediatricians and commerce. AB - The interaction between doctors and commerce, particularly the pharmaceutical industry, has recently been subject to increasing scrutiny. Doctors are now exposed to mounting influence from industry as it spends large amounts of money on marketing, is heavily involved with continuing medical education and sponsors a major proportion of research. Conflicts of interest may exist on both sides of the relationship: doctors can be manipulated and companies need to be profitable. Paediatricians are just as open to this influence as are other members of the medical profession. There is evidence that clinical practice is altered by interaction with industry, although doctors appear to deny the likelihood of being influenced. There are significant concerns over the increasing involvement of the pharmaceutical industry with research, although the regulation of the industry continues to be strengthened and the process of research is becoming more transparent. Disclosure of conflicts of interest involving authorship is now common practice and should extend to all facets of the relationship. However, collaboration continues to be necessary in order to develop new therapies, maximise research and particularly in paediatrics, to test medications in children. Paediatricians need to be aware of the sources of influence and understand current guidelines so that interactions with industry continue to be appropriate. PMID- 16473819 TI - Paediatricians and the pharmaceutical industry: an industry perspective of the challenges ahead. AB - The relationship between healthcare professionals and the pharmaceutical industry is under intense scrutiny. Accusations of corrupt practices have been levelled at the industry by both the professional and the lay press. The environment is changing, with rising expectations of transparency and ethical standards. In addition, society is becoming increasingly risk averse. There have been examples of poor practice by industry in the past, and industry is learning from these. Equally, there are many examples of excellent practice where industry has worked effectively and ethically with clinicians. The goal of industry is to bring new medicines to benefit patients and shareholders. Commercial success is dependent upon putting the patient at the centre of activity. It also allows re-investment into research and development. This allies industry and healthcare professionals with a common and key ethical arbitrator--the patient. Industry is changing as an acknowledgement of the need to adapt to the culture change demanded by society. Self regulation is increasingly active and transparent; guidelines, laws and internal/external regulators exist to examine industry. However, for the changes in regulations, behaviour and practices to be fully effective, there is a need for dialogue between industry and healthcare professionals. Commitment is needed from both sides to work together to manage the relationship. PMID- 16473820 TI - Allergic bronchopulmonary aspergillosis in paediatric cystic fibrosis patients. AB - Allergic bronchopulmonary aspergillosis (ABPA) is a severe complication in children, adolescents and adults with cystic fibrosis (CF), the prevalence of which ranges from 6-25%. The disease is the result of the colonisation of the respiratory tract by fungi of the genus Aspergillus, commonly Aspergillus fumigatus, and subsequent host sensitisation to fungal antigens, accompanied by a Th2 CD4 type response mediated by the production of specific IgE. The consequent inflammatory and obstructive bronchopulmonary injury can progress to fibrosis. The diagnosis should be considered early in patients with CF who show wheezing, transient pulmonary infiltrates and reduced lung function. The objective diagnosis is not straightforward because of overlapping clinical and radiological signs, particularly the progression of bronchiectasis. Specific criteria are needed for the diagnosis of ABPA in patients with CF, such as those proposed by the Cystic Fibrosis Foundation. The study of specific IgE against recombinant antigens of A. fumigatus has contributed to the early diagnosis of ABPA with high sensitivity and specificity. The technique has also shown promise in the follow up of patients after steroid therapy and the early detection of recurrences. Treatment consists of long-term systemic corticosteroid usage, the monitoring of their adverse effects, and of the measurement of total serum IgE levels. The concomitant use of oral itraconazole seems to promote a better control of the disease and to reduce the duration of systemic steroid therapy but its use continues to be controversial. Controlled studies involving larger numbers of patients are necessary if we are to better understand the management of ABPA. PMID- 16473821 TI - The assessment and management of croup. AB - The treatment of croup has changed considerably over the last 25 years with the liberalisation of the use of systemic corticosteroids for mild to moderate croup. The administration of corticosteroids in croup has reduced the severity of the condition, dramatically reduced the need for endotracheal intubation, shortened the duration of intubation, reduced the length of hospital stay, reduced the need for hospital admission and reduced daycare/preschool absenteeism and improved sleep in milder cases. Despite studies showing the efficacy of nebulised and intramuscular corticosteroids, the use of oral corticosteroids remains the recommended option in most, if not all, cases of croup presenting for medical assessment. PMID- 16473822 TI - What can we learn from reconstructing the extinct 1918 pandemic influenza virus? PMID- 16473823 TI - B cells discriminate the rules of engagement. AB - How B cells discriminate antigen-receptor-mediated signals in development and navigate critical checkpoints in adaptive immune responses remains poorly resolved. In this issue of Immunity, conditionally ablate the main regulatory subunit of the calcineurin phosphatase complex in B cells to reveal both positive and negative influences on the development of antibody responses and B cell memory. PMID- 16473824 TI - CD14: chaperone or matchmaker? AB - In this issue of Immunity, describe a physical and functional relationship between Toll-like receptor 3 (TLR3) and the pattern recognition protein CD14. In the presence of CD14, TLR3-mediated signal transduction events are amplified. PMID- 16473825 TI - An unexpected connection: lymph node lymphangiogenesis and dendritic cell migration. AB - Migration of antigen-presenting dendritic cells (DCs) through lymphatics to draining lymph nodes (LNs) represents a key step in the initiation of an adaptive immune response. In this issue of Immunity, provide the first evidence that remodeling of the lymphatic network within LNs and in peripheral tissues enhances DC migration after immunization and that this process depends on the presence of B cells within the LN. PMID- 16473826 TI - Deconstructing the form and function of the TCR/CD3 complex. AB - When T cells encounter antigens via the T cell antigen receptor (TCR), information about the quantity and quality of antigen engagement is relayed to the intracellular signal transduction machinery. This process is poorly understood. The TCR itself lacks a significant intracellular domain. Instead, it is associated with CD3 molecules that contain intracellular signaling domains that couple the TCR/CD3 complex to the downstream signaling machinery. The earliest events in TCR signaling must involve the transfer of information from the antigen binding TCR subunit to the CD3 signaling subunits of the TCR/CD3 complex. Elucidating the structural organization of the TCR with the associated CD3 signaling molecules is necessary for understanding the mechanism by which TCR engagement is coupled to activation. Here, we review the current state of our understanding of the structure and organization of the TCR/CD3 complex. PMID- 16473827 TI - The calcineurin phosphatase complex modulates immunogenic B cell responses. AB - A series of signal-directed transitions regulates the development of distinct populations of self-tolerant B cells and ultimately the production of antibody producing plasma cells. We studied the role of calcineurin/NFAT signaling in B cells by deleting the regulatory b1 subunit of calcineurin specifically in B cells. Follicular (FO) and marginal zone (MZ) B cells develop normally in these mice, but B1 cell numbers are reduced. In vitro, calcineurin b1-deficient B cells have a cell-intrinsic proliferation defect downstream of the B cell receptor. These mice have higher total serum IgM despite the absence of B1 cells and have enhanced T cell-independent-1 responses. Conversely, mice with calcineurin b1 deficient B cells develop larger germinal centers and have reduced plasma cell development and antigen-specific antibody production during T cell-dependent immune responses. By several different criteria, calcineurin is dispensable for B cell tolerance, indicating that this phosphatase complex modulates immunogenic, but not tolerogenic, responses in vivo. PMID- 16473828 TI - Double-stranded RNA-mediated TLR3 activation is enhanced by CD14. AB - CD14 is a well-known pattern-recognition receptor in the innate immune system. Here, we show that CD14 enhances double-stranded RNA (dsRNA)-mediated Toll-like receptor 3 (TLR3) activation. Bone marrow-derived macrophages (BMDMs) from CD14-/ mice exhibited impaired responses to polyinosine-polycytidylic acid (pIpC) and reduced production of inflammatory cytokines. CD14-/- mice injected with pIpC also showed impaired cytokine production. When tested with [32P] labeled pIpC small fragments (pIpCsf) that maintain the inflammatory activity of crude pIpC, CD14 directly bound pIpCsf and mediated cellular uptake of pIpCsf. Our data show that TLR3 is intracellular and directly interacts with CD14. Internalized pIpCsf was localized in the lysosomes via the endosomes. In unstimulated cells, neither CD14 nor TLR3 was detected in the lysosomes. However, TLR3 was localized in the lysosomes as was CD14 once the cells took up pIpC. We also observed that internalized pIpCsf colocalized with CD14 and TLR3. Consequently, CD14 mediates pIpC uptake and enhances TLR3 signaling. PMID- 16473829 TI - CCR7-dependent cortex-to-medulla migration of positively selected thymocytes is essential for establishing central tolerance. AB - Immature CD4+CD8+ thymocytes, which are generated in the thymic cortex, are induced upon positive selection to differentiate into mature T lymphocytes and relocate to the thymic medulla. It was recently shown that a chemokine signal via CCR7 is essential for the cortex-to-medulla migration of positively selected thymocytes in the thymus. However, the role of the cortex-to-medulla migration in T cell development and selection has remained unclear. The present study shows that the developmental kinetics and the thymic export of mature thymocytes were undisturbed in adult mice lacking CCR7 or its ligands (CCR7L). The inhibition of sphingosine-1-phosphate-mediated lymphocyte egress from the thymus led to the accumulation of mature thymocytes in the cortex of CCR7- or CCR7L-deficient mice, unlike the accumulation in the medulla of normal mice, thereby suggesting that mature thymocytes may be exported directly from the cortex in the absence of CCR7 signals. However, the thymocytes that were generated in the absence of CCR7 or CCR7L were potent in causing autoimmune dacryoadenitis and sialadenitis in mice and were thus incapable of establishing central tolerance to organ-specific antigens. These results indicate that CCR7-mediated cortex-to-medulla migration of thymocytes is essential for establishing central tolerance rather than for supporting the maturation or export of thymocytes. PMID- 16473830 TI - TGFbeta in the context of an inflammatory cytokine milieu supports de novo differentiation of IL-17-producing T cells. AB - We describe de novo generation of IL-17-producing T cells from naive CD4 T cells, induced in cocultures of naive CD4 T cells and naturally occurring CD4+ CD25+ T cells (Treg) in the presence of TLR3, TLR4, or TLR9 stimuli. Treg can be substituted by TGFbeta1, which, together with the proinflammatory cytokine IL-6, supports the differentiation of IL-17-producing T cells, a process that is amplified by IL-1beta and TNFalpha. We could not detect a role for IL-23 in the differentiation of IL-17-producing T cells but confirmed its importance for their survival and expansion. Transcription factors GATA-3 and T-bet, as well as its target Hlx, are absent in IL-17-producing T cells, and they do not express the negative regulator for TGFbeta signaling, Smad7. Our data indicate that, in the presence of IL-6, TGFbeta1 subverts Th1 and Th2 differentiation for the generation of IL-17-producing T cells. PMID- 16473831 TI - Dendritic cells rapidly recruited into epithelial tissues via CCR6/CCL20 are responsible for CD8+ T cell crosspriming in vivo. AB - The nature of dendritic cell(s) (DC[s]) that conditions efficient in vivo priming of CD8+ CTL after immunization via epithelial tissues remains largely unknown. Here, we show that myeloid DCs rapidly recruited by adjuvants into the buccal mucosa or skin are essential for CD8+ T cell crosspriming. Recruitment of circulating DC precursors, including Gr1+ monocytes, precedes the sequential accumulation of CD11c+ MHC class II+ DCs in dermis and epithelium via a CCR6/CCL20-dependent mechanism. Remarkably, a defect in CCR6, local neutralization of CCL20, or depletion of monocytes prevents in vivo priming of CD8+ CTL against an innocuous protein antigen administered with adjuvant. In addition, transfer of CCR6-sufficient Gr1+ monocytes restores CD8+ T cell priming in CCR6( degrees / degrees ) mice via a direct Ag presentation mechanism. Thus, newly recruited DCs likely derived from circulating monocytes are responsible for efficient crosspriming of CD8+ CTL after mucosal or skin immunization. PMID- 16473833 TI - Dynamics of thymus-colonizing cells during human development. AB - Here, we identify fetal bone marrow (BM)-derived CD34hiCD45RAhiCD7+ hematopoietic progenitors as thymus-colonizing cells. This population, virtually absent from the fetal liver (FL), emerges in the BM by development weeks 8-9, where it accumulates throughout the second trimester, to finally decline around birth. Based on phenotypic, molecular, and functional criteria, we demonstrate that CD34hiCD45RAhiCD7+ cells represent the direct precursors of the most immature CD34hiCD1a- fetal thymocytes that follow a similar dynamics pattern during fetal and early postnatal development. Histological analysis of fetal thymuses further reveals that early immigrants predominantly localize in the perivascular areas of the cortex, where they form a lymphostromal complex with thymic epithelial cells (TECs) driving their rapid specification toward the T lineage. Finally, using an ex vivo xenogeneic thymus-colonization assay, we show that BM-derived CD34hiCD45RAhiCD7+ progenitors are selectively recruited into the thymus parenchyma in the absence of exogenous cytokines, where they adopt a definitive T cell fate. PMID- 16473832 TI - B cell-driven lymphangiogenesis in inflamed lymph nodes enhances dendritic cell mobilization. AB - Dendritic cell (DC) migration from the periphery to lymph nodes is regulated by the pattern of genes expressed by DCs themselves and by signals within the surrounding peripheral environment. Here, we report that DC mobilization can also be regulated by signals initiated within the downstream lymph nodes, particularly when lymph nodes enlarge as a consequence of immunization. Lymph node B lymphocytes orchestrate expansion of the lymphatic network within the immunized lymph node. This expanded network in turn supports increased DC migration from the periphery. These results reveal unique relationships between B cells, lymphatic vessels, and migratory DCs. Knowledge that DC migration from the periphery is augmented by B cell-dependent signals reveals new potential strategies to increase DC migration during vaccination. PMID- 16473835 TI - Transport of galectin-3 between the nucleus and cytoplasm. I. Conditions and signals for nuclear import. AB - Galectin-3, a factor involved in the splicing of pre-mRNA, shuttles between the nucleus and the cytoplasm. We have engineered a vector that expresses the fusion protein containing the following: (a) green fluorescent protein as a reporter of localization, (b) bacterial maltose-binding protein to increase the size of the reporter polypeptide, and (c) galectin-3, whose sequence we wished to dissect in search of amino acid residues vital for nuclear localization. In mouse 3T3 fibroblasts transfected with this expression construct, the full-length galectin 3 (residues 1-263) fusion protein was localized predominantly in the nucleus. Mutants of this construct, containing truncations of the galectin-3 polypeptide from the amino terminus, retained nuclear localization through residue 128; thus, the amino-terminal half was dispensable for nuclear import. Mutants of the same construct, containing truncations from the carboxyl terminus, showed loss of nuclear localization. This effect was observed beginning with truncation at residue 259, and the full effect was seen with truncation at residue 253. Site directed mutagenesis of the sequence ITLT (residues 253-256) suggested that nuclear import was dependent on the IXLT type of nuclear localization sequence, first discovered in the Drosophila protein Dsh (dishevelled). In the galectin-3 polypeptide, the activity of this nuclear localization sequence is modulated by a neighboring leucine-rich nuclear export signal. PMID- 16473836 TI - Enterovirus infections. PMID- 16473834 TI - Transport of galectin-3 between the nucleus and cytoplasm. II. Identification of the signal for nuclear export. AB - Galectin-3, a factor involved in the splicing of pre-mRNA, shuttles between the nucleus and the cytoplasm. Previous studies have shown that incubation of fibroblasts with leptomycin B resulted in the accumulation of galectin-3 in the nucleus, suggesting that the export of galectin-3 from the nucleus may be mediated by the CRM1 receptor. A candidate nuclear export signal fitting the consensus sequence recognized by CRM1 can be found between residues 240 and 255 of the murine galectin-3 sequence. This sequence was engineered into the pRev(1.4) reporter system, in which candidate sequences can be tested for nuclear export activity in terms of counteracting the nuclear localization signal present in the Rev(1.4) protein. Rev(1.4)-galectin-3(240-255) exhibited nuclear export activity that was sensitive to inhibition by leptomycin B. Site-directed mutagenesis of Leu247 and Ile249 in the galectin-3 nuclear export signal decreased nuclear export activity, consistent with the notion that these two positions correspond to the critical residues identified in the nuclear export signal of the cAMP-dependent protein kinase inhibitor. The nuclear export signal activity was also analyzed in the context of a full-length galectin-3 fusion protein; galectin-3(1-263; L247A) showed more nuclear localization than wild type, implicating Leu247 as critical to the function of the nuclear export signal. These results indicate that residues 240-255 of the galectin-3 polypeptide contain a leucine-rich nuclear export signal that overlaps with the region (residues 252-258) identified as important for nuclear localization. PMID- 16473837 TI - Radiation risks of CT scans. PMID- 16473838 TI - Health and well-being of children in foster care placement. PMID- 16473839 TI - Infant botulism. PMID- 16473840 TI - Syphilis. PMID- 16473841 TI - SNP genotyping using alkali cleavage of RNA/DNA chimeras and MALDI time-of-flight mass spectrometry. AB - Single nucleotide polymorphisms (SNPs) are now widely used for many DNA analysis applications such as linkage disequilibrium mapping, pharmacogenomics and traceability. Many methods for SNP genotyping exist with diverse strategies for allele-distinction. Mass spectrometers are used most commonly in conjunction with primer extension procedures with allele-specific termination. Here we present a novel concept for allele-preparation for SNP genotyping. Primer extension is carried out with an extension primer positioned immediately upstream of the SNP that is to be genotyped, a complete set of four ribonucleotides and a ribonucleotide incorporating DNA polymerase. The allele-extension products are then treated with alkali, which results in the cleavage immediately after the first added ribonucleotide. In addition, to obtain fragments easily detectable by mass spectrometry, we have included a ribonucleotide in the primer usually at the fourth nucleotide from the 3' terminus. The method was tested on four SNPs each with a different combination of nucleotides. The advantage over other mass spectrometry-based SNP genotyping assays is that this one only requires a PCR, a primer extension reaction with a universal extension mix and an inexpensive facile cleavage reaction, which makes it overall very cost effective and easy in handling. PMID- 16473843 TI - Expression of Cre recombinase during transient phage infection permits efficient marker removal in Streptomyces. AB - We report a system for the efficient removal of a marker flanked by two loxP sites in Streptomyces coelicolor, using a derivative of the temperate phage phiC31 that expresses Cre recombinase during a transient infection. As the test case for this recombinant phage (called Cre-phage), we present the construction of an in-frame deletion of a gene, pglW, required for phage growth limitation or Pgl in S.coelicolor. Cre-phage was also used for marker deletion in other strains of S.coelicolor. PMID- 16473842 TI - Combination of methylated-DNA precipitation and methylation-sensitive restriction enzymes (COMPARE-MS) for the rapid, sensitive and quantitative detection of DNA methylation. AB - Hypermethylation of CpG island (CGI) sequences is a nearly universal somatic genome alteration in cancer. Rapid and sensitive detection of DNA hypermethylation would aid in cancer diagnosis and risk stratification. We present a novel technique, called COMPARE-MS, that can rapidly and quantitatively detect CGI hypermethylation with high sensitivity and specificity in hundreds of samples simultaneously. To quantitate CGI hypermethylation, COMPARE-MS uses real time PCR of DNA that was first digested by methylation-sensitive restriction enzymes and then precipitated by methyl-binding domain polypeptides immobilized on a magnetic solid matrix. We show that COMPARE-MS could detect five genome equivalents of methylated CGIs in a 1000- to 10,000-fold excess of unmethylated DNA. COMPARE-MS was used to rapidly quantitate hypermethylation at multiple CGIs in >155 prostate tissues, including benign and malignant prostate specimens, and prostate cell lines. This analysis showed that GSTP1, MDR1 and PTGS2 CGI hypermethylation as determined by COMPARE-MS could differentiate between malignant and benign prostate with sensitivities >95% and specificities approaching 100%. This novel technology could significantly improve our ability to detect CGI hypermethylation. PMID- 16473844 TI - Isolation of a site-specifically modified RNA from an unmodified transcript. AB - Natural RNAs contain many base modifications that have specific biological functions. The ability to functionally dissect individual modifications is facilitated by the identification and cloning of enzymes responsible for these modifications, but is hindered by the difficulty of isolating site-specifically modified RNAs away from unmodified transcripts. Using the m1G37 and m1A58 methyl modifications of tRNA as two examples, we demonstrate that non-pairing base modifications protect RNAs against the DNA-directed RNase H cleavage. This provide a new approach to obtain homogeneous RNAs with site-specific base modifications that are suitable for biochemical and functional studies. PMID- 16473845 TI - BTA, a novel reagent for DNA attachment on glass and efficient generation of solid-phase amplified DNA colonies. AB - The tricarboxylate reagent benzene-1,3,5-triacetic acid (BTA) was used to attach 5'-aminated DNA primers and templates on an aminosilanized glass surface for subsequent generation of DNA colonies by in situ solid-phase amplification. We have characterized the derivatized surfaces for the chemical attachment of oligonucleotides and evaluate the properties relevant for the amplification process: surface density, thermal stability towards thermocycling, functionalization reproducibility and storage stability. The derivatization process, first developed for glass slides, was then adapted to microfabricated glass channels containing integrated fluidic connections. This implementation resulted in an important reduction of reaction times, consumption of reagents and process automation. Innovative analytical methods for the characterization of attached DNA were developed for assessing the surface immobilized DNA content after amplification. The results obtained showed that the BTA chemistry is compatible and suitable for forming highly dense arrays of DNA colonies with optimal surface coverage of about 10 million colonies/cm(2) from the amplification of initial single-template DNA molecules immobilized. We also demonstrate that the dsDNA colonies generated can be quantitatively processed in situ by restriction enzymes digestion. DNA colonies generated using the BTA reagent can be used for further sequence analysis in an unprecedented parallel fashion for low-cost genomic studies. PMID- 16473846 TI - A translation control reporter system (TCRS) for the analysis of translationally controlled processes in the vertebrate cell. AB - Regulation of translation is critical for the accurate expression of a broad variety of genes that function in cell cycle progression and cell differentiation, as well as in the adaptation to cellular stress. The aetiologies of a number of human diseases, including cancer, have been linked to mutations in genes that control mRNA translation, or in cis-regulatory mRNA-sequences. Therefore, research on translational control and its therapeutic appliance has become most important. However, to date only a limited number of therapeutic drugs are known to affect translational control. Here we describe a novel, straightforward approach for the detection of cellular translational activity. We developed a Translational Control Reporter System (TCRS), which utilizes the cis regulatory upstream open reading frame (uORF) from the c/ebpalpha locus to direct the translation of a dual reporter gene into two unique reporter peptides. The peptides contain a pre-pro-trypsin (PPT) signal for secretion into the medium and distinct immunogenic epitopes for detection and quantification purposes. TCRS peptide expression levels reflect changes of translation initiation induced by serum growth factors, drugs or translation factor mutants. TCRS can be tailored to various research settings and the system may accomplish a broad application to uncover links between translational control and drugs. PMID- 16473848 TI - Visualizing bacterial tRNA identity determinants and antideterminants using function logos and inverse function logos. AB - Sequence logos are stacked bar graphs that generalize the notion of consensus sequence. They employ entropy statistics very effectively to display variation in a structural alignment of sequences of a common function, while emphasizing its over-represented features. Yet sequence logos cannot display features that distinguish functional subclasses within a structurally related superfamily nor do they display under-represented features. We introduce two extensions to address these needs: function logos and inverse logos. Function logos display subfunctions that are over-represented among sequences carrying a specific feature. Inverse logos generalize both sequence logos and function logos by displaying under-represented, rather than over-represented, features or functions in structural alignments. To make inverse logos, a compositional inverse is applied to the feature or function frequency distributions before logo construction, where a compositional inverse is a mathematical transform that makes common features or functions rare and vice versa. We applied these methods to a database of structurally aligned bacterial tDNAs to create highly condensed, birds-eye views of potentially all so-called identity determinants and antideterminants that confer specific amino acid charging or initiator function on tRNAs in bacteria. We recovered both known and a few potentially novel identity elements. Function logos and inverse logos are useful tools for exploratory bioinformatic analysis of structure-function relationships in sequence families and superfamilies. PMID- 16473847 TI - TFAM detects co-evolution of tRNA identity rules with lateral transfer of histidyl-tRNA synthetase. AB - We present TFAM, an automated, statistical method to classify the identity of tRNAs. TFAM, currently optimized for bacteria, classifies initiator tRNAs and predicts the charging identity of both typical and atypical tRNAs such as suppressors with high confidence. We show statistical evidence for extensive variation in tRNA identity determinants among bacterial genomes due to variation in overall tDNA base content. With TFAM we have detected the first case of eukaryotic-like tRNA identity rules in bacteria. An alpha-proteobacterial clade encompassing Rhizobiales, Caulobacter crescentus and Silicibacter pomeroyi, unlike a sister clade containing the Rickettsiales, Zymomonas mobilis and Gluconobacter oxydans, uses the eukaryotic identity element A73 instead of the highly conserved prokaryotic element C73. We confirm divergence of bacterial histidylation rules by demonstrating perfect covariation of alpha-proteobacterial tRNA(His) acceptor stems and residues in the motif IIb tRNA-binding pocket of their histidyl-tRNA synthetases (HisRS). Phylogenomic analysis supports lateral transfer of a eukaryotic-like HisRS into the alpha-proteobacteria followed by in situ adaptation of the bacterial tDNA(His) and identity rule divergence. Our results demonstrate that TFAM is an effective tool for the bioinformatics, comparative genomics and evolutionary study of tRNA identity. PMID- 16473849 TI - Mechanisms of transcriptional repression of cell-cycle G2/M promoters by p63. AB - p63 is a developmentally regulated transcription factor related to p53, which activates and represses specific genes. The human AEC (Ankyloblepharon-Ectodermal dysplasia-Clefting) and EEC (Ectrodactyly-Ectodermal dysplasia-Cleft lip/palate) syndromes are caused by missense mutations of p63, within the DNA-binding domain (EEC) or in the C-terminal sterile alpha motif domain (AEC). We show here that p63 represses transcription of cell-cycle G(2)/M genes by binding to multiple CCAAT core promoters in immortalized and primary keratinocytes. The CCAAT activator NF-Y and DeltaNp63alpha are associated in vivo and a conserved alpha helix of the NF-YC histone fold is required. p63 AEC mutants, but not an EEC mutant, are incapable to bind NF-Y. DeltaNp63alpha, but not the AEC mutants repress CCAAT-dependent transcription of G(2)/M genes. Chromatin immunoprecipitation recruitment assays establish that the AEC mutants are not recruited to G(2)/M promoters, while normally present on 14-3-3sigma, which contains a sequence-specific binding site. Surprisingly, the EEC C306R mutant activates transcription. Upon keratinocytes differentiation, NF-Y and p63 remain bound to G(2)/M promoters, while HDACs are recruited, histones deacetylated, Pol II displaced and transcription repressed. Our data indicate that NF-Y is a molecular target of p63 and that inhibition of growth activating genes upon differentiation is compromised by AEC missense mutations. PMID- 16473850 TI - DNA nicking by HinP1I endonuclease: bending, base flipping and minor groove expansion. AB - HinP1I recognizes and cleaves the palindromic tetranucleotide sequence G downward arrowCGC in DNA. We report three structures of HinP1I-DNA complexes: in the presence of Ca(2+) (pre-reactive complex), in the absence of metal ion (binary complex) and in the presence of Mg(2+) (post-reactive complex). HinP1I forms a back-to-back dimer with two active sites and two DNA duplexes bound on the outer surfaces of the dimer facing away from each other. The 10 bp DNA duplexes undergo protein-induced distortions exhibiting features of A-, B- and Z-conformations: bending on one side (by intercalation of a phenylalanine side chain into the major groove), base flipping on the other side of the recognition site (by expanding the step rise distance of the local base pair to Z-form) and a local A form conformation between the two central C:G base pairs of the recognition site (by binding of the N-terminal helix in the minor groove). In the pre- and post reactive complexes, two metals (Ca(2+) or Mg(2+)) are found in the active site. The enzyme appears to cleave DNA sequentially, hydrolyzing first one DNA strand, as seen in the post-reactive complex in the crystalline state, and then the other, as supported by the observation that, in solution, a nicked DNA intermediate accumulates before linearization. PMID- 16473851 TI - Roles for Gcn5p and Ada2p in transcription and nucleotide excision repair at the Saccharomyces cerevisiae MET16 gene. AB - Chromatin structure, transcription and repair of cyclobutane pyrimidine dimers at the MET16 gene of wild type, gcn5Delta and ada2Delta Saccharomyces cerevisiae cells were studied under repressing or derepressing conditions. These two components of the SAGA/ADA chromatin remodelling complexes are expendable for the basal transcription of MET16 but are mandatory for its full transcription induction. Despite their influence on transcription neither protein induces major changes in MET16 chromatin structure, but some minor ones occur. Repair at the coding region of the transcribed strand is faster than repair at non-transcribed regions in all strains and either growth condition. Moreover, the more MET16 is transcribed the faster the repair. The data show that by changing the transcription extent the rate of repair at each DNA strand is altered in a different way, confirming that repair at this locus is strongly modulated by its chromatin structure and transcription level. Deletion of GCN5 or ADA2 reduces repair at MET16. The results are discussed in light of the current understanding of Gcn5p and Ada2p functions, and they are the first to report a role for Ada2p in the nucleotide excision repair of the regulatory and transcribed regions of a gene. PMID- 16473852 TI - Options available for profiling small samples: a review of sample amplification technology when combined with microarray profiling. AB - The possibility of performing microarray analysis on limited material has been demonstrated in a number of publications. In this review we approach the technical aspects of mRNA amplification and several important implicit consequences, for both linear and exponential procedures. Amplification efficiencies clearly allow profiling of extremely small samples. The conservation of transcript abundance is the most important issue regarding the use of sample amplification in combination with microarray analysis, and this aspect has generally been found to be acceptable, although demonstrated to decrease in highly diluted samples. The fact that variability and discrepancies in microarray profiles increase with minute sample sizes has been clearly documented, but for many studies this does appear to have affected the biological conclusions. We suggest that this is due to the data analysis approach applied, and the consequence is the chance of presenting misleading results. We discuss the issue of amplification sensitivity limits in the light of reports on fidelity, published data from reviewed articles and data analysis approaches. These are important considerations to be reflected in the design of future studies and when evaluating biological conclusions from published microarray studies based on extremely low input RNA quantities. PMID- 16473853 TI - tRNA properties help shape codon pair preferences in open reading frames. AB - Translation elongation is an accurate and rapid process, dependent upon efficient juxtaposition of tRNAs in the ribosomal A- and P-sites. Here, we sought evidence of A- and P-site tRNA interaction by examining bias in codon pair choice within open reading frames from a range of genomes. Three distinct and marked effects were revealed once codon and dipeptide biases had been subtracted. First, in the majority of genomes, codon pair preference is primarily determined by a tetranucleotide combination of the third nucleotide of the P-site codon, and all 3 nt of the A-site codon. Second, pairs of rare codons are generally under-used in eukaryotes, but over-used in prokaryotes. Third, the analysis revealed a highly significant effect of tRNA-mediated selection on codon pairing in unicellular eukaryotes, Bacillus subtilis, and the gamma proteobacteria. This was evident because in these organisms, synonymous codons decoded in the A-site by the same tRNA exhibit significantly similar P-site pairing preferences. Codon pair preference is thus influenced by the identity of A-site tRNAs, in combination with the P-site codon third nucleotide. Multivariate analysis identified conserved nucleotide positions within A-site tRNA sequences that modulate codon pair preferences. Structural features that regulate tRNA geometry within the ribosome may govern genomic codon pair patterns, driving enhanced translational fidelity and/or rate. PMID- 16473855 TI - Computational inference and experimental validation of the nitrogen assimilation regulatory network in cyanobacterium Synechococcus sp. WH 8102. AB - Deciphering the regulatory networks encoded in the genome of an organism represents one of the most interesting and challenging tasks in the post-genome sequencing era. As an example of this problem, we have predicted a detailed model for the nitrogen assimilation network in cyanobacterium Synechococcus sp. WH 8102 (WH8102) using a computational protocol based on comparative genomics analysis and mining experimental data from related organisms that are relatively well studied. This computational model is in excellent agreement with the microarray gene expression data collected under ammonium-rich versus nitrate-rich growth conditions, suggesting that our computational protocol is capable of predicting biological pathways/networks with high accuracy. We then refined the computational model using the microarray data, and proposed a new model for the nitrogen assimilation network in WH8102. An intriguing discovery from this study is that nitrogen assimilation affects the expression of many genes involved in photosynthesis, suggesting a tight coordination between nitrogen assimilation and photosynthesis processes. Moreover, for some of these genes, this coordination is probably mediated by NtcA through the canonical NtcA promoters in their regulatory regions. PMID- 16473856 TI - Epidermolysis bullosa. I. Molecular genetics of the junctional and hemidesmosomal variants. AB - INTRODUCTION: Epidermolysis bullosa (EB), a group of autosomal heritable blistering diseases, is characterised by extensive phenotypic variability with considerable morbidity and mortality. EB is classified into distinct subtypes depending on the location of blistering within the cutaneous dermoepidermal basement membrane zone. Ten genes are known to harbour mutations in the major types of EB, and the level of expression of these genes within the cutaneous basement membrane zone and in extracutaneous tissues, as well as the types and combinations of the mutations, explain in general terms the phenotypic variability. METHODS: The DebRA Molecular Diagnostics Laboratory, established in 1996 and supported in part by the patient advocacy organisation DebRA of America, has analysed over 1000 families with different forms of EB. RESULTS: In total, 265 cases were submitted with the preliminary diagnosis of junctional or hemidesmosomal forms of EB. We found 393 mutant alleles in seven different genes, with 173 of the mutations being distinct and 71 previously unpublished. DISCUSSION: These findings attest to the clinical and molecular heterogeneity of the junctional and hemidesmosomal subtypes of EB. The results also reveal exceptions to the general rules on genotype-phenotype correlations, unusual phenotypes, and surprising genetics. Collectively, mutation analysis in different forms of EB provides the basis for improved classification with prognostic implications and for prenatal and preimplantation diagnosis in families at risk for recurrence of EB. PMID- 16473854 TI - Poly(ADP-RIBOSE) polymerase-1 (Parp-1) antagonizes topoisomerase I-dependent recombination stimulation by P53. AB - PARP-1 interacts with and poly(ADP-ribosyl)ates p53 and topoisomerase I, which both participate in DNA recombination. Previously, we showed that PARP-1 downregulates homology-directed double-strand break (DSB) repair. We also discovered that, despite the well-established role of p53 as a global suppressor of error-prone recombination, p53 enhances homologous recombination (HR) at the RARalpha breakpoint cluster region (bcr) comprising topoisomerase I recognition sites. Using an SV40-based assay and isogenic cell lines differing in the p53 and PARP-1 status we demonstrate that PARP-1 counteracts HR enhancement by p53, although DNA replication was largely unaffected. When the same DNA element was integrated in an episomal recombination plasmid, both p53 and PARP-1 exerted anti recombinogenic rather than stimulatory activities. Strikingly, with DNA substrates integrated into cellular chromosomes, enhancement of HR by p53 and antagonistic PARP-1 action was seen, very similar to the HR of viral minichromosomes. siRNA-mediated knockdown revealed the essential role of topoisomerase I in this regulatory mechanism. However, after I-SceI-meganuclease mediated cleavage of the chromosomally integrated substrate, no topoisomerase I dependent effects by p53 and PARP-1 were observed. Our data further indicate that PARP-1, probably through topoisomerase I interactions rather than poly(ADP ribosyl)ation, prevents p53 from stimulating spontaneous HR on chromosomes via topoisomerase I activity. PMID- 16473857 TI - Barriers to implementing a policy not to attempt resuscitation in acute medical admissions: prospective, cross sectional study of a successive cohort. AB - OBJECTIVE: To establish whether acutely unwell patients admitted to hospital wish to participate in discussions about resuscitation. DESIGN: Prospective, cross sectional study of a successive cohort of patients. SETTING: Admission through the emergency department. PARTICIPANTS: 374 adult patients. MAIN OUTCOME MEASURE: Whether acutely unwell patients wished to participate in discussions about resuscitation. RESULTS: Of the total sample, 74 patients consented to take part in the study and provide full data. Of the remaining patients, 189 could not be approached for practical reasons and 111 did not wish to participate. Of the 74 patients who read the leaflet, 65 (88%) reported having little or no prior knowledge, 70 (96%) understood it, 56 (77.8%) preferred for resuscitation decisions to be discussed with them, and 55 (77.5%) did not mind discussing resuscitation within 24 hours of admission and overall showed a decline in their anxiety score. CONCLUSION: Many patients admitted through the emergency department for medical reasons cannot participate in their decision not to attempt resuscitation within 24 hours of admission. Patients who were willing to participate rated the information leaflet that was provided positively. PMID- 16473860 TI - Electrical perception of the 'death message' in Chara: characterization of K+ induced depolarization. AB - When the nodal end of an internodal cell of Chara corallina was subjected to a pulse treatment with artificial cell sap, a depolarization lasted in artificial pond water. This depolarization could also be induced by pulse treatment with KCl solution, indicating that K+ in the artificial cell sap is responsible (K+ induced depolarization). The depolarization was prolonged in the presence of 2 mM KCl and the prolonged depolarization was terminated by supplementing with either CaCl2, MgCl2 or NaCl. These results supported the hypothesis that K+ released from the killed cell is responsible for generation of the wound-induced membrane depolarization. PMID- 16473858 TI - Efficacy and safety of exogenous melatonin for secondary sleep disorders and sleep disorders accompanying sleep restriction: meta-analysis. AB - OBJECTIVE: To conduct a systematic review of the efficacy and safety of exogenous melatonin in managing secondary sleep disorders and sleep disorders accompanying sleep restriction, such as jet lag and shiftwork disorder. DATA SOURCES: 13 electronic databases and reference lists of relevant reviews and included studies; Associated Professional Sleep Society abstracts (1999 to 2003). STUDY SELECTION: The efficacy review included randomised controlled trials; the safety review included randomised and non-randomised controlled trials. QUALITY ASSESSMENT: Randomised controlled trials were assessed by using the Jadad Scale and criteria by Schulz et al, and non-randomised controlled trials by the Downs and Black checklist. DATA EXTRACTION AND SYNTHESIS: One reviewer extracted data and another reviewer verified the data extracted. The inverse variance method was used to weight studies and the random effects model was used to analyse data. MAIN RESULTS: Six randomised controlled trials with 97 participants showed no evidence that melatonin had an effect on sleep onset latency in people with secondary sleep disorders (weighted mean difference -13.2 (95% confidence interval -27.3 to 0.9) min). Nine randomised controlled trials with 427 participants showed no evidence that melatonin had an effect on sleep onset latency in people who had sleep disorders accompanying sleep restriction (-1.0 ( 2.3 to 0.3) min). 17 randomised controlled trials with 651 participants showed no evidence of adverse effects of melatonin with short term use (three months or less). CONCLUSIONS: There is no evidence that melatonin is effective in treating secondary sleep disorders or sleep disorders accompanying sleep restriction, such as jet lag and shiftwork disorder. There is evidence that melatonin is safe with short term use. PMID- 16473859 TI - Risk and protective factors for meningococcal disease in adolescents: matched cohort study. AB - OBJECTIVE: To examine biological and social risk factors for meningococcal disease in adolescents. DESIGN: Prospective, population based, matched cohort study with controls matched for age and sex in 1:1 matching. Controls were sought from the general practitioner. SETTING: Six contiguous regions of England, which represent some 65% of the country's population. PARTICIPANTS: 15-19 year olds with meningococcal disease recruited at hospital admission in six regions (representing 65% of the population of England) from January 1999 to June 2000, and their matched controls. METHODS: Blood samples and pernasal and throat swabs were taken from case patients at admission to hospital and from cases and matched controls at interview. Data on potential risk factors were gathered by confidential interview. Data were analysed by using univariate and multivariate conditional logistic regression. RESULTS: 144 case control pairs were recruited (74 male (51%); median age 17.6). 114 cases (79%) were confirmed microbiologically. Significant independent risk factors for meningococcal disease were history of preceding illness (matched odds ratio 2.9, 95% confidence interval 1.4 to 5.9), intimate kissing with multiple partners (3.7, 1.7 to 8.1), being a university student (3.4, 1.2 to 10) and preterm birth (3.7, 1.0 to 13.5). Religious observance (0.09, 0.02 to 0.6) and meningococcal vaccination (0.12, 0.04 to 0.4) were associated with protection. CONCLUSIONS: Activities and events increasing risk for meningococcal disease in adolescence are different from in childhood. Students are at higher risk. Altering personal behaviours could moderate the risk. However, the development of further effective meningococcal vaccines remains a key public health priority. PMID- 16473861 TI - Neutrophil elastase-initiated EGFR/MEK/ERK signaling counteracts stabilizing effect of autocrine TGF-beta on tropoelastin mRNA in lung fibroblasts. AB - Neutrophil elastase (NE) plays an important role in emphysema, a pulmonary disease associated with excessive elastolysis and ineffective repair of interstitial elastin. Besides its direct elastolytic activity, NE releases soluble epidermal growth factor receptor (EGFR) ligands and initiates EGFR/MEK/ERK signaling to downregulate tropoelastin mRNA in neonatal rat lung fibroblasts (DiCamillo SJ, Carreras I, Panchenko MV, Stone PJ, Nugent MA, Foster JA, and Panchenko MP. J Biol Chem 277: 18938-18946, 2002). We now report that NE downregulates tropoelastin mRNA in the rat fetal lung fibroblast line RFL-6. The tropoelastin mRNA downregulation is preceded by release of EGF-like and TGF-alpha like polypeptides and requires EGFR/MEK/ERK signaling, because it is prevented by the EGFR inhibitor AG1478 and the MEK/ERK uncoupler U0126. Tropoelastin expression in RFL-6 fibroblasts is governed by autocrine TGF-beta signaling, because TGF-beta type I receptor kinase inhibitor or TGF-beta neutralizing antibody dramatically decreases tropoelastin mRNA and protein levels. Half-life of tropoelastin mRNA in RFL-6 cells is >24 h, but it is decreased to approximately 8 h by addition of TGF-beta neutralizing antibody, EGF, TGF-alpha, or NE. Tropoelastin mRNA destabilization by NE, EGF, or TGF-alpha is abolished by AG1478 or U0126. EGF-dependent tropoelastin mRNA downregulation is reversed upon ligand withdrawal, whereas chronic EGF treatment leads to persistent downregulation of tropoelastin mRNA and protein levels and decreases insoluble elastin deposition. We conclude that NE-initiated EGFR/MEK/ERK signaling cascade overrides the autocrine TGF-beta signaling on tropoelastin mRNA stability and, therefore, decreases the elastogenic response in RFL-6 fibroblasts. We hypothesize that persistent EGFR/MEK/ERK signaling could impede the TGF-beta induced elastogenesis/elastin repair in the chronically inflamed, elastase/anti elastase imbalanced lung in emphysema. PMID- 16473862 TI - COX-2-derived prostacyclin protects against bleomycin-induced pulmonary fibrosis. AB - Prostacyclin is one of a number of lipid mediators elaborated from the metabolism of arachidonic acid by the cyclooxygenase (COX) enzymes. This prostanoid is a potent inhibitor of platelet aggregation, and its production by endothelial cells and protective role in the vasculature are well established. In contrast, much less is known regarding the function of this prostanoid in other disease processes. We show here that COX-2-dependent production of prostacyclin plays an important role in the development of fibrotic lung disease, limiting both the development of fibrosis and the consequential alterations in lung mechanics. In stark contrast, loss of prostaglandin E(2) synthesis and signaling through the G(s)-coupled EP2 and EP4 receptors had no effect on the development of disease. These findings suggest that prostacyclin analogs will protect against bleomycin induced pulmonary fibrosis in COX-2(-/-) mice. If such protection is observed, investigation of these agents as a novel therapeutic approach to pulmonary fibrosis in humans may be warranted. PMID- 16473863 TI - Improvement of pulmonary hypoplasia associated with congenital diaphragmatic hernia by in utero CFTR gene therapy. AB - Congenital diaphragmatic hernia (CDH) may be an ideal candidate disease for in utero gene therapy as disrupted fetal lung growth plays a significant role in disease outcome. We previously demonstrated that transient in utero overexpression of CFTR during fetal development resulted in lung epithelial proliferation and differentiation. We hypothesized that gene therapy with CFTR would improve the pulmonary hypoplasia associated with congenital diaphragmatic hernia (CDH). CDH was induced by the herbicide 2,4-dichlorophenyl-4-nitrophyl ether (nitrofen) following maternal ingestion at either 10 or 13 days gestation. In utero gene transfer of the CFTR gene was subsequently performed at 16 days gestation. Examination of the fetuses at 22 days gestation revealed little improvement in the CFTR-treated lungs following induction of hernias with nitrofen at 10 days gestation. However, the CFTR gene treatment significantly improved internal surface area, saccular density, overall saccular number, and amount of saccular air space in the lungs that were treated with nitrofen at 13 days gestation. RT-PCR demonstrated that gene transfer occurred following treatment at 13 days gestation but not in the lungs treated with nitrofen at 10 days gestation, despite gene transfer at the same gestational age (16 days) in both groups. As disruption of lung development correlates with the gestational stage at which nitrofen exposure occurs, these results confirmed previous findings that in utero gene transfer efficiency depends on the stage of lung development. Lung development may be significantly delayed in human CDH to allow for successful gene transfer later in gestation, providing a substantial therapeutic window. PMID- 16473864 TI - NKG2D ligands are expressed on stressed human airway epithelial cells. AB - Immune surveillance of the airways is critical to maintain the integrity and health of the lung. We have identified a family of ligands expressed on the surface of stressed airway epithelial cells whose function is to bind the NKG2D activating receptor found on several pulmonary lymphocytes, including natural killer cells, gammadelta(+) T cells, and CD8(+) T cells. We employed real-time PCR and flow cytometry in normal and transformed airway epithelial cell to demonstrate that major histocompatibility complex class I chain-related (MIC) B and the UL-16 binding protein (ULBP) ligands (ULBP1-4) are ubiquitously expressed at the mRNA level in all cell lines. MICA/B surface expression was present on 70% of transformed cell lines but was undetectable on primary cells. We demonstrate that MICA/B and ULBP 1, 2, 3, and 4 expression is rare or absent on the cell surface of unstimulated normal human bronchial epithelial cells although transcripts and intracellular proteins are present. Normal human bronchial epithelial cells exposed to 0.3 mM hydrogen peroxide exhibit an induction of all ligands examined on the cell surface. Surface expression is independent of changes in transcript level or total cellular protein and is mediated by the ERK family of mitogen-activated protein kinases. The induction of NKG2D ligands on stressed airway epithelial cells represents a potentially important mechanism of immune cell activation in regulation of pulmonary health and disease. PMID- 16473865 TI - Cigarette smoke induces proinflammatory cytokine release by activation of NF kappaB and posttranslational modifications of histone deacetylase in macrophages. AB - Cigarette smoke-mediated oxidative stress induces an inflammatory response in the lungs by stimulating the release of proinflammatory cytokines. Chromatin remodeling due to histone acetylation and deacetylation is known to play an important role in transcriptional regulation of proinflammatory genes. The aim of this study was to investigate the molecular mechanism(s) of inflammatory responses caused by cigarette smoke extract (CSE) in the human macrophage-like cell line MonoMac6 and whether the treatment of these cells with the antioxidant glutathione (GSH) monoethyl ester, or modulation of the thioredoxin redox system, can attenuate cigarette smoke-mediated IL-8 release. Exposure of MonoMac6 cells to CSE (1% and 2.5%) increased IL-8 and TNF-alpha production vs. control at 24 h and was associated with significant depletion of GSH levels associated with increased reactive oxygen species release in addition to activation of NF-kappaB. Inhibition of IKK ablated the CSE-mediated IL-8 release, suggesting that this process is dependent on the NF-kappaB pathway. CSE also reduced histone deacetylase (HDAC) activity and HDAC1, HDAC2, and HDAC3 protein levels. This was associated with posttranslational modification of HDAC1, HDAC2, and HDAC3 protein by nitrotyrosine and aldehyde-adduct formation. Pretreatment of cells with GSH monoethyl ester, but not thioredoxin/thioredoxin reductase, reversed cigarette smoke-induced reduction in HDAC levels and significantly inhibited IL-8 release. Thus cigarette smoke-induced release of IL-8 is associated with activation of NF kappaB via IKK and reduction in HDAC levels/activity in macrophages. Moreover, cigarette smoke-mediated proinflammatory events are regulated by the redox status of the cells. PMID- 16473867 TI - CellML2SBML: conversion of CellML into SBML. AB - CellML and SBML are XML-based languages for storage and exchange of molecular biological and physiological reaction models. They use very similar subsets of MathML to specify the mathematical aspects of the models. CellML2SBML is implemented as a suite of XSLT stylesheets that, when applied consecutively, convert models expressed in CellML into SBML without significant loss of information. The converter is based on the most recent stable versions of the languages (CellML version 1.1; SBML Level 2 Version 1), and the XSLT used in the stylesheets adheres to the XSLT version 1.0 specification. Of all 306 models in the CellML repository in April 2005, CellML2SBML converted 91% automatically into SBML. Minor manual changes to the unit definitions in the originals raised the percentage of successful conversions to 96%. AVAILABILITY: http://sbml.org/software/cellml2sbml/. SUPPLEMENTARY INFORMATION: Instructions for use and further documentation available on http://sbml.org/software/cellml2sbml/ PMID- 16473866 TI - Role of cytokinin and auxin in shaping root architecture: regulating vascular differentiation, lateral root initiation, root apical dominance and root gravitropism. AB - BACKGROUND AND AIMS: Development and architecture of plant roots are regulated by phytohormones. Cytokinin (CK), synthesized in the root cap, promotes cytokinesis, vascular cambium sensitivity, vascular differentiation and root apical dominance. Auxin (indole-3-acetic acid, IAA), produced in young shoot organs, promotes root development and induces vascular differentiation. Both IAA and CK regulate root gravitropism. The aims of this study were to analyse the hormonal mechanisms that induce the root's primary vascular system, explain how differentiating-protoxylem vessels promote lateral root initiation, propose the concept of CK-dependent root apical dominance, and visualize the CK and IAA regulation of root gravitropiosm. KEY ISSUES: The hormonal analysis and proposed mechanisms yield new insights and extend previous concepts: how the radial pattern of the root protoxylem vs. protophloem strands is induced by alternating polar streams of high IAA vs. low IAA concentrations, respectively; how differentiating-protoxylem vessel elements stimulate lateral root initiation by auxin-ethylene-auxin signalling; and how root apical dominance is regulated by the root-cap-synthesized CK, which gives priority to the primary root in competition with its own lateral roots. CONCLUSIONS: CK and IAA are key hormones that regulate root development, its vascular differentiation and root gravitropism; these two hormones, together with ethylene, regulate lateral root initiation. PMID- 16473868 TI - Development and validation of a consistency based multiple structure alignment algorithm. AB - SUMMARY: We introduce an algorithm that uses the information gained from simultaneous consideration of an entire group of related proteins to create multiple structure alignments (MSTAs). Consistency-based alignment (CBA) first harnesses the information contained within regions that are consistently aligned among a set of pairwise superpositions in order to realign pairs of proteins through both global and local refinement methods. It then constructs a multiple alignment that is maximally consistent with the improved pairwise alignments. We validate CBA's alignments by assessing their accuracy in regions where at least two of the aligned structures contain the same conserved sequence motif. RESULTS: CBA correctly aligns well over 90% of motif residues in superpositions of proteins belonging to the same family or superfamily, and it outperforms a number of previously reported MSTA algorithms. PMID- 16473869 TI - Comparison of P-RnaPredict and mfold--algorithms for RNA secondary structure prediction. AB - MOTIVATION: Ribonucleic acid is vital in numerous stages of protein synthesis; it also possesses important functional and structural roles within the cell. The function of an RNA molecule within a particular organic system is principally determined by its structure. The current physical methods available for structure determination are time-consuming and expensive. Hence, computational methods for structure prediction are sought after. The energies involved by the formation of secondary structure elements are significantly greater than those of tertiary elements. Therefore, RNA structure prediction focuses on secondary structure. RESULTS: We present P-RnaPredict, a parallel evolutionary algorithm for RNA secondary structure prediction. The speedup provided by parallelization is investigated with five sequences, and a dramatic improvement in speedup is demonstrated, especially with longer sequences. An evaluation of the performance of P-RnaPredict in terms of prediction accuracy is made through comparison with 10 individual known structures from 3 RNA classes (5S rRNA, Group I intron 16S rRNA and 16S rRNA) and the mfold dynamic programming algorithm. P-RnaPredict is able to predict structures with higher true positive base pair counts and lower false positives than mfold on certain sequences. AVAILABILITY: P-RnaPredict is available for non-commercial usage. Interested parties should contact Kay C. Wiese (wiese@cs.sfu.ca). PMID- 16473870 TI - Composite Module Analyst: a fitness-based tool for identification of transcription factor binding site combinations. AB - MOTIVATION: Functionally related genes involved in the same molecular-genetic, biochemical or physiological process are often regulated coordinately. Such regulation is provided by precisely organized binding of a multiplicity of special proteins [transcription factors (TFs)] to their target sites (cis elements) in regulatory regions of genes. Cis-element combinations provide a structural basis for the generation of unique patterns of gene expression. RESULTS: Here we present a new approach for defining promoter models based on the composition of TF binding sites and their pairs. We utilize a multicomponent fitness function for selection of the promoter model that fits best to the observed gene expression profile. We demonstrate examples of successful application of the fitness function with the help of a genetic algorithm for the analysis of functionally related or co-expressed genes as well as testing on simulated and permutated data. AVAILABILITY: The CMA program is freely available for non-commercial users. URL http://www.gene regulation.com/pub/programs.html#CMAnalyst. It is also a part of the commercial system ExPlain (www.biobase.de) designed for causal analysis of gene expression data.. PMID- 16473871 TI - Generation of 3D templates of active sites of proteins with rigid prosthetic groups. AB - MOTIVATION: With the increasing availability of protein structures, the generation of biologically meaningful 3D patterns from the simultaneous alignment of several protein structures is an exciting prospect: active sites could be better understood, protein functions and protein 3D structures could be predicted more accurately. Although patterns can already be generated at the fold and topological levels, no system produces high-resolution 3D patterns including atom and cavity positions. To address this challenge, our research focuses on generating patterns from proteins with rigid prosthetic groups. Since these groups are key elements of protein active sites, the generated 3D patterns are expected to be biologically meaningful. RESULTS: In this paper, we present a new approach which allows the generation of 3D patterns from proteins with rigid prosthetic groups. Using 237 protein chains representing proteins containing porphyrin rings, our method was validated by comparing 3D templates generated from homologues with the 3D structure of the proteins they model. Atom positions were predicted reliably: 93% of them had an accuracy of 1.00 A or less. Moreover, similar results were obtained regarding chemical group and cavity positions. Results also suggested our system could contribute to the validation of 3D protein models. Finally, a 3D template was generated for the active site of human cytochrome P450 CYP17, the 3D structure of which is unknown. Its analysis showed that it is biologically meaningful: our method detected the main patterns of the cytochrome P450 superfamily and the motifs linked to catalytic reactions. The 3D template also suggested the position of a residue, which could be involved in a hydrogen bond with CYP17 substrates and the shape and location of a cavity. Comparisons with independently generated 3D models comforted these hypotheses. AVAILABILITY: Alignment software (Nestor3D) is available at http://www.kingston.ac.uk/~ku33185/Nestor3D.html PMID- 16473872 TI - CFinder: locating cliques and overlapping modules in biological networks. AB - Most cellular tasks are performed not by individual proteins, but by groups of functionally associated proteins, often referred to as modules. In a protein association network modules appear as groups of densely interconnected nodes, also called communities or clusters. These modules often overlap with each other and form a network of their own, in which nodes (links) represent the modules (overlaps). We introduce CFinder, a fast program locating and visualizing overlapping, densely interconnected groups of nodes in undirected graphs, and allowing the user to easily navigate between the original graph and the web of these groups. We show that in gene (protein) association networks CFinder can be used to predict the function(s) of a single protein and to discover novel modules. CFinder is also very efficient for locating the cliques of large sparse graphs. AVAILABILITY: CFinder (for Windows, Linux and Macintosh) and its manual can be downloaded from http://angel.elte.hu/clustering. SUPPLEMENTARY INFORMATION: Supplementary data are available on Bioinformatics online. PMID- 16473873 TI - Non-linear tests for identifying differentially expressed genes or genetic networks. AB - MOTIVATION: One of the recently developed statistics for identifying differentially expressed genetic networks is Hotelling T2 statistic, which is a quadratic form of difference in linear functions of means of gene expressions between two types of tissue samples, and so their power is limited. RESULTS: To improve the power of test statistics, a general statistical framework for construction of non-linear tests is presented, and two specific non-linear test statistics that use non-linear transformations of means are developed. Asymptotical distributions of the non-linear test statistics under the null and alternative hypothesis are derived. It has been proved that under some conditions the power of the non-linear test statistics is higher than that of the T2 statistic. Besides theory, to evaluate in practice the performance of the non linear test statistics, they are applied to two real datasets. The preliminary results demonstrate that the P-values of the non-linear statistics for testing differential expressions of the genetic networks are much smaller than those of the T2 statistic. And furthermore simulations show the Type I errors of the non linear statistics agree with the threshold used and the statistics fit the chi2 distribution. SUPPLEMENTARY INFORMATION: Supplementary data are available on Bioinformatics online. PMID- 16473874 TI - A new summarization method for Affymetrix probe level data. AB - MOTIVATION: We propose a new model-based technique for summarizing high-density oligonucleotide array data at probe level for Affymetrix GeneChips. The new summarization method is based on a factor analysis model for which a Bayesian maximum a posteriori method optimizes the model parameters under the assumption of Gaussian measurement noise. Thereafter, the RNA concentration is estimated from the model. In contrast to previous methods our new method called 'Factor Analysis for Robust Microarray Summarization (FARMS)' supplies both P-values indicating interesting information and signal intensity values. RESULTS: We compare FARMS on Affymetrix's spike-in and Gene Logic's dilution data to established algorithms like Affymetrix Microarray Suite (MAS) 5.0, Model Based Expression Index (MBEI), Robust Multi-array Average (RMA). Further, we compared FARMS with 43 other methods via the 'Affycomp II' competition. The experimental results show that FARMS with default parameters outperforms previous methods if both sensitivity and specificity are simultaneously considered by the area under the receiver operating curve (AUC). We measured two quantities through the AUC: correctly detected expression changes versus wrongly detected (fold change) and correctly detected significantly different expressed genes in two sets of arrays versus wrongly detected (P-value). Furthermore FARMS is computationally less expensive then RMA, MAS and MBEI. AVAILABILITY: The FARMS R package is available from http://www.bioinf.jku.at/software/farms/farms.html. SUPPLEMENTARY INFORMATION: http://www.bioinf.jku.at/publications/papers/farms/supplementary.ps PMID- 16473875 TI - Look-Align: an interactive web-based multiple sequence alignment viewer with polymorphism analysis support. AB - We have developed Look-Align, an interactive web-based viewer to display pre computed multiple sequence alignments. Although initially developed to support the visualization needs of the maize diversity website Panzea (http://www.panzea.org), the viewer is a generic stand-alone tool that can be easily integrated into other websites. AVAILABILITY: Look-Align is written in Perl using open-source components and is available under an open-source license. Live installation and download information can be found at the Panzea website (http://www.panzea.org/software/alignment_viewer.html). CONTACT: ware@cshl.edu SUPPLEMENTARY INFORMATION: The Supplementary information includes sample lists of multiple sequence alignment software and sample screenshots of the viewer. PMID- 16473876 TI - A novel hematopoietic granulin induces proliferation of goldfish (Carassius auratus L.) macrophages. AB - Granulins are a group of highly conserved growth factors that have been described from a variety of organisms spanning the metazoa. In this study, goldfish granulin was one of the most commonly identified transcripts in the differential cross-screening of macrophage cDNA libraries and was preferentially expressed in proliferating macrophages. Unlike mammalian granulins, which possess 7.5 repeats of a characteristic signature of 12 cysteine residues, the goldfish granulin encoded a putative peptide possessing only 1.5 cysteine repeats. Northern blot and real-time PCR analyses indicated that goldfish granulin was expressed only in the hematopoietic tissues of the goldfish, specifically the kidney and spleen, and in activated peripheral blood mononuclear cells. We expressed granulin using a prokaryotic expression system and produced an affinity-purified rabbit anti goldfish granulin IgG. Recombinant goldfish granulin induced a dose-dependent proliferative response of goldfish macrophages that was inversely related to the myeloid differentiation stage of the cells studied. The highest proliferative response was observed in macrophage progenitor cells and monocytes. This proliferative response of macrophages was abrogated by the addition of anti granulin IgG. These results indicate that goldfish granulin is a growth factor that positively modulates cell proliferation at distinct junctures of macrophage differentiation. PMID- 16473877 TI - Ggamma13 interacts with PDZ domain-containing proteins. AB - The G protein gamma13 subunit (Ggamma13) is expressed in taste and retinal and neuronal tissues and plays a key role in taste transduction. We identified PSD95, Veli-2, and other PDZ domain-containing proteins as binding partners for Ggamma13 by yeast two-hybrid and pull-down assays. In two-hybrid assays, Ggamma13 interacted specifically with the third PDZ domain of PSD95, the sole PDZ domain of Veli-2, and the third PDZ domain of SAP97, a PSD95-related protein. Ggamma13 did not interact with the other PDZ domains of PSD95. Coexpression of Ggamma13 with its Gbeta1 partner did not interfere with these two-hybrid interactions. The physical interaction of Ggamma13 with PSD95 in the cellular milieu was confirmed in pull-down assays following heterologous expression in HEK293 cells. The interaction of Ggamma13 with the PDZ domain of PSD95 was via the C-terminal CAAX tail of Ggamma13 (where AA indicates the aliphatic amino acid); alanine substitution of the CTAL sequence at the C terminus of Ggamma13 abolished its interactions with PSD95 in two-hybrid and pull-down assays. Veli-2 and SAP97 were identified in taste tissue and in Ggamma13-expressing taste cells. Coimmunoprecipitation of Ggamma13 and PSD95 from brain and of Ggamma13 and SAP97 from taste tissue indicates that Ggamma13 interacts with these proteins endogenously. This is the first demonstration that PDZ domain proteins interact with heterotrimeric G proteins via the CAAX tail of Ggamma subunits. The interaction of Ggamma13 with PDZ domain-containing proteins may provide a means to target particular Gbetagamma subunits to specific subcellular locations and/or macromolecular complexes involved in signaling pathways. PMID- 16473878 TI - A weak Fe-O bond in the oxygenated complex of the nitric-oxide synthase of Staphylococcus aureus. AB - Little is known about the intermediates formed during catalysis by nitric-oxide synthase (NOS). We report here the characterization by resonance Raman spectroscopy of the oxygenated complex of the NOS from Staphylococcus aureus (saNOS) as well as the kinetics of formation and decay of the complex. An oxygenated complex transiently formed after mixing reduced saNOS with oxygen and decayed to the ferric enzyme with kinetics that were dependent on the substrate L arginine and the cofactor H(4)B. The oxygenated complex displayed a Soret absorption band centered at 430 nm. Resonance Raman spectroscopy revealed that it can be described as a ferric superoxide form (Fe(III)O(2)(-)) with a single nu(O O) mode at 1135 cm(-1). In the presence of L-arginine, an additional nu(O-O) mode at 1123 cm(-1) was observed, indicating an increased pi back-bonding electron donation to the bound oxygen induced by the substrate. With saNOS, this is the first time that the nu(Fe-O) mode of a NOS has been observed. The low frequency of this mode, at 517 cm(-1), points to an oxygenated complex that differs from that of P450(cam). The electronic structure of the oxygenated complex and the effect of L-arginine are discussed in relation to the kinetic properties of saNOS and other NOS. PMID- 16473879 TI - T-bet binding to newly identified target gene promoters is cell type-independent but results in variable context-dependent functional effects. AB - Recently developed target gene identification strategies based upon the chromatin immunoprecipitation assay provide a powerful method to determine the localization of transcription factor binding within mammalian genomes. However, in many cases, it is unclear if the binding capacity of a transcription factor correlates with an obligate role in gene regulation in diverse contexts. It is therefore important to carefully examine the relationship between transcription factor binding and its ability to functionally regulate gene expression. T-bet is a T box transcription factor expressed in several hematopoietic cell types. By utilizing a chromatin immunoprecipitation assay coupled to genomic microarray technology approach, we identified numerous promoters, including CXCR3, IL2Rbeta, and CCL3, that are bound by T-bet in B cells. Most surprisingly, the ability of T bet to associate with the target promoters is not dependent upon the cell type background. Several of the promoters appear to be functionally regulated by T bet. However, we could not detect a functional consequence for T-bet association with many of the identified promoters in overexpression studies or an examination of wild type and T-bet-/- primary B, CD4+, and CD8+ T cells. Thus, there is a high variability in the functional consequences, if any, that result from the association of T-bet with individual target promoters. PMID- 16473880 TI - Molecular properties of Oep21, an ATP-regulated anion-selective solute channel from the outer chloroplast membrane. AB - The flux of phosphorylated carbohydrates, the major export products of chloroplasts, is regulated at the level of the inner and presumably also at the level of the outer membrane. This is achieved through modulation of the outer membrane Oep21 channel currents and tuning of its ion selectivity. Refined analysis of the Oep21 channel properties by biochemical and electrophysiological methods revealed a channel formed by eight beta-strands with a wider pore vestibule of dvest approximately 2.4 nm at the intermembrane site and a narrower filter pore of drestr approximately 1 nm. The Oep21 pore contains two high affinity sites for ATP, one located at a relative transmembrane electrical distance delta = 0.56 and the second close to the vestibule at the intermembrane site. The ATP-dependent current block and reduction in anion selectivity of the Oep21 channel is relieved by the competitive binding of phosphorylated metabolic intermediates like 3-phosphoglycerate and glycerinaldehyde 3-phosphate. Deletion of a C-terminal putative FX4K binding motif in Oep21 decreased the capability of the channel to tune its ion selectivity by about 50%, whereas current block remained unchanged. PMID- 16473881 TI - Crystal structure of the sugar binding domain of the archaeal transcriptional regulator TrmB. AB - TrmB is an alpha-glucoside-sensing transcriptional regulator controlling two operons encoding maltose/trehalose and maltodextrin ABC transporters of Pyrococcus furiosus. The crystal structure of an N-terminal truncated derivative of TrmB (amino acids 2-109 deleted; TrmB(delta2-109)) was solved at 1.5 A resolution. This protein has lost its DNA binding domain but has retained its sugar recognition site. The structure represents a novel sugar-binding fold. TrmB(delta2-109) bound maltose, glucose, sucrose, and maltotriose, exhibiting Kd values of 6.8, 25, 34, and 160 microM, respectively. TrmB(delta2-109) behaved as a monomer in dilute buffer solution in contrast to the full-length protein, which is a dimer. Co-crystallization with bound maltose identified a binding site involving seven amino acid residues: Ser229, Asn305, Gly320, Met321, Val324, Ile325, and Glu326. Six of these residues interact with the nonreducing glucosyl residue of maltose. The nonreducing glucosyl residue is shared by all substrates bound to TrmB, suggesting it as a common recognition motif. PMID- 16473882 TI - Structural definition of the H-2Kd peptide-binding motif. AB - Classic major histocompatibility complex (MHC) proteins associate with antigen- and self-derived peptides in an allele-specific manner. Herein we present the crystal structure of the MHC class I protein H-2K(d) (K(d)) expressed by BALB/c mice in complex with an antigenic peptide derived from influenza A/PR/8/34 nucleoprotein (Flu, residues 147-155, TYQRTRALV). Analysis of our structure in conjunction with the sequences of naturally processed epitopes provides a comprehensive understanding of the dominant K(d) peptide-binding motif. We find that Flu residues Tyr(P2), Thr(P5), and Val(P9) are sequestered into the B, C, and F pockets of the K(d) groove, respectively. The shape and chemistry of the polymorphic B pocket make it an optimal binding site for the side chain of Tyr(P2) as the dominant anchoring residue of nonameric peptides. The non-polar F pocket limits the amino acid repertoire at P9 to hydrophobic residues such as Ile, Leu, or Val, whereas the C pocket restricts the size of the P5-anchoring side chain. We also show that Flu is accommodated in the complex through an unfavorable kink in the otherwise extended peptide backbone due to the presence of a prominent ridge in the K(d) groove. Surprisingly, this backbone conformation is strikingly similar to D(b)-presented peptides despite the fact that these proteins employ distinct motif-anchoring strategies. The results presented in this study provide a solid foundation for the understanding of K(d)-restricted antigen presentation and recognition events. PMID- 16473883 TI - The comparative roles of suppressor of cytokine signaling-1 and -3 in the inhibition and desensitization of cytokine signaling. AB - Negative feedback is a mechanism commonly employed in biological processes as a means of maintaining homeostasis. We have investigated the roles of suppressor of cytokine signaling (SOCS) proteins in regulating the kinetics of negative feedback in response to cytokine signaling. In mouse livers and bone marrow derived macrophages, both interferon-gamma (IFNgamma) and interleukin-6 (IL-6) rapidly induced the tyrosine phosphorylation of signal transducer and activator of transcription-1 (STAT1) and STAT3. STAT3 tyrosine phosphorylation was bi phasic in response to continuous IL-6 signaling. In macrophages lacking Socs3, however, continuous IL-6 signaling induced uniformly high levels of STAT3 tyrosine phosphorylation, and early IL-6-inducible genes were inappropriately expressed at intermediate time points. SOCS3 therefore imposes bi-phasic kinetics upon IL-6 signaling. Compared with Socs3 mRNA, Socs1 mRNA was induced relatively slowly, and SOCS1 simply attenuated the duration of IFNgamma signaling. Surprisingly, heightened Socs1 mRNA expression but minimal STAT1 tyrosine phosphorylation was observed after prolonged stimulation with IFNgamma, indicating that STAT1 may not play a large role in inducing Socs1 mRNA during steady-state IFNgamma signaling. We also demonstrate that both SOCS1 and SOCS3 can desensitize primary bone marrow-derived macrophages to IFNgamma and IL-6 signaling, respectively. Consistent with the kinetics with which Socs1 and Socs3 mRNAs were induced, SOCS3 desensitized cells to IL-6 rapidly, whereas SOCS1 mediated desensitization to IFNgamma occurred at later time points. The kinetics with which SOCS proteins are induced by cytokine may therefore be a parameter that is "hard-wired" into specific cytokine signaling pathways as a means of tailoring the kinetics with which cells become desensitized. PMID- 16473884 TI - Direct stimulation of transcription initiation by BRCA1 requires both its amino and carboxyl termini. AB - Published experiments suggest that BRCA1 interaction with RNAPII and regulation of a number of target genes may be central to its role as a tumor suppressor. Previous in vivo and in vitro work has implicated the carboxyl terminus of BRCA1 in transcriptional stimulation, but the mechanism of action remains unknown, and whether the full-length protein stimulates transcription is controversial. BRCA1 interacts with a number of enhancer-binding transcriptional activators, suggesting that these factors recruit BRCA1 to promoters, where it stimulates RNA synthesis. To investigate whether BRCA1 has intrinsic transcriptional activity, we established a fully purified transcription assay. We demonstrate here that BRCA1 stimulates transcription initiation across a range of promoters. Both the amino and carboxyl termini of BRCA1 are required for this activity, but the BRCA1 binding partner, BARD1, is not. Our data support a model whereby BRCA1 stabilizes productive preinitiation complexes and thus stimulates transcription. PMID- 16473885 TI - Heme oxygenase-1 is essential for and promotes tolerance to transplanted organs. AB - This investigation focused on obtaining a further understanding of the role of heme oxygenase-1 (HO-1) in tolerance induction. Hearts from C57BL/6 (H-2b) mice survived long-term when transplanted into BALB/c (H-2d) recipients treated with the tolerance-inducing regimen of anti-CD40L antibody (MR-1) plus donor-specific transfusion (DST). Grafts did not, however, survive long-term in (HO-1-/-) recipients given the same treatment. Similarly, long-term survival induced by DST was ablated when HO-1 activity was blocked by zinc protoporphyrin IX (ZnPPIX). We further asked whether modulation of HO-1 expression/activity could be used to promote the induction of graft tolerance. DST alone (day 0) failed to promote any prolongation of survival of DBA/2 (H-2d) hearts transplanted into B6AF1 (H 2(b,k/d)) recipients. However, long-term survival and (dominant peripheral) tolerance were readily induced when DST was combined with induction of HO-1 expression by cobalt protoporphyrin IX (CoPPIX). HO-1 induction plus DST led to a significant up-regulation of Foxp3, TGF-beta, IL-10, and CTLA4, which suggests a prominent role for CD4+CD25+ regulatory T cells (Tregs). In fact, the tolerogenic effect of HO-1 plus DST was dependent on CD4+CD25+ Tregs as suggested by adoptively transferring these cells into irradiated recipients under various regimens. Taken together, these findings show that expression of HO-1 in a graft recipient can be essential for long-term graft survival and for induction of tolerance and that modulation of HO-1 expression/activity can be used therapeutically to synergize in the generation of graft tolerance. PMID- 16473886 TI - Vasohibin is up-regulated by VEGF in the retina and suppresses VEGF receptor 2 and retinal neovascularization. AB - Vasohibin is a recently identified protein that is up-regulated in cultured vascular endothelial cells by vascular endothelial growth factor and fibroblast growth factor 2. It inhibits endothelial cell migration, proliferation, and tube formation, and suppresses angiogenesis in chick chorioallantoic membrane, after subcutaneous implantation of matrigel, and in a tumor xenograft model. This has led to the hypothesis that vasohibin functions as a negative feedback inhibitor of angiogenesis. In this study, we tested that hypothesis in a well-characterized model of retinal neovascularization. In ischemic retina, increased expression of VEGF was accompanied by elevation of vasohibin mRNA and blocking of the increase in vegf mRNA with vegf siRNA significantly attenuated the rise in vasohibin mRNA. In transgenic mice in which the rhodopsin promoter drives expression of VEGF in the retina, there was also a significant increase in vasohibin mRNA. In mice with ischemic retinopathy, there was increased expression of vasohibin in vascular endothelial cells, and vasohibin knockdown caused an increase in neovascularization. Conversely, intraocular injection of recombinant vasohibin or an adenoviral vector containing a vasohibin expression cassette strongly suppressed retinal neovascularization in mice with ischemic retinopathy. Knockdown of vasohibin mRNA in ischemic retina had no significant effect on vegf or vegf receptor 1 mRNA levels but caused a significant elevation in the level of vegf receptor 2 mRNA. These data support the hypothesis that vasohibin acts as a negative feedback regulator of neovascularization in the retina and suggest that suppression of VEGF receptor 2 may play some role in mediating its activity. PMID- 16473887 TI - Microglia provide neuroprotection after ischemia. AB - Many neurological insults are accompanied by a marked acute inflammatory reaction, involving the activation of microglia. Using a model of exogenous application of fluorescence-labeled BV2 microglia in pathophysiologically relevant concentrations onto organotypic hippocampal slice cultures, we investigated the specific effects of microglia on neuronal damage after ischemic injury. Neuronal cell death after oxygen-glucose deprivation (OGD) was determined by propidium iodide incorporation and Nissl staining. Migration and interaction with neurons were analyzed by time resolved 3-D two-photon microscopy. We show that microglia protect against OGD-induced neuronal damage and engage in close physical cell-cell contact with neurons in the damaged brain area. Neuroprotection and migration of microglia were not seen with integrin regulator CD11a-deficient microglia or HL-60 granulocytes. The induction of migration and neuron-microglia interaction deep inside the slice was markedly increased under OGD conditions. Lipopolysaccharide-prestimulated microglia failed to provide neuroprotection after OGD. Pharmacological interference with microglia function resulted in a reduced neuroprotection. Microglia proved to be neuroprotective even when applied up to 4 h after OGD, thus defining a "protective time window." In acute injury such as trauma or stroke, appropriately activated microglia may primarily have a neuroprotective role. Anti-inflammatory treatment within the protective time window of microglia would therefore be counterintuitive. PMID- 16473888 TI - Physical and chemical interactions between aphids and plants. AB - Aphids feed from sieve tubes deep inside the host plant. Therefore, aphids must be able to recognize their host plant(s) and to direct their stylets which must be long and thin enough to reach and puncture the sieve tubes at a particular site. Sieve tubes in angiosperms are longitudinal arrays of sieve element/companion cell modules which are highly sensitive to disturbance of any kind. The sieve tubes dispose of elaborate sealing mechanisms such as protein plugging and callose sealing which are triggered by a rise in calcium in the sieve tubes. Aphids seem to have developed a range of physical and chemical measures to limit the amount of calcium influx in response to stylet puncturing. Loss of sieve-element turgor pressure induced by stylet insertion is minimized by the minute stylet volume. Turgor-dependent Ca(2+) influx, possibly mediated by mechano sensitive Ca(2+) channels, must therefore be limited. The components of the sheath and watery saliva play a pivotal role in establishing the physical and chemical constraints on the rise of calcium. Most likely, sheath saliva prevents the influx of calcium from the apoplast by sealing the stylet puncture site while watery saliva may prevent plugging and sealing of sieve plates by potential interaction with SE sap ingredients. PMID- 16473889 TI - Mucilage production during the incompatible interaction between Orobanche crenata and Vicia sativa. AB - Orobanche spp. (broomrapes) are holoparasites lacking in chlorophyll and totally dependent on their host for their supply of nutrients. O. crenata is a severe constraint to legumes cultivation and breeding for resistance remains as one of the best available methods of control. However, little is known about the basis of host resistance to broomrapes. It is a multicomponent event, and resistance based on hampering development and necrosis of broomrape tubercles has been reported. In the present work, the formation of mucilage and occlusion of host xylem vessels associated with the death of O. crenata tubercles were studied histologically. Samples of necrotic O. crenata tubercles established on resistant and susceptible vetch genotypes were collected. The samples were fixed, sectioned and stained using different procedures. The sections were observed at the light microscopy level, either under bright field, epi-fluorescence or confocal laser scanning microscopy. A higher proportion of necrotic tubercles was found on the resistant genotype and this was associated with a higher percentage of occluded vessels. Mucilage is composed mainly by carbohydrates (non-esterified pectins) and the presence of polyphenols was also detected. The mucilage and other substances composed by parasite secretions and host-degraded products was found to block host vessels and obstruct the parasite supply channel, being a quantitative defensive response against O. crenata in vetch, and probably also in other legumes and plants. The presence of foreign substances (i.e. parasite secretions) and host-degraded products (i.e. carbohydrates from cell walls) inside host vessels seems to activate this response and leads to xylem occlusion and further death of established Orobanche tubercles. PMID- 16473890 TI - Immunolocalization of FsPK1 correlates this abscisic acid-induced protein kinase with germination arrest in Fagus sylvatica L. seeds. AB - An enzymatically active recombinant protein kinase, previously isolated and characterized in Fagus sylvatica L. dormant seeds (FsPK1), was used to obtain a specific polyclonal antibody against this protein. Immunoblotting and immunohistochemical analysis of FsPK1 protein in beech seeds showed a strong immunostaining in the nucleus of the cells located in the vascular tissue of the embryonic axis corresponding to the future apical meristem of the root. This protein kinase was found to accumulate in the seeds only when embryo growth was arrested by application of ABA, while the protein amount decreased during stratification, previously proved to alleviate dormancy, and no protein was detected at all when seed germination was induced by addition of GA(3). These results indicate that FsPK1 may be involved in the control of the embryo growth mediated by ABA and GAs during the transition from dormancy to germination in Fagus sylvatica seeds. PMID- 16473891 TI - Contribution of omega-3 fatty acid desaturase and 3-ketoacyl-ACP synthase II (KASII) genes in the modulation of glycerolipid fatty acid composition during cold acclimation in birch leaves. AB - Temperate and boreal tree species respond to low positive temperatures (LT) or a shortening of the photoperiod (SD) by inducing cold acclimation. One of the metabolic consequences of cold acclimation is an increase in fatty acid (FA) desaturation in membrane lipids, which allows functional membrane fluidity to be maintained at LT. The molecular mechanisms of FA desaturation were investigated in leaves of birch seedlings (Betula pendula) during cold acclimation. Four genes involved in FA biosynthesis were isolated: a 3-ketoacyl-ACP synthase II gene (BpKASII) involved in the elongation of palmitoyl-ACP to stearoyl-ACP, and three omega-3 FA desaturase genes (BpFAD3, BpFAD7, and BpFAD8) involved in the desaturation of linoleic acid (18:2) to alpha-linolenic acid (18:3). BpFAD7 was the main omega-3 FAD gene expressed in birch leaves, and it was down-regulated by LT under SD conditions. LT induced the expression of BpFAD3 and BpFAD8 and a synchronous increase in 18:3 occurred in glycerolipids. Changes in the photoperiod did not affect the LT-induced increase in 18:3 in chloroplast lipids (MGDG, DGDG, PG), but it modulated the LT response detected in extra chloroplastic lipids (PC, PE, PI, PS). A decrease in the proportion of the 16 carbon FAs in lipids occurred at LT, possibly in relation to the regulation of BpKASII expression at LT. These results suggest that LT affects the whole FA biosynthesis pathway. They support a co-ordinated action of microsomal (BpFAD3) and chloroplast enzymes (BpFAD7, BpFAD8) in determining the level of 18:3 in extra-chloroplastic membranes, and they highlight the importance of dynamic lipid trafficking. PMID- 16473892 TI - Identification of loci affecting flavour volatile emissions in tomato fruits. AB - Fresh tomato fruit flavour is the sum of the interaction between sugars, acids, and a set of approximately 30 volatile compounds synthesized from a diverse set of precursors, including amino acids, lipids, and carotenoids. Some of these volatiles impart desirable qualities while others are negatively perceived. As a first step to identify the genes responsible for the synthesis of flavour-related chemicals, an attempt was made to identify loci that influence the chemical composition of ripe fruits. A genetically diverse but well-defined Solanum pennellii IL population was used. Because S. pennellii is a small green-fruited species, this population exhibits great biochemical diversity and is a rich source of genes affecting both fruit development and chemical composition. This population was used to identify multiple loci affecting the composition of chemicals related to flavour. Twenty-five loci were identified that are significantly altered in one or more of 23 different volatiles and four were altered in citric acid content. It was further shown that emissions of carotenoid derived volatiles were directly correlated with the fruit carotenoid content. Linked molecular markers should be useful for breeding programmes aimed at improving fruit flavour. In the longer term, the genes responsible for controlling the levels of these chemicals will be important tools for understanding the complex interactions that ultimately integrate to provide the unique flavour of a tomato. PMID- 16473893 TI - The significance of amino acids and amino acid-derived molecules in plant responses and adaptation to heavy metal stress. AB - Plants exposed to heavy metals accumulate an array of metabolites, some to high millimolar concentrations. This review deals with N-containing metabolites frequently preferentially synthesized under heavy metal stress such as Cd, Cu, Ni, and Zn. Special focus is given to proline, but certain other amino acids and oligopeptides, as well as betaine, polyamines, and nicotianamine are also addressed. Particularly for proline a large body of data suggests significant beneficial functions under metal stress. In general, the molecules have three major functions, namely metal binding, antioxidant defence, and signalling. Strong correlative and mechanistic experimental evidence, including work with transgenic plants and algae, has been provided that indicates the involvement of metal-induced proline in metal stress defence. Histidine, other amino acids and particularly phytochelatins and glutathione play a role in metal binding, while polyamines function as signalling molecules and antioxidants. Their accumulation needs to be considered as active response and not as consequence of metabolic dys regulation. PMID- 16473894 TI - SECRET AGENT and SPINDLY have overlapping roles in the development of Arabidopsis thaliana L. Heyn. AB - O-GlcNAc transferase (OGT) catalyses transfer of GlcNAc (N-acetylglucosamine) to serine or threonine of proteins. The Arabidopsis OGTs, SECRET AGENT (SEC) and SPINDLY (SPY) have overlapping functions during gametogenesis and embryogenesis. SPY functions in a number of processes including circadian, light, and gibberellin (GA) responses. The role of SEC in plant development and GA signalling was investigated by determining the phenotypes of sec-1 and sec-2 plants and the expression pattern of SEC. Similar to SPY, SEC transcripts were ubiquitous. Although there is no evidence of transcript-level regulation by other factors, SEC mRNA levels are elevated in spy plants and SPY mRNA levels are elevated in sec plants. sec-1 and sec-2 plants exhibited few of the defects observed in spy plants and had wild-type GA responses. Compared with wild type, sec plants produced leaves at a reduced rate. Haplo-insufficiency at SEC in a spy ga1 double mutant background suppressed spy during germination and enhanced the production of ovaries with four carpels by spy. By contrast, SPY haplo insufficiency in a sec ga1 double mutant background caused a novel phenotype, production of a proliferation of pin-like structures instead of a floral shoot. These results are consistent with SEC function overlapping with SPY for leaf production and reproductive development. PMID- 16473895 TI - The plant energy-dissipating mitochondrial systems: depicting the genomic structure and the expression profiles of the gene families of uncoupling protein and alternative oxidase in monocots and dicots. AB - The simultaneous existence of alternative oxidases and uncoupling proteins in plants has raised the question as to why plants need two energy-dissipating systems with apparently similar physiological functions. A probably complete plant uncoupling protein gene family is described and the expression profiles of this family compared with the multigene family of alternative oxidases in Arabidopsis thaliana and sugarcane (Saccharum sp.) employed as dicot and monocot models, respectively. In total, six uncoupling protein genes, AtPUMP1-6, were recognized within the Arabidopsis genome and five (SsPUMP1-5) in a sugarcane EST database. The recombinant AtPUMP5 protein displayed similar biochemical properties as AtPUMP1. Sugarcane possessed four Arabidopsis AOx1-type orthologues (SsAOx1a-1d); no sugarcane orthologue corresponding to Arabidopsis AOx2-type genes was identified. Phylogenetic and expression analyses suggested that AtAOx1d does not belong to the AOx1-type family but forms a new (AOx3-type) family. Tissue-enriched expression profiling revealed that uncoupling protein genes were expressed more ubiquitously than the alternative oxidase genes. Distinct expression patterns among gene family members were observed between monocots and dicots and during chilling stress. These findings suggest that the members of each energy-dissipating system are subject to different cell or tissue/organ transcriptional regulation. As a result, plants may respond more flexibly to adverse biotic and abiotic conditions, in which oxidative stress is involved. PMID- 16473897 TI - Conformational transitions in protein-protein association: binding of fasciculin 2 to acetylcholinesterase. AB - The neurotoxin fasciculin-2 (FAS2) is a picomolar inhibitor of synaptic acetylcholinesterase (AChE). The dynamics of binding between FAS2 and AChE is influenced by conformational fluctuations both before and after protein encounter. Submicrosecond molecular dynamics trajectories of apo forms of fasciculin, corresponding to different conformational substates, are reported here with reference to the conformational changes of loop I of this three fingered toxin. This highly flexible loop exhibits an ensemble of conformations within each substate corresponding to its functions. The high energy barrier found between the two major substates leads to transitions that are slow on the timescale of the diffusional encounter of noninteracting FAS2 and AChE. The more stable of the two apo substates may not be the one observed in the complex with AChE. It seems likely that the more stable apo form binds rapidly to AChE and conformational readjustments then occur in the resulting encounter complex. PMID- 16473896 TI - pH dependence of light-driven proton pumping by an archaerhodopsin from Tibet: comparison with bacteriorhodopsin. AB - The pH-dependence of photocycle of archaerhodopsin 4 (AR4) was examined, and the underlying proton pumping mechanism investigated. AR4 is a retinal-containing membrane protein isolated from a strain of halobacteria from a Tibetan salt lake. It acts as a light-driven proton pump like bacteriorhodopsin (BR). However, AR4 exhibits an "abnormal" feature--the time sequence of proton release and uptake is reversed at neutral pH. We show here that the temporal sequence of AR4 reversed to "normal"--proton release preceding proton uptake--when the pH is increased above 8.6. We estimated the pK(a) of the proton release complex (PRC) in the M intermediate to be approximately 8.4, much higher than 5.7 of wide-type BR. The pH-dependence of the rate constant of M-formation shows that the pK(a) of PRC in the initial state of AR4 is approximately 10.4, whereas it is 9.7 in BR. Thus in AR4, the chromophore photoisomerization and subsequent proton transport from the Schiff base to Asp-85 is coupled to a decrease in the pK(a) of PRC from 10.4 to 8.4, which is 2 pK units less than in BR (4 units). This weakened coupling accounts for the lack of early proton release at neutral pH and the reversed time sequence of proton release and uptake in AR4. Nevertheless the PRC in AR4 effectively facilitates deprotonation of primary proton acceptor and recovery of initial state at neutral pH. We found also that all pK(a)s of the key amino acid residues in AR4 were elevated compared to those of BR. PMID- 16473898 TI - Translocation boost protein-folding efficiency of double-barreled chaperonins. AB - Incorrect folding of proteins in living cells may lead to malfunctioning of the cell machinery. To prevent such cellular disasters from happening, all cells contain molecular chaperones that assist nonnative proteins in folding into the correct native structure. One of the most studied chaperone complexes is the GroEL-GroES complex. The GroEL part has a "double-barrel" structure, which consists of two cylindrical chambers joined at the bottom in a symmetrical fashion. The hydrophobic rim of one of the GroEL chambers captures nonnative proteins. The GroES part acts as a lid that temporarily closes the filled chamber during the folding process. Several capture-folding-release cycles are required before the nonnative protein reaches its native state. Here we report molecular simulations that suggest that translocation of the nonnative protein through the equatorial plane of the complex boosts the efficiency of the chaperonin action. If the target protein is correctly folded after translocation, it is released. However, if it is still nonnative, it is likely to remain trapped in the second chamber, which then closes to start a reverse translocation process. This shuttling back and forth continues until the protein is correctly folded. Our model provides a natural explanation for the prevalence of double-barreled chaperonins. Moreover, we argue that internal folding is both more efficient and safer than a scenario where partially refolded proteins escape from the complex before being recaptured. PMID- 16473899 TI - Determination of the two-dimensional interaction rate constants of a cytokine receptor complex. AB - Ligand-receptor interactions within the plane of the plasma membrane play a pivotal role for transmembrane signaling. The biophysical principles of protein protein interactions on lipid bilayers, though, have hardly been experimentally addressed. We have dissected the interactions involved in ternary complex formation by ligand-induced cross-linking of the subunits of the type I interferon (IFN) receptors ifnar1 and ifnar2 in vitro. The extracellular domains ifnar1-ectodomain (EC) and ifnar2-EC were tethered in an oriented manner on solid supported lipid bilayers. The interactions of IFNalpha2 and several mutants, which exhibit different association and dissociation rate constants toward ifnar1 EC and ifnar2-EC, were monitored by simultaneous label-free detection and surface sensitive fluorescence spectroscopy. Surface dissociation rate constants were determined by measuring ligand exchange kinetics, and by measuring receptor exchange on the surface by fluorescence resonance energy transfer. Strikingly, approximately three-times lower dissociation rate constants were observed for both receptor subunits compared to the dissociation in solution. Based on these directly determined surface-dissociation rate constants, the surface-association rate constants were assessed by probing ligand dissociation at different relative surface concentrations of the receptor subunits. In contrast to the interaction in solution, the association rate constants depended on the orientation of the receptor components. Furthermore, the large differences in association kinetics observed in solution were not detectable on the surface. Based on these results, the key roles of orientation and lateral diffusion on the kinetics of protein interactions in plane of the membrane are discussed. PMID- 16473900 TI - Monolayers of a model anesthetic-binding membrane protein: formation, characterization, and halothane-binding affinity. AB - hbAP0 is a model membrane protein designed to possess an anesthetic-binding cavity in its hydrophilic domain and a cation channel in its hydrophobic domain. Grazing incidence x-ray diffraction shows that hbAP0 forms four-helix bundles that are vectorially oriented within Langmuir monolayers at the air-water interface. Single monolayers of hbAP0 on alkylated solid substrates would provide an optimal system for detailed structural and dynamical studies of anesthetic peptide interaction via x-ray and neutron scattering and polarized spectroscopic techniques. Langmuir-Blodgett and Langmuir-Schaeffer deposition and self-assembly techniques were used to form single monolayer films of the vectorially oriented peptide hbAP0 via both chemisorption and physisorption onto suitably alkylated solid substrates. The films were characterized by ultraviolet absorption, ellipsometry, circular dichroism, and polarized Fourier transform infrared spectroscopy. The alpha-helical secondary structure of the peptide was retained in the films. Under certain conditions, the average orientation of the helical axis was inclined relative to the plane of the substrate, approaching perpendicular in some cases. The halothane-binding affinity of the vectorially oriented hbAP0 peptide in the single monolayers, with the volatile anesthetic introduced into the moist vapor environment of the monolayer, was found to be similar to that for the detergent-solubilized peptide. PMID- 16473901 TI - Reversible B/Z-DNA transition under the low salt condition and non-B-form polydApolydT selectivity by a cubane-like europium-L-aspartic acid complex. AB - We report here that a cubane-like europium-L-aspartic acid complex at physiological pH can discriminate between DNA structures as judged by the comparison of thermal denaturation, binding stoichiometry, temperature-dependent fluorescence enhancement, and circular dichroism and gel electrophoresis studies. This complex can selectively stabilize non-B-form DNA polydApolydT but destabilize polydGdCpolydGdC and polydAdTpolydAdT. Further studies show that this complex can convert B-form polydGdCpolydGdC to Z-form under the low salt condition at physiological temperature 37 degrees C, and the transition is reversible, similar to RNA polymerase, which turns unwound DNA into Z-DNA and converts it back to B-DNA after transcription. The potential uses of a left handed helix-selective probe in biology are obvious. Z-DNA is a transient structure and does not exist as a stable feature of the double helix. Therefore, probing this transient structure with a metal-amino acid complex under the low salt condition at physiological temperature would provide insights into their transitions in vivo and are of great interest. PMID- 16473902 TI - Protein grabs a ligand by extending anchor residues: molecular simulation for Ca2+ binding to calmodulin loop. AB - The structural difference in proteins between unbound and bound forms directly suggests the importance of the conformational plasticity of proteins. However, pathways that connect two-end structures and how they are coupled to the binding reaction are not well understood at atomic resolution. Here, we analyzed the free energy landscape, explicitly taking into account coupling between binding and conformational change by performing atomistic molecular dynamics simulations for Ca2+ binding to a calmodulin loop. Using the AMBER force field with explicit water solvent, we conducted umbrella sampling for the free-energy surface and steered molecular dynamics for the pathway search. We found that, at an early stage of binding, some key residue side chains extend their "arms" to catch Ca2+ and, after catching, they carry the Ca2+ to the center of the binding pocket. This grabbing motion resulted in smooth and stepwise exchange in coordination partners of Ca2+ from water oxygen to atoms in the calmodulin loop. The key residue that first caught the ion was one of the two acidic residues, which are highly conserved. In the pathway simulations, different pathways were observed between binding and dissociation reactions: The former was more diverse than the latter. PMID- 16473903 TI - Forced-unfolding and force-quench refolding of RNA hairpins. AB - Nanomanipulation of individual RNA molecules, using laser optical tweezers, has made it possible to infer the major features of their energy landscape. Time dependent mechanical unfolding trajectories, measured at a constant stretching force (f(S)) of simple RNA structures (hairpins and three-helix junctions) sandwiched between RNA/DNA hybrid handles show that they unfold in a reversible all-or-none manner. To provide a molecular interpretation of the experiments we use a general coarse-grained off-lattice Go-like model, in which each nucleotide is represented using three interaction sites. Using the coarse-grained model we have explored forced-unfolding of RNA hairpin as a function of f(S) and the loading rate (r(f)). The simulations and theoretical analysis have been done both with and without the handles that are explicitly modeled by semiflexible polymer chains. The mechanisms and timescales for denaturation by temperature jump and mechanical unfolding are vastly different. The directed perturbation of the native state by f(S) results in a sequential unfolding of the hairpin starting from their ends, whereas thermal denaturation occurs stochastically. From the dependence of the unfolding rates on r(f) and f(S) we show that the position of the unfolding transition state is not a constant but moves dramatically as either r(f) or f(S) is changed. The transition-state movements are interpreted by adopting the Hammond postulate for forced-unfolding. Forced-unfolding simulations of RNA, with handles attached to the two ends, show that the value of the unfolding force increases (especially at high pulling speeds) as the length of the handles increases. The pathways for refolding of RNA from stretched initial conformation, upon quenching f(S) to the quench force f(Q), are highly heterogeneous. The refolding times, upon force-quench, are at least an order-of magnitude greater than those obtained by temperature-quench. The long f(Q) dependent refolding times starting from fully stretched states are analyzed using a model that accounts for the microscopic steps in the rate-limiting step, which involves the trans to gauche transitions of the dihedral angles in the GAAA tetraloop. The simulations with explicit molecular model for the handles show that the dynamics of force-quench refolding is strongly dependent on the interplay of their contour length and persistence length and the RNA persistence length. Using the generality of our results, we also make a number of precise experimentally testable predictions. PMID- 16473904 TI - Protein-cofactor interactions in bacterial reaction centers from Rhodobacter sphaeroides R-26: I. Identification of the ENDOR lines associated with the hydrogen bonds to the primary quinone QA*-. AB - Hydrogen bonds are important in determining the structure and function of biomolecules. Of particular interest are hydrogen bonds to quinones, which play an important role in the bioenergetics of respiration and photosynthesis. In this work we investigated the hydrogen bonds to the two carbonyl oxygens of the semiquinone QA*- in the well-characterized reaction center from the photosynthetic bacterium Rhodobacter sphaeroides R-26. We used electron paramagnetic resonance and electron nuclear double resonance techniques at 35 GHz at a temperature of 80 K. The goal of this study was to identify and assign sets of 1H-ENDOR lines to protons hydrogen bonded to each of the two oxygens. This was accomplished by preferentially exchanging the hydrogen bond on one of the oxygens with deuterium while concomitantly monitoring the changes in the amplitudes of the 1H-ENDOR lines. The preferential deuteration of one of the oxygens was made possible by the different 1H --> 2H exchange times of the protons bonded to the two oxygens. The assignment of the 1H-ENDOR lines sets the stage for the determination of the geometries of the H-bonds by a detailed field selection ENDOR study to be presented in a future article. PMID- 16473906 TI - Oxygen profiles in membranes. AB - Transmembrane profiles of molecular oxygen in lipid bilayers are not only significant for membrane physiology and pathology, but also are essential to the determination of membrane protein structure by site-directed spin labeling. Oxygen profiles obtained with spin-labeled lipid chains have a Boltzmann sigmoidal dependence on the depth into each lipid leaflet, which represents a two compartment distribution between outer and inner regions of the membrane, with a transfer free energy that depends linearly on distance from the dividing planes. Transmembrane profiles for intramembrane polarity, and for water penetration into the membrane, have an identical form, but are of the reverse sign. Comparison with recently published oxygen profiles from a site-specifically spin-labeled alpha-helical transmembrane peptide validates the use of spin-labeled lipids for all these profiles and provides the necessary bridge to generate the full bilayer from a single lipid leaflet. PMID- 16473907 TI - Force-controlled equilibria of specific vesicle-substrate adhesion. AB - We have developed "vertical" magnetic tweezers capable of exerting controlled pico and subpico Newton forces. Using this apparatus, we apply a point-like force to a vesicle that is adhered by means of specific bonds between the vesicle carrying oligosaccharide sialyl LewisX and the surface-grafted E-selectin. An exponential decrease of the bound vesicle area with the decay rate that is insensitive to the force and the composition of the system is observed. We measure an equilibrium under force that is associated with an increased binding in the center of the contact zone. We also show that the determination of the shape is potentially sufficient to determine the number of formed specific bonds. PMID- 16473905 TI - Catch force links and the low to high force transition of myosin. AB - Catch is characterized by maintenance of force with very low energy utilization in some invertebrate muscles. Catch is regulated by phosphorylation of the mini titin, twitchin, and a catch component of force exists at all [Ca2+] except those resulting in maximum force. The mechanism responsible for catch force was characterized by determining how the effects of agents that inhibit the low to high force transition of the myosin cross-bridge (inorganic phosphate, butanedione monoxime, trifluoperazine, and blebbistatin) are modified by twitchin phosphorylation and [Ca2+]. In permeabilized anterior byssus retractor muscles from Mytilus edulis, catch force was identified as being sensitive to twitchin phosphorylation, whereas noncatch force was insensitive. In all cases, inhibition of the low to high force transition caused an increase in catch force. The same relationship exists between catch force and noncatch force whether force is varied by changes in [Ca2+] and/or agents that inhibit cross-bridge force production. This suggests that myosin in the high force state detaches catch force maintaining structures, whereas myosin in the low force state promotes their formation. It is unlikely that the catch structure is the myosin cross bridge; rather, it appears that myosin interacts with the structure, most likely twitchin, and regulates its attachment and detachment. PMID- 16473908 TI - Onset of anthrax toxin pore formation. AB - Protective antigen (PA) is the anthrax toxin protein recognized by capillary morphogenesis gene 2 (CMG2), a transmembrane cellular receptor. Upon activation, seven ligand-receptor units self-assemble into a heptameric ring-like complex that becomes endocytozed by the host cell. A critical step in the subsequent intoxication process is the formation and insertion of a pore into the endosome membrane by PA. The pore conversion requires a change in binding between PA and its receptor in the acidified endosome environment. Molecular dynamics simulations totaling approximately 136 ns on systems of over 92,000 atoms were performed. The simulations revealed how the PA-CMG2 complex, stable at neutral conditions, becomes transformed at low pH upon protonation of His-121 and Glu 122, two conserved amino acids of the receptor. The protonation disrupts a salt bridge important for the binding stability and leads to the detachment of PA domain II, which weakens the stability of the PA-CMG2 complex significantly, and subsequently releases a PA segment needed for pore formation. The simulations also explain the great strength of the PA-CMG2 complex achieves through extraordinary coordination of a divalent cation. PMID- 16473909 TI - Dissimilar kinetic behavior of electrically manipulated single- and double stranded DNA tethered to a gold surface. AB - We report on the electrical manipulation of single- and double-stranded oligodeoxynucleotides that are end tethered to gold surfaces in electrolyte solution. The response to alternating repulsive and attractive electric surface fields is studied by time-resolved fluorescence measurements, revealing markedly distinct dynamics for the flexible single-stranded and stiff double-stranded DNA, respectively. Hydrodynamic simulations rationalize this finding and disclose two different kinetic mechanisms: stiff polymers undergo rotation around the anchoring pivot point; flexible polymers, on the other hand, are pulled onto the attracting surface segment by segment. PMID- 16473910 TI - Gating of cyclic nucleotide-gated (CNGA1) channels by cGMP jumps and depolarizing voltage steps. AB - We expressed rod-type homotetrameric cyclic nucleotide-gated (CNGA1) channels in Xenopus oocytes and studied activation by photolysis-induced jumps of the 3',5' cyclic guanosine monophosphate (cGMP) concentration and by voltage steps. cGMP jumps to increasing concentrations up to the EC50 value of 46.5 microM decelerate the activation gating, indicative that even at concentrations of cGMP << EC50 binding is not rate limiting. Above the EC50 value, activation by cGMP jumps is again accelerated to the higher concentrations. At the same cGMP concentration, the speed of the activation gating by depolarizing voltage steps is roughly similar to that by cGMP jumps. Permeating ions passing the pore more slowly (Rb+ > K+ > Na+) slow down the activation time course. At the single-channel level, cGMP jumps to high concentrations cause openings directly to the main open level without passing sublevels. From these results it is concluded that at both low and high cGMP the gating of homotetrameric CNGA1 channels is not rate-limited by the cGMP binding but by conformational changes of the channel which are voltage dependent and include movements in the pore region. PMID- 16473912 TI - Numerical simulation of gel electrophoresis of DNA knots in weak and strong electric fields. AB - Gel electrophoresis allows one to separate knotted DNA (nicked circular) of equal length according to the knot type. At low electric fields, complex knots, being more compact, drift faster than simpler knots. Recent experiments have shown that the drift velocity dependence on the knot type is inverted when changing from low to high electric fields. We present a computer simulation on a lattice of a closed, knotted, charged DNA chain drifting in an external electric field in a topologically restricted medium. Using a Monte Carlo algorithm, the dependence of the electrophoretic migration of the DNA molecules on the knot type and on the electric field intensity is investigated. The results are in qualitative and quantitative agreement with electrophoretic experiments done under conditions of low and high electric fields. PMID- 16473911 TI - A dynamic model of excitation-contraction coupling during acidosis in cardiac ventricular myocytes. AB - Acidosis in cardiac myocytes is a major factor in the reduced inotropy that occurs in the ischemic heart. During acidosis, diastolic calcium concentration and the amplitude of the calcium transient increase, while the strength of contraction decreases. This has been attributed to the inhibition by protons of calcium uptake and release by the sarcoplasmic reticulum, to a rise of intracellular sodium caused by activation of sodium-hydrogen exchange, decreased calcium binding affinity to Troponin-C, and direct effects on the contractile machinery. The relative contributions and concerted action of these effects are, however, difficult to establish experimentally. We have developed a mathematical model to examine altered calcium-handling mechanisms during acidosis. Each of the alterations was incorporated into a dynamical model of pH regulation and excitation-contraction coupling to predict the time courses of key ionic species during acidosis, in particular intracellular pH, sodium and the calcium transient, and contraction. This modeling study suggests that the most significant effects are elevated sodium, inhibition of sodium-calcium exchange, and the direct interaction of protons with the contractile machinery; and shows how the experimental data on these contributions can be reconciled to understand the overall effects of acidosis in the beating heart. PMID- 16473913 TI - Measuring the elasticity of clathrin-coated vesicles via atomic force microscopy. AB - Using a new scheme based on atomic force microscopy (AFM), we investigate mechanical properties of clathrin-coated vesicles (CCVs). CCVs are multicomponent protein and lipid complexes of approximately 100 nm diameter that are implicated in many essential cell-trafficking processes. Our AFM imaging resolves clathrin lattice polygons and provides height deformation in quantitative response to AFM substrate compression force. We model CCVs as multilayered elastic spherical shells and, from AFM measurements, estimate their bending rigidity to be 285 +/- 30 k(B)T, i.e., approximately 20 times that of either the outer clathrin cage or inner vesicle membrane. Further analysis reveals a flexible coupling between the clathrin coat and the membrane, a structural property whose modulation may affect vesicle biogenesis and cellular function. PMID- 16473915 TI - Structural analysis of an Echinococcus granulosus actin-fragmenting protein by small-angle x-ray scattering studies and molecular modeling. AB - The Echinococcus granulosus actin filament-fragmenting protein (EgAFFP) is a three domain member of the gelsolin family of proteins, which is antigenic to human hosts. These proteins, formed by three or six conserved domains, are involved in the dynamic rearrangements of the cytoskeleton, being responsible for severing and capping actin filaments and promoting nucleation of actin monomers. Various structures of six domain gelsolin-related proteins have been investigated, but little information on the structure of three domain members is available. In this work, the solution structure of the three domain EgAFFP has been investigated through small-angle x-ray scattering (SAXS) studies. EgAFFP exhibits an elongated molecular shape. The radius of gyration and the maximum dimension obtained by SAXS were, respectively, 2.52 +/- 0.01 nm and 8.00 +/- 1.00 nm, both in the absence and presence of Ca2+. Two different molecular homology models were built for EgAFFP, but only one was validated through SAXS studies. The predicted structure for EgAFFP consists of three repeats of a central beta sheet sandwiched between one short and one long alpha-helix. Possible implications of the structure of EgAFFP upon actin binding are discussed. PMID- 16473914 TI - Immunophysical properties and prediction of activities for vaccinia virus complement control protein and smallpox inhibitor of complement enzymes using molecular dynamics and electrostatics. AB - We present immunophysical modeling for VCP, SPICE, and three mutants using MD simulations and Poisson-Boltzmann-type electrostatic calculations. VCP and SPICE are homologous viral proteins that control the complement system by imitating, structurally and functionally, natural regulators of complement activation. VCP and SPICE consist of four CCP modules connected with short flexible loops. MD simulations demonstrate that the rather complex modules of VCP/SPICE and their mutants exhibit a high degree of intermodular spatial mobility, which is affected by surface mutations. Electrostatic calculations using snapshots from the MD trajectories demonstrate variable spatial distribution of the electrostatic potentials, which suggests dynamic binding properties. We use covariance analysis to identify correlated modular oscillations. We also use electrostatic similarity indices to cluster proteins with common electrostatic properties. Our results are compared with experimental data to form correlations between the overall positive electrostatic potential of VCP/SPICE with binding and activity. We show how these correlations can be used to predict binding and activity properties. This work is expected to be useful for understanding the function of native CCP-containing regulators of complement activation and receptors and for the design of antiviral therapeutics and complement inhibitors. PMID- 16473916 TI - Stochastic kinetics of viral capsid assembly based on detailed protein structures. AB - We present a generic computational framework for the simulation of viral capsid assembly which is quantitative and specific. Starting from PDB files containing atomic coordinates, the algorithm builds a coarse-grained description of protein oligomers based on graph rigidity. These reduced protein descriptions are used in an extended Gillespie algorithm to investigate the stochastic kinetics of the assembly process. The association rates are obtained from a diffusive Smoluchowski equation for rapid coagulation, modified to account for water shielding and protein structure. The dissociation rates are derived by interpreting the splitting of oligomers as a process of graph partitioning akin to the escape from a multidimensional well. This modular framework is quantitative yet computationally tractable, with a small number of physically motivated parameters. The methodology is illustrated using two different viruses which are shown to follow quantitatively different assembly pathways. We also show how in this model the quasi-stationary kinetics of assembly can be described as a Markovian cascading process, in which only a few intermediates and a small proportion of pathways are present. The observed pathways and intermediates can be related a posteriori to structural and energetic properties of the capsid oligomers. PMID- 16473918 TI - Gene expression in human hepatocytes in suspension after isolation is similar to the liver of origin, is not affected by hepatocyte cold storage and cryopreservation, but is strongly changed after hepatocyte plating. AB - Isolated primary human hepatocytes are a well accepted system for evaluating pharmacological and toxicological effects in humans. However, questions remain regarding how culturing affects the liver-specific functions of the hepatocytes. In addition, cryopreservation could also potentially affect the differentiation state of the hepatocytes. The first aim of the present study was to compare gene expression in freshly isolated primary hepatocytes to that of the liver of origin and to evaluate the expression changes occurring after cryopreservation/thawing, both when maintained in suspension and after plating. The second aim of the present study was to evaluate gene expression in hepatocytes after cold storage of suspensions up to 24 h compared with freshly isolated hepatocytes in suspension. Our results show that the gene expression in freshly isolated human hepatocytes in suspension after isolation is similar to that of the liver of origin. Furthermore, gene expression in primary human hepatocytes in suspension is not affected by hepatocyte cold storage and cryopreservation. However, the gene expression is profoundly affected in monolayer cultures after plating. Specifically, gene expression changes were observed in cultured relative to suspensions of human hepatocytes that are involved in cellular processes such as phase I/II metabolism, basolateral and canalicular transport systems, fatty acid and lipid metabolism, apoptosis, and proteasomal protein recycling. An oxidative stress response may be partially involved in these changes in gene expression. Taken together, these results may aid in the interpretation of data collected from human hepatocyte experiments and suggest additional utility for cold storage and cryopreservation of hepatocytes. PMID- 16473917 TI - The canine CYP1A2 deficiency polymorphism dramatically affects the pharmacokinetics of 4-cyclohexyl-1-ethyl-7-methylpyrido[2,3-D]-pyrimidine-2-(1H) one (YM-64227), a phosphodiesterase type 4 inhibitor. AB - In a previous study, it was shown that the novel canine single nucleotide polymorphism (SNP) CYP1A2 1117C>T yields an inactive enzyme. In this study, the effect that this SNP has on the pharmacokinetics of 4-cyclohexyl-1-ethyl-7 methylpyrido[2,3-d]pyrimidine-2-(1H)-one (YM-64227) was investigated. Plasma concentrations of the unchanged drug and five of its metabolites (MM-1 to MM-5) were determined after either intravenous or oral administration of YM-64227 to genotyped dogs (C/C, C/T, and T/T groups). Liver microsomes were prepared from these dogs to determine the in vitro metabolic clearance of YM-64227. After a single oral administration, the maximum plasma concentration and absolute bioavailability of YM-64227 in the T/T group were 17.1 times and 27.2 times higher than those in the C/C group, respectively, whereas the pharmacokinetics of YM-64227 after intravenous administration were not affected by genotype. The metabolic profiles in the T/T group were quite distinct from the others; i.e., the main metabolite was MM-2 in the C/C group, whereas MM-1 and MM-5 were the main metabolites in the T/T group. The formation clearances of MM-2 and MM-3 in the microsomes derived from T/T type dogs were significantly lower, whereas those of MM-1, MM-4, and MM-5 were not affected. A statistically significant correlation was observed between the in vivo and in vitro metabolic intrinsic clearances (r = 0.82, p < 0.001). The canine CYP1A2 1117C>T SNP proved to be responsible for a substantial portion of the interindividual variability in the pharmacokinetics of YM-64227. PMID- 16473919 TI - Inhibition of periodontopathogen-derived proteolytic enzymes by a high-molecular weight fraction isolated from cranberry. AB - BACKGROUND: Porphyromonas gingivalis, Tannerella forsythia and Treponema denticola are three major aetiological agents of chronic periodontitis. The strong proteolytic activities of these bacteria are critical to their survival since their energy source is obtained from peptides and amino acids derived from proteins. In addition, proteases are important factors contributing to periodontal tissue destruction by a variety of mechanisms, including direct tissue degradation and modulation of host inflammatory responses. OBJECTIVES: The aim of this study was to investigate the effect of non-dialysable material (NDM) prepared from cranberry juice concentrate on the proteolytic activities of P. gingivalis, T. forsythia and T. denticola. METHODS: The effect of NDM on gingipain and dipeptidyl peptidase IV (DPP IV) activities of P. gingivalis, trypsin-like activity of T. forsythia and chymotrypsin-like activity of T. denticola was evaluated using synthetic chromogenic peptides. In addition, the capacity of P. gingivalis to degrade fluorescein-labelled type I collagen and fluorescein-labelled transferrin in the presence of NDM was evaluated by fluorometry. RESULTS: NDM dose-dependently inhibited the proteinases of P. gingivalis, T. forsythia and T. denticola as well as type I collagen and transferrin degradation by P. gingivalis. CONCLUSIONS: These results suggest that NDM has the potential to reduce either the proliferation of P. gingivalis, T. forsythia and T. denticola in periodontal pockets or their proteinase-mediated destructive process occurring in periodontitis. PMID- 16473920 TI - Enterobacter gergoviae and the prevalence of efflux in parabens resistance. AB - OBJECTIVES: In order to characterize the mechanism involved in parabens resistance, we studied 13 Enterobacter gergoviae collected from diverse cosmetic formulations containing parabens as preservatives and 10 isolates from clinical or industrial sources. METHODS: RAPD and ERIC-PCR were employed and compared for the epidemiological typing. To study antibiotic and paraben susceptibility, the standard disc diffusion method and the 2-fold dilution method in Luria-Bertani medium were used. Characterization of porins was performed using immunodetection with polyclonal antibodies. Resistance mechanisms against parabens membrane permeabilization were evaluated by measuring K(+) efflux using a specific electrode. mar regulon identification and comparison were carried out. RESULTS: Epidemiological typing confirmed that most of the cosmetic formulations were contaminated by unrelated strains. All of the E. gergoviae strains presented high methylparaben MICs, ranging from 1 to 3.8 g/L, values that were 2-5 times higher than for Escherichia coli or Enterobacter aerogenes, even in strains overexpressing MarA. These MICs decreased in the presence of phenylalanine arginine beta-naphthylamide, pinpointing efflux as a major mechanism of parabens resistance even in E. gergoviae clinical strains. CONCLUSIONS: This is the first report showing the role of active efflux in the parabens resistance in E. gergoviae, a mechanism that may explain its frequent isolation in parabens containing cosmetics compared with other enterobacterial species. Paraben efflux seems to be regulated by a mar-independent process in E. gergoviae. PMID- 16473921 TI - Prevention of urinary tract infection in spinal cord-injured patients: safety and efficacy of a weekly oral cyclic antibiotic (WOCA) programme with a 2 year follow up--an observational prospective study. AB - POPULATION: Spinal cord injury (SCI) patients with neurogenic bladder have an increased risk for symptomatic urinary tract infection (UTI). Recurrent UTI requires multiple courses of antibiotic therapy, markedly increasing the incidence of multidrug-resistant (MDR) bacteria. METHODS: During an observational prospective study, we determined the safety and efficacy of a weekly oral cyclic antibiotic (WOCA) regimen to prevent UTI in SCI adult patients with neurogenic bladder undergoing clean intermittent catheterization. The WOCA regimen consisted of the alternate administration of an antibiotic once per week over a period of at least 2 years. The antibiotics chosen were efficient for UTI, well tolerated and with low selection pressure. RESULTS: There was a significant decrease in antimicrobial consumption linked to the dramatic decrease in the incidence of UTI. Before intervention, there were 9.4 symptomatic UTIs per patient-year, including 197 episodes of febrile UTI responsible for 45 hospitalizations. Under the WOCA regimen there were 1.8 symptomatic UTIs per patient-year, including 19 episodes of febrile UTI. No severe adverse events and no new cases of colonization with MDR bacteria were reported. CONCLUSIONS: In this prospective, observational pilot study a novel approach to the prevention and treatment of UTI in SCI was investigated. Our study shows the benefit of WOCA in preventing UTI in SCI patients. PMID- 16473922 TI - Is personality a determinant of patient satisfaction with hospital care? AB - OBJECTIVE: We investigated to what extent personality is associated with patient satisfaction with hospital care. A sizeable association with personality would render patient satisfaction invalid as an indicator of hospital care quality. DESIGN: Overall satisfaction and satisfaction with aspects of care were regressed on the Big Five dimensions of personality, controlled for patient characteristics as possible explanatory variables of observed associations. PARTICIPANTS: A total of 237 recently discharged inpatients aged 18-84 years (M = 50, SD = 17 years), 57% female, who were hospitalized for an average of 8 days. INSTRUMENTS: The Satisfaction with Hospital Care Questionnaire addressing 12 aspects of care ranging from admission procedures to discharge and aftercare and the Five-Factor Personality Inventory assessing a person's standing on Extraversion, Agreeableness, Conscientiousness, Emotional stability, and Autonomy. RESULTS: Agreeableness significantly predicted patient satisfaction in about half of the scales. After controlling for shared variance with age and educational level, the unique contribution of Agreeableness shrank to a maximum of 3-5% explained variance. When one outlier was dropped from the analysis, the contribution of Agreeableness was no longer statistically significant. CONCLUSION: Patient satisfaction seems only marginally associated with personality, at least at the level of the broad Big Five dimensions. PMID- 16473923 TI - Longitudinal change in food habits between adolescence (11-12 years) and adulthood (32-33 years): the ASH30 Study. AB - BACKGROUND: Links between diet in childhood and the prevention of disease in adulthood have been established. This longitudinal dietary survey provided quantitative evidence of dietary change from adolescence to adulthood, in Northumberland, England. OBJECTIVE: To report longitudinal dietary change in 198 respondents between the ages of 12 and 33 years, to explore dietary 'tracking' between the same time points and to describe the effects of gender, socio economic status and location on dietary change. METHODS: A longitudinal study recorded dietary change from adolescence to adulthood. Two 3-day food diaries were collected in 1980 and 2000 from the same 198 respondents. Foods consumed were assigned to the five categories in the Balance of Good Health (BGH). Demographic and socio-economic information were obtained in 1980 and 2000. RESULTS: Intakes of foods containing fat and/ or sugar and milk and dairy foods decreased (p < 0.01 and p < 0.031, respectively), while intakes of fruits and vegetables increased (p < 0.01). Intakes of bread, other cereals and potatoes (p = 0.002, r = +0.219); fruits and vegetables (p < 0.01, r = +0.256) and meat, fish and alternatives (p = 0.026, r = +0.158) 'tracked' from adolescence to adulthood. Men had increased intake from meat, fish and alternatives and decreased milk and dairy foods more than female respondents (p = 0.003 and p = 0.019). Respondents who had moved away from Northumberland had a greater increase in intake of fruits and vegetables compared with those who remained in the local (p = 0.010). Individuals who had moved to a lower socio-economic group had increased their intake of bread, other cereals and potatoes (p = 0.040). CONCLUSION: Food intake changed considerably in a direction more in the line with current dietary recommendations. Food intake in adolescence was a significant, but not strong, predictor of intake in adulthood. Dietary change is influenced by variables including gender, location and socio-economic status. PMID- 16473924 TI - Retinoic acid can induce markers of endocrine transdifferentiation in pancreatic ductal adenocarcinoma: preliminary observations from an in vitro cell line model. AB - BACKGROUND AND HYPOTHESIS: The pancreatic ductal adenocarcinoma (HPAF) cells have a multipotent stem cell potential. It was hypothesised that all-trans-retinoic acid (atRA) can induce transdifferentiation of these cells into cells with an endocrine phenotype. MATERIAL AND METHODS: To explore this hypothesis, an in vitro system of cells was established. Some cells were treated with atRA at concentrations of 100 nmol/l (non-apoptosis-inducing) and 5 micromol/l (apoptosis inducing) and harvested. Cells were examined for cell cycle kinetics, apoptosis (terminal deoxynucleotidyl transferase assay and p53 protein expression) and immunomorphological features of redifferentiation (MUC1 and DUPAN-2) and endocrine transdifferentiation (insulin, somatostatin, glucagon, neurone-specific enolase) by using immunoperoxidase staining methods. Levels of insulin, transforming growth factor (TGF) beta2, TGFalpha and epidermal growth factor receptor (EGFR) were measured by enzyme-linked immunosorbent assay (ELISA). The vehicle-treated cells served as a control group. RESULTS: When compared with untreated cells, cells treated with 100 nmol/l and 5 micromol/l atRA were observed to show (1) decreased proliferative activity (cpm) as indicated by decreased incorporation of thymidine labelled with hydrogen-3; (2) cell cycle arrest; (3) increased apoptotic activity associated with p53 protein overexpression; (4) upregulated expression of the transdifferentiation and redifferentiation markers; (5) morphological changes indicative of transdifferentiation (increased cell size and appearance of dendrites); (6) decreased production of EGFR; (7) upregulation of TGFalpha and TGFbeta2; and (8) increase in basal and glucose-induced insulin secretion. CONCLUSIONS: Functional endocrine transdifferentiation can be induced in HPAF lines by atRA. Further investigations are mandated to explore the underlying mechanisms of this transdifferentiation and to explore its in vivo extrapolation. PMID- 16473925 TI - The national head and neck histopathology external quality assurance scheme: evolution, design, and analysis of 11 circulations from 1999 to 2005. AB - The background and design of a specialised head and neck histopathology external quality assurance (assessment) scheme with optional oral and ENT arms is described. Participation (submission of response forms) exceeded 85% in eight of 11 slide circulations. In 60% of the 168 circulated cases, up to 33% (and in occasional cases, up to 56%) of respondents gave incomplete or wrong responses owing to a lack of awareness of specific diagnostic entities or to the misinterpretation of specific histological features. In all, there were 118 wrong responses, of which 58% could be considered "potentially serious". The scheme incorporates several design features which optimise the educational value. The stringent marking of cases, and the flagging of performances showing "a meaningful deviation in standard" based on the decision of peer members rather than a rigid numerical formula, encourages high standards. PMID- 16473926 TI - Relationship of p53, Bcl-2, Ki-67 index and E-cadherin expression in early invasive breast cancers with comedonecrosis as an accelerated apoptosis. AB - AIMS: To study the relationship between comedonecrosis formation and morphology, apoptosis, and p53, Bcl-2, Ki-67 index and E-cadherin expression in early invasive breast cancer. EXPERIMENTAL DESIGN: Early invasive breast cancers were first divided into two groups according to the presence (CN+ tumours) or absence (CN- tumours) of comedonecrosis. The histological grade, apoptosis, and expression of E-cadherin, Ki-67, p53 and Bcl-2 in the cancer-affected area, and in normal ducts from the specimen, were then examined. RESULTS: Less tubule and gland formation was seen in CN+ tumours than in CN- tumours, although the histological grade between the groups was not different. During early comedonecrosis, cells undergo apoptosis and subsequent necrosis. p53 was higher in CN+ tumours than in CN- tumours and normal ducts, whereas Bcl-2 was lower in CN+ tumours than in CN- tumours and normal ducts. Both tumours had higher Ki-67 than in normal ducts, but no difference was evident between the tumours. CN+ tumours had slightly higher E-cadherin than that in CN- tumours, but lower than that in normal ducts. The level of comedonecrosis was positively correlated with p53, but inversely correlated with Bcl-2 in all tumours, and p53 and Bcl-2 were inversely correlated with each other. Furthermore, comedonecrosis and p53 were correlated with Ki-67 in CN+ tumours, and Bcl-2 was correlated with Ki-67 in CN- tumours. CONCLUSION: Comedonecrosis may be actively regulated through an apoptotic procedure in massive cancers for their survival and progression, and the above proteins may be associated cooperatively in this process. CN+ and CN- tumours may have opposite proliferative systems under the p53-Bcl-2 pathway. PMID- 16473927 TI - Carriage of group B streptococcus in pregnant women from Oxford, UK. AB - OBJECTIVE: To investigate asymptomatic vagino-rectal carriage of group B streptococcus (GBS) in pregnant women. METHODS: Women in the final trimester of pregnancy were recruited. A single vagino-rectal swab was taken, with consent, for culture of GBS. Two microbiological methods for isolation of GBS from vagino rectal swabs were compared. The distribution of capsular serotypes of the GBS identified was determined. Epidemiological data for a subset (n = 167) of the pregnant women participating were examined. RESULTS: 21.3% were colonised vagino rectally with GBS. Risk factors for neonatal GBS disease (maternal fever, prolonged rupture of membranes, and preterm delivery) were present in 34 of 167 women (20.4%), and the presence of these factors correlated poorly with GBS carriage. Capsular serotypes III (26.4%), IA (25.8%), V (18.9%), and IB (15.7%) were prevalent in the GBS isolates. Selective broth culture of vagino-rectal swabs was superior to selective plate culture, but the combination of both methods was associated with increased detection of GBS (7.5%). An algorithm for the identification of GBS from vagino-rectal swabs was developed. CONCLUSIONS: GBS carriage is prevalent in pregnant women in Oxfordshire, UK. The poor correlation between risk factors and GBS carriage requires further investigation in larger groups, given that the identification of these surrogate markers is recommended to guide administration of intrapartum antibiotic prophylaxis by the Royal College of Obstetricians of the UK. A selective broth culture detected more GBS carriers than a selective plate culture. PMID- 16473929 TI - A comparison of stent-induced stenosis in coronary and peripheral arteries. AB - BACKGROUND AND OBJECTIVES: Restenosis is a complication of interventional procedures such as angioplasty and stenting, often limiting the success of these procedures. Knowledge regarding the relative behaviour of different arteries after these procedures is limited, despite the extensive use of different vascular models. Although the results from studies using different vessels are analysed to predict the behaviour of coronary arteries and other vasculature, direct controlled comparisons between different arteries are necessary for a better understanding of the differential response to restenosis. METHODS: This study examines the response to stenting in coronary and internal iliac arteries as characterised by intimal hyperplasia and restenosis. In a swine model of in stent stenosis, coronary arteries exhibited higher levels of intimal hyperplasia and per cent stenosis than internal iliac arteries. RESULTS: After normalisation for injury score, coronary arteries were found to undergo 47% more intimal hyperplasia (p<0.05), whereas per cent stenosis normalised for injury score tended to be higher (p = 0.01). Other measurements reflecting post-stenting intimal hyperplasia (maximal intimal thickness, medial area) did not exhibit significant differences between the artery groups. CONCLUSIONS: These results show that coronary vessels are more prone to develop significant intimal hyperplasia and subsequent restenosis than internal iliac vessels. A better insight into how different arteries and arterial components behave is important in understanding and developing newer and better therapeutic measures for restenosis. PMID- 16473928 TI - EpCAM and gpA33 are markers of Barrett's metaplasia. AB - AIMS: To characterise a specific and sensitive marker of Barrett's metaplasia (BM). METHODS: Cases of normal oesophageal squamous mucosa (11 fresh endoscopic biopsies and 10 formalin fixed, paraffin embedded tissue blocks), BM (11 biopsies and 11 tissue blocks), and normal gastric body mucosa (five biopsies and five tissue blocks) were analysed using reverse transcriptase PCR, Western blotting, and immunohistochemistry for EpCAM, and reverse transcriptase PCR for gpA33. RESULTS: Strong EpCAM mRNA expression was detected in all the BM cases, in contrast to weak expression in all the normal gastric mucosal samples and no expression in any of the normal oesophageal mucosal samples tested. Strong gpA33 mRNA expression was detected in all the BM cases, in contrast to weak expression in a quarter of the normal gastric mucosal samples and no expression in any of the normal oesophageal mucosal samples tested. Western blotting showed EpCAM protein expression in all the BM cases and in none of the normal gastric or oesophageal mucosal samples tested. Immunohistochemistry showed strong EpCAM protein expression in BM and complete absence of expression in normal oesophageal squamous epithelium. Scattered EpCAM expressing cells were found in the gland bases of normal gastric body mucosa. CONCLUSIONS: EpCAM protein and gpA33 mRNA expressions are specific and sensitive markers of BM. PMID- 16473930 TI - How successful is repeat testicular sperm extraction in patients with azoospermia? AB - BACKGROUND: Little is known about the extraction rate in repeated sperm retrieval procedures in azoospermic patients. This study aimed to assess the feasibility of repeated sperm recovery in these patients. METHODS: A total of 1066 azoospermic men had their first sperm recovery between 1 January 1995 and 31 December 2003. A total of 381 men had obstructive azoospermia (OA), 628 nonobstructive azoospermia (NOA) and 57 showed hypospermatogenesis. RESULTS: Overall, sperm could be retrieved in all procedures in the 598 cycles performed in OA men (100%). A total of 117, 57, 24, 11, 7 and 1 men underwent, respectively, two, three, four, five, six and seven sperm retrievals; all were successful. Of the 784 procedures performed on the 628 men with NOA, sperm could be retrieved in 384 procedures (49%). During the first testicular sperm extraction (TESE) procedure, sperm could be extracted in 261 men with NOA (41.6%). A total of 103 men had a second attempt, 34 had a third attempt, 11 had a fourth attempt, 6 had a fifth attempt and 2 had a sixth attempt. In these cycles, sperm could be extracted in, respectively, 77 (74.7%), 28 (82.3%), 11 (100%), 5 (83.3%) and 2 (100%) men. CONCLUSION: Repeated TESE ensures a high sperm recovery rate even in patients with NOA. In NOA patients, studies reporting on TESE may therefore overestimate the retrieval rate by reallocating successful patients. These data also show that when no spermatozoa can be obtained after thawing cryopreserved testicular sperm for ICSI in NOA patients, a repeat TESE procedure can be planned. PMID- 16473932 TI - Depletion "skraps" and dynamic buffering inside the cellular calcium store. AB - Ca2+ signals, produced by Ca2+ release from cellular stores, switch metabolic responses inside cells. In muscle, Ca2+ sparks locally exhibit the rapid start and termination of the cell-wide signal. By imaging Ca2+ inside the store using shifted excitation and emission ratioing of fluorescence, a surprising observation was made: Depletion during sparks or voltage-induced cell-wide release occurs too late, continuing to progress even after the Ca2+ release channels have closed. This finding indicates that Ca2+ is released from a "proximate" compartment functionally in between store lumen and cytosol. The presence of a proximate compartment also explains a paradoxical surge in intrastore Ca2+, which was recorded upon stimulation of prolonged, cell-wide Ca2+ release. An intrastore surge upon induction of Ca2+ release was first reported in subcellular store fractions, where its source was traced to the store buffer, calsequestrin. The present results update the evolving concept, largely due to N. Ikemoto and C. Kang, of calsequestrin as a dynamic store. Given the strategic location and reduction of dimensionality of Ca2+-adsorbing linear polymers of calsequestrin, they could deliver Ca2+ to the open release channels more efficiently than the luminal store solution, thus constituting the proximate compartment. When store depletion becomes widespread, the polymers would collapse to increase store [Ca2+] and sustain the concentration gradient that drives release flux. PMID- 16473931 TI - Establishment of multiple sublineages of H5N1 influenza virus in Asia: implications for pandemic control. AB - Preparedness for a possible influenza pandemic caused by highly pathogenic avian influenza A subtype H5N1 has become a global priority. The spread of the virus to Europe and continued human infection in Southeast Asia have heightened pandemic concern. It remains unknown from where the pandemic strain may emerge; current attention is directed at Vietnam, Thailand, and, more recently, Indonesia and China. Here, we report that genetically and antigenically distinct sublineages of H5N1 virus have become established in poultry in different geographical regions of Southeast Asia, indicating the long-term endemicity of the virus, and the isolation of H5N1 virus from apparently healthy migratory birds in southern China. Our data show that H5N1 influenza virus, has continued to spread from its established source in southern China to other regions through transport of poultry and bird migration. The identification of regionally distinct sublineages contributes to the understanding of the mechanism for the perpetuation and spread of H5N1, providing information that is directly relevant to control of the source of infection in poultry. It points to the necessity of surveillance that is geographically broader than previously supposed and that includes H5N1 viruses of greater genetic and antigenic diversity. PMID- 16473933 TI - Voltage-gated ion channels in the axon initial segment of human cortical pyramidal cells and their relationship with chandelier cells. AB - The axon initial segment (AIS) of pyramidal cells is a critical region for the generation of action potentials and for the control of pyramidal cell activity. Here we show that Na+ and K+ voltage-gated channels, together with other molecules involved in the localization of ion channels, are distributed asymmetrically in the AIS of pyramidal cells situated in the human temporal neocortex. There is a high density of Na+ channels distributed along the length of the AIS together with the associated proteins spectrin betaIV and ankyrin G. In contrast, Kv1.2 channels are associated with the adhesion molecule Caspr2, and they are mostly localized to the distal region of the AIS. In general, the distal region of the AIS is targeted by the GABAergic axon terminals of chandelier cells, whereas the proximal region is innervated, mostly by other types of GABAergic interneurons. We suggest that this molecular segregation and the consequent regional specialization of the GABAergic input to the AIS of pyramidal cells may have important functional implications for the control of pyramidal cell activity. PMID- 16473934 TI - An unexpected antibody response to an engineered influenza virus modifies CD8+ T cell responses. AB - The ovalbumin(323-339) peptide that binds H2I-A(b) was engineered into the globular heads of hemagglutinin (H) molecules from serologically non-cross reactive H1N1 and H3N2 influenza A viruses, the aim being to analyze recall CD4+ T cell responses in a virus-induced respiratory disease. Prime/challenge experiments with these H1ova and H3ova viruses in H2(b) mice gave the predicted, ovalbumin-specific CD4+ T cell response but showed an unexpectedly enhanced, early expansion of viral epitope-specific CD8+ T cells in spleen and a greatly diminished inflammatory process in the virus-infected respiratory tract. At the same time, the primary antibody response to the H3N2 challenge virus was significantly reduced, an effect that has been associated with preexisting neutralizing antibody in other experimental systems. Analysis of serum from the H1ova-primed mice showed low-level binding to H3ova but not to the wild-type H3N2 virus. Experiments with CD4+ T cell-depleted and Ig-/- mice indicated that this cross-reactive Ig is indeed responsible for the modified pathogenesis after respiratory challenge. Furthermore, the effect does not seem to be virus-dose related, although it does require infection. These findings suggest intriguing possibilities for vaccination and, at the same time, emphasize that engineered modifications in viruses may have unintended immunological consequences. PMID- 16473936 TI - Partial tuning of motor cortex neurons to final posture in a free-moving paradigm. AB - Motor cortex neurons in the monkey brain were tested with a diverse and naturalistic arm movement set. Over this global set of movements, the neurons showed a limited but significant degree of tuning to the multijoint posture attained by the arm at the end of each movement. Further supporting the hypothesis that the neurons are partially tuned to end posture, the postures preferred by the neurons significantly matched the postures evoked by electrical stimulation of the same cortical sites. However, much of the variance in neuronal activity remained unexplained even by the end-posture model, and thus other variables must have contributed to the response profile of the neurons. One possibility is that motor cortex neurons become tuned to the wide variety of movement parameters that are relevant to the animal's normal behavioral repertoire, and, therefore, any one parameter accounts for only a limited amount of neuronal variance. PMID- 16473935 TI - The DDB1-CUL4ADDB2 ubiquitin ligase is deficient in xeroderma pigmentosum group E and targets histone H2A at UV-damaged DNA sites. AB - Xeroderma pigmentosum (XP) is a heritable human disorder characterized by defects in nucleotide excision repair (NER) and the development of skin cancer. Cells from XP group E (XP-E) patients have a defect in the UV-damaged DNA-binding protein complex (UV-DDB), involved in the damage recognition step of NER. UV-DDB comprises two subunits, products of the DDB1 and DDB2 genes, respectively. Mutations in the DDB2 gene account for the underlying defect in XP-E. The UV-DDB complex is a component of the newly identified cullin 4A-based ubiquitin E3 ligase, DDB1-CUL4A(DDB2). The E3 ubiquitin ligases recognize specific substrates and mediate their ubiquitination to regulate protein activity or target proteins for degradation by the proteasomal pathway. In this study, we have addressed the role of the UV-DDB-based E3 in NER and sought a physiological substrate. We demonstrate that monoubiquitinated histone H2A in native chromatin coimmunoprecipitates with the endogenous DDB1-CUL4A(DDB2) complex in response to UV irradiation. Further, mutations in DDB2 alter the formation and binding activity of the DDB1-CUL4A(DDB2) ligase, accompanied by impaired monoubiquitination of H2A after UV treatment of XP-E cells, compared with repair proficient cells. This finding indicates that DDB2, as the substrate receptor of the DDB1-CUL4A-based ligase, specifically targets histone H2A for monoubiquitination in a photolesion-binding-dependent manner. Given that the loss of monoubiquitinated histone H2A at the sites of UV-damaged DNA is associated with decreased global genome repair in XP-E cells, this study suggests that histone modification, mediated by the XPE factor, facilitates the initiation of NER. PMID- 16473937 TI - Z-DNA-forming sequences generate large-scale deletions in mammalian cells. AB - Spontaneous chromosomal breakages frequently occur at genomic hot spots in the absence of DNA damage and can result in translocation-related human disease. Chromosomal breakpoints are often mapped near purine-pyrimidine Z-DNA-forming sequences in human tumors. However, it is not known whether Z-DNA plays a role in the generation of these chromosomal breakages. Here, we show that Z-DNA-forming sequences induce high levels of genetic instability in both bacterial and mammalian cells. In mammalian cells, the Z-DNA-forming sequences induce double strand breaks nearby, resulting in large-scale deletions in 95% of the mutants. These Z-DNA-induced double-strand breaks in mammalian cells are not confined to a specific sequence but rather are dispersed over a 400-bp region, consistent with chromosomal breakpoints in human diseases. This observation is in contrast to the mutations generated in Escherichia coli that are predominantly small deletions within the repeats. We found that the frequency of small deletions is increased by replication in mammalian cell extracts. Surprisingly, the large-scale deletions generated in mammalian cells are, at least in part, replication independent and are likely initiated by repair processing cleavages surrounding the Z-DNA-forming sequence. These results reveal that mammalian cells process Z DNA-forming sequences in a strikingly different fashion from that used by bacteria. Our data suggest that Z-DNA-forming sequences may be causative factors for gene translocations found in leukemias and lymphomas and that certain cellular conditions such as active transcription may increase the risk of Z-DNA related genetic instability. PMID- 16473938 TI - Effects of cyclothiazide on GluR1/AMPA receptors. AB - Cyclothiazide (CTZ), a positive allosteric modulator of ionotropic alpha-amino-3 hydroxy-5-methylisoxazole-4-propionic acid (AMPA)-type glutamate receptors, is used frequently to block the desensitization of both native and heterologously expressed AMPA receptors. Specifically, CTZ is known to produce a fast inhibition of AMPA receptor desensitization and a much slower potentiation of the AMPA current. By using patch-clamp techniques, the effects of CTZ were studied in HEK 293 cells stably transfected with the rat flip GluR1 subunit. Upon CTZ treatment, we found an increased apparent affinity for the agonist, a slow whole-cell current potentiation, a fast inhibition of desensitization, and a lengthening of single-channel openings. Furthermore, we show that CTZ alters the channel gating events modifying the relative contribution of different single-channel classes of conductance (gamma), increasing and decreasing, respectively, the contributions of gammaM (medium) and gammaL (low) without altering that of the gammaH (high) conductance channels. We also present a kinetic model that predicts well all of the experimental findings of CTZ action. Finally, we suggest a protocol for standard cell treatment with CTZ to attain maximal efficacy of CTZ on GluR1 receptors. PMID- 16473939 TI - Inhibition of ERK pathway or protein synthesis during reexposure to drugs of abuse erases previously learned place preference. AB - Repeated association of drugs of abuse with context leads to long-lasting behavioral responses that reflect reward-controlled learning and participate in the establishment of addiction. Reactivation of consolidated memories is known to produce a reconsolidation process during which memories undergo a labile state. We investigated whether reexposure to drugs had similar effects. Cocaine administration activates extracellular signal-regulated kinase (ERK) in the striatum, and ERK activation is required for the acquisition of cocaine-induced conditioned place preference (CPP). When mice previously conditioned for cocaine place preference were reexposed to cocaine in the drug-paired compartment after systemic administration of SL327, an inhibitor of ERK activation, CPP response was abolished 24 h later. This procedure also abolished the phosphorylation of ERK and glutamate receptor-1 observed in the ventral and dorsal striatum, 24 h later, during CPP test. Erasure of CPP by SL327 required the combination of cocaine administration and drug-paired context and did not result from enhanced extinction. Similarly, reexposure to morphine in the presence of SL327 long lastingly abolished response of previously learned morphine-CPP. The effects of SL327 on cocaine- or morphine-CPP were reproduced by protein synthesis inhibition. In contrast, protein synthesis inhibition did not alter previously acquired locomotor sensitization to cocaine. Our findings show that an established CPP can be disrupted when reactivation associates both the conditioned context and drug administration. This process involves ERK, and systemic treatment preventing ERK activation during reexposure erases the previously learned behavioral response. These results suggest potential therapeutic strategies to explore in the context of addiction. PMID- 16473941 TI - Patterned variation in prehistoric chiefdoms. AB - Comparative study of early complex societies (chiefdoms) conjures visions of a cultural evolutionary emphasis on similarities and societal typology. Variation within the group has not been as systematically examined but offers an even more productive avenue of approach to fundamental principles of organization and change. Three widely separated trajectories of early chiefdom development are compared here: the Valley of Oaxaca (Mexico), the Alto Magdalena (Colombia), and Northeast China. Archaeological data from all three regions are analyzed with the same tools to reveal variation in human activities, relationships, and interactions as these change in the emergence of chiefly communities. Patterning in this variation suggests the operation of underlying general principles, which are offered as hypotheses that merit further investigation and evaluation in comparative study of a much larger number of cases. PMID- 16473940 TI - Frequency selectivity of synaptic exocytosis in frog saccular hair cells. AB - The ability to respond selectively to particular frequency components of sensory inputs is fundamental to signal processing in the ear. The frog (Rana pipiens) sacculus, which is used for social communication and escape behaviors, is an exquisitely sensitive detector of sounds and ground-borne vibrations in the 5- to 200-Hz range, with most afferent axons having best frequencies between 40 and 60 Hz. We monitored the synaptic output of saccular sensory receptors (hair cells) by measuring the increase in membrane capacitance (deltaC(m)) that occurs when synaptic vesicles fuse with the plasmalemma. Strong stepwise depolarization evoked an exocytic burst that lasted 10 ms and corresponded to the predicted capacitance of all docked vesicles at synapses, followed by a 20-ms delay before additional vesicle fusion. Experiments using weak stimuli, within the normal physiological range for these cells, revealed a sensitivity to the temporal pattern of membrane potential changes. Interrupting a weak depolarization with a properly timed hyperpolarization increased deltaC(m). Small sinusoidal voltage oscillations (+/-5 mV centered at -60 mV) evoked a deltaC(m) that corresponded to 95 vesicles per s at each synapse at 50 Hz but only 26 vesicles per s at 5 Hz and 27 vesicles per s at 200 Hz (perforated patch recordings). This frequency selectivity was absent for larger sinusoidal oscillations (+/-10 mV centered at 55 mV) and was largest for hair cells with the smallest sinusoidal-stimuli-evoked Ca2+ currents. We conclude that frog saccular hair cells possess an intrinsic synaptic frequency selectivity that is saturated by strong stimuli. PMID- 16473942 TI - Conformal invariants associated to a measure. AB - In this note, we study some conformal invariants of a Riemannian manifold (M(n), g) equipped with a smooth measure m. In particular, we show that there is a natural definition of the Ricci and scalar curvatures associated to such a space, both of which are conformally invariant. We also adapt the methods of Fefferman and Graham [Fefferman, C. & Graham, C. R. (1985) Asterisque, Numero Hors Serie, 95-116] and Graham, Jenne, Mason, and Sparling [Graham, C. R., Jenne, R., Mason, L. J., & Sparling, G. A. J. (1992) J. London Math. Soc. 46, 557-565] to construct families of conformally covariant operators defined on these spaces. Certain variational problems in this setting are considered, including a generalization of the Einstein-Hilbert action. PMID- 16473943 TI - Cell-type-selective induction of c-jun by TAF4b directs ovarian-specific transcription networks. AB - Cell-type-selective expression of the TFIID subunit TAF(II)105 (renamed TAF4b) in the ovary is essential for proper follicle development. Although a multitude of signaling pathways required for folliculogenesis have been identified, downstream transcriptional integrators of these signals remain largely unknown. Here, we show that TAF4b controls the granulosa-cell-specific expression of the proto oncogene c-jun, and together they regulate transcription of ovary-selective promoters. Instead of using cell-type-specific activators, our findings suggest that the coactivator TAF4b regulates the expression of tissue-specific genes, at least in part, through the cell-type-specific induction of c-jun, a ubiquitous activator. Importantly, the loss of TAF4b in ovarian granulosa cells disrupts cellular morphologies and interactions during follicle growth that likely contribute to the infertility observed in TAF4b-null female mice. These data highlight a mechanism for potentiating tissue-selective functions of the basal transcription machinery and reveal intricate networks of gene expression that orchestrate ovarian-specific functions and cell morphology. PMID- 16473944 TI - Effective expansion of alloantigen-specific Foxp3+ CD25+ CD4+ regulatory T cells by dendritic cells during the mixed leukocyte reaction. AB - Thymic-derived CD25+ CD4+ T regulatory cells (Tregs) suppress immune responses, including transplantation. Here we evaluated the ability of dendritic cells (DCs) to expand alloantigen-specific Tregs in the mixed leukocyte reaction (MLR) that develops from polyclonal populations of T cells. The allogeneic DCs, when supplemented with IL-2 in the cultures, were much more effective than bulk spleen cells in expanding the numbers of Tregs. Likewise, DCs and not spleen cells were effective in sustaining expression of the transcription factor Foxp3 in Tregs, but neither IL-2 nor CD80/86 was required for this effect in the cultures. On a per-cell basis, the DC-expanded, but not unexpanded, Tregs were more potent suppressors of a fresh MLR by CD25- CD4+ T cells. Suppression was 3- to 10-fold more active for MLRs induced by the original alloantigens than for third-party stimulators. When DC-expanded Tregs were introduced into sublethally irradiated hosts, the T cells suppressed graft-versus-host-disease induced by CD25- CD4+ T cells. Again, suppression was more active against the same mouse strain that provided the DCs to expand the Tregs. Therefore, alloantigen-selected Tregs are more effective suppressors of responses to major transplantation antigens, and these Tregs can be expanded from a polyclonal repertoire by DCs. PMID- 16473945 TI - Geography and macroeconomics: new data and new findings. AB - The linkage between economic activity and geography is obvious: Populations cluster mainly on coasts and rarely on ice sheets. Past studies of the relationships between economic activity and geography have been hampered by limited spatial data on economic activity. The present study introduces data on global economic activity, the G-Econ database, which measures economic activity for all large countries, measured at a 1 degree latitude by 1 degree longitude scale. The methodologies for the study are described. Three applications of the data are investigated. First, the puzzling "climate-output reversal" is detected, whereby the relationship between temperature and output is negative when measured on a per capita basis and strongly positive on a per area basis. Second, the database allows better resolution of the impact of geographic attributes on African poverty, finding geography is an important source of income differences relative to high-income regions. Finally, we use the G-Econ data to provide estimates of the economic impact of greenhouse warming, with larger estimates of warming damages than past studies. PMID- 16473947 TI - Testicular germ cells can colonize sexually undifferentiated embryonic gonad and produce functional eggs in fish. AB - Understanding the mechanisms that regulate germ-cell development is crucial to reproductive medicine and animal production. Animal gametes originally derive from sexually undifferentiated primordial germ cells (PGCs), which develop into mitotic germ cells (oogonia or spermatogonia) before proceeding to meiosis [Wylie, C. (1999) Cell 96, 165-174]. Spermatogonia are thought to include a population of cells with stem cell activity, which proliferate throughout the lifespan of male animals and produce spermatozoa [Zhao, G. Q. & Garbers, D. L. (2002) Dev. Cell 2, 537-547]. However, the functional differences between PGCs and spermatogonial stem cells are poorly understood. Here we show that transplanted adult testicular germ cells can colonize sexually undifferentiated embryonic gonads and resume gametogenesis. Testicular germ cells containing spermatogonial stem cells isolated from adult male rainbow trout (Oncorhynchus mykiss) were transplanted into the peritoneal cavity of newly hatched embryos of both sexes, and the behavior of the donor cells was observed. The testicular germ cells differentiated into spermatozoa in male recipients and fully functional eggs in female recipients. Furthermore, the donor-derived spermatozoa and eggs obtained from the recipient fish were able to produce normal offspring. These findings indicate that fish testicular germ cells, probably spermatogonial stem cells, possess a high level of developmental plasticity and sexual bipotency, even after the animal reaches maturity. Furthermore, our results suggest that spermatogonial stem cells are at least partly functionally similar to PGCs. PMID- 16473946 TI - Regulation of antibacterial defense in the small intestine by the nuclear bile acid receptor. AB - Obstruction of bile flow results in bacterial proliferation and mucosal injury in the small intestine that can lead to the translocation of bacteria across the epithelial barrier and systemic infection. These adverse effects of biliary obstruction can be inhibited by administration of bile acids. Here we show that the farnesoid X receptor (FXR), a nuclear receptor for bile acids, induces genes involved in enteroprotection and inhibits bacterial overgrowth and mucosal injury in ileum caused by bile duct ligation. Mice lacking FXR have increased ileal levels of bacteria and a compromised epithelial barrier. These findings reveal a central role for FXR in protecting the distal small intestine from bacterial invasion and suggest that FXR agonists may prevent epithelial deterioration and bacterial translocation in patients with impaired bile flow. PMID- 16473948 TI - Uncoupling of the base excision and nucleotide incision repair pathways reveals their respective biological roles. AB - The multifunctional DNA repair enzymes apurinic/apyrimidinic (AP) endonucleases cleave DNA at AP sites and 3'-blocking moieties generated by DNA glycosylases in the base excision repair pathway. Alternatively, in the nucleotide incision repair (NIR) pathway, the same AP endonucleases incise DNA 5' of a number of oxidatively damaged bases. At present, the physiological relevance of latter function remains unclear. Here, we report genetic dissection of AP endonuclease functions in base excision repair and NIR pathways. Three mutants of Escherichia coli endonuclease IV (Nfo), carrying amino acid substitutions H69A, H109A, and G149D have been isolated. All mutants were proficient in the AP endonuclease and 3'-repair diesterase activities but deficient in the NIR. Analysis of metal content reveals that all three mutant proteins have lost one of their intrinsic zinc atoms. Expression of the nfo mutants in a repair-deficient strain of E. coli complemented its hypersensitivity to alkylation but not to oxidative DNA damage. The differential drug sensitivity of the mutants suggests that the NIR pathway removes lethal DNA lesions generated by oxidizing agents. To address the physiological relevance of the NIR pathway in human cells, we used the fluorescence quenching mechanism of molecular beacons. We show that in living cells a major human AP endonuclease, Ape1, incises DNA containing alpha-anomeric 2'-deoxyadenosine, indicating that the intracellular environment supports NIR activity. Our data establish that NIR is a distinct and separable function of AP endonucleases essential for handling lethal oxidative DNA lesions. PMID- 16473949 TI - Conserved thermodynamic contributions of backbone hydrogen bonds in a protein fold. AB - Backbone-backbone hydrogen-bonding interactions are a ubiquitous and highly conserved structural feature of proteins that adopt the same fold (i.e., have the same overall backbone topology). This work addresses the question of whether or not this structural conservation is also reflected as a thermodynamic conservation. Reported here is a comparative thermodynamic analysis of backbone hydrogen bonds in two proteins that adopt the same fold but are unrelated at the primary amino acid sequence level. With amide-to-ester bond mutations introduced by total chemical synthesis methods, the thermodynamic consequences of backbone backbone hydrogen-bond deletions at five different structurally equivalent positions throughout the beta-alpha-alpha fold of Arc repressor and CopG were assessed. The ester bond-containing analogues all folded into native-like three dimensional structures that were destabilized from 2.5 to 6.0 kcal/(mol dimer) compared with wild-type controls. Remarkably, the five paired analogues with amide-to-ester bond mutations at structurally equivalent positions were destabilized to exactly the same degree, regardless of the degree to which the mutation site was buried in the structure. The results are interpreted as evidence that the thermodynamics of backbone-backbone hydrogen-bonding interactions in a protein fold are conserved. PMID- 16473951 TI - A macroporous hydrogel for the coculture of neural progenitor and endothelial cells to form functional vascular networks in vivo. AB - A microvascular network is critical for the survival and function of most tissues. We have investigated the potential of neural progenitor cells to augment the formation and stabilization of microvascular networks in a previously uncharacterized three-dimensional macroporous hydrogel and the ability of this engineered system to develop a functional microcirculation in vivo. The hydrogel is synthesized by cross-linking polyethylene glycol with polylysine around a salt leached polylactic-co-glycolic acid scaffold that is degraded in a sodium hydroxide solution. An open macroporous network is formed that supports the efficient formation of tubular structures by brain endothelial cells. After subcutaneous implantation of hydrogel cocultures in mice, blood flow in new microvessels was apparent at 2 weeks with perfused networks established on the surface of implants at 6 weeks. Compared to endothelial cells cultured alone, cocultures of endothelial cells and neural progenitor cells had a significantly greater density of tubular structures positive for platelet endothelial cell adhesion molecule-1 at the 6-week time point. In implant cross sections, the presence of red blood cells in vessel lumens confirmed a functional microcirculation. These findings indicate that neural progenitor cells promote the formation of endothelial cell tubes in coculture and the development of a functional microcirculation in vivo. We demonstrate a previously undescribed strategy for creating stable microvascular networks to support engineered tissues of desired parenchymal cell origin. PMID- 16473950 TI - Maternal mediation, stress inoculation, and the development of neuroendocrine stress resistance in primates. AB - The stress inoculation hypothesis presupposes that brief intermittent stress exposure early in life induces the development of subsequent stress resistance in human and nonhuman primates. Rodent studies, however, suggest a role for maternal care rather than stress exposure per se (i.e., the maternal mediation hypothesis). To investigate these two hypotheses, we examined maternal care and the development of stress resistance after exposure to brief intermittent infant stress (IS), mother-infant stress (MIS), or no stress (NS) protocols administered to 30 monkeys between postnatal weeks 17 and 27. Unlike rodents, the IS condition did not permanently increase primate maternal care, nor did measures of total maternal care predict subsequent offspring hypothalamic-pituitary-adrenal-axis responsivity. Although MIS infants received less maternal care than IS and NS infants, both IS and MIS monkeys developed subsequent stress resistance. These findings indicate that rearing differences in the development of stress resistance are more closely related to differences in prior stress exposure than to differences in maternal care. A second experiment confirmed this conclusion in a different cohort of 25 monkeys exposed as infants to high foraging-demand (HFD) or low foraging-demand (LFD) conditions. HFD infants exhibited intermittent elevations in cortisol levels and received less maternal care than LFD infants. In keeping with a key prediction of the stress inoculation hypothesis, HFD males responded to stress in adulthood with diminished hypothalamic-pituitary-adrenal axis activation compared with LFD males. Results from both experiments demonstrate that stress inoculation, rather than high levels of maternal care, promotes the development of primate stress resistance. PMID- 16473952 TI - Posterior alpha activity is not phase-reset by visual stimuli. AB - There is currently a debate as to whether event-related potentials and fields measured by using electroencephalography or magnetoencephalography are generated by ongoing oscillatory activity becoming phase-reset in response to a given stimulus. We performed a magnetoencephalography study measuring brain activity in response to visual stimuli. Using a measure termed the phase-preservation index we investigated the phase of oscillatory alpha activity (8-13 Hz) before and after the stimulus. We found that in single trials the alpha oscillations after visual stimuli preserve their phase relationship with respect to the phase before the stimuli. This finding argues against phase-resetting of ongoing oscillations as being responsible for visually evoked responses. The event-related field can be explained primarily by stimulus-locked activity in the band that is absent before the stimulus. These findings suggest that different neuronal events are responsible for generating the ongoing oscillations and the visually evoked responses. PMID- 16473953 TI - A study of indoor 220Rn and 222Rn decay product concentrations in the UK. AB - In order to gain a better understanding of the risk of human exposure to 220Rn indoors, measurements of 220Rn decay products have been performed in a number of houses in England. The study focused mainly on areas where above-average indoor 220Rn concentrations were to be expected because of geological or other factors. Thoron (220Rn) in room air comes mainly from the building materials, with an additional contribution from soil gas; therefore, 220Rn concentrations were examined in relation to building materials as well as location. Measurements were carried out in 23 houses. The mean equilibrium equivalent 220Rn (EET) concentration found was 0.39 Bq m-3 and the mean equilibrium equivalent 222Rn (EER) concentration was 17.8 Bq m-3. The 220Rn concentration values and EET/EER ratios found in this investigation correspond well with other published results. Values imply that the 220Rn concentrations in English dwellings are a far less important problem than 222Rn concentrations. PMID- 16473954 TI - Regional heterogeneity in acetylcholine-induced relaxation in rat vascular bed: role of calcium-activated K+ channels. AB - Ca+ -activated K+ -channels (KCa) regulate vasomotor tone via smooth muscle hyperpolarization and relaxation. The relative contribution of the endothelium derived hyperpolarizing factor (EDHF)-mediated relaxation differs depending on vessel type and size. It is unknown whether these KCa channels are differentially distributed along the same vascular bed and hence have different roles in mediating the EDHF response. We therefore assessed the role of small- (SKCa), intermediate- (IKCa), and large-conductance (BKCa) channels in mediating acetylcholine-induced relaxations in both first- and fourth-order side branches of the rat superior mesenteric artery (MA1 and MA4, respectively). Two-millimeter segments of each MA were mounted in the wire myograph, incubated with Nomega nitro-L-arginine methyl ester (L-NAME, 100 micromol/l) and indomethacin (10 micromol/l), and precontracted with phenylephrine (10 micromol/l). Cumulative concentration-response curves to ACh (0.001-10 micromol/l) were performed in the absence or presence of selective KCa channel antagonists. Apamin almost completely abolished these relaxations in MA4 but only partially blocked relaxations in MA1. The selective IKCa channel blocker 1-[(2-chlorophenyl) diphenylmethyl]-1H-pyrazole (TRAM-34) caused a significantly greater inhibition of the ACh-induced relaxation in MA4 compared with MA1. Iberiotoxin had no inhibitory effect in MA4 but blunted relaxation in MA1. Relative mRNA expression levels of SKCa (rSK1, rSK3, and rSK4 = rIK1) were significantly higher in MA4 compared with MA1. BKCa (rBKalpha1 and rBKbeta1) genes were similar in both MA1 and MA4. Our data demonstrate regional heterogeneity in SKCa and IKCa function and gene expression and stress the importance of these channels in smaller resistance-sized arteries, where the role of EDHF is more pronounced. PMID- 16473955 TI - Sex differences in myocardial infarct size are abolished by sarcolemmal KATP channel blockade in rat. AB - This study was conducted to examine the relationship between myocardial ATP sensitive potassium (K(ATP)) channels and sex differences in myocardial infarct size after in vitro ischemia-reperfusion (I/R). Hearts from adult male and female Sprague-Dawley rats were excised and exposed to an I/R protocol (1 h of ischemia, followed by 2 h of reperfusion) on a modified Langendorff apparatus. Hearts from female rats showed significantly smaller infarct sizes than hearts from males (23 +/- 4 vs. 40 +/- 5% of the zone at risk, respectively; P < 0.05). Administration of HMR-1098, a sarcolemmal K(ATP) channel blocker, abolished the sex difference in infarct size (42 +/- 4 vs. 45 +/- 5% of the zone at risk in hearts from female and male rats, respectively; P = not significant). Further experiments showed that blocking the K(ATP) channels in ischemia, and not reperfusion, was sufficient to increase infarct size in female rats. These data demonstrate that sarcolemmal K(ATP) channels are centrally involved in mechanisms that underlie sex differences in the susceptibility of the intact heart to I/R injury. PMID- 16473956 TI - Inhibition of cardiac contractility by 5-hydroxydecanoate and tetraphenylphosphonium ion: a possible role of mitoKATP in response to inotropic stress. AB - This study investigates the role of the mitochondrial ATP-sensitive K+ channel (mitoKATP) in response to positive inotropic stress. In Langendorff-perfused rat hearts, inotropy was induced by increasing perfusate calcium to 4 mM, by adding 80 microM ouabain or 0.25 microM dobutamine. Each of these treatments resulted in a sustained increase in rate-pressure product (RPP) of approximately 60%. Inhibition of mitoKATP by perfusion of 5-hydroxydecanoate (5-HD) or tetraphenylphosphonium before induction of inotropic stress resulted in a marked attenuation of RPP. Inhibition of mitoKATP after induction of stress caused the inability of the heart to maintain a high-work state. In human atrial fibers, the increase in contractility induced by dobutamine was inhibited 60% by 5-HD. In permeabilized fibers from the Langendorff-perfused rat hearts, inhibition of mitoKATP resulted, in all cases, in an alteration of adenine nucleotide compartmentation, as reflected by a 60% decrease in the half-saturation constant for ADP [K1/2 (ADP)]. We conclude that opening of cardiac mitoKATP is essential for an appropriate response to positive inotropic stress and propose that its involvement proceeds through the prevention of stress-induced decrease in mitochondrial matrix volume. These results indicate a physiological role for mitoKATP in inotropy and, by extension, in heart failure. PMID- 16473957 TI - Cardiac fibrogenesis in magnesium deficiency: a role for circulating angiotensin II and aldosterone. AB - Mechanisms underlying cardiac fibrogenesis in magnesium deficiency are unclear. It was reported earlier from this laboratory that serum from magnesium-deficient rats has a more pronounced stimulatory effect on cell proliferation, net collagen production, and superoxide generation in adult rat cardiac fibroblasts than serum from rats on the control diet. The profibrotic serum factors were, however, not identified. This study tested the hypothesis that circulating angiotensin II may modulate cardiac fibroblast activity in hypomagnesemic rats. Male Sprague-Dawley rats were pair-fed a magnesium-deficient (0.0008% Mg) or -sufficient (0.05%) diet for 6 days, and the effects of serum from these rats on [3H]thymidine and [3H]proline incorporation into cardiac fibroblasts from young adult rats were evaluated in the presence of losartan, an angiotensin II type 1 (AT1) receptor antagonist, and spironolactone, an aldosterone antagonist. Losartan and spironolactone markedly attenuated the stimulatory effects in vitro of serum from the magnesium-deficient and control groups, but the inhibitory effects were considerably higher in cells exposed to serum from magnesium-deficient animals. Circulating and cardiac tissue levels of angiotensin II were significantly elevated in magnesium-deficient animals (67.6% and 93.1%, respectively, vs. control). Plasma renin activity was 61.9% higher in magnesium-deficient rats, but serum angiotensin-converting enzyme activity was comparable in the two groups. Furthermore, preliminary experiments in vivo using enalapril supported a role for angiotensin II in magnesium deficiency. There was no significant difference between the groups in serum aldosterone levels. The findings suggest that circulating angiotensin II and aldosterone may stimulate fibroblast activity and contribute to a fibrogenic response in the heart in magnesium deficiency. PMID- 16473958 TI - Production of NAADP and its role in Ca2+ mobilization associated with lysosomes in coronary arterial myocytes. AB - The present study was designed to determine the production of nicotinic acid adenine dinucleotide phosphate (NAADP) and its role associated with lysosomes in mediating endothelin-1 (ET-1)-induced vasoconstriction in coronary arteries. HPLC assay showed that NAADP was produced in coronary arterial smooth muscle cells (CASMCs) via endogenous ADP-ribosyl cyclase. Fluorescence microscopic analysis of intracellular Ca2+ concentration ([Ca2+]i) in CASMCs revealed that exogenous 100 nM NAADP increased [Ca2+]i by 711 +/- 47 nM. Lipid bilayer experiments, however, demonstrated that NAADP did not directly activate ryanodine (Rya) receptor Ca2+ release channels on the sarcoplasmic reticulum. In CASMCs pretreated with 100 nM bafilomycin A1 (Baf), an inhibitor of lysosomal Ca2+ release and vacuolar proton pump function, NAADP-induced [Ca2+]i increase was significantly abolished. Moreover, ET-1 significantly increased NAADP formation in CASMCs and resulted in the rise of [Ca2+]i in these cells with a large increase in global Ca2+ level of 1,815 +/- 84 nM. Interestingly, before this large Ca2+ increase, a small Ca2+ spike with an increase in [Ca2+]i of 529 +/- 32 nM was observed. In the presence of Baf (100 nM), this ET-1-induced two-phase [Ca2+]i response was completely abolished, whereas Rya (50 microM) only markedly blocked the ET-1-induced large global Ca2+ increase. Functional studies showed that 100 nM Baf significantly attenuated ET-1-induced maximal constriction from 82.26 +/- 4.42% to 51.80 +/- 4.36%. Our results suggest that a lysosome-mediated Ca2+ regulatory mechanism via NAADP contributes to ET-1-induced Ca2+ mobilization in CASMCs and consequent vasoconstriction of coronary arteries. PMID- 16473959 TI - Mesenchymal stem cell injection after myocardial infarction improves myocardial compliance. AB - Cellular therapy for myocardial injury has improved ventricular function in both animal and clinical studies, though the mechanism of benefit is unclear. This study was undertaken to examine the effects of cellular injection after infarction on myocardial elasticity. Coronary artery ligation of Lewis rats was followed by direct injection of human mesenchymal stem cells (MSCs) into the acutely ischemic myocardium. Two weeks postinfarct, myocardial elasticity was mapped by atomic force microscopy. MSC-injected hearts near the infarct region were twofold stiffer than myocardium from noninfarcted animals but softer than myocardium from vehicle-treated infarcted animals. After 8 wk, the following variables were evaluated: MSC engraftment and left ventricular geometry by histological methods, cardiac function with a pressure-volume conductance catheter, myocardial fibrosis by Masson Trichrome staining, vascularity by immunohistochemistry, and apoptosis by TdT-mediated dUTP nick-end labeling assay. The human cells engrafted and expressed a cardiomyocyte protein but stopped short of full differentiation and did not stimulate significant angiogenesis. MSC injected hearts showed significantly less fibrosis than controls, as well as less left ventricular dilation, reduced apoptosis, increased myocardial thickness, and preservation of systolic and diastolic cardiac function. In summary, MSC injection after myocardial infarction did not regenerate contracting cardiomyocytes but reduced the stiffness of the subsequent scar and attenuated postinfarction remodeling, preserving some cardiac function. Improving scarred heart muscle compliance could be a functional benefit of cellular cardiomyoplasty. PMID- 16473960 TI - Longitudinal movements and resulting shear strain of the arterial wall. AB - There has been little interest in the longitudinal movement of the arterial wall. It has been assumed that this movement is negligible compared with the diameter change. Using a new high-resolution noninvasive ultrasonic method, we measured longitudinal movements and diameter change of the common carotid artery of 10 healthy humans. During the cardiac cycle, a distinct bidirectional longitudinal movement of the intima-media complex could be observed in all the subjects. An antegrade longitudinal movement, i.e., in the direction of blood flow, in early systole [0.39 mm (SD 0.26)] was followed by a retrograde longitudinal movement, i.e., in the direction opposite blood flow [-0.52 mm (SD 0.27)], later in systole and a second antegrade longitudinal movement [0.41 mm (SD 0.33)] in diastole. The corresponding diameter change was 0.65 mm (SD 0.19). The adventitial region showed the same basic pattern of longitudinal movement; however, the magnitude of the movements was smaller than that of the intima-media complex, thereby introducing shear strain and, thus, shear stress within the wall [maximum shear strain between the intima-media complex and the adventitial region was 0.36 rad (SD 0.26). These phenomena have not previously been described. Measurements were also performed on the abdominal aorta (n = 3) and brachial (n = 3) and popliteal (n = 3) arteries. Our new information seems to be of fundamental importance for further study and evaluation of vascular biology and hemodynamics and, thus, for study of atherosclerosis and vascular diseases. PMID- 16473961 TI - Docosahexaenoic acid induces ciap1 mRNA and protects human endothelial cells from stress-induced apoptosis. AB - Induction of apoptosis represents a potential reaction of endothelial cells (ECs) after injury of the vascular endothelium. Beneficial effects of n-3 polyunsaturated fatty acids (PUFAs) in vascular diseases are widely recognized although the responsible mechanisms are not fully understood. Because it is not known whether PUFAs modulate EC apoptosis, we investigated the effects of n-3 and n-6 PUFAs on 4-hydroxynonenal (HNE)-induced EC apoptosis by annexin V staining and caspase-3 activation assays. Pretreatment with the n-3 fatty acid docosahexaenoic acid (DHA) reduced HNE-induced EC apoptosis. DHA-treated cells did not show the pronounced drop in intracellular GSH after HNE exposure seen in vehicle- or n-6 arachidonic acid-treated cells. This is most likely due to increased GSH levels in DHA-treated cells. Furthermore, DHA pretreatment increased ciap1 mRNA levels and transfection of cIAP1 small interfering RNA abolished the protective effect of DHA in HNE-induced apoptosis in HUVECs. Thus pretreatment of HUVECs with DHA reduces HNE-induced oxidative stress and apoptosis, and the protective effects of DHA seem to be dependent on cIAP1. The results provide a possible new mechanism for the atheroprotective effects of n-3 fatty acids in vascular disease. PMID- 16473962 TI - Alterations in structure and mechanics of resistance arteries from ouabain induced hypertensive rats. AB - We have previously described that chronic administration of ouabain induces hypertension and functional alterations in mesenteric resistance arteries. The aim of this study was to analyze whether ouabain treatment also alters the structural and mechanical properties of mesenteric resistance arteries. Wistar rats were treated for 5 wk with ouabain (8.0 microg/day sc). The vascular structure and mechanics of the third-order branches of the mesenteric artery were assessed with pressure myography and confocal microscopy. Total collagen content was determined by picrosirius red staining, collagen I/III was analyzed by Western blot, and elastin was studied by confocal microscopy. Vascular reactivity was analyzed by wire myography. Internal and external diameters and cross sectional area were diminished, whereas the wall-to-lumen ratio was increased in arteries from ouabain-treated rats compared with controls. In addition, arteries from ouabain-treated rats were stiffer. Ouabain treatment decreased smooth muscle cell number and increased total and I/III collagens in the vascular wall. However, this treatment did not modify adventitia and media thickness, nuclei morphology, elastin structure, and vascular reactivity to norepinephrine and acetylcholine. The present work shows hypotrophic inward remodeling of mesenteric resistance arteries from ouabain-treated rats that seems to be the consequence of a combination of decreased cell number and impaired distension of the artery, possibly due to a higher stiffness associated with collagen deposition. The narrowing of resistance arteries could play a role in the pathogenesis of hypertension in this model. PMID- 16473963 TI - Left ventricular hypertrophy in mice with a cardiac-specific overexpression of interleukin-1. AB - Recent studies have identified the importance of proinflammatory cytokines in the development of left ventricular (LV) hypertrophy. However, the precise role of interleukin-1 (IL-1), one of the major proinflammatory cytokines, in the myocardium is not fully understood. In this study, we investigated the pathophysiological consequences of cardiac expression of IL-1 in vivo. We generated mice with a cardiac-specific overexpression of human IL-1alpha. We then analyzed their heart morphology and functions. Histological and echocardiographic analyses revealed concentric LV hypertrophy with preserved LV systolic function in the mice. Our results suggest that myocardial expression of IL-1 is sufficient to cause LV hypertrophy. PMID- 16473964 TI - Epoxyeicosatrienoic acids in cardioprotection: ischemic versus reperfusion injury. AB - Cytochrome P-450 (CYP) epoxygenases and their arachidonic acid (AA) metabolites, the epoxyeicosatrienoic acids (EETs), have been shown to produce increases in postischemic function via ATP-sensitive potassium channels (K(ATP)); however, the direct effects of EETs on infarct size (IS) have not been investigated. We demonstrate that two major regioisomers of CYP epoxygenases, 11,12-EET and 14,15 EET, significantly reduced IS in dogs compared to control (22.1 +/- 1.8%), whether administered 15 min before 60 min of coronary occlusion (6.4 +/- 1.9%, 11,12-EET; and 8.4 +/- 2.4%, 14.15-EET) or 5 min before 3 h of reperfusion (8.8 +/- 2.1%, 11,12-EET; and 9.7 +/- 1.4%, 14,15-EET). Pretreatment with the epoxide hydrolase metabolite of 14,15-EET, 14,15-dihydroxyeicosatrienoic acid, had no effect. The protective effect of 11,12-EET was abolished (24.3 +/- 4.6%) by the K(ATP) channel antagonist glibenclamide. Furthermore, one 5-min period of ischemic preconditioning (IPC) reduced IS to a similar extent (8.7 +/- 2.8%) to that observed with the EETs. The selective CYP epoxygenase inhibitor, N methylsulfonyl-6-(2-propargyloxyphenyl)hexanamide (MS-PPOH), did not block the effect of IPC. However, administration of MS-PPOH concomitantly with N methylsulfonyl-12,12-dibromododec-11-enanide (DDMS), a selective inhibitor of endogenous CYP omega-hydroxylases, abolished the reduction in myocardial IS expressed as a percentage of area at risk (IS/AAR) produced by DDMS (4.6 +/- 1.2%, DDMS; and 22.2 +/- 3.4%, MS-PPOH + DDMS). These data suggest that 11,12-EET and 14,15-EET produce reductions in IS/AAR primarily at reperfusion. Conversely, inhibition of CYP epoxygenases and endogenous EET formation by MS-PPOH, in the presence of the CYP omega-hydroxylase inhibitor DDMS blocked cardioprotection, which suggests that endogenous EETs are important for the beneficial effects observed when CYP omega-hydroxylases are inhibited. Finally, the protective effects of EETs are mediated by cardiac K(ATP) channels. PMID- 16473965 TI - SUGAR-DEPENDENT1 encodes a patatin domain triacylglycerol lipase that initiates storage oil breakdown in germinating Arabidopsis seeds. AB - Triacylglycerol hydrolysis (lipolysis) plays a pivotal role in the life cycle of many plants by providing the carbon skeletons and energy that drive postgerminative growth. Despite the physiological importance of this process, the molecular mechanism is unknown. Here, a genetic screen has been used to identify Arabidopsis thaliana mutants that exhibit a postgerminative growth arrest phenotype, which can be rescued by providing sugar. Seventeen sugar-dependent (sdp) mutants were isolated, and six represent new loci. Triacylglycerol hydrolase assays showed that sdp1, sdp2, and sdp3 seedlings are deficient specifically in the lipase activity that is associated with purified oil bodies. Map-based cloning of SDP1 revealed that it encodes a protein with a patatin-like acyl-hydrolase domain. SDP1 shares this domain with yeast triacylglycerol lipase 3 and human adipose triglyceride lipase. In vitro assays confirmed that recombinant SDP1 hydrolyzes triacylglycerols and diacylglycerols but not monoacylglycerols, phospholipids, galactolipids, or cholesterol esters. SDP1 is expressed predominantly in developing seeds, and a SDP1-green fluorescent protein fusion was shown to associate with the oil body surface in vivo. These data shed light on the mechanism of lipolysis in plants and establish that a central component is evolutionarily conserved among eukaryotes. PMID- 16473966 TI - The Arabidopsis SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE1 protein complex includes BRASSINOSTEROID-INSENSITIVE1. AB - Arabidopsis thaliana SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE1 (SERK1) is a leucine-rich repeat receptor-like kinase (LRR-RLK) involved in the acquisition of embryogenic competence and in male sporogenesis. To determine the composition of the SERK1 signaling complex in vivo, we generated plants expressing the SERK1 protein fused to cyan fluorescent protein under SERK1 promoter control. The membrane receptor complex was immunoprecipitated from seedlings, and the coimmunoprecipitating proteins were identified using liquid chromatography/matrix assisted laser desorption ionization-time of flight/mass spectrometry of the trypsin-released peptides. This approach identified two other LRR-RLKs, the BRASSINOSTEROID-INSENSITIVE1 (BRI1) receptor and its coreceptor, the SERK3 or BRI1-ASSOCIATED KINASE1 protein. In addition, KINASE-ASSOCIATED PROTEIN PHOSPHATASE, CDC48A, and 14-3-3nu were found. Finally, the MADS box transcription factor AGAMOUS-LIKE15 and an uncharacterized zinc finger protein, a member of the CONSTANS family, were identified as part of the SERK1 complex. Using blue native gel electrophoresis, we show that SERK1 and SERK3 are part of BRI1-containing multiple protein complexes with relative masses between 300 and 500 kD. The SERK1 mutant allele serk1-1 enhances the phenotype of the weak BRI1 allele bri1-119. Collectively, these results suggest that apart from SERK3, SERK1 is also involved in the brassinolide signaling pathway. PMID- 16473967 TI - The Arabidopsis-mei2-like genes play a role in meiosis and vegetative growth in Arabidopsis. AB - The Arabidopsis-mei2-Like (AML) genes comprise a five-member gene family related to the mei2 gene, which is a master regulator of meiosis in Schizosaccharomyces pombe and encodes an RNA binding protein. We have analyzed the AML genes to assess their role in plant meiosis and development. All five AML genes were expressed in both vegetative and reproductive tissues. Analysis of AML1-AML5 expression at the cellular level indicated a closely similar expression pattern. In the inflorescence, expression was concentrated in the shoot apical meristem, young buds, and reproductive organ primordia. Within the reproductive organs, strong expression was observed in meiocytes and developing gametes. Functional analysis using RNA interference (RNAi) and combinations of insertion alleles revealed a role for the AML genes in meiosis, with RNAi lines and specific multiple mutant combinations displaying sterility and a range of defects in meiotic chromosome behavior. Defects in seedling growth were also observed at low penetrance. These results indicate that the AML genes play a role in meiosis as well as in vegetative growth and reveal conservation in the genetic mechanisms controlling meiosis in yeast and plants. PMID- 16473968 TI - Arabidopsis REGULATOR OF AXILLARY MERISTEMS1 controls a leaf axil stem cell niche and modulates vegetative development. AB - Shoot branching is a major determinant of variation in plant stature. Branches, which form secondary growth axes, originate from stem cells activated in leaf axils. The initial steps by which axillary meristems (AMs) are specified and their stem cells organized are still poorly understood. We identified gain- and loss-of-function alleles at the Arabidopsis thaliana REGULATOR OF AXILLARY MERISTEMS1 (RAX1) locus. RAX1 is encoded by the Myb-like transcription factor MYB37 and is an Arabidopsis homolog of the tomato (Solanum lycopersicum) Blind gene. RAX1 is transiently expressed in a small central domain within the boundary zone separating shoot apical meristem and leaf primordia early in leaf primordium development. RAX1 genetically interacts with CUP-SHAPED COTYLEDON (CUC) genes and is required for the expression of CUC2 in the RAX1 expression domain, suggesting that RAX1 acts through CUC2. We propose that RAX1 functions to positionally specify a stem cell niche for AM formation. RAX1 also affects the timing of developmental phase transitions by negatively regulating gibberellic acid levels in the shoot apex. RAX1 thus defines a novel activity that links the specification of AM formation with the modulation of the rate of progression through developmental phases. PMID- 16473972 TI - Sleep in dementia. PMID- 16473973 TI - Diagnosis and treatment of sleep disorders in older adults. AB - Among the most common complaints of older adults are difficulty initiating or maintaining sleep. These problems result in insufficient sleep at night, which then results in an increased risk of falls, difficulty with concentration and memory, and overall decreased quality of life. Difficulties sleeping, however, are not an inevitable part of aging. Rather, these sleep complaints are often secondary to medical and psychiatric illness, the medications used to treat these illnesses, circadian rhythm changes, or other sleep disorders. The task for the geriatric psychiatrist is to identify the causes of these complaints and then initiate appropriate treatment. PMID- 16473969 TI - Arabidopsis PEN3/PDR8, an ATP binding cassette transporter, contributes to nonhost resistance to inappropriate pathogens that enter by direct penetration. AB - Arabidopsis thaliana is a host to the powdery mildew Erysiphe cichoracearum and nonhost to Blumeria graminis f. sp hordei, the powdery mildew pathogenic on barley (Hordeum vulgare). Screening for Arabidopsis mutants deficient in resistance to barley powdery mildew identified PENETRATION3 (PEN3). pen3 plants permitted both increased invasion into epidermal cells and initiation of hyphae by B. g. hordei, suggesting that PEN3 contributes to defenses at the cell wall and intracellularly. pen3 mutants were compromised in resistance to the necrotroph Plectosphaerella cucumerina and to two additional inappropriate biotrophs, pea powdery mildew (Erysiphe pisi) and potato late blight (Phytophthora infestans). Unexpectedly, pen3 mutants were resistant to E. cichoracearum. This resistance was salicylic acid-dependent and correlated with chlorotic patches. Consistent with this observation, salicylic acid pathway genes were hyperinduced in pen3 relative to the wild type. The phenotypes conferred by pen3 result from the loss of function of PLEIOTROPIC DRUG RESISTANCE8 (PDR8), a highly expressed putative ATP binding cassette transporter. PEN3/PDR8 tagged with green fluorescent protein localized to the plasma membrane in uninfected cells. In infected leaves, the protein concentrated at infection sites. PEN3/PDR8 may be involved in exporting toxic materials to attempted invasion sites, and intracellular accumulation of these toxins in pen3 may secondarily activate the salicylic acid pathway. PMID- 16473970 TI - FLOWERING LOCUS C mediates natural variation in the high-temperature response of the Arabidopsis circadian clock. AB - Temperature compensation contributes to the accuracy of biological timing by preventing circadian rhythms from running more quickly at high than at low temperatures. We previously identified quantitative trait loci (QTL) with temperature-specific effects on the circadian rhythm of leaf movement, including a QTL linked to the transcription factor FLOWERING LOCUS C (FLC). We have now analyzed FLC alleles in near-isogenic lines and induced mutants to eliminate other candidate genes. We showed that FLC lengthened the circadian period specifically at 27 degrees C, contributing to temperature compensation of the circadian clock. Known upstream regulators of FLC expression in flowering time pathways similarly controlled its circadian effect. We sought to identify downstream targets of FLC regulation in the molecular mechanism of the circadian clock using genome-wide analysis to identify FLC-responsive genes and 3503 transcripts controlled by the circadian clock. A Bayesian clustering method based on Fourier coefficients allowed us to discriminate putative regulatory genes. Among rhythmic FLC-responsive genes, transcripts of the transcription factor LUX ARRHYTHMO (LUX) correlated in peak abundance with the circadian period in flc mutants. Mathematical modeling indicated that the modest change in peak LUX RNA abundance was sufficient to cause the period change due to FLC, providing a molecular target for the crosstalk between flowering time pathways and circadian regulation. PMID- 16473974 TI - Comparison between informant-observed and actigraphic assessments of sleep-wake rhythm disturbances in demented residents of homes for the elderly. AB - OBJECTIVE: Sleep-wake rhythm disturbances frequently occur in demented elderly and are of clinical relevance because they herald accelerated functional decline and institutionalization. Assessment of sleep-wake rhythm disorders is therefore of significant importance and can be performed by questionnaires or actigraphy, i.e., the recording of wrist activity. The present study investigates the relation of these two types of measurement by simultaneously assessing actigraphy and the Circadian Sleep Inventory for Normal and Pathological States (CSINAPS). METHODS: Seventy-eight elderly subjects, mean age 85+/-6 years, living in group care facilities of 12 homes for the elderly, wore an actigraph for two weeks. Caregivers completed the nurse informant CSINAPS. Spearman rank correlations and Mann-Whitney U tests were calculated over the equivalent sleep-wake rhythm parameters as derived from actigraphy and from the CSINAPS. RESULTS: Good correlations were found between questionnaire items about habitual timing of sleep and wakefulness and their actigraphic counterparts. Caregivers overestimated the actual sleep time between sleep onset and offset by 96 minutes. Questionnaire reports of sleep disturbances like wandering at night were also reflected in actigraphy parameters. However, the questionnaire and actigraphy variables correlate only modestly and may complement each other. In our study, both actigraphy and the CSINAPS seemed to miss the previously established high prevalence of sleep-disordered breathing (SDB) and leg movements during sleep (LM). CONCLUSION: The assessment of sleep and wake disturbances in demented elderly is best served by parallel use of a questionnaire like the CSINAPS and actigraphy. Moreover, if SDB and LM are a focus of interest, additional assessments are needed. PMID- 16473976 TI - Daytime sleeping, sleep disturbance, and circadian rhythms in the nursing home. AB - OBJECTIVE: This study reports the frequency of abnormal daytime sleeping and identifies factors related to daytime sleeping, nighttime sleep disturbance, and circadian rhythm abnormalities among nursing home residents. METHODS: The authors conducted secondary analysis of data collected under usual care conditions within a nonpharmacologic sleep intervention trial. All residents from four Los Angeles nursing homes were screened for daytime sleeping (asleep>or=15% of observations, 9:00 am-5:00 pm). Consenting residents with daytime sleeping had two nights of wrist actigraphy to assess nighttime sleep disturbance (asleep<80%, 10:00 pm-6:00 am). Residents with nighttime sleep disturbance completed an additional 72-hour wrist actigraphy recording to assess circadian activity rhythms and light exposure. RESULTS: Sixty-nine percent of 492 observed residents had daytime sleeping, of whom 60% also had disturbed nighttime sleep. Sleep disturbance and daytime sleeping were rarely documented in medical records. Residents spent one third of the day in their rooms, typically in bed, and were seldom outdoors or exposed to bright light. More time in bed and less social activity were significant predictors of daytime sleepiness. Ninety-seven percent of residents assessed had abnormal circadian rhythms. More daytime sleeping and less nighttime sleep were associated with weaker circadian activity rhythms. Later circadian rhythm acrophase (peak) was associated with more bright light exposure. CONCLUSION: Daytime sleepiness, nighttime sleep disturbance, and abnormal circadian rhythms were common in nursing home residents. Modifiable factors (e.g., time in bed) are associated with sleep/wake abnormalities. Mental health specialists should consider the complexity of factors causing sleep problems in nursing home residents. PMID- 16473975 TI - Factors associated with caregiver reports of sleep disturbances in persons with dementia. AB - OBJECTIVE: This investigation examined the relationship between caregivers' reports of sleep disturbances in persons with Alzheimer disease (AD) and actigraphic records of patients' sleep-wake activity, and explored the factors associated with discrepancies in this relationship. METHOD: Forty-six patients with AD living with their caregivers participated. Before study entry, all caregivers reported poor patient sleep quality using the Neuropsychiatric Inventory (NPI) Nighttime Behavior Scale. Patient sleep-wake activity was recorded for one week using an Actillume wrist-movement recorder. RESULTS: Although all patients were reported by caregivers to have multiple sleep disturbances, 41% of patients had actigraphic sleep efficiencies in the normal range and 43% averaged eight or more hours of sleep nightly. In bivariate analyses, greater patient percent nighttime sleep (indicative of more discrepancy between subjective reports and objective outcomes) was associated with less patient cognitive, physical and functional impairment, lower self-rated depression, higher self-rated quality of life, and less daytime sleepiness. No patient variable significantly predicted total patient nocturnal sleep time. In both bivariate and multivariate analyses, greater patient percent sleep and total sleep time were also associated with caregiver factors, particularly greater use of criticism as a behavior management strategy. CONCLUSION: The study suggests that both patient and caregiver factors contribute to reported sleep problems in community-dwelling patients with AD, and that caregiver reports and objective sleep assessments frequently may not agree. Treatments should consider these dyadic contributions rather than focusing on caregiver reports of patient symptoms alone. PMID- 16473978 TI - Mapping scores onto stages: mini-mental state examination and clinical dementia rating. AB - OBJECTIVE: Although the clinical course of Alzheimer disease (AD) is gradual, it is useful for a number of reasons to distinguish between different levels of severity. The Clinical Dementia Rating (CDR) has demonstrated high validity and reliability for this purpose, but it requires a considerable amount of data to be collected both from the patient and from an informant. In the present study, the authors mapped Mini-Mental State Examination (MMSE) scores onto CDR categories to determine how well the MMSE performs as a surrogate of the CDR as a timesaving method of staging dementia. METHOD: Eight hundred sixty-three probands, including 524 patients with probable AD, 92 patients with questionable dementia, and 247 with memory complaints but no objective cognitive impairment, were included. Cutoff scores were identified on one-half of the sample using a receiver operating characteristic analysis. The cutoff values were then applied to the other half of the sample, and the agreement between MMSE score ranges and CDR stages was determined by calculating Cohen's kappa. RESULTS: The MMSE discriminated well between CDR stages 0.5, 1, 2, and 3 but performed poorly in the separation between CDR stages zero and 0.5. The MMSE ranges were 30 for no, 26-29 for questionable, 21-25 for mild, 11-20 for moderate, and 0-10 for severe dementia. Substantial agreement between the two instruments was obtained for the categories mild (kappa=0.62, p<0.001, N=115), moderate (kappa=0.69, p<0.001, N=114), and severe dementia (kappa=0.76, p<0.001, N=39), whereas the agreement was moderate for no (kappa=0.44, p<0.001, N=120) and only fair for questionable dementia (kappa=0.28, p<0.001, N=42). CONCLUSION: The MMSE can be used as a surrogate measure for the CDR for the staging of dementia in AD. PMID- 16473977 TI - Combination of clinical and neuropsychologic information as a better predictor of the progression to Alzheimer disease in questionable dementia individuals. AB - OBJECTIVES: This study aimed to examine the clinical outcomes of questionable dementia (QD) elderly subjects after three years of follow-up and to compare the ability of a standardized clinical assessment, neuropsychologic tests, the ApoE genotyping, and possible combinations of these methods to predict their progression to Alzheimer disease (AD). METHODS: One hundred six elderly subjects with QD were evaluated with a standardized clinical assessment, neuropsychologic tests, and ApoE genotyping and followed up annually. The Clinical Dementia Rating Sum of Boxes (CDR-SOB) score was used as a quantitative summary score of the standardized clinical assessment on the overall functioning of the subjects. RESULTS: Among the individuals remaining in the study after the 3-year follow-up period, 8.3% had improved to a state of normal cognition, 72.7% were still in the QD state. and 19.4% had progressed to clinically evident AD. Although each of CDR SOB, Word List Recall (WLR), and ApoE epsilon4 genotype was predictive for AD, the combination of CDR-SOB and WLR was found to predict AD better than any single variable. However, the addition of the ApoE epsilon4 genotype information to CDR SOB or WLR did not improve their predictive ability. CONCLUSION: The combination of clinical assessment on function and episodic memory test can improve the predictive ability of each measure for progression to AD in QD individuals. However, ApoE genotyping dose not make an additional contribution to AD prediction in QD individuals when used in combination with clinical assessment or memory test. PMID- 16473979 TI - Cerebrovascular risk factors, executive dysfunction, and depression in older primary care patients. AB - OBJECTIVE: "Executive" cognitive functions may be of particular clinical importance in geriatric depression and may reflect underlying cerebrovascular disease. This study examined the associations of selected components of executive function with cerebrovascular risk factors, depression, and overall functional status. METHOD: Study measures were completed on 448 primary care patients aged>or=65 years based on patient interviews and medical chart review. Multiple regression techniques determined the presence of specified independent associations. RESULTS: Some but not all study hypotheses were confirmed. Cerebrovascular risk factors were associated with major depression and with some cognitive measures, but their associations with depression and with the most specific measures of executive function were limited and not independent of overall medical burden. Measures of initiation-perseveration and mental set shifting were associated with overall functional disability; these cognitive measures were not associated with depression diagnosis, or with depressive symptoms when also covarying medical burden or excluding patients with dementia. CONCLUSIONS: Clinicians should be aware of the potential functional significance of these components of cognition. Longitudinal risk factor studies and complementary techniques such as neuroimaging may help identify pathogenetically distinct subgroups of later-life depression that might respond preferentially to specific interventions. PMID- 16473980 TI - Characteristics of visual hallucinations in Parkinson disease dementia and dementia with lewy bodies. AB - OBJECTIVE: Parkinson disease dementia (PDD) and dementia with Lewy bodies (DLB) have overlapping clinical and pathologic features. Recurrent visual hallucinations (RVH) are common in both disorders. The authors have compared details of hallucination characteristics and associated neuropsychiatric features in DLB and PDD. METHODS: This is a descriptive, cross-sectional study using the Institute of Psychiatry Visual Hallucinations Interview (IP-VHI) to explore self reported frequency, duration, and phenomenology of RVH in PDD and DLB. The caregivers' ratings of hallucinations and other neuropsychiatric features were elicited with the Neuropsychiatric Inventory (NPI). RESULTS: Fifty-six patients (35 PDD; 21 DLB) with RVH were assessed. Hallucination characteristics were similar in both disorders. Simple hallucinations were rare. Most patients experienced complex hallucinations daily, normally lasting minutes. They commonly saw people or animals and the experiences were usually perceived as unpleasant. NPI anxiety scores were higher in PDD. Neuropsychiatric symptoms coexisting with hallucinations were apathy, sleep disturbance, and anxiety. CONCLUSIONS: Patients with mild to moderate dementia can provide detailed information about their hallucinations. Characteristics of RVH were similar in PDD and DLB, and phenomenology suggests the involvement of dorsal and ventral visual pathways in their generation. The coexistence of RVH with anxiety, apathy, and sleep disturbance is likely to impair patients' quality of life and may have treatment implications. PMID- 16473981 TI - Daily affect in Parkinson disease is responsive to life events and motor symptoms. AB - OBJECTIVE: The aims of this study were to examine the daily affective experiences of patients with Parkinson disease (PD) and to determine their association with daily events and motor symptoms. Specifically, it was intended to test the hypothesis that PD, even in the absence of depression, is associated with anhedonia. METHOD: Nondepressed male subjects with PD (N=24) and a comparison group of healthy elderly males (N=23) completed daily affect rating scales and, for the patients with PD, a supplemental self-assessment questionnaire of PD related symptoms for 4 consecutive weeks. The effect of daily events and PD related symptoms on daily affect was examined using linear and logistic mixed regression models. RESULTS: Overall, patients with PD reported significantly less positive and more negative affect than healthy peers over time. There were similar, and expected, associations between negative events and affect in both groups. Although patients with PD reported far fewer positive events than control subjects, they reported as great an improvement in affect in response to them. Regarding self-reported PD-related symptoms, only increasing severity of core motor symptoms was independently associated with worse affect. CONCLUSIONS: Although the conclusions of this study are tempered by a comparison group that is not optimal, our results suggest that patients with PD do not demonstrate anhedonia in response to positive life events. The gross intergroup difference in daily events suggests the potential value of interventions that emphasize daily engagement in positive experiences to improve positive affective tone. PMID- 16473982 TI - Test characteristics of the 15-item geriatric depression scale and Hamilton depression rating scale in Parkinson disease. AB - OBJECTIVE: The objective of this study was to compare the sensitivity, specificity, and diagnostic accuracy of the 15-item Geriatric Depression Scale (GDS-15) and the Hamilton Depression Rating Scale (HDRS) in patients with Parkinson disease (PD). METHOD: A convenience sample of 148 outpatients with idiopathic PD receiving specialty care completed the GDS-15 and were administered the HDRS and Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (SCID) depression module by a research psychiatrist or trained research assistant. Receiver-operating characteristic (ROC) curves were plotted for the GDS-15 and HDRS scores with a SCID diagnosis of a depressive disorder as the state variable. RESULTS: Thirty-two subjects (22%) were diagnosed with a depressive disorder. The discriminant validity of the GDS 15 and HDRS were both high (ROC area under the curve: 0.92 and 0.91, respectively), with greatest dichotomization for the GDS-15 at a cutoff of 4/5 (87% accuracy, 88% sensitivity, 85% specificity) and the HDRS at a cutoff of 9/10 (83% accuracy, 88% sensitivity, 78% specificity). CONCLUSIONS: The GDS-15 performs well as a screening instrument and in distinguishing depressed from nondepressed patients in PD. Its test characteristics are comparable to the HDRS. Because it is a brief instrument and can be self-administered, it is an excellent depression screening tool in this population. PMID- 16473983 TI - Adherence to continuous positive airway pressure treatment in patients with Alzheimer's disease and obstructive sleep apnea. AB - OBJECTIVE: This analysis examined whether patients with Alzheimer disease (AD) tolerate continuous positive airway pressure (CPAP). METHOD: Thirty patients with AD were randomized to CPAP or sham CPAP and completed sleep, depression, and quality-of-life questionnaires. Participants could choose to continue treatment after the trial. RESULTS: Patients wore CPAP for 4.8 hours per night. More depressive symptoms were associated with worse adherence (rS=-0.37; N=30, p<0.04). Patients who continued using CPAP had fewer depressive symptoms (t [19]=2.45, p=0.02) and better adherence (t [19]=2.32, p=0.03) during the trial. CONCLUSION: Patients with AD with obstructive sleep apnea can tolerate CPAP. Adherence and long-term use may be more difficult among those patients with more depressive symptoms. PMID- 16473985 TI - Emotional intelligence and psychiatric training. PMID- 16473984 TI - Methylphenidate-enhanced antidepressant response to citalopram in the elderly: a double-blind, placebo-controlled pilot trial. AB - OBJECTIVE: The authors evaluated the potential of methylphenidate to accelerate and enhance antidepressant response to citalopram in elderly depressed patients. METHODS: Sixteen outpatients with major depression were treated in a 10-week double-blind trial. Response was defined as a score on the Hamilton Depression Rating Scale (24-item) of less than 10. RESULTS: An accelerated response was observed by week 3 in five subjects receiving citalopram (CIT)+methylphenidate (MPH) and in none of those receiving CIT+placebo (PBO). Subjects receiving citalopram and methylphenidate showed a significant improvement in depressive symptoms compared with those on citalopram and placebo. CONCLUSION: Combined treatment with citalopram and methylphenidate appears to be a viable strategy for accelerating and enhancing antidepressant response in elderly depressed patients limited by tolerability and safety. PMID- 16473987 TI - Financial and organizational turmoil in the academic health center: is it a crisis or an opportunity for medical education? PMID- 16473988 TI - Mental health disparities, diversity, and cultural competence in medical student education: how psychiatry can play a role. AB - OBJECTIVE: The authors review recent developments in healthcare policy, including eliminating disparities in mental healthcare, increasing diversity in the healthcare workforce, and cultural competence. Following a discussion of the Liaison Committee on Medical Education (LCME) standards, as they relate to disparity, cultural competence, and diversity, the authors discuss an action plan describing the role of psychiatry in addressing these issues. METHODS: Key policy documents are reviewed for disparities, cultural competence, and diversity in healthcare and then in mental health specifically. RESULTS: Important developments in healthcare policy regarding these areas have occurred. CONCLUSION: Psychiatry can play a vital role in addressing disparities, cultural competence, and diversity in medical student education. PMID- 16473989 TI - Medical school research pipeline: medical student research experience in psychiatry. AB - OBJECTIVE: The authors discuss the importance of introducing research training in psychiatry and neurosciences to medical students. METHODS: A review of existing models of research training in psychiatry with focus on those providing research training to medical students is presented. RESULTS: Two research-training models for medical students that are easy to adopt and have possible nationwide applicability are identified, along with other useful research-training models. CONCLUSION: Specific recommendations on how to foster research training in psychiatry and neurosciences for medical students throughout the U.S. are presented. PMID- 16473990 TI - From Yoda to Sackett: the future of psychiatry medical student education. AB - OBJECTIVE: The authors discuss approaches to curricular goals, methods, and assessments in the education of medical students in psychiatry. METHODS: Using current educational principles and opinions on curricular reform in medical student education, an outline for a core curriculum and an individualized approach to medical student education were developed. RESULTS: A curricular outline addressing both content and organization was developed for all students as well as those specifically entering clinical psychiatry, neuroscience research, and primary care. CONCLUSIONS: In addition to a standard core curriculum there should be a tailored approach for certain subsets of students. The core curriculum should be an "ideal minimum" that recognizes the importance of evidence based medicine and is strategically planned and learner focused. PMID- 16473991 TI - APA Summit on Medical Student Education Task Force on Informatics and Technology: learning about computers and applying computer technology to education and practice. AB - OBJECTIVE: This article provides a brief overview of important issues for educators regarding medical education and technology. METHODS: The literature describes key concepts, prototypical technology tools, and model programs. A work group of psychiatric educators was convened three times by phone conference to discuss the literature. Findings were presented to and input was received from the 2005 Summit on Medical Student Education by APA and the American Directors of Medical Student Education in Psychiatry. RESULTS: Knowledge of, skills in, and attitudes toward medical informatics are important to life-long learning and modern medical practice. A needs assessment is a starting place, since student, faculty, institution, and societal factors bear consideration. Technology needs to "fit" into a curriculum in order to facilitate learning and teaching. CONCLUSION: Learning about computers and applying computer technology to education and clinical care are key steps in computer literacy for physicians. PMID- 16473992 TI - Medical education and the pharmaceutical industry: managing an uneasy alliance. PMID- 16473993 TI - The role of the pharmaceutical industry in medical education in psychiatry. PMID- 16473994 TI - The pharmaceutical role. PMID- 16473995 TI - The ethical commitments of academic faculty in psychiatric education. AB - OBJECTIVE: This article explores the commitment of faculty to ethics training in psychiatric education. Although psychiatry has insufficiently addressed the profession's need for ethics training in education, program directors acknowledge its critical importance, and its positive impact has been demonstrated. Additionally, residents often seek ethics training as part of their instruction. METHOD: The author suggests that academic faculty could respond to the profession's inadequate treatment of ethics training by helping trainees develop moral agency--the ability to recognize, assess, and respond to ethical dilemmas; decide what constitutes right or wrong care; and act accordingly. The author also describes how this objective could be met by promoting professionalism and offering didactic instruction that address substantive and process issues regarding psychiatric care. CONCLUSION: Specific recommendations are provided. PMID- 16473996 TI - A developmental model for enhancing research training during psychiatry residency. AB - OBJECTIVE: The authors describe a developmental model for enhancing residency research training for careers in academic psychiatry. Over the past 10 years, the University of Pittsburgh Department of Psychiatry has developed a research track (RT) for its residents. While the Department's plan has been to address the critical need of training physician-scientists in psychiatry, the RT continues to evolve as a structured extension of the University's residency-training program. Recently, the University's departmental leadership has taken several steps that address regulatory, institutional, and personal barriers to residency research training put forth by the 2003 Institute of Medicine (IOM) report. METHODS: The authors outline a model of residency research training, elements of which should be exportable to the majority of U.S. psychiatry residency programs. RESULTS: For residents in the RT, up to 50% of time in PGY-3 and up to 75% of time in PGY-4 can be devoted to research-related activities. The authors currently have 13 residents and fellows in their track. Over the past 10 years, 15 of 33 RT residents have become research postdocs or full-time grant-funded researchers in academic positions. CONCLUSION: The authors' experience suggests that it is possible to organize and implement an RT during psychiatry residency within the parameters presented by the Psychiatry Residency Review Committee (RRC). PMID- 16473997 TI - Practice of dialectical behavior therapy after psychiatry residency. AB - OBJECTIVE: The University of Washington (UW) psychiatry residency program attempted to determine how participation in a dialectical behavior therapy (DBT) training program influenced the practice of its graduates. METHODS: A survey was completed by 30 graduates who participated in elective DBT training. This survey obtained information about their DBT training and their current use of DBT. RESULTS: Twenty seven percent of the respondents have performed "formal" DBT since graduation. The rest reported using at least some DBT interventions. The "dose" of DBT training received had an influence on the number of DBT interventions used. CONCLUSIONS: These findings suggest that DBT training influenced psychiatrists' use of DBT in their practice. PMID- 16474000 TI - Acoustics. PMID- 16473998 TI - Family skills for general psychiatry residents: meeting ACGME core competency requirements. AB - OBJECTIVE: The authors discuss the knowledge, attitudes, and skills needed for a resident to be competent in supporting and working with families, as mandated by the residency review committee (RRC) core competencies. METHODS: The RRC core competencies, as they relate to patients and their families, are reviewed. The Group for Advancement of Psychiatry (GAP) presents an operational version of these core competencies. RESULTS: Methods of assessment, challenges in teaching, and ways of overcoming programmatic constraints are outlined. Examples of training programs that offer ways of integrating the teaching of family skills into existing programs are described. CONCLUSIONS: The implications of the current RRC core competency requirements pertaining to families have the potential to change the training environment substantially. The GAP Family Committee proposes recommendations to facilitate the training of residents in family skills. PMID- 16474001 TI - A research agenda concerning depictions of mental illness in children's media. AB - OBJECTIVE: To review research on depictions of mental illness in mass media directed to children and to identify requirements for further research in this important field. METHODS: The authors identified published research on depictions of mental illness in children's media and the important strengths and weaknesses of such research. RESULTS: Only four relevant studies and one collection of reviews of children's books were found. Three of the studies focused on depictions of mental illness in film, and one study focused on depictions of mental illness in television. While some depictions evoked sympathy, others were frightening and denigrating. There was often insufficient data necessary to ascertain how researchers engaged with the language and visual imagery of the medium. CONCLUSIONS: There is an insufficient amount of research in this area. In particular, there is a need to determine how children attribute meaning to depictions of mental illness in children's media. PMID- 16474002 TI - Opposite effects of wortmannin and 2-(4-morpholinyl)-8-phenyl-1(4H)-benzopyran-4 one hydrochloride on toll-like receptor-mediated nitric oxide production: negative regulation of nuclear factor-{kappa}B by phosphoinositide 3-kinase. AB - A number of previous studies have suggested the involvement of phosphoinositide 3 kinase (PI3K) in Toll-like receptor (TLR) signaling. However, there have also been a number of conflicting reports. The PI3K inhibitor wortmannin greatly enhanced TLR-mediated inducible nitric-oxide synthase (iNOS) expression and cytokine production in the mouse macrophage cell line Raw264.7. The effect of wortmannin was common to TLR2, -3, -4, and -9 and was accompanied by activation of nuclear factor-kappaB and up-regulation of cytokine mRNA production. We were surprised to find that another PI3K inhibitor, LY294002, strongly suppressed the production of iNOS and cytokines. This effect of 2-(4-morpholinyl)-8-phenyl-1(4H) benzopyran-4-one hydrochloride (LY294002) was based on its inhibitory effect on mRNA synthesis. Expression of dominant-negative mutants of PI3K in macrophages augmented the lipopolysaccharideinduced expression of iNOS. Introduction of a pH1 vector producing short hairpin RNA that targets a catalytic subunit of PI3K (p110beta) also enhanced the TLR-mediated responses. Thus, the augmentation of TLR signals by wortmannin was mediated through the inhibition of PI3K, whereas the effect of LY294002 was not explained by its effect on PI3K. These discrepancies in the effects of pharmacological inhibitors in TLR-signaling may have caused confusion regarding the role of PI3K in innate immunity. PMID- 16474003 TI - Molecular determinants of HERG channel block. AB - Drug-induced block of cardiac hERG K+ channels causes acquired long QT syndrome. Here, we characterized the molecular mechanism of hERG block by two low-potency drugs (Nifekalant and bepridil) and two high-potency drugs 1-[2-(6-methyl 2pyridyl)ethyl]-4-(4-methylsulfonyl aminobenzoyl)piperidine (E-4031) and dofetilide). Channels were expressed in Xenopus laevis oocytes, and currents were measured using the two-microelectrode voltage-clamp technique. All four drugs progressively reduced hERG current during a 20-s depolarization to 0 mV after a 10-min pulse-free period, consistent with the preferential block of open channels. Recovery from block in response to pulses to -160 mV was observed for D540K hERG channels but not for wild-type hERG channels, suggesting that all four drugs are trapped in the central cavity by closure of the activation gate. The molecular determinants of hERG channel block were defined by using a site directed mutagenesis approach. Mutation to alanine of three residues near the pore helix (Thr623, Ser624, and Val625) and four residues in Ser6 (Gly648, Tyr652, Phe656, and Val659) reduced channel sensitivity to block by dofetilide and E-4031, effects identical with those reported previously for two other methanesulfonanilides, (+)- N -[1' -(6-cyano-1,2,3,4-tetrahydro-2(R) naphthalenyl)-3,4-dihydro-4(R)-hydroxyspiro(2H -1-benzopyran-2,4' -piperidin)-6 yl]-methanesulfonamide] monohydrochloride (MK-499) and ibutilide. The effect of nifekalant on mutant channels was similar, except that V659A retained normal sensitivity and I655A channels were less sensitive. Finally, mutation of the three residues near the pore helix and Phe656 in the Ser6 domain reduced channel block by bepridil. We conclude that the binding site is not identical for all drugs that preferentially block hERG in the open state. PMID- 16474004 TI - Effects of a novel cognitive enhancer, spiro[imidazo-[1,2-a]pyridine-3,2-indan] 2(3H)-one (ZSET1446), on learning impairments induced by amyloid-beta1-40 in the rat. AB - We have previously shown that intracerebroventricular (i.c.v.) infusion of amyloid-beta (Abeta)1-40 produces oxidative stress and cholinergic dysfunction, as well as learning and memory deficits, in rats. In the present study, effects of a newly synthesized azaindolizinone derivative, spiro[imidazo[1,2-a]pyridine 3,2-indan]-2(3H)-one (ZSET1446), were assessed in rats with learning deficits induced by Abeta1-40 or scopolamine. The i.c.v. infusion of Abeta1-40 caused impairments in spontaneous alternation behavior in a Y-maze task, spatial reference and short-term memory in a water-maze task, and retention of passive avoidance learning. Abeta1-40-infused rats also showed reduction in choline acetyltransferase (ChAT) activity in the medial septum and hippocampus, but not in the basal forebrain and cortex, and a decrease in glutathione S-transferase (GST)-like immunoreactivity in the cortex. Nicotine-stimulated acetylcholine (ACh) release in Abeta1-40-infused rats was lower than that in vehicle-infused rats. Oral administration of ZSET1446 at the dose range of 0.01 to 1 mg/kg ameliorated Abeta1-40-induced learning impairment in Y-maze, water-maze, and passive-avoidance tasks. ZSET1446 reversed the decrease of ChAT activity in the medial septum and hippocampus, GST-like immunoreactivity in the cortex, and nicotine-stimulated ACh release of Abeta1-40-treated rats to the levels of vehicle-infused control rats. Furthermore, 0.001 to 0.1 mg/kg ZSET1446 showed ameliorative effects on learning impairments caused by scopolamine in a passive avoidance task. These results suggest that ZSET1446 may be a potential candidate for development as a therapeutic agent to manage cognitive impairment associated with conditions such as Alzheimer's disease. PMID- 16474005 TI - Differential role of cyclooxygenase 1 and 2 isoforms in the modulation of colonic neuromuscular function in experimental inflammation. AB - This study examines the role played by cyclooxygenase (COX) isoforms (COX-1 and 2) in the regulation of colonic neuromuscular function in normal rats and after induction of colitis by 2,4-dinitrobenzenesulfonic acid (DNBS). The expression of COX-1 and COX-2 in the colonic neuromuscular layer was assessed by reverse transcription-polymerase chain reaction and immunohistochemistry. The effects of COX inhibitors on in vitro motility were evaluated by studying electrically induced and carbachol-induced contractions of the longitudinal muscle. Both COX isoforms were constitutively expressed in normal colon; COX-2 was up-regulated in the presence of colitis. In normal and inflamed colon, both COX isoforms were mainly localized in neurons of myenteric ganglia. In the normal colon, indomethacin (COX-1/COX-2 inhibitor), SC-560 [5-(4-chloro-phenyl)-1-(4 methoxyphenyl)-3-trifluoromethylpyrazole] (COX-1 inhibitor), or DFU [5,5-dimethyl 3-(3-fluorophenyl)-4-(4-methylsulfonyl)phenyl-2(5H)-furanone] (COX-2 inhibitor) enhanced atropine-sensitive electrically evoked contractions. The most prominent effects were observed with indomethacin or SC-560 plus DFU. In the inflamed colon, SC-560 lost its effect, whereas indomethacin and DFU maintained their enhancing actions. These results were more evident after blockade of noncholinergic pathways. In rats with colitis, in vivo treatment with superoxide dismutase or S-methylisothiourea (inhibitor of inducible nitric-oxide synthase) restored the enhancing motor effect of SC-560. COX inhibitors had no effect on carbachol-induced contractions in normal or DNBS-treated rats. In conclusion, in the normal colon, both COX isoforms act at the neuronal level to modulate the contractile activity driven by excitatory cholinergic pathways. In the presence of inflammation, COX-1 activity is hampered by oxidative stress, and COX-2 seems to play a predominant role in maintaining an inhibitory control of colonic neuromuscular function. PMID- 16474007 TI - Iron Imports. V. Transport of iron through the intestinal epithelium. AB - Iron absorption across the brush-border membrane requires divalent metal transporter 1 (DMT1), whereas ferroportin (FPN) and hephaestin are required for exit across the basolateral membrane. However, how iron passes across the enterocyte is poorly understood. Both chaperones and transcytosis have been postulated to account for intracellular iron transport. With iron feeding, DMT1 undergoes endocytosis and FPN translocates from the apical cytosol to the basolateral membrane. The fluorescent metallosensor calcein offered to the basolateral surface of enterocytes is found in endosomes in the apical compartment, and its fluorescence is quenched when iron is offered to the apical surface. These experiments are consistent with vesicular iron transport as a possible pathway for intracellular iron transport. PMID- 16474008 TI - Effect of aging on bolus kinematics during the pharyngeal phase of swallowing. AB - Swallowing difficulty is a common complaint in the elderly and, although there are data for the biomechanics of liquid swallows, little is known about solid bolus motion, or kinematics, in the elderly. The aims of this study were as follows: 1) to characterize and compare solid and liquid bolus kinematics in the elderly and compare the findings with those in young subjects and 2) to correlate bolus kinematics and dynamics. Concurrent manometric-fluoroscopic techniques were used to study eight young and eight elderly subjects. The subjects performed four swallows each of 0.2-cm-diameter solid barium pellets and 5 ml of liquid barium during sagittal fluoroscopy and six-channel pharyngoesophageal manometry. Images were digitized for analysis of kinematic properties such as velocity and acceleration. Dynamic pressures were recorded and coordinated with kinematic events. Image analysis showed that velocity varied as the pellet passed through the hypopharynx, pharynx, and upper esophageal sphincter. In young subjects, pellet kinematics were characterized by two zones of pellet acceleration: one over the tongue base and another as the pellet passed through the upper esophageal sphincter. Although the elderly showed a similar zone of acceleration over the base of the tongue, the second zone of pellet acceleration was not seen. Decreasing pressure gradients immediately distal to the position of the solid pellet and liquid bolus characterized dynamics for all subjects. This decreasing pressure gradient was significantly larger in elderly than in young subjects. Bolus kinematics and dynamics were significantly altered among elderly compared with young subjects. Among these differences were the absence of hypopharyngeal bolus acceleration and a significant increase in the trans-sphincteric pressure gradient in the elderly. PMID- 16474009 TI - Molecular mechanism of tumor necrosis factor-alpha modulation of intestinal epithelial tight junction barrier. AB - A TNF-alpha-induced increase in intestinal epithelial tight junction (TJ) permeability has been proposed to be an important proinflammatory mechanism contributing to intestinal inflammation in Crohn's disease and other inflammatory conditions. Previous studies from our laboratory suggested that the TNF-alpha induced increase in intestinal TJ permeability was mediated by an increase in myosin light chain kinase (MLCK) protein expression. However, the molecular mechanisms that mediate the TNF-alpha increase in intestinal TJ permeability and MLCK protein expression remain unknown. The purpose of this study was to delineate the intracellular and molecular mechanisms that mediate the TNF-alpha induced increase in intestinal TJ permeability; using an in vitro intestinal epithelial model system consisting of filter-grown Caco-2 intestinal epithelial monolayers. To examine the molecular mechanisms involved in the TNF-alpha regulation of intestinal TJ barrier, we identified and cloned for the first time a functionally active MLCK promoter region. TNF-alpha treatment of filter-grown Caco-2 monolayers transfected with plasmid vector containing the MLCK promoter region produced an increase in MLCK promoter activity and MLCK transcription. The TNF-alpha-induced increase in MLCK transcription corresponded to a sequential increase in MLCK protein expression, MLCK activity, and Caco-2 TJ permeability. The TNF-alpha-induced increase in MLCK promoter activity was mediated by NF kappaB activation, and the inhibition of NF-kappaB activation prevented the TNF alpha-induced increase in promoter activity and the subsequent increase in MLCK protein expression and Caco-2 TJ permeability. The TNF-alpha-induced activation of MLCK promoter was mediated by binding of the activated NF-kappaB p50/p65 dimer to the downstream kappaB binding site (-84 to -75) on the MLCK promoter region; deletion of the kappaB binding site prevented the TNF-alpha increase in promoter activity. Additionally, siRNA silencing of NF-kappaB p65 also prevented the TNF alpha increase in MLCK promoter activity. In conclusion, our findings indicated that the TNF-alpha-induced increase in intestinal epithelial TJ permeability was mediated by NF-kappaB p50/p65 binding and activation of the MLCK promoter. NF kappaB p50/p65 activation of the MLCK promoter then leads to a stepwise increase in MLCK transcription, expression and activity, and MLCK-mediated opening of the intestinal TJ barrier. PMID- 16474010 TI - Different effects of endotoxic shock on the respiratory function of liver and heart mitochondria in rats. AB - This study was designed to clarify whether mitochondrial function/dysfunction and reactive oxygen species (ROS) production have a temporal relationship with organ failure during endotoxic shock. Adult male Sprague-Dawley rats were divided into three groups receiving 1) isotonic saline (control group, n = 16); 2) 8 mg/kg lipopolysaccharide (LPS; n = 8); or 3) 20 mg/kg LPS (n = 8) intraperitoneally under short anesthesia with 3.5% of isoflurane. After 16 h, animals were killed to analyze plasma, rat liver mitochondria (RLM), and rat heart mitochondria (RHM). In accordance with plasma analysis, LPS-treated rats were divided into "responders" and "nonresponders" with high and low levels of alanine aminotransferase and creatine, respectively. RHM from responders had significantly lower respiratory activity in state 3, suggesting a decreased rate of ATP synthesis. In contrast, RLM from responders had significantly higher respiratory activity in state 3 than both nonresponders and the control group. This increase was accompanied by a decrease in phosphate-to-oxygen ratio values, which was not observed in RHM. ROS generation determined with a spin probe, 1 hydroxy-3-carboxypyrrolidine, neither revealed a difference in RHM between LPS and control groups nor between responders and nonresponders. In contrast, RLM isolated from responders showed a marked increase in ROS production compared with both the control group and nonresponders. Our data demonstrate that 1) RHM and RLM respond to endotoxic shock in a different manner, decreasing and increasing respiratory activity, respectively, and 2) there is a temporal relationship between ROS production in RLM (but not in RHM) and tissue damage in rats subjected to LPS shock. PMID- 16474011 TI - Vectorial transport of unconjugated and conjugated bile salts by monolayers of LLC-PK1 cells doubly transfected with human NTCP and BSEP or with rat Ntcp and Bsep. AB - Na(+)-taurocholate-cotransporting peptide (NTCP)/SLC10A1 and bile salt export pump (BSEP)/ABCB11 synergistically play an important role in the transport of bile salts by the hepatocyte. In this study, we transfected human NTCP and BSEP or rat Ntcp and Bsep into LLC-PK1 cells, a cell line devoid of bile salts transporters. Transport by these cells was characterized with a focus on substrate specificity between rats and humans. The basal to apical flux of taurocholate across NTCP- and BSEP-expressing LLC-PK1 monolayers was 10 times higher than that in the opposite direction, whereas the flux across the monolayer of control and NTCP or BSEP single-expressing cells did not show any vectorial transport. The basal to apical flux of taurocholate was saturated with a K(m) value of 20 microM. Vectorial transcellular transport was also observed for cholate, chenodeoxycholate, ursodeoxycholate, their taurine and glycine conjugates, and taurodeoxycholate and glycodeoxycholate, whereas no transport of lithocholate was detected. To evaluate the respective functions of NTCP and BSEP and to compare them with those of rat Ntcp and Bsep, we calculated the clearance by each transporter in this system. A good correlation in the clearance of the examined bile salts (cholate, chenodeoxycholate, ursodeoxycholate, and their taurine or glycine conjugates) was observed between transport by human and that of rat transporters in terms of their rank order: for NTCP, taurine conjugates > glycine conjugates > unconjugated bile salts, and for BSEP, unconjugated bile salts and glycine conjugates > taurine conjugates. In conclusion, the substrate specificity of human and rat NTCP and BSEP appear to be very similar at least for monovalent bile salts under physiological conditions. PMID- 16474012 TI - Infertility caused by polymorphisms or mutations in spermatogenesis-specific genes. PMID- 16474013 TI - Feedback inhibition of gonadotropins by testosterone in men with hypogonadotropic hypogonadism: comparison to the intact pituitary-testicular axis in primary hypogonadism. AB - Men with hypogonadotropic hypogonadism (HH) due to hypothalamic-pituitary disease present with low serum testosterone levels combined with undetectable, low, or normal gonadotropin levels. Treatment consists of testosterone replacement to reverse the symptoms of androgen deficiency. The aim of this study was to examine the dynamics and feedback inhibition of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in relation to testosterone in 38 men with HH treated with testosterone. Findings were compared with 11 men with primary hypergonadism (PH). Testosterone replacement led to a suppression of FSH levels from 2.8 IU/L at baseline to 1.1 IU/L and to a suppression of LH levels from 2.3 to 0.8 IU/L. There was a linear correlation between levels of FSH and LH (after natural log transformation for both) and testosterone levels in both the HH and PH groups. However, the differences in intercepts and slopes between the groups were significant. To determine whether nonsuppressed FSH or LH during testosterone replacement reduces the probability of eugonadism, as reflected by normal testosterone levels, gonadotropin levels were measured and categorized as low (<0.5 IU/L), medium (0.5-2 IU/L), and high levels (>2 IU/L). The higher FSH or LH levels were found to significantly decrease the chance for achieving eugonadism. In conclusion, in men with HH due to hypothalamic-pituitary disease or injury, the pituitary-testicular hormonal axis maintains its physiological negative feedback between testosterone and gonadotropins. Thus, gonadotropin levels in men with HH might be useful, together with testosterone concentrations, for assessing the adequacy of androgen replacement. PMID- 16474014 TI - A massive snowball of fraud and deceit. PMID- 16474016 TI - Growth factors and the epididymis. PMID- 16474017 TI - Sperm nuclear histone to protamine ratio in fertile and infertile men: evidence of heterogeneous subpopulations of spermatozoa in the ejaculate. AB - Sperm protamine deficiency is observed in a subset of infertile men, suggesting that the relative histone to protamine ratio may be altered in the spermatozoa of these men. We measured the ratio of nuclear histones to protamines in the spermatozoa of fertile (n = 10) and infertile men (n = 20). Sperm nuclear proteins were extracted and subsequently separated by acid-urea (AU) polyacrylamide gel electrophoresis. The relative histone (H2B) to protamine (PRM1 + PRM2) and PRM1 to PRM2 ratios were estimated by densitometric analysis of the AU gels. Immunoblotting experiments (using H2B, PRM1, and PRM2 antibodies) were conducted to confirm the specificity of the bands. The pattern and intensity of H2B staining in human spermatozoa was assessed by immunocytochemistry. Sperm samples from the infertile men in this study had a significantly higher proportion of histone H2B to protamine (PRM1 + PRM2) than did samples from the fertile men in this study (0.38 vs 0.08, P < .001). Immunocytochemistry experiments demonstrated a punctuated staining pattern (with strong, intermediate, or weak H2B staining intensity) throughout the sperm head. Infertile men had a higher proportion of spermatozoa exhibiting strong and intermediate staining than did samples from fertile men. These findings suggest that infertile men possess a higher proportion of spermatozoa with an increased histone to protamine ratio than fertile controls. PMID- 16474018 TI - Testosterone replacement therapy for older men. PMID- 16474019 TI - Testosterone replacement therapy for male aging: ASA position statement. PMID- 16474020 TI - Investigation, treatment, and monitoring of late-onset hypogonadism in males: ISA, ISSAM, and EAU recommendations. PMID- 16474021 TI - The intelligent couch potato. PMID- 16474022 TI - Antisperm immunity for contraception. PMID- 16474023 TI - The effect of calcium channel blockers on male reproductive potential. PMID- 16474024 TI - The making of a microsurgeon. PMID- 16474025 TI - Syncope in a transsexual male. PMID- 16474026 TI - Testicular torsion alters the presence of specific proteins in the mouse testis as well as the phosphorylation status of specific proteins. AB - Testicular torsion followed by torsion repair induces an ischemia-reperfusion injury to the testis that can render the testis aspermatogenic. Previous results have demonstrated this loss of spermatogenesis to be the result of germ cell apoptosis induced by oxidative stress. The present work reports protein changes occurring in the mouse testis 24 hours after repair of a testicular torsion known to induce germ cell apoptosis and severe seminiferous impairment. Total proteins were extracted from sham-operated testes and testes having had 2-hour 720 degrees torsion 24 hours previously. Testicular proteins were separated by 2-dimensional electrophoresis and the resulting gel images were analyzed with image analysis software. Of the over 1100 proteins detected on the average gel, over 700 were consistently appearing in multiple gels, and those protein spot intensities were averaged within sham and torsion groups and compared between the 2 groups. Twenty three proteins were consistently increased after torsion repair and 48 were decreased. Six proteins, 3 of which increased and 3 of which decreased after torsion repair, were identified by mass spectrometry. The 3 proteins that increased after torsion repair, beta2-tubulin and 2 isoforms of serum albumin, as well as the 3 proteins that decreased after torsion repair, vimentin, phosphoglycerate kinase, and t-complex protein 1beta, were for the most part associated with various aspects of cell stress responses. The number of proteins phosphorylated on tyrosine residues exceeded the number of proteins phosphorylated on serine/threonine residues, but among 6 stress-related proteins specifically examined for phosphorylation in sham testes and those examined after torsion repair, increases in threonine phosphorylation of c-Jun NH2 terminal kinase and activating transcription factor 2 were the most prominent. Knowing these proteins and the pathways to which they point will aid in the search for new therapies of oxidative stress in the testis. PMID- 16474027 TI - Double-strand break repair in plants is developmentally regulated. AB - In this study, we analyzed double-strand break (DSB) repair in Arabidopsis (Arabidopsis thaliana) at various developmental stages. To analyze DSB repair, we used a homologous recombination (HR) and point mutation reversion assays based on nonfunctional beta-glucuronidase reporter genes. Activation of the reporter gene through HR or point mutation reversion resulted in the appearance of blue sectors after histochemical staining. Scoring of these sectors at 3-d intervals from 2 to 31 d post germination (dpg) revealed that, although there was a 100-fold increase in the number of genomes per plant, the recombination frequency only increased 30 fold. This translates to a recombination rate at 31 dpg (2.77 x 10(-8)) being only 30% of the recombination rate at 2 dpg (9.14 x 10(-8)). Conversely, the mutation frequency increased nearly 180-fold, resulting in a 1.8-fold increase in mutation rate from 2 to 31 dpg. Additional analysis of DSBs over the early developmental stages revealed a substantial increase in the number of strand breaks per unit of DNA. Furthermore, RNA analysis of Ku70 and Rad51, two key enzymes in two different DSB repair pathways, and further protein analysis of Ku70 revealed an increase in Ku70 levels and a decrease of Rad51 levels in the developing plants. These data suggest that DSB repair mechanisms are developmentally regulated in Arabidopsis, whereby the proportion of breaks repaired via HR substantially decreases as the plants mature. PMID- 16474028 TI - Weekly rifapentine/isoniazid or daily rifampin/pyrazinamide for latent tuberculosis in household contacts. AB - RATIONALE: Treatment of latent tuberculosis (TB) infection with weekly rifapentine and isoniazid is a potentially effective alternative to current therapies. OBJECTIVES: To compare the efficacy of weekly rifapentine/isoniazid to daily rifampin/pyrazinamide in preventing TB in household contacts of patients with pulmonary TB in Brazil. METHODS: Contacts of patients with TB were randomized to rifapentine 900 mg/isoniazid 900 mg once weekly for 12 wk or rifampin 450-600 mg/pyrazinamide 750-1,500 mg daily for 8 wk and followed for at least 2 yr. MEASUREMENTS: TB rates, adverse events, and adherence to therapy. MAIN RESULTS: A total of 399 household contacts were enrolled, 206 in the rifapentine/isoniazid arm and 193 in the rifampin/pyrazinamide arm. The median age was 34 yr, median weight was 63 kg, 60% of participants were female, and only one patient was HIV infected. Rifapentine/isoniazid was well tolerated, but the trial was halted by the investigators before completion because of unanticipated hepatotoxicity in the rifampin/pyrazinamide arm. Twenty of 193 participants (10%) receiving rifampin/pyrazinamide experienced grade 3 or 4 hepatotoxicity, compared with 2 of 206 participants (1%) on rifapentine/isoniazid (p<0.001). There were no hospitalizations or deaths due to hepatotoxicity, and all participants' liver enzyme levels returned to normal during follow-up. During follow-up, four cases of active TB developed, three in the rifapentine/isoniazid group and one in the rifampin/pyrazinamide group (1.46 vs. 0.52%; difference, 0.94%; 95% confidence interval, -1.6 to 3.7%). CONCLUSIONS: Rifapentine/isoniazid was better tolerated than rifampin/pyrazinamide and was associated with good protection against TB. Rifapentine/isoniazid weekly for 12 wk is likely a promising therapy for latent TB infection. PMID- 16474029 TI - Postoperative pneumonia after major lung resection. AB - BACKGROUND: Postoperative pneumonia (POP) is a life-threatening complication of lung resection. The incidence, causative bacteria, predisposing factors, and outcome are poorly understood. DESIGN: Prospective observational study. METHODS: A prospective study of all patients undergoing major lung resections for noninfectious disease was performed over a 6-mo period. Culture of intraoperative bronchial aspirates was systematically performed. All patients with suspicion of pneumonia underwent bronchoscopic sampling and culture before antibiotherapy. RESULTS: One hundred and sixty-eight patients were included in the study. Bronchial colonization was identified in 31 of 136 patients (22.8%) on analysis of intraoperative samples. The incidence of POP was 25% (42 of 168). Microbiologically documented and nondocumented pneumonias were recorded in 24 and 18 cases, respectively. Haemophilus species, Streptococcus species, and, to a much lesser extent, Pseudomonas and Serratia species were the most frequently identified pathogens. Among colonized and noncolonized patients, POP occurred in 15 of 31 and 20 of 105 cases, respectively (p = 0.0010; relative risk, 2.54). Death occurred in 8 of 42 patients who developed POP and in 3 of 126 of patients who did not (p = 0.0012). Patients with POP required noninvasive ventilation or reintubation more frequently than patients who did not develop POP (p < 0.0000001 and p = 0.00075, respectively). POP was associated with longer intensive care unit and hospital stay (p < 0.0000001 and p = 0.0000005, respectively). Multivariate analysis showed that chronic obstructive pulmonary disease, extent of resection, presence of intraoperative bronchial colonization, and male sex were independent risk factors for POP. CONCLUSIONS: Pneumonia acquired in hospital represents a relatively frequent complication of lung resections, associated with an important percentage of postoperative morbidity and mortality. PMID- 16474030 TI - Airway inflammation and bronchial bacterial colonization in chronic obstructive pulmonary disease. AB - RATIONALE: Inflammation is now recognized as an integral part of the pathogenesis of chronic obstructive pulmonary disease (COPD). In contrast to the sterile airways of normal lungs, bacterial pathogens are often isolated from the airways in stable COPD. This "colonization" of the tracheobronchial tree, currently believed to be innocuous, could serve as an inflammatory stimulus, independent of current tobacco smoke exposure. OBJECTIVE: To test the hypothesis that bacterial colonization is associated with airway inflammation in stable COPD. METHODS: Bronchoscopy with bronchoalveolar lavage (BAL) was performed in three groups of subjects: 26 ex-smokers with stable COPD (COPD), 20 ex-smokers without COPD (ex smokers), and 15 healthy nonsmokers (nonsmokers). Quantitative bacterial cultures, cell counts, chemokine, cytokine, proteinase/antiproteinase, and endotoxin levels in the BAL fluid were compared. RESULTS: Potentially pathogenic bacteria were recovered at > or = 100 cfu/ml in 34.6% of COPD, 0% of ex-smokers, and in 6.7% of nonsmokers (p = 0.003). All values are expressed as median (interquartile range). Subjects with colonized COPD had significantly greater relative (12.0 [28.4] vs. 3.0 [7.8]%, p = 0.03) and absolute (4.98 [5.26] x 10(4)/ml vs. 3.04 [2.82] x 10(4)/ml, p = 0.02) neutrophil counts, interleukin 8 (33.8 [189.8] vs. 16.9 [20.1] pg/ml, p = 0.005), active matrix metalloproteinase 9 (2.16 [4.30] vs. 0.84 [0.99] U/ml, p = 0.03), and endotoxin (36.0 [72.6] vs. 3.55 [7.17] mEU/ml, p = 0.004) levels in the BAL than the subjects with noncolonized COPD. These inflammatory constituents of BAL were also significantly elevated in subjects with colonized COPD when compared with ex-smokers and nonsmokers. CONCLUSIONS: Bacterial colonization is associated with neutrophilic airway lumen inflammation in ex-smokers with COPD and could contribute to progression of airway disease in COPD. PMID- 16474031 TI - Persistent infection of Chlamydia in reactive arthritis. PMID- 16474033 TI - Antiphospholipid antibody syndrome associated with primary angiitis of the central nervous system: report of two biopsy proven cases. PMID- 16474032 TI - Anti-cyclic citrullinated peptide antibodies in hepatitis C virus associated rheumatological manifestations and Sjogren's syndrome. AB - OBJECTIVE: To investigate the diagnostic reliability of anti-CCP antibodies (anti CCP Ab) in distinguishing hepatitis C virus (HCV) associated rheumatological manifestations and Sjogren's syndrome from rheumatoid arthritis. METHODS: 147 HCV infected patients (HCV RNA positive) were compared with 64 patients with definite rheumatoid arthritis in a retrospective study. Anti-CCP Ab were detected using the Immunoscan ELISA kit (second generation) and rheumatoid factor (RF) by the FIDIStrade mark Rheuma kit. RESULTS: Among the 147 HCV infected patients (77 women; mean (SD) age 58 (16) years), 77 (52%) had a mixed cryoglobulin (MC), 38 (26%) an MC associated systemic vasculitis, 35 (24%) arthralgia/arthritis, and seven (5%) definite Sjogren's syndrome. HCV infected patients with arthralgia were more often RF positive than those without arthralgia (54% v 27%; p = 0.003), but less often than patients with rheumatoid arthritis (54% v 81%; p = 0.009). Anti-CCP Ab were detected in only two HCV infected patients with arthralgia (5.7%), in none without arthralgia or with Sjogren's syndrome, and in 78% of patients with rheumatoid arthritis. With a specificity of 93.5% and a positive predictive value of 96% for rheumatoid arthritis, anti-CCP Ab were the most specific biological marker. CONCLUSIONS: Anti-CCP antibodies are very rarely found in HCV infected patients with rheumatological manifestations or Sjogren's syndrome. They are reliable serological markers to distinguish these from patients with rheumatoid arthritis. PMID- 16474034 TI - ANCA associated glomerulonephritis in a patient with mixed connective tissue disease. PMID- 16474035 TI - Three dimensional power Doppler ultrasonography confirms early reduction of synovial perfusion after intra-articular steroid injection. PMID- 16474036 TI - Alveolar and serum antiphospholipid antibodies in acute respiratory distress syndrome associated with catastrophic antiphospholipid syndrome. PMID- 16474037 TI - Dupuytren's contracture is associated with sprouting of substance P positive nerve fibres and infiltration by mast cells. PMID- 16474038 TI - Arterial stiffness in Behcet's disease: increased regional pulse wave velocity values. PMID- 16474039 TI - Safety and efficacy of leflunomide in the treatment of lupus nephritis refractory or intolerant to traditional immunosuppressive therapy: an open label trial. PMID- 16474040 TI - No evidence of parvovirus B19 in tissue samples from patients with polyarteritis nodosa and microscopic polyangiitis as assessed by the polymerase chain reaction. PMID- 16474042 TI - Viral meningitis. AB - Viruses probably account for most cases of acute meningitis. Viral meningitis is often assumed to be a largely benign disease. For the commonest pathogens causing meningitis, enteroviruses, this is usually the case; however, for many of the other pathogens causing viral meningitis, and for common pathogens in the immunocompromised or infants, viral meningitis is frequently associated with substantial neurological complications and a significant mortality. Diagnostic methods for rapid and accurate identification of pathogens have improved over recent years, permitting more precise and earlier diagnoses. There have been fewer developments in therapies for viral meningitis, and there remain no effective therapies for most pathogens, emphasising the importance of prevention and early diagnosis. This review focuses on the presentation, diagnosis and management of viral meningitis and also covers the prevention of meningitis for pathogens where effective vaccines are available. PMID- 16474045 TI - Judgement postponed in disciplinary case. PMID- 16474043 TI - Living donor liver transplantation: present status and recent advances. AB - The first successful living donor liver transplantation (LDLT) was performed in a child in 1989 in Brisbane and in an adult in 1994 by the Shinshu group. Over the past few years, LDLT has increased worldwide and is now an established alternative to deceased donor liver transplantation. The surgical procedures for LDLT are more technically challenging than those for whole liver transplantation. LDLT requires a full understanding of the hepatobiliary anatomy and continuous technical refinement of the procedure. Some of the technical highlights include selective vascular occlusion techniques for donor hepatectomy, hepatic arterial reconstruction under the microscope and the introduction of intraoperative ultrasound, graft volume estimation and hepatic venous reconstruction, all of which have improved the success rate of LDLT over the past few years. This review focuses on recent trends and surgical techniques for LDLT. PMID- 16474051 TI - Description of the outbreak of equine influenza (H3N8) in the United Kingdom in 2003, during which recently vaccinated horses in Newmarket developed respiratory disease. AB - Between March and May 2003, equine influenza virus infection was confirmed as the cause of clinical respiratory disease among both vaccinated and unvaccinated horses of different breeds and types in at least 12 locations in the UK. In the largest outbreak, 21 thoroughbred training yards in Newmarket, with more than 1300 racehorses, were affected, with the horses showing signs of coughing and nasal discharge during a period of nine weeks. Many of the infected horses had been vaccinated during the previous three months with a vaccine that contained representatives from both the European (A/eq/Newmarket/2/93) and American (A/eq/Newmarket/1/93) H3NN8 influenza virus lineages. Antigenic and genetic characterisation of the viruses from Newmarket and elsewhere indicated that they were all closely related to representatives of a sublineage of American viruses, for example, Kentucky/5/02, the first time that this sublineage had been isolated in the uk. In the recently vaccinated racehorses in Newmarket the single radial haemolysis antibody levels in acute sera appeared to be adequate, and there did not appear to be significant antigenic differences between the infecting virus and A/eq/Newmarket/1/93, the representative of the American lineage virus present in the most widely used vaccine, to explain the vaccine failure. However, there was evidence for significantly fewer infections among two-year-old horses than older animals, despite their having similar high levels of antibody, consistent with a qualitative rather than a quantitative difference in the immunity conveyed by the vaccination. PMID- 16474053 TI - Measurements of growth hormone and insulin-like growth factor 1 in cats with diabetes mellitus. AB - Serum concentrations of insulin-like growth factor 1 (IGF-1) and growth hormone were measured in 25 cats with untreated diabetes mellitus (11 of which were used for follow-up measurements, one to three, four to eight, nine to 12 and 13 to 16 weeks after their treatment with insulin began), 14 diabetic cats that had previously been treated with insulin, and seven diabetic cats that also had hypersomatotropism, two of which had not previously been treated with insulin; 18 healthy cats were used as controls. In the untreated diabetic cats the concentration of IGF-1 ranged from 13.0 to 433.0 ng/ml (median 170.5 ng/ml), which was significantly lower than the concentrations in the control cats (196.0 to 791.0 ng/ml, median 452.0 ng/ml). Their IGF-1 concentrations increased significantly when they were treated with insulin and after four to eight weeks were not different from those in the control cats. In the diabetic cats that had previously been treated with insulin the IGF-1 concentrations were 33.0 to 476.0 ng/ml (median 316.0 ng/ml), which was significantly lower than the concentrations in the control cats, but significantly higher than in the untreated diabetic cats. The IGF-1 concentrations in the two previously untreated diabetic cats with hypersomatotropism were low and low-normal but increased markedly after treatment with insulin. In the five previously treated cats with hypersomatotropism the concentration of IGF-1 was above the normal range. The concentrations of growth hormone in the treated and untreated diabetic cats without hypersomatotropisms were not significantly different and there was an overlap in its concentrations in the diabetic cats with and without hypersomatotropism. PMID- 16474054 TI - Lack of susceptibility of the dromedary camel (Camelus dromedarius) to foot-and mouth disease virus serotype O. PMID- 16474052 TI - Comparison of an antifungal agent with a mixture of antifungal, antibiotic and corticosteroid agents for the treatment of Malassezia species otitis in dogs. AB - Twenty dogs with otitis externa in both ears and numerous Malassezia species yeasts on cytological examination were treated in one ear with a combination product containing clotrimazole, marbofloxacin and dexamethasone, and in the other ear with a topical antifungal containing miconazole. The effects of the treatments were analysed on the basis of the scores for pruritus, erythema and amount of cerumen, and the number of yeasts on cytological smears. There were reductions in the counts of Malassezia species after both treatments, but the combination product gave significantly greater reductions in erythema, cerumen and pruritus. PMID- 16474055 TI - Unusual mortality incidents in tit species (family Paridae) associated with the novel bacterium Suttonella ornithocola. PMID- 16474056 TI - Halitosis in association with canine hepatozoonosis. PMID- 16474057 TI - Evaluation of an all-glass impinger for the detection of porcine reproductive and respiratory syndrome virus in natural and artificial aerosols. PMID- 16474059 TI - Humane slaughter of birds. PMID- 16474060 TI - Humane slaughter of birds. PMID- 16474062 TI - 'Normal' serum chemistry values in wild animals. PMID- 16474063 TI - Non-therapeutic docking of dogs' tails. PMID- 16474064 TI - Non-therapeutic docking of dogs' tails. PMID- 16474065 TI - Veterinary acupuncture. PMID- 16474066 TI - Lymphoid leukaemia in a saker falcon. PMID- 16474067 TI - Measuring malaria. PMID- 16474068 TI - Comparing methods of estimating the global morbidity burden from Plasmodium falciparum malaria. PMID- 16474069 TI - High titers of IgG antibodies against Plasmodium falciparum serine repeat antigen 5 (SERA5) are associated with protection against severe malaria in Ugandan children. AB - Plasmodium falciparum serine repeat antigen (SERA5) is a promising asexual blood stage malaria candidate vaccine. However, there is a paucity of information about natural immune responses to SERA5 in children from malaria-endemic regions. We undertook a hospital-based case-control study of severe malaria in Apac District, Northern Uganda, in children 6-59 months of age. The commonest symptoms observed in children with severe malaria (SM) were respiratory distress (53.4%) and prostration (40.4%) followed by circulatory collapse (7.4%), severe anemia (Hb < 5 g/dL, 7.0%), and seizures (2.6%). None of the SM children had impaired consciousness, coma, or cerebral malaria. We measured serum IgG antibodies using a recombinant construct of SERA5 (SE36) in enzyme-linked immunosorbent assays. High titers of IgG anti-SE36 were associated with protection against severe malaria in children under 5 years old. PMID- 16474070 TI - Point mutations in the dihydrofolate reductase and dihydropteroate synthase genes of Plasmodium falciparum and resistance to sulfadoxine-pyrimethamine in Sri Lanka. AB - Sulfadoxine-pyrimethamine (SP) is the second-line treatment for Plasmodium falciparum malaria in Sri Lanka. Resistance to SP is caused by point mutations in the dihydrofolate reductase (Pf-dhfr) and dihydropteroate synthase (Pf-dhps) genes of P. falciparum. We determined the genotype of Pf-dhfr and Pf-dhps and the clinical response to SP in 30 field isolates of P. falciparum from Sri Lanka. All patients treated with SP had an adequate clinical response. Eighty-five percent (23 of 27) of pure field isolates carried parasites with double mutant alleles of Pf-dhfr (C59R + S108N) and showed about 200-fold higher levels of resistance to pyrimethamine than the wild type in a yeast system. None of the isolates had either known or novel mutations at other positions in the dhfr domain. In contrast, 67% (20 of 30) of the isolates carried parasites that were wild type for Pf-dhps. In Sri Lanka, detection of the triple mutant allele of Pf-dhfr will require tracking mutations at codon 51. PMID- 16474071 TI - Frequency distribution of antimalarial drug-resistant alleles among isolates of Plasmodium falciparum in Bangui, Central African Republic. AB - We determined the baseline frequency distribution of mutant alleles of genes associated with resistance to chloroquine and sulfadoxine-pyrimethamine in Plasmodium falciparum isolates in Bangui, Central African Republic. Mutant alleles of the P. falciparum chloroquine resistance transporter (pfcrt) gene were found in all samples and the frequency of the deduced CIET pfcrt haplotype was high (45%). The most common allele of the P. falciparum multidrug resistance 1 (pfmdr1) gene among the field isolates of P. falciparum was 86Y (21.9%). The 1246Y allele was also common (18.0%). Of the 167 P. falciparum isolates in which the dihydrofolate reductase gene was studied, only 11 carried the wild-type allele (6.6%) whereas many (50.3%) were quadruple mutants (50R, 51I, 59R, 108N). The frequency of the 436A mutant allele of the dihydropteroate synthase gene was high (74.3%), but the frequencies of the 437G (18.6%) and 540E (5.2%) mutant alleles were low. Molecular analyses of antimalarial drug-resistant alleles of P. falciparum isolates in Bangui strongly suggest the widespread distribution of chloroquine and pyrimethamine resistance and to a lesser extent sulfadoxine resistance. PMID- 16474072 TI - A case-control auditory evaluation of patients treated with artemether lumefantrine. AB - Artemether-lumefantrine is the first registered, fixed, artemisinin-based combination treatment. Artemisinin derivatives are highly effective antimalarials with a favorable safety profile. Concerns remain over their potential neurotoxicity, although there has been no clinical evidence of this in humans. In animals (rats, dogs, and monkeys) artemether, a derivative of artemisinin is associated with an unusual toxicity pattern in specific brain nuclei involving the auditory and vestibular pathways. A recent report from Mozambique described a small but significant and irreversible hearing loss in patients exposed to artemether-lumefantrine. To explore this issue, we conducted a case-control study using tympanometry, audiometry and auditory brain-stem responses. We assessed 68 subjects who had been treated with artemether-lumefantrine within the previous five years and 68 age- and sex-matched controls living in the malarious region along the Thailand-Myanmar border. There were no differences in the test results between cases and controls. There was no neurophysiologic evidence of auditory brainstem toxicity that could be attributed to artemether-lumefantrine in this study population. PMID- 16474073 TI - Assessment of transmission-blocking activity of candidate Pvs25 vaccine using gametocytes from chimpanzees. AB - Macaca mulatta monkeys were immunized with the candidate transmission-blocking vaccine against Plasmodium vivax, Pvs25, combined with alum or Montanide ISA 720. Efficacy was measured by combining post-immunization sera with gametocytes obtained from infections induced in chimpanzees using membrane-feeding techniques. The results indicate that immunization of M. mulatta monkeys with Pvs25 and Montanide ISA 720 was more effective than with alum in efficacy and resulted in the maintenance of a lasting transmission-blocking immunity to P. vivax. This was evident two weeks after the second immunization, and more strongly demonstrable 62 and 152 days after the second immunization. This transmission-blocking activity was strongly reinforced by a third immunization given 181 days after the primary immunization, as measured three weeks later by indirect membrane feeding. The use of gametocytes of P. vivax derived from infections induced in chimpanzees can contribute to the selection of appropriate constructs, formulations, and immunization regimens for the development of effective transmission-blocking vaccines. PMID- 16474074 TI - Antibodies to Bartonella vinsonii subsp. Berkhoffii in Moroccan dogs. AB - Bartonella vinsonii subsp. berkhoffii is a fastidious microorganism that has been recognized as an emerging human and canine pathogen. We report for the first time on the prevalence of antibodies to B. vinsonii subsp. berkhoffii in domestic dogs from Morocco. The overall seroprevalence was 38% (56 of 147 dogs tested). Most of the seropositive dogs were stray dogs from Rabat (36%, 8 of 22) and Khenifra (47%, 47 of 101). Antibodies against B. vinsonii subsp. berkhoffii were found infrequently among pet dogs from Rabat (4%, 1 of 24). PMID- 16474075 TI - Quantitative analysis of La Crosse virus transcription and replication in cell cultures and mosquitoes. AB - La Crosse (LAC) virus (family Bunyaviridae, genus Orthobunyavirus) small (S) segment negative-sense RNA genome (vRNA), positive-sense full-length RNA complement (vcRNA), and subgenomic mRNA were assayed in infected cell cultures and female Aedes (Ochlerotatus) triseriatus mosquito tissues using quantitative PCR (Q-PCR). During persistent infection of C6/36 (Aedes albopictus) and MAT (Aedes triseriatus) cultured cells and cytolytic infection of BHK-21 cultured cells, LAC vRNA was the most abundant RNA species, followed by mRNA and vcRNA. RNA copy numbers per cell were quantified and vRNA correlated to virus titer in cell culture medium. The Q-PCR assay proved more sensitive than reverse transcription (RT)-PCR and immunofluorescence assays (IFA) for detecting LAC virus infection of mosquitoes. After infection of female mosquitoes orally, quantities of LAC RNA increased in ovaries for 6 days, and as ovarian biosynthetic activity quiesced, LAC RNA quantities decreased then remained detectable at a low level. After a second, noninfectious blood meal, quantities of LAC RNA in ovaries increased significantly, quantitatively confirming correlation of LAC virus RNA synthesis with vector metabolic activity. Coregulation of viral replication and mosquito ovary metabolic activity may condition efficient transovarial transmission. PMID- 16474076 TI - Prevalence of HIV and other sexually transmitted infections among female commercial sex workers in Argentina. AB - Sexually transmitted infections (STIs) have shown to enhance the transmission of human immunodeficiency virus (HIV) and to be more common among female commercial sex workers (FSWs). A cross-sectional study was conducted among 625 FSWs in six cities of Argentina in 2000-2002. The seroprevalence of HIV, hepatitis B virus (HBV), hepatitis C virus (HCV), human T-cell lymphotropic virus type I/II, and syphilis was 3.2%, 14.4%, 4.3%, 1.6%, and 45.7%, respectively. Syphilis was associated with older age (>/= 30 years, adjusted odds ratio [AOR] = 2.6 to 4.9), >/= 10 years in sex work (AOR = 2.2), use of illegal drugs (AOR = 2.1), and a prior history of an STI (AOR = 3.0). HBV and syphilis was the most common co infection in 44 (7.5%) subjects. FSWs in Argentina are exposed to HIV and other STIs due to high-risk sexual and illegal drug use behavior. Renewed efforts are necessary to intervene effectively in this high-risk population. PMID- 16474077 TI - Morbidity and mortality profile of human immunodeficiency virus-infected patients with and without hepatitis C co-infection. AB - Hepatitis C virus (HCV) and human immunodeficiency virus (HIV) co-infection is an important and frequent scenario, predominantly in injecting drug users (IDUs). The present study evaluated morbidity and mortality variation in HIV-infected patients with and without HCV co-infection. Co-infection prevalence was determined in 356 HIV-infected persons. Their clinical manifestations, laboratory findings, risk factors, HIV therapies, and mortality rates were evaluated. The prevalence of HCV was 54% in the overall group and 81% in IDUs, with a predominance of HCV genotype 1. Mortality rates were similar in patients with and without co-infection; however, co-infected patients had significantly higher liver damage as a cause of mortality when compared with those who were not co infected. The high prevalence of HCV and an emerging mortality from liver diseases showed the significance of this co-infection in the HIV epidemic. Primary and secondary prevention are necessary to reduce the expanding impact of HCV infection in HIV patients. PMID- 16474078 TI - Impairment of host immune response against strongyloides stercoralis by human T cell lymphotropic virus type 1 infection. AB - A large-scale study was undertaken to clarify the prevalence rate of strongyloidiasis in Okinawa, Japan and to evaluate the relationship between strongyloidiasis and infection with human T cell lymphotropic virus type 1 (HTLV 1). The prevalence rate of Strongyloides stercoralis and HTLV-1 infection were 6.3% and 14.0%, respectively. Among 2,185 patients more than 50 years of age, the rate of S. stercoralis infection was significantly higher in patients with HTLV-1 infection compared with patients without HTLV-1 infection. In 252 patients treated with ivermectin, serum IgE levels and peripheral eosinophil counts were significantly lower in HTLV-1 co-infected patients compared with patients without HTLV-1 infection. In addition, the anthelmintic effect was significantly lower in patients with HTLV-1 infection compared with patients without HTLV-1 infection. Our prospective study demonstrated a prevalence rate for strongyloidiasis and HTLV-1 infections, and clearly demonstrated that co-infection with HTLV-1 impaired the immune response against S. stercoralis. PMID- 16474080 TI - Immunogenicity of a locally produced hepatitis B vaccine with the birth dose stored outside the cold chain in rural Vietnam. AB - The heat stability of hepatitis B vaccine (HepB vaccine) should enable its storage outside the cold chain (OCC), increasing access to the birth dose in areas lacking refrigeration. We compared the immunogenicity of a locally produced vaccine among infants who received three doses stored within the cold chain (n = 358) or for whom the first dose was stored OCC for up to one month (n = 748). Serum was collected from these infants at age 9-18 months. The vaccine was protective in 80.3% of all infants. There were no differences in the prevalence of a protective level of antibody or antibody titer among groups of infants according to storage strategy. Differences in antibody titer between certain groups of infants could be explained by different vaccination schedules. Where birth dose coverage will be improved, HepB vaccine can be taken OCC for up to one month without affecting its immunogenicity. PMID- 16474079 TI - Hepatitis E virus infection as a marker for contaminated community drinking water sources in Tibetan villages. AB - In April-May 2001, a study was conducted to determine the prevalence of antibodies against hepatitis E virus (HEV) among 426 persons 8-49 years of age randomly selected from two groups of rural villages in central Tibet. Group 1 villages were assessed in 1998 as having poor quality water sources; new water systems were then constructed prior to this study. Group 2 villages had higher quality water and were not designated as priority villages for new systems prior to the study. No participants tested positive for IgM; only IgG was detected in the analyzed samples. Overall, 31% of the participants had ever been infected with HEV (95% confidence interval [CI] = 26.7-35.7%). The rate was higher in men (36.6%) than women (26.3%) and highest in those 30-39 years of age (49.1%). The rate of past infection was higher in group 1; the risk ratio was 2.77 (95% CI = 1.98-3.88). This difference is most likely the result of the poor quality of the original water sources in these villages. In resource-poor countries, HEV may be a useful health indicator reflecting the degree of contamination in village water sources. This may be especially important in rural areas (such as Tibet) where maternal mortality ratios are high because HEV may be an important cause of deaths during pregnancy in disease-endemic areas. PMID- 16474081 TI - Hepatitis C virus infection in Samoa and American Samoa. AB - Little is known about the prevalence of hepatitis C virus (HCV) in Pacific islands. In this study, serum specimens collected in 1985 and 2002 among the general populations of Samoa and American Samoa were tested for antibody to HCV by a third-generation enzyme immunoassay and a recombinant immunoblot assay. Of the 3,466 specimens tested, 8 (0.2%; 95% confidence interval = 0.07-0.4%) were positive for antibody to HCV. Prevalence did not vary by location or demographic characteristic. Thus, HCV is present in the Samoas but at a low prevalence. PMID- 16474082 TI - Origin of dengue type 3 viruses associated with the dengue outbreak in Dhaka, Bangladesh, in 2000 and 2001. AB - Dengue and dengue hemorrhagic fever re-emerged in Bangladesh in 2000 and 2001 and nearly all viruses isolated were dengue type 3. Phylogenetic analyses of the envelope genes of examples of these viruses indicated that they were most closely related to recently emerged dengue type 3 viruses from neighboring Thailand and Myanmar but distinct from those from India and Sri Lanka. Since this strain of dengue virus type 3 had not been associated with unusual patterns of disease in Thailand or Myanmar, it suggested that the outbreak in Bangladesh was due to local factors after the introduction of viruses from countries to the east rather than to the evolution of an unusually virulent strain of virus in Bangladesh. PMID- 16474083 TI - Induction of neutralizing antibodies specific to dengue virus serotypes 2 and 4 by a bivalent antigen composed of linked envelope domains III of these two serotypes. AB - There is no vaccine to prevent dengue fever, a mosquito-borne viral disease, caused by four serotypes of dengue viruses. In this study, which has been prompted by the emergence of dengue virus envelope domain III as a promising sub unit vaccine candidate, we have examined the possibility of developing a chimeric bivalent antigen with the potential to elicit neutralizing antibodies against two serotypes simultaneously. We created a chimeric dengue antigen by splicing envelope domain IIIs of serotypes 2 and 4. It was expressed in Escherichia coli and purified to near homogeneity. This protein retains the antigenic identities of both its precursors. It elicited antibodies that could efficiently block host cell binding of both serotypes 2 and 4 of dengue virus and neutralize their infectivity (neutralizing antibody titers approximately 1:40 and ~1:80 for dengue virus serotypes 2 and 4, respectively). This work could be a forerunner to the development of a single envelope domain III-based tetravalent antigen. PMID- 16474084 TI - Seroprevalence of leptospirosis among high-risk population of Andaman Islands, India. AB - Leptospirosis is a severe spirochetal zoonosis in the world. It is considered an occupational disease of persons engaged in agriculture, sewage works, forestry, and animal slaughtering. A study was conducted with an objective of assessing the seroprevalence of leptospirosis among the high-risk groups of Andaman Islands. A total of 611 sera samples from different high-risk populations were collected and tested by microscopic agglutination test (MAT). Genetic characterization of the isolate was done by randomly amplified polymorphic DNA (RAPD) fingerprinting, and serological characterization was done using monoclonal antibody technique. Antibodies to leptospires were detected in 322 samples giving an overall seroprevalence of 52.7%. The seroprevalence was highest among agriculture workers (62.5%) followed by sewage workers (39.4%), animal handlers (37.5%), forest workers (27.3%), and butchers (30.0%). Seroprevalence among control population was 14.7%, which was comparatively less than that of the high-risk population groups. Subject sera were most commonly reacted with organisms of the serogroup Grippotyphosa followed by Australis, and the pattern was similar in control group. Four leptospires were isolated from agriculture workers who were admitted to the public health center (PHC) with complaints of fever and body ache. Human isolates were compared with two rodent (Rattus norvegicus) isolates from the same area of agriculture workers to get initial information about the transmission cycle of leptospirosis in the study community. Randomly amplified polymorphic DNA fingerprinting pattern of the strains recovered from the rodents and human patients were identified as belonged to genomo-species Leptospira interrogans. The antigenic characterization of the strains recovered from them belonged to serovar Valbuzzi of serogroup Grippotyphosa. The study showed that people engaged in high-risk activities such as agriculture, sewage cleaning, animal handling, animal slaughtering, and forestry are frequently exposed to leptospirosis, and hence control strategies targeting these populations could be more effective. PMID- 16474085 TI - Pathways of expansion and multiple introductions illustrated by large genetic differentiation among worldwide populations of the southern house mosquito. AB - The southern house mosquito Culex quinquefasciatus is a principal vector of human lymphatic filariasis, several encephalitides (including West Nile virus), avian malaria, and poxvirus, but its importance as a vector varies considerably among regions. This species has spread with humans and is ubiquitous in tropical urban and suburban environments. This was the first mosquito to reach Hawaii and we performed a worldwide genetic survey using micro-satellite loci to identify its source. Our analyses showed divergent Old World and New World genetic signatures in Cx. quinquefasciatus with further distinctions between east and west African, Asian, and Pacific populations that correlate with the epidemiology of human filariasis. We found that in Hawaii south Pacific mosquitoes have largely replaced the original New World introduction of Cx. quinquefasciatus, consistent with their reported expansion to higher elevations. We hypothesize worldwide pathways of expansion of this disease vector. PMID- 16474086 TI - Use of the pupal survey technique for measuring Aedes aegypti (Diptera: Culicidae) productivity in Puerto Rico. AB - The hypothesis tested was that most pupae of Aedes aegypti are produced in a few types of containers so that vector control efforts could concentrate on eliminating the most productive ones and thus prevent dengue outbreaks. Pupal surveys were conducted twice in 2004 in an urban area in southern Puerto Rico. A total 35,030 immature mosquitoes (III and IV instars, pupae) was counted in 1,367 containers found with water in 624 premises during the first survey. Only pupae were counted in the second survey in 829 premises, 257 of which had containers with water, and 124 contained Ae. aegypti pupae (15%, 22% in the first survey). We found fewer (583) containers with water than in the first survey, but 202 had pupae (35%; 18.5% in first survey). Containers yielded 3,189 Ae. aegypti pupae, which was slightly fewer than those found in the first survey (3,388 pupae). The hypothesis was supported by the data, showing that 7 of 18 types of containers contained 80% of all female pupae. The most productive containers generally were also common. We used several criteria (i.e., container use, two-step cluster analysis based on environmental variables of containers and premises) to classify the containers and premises and to evaluate pupal distribution at various spatial scales (container, premise, and residences versus public areas). Most pupae were in 4 of 10 types of container usage categories. The cluster technique showed that most pupae were in unattended, rain-filled containers in the yards, particularly in receptacles in the shade of trees that received rainfall through foliage and had lower water temperatures. Pupal counts were adjusted to a negative binomial distribution, confirming their highly aggregated dispersal pattern. Cluster analysis showed that 61.3% of female pupae were in 40 (6.4%) of 624 premises that had in common their larger yards, number of trees, and container water volume. Using number of Ae. aegypti larvae, Breteau Index, or the presence of immature forms as indicators of pupal productivity is not as efficient in identifying the most productive types of containers as direct pupal counts. PMID- 16474087 TI - Identification in triatomine vectors of feeding sources and Trypanosoma cruzi variants by heteroduplex assay and a multiplex miniexon polymerase chain reaction. AB - Feeding sources of triatomine vectors (Triatoma longipennis) collected in peridomiciles in Mexico were identified by a heteroduplex assay developed with triatomine blood meals. Trypanosoma cruzi parasites were also characterized in the same blood meal samples by multiplex-polymerase chain reaction assay of mini exon gene inter-genic regions. The main blood meal source was from rats, but the bugs were able to feed on a wide variety of hosts, and human blood meals were identified. Trypanosoma cruzi was the only flagellate species identified in the blood meals. All populations belong to the T. cruzi I lineage, a result that is consistent with the previously assumed predominance of this lineage in Mexico. This combination of blood meal and T. cruzi lineage identification provides a powerful tool for understanding T. cruzi transmission cycles. PMID- 16474088 TI - Leishmania tropica-isolated patient with visceral leishmaniasis in southern Iran. AB - Visceral leishmaniasis (VL) is caused by various strains of Leishmania donovani, Leishmania infantum, and Leishmania chagasi with different geographical distribution. The aim of this study was to identify the strains of Leishmania that can cause VL in southern Iran. DNA of Leishmania were extracted from the slides of bone marrow aspirates (#42) and spleen punctures (#22), which were positive for leishman body from the patients who were referred to the hospitals affiliated with Shiraz University of Medical Sciences. Differences in Leishmania strains were determined by size difference of the polymerase chain reaction (PCR) amplification as visualized on agarose gel. PCR results and smears had 100% correlation. The dominant strain of Leishmania was L. infantum (63 out of the 64 cases), but one case of L. tropica was also detected. VL mostly involves children below 2 years of age in Iran, therefore infection with L. infantum was expected, but this study is the first report of VL that is caused by L. tropica in Iran. PMID- 16474089 TI - Spatial clustering and epidemiological aspects of visceral leishmaniasis in two endemic villages, Baringo District, Kenya. AB - Visceral leishmaniasis (VL) seroprevalence in Kenya is unknown because of the lack of a practical and accurate diagnostic test or surveillance system. A novel serological assay was used to estimate the seroprevalence of Leishmania-specific antibodies, and Global Information System and spatial clustering techniques were applied to study the presence of spatial clusters in Parkarin and Loboi villages in Baringo District in 2001. VL seroprevalences were 52.5% in Parkarin and 16.9% in Loboi. Significant associations among seropositivity and house construction, age, and proximity to domestic animal enclosures were found. A significant spatial cluster of VL was found in Loboi. The spatial distribution of cases in the two villages was different with respect to risk factors, such as presence of domestic animals. This study suggests that disease control efforts could be focused on elimination of sand fly habitat, placement of domestic animal enclosures, and targeted use of insecticides. PMID- 16474090 TI - Absence of leishmania in Guianan bats. AB - Studying the ecology of Leishmania parasites is essential for understanding and controlling the epidemiology of the diseases they cause. Despite their abundance and diversity in neotropical forests, few studies have been conducted to investigate the potential involvement of Chiroptera in the Leishmania pathogenic complexes. However, phlebotomine sand flies are known to colonize the same anthropized habitat, are attracted to bats, and are able to transmit trypanosomatids. Thus, 216 bats representing 29 species were sampled in the field in different primary and secondary forests of French Guiana where human cutaneous leishmaniases have been reported, together with 62 non-volant mammals. A series of 411 tissue samples representing 47 mammalian species were cultured and screened for the presence of Leishmania spp. by a genus-specific polymerase chain reaction. All 278 individuals surveyed were negative. Thus, bats do not appear to be involved in the Leishmania parasitic cycles in the Guyanas. PMID- 16474091 TI - Prevalence of cryptosporidium and other enteric parasites among wild non-human primates in Polonnaruwa, Sri Lanka. AB - Cryptosporidiosis is a rapidly emerging disease in the tropics. This is the first report of Cryptosporidium and other protozoan infections (Entamoeba spp., Iodamoeba, Chilomastix, and Balantidium spp.) in wild primates that inhabit the natural forest of Sri Lanka. It is unclear if non-human primates serve as a reservoir for these parasites under certain conditions. A cross-sectional coprologic survey among 125 monkeys (89 toque macaques, 21 gray langurs, and 15 purple-faced langurs) indicated that Cryptosporidium was detected in all three primate species and was most common among monkeys using areas and water that had been heavily soiled by human feces and livestock. Most macaques (96%) shedding Cryptosporidium oocysts were co-infected with other protozoans and important anthropozoonotic gastrointestinal parasites (e.g., Enterobius and Strongyloides). The transmission of these parasites among primates in the wild may have important implications for public health as well as wildlife conservation management. PMID- 16474092 TI - Indirect hemagglutination assay in patients with melioidosis in northern Australia. AB - Melioidosis is caused by the saprophytic organism Burkholderia pseudomallei. The use of the indirect hemagglutination assay (IHA) has found widespread use in areas endemic for this disease. Using this assay, we explored the serologic profile of 275 patients with culture-confirmed melioidosis in the Northern Territory of Australia. Based on a threshold titer of 1:40, the sensitivity of the IHA on admission was 56%. Female patients, those with positive blood cultures, and those with pneumonia independently predicted a negative IHA result. Most patients (68%) with negative admission IHA titers subsequently seroconverted. Most patients (92%) with positive admission IHA titers had persistently positive IHA titers. Relapses were not observed in 36 patients who had a negative IHA at least 1 month after admission, irrespective of initial admission IHA. The IHA has limited utility as a diagnostic test for acute disease, and most patients subsequently have persistently positive titers after recovery from illness. PMID- 16474093 TI - A drug-based intervention study on the importance of buffaloes for human Schistosoma japonicum infection around Poyang Lake, People's Republic of China. AB - Schistosomiasis japonica is a zoonosis of major public health importance in southern China. We undertook a drug intervention to test the hypothesis that buffalo are major reservoirs for human infection in the marshlands/lake areas, where one million people are infected. We compared human and buffalo infection rates and intensity in an intervention village (Jishan), where humans and buffalo were treated with praziquantel, and a control village (Hexi), where only humans were treated, in the Poyang Lake region. Over the four-year study, human incidence in Jishan decreased but increased in Hexi. Adjustment of incidence by age, sex, water exposure, year, and village further confirmed the decreased human infection in Jishan. Chemotherapy for buffaloes resulted in a decrease in buffalo infection rates in Jishan, which coincided with the reduction in human infection rates there in the last two years of the study. Mathematical modeling predicted that buffalo are responsible for 75% of human transmission in Jishan. PMID- 16474094 TI - Failure of standard treatment with praziquantel in two returned travelers with Schistosoma haematobium infection. AB - A single 40 mg/kg dose of praziquantel (PZQ) continues to be the standard treatment for schistosomiasis caused by S. mansoni and S. haematobium in all clinical settings. Experimental development of drug resistance and the recent isolation of S. mansoni strains with a natural tolerance to high doses of PZQ have raised concerns over the adequacy of such a dose. We describe two Spanish travelers with genitourinary schistosomiasis caused by S. haematobium in whom repeated standard treatment failed to clear the infection. PMID- 16474095 TI - Bactericidal function of alveolar macrophages in mechanically ventilated rabbits. AB - Protective ventilation strategies have been universally embraced because of reduced mortality. We tested the hypothesis that tidal volume (VT) in an in vivo model of mechanical ventilation would modulate bactericidal function of alveolar macrophages (AMs). Adult New Zealand White rabbits were mechanically ventilated for 4 h with a VT of 6 ml/kg (low) or a VT of 12 ml/kg (traditional), with each group receiving 3 cm H2O positive end-expiratory pressure with and without intratracheal lipopolysaccharide (LPS) instillation (20 mg/kg). AMs were isolated from bronchoalveolar lavage fluid taken from the whole left lung and used for bacterial killing assays. There were no significant differences in steady-state levels of nitrite or AM phagocytosis and killing of Klebsiella pneumoniae, although these values trended to be slightly higher in the traditional VT group. However, bronchoalveolar lavage fluid protein concentrations were significantly increased in traditional VT groups receiving LPS compared with animals ventilated with a low VT (1,407.8 +/- 121.4 versus 934.7 +/- 118.2; P < 0.001). Lung wet:dry weight ratio in the traditional VT group was increased when compared with the low VT group without LPS (7.3 +/- 0.4 versus 6.1 +/- 0.3, respectively; P < 0.05). Additionally, IL-8 expression was significantly greater under conditions of LPS treatment and mechanical ventilation at VT of 12 ml/kg. These results suggest that the traditional ventilator approach (12 ml/kg VT) in a model of in vivo mechanical ventilation results in lung pathology without affecting AM antibacterial function. PMID- 16474097 TI - Multistrain genetic comparisons reveal CCR5 as a receptor involved in airway hyperresponsiveness. AB - Asthma is a ubiquitous disease with a broad range of clinical phenotypes. To better understand the complex genetic and environmental interactions underlying asthma, we compared the gene-gene interactions of four genetically distinct mouse strains that demonstrate biologically distinct responses to allergen. Using DNA microarrays and knock-out mouse studies, we showed that CCR5 plays a definitive role in the development of ovalbumin-induced allergic airway inflammatory disease. In addition, gene expression profiling data have revealed other potential novel targets for therapeutics-based research and has enhanced the understanding of the molecular mechanisms underlying the etiology of "asthma." PMID- 16474096 TI - Tissue-specific renin-angiotensin system in pulmonary lymphangioleiomyomatosis. AB - Lymphangioleiomyomatosis (LAM), a multisystem disease found in middle-aged women, is characterized by cystic lung destruction and abdominal tumors (e.g., angiomyolipomas, lymphangioleimyomas), resulting from proliferation of abnormal appearing, smooth muscle-like cells (LAM cells). The LAM cells, in combination with other cells, form nodular structures within the lung interstitium and in the walls of the cysts. LAM cells contain mutations in the tuberous sclerosis complex TSC1 and/or TSC2 genes, which lead to dysregulation of the mammalian target of rapamycin, affecting cell growth and proliferation. Proliferation and migration of vascular smooth muscle cells and production of angiogenic factors are regulated, in part, by angiotensin II. To determine whether a LAM-specific renin angiotensin system might play a role in the pathogenesis of LAM, we investigated the expression of genes and gene products of this system in LAM nodules. mRNA for angiotensinogen was present in RNA isolated by laser-captured microdissection from LAM nodules. Angiotensin I-converting enzyme and chymase-producing mast cells were present within the LAM nodules. We detected renin in LAM cells, as determined by the presence of mRNA and immunohistochemistry. Angiotensin II type 1 and type II receptors were identified in LAM cells by immunohistochemistry and immunoblotting of microdissected LAM nodules. Angiotensin II is localized in cells containing alpha-smooth muscle actin (LAM cells). A LAM-specific renin angiotensin system appears to function within the LAM nodule as an autocrine system that could promote LAM cell proliferation and migration, and could represent a pharmacologic target. PMID- 16474098 TI - Role of granulocyte macrophage colony-stimulating factor during gram-negative lung infection with Pseudomonas aeruginosa. AB - Granulocyte macrophage colony-stimulating factor (GM-CSF) stimulates survival, proliferation, differentiation, and function of myeloid cells. Recently, GM-CSF has been shown to be important for normal pulmonary homeostasis. We report that GM-CSF is induced in lung leukocytes during infection with Gram-negative bacteria. Therefore, we postulated that deficiencies in GM-CSF would increase susceptibility to Gram-negative infection in vivo. After an intratracheal inoculum with Pseudomonas aeruginosa, GM-CSF-/- mice show decreased survival compared with wild-type mice. GM-CSF-/- mice show increased lung, spleen, and blood bacterial CFU. GM-CSF-/- mice are defective in the production of cysteinyl leukotrienes, prostaglandin E2, macrophage inflammatory protein, and keratinocyte derived chemokine in lung leukocytes postinfection. Despite these defects, inflammatory cell recruitment is not diminished at 6 or 24 h postinfection, and the functional activity of polymorphonuclear leukocytes from the lung and peritoneum against P. aeruginosa is enhanced in GM-CSF-/- mice. In contrast, alveolar macrophage (AM) phagocytosis, killing, and H2O2 production are defective in GM-CSF-/- mice. Although the absence of GM-CSF has profound effects on AMs, peritoneal macrophages seem to have normal bactericidal activities in GM-CSF-/- mice. Defects in AM function may be related to diminished levels of IFN-gamma and TNF-alpha postinfection. Thus, GM-CSF-/- mice are more susceptible to lung infection with P. aeruginosa as a result of impaired AM function. PMID- 16474099 TI - Gene induction during differentiation of human pulmonary type II cells in vitro. AB - Mature alveolar type II cells that produce pulmonary surfactant are essential for adaptation to extrauterine life. We profiled gene expression in human fetal lung epithelial cells cultured in serum-free medium containing dexamethasone and cyclic AMP, a treatment that induces differentiation of type II cells. Microarray analysis identified 388 genes that were induced > 1.5-fold by 72 h of hormone treatment. Induced genes represented all categories of molecular function and subcellular location, with increased frequency in the categories of ionic channel, cell adhesion, surface film, lysosome, extracellular matrix, and basement membrane. In time-course experiments, self-organizing map analysis identified a cluster of 17 genes that were slowly but highly induced (5- to approximately 190-fold) and represented four functional categories: surfactant related (SFTPC, SFTPA, PGC, SFTPB, LAMP3, LPL), regulatory (WIF2, IGF2, IL1RL1, NR4A2, HIF3A), metabolic (MAOA, ADH1B, SEPP1), and transport (SCNN1A, CLDN18, AQP4). Induction of both mRNA and protein for these genes, which included nine newly identified regulated genes, was confirmed, and cellular localization was determined in both fetal and postnatal tissue. Induction of lysosomal-associated membrane protein 3 required both hormones, and expression was localized to limiting membranes of lamellar bodies. Hormone-induced differentiation of human type II cells is associated with genome-wide increased expression of genes with diverse functions. PMID- 16474101 TI - Novel mechanism for impaired nitrergic relaxation in achalasia. PMID- 16474102 TI - To feed or not to feed? Are nutritional supplements worthwhile in active Crohn's disease? PMID- 16474100 TI - Use and abuse of hydrogen breath tests. AB - Hydrogen breath tests are widely used to explore the pathophysiology of functional gastrointestinal disorders. Small intestinal bacterial overgrowth and carbohydrate malabsorption are disorders detected by these tests that have been proposed to be of great importance for symptoms in, for instance, irritable bowel syndrome. However, conclusions drawn from these studies are highly controversial and divergent results exist. There is also an extensive use of these tests in clinical practice with difficulties regarding interpretation of the tests and sometimes erroneous conclusions. The limitations and pitfalls of these tests will be reviewed in this article, and hopefully the occasional abuse of these tests can be turned into proper clinical and scientific use instead in the future. PMID- 16474103 TI - Slipping the barrier: how variants in CARD15 could alter permeability of the intestinal wall and population health. PMID- 16474104 TI - Non-invasive evaluation of hepatic fibrosis: don't count your chickens before they're hatched. PMID- 16474105 TI - Inflammatory bowel disease--be aware of the mimics. Diagnosis: lymphogranuloma venerum. PMID- 16474106 TI - An unusual cause of gastric outlet obstruction. Diagnosis: Bourevet's syndrome. PMID- 16474107 TI - Chewing is an important first step in digestion. Diagnosis: piece of glass in stomach. PMID- 16474108 TI - Arterial hypertension in cirrhosis: arterial compliance, volume distribution, and central haemodynamics. AB - BACKGROUND AND AIMS: Arterial hypertension is a common disorder. Hyperkinetic circulation and reduced effective volaemia are central elements in the haemodynamic dysfunction in cirrhosis. The aim of the present study was to investigate whether cirrhotic patients with arterial hypertension are normokinetic and normovolaemic or whether they reveal the same circulatory dysfunction as their normotensive counterparts. MATERIAL AND METHODS: Thirty three patients with arterial hypertension were identified among 648 patients with cirrhosis: 14 in Child class A, 12 in class B, and seven in class C. Controls were 130 normotensive cirrhotic patients, 19 controls with normal arterial blood pressure and without liver disease, and 16 patients with essential arterial hypertension. All groups underwent haemodynamic investigation with determination of cardiac output (CO), plasma volume (PV), central blood volume (CBV), hepatic venous pressure gradient (HVPG), hepatic blood flow (HBF), arterial compliance (AC), and systemic vascular resistance (SVR) in the supine position. RESULTS: Liver function, as evaluated by galactose elimination capacity, indocyanine green clearance, HBF, and Child score, was significantly better in hypertensive cirrhotics than in their normotensive counterparts (p<0.05-0.01) but portal pressure was similar (HVPG 13 v 15 mm Hg; NS). AC was significantly lower and normal in the arterial hypertensive cirrhotic group (1.07 v 1.39 mm Hg/ml; p<0.02) and SVR was significantly higher and normal (1475 v 1020 dynxs/cm5; p<0.01). Arterial hypertensive cirrhotic patients were hyperdynamic (CO 6.80 v 7.14 l/min; NS) and central hypovolaemic (CBV 19.8 v 20.6 ml/kg; NS), as were normotensive patients, but differences were found in relation to arterial blood pressure. Whereas arterial pressure was inversely correlated with CO, PV, and Child score in the normotensive group (p< 0.01), the same correlations were either direct or insignificant in arterial hypertensive cirrhotics. CONCLUSION: Arterial hypertensive cirrhotic patients are hyperkinetic and central hypovolaemic, in common with their normotensive counterparts, but vasodilatation is reduced and regulation of arterial blood pressure may be less deranged. PMID- 16474109 TI - Laboratory markers in IBD: useful, magic, or unnecessary toys? AB - Laboratory markers have been investigated in inflammatory bowel disease (IBD) for diagnostic and differential diagnostic purposes, for assessment of disease activity and risk of complications, for prediction of relapse, and for monitoring the effect of therapy. The introduction of biological therapies in IBD has renewed interest in inflammatory markers (especially C reactive protein (CRP)), given their potential to select responders to these treatments. Of all the laboratory markers, CRP is the most studied and has been shown to have the best overall performance. CRP is an objective marker of inflammation and correlates well with disease activity in Crohn's disease (CD). Increased CRP levels are associated with better response rates and normal CRP levels predict high placebo response rates in clinical trials with biologicals. However, despite the advantages of CRP over other markers, it is still far from ideal. Furthermore, CRP correlates less well with disease activity in patients with ulcerative colitis (UC) as compared with CD. Other laboratory markers, including erythrocyte sedimentation rate (ESR), leucocyte and platelet count, albumin, and 1 acid glycoprotein (orosomucoid), have been studied either less extensively in IBD or have proven to be less useful than CRP. Faecal markers seem promising and may be more specific in detecting gut inflammation in patients with established IBD. Promising results have been reported with the use of faecal calprotectin in CD as well as in UC. Recent data however suggest that the performance of the faecal calprotectin test is superior for UC than for CD. Taken together, laboratory markers are useful and should be part of the global management of our IBD patients. They are however not magic and until more data become available, the use of CRP and other laboratory markers should be seen as an additive tool to clinical observation and physical examination rather than a replacement. PMID- 16474110 TI - Circulating ghrelin levels in patients with inflammatory bowel disease. PMID- 16474111 TI - Postprandial response of salivary ghrelin and leptin to carbohydrate uptake. PMID- 16474112 TI - Bravo wireless versus catheter pH monitoring systems. PMID- 16474113 TI - Complications of radiofrequency thermal ablation in hepatocellular carcinoma: what about "explosive" spread? PMID- 16474114 TI - Collagenous colitis in adult Sri Lankans: experience from the Indian subcontinent. PMID- 16474115 TI - Comparison of virus transcription during lytic infection of the Oka parental and vaccine strains of Varicella-Zoster virus. AB - The attenuated Oka vaccine (V-Oka) strain of varicella-zoster virus (VZV) effectively reduces disease produced by primary infection and virus reactivation. V-Oka was developed by propagation of the Oka parental (P-Oka) strain of VZV in guinea pig and human embryo fibroblasts. Complete DNA sequencing of both viruses has revealed 63 sites that differ between P-Oka and V-Oka, 37 of which are located within 21 unique open reading frames (ORFs). Of the ORFs that differ, ORF 62 contains the greatest number (10) of mutated sites. ORF 62 encodes IE 62, the major immediate-early transactivator of virus genes, and is essential for lytic virus replication. To determine whether a disproportionate number of mutations in ORF 62 might account for virus attenuation, we compared the global pattern of V Oka gene expression to that of P-Oka. Transcription of ORFs 62, 65, 66, and 67 was suppressed, whereas ORF 41 was elevated in V-Oka-infected cells compared to P Oka-infected cells (P < 0.01; z test). Suppression of ORF 62, 65, and 66 transcription was confirmed by quantitative dot blot and Western blot analyses. Transient-transfection assays to determine whether mutations within V-Oka-derived IE 62 affected its ability to transactivate VZV gene promoters revealed similar IE 62 transactivation of VZV gene 20, 21, 28, 29, 65, and 66 promoters in both P Oka and V-Oka. Together, our results indicate that mutations in V-Oka IE 62 alone are unlikely to account for vaccine virus attenuation. PMID- 16474117 TI - Human immunodeficiency virus (HIV) vaccine trials: a novel assay for differential diagnosis of HIV infections in the face of vaccine-generated antibodies. AB - All current human immunodeficiency virus (HIV) vaccine candidates contain multiple viral components and elicit antibodies that react positively in licensed HIV diagnostic tests, which contain similar viral products. Thus, vaccine trial participants could be falsely diagnosed as infected with HIV. Additionally, uninfected, seropositive vaccinees may encounter long-term social and economic harms. Moreover, this also interferes with early detection of true HIV infections during preventive HIV vaccine trials. An HIV-seropositive test result among uninfected vaccine trial participants is a major public health concern for volunteers who want to participate in future HIV vaccine trials. Based on the increased number of HIV vaccines being tested globally, it is essential to differentiate vaccine- from virus-induced antibodies. Using a whole-HIV-genome phage display library, we identified conserved sequences in Env-gp41 and Gag-p6 which are recognized soon after infection, do not contain protective epitopes, and are not part of most current HIV vaccines. We established a new HIV serodetection assay based on these peptides. To date, this assay, termed HIV SELECTEST, demonstrates >99% specificity and sensitivity. Importantly, in testing of plasma samples from multiple HIV vaccine trials, uninfected trial participants scored negative, while all intercurrent infections were detected within 1 to 3 months of HIV infection. The new HIV-SELECTEST is a simple but robust diagnostic tool for easy implementation in HIV vaccine trials and blood banks worldwide. PMID- 16474116 TI - Stability and function of JC virus large T antigen and T' proteins are altered by mutation of their phosphorylated threonine 125 residues. AB - JC virus (JCV), a human polyomavirus, exhibits oncogenic activity in rodents and primates. The large tumor antigens (TAgs) of the polyomaviruses play key roles in viral replication and oncogenic transformation. Analyses of JCV TAg phosphorylation mutants indicated that the amino-terminal phosphorylation site at threonine 125 (T125) is critical to TAg replication function. This site is also conserved in the TAg splice variants T'(135), T'(136), and T'(165). By constructing stable cell lines expressing JCV T125A and T125D mutants, we show that mutation of this phosphorylation site to alanine generates an unstable TAg; however, the stability of the three T' proteins is unaffected. JCV T125A mutant proteins bind the retinoblastoma protein (RB) family members p107 and p130 with slightly reduced efficiencies and fail to induce the release of transcriptionally active E2F from RB-E2F complexes. On the other hand, cell lines expressing JCV T125D mutant proteins produce stable TAg and T' proteins which bind p107 and p130 more efficiently than do the wild-type proteins. In addition, T125D mutant proteins efficiently induce the release of E2F from RB-E2F complexes. T125D mutant cell lines, unlike the T125A mutant lines, continue to grow under conditions of low serum concentration and anchorage independence. Finally, both T125A and T125D mutant viruses are replication defective. Phosphorylation of the T125 site is likely mediated by a cyclin-cyclin-dependent kinase, suggesting that JCV TAg and T' protein functions that mediate viral replication and oncogenic transformation events are regulated in a cell cycle-dependent manner. PMID- 16474118 TI - Retroviral restriction factors Fv1 and TRIM5alpha act independently and can compete for incoming virus before reverse transcription. AB - The restriction factors Fv1 and TRIM5alpha provide dominant blocks to retroviral infection, targeting incoming capsids at a postentry, preintegration step. They both restrict N-tropic murine leukemia virus with similar specificity yet act at different points in the viral life cycle. TRIM5alpha-restricted virus is usually unable to reverse transcribe, whereas Fv1-restricted virus reverse transcribes normally. Here we investigate the relationship between these two restriction factors by expressing Fv1 alleles in human cells. We demonstrate that Fv1 is able to compete with TRIM5alpha for virus before reverse transcription. In human cells expressing Fv1(b), N-tropic restricted virus becomes less infectious but reverse transcribes more efficiently, indicating competition between the two antiviral molecules and protection of the virus from TRIM5alpha by Fv1. Our findings suggest that, like TRIM5alpha, Fv1 interacts with virus before reverse transcription, but the consequences of this interaction are not realized until a later stage of the life cycle. We also demonstrate that Fv1 is functionally independent of TRIM5alpha when expressed in human cells. PMID- 16474119 TI - Mouse-adapted scrapie infection of SN56 cells: greater efficiency with microsome associated versus purified PrP-res. AB - The process by which transmissible spongiform encephalopathy agents, or prions, infect cells is unknown. We employed a new differentiable cell line (SN56) susceptible to infection with three mouse-adapted scrapie strains to gain insight into the cellular infection process. The effect of disease-associated PrP (PrP res) association with microsomal membranes on infection efficiency was examined by comparing sustained PrP-res production in cells treated with either scrapie brain microsomes or purified, detergent-extracted PrP-res. When normalized for quantity of input PrP-res, scrapie brain microsomes induced dramatically enhanced persistent PrP-res formation compared to purified PrP-res. Infected SN56 cells released low levels of PrP-res into the culture supernatant, which also efficiently initiated infection in recipient cells. Interestingly, microsomes labeled with a fluorescent marker were internalized by SN56 cells in small vesicles, which were subsequently found in neuritic processes. When bound to culture wells to reduce internalization during the infection process, scrapie microsomes induced less long-term PrP-res production than suspended microsomes. Long-term differentiation of infected SN56 cells was accompanied by a decrease in PrP-res formation. Our observations provide evidence that infection of cells is aided by the association of PrP-res with membranes and/or other microsomal constituents. PMID- 16474120 TI - Herpes simplex virus type 1 DNA-packaging protein UL17 is required for efficient binding of UL25 to capsids. AB - Herpes simplex virus type 1 packages its DNA genome into a precursor capsid, referred to as the procapsid. Of the three capsid-associated DNA-packaging proteins, UL17, UL25, and UL6, only UL17 and UL6 appear to be components of the procapsid, with UL25 being added subsequently. To determine whether the association of UL17 or UL25 with capsids was dependent on the other two packaging proteins, B capsids, which lack viral DNA but retain the cleaved internal scaffold, were purified from nonpermissive cells infected with UL17, UL25, or UL6 null mutants and compared with wild-type (wt) B capsids. In the absence of UL17, the levels of UL25 in the mutant capsids were much lower than those in wt B capsids. These results suggest that UL17 is required for efficient incorporation of UL25 into B capsids. B capsids lacking UL25 contained about twofold-less UL17 than wt capsids, raising the possibilities that UL25 is important for stabilizing UL17 in capsids and that the two proteins interact in the capsid. The distribution of UL17 and UL25 on B capsids was examined using immunogold labeling. Both proteins appeared to bind to multiple sites on the capsid. The properties of the UL17 and UL25 proteins are consistent with the idea that the two proteins are important in stabilizing capsid-DNA structures rather than having a direct role in DNA packaging. PMID- 16474122 TI - Hepatitis B virus reverse transcriptase and epsilon RNA sequences required for specific interaction in vitro. AB - Initiation of reverse transcription and nucleocapsid assembly in hepatitis B virus (HBV) depends on the specific recognition of an RNA signal (the packaging signal, epsilon) on the pregenomic RNA by the viral reverse transcriptase (RT). Using an in vitro reconstitution system whereby the cellular heat shock protein 90 chaperone system activates recombinant HBV RT for specific epsilon binding, we have defined the protein and RNA sequences required for specific HBV RT-epsilon interaction in vitro. Our results indicated that approximately 150 amino acid residues from the terminal protein domain and 230 from the RT domain were necessary and sufficient for epsilon binding. With respect to the epsilon RNA sequence, its internal bulge and, in particular, the first nucleotide (C) of the bulge were specifically required for RT binding. Sequences from the upper portion of the lower stem and the lower portion of the upper stem also contributed to RT binding, as did the base pairing of the upper portion and the single unpaired U residue of the upper stem. Surprisingly, the apical loop of epsilon, known to be required for RNA packaging, was entirely dispensable for RT binding. A comparison of the requirements for in vitro RT-epsilon interaction with those for in vivo pregenomic RNA (pgRNA) packaging clearly indicated that RT-epsilon interaction was necessary but not sufficient for pgRNA packaging. In addition, our results suggest that recognition of some epsilon sequences by the RT may be required specifically for viral DNA synthesis. PMID- 16474123 TI - Comparison of the replication of influenza A viruses in Chinese ring-necked pheasants and chukar partridges. AB - We investigated the replication and transmission of avian influenza A viruses in two species thought to be intermediate hosts in the spread of influenza A viruses in live poultry markets: Chinese ring-necked pheasants and chukar partridges. All 15 hemagglutinin subtypes replicated in pheasants, and most subtypes transmitted to naive contact pheasants, primarily via the fecal-oral route. Many viruses were shed from the gastrointestinal tract of experimentally inoculated pheasants for 14 days or longer. Virus was isolated from the cloacal swabs of one contact pheasant for an unprecedented 45 days. Chukar partridges were less susceptible to infection with avian influenza viruses. The viruses that replicated in chukar partridges were isolated for 7 days after experimental inoculation, predominantly from the respiratory tract. We detected high neutralizing antibody titers with correspondingly low levels of serum hemagglutination inhibition antibody titers in pheasants and chukar partridges when chicken red blood cells were used in serological analyses. When horse erythrocytes were used, antibody titers were comparable to those obtained by using the neutralization assay. More importantly, the results suggested that pheasants can serve as a reservoir of influenza virus. Because of their continuous asymptomatic infection and longer stay in the markets, pheasants are ideal "carriers" of influenza A viruses. Their continued presence in live markets contributes to the perpetuation and genetic interaction of influenza viruses there. On the basis of our findings, it does not make good sense to ban quail but not pheasants from the live markets. PMID- 16474121 TI - Vaccinia virus proteome: identification of proteins in vaccinia virus intracellular mature virion particles. AB - Vaccinia virus is a large enveloped poxvirus with more than 200 genes in its genome. Although many poxvirus genomes have been sequenced, knowledge of the host and viral protein components of the virions remains incomplete. In this study, we used gel-free liquid chromatography and tandem mass spectroscopy to identify the viral and host proteins in purified vaccinia intracellular mature virions (IMV). Analysis of the proteins in the IMV showed that it contains 75 viral proteins, including structural proteins, enzymes, transcription factors, and predicted viral proteins not known to be expressed or present in the IMV. We also determined the relative abundances of the individual protein components in the IMV. Finally, 23 IMV-associated host proteins were also identified. This study provides the first comprehensive structural analysis of the infectious vaccinia virus IMV. PMID- 16474124 TI - Proteomics analysis of the tombusvirus replicase: Hsp70 molecular chaperone is associated with the replicase and enhances viral RNA replication. AB - Plus-strand RNA virus replication occurs via the assembly of viral replicase complexes involving multiple viral and host proteins. To identify host proteins present in the cucumber necrosis tombusvirus (CNV) replicase, we affinity purified functional viral replicase complexes from yeast. Mass spectrometry analysis of proteins resolved by two-dimensional gel electrophoresis revealed the presence of CNV p33 and p92 replicase proteins as well as four major host proteins in the CNV replicase. The host proteins included the Ssa1/2p molecular chaperones (yeast homologues of Hsp70 proteins), Tdh2/3p (glyceraldehyde-3 phosphate dehydrogenase, an RNA-binding protein), Pdc1p (pyruvate decarboxylase), and an unknown approximately 35-kDa acidic protein. Copurification experiments demonstrated that Ssa1p bound to p33 replication protein in vivo, and surface plasmon resonance measurements with purified recombinant proteins confirmed this interaction in vitro. The double mutant strain (ssa1 ssa2) showed 75% reduction in viral RNA accumulation, whereas overexpression of either Ssa1p or Ssa2p stimulated viral RNA replication by approximately threefold. The activity of the purified CNV replicase correlated with viral RNA replication in the above mentioned ssa1 ssa2 mutant and in the Ssa overexpression strains, suggesting that Ssa1/2p likely plays an important role in the assembly of the CNV replicase. PMID- 16474125 TI - RNA secondary structure in the coding region of dengue virus type 2 directs translation start codon selection and is required for viral replication. AB - Dengue virus is a positive-strand RNA virus and a member of the genus Flavivirus, which includes West Nile, yellow fever, and tick-borne encephalitis viruses. Flavivirus genomes are translated as a single polyprotein that is subsequently cleaved into 10 proteins, the first of which is the viral capsid (C) protein. Dengue virus type 2 (DENV2) and other mosquito-borne flaviviruses initiate translation of C from a start codon in a suboptimal context and have multiple in frame AUGs downstream. Here, we show that an RNA hairpin structure in the capsid coding region (cHP) directs translation start site selection in human and mosquito cells. The ability of the cHP to direct initiation from the first start codon is proportional to its thermodynamic stability, is position dependent, and is sequence independent, consistent with a mechanism in which the scanning initiation complex stalls momentarily over the first AUG as it begins to unwind the cHP. The cHP of tick-borne flaviviruses is not maintained in a position to influence start codon selection, which suggests that this coding region cis element may serve another function in the flavivirus life cycle. Here, we demonstrate that the DENV2 cHP and both the first and second AUGs of C are necessary for efficient viral replication in human and mosquito cells. While numerous regulatory elements have been identified in the untranslated regions of RNA viral genomes, we show that the cHP is a coding-region RNA element that directs start codon selection and is required for viral replication. PMID- 16474127 TI - Matrix protein mutant of vesicular stomatitis virus stimulates maturation of myeloid dendritic cells. AB - Matrix (M) protein mutants of vesicular stomatitis virus have recently been used as oncolytic viruses for tumor therapies and are being developed as vaccine vectors for heterologous antigens. Because dendritic cell (DC) maturation is an important correlate of tumor immunosurveillance and vaccine efficacy, we sought to determine the ability of a recombinant M protein mutant virus (rM51R-M virus) to mature DC in vitro. We have previously shown that rM51R-M virus is defective at inhibiting host gene expression in several cell lines compared to its recombinant wild-type counterpart, rwt virus. Therefore, rM51R-M virus allows the expression of genes involved in antiviral responses, such as the type I interferon (IFN) gene. Our results demonstrate that, in contrast to the rwt virus, rM51R-M virus induced the maturation of myeloid DC (mDC) populations, as indicated by an increase in the surface expression of CD40, CD80, and CD86 as well as the secretion of interleukin-12 (IL-12), IL-6, and type I IFN. In addition, mDC infected with rM51R-M virus effectively activated naive T cells in vitro, whereas rwt virus-infected mDC were defective in antigen presentation. The inability of rwt virus to induce mDC maturation was correlated with the inhibition of host gene expression in rwt virus-infected cells. Our studies also indicated that the production of costimulatory molecules on mDC by rM51R-M virus was dependent on the type I IFN receptor, while maturation induced by this virus was largely independent of MyD88. These data indicate that rM51R-M virus effectively stimulates the maturation of mDC and has the potential to promote effective T-cell responses to vector-expressed antigens, activate DC at tumor sites during therapy, and aid in tumor immunosurveillance and destruction. PMID- 16474126 TI - Bovine coronavirus 5'-proximal genomic acceptor hotspot for discontinuous transcription is 65 nucleotides wide. AB - Coronaviruses are positive-strand, RNA-dependent RNA polymerase-utilizing viruses that require a polymerase template switch, characterized as discontinuous transcription, to place a 5'-terminal genomic leader onto subgenomic mRNAs (sgmRNAs). The usually precise switch is thought to occur during the synthesis of negative-strand templates for sgmRNA production and to be directed by heptameric core donor sequences within the genome that match an acceptor core (UCUAAAC in the case of bovine coronavirus) near the 3' end of the 5'-terminal genomic leader. Here it is shown that a 22-nucleotide (nt) donor sequence engineered into a packageable bovine coronavirus defective interfering (DI) RNA and made to match a sequence within the 65-nt virus genomic leader caused a template switch yielding an sgmRNA with only a 33-nt minileader. By changing the donor sequence, acceptor sites between genomic nt 33 and 97 (identical between the DI RNA and the viral genome) could be used to generate sgmRNAs detectable by Northern analysis (approximately 2 to 32 molecules per cell) by 24 h postinfection. Whether the switch was intramolecular only was not determined since a potentially distinguishing acceptor region in the DI RNA rapidly conformed to that in the helper virus genome through a previously described template switch known as leader switching. These results show that crossover acceptor sites for discontinuous transcription (i) need not include the UCUAAAC core and (ii) rest within a surprisingly wide 5'-proximal "hotspot." Overlap of this hotspot with that for leader switching and with elements required for RNA replication suggests that it is part of a larger 5'-proximal multifunctional structure. PMID- 16474128 TI - Gradual elimination of retroviruses in YBR/Ei mice. AB - Mouse mammary tumor virus (MMTV), a well-characterized retrovirus that causes mammary tumors in susceptible mice, is commonly used to investigate virus-host interactions. We have shown that YBR/Ei mice demonstrate a novel, dominant mechanism of resistance to MMTV infection and MMTV-induced mammary tumors. MMTV can both establish infection in YBR/Ei mice and be transmitted by YBR/Ei mice as an infectious virus. However, virus production is severely attenuated, resulting in gradual clearance of infection in successive generations. Our transfer experiments showed that T cells generated in MMTV-infected resistant mice were required to restrict MMTV replication in susceptible mice. These results emphasize the importance of inducing T-cell responses for effective protection against retroviral infections. PMID- 16474129 TI - Heptad repeat 2 in herpes simplex virus 1 gH interacts with heptad repeat 1 and is critical for virus entry and fusion. AB - Herpes simplex virus 1 (HSV-1) entry into cells and cell-cell fusion mediated by HSV-1 glycoproteins require four glycoproteins, gD, gB, gH, gL. Of these, gH is the only one that so far exhibits structural-functional features typical of viral fusion glycoproteins, i.e., a candidate fusion peptide and, downstream of it, a heptad repeat (HR) segment able to form a coiled coil, named HR-1. Here, we show that gH carries a functional HR-2 capable of physical interaction with HR-1. Specifically, mutational analysis of gH aimed at increasing or decreasing the ability of HR-2 to form a coiled coil resulted in an increase or decrease of fusion activity, respectively. HSV infection was modified accordingly. A mimetic peptide with the HR-2 sequence inhibited HSV-1 infection in a specific and dose dependent manner. Circular dichroism spectroscopy showed that both HR-2 and HR-1 mimetic peptides adopt mainly random conformation in aqueous solution, while a decrease in peptide environmental polarity determines a conformational change, with a significant increase of the alpha-helical conformation content, in particular, for the HR-1 peptide. Furthermore, HR-1 and HR-2 mimetic peptides formed a stable complex, as revealed in nondenaturing electrophoresis and by circular dichroism. The mixture of HR-1 and HR-2 peptides reversed the inhibition of HSV infection exerted by the single peptides. Complex formation between HR-1 and HR-2 was independent of the presence of adjacent gH sequences and of additional glycoproteins involved in entry and fusion. Altogether, HR-2 adds to the features typical of class 1 fusion glycoproteins exhibited by HSV-1 gH. PMID- 16474130 TI - Recognition of vaccinia virus-infected cells by human natural killer cells depends on natural cytotoxicity receptors. AB - Natural Killer (NK) cells are important in the immune response to a number of viruses; however, the mechanisms used by NK cells to discriminate between healthy and virus-infected cells are only beginning to be understood. Infection with vaccinia virus provokes a marked increase in the susceptibility of target cells to lysis by NK cells, and we show that recognition of the changes in the target cell induced by vaccinia virus infection depends on the natural cytotoxicity receptors NKp30, NKp44, and NKp46. Vaccinia virus infection does not induce expression of ligands for the activating NKG2D receptor, nor does downregulation of major histocompatibility complex class I molecules appear to be of critical importance for altered target cell susceptibility to NK cell lysis. The increased susceptibility to lysis by NK cells triggered upon poxvirus infection depends on a viral gene, or genes, transcribed early in the viral life cycle and present in multiple distinct orthopoxviruses. The more general implications of these data for the processes of innate immune recognition are discussed. PMID- 16474131 TI - Transcriptional origin of Kaposi's sarcoma-associated herpesvirus microRNAs. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) encodes 11 distinct microRNAs, all of which are found clustered within the major latency-associated region of the KSHV genome in the same transcriptional orientation. Because the KSHV microRNAs are all expressed in latently infected cells and are largely unaffected by induction of lytic replication, it appeared probable that they would be processed out of KSHV transcripts that are derived from a latent promoter(s) present in this region. Here, we define three latent transcripts, derived from two distinct KSHV latent promoters, that function as both KSHV primary microRNA precursors and as kaposin pre-mRNAs. These activities require the readthrough of a leaky viral polyadenylation signal located at nucleotide 122070 in the KSHV genome. In contrast, recognition of this polyadenylation signal gives rise to previously identified mRNAs that encode the KSHV open reading frames (ORFs) 71, 72 and 73 proteins as well as a novel unspliced KSHV mRNA that encodes only ORF72 and ORF71. Thus, transcripts initiating at the two latent promoters present in the KSHV latency-associated region can undergo two entirely distinct fates, i.e., processing to give a kaposin mRNA and viral microRNAs on the one hand or expression as KSHV ORF71, ORF72, or ORF73 mRNAs on the other, depending on whether the viral polyadenylation site located at position 122070 is ignored or recognized, respectively. PMID- 16474132 TI - Latency-associated nuclear antigen (LANA) of Kaposi's sarcoma-associated herpesvirus interacts with origin recognition complexes at the LANA binding sequence within the terminal repeats. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) DNA persists in latently infected cells as an episome via tethering to the host chromosomes. The latency-associated nuclear antigen (LANA) of KSHV binds to the cis-acting elements in the terminal repeat (TR) region of the genome through its carboxy terminus. Previous studies have demonstrated that LANA is important for episome maintenance and replication of the TR-containing plasmids. Here we report that LANA associates with origin recognition complexes (ORCs) when bound to its 17-bp LANA binding cognate sequence (LBS). Chromatin immunoprecipitation of multiple regions across the entire genome from two KSHV-infected cell lines, BC-3 and BCBL-1, revealed that the ORCs predominantly associated with the chromatin structure at the TR as well as two regions within the long unique region of the genome. Coimmunoprecipitation of ORCs with LANA-specific antibodies shows that ORCs can bind and form complexes with LANA in cells. This association was further supported by in vitro binding studies which showed that ORCs associate with LANA predominantly through the carboxy-terminal DNA binding region. KSHV-positive BC-3 and BCBL-1 cells arrested in G(1)/S phase showed colocalization of LANA with ORCs. Furthermore, replication of The TR-containing plasmid required both the N- and C termini of LANA in 293 and DG75 cells. Interestingly, our studies did not detect cellular ORCs associated with packaged viral DNA as an analysis of purified virions did not reveal the presence of ORCs, minichromosome maintenance proteins, or LANA. PMID- 16474133 TI - Inhibition of the ATM/p53 signal transduction pathway by Kaposi's sarcoma associated herpesvirus interferon regulatory factor 1. AB - Infected cells recognize viral replication as a DNA damage stress and elicit the ataxia telangiectasia-mutated (ATM)/p53-mediated DNA damage response signal transduction pathway as part of the host surveillance mechanisms, which ultimately induces the irreversible cell cycle arrest and apoptosis. Viruses have evolved a variety of mechanisms to counteract this host intracellular innate immunity. Kaposi's sarcoma-associated herpesvirus (KSHV) viral interferon regulatory factor 1 (vIRF1) interacts with the cellular p53 tumor suppressor through its central DNA binding domain, and this interaction inhibits transcriptional activation of p53. Here, we further demonstrate that KSHV vIRF1 downregulates the total p53 protein level by facilitating its proteasome-mediated degradation. Detailed biochemical study showed that vIRF1 interacted with cellular ATM kinase through its carboxyl-terminal transactivation domain and that this interaction blocked the activation of ATM kinase activity induced by DNA damage stress. As a consequence, vIRF1 expression greatly reduced the level of serine 15 phosphorylation of p53, resulting in an increase of p53 ubiquitination and thereby a decrease of its protein stability. These results indicate that KSHV vIRF1 comprehensively compromises an ATM/p53-mediated DNA damage response checkpoint by targeting both upstream ATM kinase and downstream p53 tumor suppressor, which might circumvent host growth surveillance and facilitate viral replication in infected cells. PMID- 16474134 TI - A single intranasal inoculation with a paramyxovirus-vectored vaccine protects guinea pigs against a lethal-dose Ebola virus challenge. AB - To determine whether intranasal inoculation with a paramyxovirus-vectored vaccine can induce protective immunity against Ebola virus (EV), recombinant human parainfluenza virus type 3 (HPIV3) was modified to express either the EV structural glycoprotein (GP) by itself (HPIV3/EboGP) or together with the EV nucleoprotein (NP) (HPIV3/EboGP-NP). Expression of EV GP by these recombinant viruses resulted in its efficient incorporation into virus particles and increased cytopathic effect in Vero cells. HPIV3/EboGP was 100-fold more efficiently neutralized by antibodies to EV than by antibodies to HPIV3. Guinea pigs infected with a single intranasal inoculation of 10(5.3) PFU of HPIV3/EboGP or HPIV3/EboGP-NP showed no apparent signs of disease yet developed a strong humoral response specific to the EV proteins. When these animals were challenged with an intraperitoneal injection of 10(3) PFU of EV, there were no outward signs of disease, no viremia or detectable EV antigen in the blood, and no evidence of infection in the spleen, liver, and lungs. In contrast, all of the control animals died or developed severe EV disease following challenge. The highly effective immunity achieved with a single vaccine dose suggests that intranasal immunization with live vectored vaccines based on recombinant respiratory viruses may be an advantageous approach to inducing protective responses against severe systemic infections, such as those caused by hemorrhagic fever agents. PMID- 16474135 TI - Intrahepatic hepatitis C virus replication correlates with chronic hepatitis C disease severity in vivo. AB - The role of viral factors in the pathogenesis of chronic hepatitis C is unknown. The objective of the present study was to characterize markers of hepatitis C virus (HCV) infection and replication in liver biopsy specimens obtained from 65 genotype 1-infected subjects, including 31 who were coinfected with human immunodeficiency virus (HIV), and to analyze associations between intrahepatic viral markers and hepatitis C disease severity. The percentages of liver cells harboring HCV genomes (%G) and replicative-intermediate RNAs (%RI) were evaluated using strand-specific in situ hybridization, while HCV core and NS3 antigens were assessed by immunocytochemistry. HIV-positive and HIV-negative subjects had similar mean grades and stages of liver disease and had similar indices of HCV infection and replication in liver, even though coinfected subjects had significantly shorter mean disease duration (P = 0.0003). Multivariate analysis showed that %G was not associated with grade or stage of liver disease (P = 0.5 and 0.4, respectively), while %RI was strongly associated with liver inflammation (P < 0.001), liver fibrosis (P < 0.001), and serum alanine aminotransferase levels (P = 0.01). NS3 antigen (but not core) was more frequently detected in HCV RI-positive versus RI-negative specimens (P = 0.028). These findings demonstrate a link between HCV proliferation and hepatitis C disease severity and suggest similar pathogenic mechanisms in HIV-positive and HIV-negative individuals. PMID- 16474136 TI - YRKL sequence of influenza virus M1 functions as the L domain motif and interacts with VPS28 and Cdc42. AB - Earlier studies have shown that the C-terminal half of helix 6 (H6) of the influenza A virus matrix protein (M1) containing the YRKL sequence is involved in virus budding (E. K.-W. Hui, S. Barman, T. Y. Yang, and D. P. Nayak, J. Virol. 77:7078-7092, 2003). In this report, we show that the YRKL sequence is the L domain motif of influenza virus. Like other L domains, YRKL can be inserted at different locations on the mutant M1 protein and can restore virus budding in a position-independent manner. Although YRKL is a part of the nuclear localization signal (NLS), the function of YRKL was independent of the NLS activity and the NLS function of M1 was not required for influenza virus replication. Some mutations in YRKL and the adjacent region caused a reduction in the virus titer by blocking virus release, and some affected virus morphology, producing elongated particles. Coimmunoprecipitation and Western blotting analyses showed that VPS28, a component of the ESCRT-I complex, and Cdc42, a member of the Rho family GTP-binding proteins, interacted with the M1 protein via the YRKL motif. In addition, depletion of VPS28 and Cdc42 by small interfering RNA resulted in reduction of influenza virus production. Moreover, overexpression of dominant negative Cdc42 inhibited influenza virus replication, whereas a constitutively active Cdc42 mutant enhanced virus production in infected cells. These results indicated that VPS28, a component of ESCRT-I, and Cdc42, a small G protein, are associated with the M1 protein and involved in the influenza virus life cycle. PMID- 16474137 TI - Structural characterization of the UL25 DNA-packaging protein from herpes simplex virus type 1. AB - Herpesviruses replicate their double stranded DNA genomes as high-molecular weight concatemers which are subsequently cleaved into unit-length genomes by a complex mechanism that is tightly coupled to DNA insertion into a preformed capsid structure, the procapsid. The herpes simplex virus type 1 UL25 protein is incorporated into the capsid during DNA packaging, and previous studies of a null mutant have demonstrated that its function is essential at the late stages of the head-filling process, either to allow packaging to proceed to completion or for retention of the viral genome within the capsid. We have expressed and purified an N-terminally truncated form of the 580-residue UL25 protein and have determined the crystallographic structure of the region corresponding to amino acids 134 to 580 at 2.1-Angstroms resolution. This structure, the first for any herpesvirus protein involved in processing and packaging of viral DNA, reveals a novel fold, a distinctive electrostatic distribution, and a unique "flexible" architecture in which numerous flexible loops emanate from a stable core. Evolutionary trace analysis of UL25 and its homologues in other herpesviruses was used to locate potentially important amino acids on the surface of the protein, leading to the identification of four putative docking regions for protein partners. PMID- 16474138 TI - Hierarchy among viral RNA (vRNA) segments in their role in vRNA incorporation into influenza A virions. AB - The genome of influenza A viruses comprises eight negative-strand RNA segments. Although all eight segments must be present in cells for efficient viral replication, the mechanism(s) by which these viral RNA (vRNA) segments are incorporated into virions is not fully understood. We recently found that sequences at both ends of the coding regions of the HA, NA, and NS vRNA segments of A/WSN/33 play important roles in the incorporation of these vRNAs into virions. In order to similarly identify the regions of the PB2, PB1, and PA vRNAs of this strain that are critical for their incorporation, we generated a series of mutant vRNAs that possessed the green fluorescent protein gene flanked by portions of the coding and noncoding regions of the respective segments. For all three polymerase segments, deletions at the ends of their coding regions decreased their virion incorporation efficiencies. More importantly, these regions not only affected the incorporation of the segment in which they reside, but were also important for the incorporation of other segments. This effect was most prominent with the PB2 vRNA. These findings suggest a hierarchy among vRNA segments for virion incorporation and may imply intersegment association of vRNAs during virus assembly. PMID- 16474139 TI - Glycosylation of the severe acute respiratory syndrome coronavirus triple spanning membrane proteins 3a and M. AB - The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein. The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses. It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other. The M protein plays a crucial role in coronavirus assembly and is glycosylated in all coronaviruses, either by N-linked or by O linked oligosaccharides. The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins. The proteins were expressed separately using the vaccinia virus T7 expression system, followed by metabolic labeling. Pulse-chase analysis showed that both proteins were modified, although in different ways. While the M protein acquired cotranslationally oligosaccharides that could be removed by PNGaseF, the 3a protein acquired its modifications posttranslationally, and they were not sensitive to the N glycosidase enzyme. The SARS-CoV 3a protein, however, was demonstrated to contain sialic acids, indicating the presence of oligosaccharides. O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants. In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars. Thus, the SARS-CoV 3a protein is an O glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated. PMID- 16474140 TI - Different de novo initiation strategies are used by influenza virus RNA polymerase on its cRNA and viral RNA promoters during viral RNA replication. AB - Various mechanisms are used by single-stranded RNA viruses to initiate and control their replication via the synthesis of replicative intermediates. In general, the same virus-encoded polymerase is responsible for both genome and antigenome strand synthesis from two different, although related promoters. Here we aimed to elucidate the mechanism of initiation of replication by influenza virus RNA polymerase and establish whether initiation of cRNA and viral RNA (vRNA) differed. To do this, two in vitro replication assays, which generated transcripts that had 5' triphosphate end groups characteristic of authentic replication products, were developed. Surprisingly, mutagenesis screening suggested that the polymerase initiated pppApG synthesis internally on the model cRNA promoter, whereas it initiated pppApG synthesis terminally on the model vRNA promoter. The internally synthesized pppApG could subsequently be used as a primer to realign, by base pairing, to the terminal residues of both the model cRNA and vRNA promoters. In vivo evidence, based on the correction of a mutated or deleted residue 1 of a cRNA chloramphenicol acetyltransferase reporter construct, supported this internal initiation and realignment model. Thus, influenza virus RNA polymerase uses different initiation strategies on its cRNA and vRNA promoters. To our knowledge, this is novel and has not previously been described for any viral RNA-dependent RNA polymerase. Such a mechanism may have evolved to maintain genome integrity and to control the level of replicative intermediates in infected cells. PMID- 16474141 TI - Distinct viral populations differentiate and evolve independently in a single perennial host plant. AB - The complex structure of virus populations has been the object of intensive study in bacteria, animals, and plants for over a decade. While it is clear that tremendous genetic diversity is rapidly generated during viral replication, the distribution of this diversity within a single host remains an obscure area in this field of science. Among animal viruses, only Human immunodeficiency virus and Hepatitis C virus populations have recently been thoroughly investigated at an intrahost level, where they are structured as metapopulations, demonstrating that the host cannot be considered simply as a "bag" containing a homogeneous or unstructured swarm of mutant viral genomes. In plants, a few reports suggested a possible heterogeneous distribution of virus variants at different locations within the host but provided no clues as to how this heterogeneity is structured. Here, we report the most exhaustive study of the structure and evolution of a virus population ever reported at the intrahost level through the analysis of a Prunus tree infected by Plum pox virus for over 13 years following a single inoculation event and by using analysis of molecular variance at different hierarchical levels combined with nested clade analysis. We demonstrate that, following systemic invasion of the host, the virus population differentiates into several distinct populations that are isolated in different branches, where they evolve independently through contiguous range expansion while colonizing newly formed organs. Moreover, we present and discuss evidence that the tree harbors a huge "bank" of viral clones, each isolated in one of the myriad leaves. PMID- 16474142 TI - A chromatin insulator-like element in the herpes simplex virus type 1 latency associated transcript region binds CCCTC-binding factor and displays enhancer blocking and silencing activities. AB - A previous study demonstrated that the latency-associated transcript (LAT) promoter and the LAT enhancer/reactivation critical region (rcr) are enriched in acetyl histone H3 (K9, K14) during herpes simplex virus type 1 (HSV-1) latency, whereas all lytic genes analyzed (ICP0, UL54, ICP4, and DNA polymerase) are not (N. J. Kubat, R. K. Tran, P. McAnany, and D. C. Bloom, J. Virol. 78:1139-1149, 2004). This suggests that the HSV-1 latent genome is organized into histone H3 (K9, K14) hyperacetylated and hypoacetylated regions corresponding to transcriptionally permissive and transcriptionally repressed chromatin domains, respectively. Such an organization implies that chromatin insulators, similar to those of cellular chromosomes, may separate distinct transcriptional domains of the HSV-1 latent genome. In the present study, we sought to identify cis elements that could partition the HSV-1 genome into distinct chromatin domains. Sequence analysis coupled with chromatin immunoprecipitation and luciferase reporter assays revealed that (i) the long and short repeats and the unique-short region of the HSV-1 genome contain clustered CTCF (CCCTC-binding factor) motifs, (ii) CTCF motif clusters similar to those in HSV-1 are conserved in other alphaherpesviruses, (iii) CTCF binds to these motifs on latent HSV-1 genomes in vivo, and (iv) a 1.5-kb region containing the CTCF motif cluster in the LAT region possesses insulator activities, specifically, enhancer blocking and silencing. The finding that CTCF, a cellular protein associated with chromatin insulators, binds to motifs on the latent genome and insulates the LAT enhancer suggests that CTCF may facilitate the formation of distinct chromatin boundaries during herpesvirus latency. PMID- 16474144 TI - Modeling and estimation of replication fitness of human immunodeficiency virus type 1 in vitro experiments by using a growth competition assay. AB - Growth competition assays have been developed to quantify the relative fitnesses of human immunodeficiency virus (HIV-1) mutants. In this article we develop mathematical models to describe viral/cellular dynamic interactions in the assay experiment, from which new competitive fitness indices or parameters are defined. These indices include the log fitness ratio (LFR), the log relative fitness (LRF), and the production rate ratio (PRR). From the population genetics perspective, we clarify the confusion and correct the inconsistency in the definition of relative fitness in the literature of HIV-1 viral fitness. The LFR and LRF are easier to estimate from the experimental data than the PRR, which was misleadingly defined as the relative fitness in recent HIV-1 research literature. Calculation and estimation methods based on two data points and multiple data points were proposed and were carefully studied. In particular, we suggest using both standard linear regression (method of least squares) and a measurement error model approach for more-accurate estimates of competitive fitness parameters from multiple data points. The developed methodologies are generally applicable to any growth competition assays. A user-friendly computational tool also has been developed and is publicly available on the World Wide Web at http://www.urmc.rochester.edu/bstools/vfitness/virusfitness.htm. PMID- 16474143 TI - Selective lymphocyte depletion during the early stage of the immune response to foot-and-mouth disease virus infection in swine. AB - Foot-and-mouth disease virus (FMDV) is the causative agent of a highly contagious vesicular disease of cloven-hoofed animals. In the present study we use FMDV serotype C infection of swine to determine, by analytical techniques, the direct ex vivo visualization of virus-infected immune cells during the first 17 days of infection. We report, for the first time, that FMDV C-S8c1 can infect T and B cells at short periods of time postinoculation, corresponding with the peak of the viremia. There is a significant lymphopenia that involves CD3(+) CD4(-) CD8(+/-), CD3(+) CD4(-) CD8(+)Tc, and CD3(+) CD4(+) CD8(+) memory Th but not CD3(+) CD4(+) CD8(-) naive Th lymphocytes. In addition, a profound depletion of the vast majority of peripheral T cells in lymph nodes and spleen is observed. This selective depletion of T cells is not due mainly to in situ death via apoptosis as visualized by the terminal deoxynucleotidyltransferase-mediated dUTP biotin nick end labeling (TUNEL) technique. Thus, early infection of T cells by FMDV may be the main cause of the observed T-cell depletion. Importantly, this lack of T cells is reflected in a reduced response to mitogen activation, which in many cases is totally eliminated. These data suggest a mechanism by which the virus causes a transient immunosuppression, subvert the immune systems, and spreads. These results have important implications for our understanding of early events in the development of a robust immune response against FMDV. PMID- 16474145 TI - Expression of Autographa californica multiple nucleopolyhedrovirus genes in mammalian cells and upregulation of the host beta-actin gene. AB - The gene expression of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) was examined in two types of mammalian cells, human HeLa14 and hamster BHK cells. DNA microarray analysis followed by reverse transcription-PCR identified at least 12 viral genes transcribed in both HeLa14 cells and BHK cells inoculated with AcMNPV. 5' rapid amplification of cDNA ends was carried out to examine the transcriptional fidelity of these genes in HeLa14 cells. The transcription of ie-1, ie-0 and gp64 was initiated at a baculovirus early gene motif, CAGT, accompanied by a TATA motif. In addition, the same splicing observed for ie-0 mRNA in Sf9 cells occurred in HeLa14 cells. While the transcription initiation sites for pe38 and p6.9 were not located in the CAGT motif, most of them were in a typical eukaryotic RNA polymerase II promoter structure (a conventional TATA motif and/or an initiator). Interestingly, the expression of beta-actin was upregulated in the mammalian cells inoculated with AcMNPV. Subsequent experiments using UV-inactivated virus confirmed the upregulation, suggesting that de novo synthesis of viral products is not required for the event. These results indicated that the AcMNPV genome acts as a template for transcription in mammalian cells through the usual infection pathway, though there is no evidence for the functional expression of viral genes at present. PMID- 16474146 TI - A single amino acid substitution in the West Nile virus nonstructural protein NS2A disables its ability to inhibit alpha/beta interferon induction and attenuates virus virulence in mice. AB - Alpha/beta interferons (IFN-alpha/beta) are key mediators of the innate immune response against viral infection. The ability of viruses to circumvent IFN alpha/beta responses plays a crucial role in determining the outcome of infection. In a previous study using subgenomic replicons of the Kunjin subtype of West Nile virus (WNV(KUN)), we demonstrated that the nonstructural protein NS2A is a major inhibitor of IFN-beta promoter-driven transcription and that a single amino acid substitution in NS2A (Ala30 to Pro [A30P]) dramatically reduced its inhibitory effect (W. J. Liu, H. B. Chen, X. J. Wang, H. Huang, and A. A. Khromykh, J. Virol. 78:12225-12235). Here we show that incorporation of the A30P mutation into the WNV(KUN) genome results in a mutant virus which elicits more rapid induction and higher levels of synthesis of IFN-alpha/beta in infected human A549 cells than that detected following wild-type WNV(KUN) infection. Consequently, replication of the WNV(KUN)NS2A/A30P mutant virus in these cells known to be high producers of IFN-alpha/beta was abortive. In contrast, both the mutant and the wild-type WNV(KUN) produced similar-size plaques and replicated with similar efficiency in BHK cells which are known to be deficient in IFN alpha/beta production. The mutant virus was highly attenuated in neuroinvasiveness and also attenuated in neurovirulence in 3-week-old mice. Surprisingly, the mutant virus was also partially attenuated in IFN alpha/betagamma receptor knockout mice, suggesting that the A30P mutation may also play a role in more efficient activation of other antiviral pathways in addition to the IFN response. Immunization of wild-type mice with the mutant virus resulted in induction of an antibody response of similar magnitude to that observed in mice immunized with wild-type WNV(KUN) and gave complete protection against challenge with a lethal dose of the highly virulent New York 99 strain of WNV. The results confirm and extend our previous original findings on the role of the flavivirus NS2A protein in inhibition of a host antiviral response and demonstrate that the targeted disabling of a viral mechanism for evading the IFN response can be applied to the development of live attenuated flavivirus vaccine candidates. PMID- 16474147 TI - A mutation in the human immunodeficiency virus type 1 Gag protein destabilizes the interaction of the envelope protein subunits gp120 and gp41. AB - The Gag protein of human immunodeficiency virus type 1 (HIV-1) associates with the envelope protein complex during virus assembly. The available evidence indicates that this interaction involves recognition of the gp41 cytoplasmic tail (CT) by the matrix protein (MA) region of Pr55(Gag). Here we show that substitution of Asp for Leu at position 49 (L49D) in MA results in a specific reduction in particle-associated gp120 without affecting the levels of gp41. Mutant virions were markedly reduced in single-cycle infectivity despite a relatively modest defect in fusion with target cells. Studies with HIV-1 particles containing decreased levels of envelope proteins suggested that the L49D mutation also inhibits a postentry step in infection. Truncation of the gp41 tail, or pseudotyping by vesicular stomatitis virus glycoprotein, restored both the fusion and infectivity of L49D mutant virions to wild-type levels. Truncation of gp41 also resulted in equivalent levels of gp120 on particles with and without the MA mutation and enhanced the replication of the L49D mutant virus in T cells. The impaired fusion and infectivity of L49D mutant particles were also complemented by a single point mutation in the gp41 CT that disrupted the tyrosine-containing endocytic motif. Our results suggest that an altered interaction between the MA domain of Gag and the gp41 cytoplasmic tail leads to dissociation of gp120 from gp41 during HIV-1 particle assembly, thus resulting in impaired fusion and infectivity. PMID- 16474148 TI - Association between hepatitis C virus and very-low-density lipoprotein (VLDL)/LDL analyzed in iodixanol density gradients. AB - Hepatitis C virus (HCV) RNA circulates in the blood of persistently infected patients in lipoviroparticles (LVPs), which are heterogeneous in density and associated with host lipoproteins and antibodies. The variability and lability of these virus-host complexes on fractionation has hindered our understanding of the structure of LVP and determination of the physicochemical properties of the HCV virion. In this study, HCV from an antibody-negative immunodeficient patient was analyzed using three fractionation techniques, NaBr gradients, isotonic iodixanol, and sucrose gradient centrifugation. Iodixanol gradients were shown to best preserve host lipoprotein-virus complexes, and all HCV RNA was found at densities below 1.13 g/ml, with the majority at low density, < or =1.08 g/ml. Immunoprecipitation with polyclonal antibodies against human ApoB and ApoE precipitated 91.8% and 95.0% of HCV with low density, respectively, suggesting that host lipoprotein is closely associated with HCV in a particle resembling VLDL. Immunoprecipitation with antibodies against glycoprotein E2 precipitated 25% of HCV with low density, providing evidence for the presence of E2 in LVPs. Treatment of serum with 0.5% deoxycholic acid in the absence of salt produced HCV with a density of 1.12 g/ml and a sedimentation coefficient of 215S. The diameters of these particles were calculated as 54 nm. Treatment of serum with 0.18% NP-40 produced HCV with a density of 1.18 g/ml, a sedimentation coefficient of 180S, and a diameter of 42 nm. Immunoprecipitation analysis showed that ApoB remained associated with HCV after treatment of serum with deoxycholic acid or NP 40, whereas ApoE was removed from HCV with these detergents. PMID- 16474149 TI - MicroRNA-guided processing impairs Plum pox virus replication, but the virus readily evolves to escape this silencing mechanism. AB - Since the discovery of microRNA (miRNA)-guided processing, a new type of RNA silencing, the possibility that such a mechanism could play a role in virus defense has been proposed. In this work, we have analyzed whether Plum pox virus (PPV) chimeras bearing miRNA target sequences (miR171, miR167, and miR159), which have been reported to be functional in Arabidopsis, were affected by miRNA function in three different host plants. Some of these PPV chimeras had clearly impaired infectivity compared with those carrying nonfunctional miRNA target sequences. The behaviors of PPV chimeras were similar but not identical in all the plants tested, and the deleterious effect on virus infectivity depended on the miRNA sequence cloned and on the site of insertion in the viral genome. The effect of the miRNA target sequence was drastically alleviated in transgenic plants expressing the silencing suppressor P1/HCPro. Furthermore, we show that virus chimeras readily escape RNA silencing interference through mutations within the miRNA target sequence, which mainly affected nucleotides matching the 5' terminal region of the miRNA. PMID- 16474150 TI - Potassium ion channels of Chlorella viruses cause rapid depolarization of host cells during infection. AB - Previous studies have established that chlorella viruses encode K(+) channels with different structural and functional properties. In the current study, we exploit the different sensitivities of these channels to Cs(+) to determine if the membrane depolarization observed during virus infection is caused by the activities of these channels. Infection of Chlorella NC64A with four viruses caused rapid membrane depolarization of similar amplitudes, but with different kinetics. Depolarization was fastest after infection with virus SC-1A (half time [t(1/2)], about 9 min) and slowest with virus NY-2A (t(1/2), about 12 min). Cs(+) inhibited membrane depolarization only in viruses that encode a Cs(+)-sensitive K(+) channel. Collectively, the results indicate that membrane depolarization is an early event in chlorella virus-host interactions and that it is correlated with viral-channel activity. This suggestion was supported by investigations of thin sections of Chlorella cells, which show that channel blockers inhibit virus DNA release into the host cell. Together, the data indicate that the channel is probably packaged in the virion, presumably in its internal membrane. We hypothesize that fusion of the virus internal membrane with the host plasma membrane results in an increase in K(+) conductance and membrane depolarization; this depolarization lowers the energy barrier for DNA release into the host. PMID- 16474151 TI - Ataxia-telangiectasia-mutated (ATM) protein can enhance human immunodeficiency virus type 1 replication by stimulating Rev function. AB - The ataxia-telangiectasia-mutated (ATM) kinase plays a central role in responses to various forms of DNA damage and has been suggested to facilitate human immunodeficiency virus type 1 (HIV-1) integration. Here, we describe a series of experiences that indicate that ATM can enhance HIV-1 replication by stimulating the action of the Rev viral posttranscriptional regulator. The Rev-dependent stimulation of viral late gene expression was observed with ATM-overexpressing cells, a result confirmed with a Rev-dependent reporter construct. Both parameters were also enhanced upon treatment of HeLa cells with caffeine, a xanthine that, in this cellular context, stimulates ATM activity. As well, decreased levels of virions with reduced infectivity were released by ATM knockdown cells. Notably, ATM overexpression did not stimulate the HIV-1 late gene expression within the context of Rev-independent constructs or the Rex dependent production of capsid from human T-cell leukemia virus type 1 proviral constructs. Altogether, these results indicate that ATM can positively influence HIV-1 Rev function. PMID- 16474152 TI - Host species-dependent population structure of a pollen-borne plant virus, Cherry leaf roll virus. AB - Cherry leaf roll virus (CLRV) belongs to the Nepovirus genus within the family Comoviridae. It has a host range which includes a number of wild tree and shrub species. The serological and molecular diversity of CLRV was assessed using a collection of isolates and samples recovered from woody and herbaceous host plants from different geographical origins. Molecular diversity was assessed by sequencing a short (375-bp) region of the 3' noncoding region (NCR) of the genomic RNAs while serological diversity was assessed using a panel of seven monoclonal antibodies raised initially against a walnut isolate of CLRV. The genomic region analyzed was shown to exhibit a significant degree of molecular variability with an average pairwise divergence of 8.5% (nucleotide identity). Similarly, serological variability proved to be high, with no single monoclonal antibody being able to recognize all isolates analyzed. Serological and molecular phylogenetic reconstructions showed a strong correlation. Remarkably, the diversity of CLRV populations is to a large extent defined by the host plant from which the viral samples are originally obtained. There are relatively few reports of plant viruses for which the genetic diversity is structured by the host plant. In the case of CLRV, we hypothesize that this situation may reflect the exclusive mode of transmission in natural plant populations by pollen and by seeds. These modes of transmission are likely to impose barriers to host change by the virus, leading to rapid biological and genetic separation of CLRV variants coevolving with different plant host species. PMID- 16474153 TI - Genetic association of the antiviral restriction factor TRIM5alpha with human immunodeficiency virus type 1 infection. AB - The innate antiviral factor TRIM5alpha restricts the replication of some retroviruses through its interaction with the viral capsid protein, leading to abortive infection. While overexpression of human TRIM5alpha results in modest restriction of human immunodeficiency virus type 1 (HIV-1), this inhibition is insufficient to block productive infection of human cells. We hypothesized that polymorphisms within TRIM5 may result in increased restriction of HIV-1 infection. We sequenced the TRIM5 gene (excluding exon 5) and the 4.8-kb 5' putative regulatory region in genomic DNA from 110 HIV-1-infected subjects and 96 exposed seronegative persons, along with targeted gene sequencing in a further 30 HIV-1-infected individuals. Forty-eight single nucleotide polymorphisms (SNPs), including 20 with allele frequencies of >1.0%, were identified. Among these were two synonymous and eight nonsynonymous coding polymorphisms. We observed no association between TRIM5 polymorphism in HIV-1-infected subjects and their set point viral load after acute infection, although one TRIM5 haplotype was weakly associated with more rapid CD4(+) T-cell loss. Importantly, a TRIM5 haplotype containing the nonsynonymous SNP R136Q showed increased frequency among HIV-1 infected subjects relative to exposed seronegative persons, with an odds ratio of 5.49 (95% confidence interval = 1.83 to 16.45; P = 0.002). Nonetheless, we observed no effect of individual TRIM5alpha nonsynonymous mutations on the in vitro HIV-1 susceptibility of CD4(+) T cells. Therefore, any effect of TRIM5alpha polymorphism on HIV-1 infection in primary lymphocytes may depend on combinations of SNPs or on DNA sequences in linkage disequilibrium with the TRIM5alpha coding sequence. PMID- 16474154 TI - Extensive recombination among human immunodeficiency virus type 1 quasispecies makes an important contribution to viral diversity in individual patients. AB - Although recombination during human immunodeficiency virus type 1 (HIV-1) replication in vitro and in vivo has been documented, little information is available concerning the extent that recombination contributes to the diversity of HIV-1 quasispecies in the course of infection in individual patents. To investigate the impact of recombination on viral diversity, we developed a technique that permits the isolation of contemporaneous clonal viral populations resulting from single infectious events by plasma-derived viruses, thereby permitting the assessment of recombination throughout the viral genomes, including widely separated loci, from individual patients. A comparison of the genomic sequences of clonal viruses from six patients, including patients failing treatment with antiretroviral therapy, demonstrated strong evidence for extensive recombination. Recombination increased viral diversity through two distinct mechanisms. First, evolutionary bottlenecks appeared to be restricted to minimal segments of the genome required to obtain selective advantage, thereby preserving diversity in adjacent regions. Second, recombination between adjacent gene segments appeared to generate diversity in both pol and env genes. Thus, the shuffling of resistance mutations within the genes coding for the protease and reverse transcriptase, as well as recombination between these regions, could increase the diversity of drug resistance genotypes. These findings demonstrate that recombination in HIV-1 contributes to the diversity of viral quasispecies by restricting evolutionary bottlenecks to gene segments and by generating novel genotypes in pol and env, supporting the idea that recombination may be critical to adaptive evolution of HIV in the face of constantly moving selective pressures, whether exerted by the immune system or antiretroviral therapy. PMID- 16474155 TI - Pausing during reverse transcription increases the rate of retroviral recombination. AB - Retroviruses package two copies of genomic RNA into viral particles. During the minus-sense DNA synthesis step of reverse transcription, the nascent DNA can transfer multiple times between the two copies of the genome, resulting in recombination. The mechanism for this process is similar to the process of obligate strand transfers mediated by the repeat and primer binding site sequences. The location at which the DNA 3' terminus completely transfers to the second RNA strand defines the point of crossover. Previous work in vitro demonstrated that reverse transcriptase pausing has a significant impact on the location of the crossover, with a proportion of complete transfer events occurring very close to pause sites. The role of pausing in vivo, however, is not clearly understood. By employing a murine leukemia virus-based single-cycle infection assay, strong pausing was shown to increase the probability of recombination, as reflected in the reconstitution of green fluorescent protein expression. The infection assay results were directly correlated with the presence of strong pause sites in reverse transcriptase primer extension assays in vitro. Conversely, when pausing was diminished in vitro, without changing the sequence of the RNA template involved in recombination, there was a significant reduction in recombination in vivo. Together, these data demonstrate that reverse transcriptase pausing, as observed in vitro, directly correlates with recombination during minus-sense DNA synthesis in vivo. PMID- 16474156 TI - A novel alternative splicing isoform of human T-cell leukemia virus type 1 bZIP factor (HBZ-SI) targets distinct subnuclear localization. AB - Adult T-cell leukemia (ATL) is associated with prior infection with human T-cell leukemia virus type 1 (HTLV-1); however, the mechanism by which HTLV-1 causes adult T-cell leukemia has not been fully elucidated. Recently, a functional basic leucine zipper (bZIP) protein coded in the minus strand of HTLV-1 genome (HBZ) was identified. We report here a novel isoform of the HTLV-1 bZIP factor (HBZ), HBZ-SI, identified by means of reverse transcription-PCR (RT-PCR) in conjunction with 5' and 3' rapid amplification of cDNA ends (RACE). HBZ-SI is a 206-amino acid-long protein and is generated by alternative splicing between part of the HBZ gene and a novel exon located in the 3' long terminal repeat of the HTLV-1 genome. Consequently, these isoforms share >95% amino acid sequence identity, and differ only at their N termini, indicating that HBZ-SI is also a functional protein. Duplex RT-PCR and real-time quantitative RT-PCR analyses showed that the mRNAs of these isoforms were expressed at equivalent levels in all ATL cell samples examined. Nonetheless, we found by Western blotting that the HBZ-SI protein was preferentially expressed in some ATL cell lines examined. A key finding was obtained from the subcellular localization analyses of these isoforms. Despite their high sequence similarity, each isoform was targeted to distinguishable subnuclear structures. These data show the presence of a novel isoform of HBZ in ATL cells, and in addition, shed new light on the possibility that each isoform may play a unique role in distinct regions in the cell nucleus. PMID- 16474157 TI - Preferential infection of mature dendritic cells by mouse hepatitis virus strain JHM. AB - Mouse hepatitis virus strain JHM (MHV-JHM) causes acute encephalitis and acute and chronic demyelinating diseases in mice. Dendritic cells (DCs) are key cells in the initiation of innate and adaptive immune responses, and infection of these cells could potentially contribute to a dysregulated immune response; consistent with this, recent results suggest that DCs are readily infected by another strain of mouse hepatitis virus, the A59 strain (MHV-A59). Herein, we show that the JHM strain also productively infected DCs. Moreover, mature DCs were at least 10 times more susceptible than immature DCs to infection with MHV-JHM. DC function was impaired after MHV-JHM infection, resulting in decreased stimulation of CD8 T cells in vitro. Preferential infection of mature DCs was not due to differential expression of the MHV-JHM receptor CEACAM-1a on mature or immature cells or to differences in apoptosis. Although we could not detect infected DCs in vivo, both CD8(+) and CD11b(+) splenic DCs were susceptible to infection with MHV-JHM directly ex vivo. This preferential infection of mature DCs may inhibit the development of an efficient immune response to the virus. PMID- 16474158 TI - Nature of nonfunctional envelope proteins on the surface of human immunodeficiency virus type 1. AB - Human immunodeficiency virus type 1 (HIV-1) neutralizing antibodies are thought be distinguished from nonneutralizing antibodies by their ability to recognize functional gp120/gp41 envelope glycoprotein (Env) trimers. The antibody responses induced by natural HIV-1 infection or by vaccine candidates tested to date consist largely of nonneutralizing antibodies. One might have expected a more vigorous neutralizing response, particularly against virus particles that bear functional trimers. The recent surprising observation that nonneutralizing antibodies can specifically capture HIV-1 may provide a clue relating to this paradox. Specifically, it was suggested that forms of Env, to which nonneutralizing antibodies can bind, exist on virus surfaces. Here, we present evidence that HIV-1 particles bear nonfunctional gp120/gp41 monomers and gp120 depleted gp41 stumps. Using a native electrophoresis band shift assay, we show that antibody-trimer binding predicts neutralization and that the nonfunctional forms of Env may account for virus capture by nonneutralizing antibodies. We hypothesize that these nonfunctional forms of Env on particle surfaces serve to divert the antibody response, helping the virus to evade neutralization. PMID- 16474159 TI - Natural killer cell function is well preserved in asymptomatic human immunodeficiency virus type 2 (HIV-2) infection but similar to that of HIV-1 infection when CD4 T-cell counts fall. AB - Natural killer (NK) cells are potent effectors of natural immunity and their activity prevents human immunodeficiency virus type 1 (HIV-1) viral entry and viral replication. We sought to determine whether NK immune responses are associated with different clinical course of HIV-1 and HIV-2 infections. A cross sectional analysis of NK cell responses was undertaken in 30 HIV-1 and 30 HIV-2 subjects in each of three categories of CD4(+)-T-cell counts (>500, 200 to 500, and <200 cells/microl) and in 50 HIV-uninfected control subjects. Lytic activity and gamma interferon (IFN-gamma) secretion were measured by chromium release and enzyme-linked immunospot assays, respectively. Flow cytometry was used to assess intracellular cytokines and chemokines. Levels of NK cytotoxicity were significantly higher in HIV-2 than in HIV-1 infections in subjects with high CD4(+)-T-cell counts and were similar to that of the healthy controls. In these HIV-2 subjects, cytolytic activity was positively correlated to NK cell count and inversely related to plasma viremia. Levels of intracellular MIP-1beta, RANTES, tumor necrosis factor alpha, and IFN-gamma produced by NK CD56(bright) cells were significantly higher in HIV-2- than HIV-1-infected subjects with high CD4(+)-T cell counts but fell to similar levels as CD4 counts dropped. The data suggest efficient cytolytic and chemokine-suppressive activity of NK cells early in HIV-2 infection, which is associated with high CD4(+) T-cell counts. Enhancement of these functions may be important in immune-based therapy to control HIV disease. PMID- 16474160 TI - Effect of epitope position on neutralization by anti-human immunodeficiency virus monoclonal antibody 2F5. AB - The membrane-proximal region of the human immunodeficiency virus type 1 (HIV-1) transmembrane protein (TM) is critical for envelope (Env)-mediated membrane fusion and contains the target for broadly reactive neutralizing antibody 2F5. It has been proposed that 2F5 neutralization might involve interaction of its long, hydrophobic, complementarity-determining region (CDR) H3, with adjacent viral membrane. Using Moloney murine leukemia virus (MLV) as a tool, we examined the effect of epitope position on 2F5 neutralization. When the 2F5 epitope was inserted in the proline-rich region of MLV Env surface protein (SU), 2F5 blocked cell fusion and virus infection, whereas MLV with a hemagglutinin (HA) epitope at the same position was not neutralized by anti-HA, even though the antibodies bound their respective Envs on the surface of infected cells and viruses equally well. When the 2F5 epitope was inserted in the MLV Env TM at a position comparable to its natural position in HIV-1 TM, 2F5 antibody blocked Env-mediated cell fusion. Epitope position had subtle effects on neutralization by 2F5: the antibody concentration for 50% inhibition of cell fusion was more than 10-fold lower when the 2F5 epitope was in SU than in TM, and inhibition was less complete at high concentrations of antibody; we discuss possible explanations for these effects of epitope position. Since membrane proximity was not required for neutralization by 2F5 antibody, we speculate that the CDR H3 of 2F5 contributes to neutralization by destabilizing an adjacent protein rather than by inserting into an adjacent membrane. PMID- 16474162 TI - Crucial role of the 5' conserved structure of bamboo mosaic virus satellite RNA in downregulation of helper viral RNA replication. AB - Satellite RNA of Bamboo mosaic virus (satBaMV), a single-stranded mRNA type satellite encoding a protein of 20 kDa (P20), depends on the helper BaMV for replication and encapsidation. Two satBaMV isolates, BSF4 and BSL6, exhibit distinctly differential phenotypes in Nicotiana benthamiana plants when coinoculated with BaMV RNA. BSL6 significantly reduces BaMV RNA replication and suppresses the BaMV-induced symptoms, whereas BSF4 does not. By studies with chimeric satBaMVs generated by exchanging the components between BSF4 and BSL6, the genetic determinants responsible for the downregulation of BaMV replication and symptom expression were mapped at the 5' untranslated region (UTR) of BSL6. The 5' UTR of BSL6 alone is sufficient to diminish BaMV RNA replication when the 5' UTR is inserted in cis into the BaMV expression vector or when coinoculation with mutants that block the synthesis of P20 protein takes place. Further, the 5' UTR of natural satBaMV isolates contains one hypervariable (HV) region which folds into a conserved apical hairpin stem-loop (AHSL) structure (W. B. Yeh, Y. H. Hsu, H. C. Chen, and N. S. Lin, Virology 330:105-115, 2004). Interchanges of AHSL segment of HV regions between BSF4 and BSL6 led to the ability of chimeric satBaMV to interfere with BaMV replication and symptom expression. The conserved secondary structure within the HV region is a potent determinant of the downregulation of helper virus replication. PMID- 16474163 TI - Human herpesvirus 1 UL24 gene encodes a potential PD-(D/E)XK endonuclease. AB - Using Meta-BASIC, a highly sensitive method for detection of distant similarity between proteins, we have identified another potential PD-(D/E)XK endonuclease in human herpesvirus 1 (HHV-1) encoded by the UL24 gene. The universal presence of UL24 in completed herpesviral genomes of three major subfamilies, Alphaherpesvirinae, Betaherpesvirinae, and Gammaherpesvirinae, suggests a fundamental role for this predicted PD-(D/E)XK endonuclease activity in the viral life cycle. PMID- 16474161 TI - Virus and cell RNAs expressed during Epstein-Barr virus replication. AB - Changes in Epstein-Barr virus (EBV) and cell RNA levels were assayed following immunoglobulin G (IgG) cross-linking-induced replication in latency 1-infected Akata Burkitt B lymphoblasts. EBV replication as assayed by membrane gp350 expression was approximately 5% before IgG cross-linking and increased to more than 50% 48 h after induction. Seventy-two hours after IgG cross-linking, gp350 positive cells excluded propidium iodide as well as gp350-negative cells. EBV RNA levels changed temporally in parallel with previously defined sensitivity to inhibitors of protein or viral DNA synthesis. BZLF1 immediate-early RNA levels doubled by 2 h and reached a peak at 4 h, whereas BMLF1 doubled by 4 h with a peak at 8 h, and BRLF1 doubled by 8 h with peak at 12 h. Early RNAs peaked at 8 to 12 h, and late RNAs peaked at 24 h. Hybridization to intergenic sequences resulted in evidence for new EBV RNAs. Surprisingly, latency III (LTIII) RNAs for LMP1, LMP2, EBNALP, EBNA2, EBNA3A, EBNA3C, and BARTs were detected at 8 to 12 h and reached maxima at 24 to 48 h. EBNA2 and LMP1 were at full LTIII levels by 48 h and localized to gp350-positive cells. Thus, LTIII expression is a characteristic of late EBV replication in both B lymphoblasts and epithelial cells in immune-comprised people (J. Webster-Cyriaque, J. Middeldorp, and N. Raab Traub, J. Virol. 74:7610-7618, 2000). EBV replication significantly altered levels of 401 Akata cell RNAs, of which 122 RNAs changed twofold or more relative to uninfected Akata cells. Mitogen-activated protein kinase levels were significantly affected. Late expression of LTIII was associated with induction of NF-kappaB responsive genes including IkappaBalpha and A20. The exclusion of propidium, expression of EBV LTIII RNAs and proteins, and up-regulation of specific cell RNAs are indicative of vital cell function late in EBV replication. PMID- 16474164 TI - The level of reverse transcriptase (RT) in human immunodeficiency virus type 1 particles affects susceptibility to nonnucleoside RT inhibitors but not to lamivudine. AB - We investigated the relationship between the level of reverse transcriptase (RT) in human immunodeficiency virus type 1 (HIV-1) particles and susceptibility to nonnucleoside reverse transcriptase inhibitors (NNRTIs). HIV-1 virions containing different active levels of RT were generated. Susceptibility to the NNRTIs efavirenz and nevirapine was inversely proportional to the level of enzymatically active RT. However, the sensitivity of HIV-1 to the nucleoside analog 3TC was not affected by the level of RT per particle. These data indicate that the susceptibility of HIV-1 to NNRTIs is influenced by RT activity. PMID- 16474165 TI - Herpes simplex virus tegument protein VP16 is a component of primary enveloped virions. AB - Immunogold electron microscopy was used to determine whether the tegument proteins VP13/14, VP22, and VP16 of herpes simplex virus type 1 (HSV1) are components of primary enveloped virions. Whereas VP13/14 and VP22 were not detected in virus particles in the perinuclear space and were present in only mature extracellular virions, VP16 was acquired prior to primary envelopment of the virus at the inner nuclear membrane. This finding highlights potential similarities and differences between HSV1 and the related alphaherpesvirus, pseudorabies virus, in which the homologues of all three of these tegument proteins are not incorporated into the virion until secondary envelopment. PMID- 16474166 TI - Hypoxia regulates cross-talk between Syk and Lck leading to breast cancer progression and angiogenesis. AB - Hypoxia is a key parameter that controls tumor angiogenesis and malignant progression by regulating the expression of several oncogenic molecules. The nonreceptor protein-tyrosine kinases Syk and Lck play crucial roles in the signaling mechanism of various cellular processes. The enhanced expression of Syk in normal breast tissue but not in malignant breast carcinoma has prompted us to investigate its potential role in mammary carcinogenesis. Accordingly, we hypothesized that hypoxia/reoxygenation (H/R) may play an important role in regulating Syk activation, and Lck may be involved in this process. In this study, we have demonstrated that H/R differentially regulates Syk phosphorylation and its subsequent interaction and cross-talk with Lck in MCF-7 cells. Moreover, Syk and Lck play differential roles in regulating Sp1 activation and expressions of melanoma cell adhesion molecule (MelCAM), urokinase-type plasminogen activator (uPA), matrix metalloproteinase-9 (MMP-9), and vascular endothelial growth factor (VEGF) in response to H/R. Overexpression of wild type Syk inhibited the H/R induced uPA, MMP-9, and VEGF expression but up-regulated MelCAM expression. Our data also indicated that MelCAM acts as a tumor suppressor by negatively regulating H/R-induced uPA secretion and MMP-9 activation. The mice xenograft study showed the cross-talk between Syk and Lck regulated H/R-induced breast tumor progression and further correlated with the expressions of MelCAM, uPA, MMP 9, and VEGF. Human clinical specimen analysis supported the in vitro and in vivo findings. To our knowledge, this is first report that the cross-talk between Syk and Lck regulates H/R-induced breast cancer progression and further suggests that Syk may act as potential therapeutic target for the treatment of breast cancer. PMID- 16474167 TI - The WD40 repeats of FANCL are required for Fanconi anemia core complex assembly. AB - Fanconi anemia (FA) is an autosomal recessive disorder characterized by aplastic anemia, cancer susceptibility, and cellular sensitivity to mitomycin C. Eight of the 11 cloned Fanconi anemia gene products (FANCA, -B, -C, -E, -F, -G, -L, and M) form a multisubunit nuclear complex (FA core complex) required for monoubiquitination of a downstream FA protein, FANCD2. FANCL, which possesses three WD40 repeats and a plant homeodomain (PHD), is the putative E3 ubiquitin ligase subunit of the FA complex. Here, we demonstrate that the WD40 repeats of FANCL are required for interaction with other subunits of the FA complex. The PHD is dispensable for this interaction, although it is required for FANCD2 mono ubiquitination. The PHD of FANCL also shares sequence similarity to the canonical RING finger of c-CBL, including a conserved tryptophan required for E2 binding by c-CBL. Mutation of this tryptophan in the FANCL PHD significantly impairs in vivo mono-ubiquitination of FANCD2 and in vitro auto-ubiquitination activity, and partially impairs restoration of mitomycin C resistance. We propose a model in which FANCL, via its WD40 region, binds the FA complex and, via its PHD, recruits an as-yet-unidentified E2 for mono-ubiquitination of FANCD2. PMID- 16474168 TI - Stage matters: choosing relevant model systems to address hypotheses in diet and cancer chemoprevention research. AB - Clinical evidence reveals that the efficacy of dietary factors to prevent cancer is probably stage-dependent. The ability to demonstrate stage-specific effects of dietary compounds on normal, preneoplastic and malignant cell models may provide insights into puzzling clinical results from cancer chemoprevention trials. The relevance of these models to the field of cancer prevention is immense and will undoubtedly facilitate the ability to discover which dietary factors are most effective at preventing cancer and which, if any, specific steps in neoplastic transformation render cells refractory to the effects of dietary compounds. There are illustrative examples where exposure of high-risk individuals to dietary chemopreventive agents increases rather than decreases cancer risk. While geneticists and clinical oncologists acknowledge the morphological continuum along which tumors develop in specific tissues, tumor cells, rather than normal and preneoplastic cells, continue to be the primary in vitro reductionist tool employed to elucidate mechanisms underlying disease progression and to investigate the potential utility of dietary as well as other chemopreventive agents. Currently, there are few relevant model systems to study the progression of neoplastic transformation, especially in epithelial cells. We highlight examples of model systems isolated from prostate, breast, endometrial and intestinal tissue, with special emphasis on a specific set of non-tumorigenic, conditionally immortal cell lines derived from C57/BL6 mice [YAMC (Young Adult Mouse Colon cells; Apc+/+) cells and IMCE (Immorto-Min Colonic Epithelium cells; ApcMin/+) cells] that have yielded important information on early events in colorectal neoplasia development. These cell lines are an illustrative example of how researchers can examine stage-dependent effects of specific dietary components on carcinogenesis. The utilization of cell culture systems modeling early, middle and late stages of tumorigenesis will yield important insights into mechanisms by which dietary components impact cancer progression. PMID- 16474169 TI - Matrix metalloproteinase-7 increases resistance to Fas-mediated apoptosis and is a poor prognostic factor of patients with colorectal carcinoma. AB - The ability of tumor cells to resist apoptosis triggered by immune cells results in their escape from immune surveillance of the host. A critical effector of apoptosis is the Fas/Fas ligand (FasL) system that mediates the tumoricidal effects of cytotoxic T cells. Recently, in vitro cleavage of Fas expressed in various tumor cells by matrix metalloproteinase-7 (MMP-7) was demonstrated. In the present study, we first analyzed the influence of this metalloproteinase on Fas signaling in SW480, HCT-15 and HT-29 colorectal carcinoma (CRC) cells by assessing their responses to either an agonistic Fas antibody (CH11) or the FasL bearing Jurkat cells after they were pretreated with MMP-7. Interestingly, both antibody- and Jurkat cell-induced apoptosis in three different CRC lines were significantly reduced by MMP-7 pretreatment. Additionally, immunohistochemical (IHC) staining was used to examine the expression levels of MMP-7 and Fas in tumor samples of 54 CRC patients. In agreement with our in vitro observation, the expression of MMP-7 in tumor tissues was inversely correlated with those of Fas (P < 0.001; chi2-test). Moreover, shortened survival was found in patients with a higher MMP-7 and a lower Fas expression, respectively, in their tumor tissues (P < 0.0001). Finally, by multivariate analysis, we discovered that MMP-7 (P = 0.001) and Fas levels (P = 0.036) were independent prognostic factors for CRC patients. These results suggest that Fas downregulation and a consequential increased resistance to FasL-triggered apoptosis resulting from upregulated MMP-7 in colorectal cancer cells could be a key mechanism for their escape from the immune surveillance, thereby predicting a poor survival in CRC patients. PMID- 16474170 TI - Fractionation of grape seed extract and identification of gallic acid as one of the major active constituents causing growth inhibition and apoptotic death of DU145 human prostate carcinoma cells. AB - The anti-cancer efficacy of grape seed extract (GSE) against prostate cancer (PCA) via its anti-proliferative, pro-apoptotic and anti-angiogenic activities in both cell culture and animal models have recently been described by us. GSE is a complex mixture containing gallic acid (GA), catechin (C), epicatechin (EC) and several oligomers (procyanidins) of C and/or EC, some of which are esterified to GA. To determine which components are most active against PCA, an ethyl acetate extract of GSE was separated by reverse-phase high-performance liquid chromatography (HPLC) into three fractions. Fraction 1 was far more effective than others in causing growth inhibition and apoptotic death of human PCA DU145 cells. Of the components in this fraction, GA showed a very strong dose- and time dependent growth inhibition and apoptotic death of DU145 cells, but C and procyanidins B1 (EC-C dimer), B2 (EC-EC dimer) and B3 (C-C dimer) were nearly ineffective. Mechanistic studies demonstrated a strong caspase-9, caspase-3 and poly (ADP-ribose) polymerase (PARP) cleavages by GA in DU145 cells. Procyanidin oligomers eluting in HPLC Fractions 2 and 3 were obtained in larger quantities by separating GSE into eight fractions (I-VIII) on a gel filtration column. All fractions were analyzed by HPLC-UV and negative-ion electrospray mass spectrometry. Fractions I-III contained the active compound GA and inactive components C, EC, B1 and B2. Fraction IV contained other dimers and a dimer-GA ester and was also less active than GSE in DU145 cells. Fractions V-VIII, however, caused significant growth inhibition and apoptosis with the highest activity present in the later fractions that contained procyanidin trimers and GA esters of dimers and trimers. Together, these observations identify GA as one of the major active constituents in GSE. Several procyanidins, however, and especially the gallate esters of dimers and trimers also may be efficacious against PCA and merit further investigation. PMID- 16474171 TI - Convergent transcriptional profiles induced by endogenous estrogen and distinct xenoestrogens in breast cancer cells. AB - Estrogen receptors display high levels of promiscuity in accommodating a wide range of ligand structures, but the functional consequence of changing receptor conformations in complex with distinct agonists is highly controversial. To determine variations in the transactivation capacity induced by different estrogenic agonists, we assessed global transcriptional profiles elicited by natural or synthetic xenoestrogens in comparison with the endogenous hormone 17beta-estradiol. Human MCF7 and T47D carcinoma cells, representing the most frequently used model systems for tumorigenic responses in the mammary gland, were synchronized by hormone starvation during 48 h. Subsequently, a 24 h exposure was carried out with equipotent concentrations of the selected xenoestrogens or 17beta-estradiol. Analysis of messenger RNA was performed on high-density oligonucleotide microarrays that display the sequences of 33,000 human transcripts, yielding a total of 181 gene products that are regulated upon estrogenic stimulation. Surprisingly, genistein (a phytoestrogen), bisphenol-A and polychlorinated biphenyl congener 54 (two synthetic xenoestrogens) produced highly congruent genomic fingerprints by regulating the same range of human genes. Also, the monotonous genomic signature observed in response to xenoestrogens is identical to the transcriptional effects induced by physiological concentrations of 17beta-estradiol. This striking functional convergence indicates that the transcription machinery is largely insensitive to the particular structure of estrogen receptor agonists. The occurrence of such converging transcriptional programs reinforces the hypothesis that multiple xenoestrogenic contaminants, of natural or anthropogenic origin, may act in conjunction with the endogenous hormone to induce additive effects in target tissues. PMID- 16474172 TI - TagSNP analyses of the PON gene cluster: effects on PON1 activity, LDL oxidative susceptibility, and vascular disease. AB - Paraoxonase 1 (PON1) activity is consistently predictive of vascular disease, although the genotype at four functional PON1 polymorphisms is not. To address this inconsistency, we investigated the role of all common PON1 genetic variability, as measured by tagging single-nucleotide polymorphisms (tagSNPs), in predicting PON1 activity for phenylacetate hydrolysis, LDL susceptibility to oxidation ex vivo, plasma homocysteine (Hcy) levels, and carotid artery disease (CAAD) status. The biological goal was to establish whether additional common genetic variation beyond consideration of the four known functional SNPs improves prediction of these phenotypes. PON2 and PON3 tagSNPs were secondarily evaluated. Expanded analysis of an additional 26 tagSNPs found evidence of previously undescribed common PON1 polymorphisms that affect PON1 activity independently of the four known functional SNPs. PON1 activity was not significantly correlated with LDL oxidative susceptibility, but genotypes at the PON1(-108) promoter polymorphism and several other PON1 SNPs were. Neither PON1 activity nor PON1 genotype was significantly correlated with plasma Hcy levels. This study revealed previously undetected common functional PON1 polymorphisms that explain 4% of PON1 activity and a high rate of recombination in PON1, but the sum of the common PON1 locus variation does not explain the relationship between PON1 activity and CAAD. PMID- 16474173 TI - Lipophorin receptor of Bombyx mori: cDNA cloning, genomic structure, alternative splicing, and isolation of a new isoform. AB - The cDNA and genomic structure of a putative lipophorin receptor from the silkworm, Bombyx mori (BmLpR), indicated the presence of four isoforms, designated LpR1, LpR2, LpR3, and LpR4. The deduced amino acid sequence of each isoform showed five functional domains that are homologous to vertebrate very low density lipoprotein receptor (VLDLR). All four isoforms seem to have originated from a single gene by alternative splicing and were differentially expressed in a tissue- and stage-specific manner. BmLpR1 harbored an additional 27 amino acids in the O-linked sugar domain, resulting in an extra exon. The silkworm BmLpR gene consisted of 16 exons separated by 15 introns spanning >122 kb and was at least three times larger than the human VLDLR gene. Surprisingly, one of the isoforms, LpR4, was expressed specifically in the brain and central nervous system. Additionally, it had a unique cytoplasmic tail, leading to the proposition that it represents a new candidate LpR for possible brain-related function(s). This is the first report on the genomic characterization of an arthropod lipoprotein receptor gene and the identification of a brain-specific receptor variant from a core member of the low density lipoprotein receptor family in invertebrates. PMID- 16474174 TI - Variants at the APOA5 locus, association with carotid atherosclerosis, and modification by obesity: the Framingham Study. AB - Genetic variation at the apolipoprotein A5 gene (APOA5) is associated with increased triglyceride concentrations, a risk factor for atherosclerosis. Carotid intimal medial thickness (IMT) is a surrogate measure of atherosclerosis burden. We sought to determine the association of common APOA5 genetic variants with carotid IMT and stenosis. A total of 2,273 Framingham Offspring Study participants underwent carotid ultrasound and had data on at least one of the five APOA5 variants (-1131T>C, -3A>G, 56C>G, IVS3+476G >A, and 1259T>C). Although none of the individual variants was significantly associated with carotid measures, the haplotype defined by the presence of the rare allele of the 56C>G variant was associated with a higher common carotid artery (CCA) IMT compared with the wild-type haplotype (0.75 vs. 0.73 mm; P < 0.05). The rare allele of each of the -1131T >C, -3A>G, IVS3+476G>A, and 1259T>C variants and the haplotype defined by the presence of the rare alleles in these four variants were each significantly associated with CCA IMT in obese participants. These associations remained significant even after adjustment for triglycerides. APOA5 variants were associated with CCA IMT, particularly in obese participants. The mechanism of these associations and the effect modification by obesity are independent of fasting triglyceride levels. PMID- 16474175 TI - Proteomic approach to identification of proteins reactive for abasic sites in DNA. AB - Apurinic/apyrimidinic (AP) sites, a prominent type of DNA damage, are repaired through the base excision repair mechanism in both prokaryotes and eukaryotes and may interfere with many other cellular processes. A full repertoire of AP site binding proteins in cells is presently unknown, preventing reliable assessment of harm inflicted by these ubiquitous lesions and of their involvement in the flux of DNA metabolism. We present a proteomics-based strategy for assembling at least a partial catalogue of proteins capable of binding AP sites in DNA. The general scheme relies on the sensitivity of many AP site-bound protein species to NaBH(4) cross-linking. An affinity-tagged substrate is used to facilitate isolation of the cross-linked species, which are then separated and analyzed by mass spectrometry methods. We report identification of seven proteins from Escherichia coli (AroF, DnaK, MutM, PolA, TnaA, TufA, and UvrA) and two proteins from bakers' yeast (ARC1 and Ygl245wp) reactive for AP sites in this system. PMID- 16474176 TI - RAD50 and NBS1 are breast cancer susceptibility genes associated with genomic instability. AB - The Mre11 complex, composed of RAD50, NBS1 and MRE11, has an essential role in the maintenance of genomic integrity and preventing cells from malignancy. Here we report the association of three Mre11 complex mutations with hereditary breast cancer susceptibility, studied by using a case-control design with 317 consecutive, newly diagnosed Northern Finnish breast cancer patients and 1000 geographically matched healthy controls (P = 0.0004). RAD50 687delT displayed significantly elevated frequency in the studied patients (8 out of 317, OR 4.3, 95% CI 1.5-12.5, P= 0.008), which indicates that it is a relatively common low penetrance risk allele in this cohort. Haplotype analysis and the screening of altogether 512 additional breast cancer cases from Sweden, Norway and Iceland suggest that RAD50 687delT is a Finnish founder mutation, not present in the other Nordic cohorts. The RAD50 IVS3-1G>A splicing mutation leading to translational frameshift was observed in one patient, and the NBS1 Leu150Phe missense mutation affecting a conserved residue in the functionally important BRCA1 carboxy-terminal (BRCT) domain in two patients, both being absent from 1000 controls. Microsatellite marker analysis showed that loss of the wild-type allele was not involved in the tumorigenesis in any of the studied mutation carriers, but they all showed increased genomic instability assessed by cytogenetic analysis of peripheral blood T-lymphocytes (P = 0.006). In particular, the total number of chromosomal rearrangements was significantly increased (P = 0.002). These findings suggest an effect for RAD50 and NBS1 haploinsufficiency on genomic integrity and susceptibility to cancer. PMID- 16474177 TI - Ripe areca nut extract induces G1 phase arrests and senescence-associated phenotypes in normal human oral keratinocyte. AB - Around 200-600 million Asians chew areca (also called betel), which contains a mixture of areca nut and other ingredients. Epidemiological evidences indicated that areca use is tightly linked to oral carcinogenesis. This study investigated the effects of ripe areca nut extract (ANE) on cultured normal human oral keratinocyte (NHOK). Acute subtoxic ANE treatment inhibited DNA synthesis and induced cell cycle arrest at G1 phase in early passage (< 4th passage) cells. This was accompanied by a slight increase in the sub-G1 cellular fraction. O6 Methylguanine-DNA methyltransferase (MGMT), Hsp27 and p38MAPK was upregulated. p16 and p21 were remarkably upregulated early and declined afterwards. In contrast, the increase of dephosphorylated Rb seemed to be secondary to the episodes of p16 and p21 upregulation. To simulate the chronic areca exposure in vivo, constant ANE treatment in serial NHOK culture was performed. It resulted in a significant decrease in the population doubling, increase in senescence associated beta-galactosidase (SA-beta-Gal) and decrease in cell proliferation in NHOK of late passages (> or = 4th passage). Induction of senescence-associated phenotypes, G2/M accumulation and genomic instability following long-term ANE treatment were also observed in a low-grade oral carcinoma cell. ANE-treated NHOK also had a higher nuclear factor-kappaB (NF-kappaB) fraction and a lower cytosolic IkappaBalpha level relative to the control in late passages. Moreover, electrophoretic mobility shift assay (EMSA) indicated that ANE treatment shifted the NF-kappaB complex from high mobility position to lower mobility position in late-passaged NHOK. ANE treatment also upregulated IL-6 and cyclooxygenase-2 (COX 2) mRNA expressions in late-passaged NHOK. In summary, our findings suggest that ANE induces the cell cycle arrest at G1/S phase and the occurrence of senescence associated phenotypes of NHOK. The upregulation of p38MAPK, p16, p21, NF-kappaB, IL-6 and COX-2 are likely to participate in the control of these impacts. PMID- 16474178 TI - Loss of heterozygosity in human aberrant crypt foci (ACF), a putative precursor of colon cancer. AB - Aberrant crypt foci (ACF), the earliest neoplastic lesions of the colon, have genetic and epigenetic alterations. Loss of heterozygosity (LOH) of tumor suppressor gene loci is seen in most colon cancers, but it is not known how early in tumorigenesis this takes place. Nine microsatellite markers close to specific genes, that is, APC (5q21), PTPRJ (11p11), p53 (17p13) and DCC (18q21), were analyzed in 32 ACF and samples of normal crypts from the same 28 patients. Six losses of heterozygosity were found in 5 of 32 ACF: 4 losses of heterozygosity were at 11p11, the location of the gene for protein tyrosine phosphatase receptor type J (PTPRJ) and of a second independent region of deletion; the others were at 5q21 and 18q21. Microsatellite instability (MSI) with markers for a single locus was found in 4 of 32 ACF. All the observed allelic alterations (LOH and MSI) were in 8 of 32 ACF. The finding of LOH in ACF with normal expressions of adenomatous polyposis coli (APC) and beta-catenin proteins suggests that LOH can occur very early in colon neoplasia and perhaps even before APC mutations. The finding of 3 of 4 of the losses of heterozygosity at 11p11 for PTPRJ and half of all the losses of heterozygosity in this study at PTPRJ suggest that this gene plays a role early in colon neoplasia. PMID- 16474179 TI - The parity-related protection against breast cancer is compromised by cigarette smoke during rat pregnancy: observations on tumorigenesis and immunological defenses of the neonate. AB - Early pregnancy is a powerful negative risk factor for breast cancer (BCa) in women. Pregnancy also protects rats against induction of BCa by carcinogens such as N-methyl-N-nitrosourea (MNU), making the parous rat a useful model for studying this phenomenon. Smoking during early pregnancy may lead to an increased risk of BCa in later life, possibly attributable to carcinogens in cigarette smoke (CS), or to reversal of the parity-related protection against BCa. To investigate these possibilities, 50-day-old timed first-pregnancy rats were exposed to standardized mainstream CS (particle concentration = 50 mg/m3) or to filtered air (FA) 4 h/day, Day 2-20 of gestation. Age-matched virgin rats were similarly exposed to CS or FA. At age 100 days, the CS or FA-exposed, parous and virgin rats were injected s.c. with MNU (50 mg/kg body wt), or with MNU vehicle. Mammary tumors (MTs) first appeared in virgin rats 9 weeks post-MNU injection. While no MTs were detected in FA-exposed parous rats until 18 weeks post-MNU, MTs appeared in the CS-exposed parous rats as early as 10 wks (P < 0.02). As no MTs developed in CS-exposed rats not injected with MNU, CS did not act as a direct mammary carcinogen. Serum prolactin concentration on Day 19 of pregnancy in CS exposed dams was reduced by 50% compared with FA-exposed dams (P < 0.005). CS exposure during a pregnancy may thus 'deprotect' rats, enhancing their vulnerability to MNU-induced BCa. Prenatal CS exposure had no detectable effect on the immune responses of the pups examined at 3, 8 or 19 weeks of age. However, prolactin concentration in stomach contents (milk) of 3-day-old pups suckled by CS-exposed dams was decreased when compared with that of FA-exposed dams (P < 0.032). As milk-borne prolactin modulates development of the central nervous and immune systems of neonatal rats, CS exposure of the dams could adversely affect later maturation of these systems by reducing milk prolactin. PMID- 16474180 TI - Integrin-linked kinase regulates osteopontin-dependent MMP-2 and uPA expression to convey metastatic function in murine mammary epithelial cancer cells. AB - Metastasis-supporting physiological alterations are regulated by cell signaling molecules, which target signal transduction pathways and gene expression. Osteopontin (OPN) overexpression may represent a key molecular event in cancer metastasis. In this study, using metastatic 4T1 and non-metastatic 4T07 murine mammary cancer cell lines, we demonstrate that 4T1 cells exhibit significantly increased OPN, integrin-linked kinase (ILK), matrix metalloproteinase-2 (MMP-2) and urokinase-type plasminogen activator (uPA) expression in contrast to 4T07 cells. Blockade of OPN binding to 4T1 cell-surface integrins by the competitive ligand inhibitor, RGD, or a blocking antibody to alphavbeta3 integrin decreases OPN, ILK, MMP-2 and uPA expression. Conversely, exposure of 4T07 cells to exogenous OPN increases ILK, MMP-2 and uPA levels. Further experiments demonstrate that OPN-alphavbeta3 integrin binding in 4T1 with subsequent activation of ILK results in binding of AP-1 to MMP-2 and uPA promoter and increased in vitro promoter activation, as measured by transient transfection assays using MMP-2 and uPA promoter-reporter constructs. AP-1 activity is ablated by co-transfection of DN-ILK or exposure to RGD. Finally, functional correlative assays demonstrate that inhibition of ILK activity or RGD-mediated blockade of alphavbeta3 integrin binding significantly inhibits in vitro invasion, migration and invasion properties of 4T1 cells. In addition, uPA and MMP-2 have overlapping contributions to 4T1 migration and invasion characteristics. However, OPN and ILK activities contribute to 4T1 adhesion activities via mechanisms that are independent of uPA and MMP-2. Our results indicate that binding of an RGD-bearing ligand, such as OPN, to integrin receptors in metastatic 4T1 cells transcriptionally mediates MMP-2, uPA and OPN expression through ILK-dependent AP 1 activity and significantly increases in vitro functional correlates of metastasis. In 4T1 murine mammary cancer cells, we conclude that OPN mediates metastatic behavior, in part, through upregulation of MMP-2 and uPA protein expression. PMID- 16474181 TI - Resveratrol inhibits phorbol ester-induced expression of COX-2 and activation of NF-kappaB in mouse skin by blocking IkappaB kinase activity. AB - Aberrant expression of cyclooxygenase-2 (COX-2) has been implicated in tumor promotion. Resveratrol, a phytoalexin present in grapes, was reported to inhibit multistage mouse skin carcinogenesis. In the present study, we found that topically applied resveratrol significantly inhibited COX-2 expression induced by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA). Resveratrol suppressed phosphorylation and subsequent degradation of IkappaBalpha, thereby inhibiting activation of nuclear factor-kappaB (NF-kappaB) in TPA-stimulated mouse skin. Pretreatment with resveratrol also suppressed TPA-induced phosphorylation of extracellular signal-regulated protein kinase (ERK) and p38 mitogen-activated protein (MAP) kinase. Resveratrol blunted TPA-induced phosphorylation of p65 and its interaction with CBP/p300, rendering NF-kappaB transcriptionally inactive. To get further insights into the molecular basis of NF-kappaB inactivation by resveratrol, we examined the role of IkappaB kinase (IKK) in mediating TPA-induced activation of NF-kappaB and COX-2 expression. TPA treatment led to rapid induction of IKK activity in mouse skin, which was abolished either by resveratrol or an IKK inhibitor Bay 11-7082. Topical application of Bay 11-7082 also abrogated TPA-induced NF-kappaB activation and COX-2 expression, supporting the involvement of IKK in TPA-induced COX-2 expression. Taken together, the above findings suggest that resveratrol targets IKK in blocking TPA-induced NF-kappaB activation and COX-2 expression in mouse skin in vivo. PMID- 16474183 TI - Relevance of ventricular electrical dispersion to arrhythmogenesis in ischemic myocardium--a simulation study. AB - A computer simulation method was used to study the possible role of electrical dispersion induced by regional ischemia in the mechanisms underlying cardiac arrhythmias. Ischemic cells were simulated by considering the three major component conditions of acute ischemia (elevated extracellular K+ concentration, acidosis and anoxia) at the level of ionic currents and ionic concentrations. An ischemic area was introduced into a homogeneous healthy tissue to create a localized inhomogeneity. The constructed models were solved using the operator splitting and adaptive time step methods. The numerical experiments showed that action potential durations (APDs) of ischemic cells did not change with beats of shorter or longer cycle length. The smaller percentage increase of slow component of the delayed rectifier K+ current, I(ks), and smaller outward Na+-Ca2+ exchange current were found to be the ionic mechanisms underlying the decreased rate dependence in ischemic cells. The results suggest that ischemia flattens the APD restitution curve; however, the dispersion of refractory period can be greatly increased by a premature beat in the constructed inhomogeneous sheet. This demonstrates that the dispersion of refractoriness rather than APD by a premature beat contributes to reentrant tachyarrhythmias in the locally ischemic tissue. PMID- 16474184 TI - Slow and fast fatigable frog muscle fibres: electrophysiological and histochemical characteristics. AB - Continuous activity of isolated frog gastrocnemius muscle fibres provoked by repetitive stimulation of 5 Hz was used as an experimental model for fatigue development in different fibre types. Parameter changes of the elicited intracellular action potentials and mechanical twitches during the period of uninterrupted activity were used as criteria for fatigue evaluation. Slow fatigable muscle fibre (SMF) and fast fatigable muscle fibre (FMF) types were distinguished depending on the duration of their uninterrupted activity, which was significantly longer in SMFs than in FMFs. The normalized changes of action potential amplitude and duration were significantly smaller in FMFs than in SMFs. The average twitch force and velocity of contraction and relaxation were significantly higher in FMFs than in SMFs. Myosin ATPase (mATPase) and succinate dehydrogenase activity were studied by histochemical assessment in order to validate the fibre type classification based on their electrophysiological characteristics. Based on the relative mATPase reactivity, the fibres of the studied muscle were classified as one of five different types (1-2, 2, 2-3, 3 and tonic). Smaller sized fibres (tonic and type 3) expressed higher succinate dehydrogenase activity than larger sized fibres (type 1-2, 2), which is related to the fatigue resistance. The differences between fatigue development in SMFs and FMFs during continuous activity were associated with fibre-type specific mATPase and succinate dehydrogenase activity. PMID- 16474185 TI - The ACAT inhibitor VULM1457 significantly reduced production and secretion of adrenomedullin (AM) and down-regulated AM receptors on human hepatoblastic cells. AB - Acyl-CoA:cholesterol acyltransferase (ACAT) is an important enzyme in the pathways of cholesterol esterification. It has been shown that new ACAT inhibitor 1-(2,6-diisopropyl-phenyl)-3-[4-(4'-nitrophenylthio)phenyl] urea (VULM1457) significantly reduced atherogenic activity in animal experimental atherosclerosis. Proliferative hormone adrenomedullin (AM) has been shown to be released in response to hypoxia, however, its role in cellular protection has remained elusive. The effect of increased local production of AM in cells and resultant down-regulation of AM receptors has not been investigated yet. We hypothesized that increased expression of AM in hypoxic cells was the result of excessive AM production with resultant AM receptor down-regulation, surface membrane protein degradation and that the new specific ACAT inhibitor would reduce AM induction in hypoxia and thus proliferation of cells. In order to investigate specific cellular AM signaling and protection induced by VULM1457, we characterized specific surface-membrane [125I]AM receptors expressed on cells, evaluated AM secretion (RIA assays), AM mRNA expression in cultured cells (RT-PCR analysis) and proliferation (incorporation of [3H]thymidine) in control, hypoxic and metabolically stressed human hepatoblastoma cell lines exposed to gradually increasing concentrations of VULM1457. The new ACAT inhibitor VULM1457 in concentration 0.03 and 0.1 micromol/l significantly down-regulated specific AM receptors on HepG2 cells, reduced AM secretion of HepG2 cells exposed to hypoxia. These results suggest that VULM1457, as new member of ACAT family of inhibitors could negatively regulate cell proliferation induced by AM, which may correlate with down-regulation of membrane-bound AM receptors on HepG2 cells, and moreover, with the induction and expression of AM in hypoxia. PMID- 16474187 TI - Tapering of human nerve fibres. AB - To determine the tapering of human nerve fibres, rostral and caudal root pieces of cauda equina nerve roots were removed and nerve fibre diameter distributions were constructed for 4 myelin sheath thickness ranges for the two sites, and compared with each other. The reduction of the group diameter in the different alpha-motoneuron groups was 0.2 % per 13 cm. Accounting for systematic errors, there may be even less tapering. An identified single nerve fibre showed no tapering. Further, there is indication that gamma-motoneurons, preganglionic sympathetic and parasympathetic fibres and skin afferents also reduce their fibre diameter by 0.2 % per 13 cm or less. Consequently, a nerve fibre with a diameter of 10 microm would be reduced to approximately 9.8 microm at 1m from the cell soma. Preganglionic parasympathetic fibres were found to be represented in roots S1 to S5. At similar distances from the spinal cord, the mean diameter of ventral root alpha1-motoneuron (FF) axons increased from the thoracic towards the lumbo sacral region before decreasing again in the lower sacral region. Usually no alpha1-motoneuron axons were found in S5 roots. The diameter distribution of unmyelinated nerve fibres of a ventral S5 root showed three peaks at 0.25, 0.95 and 1.2 microm. The unmyelinated fibres with diameters around 0.25 microm may represent parasympathetic fibres. In six selected areas of the ventral S5 root, 6.6 times more unmyelinated nerve fibres than myelinated fibres were found on the average. PMID- 16474186 TI - The role of NO in ischemia/reperfusion injury in isolated rat heart. AB - Nitric oxide (NO) is an important regulator of myocardial function and vascular tone under physiological conditions. However, its role in the pathological situations, such as myocardial ischemia is not unequivocal, and both positive and negative effects have been demonstrated in different experimental settings including human pathology. The aim of the study was to investigate the role of NO in the rat hearts adapted and non-adapted to ischemia. Isolated Langendorff perfused hearts were subjected to test ischemic (TI) challenge induced by 25 min global ischemia followed by 35 min reperfusion. Short-term adaptation to ischemia (ischemic preconditioning, IP) was evoked by 2 cycles of 5 min ischemia and 5 min reperfusion, before TI. Recovery of function at the end of reperfusion and reperfusion-induced arrhythmias served as the end-points of injury. Coronary flow (CF), left ventricular developed pressure (LVDP), and dP/dt(max) (index of contraction) were measured at the end of stabilization and throughout the remainder of the protocol until the end of reperfusion. The role of NO was investigated by subjecting the hearts to 15 min perfusion with NO synthase (NOS) inhibitor L-NAME (100 mmol/l), prior to sustained ischemia. At the end of reperfusion, LVDP in the controls recovered to 29.0 +/- 3.9 % of baseline value, whereas preconditioned hearts showed a significantly increased recovery (LVDP 66.4 +/- 5.7 %, p < 0.05). Recovery of both CF and dP/dt(max) after TI was also significantly higher in the adapted hearts (101.5 +/- 5.8 % and 83.64 +/- 3.92 % ) as compared with the controls (71.9 +/- 6.3 % and 35.7 +/- 4.87 %, respectively, p < 0.05). NOS inhibition improved contractile recovery in the non adapted group (LVDP 53.8 +/- 3.1 %; dP/dt(max) 67.5 +/- 5.92 %) and increased CF to 82.4 +/- 5.2 %. In contrast, in the adapted group, it abolished the protective effect of IP (LVDP 31.8 +/- 3.1 %; CF 70.3 +/- 3.4 % and dP/dt(max) 43.25 +/- 2.19 %). Control group exhibited 100 % occurrence of ventricular tachycardia (VT), 57 % incidence of ventricular fibrillation (VF) - 21 % of them was sustained VF (SVF); application of L-NAME attenuated reperfusion arrhythmias (VT 70 %, VF 20 %, SVF 0 %). Adaptation by IP also reduced arrhythmias, however, L NAME in the preconditioned hearts increased the incidence of arrhythmias (VT 100 %, VF 58 %, SVF 17 %). IN CONCLUSION: our results indicate that administration of L-NAME might be cardioprotective in the normal hearts exposed to ischemia/reperfusion (I/R) alone, suggesting that NO contributes to low ischemic tolerance in the non-adapted hearts. On the other hand, blockade of cardioprotective effect of IP by L-NAME points out to a dual role of NO in the heart: a negative role in the non-adapted myocardium subjected to I/R, and a positive one, due to its involvement in the mechanisms of protection triggered by short-term cardiac adaptation by preconditioning. PMID- 16474188 TI - ESR and monolayer study of the localization of coenzyme Q10 in artificial membranes. AB - The data obtained from the ESR experiments show a complex, depth dependent effect of CoQ10 on the lipid molecules mobility in the bilayer. These effects depend both on its concentration and the temperature. CoQ10 disturbs not only the hydrophobic core of the membrane but also the region close to the hydrophilic headgroups of phospholipids. Both these effects could be explained by the fact that the high hydrophobicity of CoQ10 causes the molecules to position itself in the interior of the bilayer, but at the same time its water seeking headgroup is located close to the region of the polar headgrops of membrane lipids. The presence of CoQ10 in the hydrophobic core has further implications on the properties of membrane intrinsic domain. Results of monolayer experiments indicate that CoQ10 may form aggregates when mixed with PC molecules in the lipid hydrocarbon chain-length dependent manner. CoQ10 is not fully miscible with DMPC or DPPC but it is well miscible with the long-chain DSPC molecules. Our suggestion is that CoQ10 when present in long-chain phospholipid bilayer, interacts with saturated fatty acyl-chains and adapt the structure which allows such interactions: either parallel to the saturated acyl chains or "pseudo-ring" conformation resembling sterol structure. PMID- 16474190 TI - Energy and glucose pathways in thiamine deficient primary rat brain microvascular endothelial cells. AB - Thiamine deficiency (TD) results in lactate acidosis, which is associated with neurodegeneration. The aim of this study was to investigate this alteration in primary rat brain endothelia. Spectrophotometric analysis of culture media revealed that only a higher concentration of pyrithiamine, which accelerates the intracellular blocking of thiamine, significantly elevated the lactate level and lactate dehydrogenase activity within 7 days. The medium without pyrithiamine and with a thiamine concentration comparable to pathophysiological plasma levels mildly reduced only the activity of transketolase. This suggests that significant metabolic changes may not occur at the early phase of TD in cerebral capillary cells, while anaerobic glycolysis in capillaries may be mediated during late stage/chronic TD. PMID- 16474189 TI - Prolongation of pentoxifylline aliphatic side chain positively affects the reversal of P-glycoprotein-mediated multidrug resistance in L1210/VCR line cells. AB - We reported previously that derivatives of pentoxifylline (PTX) reverse multidrug resistance (MDR) in P-glycoprotein (P-gp) positive L1210/VCR cells. Based on the results of a recent study using 25 N-alkylated methylxanthines with carbohydrate side-chains of various lengths, we formulated the following design criteria for a methylxanthine molecule to effectively reverse P-gp mediated MDR: i) a massive substituent at the N1 position is crucial for MDR reversal potency; ii) elongation of the substituents at the N3 and N7 positions (from methyl to propyl) increases the efficacy of a xanthine to reverse MDR; iii) elongation of the substituent at the C8 position (from H to propyl) decreases the efficacy of a xanthine to reverse MDR. Based on these criteria, we synthesized and tested for potency to reverse MDR a new PTX derivative, 1-(10-undecylenyl)-3-heptyl-7-methyl xanthine (PTX-UHM), with prolonged substituents at the N1 and N3 positions. The derivative was obtained by alkylation of 3-heptyl-7-methyl xanthine with 1 methylsulfonyloxy-10-undecylenyl. NMR and IR structural analyses proved the identity of the product. Cytotoxicity study showed that PTX-UHM is only slightly more toxic to L1210/VCR cells than PTX. We found that both PTX-UHM and PTX were able to reverse vincristine resistance of L1210/VCR cells, yet PTX-UHM was significantly more efficient in the reversal than PTX. PMID- 16474191 TI - Primary cilia of inv/inv mouse renal epithelial cells sense physiological fluid flow: bending of primary cilia and Ca2+ influx. AB - Primary cilia are hypothesized to act as a mechanical sensor to detect renal tubular fluid flow. Anomalous structure of primary cilia and/or impairment of increases in intracellular Ca2+ concentration in response to fluid flow are thought to result in renal cyst formation in conditional kif3a knockout, Tg737 and pkd1/pkd2 mutant mice. The mutant inv/inv mouse develops multiple renal cysts like kif3a, Tg737 and pkd1/pkd2 mutants. Inv proteins have been shown to be localized in the renal primary cilia, but response of inv/inv cilia to fluid stress has not been examined. In the present study, we examined the mechanical response of primary cilia to physiological fluid flow using a video microscope, as well as intracellular Ca2+ increases in renal epithelial cells from normal and inv/inv mice in response to flow stress. Percentages of ciliated cells and the length of primary cilia were not significantly different between primary renal cell cultures from normal and inv/inv mutant mice. Localization of inv protein was restricted to the base of primary cilia even under flow stress. Inv/inv mutant cells had similar bending mechanics of primary cilia in response to physiological fluid flow compared to normal cells. Furthermore, no difference was found in intracellular Ca2+ increases in response to physiological fluid flow between normal and inv/inv mutant cells. Our present study suggests that the function of the inv protein is distinct from polaris (the Tg737 gene product), polycystins (pkd1 and pkd2 gene products). PMID- 16474192 TI - Purification and characterization of a thermostable-cellulase free xylanase from Syncephalastrum racemosum Cohn. AB - Syncephalastrum racemosum Cohn. produces an extracellular xylanase that was shown to potentially bleach pulp at pH 10 and 50 degrees C. The enzyme was found to be a dimer with an apparent molecular weight of 29 kDa as determined by SDS-PAGE. The optimum activity was found at two pH values 8.5 and 10.5; however the activity sharply decreased below pH 6 and above pH 10.5. The enzyme was stable for 72 h at pH 10.5 and at 50 degrees C. Kinetic experiments at 50 degrees C gave V(max) and K(m) of 1,400 U/ml min(-1) mg(-1) protein and 0.05 mg/ml respectively. The enzyme had no apparent requirement for cofactors, and its activity was strongly inhibited by group II b metal ions like Zn2+, Hg2+, etc. Xylan completely protected the enzyme from being inactivated by N-bromosuccinimide. PMID- 16474193 TI - Bullera hoabinhensis sp. nov., a new ballistoconidiogenous yeast isolated from a plant leaf collected in Vietnam. AB - VY-68, a ballistoconidiogenous yeast strain, isolated from a plant leaf at Cuc Phuong National Park of Ninh Binh Province, Vietnam, was assigned to the genus Bullera based on morphological and chemotaxonomical characteristics. Based on the sequence analyses of 18S rDNA, D1/D2 region of 26S rDNA, and internal transcribed spacer regions (ITS), VY-68 was phylogenetically closely related to Bullera pseudoalba and Cryptococcus cellulolyticus. DNA-DNA reassociation experiments among VY-68, B. pseudoalba and C. cellulolyticus revealed that strain VY-68 is a distinct species, and the latter two are conspecific. Bullera hoabinhensis is proposed for VY-68. PMID- 16474194 TI - Sequence analysis of partial toxR gene from Philippine Vibrio isolates and design of toxR-targeted primers for detection. AB - Vibriosis in penaeid species cultured in the Philippines results in massive mortalities and consequently in severe economic losses in the shrimp industry. Rapid and accurate detection of the causative agent of the disease is imperative. In this study, toxR gene sequence analysis of ten Vibrio isolates (from several provinces of the Philippines) implicated in disease affecting the penaeid shrimp (Penaeus monodon) was performed in order to develop a toxR-targeted PCR detection of similar strains of shrimp pathogens. Analysis of the partial toxR gene revealed 97-100% sequence similarity among the ten Philippine Vibrio isolates. Distinct sequence variation of the toxR gene, however, was observed between the Philippine Vibrio isolates and the type strains, with the Philippine isolates exhibiting only 92-93% and 74-75% sequence similarity with the type strain V. campbellii (NBRC 15631T) and V. harveyi (NBRC 15634T), respectively. The use of a PCR primer set that was designed based on toxR sequences of the Philippine Vibrio isolates amplified the expected 226-bp toxR fragment using templates from all ten Philippine Vibrio isolates. No amplified product was observed in PCR using templates from type strains of V. harveyi, V. campbellii, and other non-target bacteria, suggesting that the primers were specific for the Philippine Vibrio isolates. The toxR-targeted PCR primers reported in this study could be useful in the detection of Philippine Vibrio isolates associated with mortalities in the shrimp industry, which could not be detected in PCR using primers designed for type strains of V. harveyi and V. campbellii. PMID- 16474195 TI - Isolation and characterization of mesophilic cellulose-degrading bacteria from flower stalks-vegetable waste co-composting system. AB - Fifteen mesophilic bacteria with high C(x) cellulase activities were isolated and purified from a mixed-culture enriched from a flower stalks-vegetable waste co composting system. A CMCase test showed that the enzyme activity of these isolates ranged from 7.9 to 28.0 U ml(-1). Although filter paper degrading capability was low in single culture, significant synergetic cellulose degradation were detected in four groups of mixed cultures, their degradation rates were 23.5%, 26.3%, 19.4% and 24.5%, respectively. Study of morphological and physiological characters of five predominant isolates which possess high CMCase and had positive effect on synergetic cellulose degradation in mixed culture system showed that two of them were closely related to Bacillus pasteurii and Bacillus cereus, whereas the rest belong to the genus Halobacillus, Aeromicrobium and Brevibacterium, respectively. PMID- 16474196 TI - Distribution and persistence of fecal bacterial populations in liquid and dewatered sludge from a biological treatment plant. AB - The changes in composition and structure of fecal coliforms (FC) and enterococci (ENT) populations, as well as the elimination of spores of sulphite-reducing bacteria (SRB), were compared between municipal sewage and their derived sludge in a biological treatment plant in order to determine any selective reduction or adsorption to sludge during the treatment process. Additionally, the persistence of antibiotic-resistant enterococcal populations in two kinds of sludge was also considered to evaluate their potential elimination in the treatment process. Microbial indicators, vancomycin-resistant and erythromycin-resistant enterococci were enumerated. The structure and composition of FC and ENT populations were determined by biochemical fingerprinting and clustering analyses. Raw and treated sewage showed a concentration of FC 1 log unit higher than ENT and nearly 2 log units higher than spores of SRB. However, the three studied indicators showed similar concentrations in both types of sludge. Consequently, FC were eliminated in higher proportion than ENT and spores of SRB in sludge. FC and ENT populations showed high diversity and similarity population indexes for all kinds of samples. Antibiotic-resistant enterococci persisted in a similar proportion in respect to total enterococci not only in treated sewage but also in sludge. The persistence of antibiotic-resistant strains in sludge as well as in treated sewage should be considered if they are used for land disposal or for water reutilization, respectively. PMID- 16474197 TI - Genomic diversity of Astragalus cicer microsymbionts revealed by AFLP fingerprinting. AB - DNA polymorphism among 36 Astragalus cicer nodule isolates and 9 reference mesorhizobia was evaluated by a simplified PstI based AFLP procedure with three selective primers: Pst-A, Pst-G, and Pst-GC. The DNA profiles were found to be highly specific for nearly each strain, although DNA bands characteristic for most A. cicer microsymbionts were also noted. The overall topologies of dendrograms, generated by AFLP patterns in PCR reaction with three primers, were very similar to one another and to that constructed by phenotyping. Also the strain compositions in the particular clusters on pheno- and genomograms were in good agreement. The obtained results indicate that AFLP technique can be a useful tool for typing of A. cicer rhizobia as well as for studying their diversity. PMID- 16474198 TI - Effects of air-drying soils on discriminating the soils with community-level physiological profiling. PMID- 16474199 TI - Phylogenetic analysis based on 16S rRNA gene sequences of deep-sea bacteria isolated from intestinal contents of deep-sea fishes retrieved from the abyssal zone. PMID- 16474200 TI - Study of the heterogeneity of 16S rRNA genes in gamma-proteobacteria: implications for phylogenetic analysis. PMID- 16474201 TI - Mechanisms of the antinociceptive action of gabapentin. AB - Gabapentin, a gamma-aminobutyric acid (GABA) analogue anticonvulsant, is also an effective analgesic agent in neuropathic and inflammatory, but not acute, pain systemically and intrathecally. Other clinical indications such as anxiety, bipolar disorder, and hot flashes have also been proposed. Since gabapentin was developed, several hypotheses had been proposed for its action mechanisms. They include selectively activating the heterodimeric GABA(B) receptors consisting of GABA(B1a) and GABA(B2) subunits, selectively enhancing the NMDA current at GABAergic interneurons, or blocking AMPA-receptor-mediated transmission in the spinal cord, binding to the L-alpha-amino acid transporter, activating ATP sensitive K(+) channels, activating hyperpolarization-activated cation channels, and modulating Ca(2+) current by selectively binding to the specific binding site of [(3)H]gabapentin, the alpha(2)delta subunit of voltage-dependent Ca(2+) channels. Different mechanisms might be involved in different therapeutic actions of gabapentin. In this review, we summarized the recent progress in the findings proposed for the antinociceptive action mechanisms of gabapentin and suggest that the alpha(2)delta subunit of spinal N-type Ca(2+) channels is very likely the analgesic action target of gabapentin. PMID- 16474202 TI - Induction of heme oxygenase-1 inhibits monocyte chemoattractant protein-1 mRNA expression in U937 cells. AB - Heme oxygenase-1 (HO-1) is a stress-inducible isoform of HO with potential cytoprotective effects. Monocyte activation/migration mediated by monocyte chemoattractant protein-1 (MCP-1) is one of the earliest and important events in the pathogenesis of atherosclerosis. We examined the effect of HO-1 on the production of lysophosphatidylcholine (Lyso-PC)-induced MCP-1 in the human promonocytic cell line U937. Increased HO-1 induction by hemin resulted in a significant decrease in the Lyso-PC-mediated induction of MCP-1 mRNA expression. SnPP (IX), the specific inhibitor of HO-1 enzymatic activity, prevented the hemin mediated attenuation of MCP-1 mRNA expression. These results suggest that HO-1 may work as an anti-atherogenic agent through the attenuation of MCP-1 production. PMID- 16474203 TI - Generalization of NMDA-receptor antagonists to the discriminative stimulus effects of kappa-opioid receptor agonists U-50,488H, but not TRK-820 in rats. AB - Generalizations of NMDA-receptor antagonists to the discriminative stimulus effects of kappa-opioid receptor agonists in rats were examined. Phencyclidine, MK-801, and ketamine, non-competitive NMDA-receptor antagonists, generalized to the discriminative stimulus effects of U-50,488H, but not those of TRK-820, whereas (+/-)-3-(2-carbaxypiperazine-4-yl) propyl-1-phosphonic acid (CPP), a competitive NMDA-receptor antagonist, and ifenprodil, an NR1/NR2B NMDA-receptor antagonist, did not, suggesting that non-competitive NMDA-receptor antagonists possess U-50,488H-like discriminative stimulus effects in rats. Since U-50,488H and phencyclidine both induce aversive effects, our findings indicate that the cue of the discriminative stimulus effects of U-50,488H and non-competitive NMDA receptor antagonists may be associated with their aversive effects. PMID- 16474204 TI - Benidipine, a calcium channel blocker, regulates proliferation and phenotype of vascular smooth muscle cells. AB - Hyperproliferation of phenotypically modified vascular smooth muscle cells (VSMCs) is one of the major factors in the development of atherosclerosis and restenosis. Previously it was demonstrated that benidipine, a dihydropyridine calcium channel antagonist, reduced neointimal formation in a rat balloon-injury model. In the present study, we examined the effect of benidipine on the phenotypic modulation and proliferation of VSMCs, using primary cultures of rat VSMCs. In the absence of drug treatment, protein levels of the smooth muscle specific markers, such as smooth muscle myosin heavy chain-1 (SM1), calponin 1, and alpha-actin, decreased during culture. However, treatment of VSMCs with benidipine (3 - 10 micromol/L) for 1 week reversed the effect in a concentration related manner so that high levels of marker proteins were maintained. The expression of calponin mRNAs was reduced markedly during 1-week culture, and treatment with benidipine (3 micromol/L) significantly inhibited the reduction. Treatment with benidipine for 2 days increased the level of p21 protein and partially reduced p70 S6 kinase 1 (p70S6K1) activity. These data suggest that benidipine may arrest the growth of VSMCs, thereby preventing cell dedifferentiation. These additional properties of benidipine suggest that the drug should provide useful therapy for atherosclerosis and restenosis. PMID- 16474205 TI - Protective effect of zinc against ischemic neuronal injury in a middle cerebral artery occlusion model. AB - In this study, we investigated the effect of vesicular zinc on ischemic neuronal injury. In cultured neurons, addition of a low concentration (under 100 microM) of zinc inhibited both glutamate-induced calcium influx and neuronal death. In contrast, a higher concentration (over 150 microM) of zinc decreased neuronal viability, although calcium influx was inhibited. These results indicate that zinc exhibits biphasic effects depending on its concentration. Furthermore, in cultured neurons, co-addition of glutamate and CaEDTA, which binds extra-cellular zinc, increased glutamate-induced calcium influx and aggravated the neurotoxicity of glutamate. In a rat transient middle cerebral artery occlusion (MCAO) model, the infarction volume, which is related to the neurotoxicity of glutamate, increased rapidly on the intracerebral ventricular injection of CaEDTA 30 min prior to occlusion. These results suggest that zinc released from synaptic vesicles may provide a protective effect against ischemic neuronal injury. PMID- 16474207 TI - A novel effect of bifemelane, a nootropic drug, on intracellular Ca2+ levels in rat cerebral astrocytes. AB - We investigated the effects of bifemelane, a nootropic drug, on the intracellular calcium concentration ([Ca2+]i) in rat cerebral astrocytes using a Ca2+ imaging device. At concentrations of 10 - 30 microM, bifemelane induced a slow onset and small increase in the [Ca2+]i, while at higher concentrations (100 - 300 microM), it induced a rapid transient increase in the [Ca2+]i during administration and a second large increase was seen during drug washout. The first peak was observed in Ca2+-free medium, but its onset was significantly delayed, and no second peak was seen. Neither of these effects was seen in cells treated with thapsigargin, a specific inhibitor of endoplasmic reticulum Ca2+-ATPase, in Ca2+-free medium. When thapsigargin-treated astrocytes were returned to normal medium containing Ca2+ (1.8 mM), the [Ca2+]i increased significantly, and this effect was reversely inhibited by bifemelane. We conclude that bifemelane causes the first peak by stimulating release from intracellular Ca2+ stores and the second by capacitive entry through store-operated Ca2+ channels. Although the detail mechanisms of action of the drug are still unknown, bifemelane will be provided as a pharmacological tool for basic studies on astrocytes. PMID- 16474206 TI - Inhibitory mechanism of monensin on high K+-induced contraction in guniea-pig urinary bladder. AB - In this study, we examined the inhibitory mechanism of monensin on high K+ induced contraction in guinea-pig urinary bladder. The relaxant effect of monensin (0.001 - 10 microM) was more potent than those of NaCN (100 microM - 1 mM) and forskolin (3 - 10 microM). Monensin (0.1 microM), NaCN (300 microM), or forskolin (10 microM) inhibited high K+-induced contraction without decreasing [Ca2+]i level. Monensin and NaCN remarkably decreased creatine phosphate and ATP contents. Monensin and NaCN inhibited high K+-induced increases in flavoprotein fluorescence, which is involved in mitochondrial respiration. Forskolin increased cAMP content but monensin did not. Monensin increased Na+ content at 10 microM but not at 0.1 microM that induced maximum relaxation. In the alpha-toxin permeabilized muscle, forskolin significantly inhibited the Ca2+-induced contraction, but monensin did not affect it. These results suggest that the relaxation mechanism of monensin in smooth muscle of urinary bladder may be an inhibition of oxidative metabolism. PMID- 16474208 TI - c-Jun N-terminal kinase contributes to norepinephrine-induced contraction through phosphorylation of caldesmon in rat aortic smooth muscle. AB - Vascular smooth muscle contraction is mediated by activation of extracellular signal-regulated kinase (ERK) 1/2, an isoform of mitogen-activated protein kinase (MAPK). However, the role of stress-activated protein kinase/c-Jun N-terminal kinase (JNK) in vascular smooth muscle contraction has not been defined. We investigated the role of JNK in the contractile response to norepinephrine (NE) in rat aortic smooth muscle. NE evoked contraction in a dose-dependent manner, and this effect was inhibited by the JNK inhibitor SP600125. NE increased the phosphorylation of JNK, which was greater in aortic smooth muscle from hypertensive rats than from normotensive rats. NE-induced JNK phosphorylation was significantly inhibited by SP600125 and the conventional-type PKC (cPKC) inhibitor Go6976, but not by the Rho kinase inhibitor Y27632 or the phosphatidylinositol 3-kinase inhibitor LY294002. Thymeleatoxin, a selective activator of cPKC, increased JNK phosphorylation, which was inhibited by Go6976. SP600125 attenuated the phosphorylation of caldesmon, an actin-binding protein whose phosphorylation is increased by NE. These results show that JNK contributes to NE-mediated contraction through phosphorylation of caldesmon in rat aortic smooth muscle, and that this effect is regulated by the PKC pathway, especially cPKC. PMID- 16474209 TI - The sigma1 protein as a target for the non-genomic effects of neuro(active)steroids: molecular, physiological, and behavioral aspects. AB - Steroids synthesized in the periphery or de novo in the brain, so called 'neurosteroids', exert both genomic and nongenomic actions on neurotransmission systems. Through rapid modulatory effects on neurotransmitter receptors, they influence inhibitory and excitatory neurotransmission. In particular, progesterone derivatives like 3alpha-hydroxy-5alpha-pregnan-20-one (allopregnanolone) are positive allosteric modulators of the gamma-aminobutyric acid type A (GABA(A)) receptor and therefore act as inhibitory steroids, while pregnenolone sulphate (PREGS) and dehydroepiandrosterone sulphate (DHEAS) are negative modulators of the GABA(A) receptor and positive modulators of the N methyl-D-aspartate (NMDA) receptor, therefore acting as excitatory neurosteroids. Some steroids also interact with atypical proteins, the sigma (sigma) receptors. Recent studies particularly demonstrated that the sigma1 receptor contributes effectively to their pharmacological actions. The present article will review the data demonstrating that the sigma1 receptor binds neurosteroids in physiological conditions. The physiological relevance of this interaction will be analyzed and the impact on physiopathological outcomes in memory and drug addiction will be illustrated. We will particularly highlight, first, the importance of the sigma1 receptor activation by PREGS and DHEAS which may contribute to their modulatory effect on calcium homeostasis and, second, the importance of the steroid tonus in the pharmacological development of selective sigma1 drugs. PMID- 16474210 TI - PKA implicated in the phosphorylation of Cx43 induced by stimulation with FSH in rat granulosa cells. AB - Connexin 43 (Cx43)-mediated gap junctional communication in granulosa cells is crucial for germ line development and postnatal folliculogenesis. We previously showed that follicle-stimulating hormone (FSH) promoted phosphorylation of Cx43 in rat primary granulosa cells. We further identified Ser365, Ser368, Ser369, and Ser373 in the carboxy-terminal tail as the major sites of phosphorylation by FSH, and found that the phosphorylation of these residues was essential for channel activity. In this study, we investigated the protein kinase(s) responsible for FSH-induced phosphorylation. H89, a cyclic AMP-dependent protein kinase (PKA) inhibitor, inhibited FSH-induced phosphorylation both in vivo and in vitro, whereas PD98059, a mitogen-activated protein kinase kinase (MEK) inhibitor, had little effect on the phosphorylation level. Ca2+-dependent protein kinase (PKC) appeared to negatively regulate phosphorylation. Phosphopeptide mapping with or without H89 treatment indicated that PKA could be responsible for phosphorylation of the four serine residues. In addition, the purified catalytic subunit of PKA could phosphorylate the recombinant C-terminal region of Cx43, but not the variant in which all four serine residues were substituted with alanine. These results suggest that FSH positively regulates Cx43-mediated channel formation and activity through phosphorylation of specific sites by PKA. PMID- 16474211 TI - Epigenetic characterization of the CpG islands of bovine Leptin and POU5F1 genes in cloned bovine fetuses. AB - Abnormal development and fetal loss during postimplantation period are concerns for production of nuclear transferred animals. Aberrant DNA methylation is one of the reasons for poor survival of cloned animals. In mammalian genome DNA, CpG islands are preferentially located at the start of transcription of housekeeping genes and are associated with tissue-specific genes. The correct and consecutive mechanisms of DNA methylation in the CpG islands are necessary for selective gene expressions that determine the properties of individual cells, tissues, and organs. In this study, we investigated the methylation status of the CpG islands of the bovine Leptin and POU5F1 genes in fetal and placental tissues from fetuses produced by artificial insemination (AI) and nuclear transfer (NT) at days 48 and 59 of pregnancy. Altered DNA methylation was observed in the normal and cloned fetal, placental, and endometrial tissues using bisulfite sequencing and pyrosequencing. Different tissue-specific methylated regions in the bovine Leptin and POU5F1 genes show a variable methylation status in NT fetuses compared to AI control. PMID- 16474213 TI - Role of transcranial approaches in the treatment of sellar and suprasellar lesions. AB - The principles in the surgical management of sellar and suprasellar tumors are to relieve mass effect, normalize pituitary hypersecretion, preserve or restore normal pituitary function, prevent tumor recurrence and to provide tissue for pathological and scientific study. Over the past century, the transsphenoidal approach has evolved as the approach of choice for pituitary surgeons. Despite the limitations of transcranial approaches in accessing the intrasellar component of pituitary adenomas and historically their increased morbidity and mortality, there are situations where transcranial procedures have considerable advantages over transsphenoidal approaches. As a consequence, transcranial approaches retain an essential role in the treatment of certain sellar and suprasellar tumors and it remains necessary for all pituitary surgeons to master this approach. PMID- 16474212 TI - DNA methylation profiles of donor nuclei cells and tissues of cloned bovine fetuses. AB - Methylation of DNA in CpG islands plays an important role during fetal development and differentiation because CpG islands are preferentially located in upstream regions of mammalian genomic DNA, including the transcription start site of housekeeping genes and are also associated with tissue-specific genes. Somatic nuclear transfer (NT) technology has been used to generate live clones in numerous mammalian species, but only a low percentage of nuclear transferred animals develop to term. Abnormal epigenetic changes in the CpG islands of donor nuclei after nuclear transfer could contribute to a high rate of abortion during early gestation and increase perinatal death. These changes have yet to be explored. Thus, we investigated the genome-wide DNA methylation profiles of CpG islands in nuclei donor cells and NT animals. Using Restriction Landmark Genomic Scanning (RLGS), we showed, for the first time, the epigenetic profile formation of tissues from NT bovine fetuses produced from cumulus cells. From approximately 2600 unmethylated NotI sites visualized on the RLGS profile, at least 35 NotI sites showed different methylation statuses. Moreover, we proved that fetal and placental tissues from artificially inseminated and cloned cattle have tissue specific differences in the genome-wide methylation profiles of the CpG islands. We also found that possible abnormalities occurred in the fetal brain and placental tissues of cloned animals. PMID- 16474214 TI - Extended transsphenoidal approach. AB - Transsphenoidal surgery is well established as an effective primary treatment for tumors of the sellar region. The technique of transsphenoidal surgery has evolved over the years with many prominent surgeons contributing to its present state of refinement. The transsphenoidal approach can be modified in various ways to permit resection of parasellar tumors that otherwise would require a transcranial or transbasal operation. Our experience with these 'extended' techniques has primarily involved the transtuberculum sella approach in which bone is removed from the tuberculum sellae and the posterior portion of the planum sphenoidale. Experience with this technique continues to burgeon, and offers an excellent alternative to transcranial surgery in dealing with a difficult constellation of tumors. Meticulous attention to detail, particularly with respect to reconstruction and closure of the sellar floor, is necessary for its effective application. PMID- 16474215 TI - Image guidance in pituitary surgery. AB - Image guidance in pituitary surgery has evolved since diagnostic imaging of the sellar region was first introduced at the turn of the 20th century. These advances have played a key role in the decrease in morbidity and mortality once associated with pituitary surgery. This chapter details the history of sellar imaging as a preoperative diagnostic aide, and then examines the subsequent development of image guidance systems and intraoperative imaging. The utility and limitations of common intraoperative aides including video fluoroscopy, frameless stereotaxy, ultrasound, and magnetic resonance imaging are reviewed. PMID- 16474216 TI - Endoscopic endonasal cavernous sinus surgery, with special reference to pituitary adenomas. AB - Cavernous sinus surgery has always been a surgical challenge because of the high functional importance of this region and the associated high morbidity. The augmented peripheral vision of the endoscope has led to the development of surgical approaches that allow adequate exposure of the cavernous sinus, with a reduction in surgical morbidity. Since 1998, 65 patients with pituitary adenomas and intraoperative evidence of cavernous sinus invasion were treated with a purely endoscopic approach. Follow-up was of at least 6 (mean 51.2) months. There was no perioperative mortality and extremely low morbidity. Radical tumor removal was obtained in 21/35 cases with nonfunctioning adenomas. Hormonal remission was obtained in 13/30 functioning adenomas. One patient with partial hypopituitarism and 1 patient with persistent diabetes insipidus were seen. Three patients with delayed CSF leaks required endoscopic repair. In 1 patient with hemorrhagic infarction in a residual tumor, reintervention with craniotomy was necessary. We advocate the central role of surgery in the treatment of cavernous sinus tumors, since it allows definition of true cavernous sinus involvement, histopathological diagnosis and, when cure is not feasible, tumor volume reduction, which might be an important factor in the response to adjuvant therapy. PMID- 16474217 TI - Diagnosis and management of pediatric sellar lesions. AB - Pituitary region tumors in pediatric patients are largely comprised of craniopharyngiomas and pituitary adenomas, each with their unique considerations. Craniopharyngiomas account for the majority of pediatric sellar masses. Pituitary adenomas are relatively uncommon during childhood, although the incidence increases during adolescence. The diagnosis of sellar lesions involves a multidisciplinary effort, and detailed endocrinologic, ophthalmologic and neurologic testing are critical. The management of pituitary tumors varies depending on the entity. For most tumors, other than prolactinomas, transsphenoidal resection remains the mainstay of treatment. Less invasive modalities such as endoscopic transsphenoidal surgery, and stereotactic radiosurgery have shown promise as primary and adjuvant treatment modalities, respectively. PMID- 16474218 TI - The craniopharyngioma. AB - The craniopharyngioma is one of the most common destructive lesions of the hypothalamus and pituitary gland. It still remains one of the most difficult tumors to treat effectively since complete resection is often impossible and is associated with frequent recurrence. Current therapy is multimodal and focuses on a combination of surgical decompression, medical treatment, as well as stereotactic radiosurgery. This chapter reviews the embryology, neuroanatomy, current treatment strategies, clinical features and the several surgical approaches to its treatment. PMID- 16474219 TI - Rathke's cleft cysts. AB - Rathke's cleft cysts are typically regarded as benign cystic lesions of the sella that may affect the pituitary gland and on occasion the visual apparatus. They are most commonly incidental and rarely of clinical significance. As medical neuroimaging and surgical technologies have rapidly advanced, so too has the discovery, experience, knowledge, and intrigue concerning this relatively rare disease entity. Nevertheless, numerous controversies still exist regarding its natural history, recurrence rate, predictive variables, and optimal surgical management. This chapter aims to review the pathogenesis, symptomatologic manifestations, radiographic, morphologic and histopathological characteristics, treatment strategies and outcomes in the cysts of Rathke's cleft. PMID- 16474220 TI - Treatment of Cushing's disease: a retrospective clinical study of the latest 100 cases. AB - We evaluate the current role of microsurgery for Cushing's disease (CD) and the efficacy of adjuvant treatment modalities. The standard treatment for primary CD remains transsphenoidal surgery followed by adjuvant therapy in cases with persisting hypercortisolism. Moderately severe cases are treated with radiotherapy, while in the very severe adrenalectomy is performed. In our series of primary CD (March 1997 to September 2004, mean observation period 18.8 months) adenomas were confirmed intraoperatively in 84.0% of the cases. Remission was achieved in 75.0% and recurrence was observed in 4.8% of the patients. Complications occurred in 2.0% of the cases and all resolved without resulting in permanent morbidity. In the literature, the rates of intraoperative confirmation of an adenoma vary between 59.1 and 100%, remission rates between 42 and 100%, and recurrence rates between 3.0 and 63.2% depending on the experience of the surgeon and on the definition of remission. These rates have not improved significantly over the years. In experienced hands selective adenomectomy remains the least damaging and most effective treatment modality since it results in rapid clinical improvement if performed successfully. Therefore, it remains the treatment of choice. Patients not cured by surgery alone benefited from a combination of adjuvant treatment tailored to their specific needs using medications, radiation and/or adrenalectomy. In this fashion, we achieved normalization of cortisol levels in 79% and improvement in another 18% of the patients. We expect these rates to increase further once patients treated with radiotherapy begin to experience its full effect within the next few years. PMID- 16474221 TI - Stereotactic radiosurgery for pituitary adenomas: a review of the literature and our experience. AB - Pituitary adenomas are not just one of the most common intracranial tumors but also one of the most difficult to cure. Neurosurgeons have adapted their tools to include precise ionizing radiation in the form of the gamma knife to treat pituitary adenomas. The use of the gamma knife in the management of pituitary adenomas following microsurgery or in selected cases as a primary treatment is safe. The combined application of transsphenoidal surgery and Gamma Knife surgery is beneficial in many difficult cases. However in some patients, optimal control of tumor growth and normalization of hypersecretory states are not achieved. Innovative improvements in operative and radiosurgical techniques are required to avoid pituitary insufficiency and to reduce the number of the cases in which optimal radiosurgery is not feasible because of close tumor proximity to the optic pathways. PMID- 16474222 TI - Neuropathological considerations of pituitary adenomas. AB - Pituitary tumors constitute around 15-20% of intracranial tumors. The understanding of the molecular mechanisms of tumorigenesis and the functional regulation of pituitary adenoma has greatly advanced in the last decade. The current WHO classification scheme of pituitary tumors reflects this progress on tumor biology knowledge, and embraces the most widely utilized diagnostic methods of evaluation of these lesions. The present chapter highlights the different aspects of the tumor diagnosis and reviews the classification of pituitary tumors. PMID- 16474223 TI - Anesthetic and critical care management of patients undergoing pituitary surgery. AB - Patients with tumors of the pituitary gland represent a heterogeneous yet commonly encountered neurosurgical population. Optimal anesthetic care requires an understanding of the complex pathophysiology secondary to each patient's endocrinological disease. Although patients presenting with Cushing's disease and acromegaly have unique manifestations of endocrine dysfunction, all patients with tumors of the pituitary gland require meticulous preoperative evaluation and screening. There are many acceptable strategies for optimal intraoperative anesthetic management; however, the selection of anesthetic agents should be tailored to facilitate surgical exposure, preserve cerebral perfusion and oxygenation, and provide for rapid emergence and neurological assessment. A rapid emergence from anesthesia is particularly important, as an early neurological assessment is necessary to evaluate cranial nerve integrity. In the postoperative period, careful monitoring of fluid balance and serum sodium is essential to the early diagnosis of diabetes insipidus (DI) and the syndrome of inappropriate anti diuretic hormone (SIADH) secretion. DI is most often transient, but can require medical therapy. SIADH has a number of treatment options and decisions should be based upon the patient's status. A thorough understanding of the preoperative assessment, intraoperative management, and potential complications is fundamental to successful perioperative patient care and avoidance of morbidity and mortality. PMID- 16474224 TI - Vascular injury and transsphenoidal surgery. AB - Vascular complications can and do arise from transsphenoidal surgery and, when they occur, they have a high incidence of mortality and serious morbidity. The anatomic substrate for such complications is discussed, along with technical aspects of surgery and other methods for the avoidance of vascular complications. PMID- 16474225 TI - Comparative study of antimicrobial release kinetics from polymethylmethacrylate. AB - Polymethylmethacrylate loaded with antimicrobial agents (most commonly vancomycin and/or aminoglycosides) is used for treatment and prevention of orthopaedic infections. Emergence of organisms resistant to vancomycin or aminoglycosides or both has been reported. Therefore, we studied in vitro release from polymethylmethacrylate beads of antimicrobials with suitable spectra for orthopaedic infections, including cefazolin, ciprofloxacin, gatifloxacin, levofloxacin, linezolid, and rifampin (2.5%, 7.5%, and 15%). Beads were placed in a continuous flow chamber, and antimicrobial concentrations in chamber outflow were determined by bioassay at timed intervals thereafter. Release profiles were bimodal with initial rapid release of high concentrations followed by sustained, slow release. Antimicrobial agents studied showed varied release profiles, indicating that elution from polymethylmethacrylate is unique to individual antimicrobial agents. Increasing antimicrobial concentration in polymethylmethacrylate increased peak concentrations and area under the curve. Cefazolin, ciprofloxacin, gatifloxacin, levofloxacin, linezolid, and rifampin may be suitable for incorporation into polymethylmethacrylate for management of orthopaedic infections. PMID- 16474227 TI - Clinical experience and progress of computed tomographic coronary angiography. AB - Multidetector computed tomography is rapidly emerging as a noninvasive method to evaluate the coronary anatomy. Technical advances available in modern scanners now lead to adequate image quality in most patients. Image acquisition and interpretation can be performed very rapidly, making this technology suitable for the evaluation of ambulatory patients. This review summarizes the capabilities and limitations of multidetector computed tomography coronary angiography performed with current generation scanners. PMID- 16474228 TI - Advantages of multidetector computed tomography angiography in the evaluation of patients with chest pain. AB - Noninvasive imaging of the coronary arteries using multidetector computed tomography is gaining recognition among clinicians. The rapid evolution of this technology is opening new frontiers in the diagnosis of coronary artery disease. This article reviews the current status and future potential for the application of noninvasive coronary multidetector computed tomography angiography in the assessment of coronary artery stenosis, coronary artery anomalies, mural plaque and coronary stent and bypass graft patency. PMID- 16474229 TI - Cardiac imaging using multislice computed tomography scanners: technical considerations. AB - Conventional coronary angiography is currently the gold standard in the detection and diagnosis of coronary artery disease. This modality, however, is invasive in nature. Hence, there is a need for noninvasive imaging techniques to provide comprehensive assessment of coronary artery disease, especially in stable patients at low to moderate risk of disease. In recent years, a number of noninvasive modalities have found wide applications in cardiac imaging. Most recent investigations have used magnetic resonance imaging, multislice computed tomography and electron-beam computed tomography scanners. This review discusses the clinical challenges existing in the field of cardiac imaging and focuses on the technical advancements of multislice computed tomography scanners that have made them a very attractive noninvasive option for the detection and diagnosis of coronary artery disease. PMID- 16474230 TI - Multidetector computed tomography guidance in complex cardiac ablations. AB - Radiofrequency ablation of complex cardiac arrhythmias has undergone significant evolution in the past decade, with the development of technology enabling better anatomic and electrophysiologic mapping of abnormal cardiac tissue. In this paper, we will discuss the role of pre-procedural and post-procedural multidetector computed tomography, with specific focus on the anatomic assessment of pulmonary vein and left atrial anatomy in the ablation of atrial fibrillation. We will also consider how the integration of both multidetector computed tomography and electroanatomic computer-based imaging may contribute more broadly to the management of a variety of complex ablation procedures. PMID- 16474231 TI - Predictors of long-term outcome following percutaneous coronary intervention in patients with type 2 diabetes mellitus. AB - OBJECTIVES: Diabetes has a major impact on the long-term outcome following percutaneous coronary intervention, being a major risk factor for restenosis. We aimed at assessing the clinical, hemodynamic and angiographic factors that predict outcome in this high-risk group. METHODS: Diabetic patients who underwent percutaneous coronary intervention during the period 1996-2000 were identified retrospectively, and their medical records, hemodynamic data and angiograms studied at the time of their procedure. Angiographic data were analyzed to determine procedural factors, quantitative angiographic results of their percutaneous coronary intervention and severity and extent scores of coronary artery disease. Clinical follow-up was obtained in 99.7% to a mean time of 782+/ 153 days after percutaneous coronary intervention. RESULTS: A total of 327 consecutive diabetic patients underwent percutaneous coronary intervention on 389 lesions. The overall procedural success rate was 96.2%; however, during follow up, 40.2% of patients had an adverse cardiac event. The strongest independent predictors of adverse outcome were presence of renal disease, need for medical diabetic treatment, and the extent of coronary artery disease as assessed by the extent score, but not the duration of diabetes or glycemic control. Other factors, which are often predictive of short-term outcome in a general population, such as vessel diameter, lesion length, and severity of coronary artery disease, were not predictive of outcome. CONCLUSIONS: In patients with type 2 diabetes mellitus, the extent of the coronary arteries involved with atheroma as well as systemic factors such as renal involvement are the best predictors of long-term outcome following percutaneous coronary intervention. PMID- 16474232 TI - Pressure-derived collateral flow index: a strong predictor of late left ventricular remodeling after thrombolysis for acute myocardial infarction. AB - OBJECTIVE: Despite proved efficacy of pressure-derived collateral flow index in determining microvascular dysfunction in patients with acute myocardial infarction, its role in prediction of left ventricular remodeling at long term has yet to be demonstrated. In this study, we investigated the relationship between quantitatively assessed microvascular dysfunction by using intracoronary pressure wire and late left ventricular remodeling. PATIENTS AND METHODS: The study population consisted of 28 patients with first acute myocardial infarction. They were treated with fibrinolytic therapy. The inclusion criteria were thrombolysis in myocardial infarction grade II-III flow in infarct-related artery and all destined for stent implantation. Cardiac catheterization and stent implantation were performed in mean of 3.3 days after acute myocardial infarction. During the stent implantation procedure, the pressure-derived collateral flow index was measured by using intracoronary pressure wire. Control angiograms were performed at 6+/-2 months. Echocardiographic left ventricular volume indexes were measured at discharge, at 6 months and at 1 year. Changes in left ventricular volumes from baseline to 1 year were followed. RESULTS: Left ventricular end-diastolic volume index at 1 year correlated significantly with the pressure-derived collateral flow index (r=0.69, P<0.01). A significant correlation was also observed between the change in left ventricular end diastolic volume index from baseline to 1 year and the pressure-derived collateral flow index (r=0.65, P<0.01). The most important predictor of 1-year left ventricular remodeling was the pressure-derived collateral flow index (P<0.0001), and collateral circulation (P=0.03). CONCLUSION: The pressure-derived collateral flow index is a powerful independent predictor of 1-year left ventricular dilatation. Given its simplicity of measurement, and correlation with microvascular obstruction and left ventricular outcome at long term, the pressure derived collateral flow index may provide useful and valuable estimates of clinical outcomes after acute myocardial infarction. PMID- 16474233 TI - Cardiovascular calcifications after radiation therapy for Hodgkin lymphoma: computed tomography detection and clinical correlation. AB - OBJECTIVE: To study cardiovascular calcifications, detected by computed tomography, in patients following mediastinal radiation for Hodgkin lymphoma, and correlate them with clinical findings. MATERIALS AND METHODS: Fifteen patients, or=30 mg/dl than in patients with levels <30 mg/dl (72 vs. 37%, P=0.008). A strong negative correlation was observed between lipoprotein (a) and vascular endothelial cell growth, factor (r= 0.708, P<0.0001). Multivariate analysis revealed that a high level of lipoprotein (a) negatively affected the development of collaterals, whereas the duration of angina had a positive effect. CONCLUSION: This study demonstrated for the first time that the high level of lipoprotein (a) negatively affects the formation of coronary collateral vessels in human beings. Reduced production or bioactivity of vascular endothelial cell growth factor caused by high levels of lipoprotein (a) may be the possible responsible mechanisms of hyperlipoprotein (a)-related poor collateral formation. PMID- 16474236 TI - Role of right-sided chest leads in the detection of multivessel coronary artery disease in patients with extended Q-wave anterior myocardial infarction. AB - OBJECTIVE: To evaluate the improvement of diagnostic ability of exercise testing to detect multivessel coronary artery disease in patients with extended Q-wave anterior myocardial infarction, using additional right-sided chest leads. METHODS: Fifty-two consecutive patients with Q-wave anterior myocardial infarction underwent exercise testing, using the standard 12 and the additional right-sided (V3R, V4R, V5R) chest leads, thallium-201 scintigraphy and coronary arteriography. RESULTS: Twenty-one (40%) patients had one-vessel disease, 18 (35%) had two-vessel disease and 13 (25%) had three-vessel disease. The sensitivities of the standard 12-lead exercise testing and its combination with the additional right-sided chest leads were 24% (5/21) versus 28% (6/21) for the detection of one-vessel disease (P: NS), 33% (6/18) versus 83% (15/18) for the detection of two-vessel disease (P<0.05) and 38% (5/13) versus 92% (12/13) for the detection of three-vessel disease (P<0.05), respectively. In thallium-201 scintigraphy, 29 of the 31 (94%) patients with multivessel coronary artery disease demonstrated reversible ischemia. The usual 12-lead exercise testing could detect ischemia in 11 (35%) of these 31 patients, while the addition of the right-sided chest leads could detect ischemia in 27 (87%) of them (P<0.05). CONCLUSIONS: The additional right-sided chest leads significantly improve the low sensitivity of the usual exercise testing to detect multivessel coronary artery disease in patients with previous extended Q-wave anterior myocardial infarction. PMID- 16474237 TI - A meta-analysis of safety and effectiveness of perioperative beta-blocker use for the prevention of cardiac events in different types of noncardiac surgery. AB - OBJECTIVE: Perioperative beta-blocker therapy has been proposed to improve outcome. Most of the trials conducted, however, lacked statistical power to evaluate the incidence of hard cardiac events and the relationship to the type of surgery. Therefore, we conducted a meta-analysis of all randomized controlled trials in which beta-blocker therapy was evaluated. METHODS: An electronic search of published reports on Medline was undertaken to identify studies published between January 1980 and November 2004 in English language journals. All studies reported on at least one of three endpoints: perioperative myocardial ischemia, perioperative nonfatal myocardial infarction, and cardiac mortality. Type of surgery, defined as low, intermediate, and high risk according to the American College of Cardiology/American Heart Association guidelines, was noted. RESULTS: In total, 15 studies were identified, which enrolled 1,077 patient. No significant differences were observed in baseline clinical characteristics between patients randomized to beta-blocker therapy and control/placebo. Beta blocker therapy was associated with a 65% reduction in perioperative myocardial ischemia (11.0% vs. 25.6%; odds ratio 0.35, 95% confidence interval 0.23-0.54; P<0.001). Furthermore, a 56% reduction in myocardial infarction (0.5% vs. 3.9%, odds ratio 0.44, 95% confidence interval 0.20-0.97; P=0.04) and a 67% reduction (1.1% vs. 6.1%, odds ratio 0.33, 95% confidence interval 0.17-0.67; P=0.002) in the composite endpoint of cardiac death and nonfatal myocardial infarction were observed. No statistical evidence was observed for heterogeneity in the treatment effect in subgroups according to type of surgery (P for heterogeneity 0.2). CONCLUSION: This meta-analysis shows that beta-blocker use in noncardiac surgical procedures is associated with a significant reduction of perioperative cardiac adverse events. PMID- 16474238 TI - Coronary pressure measurement to identify the lesion requiring percutaneous coronary intervention in equivocal tandem lesions. AB - OBJECTIVES: No reliable methods are available for determining application of percutaneous coronary intervention for treatment of equivocal tandem lesions. We investigated whether coronary pressure measurement is useful for determining the lesion that requires percutaneous coronary intervention in tandem lesions. METHODS: We measured coronary pressure in 72 consecutive patients with tandem lesions. Myocardial fractional flow reserve (FFRmyo) was obtained as the ratio of coronary pressure distal to the lesion/aortic pressure under maximal hyperemia. If the FFRmyo across the tandem lesions was >or=0.75, we deferred percutaneous coronary intervention for the lesion. When the tandem lesions showed FFRmyo<0.75, percutaneous coronary intervention was performed on the lesion that showed angiographically higher stenosis. When FFRmyo was <0.75 after one-lesion percutaneous coronary intervention, this intervention was carried out on the remaining lesion. RESULTS: We deferred percutaneous coronary intervention for 26 patients (36.1%), and performed percutaneous coronary intervention in 46 patients (63.8%). We performed percutaneous coronary intervention for one lesion in 19 patients (26.4%) and for both lesions in 27 patients (37.5%). Among patients in whom percutaneous coronary intervention was deferred, only two patients (7.7%) required target lesion revascularization during the follow-up period. This rate was not higher than that in the 46 patients who underwent percutaneous coronary intervention for one or two lesions (six patients, 13.0%). Similarly, the target lesion revascularization in lesions with initially deferred percutaneous coronary intervention (5.6%, 4/71 lesions) was not higher than that in lesions with percutaneous coronary intervention (15.1%, 11/73 lesions). Major cardiac events, cardiac death and acute myocardial infarction, did not occur in patients with deferred percutaneous coronary intervention and in those with percutaneous coronary intervention during the follow-up period. CONCLUSION: Our results clearly showed that coronary pressure measurement was clinically useful for identifying equivocal tandem lesions requiring percutaneous coronary intervention. PMID- 16474239 TI - Bibliography. Current world literature. PMID- 16474240 TI - Change is hard--how many times have we heard that saying? PMID- 16474242 TI - Certification of the holistic nurse. PMID- 16474243 TI - Medication management educational tools for improving target outcomes: any emergent care. AB - United Home Health Services, Inc. selected the utilization outcome referred to as "any emergent care" provided as our target focus for 2004/2005. After the process of care investigation and development of important care behaviors, two medication management educational tools were generated. Educational in-services were provided to clinical staff. PMID- 16474244 TI - Correlation among client satisfaction, nursing perception of outcomes, and organizational variables. AB - Client satisfaction as an outcome measure provides useful information to home healthcare managers. After a review of client satisfaction instruments in home healthcare, a study of 38 home health sites indicated that clients had a high level of satisfaction with nursing care. A correlational descriptive study compared the client satisfaction to the Community Health Nursing Outcomes Inventory. Findings suggest that client satisfaction was higher when nurses perceive meeting clients' psychosocial needs, when nurses and supervisors maintain a good working relationship, and when nurses use guidelines to direct nursing practice. PMID- 16474246 TI - Teamwork keeps the pressure off: the role of the occupational therapist in the prevention of pressure ulcers. PMID- 16474247 TI - Falls prevention, or "I think I can, I think I can": an ensemble approach to falls management. AB - Home care clinicians face a daunting challenge in attempting to prevent falls among the frail, elderly population. This article outlines how one agency used evidence-based strategies to develop a program that identified patients at risk for falls and intervened with the goal of decreasing injury from falls by using a multidisciplinary approach. PMID- 16474248 TI - Caring for wounds: more than just cuts and scrapes. PMID- 16474249 TI - Wound care in the face of life-limiting illness. PMID- 16474251 TI - Do you know where your critical lab values are?: communication of critical lab results. PMID- 16474252 TI - "Ask Home Healthcare Nurse" (August 2005). PMID- 16474253 TI - Mesenteric blood flow velocities in the newborn with single-ventricle physiology: modified Blalock-Taussig shunt versus right ventricle-pulmonary artery conduit. AB - BACKGROUND: Neonates with ductal-dependent single-ventricle congenital heart disease palliated with a modified Blalock-Taussig shunt (mBTS) commonly have retrograde diastolic flow in the aorta, which may place them at increased risk of mesenteric ischemia. Recently, palliation with a right ventricle-to-pulmonary artery conduit, known as the Sano procedure, has been shown to eliminate retrograde diastolic flow, theoretically leading to better systemic perfusion. OBJECTIVE: To compare the changes in superior mesenteric artery (SMA) and celiac artery velocities and flow after a bolus enteral feed in patients with single ventricle congenital heart disease palliated with an mBTS vs. those palliated with the right ventricle-to-pulmonary artery conduit. DESIGN: Prospective clinical study. SETTING: Cardiothoracic intensive care unit and pediatric ward of a tertiary care children's hospital. PATIENTS: A total of 27 patients with single ventricle congenital heart disease (15 with mBTS, 12 with Sano) after stage-1 palliation. INTERVENTION: Doppler ultrasound of the SMA and celiac artery was performed 30 mins before and after a bolus enteral feed. MEASUREMENTS AND MAIN RESULTS: SMA and celiac artery peak systolic flow velocity, mean flow velocity, and time-velocity integral were measured. After a bolus enteral feed, 8 of 15 infants palliated with an mBTS had retrograde diastolic flow through the SMA yet demonstrated significant increases in all variables of both the SMA and celiac artery flow velocities (SMA peak systolic flow velocity: 0.96 +/- 0.33 vs. 1.2 +/ 0.4 m/sec, p = .01). Those palliated with the Sano procedure did not demonstrate SMA retrograde diastolic flow but also did not have any significant changes in their mesenteric flow variables (SMA peak systolic flow velocity: 0.79 +/- 0.16 vs. 0.89 +/- 0.26 m/sec, p = .2). CONCLUSION: Postprandial retrograde diastolic flow was observed in the majority of patients palliated with an mBTS vs. none of the patients in the Sano group. However, contrary to expectations, postprandial mesenteric blood flow velocities in those palliated with an mBTS are significantly higher than in Sano patients, although the increase is not as high as that historically seen in normal neonates. This may place this population at risk for mesenteric ischemia and feeding intolerance in the postoperative period, and the risk may be even greater for those neonates palliated with a right ventricle-to-pulmonary artery conduit. PMID- 16474254 TI - Heparin-induced thrombocytopenia in a pediatric patient receiving extracorporeal support and treated with argatroban. AB - INTRODUCTION: Heparin-induced thrombocytopenia (HIT) is rare in the pediatric population, with a majority occurring in the pediatric intensive care unit setting. All reported cases have been associated with the use of unfractionated heparin. Because unfractionated heparin is the anticoagulant of choice for extracorporeal life support, the development of HIT in these patients can be devastating. We report a case of HIT with evidence of small-vessel arterial thromboembolism in a 17-month-old child receiving extracorporeal membrane oxygenation and continuous renal replacement therapy successfully treated with argatroban. CASE: The patient was a 17-month-old boy with severe respiratory failure secondary to asthma and mucus plugging that failed conventional and unconventional ventilation. Venovenous extracorporeal membrane oxygenation was initiated, and within 24 hrs, there was a precipitous decrease in the platelet count, with the development of cutaneous ischemia involving his lower limbs. Heparin-associated antibodies were positive. Argatroban was started, and the child maintained on extracorporeal membrane oxygenation and continuous renal replacement therapy, with resolution of the cutaneous ischemia and rebound of the thrombocytopenia. DISCUSSION: HIT is rare in the pediatric population. Recognition of HIT is vital because withdrawal of heparin is the first and most important therapy. For patients on extracorporeal membrane oxygenation or continuous renal replacement therapy who develop HIT, synthetic thrombin inhibitors (hirulogs) have been reported as an alternative. However, little information on their use in extracorporeal life support has been published, particularly in the pediatric population. CONCLUSION: This report documents a pediatric case of HIT successfully treated with argatroban, allowing continuation of the venovenous extracorporeal membrane oxygenation and continuous renal replacement therapy, with resolution of the thromboembolic ischemia and thrombocytopenia. PMID- 16474255 TI - Early dynamic changes in pulse oximetry signals in preterm newborns with histologic chorioamnionitis. AB - OBJECTIVE: No reliable clinical markers of histologic chorioamnionitis (HCA), a major and often subclinical cause of prematurity leading to high neonatal morbidity and mortality, are available to date. Increasing evidence indicates myocardial dysfunctions in affected fetuses and newborns. We sought to assess the value of nonlinear dynamics from pulse oximetry signals in identifying affected newborns. DESIGN: Prospective case-control study. SETTING: Tertiary level neonatal intensive care unit, Brindisi Hospital. PATIENTS AND INTERVENTION: Pulse oximetry-derived signals (pulse rate, oxygen saturation, and perfusion index), recorded within the first 1.5 hrs of life, were analyzed for 110 very low-birth weight infants, of whom 54 had histopathological evidence of HCA. MEASUREMENTS AND MAIN RESULTS: Four different time series parameters were determined for nonlinear dynamical (NLD) analysis. Significantly decreased Lempel-Ziv, Lyapunov largest exponent, and correlation dimension, with significantly increased Hurst values for heart rate and perfusion index (p < .00001), were observed in newborns with HCA. Heart rate Lempel-Ziv 7 days. The control group included all patients admitted for >7 days who needed mechanical ventilation from January 1, 1994, to December 31, 1997. INTERVENTIONS: Oral amphotericin B suspension, 50 mg every 8 hrs, administered to all study group patients soon after initiation of mechanical ventilation and terminating after weaning. MEASUREMENTS: The rates of Candida bloodstream infection were compared between the study and control groups. MAIN RESULTS: Candida species were isolated from blood cultures in 5 of 185 (2.1%) and 21 of 196 (10.7%) patients in the study and control groups, respectively (p= .0038). There was also a statistically significant (p= .017) decrease in Candida bloodstream infection rate in all patients admitted to the pediatric intensive care unit for >7 days during the study period compared with the Candida bloodstream infection rate during the control period. CONCLUSION: Prophylactic administration of oral amphotericin B may lead to a significant decrease in the rate of Candida bloodstream infection in ventilated pediatric intensive care unit patients. PMID- 16474260 TI - The beryllium occupational exposure limit: historical origin and current inadequacy. PMID- 16474261 TI - Morbidity ranking of U.S. workers employed in 206 occupations: the National Health Interview Survey (NHIS) 1986-1994. AB - OBJECTIVE: The objective of this study was to rank U.S. occupations by worker morbidity. METHODS: From 1986 through 1994, morbidity information was collected on over 410,000 U.S. workers who participated in the National Health Interview Survey, an annual household survey representative of the U.S. civilian noninstitutionalized population. A multivariate adjusted logistic regression morbidity summary score was created for each worker group based on seven indicators: days of restricted activity, bedrest, and missed work in the previous 2 weeks; doctor visits and hospitalizations in the previous 12 months; reported health conditions; and health status. RESULTS: Worker groups reporting the greatest morbidity included social workers, inspectors, postal clerks, psychologists, and grinding machine operators; worker groups reporting the least morbidity included dentists, pilots, physicians, pharmacists, and dietitians. CONCLUSIONS: These findings aid in the identification of worker groups that require increased attention for morbidity research and prevention. PMID- 16474262 TI - The impact of rheumatoid arthritis on medical expenditures, absenteeism, and short-term disability benefits. AB - OBJECTIVES: The objectives of this study were to estimate medical expenditures, absenteeism, and short-term disability costs for workers with rheumatoid arthritis (RA) and to estimate the relative costs of RA over a 12-month period. METHODS: Using data from nine U.S. employers, direct and indirect costs for 8502 workers with RA were compared with costs for a matched group without RA. Regression analyses controlled for factors that were different even after propensity score matching. RESULTS: Average total costs for workers with RA were $4244 (2003 dollars) greater than for workers without RA. RA was the fourth most costly chronic condition per employee compared with cancers, asthma, bipolar disorder, chronic obstructive pulmonary disease, depression, diabetes, heart disease, hypertension, low back disorders, and renal failure. CONCLUSIONS: RA is a costly disorder and merits consideration as interventions are considered to improve workers' health and productivity. PMID- 16474263 TI - Evaluation of flavorings-related lung disease risk at six microwave popcorn plants. AB - OBJECTIVE: After investigating fixed airways obstruction in butter flavoring exposed workers at a microwave popcorn plant, we sought to further characterize lung disease risk from airborne butter-flavoring chemicals. METHODS: We analyzed data from medical and environmental surveys at six microwave popcorn plants (including the index plant). RESULTS: Respiratory symptom and airways obstruction prevalences were higher in oil and flavorings mixers with longer work histories and in packaging-area workers near nonisolated tanks of oil and flavorings. Workers were affected at five plants, one with mixing-area exposure to diacetyl (a butter-flavoring chemical with known respiratory toxicity potential) as low as 0.02 ppm. CONCLUSIONS: Microwave popcorn workers at many plants are at risk for flavoring-related lung disease. Peak exposures may be hazardous even when ventilation maintains low average exposures. Respiratory protection and engineering controls are necessary to protect workers. PMID- 16474264 TI - Plasma mutant-p53 protein and anti-p53 antibody as a marker: an experience in vinyl chloride workers in Italy. AB - OBJECTIVE: The objective of this study was to examine the presence of mutant-p53 protein (p53Ag) and antibodies to p53 protein (p53Ab) in a population of workers exposed to vinyl chloride (VC). METHOD: We have investigated the presence of two cancer markers in the plasma of 151 subjects exposed to varying concentrations of VC (4-2823 ppm). The investigation took place in two sessions: in 1999, the analysis was limited to p53Ab, and in 2000, the analysis was repeated and extended to include also the mutant-p53Ag. The available information on the subjects in this study includes age, total years of employment in the VC polymerization industry, exposure concentration, results of abdominal ultrasonogram, hepatitis status, smoking and alcohol drinking status, and clinical records. Logistic regression analyses were performed to calculate the association between prevalence of positivity for p53Ab or mutant-p53Ag and cumulative VC exposure concentration after adjustment for confounding factors. t test and chi analyses were performed to test significant differences among groups. RESULTS: Three (1.9%) of the 151 workers exposed to VC resulted seropositive for the mutant-p53Ag and five (3.3%) for the p53Ab. All seropositive subjects are distributed in the highest exposure classes (>1000 ppm). No seropositivity was found among controls. The stratified relationship between seropositivity and exposure appeared statistically significant (chi = 23.65 for mutant-p53Ag and 30.35 for p53Ab). CONCLUSIONS: revealing its presence in subjects having a history of VC exposure greater than 1000 ppm. On the basis of this study, and the clinical experience of the authors, the presence of a minimum threshold for the carcinogenic effects of VC is hypothesized. PMID- 16474265 TI - Parental occupational exposure and the risk of acute lymphoblastic leukemia in offspring in Israel. AB - OBJECTIVE: Parental employment in occupations that have potential exposures to organic solvents or pesticides could be associated with the risk of childhood acute lymphoblastic leukemia (ALL) in their offspring. METHODS: We explored this hypothesis by studying the association with respect to exposure time windows. Our case-control study included 224 children, 112 diagnosed with ALL and 112 matched controls. RESULTS: A significantly higher odds ratio (OR) was found between childhood ALL and reported parental occupational exposures. Analysis of exposures of both parents by exposure time windows revealed significant OR during the preconception and postnatal periods separately. CONCLUSIONS: The results provide support to the association between parental occupational exposures and ALL in their children. These results should be interpreted cautiously because of the small numbers, biases characterizing case-control studies, and the use of hospital-based controls. PMID- 16474266 TI - Effect of styrene on postural stability among reinforced plastic boat plant workers in Finland. AB - OBJECTIVE: Many drugs are vestibulotoxic and may impair balance. Styrene is both ototoxic and neurotoxic, but its effect on balance has been little studied. We evaluate the effect of low concentrations of styrene on balance among Finnish fiberglass-reinforced plastic boat manufacturers. METHODS: From 252 male employees, we measured postural stability and urine mandelic and phenylglycolic acid concentrations. Individual exposure to styrene in breathing zone was measured for 148 workers. For analysis, 88 matched pairs were selected. RESULTS: The postural stability of laminators was aggravated compared with nonlaminators. This impairment was observed in young workers, and it worsened with age. CONCLUSIONS: This study documents styrene as an additional risk factor in impaired postural stability, with the negative effects of the styrene exposure starting already in younger workers. PMID- 16474268 TI - Evaluation of occupational exposure to N-nitrosamines in a rubber-manufacturing industry. AB - OBJECTIVE: We sought to determine volatile N-nitrosamines levels in the air of a rubber-manufacturing industry and to measure urinary N-nitrosamines concentrations in exposed workers. METHODS: Personal monitoring of 34 workers was performed by sampling nine airborne N-nitrosamines in four factories that manufactured rubber drive belts for automotive engines. Urinary N-nitrosamine levels were determined in all workers and in a control group of 26 subjects. Analyses were conducted by capillary gas chromatography-thermal energy analyzer. RESULTS: Airborne and urinary N-nitrosamines levels were very low and, in most cases, below the limit of detection (0.06 microg/m and 0.1 microg/L, respectively). CONCLUSIONS: Although airborne and urinary N-nitrosamines levels were found to be very low, exposed workers should still be monitored constantly because some of these substances are known to be genotoxic and carcinogenic. PMID- 16474267 TI - Correlating atmospheric and biological markers in studies of secondhand tobacco smoke exposure and dose in children and adults. AB - OBJECTIVE: We sought to directly compare secondhand smoke (SHS) atmospheric markers to each other and to SHS dosimetric biomarkers, permitting intercomparison of clinical and atmospheric studies. METHODS: We used atmospheric and pharmacokinetic (PK) models for the quantitative estimation of SHS exposure and dose for infants, children, and adults, based on building smoker density and air exchange rate, and from exposure duration, default PK parameters, and respiration rates. RESULTS: We estimate the SHS serum cotinine doses for the typical and most-exposed individuals in the U.S. population; predictions compare well to measurements on a national probability sample. Using default respiration rates, we estimate serum cotinine dose from SHS nicotine exposure for 40 adults exposed to SHS in an environmental chamber; predictions agreed with observations. We correlate urine cotinine and hair nicotine levels for 127 infants exposed to parental smoking, and estimate corresponding atmospheric nicotine exposure via PK modeling. CONCLUSIONS: Our "Rosetta Stone" Equations allow the SHS atmospheric markers, respirable particles, nicotine, and carbon monoxide, to be related to the SHS biomarkers, cotinine in blood, urine, and saliva and nicotine in hair, permitting intercomparison of clinical and atmospheric studies of SHS for the first time. PMID- 16474269 TI - Respiratory cancer risks among workers with glutaraldehyde exposure. AB - OBJECTIVE: Glutaraldehyde is a substance that represents a substantial portion of the human exposure to aldehydes in medicine and industry. Other aldehydes such as formaldehyde have been associated with increased cancer rates of the upper respiratory tract and leukemia. Our study is the only one, to our knowledge, to examine cancer rates among exposed glutaraldehyde workers. METHODS: In an extended follow up using death certificates, we calculated standardized mortality ratios (SMRs) and 95% confidence intervals (CIs) for three cumulative exposure categories of glutaraldehyde. There were 99,730 person-years of observation among unexposed workers, 2934 person-years in the lower exposure category, < 0-100.0 parts per billion (ppb)-years, and 2805 person-years in the higher exposure category of 100.0+ ppb-years. RESULTS: For all respiratory cancers for these exposure categories, the SMRs were 0.9 (95% CI = 0.7-1.1), 1.0 (95% CI = 0.2 3.0), and 0.3 (95% CI = 0.0-1.5). No increasing trend of SMR with increasing exposure is observed for any cause of death examined. We observed no cancers of the nasal cavity and sinus (0.03 expected), nasopharynx (0.02 expected), or leukemia (0.6 expected) among all glutaraldehyde-exposed workers. CONCLUSIONS: Although our study findings should be tempered by the small size and the potentially low prevalence of smoking among glutaraldehyde workers, we found no increased rates of respiratory tract cancer or leukemia related to glutaraldehyde exposure. PMID- 16474270 TI - Sensitization and chronic beryllium disease among workers in copper-beryllium distribution centers. AB - OBJECTIVE: Little is known about the risk of sensitization and chronic beryllium disease (CBD) among workers performing limited processing of copper-beryllium alloys downstream of the primary beryllium industry. In this study, we performed a cross-sectional survey of employees at three copper-beryllium alloy distribution centers. METHODS: One hundred workers were invited to be tested for beryllium sensitization using the beryllium blood lymphocyte proliferation test (BeLPT); a sensitized worker was further evaluated for CBD. Available beryllium mass concentration air sampling data were obtained for characterization of airborne exposure. RESULTS: One participant, who had exposure to other forms of beryllium, was found to be sensitized and to have CBD, resulting in a prevalence of sensitization/CBD of 1% for all tested. CONCLUSIONS: The overall prevalence of beryllium sensitization and CBD for workers in these three copper-beryllium alloy distribution centers is lower than for workers in primary beryllium production facilities. PMID- 16474274 TI - [Pathogenesis and virulence determinants of the family Legionellaceae]. AB - Legionellae are Gram-negative obligate intracellular parasites of unicellular animal organisms, some of which are sometimes able to cause an acute and sever pneumonia in humans. Legionellae, are capable of surviving for long periods in water when between hosts. Their adaptation to these quite diverse environments seems to be accomplished by morphological and biochemical pathway changes. It has been well established that humans coexisted with these bacteria for a long period of time, and it is only recently that our industrial technology provided these organisms with the means of causing infection. This review describes the invasion process and the life cycle of Legionellae in both protozoan and mammalian cells. Our understanding of the infection cycle of Legionellae is primarily based on observations by transmission and scanning EM and by contrast phase and fluorescence microscopy. The identification of virulence determinants by molecular techniques is also discussed. The bacteria enter the host cell mainly by coiling phagocytosis and reside within unique phagosomes which, during the first hours of infection, are isolated from the endosomal pathway. Within the protected vacuole the mature infectious forms of Legionellae convert to replicative forms that no longer express virulence traits. Evidence is provided on how the biogenesis of the replication niche is determined. The virulence factors that arrest phagosome maturation during intracellular replication are also described. The status of our current knowledge on the means by which Legionellae successfully infect the host cells to cause disease is discussed. PMID- 16474275 TI - Clusterin: the missing link in the calcium-dependent resistance of cancer cells to apoptogenic stimuli. AB - The resistance of cancer cells to artificially induced apoptosis comprises a major pitfall in contemporary chemotherapy. In recent years, a wide range of molecular mechanisms was revealed that allow cancer cells to overcome apoptosis. In prostate, breast, and colorectal cancers, a protein named clusterin was identified with anti- or proapoptotic activity regulated by calcium homeostasis. Reports so far suggest "two faces" of clusterin activity: the calcium-dependent cellular retention of clusterin positively correlates with cell survival, whereas nuclear translocation of this protein promotes cell death in calcium-deprived cells. Better understanding of the proprieties of clusterin isoforms and the molecular mechanisms which regulate its activity provide the prospect of breaking down cancer cells' resistance to apoptosis. Thus one might expect such dual benefits as overcoming the "immune escape" of neoplastic cells and reducing the doses of cytostatic drugs, with a concomitant reduction in the side effects of chemotherapy. PMID- 16474276 TI - [The role of Toll-like receptors (TLR) in innate and adaptive immune responses and their function in immune response regulation]. AB - The innate immune response is a universal mechanism of host defense against infection. It functions on the basis of special receptors called PRRs (pattern recognition receptors) which recognize conserved microbial structures called PAMPs (pathogen-associated molecular patterns). Due to PRRs, the human organism is able to discriminate between self and non-self antigens. Toll-like receptors (TLRs) are a group of PRRs that play a crucial role in "danger" recognition and the induction of immune response. Cells of the immune system (macrophages, dendritic cells, mast cells, eosinophils, neutrophils, B lymphocytes), epithelial cells, endothelium, cardio-myocytes and adipocytes all recognize pathogens via TLRs. TLR stimulation via microbial products activates the innate immune response. This results in an upregulated synthesis of anti-bacterial substances and pro-inflammatory cytokines as well as the activation of dendritic cell maturation (increased expression of co-stimulatory molecules and MHC antigens), thereby becoming more effective in antigen presentation. In some cases, the innate immune response is not able to eliminate infection and requires the induction of the adaptive immune response. When activated via TLRs, antigen presenting cells (APCs) release elevated levels of pro-inflammatory cytokines (TNF-alpha, IL-1, IL-6, IL-8, and IL-12), chemokines, and nitric oxide (NO) and show increased expression of co-stimulatory molecules (CD40, CD80, CD86). All these changes in APC function allow the induction of the adaptive immune response, where both T and B lymphocytes play a crucial role. TLRs also play a role in the regulation of immune response via direct or indirect influence on the function of CD4+ CD25+ T regulatory cells (Tregs), which results in their induction and subsequent suppression of the immune response or a reversal of suppression (contrasuppression). PMID- 16474277 TI - [The cytoskeleton reorganization and differentiation of HL-60 and K-562 human leukemia cell lines]. AB - Microfilaments, microtubules, and intermediate filaments form the cytoskeleton. These substructures play a significant role in cell motility, transport, divisions, differentiation, tumor transformation, and apoptosis. These processes are related with changes in cell shape, in which cytoskeletal proteins take an active part. In non-muscle cells, actin is an essential constituent of microfilaments, tubulin forms microtubules, and vimentin is one of the characteristic proteins of intermediate filaments. The differentiation of cells is associated inseparably with tissue and organ formation, and the induction of malignant cell differentiation can be a method of treatment, especially in hematopoietic steam cell disease therapy. In studies on tumor cell differentiation, agents such as cytokines, retinoids, forbol esters, and vitamin D3 are the most commonly used, and results show these substances may participate in different pathways of signal transduction. Retinoids and vitamin D3 mostly affect gene transcription via nuclear receptors, whereas cytokines act through membrane receptors. The results of studies show actin, tubulin, and vimentin reorganization during the differentiation of leukemia cells, but it remains unknown whether the observed changes are the cause or the result of the differentiation process. PMID- 16474278 TI - [Implantation and antigenicity of human endometrium]. AB - It is essential for an embryo's further development that it generate a connection with the mother. The first stage of this process is implantation, a highly coordinated event that involves both embryonic and endometrial participation. A blastocyst may attach to the endometrium only during the "implantation window", when the uterus is receptive. A description of the molecular features of a receptive endometrium is the aim of this review. During the secretory phase, the endometrium synthesizes such cytokines as LIF (leukemia inhibitory factor), HB EGF (heparin-binding epithelial growth factor), and TGF-alpha (transforming growth factor alpha) as well as special surface structures, such as integrins. There are two types of integrins: cycle dependent and constitutionally expressed. The coexpression of cycle-dependent integrins may mark the "implantation window". The endometrium of fertile and infertile women may vary in expression of the integrins. Insufficient synthesis of integrins may be caused by a luteal phase deficiency. E-cadherin and alpha-crystalin B play an important role during implantation. The expression of pinopodes on endometrial cells during the "implantation window" marks optimal uterine receptivity. The composition of all the factors mentioned above define the unique properties of the endometrium during the "implantation window". PMID- 16474279 TI - International exchange of residents in physical and rehabilitation medicine: challenges and solutions. PMID- 16474280 TI - The effects of traumatic brain injury on patients and their families. A follow-up study. AB - AIM: With this study we wanted to verify whether the same uneasiness elements as those described by numerous previous studies also existed in an Italian sample of traumatic brain-injured (TBI) patients. METHODS: A follow-up investigation was conducted on a sample of 16 subjects who had suffered severe TBI 5 to 10 years earlier. Based on series of neuropsychological tests, the patients' levels of remaining autonomous function were assessed. For each patient, a family reference person was identified with a semistructured interview to define how they dealt with the consequences of the trauma within the familial nucleus, what motivated their choices, the social outcome of the familial nucleus, the behavioral outcome, and the social- and work-related outcomes of the patient. The caregiver's current emotions were also analyzed. RESULTS: The study results confirmed that the family felt isolated from the social groups it used to belong to, and that few patients had fully achieved reinsertion into their social and/or work environments. Only 50% of subjects had returned to their job, most of them finding work at a lower level. The closest relatives reported experiencing a state of hardship. Many (70%) care-givers expressed by a steady feeling of worry but had devised effective compensatory strategies over the course of the study period to deal with depression. Such compensation appeared to be the result of the care-givers' awareness of the need to cope with situations where the patients very often depended on them. The hardship level the patient's familial nucleus experienced was not correlated with the patient's social- and work-related outcome, and elements of difficulty were evident among the relatives of patients who had returned to a productive life and those who were close to the family. The presence of severe cognitive and/or behavioral disability was a major factor in generating concern and worry about the future. In cases where the patient was severely disabled, the happiness level perceived by the patient's relatives, as expressed on a 5-point Likert scale, was 2.1, whilst in cases where disability was absent the level was 3.5. CONCLUSIONS: Our observations confirm evidence from previous studies that even many years after a severe trauma incident, a state of disability and/or hardship involving the entire familial nucleus persists. PMID- 16474281 TI - Critical comparison of nine different self-administered questionnaires for the evaluation of disability caused by low back pain. AB - AIM: The aim of this study was to critically compare 9 self-administered questionnaires designed to evaluate disability caused by back pain. METHODS: The questionnaires were analyzed considering and comparing their psychometric characteristics (reliability, validity, responsiveness, minimal clinically relevant difference), together with other practical and technical aspects (number of items, number and kind of domains, scaling of items, scoring, time to complete, validated translations, etc.). Data were obtained from scientific literature. RESULTS: Only 3 out of 9 analyzed instruments (i.e., the Oswestry Low Back Pain Disability Questionnaire, the Quebec Back Pain Disability Scale, and the Roland-Morris Low Back Pain Disability Questionnaire) appear fully validated from a psychometric standpoint. CONCLUSIONS: On the basis of psychometric evaluations as well as feasibility considerations, the authors suggest using either the Roland-Morris or Oswestry questionnaire as the best assessment of the level of disability caused by back pain. PMID- 16474282 TI - Shoulder-hand syndrome after stroke. A complex regional pain syndrome. AB - Complex regional pain syndrome (CRPS) types I and II are neuropathic pain disorders that develop as an exaggerated response to a traumatic lesion or nerve damage, that generally affects the extremities, or as the consequence of a distant process such as a stroke, spinal lesion or myocardial infarction. It rarely appears without an apparent cause. CRPS of upper limbs after stroke is frequently today called shoulder-hand syndrome (SHS). The onset and severity of SHS appears to be related with the aetiology of the stroke, the severity and recovery of motor deficit, spasticity and sensory disturbances. Another important aetiological factor is glenohumeral subluxation. The physiopathology of the disease is still not known. In CRPS, there is an exaggerated inflammatory response and some chemical mediators have been identified and are present in the inflammatory soup around the primary afferent fibres that, through different processes, can induce hyper-excitability of the afferent fibres (peripheral sensitization). It is hypothesized that a localized neurogenic inflammation is at the basis of oedema, vasodilation and hyperhidrosis that are present in the initial phases of CRPS. The repeated discharge of the C fibres causes an increased medullary excitability (central sensitization). Another important factor is the reorganisation of the central nervous system, and in particular this appears to affect the primary somatosensory cortex. The central role of the sympathetic nerve is presently in doubt. However, it is thought that a sub-group of CRPS patients exists in whom a predominant factor is the hyper-activity of the sympathetic nervous system, and that it responds positively to sympathetic block. Diagnosis is clinical and there are no specific tests, nor pathognomic symptoms to identify this disease with certainty. Diagnosis of CRPS after stroke appears more complex than in other pathological situations: the paretic upper arm frequently appears painful, oedematose, with altered heat and tactile sensations and slightly dystrophic skin within a non-use syndrome. Some investigations can aid differential diagnosis with other diseases. Treatment may be non pharmacological, pharmacological, with psychotherapy, regional anaesthesia, neuromodulation and sympathectomy. In any case there is little evidence that supports the efficacy of the interventions normally used to treat or prevent CRPS SHS. The key to effective treatment undoubtedly lies in a an expert multidisciplinary team that is co-ordinated and motivated and that treats the disorder with individualised therapy. PMID- 16474283 TI - Cerebral blood flow velocity during postural changes on tilt table in stroke patients. AB - AIM: The aim of the study is to investigate the correlation between orthostatic hypotension (OH), mean flow velocity (MFV) as measured by transcranial Doppler (TCD) in middle cerebral artery (MCA) bilaterally during tilt table test (TTT) and functional and neurological parameters in acute ischemic stroke patients, undergoing rehabilitation. METHODS: Thirteen patients after first ischemic stroke in the MCA territory and 13 healthy volunteers were examined. TTT was performed with elevating the subject from supine to 80 masculine standing position and back to supine within 10 min. Peripheral blood pressure was measured and monitoring of MFV in MCA of damaged and healthy hemisphere by TCD was performed during the TTT. Patients underwent the test few days after admission to rehabilitation department and were classified into 2 subgroups: those with decrease of systolic blood pressure of at least 20 mm/Hg during the test and those without the OH. RESULTS: Among patients without OH, MFV indexes were almost the same in damaged and healthy MCA and did not changed during the test. Patients with OH symptoms showed significant differences between blood flow velocities in 2 hemispheres in favor of nondamaged size. No association was found between the OH and the severity of functional and neurological status after stroke. CONCLUSIONS: These findings suggest that the appearance of OH after ischemic stroke may be associated with decreased blood flow velocity in damaged MCA at the beginning of rehabilitation treatment, but not with functional and neurological status of the patient. PMID- 16474284 TI - Effectiveness of functional magnetic versus electrical stimulation in women with urinary incontinence. AB - AIM: Urinary incontinence is one of the most common medical complaints in women. We here propose to evaluate and compare the effects of 2 conservative treatment modalities, functional electrical stimulation (FES) and functional magnetic stimulation (FMS). METHODS: We studied 22 female patients with urinary incontinence and divided them into 2 treatment groups (14 patients in the FES and 8 in the FMS group). The mean age of the patients in the FES group was 51.14+/ 11.9 and in the FMS group 42.25+/-6.9 years. Functional electrical stimulation was applied continuously at 10 Hz and 30-50 Hz in urge and stress urinary incontinence respectively. In mixed urinary incontinence stimulation was applied at 10 Hz for 15 min and at 50 Hz for 15 min. The treatment sessions were for 20 min, 3 times a week for 6-8 weeks (12 with mixed, 2 with stress incontinence). FMS was applied by a magnetic chair, twice weekly for 6 weeks (6 with mixed, 1 with stress urinary and 1 with urge urinary incontinence). The efficacy of the treatment was judged from patient impressions, records in urinary diaries, results of 1 h pad test, perineometry value and digital palpation score. RESULTS: The perineometry value, digital palpation score increased significantly during stimulation compared with prestimulation levels in both groups (P<0.05). For the pad test significant improvement was also noted in both groups (P<0.05). The urinary diaries and frequency of micturition were significantly more cured or improved in the FES group (P<0.05). However, reduction of the frequency of nocturnal micturition wasn't significant in either group (P>0.05). CONCLUSIONS: Both FES and FMS treatments were effective. FMS does not involve intravaginal stimulation and it is twice a week. Although FMS is not often used it is more cost effective than FES. In order to have exact knowledge of this issue; more research than has been done in a greater number of subjects is required. PMID- 16474285 TI - Correlation of back pain, compression fracture and quadriceps muscle strength with bone mineral density in renal insufficiency patients. AB - AIM: The purpose of this study was to evaluate bone mineral density (BMD), bone parameters, complete blood count, erythrocyte sedimentation rate, electrolyte values, back pain, compression fracture and quadriceps muscle strength in mild and moderate renal insufficiency patients. METHODS: Thirty-six female, 3 male, patients with the diagnosis of osteoporosis in addition to mild or moderate chronic renal insufficiency who were followed in Istanbul University, Cerrahpasa Faculty of Medicine, Physical Therapy and Rehabilitation Department, Osteoporosis and Nephrology Outpatient Clinics between March 2003 and March 2004, were included in the study. In the control group there were 17 female, 5 male patients with osteoporosis but without renal insufficiency. The inclusion criteria were to have osteoporosis, be aged between 40-70 with a creatinine clearance between 30 70 mL/min in the case group, >70 mL/min in the control group. The patients whose creatinine clearance was <30 mL/min, and whose BMD was normal or osteopenic even though creatinine clearance was >30 mL/min were excluded from the study. RESULTS: There was no significant difference with respect to back pain, compression fracture and quadriceps muscle strength between the 2 groups. The mean value of neck BMD, T and Z score were significantly lower in the case group (P<0.05). CONCLUSIONS: As a conclusion, PTH related bone disease had an important effect on BMD, although, the risk factors for osteoporosis were equally important. In various researches, the relationship between BMD values and history of fracture in renal osteodystrophy patients could not be shown. In our study, the history of prior fracture or fracture in the family were important risk factors for osteoporosis. Renal osteodystrophy patients had low BMD values in mild and moderate stages of the disease. Prior fracture as well as osteoporotic fractures in the family should be part of a comprehensive evaluation of the patient. PMID- 16474286 TI - Depression and psychosocial factors in Turkish women with fibromyalgia syndrome. AB - AIM: The aim of this study was to evaluate the demographic qualifications, frequency of depression, degree of pain, and the correlations between these factors in Turkish women with fibromyalgia syndrome (FS). METHODS: Fifty-three women with FS and 54 healthy women were included in the study. The visual analogue scale (VAS) was applied to evaluate the degree of pain and Beck depression inventory (BDI) for depression. RESULTS: Mean age of the FS and the healthy women groups was 42.6+/-9.6 (21-63) and 39.4+/-13.2 (24-60) years respectively; which were statistically similar (P>0.05). Of the patients, 69.8% were married, and the mean years of education was 8.7+/-4.8 years. Marriage among relatives was found in 18.4% of the patients. The mean VAS score of the patients was 7.2+/-1.7 cm with the mean duration of pain 5+/-4.6 years. Mean BDI scores of FS patients and the healthy group were 15.7+/-8.7 and 10.2+/-5.5 respectively; the difference between the 2 groups was statistically significant (P<0.05). According to the BDI scores, 90% of FS patients were classified as depressed; among them, 50% had minor, 38% moderate, and 2% severe depression. FS patients who were married to a relative had higher scores of BDI (r=0.414, P=0.013). There was negative correlation between BDI score and patients' total year of education (r=-0.295, P=0.037); and the husband's education level (r=-0.367, P=0.030). According to BDI, the c2 test revealed significant depression in patients with sleep-disorders (P=0.009). CONCLUSIONS: We found a significant degree of depression in Turkish female FS patients. This situation is found to be correlated with the education level of both patient and husband; marital status, and sleep-disorder; it is suggested that these factors should be taken into consideration in the diagnosis, treatment and follow-up of FS patients. PMID- 16474287 TI - SIMFER Rehabilitation treatment guidelines in postmenopausal and senile osteoporosis. PMID- 16474288 TI - [Modern methods of ocular hemodynamics assessment in surgical correction of visual ischemic disturbances]. AB - The paper describes management results of 56 patients with occlusive and stenotic atherosclerotic lesions of carotid arteries, complicated by different forms of ocular ischemia. Complex assessment (duplex scanning, color Doppler mapping, transcranial Doppler) before and a year after carotid endarterectomy revealed a significant improvement in objective parameters of ocular hemodynamics: registration of antegrade blood flow in ophthalmic artery, the rise of maximal systolic blood flow velocity and decrease of resistance index in ophthalmic, central retinal and posterior ciliary arteries. PMID- 16474289 TI - [Changes in hemostasis at aortic surgery under conditions of extracorporeal circulation]. AB - This paper describes hemostatic changes in patients who had undergone aortic surgery under conditions of extracorporeal circulation. The changes in the hemostatic system are marked by the thrombophilic condition made up for activation ox blood fibrinolytic activity in the preoperative period. The basic amount of blood loss falls within the first 6 hours after operation. Prolonged hypothermic extracorporeal circulation and the high intraoperative blood loss (over 35 ml/kg bw) lead to an appreciable decrease in antithrombin III and protein C activity which results in activation of disseminated intravascular blood coagulation in the early postoperative period and ineffectiveness of heparin therapy. Secondary hyperfibrinolysis together with combined thrombocytopenia and platelet dysfunction are the basic causes of higher than usual bleeding in patients after aortic surgery under conditions of extracorporeal circulation. PMID- 16474290 TI - Strategy and tactics of instrumental examination of patients with thoracic and thoracoabdominal aortic aneurysm. AB - The paper gives a detailed characterization and describes the importance of instrumental methods for examination of patients in the diagnosis of thoracic and thoracoabdominal aortic aneurysms. Provides a differentiated protocol of the examination of patients as dependent on the clinical course, disease stage and nemodynamic stability of the patient's status. Delineates the tactics of the examination of patients with acute and chronic dissection of the proximal and distal segments of the aorta as dependent on hemodynamic stability. Emphasis is laid on the key role of computed tomography and magnetic resonance tomography in the diagnosis of aortic aneurysms. PMID- 16474291 TI - Carotid angioplasty stenting under protection: techniques, results, limitations (part 1). PMID- 16474292 TI - Impact of the volume of aneurysmal contents on intraaneurysmal pressure after endovascular grafting (experimental studies). AB - With the purpose of clarifying the nature and outlining certain constituents of such an undesirable condition designated as "endotension", that emerges after transluminal grafting of the aneurysmally changed aorta, a basic theoretical model was worked out for explanation of "endotension". Also, there was designed and constructed in vitro an original experimental model using which the authors carried out a study into the relationship between the pressure in the aneurysm after its complete exclusion from the blood flow by an endovascular graft and the volume of the aneurysmal contents. Some factors described in the literature as influencing the pressure level in the aneurysm were at the given stage purposefully excluded. The volume of the "aneurysm" reproduced in our model was equal to 675 ml; each of five vascular grafts implanted into the aneurysm measured 110 mm in length. The pulsating liquid flow was reproduced in the system using a serial appliance for extracorporeal circulation. The liquid was aspirated from the aneurysm by means of a syringe with concurrent pressure guidance in the sac. To start pressure lowering, it was necessary to evacuate 0.4-1.6 ml of the liquid. The real clinical situations were considered from the standpoint of our results. PMID- 16474293 TI - [Derivatives of semi-synthetic diosmine in the treatment of patients with chronic venous insufficiency. The results of a prospective study--using phlebodia 600]. AB - This paper describes the results of an open, prospective, non-comparable study of the efficacy and tolerance of the drug phlebodia 600 mg in patients with lower limb CVI. The study accrued 32 patients aged 21-70 years suffering from lower limb varicosity coupled with class I-IV CVI according to the CMP. Of the objective criteria, there were made measurements of the malleolar volume. The grade of subjective symptoms was recorded in terms of the clinical scale for evaluating the severity of the VCSS disease, which is a subdivision of the CEAP classification. The quality of the patients' life was examined before the patient was included into the study and after its completion using a standard CIVIQ questionnaire. The positive time course of changes was marked in all the constituents of the integral indicator of the patient's quality of life. The most significant change in the patients' quality of life was identified in patients in clinical class III according to the CEAP. The authors analyze the reports concerning the use of the above-indicated drug in European phlebological practice. PMID- 16474294 TI - Radionuclide assessment of the treatment by compression of patients with chronic venous insufficiency. AB - A study was made of the efficacy of the compression medical tricot in the treatment of chronic venous insufficiency (CVI). To evaluate the treatment by compression, the authors, in addition to the clinical examination, used radionuclide techniques such as examination of the regional blood volumes and phleboscintigraphy. The results of the given work have shown that the compression products under test are effective agents for the treatment of CVI by compression which improves venous return and the patients' quality of life. The data obtained point out that in addition to the lowering of blood filling, the medical tricot influences the interstitial element of the edematous syndrome, leading to the elimination of the latter one. The use of elastic bandages creates the known difficulties (the necessity of the patient training in bandaging, complexity of creating an adequate bandage, quick depreciation). In view of this fact the use of the compression tricot which possesses the fixed grade of compression for a concrete form and stage of CVI is preferable in the therapy of chronic venous diseases. PMID- 16474295 TI - [The use of Ginkor Fort in the program of multimodality rehabilitation treatment of CYI patients with edematous syndrome]. AB - Edematous syndrome is one of the most frequently occurring manifestations of chronic venous insufficiency (CVI). This paper analyzes an experience with conservative treatment of CVI patients with edematous syndrome. A complex of rehabilitation measures was used in addition to therapy by the phleboprotector Ginkor Fort. Based on the study of microcirculatory changes, malleolar volume, the thickness of subcutaneous fat as well as on subjective sensations and the quality of life in 117 patients with the edematous forms of CVI the authors have demonstrated the high efficacy of combined pharmacotherapy by Ginkor Fort and rehabilitation treatment in the given patient group. PMID- 16474296 TI - [Pulmonary artery thromboembolism in patients with trauma]. AB - The study accrued 257 patients with mechanical traumas of varying severity and sites, complicated by post-traumatic phiebothrombosis. In 87 (33.85%) patients, traumatic disease was aggravated by pulmonary artery thromboembolism (PTE). In the course of the study we evaluated risk factors of PTE related to the patient's condition and trauma. It has been established as a result that of 14 recognized risk factors of PTB determined by the patient's condition, only six influenced this disease outcome, namely: age over 40 years, a history of PTE or deep venous thrombosis (DVT), overweight, circulatory and respiratory insufficiency, bed regime. Among risk factors of PTE linked with trauma, the following factors had a significant impact on the emergence of PTE: treatment by means of skeletal traction, the site of trauma in the lower extremity (femoral or tibial bone), the site of previous thrombosis in the deep veins of the leg and thigh as well as floating thrombus. PMID- 16474297 TI - Intraoperative ruptures of thoracic aortic aneurysms. AB - Intraoperative rupture of thoracic aortic aneurysm is a severe complication which may have an adverse effect on the outcome of operation. Among all intraoperative aneurysmal ruptures the most difficult and uncontrolled are ruptures of aortic aneurysms in the presence of dissection and of false aortic aneurysm during repeat interventions in this area. At intraoperative aortic rupture the priority task of the surgical team consists in urgent attainment of maximal temporary hemostasis and an adequate replenishment of blood loss. Presented herein are two most demonstrative clinical cases of intraoperative aortic ruptures: of the ascending aorta during cannulation of the right atrium and of false aneurysm of the aortic isthmus, which developed after aortic isthmoplasty by dacron patch for coarctation where the posterolateral wall of aneurysm was visceral pleura of the left lung. The authors provide a detailed description of the treatment policy for the given condition. Review the reported data pertaining to the problem under consideration. PMID- 16474298 TI - Revascularization in distal lesions of upper extremity arteries. AB - A study was made of the course of ischemia and hemodynamic disorders in 53 patients with occlusion of the distal arterial system of the upper extremities. Circulation was examined by ultrasound Doppler, tetrapolar rheology, transcutaneous oxymetry and angiography. Critical ischemia was discovered in 58.5% of patients. In occlusion of one of the forearm arteries, extremity circulation was made for. Circulatory decompensation was recorded in occlusion of both forearm arteries with intact distal bed and non-functioning palmar arches of the hand as well as in lesions of hand and finger arteries. All 53 patients were operated on, 66 operations were accomplished using magnifying optic appliances. Provided the distal bed was well preserved, use was made of direct revascularization techniques (36) whereas non-standard methods were employed in the event of its obliteration: arterialization of the outlets of the subcutaneous veins of the hand and autotransplantation of the greater omentum (30). Beneficial results were obtained in 91.5% and unsatisfactory in 8.5% of patients. The lethality accounted for 1.2% while the incidence of amputations was quoted as 5.7%. Good results offer the period as long as 5 years were well preserved in 87.6% of patients. Our experience indicates the efficacy of the use of revascularization techniques for the treatment of different versions of occlusion of the distal segments of upper extremity arteries. PMID- 16474299 TI - [Extraanatomic reconstruction of the right renal artery]. AB - This paper reviews two clinical cases of a rare type of extraanatomic revascularization of the right kidney by means of establishing a renogastroduodenal arterial anastomosis. The clinical cases presented herein demonstrate an adequacy of the given extraanatomic reconstruction of the renal artery. The indications for such operation are compelled and should be determined strictly individually in terms of each concrete case. PMID- 16474300 TI - Carotid artery dissection (literature review and a case report). AB - The paper presents a literature review on the internal carotid artery (ICA) dissection and an original case report. The dissected ICA segment was resected and grafted. PMID- 16474301 TI - [The use of pentoxyphylline in the treatment of patients with chronic obliterative diseases of lower limb arteries]. AB - The review concerns the assessment of the efficacy of pentoxyphylline which is most popular today in the treatment of patients with chronic arterial lower limb insufficiency. The new data are presented on its action mode as are multiple examples of the use of pentoxyphylline in the treatment of patients with peripheral angiopathies. PMID- 16474302 TI - [In-hospital and remote results of the use of sirolimus coated coronary stent CYPHER]. AB - Stents were implanted to 554 previously non-revascularized patients in 703 injured coronary segments; 32% of stented stenoses were complicated; 23% of patients received 2-4 stents. Immediate success was 100% with no cases of acute stent thrombosis. Rate of in-hospital subacute thrombosis and non-Q-wave myocardial infarction was 0.2%. Survival without restenosis, angina, myocardial infarction and repeat revascularization during follow up (mean duration 257+/-107 days) was 94%, restenosis rate -- 5.6%. Six patients with in-stent restenosis were successfully revascularized. All 108 patients followed for 27 months were alive by the end of this term. PMID- 16474303 TI - [Antibodies to Chlamydia pneumoniae in blood serum of patients with ischemic heart disease and presence of complicated lesions in coronary arteries]. AB - Levels of IgM, IgG and IgA antibodies to Chlamydia pneumoniae were measured in 107 patients (age 33-75 years) with documented coronary atherosclerosis and 39 subjects with intact coronary arteries. Rates of seropositivity to C. pneumoniae were 77.6 and 25.6% in patients and "healthy" subjects, respectively (p<0.05). Seropositive (n=83) compared with seronegative (n=24) patients had higher prevalence of complicated lesions (p<0.05). PMID- 16474304 TI - [Dynamics of prevalence of ischemic heart disease and risk of cardiovascular death in open population of Tiumen]. AB - Prevalence of ischemic heart disease (IHD) has been repeatedly studied in 3 sex and age stratified representative samples of Tyumen citizens. Prevalence of IHD in Tyumen open population is 11.7 among men and 13.2% among women. Prevalence of definite IHD is 7.1 and 8.4%, respectively. Studies conducted with 5 years interval demonstrated rise of prevalence of IHD defined according to soft criteria among women at the account of its probable forms. Relative risk of cardiovascular death rose in middle aged and elderly men and in elderly women. PMID- 16474305 TI - [Prognostic value of C-reactive protein and neopterin in autologous bone marrow cells transplantation]. AB - Relationship between levels of C-reactive protein (CRP) and neopterin and changes of left ventricular function was studied in 23 patients with heart failure (mean age 48.5+/-11.5 years) subjected to transplantation of autologous bone marrow cells (TABMC). Cells were administered intracoronary irrespective of coronary revascularization (in 11 patients) or during angioplasty (in 5 patients), intracoronary or intra-cardially during aorto coronary bypass surgery (in 7 patients). Reverse correlation was observed between left ventricular ejection fraction (EF) and stroke volume (SV) and blood plasma levels of CRP and neopterin. Efficacy of functional regeneration of the myocardium assessed 1 month after TABMC was related to blood plasma levels of CRP and neopterin before treatment. In all patients with normal initial levels of markers of inflammation EF and SV rose. Among patients with initially elevated levels of CRP and neopterin more positive changes of EF and SV occurred in those subjects in whom lowering of inflammation markers took place during 1 month after TABMC. PMID- 16474306 TI - [Effect of 6-month therapy with simvastatin on lipid transport function of the blood and the state of endothelium in patients with diabetes and hypertension]. AB - The state of lipid transport function of the blood, blood contents of stable nitrogen metabolites, and proinflammatory cytokines (TNF-a and IL-1b) during therapy with simvastatin were studied in 29 patients receiving combination antihypertensive therapy with angiotensin converting enzyme inhibitors (ACEI) and verapamil. Lipid lowering action of simvastatin was realized just in 1 month of treatment and remained sustained for half a year (average duration). 6 months after addition of simvastatin to antihypertensive therapy substantial (58.4%, p=0.044) rise of plasma content of stable nitrogen metabolites took place. At the same time therapy with metoprolol in a similar group of patients exerted no considerable effect on blood plasma concentration of nitrate and nitrite anions. Lowering of median values of TNF-alpha from 20.13 (12.67-52.80) to 11.34 (3.31 31.29) pg/ml (p<0.0038) was also noted at the background of combination antihypertensive therapy. This happened without distinct affair with degree of lipid lowering action of simvastatin. The results of the study document positive effect of half year treatment of patients with concomitant hypertension and diabetes with simvastatin (10-20 mg/day) in combination with ACEI and verapamil on metabolism of nitric oxide and plasma content of TNF-alpha which realizes independently from degree of hypolipidemic action of simvastatin. PMID- 16474307 TI - [Noninvasive diagnosis of the state of lower extremities arterial vascular bed in patients with diabetes of type 2]. AB - Complex noninvasive investigation of arterial vascular bed and microcirculation of lower extremities was carried out in 87 patients with diabetes and 47 patients with atherosclerosis obliterans of lower extremities without diabetes. Color duplex scanning allowed to detect occlusive-stenotic and nonocclusive changes of arterial vascular bed in patients with type 2 diabetes. Calcinated atherosclerotic plaques in patients with type 2 diabetes more often localize in superficial femoral, popliteal arteries and arteries of the calf. Such plaques are twice as frequent among patients with atherosclerosis obliterans with diabetes compared with those without diabetes. Laser Doppler flowmetry with test of reactive postocclusion hyperemia and positional test in patients with atherosclerosis obliterans and type 2 diabetes allows to reveal more pronounced disturbances of microcirculation compared with patients without diabetes. In patients with type 2 diabetes with hemodynamically insignificant stenoses of arteries of lower extremities laser doppler flowmetry and transcutaneous measurement of oxygen tension in skin of a foot reveal disturbances of microcirculation. PMID- 16474308 TI - [Prognostic significance of parameters of heart rate variability as a risk factor of development of hypertension]. AB - Risk factors and parameters of heart rate variability during active orthoclinostatic test in subjects with normal blood pressure were assessed in a 5 year cohort study on 519 individuals. Logistic stepwise regression revealed most informative factors associated with development of hypertension: sex (p=0.029), body mass index (p=0.005), salt sensitivity threshold (p=0.048), dynamics of heart rate during exercise test (p=0.035). Addition of parameters of heart rate variability (dynamics of standard deviation of R-R intervals and of low frequency component of spectrum during active orthoclinostatic test) increased accuracy of prognosis. With the use of constructed models risk profiles were calculated. Thus, parameters of heart rate variability allow to improve prognostic value of a model. PMID- 16474309 TI - [Comparative efficacy and safety of contemporary Angiotensin converting enzyme inhibitors moexipril and spirapril in women with postmenopausal metabolic syndrome]. AB - Moexipril (7.4-15 mg/day) was given to 34, spirapril (3-6 mg/day) -- to 18 postmenopausal women with hypertension and metabolic syndrome for 16 weeks. Hydrochlorthiazide was added when therapy was not sufficiently effective. Both angiotensin converting enzyme inhibitors had similar hypotensive activity: blood pressure normalized in 71 and 61% of moexipril and spirapril treated women, respectively. Both drugs promoted normalization of metabolism of lipid (lowering of levels of cholesterol, atherogenic lipoproteins and triglycerides) and carbohydrates (lowering of hyperinsulinemia). Patients with postmenopausal metabolic syndrome had elevation of leptin level up to 27.5+/-5.5 pg/ml. Moexipril and spirapril caused lowering of elevated levels of leptin. These drugs did not affect levels of sex hormones. They exerted vasoprotective (normalization of endothelium dependent and independent vasodilatation) and nephroprotective (attenuation and normalization of microalbuminuria) effects. Thus spirapril and moexipril are effective in treatment of hypertension in patients with postmenopausal metabolic syndrome. PMID- 16474310 TI - [Changes of cerebral perfusion and cognitive function in patients with essential hypertension at the background of hypotensive therapy]. PMID- 16474311 TI - [Brain natriuretic peptide as marker and factor of prognosis in chronic heart failure]. AB - The role of natriuretic peptides in pathogenesis of chronic heart failure (CHF) is reviewed. Main attention is given to significance of measurement of brain natriuretic peptide in the following clinical situations related to CHF: diagnosis of CHF, screening of patients with symptomless left ventricular dysfunction, differential diagnosis of edematous syndrome, indications to therapy and monitoring of its effectiveness, assessment of long term prognosis of patients with CHF. Possibilities of curative use of natriuretic peptides and inhibitors of vasopeptidases are also discussed. PMID- 16474312 TI - [Chronic fatigue syndrome in cardiology neurohumoral changes]. AB - Chronic fatigue markedly worsens quality of life of cardiological patients. Chronic fatigue and chronic fatigue syndrome are neuro-immuno-endocrine disorders which manifest as moderate and severe even invalidizing fatigue with psychosomatic symptoms. External and internal stress such as psychological stress, stress after major surgery and trauma, depressive states, inadequate physical exercise, chronic heart failure, chronic viral infection, oncologic diseases, -- can promote development of chronic fatigue. Immune and hypothalamic pituitary-adrenal (HPA) axis abnormalities were found to be associated with this condition. Measurement of plasma cortisol concentration is used as basic characteristic of HPA axis function. Measures aimed at detection of chronic fatigue in cardiological patients and its appropriate management should supplement programs of integrated rehabilitation in order to improve quality of life and facilitate return to work. PMID- 16474313 TI - [Differentiated use of nitrates in ischemic heart disease]. AB - Clinical aspects of the use of organic nitrates in the treatment of patients with ischemic heart disease (IHD) are considered. Efficacy of organic nitrates in various forms of IHD as well as in patients with concomitant diseases is discussed. Special stress is made on clinical aspects of application of nitrates in treatment of IHD. Possibilities and safety of contemporary nitrates in patients with various forms of IHD are discussed. PMID- 16474314 TI - [Invasive methods of treatment and prevention of ventricular tachyarrhythmias after myocardial infarction]. PMID- 16474315 TI - [Marfan syndrome]. AB - Marfan syndrome is a hereditary disease of connective tissue with predominant involvement of cardiovascular system, skeleton and eyes. Timely diagnosis and adequate management can substantially decrease mortality in this condition. The review summarizes data of clinical observations and studies of etiology, pathogenesis, diagnosis and treatment of Marfan syndrome. PMID- 16474316 TI - An APRIL to remember: novel TNF ligands as therapeutic targets. AB - Since their discovery in 1998, the two TNF family members APRIL and BLyS/BAFF have received increasing attention. In addition to regulating normal B-cell development and immune responses, these molecules might be crucial in a diverse set of diseases, including autoimmunity and cancer. Although more has been published about the general biology of BLyS/BAFF than that of APRIL, many recent articles have described novel APRIL biology. Here we focus on APRIL, exploring its normal and pathological functions, and comparing the therapeutic molecules currently under development that target BLyS/BAFF alone, or APRIL and BLyS/BAFF together. PMID- 16474317 TI - Of bright college days or expedient solutions? PMID- 16474318 TI - Disturbing conclusions. PMID- 16474319 TI - Lack of evidence. PMID- 16474321 TI - No more infected sockets? PMID- 16474320 TI - Commercialism in marketing. PMID- 16474322 TI - Praise for ozone. PMID- 16474325 TI - Edinburgh degrees. PMID- 16474323 TI - Practising with pacemakers. PMID- 16474326 TI - Bring back compassion. PMID- 16474327 TI - Saliva test. PMID- 16474328 TI - New dental school to be launched in the South West. PMID- 16474330 TI - Mogjan Azari struck off. PMID- 16474342 TI - Contract will fail patients, BDA warns Minister. PMID- 16474349 TI - Cardiff dental researchers tackle childhood tooth decay. PMID- 16474350 TI - Liquorice root may help fight tooth decay. PMID- 16474351 TI - Aesthetic changes with four anterior units. AB - Cosmetic dentistry has evolved with the advent of more robust porcelain materials and ever-stronger bonding agents. This series of three articles aims to provide a practical overview of what is now possible both functionally and cosmetically from the preparation of a small number of teeth, through a whole smile, to full mouth rehabilitation. A complete diagnosis is the starting point to planning any cosmetic or functional changes. Guidance is given on the techniques used but adequate training must be considered essential before embarking upon modification in occlusal schemes or even minor adjustments in smile design. Conservative use of porcelain laminate veneers can result in significant aesthetic improvements to a patient's smile and it is not always necessary to prepare all the teeth visible in a smile. PMID- 16474352 TI - Concrescence of a mandibular third molar and a supernumerary fourth molar: report of a rare case. AB - Concrescence represents a rare developmental anomaly in which two fully formed teeth are joined along the root surfaces by cementum. Maxillary molars are the teeth most frequently involved, especially a third molar and a supernumerary tooth. Very few cases have been reported about the concrescence of a third molar and a supernumerary tooth. According to our current knowledge, this case report is the first in the literature in which concrescence is observed between a third molar and a supernumerary fourth molar in the mandible. PMID- 16474360 TI - Indirect retention. PMID- 16474361 TI - Potential adverse effects. PMID- 16474362 TI - Criteria for CPD. PMID- 16474363 TI - Determining "need" for a Removable Partial Denture: a qualitative study of factors that influence dentist provision and patient use. AB - OBJECTIVE: To identify factors that influence Removable Partial Denture (RPD) provision, and patient use of RPDs in the UK. DESIGN: Exploratory qualitative interview study. SUBJECTS AND METHODS: SUBJECTS: There were two sample groups. A purposive sample of 16 male and female dentists was categorised in terms of level of RPD provision, experience, and practice characteristics. A purposive sample of 17 male and female partially dentate patients was categorised in terms of RPD use and demographic characteristics. DATA COLLECTION: Semi-structured in-depth interviews. RESULTS: For dentists, RPD provision was indicated by patient demand and physical function of the remaining teeth, but was mediated by NHS fee structures and professional satisfaction. For patients, RPD use was influenced by the trade-off between improved appearance and the unpalatable presence of an RPD in their mouth. The location of the gap(s) was important, but other issues were relevant such as ability to "manage" without the RPD. CONCLUSION: When defining "need" for an RPD, dentists focused on physical function of the teeth whereas patients focused on social meanings of the mouth. These differing priorities may improve understandings of patient non-compliance in RPD use. Further research on the relationship between denture use and social identity could be beneficial. PMID- 16474364 TI - Perceptions of how the Internet has impacted on dentistry. AB - OBJECTIVES: This study sought to identify how patient information on the Internet has influenced the delivery of oral care and the use practitioners themselves make of the Internet. METHODOLOGY: Data were collected via a self completed 18 item postal questionnaire, sent to a random sample of 620 dentists on the GDC register in Wales. RESULTS: In total 457 (74%) of the questionnaires were returned. One half of all practitioners stated that patients had asked them about material of relevance to dentistry obtained from the Internet, although in the majority of cases this happened less frequently than once a month. The most common topics enquired after were cosmetic procedures, dental amalgam and implants. A minority of dentists, 47 (11%) viewed the Internet as a threat to the dentist-patient relationship. However, 169 (39%) agreed information gained from the Internet had led to patients demanding inappropriate care or more complex treatment (135/31%). Having to take time to discuss Internet material was viewed as a burden by 93 practitioners. The potential of the Internet to widen inequalities in access to oral health information was agreed upon by one third of respondents. Concerns over the quality and reliability of Internet derived information, together with a lack of knowledge of appropriate sites, prevented dentists using the Internet as an oral health education resource. However, only 18.7% claimed never to use the Internet for their own Continuing Professional Development. Access for CPD purposes was mainly from home. Of those working in general practice, 54 had their own practice website, and a further 103 said this was something they were considering. Email was used to communicate with patients by 42 dentists, mainly to make appointments. CONCLUSIONS: Views on the impact of the Internet were generally positive, but there is a long way to go before its full potential is realised. PMID- 16474365 TI - Awareness of mouth cancer in Great Britain. AB - OBJECTIVE: There are more than 2,000 new cases of mouth cancer each year in Britain. Early detection is important yet little is known about population awareness of this disease and ability to recognise early signs, particularly among high risk groups. This study aimed to address this issue. METHODS: Data were collected by means of household survey. A total of 3,384 adults were questioned using a national probability sample. Respondents provided information on demographic characteristics, smoking status, and frequency of alcohol use. They were asked whether they had heard of mouth cancer. Their knowledge of early signs and risk factors was assessed. RESULTS: Whereas 95.6% of respondents said they had heard of mouth cancer, their awareness of early signs was low; for example, only 33.8% recognised that white patches in the mouth were a sign. The large majority understood that smoking and chewing tobacco were risk factors (84.7% and 80.1% respectively) but only 19.4% recognised alcohol use as a risk factor. In multiple logistic regression analyses controlling for relevant demographic factors, smokers and those with more frequent alcohol consumption were less likely to recognise early signs. CONCLUSIONS: Awareness of early signs of mouth cancer is low and lower in people who as a result of their behaviour are at higher risk. There is a need to raise awareness in those at most risk. PMID- 16474375 TI - Early-onset gastric cancers have a different molecular expression profile than conventional gastric cancers. AB - Many studies examine the molecular genetics of gastric cancer, but few look at young patients in particular and there is no comparison of molecular expression between early-onset gastric cancer (< or = 45 years old) and conventional gastric cancers. Expression of cycloxygenase-2 (COX-2) is elevated in gastric adenocarcinomas compared to non-neoplastic mucosa, and in light of studies showing reduced risk of gastric cancer in nonsteroidal anti-inflammatory drug users, we have chosen to investigate the expression of COX-2 and related molecules in 113 early-onset gastric cancers and compare it with 91 conventional gastric cancers, using tissue microarrays. These markers include molecules known to be important in conventional gastric carcinogenesis, such as E-Cadherin, p53, COX-2, Trefoil Factor-1 (TFF1), beta-catenin, p16 and c-myc; as well as molecules not yet described as being important in gastric cancer, such as the transcription factor c-jun, the COX-2 mRNA stabilizer HuR, and C/EBP-beta, a transcription factor for COX-2. All markers showed a statistically significant difference between early-onset gastric cancers and conventional gastric cancers, using a chi2 test. In particular, early-onset gastric cancers displayed a COX-2 Low, TFF1 expressing phenotype, whereas COX-2 overexpression and loss of TFF1 was found in conventional cancers, and this difference between early-onset gastric cancers and conventional cancers remained statistically significant when adjusted for location and histology (P<0.0001 and P = 0.002 respectively). We found that COX-2 overexpression correlates significantly with loss of TFF1 (P = 0.001), overexpression of C/EBP-beta (P<0.001) and cytoplasmic HuR (P = 0.016). COX-2 was significantly associated with p53 positivity (P = 0.003). Abnormalities in E Cadherin correlated significantly with diffuse phenotype, whereas high expression of COX-2, loss of TFF1 and overexpression of C/EBP-beta correlated with the intestinal phenotype. Our results provide further evidence that early-onset gastric cancer exhibits a distinctive expression profile that may have practical implications. PMID- 16474376 TI - Study of phospho-beta-catenin subcellular distribution in invasive breast carcinomas in relation to their phenotype and the clinical outcome. AB - Beta-catenin has a crucial role in cell-cell adhesion as well as a signaling role as a member of the Wnt pathway. The aim of this study was to examine the clinicopathological and prognostic value of phosphorylated beta-catenin, as well as its relation to the tumors' phenotype, in breast cancer. Immunohistochemistry was applied on 141 paraffin-embedded breast tissue specimens for the detection of phospho-beta-catenin, ER, PR, c-erbB-2, p53, Ki-67, bcl-2, uPAR and TIMP-1. For each case, a phospho-beta-catenin index was determined by image analysis. Phospho beta-catenin staining was detected in the cytoplasm and the nucleus of the malignant cells. Cytoplasmic phospho-beta-catenin was statistically higher in carcinomas of smaller tumor size (P = 0.030), lower stage (P = 0.026), decreased Ki-67 and high c-erbB-2 immunoreactivity (P = 0.052 and P = 0.037, respectively). Nuclear phospho-beta-catenin showed a parallel correlation with ER and ERbeta (P = 0.022 and P = 0.043, respectively), bcl-2 (P = 0.042), uPAR in cancer cells (P = 0.041) and TIMP-1, although the correlation was borderline (P = 0.066). Cytoplasmic phospho-beta-catenin was found to be independently correlated with prolonged disease-free and overall survival (P = 0.046 and P = 0.002, respectively), whereas nuclear localization was correlated with a shortened overall survival (P = 0.046). In conclusion, phospho-beta-catenin may have a different involvement in invasive breast carcinomas, according to its subcellular distribution. Nuclear localization seems to be related to an aggressive tumor phenotype, negatively affecting patients' overall survival, whereas cytoplasmic localization is associated with a favorable tumor phenotype and a longer disease free and overall survival. PMID- 16474377 TI - Clinicopathological significance of loss of heterozygosity in intestinal- and solid-type gastric carcinomas: a comprehensive study using the crypt isolation technique. AB - The clinicopathological significance of loss of heterozygosity (LOH) in gastric carcinoma remains poorly understood. We and other researchers have previously demonstrated that LOH is fairly common in intestinal- and solid-type gastric carcinomas, but rare in diffuse-type tumors. In this study, we investigated the relationship between clinicopathological variables and LOH status in intestinal- and solid-type gastric carcinomas. The crypt isolation technique was utilized to analyze LOH at 1p36, 3p14, 4p15, 5q21-22, 8p11-12, 9p21, 13q22, 17p13.1 18q21 and 22q13.31 in 113 intestinal- and solid-type gastric carcinomas using a polymerase chain reaction assay. Immunostaining with D2-40 and Elastica van Gieson staining were used to detect lymphatic invasion and vessel invasion, respectively. High LOH rates (49-71%) were observed in all chromosomal regions tested. 1p36 loss was significantly associated with advanced tumors and lymph node metastasis. 8p11-12 loss was significantly associated with lymph node metastasis, lymphatic invasion, and vessel invasion. 17p13.1 (TP53) loss was significantly associated with vessel invasion. 22q13.31 loss was significantly associated with advanced tumors, lymph node metastasis, lymphatic invasion, vessel invasion and late TNM stage. No significant associations were observed between LOH at other chromosomal regions and aggressive behaviors. In addition, significantly higher LOH rates at 1p36, 9p21, 18q21 and 22q13.31 were observed in cardiac tumors compared with noncardiac tumors. These results suggest that in intestinal- and solid-type gastric carcinomas, LOH on 3p14, 4p15, 5q21-22, 9p21, 13q22 and 18q21 is associated with carcinogenesis, while LOH on 1p36, 8p11-12, 17p31.1 and 22q13.31 is associated with tumor progression. PMID- 16474378 TI - High-grade urothelial carcinoma of the renal pelvis: clinicopathologic study of 108 cases with emphasis on unusual morphologic variants. AB - A clinicopathologic study of 108 cases of high-grade urothelial carcinomas of the renal pelvis is presented. Of the 108 tumors, 44 (40%) showed unusual morphologic features, including micropapillary areas (four cases), lymphoepithelioma-like carcinoma (two cases), sarcomatoid carcinoma (eight cases, including pseudoangiosarcomatous type), squamous differentiation and squamous cell carcinoma (15 cases), clear cells (two cases), glandular differentiation (two cases), rhabdoid, signet-ring or plasmacytoid cells (four cases), pseudosarcomatous stromal changes (four cases) and intratubular extension into the renal pelvis (three cases). Pathological staging was available in 62 patients; of these, 46 cases (74%) were in high stage (pT2-pT4) and 16 (26%) were in low stage (pTis, pTa, pT1). Clinical follow-up ranging from 1 to 256 months (median: 50 months) was available in 42 patients; of these, 26 (61%) died of tumor with a median survival of 31 months. The patients who did not die of their tumors showed only minimal or focal infiltration of the renal parenchyma by urothelial carcinoma, whereas those who died of their tumors showed massive infiltration of the kidney by the tumor. High-grade urothelial carcinomas of the renal pelvis can show a broad spectrum of histologic features similar to those seen in the urinary bladder. Our results support the finding that, unlike urothelial carcinomas of the bladder, the majority of primary urothelial carcinomas of the renal pelvis are of high histologic grade and present in advanced stages. Our study further highlights the fact that, in the renal pelvis, urothelial carcinomas show a tendency to frequently display unusual morphologic features and metaplastic phenomena. The importance of recognizing these morphologic variants of urothelial carcinoma in the renal pelvis is to avoid confusion with other conditions. The possibility of a high-grade urothelial carcinoma should always be considered in the evaluation of a tumor displaying unusual morphologic features in the renal pelvis, and attention to proper sampling as well as the use of immunohistochemical stains will be of importance to arrive at the correct diagnosis. PMID- 16474379 TI - Association of E-cadherin, matrix metalloproteinases, and tissue inhibitors of metalloproteinases with the progression and metastasis of hepatocellular carcinoma. AB - Molecular markers can provide additional information to traditional histomorphological evaluation for the assessment of tumor progression and predicting the likelihood of invasion and metastasis in various types of malignancies. We studied the association of E-cadherin, matrix metalloproteinases (MMPs), and tissue inhibitors of metalloproteinase with the progression and metastasis of hepatocellular carcinoma. Tissue microarray including six normal livers, 14 cirrhotic livers, 39 macroregenerative nodules, 16 dysplastic nodules, 22 grade I hepatocellular carcinomas, 43 grade II hepatocellular carcinomas, seven grade III hepatocellular carcinomas, and 10 metastatic hepatocellular carcinomas were stained immunohistochemically with antibodies against MMPs -1, 2, -3, -7, -9, tissue inhibitors of metalloproteinase-1, -2, -3, and E-cadherin. The intensities of staining were scored manually by two pathologists and verified by the Chromavision Automated Cellular Imaging System. Compared with normal liver, cirrhotic liver had significantly lower E-cadherin and tissue inhibitors of metalloproteinase-1 but higher MMP-1 and -7, which suggest a more favorable environment for tumor invasion and metastasis. Grade I and grade II hepatocellular carcinomas demonstrated high E-cadherin and decreased MMP-3 and 9, which may explain the rarity of extrahepatic metastasis in low-grade hepatocellular carcinomas despite the high circulatory volume of the liver. The histological progression from dysplastic nodule to well-differentiated hepatocellular carcinoma and to less differentiated tumors was associated with a gradual decrease in tissue expression of E-cadherin, tissue inhibitors of metalloproteinase-2 and -3. Metastatic hepatocellular carcinomas showed significantly lower level of tissue inhibitors of metalloproteinase-1, -2, -3 but higher level of MMP-7. These data suggest that tissue expression of E-cadherin, certain MMPs, and tissue inhibitors of metalloproteinases could be useful markers to predict the progression and metastasis of hepatocellular carcinoma. PMID- 16474380 TI - Expression of the CDK inhibitor p27kip1 and oxidative DNA damage in non neoplastic and neoplastic vulvar epithelial lesions. AB - Vulvar cancer represents an important medical problem worldwide whose incidence is increasing at an alarming rate in young females. Several factors have been linked to vulvar cancer development, but its exact pathogenesis remains to be determined. Vulvar tumorigenesis proceeds through intermediate dysplastic lesions, known as vulvar intraepithelial neoplasias, frequently associated with non-neoplastic epithelial disorders of the vulva, such as lichen sclerosus and squamous cell hyperplasia. In this study, the expression of the CDK inhibitor p27Kip1 and the extent of endogenous oxidative DNA damage were evaluated in vulvar specimens, including normal tissues, lichen sclerosus, squamous cell hyperplasia, vulvar intraepithelial neoplasias and invasive squamous cell carcinomas. We found that p27Kip1 was constantly expressed in normal vulvar epithelium cells while a progressive significant reduction in the percentage of p27Kip1-positive cells was observed in vulvar intraepithelial neoplasias (77%) and in invasive carcinomas (64%). Mean percentage of positive cells in invasive carcinomas, but not in vulvar intraepithelial neoplasias, was also significantly lower than squamous cell hyperplasia lesions (78%) while lichen sclerosus displayed a percentage of positive cells (45%) significantly lower than both vulvar intraepithelial neoplasias and invasive carcinomas. 8 hydroxydeoxyguanosine (8-OHdG) is considered a sensitive biomarker for oxidative stress. We observed a progressive significant increase in the levels of 8-OHdG and in the percentage of positive cells from normal vulvar epithelium to vulvar intraepithelial neoplasias (25%) and to invasive carcinomas (64%). Squamous cell hyperplasia displayed an intermediate percentage of positive cells comparable to vulvar intraepithelial neoplasias 2 but significantly higher than vulvar intraepithelial neoplasias 1 and lower than invasive carcinomas. Lichen sclerosus staining was significantly lower than carcinomas but higher than vulvar intraepithelial neoplasias and squamous cell hyperplasia. These results demonstrate that expression of p27Kip1 is downregulated while oxidative DNA damage increases from early non-neoplastic epithelial alterations through vulvar intraepithelial neoplasias to invasive vulvar carcinomas. Thus, both parameters might play an important role in the development of this cancer and their study might contribute to our understanding of human vulvar carcinogenesis. PMID- 16474382 TI - Reverse replay of behavioural sequences in hippocampal place cells during the awake state. AB - The hippocampus has long been known to be involved in spatial navigational learning in rodents, and in memory for events in rodents, primates and humans. A unifying property of both navigation and event memory is a requirement for dealing with temporally sequenced information. Reactivation of temporally sequenced memories for previous behavioural experiences has been reported in sleep in rats. Here we report that sequential replay occurs in the rat hippocampus during awake periods immediately after spatial experience. This replay has a unique form, in which recent episodes of spatial experience are replayed in a temporally reversed order. This replay is suggestive of a role in the evaluation of event sequences in the manner of reinforcement learning models. We propose that such replay might constitute a general mechanism of learning and memory. PMID- 16474381 TI - An shRNA barcode screen provides insight into cancer cell vulnerability to MDM2 inhibitors. AB - The identification of the cellular targets of small molecules with anticancer activity is crucial to their further development as drug candidates. Here, we present the application of a large-scale RNA interference-based short hairpin RNA (shRNA) barcode screen to gain insight in the mechanism of action of nutlin-3 (1). Nutlin-3 is a small-molecule inhibitor of MDM2, which can activate the p53 pathway. Nutlin-3 shows strong antitumor effects in mice, with surprisingly few side effects on normal tissues. Aside from p53, we here identify 53BP1 as a critical mediator of nutlin-3-induced cytotoxicity. 53BP1 is part of a signaling network induced by DNA damage that is frequently activated in cancer but not in healthy tissues. Our results suggest that nutlin-3's tumor specificity may result from its ability to turn a cancer cell-specific property (activated DNA damage signaling) into a weakness that can be exploited therapeutically. PMID- 16474383 TI - An in vitro fluorescence screen to identify antivirals that disrupt hepatitis B virus capsid assembly. AB - Virus assembly has not been routinely targeted in the development of antiviral drugs, in part because of the lack of tractable methods for screening in vitro. We have developed an in vitro assay of hepatitis B virus (HBV) capsid assembly, based on fluorescence quenching of dye-labeled capsid protein, for testing potential inhibitors. This assay is adaptable to high-throughput screening and can identify small-molecule inhibitors of virus assembly that prevent, inappropriately accelerate and/or misdirect capsid formation to yield aberrant particles. An in vitro primary screen has the advantage of identifying promising lead compounds affecting assembly without the requirement that they be taken up by cells in culture and be nontoxic. Our approach may facilitate the identification of antivirals targeting viruses other than HBV, such as avian influenza and HIV. PMID- 16474384 TI - A microtubule-binding domain in dynactin increases dynein processivity by skating along microtubules. AB - Microtubule-associated proteins (MAPs) use particular microtubule-binding domains that allow them to interact with microtubules in a manner specific to their individual cellular functions. Here, we have identified a highly basic microtubule-binding domain in the p150 subunit of dynactin that is only present in the dynactin members of the CAP-Gly family of proteins. Using single-particle microtubule-binding assays, we found that the basic domain of dynactin moves progressively along microtubules in the absence of molecular motors - a process we term 'skating'. In contrast, the previously described CAP-Gly domain of dynactin remains firmly attached to a single point on microtubules. Further analyses showed that microtubule skating is a form of one-dimensional diffusion along the microtubule. To determine the cellular function of the skating phenomenon, dynein and the dynactin microtubule-binding domains were examined in single-molecule motility assays. We found that the basic domain increased dynein processivity fourfold whereas the CAP-Gly domain inhibited dynein motility. Our data show that the ability of the basic domain of dynactin to skate along microtubules is used by dynein to maintain longer interactions for each encounter with microtubules. PMID- 16474385 TI - The polarity protein PAR-3 and TIAM1 cooperate in dendritic spine morphogenesis. AB - PAR-3 (partitioning-defective gene 3) is essential for cell polarization in many contexts, including axon specification. However, polarity proteins have not been implicated in later steps of neuronal differentiation, such as dendritic spine morphogenesis. Here, we show that PAR-3 is necessary for normal spine development in primary hippocampal neurons. Depletion of PAR-3 causes the formation of multiple filopodia- and lamellipodia-like dendritic protrusions - a phenotype similar to neurons expressing activated Rac. PAR-3 regulates spine formation by binding the Rac guanine nucleotide-exchange factor (GEF) TIAM1, and spatially restricting it to dendritic spines. Thus, a balance of PAR-3 and TIAM1 is essential to modulate Rac-GTP levels and to allow spine morphogenesis. PMID- 16474386 TI - Metabolic installation of thiols into sialic acid modulates adhesion and stem cell biology. AB - Metabolic 'oligosaccharide engineering' methods based on N-acetyl-D-mannosamine (ManNAc) analogs allow the glycocalyx of living cells to be remodeled. Herein we report the analog Ac(5)ManNTGc (1) that enables thiols to be expressed in surface sialic acids. By locating this versatile functional group on the outer periphery of normally nonadhesive human Jurkat cells, we obtained spontaneous cell-cell clustering and attachment to complementary maleimide-derivatized substrates. When analyzed in human embryoid body-derived (hEBD) stem cells, Ac(5)ManNTGc induced beta-catenin expression and altered cell morphology, consistent with neuronal differentiation. Notably, these effects were modulated by the growth substrate of the cells, with a stronger response observed on a gold surface than on glass. Together, these results establish sugar analogs as small-molecule tools for tissue engineering by providing a method for attaching cells to scaffolds via their surface carbohydrates as well as offering a means to influence stem cell fates. PMID- 16474387 TI - A functional genomics approach to the mode of action of apratoxin A. AB - The cyanobacterial metabolite apratoxin A (1) demonstrates potent cytotoxicity against tumor cell lines by a hitherto unknown mechanism. We have used functional genomics to elucidate the molecular basis for this activity. Gene expression profiling and DNA content analysis showed that apratoxin A induces G1-phase cell cycle arrest and apoptosis. Cell-based functional assays with a genome-wide collection of expression cDNAs showed that ectopic induction of fibroblast growth factor receptor (FGFR) signaling attenuates the apoptotic activity of apratoxin A. This natural product inhibited phosphorylation and activation of STAT3, a downstream effector of FGFR signaling. It also caused defects in FGF-dependent processes during zebrafish development, with concomitant reductions in expression levels of the FGF target gene mkp3. We conclude that apratoxin A mediates its antiproliferative activity through the induction of G1 cell cycle arrest and an apoptotic cascade, which is at least partially initiated through antagonism of FGF signaling via STAT3. PMID- 16474388 TI - Selective vulnerability and pruning of phasic motoneuron axons in motoneuron disease alleviated by CNTF. AB - Neurodegenerative diseases can have long preclinical phases and insidious progression patterns, but the mechanisms of disease progression are poorly understood. Because quantitative accounts of neuronal circuitry affected by disease have been lacking, it has remained unclear whether disease progression reflects processes of stochastic loss or temporally defined selective vulnerabilities of distinct synapses or axons. Here we derive a quantitative topographic map of muscle innervation in the hindlimb. We show that in two mouse models of motoneuron disease (G93A SOD1 and G85R SOD1), axons of fast-fatiguable motoneurons are affected synchronously, long before symptoms appear. Fast-fatigue resistant motoneuron axons are affected at symptom-onset, whereas axons of slow motoneurons are resistant. Axonal vulnerability leads to synaptic vesicle stalling and accumulation of BC12a1-a, an anti-apoptotic protein. It is alleviated by ciliary neurotrophic factor and triggers proteasome-dependent pruning of peripheral axon branches. Thus, motoneuron disease involves predictable, selective vulnerability patterns by physiological subtypes of axons, episodes of abrupt pruning in the target region and compensation by resistant axons. PMID- 16474389 TI - Dendritic patterning by Dscam and synaptic partner matching in the Drosophila antennal lobe. AB - In the olfactory system of Drosophila melanogaster, axons of olfactory receptor neurons (ORNs) and dendrites of second-order projection neurons typically target 1 of approximately 50 glomeruli. Dscam, an immunoglobulin superfamily protein, acts in ORNs to regulate axon targeting. Here we show that Dscam acts in projection neurons and local interneurons to control the elaboration of dendritic fields. The removal of Dscam selectively from projection neurons or local interneurons led to clumped dendrites and marked reduction in their dendritic field size. Overexpression of Dscam in projection neurons caused dendrites to be more diffuse during development and shifted their relative position in adulthood. Notably, the positional shift of projection neuron dendrites caused a corresponding shift of its partner ORN axons, thus maintaining the connection specificity. This observation provides evidence for a pre- and postsynaptic matching mechanism independent of precise glomerular positioning. PMID- 16474390 TI - Respiratory rhythm generation during gasping depends on persistent sodium current. AB - In severe hypoxia, homeostatic mechanisms maintain function of the brainstem respiratory network. We hypothesized that hypoxia involves a transition from neuronal mechanisms of normal breathing (eupnea) to a rudimentary pattern of inspiratory movements (gasping). We provide evidence for hypoxia-driven transformation within the central respiratory oscillator, in which gasping relies on persistent sodium current, whereas eupnea does not depend on this cellular mechanism. PMID- 16474391 TI - Visual stimuli-induced LTD of GABAergic synapses mediated by presynaptic NMDA receptors. AB - Local GABA (gamma-aminobutyric acid) circuits contribute to sensory experience dependent refinement of neuronal connections in the developing nervous system, but whether GABAergic synapses themselves can be rapidly modified by sensory stimuli is largely unknown. Here we report that repetitive light stimuli or theta burst stimulation (TBS) of the optic nerve in the developing Xenopus retinotectal system induces long-term potentiation (LTP) of glutamatergic inputs but long-term depression (LTD) of GABAergic inputs to the same tectal neuron. The LTD is due to a reduction in presynaptic GABA release and requires activation of presynaptic NMDA (N-methyl-D-aspartate) receptors (NMDARs) and coincident high-level GABAergic activity. Thus, the presynaptic NMDAR may function as a coincidence detector for adjacent glutamatergic and GABAergic activities, leading to coordinated synaptic modification by sensory experience. PMID- 16474392 TI - Decreases in the precision of Purkinje cell pacemaking cause cerebellar dysfunction and ataxia. AB - Episodic ataxia type-2 (EA2) is caused by mutations in P/Q-type voltage-gated calcium channels that are expressed at high densities in cerebellar Purkinje cells. Because P/Q channels support neurotransmitter release at many synapses, it is believed that ataxia is caused by impaired synaptic transmission. Here we show that in ataxic P/Q channel mutant mice, the precision of Purkinje cell pacemaking is lost such that there is a significant degradation of the synaptic information encoded in their activity. The irregular pacemaking is caused by reduced activation of calcium-activated potassium (K(Ca)) channels and was reversed by pharmacologically increasing their activity with 1-ethyl-2-benzimidazolinone (EBIO). Moreover, chronic in vivo perfusion of EBIO into the cerebellum of ataxic mice significantly improved motor performance. Our data support the hypothesis that the precision of intrinsic pacemaking in Purkinje cells is essential for motor coordination and suggest that K(Ca) channels may constitute a potential therapeutic target in EA2. PMID- 16474393 TI - The tempotron: a neuron that learns spike timing-based decisions. AB - The timing of action potentials in sensory neurons contains substantial information about the eliciting stimuli. Although the computational advantages of spike timing-based neuronal codes have long been recognized, it is unclear whether, and if so how, neurons can learn to read out such representations. We propose a new, biologically plausible supervised synaptic learning rule that enables neurons to efficiently learn a broad range of decision rules, even when information is embedded in the spatiotemporal structure of spike patterns rather than in mean firing rates. The number of categorizations of random spatiotemporal patterns that a neuron can implement is several times larger than the number of its synapses. The underlying nonlinear temporal computation allows neurons to access information beyond single-neuron statistics and to discriminate between inputs on the basis of multineuronal spike statistics. Our work demonstrates the high capacity of neural systems to learn to decode information embedded in distributed patterns of spike synchrony. PMID- 16474394 TI - Structural basis for a major histocompatibility complex class Ib-restricted T cell response. AB - In contrast to antigen-specific immunity orchestrated by major histocompatibility complex (MHC) class Ia molecules, the ancestrally related nonclassical MHC class Ib molecules generally mediate innate immune responses. Here we have demonstrated the structural basis by which the MHC class Ib molecule HLA-E mediates an adaptive MHC-restricted cytotoxic T lymphocyte response to human cytomegalovirus. Highly constrained by host genetics, the response showed notable fine specificity for position 8 of the viral peptide, which is the sole discriminator of self versus nonself. Despite the evolutionary divergence of MHC class Ia and class Ib molecules, the structure of the T cell receptor-MHC class Ib complex was very similar to that of conventional T cell receptor-MHC class Ia complexes. These results emphasize the evolutionary 'ambiguity' of HLA-E, which not only interacts with innate immune receptors but also has the functional capacity to mediate virus-specific cytotoxic T lymphocyte responses during adaptive immunity. PMID- 16474395 TI - Positive regulation of immune cell function and inflammatory responses by phosphatase PAC-1. AB - Mitogen-activated protein kinases facilitate many cellular processes and are essential for immune cell function. Their activity is controlled by kinases and dual-specificity phosphatases. A comprehensive microarray analysis of human leukocytes identified DUSP2 (encoding the phosphatase PAC-1) as one of the most highly induced transcripts in activated immune cells. We generated Dusp2(-/-) mice and found considerably reduced inflammatory responses in the 'K/BxN' model of rheumatoid arthritis. PAC-1 deficiency led to increased activity of Jun kinase (Jnk) but unexpected impairment of the activity of extracellular signal-regulated kinase (Erk) and the kinase p38, reduced activity of the transcription factor Elk1 and a complex of mobilized transcription factor NFAT and the AP-1 transcription factor and decreased effector immune cell function. Thus, PAC-1 is a key positive regulator of inflammatory cell signaling and effector functions, mediated through Jnk and Erk mitogen-activated protein kinase crosstalk. PMID- 16474398 TI - A novel peptide CXCR ligand derived from extracellular matrix degradation during airway inflammation. AB - We describe the tripeptide neutrophil chemoattractant N-acetyl Pro-Gly-Pro (PGP), derived from the breakdown of extracellular matrix (ECM), which shares sequence and structural homology with an important domain on alpha chemokines. PGP caused chemotaxis and production of superoxide through CXC receptors, and administration of peptide caused recruitment of neutrophils (PMNs) into lungs of control, but not CXCR2-deficient mice. PGP was generated in mouse lung after exposure to lipopolysaccharide, and in vivo and in vitro blockade of PGP with monoclonal antibody suppressed PMN responses as much as chemokine-specific monoclonal antibody. Extended PGP treatment caused alveolar enlargement and right ventricular hypertrophy in mice. PGP was detectable in substantial concentrations in a majority of bronchoalveolar lavage samples from individuals with chronic obstructive pulmonary disease, but not control individuals. Thus, PGP's activity links degradation of ECM with neutrophil recruitment in airway inflammation, and PGP may be a biomarker and therapeutic target for neutrophilic inflammatory diseases. PMID- 16474399 TI - Interleukin-15 rescues tolerant CD8+ T cells for use in adoptive immunotherapy of established tumors. AB - CD8+ T cells can mediate eradication of established tumors, and strategies to amplify tumor-reactive T-cell numbers by immunization or ex vivo expansion followed by adoptive transfer are currently being explored in individuals with cancer. Generating effective CD8+ T cell-mediated responses to tumors is often impeded by T-cell tolerance to relevant tumor antigens, as most of these antigens are also expressed in normal tissues. We examined whether such tolerant T cells could be rescued and functionally restored for use in therapy of established tumors. We used a transgenic T-cell receptor (TCR) mouse model in which peripheral CD8+ T cells specific for a candidate tumor antigen also expressed in liver are tolerant, failing to proliferate or secrete interleukin (IL)-2 in response to antigen. Molecular and cellular analysis showed that these tolerant T cells expressed the IL-15 receptor alpha chain, and could be induced to proliferate in vitro in response to exogenous IL-15. Such proliferation abrogated tolerance and the rescued cells became effective in treating leukemia. Therefore, high-affinity CD8+ T cells are not necessarily deleted by encounter with self antigen in the periphery, and can potentially be rescued and expanded for use in tumor immunotherapy. PMID- 16474400 TI - Successful transduction of liver in hemophilia by AAV-Factor IX and limitations imposed by the host immune response. AB - We have previously shown that a single portal vein infusion of a recombinant adeno-associated viral vector (rAAV) expressing canine Factor IX (F.IX) resulted in long-term expression of therapeutic levels of F.IX in dogs with severe hemophilia B. We carried out a phase 1/2 dose-escalation clinical study to extend this approach to humans with severe hemophilia B. rAAV-2 vector expressing human F.IX was infused through the hepatic artery into seven subjects. The data show that: (i) vector infusion at doses up to 2 x 10(12) vg/kg was not associated with acute or long-lasting toxicity; (ii) therapeutic levels of F.IX were achieved at the highest dose tested; (iii) duration of expression at therapeutic levels was limited to a period of approximately 8 weeks; (iv) a gradual decline in F.IX was accompanied by a transient asymptomatic elevation of liver transaminases that resolved without treatment. Further studies suggested that destruction of transduced hepatocytes by cell-mediated immunity targeting antigens of the AAV capsid caused both the decline in F.IX and the transient transaminitis. We conclude that rAAV-2 vectors can transduce human hepatocytes in vivo to result in therapeutically relevant levels of F.IX, but that future studies in humans may require immunomodulation to achieve long-term expression. PMID- 16474401 TI - Disruption of PTEN coupling with 5-HT2C receptors suppresses behavioral responses induced by drugs of abuse. AB - The widespread distribution of the tumor suppressor PTEN (phosphatase and tensin homolog deleted on chromosome 10) in the adult brain suggests its role in a broad range of brain functions. Here we show evidence supporting a physical interaction of PTEN with a region in the third intracellular loop (3L4F) of the serotonin 5 HT2C receptor (5-HT2cR, formerly 5-HT1c receptor) in cell cultures. PTEN limits agonist-induced phosphorylation of 5-HT2cR through its protein phosphatase activity. We showed the probable existence of PTEN:5-HT2cR complexes in putative dopaminergic neurons in the rat ventral tegmental area (VTA), a brain region in which virtually all abused drugs exert rewarding effects by activating its dopamine neurons. We synthesized the interfering peptide Tat-3L4F, which is able to disrupt PTEN coupling with 5-HT2cR. Systemic application of Tat-3L4F or the 5 HT2cR agonist Ro600175 suppressed the increased firing rate of VTA dopaminergic neurons induced by delta9-tetrahydrocannabinol (THC), the psychoactive ingredient of marijuana. Using behavioral tests, we found that Tat-3L4F or Ro600175 blocks conditioned place preference of THC or nicotine, and that Ro600175, but not Tat 3L4F, produces anxiogenic effects, penile erection, hypophagia and motor functional suppression. These results suggest a potential strategy for treating drug addiction with the Tat-3L4F peptide. PMID- 16474402 TI - Molecular recognition of p53 and MDM2 by USP7/HAUSP. AB - The ubiquitin-specific protease, USP7, has key roles in the p53 pathway whereby it stabilizes both p53 and MDM2. We show that the N-terminal domain of USP7 binds two closely spaced 4-residue sites in both p53 and MDM2, falling between p53 residues 359-367 and MDM2 residues 147-159. Cocrystal structures with USP7 were determined for both p53 peptides and for one MDM2 peptide. These peptides bind the same surface of USP7 as Epstein-Barr nuclear antigen-1, explaining the competitive nature of the interactions. The structures and mutagenesis data indicate a preference for a P/AXXS motif in peptides that bind USP7. Contacts made by serine are identical and crucial for all peptides, and Trp165 in the peptide-binding pocket of USP7 is also crucial. These results help to elucidate the mechanism of substrate recognition by USP7 and the regulation of the p53 pathway. PMID- 16474403 TI - Intermediates revealed in the kinetic mechanism for DNA unwinding by a monomeric helicase. AB - Helicases unwind dsDNA during replication, repair and recombination in an ATP dependent reaction. The mechanism for helicase activity can be studied using oligonucleotide substrates to measure formation of single-stranded (ss) DNA from double-stranded (ds) DNA. This assay provides an 'all-or-nothing' readout because partially unwound intermediates are not detected. We have determined conditions under which an intermediate in the reaction cycle of Dda helicase can be detected by trapping a partially unwound substrate. The appearance of this intermediate supports a model in which each ssDNA product interacts with the helicase after unwinding has occurred. Kinetic analysis indicates that the intermediate appears during a slow step in the reaction cycle that is flanked by faster steps for unwinding. These observations demonstrate a complex mechanism containing nonuniform steps for a monomeric helicase. The potential biological significance of such a mechanism is discussed. PMID- 16474404 TI - Germline KRAS and BRAF mutations in cardio-facio-cutaneous syndrome. AB - Cardio-facio-cutaneous (CFC) syndrome is characterized by a distinctive facial appearance, heart defects and mental retardation. It phenotypically overlaps with Noonan and Costello syndrome, which are caused by mutations in PTPN11 and HRAS, respectively. In 43 individuals with CFC, we identified two heterozygous KRAS mutations in three individuals and eight BRAF mutations in 16 individuals, suggesting that dysregulation of the RAS-RAF-ERK pathway is a common molecular basis for the three related disorders. PMID- 16474405 TI - Germline KRAS mutations cause Noonan syndrome. AB - Noonan syndrome (MIM 163950) is characterized by short stature, facial dysmorphism and cardiac defects. Heterozygous mutations in PTPN11, which encodes SHP-2, cause approximately 50% of cases of Noonan syndrome. The SHP-2 phosphatase relays signals from activated receptor complexes to downstream effectors, including Ras. We discovered de novo germline KRAS mutations that introduce V14I, T58I or D153V amino acid substitutions in five individuals with Noonan syndrome and a P34R alteration in a individual with cardio-facio-cutaneous syndrome (MIM 115150), which has overlapping features with Noonan syndrome. Recombinant V14I and T58I K-Ras proteins show defective intrinsic GTP hydrolysis and impaired responsiveness to GTPase activating proteins, render primary hematopoietic progenitors hypersensitive to growth factors and deregulate signal transduction in a cell lineage-specific manner. These studies establish germline KRAS mutations as a cause of human disease and infer that the constellation of developmental abnormalities seen in Noonan syndrome spectrum is, in large part, due to hyperactive Ras. PMID- 16474407 TI - Rhythmic CLOCK-BMAL1 binding to multiple E-box motifs drives circadian Dbp transcription and chromatin transitions. AB - Mammalian circadian rhythms are based on transcriptional and post-translational feedback loops. Essentially, the activity of the transcription factors BMAL1 (also known as MOP3) and CLOCK is rhythmically counterbalanced by Period (PER) and Cryptochrome (CRY) proteins to govern time of day-dependent gene expression. Here we show that circadian regulation of the mouse albumin D element-binding protein (Dbp) gene involves rhythmic binding of BMAL1 and CLOCK and marked daily chromatin transitions. Thus, the Dbp transcription cycle is paralleled by binding of BMAL1 and CLOCK to multiple extra- and intragenic E boxes, acetylation of Lys9 of histone H3, trimethylation of Lys4 of histone H3 and a reduction of histone density. In contrast, the antiphasic daily repression cycle is accompanied by dimethylation of Lys9 of histone H3, the binding of heterochromatin protein 1alpha and an increase in histone density. The rhythmic conversion of transcriptionally permissive chromatin to facultative heterochromatin relies on the presence of functional BMAL1-CLOCK binding sites. PMID- 16474406 TI - Feedback repression is required for mammalian circadian clock function. AB - Direct evidence for the requirement of transcriptional feedback repression in circadian clock function has been elusive. Here, we developed a molecular genetic screen in mammalian cells to identify mutants of the circadian transcriptional activators CLOCK and BMAL1, which were uncoupled from CRYPTOCHROME (CRY)-mediated transcriptional repression. Notably, mutations in the PER-ARNT-SIM domain of CLOCK and the C terminus of BMAL1 resulted in synergistic insensitivity through reduced physical interactions with CRY. Coexpression of these mutant proteins in cultured fibroblasts caused arrhythmic phenotypes in population and single-cell assays. These data demonstrate that CRY-mediated repression of the CLOCK/BMAL1 complex activity is required for maintenance of circadian rhythmicity and provide formal proof that transcriptional feedback is required for mammalian clock function. PMID- 16474408 TI - Facial rash mimicking cutaneous acute graft-versus-host disease after allogeneic stem cell transplantation for osteomyelofibrosis - was Demodex the culprit? PMID- 16474409 TI - Differential effect of conditioning regimens on cytokine responses during allogeneic stem cell transplantation. AB - The purpose of this study was to characterize cytokine responses during conditioning in patients undergoing allogeneic stem cell transplantation (SCT) with the aim to identify which markers that may reliably reflect inflammatory activity during conditioning. We investigated inflammatory and anti-inflammatory mediators in plasma samples drawn daily during the conditioning of 20 patients. Soluble tumour necrosis factor alpha receptor I (sTNFRI) increased during the conditioning reflecting the type of conditioning given. Antithymocyte globulin (ATG) was the most potent inducer of sTNFRI (288% increase (median) P=0.002), followed by VP-16 (184%, P=0.03), cyclophosphamide (129%, P=0.03) and total body irradiation (148%, P=0.0005). Administration of i.v. busulfan (Busilvex; BU) was not associated with significant changes in sTNFRI levels. At day 0 (the day of stem cell infusion) the sTNFRI levels were not only elevated compared with baseline (188% increase), P<0.0001), they also correlated with the baseline values (r=0.72, P=0.0003). The levels of tumour necrosis factor, interleukin (IL) 1beta, IL-6, IL-8, IL-10 and IL-12 stayed at low levels during the conditioning, except from a transient increase in the levels of IL-6, IL-8, IL-10 and IL- receptor antagonist (IL-Ra) seen after ATG infusion. These findings suggest that further investigation of circulating sTNFRI levels may be of interest in studies of prognostic factors in SCT. PMID- 16474410 TI - Allogeneic stem cell transplantation after reduced-intensity conditioning in a patient with T-cell prolymphocytic leukemia: graft-versus-tumor effect and long term remission. PMID- 16474411 TI - Pharmacological insights obtained from structure-function studies of ionotropic glutamate receptors. AB - Ionotropic glutamate receptors mediate the vast majority of fast excitatory synaptic transmission in the CNS. Elucidating the structure of these proteins is central to understanding their overall function and in the last few years a tremendous amount of knowledge has been gained from the crystal structures of the ligand-binding domains of the receptor protein. These efforts have enabled us to unravel the possible mechanisms of partial agonism, agonist selectivity and desensitization. This review summarizes recent data obtained from structural studies of the binding pockets of the GluR2, GluR5/6, NR1 and NR2A subunits and discusses these studies together with homology modelling and molecular dynamics simulations that have suggested possible binding modes for full and partial agonists as well as antagonists within the binding pocket of various ionotropic glutamate receptor subunits. Comparison of the ligand-binding pockets suggests that the ligand-binding mechanisms may be conserved throughout the glutamate receptor family, although agonist selectivity may be explained by a number of features inherent to the AMPA, kainate and NMDA receptor-binding pockets such as steric occlusion, cavity size and the contribution of water-bridged interactions. PMID- 16474412 TI - Cannabinoid agonists induce relaxation in the bovine ophthalmic artery: evidences for CB1 receptors, nitric oxide and potassium channels. AB - Glaucoma pathophysiology appears to involve vascular deficits, which may contribute to initiation and progression of the disease. Anandamide, the endogenous cannabinoid ligand, and WIN55212-2, a synthetic cannabinoid agonist, are able to evoke concentration-dependent relaxations in bovine ophthalmic artery rings, precontracted with 5-hydroxytryptamine (5-HT) (1 microM). Endothelium removal reduces cannabinoid agonist potency and efficacy. The selective cannabinoid 1 (CB1) receptor antagonists SR141716A (100 nM) and AM251 (100 nM) cause a shift to the right in the concentration-response curves to anandamide and WIN55212-2 in arterial rings both in the presence and in the absence of endothelium. In endothelium-intact arteries, the nitric oxide synthase inhibitor, N(G)-monomethyl-L-arginine (L-NMMA, 300 microM), completely blocked the anandamide- and WIN55212-2-relaxant responses; by contrast, the nitric oxide donor S-nitroso-N-acetylpenicillamine (SNAP, 100 microM) induced an increase in vasorelaxant responses to cannabinoid agonists. Relaxations to anandamide and WIN55212-2 were inhibited by iberiotoxin (IbTX, 200 nM), a blocker of large conductance, Ca2+-activated K+ channel (BK(Ca)), and by 4-aminopyridine (4-AP; 1 mM), a blocker of delayed rectifier K+ channel, whereas the blockade of K(ATP) channels by glibenclamide (5 microM) and of small conductance Ca2+-activated K+ channels (SK(Ca)) by apamin (100 nM) did not produce any effects. These data suggest that anandamide and WIN55212-2 relax the bovine ophthalmic artery by involving CB1 the cannabinoid receptor-sensitive pathway. In endothelium-intact arteries, relaxation occurs through activation of nitric oxide synthase cyclic GMP and Ca2+-activated K+ channels. They also cause endothelium-independent relaxation by involving potassium channel opening. PMID- 16474413 TI - The proconvulsant effect of sildenafil in mice: role of nitric oxide-cGMP pathway. AB - Recent evidence indicates that sildenafil may exert some central effects through enhancement of nitric oxide (NO)-mediated effects. NO is known to have modulatory effects on seizure threshold, raising the possibility that sildenafil may alter seizure susceptibility through NO-mediated mechanisms. This study was performed to examine whether sildenafil influences the threshold of clonic and/or generalized tonic seizures through modulation of nitric oxide (NO)-cGMP pathway. The effect of sildenafil (1-40 mg kg(-1)) was investigated on clonic seizures induced by intravenous administration of GABA antagonists pentylenetetrazole (PTZ) and bicuculine and on generalized tonic seizures induced by intraperitoneal administration of high dose PTZ in male Swiss mice. The interaction of sildenafil induced effects with NO-cGMP pathway was examined using nitric oxide synthase (NOS) inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME), NOS substrate L arginine, NO donor, sodium nitroprusside (SNP) and guanylyl cyclase inhibitor methylene blue (MB). Sildenafil induced a dose-dependent proconvulsant effect in both models of clonic, but not generalized tonic type of seizures. Pretreatment with either MB or L-NAME inhibited the proconvulsant effect of sildenafil, indicating the mediation of this effect by NO-cGMP pathway. In addition, a subeffective dose of sildenafil induced an additive proconvulsant effect when combined with either L-arginine or SNP. Sildenafil induces a proconvulsant effect on clonic seizure threshold that interacts with both exogenously and endogenously released NO and may be linked to activation of NO-cGMP pathway. PMID- 16474414 TI - ROS-sensitive cytochrome P450 activity maintains endothelial dilatation in ageing but is transitory in dyslipidaemic mice. AB - Risk factors for cardiovascular diseases (CVD) have been proposed to accelerate the vascular endothelial dysfunction that develops during the normal ageing process. The objective of this work was to study the impact of dyslipidaemia (DL) on the dilatory efficacy of the non-NO/non-PGI2 endothelium-derived hyperpolarising factor (EDHF) through maturation and ageing. We isolated and pressurised (80 mmHg) gracilis arterial segments from 3, 12 and 20-month-old (m/o) DL mice expressing the human apolipoprotein B-100 and wild-type (WT) C57BL/6 mice. EDHF-dependent dilatations to acetylcholine (ACh) were measured in the presence of L-NNA (100 microM, NOS inhibitor) and indomethacin (INDO; 10 microM, COX inhibitor). Data are expressed as mean+/-s.e.m.EDHF-mediated maximal dilatation of arteries isolated from WT mice declined by 44% with ageing, from 86+/-3% at 3 months to 66+/-8% at 12 and 48+/-4% at 20 months of age (P<0.05). This decline was magnified by DL to 73%, characterised by an early increased efficacy at 3 m/o (95+/-2%, P<0.05) and a worsening of the dysfunction at 20 m/o (26+/-2%, P<0.05). 17-Octadecynoic acid (17-ODYA), a cytochrome P450/epoxygenase inhibitor, reduced by 56% (P<0.05) ACh-induced EDHF-dependent dilatation of arteries isolated from 3 m/o DL--but not WT--mice, an effect of 17-ODYA disappearing in older DL mice. 17-ODYA, however, reduced (P<0.05) ACh-induced EDHF-dependent dilatation in arteries isolated from 12 m/o WT mice by 35% and from 20 m/o WT mice by 31% (P<0.05). Reactive oxygen species production was increased in arteries isolated from 12 m/o DL mice. The antioxidant N-acetyl-L cystein (NAC) restored the 17-ODYA-sensitive responses in arteries isolated from 12 - but not 20 - m/o DL mice (84+/-3% from an E(max) of 57+/-8%; P<0.05). NAC did not affect the dilatation of arteries isolated from WT mice. Our data suggest that the decline in EDHF-dependent dilatation is hastened by DL despite the early expression of a 17-ODYA-sensitive pathway increasing the efficacy of the non NO/non-PGI2 endothelium-dependent dilatation. Acute free radical production contributes to the endothelial dysfunction in the presence of DL only, by abrogating this latter pathway. This 17-ODYA-sensitive pathway, however, appears in 12 m/o WT mice and remains active at 20 m/o. PMID- 16474415 TI - Mechanism of hERG K+ channel blockade by the fluoroquinolone antibiotic moxifloxacin. AB - The fluoroquinolone antibiotic moxifloxacin has been associated with the acquired long QT syndrome and is used as a positive control in the evaluation of the QT interval prolonging potential of new drugs. In common with other QT-prolonging agents, moxifloxacin is known to inhibit the hERG potassium K+ channel, but at present there is little mechanistic information available on this action. This study was conducted in order to characterise the inhibition of hERG current (I(hERG)) by moxifloxacin, and to determine the role in drug binding of the S6 aromatic amino-acid residues Tyr652 and Phe656. hERG currents were studied using whole-cell patch clamp (at room temperature and at 35-37 degrees C) in an HEK293 cell line stably expressing hERG channels. Moxifloxacin reversibly inhibited currents in a dose-dependent manner. We investigated the effects of different voltage commands to elicit hERG currents on moxifloxacin potency. Using a 'step ramp' protocol, the IC50 was 65 microM at room temperature and 29 microM at 35 degrees C. When a ventricular action potential waveform was used to elicit currents, the IC50 was 114 microM. Block of hERG by moxifloxacin was found to be voltage-dependent, occurred rapidly and was independent of stimulation frequency. Mutagenesis of the S6 helix residue Phe656 to Ala failed to eliminate or reduce the moxifloxacin-mediated block whereas mutation of Tyr652 to Ala reduced moxifloxacin block by approximately 66%. Our data demonstrate that moxifloxacin blocks the hERG channel with a preference for the activated channel state. The Tyr652 but not Phe656 S6 residue is involved in moxifloxacin block of hERG, concordant with an interaction in the channel inner cavity. PMID- 16474416 TI - Species-dependent smooth muscle contraction to Neuromedin U and determination of the receptor subtypes mediating contraction using NMU1 receptor knockout mice. AB - The peptide ligand neuromedin U (NMU) has been implicated in an array of biological activities, including contraction of uterine, intestinal and urinary bladder smooth muscle. However, many of these responses appear to be species specific. This study was undertaken to fully elucidate the range of smooth muscle stimulating effects of NMU in rats, mice and guinea-pigs, and to examine the extent of the species differences. In addition, the NMU1 receptor knockout mouse was used to determine which receptor subtype mediates the contractile responses generated by NMU in the mouse. A range of isolated organ in vitro bioassays were carried out, which were chosen to re-confirm previous literature reports (uterine and stomach fundus contraction) and also to explore potentially novel smooth muscle responses to NMU. This investigation uncovered a number of previously unidentified NMU-mediated responses: contraction of rat lower esophageal sphinster (LES), rat ileum, mouse gallbladder, enhancement of electrically evoked contractions in rat and mouse vas deferens, and a considerable degree of cross species differences. Studies using the NMU1 receptor knockout mice revealed that in the mouse fundus and gallbladder assays the NMU contractile response was mediated entirely through the NMU1 receptor subtype, whereas, in assays of mouse uterus and vas deferens, the response to NMU was unchanged in the NMU1 receptor knockout mouse, suggesting that the NMU response may be mediated through the NMU2 receptor subtype. NMU receptor subtype-selective antagonists are required to further elucidate the role of the individual receptor subtypes. PMID- 16474417 TI - Downregulation of propranolol-sensitive beta-adrenoceptor signaling after inhibition of nitric oxide synthesis. AB - The beta-adrenoceptor agonist, isoprenaline, elicits vasodilation and tachycardia in anesthetized rats via activation of propranolol-sensitive beta1- and beta2 adrenoceptors and also by propranolol-insensitive beta1- and beta3-adrenoceptors. The aim of this study was to determine whether the relative contribution of propranolol-sensitive and -insensitive beta-adrenoceptors to the changes in heart rate (HR) and vascular resistances elicited by isoprenaline is altered after blockade of nitric oxide (NO) synthase, in pentobarbital-anesthetized rats. The hemodynamic responses elicited by isoprenaline (0.1 and 0.5 microg kg(-1), i.v.) were determined before and after injection of saline or the NO synthase inhibitor, N(G)-nitro-L-arginine methylester (L-NAME, 50 micromol kg(-1), i.v.), and again after injection of the beta1- and beta2-adrenoceptor antagonist, propranolol (1 mg kg(-1), i.v.). The responses elicited by the above doses of isoprenaline were also determined before and during infusion of the alpha1 adrenoceptor agonist, phenylephrine (3 microg kg(-1) min(-1), i.v.), and again 15 20 min after injection of propranolol (1.0 mg kg(-1), i.v.). Both doses of isoprenaline elicited tachycardia and reductions in vascular resistances. Propranolol eliminated the responses elicited by the lower dose of isoprenaline and substantially diminished the responses elicited by the higher dose of the beta1-, beta2- and beta3-adrenoceptor agonist. The maximal vasodilator responses elicited by both doses of isoprenaline were not diminished whereas the maximal increases in HR were higher after injection of L-NAME. The ability of propranolol to diminish the hemodynamic actions of isoprenaline was substantially diminished in L-NAME-treated rats, whereas propranolol retained its potency in rats that received an equi-pressor infusion of the alpha1-adrenoceptor agonist, phenylephrine. The finding that the maximal vasodilator responses elicited by isoprenaline were not diminished by L-NAME suggests that the vasodilation elicited by this drug was due to direct activation of beta-adrenoceptors on vascular smooth muscle and that the full compliment of isoprenaline-sensitive receptors was not changed after inhibition of NO synthesis. However, these results suggest that the activities of propranolol-sensitive beta-adrenoceptors are downregulated, whereas propranolol-insensitive beta-adrenoceptors are upregulated upon the loss of exposure to endothelial nitrosyl factors. PMID- 16474418 TI - Agonist-specific activation of the beta2-adrenoceptor/Gs-protein and beta2 adrenoceptor/Gi-protein pathway in adult rat ventricular cardiomyocytes. AB - In rat ventricular cardiomyocytes beta2-adrenoceptors (AR) couple to Gs- and Gi protein, and evidence has accumulated that beta2-AR agonists can differentially activate either Gs- or Gs- and Gi-protein. In this study, in isolated adult rat ventricular cardiomyocytes, we assessed the effects of pertussis toxin (PTX) on beta2-AR agonist (terbutaline (TER), salbutamol (SAL) and fenoterol (FEN)) evoked inhibition of phenylephrine (PE)-induced increase in the rate of protein synthesis (assessed as [3H]phenylalanine incorporation) to find out which beta2 AR agonist activates selectively Gs- or Gs- and Gi-protein. PE (1 microM) increased the rate of protein synthesis from 100% to 130+/-2% (n = 34). FEN, TER and SAL (1 nM-10 microM) inhibited PE-induced increase in the rate of protein synthesis concentration-dependently. FEN inhibited PE effects almost completely (from 132+/-3 to 101+/-1%), whereas TER and SAL caused only partial inhibition (from 131+/-2 to 114+/-2 and 129+/-1 to 111+/-2%, respectively). Pretreatment of cardiomyocytes with PTX (250 ng ml(-1) for 16 h at 37 degrees C) did not affect FEN effects, but converted TER- and SAL-evoked partial inhibition into complete inhibition. Inhibitory effects of the three beta2-AR agonists were markedly attenuated by beta1-AR selective antagonist CGP 20712A (CGP) (300 nM); in contrast, beta2-AR selective antagonist ICI 118,551 (55 nM) inhibited the inhibitory effects of the three beta2-AR agonists only in PTX-pretreated cardiomyocytes,with beta1-AR blocked by CGP. We conclude that, in adult rat ventricular cardiomyocytes, FEN activates selectively the Gs protein-pathway, while TER and SAL activate the Gs- and Gi-protein pathways. Part of the effects of these three beta2-AR agonists appears to be mediated by beta1-AR. PMID- 16474419 TI - A novel steroid-like compound F90927 exerting positive-inotropic effects in cardiac muscle. AB - Here we report a novel steroid-like compound F90363, exhibiting positive inotropy in vivo and in vitro in various cardiac muscle preparations. F90363 is a racemic mixture composed of the stereoisomers (-)-F90926 and (+)-F90927. Only F90927 exerted positive inotropy, while F90926 induced a weak negative inotropy, but only at concentrations 10(3) times higher than F90927 and most likely resulting from an unspecific interaction. The rapid time course of the action of F90927 suggested a direct interaction with a cellular target rather than a genomic alteration. We could identify the L-type Ca2+ current I(Ca(L)) as a main target of F90927, while excluding other components of cardiac Ca2+ signalling as potential contributors. In addition, several other signaling pathways known to lead to positive inotropy (e.g. alpha- and beta-adrenergic stimulation, cAMP pathways) could be excluded as targets of F90927. However, vessel contraction and stiffening of the cardiac muscle at high doses (>30 microM, 0.36 mg kg(-1), respectively) prevent the use of F90927 as a candidate for drug development. Since the compound may still find valuable applications in research, the aim of the present study was to identify the cellular target and the mechanism of inotropy of F90927. PMID- 16474420 TI - Mixed beta3-adrenoceptor agonist and alpha1-adrenoceptor antagonist properties of nebivolol in rat thoracic aorta. AB - Nebivolol, a selective beta-adrenoceptor (beta1-AR) antagonist, induces vasodilatation by an endothelium- and NO-cGMP-dependent pathway. However, the mechanisms involved in the vascular effect of nebivolol have not been established. Thus, we evaluated the role of alpha1 and beta3-ARs in nebivolol induced vasodilatation. The responses to nebivolol were investigated in vitro in thoracic aortic rings isolated from male Sprague-Dawley rats. Nebivolol (0.1-10 microM) significantly shifted the concentration-response curve to phenylephrine, an alpha1-AR agonist, to the right in a concentration-dependent manner (pA2 = 6.5). Conversely, the concentration-response curve to endothelin 1 (ET1) was unaffected by nebivolol. In ET1-precontracted rings, nebivolol induced a concentration-dependent relaxation, which was unaffected by nadolol (a beta1/beta2-AR antagonist) but was significantly reduced by L-748,337 (a beta3-AR antagonist), endothelium removal or pretreatment with L-NMMA (an NOS inhibitor). Similar results were obtained with a beta3-AR agonist, SR 58611A. It was concluded that, in rat aorta, nebivolol-induced relaxation results from both inhibition of alpha1-ARs and activation of beta3-ARs. In addition, we confirmed that the endothelium and the NO pathway are involved in the vascular effect of nebivolol. The identification of these vascular targets of nebivolol indicate that it has therapeutic potential for the treatment of pathological conditions associated with an elevation of sympathetic tone, such as heart failure and hypertension. PMID- 16474421 TI - Aminoglutethimide prevents excitotoxic and ischemic injuries in cortical neurons. AB - Aminoglutethimide is a clinically available drug that suppresses steroid biosynthesis by inhibiting enzymes such as cytochrome P450scc and aromatase. Because several members of neurosteroids regulate glutamate receptors, we investigated the effect of aminoglutethimide on cell death induced by overactivation of glutamate receptors in CNS neurons. Long-term pretreatment of organotypic cerebrocortical slice cultures with aminoglutethimide (100-1000 microM) for 6 days or over resulted in concentration-dependent suppression of neuronal cell death induced by NMDA. Aminoglutethimide (1000 microM) also inhibited neurotoxicity of AMPA and kainate, but not of ionomycin or staurosporine. The protective effect of aminoglutethimide against NMDA cytotoxicity was not mimicked by other steroid synthesis inhibitors including trilostane and exemestane, and was not reversed by concurrent application of steroids such as pregnenolone, estrone, 17beta-estradiol and estriol. In dissociated rat cerebrocortical cell cultures, long-term treatment with aminoglutethimide (10-1000 microM) attenuated NMDA receptor-mediated glutamate cytotoxicity but produced no significant effect on glutamate-induced increases in intracellular Ca2+. Brief as well as long-term pretreatment with aminoglutethimide (30-1000 microM) prevented NMDA receptor-dependent ischemic neuronal injury in organotypic cerebrocortical slice cultures, which was associated with suppression of glutamate release during the ischemic insult. These results indicate that aminoglutethimide, irrelevant to its actions on neurosteroid synthesis, protects CNS neurons from excitotoxic and ischemic injuries. Development of aminoglutethimide analogs possessing neuroprotective properties may be of therapeutic value. PMID- 16474424 TI - Granulocyte-macrophage colony-stimulating factor (GM-CSF) and T-cell responses: what we do and don't know. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) is an important hematopoietic growth factor and immune modulator. GM-CSF also has profound effects on the functional activities of various circulating leukocytes. It is produced by a variety of cell types including T cells, macrophages, endothelial cells and fibroblasts upon receiving immune stimuli. Although GM-CSF is produced locally, it can act in a paracrine fashion to recruit circulating neutrophils, monocytes and lymphocytes to enhance their functions in host defense. Recent intensive investigations are centered on the application of GM-CSF as an immune adjuvant for its ability to increase dendritic cell (DC) maturation and function as well as macrophage activity. It is used clinically to treat neutropenia in cancer patients undergoing chemotherapy, in AIDS patients during therapy, and in patients after bone marrow transplantation. Interestingly, the hematopoietic system of GM-CSF-deficient mice appears to be normal; the most significant changes are in some specific T cell responses. Although molecular cloning of GM CSF was carried out using cDNA library of T cells and it is well known that the T cells produce GM-CSF after activation, there is a lack of systematic investigation of this cytokine in production by T cells and its effect on T cell function. In this article, we will focus mainly on the immunobiology of GM-CSF in T cells. PMID- 16474422 TI - The effects of resveratrol, a phytoalexin derived from red wines, on chronic inflammation induced in an experimentally induced colitis model. AB - Neutrophil infiltration, proinflammatory cytokines, eicosanoid generation and oxidative stress have been implicated in colitis. Resveratrol is a polyphenolic compound found in grapes and wine, with multiple pharmacological actions, including anti-inflammatory, antioxidant, antitumour and immunomodulatory activities. In a previous report, we documented that resveratrol decreases the degree of inflammation associated with acute experimental colonic inflammation, but its effects on chronic experimental colitis remain undetermined. The aim of this research was to investigate the effects of resveratrol on the chronic colonic injury caused by intracolonic instillation of trinitrobenzenesulphonic acid (TNBS) in rats. The inflammatory response was assessed by histology and myeloperoxidase activity. Tumour necrosis factor alpha (TNF-alpha) production, histological and histochemical analysis of the lesions were also carried out. We determined the production of prostaglandin (PG) E2 and D2 in colon mucosa, as well as cyclooxygenase (COX)-1 and -2 and nuclear transcription factor NF-kappa B (NF-kappaB) p65 protein expression. Finally, since resveratrol has been found to modulate apoptosis, we intended to elucidate its effects on colonic mucosa under chronic inflammatory conditions. Resveratrol (10 mg kg(-1) day(-1)) significantly attenuated the damage score and corrected the disturbances in morphology associated to injury. In addition, the degree of neutrophil infiltration and the levels of TNF-alpha were significantly ameliorated. Resveratrol did not modify PGD2 levels but returned the decreased PGE2 values to basal levels and also reduced COX-2 and the NF-kappaB p65 protein expression. Furthermore, treatment of rats with resveratrol caused a significant increase of TNBS-induced apoptosis in colonic cells. In conclusion, resveratrol reduces the damage in chronic experimentally induced colitis, alleviates the oxidative events, returns PGE2 production to basal levels and stimulates apoptosis in colonic cells. PMID- 16474425 TI - IRF family proteins and type I interferon induction in dendritic cells. AB - Dendritic cells (DC), although a minor population in hematopoietic cells, produce type I interferons (IFN) and other cytokines and are essential for innate immunity. They are also potent antigen presenters and regulate adaptive immunity. Among DC subtypes plasmacytoid DC (pDC) produce the highest amounts of type I IFN. In addition, pro- and anti-inflammatory cytokines such as IL-12 and IL-10 are induced in DC in response to Toll like receptor (TLR) signaling and upon viral infection. Proteins in the IRF family control many aspects of DC activity. IRF-8 and IRF-4 are essential for DC development. They differentially control the development of four DC subsets. IRF-8-/- mice are largely devoid of pDC and CD8alpha+ DC, while IRF-4-/- mice lack CD4+DC. IRF-8-/-, IRF4-/-, double knock out mice have only few CD8a-CD4-DC that lack MHC II. IRF proteins also control type I IFN induction in DC. IRF-7, activated upon TLR signaling is required for IFN induction not only in pDC, but also in conventional DC (cDC) and non-DC cell types. IRF-3, although contributes to IFN induction in fibroblasts, is dispensable in IFN induction in DC. Our recent evidence reveals that type I IFN induction in DC is critically dependent on IRF-8, which acts in the feedback phase of IFN gene induction in DC. Type I IFN induction in pDC is mediated by MyD88 dependent signaling pathway, and differs from pathways employed in other cells, which mostly rely on TLR3 and RIG-I family proteins. Other pro inflammatory cytokines are produced in an IRF-5 dependent manner. However, IRF-5 is not required for IFN induction, suggesting the presence of separate mechanisms for induction of type I IFN and other pro-inflammatory cytokines. IFN and other cytokines produced by activated DC in turn advance DC maturation and change the phenotype and function of DC. These processes are also likely to be governed by IRF family proteins. PMID- 16474426 TI - Antiviral innate immunity pathways. AB - Recent studies have uncovered two signaling pathways that activate the host innate immunity against viral infection. One of the pathways utilizes members of the Toll-like receptor (TLR) family to detect viruses that enter the endosome through endocytosis. The TLR pathway induces interferon production through several signaling proteins that ultimately lead to the activation of the transcription factors NF-kappaB, IRF3 and IRF7. The other antiviral pathway uses the RNA helicase RIG-I as the receptor for intracellular viral double-stranded RNA. RIG-I activates NF-kappaB and IRFs through the recently identified adaptor protein MAVS, a CARD domain containing protein that resides in the mitochondrial membrane. MAVS is essential for antiviral innate immunity, but it also serves as a target of Hepatitis C virus (HCV), which employs a viral protease to cleave MAVS off the mitochondria, thereby allowing HCV to escape the host immune system. PMID- 16474427 TI - Interferon-gamma mRNA attenuates its own translation by activating PKR: a molecular basis for the therapeutic effect of interferon-beta in multiple sclerosis. AB - PKR, the interferon (IFN)-inducible protein kinase activated by double-stranded RNA, inhibits translation by phosphorylating the initiation factor eIF2alpha chain. Uniquely, human IFN-gamma mRNA uses local activation of PKR in the cell to control its own translation yield. IFN-gamma mRNA activates PKR through a structure in its 5'- region harboring a pseudoknot which is critical for PKR activation. Mutations that impair pseudoknot stability reduce the ability of IFN gamma mRNA to activate PKR and strongly increase its translation efficiency. The cis-acting RNA element in IFN-gamma mRNA functions as a biological sensor of intracellular PKR levels. During an immune response, as IFN-gamma and other inflammatory cytokines build up in the cell's microenvironment, they act to induce higher levels of PKR in the cell, resulting in a more extensive activation of PKR by IFN-gamma mRNA. With the resulting phosphorylation of eIF2alpha, a negative feedback loop is created and the production of IFN-gamma is progressively attenuated. We propose that the therapeutic effect of IFN-beta in multiple sclerosis may rest, at least in part, on its exquisite ability to induce high levels of PKR in the cell and thereby to limit IFN-gamma mRNA translation through this negative feedback loop, blocking the excessive IFN-gamma gene expression that precedes clinical attacks. PMID- 16474428 TI - Regulation of cytokine production during phagocytosis of apoptotic cells. AB - Loss of self-tolerance and expansion of auto-reactive lymphocytes are the basis for autoimmunity. Apoptosis and the rapid clearance of apoptotic cells by phagocytes usually occur as coordinated processes that ensure regulated cellularity and stress response with non-pathological outcomes. Defects in clearance of apoptotic cells would contribute to the generation of self-reactive lymphocytes, which drive autoimmune disorders such as rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). The IL-12 family of cytokines (IL-12, IL 23, and IL-27) and IL-10 are produced by phagocytic macrophages and play critical roles in the regulation of antigen-presenting cells (APCs) and effector lymphocytes during an immune response to pathogens. Inappropriate expression of these cytokines and their dysregulated activities have been strongly implicated in the pathogenesis of several autoimmune diseases. The production of pro- and anti-inflammatory cytokines by phagocytic APCs is delicately regulated during the ingestion of apoptotic cells as part of an intrinsic mechanism to prevent inflammatory autoimmune reactions. How apoptotic cell-derived signals regulate cytokine production is poorly understood. A recent study by our group demonstrated that phagocytosis of apoptotic cells by activated macrophages results in strong inhibition of IL-12 p35 gene expression by activating a novel transcription repressor, which we named GC-binding protein (GC-BP), through tyrosine dephosphorylation. We are also beginning to understand the molecular mechanisms underlying apoptotic cell-triggered production of IL-10 by phagocytes. These studies will help to elucidate some novel immune regulatory mechanisms and explore the regulation of immune responses to autoantigens with potentials to discover new therapeutic targets for the treatment of autoimmune disorders. PMID- 16474429 TI - Insight into the biology of macrophage migration inhibitory factor (MIF) revealed by the cloning of its cell surface receptor. AB - The recent cloning of MIF receptor fills an important gap in our understanding of the molecular biology and immunology of MIF. The MIF receptor, like MIF, does not fall into any established family of protein mediators, providing both new challenges and opportunities for the structural and functional analysis of MIF signal transduction. PMID- 16474430 TI - TGF-beta and cancer: is Smad3 a repressor of hTERT gene? AB - Transforming growth factor beta (TGF-beta) carries out tumor suppressor activity in epithelial and lymphoid cells, whereas telomerase is required for most cancers. Although the molecular mechanisms by which TGF-beta acts as a tumor suppressor are yet to be fully established, a link between TGFb and its tumor suppressor activity by telomerase has been suggested. Recently, we have noted a novel mode of action for TGF-beta through which human telomerase reverse transcriptase (hTERT) gene is repressed in immortal and neoplastic cells, confirming that one of the mechanisms underlying TGF-beta suppression of tumor growth may be through inhibiting hTERT gene transcription. Moreover, the inhibition of hTERT gene by TGF-beta suggests a cis action of the TGF-beta signaling molecule Smad3 on hTERT promoter directly. This article examines our current understanding and investigation of TGF-beta regulation of telomerase activity, and presents a model in which Smad3 participates in regulating hTERT gene transcription by acting as a repressor directly. Engineering the interface between Smad3 and hTERT gene may lead to a new strategy to inhibit telomerase activity in cancer. PMID- 16474431 TI - Midkine, a cytokine that inhibits HIV infection by binding to the cell surface expressed nucleolin. AB - The growth factor midkine (MK) is a cytokine that inhibits HIV infection in cell cultures in an autocrine and paracrine manner by blocking the attachment of HIV particles to permissive cells. MK mRNA is systematically expressed in adult peripheral blood lymphocytes from healthy donors, while its expression becomes markedly but transiently increased upon in vitro treatment of lymphocytes with IL 2 or IFN-gamma and activation of T lymphocytes by PHA or through the engagement of the CD28 antigen. The binding of MK to cells occurs specifically at a high and a low affinity binding site. This low affinity-binding site is the cell-surface expressed nucleolin, which is implicated in the mechanism of the initial attachment of HIV particles to cells. Accordingly, the nucleolin-binding HB-19 pseudopeptide has no effect on the MK binding to the high affinity binding site, whereas it prevents the binding of MK to the low affinity binding site, thus suggesting the low affinity receptor of MK is the cell-surface-expressed nucleolin. Confocal immunofluorescence laser microscopy revealed the colocalization of MK and the cell-surface-expressed nucleolin at distinct spots. The use of various deletion constructs of nucleolin then indicates that the extreme C-terminal end of nucleolin, containing repeats of the amino acid motif RGG, as the domain that binds MK. The specific binding of MK to the surface nucleolin is independent of heparan sulfate and chondroitin sulfate proteoglycans. After binding to cells, MK enters cells by an active process in which nucleolin and lipid rafts appear to be implicated. The potent and the distinct anti-HIV action of MK along with its enhanced expression in lymphocytes by various physiological stimuli, point out that MK is a cytokine that could be involved in HIV pathogenesis. PMID- 16474432 TI - Therapy of cancer by cytokines mediated by gene therapy approach. AB - Gene therapy offers a new approach for treatment of cancer. Transfer of genes encoding immunostimulatory cytokines has been used with remarkable success to eliminate cancer in animals. However, clinical trials in patients with this strategy had limited efficacy. Therefore, improvement of gene transfer vector system is necessary. A hybrid viral vector, consisting of SFV replicon with either murine IL-12 or reporter LacZ gene, was constructed. This hybrid vector showed specificity and high level of expression in HCC both in vitro and in vivo. In a rat orthotropic liver tumor model, treatment of established tumors by the hybrid vector with mIL-12 gene resulted in a strong anti-tumor activity without accompanying toxicity. Subsequently, a helper-dependent adenovirus vectors containing a mifepristone (RU486) inducible system was constructed for controlled and liver-specific expression of human interleukin 12 (hIL-12) (HD-Ad/RUhIL-12) and mouse IL-12 (mIL-12) (HD-Ad/RUmIL-12). Data showed that high and sustained serum levels of hIL-12 could be attained by continuing administration of RU486 every 12 or 24 h. Repetitive induction of hIL-12 could be obtained over, at least, a period of 48 weeks after a single injection of HD-Ad/RUhIL-12. Treatment of liver metastases with of HD-Ad/RUmIL-12 plus RU846 resulted in complete tumor regression in all animals. Then, different cytokine genes were inserted into conditional replicative adenoviruses vectors (also called oncolytic adenovirus). Replication of adenovirus in tumor cells would kill tumor cells and release viruses, which infect surrounding tumor cells. The combination of cytopathic effect by oncolytic adenovirus and biological effect of transgene would exert strong antitumor activity. These new types of vectors may provide a potent and safe tool for cancer gene therapy. PMID- 16474433 TI - Local signals in stem cell-based bone marrow regeneration. AB - The cellular basis of bone marrow (BM) tissue development and regeneration is mediated through hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs). Local interplays between hematopoietic cells and BM stromal cells (BMSCs) determine the reconstitution of hematopoiesis after myelosuppression. Here we review the BM local signals in control of BM regeneration after insults. Hematopoietic growth factors (HGFs) and cytokines produced by BMSCs are primary factors in regulation of BM hematopoiesis. Morphogens which are critical to early embryo development in multiple species have been added to the family of HSCs regulators, including families of Wnt proteins, Notch ligands, BMPs, and Hedgehogs. Global gene expression analysis of HSCs and BMSCs has begun to reveal signature groups of genes for both cell types. More importantly, analysis of global gene expression coupled with biochemical and biological studies of local signals during BM regeneration have strongly suggested that HGFs and cytokines may not be the primary local regulators for BM recovery, rather chemokines (SDF 1, FGF-4) and angiogenic growth factors (VEGF-A, Ang-1) play instructive roles in BM reconstitution after myelosuppression. A new direction of management of BM toxicity is emerging from the identification of BM regenerative regulators. PMID- 16474434 TI - Regulation of cytokine signaling pathways by PIAS proteins. AB - Cytokines activate multiple signal transduction pathways to regulate gene expression. STATs and NF-kB are two important families of transcription factors activated by cytokines. Abnormal regulation of STAT and NF-kB activities has been associated with human diseases. The protein inhibitor of activated STAT (PIAS) protein family has been proposed to interact with over 60 proteins, many of which are transcription factors involved in the immune system. PIAS proteins regulate transcription through several mechanisms, including blocking the DNA-binding activity of transcription factors, recruiting transcriptional co-repressors and promoting protein sumoylation. This article is to review the role of PIAS proteins in the regulation of STAT and NF-kB signaling pathways. PMID- 16474435 TI - How the Bcl-2 family of proteins interact to regulate apoptosis. AB - Commitment of cells to apoptosis is governed largely by protein-protein interactions between members of the Bcl-2 protein family. Its three sub-families have distinct roles: the BH3-only proteins trigger apoptosis by binding via their BH3 domain to pro-survival relatives, while the pro-apoptotic Bax and Bak have an essential downstream role involving disruption of organellar membranes and induction of caspase activation. The BH3-only proteins act as damage sensors, held inert until their activation by stress signals. Once activated, they were thought to bind promiscuously to pro-survival protein targets but unexpected selectivity has recently emerged from analysis of their interactions. Some BH3 only proteins also bind to Bax and Bak. Whether Bax and Bak are activated directly by these BH3-only proteins, or indirectly as a consequence of BH3-only proteins neutralizing their pro-survival targets is the subject of intense debate. Regardless of this, a detailed understanding of the interactions between family members, which are often selective, has notable implications for designing anti-cancer drugs to target the Bcl-2 family. PMID- 16474436 TI - STAT3 in immune responses and inflammatory bowel diseases. AB - STAT3 has been known as a mediator for gene expression induced by many important cytokines. Recent studies have suggested that STAT3 has important functions in regulation of both innate and adaptive immunity. Loss of STAT3 in immune cells caused severe inflammation in response to pathogens. This review discusses the recent progress and suggests directions for the future research on this interesting molecule. PMID- 16474437 TI - Characterization of the antiviral effects of interferon-alpha against a SARS-like coronoavirus infection in vitro. AB - Interferon (IFN)-alphas bind to and activate their cognate cell surface receptor to invoke an antiviral response in target cells. Well-described receptor-mediated signaling events result in transcriptional regulation of IFN sensitive genes, effectors of this antiviral response. Results from a pilot study to evaluate the clinical efficacy of IFN-alpha treatment of SARS patients provided evidence for IFN-inducible resolution of disease. In this report we examined the contribution of IFN-inducible phosphorylation-activation of specific signaling effectors to protection from infection by a SARS-related murine coronavirus, MHV-1. As anticipated, the earliest receptor-activation event, Jak1 phosphorylation, is critical for IFN-inducible protection from MHV-1 infection. Additionally, we provide evidence for the contribution of two kinases, the MAP kinase p38MAPK, and protein kinase C (PKC) delta to antiviral protection from MHV-1 infection. Notably, our data suggest that MHV-1 infection, as for the Urbani SARS coronoavirus, inhibits an IFN response, inferred from the lack of activation of pkr and 2'5'-oas, genes associated with mediating the antiviral activities of IFN alphas. To identify potential target genes that are activated downstream of the IFN-inducible signaling effectors we identified, and that mediate protection from coronavirus infection, we examined the gene expression profiles in the peripheral blood mononuclear cells of SARS patients who received IFN treatment. A subset of differentially regulated genes were distinguished with functional properties associated with antimicrobial activities. PMID- 16474438 TI - Redox regulation of mast cell histamine release in thioredoxin-1 (TRX) transgenic mice. AB - Thioredoxin-1 (TRX) is a stress-inducible redox-regulatory protein with antioxidative and anti-inflammatory effects. Here we show that the release of histamine from mast cells elicited by cross-linking of high-affinity receptor for IgE (FcepsilonRI) was significantly suppressed in TRX transgenic (TRX-tg) mice compared to wild type (WT) mice. Intracellular reactive oxygen species (ROS) of mast cells stimulated by IgE and antigen was also reduced in TRX-tg mice compared to WT mice. Whereas there was no difference in the production of cytokines (IL-6 and TNF-alpha) from mast cells in response to 2,4-dinitrophenylated bovine serum albumin (DNP-BSA) stimulation in TRX-tg and WT mice. Immunological status of TRX tg mice inclined to T helper (Th) 2 dominant in primary immune response, although there was no difference in the population of dendritic cells (DCs) and regulatory T cells. We conclude that the histamine release from mast cells in TRX-tg mice is suppressed by inhibition of ROS generation. As ROS are involved in mast cell activation and facilitate mediator release, TRX may be a key signaling molecule regulating the early events in the IgE signaling in mast cells and the allergic inflammation. PMID- 16474440 TI - Treatment of chronic prostatitis. PMID- 16474441 TI - Early cystectomy for clinical stage T1 bladder cancer. PMID- 16474463 TI - Surgical intervention in patients with metastatic renal cancer: current status of metastasectomy and cytoreductive nephrectomy. AB - It is estimated that in 2004 there will be 35,700 new cases of, and 12,480 deaths from, kidney cancer in the US. Since 1950 there has been a 126% increase in the incidence of renal cancer and a 36.5% increase in annual associated mortality. In the past two decades, our understanding of tumors arising from the renal cortex has dramatically expanded owing to advances in cytogenetics and histopathological reclassification. It is now known that renal cell carcinoma (RCC) is a family of neoplasms that possess unique molecular and cytogenetic defects, with 90% of metastases emanating from conventional clear cell carcinoma subtype. In addition to advancing our understanding of RCC, improved abdominal imaging technology has caused a migration of tumor stage and alteration of surgical strategies, with tumors commonly being diagnosed at an earlier stage. Despite these advances, the prognosis for patients with metastatic RCC is poor. Studies that examine combinations of surgery and systemic therapy aim to improve survival in this high risk group. PMID- 16474464 TI - Botulinum toxin in urology: evaluation using an evidence-based medicine approach. AB - Clinical reports documenting the beneficial effects of botulinum toxin (BTX) to treat various lower urinary tract disorders of muscle spasticity (e.g. detrusor hyperreflexia, detrusor sphincter dyssynergia) have led to its expanded application in non-neurogenic conditions such as idiopathic detrusor overactivity, detrusor underactivity and benign prostatic hyperplasia (BPH). As greater numbers of investigators report their results utilizing this agent, it becomes more important that clinicians are able to synthesize and describe the findings of current clinical studies in a meaningful way. Evidence-based medicine is a platform on which investigations concerning a particular topic can be evaluated for their levels of scientific evidence, so that rational clinical recommendations can be formulated. This review article applies evidence-based practices to the use of BTX in common urologic conditions. Appreciating the benefits as well as limitations of currently available studies will hopefully drive the development of well-controlled, randomized studies of BTX, particularly in highly prevalent conditions such as idiopathic detrusor overactivity and BPH. PMID- 16474465 TI - Current views on evaluation, management, and gender assignment of the intersex infant. AB - In recent years, traditional views regarding the management of infants with intersex conditions have been challenged. Some of these patients have gender dysphoria and gender conversion, although most adults with intersex consider themselves to be either male or female. Hormonal and genetic factors may have a more important role in gender identity and sexual satisfaction than previously recognized, whereas the importance of phallus size to male gender identity and sexual satisfaction may have been overestimated. The impact of androgen imprinting on the developing brain is uncertain, but it is likely to be significant. The issue of genital surgery in infancy is controversial, although many adult patients concur that infancy is the best time for such procedures. Several reports indicate that the functional outcomes of genital surgery are poor, although more recently developed surgical techniques may achieve better results. Good communication between physicians, patients, and families regarding intersex conditions is paramount. We review current understanding of the evaluation and management of intersex conditions. PMID- 16474466 TI - Technology insight: Novel ureteral stent materials and designs. AB - Ureteral stents are an important tool for aiding upper urinary tract drainage, but can cause significant patient morbidity. Common problems include stent induced pain, hematuria, dysuria, infection, and encrustation. From a urologist's perspective, stents must be easy to maneuver in the urinary tract, radiopaque, and affordable. Since the development of the modern day stent in 1978, stents have evolved to include softer biomaterials that are more resistant to encrustation and infection. An ideal biomaterial is one that is not affected by its environment and does not elicit reactive changes in surrounding tissues. To date, the ideal biomaterial or stent does not exist. This review discusses developments that address the issues of infection, biofilm formation, encrustation, and patient comfort. Stent materials including polyurethane, silicone, biodegradable substances and new combination polymers are reviewed, in addition to novel stent coatings such as heparin, hydrogel, and silver nitrate. Ureteral stent technologies currently lag behind vascular stents, particularly drug-eluting stents, but new developments will continue to improve these essential urological tools. PMID- 16474467 TI - Erectile dysfunction and priapism. AB - BACKGROUND: A 46-year-old man presented with erectile dysfunction following a blunt perineal injury sustained 1 year previously. Oral phosphodiesterase type 5 inhibitors failed to improve his erections. The remainder of the patient's history, physical examination and laboratory tests were unremarkable except for a moderate tenderness in the left perineum and some 'fullness' in the corpora cavernosa during physical examination. INVESTIGATION: Physical examination, color duplex ultrasound of the penis and perineal area and Sexual Health Index for Men questionnaire. DIAGNOSIS: Non-ischemic priapism due to a ruptured left helicine artery, intermittent penile turgidity and erectile dysfunction. MANAGEMENT: Open suture ligation of the helicine artery and imbrication of the pseudocapsule. PMID- 16474468 TI - Content coverage in Nature Clinical Practice Urology: a balancing act. PMID- 16474469 TI - Open partial nephrectomy: an essential contemporary operation. PMID- 16474470 TI - Prostate cancer: risks and benefits of screening. PMID- 16474491 TI - Prophylactic use of antimicrobials in commonly performed outpatient urologic procedures. AB - An antimicrobial is an agent capable of killing or inhibiting the growth of a micro-organism. Antimicrobial prophylaxis encompasses efforts to prevent postprocedure infections through the use of an antimicrobial agent before, and, in some cases, for a limited time after a procedure. A thorough history and physical examination are essential to identify host factors that increase a patient's risk for postprocedural infection. Risk-modifying factors include age, anatomy, geographical area of residence, immune and nutritional status, cardiac valve integrity, prosthetic joints, the presence of indwelling hardware and distant infectious wounds. Prophylaxis for the most common urologic outpatient procedures can be attained with oral trimethoprim-sulfamethoxazole or fluoroquinolone administered between 2 h and 30 min before a procedure. Special consideration for the type and duration of prophylaxis should be given to patients with moderate to severe cardiac valvular conditions and recently inserted prosthetic joints. PMID- 16474493 TI - Urodynamic assessment of bladder-outlet obstruction in women. AB - Bladder-outlet obstruction (BOO) in women has become an increasingly recognized entity over the past several years. This is partly because of the greater number of anti-incontinence surgeries being performed, which can infrequently result in iatrogenic obstruction; and partly because of improvements in the ability to diagnose and recognize BOO. In this review, the symptoms associated with BOO in women, and whether these symptoms can reliably predict the presence of BOO, will be discussed. The incidence of BOO following surgical anti-incontinence procedures and the urodynamic diagnosis of BOO in women, detailing the various criteria that have been proposed, are also reviewed. PMID- 16474492 TI - Indications and contraindications for the use of laparoscopic surgery for renal cell carcinoma. AB - Surgery remains the only treatment with a chance of cure for renal cell carcinoma. Laparoscopic radical nephrectomy (LRN) has developed to be a standard treatment for the management of suspected renal malignancy in many centers worldwide, with oncologic efficacy equal to that of open radical nephrectomy. LRN has considerable advantages over open surgery, such as decreased postoperative morbidity, decreased analgesic requirements, and shorter hospital stay and convalescence. Current indications for LRN include all patients with localized stage T1-2 renal tumors. LRN for stage T3 renal tumors may be technically feasible in individual situations, but cannot be considered standard treatment. Open radical nephrectomy is reserved for advanced renal tumors, according to the surgeon's judgment. Partial nephrectomy is well established and considered to be the standard management for all organ-confined tumors of 2 cm) and multiple strictures, untreated perioperative urinary infection and extensive periurethral spongiofibrosis. Repeated IU might be useful in patients who have a stricture recurrence more than 6 months after the initial procedure, but repeat IU offers no long-term cure after a third IU, or if a stricture recurs within 3 months of the first IU. Such patients should be offered urethroplasty. Repeated IU followed by long-term self-dilation is an alternative option for men with severe comorbidity and limited life expectancy, or those who have failed previous urethroplasty. Overall, IU has a lower success rate (+/-60%) than urethroplasty (+/-80-90%), but if used for selected strictures, the success rate of IU could approach that of urethroplasty. PMID- 16474598 TI - Mechanisms of Disease: specialized interstitial cells of the urinary tract--an assessment of current knowledge. AB - Scientists interested in the smooth muscles of the urinary tract, and their control, have recently been studying cells in the interstitium of tissues that express the c-kit antigen (Kit(+) cells). These cells have morphologic features that are reminiscent of the well-described pacemaker cells in the gut, the interstitial cells of Cajal (ICC). The spontaneous contractile behavior of muscles in the urinary tract varies widely, and it is clear that urinary tract Kit(+) interstitial cells cannot be playing an identical role to that played by the ICC in the gut. Nevertheless, there is increasing evidence that they do play a role in modulating the contractile behavior of adjacent smooth muscle, and might also be involved in mediating neural control. This review outlines the properties of ICC in the gut, and gives an account of the discovery of cells in the interstitium of the main components of the urinary tract. The physiologic properties of such cells and the functional implications of their presence are discussed, with particular reference to the bladder. In this organ, Kit(+) cells are found under the lamina propria, where they might interact with the urothelium and with sensory nerves, and also between and within the smooth-muscle bundles. Confocal microscopy and calcium imaging are being used to assess the physiology of ICC and their interactions with smooth muscles. Differences in the numbers of ICC are seen in smooth muscle specimens obtained from patients with various pathologies; in particular, bladder overactivity is associated with increased numbers of these cells. PMID- 16474599 TI - Primer: questionnaires versus urodynamics in the evaluation of lower urinary tract dysfunction-one, both or none? AB - What is the role of urodynamic assessments and have we made too much use of them? Is there a correlation between symptoms and underlying pathophysiology? These questions are addressed in this article. There are disparities in the assessment of lower urinary tract dysfunction between the accuracy of symptomatic assessment and the underlying pathophysiology. This is particularly evident with voiding symptoms, in contrast to storage symptoms, and has fuelled the debates that have resulted in the abandonment of the term 'prostatism' in favor of 'lower urinary tract symptoms', whereas the term 'overactive bladder syndrome' has been embraced as a storage symptom complex. Clearly, voiding disorders principally affect men, whilst storage disorders are more common in women. Much has been written and spoken about regarding the use of symptomatic assessment versus urodynamics in the assessment of lower urinary tract symptoms in men, whilst there has been a clearer consensus on the potential usefulness of urodynamics in the assessment of storage disorders, and so this article focuses principally on the latter topic. PMID- 16474600 TI - The importance of MRI evaluation in the preoperative work-up of prostate cancer. AB - BACKGROUND: A 59-year-old man on exogenous androgen therapy presented with a clinically palpable prostate nodule confined to one lobe on endorectal examination. Serum prostate-specific antigen was 3.4 ng/ml. Bone scintigraphy and baseline CT were reportedly negative. INVESTIGATIONS: Physical examination, laboratory evaluation, endorectal MRI of the prostate, cystoscopy and biopsy. DIAGNOSIS: Poorly differentiated adenocarcinoma of the prostate, with invasion of the urinary bladder. MANAGEMENT: Neoadjuvant hormonal therapy, followed by radiation. PMID- 16474601 TI - Regionalization of care: centralizing complex surgical procedures. PMID- 16474602 TI - Nomograms for prostate cancer--is their use evidence-based? PMID- 16474623 TI - Therapy Insight: bladder dysfunction associated with multiple sclerosis. AB - Bladder dysfunction is a common problem for patients with multiple sclerosis. The severity of symptoms often correlate with the degree of spinal cord involvement and, hence, the patient's general level of disability. The emphasis of management is now mainly medical and is increasingly offered by nonurologists. Treatments can be highly effective, relieving patients of what are otherwise very troublesome symptoms that would compound their neurological disability. This article gives an overview of the neural control of the bladder, followed by an explanation of the pathophysiology of detrusor overactivity secondary to neurological disease. A review of methods available for treating bladder dysfunction in multiple sclerosis then follows. The treatment options for this disorder are largely medical and include established first-line measures such as anticholinergics, clean intermittent self-catheterization and the use of desmopressin, as well as potential second-line agents, such as cannabinoids, intravesical vanilloids and intradetrusor botulinum neurotoxin type A. The diminishing role of surgical intervention is also discussed. PMID- 16474622 TI - Prostate-specific antigen doubling time as a prognostic marker in prostate cancer. AB - Prostate cancer has a varied natural history. Men with similar serum prostate specific antigen (PSA) levels, clinical stages, and histologic features in their tissue specimens can have markedly different outcomes. While prostate cancer is lethal in some patients, most men die with cancer rather than because of it. Moreover, histologically apparent cancer can be found in the prostate glands of approximately 42% of men over 50 years of age who die from other causes, but the lifetime risk that a man in the US will be diagnosed with prostate cancer is estimated to be 11% and the risk of dying from the disease is only 3.1%. Consequently, appropriate disease management requires risk assessment. How likely is it that a given man's cancer will progress or metastasize over his remaining lifetime? What is the probability of successful treatment? What are the risks of adverse effects and complications of each treatment? Physicians use a variety of clinical and pathologic parameters to assess the risk that a given cancer poses to an individual patient. In addition to the accepted parameters of serum PSA level, clinical staging, and pathologic grading and staging, PSA doubling time has emerged as an important factor in the evaluation of men with newly diagnosed prostate cancer or prostate cancer that recurs after treatment. PSA doubling time can also be used as a surrogate marker for prostate cancer-specific death. This review summarizes current knowledge regarding the role of PSA doubling time as a prognostic marker in men with prostate cancer. PMID- 16474624 TI - Mechanisms of Disease: genetic and epigenetic alterations that drive bladder cancer. AB - There is substantial evidence for the existence of mutually exclusive molecular pathways to tumorigenesis, in the formation of papillary and invasive carcinomas, respectively. The most common genetic alterations in low grade papillary transitional-cell carcinoma (TCC) are loss of heterozygosity of part or all of chromosome 9 and activating mutations of the fibroblast growth factor receptor 3 (FGFR3). The pathway to development of invasive TCC seems to start with dysplasia, progress to carcinoma in situ, followed by invasion of the lamina propria. The most frequent genetic alteration in dysplasia and carcinoma in situ is mutation of TP53, followed by loss of heterozygosity of chromosome 9. A marker for progression in TCC is loss of chromosome 8p, which occurs in approximately 60% of bladder tumors. Global trends of increased genomic instability and aberrant methylation of cytosine residues in DNA correlate with increased tumor invasion and progression. When researching markers of bladder cancer for clinical use, it is important that biomedical pathways and their alterations are measured in the same tumor populations. This review examines the published data and proposes a model for the mechanisms behind bladder cancer development. PMID- 16474625 TI - Ureterosigmoidostomy and obstructive uropathy. AB - BACKGROUND: A 19-year-old mentally retarded man with failed exstrophy repair and ureterosigmoidostomy urinary diversion presented with high fever, vomiting and right-flank pain of 2 days' duration. Past medical history was notable for a left nephrectomy to treat an infected staghorn calculus in a poorly functioning kidney. Physical examination revealed pyrexia and right-flank tenderness. INVESTIGATIONS: Physical examination, renal function tests, electrolyte and metabolic assessment, urine and blood cultures, abdominal CT, ANTEGRADE PYELOURETEROGRAPHY, sigmoidoscopy and histopathology. DIAGNOSIS: Ureterosigmoidostomy complicated by acute pyelonephritis, obstructive uropathy, recurrent urinary tract infections, renal impairment and the development of renal stones and metabolic acidosis. MANAGEMENT: Fluids, intravenous antibiotics, bicarbonate and potassium supplementation, and rediversion of ureterosigmoidostomy to an ileal conduit. PMID- 16474626 TI - Molecular medicine and its impact on practice. PMID- 16474627 TI - Laparoscopic versus robotic radical prostatectomy. PMID- 16474653 TI - Suitability of different sling materials for the treatment of female stress urinary incontinence. AB - Stress urinary incontinence (SUI) is defined as leakage of urine with a sudden increase in intra-abdominal pressure, such as that seen with laughing, lifting, or changing position, without a concomitant rise in detrusor (bladder-generated) pressure. The proposed mechanism of SUI is that an increase in intra-abdominal pressure resulting from various activities causes the bladder pressure to rise above the urethral pressure. The pubovaginal sling remains the standard treatment for female SUI in the US. The market has been flooded with innumerable sling materials. This review discusses the currently available sling materials, surgical approaches, and clinical outcomes data. Long-term data on efficacy is lacking, but early results with new materials and delivery techniques indicate that excellent cure rates with minimal morbidity and high patient satisfaction may be achievable. PMID- 16474655 TI - Mechanisms of disease: Polymorphisms of androgen regulatory genes in the development of prostate cancer. AB - Androgens are of primary importance in the etiology of prostate cancer, and binding of the androgen dihydrotestosterone to the androgen receptor is thought to stimulate prostate growth. It has been proposed that polymorphisms within key androgen regulatory genes may contribute to an individual's risk of developing prostate cancer. Attributing single polymorphisms to complex, late-onset, chronic diseases such as prostate cancer is probably not feasible, but identification of genes that increase risk will contribute to larger-scale multigenic risk assessment. Here, we review the current status of our knowledge of associations between important androgen regulatory gene polymorphisms and prostate cancer risk. PMID- 16474654 TI - Drug insight: Use of docetaxel in prostate and urothelial cancers. AB - Taxanes have emerged as a potent class of chemotherapeutic agents in many malignancies, with two taxanes now in clinical use. Their mechanism of action against tumor cells is by alteration of microtubule dynamics, which causes cell cycle arrest during mitosis. Docetaxel binds to the microtubules with a higher affinity than paclitaxel, and over a broader range of cell-cycle activities. It has also been shown to promote apoptosis via BCL2 phosphorylation. In hormone refractory prostate cancer, docetaxel has been studied as both a single agent and in combination with estramustine, and in different treatment schedules, with demonstrated efficacy. Two phase III trials have confirmed a survival benefit, making docetaxel the first chemotherapy agent with proven efficacy against prostate cancer. In urothelial cancer, docetaxel has demonstrated activity and has been investigated as a single agent and in combination regimens. A phase III trial comparing docetaxel and cisplatin to methotrexate, vinblastine, doxorubicin, and cisplatin was inferior when evaluating response rates and overall survival. More recent phase II trials combining docetaxel with two additional agents have shown promise, but confirmatory trials are needed. PMID- 16474657 TI - New technology and the changing world of cancer. PMID- 16474656 TI - Emphysematous pyelonephritis. AB - BACKGROUND: A 54-year-old nondiabetic male presented with high fever, vague lower abdominal pain and leakage of urine around his long-standing suprapubic catheter. Examination revealed pyrexia and tenderness in the right renal angle. White cell count was 22.8 x 10(9)/l. Plain abdominal X-ray showed calculi in the right kidney, ureter and bladder. Intravenous pyelogram showed gas confined to the right upper renal pelvis and perinephric space. INVESTIGATIONS: Urine and blood cultures, plain abdominal X-ray, intravenous pyelogram, abdominal ultrasound, MAG3 renogram and histopathology. DIAGNOSIS: Emphysematous pyelonephritis: class 2 or type 1. Escherichia coli was isolated from urine obtained by endoscopic drainage. MANAGEMENT: Endoscopic drainage of pus and simple nephrectomy. PMID- 16474658 TI - Interpreting data from randomized trials: the Scandinavian prostatectomy study illustrates two common errors. PMID- 16474680 TI - The etiology and management of premature ejaculation. AB - Premature ejaculation (PE) is a common male sexual disorder. Normative data suggest that men with an intravaginal ejaculatory latency time of less than 1 min have 'definite' premature ejaculation, while men with intravaginal ejaculatory latency times of between 1 and 1.5 min have 'probable' premature ejaculation. Although there is insufficient empirical evidence to identify the etiology of PE, there is correlational evidence to suggest that men with PE have high levels of sexual anxiety and altered sensitivity of central 5-hydroxytryptamine receptors. Pharmacological modulation of the ejaculatory threshold using daily or on-demand selective serotonin reuptake inhibitors offers patients a high likelihood of achieving improved ejaculatory control within a few days of initiating treatment, leads to improvements in sexual desire and other sexual domains, and is well tolerated. PMID- 16474682 TI - From coagulation to enucleation: the use of lasers in surgery for benign prostatic hyperplasia. AB - The application of lasers for the treatment of benign prostatic hyperplasia has evolved over the past 15 years. Early-generation neodymium:yttrium aluminum garnet lasers were used to coagulate and ablate prostatic tissue, but significant postoperative irritative symptoms and high reoperation rates meant that this approach did not seriously challenge the status quo for long. Ablative techniques have recently become popular again with the marketing of the newer-generation, higher-power potassium titanyl phosphate and holmium lasers. Although short-term data are encouraging, there are no comparative trials of significant duration, so it is not yet possible to draw conclusions with regard to efficacy and durability. The holmium laser can also be used as an incisional and dissecting tool that allows resection or enucleation of whole lobes of the prostate, mimicking the action of the index finger in open prostatectomy. The safety, efficacy, durability and cost-effectiveness of the holmium laser have been shown in well-designed randomized controlled trials. PMID- 16474681 TI - Magnetic resonance imaging and magnetic resonance spectroscopic imaging of prostate cancer. AB - Magnetic resonance imaging (MRI) and magnetic resonance spectroscopic imaging (MRSI) are evolving techniques that offer noninvasive evaluation of anatomic and metabolic features of prostate cancer. The ability of MRI to determine the location and extent of the tumor and to identify metastatic spread is useful in the pretreatment setting, enabling treatment decision-making that is evidence based. MRSI of the prostate gland expands the diagnostic assessment of prostate cancer through the detection of cellular metabolites, and can lead to noninvasive differentiation of cancer from healthy tissue. MRI/MRSI can also be used to evaluate both local and systemic recurrence, with endorectal MRI being capable of detecting local recurrence, even in patients with rising serum PSA level but no palpable tumor on digital rectal examination. Considering the benefits that MRI and MRSI have been shown to offer patients, the skills and technology required to perform these tests should be widely disseminated to make their routine use possible. Teamwork between members of radiology, pathology, urology and radiation oncology departments is essential in order to exploit these technologies fully. PMID- 16474683 TI - Therapy insight: Priapism associated with hematologic dyscrasias. AB - Priapism is defined as a genuine erectile disorder, in which erection persists uncontrollably without sexual purpose. This disorder is often overlooked in comparison to other male sexual dysfunctions, principally because of its seeming rarity. Priapism, however, occurs frequently in certain patient populations, including those with hematologic dyscrasias. Most commonly, priapism affects patients with sickle cell disease. The reported prevalence rate in this group ranges between 29% and 42% of males. The significance of priapism is further emphasized by the fact that this disorder commonly causes erectile tissue damage and leads to the loss of functional erections. These observations highlight the need for proper clinical recognition of priapism in order to enable appropriate management. Here, the current knowledge of priapism as it relates to hematologic and coagulative diseases is discussed, with a focus on the diagnosis and treatment of the disorder. PMID- 16474684 TI - Bilateral metachronous testicular seminoma. AB - BACKGROUND: A 33-year-old male presented to hospital with a painless right testicular mass measuring 2 x 3 cm and no systemic symptoms. The patient had no relevant medical history. CT and blood tests did not reveal metastasis and the patient was treated with a right inguinal orchiectomy and radiotherapy. After 4 years a routine follow-up examination revealed a 1 cm palpable mass in the left testis. INVESTIGATIONS: CT, tumor marker assessment, scrotal ultrasonography, Doppler ultrasound examination, and biopsy. DIAGNOSIS: Bilateral metachronous testicular seminoma, stage pT1N0M0S0. MANAGEMENT: Radiotherapy, radical orchiectomy, bilateral placement of testicular prostheses, androgen substitution therapy, regular follow-up assessment. PMID- 16474685 TI - The physician and conflict of interest. PMID- 16474686 TI - Penile enlargement surgery--fact or illusion? PMID- 16474710 TI - Active surveillance with selective delayed intervention is the way to manage 'good-risk' prostate cancer. AB - This review summarizes the case for active surveillance of 'good-risk' prostate cancer, with selective delayed intervention for rapid biochemical progression, assessed by rising prostate-specific antigen (PSA) levels or grade progression. The results of a large phase II trial using this approach are also reviewed. A prospective phase II study of active surveillance with selective delayed intervention was initiated in 1995. Patients were managed initially with surveillance; those who had a PSA doubling time (PSADT) of < or = 2 years, or grade progression on repeat biopsy, were offered radical intervention. The remaining patients were closely monitored. The cohort now consists of 299 patients with good-risk--or, in men over 70 years of age, intermediate-risk- prostate cancer. The median PSADT was 7 years, 42% had a PSADT > 10 years. The majority of patients remain on surveillance. At 8 years, overall actuarial survival was 85%, and disease-specific survival was 99%. To date, this study has shown that most men with 'good-risk' prostate cancer will die of unrelated causes. The approach of active surveillance with selective delayed intervention based on PSADT represents a practical compromise between radical therapy for all patients, which results in overtreatment for patients with indolent disease, and watchful waiting with palliative therapy only, which results in undertreatment for those with aggressive disease. The results at 8 years were favorable. Longer follow-up will be required if the study is to confirm the safety of this approach in men with a long life expectancy (> 15 years). PMID- 16474712 TI - How close are we to preventing prostate cancer? PMID- 16474711 TI - Technology insight: Applications of tissue engineering and biological substitutes in urology. AB - Patients suffering from diseased or injured organs may be treated with transplanted organs. However, there is a severe shortage of donor organs, which is worsening yearly owing to the ageing population. Scientists in the field of regenerative medicine and tissue engineering apply the principles of cell transplantation, materials science, and bioengineering to construct biological substitutes that will restore and maintain normal function in diseased and injured tissues. This article reviews recent advances in regenerative medicine and describes applications of biological substitutes that may offer novel therapies for patients with end-stage organ failure. PMID- 16474713 TI - The evaluation and staging of clinically localized prostate cancer. PMID- 16474733 TI - Is histologic subtype a significant prognostic factor in renal cell carcinoma? PMID- 16474734 TI - Endourologic treatment of upper urinary tract transitional cell carcinoma. AB - The traditional treatment for upper tract transitional cell carcinoma (UTTCC) consists of radical nephroureterectomy. A more conservative approach, however, was required in cases of bilateral UTTCC and in patients with disease in a solitary kidney but who had underlying comorbidities that made them unsuitable candidates for open surgery. Minimally invasive treatment methods were developed for these select groups of patients. Because of technological advancements and refinement in endoscopic techniques, most patients with UTTCC, even those with normal contralateral kidneys, can now be offered minimally invasive treatment with single or multimodal approaches involving ureteroscopy or percutaneous resection. For patients with low-stage, low-grade UTTCC, five-year survival rates are comparable for those treated endourologically and those treated by nephroureterectomy. High-grade lesions have much higher recurrence and progression rates than lower-grade lesions, and nephroureterectomy is therefore recommended in patients with high-grade disease. The use of adjuvant instillation in the treatment of UTTCC, administered via antegrade and retrograde methods, has been shown to improve outcomes. For recurrences to be diagnosed and treated in a timely manner, and acceptable cancer-free survival rates maintained, long-term rigorous follow-up after endourologic treatment, with regular surveillance ureteroscopy, is crucial. PMID- 16474735 TI - The natural history of small renal masses. AB - The incidence of renal cell carcinoma is increasing, in part due to the growing use of cross-sectional imaging. Most renal tumors are now incidentally detected as small masses in asymptomatic patients. A minority of small renal masses, presumed to be renal cell carcinoma, grow significantly over time if managed conservatively, but the growth rate of the majority is slow or undetectable. In the absence of other prognostic factors, measurement of tumor growth rate can be helpful for initial conservative management of selected patients with small renal tumors. To date, there have been no reports of progression to metastatic disease occurring during active surveillance, but longer follow-up is needed to confirm this observation. The standard of care for small localized renal neoplasms is partial or radical nephrectomy. At the present time, active surveillance of small renal masses, with delayed therapy for patients whose disease progresses, is an experimental approach that can be considered for the elderly or patients with significant comorbidity. Renal core biopsy and fine-needle aspiration can provide essential information for treatment decision-making and should therefore be considered in the diagnostic work-up of all small renal masses. In future, the identification of prognostic indicators, with the use of new techniques including functional imaging and molecular or genomic characterization of tissue from needle biopsies, are expected to help clinicians differentiate between indolent and potentially aggressive small renal tumors. PMID- 16474736 TI - Reconstructive surgery in penile trauma and cancer. AB - This article provides an overview of the current concepts in reconstructive surgery following penile trauma, penile fracture and penile cancer. It covers the initial management of penile trauma, with the aim of preservation of as much viable tissue as is practical, and also provides advice on dealing with penile avulsion and amputation injuries. The best treatment for penile fracture immediate surgical exploration and repair-is outlined and discussed. Finally, penile cancer management is reviewed, from initial biopsy to definitive treatment of the penile lesion-including wide excision, partial glansectomy, total glansectomy, and partial and total penectomy. It is concluded that appropriate surgery in all these conditions reduces subsequent long-term problems in sexual function, cosmesis, psychology, and (in cancer cases) longevity. The same reconstructive techniques can be applied for different penile conditions, and it is suggested that surgeons become experienced in genital surgery as a whole, rather than in oncology or trauma alone. PMID- 16474737 TI - Adult paratesticular rhabdomyosarcoma. AB - BACKGROUND: A 29-year-old male patient presented with a history of painless enlargement of the left hemiscrotum. INVESTIGATIONS: Laboratory tests for beta human chorionic gonadotrophin, alpha-fetoprotein, and lactate dehydrogenase, physical examination, and CT of the chest, abdomen and pelvis. Histologic examination, nerve-sparing retroperitoneal lymph-node dissection. DIAGNOSIS: Paratesticular rhabdomyosarcoma with lymph node metastasis. MANAGEMENT: Inguinal radical orchiectomy and adjuvant chemotherapy for 48 weeks. Radiotherapy and additional chemotherapy were administered following local recurrence. PMID- 16474738 TI - Research training in urology. PMID- 16474739 TI - Which pediatric urological conditions can be handled by adult urologists? PMID- 16474760 TI - Rising prostate-specific antigen after primary prostate cancer therapy. AB - An estimated 20-40% of men experience a biochemical recurrence within 10 years of definitive prostate cancer treatment. No single prostate-specific antigen (PSA) value is invariably associated with clinical metastasis or cancer-specific survival; PSA kinetics might prove to be a more important predictor of eventual progression-free survival and cancer-specific survival than absolute PSA level alone. With only one-third of patients progressing from biochemical recurrence to clinical disease, therapeutic morbidity should not outpace risk of disease progression. Salvage radiation therapy following radical prostatectomy has widely variable long-term biochemical control rates (from 18 to 64% depending on the follow-up period). Early hormonal therapy delivered as castration or complete androgen blockade might delay clinical metastasis in patients with high-risk pathologic disease; however, the adverse effects and morbidity of long-term therapy must not be underestimated. Non-steroidal antiandrogens as monotherapy for early biochemical recurrence, particularly for younger men who wish to preserve their libido and sexual potency, have received considerable attention, but there are conflicting data on long-term outcomes. Because of their favorable adverse-effect profiles, non-traditional therapies that exert localized hormonal or cellular effects are receiving considerable attention for treatment of early, PSA-only recurrence. Data from animal models provide a rationale for the use of these therapies, but there is a lack of evidence to support prolongation of progression-free survival or cancer-specific survival. PMID- 16474761 TI - When and how to use informatics tools in caring for urologic patients. AB - Making predictions is an essential part of any medical decision. It is particularly crucial when considering treatment of clinically localized prostate cancer. Nomograms and prediction model software typically provide the most accurate predictions. Many nomograms have been developed, for all prostate cancer clinical states. Some of these are discussed in this review, as is their utility in facilitating decision making and informed consent. PMID- 16474762 TI - Technology insight: High-intensity focused ultrasound for urologic cancers. AB - The growing interest in high-intensity focused ultrasound (HIFU) technology is mainly due to its many potential applications as a minimally invasive therapy. It has been introduced to urologic oncology as a treatment for prostate and kidney cancers. While its application in the kidney is still at the clinical feasibility phase, HIFU technology is currently used in daily practice in Europe for the treatment of prostate cancer. Literature describing the results of HIFU for prostate cancer is mainly based on several series of patients from clinical development teams. The latest published results suggest that HIFU treatment is a valuable option for well-differentiated and moderately-differentiated tumors, as well as for local recurrence after external-beam radiation therapy. PMID- 16474764 TI - The learning curve as a measure of experience. PMID- 16474763 TI - A case of undiagnosed tethered cord syndrome aggravated by transurethral prostate resection. AB - BACKGROUND: A 68-year-old man presented with a history of significant urinary urge incontinence, pollakiuria, and weak bladder sensation. He also reported mild fecal incontinence and a hypotrophic and slightly weaker left leg. At 63 years of age he had presented to a urologist for treatment of irritative lower urinary tract symptoms and incontinence. A transurethral resection of the prostate had been performed. After the operation, the symptoms had persisted and the incontinence seriously worsened. INVESTIGATIONS: Clinical neurologic examination, videourodynamic examination, neurophysiologic examination, and MRI of the spinal cord. DIAGNOSIS: Neurogenic bladder dysfunction caused by adult tethered cord syndrome with myelon up to S2 level, spina bifida occulta, and lipoma infiltrating the conus medullaris. MANAGEMENT: Conservative anticholinergic treatment failed, and injection of botulinum-A toxin is planned. PMID- 16474765 TI - Overactive bladder: drug therapy versus nerve stimulation. PMID- 16474785 TI - The current status of laparoscopic retroperitoneal lymph node dissection for non seminomatous germ-cell tumors. AB - An appropriately performed retroperitoneal lymph node dissection (RPLND) is both a diagnostic and a therapeutic procedure in the management of non-seminomatous germ-cell tumors. In an attempt to minimize the morbidity associated with an open RPLND, several investigators have evaluated the role of laparoscopic RPLND. Laparoscopic RPLND is technically feasible in the hands of dedicated experts, and results in a reduced hospital stay, operative blood loss, and post-operative morbidity compared to open RPLND. Currently, however, none of the reports in the literature have adequately evaluated the therapeutic efficacy of laparoscopic RPLND, as the vast majority of patients are treated with adjuvant chemotherapy regardless of the risk of systemic relapse. Future prospective internal review board-approved trials need to be developed to evaluate the therapeutic efficacy of laparoscopic RPLND, with careful observation of patients with pathologic low volume nodal disease. PMID- 16474786 TI - The role of lymphadenectomy in prostate cancer. AB - It has been shown that an adequate lymphadenectomy for exact staging of prostate cancer consists of removal of all the tissue along the external iliac vein, in the obturator fossa and along the internal iliac artery. Morbidity associated with this procedure is low, if certain technical details are respected. This review discusses in detail the indications for lymphadenectomy and the extent of dissection, based on the localization of the positive nodes. The potential therapeutic impact of extended lymph node dissection in men with prostate cancer is also addressed. PMID- 16474787 TI - Therapy Insight: management of urology patients taking long-term warfarin anticoagulation therapy. AB - Chronic warfarin anticoagulation is commonly used to prevent thromboembolism in patients with atrial fibrillation or venous thromboembolism, and in the management of patients with mechanical heart valves. Interruption of long-term anticoagulation therapy for elective, planned urologic procedures in these patients creates a complex situation in which competing risks of thrombosis and hemorrhage must be managed; when anticoagulation is withheld patients are at risk of thrombosis, and when it is restarted they are at risk of hemorrhage. Patients at a high risk of thrombosis are typically given bridging therapy with heparin to reduce the amount of time without anticoagulation. Outcomes from bridging therapy are influenced by patient characteristics, including the risk of thromboembolism from underlying medical conditions and the risk of perioperative bleeding, and by characteristics of the procedures themselves. The safety and efficacy of different approaches are not well documented. Data regarding periprocedural anticoagulation management of urology patients is limited and principally describes outcomes related to transurethral prostate surgery. Results from these studies indicate that various strategies of anticoagulation interruption and bridging therapy result in low frequencies of thromboembolism, but variable rates of hemorrhage. Patients on anticoagulation therapy who are due to undergo invasive urologic procedures that have a low risk of developing thromboembolism can discontinue warfarin 4-5 days before the procedure, and resume it postprocedure. Bridging therapy to prevent thrombosis is necessary for patients at a higher risk of developing thromboembolism. Future research should develop strategies to maintain low rates of thrombosis but reduce the frequency of postoperative hemorrhage. PMID- 16474788 TI - Bilateral kidney stones with ureteropelvic junction obstruction. AB - BACKGROUND: A 56-year-old male with bilateral flank pain was admitted following diagnosis of bilateral kidney stone disease. He had previously undergone four separate pyelolithotomy sessions (two per kidney), the latest being 14 years prior. At presentation the patient had two stones in the right kidney and seven in the left kidney. The right renal pelvis was dilated, indicating obstruction of the ureteropelvic junction. INVESTIGATIONS: Plain radiography, ultrasonography, intravenous pyelography, retrograde ureteropyelography and spiral tomography. DIAGNOSIS: Bilateral kidney stones and obstruction of the right ureteropelvic junction. MANAGEMENT: Bilateral percutaneous nephrolithotomy and simultaneous right endopyelotomy; intraoperative antegrade nephrogram; preoperative and postoperative measurement of hemoglobin, blood urea nitrogen and creatinine; postoperative nephrostograms, plain X-ray, diuretic renogram and intravenous pyelography. PMID- 16474789 TI - So what about history? PMID- 16474810 TI - Therapy Insight: sexual and bladder dysfunction associated with diabetes mellitus. AB - Diabetes mellitus affects 3-6% of the population. Patients with diabetes experience chronic vascular complications, which lead to a wide range of medical problems. Genitourinary problems are included among these complications, related to both neuropathy and vasculopathy. The most important clinical features relating to genitourinary involvement in patients with diabetes include erectile dysfunction (ED) and retrograde ejaculation in men, and bladder dysfunction. There is a 36% prevalence of ED in men with diabetes, which is about three times higher than in the general population. With so many other factors contributing to the risk of developing ED, it is important that men with diabetes are advised not only regarding glycemic control, but also in relation to lifestyle factors that might contribute to the development of ED, such as smoking. Diabetes-associated bladder dysfunction, characterized by decreased bladder sensation, increased bladder capacity, and impaired detrusor contractility, is another important condition to consider when treating patients with diabetes. Accurate assessment is important in these patients in order to reduce the damage associated with neurogenic bladder dysfunction. Management goals for these patients include voiding strategies for relief of symptoms, prevention and treatment of infections, continence, and adequate bladder emptying. This review discusses the mechanisms behind urologic conditions that are often encountered when treating patients with diabetes, and summarizes the evaluation and management of these patients. PMID- 16474811 TI - Mechanisms of Disease: new insights into the cellular and molecular pathology of Peyronie's disease. AB - Peyronie's disease (PD) is characterized by fibrotic plaques in the penile tunica albuginea that cause curvature of the erect penis, and is often accompanied by pain and/or erectile dysfunction. This condition affects up to 9% of men. Treatment is mainly surgical, as pharmacologic therapy has limited efficacy. The pathophysiology of PD is poorly understood, but development of two rat models, extrapolation of what is known about the molecular pathology of other fibrotic conditions, and emphasis on the role of myofibroblasts and adult stem cells are helping to clarify etiology and identify new pharmacologic targets. Recent studies demonstrate a role for oxidative stress and cytokine release-primarily transforming-growth-factor beta1-in development of PD fibrotic plaques. There is evidence indicating that these profibrotic factors interact with antifibrotic defense mechanisms, such as decrease of myofibroblast accumulation, elimination of reactive oxygen species by inducible nitric oxide synthase and neutralization of transforming-growth-factor beta1 by decorin, such that some plaques are in dynamic turnover. Injury to the erect penis is thought to trigger PD by inducing extravasation of fibrin and subsequent synthesis of transforming-growth-factor beta1. Despite the lack of statistical support for a causal association between trauma and PD, it is possible that undetected microtrauma is involved. It is not known whether ossification of PD plaques is linked to fibrosis progression or is a manifestation of an alternative pathway. Both processes seem to be related to activation of fibroblast/myofibroblast differentiation in the tunica albuginea and to osteogenic commitment of stem cells in this tissue. PMID- 16474812 TI - Management of clinically localized prostate cancer by radical prostatectomy followed by watchful waiting. AB - BACKGROUND: A 65-year-old married man requested a PSA screening test and was found to have an elevated PSA level of 5.26 ng/ml. INVESTIGATIONS: Digital rectal examination, sextant biopsy, bone scan, and MRI to confirm diagnosis and stage the disease. Subsequent histopathologic examination of the excised prostate. DIAGNOSIS: Preoperative stage cT2b prostate cancer (Gleason score 7 [3 + 4]). Postoperative stage pT3b, N0, M0 prostate cancer (Gleason score 9 [4 + 5]), with extensive cancer within the left side of the prostate gland, involving several of the surgical margins and extending to the proximal portion of the left seminal vesicle. MANAGEMENT: Open radical retropubic prostatectomy, then watchful waiting with further treatment deferred until disease progression. Postoperative erectile dysfunction treated with sildenafil plus prostaglandin E(1) combination therapy. PMID- 16474813 TI - Locally advanced prostate cancer treated with radiotherapy and androgen deprivation. AB - BACKGROUND: A 60-year-old man, with a 2-year history of lower-urinary-tract symptoms (frequency and reduced flow) and perineal discomfort, presented with a serum PSA level of 42 ng/ml. INVESTIGATIONS: Digital rectal examination, transrectal ultrasound, prostate biopsy (8 cores), prostate and pelvic MRI, renal ultrasound and bone scan. DIAGNOSIS: cT3b, N0, M0 prostate cancer (Gleason score 7 [4 + 3]). MANAGEMENT: Hormonal down-staging with bicalutamide 150 mg/day for 3 months, then conformal radiotherapy (70 Gy over 7 weeks) with adjuvant bicalutamide 150 mg/day, to be continued until disease progression. Breast radiotherapy administered over a 5-day period at a dose of 15 Gy to reduce nonsteroidal antiandrogen-associated gynecomastia and breast pain. PMID- 16474814 TI - Artificial neural networks: has the time come for their use in prostate cancer patients? PMID- 16474815 TI - Who should be treating erectile dysfunction? PMID- 16474834 TI - Practical approaches to the diagnosis and management of male infertility. AB - Male infertility affects 10% of couples of reproductive age worldwide, and is treatable in many cases. In addition to well-established etiologies, genetic causes of male infertility are now diagnosed more commonly, as our knowledge of genomic medicine advances. Using principles of evidence-based medicine, this review outlines diagnostic and treatment algorithms that guide clinical management. In order of importance, randomized controlled clinical trials, basic scientific studies, meta-analyses, case-controlled cohort studies, best-practice policy recommendations and reviews from peer-reviewed literature were incorporated into algorithms that provide organized and timely guidelines to the current management of male infertility. The strength of the evidence for treatment recommendations is also classified when appropriate. PMID- 16474835 TI - Drug Insight: oral phosphodiesterase type 5 inhibitors for erectile dysfunction. AB - Erectile dysfunction (ED) is a common medical condition that affects the sexual life of millions of men. At present, first-line oral pharmacotherapy for most patients with ED is a phosphodiesterase type 5 (PDE-5) inhibitor, of which three are currently available worldwide. Sildenafil (Viagra, Pfizer) has a very satisfactory efficacy-safety profile in all patient categories. The first PDE-5 inhibitor to reach the market, it is now the most widely prescribed oral agent for ED. Tadalafil (Cialis, Lilly ICOS) and vardenafil (Levitra, Bayer/GlaxoSmithKline) were introduced to the European Union and the US in 2003 and 2004, respectively. These three PDE-5 inhibitors share many characteristics, but each has unique features. This review describes the chemical, pharmacologic and clinical features of sildenafil, vardenafil and tadalafil as oral first-line treatments for ED. First, we describe the physiology of penile erection and PDE-5 inhibitor pharmacology, including chemistry, PDE selectivity, pharmacokinetics, and possible drug interactions. We then summarize data on the efficacy and safety profiles of the three PDE-5 inhibitors for the treatment of ED in the general population, in patients with diabetes mellitus and in men that have undergone bilateral nerve-sparing retropubic radical prostatectomy. PMID- 16474836 TI - Management of von Hippel-Lindau-associated kidney cancer. AB - Von Hippel-Lindau disease (VHL) is an autosomal-dominant inherited condition that predisposes patients to develop renal cysts and tumors, most commonly in the second to fourth decades of life. Renal cysts and tumors have historically been a major cause of disease-related morbidity and mortality, so urologists are often called on to manage patients with VHL. Knowledge of the extrarenal manifestations of VHL (hemangioblastomas of the central nervous system and retina, endolymphatic sac tumors, pancreatic cysts, epididymal and broad-ligament cysts, and pheochromocytomas) and integration of nonurologic specialties into management teams for VHL patients will help to achieve successful outcomes. Screening for renal manifestations of VHL, by regular imaging of the abdomen, begins late in the second decade of life. Because renal tumors in VHL can be multifocal and bilateral, management can be complex. Radical nephrectomy removes all tissue at risk for forming renal tumors; however, this necessitates renal replacement therapy. In an effort to control cancer effectively while preserving native renal function and minimizing intervention, some researchers have proposed an observational strategy. Patients are screened until the largest tumor reaches 3 cm in diameter, at which time operative intervention is recommended. Nephron sparing surgery is undertaken, whenever technically feasible, with the goal of removing all tumors in that renal unit. The role of minimally invasive technologies is currently being evaluated in selected patients with VHL renal masses. Elucidation of molecular pathways associated with VHL renal tumors may facilitate development of effective medical treatments for these lesions in the future. PMID- 16474837 TI - Sleep-related painful erections. AB - BACKGROUND: A 45-year-old man presented with repeated awakenings at night caused by nocturnal painful erections, in sharp contrast with normal erections at other times, 2 years after surgical removal of a thoracic (Th6-7) ependymoma. INVESTIGATIONS: Physical examination, pharmaco-sensitized penile Doppler ultrasound, spinal and brain MRI, neurophysiological work-up (tibial and perineal somatosensory-evoked potential; hand, foot and perineal sympathetic skin response assessment), polysomnography with recording of nocturnal penile tumescence. DIAGNOSIS: Sleep-related painful erections, characterized by penile pain during nocturnal erection, typically during rapid eye movement sleep, in the presence of a residual thoracic spinal cord syndrome. MANAGEMENT: Amitriptyline, an antidepressant that suppresses rapid eye movement sleep, was ineffective. Treatments with other antidepressants, clozapine and beta-blockers were suggested, but the patient declined because of potential severe side effects. PMID- 16474839 TI - Glyceraldehyde 3-phosphate dehydrogenase is a SET-binding protein and regulates cyclin B-cdk1 activity. AB - We report here that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) interacts in vitro and in vivo with the protein SET. This interaction is performed through the acidic domain of SET located at the carboxy terminal region. On analysing the functional relevance of SET-GAPDH interaction, we observed that GAPDH reverses in a dose-dependent manner, the inhibition of cyclin B-cdk1 activity produced by SET. Similarly to SET, GAPDH associates with cyclin B, suggesting that the regulation of cyclin B-cdk1 activity might be mediated not only by the interaction of GAPDH with SET but also with cyclin B. To analyse the putative role of GAPDH on cell cycle progression, HCT116 cells were transfected with a GAPDH expression vector. Results indicate that overexpression of GAPDH does not affect the timing of DNA replication but induces an increase in the number of mitosis, an advancement of the peak of cyclin B-cdk1 activity and an acceleration of cell cycle progression. All these results suggest that GAPDH might be involved in cell cycle regulation by modulating cyclin B-cdk1 activity. PMID- 16474838 TI - The mutation in the ATP-binding region of JAK1, identified in human uterine leiomyosarcomas, results in defective interferon-gamma inducibility of TAP1 and LMP2. AB - The presentation of human leukocyte antigens (HLA) class I requires the coordinated expression of numerous components involved in antigen presentation. Tumor cells may alter the antigen presentation by HLA class I, allowing them to evade antitumor immunity. In many cases, the lack of antigen presentation can be attributed to the downregulation of genes needed for antigen processing, such as the transporters associated with antigen processing (TAP) 1, and the proteasomal component, low molecular weight proteins (LMP) 2. The TAP1 and LMP2 genes are transcribed from a shared bidirectional promoter containing an interferon (IFN) gamma-response factor element; thus, the IFN-gamma-signal strongly induces both TAP1 and LMP2 expression. Low molecular weight proteins2-deficient mice exhibited the development of uterine leiomyosarcomas. Here, the differential responsiveness to IFN-gamma of the SKN human uterine leiomyosarcomas cell line was investigated. We now identify the G871E mutation in the ATP-binding region of Janus kinases 1, suggesting that the loss of TAP1 and LMP2 induction is a defect in the earliest steps of the IFN-gamma-signal pathway, resulting in the inability of SKN cells to upregulate the antigen-processing pathway. Understanding the mechanisms by which these tumors circumvent cytokine signalling, thereby evading antitumor-specific immunity, would greatly aid the efficacy of immunotherapy for treating uterine leiomyosarcomas. PMID- 16474840 TI - Decreased intranuclear mobility of acute myeloid leukemia 1-containing fusion proteins is accompanied by reduced mobility and compartmentalization of core binding factor beta. AB - Acute myeloid leukemia 1 (AML1) gene on chromosome 21 is involved in several chromosomal translocations, including t(8;21) and t(16;21), that produce chimeric fusion proteins AML1-eight twenty-one (ETO) and AML-myeloid transforming gene chromosome 16 (MTG16), which contribute to leukemogenesis. The molecular basis for the leukemogenic effects of these fusion proteins is incompletely understood. Using gel-shift assay, we showed that AML1-ETO and AML1-MTG16 bound to a series of AML1 consensus DNA-binding sites with different affinities. Using fluorescence recovery after photobleaching (FRAP), we demonstrated that a fusion of AML1 with ETO or MTG16 exhibits reduced intranuclear mobility compared with wild-type AML1 or either fusion partner. The dimerization domain (nervy homology region 2) of ETO is responsible for the reduced mobility of AML1-ETO. Dual FRAP studies revealed that CBFbeta colocalized with AML1-ETO within the nucleus, resulting in reduced mobility of CBFbeta. Therefore, AML1 fusion proteins may interfere with normal AML1 function due to aberrant nuclear dynamics, which leads to spatial and temporal sequestration of CBFbeta and perhaps other coregulators critical for myeloid differentiation. PMID- 16474841 TI - Loss of Sef (similar expression to FGF) expression is associated with high grade and metastatic prostate cancer. AB - Fibroblast growth factors (FGF), and in particular FGF8, have been strongly implicated in prostate carcinogenesis. This study investigated the expression of Sef, a key inhibitory regulator of FGF signalling, in prostate cancer. In a panel of cell lines, hSef was detected in both androgen-dependent and independent cells but was significantly reduced in highly metastatic derivative clones. hSef expression was not influenced by androgenic stimulation. Forced downregulation of hSef by siRNA increased FGF8b induced cell migration (P=0.02) and invasion (P=0.007). Reduced hSef levels also enhanced FGF8b stimulated expression of MMP9 (P=0.005). mRNA in situ hybridization revealed hSef expression in 80% (8/10) of benign biopsies but in only 69% (23/33) of Gleason sum 4-7 and 35% (10/28) of Gleason sum 8-10 cancer biopsies (P=0.004). Quantitative PCR of microdissected glands confirmed this trend (P=0.001). hSef was expressed in 69% (27/39) of non metastatic tumours but in only 18% (2/11) of metastatic tumours (P=0.004, n=50). hSef expression was next correlated with earlier data on FGF8b expression in a subgroup of cancers. In this cohort, 86% (19/22) of high-grade cancers expressed FGF8 but only 31% (7/22) expressed hSef. Positive FGF8 expression but a loss of hSef was observed in 88% (7/8) of metastatic tumours. In contrast, metastasis was evident in only 10% (1/10) of tumours, which co-expressed both FGF8 and hSef (P<0.001). These results suggest evidence that hSef is downregulated in advanced prostate cancer and might facilitate an enhanced tumorigenic response to FGFs. Further research into the role of hSef in cancer cell signalling and the mechanism of its downregulation may contribute to more effective targeting of growth factors in prostate cancer. PMID- 16474842 TI - Oncogenic Vav1 induces Rac-dependent apoptosis via inhibition of Bcl-2 family proteins and collaborates with p53 deficiency to promote hematopoietic progenitor cell proliferation. AB - Vav1 is an hematopoietic-specific Rho guanine nucleotide exchange factor coupling tyrosine kinase receptors and Rac GTPases, and has been implicated in transformation of fibroblasts and pancreas. To determine the biologic effect and oncogenic potential of Vav1 in hematopoietic lineages, we stably express oncogenic mutant of Vav1 in primary bone marrow cells using retrovirus-mediated gene transfer. Contrary to the growth stimulatory effects observed in fibroblasts, oncogenic Vav1 inhibits hematopoietic stem cell/progenitor engraftment in vivo and progenitor cell expansion in vitro via inducing apoptosis. The oncogenic Vav1-induced apoptosis is associated with reduced expression of Bcl-2 and Bcl-xL proteins and effectively suppressed by transgenic overexpression of Bcl-2, suggesting Vav1-mediated signaling via Bcl-2 in apoptosis. Also, oncogenic Vav1 stimulates sustained activation of Rac GTPases and the biologic effects of oncogenic Vav1 are Rac-dependent. Further, when expressed in the p53-deficient cells, which express elevated Bcl-2 and Bcl-xL and are resistant to the apoptosis, oncogenic Vav1 enhances both proliferation and self-renewal of hematopoietic progenitor cells. These results demonstrate clear phenotypic differences between wild-type and p53(-/-) hematopoietic cells expressing oncogenic Vav1, and suggest oncogenic potential of Vav1-mediated pathways in primary hematopoietic cell when they collaborate with additional genetic hits that affect the p53 pathway. PMID- 16474843 TI - Recruitment of ATR to sites of ionising radiation-induced DNA damage requires ATM and components of the MRN protein complex. AB - ATM and ATR are two related kinases essential for signalling DNA damage. Although ATM is thought to be the principle kinase responsible for signalling ionising radiation (IR)-induced DNA damage, ATR also contributes to signalling this form of genotoxic stress. However, the molecular basis of differential ATM and ATR activation in response to IR remains unclear. Here, we report that ATR is recruited to sites of IR-induced DNA damage significantly later than activation of ATM. We show that ATR is recruited to IR-induced nuclear foci in G(1) and S phase of the cell cycle, supporting a role for ATR in detecting DNA damage outside of S phase. In addition, we report that recruitment of ATR to sites of IR induced DNA damage is concomitant with appearance of large tracts of single stranded DNA (ssDNA) and that this event is dependent on ATM and components of the Mre11/Rad50/Nbs1 (MRN) protein complex. PMID- 16474844 TI - Tumor suppressor p53 regulates heparanase gene expression. AB - Mammalian heparanase degrades heparan sulfate, the most prominent polysaccharide of the extracellular matrix. Causal involvement of heparanase in tumor progression is well documented. Little is known, however, about mechanisms that regulate heparanase gene expression. Mutational inactivation of tumor suppressor p53 is the most frequent genetic alteration in human tumors. p53 is a transcription factor that regulates a wide variety of cellular promoters. In this study, we demonstrate that wild-type (wt) p53 binds to heparanase promoter and inhibits its activity, whereas mutant p53 variants failed to exert an inhibitory effect. Moreover, p53-H175R mutant even activated heparanase promoter activity. Elimination or inhibition of p53 in several cell types resulted in a significant increase in heparanase gene expression and enzymatic activity. Trichostatin A abolished the inhibitory effect of wt p53, suggesting the involvement of histone deacetylation in negative regulation of the heparanase promoter. Altogether, our results indicate that the heparanase gene is regulated by p53 under normal conditions, while mutational inactivation of p53 during cancer development leads to induction of heparanase expression, providing a possible explanation for the frequent increase of heparanase levels observed in the course of tumorigenesis. PMID- 16474845 TI - Acquired chemoresistance in pancreatic carcinoma cells: induced secretion of IL 1beta and NO lead to inactivation of caspases. AB - Pancreatic cancer exhibits profound chemoresistance resulting either from pre existing (intrinsic) mechanisms, or from anticancer drug treatment itself (acquired chemoresistance). To identify molecular alterations leading to acquired chemoresistance, the chemosensitive pancreatic carcinoma cell line PT45-P1 was exposed to low-dose treatment with etoposide for 6 weeks. Afterwards, these cells (PT45-P1res) were much more resistant to high-dose treatment with anticancer drugs than parental cells. Among several differentially expressed genes in PT45 P1res cells, IL-1beta was most significantly upregulated, a finding in line with our previous observation that IL-1beta accounts for intrinsic chemoresistance of pancreatic carcinoma cells. Elevated IL-1beta expression in PT45-P1res cells was confirmed by real-time PCR and ELISA, and treatment with the IL-1 receptor antagonist restored drug-induced apoptosis. The increased IL-1beta secretion was accompanied by an elevated formation of nitric oxide (NO) and a NO-dependent inhibition of the etoposide-induced caspase-3/-7/-8/-9 activity. Caspase activation was restored either by the iNOS inhibitor 1400W, the reducing agent dithiothreitol or the IL-1 receptor antagonist, resulting in greater sensitivity towards anticancer drug treatment. Conversely, IL-1beta or the NO-donor SNAP decreased caspase activation and apoptosis in etoposide-treated PT45-P1 cells. These data confirm IL-1beta and NO as determinants of chemoresistance in pancreatic cancer, and indicate that the intrinsic and acquired chemoresistance rely to some extent on common molecular targets beneficial for improved therapeutical strategies. PMID- 16474847 TI - Regulation of iNOS by the p44/42 mitogen-activated protein kinase pathway in human melanoma. AB - Activating mutations of the genes for NRAS and BRAF, components of the p44/42 mitogen-activated protein kinase (MAPK) pathway, are common findings in melanoma. Recent evidence in several nonmelanoma cell systems supports the regulation of the inducible nitric oxide synthase (iNOS) gene by this pathway. On the basis of our data showing that melanoma iNOS expression predicts shortened patient survival, we formulated the hypothesis that activating mutations of NRAS or BRAF, which lead to constitutive activation of the p44/42 MAPK pathway, drive iNOS expression in human melanoma. In the present study, we have shown that inhibition of melanoma iNOS activity by S-methylisothiourea leads to decreased cell proliferation, confirming the importance of iNOS activity for melanoma cell growth. Regulation of melanoma iNOS expression by the p44/42 MAPK pathway was demonstrated by inhibition of the pathway by U0126, and by BRAF RNA interference. To explore this regulatory pathway in human tissue, 20 melanoma tumors were examined for NRAS and BRAF mutations, immunohistochemical evidence of ERK phosphorylation, and iNOS expression. A significant association was found among these three features. We conclude that in human melanoma, activating mutations of NRAS and BRAF drive constitutive iNOS expression and, implicitly, nitric oxide production, contributing to the poor survival of these patients. PMID- 16474846 TI - TAp63alpha indirectly regulates a cutaneous HPV promoter through complex formation with Jun family members. AB - The p63alpha isoforms of the p53 family have been demonstrated to play a crucial role in the development and differentiation of the skin. We show that expression of the TAp63alpha isoform leads to an upregulation of the cutaneous papillomavirus HPV 20 promoter, which is increased at least three-fold when c-Jun is co-expressed, in contrast to a minimal increase in activity in the presence of c-Jun alone. Co-expression of TAp63alpha with JunB or JunD, respectively, and in combination, leads to a reduction in the viral promoter activation measured by the expression of TAp63alpha alone. JunB and JunD also inhibits the additive effect exerted on the TAp63alpha activation by c-Jun. Co-immunoprecipitation assays demonstrate a complex formation of c-Jun, JunB and JunD with TAp63alpha through the SAM domain mediating protein-protein interactions, which is characteristic for p63alpha. Co-expression of p53 mutant R248W not only downregulates the differential modulation of the viral promoter by TAp63alpha alone and in the presence of the Jun family members, but leads to a reduction in the protein levels of the overexpressed c-Jun, JunB, JunD, as well as TAp63alpha. This model system provides insight into yet unknown pathways through which TAp63alpha and Jun may cooperate in the pathogenesis of HPV associated cutaneous lesions. PMID- 16474848 TI - ST7-mediated suppression of tumorigenicity of prostate cancer cells is characterized by remodeling of the extracellular matrix. AB - Multiple lines of evidence have provided compelling evidence for the existence of a tumor suppressor gene (TSG) on chromosome 7q31.1. ST7 may be the target of this genetic instability but its designation as a TSG is controversial. In this study, we show that, functionally, ST7 behaves as a tumor suppressor in human cancer. ST7 suppressed growth of PC-3 prostate cancer cells inoculated subcutaneously into severe combined immunodeficient mice, and increased the latency of tumor detection from 13 days in control tumors to 23 days. Re-expression of ST7 was also associated with suppression of colony formation under anchorage-independent conditions in MDA-MB-231 breast cancer cells and ST7 mRNA expression was downregulated in 44% of primary breast cancers. Expression profiling of PC-3 cells revealed that ST7 predominantly induces changes in genes involved in re modeling the extracellular matrix such as SPARC, IGFBP5 and several matrix metalloproteinases. These data indicate that ST7 may mediate tumor suppression through modification of the tumor microenvironment. PMID- 16474849 TI - Functional characterization of novel presenilin-2 variants identified in human breast cancers. AB - We identified in breast cancer cases two germline alterations, R62H and R71W, in presenilin-2 (PS-2), a gene involved in familial Alzheimer's disease (FAD). The role of these alleles in FAD is unclear, but neither allele affected Abeta(42)/Abeta(40) ratio. However, both R62H and R71W alterations compromised PS 2 function in Notch signaling in Caenorhabditis elegans and cell growth inhibition in mouse embryonic fibroblasts, and these effects were dependent on gene dosage. We found that both alterations enhanced the degradation of the PS-2 full-length protein, indicating that they may have a loss-of function effect. The effect of the R71W alteration was noticeably stronger, and we observed an almost threefold higher frequency of this allele in breast cancer cases versus controls, but this difference did not reach statistical significance. Nonetheless, these results collectively suggest that the novel PS-2 alleles described here, especially R71W, affect PS-2 function and may potentially confer a moderate risk of susceptibility to breast cancer. PMID- 16474850 TI - Complex regulation of human androgen receptor expression by Wnt signaling in prostate cancer cells. AB - beta-Catenin, a component of the Wnt signaling pathway, is a coactivator of human androgen receptor (hAR) transcriptional activity. Here, we show that Wnt signaling also influences androgen-mediated signaling through its ability to regulate hAR mRNA and protein in prostate cancer (PCa) cells. Three functional LEF-1/TCF binding sites lie within the promoter of the hAR gene as shown by CHIP assays that captured beta-catenin-bound chromatin from Wnt-activated LNCaP cells. Chimeric reporter vectors that use the hAR gene promoter to drive luciferase expression confirmed that these LEF-1/TCF binding elements are able to confer robust upregulation of luciferase expression when stimulated by Wnt-1 or by transfection with beta-catenin and that dominant-negative TCF or mutations within the dominant TCF-binding element abrogated the response. Semi-quantitative and real time RT-PCR assays confirmed that Wnt activation upregulates hAR mRNA in PCa cells. In contrast, hAR protein expression was strongly suppressed by Wnt activation. The reduction of hAR protein is consistent with evidence that Wnt signaling increased phosphorylation of Akt and its downstream target, MDM2 that promotes degradation of hAR protein through a proteasomal pathway. These data indicate that the hAR gene is a direct target of LEF-1/TCF transcriptional regulation in PCa cells but also show that the expression of the hAR protein is suppressed by a degradation pathway regulated by cross-talk of Wnt to Akt that is likely mediated by Wnt-directed degradation of the B regulatory subunit of protein phosphatase, PP2A. PMID- 16474851 TI - The tumour-suppressor protein ASPP1 is nuclear in human germ cells and can modulate ratios of CD44 exon V5 spliced isoforms in vivo. AB - The ASPP1 (Apoptosis Stimulating Protein of p53) protein is an important tumour suppressor. We have detected a novel protein interaction between the human ASPP1 (hASPP1) protein and the predominantly nuclear adaptor protein SAM68. In the human testis, full-length endogenous hASPP1 protein is located in the nucleus like SAM68, predominantly within meiotic and postmeiotic cells. Mouse ASPP1 (mASPP1) protein is mainly expressed in the brain and testis. The interaction with nuclear SAM68 is likely to be restricted to human germ cells, since endogenous mASPP1 protein is exclusively cytoplasmic. The C-terminal region of hASPP1 efficiently targeted a fused GFP molecule to the nucleus, whereas the N terminus of hASPP1 targeted GFP to the cytoplasm. In the context of the full length molecule this cytoplasmic targeting sequence is dominant in HEK293 and Saos-2 cells, since full-length hASPP1-GFP is almost exclusively cytoplasmic. Despite its predominantly cytoplasmic location, we show that ASPP1-GFP expression in HEK293 cells can regulate the ratio of alternative spliced isoforms derived from a pre-mRNA regulated downstream of cytoplasmic signalling pathways, and our data suggest that ASPP1 may operate in this case downstream or parallel to RAS signalling pathways. PMID- 16474853 TI - Preclinical and clinical development of the oral multikinase inhibitor sorafenib in cancer treatment. AB - Tumor survival, growth and metastasis depend on efficient tumor cell proliferation and tumor angiogenesis, and targeting both of these processes simultaneously could prove to be therapeutically relevant. The RAS/RAF signaling pathway is an important mediator of tumor cell proliferation, and angiogenesis and is often aberrantly activated in human tumors due to the presence of activated Ras or mutant B-Raf, or elevation of growth factor receptors. Sorafenib, which belongs chemically to a class that can be described as bis-aryl ureas, was selected for further pharmacologic characterization based on potent inhibition of Raf-1 and its favorable kinase selectivity profile. Further characterization showed that sorafenib suppresses both wild-type and V599E mutant B-Raf activity in vitro. In addition, sorafenib demonstrated significant activity against several receptor tyrosine kinases involved in neovascularization and tumor progression, including vascular-endothelial growth factor (VEGFR)-2, VEGFR 3, platelet-derived growth factor (PDGFR)-beta Flt-3, and c-KIT. Preclinically, sorafenib showed broad-spectrum antitumor activity in colon, breast and non-small cell lung cancer xenograft models. A total of four phase I studies using oral sorafenib as a single agent have been completed, and the compound showed a favorable safety profile with mild to moderate diarrhea being the most common treatment-related adverse event. The maximum tolerated dose was 400 mg b.i.d. continuous. Single-agent phase II trials reported so far demonstrated antitumor activity of sorafenib in patients with hepatocellular carcinoma, sarcoma and renal cell cancer (RCC). Based on phase II results in RCC patients, a placebo controlled phase III study was performed, which randomized a total of 905 patients, most of whom were treated previously. The partial response rate was 2% for sorafenib and 0% for placebo. Stable disease was observed in 78% and 55% of patients on sorafenib and placebo, respectively. Sorafenib significantly prolonged median progression-free survival (24 weeks) compared with placebo (12 weeks) in all subsets of patients evaluated. Approval of sorafenib by the U.S. Food and Drug Administration for this indication is pending. A first-line phase III study in RCC as well as phase III studies in hepatocellular carcinoma and metastatic melanoma have been initiated. PMID- 16474852 TI - Loss of SOCS3 in the liver promotes fibrosis by enhancing STAT3-mediated TGF beta1 production. AB - Recently, DNA methylation and reduced expression of the suppressor of the cytokine signaling-3 (SOCS3) gene in human hepatocellular carcinoma (HCC) patients have been reported. However, the roles of SOCS3 in HCC development in vivo have not been clarified. Using RT-PCR analysis and Western blotting, we confirmed that SOCS3 expression was reduced in HCC patients. However, reduced expression of SOCS3 occurred not only in HCC but also in nontumor regions, and this reduction was stronger as the fibrosis grade increased. Furthermore, SOCS3 levels were inversely correlated with signal transducers and activators of transcription-3 (STAT3) activation as well as transforming growth factor (TGF) beta1 levels in the non-HCC region. To define the molecular consequences of SOCS3 silencing/STAT3 hyperactivation and liver fibrosis, we examined liver-specific SOCS3-deficient mice. We demonstrated that SOCS3 deletion in the liver resulted in hyperactivation of STAT3 and promoted ConA- and chemical-induced liver fibrosis. The expression of TGF-beta1, a mediator of fibrosis, was enhanced by SOCS3 gene deletion, but suppressed by the overexpression of a dominant-negative STAT3 or SOCS3 both in vivo and in vitro. These data suggest that TGF-beta1 is a target gene of STAT3 and could be one of the mechanisms for enhanced fibrosis in SOCS3-deficient mice. Thus, our present study provides a novel role of SOCS3 and STAT3 in HCC development: in addition to the previously characterized oncogenic potentials, STAT3 enhances hepatic fibrosis through the upregulation of TGF-beta1 expression, and SOCS3 prevents this process. PMID- 16474854 TI - Rituximab. AB - Rituximab is a chimeric monoclonal antibody that targets the CD20 molecule on the B-cell surface. It is the first antibody of its kind to be licensed for the treatment of non-Hodgkin's lymphoma. Rituximab was found to be effective, well tolerated and has a good safety profile, though its precise cellular effects are still not well understood. Rituximab has been shown to be of therapeutic benefit in various autoimmune diseases in which B lymphocytes play a role. This article summarizes the current literature regarding the use of rituximab in lymphoma, rheumatoid arthritis, systemic lupus erythematosus and other selected autoimmune diseases. PMID- 16474855 TI - Pharmacogenetics of therapies in rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is an inflammatory, aggressive arthritis causing irreversible joint destruction and damage when left untreated. Disease-modifying antirheumatic drugs (DMARDs) are the mainstay of treatment for RA and ameliorate not only the clinical signs and symptoms but also joint damage associated with the disease. In recent years, biological therapies have been introduced for the treatment of RA, and the effectiveness of these agents in slowing the clinical and radiographic progression in RA has been established beyond question. However, there is significant variability in the response of patients with RA to these therapies. Moreover, the biological therapies are expensive, totaling several thousand dollars in yearly patient costs. Pharmacogenomics, the study of genetic variations in drug-metabolizing enzymes and their translation to differential responses to drugs, is a nascent but rapidly evolving field. The application of pharmacogenomics to therapies used in RA, particularly the new expensive biological agents, holds great promise for tailoring therapy with these agents based on a patient's genetics. Published literature on the pharmacogenetics of commonly used DMARDs and the emerging body of literature on the pharmacogenetics of the new biological therapies in RA are the focus of this review. As evident from the contents of this review, pharmacogenomics is an exciting field which is progressing productively and rapidly. Pharmacogenomic approaches offer powerful tools to optimize drug therapy in individual patients. PMID- 16474856 TI - Cerebrovascular disease and dementia. AB - Cerebrovascular disease and dementia are extremely prevalent and disabling disorders affecting older people. Results of previous pathological investigations and later epidemiological studies have raised the possibility that the two disorders may be causally related. The study of such causal associations may provide insights that could lead to the development of strategies intended to prevent or treat dementia more effectively. Cerebrovascular disease has many manifestations, some of which are strong causal factors in the development of a future dementia. However, uncertainty and controversy exist regarding the presence and nature of the causal contribution of others. Potential therapeutic strategies for dementia are hindered by the lack of understanding of such relationships and the consequent difficulty in identifying a clear phenotype of dementia occurring predominantly due to cerebrovascular disease. The field is ripe for further examination of the associations between vascular factors and dementia, and the mechanisms underlying such associations. The interface between basic and clinical science has much to offer in clarifying the relationships between aging, vascular factors and cognitive decline in older people. In this review, we will attempt to synthesize data available from epidemiological, clinical and basic science research in the field of dementia related to cerebrovascular disease, highlighting potential avenues for further research. PMID- 16474857 TI - [Waiting lists: an unsolved social phenomenon]. PMID- 16474858 TI - [Why is surgical innovation so badly needed]. PMID- 16474859 TI - [Ross procedure: mid term results]. AB - The Ross procedure has been used in children and young adults for aortic valve replacement and the correction of complex obstruction syndromes of the left ventricular outflow tract. We report the mid-term results of the Ross procedure in a single institution and performed by the same surgical team. POPULATION: Between March 1999 and December 2005, 18 patients were operated on using the Ross procedure. The mean age at the time of surgery was 12 years, being 12 patients male (67%). The primary indication for surgery was isolated aortic valve disease, being the predominant abnormality in 58% of cases aortic regurgitation and in 42% left ventricular outflow tract obstruction. Associated lesions included sub aortic membrane in 3 patients (16%), small VSD in 2 patients (11%), bicuspid aortic valve in 4 patients (22%) and severe left ventricular dysfunction and mitral valve regurgitation in 1 patient (6%). Ten of the 18 patients (56%) had been submitted to previous surgical procedures or percutaneous interventions. RESULTS: Early post-operative mortality was not seen, but two patients (11%), had late deaths, one due to endocarditis, a year after the Ross procedure, and the other due to dilated cardiomiopathy and mitral regurgitation. The shortest time of follow-up is 6 months and the longest 72 months (median 38 months). Of the 16 survivors, 14 patients are in class I of the NYHA and 2 in class II, without significant residual lesions or need for re-intervention. The 12 patients with more than a year of follow up revealed normal coronary perfusion in all patients and no segmental wall motion abnormalities. Nevertheless, two of the 12 patients developed residual dynamic obstruction of LVOT and in three patients aortic regurgitation of a mild to moderate degree was evident. Significant gradients were not verified in the RVOT. CONCLUSIONS: The Ross procedure, despite its complexity, can be undertaken with excellent immediate results. Aspects such as the dilation of the neo aortic root and homograft evolution can not be considered in a study of this nature, seeing that the mean follow up time does not exceed 5 years. PMID- 16474860 TI - [Ventricular assist in children]. AB - We describe the use of the Berlin Heart biventricular mechanical assistance device, as a bridge to transplant in a two-years old child suffering from end stage dilated cardiomiopathy. Ventricular support lasted for 3,5 months and led to successful transplantation. We describe the clinical case, the protocols used as well as present indications, techniques and problems related to the use of mechanical heart support in children. PMID- 16474861 TI - [Surgical diagnosis of mediastinal pathology: eleven-years experience]. AB - Between January 1993 and December 2004, 68 patients with mediastinal pathology were submitted to diagnosis surgery. All the pathological process were located in the anterior mediastinum, and most of them were malignant (64.1%). The most common pathology was limphoma (26.5%), sarcoidosis (23.5%) and thymomas (23.5%). Seven cases consisted in ganglionar tuberculosis (10.3%), six cases were ganglionar metastases of lung adenocarcinoma (8.8%), and three cases were thymic carcinoma (4.4%). One patient had no definite diagnosis (1.5%). In this experience high diagnosis accuracy (98.5%), and low morbidity were demonstrated. Although the present video assisted thoracic surgery progress and development, mediastinoscopy still remains as a reliable, safe and efficient surgical approach for the diagnosis of mediastinal pathology. PMID- 16474862 TI - [Revascularization surgery in infected fields: on the search for the ideal solution]. AB - The surgical management of aorto-iliac prosthetic infections and infectious aneurysms still remains nowadays as a serious and controversial issue, coursing with high morbi-mortality rates and having not reached completely satisfactory or consensual therapeutic solutions. Among the diverse surgical alternatives, the author enhances the extra-anatomic revascularization and the "in situ" procedures, which are different according to the material employed, including the prosthetic grafts, the arterial and venous autografts and the cryopreserved arterial allografts, describing its indications, advantages and disadvantages and biological behavior of these diverse materials commonly employed in the revascularization surgery in infected fields. PMID- 16474863 TI - [Long-term evaluation of chronic diabetic foot ulcers, non-healed after hyperbaric oxygen therapy]. AB - AIM OF THE STUDY: To assess the effect of systemic hyperbaric oxygen therapy in chronic diabetic lower limb lesions, non-healed after this adjunctive treatment, a retrospective controlled study was undertaken. RESEARCH DESIGN AND METHODS: From 1990 to 2003, 96 patients demonstrating chronic Wagner grades II-IV lower limb ulcers, with no improvement over a 6 month average period (range 1-48 m) of full standard treatment, were studied. 55 patients received HBOT (study group); Hyperbaric oxygen (HBO) was applied in a multiplace hyperbaric oxygen chamber, at 2.5 absolute atmospheres, during 90 minutes, once a day, 5 days a week; The average number of HBO sessions per patient was 54 (range 20-151). 41 patients refused HBOT or left the Hyperbaric Medical Center after first consultation. So, they did not receive HBOT (control group). HBO and control patients did not differ in their baseline characteristics (i.e., age, sex, type and duration of diabetes, type and duration of lower limb ulcers). RESULTS: 78% of the control group were followed over a mean period of 55 months; 61.8% of the HBO group patients were evaluated over a mean period of 45 months; The patients were assessed for wound healing and need for amputation. The mean healing rate of chronic ulcers was significantly higher in the HBO group; The need for amputation was significantly less in the HBO group; The mean rate of major amputation was less (about one half) in the HBO group, but it was statistically no significant; The need for minor amputation was less (about two thirds) in the HBO group, but it was statistically no significant. CONCLUSIONS: This retrospective study provides evidence that HBOT increased significantly (13 times more) the mean healing rate of chronic lower limb ulcers in diabetic patients, over a mean follow-up period of 45 months. It also provides evidence that this adjunctive therapy decreased significantly (2 times less) the need for amputation in these patients, over the same period of time. The need for major amputation was also less (about two times) in the HBO group, but statistically no significant; The mean time required for amputation was higher in the HBO group, but statistically no significant. The long-standing beneficial effects of HBOT may be explained by the sustained improvement of fibroblast collagen production and of the microvascular supply inside the leg ulcers, enhanced by this adjunctive therapy. PMID- 16474864 TI - [Traumatic vasculo-orthopedic combined lesions: 18-years retrospective evaluation of epidemiology and risks factors for amputation]. AB - This study was undertaken to assess retrospectively the epidemiology and risk factors for amputation of combined vasculo-orthopaedic traumatic lesions, during a 18-year period, from March 1987 to May 2005, comprising the review of the clinical charts of 149 patients. The series includes a predominance of male patients (84%) with an average age of 34 years. Eighty-five per cent of the lesions resulted from traffic accidents (49% velocipedes, 39% automobiles, 12% trampling), 10% were consequence of falls and 5% resulted from agriculture activities. Orthopaedic lesions include 83 fractures of upper limbs (22 open), 123 of lower limbs (67 open), 21 scapulo-thoracic dissociations and 27 knee dislocations. Vascular lesions include 8 subclavian, 20 axillary, 25 braquial, 10 radial, 6 ulnar and 2 diverse, in the upper limbs; and in lower limbs 4 iliac, 27 femoral, 52 popliteal, 5 tibio-peroneal trunk, 5 anterior tibial, 12 posterior tibial and 12 peroneal. Overall mortality was 3.3% and the amputation rate was 17%. Knee dislocations were responsible for one third of amputations, followed by fractures of bone legs. The association of femur to bone legs fractures from one side and of the knee to bone legs fractures from the other, coursed with high levels of amputation, together with lacerations and crushing of the limbs. The prompt diagnosis of the lesions and the immediate, hierarchic and multidisciplinary approach were considered also as relevant prognostic factors in the management of these most demanding conditions. PMID- 16474866 TI - [Traumatic rupture of the innominate artery]. PMID- 16474865 TI - [Traumatic pseudoaneurysm of the abdominal aorta. Case report]. AB - Abdominal aortic pseudoaneurysms are rare in clinical practice, mainly when they are related to a blunt abdominal trauma. Clinical presentation is often vague, but they can cause symptoms related to the compression of adjacent organs or structures. The authors report the clinical case of a man, aged 47 years old, who developed a pseudoaneurysm of the terminal aorta, following an abdominal blunt trauma, consequence of a traffic accident, who underwent successful surgical treatment. PMID- 16474867 TI - Static and dynamic aspects of DNA charge transfer: a theoretical perspective. AB - In this work, we approach the impact of dynamic and static disorder on DNA charge transfer from a theoretical and numerical perspective. Disordered or defect geometries are either realized via molecular dynamics simulations using a classical force field or by experimentally determined DNA bulge structures. We apply a chemically specific, atomically resolved extended Su-Schrieffer-Heeger model to compute the energy parameters relevant to DNA charge transfer. For both models studied here, the effective donor-acceptor couplings--and hence the charge transfer rates--significantly depend upon the geometry. Dynamic disorder leads to a correlation time in this quantity of the order of 30 fs, and the transfer rates universally exhibit a broad, yet well-defined, exponential distribution. For DNA bulges, the angle characterizing the defect controls the charge transfer efficiency. The results are discussed and extensively compared to experimental findings and other calculations. PMID- 16474868 TI - A highly sensitive method for time-resolved detection of O(1D) applied to precise determination of absolute O(1D) reaction rate constants and O(3P) yields. AB - We demonstrate detection, in the gas-phase, of O(1D2) at concentrations down to 10(7) cm(-3) and develop this new method for time-resolved kinetic studies allowing both the total removal rate of O(1D2), of up to 1.5 x 10(6) s(-1), and the fraction quenched to O(3P(J)) by species X, k(q)/k(X), to be determined precisely from a single time profile: at 295 K we find, k(O(1D2) + N2O) = (1.43 +/- 0.08) x 10(-10) cm3 s(-1) with k(q)/k(N2O) = 0.056 +/- 0.009; k(O(1D2) + C2H2) = (3.1 +/- 0.2) x 10(-10) cm3 s(-1) with k(q)/k(C2H2) = 0.020 +/- 0.010; k(q)/k(H2O) < 0.003 for O(1D2) + H2O. PMID- 16474869 TI - Electrochemical multi-tagging of cysteinyl peptides during microspray mass spectrometry: numerical simulation of consecutive reactions in a microchannel. AB - On-line electrogeneration of mass tags in a microspray emitter is used to quantify the number of cysteine groups in a given peptide. A finite-element simulation of the multi-step process yields the relative distribution and concentration of tags, untagged and tagged species in the microchannel before the spray event. The work focuses on the tagging of cysteine moieties in peptides or proteins by electrogenerated quinone mass probes. The main chemical parameters determining the kinetics of the labelling are assessed and discussed considering the microfluidic aspects of the process. The control of the tagging extent allows the simultaneous MS analysis of both the unmodified and modified peptide(s). The number of cysteine groups corresponds to the number of characteristic mass shifts observed from the unmodified peptide. The present theoretical work establishes the range of optimum conditions for the determination of the number of cysteine groups in peptides containing up to five cysteine groups. PMID- 16474870 TI - Evaluation of the field-adapted ADMA approach: absolute and relative energies of crambin and derivatives. AB - A large number of conformations and chemically modified variants of the protein crambin were used to extensively test the field-adapted adjustable density matrix assembler (FA-ADMA) method developed for ab initio quality quantum chemistry computations of proteins and other macromolecules, introduced in an earlier publication. In this method, the fuzzy density matrix fragmentation scheme of the original adjustable density matrix assembler (ADMA) method has been made more efficient by combining it with an approach of using point charges to approximate the effects of additional, distant parts of a given macromolecule in the quantum chemical calculation of each fragment. In this way, smaller parent molecules can be used for fragment generation, while achieving accuracy that can be obtained only with large parent molecules in the original ADMA method. Whereas in both methods the error relative to the Hartree-Fock result can be reduced below any threshold by choosing large enough parent molecules, this can be done more efficiently with the new method. In order to obtain reliable test results for the accuracy obtainable by the new method when compared to conventional Hartree-Fock calculations, we performed a large number of energy calculations for the protein crambin using various conformations available in the Protein Data Bank, various protonation states, and side chain mutations. Additionally, in order to test the performance of the method for protein-solvent interaction studies, the energy changes due to the formation of complexes with ethanol and single and multiple water molecules were investigated. PMID- 16474871 TI - Photoinduced intramolecular charge transfer to meta position of benzene ring in 6 aminophthalides. AB - Substitution of non-fluorescent phthalide (Pd) with amino group at meta (6) position in relation to the electron-accepting part of the lactone ring completely changes Pd photophysics: a new long-wavelength absorption band arises and the molecule becomes highly fluorescent. The experimental data and the analysis of vertical electronic transitions with TDDFT method indicate that the first absorption band in 6-aminophthalides (6-APds) comprises a single CT transition to the S1 state. Almost equal absorption and emission transition dipole moments indicate that S0 <--> S1 transition in all 6-APds is not affected by any mixing with other electronic states, the excited-state vibrational relaxation is not accompanied by significant conformational changes and the Stokes shifts reflect mainly solvation energetics of these molecules. Excited state dipole moments obtained from solvatochromic plots and from CASSCF calculations confirm large charge displacement from amino group towards the meta position of the benzene ring upon excitation of 6-APds to S1 state. Long fluorescence lifetimes and high fluorescence quantum yields demonstrate efficient and stable excited state charge separation in 6-APds. Taken together with sensitivity of 6-APds to polarity and proticity of the environment these properties make them good candidates for fluorescent probes of long-time scale molecular dynamics. PMID- 16474872 TI - Soft or hard ionization of molecules in helium nanodroplets? An electron impact investigation of alcohols and ethers. AB - Electron impact (70 eV) mass spectra of a series of C1-C6 alcohols encased in large superfluid liquid helium nanodroplets (approximately 60,000 helium atoms) have been recorded. The presence of helium alters the fragmentation patterns when compared with the gas phase, with some ion product channels being more strongly affected than others, most notably cleavage of the C(alpha)-H bond in the parent ion to form the corresponding oxonium ion. Parent ion intensities are also enhanced by the helium, but only for the two cyclic alcohols studied, cyclopentanol and cyclohexanol, is this effect large enough to transform the parent ion from a minor product (in the gas phase) into the most abundant ion in the helium droplet experiments. To demonstrate that these findings are not unique to alcohols, we have also investigated several ethers. The results obtained for both alcohols and ethers are difficult to explain solely by rapid cooling of the excited parent ions by the surrounding superfluid helium, although this undoubtedly takes place. A second factor also seems to be involved, a cage effect which favors hydrogen atom loss over other fragmentation channels. The set of molecules explored in this work suggest that electron impact ionization of doped helium nanodroplets does not provide a sufficiently large softening effect to be useful in analytical mass spectrometry. PMID- 16474873 TI - A matrix isolation study of the C(s) symmetric OCNO(X 2A'') radical. AB - Here we report the first experimental detection of the C(s) symmetric nitroformyl radical, OCNO(X 2A'') in a nitrogen-carbon dioxide matrix at 10 K using a Fourier transform infrared spectrometer (FTIR). The nu1 vibrational frequency was observed at 2113 cm(-1). This assignment was confirmed by follow-up experiments using isotopically labeled reactant molecules (15N, 18O, 13C). To synthesize this radical, we irradiated solid nitrogen-carbon dioxide ice mixtures with energetic electrons at 10 K. Suprathermal nitrogen atoms in their electronic ground and/or first electronically excited state were generated via the radiation induced degradation of molecular nitrogen; these atoms could then react with carbon dioxide to eventually yield the nitroformyl radical. We also investigated the kinetics of the formation of the nitroformyl radical and support the arguments with computations on the doublet and quartet OCNO potential energy surfaces (PESs). PMID- 16474874 TI - Optical resolution of oriented enantiomers via photodissociation: quantum model simulations for H2POSD. AB - We demonstrate quantum mechanically how to resolve enantiomers from an oriented racemic mixture taking advantage of photodissociation. Our approach employs a femtosecond ultraviolet (UV) laser pulse with specific linear polarization achieving selective photodissociation of one enantiomer from a mixture of L and R enantiomers. As a result, the selected enantiomer is destroyed in the electronically excited state while the opposite enantiomer is left intact in the ground state. As an example we use H2POSD which presents axial chirality. A UV pulse excites the lowest singlet excited state which has nsigma* character and is, therefore, strongly repulsive along the P-S bond. The model simulations are performed using wavepackets which propagate on two dimensional potential energy surfaces, calculated along the chirality and dissociation reaction coordinates using the CASSCF level of theory. PMID- 16474875 TI - Kinetic studies of the ClO + ClO association reaction as a function of temperature and pressure. AB - The kinetics of the association reaction of ClO radicals: ClO + ClO + M --> Cl2O2+ M (1), have been investigated as a function of temperature T between 206.0 298.0 K and pressure p between 25-760 Torr using flash photolysis with time resolved UV absorption spectroscopy. ClO radicals were generated following the photolysis of Br2/Cl2O mixtures in nitrogen diluent gas. Charge coupled device (CCD) detection of time resolved absorptions was used to monitor ClO radicals over a broad wavelength window covering the ClO (A 2Pi<-- X 2Pi) vibronic absorption bands. The high pass filtered ClO absorption cross sections were calibrated as a function of temperature between T = 206.0-320 K, and exhibit a negative temperature dependence. The ClO association kinetics were found to be more rapid than those reported in previous studies, with limiting low and high pressure rate coefficients, in nitrogen bath gas, k0 = (2.78 +/- 0.82) x 10(-32) x (T/300)(-3.99 +/- 0.94) molecule(-2) cm6 s(-1) and k(infinity) = (3.37 +/- 1.67) x 10(-12) x (T/300)(-1.49 +/- 1.81) molecule(-1) cm3 s(-1), respectively, (obtained with the broadening factor F(c) fixed at 0.6). Errors are 2sigma. The pressure dependent ClO association rate coefficients (falloff curves) exhibited some discrepancies at low pressures, with higher than expected rate coefficients on the basis of extrapolation from high pressures (p > 100 Torr). Reanalysis of data excluding kinetic data recorded below p = 100 Torr gave k0 = (2.79 +/- 0.85) x 10(-32) x (T/300)(-3.78 +/- 0.98) molecule(-2) cm6 s(-1) and k(infinity) = (3.44 +/- 1.83)x 10(-12) x (T/300)(-1.73 +/- 1.91) molecule(-1) cm3 s(-1). Potential sources of the low pressure discrepancies are discussed. The expression for k(0) in air bath gas is k0 = (2.62 +/- 0.80) x 10(-32) x (T/300)(-3.78 +/- 0.98) molecule(-2) cm6 s(-1). These results support upward revision of the ClO association rate coefficient recommended for use in stratospheric models, and the stratospheric implications of the results reported here are briefly discussed. PMID- 16474876 TI - Conformational and photophysical studies on porphyrin-containing donor-bridge acceptor compounds. Charge separation in micellar nanoreactors. AB - Photophysical studies with semi-rigid, 1, and flexible, 2, donor-bridge-acceptor (D-b-A2+) molecules with D a porphyrin and A2+ a methyl viologen moiety, were performed in neat polar solvents as well as included in surfactant (DTAB) aqueous and in reverse AOT/n-alkane micelles. The micelles acted as nanoreactors for the photoinduced electron transfer reaction upon laser excitation. In spite of the longer lifetime of the charge separated (CS) state in the semi-rigid tetrad 1(ca. 200 ns vs. ca. 100 ns for the flexible dyad 2), the CS formation quantum yield, for example in acetonitrile, was lower for the former (phi(CS) = 0.13) than for the latter (0.58). Comparison of the time-resolved fluorescence data in neat solvent and in the micelles yielded the phi(CS) values in the dilute micellar solutions. Application of laser-induced optoacoustic spectroscopy at various temperatures to 1 dissolved in a polar organic solvent (benzonitrile, BZN) included in aqueous DTAB nanoreactors afforded structural volume changes for the production in hundreds of ps of the CS state upon excitation of a polar molecule. The contraction during CS formation upon excitation of the collapsed conformer in BZN is attributed to the entering of solvent into the open molecular cavity. The opening upon formation of the CS state due to photoinduced electron transfer in the 1 collapsed conformation arises from the repulsion of the two positively charged ends in this state, as previously calculated. Inclusion of 1 in reverse AOT micelles in various n-alkanes also led to a contraction upon excitation, but the data had much more error due to the limited range of variability of the ratio of thermoelastic parameters. The data obtained with the more flexible "supermolecule" 2 showed the predicted large conformation flexibility of these molecules. PMID- 16474878 TI - Criticality of a liquid-vapor interface from an inhomogeneous integral equation theory. AB - A microscopic theory is developed to study the liquid-vapor interfacial properties of simple fluids with ab initio treatment of the inhomogeneous two body correlation functions, without any interpolation. It consists of the inhomogeneous Ornstein-Zernike equation coupled with the Duh-Henderson-Verlet closure and the Lovett-Mou-Buff-Wertheim equation. For the liquid-vapor interface of the Lennard-Jones fluid, we obtained the density profile and the surface tension, as well as their critical behaviour. In particular, we identified non classical critical exponents. The theory accurately predicts the phase diagram and the interfacial properties in a very good agreement with simulations. We also showed that the method leads to true capillary-wave asymptotics in the macroscopic limit. PMID- 16474877 TI - Adsorption of DNA and electric fields decrease the rigidity of lipid vesicle membranes. AB - The adsorption of calf-thymus DNA-fragments of 300 +/- 50 base pairs (bp) to the outer membrane monolayer of unilamellar lipid vesicles in the presence of Ca2+ ions has been quantified by the standard method of chemical relaxation spectrometry using polarized light. The vesicles of radius a = 150 +/- 45 nm are prepared from bovine brain extract type III containing 80-85% phosphatidylserine (PS) and palmitoyl-oleoyl-phosphatidylcholine (POPC) in the molar ratio PS : 2POPC; total lipid concentration [L(t)] = 1 mM in 1 mM HEPES buffer, pH 7.4 at T = 293 K (20 degrees C). The turbidity relaxations of vesicle suspensions, at the wavelength lambda = 365 nm at two characteristic electric field strengths are identified as electroelongation of the whole vesicle coupled to smoothing of thermal membrane undulations and membrane stretching, and at higher fields, to membrane electroporation (MEP). The elongation kinetics indicates that the DNA adsorption renders the membrane more flexible and prone to membrane electroporation (MEP). Remarkably, it is found that the Ca-mediated adsorption of DNA (D) decreases both, bending rigidity kappa and stretching modulus K, along an unique Langmuir adsorption isotherm for the fraction of bound DNA at the given Ca concentration [Ca(t)] = 0.25 mM. The characteristic chemo-mechanical parameter of the isotherm is the apparent dissociation equilibrium constant K(D,Ca) = 100 +/- 10 microM (bp) of the ternary complex DCaB of DNA base pairs (bp) and Ca binding to sites B on the outer vesicle surface. Whereas both kappa and K decrease in the presence of high electric fields (E), the key parameter K(D,Ca) is independent of E in the range 0 < or = E/(kV cm(-1)) < or = 40. PMID- 16474879 TI - Role of water molecules in the KcsA protein channel by molecular dynamics calculations. AB - Molecular dynamics simulations supported by electrostatic calculations have been conducted on the KcsA channel to determine the role of water molecules in the pore. Starting from the X-ray structure of the KcsA channel in its closed state at 2.0 angstroms resolution, the opening of the pore towards a conformation built on the basis of EPR results is studied. We show that water molecules act as a structural element for the K+ ions inside the filter and the hydrophobic cavity of the channel. In the filter, water tends to enhance the depth of the wells occupied by the K+ ions, while in the cavity there is a strong correlation between the water molecules and the cavity ion. As a consequence, the protein remains very stable in the presence of three K+ ions in the selectivity filter and one in the cavity. The analysis of the dynamics of water molecules in the cavity reveals preferred orientations of the dipoles along the pore axis, and a correlated behavior between this dipole orientation and the displacement of the K+ ion during the gating process. PMID- 16474880 TI - Effect of vertical temperature variation on the oscillatory wetting instability in a fluid Ga-Pb alloy. AB - Oscillatory wetting instabilities driven by capillary-gravitation forces have been explored very recently in the binary fluid Ga-Pb alloy [A. Turchanin, R. Tsekov and W. Freyland, J. Chem. Phys., 2004, 120, 11 171]. This system is characterized by a complete wetting transition at liquid-liquid coexistence. Due to its metallic nature the bulk and interfacial instabilities are strongly coupled via variation of the respective emissivities. In our previous work we have investigated these phenomena at different cooling cycles and at constant temperature inside the miscibility gap. In this study we present for the first time the observations of the oscillatory wetting instabilities also in heating cycles. The interfacial properties of a Ga0.95Pb0.05 alloy at conditions inside the miscibility gap have been investigated by following the second harmonic generation (SHG) intensity changes. Corresponding model calculations of the Pb rich wetting film instabilities have been performed taking into account the effect of a temperature variation vertical to the bulk sample. The influence of this temperature variation on the occurrence of the oscillations is discussed. PMID- 16474881 TI - The interaction of dolomite surfaces with metal impurities: a computer simulation study. AB - This study investigates the behaviour of selected, morphologically important surfaces of dolomite (CaMg(CO3)2), using computational modelling techniques. Interatomic potential methods have been used to examine impurity substitution at cationic sites in these surfaces. Environmentally prevalent cations were studied to this end, namely Ni2+, Co2+, Zn2+, Fe2+, Mn2+ and Cd2+, all of which are also found as end-member carbonate minerals. Solid-solution substitution was investigated and showed that Cd and Mn will substitute from their end-member carbonate phase at either dolomite cation site. Mn is found to preferentially substitute at Mg sites, in agreement with experimental findings. For Ni2+, Co2+ and Zn2+, the magnitude of substitution energies is approximately equal for all surfaces, with the exception of the (1014) surface. However, for the larger cations, a far greater disparity in substitution energies is observed. At a stepped surface, analogous substitutions were performed and it was found that substitution energies for all impurity cations were reduced, indicating that uptake is more viable during growth. The predominant surface, the (1014), was solvated with a monolayer of water in order to investigate the influence of hydration on substitution energetics. The addition of water changes the relative preference for substitution of the different cations. Under aqueous conditions, the substitution energy is determined by three competing factors, the relative importance of which cannot be predicted without this type of computational investigation. PMID- 16474882 TI - Dye-sensitized TiO2 nanotube solar cells: fabrication and electronic characterization. AB - TiO2 nanotubes (TNTs) with large aspect ratio and large specific surface area were prepared from P25 (Nippon Aerosil) and applied to dye-sensitized titanium dioxide solar cells (DSSCs). Optimization of fabrication conditions, i.e., pH of the starting paste, sintering temperature for the TiO2 electrodes, electrolyte compositions of DSSCs gave the high conversion efficiency with improved open circuit voltage (V(oc)) and fill factor (FF) when compared to DSSCs made of P25. The evaluation of dye adsorption and the photo-injected electron transport such as electron diffusion coefficient (D) and electron lifetime (tau) in TNTs electrodes revealed that the higher efficiency resulted from increase of electron density with keeping much longer tau in TNTs electrodes than in P25 electrodes. PMID- 16474883 TI - Application of neutral amidines and guanidines in coordination chemistry. AB - The aim of this perspective is to highlight the continued development and application of amidines and guanidines as neutral, N-based donor ligands in coordination chemistry. From a sporadic interest dating back to the 1980s, work on these two closely related classes of compounds has steadily grown, with examples of evermore sophisticated substitution about the common ligand framework being developed to address specific problems in a number of fields of chemistry. These compounds have shown varied coordination modes at metals from across the periodic table, and examples of their application in a number of catalytic processes are emerging as the field matures. PMID- 16474884 TI - Fixation of atmospheric CO2 by a dimeric lanthanum hydroxide complex; assembly of an unusual hexameric carbonate. AB - Exposure of the dinuclear hydroxo complex {[La(tpen)(mu-OH)]2(mu eta1:eta1OTf)}OTf3 to air results in the immediate uptake of atmospheric CO2 affording an unusual hexanuclear lanthanum carbonato complex in which the carbonate anions are ligated in a mu3-eta1:eta1:eta2 and mu3-eta1:eta2:eta2 fashion. PMID- 16474885 TI - Coordinative flexibility in hydrotris(methimazolyl)borate divalent metal compounds. AB - Three divalent closed d shell metal complexes of the hydrotris(methimazolyl)borate ligand, [Ca(mt)2].6H2O, [Ba(mt)2](H2O)2, and Hg4(mt)4Cl4, were synthesized and characterized by single crystal X-ray diffraction; the three structures reveal very different ligand binding modes for each metal ion. PMID- 16474886 TI - Synthesis and characterization of manganese(II) and iron(III) d5 tripodal imidazole complexes. Effect of oxidation state, protonation state and ligand conformation on coordination number and spin state. AB - The 1 : 3 Schiff base condensates of tris(2-aminoethyl)amine (tren) or tris(3 aminopropyl)amine (trpn) with 4-methyl-5-imidazolecarboxaldehyde, H3L1 and H3L2, respectively, were generated in situ and used to prepare complexes with manganese(II) and iron(III). The resultant complexes, [MnH3L1](ClO4)2, [MnH3L1](ClO4)2.EtOH.H2O, [MnH3L2](ClO4)2, [FeH3L1](ClO4)3.1.5(EtOH) and [FeHL1](I3) (0.525)(I)(0.475).2.625H2O, have been characterized by EA, IR, ES MS, variable temperature magnetic susceptibility, X-ray crystallography, and Mossbauer spectroscopy for the iron complexes. The three manganese(II) complexes are high spin with [MnH3L2](ClO4)2 exhibiting coordination number seven while the others are six coordinate. [FeH3L1](ClO4)3.1.5(EtOH) has two iron sites, a seven coordinate and a pseudo seven coordinate site. The complex is high spin at room temperature but exhibits a magnetic moment that decreases with temperature corresponding to conversion of one of the sites to low spin. [FeHL1](I3) (0.525)(I)(0.475).2.625H2O is low spin even at room temperature. In the present complexes the apical nitrogen atom, N(ap), of the tripodal ligand is pyramidal and directed toward the metal atom. The data show that the M-N(ap) distance decreases as the oxidation state of the metal increases, as the number of bound imidazole protons on the ligand increases, and as the number of carbon atoms in the backbone of the ligand (tren vs. trpn) increases. In a limiting sense, short M-N(ap) distances result in high spin seven coordinate mono capped octahedral complexes and long M-N(ap) distances result in low spin six coordinate octahedral complexes. PMID- 16474887 TI - New antitumour active platinum compounds containing carboxylate ligands in trans geometry: synthesis, crystal structure and biological activity. AB - New asymmetric trans-platinum(II) complexes, composed of an isopropylamine, an azole and two carboxylate leaving groups, are presented. The crystal and molecular structures of one of the complexes has been determined and the cytotoxicity and reactivity with 5'-guanosine monophosphate is reported. The complexes show a reduced reactivity, but no decrease in cytotoxic activity compared to their chloro-counterparts. Furthermore the complexes largely overcome cisplatin resistance, they therefore present an interesting class of antitumour active trans-platinum complexes. PMID- 16474888 TI - New route to 1-thia-closo-dodecaborane(11), closo-1-SB11H11, and its halogenation reactions. The effect of the halogen on the dipole moments and the NMR spectra and the importance of spin-orbit coupling for the 11B chemical shifts. AB - Reaction between nido-B10H14 (1) and elemental sulfur in CHCl3 in the presence of Et3N at room temperature, followed by treatment with Et3N.BH3 at 170-190 degrees C, resulted in the isolation of closo-1-SB11H11 (2) in 50% yield. Selected electrophilic halogenation reactions of compound led to the isolation of a series of monohalogenated derivatives of general constitution 12-X-closo-1-SB11H10 (12-X , where X = Cl, Br, and I). The structures of 12-Cl- and 12-I- were determined by an X-ray diffraction analysis and the structures of all compounds were geometry optimised at the RMP2(fc)/6-31G* level. The constitution of all compounds is consistent with the results of mass spectrometry and multinuclear (1H and 11B) spectroscopy complemented by two-dimensional [11B-11B]-COSY and 1H{11B(selective)} NMR measurements. Experimental 11B chemical shifts generally show acceptable agreement with theoretical values calculated by GIAO methods, but spin-orbit coupling must be included for nuclei bearing heavy-atom substituents such as Br or I. The dipole moments determined for the B12-X bonds show similarities to those of aliphatic C-X bonds and confirm unambiguously the B12 - > S dipole moment orientation in the SB11 cage. PMID- 16474889 TI - A chelating beta-diketonate/phenoxide ligand and its coordination behavior toward titanium and scandium. AB - Dibenzoylmethane derivatives with one (L1H2) or both (L2H3, L3H3) benzenes linked at their ortho positions to 4,6-di-tert-butylphenol moieties by two-carbon linkers have been synthesized. The mono-beta-diketone-monophenol ligand L1H2 is metalated by titanium alkoxides to form the homoleptic complex (L1)2Ti and heteroleptic complexes (L1)Ti([OCH2CH2]2NR) (R = H, CH3), and reacts with Cp3Sc to form CpSc(L1). These are the first examples of complexes of a beta-diketonate ligand which is further chelating to a single metal center. Crystallographic analysis of (L1)2Ti indicates that the 10-membered ring allows chelation of the phenoxide with little strain, and both fac and mer geometries are accessible in solution. Protonolysis of the second cyclopentadienyl ring of Cp3Sc appears to take place by an indirect, Cp3Sc-catalyzed pathway. PMID- 16474890 TI - Synthesis and structural characterization of scandium SALEN complexes. AB - A series of heteroleptic scandium SALEN complexes, [(SALEN)Sc(mu-Cl)]2 and (SALEN)Sc[N(SiHMe2)2] is obtained via amine elimination reactions using [Sc(N(i)Pr2)2(mu-Cl)(THF)]2 and Sc[N(SiHMe2)2]3(THF) as metal precursors, respectively. H(2)SALEN ligand precursors comprising H2Salen [(1,2 ethandiyl)bis(nitrilomethylidyne)bis(2,4-di-tert-butyl)phenol], H2Salpren [(2,2 dimethylpropanediyl)bis(nitrilomethylidyne)bis(2,4-di-tert-butyl)phenol], H2Salcyc [(1R,2R)-(-)-1,2-cyclohexanediyl)bis(nitrilomethylidyne)bis(2,4-di-tert butyl)phenol] and H2Salphen [((1S,2S)-(-)-1,2 diphenylethandiyl)bis(nitrilomethylidyne)bis(2,4-di-tert-butyl)phenol] are selected according to solubility and ligand backbone variation ("=N-(R)-N=" bite angle) criteria. Consideration is given to the feasibility of [Cl --> NR2] and [N(SiHMe2)2--> OSiR3] secondary ligand exchange reactions. X-ray crystal structure analyses of donor-free (Salpren)Sc(N(i)Pr2), (R,R) (Salcyc)Sc[N(SiHMe2)2], (Salen)Sc(OSi(t)BuPh2) and (Salphen)Sc(OSiH(t)Bu2) reveal (i) a very short Sc-N bond distance of 2.000(3) A, (ii) weak beta(Si-H)(amido)-Sc agostic interactions and (iii) an exclusive intramolecularly tetradentate and intrinsically bent coordination mode of the SALEN ligands with angle(Ph,Ph) dihedral angles and Sc-[N(2)O(2)] distances in the 124.27(9)-127.7(3) degrees and 0.638(1)-0.688(1) A range, respectively. PMID- 16474892 TI - Intervalence charge transfer in a "chain-like" ruthenium trinuclear assembly based on the bridging ligand 4,7-phenanthrolino-5,6:5',6'-pyrazine (ppz). AB - The IVCT characteristics of the mixed valence forms of the trinuclear complex [{Delta-Ru(bpy)2}2{Delta(t)-Ru(bpy)(mu-ppz)2}]n+ (n = 7, 8; t = trans), and the diastereoisomers (meso and rac) of the dinuclear complex [{Ru(bpy)2}2(mu-ppz)]5+, display a marked dependence on the nuclearity and extent of oxidation of the assemblies. The dinuclear species are classified as borderline localised delocalised mixed valence species while the two mixed valence states of the trinuclear complex exhibit localised behaviour. One-electron oxidation of a terminal Ru centre in the trinuclear case gives rise to a broad, low intensity IVCT band for the +7 mixed valence species which is composed of two underlying Gaussian-shaped bands. The transitions are identified as adjacent and remote IVCT transitions, with the former dominating the intensity of the IVCT manifold. The +8 mixed valence species exhibits a single dominant IVCT band arising from the equivalent IVCT transitions from the central Ru(II) to peripheral Ru(III) centres. PMID- 16474891 TI - Phosphine-ligated induced formation of thallium(I) full Pt3TlPt3 sandwich versus "open-face" TlPt3 sandwich with triangular Pt3(mu2-CO)3(PR3)3 units: synthesis and structural/spectroscopic analysis of triphenylphosphine [(mu3-Tl)Pt3(mu2 CO)3(PPh3)3]+ and its (mu3-AuPPh3)Pt3 analogue. AB - This research constitutes an operational test to assess the influence of platinum attached phosphine ligands in the formation process of "open-face" TlPt3 or "full" Pt3TlPt3 sandwich clusters. Accordingly, the reaction of TlPF6 with triphenylphosphine Pt4(mu2-CO)5(PPh3)4, under essentially identical boundary conditions originally used to prepare (90% yield) the triethylphosphine "full" Pt3TlPt3 sandwich, [(mu6-Tl)Pt6(mu2-CO)6(PEt3)6]+ (3) ([PF6]- salt), from Pt4(mu2 CO)5(PEt3)4 was carried out to see whether it would likewise afford the unknown triphenylphosphine Pt3TlPt3 sandwich analogue of or whether the change of phosphine ligands from sterically smaller, more basic PEt3 to PPh3 would cause the product to be the corresponding unknown triphenylphosphine "open-face" TlPt3 sandwich that would geometrically resemble the known bulky tricyclohexylphosphine [(mu3-Tl)Pt3(mu2-CO)3(PCy3)3]+ sandwich (2a). Both the structure and composition of the resulting "open-face" sandwich product, [(mu3-Tl)Pt3(mu2-CO)3(PPh3)3]+ (1a) ([PF6]- salt), were unequivocally established from a low-temperature CCD X ray crystallographic determination. The calculated Pt/Tl atom ratio (3/1) of 75%/25% is in excellent agreement with that of 72(3)%/28(5)% obtained from energy resolved measurements on a single crystal with a scanning electron microscope. Crystals (80% yield) of the orange-red were characterized by solid-state/solution IR and variable temperature 205Tl and 31P{1H} NMR spectra; the 31P{1H} spectra provide convincing evidence that is exhibiting dynamic behavior at room temperature in CDCl3 solution. The corresponding new "open-face" (mu3-AuPPh3)Pt3 sandwich, [(mu3-AuPPh3)Pt3(mu2-CO)3(PPh3)3]+ (1b) ([PF6]- salt), was quantitatively obtained from by reaction with AuPPh3Cl and spectroscopically characterized by IR and 31P{1H} NMR spectra. A comparative geometrical evaluation of the observed steric dispositions of the platinum-attached PR3 ligands in the "open-face" (mu3-Tl)Pt3 sandwiches of (with PPh3) and the known (with PCy3) and in the known "full" Pt3TlPt3 sandwich of (with PEt3) along with the considerably different observed steric dispositions of the PR(3) ligands in the known "open face" (mu3-AuPCy3)Pt3 sandwich of (with PCy3) and in the known "full" Pt3AuPt3 sandwich of (with PPh(3)) has been performed. The results clearly indicate that, in contradistinction to the known triphenylphosphine Pt3AuPt3 sandwich of , PPh3 and bulkier PCy3 ligands of Pt3(mu2-CO)3(PR3)3 units are sterically too large to form "full" Pt3TlPt3 sandwiches. In other words, the nature of the thallium(I) sandwich-product in these reactions is sterically controlled by size effects of the phosphine ligands. Comparative examination of bridging carbonyl IR frequencies of and with those of closely related "open-face" and "full" sandwiches provides better insight concerning the relative electrophilic capacities of Tl+, Au+, and [AuPR3]+ components in forming sandwich adducts with Pt3(mu2-CO)3(PR3)3 nucleophiles. PMID- 16474893 TI - Coinage metal complexes of tris(pyrazolyl)methanide [C(3,5-Me2pz)3]-: kappa3 coordination vs. backbone functionalisation. AB - Tris(pyrazolyl)methanides, [C(3,5-R2pz)3]-, contain an unassociated tetrahedral carbanionic centre in the bridgehead position. In addition to nitrogen donor centres for transition metal coordination, an accessible reactive site for further manipulations is available in the backbone of the ligand. The coordination variability of the ambidental C-/N ligand [C(3,5-Me2pz)3]- was elucidated by investigating its coinage metal complexes. Two principle coordination modes were found for complexes of general formula [LMPR3] (with M = Cu(I), Ag(I), Au(I); L =[C(3,5-Me2pz)3]-; R = Ph, OMe). While for Cu(I) (2,3) and Ag(I) (4) complexes the anionic ligand acts as a face-capping, six electron N3 donor, gold(I) (5) is coordinated by the bridging carbanion yielding a two coordinate Au(I) complex comprising a covalent Au-C bond. The complexes featuring the kappa3-coordinated N3-donor ligand were investigated by 31P CP (MAS) NMR in the solid state. PMID- 16474894 TI - Equilibrium and NMR studies on GdIII, YIII, CuII and ZnII complexes of various DTPA-N,N''-bis(amide) ligands. Kinetic stabilities of the gadolinium(III) complexes. AB - Three DTPA-derivative ligands, the non-substituted DTPA-bis(amide) (L(0)), the mono-substituted DTPA-bis(n-butylamide) (L(1)) and the di-substituted DTPA bis[bis(n-butylamide)] (L(2)) were synthesized. The stability constants of their Gd3+ complexes (GdL) have been determined by pH-potentiometry with the use of EDTA or DTPA as competing ligands. The endogenous Cu2+ and Zn2+ ions form ML, MHL and M(2)L species. For the complexes CuL(0) and CuL(1) the dissociation of the amide hydrogens (CuLH(-1)) has also been detected. The stability constants of complexes formed with Gd3+, Cu2+ and Zn2+ increase with an increase in the number of butyl substituents in the order ML(0) < ML(1) < ML(2). NMR studies of the diamagnetic YL(0) show the presence of four diastereomers formed by changing the chirality of the terminal nitrogens of their enantiomers. At 323 K, the enantiomerization process, involving the racemization of central nitrogen, falls into the fast exchange range. By the assignment and interpretation of 1H and 13C NMR spectra, the fractions of the diastereomers were found to be equal at pH = 5.8 for YL(0). The kinetic stabilities of GdL(0), GdL(1) and GdL(2) have been characterized by the rates of the exchange reactions occurring between the complexes and Eu3+, Cu2+ or Zn2+. The rates of reaction with Eu3+ are independent of the [Eu3+] and increase with increasing [H+], indicating the rate determining role of the proton assisted dissociation of complexes. The rates of reaction with Cu2+ and Zn2+ increase with rising metal ion concentration, which shows that the exchange can take place with direct attack of Cu2+ or Zn2+ on the complex, via the formation of a dinuclear intermediate. The rates of the proton, Cu2+ and Zn2+ assisted dissociation of Gd3+ complexes decrease with increasing number of the n butyl substituents, which is presumably the result of steric hindrance hampering the formation or dissociation of the intermediates. The kinetic stabilities of GdL(0) and GdL(1) at pH = 7.4, [Cu2+] = 1 x 10(-6) M and [Zn(2+)] = 1 x 10(-5) M are similar to that of Gd(DTPA)2-, while the complex GdL2 possesses a much higher kinetic stability. PMID- 16474895 TI - Molecular modelling of Jahn-Teller distortions in Cu(II)N6 complexes: elongations, compressions and the pathways in between. AB - Ligand Field Molecular Mechanics (LFMM) parameters have been optimised for six coordinate Cu(II) complexes containing amine, pyridine, imidazole and pyrazine donors. As found in previous LFMM applications, the new parameters automatically generate distorted structures with the magnitudes of the Jahn-Teller elongations in good agreement with experiment. Here, we explore the rest of the potential energy surface. The introduction of axial strain drives the LFMM structures via rhombic geometries to the compressed structure, the latter corresponding to the saddle point between successive elongation axes. Calculated barrier heights between compressed and elongated geometries also agree well with available experimental data. In every case bar one, the LFMM predicts that the crystallographically observed elongation axis corresponds to the overall lowest energy well. The structural predictions are confirmed by independent density functional theory (DFT) optimisations. LFMM calculations on bis(2,5 pyrazolylpyridine)copper complexes display a smooth variation in structure as a function of pyrazolyl substituent from elongated for R = H through to fully compressed for R = (t)Bu. This behaviour is driven by the steric interactions with the ground state varying smoothly as a linear combination of {d(x2-y2)}1 and {d(z2)}1. PMID- 16474896 TI - Synthesis, characterization and application of Ni(tta)2.tmeda to MOCVD of nickel oxide thin films. AB - A novel nickel beta-diketonate adduct, Ni(tta)2.tmeda, has been synthesized using 2-thenoyltrifluoroacetone as the beta-diketonate and N,N,N,'N' tetramethylethylendiamine as the Lewis base. It has been characterized by elemental analyses, IR, 1H NMR, 13C NMR spectroscopy and single-crystal X-ray diffraction studies. Physical and thermal properties of Ni(tta)2.tmeda precursor have been also extensively investigated. Its efficacy as a metal-organic chemical vapour deposition (MOCVD) precursor for the growth of nickel oxide films has been fully tested by applying it to the deposition of NiO films on quartz substrate. NiO thin films have been characterized by X-ray diffraction (XRD), scanning electron microscopy and UV spectroscopy. PMID- 16474897 TI - Anti-HIV I/II activity and molecular cloning of a novel mannose/sialic acid binding lectin from rhizome of Polygonatum cyrtonema Hua. AB - The anti-human immunodeficiency virus (HIV) I/II activity of a mannose and sialic acid binding lectin isolated from rhizomes of Polygonatum cyrtonema Hua was elucidated by comparing its HIV infection inhibitory activity in MT-4 and CEM cells with that of other mannose-binding lectins (MBLs). The anti-HIV activity of Polygonatum cyrtonema Hua lectin (PCL) was 10- to 100-fold more potent than other tested MBLs, but without significant cytotoxicity towards MT-4 or CEM cells. To amplify cDNA of PCL by 3'/5'-rapid amplification of cDNA ends (RACE), the 30 amino acids of N-terminal were determined by sequencing and the degenerate oligonucleotide primers were designed. The full-length cDNA of PCL contained 693 bp with an open reading frame encoding a precursor protein of 160 amino acid residues, consisting of a 28-residue signal peptide, a 22-residue C-terminal cleavage peptide and a 110-residue mature polypeptide which contained three tandemly arranged subdomains with an obvious sequence homology to the monocot MBL. However, only one active mannose-binding site (QDNVY) was found in subdomain I of PCL, that of subdomain II and III changed to HNNVY and PDNVY, respectively. There was no intron in PCL, which was in good agreement with other monocot MBLs. Molecular modeling of PCL indicated that its three-dimensional structure resembles that of the snowdrop agglutinin. By docking, an active sialic acid binding site was found in PCL. The instabilization of translation initiation region (TIR) in mRNA of PCL benefits its high expression in rhizomes. PMID- 16474898 TI - Characterization of cheW genes of Leptospira interrogans and their effects in Escherichia coli. AB - The motility and chemotaxis systems are critical for the virulence of leptospires. In this study, the phylogenetic profiles method was used to predict the interaction of chemotaxis proteins. It was shown that CheW1 links to CheA1, CheY, CheB and CheW2; CheW3 links to CheA2, MCP (LA2426), CheB3 and CheD1; and CheW2 links only to CheW1. The similarity analysis demonstrated that CheW2 of Leptospira interrogans strain Lai had poor homology with CheW of Escherichia coli in the region of residues 30-50. In order to verify the function of these proteins, the putative cheW genes were cloned into pQE31 vector and expressed in wild-type E. coli strain RP437 or cheW defective strain RP4606. The swarming results indicated that CheW1 and CheW3 could restore swarming of RP4606 while CheW2 could not. Overexpression of CheW1 and CheW3 in RP437 inhibited the swarming of RP437, whereas the inhibitory effect of CheW2 was much lower. Therefore, we presumed that CheW1 and CheW3 might have the function of CheW while CheW2 does not. The existence of multiple copies of chemotaxis homologue genes suggested that L. interrogans strain Lai might have a more complex chemosensory pathway. PMID- 16474899 TI - Regulated production of mature insulin in rat hepatoma cells: insulin production is up-regulated by dexamethasone and down-regulated by insulin. AB - We engineered an artificial beta cell line that produces an up-regulation of insulin in response to dexamethasone, and a down-regulation in response to insulin. A regulatory secretion system was devised by placing proinsulin cDNA containing genetically engineered furin endoprotease cleavage sites and a regulatory promoter for phosphoenolpyruvate carboxykinase (PEPCK), and an insulin expressing retrovirus vector (pN-PEPCK-mINS) was constructed and transfected into Hepa1-6 cells. The levels of insulin in culture medium and expression of insulin gene was estimated by radioimmunoassay and reverse transcription-polymerase chain reaction (RT-PCR), respectively. The clone (Hepa1-6/INS21), which secreted the highest level of insulin (10.79 microIU/106 cells per day), was selected for the regulation experiment. Compared with the non-treated Hepa1-6/INS21 cells, insulin production was augmented 3.6-fold by the addition of 10-7 M of dexamethasone. Addition of exogenous insulin to the culture medium decreased insulin mRNA expression remarkably on RT-PCR results, while dexamethasone increased insulin gene expression at the transcriptional level. The data indicated that genetically engineered Hepa1-6 cells could synthesize process and secrete insulin in a physiological manner. PMID- 16474900 TI - Roles of cortactin, an actin polymerization mediator, in cell endocytosis. AB - Cortactin, an actin-binding protein and a substrate of Src, is encoded by the EMS1 oncogene. Cortactin is known to activate Arp2/3 complex-mediated actin polymerization and interact with dynamin, a large GTPase and proline rich domain containing protein. Transferrin endocytosis was significantly reduced in cells by knock-down of cortactin expression as well as in vivo introduction of cortactin immunoreagents. Cortactin-dynamin interaction displayed morphologically dynamic co-distribution with a change in the endocytosis level in cells treated with an actin depolymerization reagent, cytochalasin D. In an in vitro beads assay, a branched actin network was recruited onto dynamin-coated beads in a cortactin Src homology domain 3 (SH3)-dependent manner. In addition, cortactin was found to function in the late stage of clathrin coated vesicle formation. Taken together, cortactin is required for optimal clathrin mediated endocytosis in a dynamin directed manner. PMID- 16474901 TI - Transient decrease of light-harvesting complex II phosphorylation level by hypoosmotic shock in dark-adapted Dunaliella salina. AB - This study investigated the regulation of major light harvesting chlorophyll a/b protein (LHCII) phosphorylation by hypoosmotic shock in dark-adapted Dunaliella salina cells. When the external NaCl concentration decreased in darkness, D. salina LHCII phosphorylation levels transiently dropped within 20 min and then restored gradually to basal levels. The transient decrease in LHCII phosphorylation levels was insensitive to NaF, a phosphatase inhibitor. Inhibition of intracellular ATP production by addition of an uncoupler or an ATP synthase inhibitor increased LHCII phosphorylation levels in D. salina cells exposed to hypoosmotic shock. Taken together, these results indicate that hypoosmotic shock inhibits the LHCII phosphorylation process. The related mechanism and physiological significance are discussed. PMID- 16474902 TI - Identification of melanoma-specific peptide epitopes by HLA-A2.1-restricted cytotoxic T lymphocytes. AB - HLA-A2.1-associated peptides, extracted from human melanoma cells, were used to study epitopes for melanoma-specific HLA-A2.1-restricted cytotoxic T lymphocytes (CTLs) by epitope reconstitution, active peptide sequence characterization and synthetic peptide verification. CTL were generated from tumor-involved nodes by in vitro stimulation, initially with autologous melanoma cells and subsequently with allogeneic HLA-A2.1 positive melanoma cells. The CTLs could lyse autologous and allogeneic HLA-A2.1 positive melanomas, but not HLA-A2.1 negative melanomas or HLA-A2.1 positive non-melanomas. The lysis of melanomas could be inhibited by anti-CD3, anti-HLA class I and anti-HLA-A2.1 monoclonal antibodies. HLA-A2.1 molecules were purified from detergent-solubilized human melanoma cells by immunoaffinity column chromatography and further fractionated by reversed phase high performance liquid chromatography. The fractions were assessed for their ability to reconstitute melanoma-specific epitopes with HLA-A2.1 positive antigen processing mutant T2 cells. Three reconstitution peaks were observed in lactate dehydrogenase release assay. Mass spectrometry and ion-exchange high performance liquid chromatography analysis were used to identify peptide epitopes. Peptides with a mass-to-charge ratio of 948 usually consist of nine amino acid residues. The data from reconstitution experiments confirmed that the synthetic peptides contained epitopes and that the peptides associated with HLA-A2.1 and recognized by melanoma-specific CTL were present in these different melanoma cells. These peptides could be potentially exploited in novel peptide-based antitumor vaccines in immunotherapy for CTL. PMID- 16474903 TI - Regulatory peptides modulate adhesion of polymorphonuclear leukocytes to bronchial epithelial cells through regulation of interleukins, ICAM-1 and NF kappaB/IkappaB. AB - A complex network of regulatory neuropeptides controls airway inflammation reaction, in which airway epithelial cells adhering to and activating leukocytes is a critical step. To study the effect of intrapulmonary regulatory peptides on adhesion of polymorphonuclear leukocytes (PMNs) to bronchial epithelial cells (BECs) and its mechanism, several regulatory peptides including vasoactive intestinal peptide (VIP), epidermal growth factor (EGF), endothelin-1 (ET-1) and calcitonin gene-related peptide (CGRP), were investigated. The results demonstrated that VIP and EGF showed inhibitory effects both on the secretion of IL-1, IL-8 and the adhesion of PMNs to BECs, whereas ET-1 and CGRP had the opposite effect. Anti-intercellular adhesion molecule-1 (ICAM-1) antibody could block the adhesion of PMNs to ozone-stressed BECs. Using immunocytochemistry and reverse transcription-polymerase chain reaction (RT-PCR), it was shown that VIP and EGF down-regulated the expression of ICAM-1 in BECs, while ET-1 and CGRP up regulated ICAM-1 expression. NF-kappaB inhibitor MG132 blocked ICAM-1 expression induced by ET-1 and CGRP. Furthermore, in electric mobility shift assay (EMSA), VIP and EGF restrained the binding activity of NF-kappaB to the NF-kappaB binding site within the ICAM-1 promoter in ozone-stressed BECs, while CGRP and ET-1 promoted this binding activity. IkappaB degradation was consistent with NF-kappaB activation. These observations indicate that VIP and EGF inhibit inflammation, while ET-1 and CGRP enhance the inflammation reaction. PMID- 16474904 TI - Evaluation of antibody responses elicited by immunization of mice with a pneumococcal antigen genetically fused to murine HSP70 and murine interleukin-4. AB - The heat shock (stress) protein HSP70 has been shown to be a potent stimulator of cellular immune responses. In order to determine whether HSP70 has the ability to stimulate antibody responses, we constructed and expressed fusion proteins consisting of murine HSP70 or murine interleukin (IL)-4 covalently linked to a pneumococcal cell wall-associated protein antigen designated PpmA. Immunization of mice with the PpmA-HSP70 fusion protein (PpmA-70) failed to elicit an increased PpmA-specific serum antibody response. In contrast, mice immunized with PpmA fused to IL-4 (PpmA-IL4), or PpmA fused to both IL-4 and HSP70 (PpmA-IL4-70) fusion proteins elicited high levels of PpmA-specific antibody responses. These data suggest that HSP70 has a limited capacity to stimulate immune responses to heterologous antigens in vivo. PMID- 16474905 TI - Kir6.2DeltaC26 channel traffics to plasma membrane by constitutive exocytosis. AB - Adenosine triphosphate (ATP)-sensitive K+ (KATP) channels regulate many cellular functions by coupling the metabolic state of the cell to the changes in membrane potential. Truncation of C-terminal 26 amino acid residues of Kir6.2 protein (Kir6.2DeltaC26) deletes its endoplasmic reticulum retention signal, allowing functional expression of Kir6.2 in the absence of sulfonylurea receptor subunit. pEGFP-Kir6.2DeltaC26 and pKir6.2DeltaC26-IRES2-EGFP expression plasmids were constructed and transfected into HEK293 cells. We identified that Kir6.2DeltaC26 was localized on the plasma membrane and trafficked to the plasmalemma by means of constitutive exocytosis of Kir6.2DeltaC26 transport vesicles, using epi fluorescence and total internal reflection fluorescence microscopy. Our electrophysiological data showed that Kir6.2DeltaC26 alone expressed KATP currents, whereas EGFP-Kir6.2DeltaC26 fusion protein displayed no KATP channel activity. PMID- 16474906 TI - Novel cold-adaptive Penicillium strain FS010 secreting thermo-labile xylanase isolated from Yellow Sea. AB - A novel cold-adaptive xylanolytic Penicillium strain FS010 was isolated from Yellow sea sediments. The marine fungus grew well from 4 to 20 degrees; a lower (0 degrees) or higher (37 degrees) temperature limits its growth. The strain was identified as Penicillium chrysogenum. Compared with mesophilic P. chrysogenum, the cold-adaptive fungus secreted the cold-active xylanase (XYL) showing high hydrolytic activities at low temperature (2-15 degrees) and high sensitivity to high temperature (>50 degrees). The XYL gene was isolated from the cold-adaptive P. chrysogenum FS010 and designated as xyl. The deduced amino acid sequence of the protein encoded by xyl showed high homology with the sequence of glycoside hydrolase family 10. The gene was subcloned into an expression vector pGEX-4T-1 and the encoded protein was overexpressed as a fusion protein with glutathione-S transferase in Escherichia coli BL21. The expression product was purified and subjected to enzymatic characterization. The optimal temperature and pH for recombinant XYL was 25 degrees and 5.5, respectively. Recombinant XYL showed nearly 80% of its maximal activity at 4 degrees and was active in the pH range 3.0-9.5. PMID- 16474907 TI - Accuracy of preoperative imaging of hepatic tumors with helical computed tomography. AB - BACKGROUND: The accuracy of preoperative computed tomography (CT) scans in the era of modern imaging techniques with helical, high-resolution CT has not been adequately assessed. We reviewed the data from our departmental prospective database with the hypothesis that intraoperative ultrasonography (IOUS) still detects more hepatic tumors than are evident on preoperative helical CT scans. METHODS: All patients who underwent surgical resection and/or radiofrequency ablation of primary or metastatic hepatic tumors between January 2001 and July 2002 were included in the review. All patients had preoperative helical CT imaging followed by hepatic IOUS. The number of malignant lesions and evidence of local disease identified by the preoperative CT scan versus IOUS and surgical exploration were compared. RESULTS: In this time period, 250 patients underwent surgical resection and/or radiofrequency ablation of hepatic tumors. In 67 (27%) of these patients, IOUS identified more hepatic tumors than were seen on preoperative helical CT scan. In eight patients (3%), CT underestimated local extension of the disease into the diaphragm. The incidence of inaccurate preoperative prediction of the extent of disease increased significantly with a greater number of hepatic tumors. CONCLUSIONS: IOUS identified additional hepatic tumors in 27% of patients who underwent hepatic resection after state-of-the-art preoperative CT imaging. This study provides evidence that IOUS remains an essential part of the complete assessment of hepatic malignancies in patients who receive surgical treatment. PMID- 16474908 TI - Significance of papillary lesions at percutaneous breast biopsy. AB - BACKGROUND: The management of nonpalpable papillary lesions found in specimens obtained by percutaneous breast biopsy is controversial. We reviewed the treatment of patients found to have papillary lesions by stereotactic, sonographic, or fine-needle aspiration breast biopsy to identify indications for surgical excision. METHODS: Consecutive patients with intraductal papilloma, atypical papilloma/papilloma with atypical ductal hyperplasia, papillary neoplasm, and papillomatosis according to percutaneous breast biopsy were identified from radiology records. The charts were reviewed to identify patients who had subsequent surgical excision, and the pathologic findings were correlated with the biopsy method and indications for surgery. RESULTS: Papillary lesions were found in 120 biopsy samples from 109 patients. Malignancy was found at operation in 19 (24%) of 80 lesions that underwent surgical excision: 12 (63%) were ductal carcinoma-in-situ, 4 (21%) were infiltrating ductal carcinoma, 2 (11%) were infiltrating papillary carcinoma, and 1 (5%) was intracystic papillary carcinoma. Malignancy was found in 9 (30%) of 30 fine-needle biopsy papillary lesions, 6 (35%) of 17 core biopsy papillary lesions, and 4 (12%) of 33 stereotactic biopsy papillary lesions. Malignancy was missed significantly less frequently with stereotactic biopsy (P<.05). CONCLUSIONS: Malignancy is frequently found at surgical excision for papillary lesions found on percutaneous breast biopsy. Malignancy is missed significantly less frequently with stereotactic biopsy. PMID- 16474909 TI - Utility of preoperative [(18)]f fluorodeoxyglucose-positron emission tomography scanning in high-risk melanoma patients. AB - BACKGROUND: [(18)]F Fluorodeoxyglucose-positron emission tomography (PET) scanning provides functional imaging based on glucose uptake by tumors. Melanoma is a glucose-avid malignancy, and preoperative PET scanning in melanoma patients has the potential to guide appropriate treatment. METHODS: We performed a prospective trial to evaluate the clinical utility of whole-body fluorine 18 labeled deoxyglucose-PET scanning used in addition to standard imaging (contrast enhanced computed tomographic [CT] imaging of the chest, abdomen, and pelvis) in preoperative stage IIC (T4N0M0), III (any T, N1 to N3, M0), and IV (any T, any N, M1) melanoma patients. Pathologic or clinical follow-up within 4 to 6 months of the imaging studies was used to determine the accuracy of preoperative PET and CT scan findings. RESULTS: Preoperative imaging findings led to a change in clinical management in 36 (35%) of 103 patients. In 32 (89%) of these patients, the information was accurate. Findings on PET scan alone (14 of 36; 39%) or in combination with CT (20 of 36; 56%) resulted in a treatment change in most patients (34 of 36; 94%). The most common decision was to cancel the operation (19 of 36; 53%). PET scanning was more sensitive than CT scanning in detecting occult disease (68% vs. 48%; P=.05), but both tests were highly specific (92% vs. 95%; P=.7, PET vs. CT). CONCLUSIONS: PET scanning facilitates the appropriate management of high-risk melanoma patients being considered for operative intervention. PET imaging in addition to CT scanning should be strongly considered before operation in patients at high risk for occult metastatic disease. PMID- 16474910 TI - Anaplastic thyroid carcinoma: biology, pathogenesis, prognostic factors, and treatment approaches. AB - BACKGROUND: Anaplastic thyroid carcinoma (ATC) is one of the most aggressive solid tumors known to affect humans and carries a dismal prognosis. Our primary aim was to review its epidemiology, biology, risk factors, and prognostic indicators. We also reviewed the individual and combined roles of surgery, radiotherapy, chemotherapy, and newer therapeutic options in the management of ATC. METHODS: An extensive literature review was conducted to include all published reports on ATC. The changing trends in the management of anaplastic thyroid cancer were analyzed to summarize the current practice of management of ATC. RESULTS: Although ATC is rare, there has been a decline in its incidence worldwide. ATC accounts for more than half of the 1200 deaths per year attributed to thyroid cancer. Long-term survivors are rare, with >75% and 50% of patients harboring cervical nodal disease and metastatic disease, respectively, at presentation. ATC can arise de novo or from preexisting well-differentiated thyroid cancer. Surgical management has shifted from tracheostomy only for palliation to curative resection when possible. Tracheostomy is performed for impending obstruction rather than for prophylaxis. Radiotherapy has evolved from postoperative administration only to preoperative treatment, combining preoperative and postoperative treatment and using higher doses, along with hyperfractionating and accelerating dose schedules. Chemotherapy has changed from monotherapy to combination therapy, and newer drugs such as paclitaxel show promise. Similarly, novel angiogenesis-inhibiting agents are currently being used, with early reports of some benefit. CONCLUSIONS: Despite multimodality approaches, ATC still carries a dismal prognosis. This should provoke innovative strategies beyond conventional methods to tackle this uniformly lethal disease. PMID- 16474911 TI - A pathologic assessment of adequate margin status in breast-conserving therapy. AB - BACKGROUND: The definition of a clear margin in breast-conserving therapy is uncertain. The purpose of this study was to correlate the tumor-margin distance of the excision specimen with the presence of residual tumor at reoperation. We also analyzed predictors of compromised margins and of residual disease. METHODS: All patients who underwent breast-conserving therapy for invasive disease from 1999 to 2003 were reviewed. Pathologic characteristics and the precise tumor distance from the radial margin were recorded. A radial margin was compromised if invasive or (ductal) in situ carcinoma was <5 mm from the margin. RESULTS: Of the 612 patients who underwent breast conservation, 211 (34%) had compromised margins, and 39 had undetermined margins. Of the 161 patients who had a reoperation for compromised margins, 87 (54%) had residual disease. Residual disease after reoperation was present in 58% (56 of 96), 56% (9 of 16), and 45% (22 of 49) of those with tumor-margin distances <1 mm, > or =1 and <2 mm, and > or =2 and <5 mm, respectively. There was a progressive decline in residual disease for each millimeter until a rate of 22% for tumor-margin distances of > or =4 mm and <5 mm was reached. Pathologic size (P = .004), an extensive intraductal component (P = .002), referral from a symptomatic rather than a population-based screening program (P = .02), and the absence of a preoperative diagnosis by core biopsy (P < .0001) were predictive of compromised margins. Only young age (<45 years) was predictive of finding residual disease on reoperation (P = .02). CONCLUSIONS: A total of 45% of patients who had tumor 2 to 5 mm from the radial margin had residual disease on reoperation. Our results support a policy of requiring a 5-mm margin in patients undergoing breast-conserving therapy for invasive disease. PMID- 16474912 TI - Profile of plasma angiogenic factors before and after hepatectomy for colorectal cancer liver metastases. AB - BACKGROUND: Circulating angiogenic factors in patients with colorectal cancer liver metastases may promote tumor growth and contribute to liver regeneration after partial hepatectomy. METHODS: We analyzed blood samples from 26 patients with colorectal cancer liver metastases before and after liver resection and used samples from 20 healthy controls as a reference. Plasma levels of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), and hepatocyte growth factor (HGF) were measured, and levels were correlated with recurrence. RESULTS: The median preoperative levels of all four factors were significantly higher and more variable in colorectal cancer liver metastasis patients than in controls. HGF and bFGF levels increased significantly 3 days and 1 month after hepatectomy, respectively, and returned to near preoperative levels at 3 months. Postoperative VEGF and EGF levels remained relatively stably increased over 3 months. After a median follow up of 19 months, 10 patients (42%) experienced recurrence. Higher preoperative VEGF and HGF levels correlated with subsequent recurrence (P = .018 and .021, respectively), and a preoperative adjusted total value of all four factors accurately identified patients at low, moderate, and high risk of recurrence (P = .034). Patients who experienced disease recurrence also had relatively higher bFGF levels 3 months after operation (P = .035). CONCLUSIONS: Plasma angiogenic factors are increased in patients with colorectal cancer liver metastases and remain increased at least 3 months after partial hepatectomy. Measurement of certain factors before and after hepatic resection can predict recurrence. Targeted biological agents may counteract the tumor-promoting effects of these circulating factors on subclinical disease. PMID- 16474913 TI - Academic impairment is the most frequent complication of neurofibromatosis type-1 (NF1) in children. AB - Neurofibromatosis type-1 (NF1) is a common genetic disorder associated with a variety of medical complications, cognitive impairments, and behavioral problems. One hundred and sixteen patients with NF1 (62 males, 54 females; mean age 12.4 years) and 80 typically developing children of the same ages (46 males, 34 females; mean age 11.5 years) were studied in terms of complications and learning impairment (one or more grade repetitions or school exclusion). Seventy of 116 patients had significant learning impairment. Classical complications were present in 53 patients including the three. PMID- 16474914 TI - Likelihood ratio tests in behavioral genetics: problems and solutions. AB - The likelihood ratio test of nested models for family data plays an important role in the assessment of genetic and environmental influences on the variation in traits. The test is routinely based on the assumption that the test statistic follows a chi-square distribution under the null, with the number of restricted parameters as degrees of freedom. However, tests of variance components constrained to be non-negative correspond to tests of parameters on the boundary of the parameter space. In this situation the standard test procedure provides too large p-values and the use of the Akaike Information Criterion (AIC) or the Bayesian Information Criterion (BIC) for model selection is problematic. Focusing on the classical ACE twin model for univariate traits, we adapt existing theory to show that the asymptotic distribution for the likelihood ratio statistic is a mixture of chi-square distributions, and we derive the mixing probabilities. We conclude that when testing the AE or the CE model against the ACE model, the p values obtained from using the chi(2)(1 df) as the reference distribution should be halved. When the E model is tested against the ACE model, a mixture of chi(2)(0 df), chi(2)(1 df) and chi(2)(2 df) should be used as the reference distribution, and we provide a simple formula to compute the mixing probabilities. Similar results for tests of the AE, DE and E models against the ADE model are also derived. Failing to use the appropriate reference distribution can lead to invalid conclusions. PMID- 16474915 TI - Bioengineering and systems biology. PMID- 16474917 TI - Interdisciplinary BME education: a clinical preceptorship course for undergraduate bioengineering students. PMID- 16474919 TI - A finite element model for ultrafast laser-lamellar keratoplasty. AB - A biomechanical model of the human cornea is employed in a finite element formulation for simulating the effects of Ultrafast Laser-Lamellar Keratoplasty. Several computer simulations were conducted to study curvature changes of the central corneal zone under various physiological and surgical factors. These factors included the combined effect of corneal flap and residual stromal bed thickness on corneal curvature; the effect of the shape of the lenticle on the surgical procedure outcomes and the effect of flap thickness on stress distribution in the cornea. The results were validated by comparing computed refractive power changes with clinical results. The effect of flap thickness on the amount of central flattening indicates that for flap thickness values 28% over the corneal thickness, central corneal flattening decreases. Moreover, the change in corneal curvature induced by subtraction of a plano-convex lenticle under a uniform flap, naturally imply a smaller change in the structure of the anterior layers of the cornea, but a bigger deformation in the structure of the posterior layers that are left behind the resection of the lenticle. In addition, the model also verified that the corneal curvature increased peripherally with simultaneous thinning centrally after subtraction of corneal tissue. This result shows that not only the treated zone is affected by the surgery, indicating the important role of the biomechanical response of the corneal tissue to refractive surgery, which is unaccounted for in current ablation algorithms. The results illustrate the potentialities of finite element modeling as an aid to the surgeon in evaluating variables. PMID- 16474921 TI - A review of the Northern Ecosystem Initiative in Arctic Canada: facilitating arctic ecosystem research through traditional and novel approaches. AB - The Canadian Arctic is undergoing considerable social and environmental change. Anthropogenic stressors on this sensitive environment include climate change, contaminants, resource extraction, tourism and increasing human populations. The Northern Ecosystem Initiative (NEI) is a program aimed at supporting the sustainability of northern communities, and at improving our understanding of how northern ecosystems respond to these environmental stressors. A key element of the NEI is to establish partnerships between all levels of government, non governmental environmental agencies, and northern residents. The NEI is an important source of financial support critical for social and environmental scientists as well as northern residents and their community and regional organizations. Initiated in 1998, the NEI has supported numerous northern scientific and capacity-building projects, and has evaluated the information gained from this work to refine and focus its future support to address key information gaps and northern needs. PMID- 16474916 TI - A probabilistic model of glenohumeral external rotation strength for healthy normals and rotator cuff tear cases. AB - The reigning paradigm of musculoskeletal modeling is to construct deterministic models from parameters of an "average" subject and make predictions for muscle forces and joint torques with this model. This approach is limited because it does not perform well for outliers, and it does not model the effects of population parameter variability. The purpose of this study was to simulate variability in musculoskeletal parameters on glenohumeral external rotation strength in healthy normals, and in rotator cuff tear case using a Monte Carlo model. The goal was to determine if variability in musculoskeletal parameters could quantifiably explain variability in glenohumeral external rotation strength. Multivariate Gamma distributions for musculoskeletal architecture and moment arm were constructed from empirical data. Gamma distributions of measured joint strength were constructed. Parameters were sampled from the distributions and input to the model to predict muscle forces and joint torques. The model predicted measured joint torques for healthy normals, subjects with supraspinatus tears, and subjects with infraspinatus-supraspinatus tears with small error. Muscle forces for the three conditions were predicted and compared. Variability in measured torques can be explained by differences in parameter variability. PMID- 16474922 TI - Application of CCME Water Quality Index to monitor water quality: a case study of the Mackenzie River Basin, Canada. AB - All six ecosystem initiatives evolved from many years of federal, provincial, First Nation, local government and community attention to the stresses on sensitive habitats and species, air and water quality, and the consequent threats to community livability. This paper assesses water quality aspect for the ecosystem initiatives and employs newly developed Canadian Council of Ministers of the Environment Water Quality Index (CCME WQI) which provides a convenient mean of summarizing complex water quality data that can be easily understood by the public, water distributors, planners, managers and policy makers. The CCME WQI incorporates three elements: Scope - the number of water quality parameters (variables) not meeting water quality objectives (F(1)); Frequency - the number of times the objectives are not met (F(2)); and Amplitude. the extent to which the objectives are not met (F(3)). The index produces a number between 0 (worst) to 100 (best) to reflect the water quality. This study evaluates water quality of the Mackenzie - Great Bear sub-basin by employing two modes of objective functions (threshold values): one based on the CCME water quality guidelines and the other based on site-specific values that were determined by the statistical analysis of the historical data base. Results suggest that the water quality of the Mackenzie-Great Bear sub-basin is impacted by high turbidity and total (mostly particulate) trace metals due to high suspended sediment loads during the open water season. Comments are also provided on water quality and human health issues in the Mackenzie basin based on the findings and the usefulness of CCME water quality guidelines and site specific values. PMID- 16474923 TI - Targeted sustainable development: 15 years of government and community intervention on the St. Lawrence River. AB - From 1988 to 2003, the St. Lawrence Action Plan, a Canada - Quebec cooperation agreement on the St. Lawrence River, helped to mobilize a still-growing number of stakeholders in the conservation and protection of this great river and generated tangible results in a number of areas of intervention. The successes enjoyed in the area of industrial, agricultural and urban clean-up, in protecting plant and animal species and their habitats, and in acquiring new knowledge on the state and trends of the ecosystem reflect the sustained efforts of government and non government partners over the course of those 15 years. The committed involvement of local communities provides assurance of the sustainable development of this vast ecosystem. PMID- 16474925 TI - Biomechanics of the deltoideus. AB - The objective of this study was to determine the direction of the migration engendered by the middle deltoideus on the upper end of the humerus. Eleven patients suffering from shoulder pathology underwent an MRI examination (3 mm thick slices). From these MRI slices, 3D reconstructions were obtained for each patient by using a manual data capture system (SliceOmatic((R))). From this geometry, a mechanical model of the deltoideus was produced, taking into account the contacts between the latter and the following anatomical parts: supraspinatus, infraspinatus and humeral head. For the 11 shoulders, we have obtained a deltoideus showing a global resultant oriented upwards. There was, however, a component oriented downwards (at the level of the humeral head), its intensity being 40-80% less than the component oriented upwards (at the level of the deltoideus V). It is important to note that this study is valid only in the initial degrees of lateral elevation. The deltoideus is an elevator muscle of the humeral head in the glenoid, presenting nevertheless a component oriented downwards. The deltoideus would, therefore, intervene to recenter the shoulder during an abduction movement. PMID- 16474924 TI - Ligamentoplasty of the forearm interosseous membrane using the semitendinosus tendon: anatomical study and surgical procedure. AB - Total longitudinal disruptions of the interosseous membrane can allow proximal radius migration and are seen in Essex-Lopresti lesions. We propose an original technique of ligamentoplasty using the semitendinosus tendon. The graft corresponds to the forearm rotation axis for an optimized isometry and longitudinal stabilization. Our ligamentoplasty technique was performed on ten fresh frozen right forearms. We successively assessed the innocuousness, efficiency and resistance of the ligamentoplasty. The ligamentoplasty induced neither passive limitation of pronation-supination nor neurovascular lesions. It prevented from radius proximal migration. The mean load to failure was 28 kg at both ulnar and radial sides of the graft. Our technique is original for the type and position of the graft. It seems safe, efficient and resistant enough for in vivo procedures. This technique decreases longitudinal loads on the radius. It should be indicated in patients with Essex-Lopresti syndrome, in association with radial head internal fixation or arthroplasty. PMID- 16474926 TI - Subclavian vein obstruction with collateral flow through the umbilical and hepatic veins. AB - Upper venous return to the heart can be obstructed by any number of pathologies and often leads to formation of collateral pathways around the obstruction. This case report describes a thrombotic obstruction of the left subclavian and brachiocephalic veins and a collateral pathway involving recanalization of the umbilical vein. Even more rare and unique is the anastomosis of the recanalized umbilical vein to the middle and left hepatic vein rather than the portal vein. The anatomy of the expected collateral pathways and the anastomotic vascular anomaly is described in detail, including a discussion of the embryological development of the system. PMID- 16474927 TI - Nerve fiber analysis for the lingual nerve of the human adult subjects. AB - The neuro-motor control of the human tongue musculature had not been investigated in detail. This study identified first that the lingual nerve should play the neuro-motor control of some lingual muscles. Six en bloc samples (12 sides), including the tissues from the skull base to the hyoid bone, and three whole tongues were obtained from adult human cadavers. The former samples were used for the study of nerve fiber analysis of the lingual nerve with the aid of binocular stereomicroscope, and the latter samples were used for histological study by serial section method. On nerve fiber analysis of the lingual nerve from the trigeminal ganglion to the tongue musculature, we found that the motor- root of the trigeminal nerve gave off its supply to the lingual nerve and traveled into the lingual nerve, and branched to the superior and the inferior longitudinal muscles. On histological study, it was revealed that in the anterior part of the tongue the superior and the inferior longitudinal muscles surrounded the other lingual musculature and combined with the sub-mucosal connective tissues closely like the cutaneous muscle, for example, the facial muscles. The lingual nerve entered the inner side of the space between the genioglossus and the inferior longitudinal muscles with the lingual artery. These findings suggested that the superior and the inferior longitudinal muscles should be innervated by the motor fibers traveled into the lingual nerve from the motor root of the trigeminal nerve, and do not originate from the myotome originating in occipital somites but branchial muscles. PMID- 16474928 TI - Phenotypic alterations and survival of monocytes following infection by human herpesvirus-6. AB - Freshly isolated monocytes rapidly undergo physiological changes in vitro, resulting in programmed cell death (apoptosis). Activation of monocytes, which promotes differentiation into macrophages, is known to inhibit apoptotic processes. In the present study, we report that human herpesvirus-6 (HHV-6) prevents monocytes from undergoing spontaneous apoptosis during the first 72 hours of culture. Furthermore, significant alterations in cell-surface phenotype were observed after 72 hours of infection with HHV-6. HHV-6-infected monocyte cultures have considerably reduced levels of CD14, CD64 (FcgammaRI) and HLA-DR antigen on their surface, while CD32 (FcgammaRII) expression is unaffected. On the basis of these results, we hypothesize that HHV-6 promotes monocytes survival and causes phenotypic modifications that could favor immune evasion and ensure its persistence within the infected host. PMID- 16474929 TI - Damage of PS II during senescence of Spirodela polyrrhiza explants under long-day conditions and its prevention by 6-benzyladenine. AB - The chlorophyll a (Chl a) fluorescence technique was applied to investigate damage of PS II during senescence of excised half-fronds in Spirodela polyrrhiza P143. The green explants showed a typical Chl a fluorescence transient, OJIP. After cultivation of explants under long-day conditions for 8 days, all the J, I, and P steps disappeared, but a clear K band, an indication of senescence, was observed. JIP-test showed that at this time point, the photosynthetic performance index (PI) dropped to zero and the active reaction center (RC) per leaf cross section (RC/CS) declined to 18%. As the oxygen-evolving complex (OEC) and the chlorophyll content all remained above 42%, it is proposed that the decline in RC contributes more to the appearance of the K band. Supplementation of 6 benzyladenine (6-BA) into the medium at the beginning of cultivation caused dramatic increase in PI, OEC, RC/CS, and chlorophyll content, and at any time before the 8th day reversed the senescence process of the explants. When 6-BA was added after 8 days of cultivation, the PI did not increase anymore, RC/CS and OEC were maintained at 22% and above 40%, respectively, and chlorophyll content decreased continuously further. These data support a view that the decline in RC is crucial for initiation of the irreversible senescence phase of explants cultivated under long-day conditions. PMID- 16474930 TI - Insight into the basis of root growth in Arabidopsis thaliana provided by a simple mathematical model. AB - Plant organ growth changes under genetic and environmental influences can be observed as altered cell proliferation and volume growth. The two aspects are mutually dependent and intricately related. For comprehensive growth analysis, it is necessary to specify the relationship quantitatively. Here, we develop a simple mathematical model for this purpose. Our model assumes that the biological activity of a given organ is proportional to the cell number of the organ and is allocated into three aspects: cell proliferation, volume growth, and organ maintenance. We analyzed the growth of primary roots of Arabidopsis thaliana (L.) Heynh. in one tetraploid and four diploid strains using this model. The analysis determined various growth parameters, such as specific cost coefficients of cell proliferation and volume growth for each strain. The results provide insight into the basis of interstrain variations and ploidy effects in root growth. PMID- 16474931 TI - A puzzle in DNA biophysics. AB - In this issue, Lee et al. report the experimental temperature-dependence of the unzipping force for two natural DNA sequences. For both sequences, the curves show an anomaly at temperatures around 40 degrees C. In this brief contribution, we stress that the anomaly is not easily explained within the established theoretical models for the biophysics of DNA. As this puzzle questions our basic understanding of DNA, it must be resolved, most likely by a combination of additional experiments and new theoretical work. PMID- 16474932 TI - MRI features of tibiofibular joint dislocation. PMID- 16474933 TI - Association between the MLH1 gene and longevity. AB - Perturbations in genomic stability result in cancer, a reduced life span, and premature aging. MLH1 is a mismatch repair enzyme that acts to maintain genomic stability, and a loss of MLH1 increases cancer incidence and apoptosis resistance, which suggests a link between MLH1 and longevity. We found here that MLH1 is associated with longevity by comparing a centenarian group with a control group. Our data indicate a critical role for MLH1 in longevity. PMID- 16474934 TI - Interferon gamma polymorphisms and their interaction with smoking are associated with lung function. AB - Interactions between genetic and environmental determinants are likely to be important in the pathogenesis of chronic obstructive pulmonary disease. We hypothesized that interferon gamma (IFNG) single nucleotide polymorphisms (SNPs) and their interaction with smoking are associated with the rate of decline or level of lung function in smokers. We studied four SNPs in IFNG in 585 non Hispanic whites (NHW) who had the fastest (n =280) or the slowest (n=305) decline of FEV(1)% predicted selected from among continuous smokers followed for 5 years in the NHLBI Lung Health Study. We also studied 1061 NHW with the lowest (n=530) or the highest (n=531) baseline lung function at the beginning of the LHS. Two SNPs were associated with baseline levels of lung function and the p values were 0.008 for +2197T/C in a dominant model and 0.002 for +5171A/G in a recessive model. However, after adjusting for confounding factors, only +5171A/G was still significant (p=0.001 for the recessive model). In addition, there was a significant genotype and smoking interaction with p=0.006 for the +5171A/G (GG vs.GA + AA) for the baseline lung function. When comparing individuals with GG versus individuals with AG + AA for low lung function, the adjusted odds ratios decreased significantly as pack-years increased. No association was found in the rate of decline study. There was an association between IFNG genotype and baseline of lung function and this association was modified by cigarette smoking. PMID- 16474935 TI - Role of PKC in regulation of Fos and TH expression after naloxone induced morphine withdrawal in the heart. AB - We previously demonstrated that morphine withdrawal induced hyperactivity of the heart by activation of noradrenergic pathways innervating the left and right ventricle, as evaluated by noradrenaline (NA) turnover and Fos expression. The present study was designed to investigate the role of protein kinase C (PKC) in this process, by estimating whether pharmacological inhibition of PKC would attenuate morphine withdrawal induced Fos expression and changes in tyrosine hydroxylase (TH) immunoreactivity levels and NA turnover in the left and right ventricle. Dependence on morphine was induced on day 8 by an injection of naloxone. Morphine withdrawal induced Fos expression and increased TH levels and NA turnover in the right and left ventricle. Infusion of calphostin C, a selective PKC inhibitor, did not modify the morphine withdrawal-induced increase in NA turnover and TH levels. However, this inhibitor produced a reduction in the morphine withdrawal-induced Fos expression. The results of the present study provide new information on the mechanisms that underlie morphine withdrawal induced up-regulation of Fos expression in the heart and suggest that TH is not a target of PKC during morphine withdrawal at heart levels. PMID- 16474937 TI - Overcorrection during treatment of pectus deformities with DCC orthoses: experience in 17 cases. AB - Treatment of pectus carinatum and pectus excavatum with dynamic chest compressor (DCC) orthoses have been reported by Haje and others. The goal of this study was to demonstrate that overcorrection during orthotic treatment of children and adolescents with pectus deformities can occur and requires medical attention. Of 3,028 children and adolescents with pectus deformities, observed between 1977 and October 2005, 1,824 were prescribed treatment with DCC orthoses and, after a few months of treatment, some overcorrection was noted in 30 patients. Of the patients who received orthoses, 738 had a minimum follow-up of 1 year and 17 of these, 2 with pectus excavatum and 15 with pectus carinatum, presented overcorrection and were studied. The dynamic remodeling method (DCC orthoses + exercises) was applied. The procedures, adopted according to each patient's needs, were: decreasing the time of orthosis wear and/or the tightening of the screws, introducing a second orthosis, and improving the prescribed exercises and/or encouraging the patient to perform them more intensively. The therapy was successful in all patients, and the result was maintained in one case of pectus excavatum followed up until adulthood. It was concluded that overcorrection during DCC orthosis wear can occur and that careful medical follow-up is necessary if this complication is to be successfully reversed. PMID- 16474936 TI - Impending fractures in giant cell tumours of the distal femur: incidence and outcome. AB - Giant cell tumours are rare bone tumours that are characteristically benign but locally aggressive, most frequently occurring in the distal femur with pathological fractures being common. This paper investigates relationships between tumour size and cortical breach on initial X-rays and subsequent treatment. The X-rays of 54 patients with distal femoral giant cell tumours were reviewed. The volumes of the tumour, distal femur and a ratio between the two parameters were estimated. The presence of a cortical breach, discrete fracture and Campanacci grade was recorded. X-rays revealed intact cortical rim in 20 patients (37%), cortical breach in 22 patients (41%) and discrete fracture in 12 patients (22%). There was a significant difference in the ratio of tumour volume to distal femoral volume between the discrete fracture group and the cortical breach group. No significant differences in rates of local recurrence were demonstrated. Extended curettage was effective for intact and cortical breach groups; however, patients in the fracture group often required radical treatment. PMID- 16474939 TI - The effect of ultrasound on the healing of muscle-pediculated bone graft in scaphoid non-union. AB - The use of pedicled vascularised bone grafts from the distal radius makes it possible to transfer bone with a preserved circulation and viable osteoclasts and osteoblasts. Experiments performed at the basic science level has provided substantial evidence that low-intensity ultrasound can accelerate and augment the fracture healing process. Only an adequate double-blind trial comparing treatment by ultrasound stimulation in patients treated by similar surgical techniques can provide evidence of the true effect of ultrasound. This paper describes the results of such a trial. From 1999 to 2004, 21 fractures of the scaphoid with established non-union treated with vascularised pedicle bone graft were selected for inclusion in a double-blind trial. All patients were males, with an average age of 26.7 years (range 17-42 years) and an average interval between injury and surgery of 38.4 months (range 3 months-10 years). Low-intensity ultrasound was delivered using a TheraMed 101-B bone-growth stimulator (30 mW/cm2, 20 min/day), which was modified to accomplish double-blinding. These modifications did not affect the designated active units. The placebo units were adjusted to give no ultrasound signal output across the transducer. Externally, all units appeared identical but were marked with individual code numbers. Patients were randomly allocated to either an active or placebo stimulation. Follow-up averaged 2.3 years (range 1-4 years). All patients achieved fracture union (active and placebo groups), but compared with the placebo device (11 patients), the active device (ten patients) accelerated healing by 38 days (56+/-3.2 days compared with 94+/ 4.8 days, p<0.0001, analysis of variance). PMID- 16474938 TI - Percutaneous CT-guided curettage of osteoid osteoma with histological confirmation: a retrospective study and review of the literature. AB - Osteoid osteoma is a benign bone tumour usually occurring in young individuals (10-30 years). It presents with intense pain (typically nocturnal), which can be alleviated by salicylates. Treatment consists of surgical excision or destroying the nidus and it is curative. In the past, surgery was performed in an "open" fashion and the nidus had to be removed with a bone block. This extensive type of surgery could be associated with some rates of both failure and complication. There is growing evidence to suggest that percutaneous CT-guided removal or destruction of the nidus is a good alternative and it is indeed gaining worldwide popularity. We present a series of 18 consecutive patients with osteoid osteoma of the pelvis, femur, and tibia, treated percutaneously under CT guidance. Removal of the nidus was performed using a 4.5-mm cannulated drill and a cannulated curette of our own design. Tissue samples for histological evaluation were obtained in the same way. The mean follow-up time was 29 months. Sixteen patients were initially cured. The procedure had to be repeated in two patients and was eventually successful (primary and secondary success rates 88 and 100% respectively). The diagnosis was histologically confirmed in 14 cases out of 18 (77%). In four cases no histological confirmation of osteoid osteoma could be achieved. There were only two minor complications, one case of femoral neuropraxia and one case of skin abrasion. Percutaneous CT-guided removal seems to be efficient and safe for the treatment of osteoid osteoma. The use of a cannulated drill and a cannulated curette facilitates efficient removal of the tumour and procurement of tissue for diagnosis. PMID- 16474940 TI - Successful treatment with linezolid and rifampicin of meningitis due to methicillin-resistant Staphylococcus epidermidis refractory to vancomycin treatment. PMID- 16474941 TI - MiniX-STR multiplex system population study in Japan and application to degraded DNA analysis. AB - We sought to evaluate a more effective system for analyzing X-chromosomal short tandem repeats (X-STRs) in highly degraded DNA. To generate smaller amplicon lengths, we designed new polymerase chain reaction (PCR) primers for DXS7423, DXS6789, DXS101, GATA31E08, DXS8378, DXS7133, DXS7424, and GATA165B12 at X-linked short tandem repeat (STR) loci, devising two miniX-multiplex PCR systems. Among 333 Japanese individuals, these X-linked loci were detected in amplification products ranging in length from 76 to 169 bp, and statistical analyses of the eight loci indicated a high usefulness for the Japanese forensic practice. Results of tests on highly degraded DNA indicated the miniX-STR multiplex strategies to be an effective system for analyzing degraded DNA. We conclude that analysis by the current miniX-STR multiplex systems offers high effectiveness for personal identification from degraded DNA samples. PMID- 16474942 TI - Endoscopic surgery on the thoracolumbar junction of the spine. AB - The thoracolumbar junction is the section of the truncal spine most often affected by injuries. Acute instability with structural damage to the anterior load-bearing spinal column and post-traumatic deformity represents the most frequent indications for surgery. In the past few years, endoscopic techniques for these indications have partially superseded the open procedures, which are associated with high access morbidity. The particular position of this section of the spine, which lies in the transition area between the thoracic and abdominal cavities, makes it necessary in most cases to partially detach the diaphragm endoscopically in order to expose the surgical site, and this also provides access to the retroperitoneal section of the thoracolumbar junction. A now standardised operating technique, instruments and implants specially developed for the endoscopic procedure, from angle stable plate and screw implants to endoscopically implantable vertebral body replacements, have gradually opened up the entire spectrum of anterior spine surgery to endoscopic techniques. PMID- 16474943 TI - Clinical predictors of lumbar provocation discography: a study of clinical predictors of lumbar provocation discography. AB - Discography is the only available method of directly challenging the discs for pain sensitivity. However, it is invasive, expensive and there is a debate about its clinical value. There is a need to identify clinical signs that may indicate the need for a discography examination. Pain centralization (retreat of referred pain to towards the spinal midline) has been associated with positive discography and possesses specificity between 80 and 100% depending on levels of disablement and psychosocial distress. Less than half of positive discography patients reports centralization and there is a need to identify other potentially valuable predictors. This study estimated the diagnostic accuracy of clinical variables in relation to provocation discography. In a blinded, prospective reference standard design in a private interventional radiology clinic, chronic low back patients received a detailed clinical examination followed by discography. Patients were typically disabled with high levels of psychosocial distress. Logistic regression modelling identified potentially useful clinical variables, and sensitivity, specificity and likelihood ratios were calculated for promising specific variables and combinations of variables. Of 118 consecutive discography cases, 107 had unconfounded data on discography results. History of persistent pain between acute episodes, a significant loss of extension and a subjective report of 'vulnerability' in what is termed as the 'neutral zone' had specificities of 83-92% and likelihood ratios between 2.0 and 4.1. Two combinations of variables were highly specific to positive discography, but no suitable screening test useful for ruling out positive discography was established. Three clinical variables have modest predictive power in relation to lumbar discography results and two combinations of variables were highly specific to positive discography. PMID- 16474944 TI - Epithelioid sarcoma in the thoracic spine. AB - Epithelioid sarcoma is a rare and highly malignant soft tissue tumor that is commonly found in the extremities and rarely in the trunk area. This malignant tumor often mimics granuloma or nodular fasciitis, which causes a delay in establishing the diagnosis. This type of cancer has a high recurrence rate. Surgical treatment requires wide radical resection. The objective of this case report is to highlight the unique location of a rare neoplasm and to illustrate the relentless course of epithelioid sarcoma despite initial radical resection. A 14-year-old boy was admitted to our facility with a soft tissue mass on the right lower thoracic spine. The large tumor mass had deeply penetrated into the muscles, infiltrated the neuroforamen of T9-T10 level, and compressed the dural sac. Immunohistological study of the biopsy was highly consistent with an epithelioid sarcoma. Wide excision of the mass, laminectomy and spine fusion with instrumentation was performed. The patient received chemotherapy and irradiation. The first recurrence of the neoplasm was seen as a contralateral metastasis 21 months after the resection. On the last follow-up, 3 years postoperatively, the patient was in a good general condition. However, further progression of the sarcoma had to be recognized. Our case encompasses multiple features that represent negative prognostic factors. Initial wide excision of the neoplasm and adjuvant therapy including chemotherapy and irradiation seem to slow down the relentless course of epithelioid sarcoma in the trunk. PMID- 16474945 TI - Effects of interleukin-1 beta, interleukin-6, and tumor necrosis factor on sensitivity of dorsal root ganglion and peripheral receptive fields in rats. AB - This study was designed to characterize the effects of low doses (0.5-5 ng) of pro-inflammatory cytokines, interleukin-1 beta (IL-1beta), interleukin-6 (IL-6), and tumor necrosis factor (TNF), on the neural activity of dorsal root ganglion (DRG) in rats. The purpose of this study was to examine the effects of cytokines (IL-1beta, IL-6, and TNF) on the somatosensory neural response of DRG. The release of inflammatory cytokines by an injured disc may play a critical role in pain production at nerve endings, axons, and nerve cell bodies. Herniated disc tissue has been shown to release IL-1beta, IL-6, TNF, and other algesic chemicals. Their effects on nerve endings in disc and adjacent tissue may lead to low back pain and their effects on dorsal root axons and ganglia may lead to sciatica. Exposed lumbar DRGs were investigated by electrophysiologic techniques. Sham (mineral oil), control (carrier solution), or IL-1beta, IL-6, or TNF at doses of 0.5, 1, and 5 ng were applied over the DRG. Baseline discharge rates as well as mechanosensitivity of the DRG and peripheral receptive fields were evaluated over 30 min. Applications of IL-1beta at 1 ng resulted in an increase in the discharge rate, 5 ng resulted in an increased mechanosensitivity of the DRG in group II units. Similarly, after 1 ng TNF applications, group II units also showed an increase in mechanosensitivity of DRG and peripheral receptive fields. At low doses IL-1beta and TNF sensitization of receptive fields were observed. The responses observed in the group II units indicate that a sub population of afferent units might have long-term effects modifying the sensory input to the central nervous system. This study provides added evidence to the role of cytokines in modulating afferent activity following inflammation. PMID- 16474947 TI - Giant cervicothoracic extradural arachnoid cyst: case report. AB - The pathogenesis, etiology, and treatment of the spinal arachnoid cyst have not been well established because of its rarity. A 57-year-old male was presented with spastic quadriparesis predominantly on the left side. His radiological examination showed widening of the cervical spinal canal and left neural foramina due to a cerebrospinal fluid-filled extradural cyst that extended from C2 to T2 level. The cyst was located left anterolaterally, compressing the spinal cord. Through a C4-T2 laminotomy, the cyst was excised totally and the dural defect was repaired. Several features of the reported case, such as cyst size, location, and clinical features make it extremely unusual. The case is discussed in light of the relevant literature. PMID- 16474946 TI - Glucocorticosteroid-induced spinal osteoporosis: scientific update on pathophysiology and treatment. AB - Glucocorticosteroid-induced spinal osteoporosis (GIOP) is the most frequent of all secondary types of osteoporosis. The understanding of the pathophysiology of glucocorticoid (GC) induced bone loss is of crucial importance for appropriate treatment and prevention of debilitating fractures that occur predominantly in the spine. GIOP results from depressed bone formation due to lower activity and higher death rate of osteoblasts on the one hand, and from increase bone resorption due to prolonged lifespan of osteoclasts on the other. In addition, calcium/phosphate metabolism may be disturbed through GC effects on gut, kidney, parathyroid glands and gonads. Therefore, therapeutic agents aim at restoring balanced bone cell activity by directly decreasing apoptosis rate of osteoblasts (e.g., cyclical parathyroid hormone) or by increasing apoptosis rate of osteoclasts (e.g., bisphosphonates). Other therapeutical efforts aim at maintaining/restoring calcium/phosphate homeostasis: improving intestinal calcium absorption (using calcium supplementation, vitamin D and derivates) and avoiding increased urinary calcium loss (using thiazides) prevent or counteract a secondary hyperparthyroidism. Bisphosphonates, particularly the aminobisphosphonates risedronate and alendronate, have been shown to protect patients on GCs from (further) bone loss to reduce vertebral fracture risk. Calcitonin may be of interest in situation where bisphosphonates are contraindicated or not applicable and in cases where acute pain due to vertebral fracture has to be manage. The intermittent administration of 1-34-parathormone may be an appealing treatment alternative, based on its documented anabolic effects on bone resulting from the reduction of osteoblastic apoptosis. Calcium and vitamin D should be a systematic adjunctive measure to any drug treatment for GIOP. Based on currently available evidence, fluoride, androgens, estrogens (opposed or unopposed) cannot be recommended for the prevention and treatment of GIOP. However, substitution of gonadal hormones may be indicated if GC-induced hypogonadism is present and leads to clinical symptoms. Data using the SERM raloxifene to treat or prevent GIOP are lacking, as are data using the promising bone anabolic agent strontium ranelate. Kyphoplasty performed in appropriately selected osteoporotic patients with painful vertebral fractures is a promising addition to current medical treatment. PMID- 16474949 TI - CT-guided obturator nerve block via the posterior approach. AB - OBJECTIVE: To describe the technique of obturator nerve block under CT guidance via the posterior approach, and to evaluate the efficacy of the procedure in the short-term and mid-term relief of chronic hip pain. DESIGN AND PATIENTS: Consecutive patients referred for obturator nerve block were prospectively enrolled in this study. Under CT-guidance, via a posterior approach through the pelvis, local anaesthetic and steroid were infiltrated around the obturator nerve using a 22G spinal needle. Fifty-one patients (19 male, 32 female), mean age 54 years, with hip pain refractory to conventional therapy underwent the procedure. Visual Analogue Scale pain scores were recorded before the procedure and at 30 minutes, 24 hours, 1 week and 3 months thereafter. RESULTS: Pain scores within 30 minutes showed a decrease from a mean +/- SD score of 8.41+/-1.22 pre-procedure to 2.86+/-2.1, p<0.001. At 24 hours, the mean pain score was 2.06+/-1.76, a decrease of 76% from pre-procedural score, p<0.001. Sustained pain relief at 1 week and 3 months was attained in 92% (mean pain score 2.41+/-2.2, p<0.001) and 82% (mean pain score 3.80+/-2.94, p<0.001) of cases respectively. Follow-up data was complete for all 51 patients. No serious side-effects were reported. CONCLUSIONS: In patients with hip pain refractory to conventional pain control measures, CT-guided obturator nerve block can provide relief from pain in the short to medium term. The posterior approach offers safe, reliable and effective access to the nerve, in a procedure which is well-tolerated by the patient. PMID- 16474948 TI - Influence of body size on urinary stone composition in men and women. AB - A larger body size has been shown to be associated with increased excretion of urinary lithogenic solutes, and an increased risk of nephrolithiasis has been reported in overweight patients. However, the type of stones produced in these subjects has not been ascertained. Based on a large series of calculi, we examined the relationship between body size and the composition of stones, in order to assess which type of stone is predominantly favoured by overweight. Among 18,845 consecutive calculi referred to our laboratory, 2,100 came from adults with recorded body height and weight. Excluding calculi from patients with diabetes mellitus, as well as struvite and cystine stones, the study material consisted of 1,931 calcium or uric acid calculi. All calculi were analysed by infrared spectroscopy and categorized according to their main component. Body mass index (BMI) values were stratified as normal BMI (< 25 kg/m2), overweight (BMI 25-29.9) or obese (BMI > or = 30). Overall, 27.1% of male and 19.6% of female stone formers were overweight, and 8.4 and 13.5% were obese, respectively. In males, the proportion of calcium stones was lower in overweight and obese groups than in normal BMI group, whereas the proportion of uric acid stones gradually increased with BMI, from 7.1% in normal BMI to 28.7% in obese subjects (P<0.0001). The same was true in females, with a proportion of uric acid stones rising from 6.1% in normal BMI to 17.1% in obese patients (P=0.003). In addition, the proportion of uric acid stones markedly rose with age in both genders (P<0.0001). The average BMI value was significantly higher in uric acid stone formers aged < 60 years than in all other groups, whereas it did not differ from other groups in those aged > or = 60 years. Stepwise regression analysis identified BMI and age as significant, independent covariates associated with the risk of uric acid stones. Our data provide evidence that overweight is associated with a high proportion of uric acid stones in patients less than 60 years of age, whereas beyond this limit, advancing age is the main risk factor. PMID- 16474950 TI - Characterization of product capture resin during microbial cultivations. AB - Various bioactive small molecules produced by microbial cultivation are degraded in the culture broth or may repress the formation of additional product. The inclusion of hydrophobic adsorber resin beads to capture these products in situ and remove them from the culture broth can reduce or prevent this degradation and repression. These product capture beads are often subjected to a dynamic and stressful microenvironment for a long cultivation time, affecting their physical structure and performance. Impact and collision forces can result in the fracturing of these beads into smaller pieces, which are difficult to recover at the end of a cultivation run. Various contaminating compounds may also bind in a non-specific manner to these beads, reducing the binding capacity of the resin for the product of interest (fouling). This study characterizes resin bead binding capacity (to monitor bead fouling), and resin bead volume distributions (to monitor bead fracture) for an XAD-16 adsorber resin used to capture epothilone produced during myxobacterial cultivations. Resin fouling was found to reduce the product binding capacity of the adsorber resin by 25-50%. Additionally, the degree of resin bead fracture was found to be dependent on the cultivation length and the impeller rotation rate. Microbial cultivations and harvesting processes should be designed in such a way to minimize bead fragmentation and fouling during cultivation to maximize the amount of resin and associated product harvested at the end of a run. PMID- 16474951 TI - FTIR spectroscopy as a potential tool to analyse structural modifications during morphogenesis of Candida albicans. AB - Candida albicans is a polymorphic organism that grows under certain conditions as blastospores, hyphae or pseudohyphae. The potentials of FTIR spectroscopy for assessing structural differences in C. albicans blastospores and hyphae were investigated. The main observed differences were localised in the polysaccharide (950-1,185 cm(-1)), protein (1,480-1,720 cm(-1)), and the fatty acids (2,840 3,000 cm(-1)) regions. Quantitative evaluation of differences between hyphae and blastospores by curve-fitting of these regions indicate that these modifications could be due to both changes in structure and content of components of the cell wall such as beta-glucans, mannoproteins, and lipids. Furthermore, glycogen consumption could be involved during hyphae elongation. Thus, FTIR spectroscopy can be an interesting tool to investigate differences in structure and in content between blastospores and hyphae. We also demonstrate through this study that differentiation of C. albicans clinical strains using hyphae is feasible, as this has been previously shown with blastospores. This preliminary work on identification of C. albicans using hyphae is a prelude to a larger clinical study for early typing within 7 h from a pure culture. PMID- 16474952 TI - ISPa20 advances the individual evolution of Pseudomonas aeruginosa clone C subclone C13 strains isolated from cystic fibrosis patients by insertional mutagenesis and genomic rearrangements. AB - Pseudomonas aeruginosa clone C strains, which chronically colonize the lungs of cystic fibrosis patients reorganize their genome structure. In this study, a novel member of the IS3 subfamily of IS elements, ISPa20, was detected which was specific for clone C subclone C13 strains. ISPa20, which was present in high copy number, mediated events of genomic reorganization. ISPa20 was inserted into P. aeruginosa backbone genes leading to adaptation to the cystic fibrosis lung habitat and into DNA acquired through horizontal gene transfer. Further on, large chromosomal inversions were mediated by ISPa20. In contrast to strains of other subclonal linages high rates of genomic rearrangements of subclone C13 strains were observed in vitro. The acquisition of mobile elements by P. aeruginosa clone C strains in the lungs of cystic fibrosis patients supports the chronic colonization by insertional mutagenesis and chromosome restructuring leading to microevolution within clone C that reflects macroevolution observed on the species level. PMID- 16474953 TI - Collagen I:III ratio in canine heterologous bladder acellular matrix grafts. AB - The objective of this study was to investigate the limitations of a heterologous bladder acellular matrix graft (BAMG) and the influence of the collagen ratio on functional regeneration in a large animal model. Ten female dogs underwent partial cystectomy; eight received BAMG (two homologous; six heterologous) and two partial cystectomy only. A cystometry was performed prior to surgery and 7 months postoperatively when all animals underwent sacral root stimulation. Tissue specimens were studied by histologic and immunohistochemical techniques and for collagen types. At month 7, all animals survived and bladder capacity in the grafted animals was increased. All grafts demonstrated all components of a normal bladder wall. Nerves were seen with the density decreasing with distance from the anastomosis. The BAMG processing and follow-up demonstrated no changes in the homologous tissue, whereas in the heterologous tissue, the amount of collagen changed with the processing during implantation. None of these heterologous specimens demonstrated a similar collagen ratio to the hosts'. The homologous BAMG undergoes more complete regeneration. In the heterologous BAMG, collagen seems not to be replaced. The amounts and ratio of collagen types I and III seem to influence smooth muscle regeneration. PMID- 16474956 TI - Comment on "Changes in acid-base balance following bolus infusion of 20% albumin solution in humans" by Bruegger et al. PMID- 16474954 TI - Organ dysfunction during sepsis. AB - BACKGROUND: Multiple organ dysfunction syndrome is the commonest reason for sepsis-associated mortality. DISCUSSION: In the 40 years since it was first described understanding of its pathophysiology has improved, and novel methodologies for monitoring and severity of illness scoring have emerged. These, together with the development of systematic strategies for managing organ dysfunction in sepsis, and potentially effective new therapeutic interventions, should assist in reducing sepsis-associated mortality. CONCLUSION: These historical developments are discussed, and the reader is directed to these references for further guidance. PMID- 16474957 TI - Blockade and enhancement of glutamate receptor responses in Xenopus oocytes by methylated arsenicals. AB - Pentavalent and trivalent organoarsenic compounds belong to the major metabolites of inorganic arsenicals detected in humans. Recently, the question was raised whether the organic arsenicals represent metabolites of a detoxification process or methylated species with deleterious biological effects. In this study, the effects of trivalent arsenite (AsO(3) (3-); iA(III)), the pentavalent organoarsenic compounds monomethylarsonic acid (CH(3)AsO(OH)(2); MMA(V)) and dimethylarsinic acid ((CH(3))(2)AsO(OH); DMA(V)) and the trivalent compounds monomethylarsonous acid (CH(3)As(OH)(2), MMA(III)) and dimethylarsinous acid ((CH(3))(2)As(OH); DMA(III)) were tested on glutamate receptors and on voltage operated potassium and sodium channels heterologously expressed in Xenopus oocytes. Membrane currents of ion channels were measured by conventional two electrode voltage-clamp techniques. The effects of arsenite were tested in concentrations of 1-1,000 micromol/l and the organic arsenical compounds were tested in concentrations of 0.1-100 micromol/l. We found no significant effects on voltage-operated ion channels; however, the arsenicals exert different effects on glutamate receptors. While MMA(V) and MMA(III) significantly enhanced ion currents through N-methyl-D: -aspartate (NMDA) receptor ion channels with threshold concentrations <10 micromol/l, DMA(V) and DMA(III) significantly reduced NMDA-receptor mediated responses with threshold concentrations <0.1 micromol/l; iA(III) had no effects on glutamate receptors of the NMDA type. MMA(III) and DMA(V) significantly reduced ion currents through alpha-amino-3 hydroxy-5-methylisoxazole-4-propionic acid (AMPA)-receptor ion channels with threshold concentrations <10 micromol/l (MMA(III)) and <1 micromol/l (DMA(V)). MMA(V) and iA(III) had no significant effects on glutamate receptors of the AMPA type. The effects of MMA(V), MMA(III), DMA(V) and DMA(III )on glutamate receptors point to a neurotoxic potential of these substances. PMID- 16474958 TI - Cytotoxicity of dental composite (co)monomers and the amalgam component Hg(2+) in human gingival fibroblasts. AB - Unpolymerized resin (co)monomers or mercury (Hg) can be released from restorative dental materials (e.g. composites and amalgam). They can diffuse into the tooth pulp or the gingiva. They can also reach the gingiva and organs by the circulating blood after the uptake from swallowed saliva. The cytotoxicity of dental composite components hydroxyethylmethacrylate (HEMA), triethyleneglycoldimethacrylate (TEGDMA), urethanedimethacrylate (UDMA), and bisglycidylmethacrylate (Bis-GMA) as well as the amalgam component Hg(2+) (as HgCl(2)) and methyl mercury chloride (MeHgCl) was investigated on human gingival fibroblasts (HGFs) at two time intervals. To test the cytotoxicity of substances, the bromodeoxyuridine (BrdU) assay and the lactate dehydrogenase (LDH) assay were used. The test substances were added in various concentrations and cells were incubated for 24 or 48 h. The EC(50) values were obtained as half-maximum-effect concentrations from fitted curves. Following EC(50) values were found [BrdU: mean (mmol/l); SEM in parentheses; n=12]: (24 h/48 h) HEMA 8.860 (0.440)/6.600(0.630), TEGDMA 1.810(0.130)/1.220(0.130), UDMA 0.120(0.010)/0.140(0.010), BisGMA 0.060(0.004)/0.040(0.002), HgCl(2) 0.015(0.001)/0.050(0.006), and MeHgCl 0.004(0.001)/0.005(0.001). Following EC(50) values were found [LDH: mean (mmol/l); SEM in parentheses; n=12]: (24 h/48 h) HEMA 9.490(0.300)/7.890(1.230), TEGDMA 2.300(0.470)/1.950(0.310), UDMA 0.200(0.007)/0.100(0.007), BisGMA 0.070(0.005)/0.100(0.002), and MeHgCl 0.014(0.006)/0.010(0.003). In both assays, the following range of increased toxicity was found for composite components (24 and 48 h): HEMA < TEGDMA < UDMA < BisGMA. In both assays, MeHgCl was the most toxic substance. In the BrdU assay, Hg(2+) was about fourfold less toxic than MeHgCl but Hg(2+) was about fourfold more toxic than BisGMA. In the BrdU test, a significantly (P<0.05) decreased toxicity was observed for Hg(2+) at 48 h, compared to the 24 h Hg(2+)-exposure. A time depending decreased toxicity was observed only for Hg(2+) which can then reach the toxic level of BisGMA. PMID- 16474959 TI - Ethanol hyperpolarizes mitochondrial membrane potential and increases mitochondrial fraction in cultured mouse myocardial cells. AB - Cultured mouse heart-derived myocardial and non-muscle cells were exposed to ethanol, stained with cell-permeant fluorescent vital probes, JC-1 (5,5',6,6' tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl-carbocyanine iodide) and oxidation sensitive dihydrorhodamine 123, and analyzed by flow cytometry to elucidate ethanol-induced time-wise alterations in the mitochondrial membrane potential (DeltaPsim) and the production of reactive oxygen species (ROS). Ethanol (50 and 200 mM) not only hyperpolarized DeltaPsim of both types of cells but also dose dependently increased ROS production at 24 h, although a 200-mM dose reduced the production until 3 h. These cell pathophysiological reactions suggest the depression of mitochondrial ATPase and mitochondrial respiratory chain. However, differences between these cells appeared after a 24-h exposure to 200 mM ethanol: the increase in ROS production was approximately twice as large for myocardial cells as for non-muscle cells; and the side-scatter parameter of light scattering significantly increased for myocardial cells, but not for non-muscle cells. All these myocyte-specific alterations indicate an increase in the mitochondrial fraction in a cell. This reaction might be a countermeasure against ethanol induced dysfunction of mitochondrial respiration that is needed to meet the energy requirements of spontaneous myocardial contractions. PMID- 16474960 TI - Study on the mechanism of photosensitive dermatitis caused by ketoprofen in the guinea pig. AB - To investigate the mechanism on photosensitive dermatitis caused by ketoprofen (KP) in humans, the following experiments were performed by topical application on guinea pigs. The phototoxicity study involving treatment with 10% solution of KP, its enantiomers (R-KP and S-KP), loxoprofen, and flurbiprofen revealed no phototoxic reactions. In the photoallergenicity study, KP and its enantiomers (0.5-2% solution) induced skin reaction at all dosages; however, loxoprofen and flurbiprofen (1-5% solution) did not induce such a photoallergenic reaction. These results suggest that the chemical structure of the benzophenone chromophore in KP would be one of the important factors for induction of the photoallergy since both loxoprofen and flurbiprofen do not possess this structure and hence lack photoallergenic potential. Furthermore, to assess time profiles of KP concentration in the skin and plasma, guinea pigs received a repeated topical application of R-KP and S-KP at a dosage of 40 mg/kg over a period of 3 days. Plasma KP concentrations were extremely low as compared to skin KP concentrations and were not detected at 72 h after the final dosing. At 24 h after the final dosing, KP concentrations in the skin with R-KP and S-KP treatment were 187.4 and 254.7 microg/g, respectively, and their half-lives were 80.5 and 84.4 h, respectively. KP concentrations at 336 h after final dosing were 11.3 microg/g for R-KP and 15.7 microg/g for S-KP treatment. The acylglycerol-combined KP concentrations at 336 h were 2% or less as compared to KP concentrations with R KP and S-KP treatment. There were no differences in KP concentrations in the skin between R-KP and S-KP and in combined KP concentrations between the enantiomers. The present study indicates that photosensitive dermatitis after topical application of KP in humans, caused by photoallergenicity and not phototoxicity, can be reproduced in the animal testing, and suggests that the skin reaction may be caused by the long period of retention of KP in the skin. PMID- 16474962 TI - Revisiting the population toxicokinetics of tetrachloroethylene. PMID- 16474961 TI - Sequencing and phylogenetic analysis of neurotoxin gene from an environmental isolate of Clostridium sp.: comparison with other clostridial neurotoxins. AB - A Clostridium sp. isolated from intestine of decaying fish exhibited 99% sequence identity with C. tetani at 16S rRNA level. It produced a neurotoxin that was neutralized by botulinum antitoxin (A+B+E) as well as tetanus antitoxin. The gene fragments for light chain, C-terminal and N-terminal regions of the heavy chain of the toxin were amplified using three reported primer sets for tetanus neurotoxin (TeNT). The neurotoxin gene fragments were cloned in Escherichia coli and sequenced. The sequences obtained exhibited approximately 98, 99 and 98% sequence identity with reported gene sequences of TeNT/LC, TeNT/HC and TeNT/HN, respectively. The phylogenetic interrelationship between the neurotoxin gene of Clostridium sp. with previously reported gene sequences of Clostridium botulinum A to G and C. tetani was examined by analysis of differences in the nucleotide sequences. Six amino acids were substituted at four different positions in the light chain of neurotoxin from the isolate when compared with the reported closest sequence of TeNT. Of these, four were located in the beta15 motif at a solvent inaccessible, buried region of the protein molecule. One of these substitutions were on the solvent accessible surface residue of alpha1 motif, previously shown to have strong sequence conservation. A substitution of two amino acids observed in N-terminal region of heavy chain were buried residues, located in the beta21 and beta37 motifs showing variability in other related sequences. The C-terminal region responsible for binding to receptor was conserved, showing no changes in the amino acid sequence. PMID- 16474963 TI - Isolation and biological characterization of neurotoxic compounds from the sea anemone Lebrunia danae (Duchassaing and Michelotti, 1860). AB - This paper describes two neurotoxic proteins obtained from the Caribbean sea anemone Lebrunia danae. To assess the neurotoxic activity of the venom of L. danae, several bioassays were carried out, and to evaluate the effect of the toxin, Median Lethal Doses (LD(50)) were determined in vivo using sea crabs (Ocypode quadrata) and Artemia salina nauplii with the crude extract of the proportion of 2.82 mg/m. The proteins with neurotoxic effects were isolated using low-pressure liquid chromatography. The fractions containing the neurotoxic activity were analyzed by SDS-PAGE and showed protein bands with an apparent molecular weight of 62.50 kDa (LdNt1) and 58 kDa (LdNt2). To demonstrate that these proteins were indeed responsible for the neurotoxic activity observed, we injected a small fraction of the purified protein into the third walking leg of a crab and observed the typical convulsions, paralysis and death provoked by neurotoxins. Hemolytic activity was also tested for 0.238 mg of crude extract; the hemolytic value was 39.5, 49.6 and 50.1% for cow, sheep and pig erythrocytes, respectively. PMID- 16474964 TI - New proposed responsibilities of a radiologist in the emergency room: what you need to know and must, shall, should do. PMID- 16474966 TI - Increased risk of obesity associated with the variant allele of the PPARGC1A Gly482Ser polymorphism in physically inactive elderly men. AB - AIMS/HYPOTHESIS: The variant allele of the Gly482Ser polymorphism in peroxisome proliferator-activated receptor-gamma co-activator-1alpha (PPARGC1A or PGC1alpha), a critical determinant of skeletal muscle metabolism, has been associated with obesity and type 2 diabetes. Previous studies indicate that these risks depend on sex and environmental triggers such as age. The aim of the present study was to investigate the possible interactions between genotype and age and physical activity on risk of obesity. METHODS: We genotyped PPARGC1A Gly482Ser, in a population-based study comprising 899 women and 902 men aged between 30 and 75 years in Vara, Sweden. RESULTS: Genotyping revealed that 56% of the males and 57% of the females carried the PPARGC1A 482Ser variant allele. Elderly males (>or=50 years) carrying 482Ser had an increased risk of obesity compared with subjects who were homozygous for the wild-type allele (odds ratio [OR]=1.99, 95% CI 1.14-3.47, p=0.015). The risk was restricted to males with a low leisure-time physical activity level, and was significantly weaker (OR=0.44, 95% CI 0.22-0.87, p=0.018) for the homozygous 482Gly carriers among this subgroup. No association with obesity was found in elderly males with a high level of physical activity, in younger males, or in females of any age group or level of physical activity. CONCLUSIONS/INTERPRETATION: Our findings confirm that sex and age should be considered when investigating the influence of the PPARGC1A Gly482Ser polymorphism on metabolic disease. The risk of obesity associated with 482Ser is evident only in physically inactive elderly male subjects. Whenever possible, the level of physical activity should be addressed in future studies on disease risk associated with PPARGC1A Gly482Ser. PMID- 16474967 TI - The reasons of general practitioners for promoting colorectal cancer mass screening in France. AB - Implementation of colorectal cancer (CRC) mass screening is a public health priority. Obtaining the involvement of general practitioners (GPs) is crucial for ensuring participation by the general public. This study elicited GPs' reasons for promoting CRC screening. The method used was discrete choice modeling. Questionnaires with a set of pairs of hypothetical screening scenarios were sent to 700 GPs. Factors influencing GPs' screening practice were found to be: the effectiveness of the screening program, the proportion of false negatives and false positives, and their remuneration. Hence improving screening test sensitivity and setting up specific remuneration for conducting screening would help to increase GPs' participation in CRC screening in France. PMID- 16474969 TI - Feeding responses to selected alkaloids by gypsy moth larvae, Lymantria dispar (L.). AB - Deterrent compounds are important in influencing the food selection of many phytophagous insects. Plants containing deterrents, such as alkaloids, are generally unfavored and typically avoided by many polyphagous lepidopteran species, including the gypsy moth Lymantria dispar (L.) (Lepidoptera: Lymantriidae). We tested the deterrent effects of eight alkaloids using two choice feeding bioassays. Each alkaloid was applied at biologically relevant concentrations to glass fiber disks and leaf disks from red oak trees (Quercus rubra) (L.), a plant species highly favored by these larvae. All eight alkaloids tested on glass fiber disks were deterrent to varying degrees. When these alkaloids were applied to leaf disks, only seven were still deterrent. Of these seven, five were less deterrent on leaf disks compared with glass fiber disks, indicating that their potency was dramatically reduced when they were applied to leaf disks. The reduction in deterrency may be attributed to the phagostimulatory effect of red oak leaves in suppressing the negative deterrent effect of these alkaloids, suggesting that individual alkaloids may confer context-dependent deterrent effects in plants in which they occur. This study provides novel insights into the feeding behavioral responses of insect larvae, such as L. dispar, to selected deterrent alkaloids when applied to natural vs artificial substrates and has the potential to suggest deterrent alkaloids as possible candidates for agricultural use. PMID- 16474968 TI - Economic impact of simplified de Gramont regimen in first-line therapy in metastatic colorectal cancer. AB - The cost of chemotherapy has dramatically increased in advanced colorectal cancer patients, and the schedule of fluorouracil administration appears to be a determining factor. This retrospective study compared direct medical costs related to two different de Gramont schedules (standard vs. simplified) given in first-line chemotherapy with oxaliplatin or irinotecan. This cost-minimization analysis was performed from the French Health System perspective. Consecutive unselected patients treated in first-line therapy by LV5FU2 de Gramont with oxaliplatin (Folfox regimen) or with irinotecan (Folfiri regimen) were enrolled. Hospital and outpatient resources related to chemotherapy and adverse events were collected from 1999 to 2004 in 87 patients. Overall cost was reduced in the simplified regimen. The major factor which explained cost saving was the lower need for admissions for chemotherapy. Amount of cost saving depended on the method for assessing hospital stay. In patients treated by the Folfox regimen the per diem and DRG methods found cost savings of Euro 1,997 and Euro 5,982 according to studied schedules; in patients treated by Folfiri regimen cost savings of Euro 4,773 and Euro 7,274 were observed, respectively. In addition, travel costs were also reduced by simplified regimens. The robustness of our results was showed by one-way sensitivity analyses. These findings demonstrate that the simplified de Gramont schedule reduces costs of current first-line chemotherapy in advanced colorectal cancer. Interestingly, our study showed several differences in costs between two costing approaches of hospital stay: average per diem and DRG costs. These results suggested that standard regimen may be considered a profitable strategy from the hospital perspective. The opposition between health system perspective and hospital perspective is worth examining and may affect daily practices. In conclusion, our study shows that the simplified de Gramont schedule in combination with oxaliplatin or irinotecan is an attractive option from the French Health System perspective. This safe and less costly regimen must compared to alternative options such as oral fluoropyrimidines. PMID- 16474970 TI - Acute pyomyositis of the pelvis: the spectrum of clinical presentations and MR findings. AB - BACKGROUND: Acute pelvic pyomyositis is uncommon in non-tropical areas. OBJECTIVE: To summarize the clinical and MR findings in children with acute pelvic pyomyositis. MATERIALS AND METHODS: We retrospectively identified 20 children (mean age 9.4 years) who were evaluated by MR and diagnosed with acute pelvis pyomyositis during the time period between January 2002 and June 2005. We reviewed clinical, laboratory, and imaging findings. RESULTS: Fifteen of the 20 children had secondary pyomyositis associated with osteomyelitis (n=13), septic hip (n=4) or sacroiliitis (n=4); all were previously healthy except for one child with leukemia. Seven of the children with secondary pyomyositis underwent bone scintigraphy; three (43%) did not show pelvic abnormalities. Staphylococcus aureus was cultured in 13 of the 15 (87%) children. Five of the 20 children had primary pyomyositis. Three had underlying disease and two others were engaged in vigorous physical activity. Bone scintigraphies (n=2) were negative. Cultures were positive for S. aureus in three of the five (60%) children. CONCLUSION: Septic hip should be the first diagnostic consideration in children with fever and acute hip pain. Pyomyositis should be considered if arthrocentesis is negative or there is clinical suspicion of infection outside the hip joint. MR is the preferred imaging modality for evaluating foci of pyomyositis, muscle abscesses, and additional foci of infection within the pelvis. PMID- 16474972 TI - Bilateral parotid gland tuberculosis. AB - Tuberculosis is caused by a mycobacterium and is one of the most serious diseases in the world. Pulmonary involvement is the most common form of this condition. Primary tuberculosis of the parotid gland is rare. We report such a case in a 34 year-old man who presented with a painless preauricular mass on each side of his head. The clinical presentation, diagnosis, pathological findings and treatment are described. PMID- 16474971 TI - Mining and characterizing microsatellites from citrus ESTs. AB - Freely available computer programs were arranged in a pipeline to extract microsatellites from public citrus EST sequences, retrieved from the NCBI. In total, 3,278 bi- to hexa-type SSR-containing sequences were identified from 56,199 citrus ESTs. On an average, one SSR was found per 5.2 kb of EST sequence, with the tri-nucleotide motifs as the most abundant. Primer sequences flanking SSR motifs were successfully identified from 2,295 citrus ESTs. Among those, a subset (100 pairs) were synthesized and tested to determine polymorphism and heterozygosity between/within two genera, sweet orange (C. sinensis) and Poncirus (P. trifoliata), which are the parents of the citrus core mapping population selected for an international citrus genomics effort. Eighty-seven pairs of primers gave PCR amplification to the anticipated SSRs, of which 52 and 35 appear to be homozygous and heterozygous, respectively, in sweet orange, and 67 and 20, respectively, in Poncirus. By pairing the loci between the two intergeneric species, it was found that 40 are heterozygous in at least one species with two alleles (9), three alleles (28), or four alleles (3), and the remaining 47 are homozygous in both species with either one allele (31) or two alleles (16). These EST-derived SSRs can be a resource used for understanding of the citrus SSR distribution and frequency, and development of citrus EST-SSR genetic and physical maps. These SSR primer sequences are available upon request. PMID- 16474973 TI - Oncocytic carcinoma of the maxillary sinus: a rare neoplasm. AB - Oncocytic neoplasms are tumors composed of oncocytes (i.e., epithelial cells with a large cytoplasm that is rich in mitochondria). Most cases are benign and originate from the major salivary glands, while the minor salivary glands are rarely involved. Occurrence of oncocytic carcinoma (or malignant oncocytoma) within the sinonasal tract is an unusual event. We report a rare case of maxillary sinus oncocytic carcinoma occurring in a 45-year-old male. Biopsy was consistent with an unspecified salivary gland neoplasm. The patient underwent total maxillectomy through a lateral rhinotomic approach; hard palate reconstruction with temporal myofascial flap was performed. Definitive histology was consistent with oncocytic carcinoma. Due to the local extension of the lesion, postoperative radiotherapy (60 Gy) was delivered. Three years after surgery, the patient is free from disease. A brief analysis of the literature was also accomplished in order to discuss treatment options and prognosis of this unusual neoplasm. PMID- 16474974 TI - Peroxisome proliferator-activated receptors-gamma activator, ciglitazone, inhibits human melanocyte growth through induction of apoptosis. AB - Peroxisome proliferator-activated receptors (PPARs) are members of the nuclear hormone receptor superfamily. All three PPAR subtypes, PPAR-alpha, PPAR beta/delta and PPAR-gamma are expressed in human melanocytes. In this study, we investigated the effects of PPAR-gamma activator on melanocyte growth, and apoptosis. The PPAR-gamma activators ciglitazone, troglitazone, and 15-deoxy prostaglandin J2 inhibited melanocyte growth in a dose-dependent manner. This inhibitory effect of ciglitazone seemed to occur through induction of apoptosis. Apoptosis was increased after ciglitazone treatment, which was observed by the TUNEL method and flow cytometry. We noted a decrease in extracellular signal regulated kinase protein expression under ciglitazone treatment. Western blot analysis revealed an apparent time-dependent reduction in Bcl-2 protein levels in ciglitazone-treated melanocytes. In terms of Bax expression, a difference was not found. The expression of caspase-3 proteins was increased time-dependently with ciglitazone treatment. These results indicate that melanocyte growth and apoptosis may be modulated through PPAR-gamma and that ciglitazone, a PPAR-gamma activator, inhibits growth of human melanocytes by inducing apoptosis. PMID- 16474975 TI - A novel chromosomal abnormality involving 2q37 in myeloid malignancies. PMID- 16474976 TI - Aplasia of the dorsal pancreas and choledochal cyst. AB - The authors describe a rare case of choledochal cyst and aplasia of the dorsal pancreas complicated with chronic pancreatitis. A 9-year-old boy presented with obstructive jaundice. After biliary drainage using pericutaneous transhepatic gallbladder drainage (PTGBD) technique, the patient underwent choledochal cyst excision with Roux-en-Y hepaticojejunostomy. The association of choledochal cyst and aplasia of the dorsal pancreas with chronic pancreatitis has never been reported previously. PMID- 16474978 TI - Molecular evolution and functionally important structures of molluscan Dermatopontin: implications for the origins of molluscan shell matrix proteins. AB - A major shell matrix protein originally obtained from a freshwater snail is a molluscan homologue of Dermatopontins, a group of Metazoan proteins also called TRAMP (tyrosine-rich acidic matrix protein). We sequenced and identified 14 molluscan homologues of Dermatopontin from eight snail species belonging to the order Basommatophora and Stylommatophora. The bassommatophoran Dermatopontins fell into three types, one is suggested to be a shell matrix protein and the others are proteins having more general functions based on gene expression analyses. N-glycosylation is inferred to be important for the function involved in shell calcification, because potential N-glycosylation sites were found exclusively in the Dermatopontins considered as shell matrix proteins. The stylommatophoran Dermatopontins fell into two types, also suggested to comprise a shell matrix protein and a protein having a more general function. Phylogenetic analyses using maximum likelihood and Bayesian methods revealed that gene duplication events occurred independently in both basommatophoran and stylommatophoran lineages. These results suggest that the dermatopontin genes were co-opted for molluscan calcification at least twice independently after the divergence of basommatophoran and stylommatophoran lineages, or more recently than we have expected. PMID- 16474980 TI - Phylogenetic and biochemical studies reveal a potential evolutionary origin of small heat shock proteins of animals from bacterial class A. AB - Small heat shock proteins (sHSPs), as one subclass of molecular chaperones, are important for cells to protect proteins under stress conditions. Unlike the large HSPs (represented by Hsp60 and Hsp70), sHSPs are highly divergent in both primary sequences and oligomeric status, with their evolutionary relationships being unresolved. Here the phylogenetic analysis of a representative 51 sHSPs (covering the six subfamilies: bacterial class A, bacterial class B, archae, fungi, plant, and animal) reveals a close relationship between bacterial class A and animal sHSPs which form an outgroup. Accumulating data indicate that the oligomers from bacterial class A and animal sHSPs appear to exhibit polydispersity, while those from the rest exhibit monodispersity. Together, the close evolutionary relationship and the similarity in oligomeric polydispersity between bacterial class A and animal sHSPs not only suggest a potential evolutionary origin of the latter from the former, but also imply that their oligomeric polydispersity is somehow a property determined by their primary sequences. PMID- 16474979 TI - A phylogenetic and structural analysis of truncated hemoglobins. AB - Truncated hemoglobins (trHbs) are heme proteins found in bacteria, plants, and unicellular eukaryotes. They are distantly related to vertebrate hemoglobins and are typically shorter than these by 20-40 residues. The multiple amino acid deletions, insertions, and replacements result in distinctive alterations of the canonical globin fold and a wide range of chemical properties. An early phylogenetic analysis categorized trHbs into three groups, I (trHbN), II (trHbO), and III (trHbP). Here, we revisit this analysis with 111 trHbs. We find that trHbs are orthologous within each group and paralogous across the groups. Group I globins form the most disparate set and separate into two divergent subgroups. Group II is comparatively homogeneous, whereas Group III displays the highest level of overall conservation. In Group I and Group II globins, for which some ligand binding and structural data are available, an improved description of probable protein-ligand interactions is achieved. Other conservation trends are either confirmed (essential glycines in loops), refined (lining of ligand access tunnel), or newly identified (helix start signal). The Group III globins, so far uncharacterized, exhibit recognizable heme cavity residues while lacking some of the residues thought to be important to the trHb fold. An analysis of the phylogenetic trees of each group provides a plausible scenario for the emergence of trHbs, by which the Group II trHb gene was the original gene, and the Group I trHb and Group III trHb genes were obtained via duplication and transfer events. PMID- 16474981 TI - The complete mitochondrial genome of the entomopathogenic nematode Steinernema carpocapsae: insights into nematode mitochondrial DNA evolution and phylogeny. AB - We determined the complete sequence of the mitochondrial DNA of the entomopathogenic nematode Steinernema carpocapsae and analyzed its structure and composition as well as the secondary structures predicted for its tRNAs and rRNAs. Almost the complete genome has been amplified in one fragment with long PCR and sequenced using a shotgun strategy. The 13,925-bp genome contains genes for 2 rRNAs, 22 tRNAs, and 12 proteins and lacks an ORF encoding ATPase subunit 8. Four initiation codons were inferred, TTT, TTA, ATA, and ATT, most of the genes ended with TAA or TAG, and only two had a T as an incomplete stop codon. All predicted tRNAs showed the nonconventional secondary structure typical of Secernentea. Although we were able to fold the sequences of trnN, trnD, and trnC into more conventional cloverleaf structures after adding adjacent nucleotides, northern blot experiments showed that the nonstandard tRNAs are actually expressed. Phylogenetic and comparative analyses showed that the mitochondrial genome of S. carpocapsae is more closely related to the genomes of A. suum and C. elegans than to that of Strongyloides stercoralis. This finding does not support the phylogeny based on nuclear small subunit ribosomal DNA sequences previously published. This discrepancy may result from differential reproductive strategies and/or differential selective pressure acting on nuclear and mitochondrial genes. The distinctive characteristics observed among mitochondrial genomes of Secernentea may have arisen to counteract the deleterious effects of Muller's ratchet, which is probably enhanced by the reproductive strategies and selective pressures referred to above. PMID- 16474982 TI - Asymmetrical evolution of cytochrome bd subunits. AB - Functionally linked genes generally evolve at similar rates and the knowledge of this particular feature of genomic evolution has been used as the basis for the phylogenetic profiling method. We illustrate here an exception to this rule in the evolution of the cytochrome bd complex. This is a two-component oxidase complex, with the subunits I and II known to be widely present in bacteria. The subunits within the cytochrome bd complex are under the same evolutionary pressure and most likely behave in the same evolutionary manner. However, the sequence similarity of genes encoding subunit II varies considerably across species. Genes encoding subunit II evolve 1.2 times faster on most of the branches of their phylogeny than subunit I genes. Furthermore, the genes encoding subunit II in Oceanobacillus iheyensis, Bacillus halodurans, and Staphylococcus species do not have detectable homologues within E. coli due to their large divergence. Together, the two subunits of cytochrome bd reveal an interesting example of an asymmetric pattern of evolutionary change. PMID- 16474984 TI - Adaptive evolution of multicolored fluorescent proteins in reef-building corals. AB - Here we investigate the evolutionary scenarios that led to the appearance of fluorescent color diversity in reef-building corals. We show that the mutations that have been responsible for the generation of new cyan and red phenotypes from the ancestral green were fixed with the help of positive natural selection. This fact strongly suggests that the color diversity is a product of adaptive evolution. An unexpected finding was a set of residues arranged as an intermolecular binding interface, which was also identified as a target of positive selection but is nevertheless not related to color diversification. We hypothesize that multicolored fluorescent proteins evolved as part of a mechanism regulating the relationships between the coral and its algal endosymbionts (zooxanthellae). We envision that the effect of the proteins' fluorescence on algal physiology may be achieved not only through photosynthesis modulation, but also through regulatory photosensors analogous to phytochromes and cryptochromes of higher plants. Such a regulation would require relatively subtle, but spectrally precise, modifications of the light field. Evolution of such a mechanism would explain both the adaptive diversification of colors and the coevolutionary chase at the putative algae-protein binding interface in coral fluorescent proteins. PMID- 16474983 TI - Long repeats in a huge genome: microsatellite loci in the grasshopper Chorthippus biguttulus. AB - It is commonly believed that both the average length and the frequency of microsatellites correlate with genome size. We have estimated the frequency and the average length for 69 perfect dinucleotide microsatellites in an insect with an exceptionally large genome: Chorthippus biguttulus (Orthoptera, Acrididae). Dinucleotide microsatellites are not more frequent in C. biguttulus, but repeat arrays are 1.4 to 2 times longer than in other insect species. The average repeat number in C. biguttulus lies in the range of higher vertebrates. Natural populations are highly variable. At least 30 alleles per locus were found and the expected heterozygosity is above 0.95 at all three loci studied. In contrast, the observed heterozygosity is much lower (< or = 0.51), which could be caused by long null alleles. PMID- 16474985 TI - Substitution rate heterogeneity and the male mutation bias. AB - Germline mutation rates have been found to be higher in males than in females in many organisms, a likely consequence of cell division being more frequent in spermatogenesis than in oogenesis. If the majority of mutations are due to DNA replication error, the male-to-female mutation rate ratio (alpha(m)) is expected to be similar to the ratio of the number of germ line cell divisions in males and females (c), an assumption that can be tested with proper estimates of alpha(m) and c. Alpha(m) is usually estimated by comparing substitution rates in putatively neutral sequences on the sex chromosomes. However, substantial regional variation in substitution rates across chromosomes may bias estimates of alpha(m) based on the substitution rates of short sequences. To investigate regional substitution rate variation, we estimated sequence divergence in 16 gametologous introns located on the Z and W chromosomes of five bird species of the order Galliformes. Intron ends and potentially conserved blocks were excluded to reduce the effect of using sequences subject to negative selection. We found significant substitution rate variation within Z chromosome (G15 = 37.6, p = 0.0010) as well as within W chromosome introns (G15 = 44.0, p = 0.0001). This heterogeneity also affected the estimates of alpha(m), which varied significantly, from 1.53 to 3.51, among the introns (ANOVA: F(13,14) = 2.68, p = 0.04). Our results suggest the importance of using extensive data sets from several genomic regions to avoid the effects of regional mutation rate variation and to ensure accurate estimates of alpha(m). PMID- 16474986 TI - Molecular evolution and positive selection of the symbiotic gene NORK in Medicago truncatula. AB - Understanding the selective constraints of partner specificity in mutually beneficial symbiosis is a significant, yet largely unexplored, prospect of evolutionary biology. These selective constraints can be explored through the study of nucleotide polymorphism at loci controlling specificity. The membrane anchored receptor NORK (nodulation receptor kinase) of the legume Medicago truncatula controls early steps of root infection by two symbiotic microorganisms: nitrogen-fixing bacteria (rhizobia) and endomycorrhizal fungi (Glomales). We analyzed the diversity of the gene NORK by sequencing 4 kilobases in 28 inbred lines sampled from natural populations. We detected 33 polymorphic sites with only one nonsynonymous change. Analysis based on Tajima's D and Fay and Wu's H summary statistics revealed no departure from the neutral model. We analyzed divergence using sequences from the closely related species M. coerulea. The McDonald-Kreitman test indicated a significant excess of nonsynonymous changes contributing to this divergence. Furthermore, maximum-likelihood analysis of a molecular phylogeny of a few legume species indicated that a number of amino acid sites, likely located in the receptor domain of the protein, evolved under the regime of positive selection. Further research should focus on the rate and direction of molecular coevolution between microorganisms' signaling molecules and legumes' receptors. PMID- 16474988 TI - Anal electrical stimulation with long pulses increases anal sphincter pressure in conscious dogs. AB - PURPOSE: This study was designed to investigate the effects and mechanisms of anal electric stimulation with long pulses on anal sphincter pressure in conscious dogs. METHODS: The study was performed after enema in nine healthy female hound dogs and composed of four randomized sessions ("dose"-response, anal electric stimulation only, or with atropine or phentolamine). The anal sphincter pressure was measured by using manometry and quantified by using the area under the contractile curve (mmHg/sec). Anal electric stimulation was performed via a pair of ring electrodes attached to a manometric catheter. The stimulation parameters in all but dose-response sessions included a frequency of 20 ppm, pulse width of 200 ms, and amplitude of 3 mA. RESULTS: The anal sphincter pressure was 55.7 +/- 6 at baseline and increased by 37 percent to 76.4 +/- 6.5 during electric stimulation (P = 0.009). The increase of anal pressure during stimulation was positively correlated with the stimulation energy (r = 0.395; P < 0.01). The excitatory effect of electric stimulation was sustained for at least 20 minutes. Atropine did not alter anal pressure and did not abolish the excitatory effect of anal electric stimulation on the sphincter. Phentolamine reduced anal pressure from the baseline value of 50.5 +/- 4.7 to 33.1 +/- 5.4 (P = 0.019). The electric stimulation induced increase in anal pressure was dropped from 19 +/- 2.6 to 9.9 +/- 2.8 (P = 0.029) at the presence of phentolamine. CONCLUSIONS: Anal electric stimulation with long pulses increases anal sphincter pressure in an energy-dependent manner. The alpha-adrenergic but not the cholinergic pathway at least partially mediates the excitatory effect of anal electric stimulation. PMID- 16474987 TI - A "green" phosphoribulokinase in complex algae with red plastids: evidence for a single secondary endosymbiosis leading to haptophytes, cryptophytes, heterokonts, and dinoflagellates. AB - Phosphoribulokinase (PRK) is an essential enzyme of photosynthetic eukaryotes which is active in the plastid-located Calvin cycle and regenerates the substrate for ribulose-bisphosphate carboxylase/oxygenase (Rubisco). Rhodophytes and chlorophytes (red and green algae) recruited their nuclear-encoded PRK from the cyanobacterial ancestor of plastids. The plastids of these organisms can be traced back to a single primary endosymbiosis, whereas, for example, haptophytes, dinoflagellates, and euglenophytes obtained their "complex" plastids through secondary endosymbioses, comprising the engulfment of a unicellular red or green alga by a eukaryotic host cell. We have cloned eight new PRK sequences from complex algae as well as a rhodophyte in order to investigate their evolutionary origin. All available PRK sequences were used for phylogenetic analyses and the significance of alternative topologies was estimated by the approximately unbiased test. Our analyses led to several astonishing findings. First, the close relationship of PRK genes of haptophytes, heterokontophytes, cryptophytes, and dinophytes (complex red lineage) supports a monophyletic origin of their sequences and hence their plastids. Second, based on PRK genes the complex red lineage forms a highly supported assemblage together with chlorophytes and land plants, to the exclusion of the rhodophytes. This green affinity is in striking contrast to the expected red algal origin and our analyses suggest that the PRK gene was acquired once via lateral transfer from a green alga. Third, surprisingly the complex green lineages leading to Bigelowiella and Euglena probably also obtained their PRK genes via lateral gene transfers from a red alga and a complex alga with red plastids, respectively. PMID- 16474989 TI - Coexpression of VEGF-C and Cox-2 in human colorectal cancer and its association with lymph node metastasis. AB - PURPOSE: Several lines of experimental evidence indicated that over-expression of vascular endothelial growth factor-C and cyclooxygenase-2 genes promotes angiogenesis and lymphangiogenesis, both of which are essential for the growth and spreading of tumor cells. This study was designed to evaluate the coexpression of vascular endothelial growth factor-C and cyclooxygenase-2 in human colorectal carcinoma to determine their relationships and correlations with lymph node metastasis and prognosis. METHODS: Tissue samples of primary tumors and metastatic lymph nodes from 150 patients undergoing intentionally curative surgical resections for colorectal adenocarcinoma were immunohistochemically examined for vascular endothelial growth factor-C, cyclooxygenase-2, and CD34 expressions. Then, we analyzed their relationships and correlations with clinicopathologic findings and patients' survival time. RESULTS: The positivity rate of vascular endothelial growth factor-C and cyclooxygenase-2 in the primary tumor was 68 and 72.7 percent, respectively, and in the metastatic lymph nodes was 93.3 and 80 percent, respectively. A significant correlation was found between the expression scores of vascular endothelial growth factor-C and cyclooxygenase-2 (P < 0.0001), and both also were correlated to microvessels density and several clinicopathologic parameters, including primary tumor size, lymph node metastasis, lymphatic invasion, and TNM stage. Patients with vascular endothelial growth factor-C-positive and/or cyclooxygenase-2-positive tumors had a significant shorter survival time than those with negative tumors did. However, in a multivariate analysis, only cyclooxygenase-2 expression was recognized as an independent prognostic factor (P = 0.0412; relative risk ratio, 3.067; 95 percent confidence interval, 1.046-8.994). CONCLUSIONS: These data show that in human colorectal carcinoma, vascular endothelial growth factor-C and cyclooxygenase-2 are coexpressed and significantly associated with lymph node metastasis and prognosis. PMID- 16474990 TI - Behavioral problems and the effects of early intervention on eight-year-old children with learning disabilities. AB - OBJECTIVES: 1) To investigate the comorbidity of verbal and nonverbal learning disability subtypes with several domains of behavior problems among 8-year-old children. 2) To determine whether receipt of an early intervention modified the association between childhood behavior problems and learning disabilities (LD). METHODS: This is a secondary data analysis of the Infant Health and Development Program (IHDP), a randomized clinical trial of an early intervention provided between ages 0 and 3 involving 985 children born low birthweight and premature. The findings are based on a prospective follow-up of these children at 8 years of age. RESULTS: Compared to children without verbal LD (VLD), those with VLD were twice as likely to exhibit clinical levels of total behavior problems and 89% more likely to exhibit externalizing behavior problems. Analysis of specific subscales of behavior revealed significant associations with anxious/depressed and withdrawn behaviors, as well as an increased likelihood of attention problems among children with VLD. No significant association was found between nonverbal LD (NVLD) and any type of behavior problem. Furthermore, there was a significant interaction between VLD and the intervention, in which the odds of internalizing behavior problems were greater among children with VLD. No interaction effect of the intervention occurred for any type of behavior problem among children with NVLD. CONCLUSIONS: These findings provide evidence that distinct differences exist for different learning disability subtypes with regards to behavioral outcomes and the effects of early intervention services among 8-year-old children. PMID- 16474991 TI - Neuroprotective effect of a chuk-me-sun-dan on neurons from ischemic damage and neuronal cell toxicity. AB - Chukmesundan (CMSD), composed of the following 8 medicinal herbs including Panex ginseng C.A. MEYER, Atractylodes macrocephala KOID, Poria cocos WOLF, Pinellia ternata BREIT, Brassica alba BOISS, Aconitum carmichaeli DEBX, Cynanchum atratum BGE and Cuscuta chinensis LAM. CMSD is being used in Korea for the treatment of various symptoms accompanying hypertension and cerebrovascular disorders. This study was carried out to examine the effects of CMSD on cultured primary neuron cell, cell cytotoxicity and lipid peroxidation in Abeta-treated cells. Cell death was enhanced by addition of Abeta. Pretreatment of CMSD attenuated in cell killing induced by Abeta. The protective effect of the CMSD water extracts on Abeta-induced neuronal death was also observed by lactate dehydrogenase assay using cultured astrocyte cells. Abeta-induced cell death was protected by the water extract of CMSD in a dose-dependent manner, and 25-50 microg/ml was the most effective concentration. CMSD has been also shown to protect primary cultured neurons from N-methyl-D: -aspartate receptor-mediated glutamate toxicity. It was in vivo evidenced that CMSD protects neurons against ischemia induced cell death. Moreover, oral administration of CMSD into mice prevented ischemia-induced learning disability and rescued hippocampal CA1 neurons from lethal ischemic damage. The neuroprotective action of exogenous CMSD was also confirmed by counting synapses in the hippocampal CA1 region. The presence of CMSD in neuron cultures rescued the neurons from nitrogen oxide (NO)-induced death. From these, it was suggested that CMSD may exert its neuroprotective effect by reducing the NO-mediated formation of free radicals or antagonizing their toxicity. PMID- 16474992 TI - Nitric oxide production in striatum and pallidum of cirrhotic rats. AB - Ammonium and manganese are neurotoxic agents related to brain metabolic disturbances observed after prolonged liver damage. The aim of this study was to assess the production of nitric oxide (NO) in the brain of cirrhotic rats exposed to manganese. We induced cirrhosis by bile duct ligation for 4 weeks in rats. From brain, striatum and globus pallidus were dissected out, and NO synthase activity and the content of nitrites plus nitrates (NOx) were determined. In pallidum we found a diminished constitutive NO synthase activity from cirrhotic rats, independently of manganese exposure. This result was confirmed by low levels of NOx in the same brain area (P<0.05, two-way ANOVA). This finding was not related to protein expression of NO synthase since no differences were observed in immunoblot signals between cirrhotic and sham-operated animals. Results from present study suggest that the production of NO is reduced in basal ganglia during cirrhosis. PMID- 16474993 TI - Neuroprotective effect of A20 on TNF-induced postischemic apoptosis. AB - Focal cerebral ischemia causes apoptosis in neural cells during the postischemia period. TNF is critically involved in such neuronal apoptosis mediated by caspase pathways. A20 can inhibit TNF-induced apoptosis in many cell types. However, little work has been carried out in central nervous system. In the present study, gene transfer of A20 resulted in reduction of infarct volume and improvement of neurological deficit in ischemia rats. Results of flow cytometry, TUNEL and DNA fragmentation assay all indicated A20 could inhibit TNF-induced apoptosis both in primary rat hippocampal neurons and SH-SY5Y cells. Moreover, we found A20 targeted the TNF apoptotic pathway by inhibiting proteolytic cleavage of caspase 8 and 3 in SH-SY5Y cells. These data demonstrated A20 could effectively protect neurons from postischemic apoptosis and may function partly on death receptor caspase pathway. Gene transfer of A20 may be a promising approach to gene therapy for cerebral ischemia in the future. PMID- 16474994 TI - Increased response to high KCl-induced elevation in the intracellular-Ca(2+) concentration in differentiated NG108-15 cell and the inhibitory effect of the L type Ca(2+) channel blocker, calciseptine. AB - Characteristics of the increasing effect for the concentration of intracellular calcium ions ([Ca(2+)](i)) by high-KCl application were investigated in the neuroblastomaxglioma hybrid NG108-15 cell line (NG108-15 cells). The present study confirmed that the increasing effect of [Ca(2+)](i) by high-KCl application in single NG108-15 cells, differentiated with dibutyryl cAMP (Bt(2)cAMP), was significantly enhanced, compared to undifferentiated cells. The following observations were made at first: (1) The response to high-KCl application, in both undifferentiated and differentiated cells, was significantly inhibited by calciseptine (CaS), an L-type Ca(2+) channel blocker, but not by N-, P- and R type Ca(2+) channel blockers. The IC(50) values for CaS in both undifferentiated and differentiated cell was almost identical. (2) The inhibitory effect of CaS was irreversible. (3) The increasing effect for [Ca(2+)](i) by high-KCl application was completely dependent on the presence of extracellular calcium ions. (4) The increased [Ca(2+)](i) by high-KCl application under a plateau concentration was quickly decreased to basal levels when the high-KCl solution was exchanged for a high-KCl solution containing EGTA (without CaCl(2)). Together, these results suggest that the enhancement of the response effect of [Ca(2+)](i) by high-KCl application in differentiated single NG108-15 cells was mainly due to the quantitative increase of L-type voltage-sensitive calcium channels (VSCCs), which were irreversibly inhibited by CaS. PMID- 16474995 TI - Differential effects of polyunsaturated fatty acids on membrane capacitance and exocytosis in rat pheochromocytoma-12 cells. AB - The fusion of synaptic vesicles with the plasma membrane during exocytosis can be recorded by membrane capacitance measurements under voltage-clamp conditions. These measurements enable high time-resolution quantitation of exocytosis. The present study was carried out using the above technique to elucidate the effects of various polyunsaturated fatty acids on exocytosis in a neuroendocrine cell, the rat pheochromocytoma-12 (PC12) cell. External application of eicosapentaenoic acid and arachidonic acid resulted in an increase in capacitance of PC12 cells, indicating fusion of secretory vesicles with cell membranes and exocytosis. In contrast, docosahexaenoic acid, linoleic acid, oleic acid, and vehicle control had no significant effect on capacitance. The above findings show differential effects of polyunsaturated fatty acids on exocytosis in PC12 cells. It is postulated that besides arachidonic acid, eicosapentaenoic acid could also play an important role in exocytosis and neurotransmitter release, in neurons and hormone-secreting cells. PMID- 16474996 TI - Nitric oxide produced during sublethal ischemia is crucial for the preconditioning-induced down-regulation of glutamate transporter GLT-1 in neuron/astrocyte co-cultures. AB - In the brain, prior sublethal ischemia (preconditioning, PC) produces tolerance of neurons to subsequent lethal ischemia. This study aims at elucidating whether and how nitric oxide (NO) produced during PC is involved in the PC-induced ischemic tolerance of neurons in neuron/astrocyte co-cultures. The rise in the extracellular concentration of glutamate during ischemia caused by the reversed uptake of glutamate (Glu) by the astrocytic Glu transporter GLT-1 was markedly suppressed by the prior PC treatment, but the suppression was reversed by treatment with an inhibitor of nitric oxide synthase (NOS) during PC. Immunocytochemical and Western blot analyses demonstrated that the expression of GLT-1 was down-regulated after the PC insult, and this down-regulation was also antagonized by treatment with NOS inhibitors during PC. Here we show that nNOS derived NO produced during PC was crucial for the down-regulation of astrocytic GLT-1, and this down-regulation coincided with an increased survival rate of neurons. PMID- 16474997 TI - Effects of magnesium sulfate administration during hypoxia on CaM kinase IV and protein tyrosine kinase activities in the cerebral cortex of newborn piglets. AB - The present study tested the hypothesis that magnesium sulfate administration prior to hypoxia prevents hypoxia-induced increase in Ca(2+)/Calmodulin-dependent kinase (CaM Kinase) IV and Protein Tyrosine Kinase (PTK ) activities. Animals were randomly divided into normoxic (Nx), hypoxic (Hx) and magnesium-pretreated hypoxic (Mg(2+)-Hx) groups. Cerebral hypoxia was confirmed biochemically by measuring ATP and phosphocreatine (PCr) levels. CaM Kinase IV and PTK activities were determined in Nx, Hx and Mg(2+)-Hx newborn piglets. There was a significant difference between CaM kinase IV activity (pmoles/mg protein/min) in Nx (270 +/- 49), Mg(2+)-Hx (317 +/- 82) and Hx (574 +/- 41, P < 0.05 vs. Nx and Mg(2+)-Hx) groups. Similarly, there was a significant difference between Protein Tyrosine Kinase activity (pmoles/mg protein/h) in normoxic (378 +/- 68), Mg(2+)-Hx (455 +/ 67) and Hx (922 +/- 66, P < 0.05 vs. Nx and Mg(2+)-Hx ) groups. We conclude that magnesium sulfate administration prior to hypoxia prevents hypoxia-induced increase in CaM Kinase IV and Protein Tyrosine Kinase activities. We propose that by blocking the NMDA receptor ion-channel mediated Ca(2+)-flux, magnesium sulfate administration inhibits the Ca(2+)/calmodulin-dependent activation of CaMKIV and prevents the generation of nitric oxide free radicals and the subsequent increase in PTK activity. As a result, phosphorylation of CREB and Bcl-2 family of proteins is prevented leading to prevention of programmed cell death. PMID- 16474998 TI - Effects of magnesium sulfate administration during hypoxia on Ca(2+) influx and IP(3) receptor modification in cerebral cortical neuronal nuclei of newborn piglets. AB - Magnesium is a non-competitive antagonist of the NMDA receptor. Hypoxic insults to the brain are associated with a significant increase in the intranuclear Ca(2+) due to altered nuclear membrane Ca(2+) influx mechanisms including hypoxia induced modifications of nuclear membrane IP(3) receptors. In this study we have examined the effects of magnesium sulfate administration to newborn piglets subjected to normoxia and severe hypoxia. The animals were randomly divided into normoxic (n=4), hypoxic (n=4) and magnesium sulfate treated hypoxic (n=4) groups. Hypoxia was confirmed biochemically by measuring ATP and phosphocreatine (PCr) levels in the brain tissue. Intranuclear Ca(2+) influx was assessed by measuring (45)Ca(2+) uptake. Results show a significant (P<0.05) decrease in ATP and PCr levels in hypoxic group in comparison with normoxia. On the other hand magnesium treated hypoxic group showed a significantly (P<0.05) higher ATP and PCr in comparison with the hypoxic group. Intranuclear Ca(2+) was significantly (P<0.05) higher in the hypoxic group in comparison with both normoxic and magnesium treated hypoxic groups. In addition results show that magnesium prevented hypoxia induced modification of the IP(3) receptor. Magnesium treatment significantly reduced the hypoxia-induced increase in the number of receptors (reduced B(max) - P<0.05-treated hypoxia vs. hypoxia and normoxia), and reversed the receptor affinity (reduced dissociation coefficient-K(d)--P<0.05-treated hypoxia vs. normoxia). The results demonstrate that the administration of magnesium sulfate prior to hypoxia prevents the hypoxia-induced increase in intranuclear Ca(2+) and IP(3) receptor modifications. We conclude that Mg(2+ )administration prevents hypoxia-induced modification of neuronal nuclear membrane function that leads to intranuclear Ca(2+)-dependent transcription of apoptotic proteins leading to hypoxic neuronal programmed cell death. PMID- 16474999 TI - Carbon disulfide-induced changes in cytoskeleton protein content of rat cerebral cortex. AB - To investigate the mechanism of carbon disulfide-induced neuropathy, male wistar rats were administrated by gavage at dosage of 300 or 500 mg/kg carbon disulfide, five times per week for 12 weeks. By the end of the exposure, the animals produced a slight or moderate level of neurological deficits, respectively. Cerebrums of carbon disulfide-intoxicated rats and their age-matched controls were Triton-extracted and centrifuged at a high speed (100,000 x g) to yield a pellet fraction of NF polymer and a corresponding supernatant fraction, which presumably contained mobile monomer. Then, the contents of six cytoskeletal protein (NF-L, NF-M, NF-H, alpha-tubulin, beta-tubulin, and beta-actin) in both fractions were determined by immunoblotting. Results showed that the contents of the three neurofilament subunits in the pellet and the supernatant fraction decreased significantly regardless of dose levels (P<0.01). As for microtubule proteins, in the pellet fraction of cerebrum, the levels of alpha-tubulin and beta-tubulin demonstrated some inconsistent changes. However, in the supernatant fractions, the content of alpha-tubulin and beta-tubulin increased significantly in both two dose groups (P<0.01). In comparison to neurofilament and tubulin proteins, the content of beta-actin changed less markedly, only the supernatant fraction of the high dose group displayed significant increase (P<0.01), but the others remained unaffected. These findings suggested that the changes of cytoskeleton protein contents in rat cerebrum were associated with the intoxication of carbon disulfide, which might be involved in the development of carbon disulfide neurotoxicity. PMID- 16475000 TI - Serotonin turnover in discrete regions of young rat brain after 24 h REM sleep deprivation. AB - The present study has attempted to elucidate the alteration of serotonin turnover after 24 h REM sleep deprivation in different regions in brain of young rat. Sleep deprivation was induced by the inverted flower pot technique. Results of this study show increased serotonin turnover after 24 h REM sleep deprivation in all the brain regions except in the hypothalamus. The decreased 5-HT ratio shows increased serotonin in the hypothalamus after 24 h sleep deprivation. This study indicates increased activity of serotonergic neurons in the hypothalamus after 24 h sleep deprivation. This also indicates that the hypothalamus plays a role in the immediate compensatory mechanism during 24 h REM sleep deprivation in young rats. PMID- 16475001 TI - Reactive macrophages increase oxidative stress and alpha-synuclein nitration during death of dopaminergic neuronal cells in co-culture: relevance to Parkinson's disease. AB - Parkinson's disease (PD) is characterized by progressive degeneration of dopaminergic neurons and a substantial decrease in the neurotransmitter dopamine in the nigro-striatal region of the brain. Increased markers of oxidative stress, activated microglias and elevated levels of pro-inflammatory cytokines have been identified in the brains of patients with PD. Although the precise mechanism of loss of neurons in PD remains unclear, these findings suggest that microglial activation may contribute directly to loss of dopaminergic neurons in PD patients. In the present study, we tested the hypothesis that activated microglia induces nitric oxide-dependent oxidative stress which subsequently causes death of dopaminergic neuronal cells in culture. We employed lipopolysaccharide (LPS) stimulated mouse macrophage cells (RAW 264.7) as a reactive microglial model and SH-SY5Y cells as a model for human dopaminergic neurons. LPS stimulation of macrophages led to increased production of nitric oxide in a time and dose dependent manner as well as subsequent generation of other reactive nitrogen species such as peroxynitrite anions. In co-culture conditions, reactive macrophages stimulated SH-SY5Y cell death characterized by increased peroxynitrite concentrations and nitration of alpha-synuclein within SH-SY5Y cells. Importantly 1,400 W, an inhibitor of the inducible nitric oxide synthase provided protection from cell death via decreasing the levels of nitrated alpha synuclein. These results suggest that reactive microglias could induce oxidative stress in dopaminergic neurons and such oxidative stress may finally lead to nitration of alpha-synuclein and death of dopaminergic neurons in PD. PMID- 16475002 TI - A novel aminopeptidase with highest preference for lysine. AB - Neuropeptides are formed from sedentary precursors to smaller, active peptides by processing enzymes cleaving at paired basic residues. The process generates peptide intermediates with additional Lys or Arg residues at their NH(2) and COOH termini; the N-terminal basic amino acids are later removed by specific aminopeptidases. We report here a novel lysine-specific aminopeptidase (KAP) of ubiquitous distribution. The enzyme was resolved from puromycin-sensitive aminopeptidase (PSA), aminopeptidase B (APB), and neuron-specific aminopeptidase (NAP). It was purified by FPLC after (NH(4))(2)SO(4) precipitation. The purified KAP had a K(m) of 333 microM with a V(max) of 0.7 nmol Lys ssNA/min/mg protein. N terminal basic amino acids, Lys in particular, were its favorable substrates. KAP was inhibited by chelating agents and by serine protease inhibitors. It was highly sensitive to aminopeptidase inhibitor bestatin, but insensitive to puromycin and amastatin, showing that KAP is distinct from PSA, NAP, and aminopeptidase A (APA). The 62,000-Da enzyme had a pH optimum at 7.5 and NaCl was its strongest activator. However, metals could not restore KAP's activity after it was dialyzed against EGTA. Our data indicated that rat KAP did not resemble any aminopeptidases as well as the microbial lysine aminopeptidases. PMID- 16475003 TI - Glucose metabolism in rat retinal pigment epithelium. AB - The retinal pigment epithelium (RPE) is the major transport pathway for exchange of metabolites and ions between choroidal blood supply and the neural retina. To gain insight into the mechanisms controlling glucose metabolism in RPE and its possible relationship to retinopathy, we studied the influence of different glucose concentrations on glycogen and lactate levels and CO(2) production in RPE from normal and streptozotocin-treated diabetic rats. Incubation of normal RPE in the absence of glucose caused a decrease in lactate production and glycogen content. In normal RPE, increasing glucose concentrations from 5.6 mM to 30 mM caused a four-fold increase in glucose accumulation and CO(2) yield, as well as reduction in lactate and glycogen production. In RPE from diabetic rats glucose accumulation did not increase in the presence of high glucose substrate, but it showed a four- and a seven-fold increase in CO(2) production through the mitochondrial and pentose phosphate pathways, respectively. We found high glycogen levels in RPE which can be used as an energy reserve for RPE itself and/or neural retina. Findings further show that the RPE possesses a high oxidative capacity. The large increase in glucose shunting to the pentose phosphate pathway in diabetic retina exposed to high glucose suggests a need for reducing capacity, consistent with increased oxidative stress. PMID- 16475004 TI - Salviae Miltiorrhizae BGE Radix increases rat striatal K(+)-stimulated dopamine release and activates the dopamine release with protection against hydrogen peroxide-induced injury in rat pheochromocytoma PC12 cells. AB - The present study investigated the effect of the medicinal plant Salviae miltiorrhizae radix (SMR) on dopaminergic neurotransmission in comparison with amphetamine. The effect of SM (0.1 g/ml) on K(+) (20 mM)-stimulated dopamine (DA) release from rat striatal slices was compared with amphetamine (10(-4) M). Amphetamine and SMR significantly increased K(+)-stimulated DA release (P<0.001) from rat striatal slices when compared with K(+)-stimulated alone. On the other hand, to examine whether in vitro SMR treatment induces DA release in PC12 cells, the role of protein kinases has been investigated in the induction of the SMR mediated events by using inhibitors of protein kinase C (PKC), mitogen activated protein kinase (MAP kinase) or protein kinase A (PKA). PKC inhibitors chelerythrine (50 and 100 nM), Ro31-8220 (100 nM) and the MAP kinase inhibitor, PD98059 (20 microM) inhibited the ability of SMR to elicit the SMR-stimulated DA release. The direct-acting PKC activator, 12-O-tetradecanoyl phorbol 13-acetate (TPA, 100 nM) mimicked the ability of SMR to elicit DA release. On the contrary, a selective PKA inhibitor, 50 microM Rp-8-Br-cAMP, blocked the development of SMR stimulated DA release. The results demonstrated that SMR may stimulate DA release and that SMR-induced increases in MAP kinase and PKC are important for induction of the enhancement in transporter-mediated DA release and PKA was also required for the enhancement in SMR-stimulated DA release. SMR treatment (0.1-10 microg/ml) to the hydrogen peroxide (H(2)O(2))-treated PC12 cells activated the enzyme activities such as catalase, superoxide dismutase and glutathione peroxidase, and decreased the malondialdehyde level, indicating that SMR has also protective effects against free radical-induced cell toxicity. Therefore, the mechanism by which SMR induces the enhancement in SMR-stimulated DA release is apparent. It remains to be determined whether the effect of SMR on DA function is important in its therapeutic use in the treatment of drug addiction. PMID- 16475005 TI - Six years after the commercial introduction of Bt maize in Spain: field evaluation, impact and future prospects. AB - We carried out a 6-year-field evaluation to assess potential hazards of growing Compa, a transgenic Bt maize variety based on the transformation event CG 00256 176. Two categories of hazards were investigated: the potential of the target corn borer Sesamia nonagrioides to evolve resistance to Bt maize and effects on non-target organisms. In order to address the first hazard, dispersal capacity of the corn borer was measured and our results indicated that larvae move to plants other than those onto which the female oviposited - even to plants in adjacent rows - in remarkable numbers and they do so mostly at a mature age, suggesting that mixing Bt and non-Bt seeds in the same field would not be a very useful deployment strategy to delay/prevent resistance. In addition, adults move among fields to mate and males may do so for up to 400 m. Three different aspects of potential non-target effects were investigated: sub-lethal effects on the target S. nonagrioides, effects on non-target maize pests, and effects on maize-dwelling predators. Larvae collected in Bt fields at later growth stages, in which event 176 Bt maize expresses Bt toxin at sub-lethal concentrations, had longer diapause and post-diapause development than larvae collected in non-Bt fields, a feature that might lead to a certain isolation between populations in both type of fields and accelerate Bt resistance evolution. Transgenic maize did not have a negative impact on non-target pests in the field; more aphids and leafhoppers but similar numbers of cutworms and wireworms were counted in Bt versus non-Bt fields; in any case differences in damage or yield were recorded. We observed no difference in the numbers of the most relevant predators in fields containing transgenic or no transgenic maize. PMID- 16475006 TI - Transgenic plants for insect pest control: a forward looking scientific perspective. AB - One of the first successes of plant biotechnology has been the creation and commercialisation of transgenic crops exhibiting resistance to major insect pests. First generation products encompassed plants with single insecticidal Bt genes with resistance against major pests of corn and cotton. Modelling studies predicted that usefulness of these resistant plants would be short-lived, as a result of the ability of insects to develop resistance against single insecticidal gene products. However, despite such dire predictions no such collapse has taken place and the acreage of transgenic insect resistance crops has been increasing at a steady rate over the 9 years since the deployment of the first transgenic insect resistant plant. However, in order to assure durability and sustainability of resistance, novel strategies have been contemplated and are being developed. This perspective addresses a number of potentially useful strategies to assure the longevity of second and third generation insect resistant plants. PMID- 16475007 TI - Homologous recombination in zebrafish ES cells. AB - Targeted insertion of a plasmid by homologous recombination was demonstrated in zebrafish ES cell cultures. Two selection strategies were used to isolate ES cell colonies that contained targeted plasmid insertions in either the no tail or myostatin I gene. One selection strategy involved the manual isolation of targeted cell colonies that were identified by the loss of fluorescent protein gene expression. A second strategy used the diphtheria toxin A-chain gene in a positive-negative selection approach. Homologous recombination was confirmed by PCR, sequence and Southern blot analysis and colonies isolated using both selection methods were expanded and maintained for multiple passages. The results demonstrate that zebrafish ES cells have potential for use in a cell-mediated gene targeting approach. PMID- 16475008 TI - Local Cre-mediated gene recombination in vascular smooth muscle cells in mice. AB - Here we describe a means to conditionally modify genes at a predefined and localized region of the vasculature using a perivascular drug delivery device (PDD). A 4-hydroxytamoxifen (4-OHT)-eluting PDD was applied around the carotid or femoral artery of a mouse strain carrying both the tamoxifen-inducible and smooth muscle cell (SMC)-specific Cre-recombinase (SM-Cre-ER(T2)) transgene and a stop floxed beta-galactosidase gene in the Rosa26 locus: the SM-CreER(T2)(ki)/rosa26 mouse. A dose and time curve of 0-10% (w/w) 4-OHT and 0-14 days application of the PDD in SM-CreER(T2)(ki)/rosa26 mice showed optimal gene recombination at 1% (w/w) 4-OHT loading at 7 days post application (carotid artery 2.4+/-1.8%; femoral artery 4.0+/-3.8% of SMCs). The unique 4-OHT-eluting PDD allowed us to achieve SMC-specific recombination in the same order of magnitude as compared to systemic tamoxifen administration. In addition, recombination was completely confined to the PDD-treated vessel wall segment. Thus, local application of a 4 OHT-eluting PDD results in vascular SMC-specific Cre-mediated recombination in SM CreER(T2)(ki)/rosa26 mice without affecting additional SMCs. PMID- 16475010 TI - Enhanced resistance to Botrytis cinerea mediated by the transgenic expression of the chitinase gene ch5B in strawberry. AB - Plants of strawberry (cultivar Pajaro) were transformed with three defense related genes: ch5B, gln2 and ap24 using Agrobacterium tumefaciens. The ch5B gene encodes for a chitinase from Phaseolus vulgaris, while gln2 and ap24 encode for a glucanase and a thaumatin-like protein, respectively, both from Nicotiana tabacum. Sixteen transgenic lines expressing one or a combination of two defense genes were obtained. Phytopathological tests showed that two transgenic lines expressing only the ch5B gene displayed high levels of resistance to gray mold disease (Botrytis cinerea). The resistance was correlated with the presence of the foreign protein CH5B and the increase of chitinolytic activity in leaves. However, resistance toward Colletotrichum acutatum, the etiological agent of the anthracnose disease, was not enhanced in the transgenic plants. These results suggest that the ch5B gene can be used to introduce transgene-mediated resistance to gray mold in strawberry, due to the lack of natural resistance to this disease in the crop. PMID- 16475009 TI - Dynamic control of oligosaccharide modification in the mammary gland: linking recombinant human erythropoietin functional analysis of transgenic mouse milk derived hEPO. AB - We analyzed two transgenic mouse lines that secrete rhEPO in their milk to assess the dynamic control of N-linked oligosaccharides. Since pharmaceutically available epoetin alpha and beta are produced in CHO cells, we compared transgenic mammary gland-derived rhEPO to its CHO cell-derived counterpart. The major glycosyltransferases that determine the N-oligosaccharides patterns of rhEPO include N-acetylglycosaminyltransferase (GnT) and alpha1,3/4 fucosyltransferase (Fuc-TIV), GnT-III, -V and Fuc-TIV expression in the mouse mammary gland is significantly higher than that in Chinese hamster ovary (CHO) derived cells, where the protein is not detectable. The data suggest that N linked sugar chain patterns of recombinant glycoproteins, produced by the mammary gland differ, since GnT-III alters the sugar pattern extensively. In our experiments, rhEPO produced by the transgenic mice contains more tetra-acidic oligosaccharide structures than epoetin alpha derived from CHO cells, a rhEPO that is widely used therapeutically. Accordingly, we examined milk-derived rhEPO activity, both in vitro and in vivo. The rhEPO protein purified from the milk of mammary glands upregulates the EPO receptor-mediated expression of the STAT5 gene in MCF-7 cells in a dose-dependent manner, similar to the effects of epoetin alpha. Furthermore, direct injection of rhEPO into the mouse tail vein leads to an increase in the levels of blood components, such as red blood cells and platelets. In light of these findings, we suggest that the mammary glands of transgenic animals provide a sufficient environment to generate rhEPO with post translational modifications for biopharmaceutical use. PMID- 16475011 TI - Evaluation of transgenic 'Chardonnay' (Vitis vinifera) containing magainin genes for resistance to crown gall and powdery mildew. AB - Magainins, short peptides with broad-spectrum antimicrobial activity in vitro, were assayed for their ability to confer resistance to pathogens in transgenic grapevines. Embryogenic cell suspensions of 'Chardonnay' (Vitis vinifera L.) were co-transformed by microprojectile bombardment with a plasmid carrying the npt-II gene and a second plasmid harboring either a natural magainin-2 (mag2) or a synthetic derivative (MSI99) gene. Magainin genes and the marker gene were driven by Arabidopsis ubiquitin-3 and ubiquitin-11 promoters, respectively. A total of 10 mag2 and 9 MSI99 regenerated lines were studied by Southern blot hybridization, which showed 1-6 transgene integration events into the plant genome. Semi-quantitative reverse transcription-polymerase chain reaction (RT PCR) revealed a variable range in transcription levels among mag2 and MSI99 lines. A positive correlation between number of integration events and transcription level was observed (p<0.05). Plants were acclimated and challenged in the greenhouse with either Agrobacterium vitis strains (bacterial crown gall pathogen) at 10(8) cfu/ml or Uncinula necator (fungal powdery mildew pathogen) at 10(5) conidia/ml for evaluation of disease resistance. A total of 6 mag2 and 5 MSI99 lines expressing the antimicrobial genes exhibited significant reductions of crown gall symptoms as compared to non-transformed controls. However, only two mag2 lines showed measurable symptom reductions in response to U. necator, but not strong resistance. Our results suggest that the expression of magainin-type genes in grapevines may be more effective against bacteria than fungi. Additional strategies to enhance transgene expression and the spectrum of resistance to grape diseases are suggested. PMID- 16475012 TI - Activation of the pathogen-inducible Gst1 promoter of potato after elicitation by Venturia inaequalis and Erwinia amylovora in transgenic apple (Malus x domestica). AB - Rather than using a constitutive promoter to drive transgenes for resistance against fungal and bacterial diseases in genetic engineering of apple (Malus x domestica) cultivars, a promoter induced only after infection was preferred. The ability of the Pgst1 promoter from potato (Solanum tuberosum L.) to drive expression of the gusA reporter gene was determined in two genotypes of apple: the fruit cultivar Royal Gala and the M.26 rootstock. beta-Glucuronidase activity in the transgenic lines grown in a growth chamber was determined quantitatively using fluorometric assays and compared to the activity in Cauliflower Mosaic Virus (CaMV) 35S promoter-driven transgenic lines. In both apple genotypes, the Pgst1 promoter exhibited a low level of expression after bacterial and fungal inoculation compared to the level obtained with the PCaMV35S promoter (15% and 8% respectively). The Pgst1 promoter was systematically activated in apple at the site of infection with a fungal pathogen. It was also activated after treatment with salicylic acid, but not after wounding. Taken together, these data show that, although the Pgst1 promoter is less active than the PCaMV35S promoter in apple, its pathogen responsiveness could be useful in driving the expression of transgenes to promote bacterial and fungal disease resistance. PMID- 16475013 TI - Production of trans-10, cis-12 conjugated linoleic acid in rice. AB - Conjugated linoleic acid (CLA) has anti-carcinogenic and anti-atherosclerosis activity, and modulatory effects on the immune system and lipid metabolism. To produce a transgenic rice plant that can accumulate CLA, a linoleate isomerase gene that can convert linoleic acid to trans-10, cis-12 CLA was introduced and expressed under the control of seed-specific promoters from the oleosin and globulin genes. The fatty acid composition of the transgenic rice grain was analyzed by gas chromatography. Although there was no clear difference in the fatty acid composition between seeds from transformed versus untransformed plants, a peak of trans-10, cis-12 CLA methyl ester, which was not present in seeds from untransformed plants, was found in transformed plants. The trans-10, cis-12 CLA comprised an average of 1.3% (w/w) of the total fatty acids in seeds carrying the oleosin promoter in comparison to 0.01% (w/w) in seeds carrying the globulin promoter. In addition, approximately 70 and 28% of the total amount of the CLA isomer were present in the triacylglycerol and free fatty acid fractions, respectively. These results demonstrate the ability to produce fatty acid components of vegetable oils with novel physiological activities in crops. PMID- 16475014 TI - Variable recombination efficiency in responder transgenes activated by Cre recombinase in the vasculature. AB - Cre recombinase has become a ubiquitous tool in transgenic strategies for regulation of transgene expression in a tissue-specific manner. We report analysis of two SM22alphaCre lines and their ability to mediate genomic recombination in five independent Cre-responsive transgenic lines. One of the SM22alphaCre lines developed was a tet-on system based on the reverse tetracycline transactivator. Our goal was to use this strategy to inhibit the Notch signaling pathway specifically in smooth muscle cells. Our responder transgenes contained a constitutively expressed marker gene (chloramphenicol acetyltransferase, CAT), flanked by loxP sites in direct orientation, upstream of Notch-related transgenes. We developed two dominant negative Notch transgenic responder lines activated by Cre-mediated DNA recombination. The first is the extracellular domain of human Jagged1, and the second is the extracellular domain of the human Notch2 receptor. Despite high expression of the marker gene in all responder lines, we found that Cre-mediated genomic recombination between these five lines was highly variable, ranging from 46 to 93% of individuals using an SM22alphaCre activating strain, or 8-58% of individuals using an inducible SM22alphartTACre. In all cases examined, detection of recombination by PCR correlated with expression of the transgene as determined by Western blot analysis. Our studies reflect the variability in recombination success based on the responder strain, presumably due to inaccessibility of the locus of integration of the responder allele. PMID- 16475015 TI - Assessment of the effectiveness of a nuclear-launched TMV-based replicon as a tool for foreign gene expression in plants in comparison to direct gene expression from a nuclear promoter. AB - An environmentally safe Tobacco Mosaic Virus (TMV)-based expression replicon was constructed that lacks movement protein (MP) and coat protein (CP), and which expresses the green fluorescent protein (GFP) gene from a full CP subgenomic promoter. The TMV replicon, whose cDNA was positioned between an enhanced Cauliflower Mosaic Virus 35S promoter (CaMV) and a self-cleaving hammerhead ribozyme with a downstream nopaline synthase gene polyadenylation signal [nos poly(A)], was assessed for its effectiveness to accumulate GFP upon agroinfiltration into plant leaves compared to a control construct in which GFP was directly expressed from the enhanced CaMV 35S promoter. It was determined that individually expressing cells produced ca. 9-fold more GFP from the TMV based replicon than from the enhanced 35S promoter. In contrast, GFP measurements from total leaf extracts determined that leaves infiltrated with the TMV-based replicon produced ca. 7-fold less GFP than the control construct. These apparently contradictory results can be explained by the low infectivity of the TMV-based replicon as it was found that the number of foci expressing GFP produced in leaves agroinfiltrated with the TMV-based replicon was ca. 66-fold lower than produced by the control. PMID- 16475018 TI - Trace minerals in experimental subarachnoid haemorrhage: zinc, copper and manganese levels in rat brain tissue, blood and urine. AB - BACKGROUND: Zinc (Zn), copper (Cu) and manganese (Mn) are involved in regulatory systems in the cell. Their role in neuromodulator activities and redox reactions has been implicated in the pathogenesis of neurological disorders. The aim of this study was to determine changes of Zn, Cu and Mn levels in brain tissue, blood and urine after experimental subarachnoid haemorrhage (SAH). The possible importance of these trace minerals on the pathogenesis of SAH was also discussed. METHOD: Rats were divided into three groups; namely a SAH group, a control group and a normal group. Blood samples in the SAH group and normal saline in the control group were injected into the cisterna magna. No surgical procedures were performed on the normal group. Brain tissue, blood and urine samples were measured for trace minerals by atomic absorption spectrophotometry. Measurements were taken on days 3, 7 and 10 after the onset in the control and SAH groups, and on the first day in the normal group. FINDINGS: The reduced blood Zn levels and increased Zn urine loss observed in the SAH group were conspicuously significant. Furthermore, significant changes in Mn levels were also seen at different stages of the trial in the SAH group. However, differences found in the Cu levels between the groups were not significant enough to explain the results. INTERPRETATION: These results suggest that the low blood Zn levels seen throughout the stages, the low brain tissue Mn levels seen during the latter part of the trial, and the low blood Mn levels observed during the early stages, may all be related to an increased risk in experimental SAH in rats. These differences may have possible role in the pathogenesis of SAH, and further investigations into the reduced blood Mn levels observed during the study may lead to new insight into the treatment of SAH. PMID- 16475017 TI - Neuroendocrine and behavioural responses to CO2 inhalation in central versus peripheral autonomic failure. AB - Multiple system atrophy (MSA) and pure autonomic failure (PAF) represent distinct pathological models of autonomic failure in humans. We have investigated the neuroendocrine, behavioural and autonomic cardiovascular responses to the 35% CO2 challenge. Nine patients with MSA, nine with PAF and five control subjects received a single breath of 35% CO2. Peripheral autonomic failure (i.e., PAF) was associated with significantly lower resting noradrenaline levels. All groups demonstrated a significant pressor response to CO2. In controls, the mean pressor response was +60.2 mm Hg, which was significantly smaller in both the PAF (+26.8 mm Hg, P < 0.01) and MSA (+18.3 mm Hg, P < 0.001) patients. In addition, the onset of the response was significantly delayed in both MSA (140.2 s) and PAF (154.2 s) patients compared with controls (32.4 s, P = 0.04 and P = 0.03, respectively). Noradrenaline levels increased only in controls. Central autonomic impairment (i.e., MSA) was associated with lower cortisol release (+8.8% in MSA compared with +35.2% in control and +23.7% in PAF) and fewer somatic symptoms of emotional arousal. Both MSA and PAF exhibit marked sympathetic autonomic impairment, however, residual (albeit differing) sympathetic pathways can still maintain a partial cardiovascular response. A central autonomic lesion, however, also appears to be associated with blunting of both cortisol and emotional responses to this stress paradigm. PMID- 16475019 TI - Combined use of a radial fore arm free flap for extra-intracranial bypass and for antero-lateral skull base reconstruction--a new technique and review of literature. AB - This article describes a new surgical technique consisting of the combined use of a fascial radial fore arm free flap (RFFF) as vascular graft for extra intracranial bypass and as dura mater plasty for reconstruction of the antero lateral skull base. This new technique is illustrated by a case of a complex intracranial meningioma with extracranial extension necessitating resection of internal carotid artery. The technical issues of antero-lateral skull base reconstruction and extra-intracranial bypass are discussed and the literature is reviewed. PMID- 16475020 TI - Subdural haemorrhage following endoscopic third ventriculostomy. A rare complication. AB - Subdural collections or hematomas are frequently observed after shunt placement [7-9, 13], but rarely after ETV [6]. A review of literature revealed 7 cases [1, 5, 6, 10, 12], of which only 1 was symptomatic [5]. We will discuss the causes, management, and methods of prevention of this complication and we will present a case of symptomatic subdural haematoma, following endoscopic third ventriculostomy for illustration. PMID- 16475022 TI - Metastatic progression of pancreatic cancer: changes in antioxidant enzymes and cell growth. AB - Pancreatic cancer has a dismal prognosis due to the fact that patients present late when metastatic disease is already present. Previous studies have demonstrated that pancreatic cancer cells have decreased levels of MnSOD, which correlates well with increased rates of tumor cell proliferation. Recently, we have found that nude mice injected with MIA PaCa-2 human pancreatic cancer cells in the flank occasionally develop ascites and intra-abdominal metastatic deposits. Mice that developed ascites were sacrificed and the ascites cultured. Necropsy demonstrated metastatic tumors in the retroperitoneum, which were excised, digested, and cultured. Western blots, enzyme activity and enzyme activity gels were performed for manganese superoxide dismutase (MnSOD), copper/zinc (CuZnSOD), catalase, and glutathione peroxidase (GPx) in the ascites cell line, metastatic tumor cell line, MIA PaCa-2 primary pancreatic cancer cell line, and the Capan-1, a metastatic pancreatic cancer cell line. Cell growth, plating efficiency, growth in soft agar and growth in nude mice were determined in the ascites, metastatic tumor, and MIA PaCa-2 cell lines. MnSOD, CuZnSOD, and GPx protein and activity were increased in the ascites, metastatic tumor, and Capan-1 cell lines compared to MIA PaCa-2. The ascites and metastatic tumor cell lines had decreased cell growth, plating efficiency, and growth in soft agar, but the ascites cell line had increased cell growth in 4 and 1% O(2) concentrations in vitro and more rapid growth in vivo. Metastatic disease is associated with changes in the content and activity of antioxidant enzymes with an associated change in growth characteristics depending on the O(2) concentrations. PMID- 16475023 TI - Chromosomal alterations in lung metastases of colorectal carcinomas: associations with tissue specific tumor dissemination. AB - Comparative genomic hybridization was used to screen colorectal carcinomas for chromosomal aberrations that are associated with the metastatic phenotype of the lung. Specimens of 13 lung metastases, 6 primary tumors, 1 lymph node metastasis, 1 liver metastasis, and 1 ovarian metastasis were investigated and added to our CGH colon cancer tumor collective, comprising 85 tumor specimens from 56 patients (see CGH online tumor database at http://amba.charite.de/cgh). Lung metastases showed more alterations than liver metastases, particularly more deletions at 1p, 3p, 9q, 12q, 17q, 19p and 22q and gains at 2q, 5p, and chromosome 6. Comparing lung metastases with their corresponding primary tumors, particularly more deletions at 3p, 8p, 12q, 17q, and 21q21 and gains at 5p were observed. Based on our results, we wish to suggest a metastatic progression model. Specific subpopulations of metastatic cells have a distinct metastatic potential, which is reflected by a non-random accumulation of chromosomal alterations. Distinct alterations already exist within the primary tumor and this "ready to go package" gives the cells the metastatic potential to achieve the complex series of events needed for metastasis. PMID- 16475024 TI - Transforming growth factor-beta1 upregulates transcription of alpha3 integrin gene in hepatocellular carcinoma cells via Ets-transcription factor-binding motif in the promoter region. AB - The invasive and metastatic potentials of hepatocellular carcinoma (HCC) are positively correlated with the expression level of alpha3beta1 integrin, a high affinity adhesion receptor for laminin isoforms. Transforming growth factor (TGF) beta1 stimulates non-invasive HCC cells to acquire invasive phenotypes in association with the enhanced expression of alpha3 integrin. In this study, we investigated the molecular mechanism underlying the upregulation of alpha3beta1 integrin by TGF-beta1 in non-invasive HepG2 HCC cells. The treatment of HepG2 cells with TGF-beta1 induced the expression of alpha3 integrin and potentiated these cells to adhere to laminin-5 and to migrate through laminin-5-coated membranes. The promoter activity was measured by luciferase assay with a series of deletion constructs of the 5'-flanking region of the mouse alpha3 integrin gene, and the results showed that the -260/-119 region (relative to the major transcription start site) contained elements responsive to TGF-beta1 stimulation. The introduction of mutations into the putative consensus binding sequence for the Ets-family of transcription factors located at -133 greatly decreased the promoter activity responding to TGF-beta1 stimulation. The nuclear proteins extracted from TGF-beta1-stimulated HepG2 cells yielded a larger amount of DNA nuclear protein complexes than did those extracted from unstimulated cells, as determined by an electrophoretic mobility shift assay using an oligonucleotide containing the Ets-site as a probe. These results suggest that TGF-beta1 stimulates HepG2 cells to express a higher level of alpha3 integrin by transcriptional upregulation via Ets transcription factors and to exhibit a more invasive phenotype. PMID- 16475025 TI - Effect of asymptomatic natural infections due to common mouse pathogens on the metastatic progression of B16 murine melanoma in C57BL/6 mice. AB - To investigate whether the presence of infections in C57BL/6 mice influences the metastatic ability of B16 melanoma (B16M) cells, we compared the susceptibility to metastasis development of pathogen-free mice with that of mice from a colony endemically infected with several mouse pathogens. We found that, compared to seronegative controls, mice that were seropositive at least to Mouse Hepatitis Virus (MHV) and Mycoplasma pulmonis: (i) exhibited a higher interindividual variability in all the parameters quantifying metastatic progression; (ii) had elevated serum levels of proinflammatory cytokines both before and at the end of the experiment; (iii) were more susceptible to hepatic metastasis. Interestingly, final levels of tumor necrosis factor (TNF)-alpha and interleukin (IL)-18 correlated with the extent of hepatic colonization by the melanoma cells. To confirm the metastasis-enhancing effect of MHV and M. pulmonis we measured the ability of B16M cells to metastasize in pathogen-free animals housed for increasing time-intervals in the vicinity of MHV(+) animals. Notably, susceptibility to metastasis was lower in animals seronegative to MHV than in MHV(+) mice, whereas the latter were less susceptible to metastasis than MHV(+) M. pulmonis(+) mice. Seropositive animals had increased levels of TNF-alpha and IL-18 suggesting that MHV and M. pulmonis enhance the metastatic ability of melanoma cells by inducing the release of proinflammatory cytokines. While our results highlight the importance of using pathogen-free animals in metastasis studies, they emphasize the need for a comprehensive health monitoring of the mice used in such studies, particularly in case of using facilities lacking appropriate containment measures. PMID- 16475026 TI - Low-dose methotrexate inhibits lung metastasis and lengthens survival in rat osteosarcoma. AB - Lung metastasis is the most crucial event affecting the treatment of osteosarcoma and is dependent on tumor angiogenesis. To improve the prognosis for patients with osteosarcoma, prevention of lung metastasis is essential. Low-dose methotrexate is a useful drug for treating a variety of diseases. Low-dose methotrexate reportedly plays a role in antiangiogenesis for the synovial blood vessels in rheumatoid arthritis. However, whether low-dose methotrexate is correlated with tumor angiogenesis and metastasis is unclear. We investigated the inhibitory effect of methotrexate on lung metastasis in a rat osteosarcoma cell line with high metastatic potential, S-SLM. Two weeks after inoculation of S-SLM cells into male Fischer 344 rats, low-dose methotrexate (1.2 mg/kg once or twice a week) or saline was intraperitonealy injected for 4 weeks and the antimetastatic effect was evaluated. Low-dose methotrexate significantly reduced the number of lung metastatic nodules and the wet weight of the lungs. Immunohistochemical staining showed a decrease in microvessel density in the metastatic nodules. We also evaluated the effect of methotrexate on the proliferation of endothelial cells and S-SLM osteosarcoma cells in vitro. Methotrexate significantly inhibited the proliferation of endothelial cells at a lower concentration than that of S-SLM osteosarcoma cells. These data suggest that low-dose methotrexate inhibited lung metastasis of osteosarcoma through its antiangiogenic activity. Our results indicate that low-dose methotrexate is a promising drug for tumor dormancy therapy in patients with osteosarcoma and lung metastasis. PMID- 16475027 TI - MUC1, MUC2, MUC4, MUC5AC and MUC6 expression in the progression of prostate cancer. AB - Molecular changes are vital for the development of prognostic markers and therapeutic modalities of prostate cancer (CaP). There is growing interest in mucins as treatment targets in human malignancies, including CaP. The role of their expression in the progression of CaP is however unclear. We examined the expressions MUC1, MUC2, MUC4, MUC5AC and MUC6 in CaP tissues using tissue microarrays (TMAs) to look for tumor-associated antigens (TAAs) for targeted therapy. In this study, 120 paraffin-embedded specimens were selected from patients who underwent radical retro-pubic prostatectomy (RRP) or trans-urethral resection of the prostate (TURP) for primary, untreated CaP and 10 matched lymph node metastases. A series of MUC monoclonal antibodies (mAbs) was used on TMAs by standard immunohistochemistry. Our results indicate that the over-expression of MUC1 was detected in 58% of primary CaP tissues and 90% of lymph node metastases but not in normal prostate or benign tissues, while the expression of MUC2, MUC4, MUC5AC and MUC6 was found to be negative in both normal and cancer tissues. Of the MUC1 positive tumors 86% were Gleason grade 7 or higher. Over-expression of MUC1 was found in late stage CaP while MUC2, 4, 5AC and 6 were negative in CaP. MUC1 is a TAA that is highly related to tumor progression in CaP patients. This antigen is ideal for targeted therapy to control micrometastases and hormone refractory disease but additional studies are necessary to assess its usefulness in patient biopsies and CaP bone metastases before clinical trial. PMID- 16475028 TI - Pre-clinical study of 213Bi labeled PAI2 for the control of micrometastatic pancreatic cancer. AB - PURPOSE: The urokinase plasminogen activator (uPA) and its receptor (uPAR) are expressed by pancreatic cancer cells and can be targeted by the plasminogen activator inhibitor type 2 (PAI2). We have labeled PAI2 with (213)Bi to form the alpha conjugate (AC), and have studied its in vitro cytotoxicity and in vivo efficacy. METHODS AND MATERIALS: The expression of uPA/uPAR on pancreatic cell lines, human pancreatic cancer tissues, lymph node metastases, and mouse xenografts were detected by immunohistochemistry, confocal microscopy, and flow cytometry. Cytotoxicity was assessed by the MTS and TUNEL assay. At 2 days post cancer cell subcutaneous inoculation, mice were injected with AC by local or systemic injection. RESULTS: uPA/uPAR is strongly expressed on pancreatic cancer cell lines and cancer tissues. The AC can target and kill cancer cells in vitro in a concentration-dependent fashion. Some 90% of TUNEL positive cells were found after incubation with 1.2 MBq/ml of AC. A single local injection of approximately 222 MBq/kg 2 days post-cell inoculation can completely inhibit tumor growth over 12 weeks, and an intraperitoneal injection of 111 MBq/kg causes significant tumor growth delay. CONCLUSIONS: (213)Bi-PAI2 can specifically target pancreatic cancer cells in vitro and inhibit tumor growth in vivo. (213)Bi-PAI2 may be a useful agent for the treatment of post-surgical pancreatic cancer patients with minimum residual disease. PMID- 16475029 TI - Endobronchial metastases from extrathoracic malignancies. AB - Endobronchial metastases (EBM) from extrapulmonary malignant tumors are rare. The most common extrathoracic malignancies associated with EBM are breast, renal and colorectal carcinomas. In this study, we aimed to evaluate the clinical, radiographic and bronchoscopic aspects of patients with EBM who were diagnosed between 1992 and 2002. Data about patients' clinical conditions, symptoms, radiographic and endoscopic findings, and histopathological examination results were investigated. EBM was defined as bronchoscopically visible lesions histopathologically identical to the primary tumor in patients with extrapulmonary malignancies. We found 15 cases with EBM. Primary tumors included breast (3), colorectal (3), and renal (2) carcinomas; Malignant Melanoma (2); synovial sarcoma (1), ampulla of Vater adenocarcinoma (1), pheochromocytoma (1), hypernephroma (1), and Hodgkin's Disease (1). The most common symptoms were dyspnea (80%), cough (66.6%) and hemoptysis (33.3%). Multiple (40%) or single (13.3%) pulmonary nodules, mediastinal or hilar lymphadenopathy (40%), and effusion (40%) were the most common radiographic findings. The mean interval from initial diagnosis to diagnosis of EBM was 32.8 months (range, 0-96 months) and median survival time was 18 months (range, 4-84). As a conclusion, various extrapulmonary tumors can metastasize to the bronchus. Symptoms and radiographic findings are similar with those in primary lung cancer. Therefore, EBM should be discriminated from primary lung cancer histopathologically. Although mean survival time is usually short, long-term survivors were reported. Consequently, treatment must be planned according to the histology of the primary tumor, evidence of metastasis to other sites and medical status of the patient. PMID- 16475031 TI - A patient's race/ethnicity does not explain the underuse of appropriate adjuvant therapy in colorectal cancer. AB - INTRODUCTION: To improve colorectal cancer outcomes, appropriate adjuvant therapy (chemotherapy, radiation therapy) should be given. Numerous studies have demonstrated underuse of adjuvant therapy in colorectal cancer. The current study examines variables associated with underuse of adjuvant therapy. METHODS: Three population-based databases were linked: California Cancer Registry, California Patient Discharge Database, 2000 Census. All colorectal cancer patients diagnosed from 1994 to 2001 were studied. Patient characteristics (age, gender, race/ethnicity, comorbidities, payer, diagnosis year, socioeconomic status) were used in five multivariate regression analyses to predict receipt of chemotherapy for Stage III colon cancer, and receipt of chemotherapy and radiation therapy for Stages II, III rectal cancer. RESULTS: The overall cohort was 18,649 Stage III colon cancer and Stages II, III rectal cancer patients. Mean age was 68.9 years, 50 percent male, 74 percent non-Hispanic white, 6 percent black, 11 percent Hispanic, 9 percent Asian, and 65 percent had no significant comorbid disease. Receipt of chemotherapy was 48 percent for Stage III colon cancer, 48 percent for Stage II rectal cancer, and 66 percent for Stage III rectal cancer. Receipt of radiation therapy was 52 percent for Stage II rectal cancer and 66 percent for Stage III rectal cancer. In all five models, low socioeconomic status predicted underuse of chemotherapy or radiation therapy (P < 0.016). Race/ethnicity was not statistically associated with underuse in any of the models. CONCLUSIONS: Most literature identifies race/ethnicity as the reason for disparate receipt of adjuvant therapy in colorectal cancer. Using a more robust database of three population-based sources, our analysis demonstrates that socioeconomic status is a more important predictor of (in)appropriate care than race/ethnicity. Explicit measures to improve care to the poor with colorectal cancer are needed. PMID- 16475030 TI - Metastasis predictive signature profiles pre-exist in normal tissues. AB - Previous studies from our laboratory have demonstrated that metastatic propensity is significantly influenced by the genetic background upon which tumors arise. We have also established that human gene expression profiles predictive of metastasis are not only present in mouse tumors with both high and low metastatic capacity, but also correlate with genetic background. These results suggest that human metastasis-predictive gene expression signatures may be significantly driven by genetic background, rather than acquired somatic mutations. To test this hypothesis, gene expression profiling was performed on inbred mouse strains with significantly different metastatic efficiencies. Analysis of previously described human metastasis signature gene expression patterns in normal tissues permitted accurate categorization of high or low metastatic mouse genotypes. Furthermore, prospective identification of animals at high risk of metastasis was achieved by using mass spectrometry to characterize salivary peptide polymorphisms in a genetically heterogeneous population. These results strongly support the role of constitutional genetic variation in modulation of metastatic efficiency and suggest that predictive signature profiles could be developed from normal tissues in humans. The ability to identify those individuals at high risk of disseminated disease at the time of clinical manifestation of a primary cancer could have a significant impact on cancer management. PMID- 16475033 TI - Our view on fissure healing should be verified. PMID- 16475032 TI - Continued survival of more than ten years, without resection of metastatic disease, in patients with metastatic colorectal cancer treated with biomodulated fluorouracil: report of two cases. AB - The treatment of metastatic colorectal cancer by chemotherapy alone was considered palliative and without the potential to cure patients unless patients were rendered resectable. We report two patients with metastatic colorectal cancer involving the liver who were considered inoperable and were treated with systemic chemotherapy using biomodulated 5-fluorouracil. Both patients received 5 fluorouracil and N-(phosphonoacetyl)-l-aspartic acid; one also received methotrexate, leucovorin, and triacetyluridine with the N-(phosphonoacetyl)-l aspartic acid and 5-fluorouracil. Both patients had a complete remission with chemotherapy and are still alive with no evidence of cancer ten years after the diagnosis of unresectable metastatic disease. These patients provide evidence that prolonged survival can be achieved withsystemic chemotherapy without the use of surgery or other forms of local therapy. These patients also confirm the importance of continued investigation of fluorouracil modulating agents, which may further enhance the recent progress made with fluorouracil-based combination chemotherapy for colorectal cancer. PMID- 16475034 TI - Colonoscopy in the office setting is safe, and financially sound ... for now. AB - PURPOSE: In 2000, the Centers for Medicare & Medicaid Services announced a plan to allow for enhanced reimbursement for office endoscopy. This change in reimbursement was phased in during three years. The purpose of this study was to evaluate the fiscal outcomes and quality measures in the first two and a one-half years of performing endoscopy in an office setting under the new Centers for Medicare & Medicaid Services guidelines. METHODS: The following financial parameters were gathered: number of endoscopies, expenses (divided into salaries and operational), net revenue, and margin for endoscopies performed in the office compared with the hospital. All endoscopies were performed by endoscopists with advanced training (gastroenterology fellowship or colon and rectal surgery residency). Monitoring equipment included continuous SaO2 and automated blood pressure in all patients and continuous electrocardiographic monitors in selected patients. Quality/safety data have been tracked in a prospective manner and include number of transfers to the hospital, perforations, bleeding requiring transfusion or hospitalization, and cardiorespiratory arrest. RESULTS: The financial outcomes are as follows: 13,285 endoscopies performed from the opening of the unit through December 2003; net revenue per case $504 per case; expense per case has dropped from $205 per case to $145 per case; the overall financial benefit of performing endoscopy in the office compared with the hospital was an additional $28 to $143 per case depending on the insurance carrier. The quality outcomes since inception of the unit include the following: 13,285 endoscopies; 0 hospital transfers, 0 cardiorespiratory arrests; 0 perforations; and 1 bleeding episode that required hospitalization. CONCLUSIONS: Endoscopy performed in the office setting is safe when done with appropriate monitoring and in the proper patient population. At the time of this study, office endoscopy also is financially rewarding but changes in Centers for Medicare & Medicaid Services reimbursement threaten the ability to retain any financial benefit. PMID- 16475036 TI - Thrombophilias--practical implications and testing caveats. AB - This review summarizes recent information about the major thrombophilic conditions, their clinical relevance, and practical aspects pertaining to testing for these thrombophilias, such as when to test and what assays are appropriate. Conditions covered include factor V Leiden, prothrombin 20210 mutation, proteins C and S, antithrombin, antiphospholipid antibodies, homocysteine, and methylene tetrahydrofolate-reductase enzyme mutation. Additional comments focus on education of patients and educational resources for patients, such as the National Alliance for Thrombosis and Thrombophilia (www.nattinfo.org). PMID- 16475037 TI - New concepts in heparin-induced thrombocytopenia: diagnosis and management. AB - Heparin-induced thrombocytopenia (HIT) is a clinicopathologic condition and adverse drug reaction caused by immunoglobulin G (IgG) antibodies directed against the heparin-platelet factor 4 complex. In most patients, the onset of thrombocytopenia begins while the patient is receiving heparin. In less than 5% of patients, the onset of thrombocytopenia begins several days following heparin discontinuation and has been termed "delayed-onset" HIT. This review summarizes the presentation and clinical course of published reports of delayed-onset HIT occurring in 30 patients. The diagnosis of delayed-onset HIT should be considered in all patients presenting with venous thromboembolism (VTE) and all patients with recent heparin exposure (within the past 14 days) who present with a low platelet count. Most patients with HIT are treated with direct thrombin inhibitors and transitioned to warfarin oral anticoagulation. Administration of direct thrombin inhibitors requires close monitoring for bleeding, dose adjustments based upon coagulation monitoring and is costly. Fondaparinux, a synthetic pentasaccharide and indirect-acting factor-Xa inhibitor, has little to no cross-reactivity with the heparin-platelet factor 4 antibody in in vitro testing. This review summarizes dosing, monitoring and outcomes of preliminary reports of fondaparinux successfully administered to 13 patients with subacute HIT and 22 patients with acute HIT. While several reports have described the treatment and prophylaxis of thrombosis in patients with HIT using fondaparinux, clinical trials should be conducted and reported before fondaparinux becomes a therapy of choice for HIT. PMID- 16475039 TI - Advances in the diagnosis of venous thromboembolism. AB - This review summarizes recent information about the diagnosis of deep venous thrombosis (DVT) and pulmonary embolism (PE) using noninvasive imaging tests, clinical assessment, and D-dimer assays, and describes how these tests can be employed in diagnostic testing algorithms for the investigation of patients with suspected DVT and PE. The clinical diagnosis of deep venous thrombosis is unreliable, but clinical prediction rules based on signs and symptoms do facilitate the categorization of patients into high, low, or medium risk categories. High sensitivity D-dimer assays further help in excluding cases but do not help in ruling in venous thromboembolism. D-dimer assays and clinical prediction rules also help in the diagnosis of pulmonary embolism. These assessments, along with objective imaging studies such as compression ultrasonography for DVT or computerized tomographic pulmonary angiograms for PE can be used in a systematic way to reliably rule in or exclude venous thromboembolism. PMID- 16475038 TI - The epidemiology of venous thromboembolism in the community: implications for prevention and management. AB - The epidemiology of venous thromboembolism (VTE) in the community has important implications for VTE prevention and management. This review describes the incidence, survival, recurrence, complications and risk factors for deep vein thrombosis and pulmonary embolism occurring in the community. VTE incidence among whites of European origin exceeds 1 per 1000; the incidence among persons of African and Asian origin may be higher and lower, respectively. VTE incidence over recent time remains unchanged. Survival after VTE is worse than expected, especially for pulmonary embolism where one-quarter of patients present as sudden death. Of those patients who survive, 30% develop VTE recurrence and venous stasis syndrome within 10 and 20 years, respectively. Common independent VTE risk factors include surgery, hospitalization for acute medical illness, nursing home confinement, trauma, active cancer, neurologic disease with extremity paresis, superficial vein thrombosis, central venous catheter/transvenous pacemaker, and among women, oral contraceptives, pregnancy and the puerperium, and hormone and SERM therapy. Exposures can identify populations at risk but have a low predictive value for the individual person. An acquired or familial thrombophilia may predict the subset of exposed persons who actually develop symptomatic VTE. In conclusion, VTE is a common, lethal disease that recurs frequently and causes serious long-term complications. To improve survival and prevent complications, VTE occurrence must be reduced. Better individual risk stratification is needed in order to modify exposures and target primary and secondary prophylaxis to the person who would benefit most. PMID- 16475042 TI - Low intensity warfarin anticoagulation is safe and effective as a long-term venous thromboembolism prevention strategy. AB - Longitudinal studies indicate a high rate of recurrence of venous thromboembolism after an episode of deep venous thrombosis or pulmonary embolism. Extended anticoagulant therapy will decrease the recurrence rate, but there is controversy as to the optimal intensity of therapy that will be effective, yet safe. The PREVENT trial addresses the question of whether long-term low intensity therapy (INR 1.5-2.0) will effectively prevent recurrence compared to placebo treatment, yet be safe without a significant increase in major bleeding. The results of this trial show a significant reduction in recurrent venous thromboembolism with a major bleeding rate that is no different than the placebo arm of the study (0.9 vs 0.4 per 100 patient years; p = 0.25). Although the ELATE trial showed greater effectiveness with no increase in bleeding in the standard intensity arm vs the low intensity arm, the question remains whether such safety can be obtained in the real world management of oral anticoagulation. PMID- 16475040 TI - The post-thrombotic syndrome: the forgotten morbidity of deep venous thrombosis. AB - The postthrombotic syndrome (PTS) is the most common complication of deep venous thrombosis (DVT) yet has received little attention from clinicians and researchers. Clinically, PTS is characterized by chronic pain, swelling, heaviness and other signs in the affected limb. In severe cases, venous ulcers may develop. PTS is burdensome and costly to patients and society because of its high prevalence, severity and chronicity. Preventing DVT with the use of effective thromboprophylaxis in high-risk patients and settings and minimizing the risk of ipsilateral DVT recurrence are likely to reduce the frequency of PTS. Compression stockings worn daily after DVT appear to reduce the incidence and severity of PTS but questions regarding their use and effectiveness remain. Future research should focus on identifying patients at high risk for PTS, assessing the role of thrombolysis in preventing PTS and evaluating the optimal use of compression stockings in preventing and treating PTS. In addition, new therapies to treat PTS should be sought and evaluated. PMID- 16475043 TI - The standard is still the standard or why an INR of 2-3 is still the optimal intensity for secondary prevention of venous thromboembolism. AB - The optimal intensity of warfarin anticoagulation for secondary prevention of venous thromboembolism is debatable. Recent studies have shed light on the issue. The two pivotal studies, ELATE and PREVENT, are reviewed and discussed. Although the ELATE and PREVENT studies offer different conclusions, the results of the two studies are consistent with each other. Low intensity warfarin is more efficacious than placebo, although it is less efficacious than standard intensity and offers no safety advantage. For long term secondary prophylaxis of spontaneous venous thromboembolism, the optimal INR intensity of warfarin remains 2.0-3.0. PMID- 16475044 TI - Anticoagulants in pregnancy. AB - Venous thromboembolic (VTE) complications are a leading cause of maternal mortality in the developed world. To reduce the incidence of VTE in pregnancy, and improve outcomes, a wider understanding of the risk factors involved and a better identification of women at risk of thrombosis coupled with effective thromboprophylaxis and treatment of VTE are required. As coumarin is unsuitable for use in pregnancy because of problems with embryopathy and risk of fetal bleeding, anticoagulation therapy in pregnancy centres on the use of low molecular-weight heparin (LMWH) and unfractionated heparin (UFH). There is now extensive experience of the safety and efficacy of LMWH in pregnancy. LMWH's, such as enoxaparin and dalteparin, have clinical and practical advantages compared with UFH in terms of improved safety (significantly lower incidence of osteoporosis and heparin induced thrombocytopenia), and patient convenience with once daily dosing for the majority of women. Such therapy is not restricted only to prevention and treatment of VTE but is now being assessed in additional clinical situations such as the prevention of pregnancy complications. PMID- 16475045 TI - New anticoagulants: anti IIa vs anti Xa--is one better? AB - Traditional anticoagulant drugs, including unfractionated heparin and warfarin, have several limitations. A new strategy for the design of new antithrombotic drugs is based on selective inhibition of a specific coagulation factor. These include direct thrombin inhibitors and factor Xa inhibitors. Two parenteral direct thrombin inhibitors, lepirudin and argatroban, have FDA approval for the management of heparin-induced thrombocytopenia. Ximelagatran, an oral prodrug of the direct thrombin inhibitor melagatran, has shown efficacy in the prevention and treatment of venous thromboembolism as well as stroke prevention in patients with atrial fibrillation. Fondaparinux is a synthetic pentasaccharide, which binds to antithrombin, thereby indirectly selectively inhibiting factor Xa. Fondaparinux has demonstrated its efficacy compared to low-molecular-weight heparin in randomized clinical trials and is FDA approved for the prevention and treatment of venous thromboembolism. A number of oral direct factor Xa inhibitors as well as other oral direct thrombin inhibitors are in clinical development for the prevention and treatment of thrombosis. PMID- 16475046 TI - Worldwide management of oral anticoagulant therapy: the ISAM study. AB - A multicenter, observational, retrospective, cross-sectional study of patients, receiving oral anticoagulation therapy (OAT) for stroke prophylaxis in chronic non-valvular atrial fibrillation (NVAF) was conducted in the US, Canada, France, Italy and Spain according to their predominant model of care [routine medical care (RMC) or Anticoagulation Clinic care (ACC)]. The study objectives were to assess anticoagulation control (time in target range), and to describe the features of the local model of care. Consecutive patients were recruited on the basis of a minimum of 60 days of oral anticoagulant treatment over a 12 month period, and clinic and physician details were captured by means of a structured face-to-face or telephone interview. Time in therapeutic range (TTR) was calculated by using linear interpolation between INR values. A total of 1511 patients were recruited, of whom 1445 were included in the analysis of TTR. TTR was higher in ACC (69.5% and 64.9% for Italy and Spain, respectively) with respect to RMC (58.1%, 62.8% and 59.3% for the US, Canada and France, respectively). Mean intervals between INR determinations were between 3 and 4 weeks. Dose changes in case of INR outside therapeutic range were more frequent in Spain and less frequent in France. Striking differences were observed in type of VKA used, specialists involved in patient management, and dosage instructions. Studying of anticoagulation management based on local models of care highlights important discrepancies among countries and suggests further standardization of the management of this important therapy is necessary. PMID- 16475048 TI - Perioperative management of oral anticoagulation: when and how to bridge. AB - The management of patients on oral anticoagulation (OAC) who need to undergo surgery or invasive procedures is problematic. "Bridging" the subtherapeutic periods with either intravenous unfractionated heparin or subcutaneous treatment dose low-molecular-weight heparin (LMWH) decreases the amount of time patients are not anticoagulated but may increase the risk of postoperative bleeding and is costly. The available literature does not provide sufficient information to allow clinicians to choose an optimal perioperative strategy. Recent studies primarily have examined the perioperative use of LMWH, and have found arterial thromboembolic rates of 0.4-1.5%. The observed incidence is greater than mathematically predicted, which may be due to a potential hypercoagulable state impacting the risk for arterial thromboembolic events. The literature suggests that major postoperative bleeding is low for invasive procedures but may be substantially higher for major surgery. Given the lack of definitive data or consensus, the decision must be based on estimates of the risks of thromboembolism and bleeding and the patient's preference. For most patients at low or moderate stroke risk, bridging will be unnecessary and may be harmful. Bridging is recommended for patients who have a high annual risk of stroke and thus have a more appreciable perioperative stroke risk. Postoperative anticoagulation must be used cautiously and patients monitored closely after major surgery due to the risk of postoperative major bleeding. PMID- 16475047 TI - The antiphospholipid syndrome: what are we really measuring? How do we measure it? And how do we treat it? AB - The antiphospholipid syndrome is described with a review of its historical development as a recognized syndrome, what constitutes an antiphospholipid antibody, how it is measured, and how the syndrome is treated. Antiphospholipid antibodies are actually antibodies to a protein, most often beta-2-glycoprotein 1, that is usually bound to a phospholipid. Some antibodies are directed towards lipid-bound prothrombin. The antibodies are measured by immunologic assays or by antibody-dependent abnormalities detected in coagulation assays. Although they prolong coagulation assays, they are associated with a thrombotic tendency rather than a bleeding disorder. There are numerous postulated mechanisms to account for the thrombotic tendency. Patients with the antiphospholipid syndrome are treated with long-term oral anticoagulation to prolong the INR to 2.0 to 3.0. For most patients, a more intense level of treatment with a higher INR is not needed. PMID- 16475049 TI - Managing oral anticoagulation requires expert experience and clinical evidence. AB - The management of patients on chronic oral anticoagulant therapy, namely Vitamin K antagonists such as warfarin, is often associated with difficult and challenging issues for the healthcare practitioner. Many of these issues, such as warfarin failure or resistance, the optimal warfarin initiation dose, the optimal target International Normalized Ratio in antiphospholipid syndrome, the optimal monitoring frequency and use of point-of-care monitoring, the management of oral anticoagulation during invasive procedures, and the management of over anticoagulation, have not been evaluated in rigorously-designed clinical trials. The latest American College of Chest Physician recommendations concerning these issues are Grade 2C, the weakest recommendations available. It remains up to the experience and expertise of the individual practitioner along with whatever clinical evidence is available in a particular healthcare environment-especially one associated with an anticoagulant management service-to implement management strategies with respect to these issues in patients on oral anticoagulation. PMID- 16475050 TI - Oral anticoagulation control: the European perspective. AB - We describe the results of a novel complete care package using point-of-care INR testing and computerized decision support for the management of oral anticoagulation by nurse-led primary care clinics in the United Kingdom. We found such therapy to be as safe and effective as traditional hospital-based care for oral anticoagulation in the UK as determined by adverse events or International Normalized Ratio (INR) time in range. We then review the literature as it pertains to the safety, efficacy, and cost effectiveness of patient self management with point-of-care INR testing at home and discuss the implications for such care in the UK. PMID- 16475052 TI - Antioxidant status and oxidative stress in professional rugby players: evolution throughout a season. AB - Physical training is known to increase the antioxidant defence system and reduce exercise-induced oxidative stress. However, intense physical aerobic and anaerobic training and competition such as those imposed on professional rugby players, can induce an increase of oxidative stress which can be implicated with the arrival of overtraining. The aim of this study was to test the effect of training and competition load on oxidative stress, antioxidant status, haematological, and cell damage markers in high-level rugby players during a competitive season. Blood samples were collected four times in one year. Oxidative stress (Rmax), antioxidant (vitamin E, uric acid, TAC, and lag phase), haematological (neutrophils and monocytes) and biochemical (CK and myoglobin) parameters, as well as training and competition load, and competition results were measured. Intense periods of training and competition (T1 and T4) induced a significant higher maximum rate of conjugated dienes oxidation (+67.2% in T1 and +40.6% in T4) compared to those observed at the reference time (T3). Those periods also induced an increase in uric acid (+6.9% and 3.2%), and inflammatory markers such as monocytes (+13.3% and 10.7%). On the other hand, vitamin E (-8.7% in T1) and lag phase (-23.0% and -14.7%) were lower during these periods showing a possible training-induced antioxidant down-regulation. The less intense period of training (T2) was accompanied by lower neutrophils (-8.5%), CK (-53.7%), and myoglobin (-16.2%) values. The results suggest that oxidative stress and antioxidant measurement are significant in the biological follow-up of athletes. PMID- 16475053 TI - Control of muscle size during disuse, disease, and aging. AB - Skeletal muscle is a highly plastic tissue. For example, muscle hypertrophies during strength training and increases its oxidative capacity in response to endurance training. Conditions associated with disuse, however, are also accompanied by adaptations, of which atrophy and a slow-to-fast transition are most prominent. Fast and slow muscles respond differently to disuse. The different response of muscle to different models of disuse reveals that loading is most important, but that also activity level, neurotrophic factors, and ageing play a part in determining the mass, morphology, contractile properties, and fatigability of a muscle. Muscle loss during disuse is a result, at least in part, of apoptosis. Finally, skeletal muscle wasting and remodelling during ageing and chronic disorders, such as chronic heart failure and chronic obstructive pulmonary disease, are not entirely attributable to disuse, but are also related to secondary consequences of the disease, most notably inflammation. Besides activating other pathways, we present evidence that inflammation during ageing and chronic disorders causes muscle wasting via alterations in abundance and/or activity of muscle specific transcription factors and induction of apoptosis, and that systemic inflammation rather than disuse is the primary cause of muscle wasting during ageing and chronic disorders. PMID- 16475054 TI - Accumulation of acetyl groups following cycling: a 1H-MR spectroscopy study. AB - Using in vivo proton magnetic resonance spectroscopy (1H-MRS), a new peak resonating at 2.13 ppm post-exercise has been attributed in the literature to the acetyl groups of acetylcarnitine. Since this peak is inconsistently generated by various submaximal exercise regimens, this study aimed at (a) verification of the previous chemical assignment, (b) determination of exercise conditions necessary for its induction, and (c) documentation of the recovery kinetics through 60 minutes following exercise. Ten healthy males (31 +/- 4 yr) cycled continuously for 45 minutes with intensity alternating between 50% (3 min) and 110% (2 min) of ventilatory threshold (VT). 1H-MR spectra were acquired from the vastus lateralis before and for 60 minutes following exercise. The peak at 2.13 ppm was not quantifiable at rest in any subject. However, it was present in all subjects following intense exercise (p < 0.0001), and expressed the chemical characteristics of an acetyl-containing compound. The estimated concentration, accumulation with high-intensity exercise, the presence as a single peak at 2.13 ppm, and the chemical shift were all consistent with the chemical and biophysical characteristics of acetyl groups associated with acetylcarnitine. This study provides further evidence that acetyl groups are robustly generated by intense exercise, and that the accumulation of acetyl groups in healthy subjects is dependent on the degree of exercise intensity. 1H-MRS may be used for the noninvasive study of muscle metabolism during exercise and recovery and may have special applications for studying the generation and transport of acetyl compounds, including acetylcarnitine. PMID- 16475055 TI - The intensity level of physical exercise and the bone metabolism response. AB - This study investigated the short-term effects of the intensity level of physical exercise on bone metabolism and related hormones. The responses of calciotropic hormones and bone biochemical markers were evaluated in seven male cyclists (mean age 24.4 years, range 20-39) during two 50-min cycling tests performed 15% below (-VT) and 15% above (+VT) the ventilatory threshold. In each test, venous blood samples were drawn at rest, at the 30th and 50th min of exercise, and after 15 min of recovery. For both intensity levels, no significant variation in calcium, 25-hydroxyvitamin D, 1.25-dihydroxyvitamin D, or cortisol level was observed. Intact parathyroid hormone (iPTH) level increased significantly after the last minute of the test (41%, p < 0.05) and peaked during the recovery (80%, p < 0.05) only in response to exercise performed at +VT. Serum phosphorus concentration rose during both tests, while albumin levels increased only at +VT. Concerning bone cell activity, osteocalcin, and type I-C telopeptide breakdown products transiently increased only in response to exercise performed at +VT (11% and 16.8%, respectively; p < 0.05). Bone alkaline phosphatase increased similarly for both intensity levels after 30 min (12%, p < 0.05) and 50 min (12% for -VT vs. 14% for +VT, p < 0.05). All markers of bone turnover returned to initial values during the recovery. In conclusion, a no-impact but intense and sustained exercise performed at +VT transiently stimulated bone turnover and iPTH secretion, suggesting the existence of a bone stimulation threshold. In addition to the well known effect of mechanical constraints, both the duration and intensity of exercise may induce changes in bone turnover. PMID- 16475057 TI - Critical velocity and lactate threshold in young swimmers. AB - The purpose of the present study was to compare the critical swimming velocity (CV) in children, with the lactate threshold (LT) and the velocity corresponding to a blood lactate concentration of 4 mmol x l(-1) (V4). Twenty swimmers (ten females and ten males, mean +/- SD age: 12.9 +/- 1.1 years, body weight: 51.2 +/- 10.0 kg, height: 157.1 +/- 9.7 cm) performed four repetitions of 200 m swimming with increasing intensity (80, 85, 90 and 100% of their 200 m maximum velocity), interspersed with 15 minutes of passive rest. Blood lactate concentration was determined after each repetition. From the speed-lactate curve, the velocity corresponding to LT and V4 was calculated. In order to calculate CV, all swimmers were timed exerting maximum effort, on distances of 50, 100, 200 and 400 m. CV was expressed as the slope of the linear relationship of time versus distance and was calculated from combinations of four (CV4) three or two timed distances. Velocity on LT (1.079 +/- 0.114 m x s(-1)) and V4 (1.106 +/- 0.112 m x s(-1)) was comparable to CV4 (1.085 +/- 0.121 m x s(-1)). CV calculated from a combination including distances of 50, 100 or 200 m were higher compared to LT (p < 0.05). V4 was higher compared to CV of 100-200-400, CV of 200-400, CV of 100-400 (p < 0.05). The CV calculated from distances of 50 and 100 m was significantly higher than all other velocities (p < 0.05). These results confirm that the critical velocity seems to be a valid, practical and time-saving, non-invasive alternative method which can be applied in the swimming pool by a coach for the evaluation of the endurance capacity of young swimmers. For practical reasons, combinations of less than four distances can be used (i.e. 50-400 m, or 50-100-400 m). PMID- 16475058 TI - Changes in the reflex excitability during and after a sustained, low-intensity muscle contraction. AB - The purpose of the study was to evaluate the soleus H-reflex amplitude during and after a low-intensity isometric contraction. Twelve healthy, untrained subjects performed a 10-minute isometric plantar flexion at 20% of their maximum voluntary contraction torque output. The electromyogram, H-reflex, and maximum M-wave (Mmax) of the soleus muscle was recorded during and 10 minutes after the end of the contraction. The results indicated that the H-reflex increased significantly (mean +/- SEM: 44.7 +/- 16.6%, p < 0.05), but when the fatigue protocol was over, the H-reflex was depressed for the first 3 minutes, relative to the H-reflex that was recorded before fatigue, when the muscle was relaxed. The Mmax did not change significantly during the whole experiment. Furthermore, the stimulation frequency (0.1 vs. 0.3 Hz) did not have any significant effect on the H-reflex modulation. The results of the current study suggest that the reflex excitability is increased as fatigue develops, whereas this increase turns to depression for the first minutes of the recovery phase. The functional significance of these changes and the neural mechanisms which might be responsible are discussed. PMID- 16475056 TI - Antioxidant status of interval-trained athletes in various sports. AB - Muscular exercise results in an increased production of free radicals and other forms of reactive oxygen species (ROS). Further, developing evidence implicates cytotoxins as an underlying etiology of exercise-induced stimuli in muscle redox status, which could result in muscle fatigue and/or injury. Two major classes of endogenous protective mechanisms (enzymatic and nonenzymatic antioxidants) work together to reduce the harmful effects of oxidants in the cell. This study examined the effects of acute physical exercise on the enzymatic antioxidant systems of different athletes and comparison was made to the mechanism of action of three main antioxidant enzymes in the blood. Handball players (n = 6), water polo players (n = 20), hockey players (n = 22), basketball players (n = 24), and a sedentary control group (n = 10 female and n = 9 male) served as the subjects of this study. The athletes were divided into two groups according to the observed changes of activity of superoxide dismutase enzyme. The antioxidant enzyme systems were characterized by catalase (CAT), glutathione-peroxidase (GPX), and superoxide-dismutase (SOD) and measured by spectrophotometry. An important finding in the present investigation is that when the activities of SOD increased, the activities of GPX and CAT increased also and this finding related to the physical status of interval-trained athletes. Positive correlation between SOD and GPX activities was observed (r = 0.38 females, r = 0.56 males; p < 0.05). We have observed that the changes in the primary antioxidant enzyme systems of athletes are sport specific, and different from control subjects. Presumably, with interval-trained athletes, hydrogen-peroxide is significantly eliminated by glutathione-peroxidase. From these results it can be concluded that the blood redox status should be taken into consideration when establishing a fitness level for individual athletes. PMID- 16475059 TI - Validity of a physical activity computer questionnaire in 12- to 18-year-old boys and girls. AB - The reliability and validity of a physical activity computer questionnaire of a usual week were studied in 33 adolescents between 12 and 18 years of age. Intraclass correlation coefficients and Kappa values were calculated to verify test-retest reliability. Validity was investigated by calculating Pearson correlation coefficients between the questionnaire and the Computer Science and Applications uniaxial accelerometer (CSA). Accelerometer data were obtained during seven successive days (sum and mean counts, estimated MET). Intraclass coefficients generally exceeded 0.70 and all Kappa values but one varied between 0.44 and 1.00. Transport variables (active transport from and to school, and during leisure time) showed no relationship with CSA. Sport participation during leisure time, sport participation summed with total transport, and the frequencies of moderate and hard activity were significantly correlated with CSA (r between 0.48 and 0.78). These data indicate that the physical activity computer variables provide reliable information. Moreover, sport participation (and summed with total transport) and the frequencies of moderate and hard activity provide valid data about adolescents' usual week physical activity, based on CSA comparison. PMID- 16475060 TI - Modelling atmospheric pollution during the games of the XXVIII Olympiad: effects on elite competitors. AB - The present study investigated the specific atmospheric conditions expected in Athens during the summer of 2004 in relation to the performance of elite athletes. DESIGN: Atmospheric pollution and weather data for the period April 16th to September 30th covering the entire greater Athens area and collected from 1984 to 2003 were used for descriptive statistics and model fitting. The analysis was focused on carbon monoxide (CO), ozone (O3), sulfur dioxide (SO2), nitrogen oxide (NO), nitrogen dioxide (NO2), and particulate matter with a diameter of < 10 microm (PM10). Factor and cluster analysis were used to describe atmospheric pollution in the northern, central, and southern sector of Athens. Generalized estimated equations (GEE) analysis was used to predict mean August 2004 pollutant concentration. Increased concentrations of O3 and PM10 (mean 2003 values: 134.3 +/- 9.3, 44 +/- 1.9 microg/m3, respectively) may generate adverse health and performance effects. The highest O3 values were recorded in the northern Athenian sector during the period June 12th to July 23rd, peaking around mid-day (12:00 18:00) (p < 0.05). The highest PM10 concentrations were recorded in the central Athenian sector during the period August 20th to September 9th, peaking at late afternoon (14:00-22:00) (p < 0.05). Similar concentrations were observed during all days of the week (p > 0.05). GEE approximated mean August 2004 pollutant concentrations similar to: CO: 2.8 (mg/m3), O3: 136, SO2: 24, NO: 134, and NO2: 106 (microg/m3). Concentrations of O3 and PM10 during the XXVIII Olympiad may generate adverse health and performance effects on the cardiovascular function of the elite competitors. (The present manuscript was submitted shortly before the start of the Games and became published after their completion. In this light, the actual pollution rates in Athens during August 2004 are presented in the Note Added in Proof as credence to the statistics used). PMID- 16475061 TI - Resistance training volume and post exercise energy expenditure. AB - The main purpose of this study was to determine if increasing the volume of a resistance training program, while keeping the intensity the same would significantly increase the recovery energy expenditure, as measured by the excess post-exercise energy expenditure (EPEE). Fifteen resistance-trained women completed two resistance training protocols. Both protocols consisted of nine exercises performed to volitional fatigue, at the previously determined eight RM. The only difference between the two protocols was the number of sets (1-set or 3 sets). All subjects completed both protocols in a randomized and counterbalanced fashion. Energy expenditure, as determined by oxygen consumption and respiratory exchange ratio (RER), was measured continuously prior to exercise, during the exercise session, and during the entire 120 min of recovery. Fingertip blood lactate concentration was determined at baseline, immediately post-exercise, and every 30 min in recovery. The net energy expenditure during exercise was significantly greater during the 3-set protocol (661.9 +/- 43.9 kj) than the 1 set protocol (234.7 +/- 13.4 kj). However, per minute of exercise, there was no significant difference between the two protocols. During the 120 min of recovery, the net EPEE was not significantly different between the two protocols (93.3 +/- 20.1 kj following the 1-set protocol, and 94.1 +/- 7.1 kj following the 3-set protocol). However, when comparing the EPEE to the number of minutes spent in the exercise session, the 1-set protocol led to a significantly greater energy expenditure in the recovery period (1.0 +/- 0.22 kj/min) than did the 3-set protocol (0.4 +/- 0.03 kj/min). These data indicate that in previously trained women, increasing the volume of resistance training, from 1-set to 3-sets, while keeping the intensity the same will not significantly increase EPEE above what is found in a lower volume of work. PMID- 16475062 TI - Oxygen uptake kinetics during supra VO2max treadmill running in humans. AB - Accurate classification of VO2 kinetics is essential to correctly interpret its control mechanisms. The purpose of this study was to examine VO2 kinetics in severe and supra-maximal intensity running exercise using two modelling techniques. Nine subjects (mean +/- S.D: age, 27 +/- 7 years; mass, 69.8 +/- 9.0 kg; VO2max, 59.1 +/- 1.8 mL x kg x min(-1)) performed a series of "square-wave" exercise transitions to exhaustion at running speeds equivalent to 80% of the difference between the VO2 at LT and VO2max (delta), and at 100%, 110% and 120% VO2max. The VO2 response was modelled with an exponential model and with a semi logarithmic transformation, the latter assuming a certain steady state VO2. With the exponential model there was a significant reduction in the "gain" of the primary component in supra-maximal exercise (167 +/- 5 mL x kg(-1) x km(-1) at 80% delta to 142 +/- 5 mL x kg(-1) x km(-1) at 120% VO2max, p = 0.005). The time constant of the primary component also reduced significantly with increasing intensity (17.8 +/- 1.1 s at 80% delta to 12.5 +/- 1.2 s at 120% VO2max, p < 0.05). However, in contrast, using the semi-log model, the time constant significantly increased with intensity (30.9 +/- 13.5 s at 80% delta to 72.2 +/- 23.9 s at 120% VO2max, p < 0.05). Not withstanding the need for careful interpretation of mathematically modelled data, these results demonstrate that neither the gain nor the time constant of the VO2 primary component during treadmill running are invariant across the severe and supra-maximal exercise intensity domains when fit with an exponential model. This suggests the need for a reappraisal of the VO2/work rate relationship in running exercise. PMID- 16475063 TI - Spring-mass model characteristics during sprint running: correlation with performance and fatigue-induced changes. AB - Spring-mass model properties of eight non-specialized male runners were measured during four straight 100-m sprints on an athletics track. A recently developed simple measurement method allowed to calculate leg and vertical stiffness, vertical displacement of the center of mass, and stride temporal characteristics. Changes in these mechanical parameters were studied and correlated with those of sprint performance. During the first 100 m, forward velocity showed significant variations (mean value of 8.10 +/- 0.31 m x s(-1) over the entire 100-m), while leg and vertical stiffnesses (19.5 +/- 4.3 kN x m(-1) and 93.9 +/- 12.4 kN x m( 1), respectively) remained constant. No significant link was found between mechanical and performance parameters over this first sprint. During the following three sprints, vertical stiffness, step frequency, and contact time significantly decreased (20.6 +/- 7.9%, 8.03 +/- 3.34%, and 14.7 +/- 7.2% of the first 100-m value, respectively) with decreasing maximal and mean velocities (10.9 +/- 2.0% and 7.30 +/- 5.23%, respectively), whereas leg stiffness and maximal force remained constant. Furthermore, changes between these mechanical and performance parameters were significantly related, showing the clear relationship between impairment in spring-mass model properties of the runners' lower limbs and the decrease in performance in fatigue conditions induced by the repetition of these all-out efforts. PMID- 16475064 TI - Is aerobic power really critical for success in alpine skiing? PMID- 16475066 TI - [Eight characteristics of leaders in crisis management]. AB - Everything is changing in our daily life. The ancient Chinese philosopher, Sun Tzu, said "The art of war teaches us to rely not on the likelihood of the enemy's not coming, but on our own readiness to receive him". In the challenging environment of today, nursing administrators should prepare themselves by developing a range of skills to face the changing world. This article introduces Chinese and Western leadership theories and suggests eight key characteristics of successful leaders in health care. Equipped with these characteristics, nursing leaders should be able to collaborate with other professionals in their organization to manage any crisis. Success in crisis management also enhances an organization's general potential to succeed in this competitive world. PMID- 16475067 TI - [Crises of trust]. AB - Extensive media coverage is warning of a crisis of trust that has emerged as a serious issue in our society. This article explores the meaning of "crisis," concepts of crisis management, mechanisms for building trust, and the underlying significance of trust and distrust. Evidence is adduced to testify to the erosion of trust and factors in our society that reflect the potential for crises of trust. Organizational decision makers, including hospital managers, are urged to identify and reflect upon weaknesses in their organizations in order that remedial action can be taken to preempt such crises. PMID- 16475068 TI - [The challenge facing nursing education: to develop clinical performance examination strategies in nursing practice]. AB - The clinical performance examinations of qualified health professionals have become a central professional and corporate issue. Nursing experts have to face trends in nursing care delivery toward a clinical competence-based care model that assesses the professional skills and efficacy of nursing professional and students. Objective, structured clinical examination standards and strategies can be effective vehicles for nursing education and practice, promoting the mastery of clinical nursing skills and fostering an evidenced-based practice approach to clinical care. PMID- 16475069 TI - [Crisis management in clinical nursing practice: a case analysis]. AB - The clinical medical environment is highly changeable. Hospital administrators must face and resolve various kinds of crisis. Effective crisis management can preempt or reduce negative impacts on an organization, as well as reduce unnecessary costs and bolster an organization's reputation. This article uses actual cases to describe crisis management with reference to four stages of crisis development and 6M models, providing instruction from experience, Educational growth, and enhanced crisis handling and crisis management skills. PMID- 16475070 TI - [Cases discussed of crisis management]. AB - Everything undergoes structural shifts from day to day. Any kind of change can become not only a distinguishing feature, but also a source of crisis. The only way to respond to this is to maintain crisis awareness, since crises can develop faster than we imagine. Sound preparation for any manager should therefore involve planning to deal with a crisis that might arise at any time. A crisis could be a turning point for a positive form of change; this will depend on maturity of attitude and sound familiarity with the four stages of crisis management. In each stage, we should consider whether urgent and rapid action might truly create a positive opportunity, or actually make the situation worse. This article discusses the cost of errors in crisis management and the benefits of prompt, positive action in relation to the different examples established by four well-known cases. It is hoped that these will benefit nursing managers by improving crisis management skills in clinical practice. PMID- 16475071 TI - [A project to provide instruction in the nursing of elderly patients with total hip replacement (T.H.R.)]. AB - This project was aimed at improving the nursing of patients who have undergone total hip replacements. Investigation showed the following problems with existing nursing instruction in this area: lack of standard instruction, outdated educational materials, a 33.75% rate of completion of instruction lack of familiarity with instruction materials, and an average satisfaction score of 2.56 among nurses who have undergone instruction; The reading for patient's satisfaction with the guidance of nurses was 2.04. After site investigation, status analysis and reference check, we proposed the following program. (1) Establish standards and monitor tools for instruction for nursing total hip replacement patients, including "Caring standard", "Guidance for nursing instruction", "Nursing instruction sheet", "Notes at nursing instruction", "Satisfaction scoring system for nursing instruction"; (2) Carry out a training course to enhance nursing staff's knowledge about caring for patients with total hip replacement. After program had been implemented, a completion rate of 88.56% was achieved, and the satisfaction scores among nursing staff and patients were 4.3 and 4.36 respectively. This result shows that when we undertake reform at various different levels--including systemic structure, processing and monitoring -this can radically improve the quality of nursing instruction. PMID- 16475072 TI - [A pilot study: gastric motility and nausea/vomiting in two leukemia children receiving chemotherapy]. AB - The purpose of the study was to identify the association between chemotherapy induced nausea/vomiting and changes to the electrogastrogram (EGG) of two children suffering from leukemia. After receiving written consent/assent, the children, both with acute lymphoblastic leukemia (ALL), were recruited. One of the subjects, a ten year-old boy, was given 1.1 gm Cytarabine (intravenous infusion for six hours per day) for three days and Tropisetron 5 mg intravenous infusion for 24 hours. The other subject, an eight year-old girl, received the induction phase of TPOG 93HR chemotherapy, which included Epirubicin, Vincristin, L-asparaginase, and Prednisolone and Tropisetron 5 mg on Day 1. The EGG recordings of both patients were recorded for a total of 42 hours by cutaneous electrogastrography over a seven day period. This included two-hour and four-hour readings taken before and immediately following the administration of chemotherapy each day. The position, movements, and activities of the children while on the EGG were recorded on digital video. Four episodes of nausea and vomiting were detected during this period. Pre- and post-nausea and vomiting during the EGG were analyzed using spectrum analysis after the deletion of motion artifacts. The findings of this study indicated that two episodes of nausea were 5.3-10.3% bradygastria and 2.1-10.3% tachygastria, with 85.8% and 100% normal gastric slow waves detected by EGG during the pre-vomiting period. Tachygastria was present in 3.4% and 12.2% of the post-vomiting period of each episode. The association of artifacts with position, movement, and activities must be considered during data collection. PMID- 16475073 TI - [Crisis: a concept analysis]. AB - When patients and their families face the sudden impact of disease, a crisis may ensue. Nursing staff are in the best position to prevent and manage the crisis situation; but, this concept has not been well developed in the nursing field in Taiwan. The purpose of this paper is to apply the methodology outlined by Walker and Avant (1995) to analyze the concept of crisis. The steps include a review of the literature on conceptual definitions of crisis, determining the defining attributes, constructing model, borderline, contrary, and related cases, and identifying antecedents and consequences. Finally, the empirical referents of crisis are explored. It is hoped that this analysis can help nursing staff to gain a better understanding of the concept of crisis and then to apply the knowledge gained to clinical practice. PMID- 16475074 TI - [Risk discourse in Hormone Replacement Therapy]. AB - The medical world has used Hormone Replacement Therapy (HRT) on menopausal women for over 70 years, but controversy surrounding the treatment remains. Findings reported by the NIH, U.S.A. in July 2002 indicated that HRT might cause cardiovascular and cancer risks, and resulted in serious concern among menopausal women using HRT in Taiwan. One after the other, members of the medical profession in Taiwan have argued that the medicine used in the research was different (from that used in Taiwan) and that the constitutions of western women are different from those of Taiwanese women, attempting to limit the negative impact of this research. HRT is one of the most widely used, but also one of the most controversial of medical technologies. The technico-scientific and sociocultural perspectives are two major approaches to the conceptualization of risk in relation to technology. Based on cognitive science, the technico-scientific discourse of risk emphasizes human forecasting of harmful events and scientific measures for preventing risk. The sociocultural perspective emphasizes that the risk is socially constructed and applies to control deviance. In addition, surveillance and regulated measures are applied to high-risk populations, as normalizing technology with moral implications. This paper suggests that health care professionals examine how HRT users interpret the meanings of risk. Reflection on the process and criteria for identifying high-risk populations is also suggested. Partnership with and empowerment of HRT users is advocated in preference to the use of regulating and normalizing measures. PMID- 16475075 TI - [A two-stage translation and test the validity and reliability of a foreign instrument]. AB - Examination of the reliability and validity of research tools is an important process in nursing research. As all scales are developed on the basis of a specific set of concepts or application to a specific population, great care must be taken in the translation and handling of questionnaires developed in different languages or, in particular, by people from different cultures. Only after passing through an integral process of repetitive translation and reliability/validity examinations can they be regarded as accurate research tools and be widely utilized. This study exercises Brislin's revised Translation Model to overcome cross-cultural differences, and also uses a new research tool developed overseas, the Diabetes Management Self-Efficacy Scale (DMSES), as an example for explaining the two-stage design of the translation and reliability/validity process. The First Stage is the translation stage, which involves forward translation, consensus meetings, backward translation, and review by potential users. The Second Stage involves psychometric analysis, which includes validity and reliability testing. Such careful processes should reveal the problems encountered when adopting foreign scales for domestic use, and thereby increase the "nativeness" and adaptability of these scales. PMID- 16475076 TI - [Critical social theory perspective on respite care service in Taiwan]. AB - The temporary rest provided by respite care both benefits caregiver well-being and allows care receivers to continue living in the community, effectively reducing rates of institutional placement. The necessity and effectiveness of respite care are well documented in the literature. However, only a few care providers take advantage of such services. Previous research, which has focused largely on personal factors contributing to low utilization rates, limits our understanding of the situation and of the constraints not related to personal factors. This paper attempts to move beyond the individual perspective by framing research in critical social theory in order to understand the current social, political and economical factors that limit caregiver utilization of respite services. Critical social theory, a reflection theory, emphasizes empowerment and conscious-raising in order to promote control of oppressed subjects. In applying critical theory, this paper encourages nurses to raise caregivers' consciousness of their constraining circumstances in order to empower them to access respite services on their own volition. PMID- 16475077 TI - [Nursing experience for a cancer recurrence patient in puberty]. AB - This case report discusses the impact on the mind and body of a cancer recurrence patient in puberty, as he comes to terms with, and adjusts to, a recrudescence in his illness. In our study of the nursing process, we used Gorden's assessment tools for health patterns and gathered subjective or objective information through interviews and observations to establish case problems (i.e., "changes in body and mind", "personal maladjustments related to changes" "injuries of high risk" and "oral cavity mucosa ulcers", etc.). After gaining an understanding of the previous medical treatments employed in this case, in a way of respecting the medical selection and considering the developmental task at the stage of life, we provided support and encouragement through appropriate communications and expressions of sympathy with the objective of enhancing patient self-caring and capacity for adjustment in order to help the patient achieve the best possible quality of life. PMID- 16475078 TI - [The adaptation process and behavioral responses of a patient with peripheral arterial occlusive disease who has undergone amputation]. AB - The purpose of this study was to explore the adaptation and behavioral responses of an 85 year-old woman who suffered from gangrene related to peripheral arterial occlusive disease and finally underwent lower limb amputation. The field method was adopted. Records of the patient's behaviors were collected by observation and interviews. The data were recorded by nursing process recording and analyzed using the Behavior Classification Model. The findings showed the patient's behavioral responses to be categorized by three phases. The first phase was the impact phase: patient objected to amputation strongly because it would destroy her body image. She experienced moods of anxiety and denial moods etc. The second phase was the regressive phase: after the amputation, the patient experienced a sense of loss and discomfort while having to cooperate with medical treatment and rehabilitation. Silence, withdrawal, and despair were some of her reactions. The third phase was the acceptance/ reconstruction phase: the patient accepted the fact of her amputation with time and started her rehabilitation. During the process of providing nursing care, we helped the patient to vent her emotional responses, helped her to develop awareness of her ability to face the loss of her limb, and aggressively planned individualized rehabilitation for her. Finally, these interventions enabled the patient to overcome the impact of her amputation. The findings of this study should provide references for further clinical nursing care. PMID- 16475079 TI - A life-history perspective on short- and long-term consequences of compensatory growth. AB - Compensatory or catch-up growth (CG) is widely observed following periods of resource deprivation. Because of this commonness, it is generally assumed that compensatory growth is adaptive, but most theory to date has explicitly ignored considerations of fitness. Following a period of deprivation, when resources become plentiful again, individuals may not respond at all and continue on a "normal" trajectory from a smaller size at age, may exhibit faster-than-normal growth immediately following the end of the period, or may adopt a growth strategy that involves faster-than-normal growth at some later time. Compensating individuals may also overtake control individuals who have been growing normally throughout. We hypothesize that the key to understanding CG is that growth leads to the accumulation of damage at the cellular level that is expressed (and thus must be modeled) at the level of the organism. We show that a life-history model incorporating the mortality consequences of both size and damage provides a framework for understanding compensatory growth. We use the theory to classify physiological and life-history characteristics for which CG is predicted to be the optimal response to deprivation. PMID- 16475080 TI - Ontogenetic patterns in heritable variation for body size: using random regression models in a wild ungulate population. AB - Body size is an important determinant of fitness in many organisms. While size will typically change over the lifetime of an individual, heritable components of phenotypic variance may also show ontogenetic variation. We estimated genetic (additive and maternal) and environmental covariance structures for a size trait (June weight) measured over the first 5 years of life in a natural population of bighorn sheep Ovis canadensis. We also assessed the utility of random regression models for estimating these structures. Additive genetic variance was found for June weight, with heritability increasing over ontogeny because of declining environmental variance. This pattern, mirrored at the phenotypic level, likely reflects viability selection acting on early size traits. Maternal genetic effects were significant at ages 0 and 1, having important evolutionary implications for early weight, but declined with age being negligible by age 2. Strong positive genetic correlations between age-specific traits suggest that selection on June weight at any age will likely induce positively correlated responses across ontogeny. Random regression modeling yielded similar results to traditional methods. However, by facilitating more efficient data use where phenotypic sampling is incomplete, random regression should allow better estimation of genetic (co)variances for size and growth traits in natural populations. PMID- 16475081 TI - Symmetry breaking in escaping ants. AB - The phenomenon of herding is a very general feature of the collective behavior of many species in panic conditions, including humans. It has been predicted theoretically that panic-induced herding in individuals confined to a room can produce a nonsymmetrical use of two identical exit doors. Here we demonstrate the existence of that phenomenon in experiments, using ants as a model of pedestrians. We show that ants confined to a cell with two symmetrically located exits use both exits in approximately equal proportions to abandon it in normal conditions but prefer one of the exits if panic is created by adding a repellent fluid. In addition, we are able to reproduce the observed escape dynamics in detail using a modification of a previous theoretical model that includes herding associated with a panic parameter as a central ingredient. Our experimental results, combined with theoretical models, suggest that some features of the collective behavior of humans and ants can be quite similar when escaping under panic. PMID- 16475082 TI - Effects of mechanical stress and plant density on mechanical characteristics, growth, and lifetime reproduction of tobacco plants. AB - Plastic increases in stem elongation in dense vegetation are generally believed to be induced by canopy shading, but because plants protect each other from wind, shielding (reduced mechanical stress) could also play a role. To address this issue, tobacco Nicotiana tabacum plants were subjected to two levels of mechanical stress, 0 (control) or 40 (flexed) daily flexures, and grown solitarily, in a dense monostand (with plants of only one mechanical treatment), or in a mixed stand (flexed and control plants grown together). Flexed plants produced shorter and thicker stems with a lower Young's modulus than control plants, while dense-stand plants had relatively taller and thinner stems than solitary ones. Flexing effects on stem characteristics were independent of stand density. Growth, reproduction, and survival of solitary plants were not affected by flexing, while in the monostand growth was slightly reduced. But in the mixed stand, flexed plants were readily shaded by controls and had considerably lower growth, survival, and reproduction rates. These results suggest that wind shielding indeed plays a role in the plastic increase in stem elongation of plants in dense vegetation and that this response can have important consequences for competitive ability and lifetime seed production. PMID- 16475083 TI - A large cost of female mate sampling in pronghorn. AB - We measured the energy cost of mate sampling by female pronghorn (Antilocapra americana), a species for which there are no apparent direct benefits of mate choice and for which the sampling tactic most closely resembles best-of-n or comparative Bayes. We used Global Positioning System collars to record the position of individuals at 10-min intervals during the 2 weeks preceding estrus in females that actively sampled and in females that did not sample. The difference in the 2-week energy costs of these two classes of females was 8,200 (+/-2,300) kJ, or roughly one-half of the energy cost of a single day. This value, expressed as the fraction of total yearly energy expenditure, is 59 times the value reported for a lekking bird. Our finding calls into question the common assumption in models of mate search that the cost of search is negligible as well as the common assumption that the cost of sampling must be small when there are only indirect benefits of female choice. PMID- 16475084 TI - When can herbivores slow or reverse the spread of an invading plant? A test case from Mount St. Helens. AB - Here we study the spatial dynamics of a coinvading consumer-resource pair. We present a theoretical treatment with extensive empirical data from a long-studied field system in which native herbivorous insects attack a population of lupine plants recolonizing a primary successional landscape created by the 1980 volcanic eruption of Mount St. Helens. Using detailed data on the life history and interaction strengths of the lupine and one of its herbivores, we develop a system of integrodifference equations to study plant-herbivore invasion dynamics. Our analyses yield several new insights into the spatial dynamics of coinvasions. In particular, we demonstrate that aspects of plant population growth and the intensity of herbivory under low-density conditions can determine whether the plant population spreads across a landscape or is prevented from doing so by the herbivore. In addition, we characterize the existence of threshold levels of spatial extent and/or temporal advantage for the plant that together define critical values of "invasion momentum," beyond which herbivores are unable to reverse a plant invasion. We conclude by discussing the implications of our findings for successional dynamics and the use of biological control agents to limit the spread of pest species. PMID- 16475085 TI - Mitochondrial uncoupling proteins: new perspectives for evolutionary ecologists. AB - Reactive oxygen species (ROS)-induced damage on host cells and molecules has been considered the most likely proximal mechanism responsible for the age-related decline in organismal performance. Organisms have two possible ways to reduce the negative effect of ROS: disposing of effective antioxidant defenses and minimizing ROS production. The unbalance between the amount of ROS produced and the availability of antioxidant defenses determines the intensity of so-called oxidative stress. Interestingly, most studies that deal with the effect of oxidative stress on organismal performance have focused on the antioxidant defense compartment and, surprisingly, have neglected the mechanisms that control ROS production within mitochondria. Uncoupling proteins (UCPs), mitochondrial transporters of the inner membrane, are involved in the control of redox state of cells and in the production of mitochondrial ROS. Given their function, UCPs might therefore represent a major mechanistic link between metabolic activity and fitness. We suggest that by exploring the role of expression and function of UCPs both in experimental as well as in comparative studies, evolutionary biologists may gain better insight into this link. PMID- 16475086 TI - Correcting for regression to the mean in behavior and ecology. AB - If two successive trait measurements have a less-than-perfect correlation, individuals or populations will, on average, tend to be closer to the mean on the second measurement (the so-called regression effect). Thus, there is a negative correlation between an individual's state at time 1 and the change in state from time 1 to time 2. In addition, whenever groups differ in their initial mean values, the expected change in the mean value from time 1 to time 2 will differ among the groups. For example, birds feeding nestlings lose weight, but initially heavier birds lose more weight than lighter birds, a result expected from the regression effect. In sexual selection, males who remain unmated in the first year are, on average, less attractive than mated males. The regression effect predicts that these males will increase their attractiveness in the second year more than mated males. In well-designed experiments, changes in the experimental and control groups would be compared. In observational studies, however, no such comparison is available, and expected differential effects must be accounted for before they can be attributed to external causes. We describe methods to correct for the regression effect and assess alternative causal explanations. PMID- 16475087 TI - Inbreeding depression and the evolution of dispersal rates: a multilocus model. AB - Inbreeding depression is one of the possible reasons organisms disperse. In this article, we present a two-locus model for the evolution of dispersal in the presence of inbreeding depression. The first locus codes for a modifier of the migration rate, while the second locus is a selected locus generating inbreeding depression. We express the change in frequency of the migration modifier as a function of allele frequencies and genetic associations and then use a quasi equilibrium assumption to express genetic associations as functions of allele frequencies. Our model disentangles two effects of inbreeding depression: it gives an advantage to migrant individuals because their offspring are on average less homozygous, but it also decreases the degree of population structure, thus decreasing the strength of kin selection for dispersal. We then extend our model to include an infinite number of selected loci. When the cost of dispersal is not too high, the model predictions are confirmed by multilocus simulation results and show that inbreeding depression can have a substantial effect on the dispersal rate. For high costs of dispersal, we observe discrepancies between the model and the simulations, probably caused by associations among selected loci, which are neglected in the analysis. PMID- 16475088 TI - Acquired immunity and stochasticity in epidemic intervals impede the evolution of host disease resistance. AB - Disease can generate intense selection pressure on host populations, but here we show that acquired immunity in a population subject to repeated disease outbreaks can impede the evolution of genetic disease resistance by maintaining susceptible genotypes in the population. Interference between the life-history schedule of a species and periodicity of the disease has unintuitive effects on selection intensity, and stochasticity in outbreak period further reduces the rate of spread of disease-resistance alleles. A general age-structured population genetic model was developed and parameterized using empirical data for phocine distemper virus (PDV) epizootics in harbor seals. Scenarios with acquired immunity had lower levels of epizootic mortality compared with scenarios without acquired immunity for the first PDV outbreaks, but this pattern was reversed after about five disease cycles. Without acquired immunity, evolution of disease resistance was more rapid, and long-term population size variation is efficiently dampened. Acquired immunity has the potential to significantly influence rapid evolutionary dynamics of a host population in response to age-structured disease selection and to alter predicted selection intensities compared with epidemiological models that do not consider such feedback. This may have important implications for evolutionary population dynamics in a range of human, agricultural, and wildlife disease settings. PMID- 16475089 TI - Combining stochastic models with experiments to understand the dynamics of monarch butterfly colonization. AB - Stochastic models are of increasing importance in ecology but are usually only applied to observational data. Here we use a stochastic population model to combine experimental and observational data to understand the colonization of old fields by monarch butterflies Danaus plexippus. We experimentally tested for density dependence in oviposition rates when predators were excluded, and we measured predation rates under natural conditions. Significance tests on the resulting data showed that both oviposition and predation were density dependent but could not show how oviposition and mortality combine to determine egg densities in nature. We therefore used our data to calculate the Akaike Information Criterion to choose between a nested suite of stochastic models that differed in their oviposition and mortality terms. When we simply fit the models to the observational data, the best model assumed density independence in both oviposition and predation. When we instead first estimated the oviposition rate at low density from experimental data, however, the best model included density dependence in oviposition, and a model that included density dependence in both oviposition and predation performed nearly as well. This result is consistent with our experiments and suggests that experiments can enhance the usefulness of stochastic models in ecology. PMID- 16475090 TI - Stress hormones: a link between maternal condition and sex-biased reproductive investment. AB - In species where offspring fitness is sex-specifically influenced by maternal reproductive condition, sex allocation theory predicts that poor-quality mothers should invest in the evolutionarily less expensive sex. Despite an accumulation of evidence that mothers can sex-specifically modulate investment in offspring in relation to maternal quality, few mechanisms have been proposed as to how this is achieved. We explored a hormonal mechanism for sex-biased maternal investment by measuring and experimentally manipulating baseline levels of the stress hormone corticosterone in laying wild female European starlings (Sturnus vulgaris) and examining effects on sex ratio and sex-specific offspring phenotype adjustment. Here we show that baseline plasma corticosterone is negatively correlated with energetic body condition in laying starlings, and subsequent experimental elevation of maternal baseline plasma corticosterone increased yolk corticosterone without altering maternal condition or egg quality per se. Hormonal elevation resulted in the following: female-biased hatching sex ratios (caused by elevated male embryonic mortality), lighter male offspring at hatching (which subsequently grew more slowly during postnatal development), and lower cell-mediated immune (phytohemagglutinin) responses in males compared with control-born males; female offspring were unaffected by the manipulation in both years of the study. Elevated maternal corticosterone therefore resulted in a sex biased adjustment of offspring quality favorable to female offspring via both a sex ratio bias and a modulation of male phenotype at hatching. In birds, deposition of yolk corticosterone may benefit mothers by acting as a bet-hedging strategy in stochastic environments where the correlation between environmental cues at laying (and therefore potentially maternal condition) and conditions during chick-rearing might be low and unpredictable. Together with recent studies in other vertebrate taxa, these results suggest that maternal stress hormones provide a mechanistic link between maternal quality and sex-biased maternal investment in offspring. PMID- 16475091 TI - Explaining Dioscorides' "double difference": why are some mushrooms poisonous, and do they signal their unprofitability? AB - The adaptive significance of toxins in mushrooms has received very little consideration, although it is clear that poisons have appeared (and/or disappeared) many times in mushrooms' evolutionary history. One possibility is that poisons have evolved in some mushroom species to deter their consumption by would-be fungivores before spore dispersal. If this is so, then one might expect poisonous mushrooms to signal their unprofitability in some way. In this study, we have conducted the first formal analysis of the ecological and morphological traits associated with edible and poisonous mushrooms in North America and Europe. Poisonous mushrooms do not tend to be more colorful or aggregated than edible mushrooms, but they are more likely to exhibit distinctive odors even when phylogenetic relationships are accounted for. This raises the intriguing possibility that some poisonous species of mushrooms have evolved warning odors (and perhaps tastes) to enhance avoidance learning by fungivores. PMID- 16475092 TI - Genomic adaptation to acidic environment: evidence from Helicobacter pylori. AB - The origin of new functions is fundamental in understanding evolution, and three processes known as adaptation, preadaptation, and exaptation have been proposed as possible evolutionary pathways leading to the origin of new functions. Here we examine the origin of an acid resistance mechanism in the mammalian gastric pathogen Helicobacter pylori, with reference to these three evolutionary pathways. The mechanism involved is that H. pylori, when exposed to the acidic environment in mammalian stomach, restricts the acute proton entry across its membrane by its increased usage of positively charged amino acids in the inner and outer membrane proteins. The results of our comparative genomic analysis between H. pylori, the two closely related species Helicobacter hepaticus and Campylobacter jejuni, and other relevant proteobacterial species are incompatible with the hypotheses invoking preadaptation or exaptation. The acid resistance mechanism most likely arose by selection favoring an increased usage of positively charged lysine in membrane proteins. PMID- 16475093 TI - Superparasitism evolution: adaptation or manipulation? AB - Superparasitism refers to the oviposition behavior of parasitoid females who lay their eggs in an already parasitized host. This often yields intense competition among larvae that are sharing the same host. Why would a female oviposit in such hostile habitat instead of looking for a better quality, unparasitized host? Here we present a continuous-time model of host-parasitoid interaction and discuss alternative scenarios. This model is first used to analyze the evolution of the superparasitism behavior of a solitary proovigenic parasitoid under both time and egg limitation. Then, following the recent discovery by Varaldi et al., we allow the parasitoid to be infected by a virus that alters the superparasitism behavior of its host to enhance its own horizontal transmission. The analysis of the coevolution of this manipulative behavior with the oviposition behavior of uninfected females clarifies and quantifies the conflict that emerges between the parasitoid and its virus. The model also yields new testable predictions. For example, we expect that uninfected parasitoids should superparasite less after coevolving with the manipulative virus. More generally, this model provides a theoretical framework for analyzing the evolution of the manipulation of parasitoid life-history traits by microparasites. PMID- 16475094 TI - Ontogeny of additive and maternal genetic effects: lessons from domestic mammals. AB - Evolution of size and growth depends on heritable variation arising from additive and maternal genetic effects. Levels of heritable (and nonheritable) variation might change over ontogeny, increasing through "variance compounding" or decreasing through "compensatory growth." We test for these processes using a meta-analysis of age-specific weight traits in domestic ungulates. Generally, mean standardized variance components decrease with age, consistent with compensatory growth. Phenotypic convergence among adult sheep occurs through decreasing environmental and maternal genetic variation. Maternal variation similarly declines in cattle. Maternal genetic effects are thus reduced with age (both in absolute and relative terms). Significant trends in heritability (decreasing in cattle, increasing in sheep) result from declining maternal and environmental components rather than from changing additive variation. There was no evidence for increasing standardized variance components. Any compounding must therefore be masked by more important compensatory processes. While extrapolation of these patterns to processes in natural population is difficult, our results highlight the inadequacy of assuming constancy in genetic parameters over ontogeny. Negative covariance between direct and maternal genetic effects was common. Negative correlations with additive and maternal genetic variances indicate that antagonistic pleiotropy (between additive and maternal genetic effects) may maintain genetic variance and limit responses to selection. PMID- 16475095 TI - Convergent evolution and divergent selection: lizards at the White Sands ecotone. AB - Ecological transition zones, where organismal phenotypes result from a delicate balance between selection and migration, highlight the interplay of local adaptation and gene flow. Here, I study the response of an entire species assemblage to natural selection across a common ecotone. Three lizard species, distributed along a dramatic environmental gradient in substrate color, display convergent adaptation of blanched coloration on the gypsum dunes of White Sands National Monument. I investigate the role of gene flow in modulating phenotypic response to selection by quantifying color variation and genetic variation across the ecotone. I find species differences in degree of background matching and in genetic connectivity of populations across the ecotone. Differences among species in phenotypic response to selection scale precisely to levels of genetic isolation. Species with higher levels of gene flow across the ecotone exhibit less dramatic responses to selection. Results also reveal a strong signal of ecologically mediated divergence for White Sands lizards. For all species, phenotypic variation is better explained by habitat similarity than genetic similarity. Convergent evolution of blanched coloration at White Sands clearly reflects the action of strong divergent selection; however, adaptive response appears to be modulated by gene flow and demographic history and can be predicted by divergence-with-gene-flow models. PMID- 16475096 TI - Spatiotemporal dynamics in marginal populations. AB - Population dynamics across a mortality gradient at an ecological margin are investigated using a novel modeling approach that allows direct comparison of stochastic spatially explicit simulation results with deterministic mean field models. The results show that demographic stochasticity has a large effect at population margins such that density profiles fall off more sharply than predicted by mean field models. Substantial spatial structure emerges at the margin, and spatial correlations (measured parallel to the margin) exhibit a sharp maximum in the tail of the density profile, indicating that spatial substructuring is greatest at an intermediate point across the ecological gradient. Such substructuring may have a substantial impact on Allee effects and evolutionary processes in marginal populations. PMID- 16475097 TI - A cognitive framework for mate choice and species recognition. AB - Mating decisions contribute to both the fitness of individuals and the emergence of evolutionary diversity, yet little is known about their cognitive architecture. We propose a simple model that describes how preferences are translated into decisions and how seemingly disparate patterns of preference can emerge from a single perceptual process. The model proposes that females use error-prone estimates of attractiveness to select mates based on a simple decision rule: choose the most attractive available male that exceeds some minimal criterion. We test the model in the tungara frog, a well-characterized species with an apparent dissociation between mechanisms of mate choice and species recognition. As suggested by our model results, we find that a mate attraction feature alters assessments of species status. Next, we compare female preferences in one-choice and two-choice tests, contexts thought to emphasize species recognition and mate choice, respectively. To do so, we use the model to generate maximum-likelihood estimators of preference strengths from empirical data. We find that a single representation of preferences is sufficient to explain response probabilities in both contexts across a wide range of stimuli. In this species, mate choice and species recognition are accurately and simply summarized by our model. While the findings resolve long-standing anomalies, they also illustrate how models of choice can bridge theoretical and empirical treatments of animal decisions. The data demonstrate a remarkable congruity of perceptual processes across contexts, tasks, and taxa. PMID- 16475098 TI - Can environmental variation generate positive indirect effects in a model of shared predation? AB - Classic models of apparent competition predict negative indirect effects between prey with a shared enemy. If predator per capita growth rates are nonlinear, then endogenously generated periodic cycles are predicted to generate less negative or even positive indirect effects between prey. Here I determine how exogenous mechanisms such as environmental variation could modify indirect effects. I find that exogenous variation can have a broader range of effects on indirect interactions than endogenously generated cycles. Indirect effects are altered by environmental variation even in simple models for which the per capita growth rate of the predator species is a linear function of population densities. Temporal variation that affects the predator attack rate or the conversion efficiency can lead to large increases or decreases in the indirect effects between prey, dependent on how prey populations co-vary with the environmental variation. Positive indirect effects can occur when the period of environmental variation is close to the natural period of the biological system and shifts in subharmonic resonance occur with the addition of the second prey. Models that include nonlinear numerical responses generally lead to indirect effects that are sensitive to environmental variation in more parameters and across a wider range of frequencies. PMID- 16475099 TI - Alternative designs and the evolution of functional diversity. AB - According to conventional wisdom, functional diversity is exclusively a consequence of species having evolved adaptations to fill different niches within a heterogeneous environment. This view anticipates only one optimal combination of trait values in a given environment, but it is also conceivable that alternative designs of equal fitness in the same environment might evolve. To investigate that possibility, we use a genetic algorithm to search for optimal combinations of 34 functional traits in a realistic model of tree seedling growth and survival. We show that separate lineages of seedlings evolving in identical environments result in many alternative functional designs of approximately equal fitness. PMID- 16475100 TI - The role of local species abundance in the evolution of pollinator attraction in flowering plants. AB - We present a population genetic model that incorporates aspects of pollinator efficiency and abundance to examine the effect of the local plant community on the evolution of floral trait specialization. Our model predicts that plant species evolve to be pollinator specialists on the most effective and common pollinators when their abundance is low relative to other plant species in the community (i.e., conspecific pollen is relatively rare) and evolve to be pollinator generalists when they are numerically dominant (i.e., conspecific pollen is abundant). Strong flower constancy also favors generalist floral traits. Furthermore, generalist species are predicted to differentiate when there is a concave trade-off in attracting pollinator species with different floral trait preferences. This result implies that populations that evolve toward a generalist strategy may be more prone to speciation. Ours is the first theoretical model to include local species abundance explicitly, despite the fact that it has been previously identified as an important factor in the evolution of plant specialization. Our results add a layer of ecological complexity to previous models of floral evolution and therefore have the potential to improve our power to predict circumstances under which specialized and generalized plant pollinator interactions should evolve. PMID- 16475101 TI - The evolution of resource specialization through frequency-dependent and frequency-independent mechanisms. AB - Levins's fitness set approach has shaped the intuition of many evolutionary ecologists about resource specialization: if the set of possible phenotypes is convex, a generalist is favored, while either of the two specialists is predicted for concave phenotype sets. An important aspect of Levins's approach is that it explicitly excludes frequency-dependent selection. Frequency dependence emerged in a series of models that studied the degree of character displacement of two consumers coexisting on two resources. Surprisingly, the evolutionary dynamics of a single consumer type under frequency dependence has not been studied in detail. We analyze a model of one evolving consumer feeding on two resources and show that, depending on the trait considered to be subject to evolutionary change, selection is either frequency independent or frequency dependent. This difference is explained by the effects different foraging traits have on the consumer resource interactions. If selection is frequency dependent, then the population can become dimorphic through evolutionary branching at the trait value of the generalist. Those traits with frequency-independent selection, however, do indeed follow the predictions based on Levins's fitness set approach. This dichotomy in the evolutionary dynamics of traits involved in the same foraging process was not previously recognized. PMID- 16475102 TI - Cryptic sexual conflict in gift-giving insects: chasing the chase-away. AB - The chase-away model of sexual selection posits that elaborate male sexual displays arise because they exploit preexisting biases in females' sensory systems and induce females to mate in a suboptimal manner. An essential element of this hypothesis is that such manipulation should quickly lead to female resistance to male displays. Nuptial food gifts may be a frequent conduit by which males attempt to influence the mating behavior of females against females' own reproductive interests. In decorated crickets Gryllodes sigillatus, such inducements come in the form of a spermatophylax, a gelatinous mass forming part of the male's spermatophore and consumed by the female after mating. We conducted experiments in which spermatophylaxes obtained from male G. sigillatus were offered as novel food gifts to females of a non-gift-giving species (Acheta domesticus) having no evolutionary history of spermatophylax consumption. Female A. domesticus that were allowed to consume the spermatophylax took significantly longer to remate than when given no such opportunity. In contrast, when female G. sigillatus were prevented from consuming their partners' nuptial gifts, there was no difference in their propensity to remate relative to females permitted to consume a food gift after mating. These results suggest that the spermatophylax synthesized by male G. sigillatus contains substances designed to inhibit the sexual receptivity of their mates but that female G. sigillatus have evolved reduced responsiveness to these substances. PMID- 16475103 TI - Imbalance of predator and prey armament: geographic clines in phenotypic interface and natural selection. AB - The escalation of defensive/offensive arms is ubiquitous in prey-predator evolutionary interactions. However, there may be a geographically varying imbalance in the armaments of participating species that affects the outcome of local interactions. In a system involving the Japanese camellia (Camellia japonica) and its obligate seed predator, the camellia weevil (Curculio camelliae), we investigated the geographic variation in physical defensive/offensive traits and that in natural selection on the plant's defense among 17 populations over a 700-km-wide area in Japan. The sizes of the plant defensive apparatus (pericarp thickness) and the weevil offensive apparatus (rostrum length) clearly correlated with each other across populations. Nevertheless, the balance in armaments between the two species was geographically structured. In the populations for which the balance was relatively advantageous for the plant's defense, natural selection on the trait was stronger because in the other populations, most plant individuals were too vulnerable to resist the attacks of the weevil, and their seeds were infested independent of pericarp thickness. We also found that the imbalance between the defensive/offensive armaments and the intensity of natural selection showed clear latitudinal clines. Overall, our results suggest that the imbalance of armament between sympatric prey and predator could determine the strength of local selection and that climatic conditions could affect the local and overall trajectory of coevolutionary arms races. PMID- 16475104 TI - Seed banks, salmon, and sleeping genes: effective population size in semelparous, age-structured species with fluctuating abundance. AB - Previous studies reached contrasting conclusions regarding how fluctuations in abundance affect Ne in semelparous species with variable age at maturity: that Ne is determined by the arithmetic mean N among the T years within a generation (Ne approximately = T(N)t; monocarpic plants with seed banks) or the harmonic mean (Ne approximately T[symbol: see text]; Pacific salmon). I show that these conclusions arise from different model assumptions rather than inherent differences between the species. Sequentially applying standard, discrete generation formulas for inbreeding Ne to a series of nominal generations accurately predicts the multigenerational rate of increase in inbreeding. Variability in mean realized reproductive success across years (kt) is the most important factor determining Ne and Ne/N. When abundance is driven by random variation in kt, Ne < or = T[symbol: see text] < T(N)t. With random variation in Nt and constant per capita seed production (C), variation in kt is low and Ne approximately T[symbol: see text]; however, if C varies among years, Ne can be closer to T[symbol: see text]. Because population regulation affects the genetic contribution of entire cohorts of monocarpic perennials, Ne for these species may be more closely approximated by T[symbol: see text] than by T(N)t. With density dependent compensation, Cov(kt, Nt) < 0, and Ne is further reduced because relatively few breeders make a disproportionate contribution to the next generation. PMID- 16475105 TI - Female disdain for swords in a swordtail fish. AB - Studies of mate choice evolution tend to focus on how female mating preferences are acquired and how they select for greater elaboration of male traits. By contrast, far less is known about how female preferences might be lost or reversed. In swordtail fish Xiphophorus, female preference for the sword ornament is an ancestral trait. Xiphophorus birchmanni, however, is one species that has secondarily lost the sword. Using synthetic animation playback of "virtual" males, we found that female X. birchmanni preferred a swordless conspecific over a sworded heterospecific. Moreover, when offered the choice between a conspecific without a sword and one with a digitally attached sword, females preferred the former. These results suggest female preferences need not always select for elaboration of male traits, and they provide a plausible explanation for the lack of introgression of a sexual trait in a naturally occurring hybrid zone. PMID- 16475106 TI - Hydrolases in the hypopharyngeal glands of workers of Scaptotrigona postica and Apis mellifera (Hymenoptera, Apinae). AB - Hydrolytic enzymes from hypopharyngeal gland extracts of newly emerged, nurse and foraging workers of two eusocial bees, Scaptotrigona postica, a native Brazilian stingless bee, and the Africanized honey bee (Apis mellifera) in Brazil, were compared. The hypopharyngeal gland is rich in enzymes in both species. Fifteen different enzymes were found in the extracts, with only a few quantitative differences between the species. Some of the enzymes present in the extracts may have intracellular functions, while others seem to be digestive enzymes. Scaptotrigona postica, had lower beta-glucosidase and higher lipase esterase activities than A. mellifera. The differences may be due to different feeding habits and behavioral peculiarities of the two species. PMID- 16475107 TI - Characters that differ between diploid and haploid honey bee (Apis mellifera) drones. AB - Diploid males have long been considered a curiosity contradictory to the haplo diploid mode of sex determination in the Hymenoptera. In Apis mellifera, 'false' diploid male larvae are eliminated by worker cannibalism immediately after hatching. A 'cannibalism substance' produced by diploid drone larvae to induce worker-assisted suicide has been hypothesized, but it has never been detected. Diploid drones are only removed some hours after hatching. Older larvae are evidently not regarded as 'false males' and instead are regularly nursed by the brood-attending worker bees. As the pheromonal cues presumably are located on the surface of newly hatched bee larvae, we extracted the cuticular secretions and analyzed their chemical composition by gas chromatograph-mass spectrometry (GC MS) analyses. Larvae were sexed and then reared in vitro for up to three days. The GC-MS pattern that was obtained, with alkanes as the major compounds, was compared between diploid and haploid drone larvae. We also examined some physical parameters of adult drones. There was no difference between diploid and haploid males in their weight at the day of emergence. The diploid adult drones had fewer wing hooks and smaller testes. The sperm DNA content was 0.30 and 0.15 pg per nucleus, giving an exact 2:1 ratio for the gametocytes of diploid and haploid drones, respectively. Vitellogenin was found in the hemolymph of both types of imaginal drones at 5 to 6 days, with a significantly lower titer in the diploids. PMID- 16475108 TI - The use of RAPD to characterize Bipolaris sorokiniana isolates. AB - Bipolaris sorokiniana is a phytopathogenic fungus causing diseases of cereal crops such as common root rot, the leaf spot disease, seedling blight, and black point of the grain. Random-amplified polymorphic DNA (RAPD) assay was used to investigate the genetic diversity of 20 isolates collected from different cultivars in wheat-producing regions in Brazil. Seventy primers, with random nucleotide sequences, were tested. Reproducibility to amplify the genomic DNA of isolates was found for 30 of the 70 primers tested, generating between 1 and 17 fragments ranging from 0.35 to 2.0 kb (average size). The degree of similarity between samples was calculated through simple association and the dendrogram was assessed using the unweighted pair group method with arithmetical average. After the RAPD analyses 19 isolates were closely grouped, having a similarity coefficient of >or= 78%. Isolate I017 showed very low similarity coefficients, ranging between 38 and 46%. The RAPD analyses provided important information as to the degree of genetic variability and the relationship between the isolates investigated, revealing polymorphism and establishing electrophoretic profiles useful to characterize the phytopathogen. PMID- 16475109 TI - Cytogenetic and genotoxic effects of the insecticides, imidacloprid and methamidophos. AB - We examined the cytogenetic and genotoxic effects of the neonicotinoid insecticide imidacloprid and the organophosphate insecticide methamidophos, when administered alone or in combination. These insecticides were tested with the bone marrow chromosome aberration assay and micronucleus test in rats and by the bacterial mutation assay (Salmonella/microsome mutagenicity assay). Wistar albino rats were orally fed daily with laboratory chow treated with various concentrations of insecticides, 50 and 100 mg/kg imidacloprid, 2.5 and 5 mg/kg methamidophos, and 2.5 and 5 mg/kg imidacloprid plus methamidophos, respectively, for 90 days. Numerical and structural chromosomal aberrations were evaluated. Significant differences were detected between all the insecticide-administered groups versus the control group and between the two concentrations of the pesticide-treated groups. Both concentrations of the insecticides induced a dose related increase in the micronucleus frequency (P < 0.05). Dose-related increases in the number of revertants were observed with the two Salmonella strains (TA98 and TA100). All tested doses of the insecticides demonstrated mutagenic activity in the presence of S9 mix. These results lead us to the conclusion that the synergistic effect of methamidophos and imidacloprid causes an increase in potential damage to non-target organisms. PMID- 16475110 TI - Karyotypic characterization of Iheringichthys labrosus (Pisces, Pimelodidae): C-, G- and restriction endonuclease banding. AB - Various chromosomal banding techniques were utilized on the catfish, Iheringichthys labrosus, taken from the Capivara Reservoir. C-banding regions were evidenced in telomeric regions of most of the chromosomes. The B microchromosome appeared totally heterochromatic. The restriction endonuclease AluI produced a banding pattern similar to C-banding in some chromosomes; the B microchromosome, when present, was not digested by this enzyme and remained stained. G-banding was conspicuous in almost all the chromosomes, with the centromeres showing negative G-banding. When the restriction endonuclease BamHI was used, most of the telomeres remained intact, while some centromeres were weakly digested. The B chromosome was also not digested by this enzyme. The first pair of chromosomes showed a pattern of longitudinal bands, both with G-banding and BamHI; this was more evident with G-banding. This banding pattern can be considered a chromosomal marker for this population of I. labrosus. PMID- 16475111 TI - Molecular phylogeny of penaeid shrimps inferred from two mitochondrial markers. AB - Penaeid shrimps are an important resource in crustacean fisheries, representing more than the half of the gross production of shrimp worldwide. In the present study, we used a sample of wide-ranging diversity (41 shrimp species) and two mitochondrial markers (758 bp) to clarify the evolutionary relationships among Penaeidae genera. Three different methodologies of tree reconstruction were employed in the study: maximum likelihood, neighbor joining and Bayesian analysis. Our results suggest that the old Penaeus genus is monophyletic and that the inclusion of the Solenocera genus within the Penaeidae family remains uncertain. With respect to Metapenaeopsis monophyly, species of this genus appeared clustered, but with a nonsignificant bootstrap value. These results elucidate some features of the unclear evolution of Penaeidae and may contribute to the taxonomic characterization of this family. PMID- 16475113 TI - Karyotype of Brazilian Anopheles albitarsis sensu lato (Diptera:Culicidae). AB - Anopheles (Nyssorhynchus) albitarsis sensu lato is an important malaria vector in Brazil, especially in the Brazilian Amazon region. Chromosome preparations of fourth-instar larvae of A. albitarsis from Iranduba and Coari (AM) and Ilha Comprida (SP) were analyzed for karyotype determination and to improve cytogenetic identification of this species. Anopheles albitarsis possesses 2n = 6 chromosomes, with two pairs (submetacentric and metacentric) of autosomes and one pair of sex chromosomes, with X-Y dimorphism. The sex pair is homomorphic and acrocentric in females and heteromorphic in males, with a punctiform Y chromosome. Somatic pairing was detected in the prometaphase and metaphase chromosomes of the three A. albitarsis populations. Apparently, sex chromosome evolution in the Culicidae does not function as does evolution in the Culicidae, since it occurs in the subfamily Anophelinae, which possesses heteromorphic sex chromosomes and is regarded as primitive, based on several criteria. These karyotype data on the albitarsis complex reinforce the hypothesis that sex chromosome evolution in the subfamily Anophelinae is conserved, and the variation revealed in the mean size of chromosomes in three populations indicates that selective pressure in these populations is occurring only at a genetic level. PMID- 16475112 TI - Heterochromatin and chromosome evolution: a FISH probe of Cebus apella paraguayanus (Primate: Platyrrhini) developed by chromosome microdissection. AB - Neotropical Primate karyotypes are highly variable, particularly in the heterochromatic regions, not only regarding the amount of heterochromatin, but also the composition. G and C banding and FISH techniques provide useful information to characterize interspecific relationships. We used chromosome microdissection to develop a FISH probe of the chromosome 11 heterochromatic block (11qHe+) of Cebus apella paraguayanus (CAPp). Fragments of the 11qHe+ microdissected from fibroblast cell culture were collected in a PCR tube, amplified by degenerate oligonucleotide primer-PCR and subsequently labeled. The specificity of the FISH probe was confirmed in metaphases of some Ceboidea species. Signals were located in the He+ of chromosomes 4, 11, 12, 13, and 19 of CAPp and in the He+ of chromosomes 4, 12 and 13 of C. a. nigritus (CAPn); no signals were observed when other Ceboidea species were analyzed. We propose that the heterochromatin observed in CAPp and CAPn is specific for these species. We consider this C. apella heterochromatin identity as a possible key for the interpretation of chromosomal evolution in these Ceboidea. PMID- 16475115 TI - Localization of rDNA sites in holocentric chromosomes of three species of triatomines (Heteroptera, Triatominae). AB - Chromatin organization in the holocentric chromosomes of three triatomines species was cytologically studied by fluorescent in situ hybridization with a 45S rDNA probe of Drosophila melanogaster to localize ribosomal genes. In Triatoma tibiamaculata, metaphases I showed telomeric highlights in a single, larger bivalent. In T. protacta, hybridization was detected in one of the telomeres of an autosomal chromosome. In T. platensis, there were highlights in a single, smaller chromosome (X chromosome). The results obtained did not agree with the expected localization of rDNA genes in the sex chromosomes of triatomines, as demonstrated by silver impregnation, and suggest that the chromosome reorganization that occurred in this group during evolution may be a more important mechanism involved in rDNA distribution. PMID- 16475114 TI - Association among oxidized LDL levels, MnSOD, apolipoprotein E polymorphisms, and cardiovascular risk factors in a south Brazilian region population. AB - Oxidized LDL (ox-LDL) is involved in the initiation and progression of atherosclerosis. Many factors can affect the LDL oxidation such as oxidative stress. The present study tested whether ox-LDL levels would be associated with apolipoprotein E (APOE), manganese superoxide dismutase (MnSOD) Ala16Val polymorphisms, and classic cardiovascular risk factors. ox-LDL levels were measured by thiobarbituric acid-reactive substances and both polymorphisms were determined by polymerase chain reaction/restriction fragment length polymorphism in a sample of 252 subjects (70 men, 182 women, mean age, 54-85 years). Subjects with ox-LDL >or=0.5 nmol/mg apoprotein were considered the high level group (HLG, N = 82) and subjects with ox-LDL <0.5 nmol/mg apoprotein were considered the expected level group (ELG, N = 170). Classic risk factors were also evaluated. The results showed that diabetes mellitus was more prevalent in HLG, whereas other cardiovascular risk factors were similar between groups. The APOE genotype frequencies did not differ between HLG and ELG subjects. However, AA genotype from MnSOD polymorphism was more frequent in ELG (chi(2) = 8.48; P = 0.014). AV and VV subjects from ELG present highest ox-LDL levels (OR = 3.61; CI95% = 1.42 9.17) than AA. Additional analysis did not find gene-gene interactions associated with ox-LDL levels. Multivariate analysis showed that diabetes and the MnSOD polymorphism were independent factors associated with higher ox-LDL levels in HLG. The results suggest that an important framework on modulation of the redox status influenced by genetic polymorphisms could affect the cardiovascular homeostasis. PMID- 16475116 TI - Breeding cassava for apomixis. AB - Apomixis genes have been successfully transferred to cassava (Manihot esculenta) by hybridizing it with the wild species, M. glaziovii. The interspecific hybrid of cassava and M. glaziovii was exposed to open pollination during three subsequent generations. Seven sibs and the maternal progenitor of the fourth generation were genotyped using five microsatellite loci previously developed for cassava. All sibs were identical with each other and with their maternal progenitor. Sibs from M. glaziovii proved to be identical when examined by the same microsatellite loci. This evidence leads to the conclusion that apomixis does occur in wild-cassava relatives and apparently has played an important role in Manihot speciation. This is the first report of nearly 100% apomixis. PMID- 16475117 TI - Applicability of RAPD markers on silver-stained polyacrylamide gels to ascertain genetic diversity in Peripatus acacioi (Peripatidae; Onychophora). AB - RAPD (random amplification of polymorphic DNA) molecular markers can be utilized for analyzing genetic variability in populations for which only a few or no molecular markers are available. They were used in a study of an endangered species, Peripatus acacioi, found in the Tripui Ecological Station, in Ouro Preto, MG, Brazil. The ecological station was specifically created to protect this velvet worm species, the first of this group found in Brazil. For an initial evaluation of the genetic diversity of this species, DNA samples from the lobopods of four individuals, collected at random, were analyzed using RAPD. Each reaction was run with a different primer (Operon RAPD 10-mer Kits), totaling 13 primers (OPC2, OPC3, OPC4, OPC6, OPC8, OPC10, OPC11, OPL2, OPL7, OPL11, OPL13, OPL18, and OPL19). Due to the low amplification yield, RAPD fragments were separated in polyacrylamide gels and stained with silver nitrate. Numerous bands were observed. Fifty-five of the amplified bands proved to be reproducible, both in terms of presence and intensity. Among these, 27 were variable and 28 were constant. The average number of bands per gel was 4.2. Nine of the 13 primers tested allowed the identification of constant and variable bands among these four individuals. RAPD analysis of genetic variation using silver-stained polyacrylamide gel electrophoresis provided measures of band sharing among the individuals, and therefore could be used in population genetics studies of P. acacioi. PMID- 16475118 TI - Development and optimization of a fluorescent differential display PCR system for studying bovine embryo development in vitro. AB - Differential display is a widely used methodology to identify genes that are differentially expressed in biological samples. We developed a new protocol for the amplification and recovery of differentially expressed genes from extremely small initial amounts of RNA (10 to 25 pg mRNA) from preimplantation bovine embryos. The cDNAs generated with an anchor primer, associated with a universal sequence, were amplified with an arbitrary primer and a single fluorescently labeled primer. Amplification products were easily visualized with a fluorescence scanner, without the need for radioisotopes. Nineteen isolated fragments were cloned and sequenced, confirming the expected primer sequences and allowing the recognition and identification of gene transcripts involved in bovine embryonic physiology. PMID- 16475119 TI - First chromosome data on Steindachneridion scripta (Pisces, Siluriformes, Pimelodidae) from Brazilian rivers: Giemsa, CBG, G-, and RE banding. AB - A cytogenetic study was performed on the large pimelodid species Steindachneridion scripta (Siluriformes, Sorubiminae) from the Parana River basin (Brazil). Chromosome preparations were obtained avoiding sacrifice of the specimens, by means of lymphocyte culture, and several staining and chromosome banding techniques were applied. The karyotype consisted of 56 chromosomes, 24 metacentrics, 20 submetacentrics, 4 subtelocentrics, and 8 acrocentrics (fundamental number = 104). The first pair of acrocentric chromosomes (pair 25) consistently had a decondensed secondary constriction; the C-banding pattern of some chromosomes allows them to be considered cytogenetic markers (i.e., pairs 1, 3, 4, 6, 7, 9, 13, 23, and 24). G-banding and restriction enzymes provided patterns that helped improve chromosome pairing. This is the first report on a Neotropical pimelodid species of economic interest using several cytogenetic techniques and providing an integral karyotypic characterization. PMID- 16475120 TI - Multiple Mantel tests and isolation-by-distance, taking into account long-term historical divergence. AB - Mantel tests of matrix correspondence have been widely used in population genetics to examine microevolutionary processes, such as isolation-by-distance (IBD). We used partial and multiple Mantel tests to simultaneously test long-term historical effects and current divergence and equilibrium processes, such as IBD. We used these procedures to calculate genetic divergence among Eugenia dysenterica (Myrtaceae) populations in Central Brazil. The Nei's genetic distances between pairs of local populations were strongly correlated with geographic distances, suggesting an IBD process, but field observations and the geographic distribution of the samples suggest that populations may have been subjected to more complex evolutionary processes of genetic divergence. Partial Mantel regression was used to partition the effects of geographic structure and long-term divergence associated with a possible historical barrier. The R(2) of the model with both effects was 73.3%, and after the partition 21.9% of the variation in the genetic distances could be attributed to long-term historical divergence alone, whereas only 1.5% of the variation in genetic distances could be attributed to IBD. As expected, there was a large overlap between these processes when explaining genetic divergence, so it was not possible to entirely partition divergence between historical and contemporary processes. PMID- 16475121 TI - A three-step molecular protocol employing DNA obtained from dried blood spots for neonatal screening for 45,X Turner syndrome. AB - Turner syndrome (TS) is one of the most common human chromosomal abnormalities; it is characterized by the presence of one normal X chromosome and the complete or partial loss of the second X chromosome. The early recognition of TS patients allows for adequate therapy for short stature and pubertal sex steroid substitution. We developed a cost-effective molecular diagnostic tool that can be used to identify 45,X TS patients from dried blood spots, for possible use in neonatal screening for TS. We used a three-step method for 45,X TS detection: i) DNA extraction from dried blood spot samples, ii) pre-PCR HpaII digestion (methylation-sensitive enzyme) and iii) GeneScan analysis of selected cases. DAX 1 gene amplification was used to recognize DNA integrity, and the androgen receptor gene (Xq11-12), which is both a highly polymorphic and methylated gene, was used to determine the number of X chromosome alleles. Using this three-step diagnostic procedure, we detected apparent TS in 1/304 (0.33%) samples; such individuals should be submitted to clinical examination and karyotype confirmation. The three-step 45,X TS neonatal screening protocol is a simple, reliable, fast (under 30 h) and cost-effective diagnostic tool, useful for the neonatal detection of TS. PMID- 16475122 TI - Long survival in a 69,XXX triploid infant in Greece. AB - The live birth of a triploidy infant is a very rare event and death usually occurs within the first hours of life. Triploid cases with a survival of more than two months are infrequent. We report on an infant with a 69,XXX chromosome constitution who survived 164 days. Chromosomal analysis demonstrated a 69,XXX karyotype with no evidence of mosaicism. This is the longest survival reported for this condition to date in Greece and the fourth longest worldwide. The infant was admitted to our clinic several times due to respiratory problems, and supplementary oxygen was required. The improved survival of our case was possibly due to better management of respiratory illness and prematurity, and these are essential factors that physicians should consider carefully with such rare cases. PMID- 16475123 TI - Genetic trends of abdominal fat content in a male broiler chicken line. AB - Data of chickens from a broiler-breeding program were collected and used to determine the genetic trends of absolute and relative abdominal fat content. The genetic trends were estimated by the regression of trait genetic value averages on hatch-years. Genetic values from 32,485 individuals were used for regression analysis. The genetic trend estimate for absolute abdominal fat content was +0.39 g per year, indicating that abdominal fat deposition in the analyzed line, in absolute terms, tended to increase, making the existing excess fat deposition in the broilers even worse. However, the genetic trend of relative abdominal fat content was not significant, indicating that there is no increase on abdominal fat content when it is corrected for body weight. PMID- 16475124 TI - Composition of constitutive heterochromatin of Pseudonannolene strinatii Mauries, 1974 (Diplopoda, Spirostreptida) analyzed by AT/CG specific fluorochromes. AB - Triple staining with fluorochromes (DA/DAPI/CMA) and C-banding were used to characterize the composition of Pseudonannolene strinatii heterochromatin. C banding showed C+ bands of different labeling intensity on chromosomes 1 and 2 in some cells. Fluorochrome staining revealed DAPI+ regions corresponding to the C banding pattern, indicating that the heterochromatin of this species is abundant in AT-rich sequences. PMID- 16475125 TI - DNA repair gene polymorphisms and susceptibility to familial breast cancer in a group of patients from Campinas, Brazil. AB - Several studies have reported that the genes involved in DNA repair and in the maintenance of genome integrity play a crucial role in protecting against mutations that lead to cancer. Epidemiologic evidence has shown that the inheritance of genetic variants at one or more loci results in a reduced DNA repair capacity and in an increased risk of cancer. Polymorphisms have been identified in several DNA repair genes, such as XRCC1, XPD, XRCC3, and RAD51, but the influence of specific genetic variants on repair phenotype and cancer risk has not yet been clarified. This was a case-control study design with three case groups: 53 women with breast cancer and family history; 33 women with sporadic breast cancer; 175 women with no breast cancer but with family history. The control group included 120 women with no breast cancer and no family history. The PCR-RFLP method was used to analyze the XRCC1-Arg399Gln, XPD-Lys751Gln, XRCC3 Thr241Met, and RAD51-G135C polymorphisms. No statistically significant differences were found between the case groups and the control group for any of the polymorphisms analyzed, and also between the breast cancer and family history group and the sporadic breast cancer group. Sample sizes of women with breast cancer, whether familial or sporadic, were insufficient to show any small true differences between the groups, but we have to consider that currently there is no clear consensus with respect to the association of these polymorphisms with breast cancer risk. Considering the data available, it can be conjectured that if there is any risk association between these single-nucleotide polymorphisms and breast cancer, this risk will probably be minimal. The greater the risk associated with cancer, the smaller the sample size required to demonstrate this association, and the data of different studies are usually, therefore, more concordant. PMID- 16475126 TI - A genomic-scale search for regulatory binding sites in the integration host factor regulon of Escherichia coli K12. AB - We examined general aspects of the DNA-protein interaction between the integration host factor (IHF) global regulator and its regulatory binding sites in the Escherichia coli K12 genome. Two models were developed with distinct weight matrices for the regulatory binding sites recognized by IHF. Using these matrices we performed a genome scale scan and built a set of computationally predicted binding sites for each of the models. The sites found by the model associated with repetitive sequences had a higher score in the sequence to matrix alignment. They were also more rare than the other sites. The sites not associated with repeats rapidly tended to become undistinguishable from the background as statistical stringency was relaxed. We compared our results to the known sites documented in RegulonDB and found new members of the IHF Regulon. The two models exhibit clearly distinct affinity patterns (scores in the sequence to matrix alignments and in the number of regulatory sites), as we vary the stringency of the statistical confidence parameters. We suggest that these differences may play an important role in the dynamics of the network. We concluded that IHF may regulate two genes encoding ATP-dependent RNA helicases. This interaction is not described in RegulonDB, even as a computational prediction. IHF may also regulate RNA modification processes. PMID- 16475127 TI - Analysis of differential selective forces acting on the coat protein (P3) of the plant virus family Luteoviridae. AB - The coat protein (CP) of the family Luteoviridae is directly associated with the success of infection. It participates in various steps of the virus life cycle, such as virion assembly, stability, systemic infection, and transmission. Despite its importance, extensive studies on the molecular evolution of this protein are lacking. In the present study, we investigate the action of differential selective forces on the CP coding region using maximum likelihood methods. We found that the protein is subjected to heterogeneous selective pressures and some sites may be evolving near neutrality. Based on the proposed 3-D model of the CP S-domain, we showed that nearly neutral sites are predominantly located in the region of the protein that faces the interior of the capsid, in close contact with the viral RNA, while highly conserved sites are mainly part of beta-strands, in the protein's major framework. PMID- 16475128 TI - Differential molecular response of the transcripts B2A2 and B3A2 to imatinib mesylate in chronic myeloid leukemia. AB - Chronic myeloid leukemia (CML) originates from the hematopoietic stem cell and is characterized by the reciprocal translocation t(9;22)(q34;q11), which results in the BCR-ABL fusion gene on chromosome 22q-, also known as the Philadelphia chromosome. This chimeric gene codes for a cytoplasmic protein with constitutive tyrosine-kinase activity, responsible for cellular transformation and leukemogenesis in CML. The aim of this observational cohort study was to discriminate and quantify BCR-ABL transcripts in the peripheral blood of patients with CML who were treated with imatinib mesylate (Glivec, Novartis). Twenty-two patients were followed for six months during treatment. Quantitative real time polymerase chain reaction was performed before treatment and after 3 and 6 months from treatment initiation. As compared with the third month, there was a significant decrease in BCR-ABL expression in the sixth month of treatment (P = 0.0002). At the sixth month, there was a significant difference in the levels of the two major transcripts of BCR-ABL, B2A2 and B3A2 (P = 0.0347), indicating that B2A2 may be more sensitive to imatinib. The results of our study indicate that imatinib is able to modify the natural history of CML, and raise the hypothesis that patients who express the B2A2 transcript may have a better prognosis. PMID- 16475129 TI - Isolation of transfected fibroblast clones for use in nuclear transfer and transgene detection in cattle embryos. AB - Transgenesis in cattle has provided numerous opportunities for livestock production. The development of nuclear transfer (NT) technology has improved the production of transgenic livestock. However, the isolation of pure colonies from a single transfection event remains laborious and can be a constraint in the production of transgenic livestock. We used 96-well cell culture plates to isolate cell lineages obtained from a single fibroblast transfected with the pCi Neo plasmid. Since single mammalian cells do not grow well in fresh medium, we evaluated the use of conditioned medium. The neomycin phosphotransferase gene was detected in isolated colonies and NT embryos were produced from these cells. Multiplex-PCR assays were performed to detect the transfected fragment as well as autosomal satellite DNA in single NT embryos. This approach provided a reliable method for isolating transfected mammalian cells and for diagnosing the incorporation of desirable vectors in NT embryos. This method can reduce the time and cost of transgenic livestock production. PMID- 16475130 TI - Genotoxic effects of rotenone on cultured lymphocytes. AB - Rotenone is a heterocyclic compound widely used as an insecticide, acaricide and piscicide. Its toxicity is mainly caused by the inhibition of mitochondrial respiratory processes and ATP production, resulting in the generation of reactive oxygen species. Reactive oxygen species can interact with DNA, RNA and proteins, leading to cell damage, followed by death. We used the Comet assay, and we analyzed chromosome aberrations, in order to evaluate the genotoxic and clastogenic effects of rotenone on the different phases of the cell cycle. Cultured human lymphocytes were treated with 1.0, 1.5 and 2.0 microg/mL rotenone during the G1, G1/S, S (pulses of 1 and 6 h), and G2 phases of the cell cycle. Rotenone induced DNA damage and was clastogenic, but the clastogenicity was detected only with treatments conducted during the G1/S and S phases of the cell cycle. Rotenone also induced endoreduplication and polyploidy in treatments made during G1, while it significantly reduced the mitotic index in all phases of the cell cycle. PMID- 16475131 TI - Allele frequencies and genetic diversity in two groups of wild tufted capuchin monkeys (Cebus apella nigritus) living in an urban forest fragment. AB - There have been numerous studies genetically characterizing Old World Primates using microsatellites. However, few studies have been made of New World species and none on free-ranging Cebus apella, even though it is probably the most widely distributed species of monkey in the New World. The paucity of studies is due, in part, to the lack of polymorphisms described for this species. We studied two groups of wild tufted capuchins, Cebus apella nigritus, which inhabit Mata Santa Teresa, the Ecological Reserve of Ribeirao Preto, a 158-ha forest fragment in a semi-urban zone of Ribeirao Preto, SP, Brazil. Group 1 had about 60 animals, 35 of which were sampled, and group 2 had about 40 animals, 20 of which were sampled. These group sizes are much larger than the published reports of 6-30 for this species, despite, or perhaps due to the isolation and the size of the forest fragment. Allele PEPC59*1 was the most frequent of all alleles at all loci in both groups (55.7 and 55%), allele PEPC8*1 was the most common allele in group 2 (46.9%) and PEPC8*4 in group 1 (41.1%), allele PEPC3*2 was the most common in group 1 (35.7%) and allele PEPC3*4 in group 2 (31.6%). The genetic diversity, considering each locus in each group, varied from 61.9% at locus PEPC59 to 78.6% at locus PEPC3, both in group 1. The mean genetic diversity (H(S)), considering both groups for all of the loci, was 71.1%. The inter-group diversity (F(ST)) was 1.9%, indicating that these groups belong to the same population. These groups apparently have a high genetic diversity, despite their isolation in a limited forest fragment, although more data are needed to adequately characterize this population. PMID- 16475132 TI - [A retrospective study comparing two formulations of valproic acid]. AB - INTRODUCTION: Valproic acid is available in two different presentations, Depakine and Depakine Crono. Equivalences in plasmatic levels, efficacy and adverse effects between these two formulations are not fully established. AIM: To compare plasmatic levels, efficacy and plasmatic levels between Depakine and Depakine Crono. PATIENTS AND METHODS: We studied a retrospective series of 238 patients in treatment with Depakine or Depakine Crono and compared plasmatic levels, efficacy and adverse effects between both formulations. RESULTS: A total of 173 patients had taken one formulation and 54 both. Plasmatic levels were similar in both groups. There were more significantly more patients free of seizures among the Depakine Crono group. Significantly better outcomes were seen with Depakine Crono with respect to secondary effects. Adverse effects are significantly less frequent in men than in women. The higher the dose, the higher the incidence of adverse events. CONCLUSIONS: Depakine Crono seems to obtain similar plasmatic levels, is at least as efficacious and is associated with less adverse events that Depakine. PMID- 16475134 TI - [Familial impact of children with the combined subtype of attention deficit hyperactivity disorder: the effects of associated behavioural disorders]. AB - AIMS: To determine the impact that children with ADHD-C (attention deficit hyperactivity disorder, combined subtype) have on their family by analysing their parents' perceptions, and to examine whether the presence of associated behavioural disorders affect that impact. SUBJECTS AND METHODS: Participants in the study included one group made up of the parents of 27 children with ADHD-C and another group consisting of the parents of 27 children without ADHD. The parents in the ADHD-C group were divided into two subgroups according to whether or not their children had an oppositional defiant disorder and/or a co-morbid conduct disorder (10 and 17 families, respectively). RESULTS AND CONCLUSIONS: The results of the analyses performed showed significant differences between the perceptions of the parents of the control children and those of the parents of children with ADHD-C in the following categories: feelings and attitudes, social life, matrimonial relationship, day-to-day relationships with peers and siblings, stress and difficulty in living with their child. Comparisons between the two subgroups of children with ADHD-C did not reveal any significant differences in any of the categories that were analysed, and showed ADHD-C to be the fundamental factor underlying the problems in the family context. Some items, however, suggested that the problem is more serious in the subtype with associated behavioural disorders. PMID- 16475133 TI - [Cognitive slowing in asymptomatic individuals who are seropositive for human immunodeficiency virus type 1]. AB - INTRODUCTION: Reaction time (RT) is thought to be the most suitable measure to detect cognitive deficits in neurologically asymptomatic human immunodeficiency virus type I (HIV-1) infected individual since cognitive slowing is the earliest signal of cognitive-motor disorder related to HIV-1 infection. There is evidence suggesting that the greater the degree of central processing demands required by a task, the more likely that it will be sensitive to the effect of HIV-1 infection. Such statement suggests that the RT deficits exhibited by HIV-1 infected individuals at initial stages could be caused by the slowing of central information processing mechanisms. AIM: To assess the relationships between demands of central information processing and RT in HIV-1 seropositive individuals. SUBJECTS AND METHODS: 50 neurologically asymptomatic HIV-1 individuals were compared with 34 seronegative controls on four discriminative RT tasks of different levels of central processing demands except by the motor response requirements. RESULTS: Seropositive group was slower in RT and performed worse on the higher demanding task. On the lesser demanding tasks no differences in RT nor in accuracy were observed. For the task demanding sensory coding efforts seropositive individual were slower but achieved the same level of accuracy. CONCLUSIONS: Even when these results point to that RT slowing in HIV-1 asymptomatic individuals emerged with the increase in cognitive demands, the fact that RT slowing without accuracy declining can also appear in some tasks demanding sensory processing, preclude ruling out a peripheral deficit as the locus of the RT slowing in these subjects. PMID- 16475135 TI - [Pediatric Guillain-Barre syndrome. Experience in the Neuropediatrics Unit of a Portuguese hospital]. AB - INTRODUCTION: Guillain-Barre syndrome (GBS) is an acute inflammatory ascending polyradiculoneuropathy. Autoimmune mechanisms play a role in the demyelinating process. Clinically, progressive symmetric loss of motor strength, areflexia, sensitive and autonomic manifestations are observed. Albuminocytological dissociation and electrophysiological signs of demyelination are frequently found. It is the most common cause of acute flaccid paralysis in children. PATIENTS AND METHODS: Retrospective review of all children with GBS admitted to Garcia de Orta Hospital in a 10 year period (1994-2003). RESULTS: 17 children (18 months to 14 years) were admitted during this period. Respiratory or gastrointestinal prodrome was identified in 15 children, with positive serologic studies in six. The main clinical manifestations were disturbances of gait, progressive muscular weakness, sensitive manifestations (pain, paresthesias) and osteotendinous areflexia. 13 children had albuminocytological dissociation and acute demyelinating neuropathy was identified in 12.64% of children were treated with immunoglobulins (2 g/kg). Clinical evolution was favourable in 16 cases, with a death secondary to autonomic dysfunction. CONCLUSIONS: Clinical presentation may be unspecific, particularly in young patients, with pain as a primary complaint, preceding muscular weakness and areflexia. Increased cerebrospinal protein and abnormal electrodiagnostic studies may be absent in the early course of GBS. Immunoglobulin therapy was efficacious and well-tolerated. PMID- 16475136 TI - [Early onset Neurobehcet's disease]. AB - INTRODUCTION: Behcet's disease is difficult to diagnose in infancy due to the absence of early clinicopathological pathognomonic symptoms. There have been reports of patients in whom the onset of the disease occurred during the first months of their lives. CASE REPORT: We describe here the case of a 12-year-old female from Pakistan whose first symptoms appeared at the age of 6 months in the form of fever and skin lesions. The first neurological symptoms appeared at the age of 5 years in the form of ophthalmoplegia, followed by ataxia and dysmetry. Magnetic resonance imaging of the head was a valuable aid in the study of the vascular alterations. She also presented abdominal pains, arthromyalgia, ulcers in the mouth and severely delayed weight gain. Yet, we were finally guided towards a diagnosis by the appearance of ulcers on the genitals, which do not usually occur until puberty, as well as the findings from the skin biopsy study. The serological study revealed the presence of antigen B08. Establishing therapy with methylprednisolone and later with azathioprine brought about the practically complete remission of the clinical picture. CONCLUSIONS: Behcet's disease, which has been infrequent in our country up till now, is a condition to be taken into account, especially in young female patients from countries with a high incidence rate of oral and genital ulcers, accompanied by neurological symptoms. Knowledge of this syndrome and performing appropriate complementary studies are essential to be able to reach an early diagnosis and for establishing suitable treatment. PMID- 16475137 TI - [Plasticity of the central nervous system and treatment strategies for sensory and motor reprogram: comparison between two cases of cerebrovascular disease in the middle cerebral artery territory]. AB - INTRODUCTION: Injury to the central nervous system causes a series of symptoms, which lead to a process of cerebral plasticity. Proprioceptive and exteroceptive information processing from the outside to the brain, influence the development of neuroplasticity, which contributes to either the recovery or maintenance of sensory-motor alterations after a stroke. CASE REPORTS: Two patients with cerebrovascular accident in the left middle cerebral artery territory, one of them ischemic and the other hemorrhagic, whose sensory and motor therapy applied show big differences. CONCLUSIONS: Treatment strategies for functional rehabilitation following a cerebrovascular accident, must be established in relation to current hypotheses on cerebral neuroplasticity and central nervous system pathways, which shows less damage or better sensory motor response. The assessment of neurological signs and symptoms, can indicate which tracts show more or less damage and which ones show greater response to motor recovery treatment. Nevertheless, functional neuroimaging diagnosis provides us with the data in a more reliable way. Even so, among patients that show similar lesion zones, according to diagnosis of computerized tomography or magnetic resonance imaging, there exists a significant clinical variability among different cases, in which case we confirm, in reference to physical treatment, that there are no patients alike, or identical patients even if they have the same clinical diagnosis of the disease. PMID- 16475138 TI - [The neurochemistry of epilepsy, inhibitory neurotransmission and experimental models: new perspectives]. AB - AIMS: The biological mechanisms of epilepsy allow pathophysiological patterns to be established which are essential for the selection of new therapeutic targets. The identification of these mechanisms also provides us with knowledge about the dynamics of neuronal arrangement, synaptogenesis, synaptic transmission and the receptors involved, and even the development of the brain. DEVELOPMENT: Recent advances in neurobiology regarding the GABAergic system point to it as playing a leading role in the pathophysiology of epilepsy. We evaluate the different functional formats of the ionotropic (GABAA) and metabotropic (GABAB) gamma aminobutyric (GABA) receptors. Although the main function posited is inhibitory, owing to the variability of their location, subunits and neuronal physiology/maturation they can even end up expressing excitatory functions. We discuss the anomalies in the GABAergic system identified in animal models with epilepsy and in brain tissue samples from patients submitted to surgery due to epilepsy. The mechanism inhibiting the activation of GABA receptors is performed by hyperpolarisation achieved by entry of Cl- into the neuron--a process mediated by the cotransporter KCC2, typically expressed in the neuron. Mutations in the KCC2 gene produce mice that are susceptible to seizures. In some animal models it has been found that loop diuretics (furosemide) suppress seizures. Mutations in genes that code for ion channels have been identified in numerous epileptic syndromes and this pushes epilepsy ever further inside the broad group of disorders known as channelopathies. The origin could be polygenetic in many cases. CONCLUSIONS: The GABAergic system seems to situate itself as the main system implicated in the pathophysiology of epilepsy, although conditions that have been considered to be idiopathic up till now could have a polygenic nature. PMID- 16475139 TI - [Post-stroke depression (I). Epidemiology, diagnostic criteria and risk factors]. AB - AIM: Post-stroke depression (PSD) is the most frequent neuropsychiatric condition after a stroke. We review the diagnostic criteria, differential diagnosis, type of scales applied, the epidemiological studies and associated risk factors in PSD. DEVELOPMENT: PSD may be considered early when appear in the first three months after a stroke or late. We reviewed PSD case series, Stroke Unit or rehabilitation Unit studies, and community studies. Frequency of PSD has been estimated between 18 and 60%. Several methodological problems regarding inclusion criteria (acute vs chronic patients, exclusion of demented and aphasic patients), type of scales used (self applied scales, DSM-IV criteria interview, Hamilton, etc.) limit direct comparison between studies. Symptoms of PSD appear in three areas: cognition, affective and somatic. Relation between stroke location in left frontal region and left basal ganglia has not been demonstrated in further epidemiological studies. CONCLUSION: Post-stroke depression is a highly prevalent, potentially treatable and infra-diagnosed condition. Disparity in inclusion criteria, study timing and types of scales are the main bias of the PSD epidemiological studies. PMID- 16475140 TI - [Cerebral vasculitis]. AB - INTRODUCTION: Cerebral vasculitis embraces a wide range of conditions that are characterised by involvement of the vessels of the central nervous system (CNS) due to inflammation of their walls, which in turn leads to occlusion or the formation of aneurysms with the ensuing ischaemic-haemorrhagic disorders this produces. DEVELOPMENT: In cases of cerebral vasculitis perhaps only the vessels of the CNS (isolated angiitis of the CNS) are involved, or they may be just another of the affected territories to be found in primary or secondary systemic angiitis (due to infection, collagen diseases, drugs, tumours). Neurological symptoms and lab tests are usually rather unspecific. The latest neuroimaging techniques are more useful, and the gold standard among them is digital angiography, although its sensitivity and specificity are limited. Brain tissue biopsy allows for confirmation of the diagnosis and is the gold standard for the diagnosis of isolated angiitis of CNS. A large group of conditions (which may be metabolic, demyelinating, vascular, infectious, and other peripheral vascular diseases) have similar clinical and imaging features, which makes it necessary to consider the differential diagnosis. CONCLUSIONS: Involvement of the nervous system casts a shadow over the prognosis in most cases of vasculitis and can be severe, as in isolated vasculitis of large vessels or in Takayasu's disease, or more benign, as in isolated vasculitis of small vessels and in other primary vasculitis. Treatment with corticoids and immunosuppressant agents, as well as anticoagulant and/or antiaggregating therapy, must be considered in each particular case according to the clinical condition and the progression of each patient. PMID- 16475141 TI - [Migrainous auras without cephalalgia in crescendo and optico-cerebral syndrome as clinical presentation of an atrial myxoma]. PMID- 16475142 TI - [Giant cell arteritis in association with acute multifocal stroke]. PMID- 16475143 TI - [Some interesting aspects of headaches in infancy]. PMID- 16475144 TI - [The role of paediatrics in the diagnosis and treatment of attention deficit hyperactivity disorder]. PMID- 16475146 TI - MR angiography fusion technique for treatment planning of intracranial arteriovenous malformations. AB - PURPOSE: To develop an image fusion technique using elliptical centric contrast enhanced (CE) MR angiography (MRA) and three-dimensional (3D) time-of-flight (TOF) acquisitions for radiosurgery treatment planning of arteriovenous malformations (AVMs). MATERIALS AND METHODS: CE and 3D-TOF MR angiograms with disparate in-plane fields of view (FOVs) were acquired, followed by k-space reformatting to provide equal voxel dimensions. Spatial domain addition was performed to provide a third, fused data volume. Spatial distortion was evaluated on an MRA phantom and provided slice-dependent and global distortion along the three physical dimensions of the MR scanner. In vivo validation was performed on 10 patients with intracranial AVMs prior to their conventional angiogram on the day of gamma knife radiosurgery. RESULTS: Spatial distortion in the phantom within a volume of 14 x 14 x 3.2 cm(3) was less than +/-1 mm (+/-1 standard deviation (SD)) for CE and 3D-TOF data sets. Fused data volumes were successfully generated for all 10 patients. CONCLUSION: Image fusion can be used to obtain high-resolution CE-MRA images of intracranial AVMs while keeping the fiducial markers needed for gamma knife radiosurgery planning. The spatial fidelity of these data is within the tolerance acceptable for daily quality control (QC) purposes and gamma knife treatment planning. PMID- 16475145 TI - Evaluation of diffusion-weighted imaging for the differential diagnosis of poorly contrast-enhanced and T2-prolonged bone masses: Initial experience. AB - PURPOSE: To determine whether quantitative diffusion-weighted imaging (DWI) is useful for characterizing poorly contrast-enhanced and T2-prolonged bone masses. MATERIALS AND METHODS: We studied 20 bone masses that showed high signal intensity on T2-weighted images and poor enhancement on contrast-enhanced T1 weighted images. These included eight solitary bone cysts, five fibrous dysplasias, and seven chondrosarcomas. To analyze diffusion changes we calculated the apparent diffusion coefficient (ADC) for each lesion. RESULTS: The ADC values of the two types of benign lesions and chondrosarcomas were not significantly different. However, the mean ADC value of solitary bone cysts (mean +/-SD, 2.57 +/- 0.13 x 10(-3) mm(2)/second) was significantly higher than that of fibrous dysplasias and chondrosarcomas (2.0 +/- 0.21 x 10(-3) mm(2)/second and 2.29 +/- 0.14 x 10(-3) mm(2)/second, respectively, P < 0.05). None of the lesions with ADC values lower than 2.0 x 10(-3) mm(2)/second were chondrosarcomas. CONCLUSION: Although there was some overlapping in the ADC values of chondrosarcomas, solitary bone cyst, and fibrous dysplasia, quantitative DWI may aid in the differential diagnosis of poorly contrast-enhanced and T2-prolonged bone masses. PMID- 16475147 TI - In vivo administered reserpine increases piecemeal degranulation in rat adrenal chromaffin cells. AB - The effects of the amine-depletory agent reserpine have been evaluated by transmission electron microscopy in chromaffin cells of the rat adrenal glands. The drug has been injected intraperitoneally in the animals at a dose of 0.5 mg/kg body weight in two administrations at 24-hr interval. The observed ultrastructural changes closely reminded of piecemeal degranulation (PMD), a slow and long-lasting secretory process previously described in normal and tumor pheochromocytes. Both adrenaline- and noradrenaline-storing cells presented the following microscopic features: high granule polymorphism, due to coexistence in the same cell of normal resting granules, granules with partially mobilized components, and large empty containers; absence of granule fusion; characteristic "haloed" pattern of residual secretory contents; great amount of 30-150 nm diameter, membrane-bound, electron-dense and -lucent vesicles, free in the cytoplasm or attached to granules; and multiple vesicles budding from the granule limiting membranes. Morphometric analysis revealed that the frequency of all these microscopic parameters was found to be significantly increased in adrenal chromaffin cells from reserpinized rats in comparison to cells from control animals. These data suggest that reserpine, besides blocking the inward transport of catecholamines in chromaffin granules, might also stimulate a complex secretory reaction, which shares many common passages with bona fide PMD. PMID- 16475148 TI - Maturation of dendritic cells and T-cell responses in sentinel lymph nodes from patients with breast carcinoma. AB - BACKGROUND: Methods for identifying sentinel lymph nodes (SNs) and their clinical significance have been established. Recent advances in molecular immunology have enabled the analysis of precise immune responses. The objective of the current study was to clarify the dendritic cell (DC) maturation, T-helper type 1 (Th-1) and Th-2 responses, and regulatory T-cell responses of SNs in patients with breast carcinoma. METHODS: SNs and non-SNs were identified by radioguided and blue dye-guided methods in 70 consecutive patients with clinically lymph node negative (N0) breast carcinoma. Lymphocytes were collected from SNs and non-SNs and were subjected to flow cytometric analysis (FCM) using antibodies of CD83 fluorescein isothiocyanate (FITC), CD80-phycoerythrin (PE), CD86-PE, CD40-PE, human leukemic D-related antigen (HLA-DR)-FITC, CD4-FITC, and CD25-PE. Total RNA was extracted from SNs and non-SNs, and the expression of CD83, interleukin 12p40 (IL-12p40), interferon gamma (IFN-gamma), IL-4, IL-10, and Foxp3 was evaluated by using quantitative real-time reverse transcriptase-polymerase chain reaction (RT PCR) analysis. The immunologic status of SNs was analyzed further with regard to micrometastases, which were identified as negative microscopically but positive according to an RT-PCR analysis that was specific for mammaglobin. RESULTS: SNs were detectable in 70 of 71 consecutive patients (98.6%) with clinically N0 breast carcinoma. Fourteen of 70 patients (20.0%) had positive metastasis in SNs. When SNs were compared with non-SNs in 56 metastasis-negative patients, FCM revealed that HLA-DR-positive, CD80-positive, CD86-positive, and CD40-positive cell populations were decreased significantly in SNs. RT-PCR analysis demonstrated that, among 44 patients with metastasis-negative SNs, the expression levels of CD83 and IFN-gamma mRNA were significantly lower in SNs compared with non-SNs. Immunologic parameters also were compared between 44 metastasis-negative SNs and 14 metastasis-positive SNs. The metastasis-positive SNs demonstrated significantly higher expression of CD83, IL-12p40, IFN-gamma, IL-10, and Foxp3 mRNA than the metastasis-negative SNs. Correction of micrometastasis detected by mammaglobin enhanced these differences consistently. CONCLUSIONS: In patients with breast carcinoma, cellular immune responses, from DC maturation to Th-1 responses, appeared to be less active in SNs compared with non-SNs before metastasis developed. Once metastasis was established in SNs, DC maturation was triggered and was followed by the up-regulation of Th-1 responses, which may reflect antigen-specific immune responses in SNs. Unlike DC maturation and Th-1 responses after metastasis in SNs, up-regulation of Th-2 and regulatory T-cell responses developed in parallel. PMID- 16475149 TI - Characterization of bone marrow T cells in monoclonal gammopathy of undetermined significance, multiple myeloma, and plasma cell leukemia demonstrates increased infiltration by cytotoxic/Th1 T cells demonstrating a squed TCR-Vbeta repertoire. AB - BACKGROUND: The majority of studies published to date regarding the role of the bone marrow (BM) microenvironment in the pathogenesis of monoclonal gammopathies (MG) have focused on the interaction between stroma cells and plasma cells, whereas information concerning the lymphocytes infiltrating the tumor microenvironment is scanty. METHODS: The authors measured the distribution, TCR Vbeta repertoire, immunophenotype, and functional characteristics of different subsets of BM T lymphocytes from 61 nontreated patients with MG (30 patients with MG of undetermined significance [MGUS], 27 patients with multiple myeloma [MM], and 4 patients with plasma cell leukemia [PCL]). RESULTS: The authors found a significantly increased rate of BM infiltration by T cells in all patient groups, at the expense of CD4+CD8- and CD4-CD8- T lymphocytes and both CD4+CD28- and CD8+CD28- cytotoxic/effector T cell subsets, and associated with TCR-Vbeta expansions in both CD4+ and CD8+ BM T cells in the majority of patients with MGUS, MM, and PCL. Moreover, the percentage of T cells secreting interferon (IFN) gamma was found to be increased (P < or = 0.05) both in CD4+ and CD8+ T cells in MGUS and MM patients, and a higher plasma concentration of IFN-gamma was found in patients with MM. It is interesting to note that a positive correlation was noted between the proportion of CD28- and both the percentage of IFN-gamma-secreting cells and the proportion of expanded TCR-Vbeta lymphocytes within the total BM CD4+ T cells. CONCLUSIONS: The results of the current study demonstrated an increased infiltration of BM by T cells associated with frequent TCR-Vbeta expansions and a more prominent cytotoxic/Th1 phenotype in all the patient groups studied. PMID- 16475150 TI - Clinical outcomes of single-fraction stereotactic radiation therapy of lung tumors. AB - BACKGROUND: The objective of the current study was to investigate the effects and the morbidities of single-fraction stereotactic radiation therapy (SRT) for lung tumors. METHODS: A Microtron device was modified to deliver stereotactic irradiation under respiratory gating. Between August 1998 and December 2004, 59 malignant lung tumors (11 primary tumors, 48 metastases) that measured < 40 mm in greatest dimension were treated by single-fraction SRT. Nine tumors received a minimal dose of < 30 grays (Gy), and 50 tumors received a minimal dose of > or = 30 Gy. The macroscopic target volume ranged from 1 cc to 19 cc (mean, 5 cc). RESULTS: The 1-year and 2-year local progression-free rates (LPFRs) were 93% and 78%, respectively. The overall survival rate was 76.5% at 1 year and 41% at 2 years. Local regrowth of the irradiated tumor was a direct cause of death in two patients. Only the minimal radiation dose to the reference target volume tended to have an influence on the LPFR (P = 0.068). The 2-year LPFRs for patients who received irradiation doses of > or = 30 Gy and < 30 Gy were 83% and 52%, respectively. With regard to morbidities, Grade 3 respiratory symptoms (according to the Radiation Therapy Oncology Group/European Organization for Research and Treatment of Cancer late radiation morbidity scoring scheme) were noted in one patient. CONCLUSIONS: The results from the current study suggested that single fraction SRT was tolerable and was capable of attaining excellent local control in patients who had malignant lung tumors that measured < 4 cm in greatest dimension. PMID- 16475151 TI - Survival analysis after resection of metastatic disease followed by peptide vaccines in patients with Stage IV melanoma. AB - BACKGROUND: Metastatic melanoma carries a poor prognosis, with a median survival of 7-9 months. Surgical resection of metastatic disease has been advocated to improve survival. Immunotherapy after metastasectomy may further improve the outcome for high-risk resected disease. METHODS: Charts from patients treated on institutional vaccine trials were analyzed. Patients with American Joint Committee on Cancer (AJCC) Stage IV melanoma who underwent surgical resection of metastatic sites followed by treatment on a peptide vaccine trial were eligible for this study. Survival was calculated from the date of enrollment on the clinical trial. RESULTS: Forty-one patients met inclusion criteria. The median age was 56.5 years, with approximately equal numbers of men and women. The ECOG performance status was 0 in all patients. Approximately 46% of patients underwent resection of visceral metastases before vaccine. The median follow-up was 5.6 years. The median overall survival was 3.8 years. CONCLUSIONS: In selected patients with AJCC Stage IV melanoma, resection of metastatic disease followed by vaccine therapy can result in long-term survival. PMID- 16475152 TI - Phase I/II study of thalidomide in combination with interleukin-2 in patients with metastatic renal cell carcinoma. AB - BACKGROUND: The purpose of the study was to determine, in a Phase I/II study, the efficacy and safety profile of thalidomide with interleukin-2 (IL-2) in patients with metastatic renal cell carcinoma (MRCC). METHODS: Fifteen patients (8 of whom were previously treated) enrolled in Phase I were treated with escalating doses of oral thalidomide (200-600 mg) and a fixed dose of IL-2 (7 mIU/m(2)) by subcutaneous injection. A course was 6 weeks, with the exception of Course 1, which was 7 weeks. Thirty-seven Phase II patients who had not received prior chemotherapy or immunotherapy for renal cell carcinoma (RCC) received an initial thalidomide dose of 200 mg at Week 0, which was escalated to 400 mg after 48 hours. Subcutaneous IL-2 was administered at the same fixed daily dose used in Phase I. RESULTS: Fifty-one of 52 Phase I/II patients were evaluable. Twenty seven patients (52%) experienced disease control, including 4 (8%) complete responses, 15 (29%) partial responses, and 8 (15%) cases of stable disease. Disease progression was observed in 24 patients (47%). Survival in the 2 phases ranged from 4 weeks to 45.2+ months. At the time of last follow-up, 2 of 51 patients (4%) remained on maintenance thalidomide therapy and continue to be followed. Three of the 51 patients with CRs (6%) ceased thalidomide therapy at 23 25 months and have maintained their responses to date. One complete responder was lost to follow-up. As of January 2005, 14 of 51 patients (27%) remained alive. Toxicities were mild to moderate, including Grade 1 to 2 somnolence, constipation, neuropathy, rash, flu-like symptoms, fluid retention, hypotension, and hypothyroidism (according to version 2.0 of National Cancer Institute Common Toxicity Criteria). In addition, two patients experienced deep venous thrombosis. CONCLUSIONS: Thalidomide in combination with IL-2 is tolerable and can produce durable, active responses in patients with MRCC. To evaluate the merits of thalidomide as a valuable agent against MRCC and to more fully determine the efficacy of thalidomide/IL-2 combination therapy, the scrutiny of Phase III testing is required. Further development of thalidomide/IL-2 combination therapy will be the focus of this group. PMID- 16475154 TI - Cognitive function in multiple system atrophy of the cerebellar type. AB - Dementia represents an exclusion criterion for the diagnosis of multiple system atrophy (MSA), but there have been reports of fronto-executive dysfunction in patients with MSA of the striatonigral type (MSA-P). To study the cognitive profile of MSA, 20 patients with MSA of the cerebellar type (MSA-C) were subjected to an extensive neuropsychological test battery comprising tests for IQ, attention, verbal and visuospatial memory, as well as executive function. There was evidence for impaired verbal memory and verbal fluency. Test performance was not related to the severity of motor disability. Regarding the similar cognitive syndrome of MSA-P, the otherwise subclinical problems in MSA-C result from subcortical rather than from cerebellar dysfunction. PMID- 16475153 TI - Clinical and radiologic predictors of invasive pulmonary aspergillosis in cancer patients: should the European Organization for Research and Treatment of Cancer/Mycosis Study Group (EORTC/MSG) criteria be revised? AB - BACKGROUND: Invasive pulmonary aspergillosis (IPA) is a life-threatening infection in immunocompromised patients. The outcome of such infections depends on early diagnosis and prompt initiation of therapy. The objective of the current study was to determine the significant predictive factors that characterize IPA in patients with cancer. METHODS: The clinical characteristics and computed tomography (CT) findings for 47 cases with IPA were retrospectively reviewed and compared with 49 controls (patients diagnosed on autopsy with nonfungal pneumonias). Data from all 96 patients were modeled using multivariate logistic regression. Subgroups of patients with common characteristics and outcomes were identified. RESULTS: Leukemia, neutropenia, cavitation, and nodular lesions occurred significantly more often among cases than controls (P = 0.04, 0.004, 0.04, and 0.02, respectively). A quantitative scoring system was developed that could be used to identify patients as being at low, medium, and high risk for IPA. CONCLUSIONS: IPA should be highly suspected in leukemia patients with profound neutropenia, pleuritic chest pain, and cavitary or nodular lesions detected on CT scan. These predictive factors can be used to indicate when early prophylactic and therapeutic antifungal interventions should be initiated. PMID- 16475155 TI - Conformational features of truncated hepatitis C virus core protein in virus-like particles. AB - HCVc 120 is a truncated protein from the hepatitis C virus (HCV) core protein that interacts with itself to form nucleocapsid-like particles. We present here the infrared and Raman spectra of oligomeric HCVc 120 protein in order to obtain insights into its secondary structure as well as the environment surrounding some protein side chains. When compared with its monomer form, oligomeric HCVc 120 protein shows an increase in beta-sheet structure. Tryptophan residues have been found to be solvent exposed in the oligomeric form, and they likely do not significantly participate in the protein assembly. However, the beta-sheet content in oligomeric HCVc 120 protein suggests that this structural motif cannot be excluded in nucleocapsid formation, as shown recently in other viruses. PMID- 16475156 TI - The trans-cis isomerization reaction dynamics in sensory rhodopsin II by femtosecond time-resolved midinfrared spectroscopy: chromophore and protein dynamics. AB - Transient infrared (IR) vibrational spectroscopy at subpicosecond time resolution on sensory rhodopsin II from Natronomonas pharaonis, NpSRII, has been performed for the first time. The experiments yield three time constants for the description of the primary photoinduced reaction dynamics, i.e. 0.5, 3.7-4.4, and 11 ps. The data are consistent with a sequential reaction scheme, with the isomerization taking place within 0.5 ps, succeeded by an electronic ground state relaxation. The 11 ps component, observed at 1550 and 1530 cm(-1), is attributed to dynamics of protein vibrational bands, possibly amide II bands of the protein backbone, perturbed by the ultrafast retinal photoisomerization. Similar observations, yet not as strongly expressed, have been made earlier in bacteriorhodopsin and halorhodopsin. PMID- 16475157 TI - Effects of rare earth ions on the conformational stability of anticoagulation factor II from Agkistrodon acutus venom probed by fluorescent spectroscopy. AB - Anticoagulation factor II (ACF II) isolated from the venom of Agkistrodon acutus is an activated coagulation factor X-binding protein in a Ca(2+)-dependent fashion with marked anticoagulant activity. The equilibrium unfolding of rare earth ions (RE(3+))-reconstituted ACF II in guanidine hydrochloride (GdnHCl) solution was studied by fluorescence. The GdnHCl-induced unfolding of RE(3+) (Nd(3+), Sm(3+), Eu(3+), Gd(3+))-reconstituted ACF II follows a three-state transition with a stable intermediate state. Substitutions of the RE(3+) ions for Ca(2+) in ACF II decrease the conformational stability of its native state but markedly increase the conformational stability of its intermediate state. The free energy change of RE(3+)-ACF II from the intermediate state to denatured state linearly increases with the increase of ionic potentials of bound metal ions (Ca(2+), Nd(3+), Sm(3+), Eu(3+), and Gd(3+)). The refolding of ACF II from the unfolded state to the intermediate state can be induced merely by adding 10 microM RE(3+) ions without changing the concentration of the denaturant. The kinetic results of the RE(3+)-induced refolding provide evidence indicating that the intermediate state of RE(3+)-ACF II consists of at least two refolding phases and that the refolding rate constant values of the faster phase decrease with the increase of the difference between the radii of Ca(2+) and RE(3+), but the refolding rate constant values of the slower phase are similar to each other. The results of this study indicate that the size of metal ion is the major factor responsible for the metal ion-induced conformational stabilization of the native ACF II, while the metal ionic potential plays a predominant role in stabilizing the conformation for the intermediate state. PMID- 16475158 TI - Fourier transform infrared spectroscopic study on the Ca2+ -bound coordination structures of synthetic peptide analogues of the calcium-binding site III of troponin C. AB - The coordination structures of Ca(2+) ion bound to synthetic peptide analogues of the calcium-binding site III of rabbit skeletal muscle troponin C (TnC) were investigated by Fourier transform infrared (FTIR) spectroscopy. The region of the COO(-) antisymmetric stretching vibration provides information about the coordination modes of a COO(-) group to a metal ion. The 34-residue peptide corresponding to the EF hand motif (helix-loop-helix) showed a band at 1552 cm( 1) in the Ca(2+)-loaded state, indicating that the side-chain COO(-) group of Glu at the 12th position serves as a ligand for Ca(2+) in the bidentate coordination mode. On the other hand, the 13-residue peptide (Ac-DRDADGYIDAEEL-NH(2)) containing the Ca(2+)-binding site III (DRDADGYIDAEE) did not show such spectral patterns in the Ca(2+)-loaded state, meaning that shorter synthetic peptide corresponding to the site III has less or no affinity for Ca(2+). It was found that the 17-residue peptide (Ac-DRDADGYIDAEELAEIF-NH(2)) is the minimum peptide necessary for the interaction of side-chain COO(-)of Glu at the 12th position with Ca(2+) in the bidentate coordination mode. We discuss the relationship between the amino acid length of synthetic peptide analogues and the formation of Ca(2+)-bound coordination structure. PMID- 16475159 TI - Diagnostic accuracy of skin-prick testing in young children with asthma. PMID- 16475160 TI - Role of soluble ceruloplasmin in iron uptake by midbrain and hippocampus neurons. AB - Ceruloplasmin (CP) is essential for brain iron homeostasis. However, its precise function in brain iron transport has not been definitely determined. In this study, we investigated the effects of soluble CP on iron influx and efflux in primary neuronal culture from the midbrain (the substantia nigra and striatum) and the hippocampus. Our data showed that low concentrations of CP (2, 4, 8 microg/ml) can promote iron influx into iron-deficient neurons, but not the neurons with normal iron status. The same concentrations of CP had no effect on iron efflux from iron-sufficient and normal-iron neurons. Contrary to our expectation, we did not find any regional difference in the effects of CP on iron influx as well as efflux in neurons. The changes in quenching (iron influx) and also dequenching (iron efflux) of intracellular fluorescence, induced by the addition of CP with iron, in the midbrain neurons were no different from those in the hippocampus neurons. The data showed that soluble CP has a role in iron uptake by iron-deficient brain neurons under our experimental conditions. The physiological significance of the results forms the focus for future work. PMID- 16475161 TI - Role of protein phosphatase 2A in the regulation of endothelial cell cytoskeleton structure. AB - Our recently published data suggested the involvement of protein phosphatase 2A (PP2A) in endothelial cell (EC) barrier regulation (Tar et al. [2004] J Cell Biochem 92:534-546). In order to further elucidate the role of PP2A in the regulation of EC cytoskeleton and permeability, PP2A catalytic (PP2Ac) and A regulatory (PP2Aa) subunits were cloned and human pulmonary arterial EC (HPAEC) were transfected with PP2A mammalian expression constructs or infected with PP2A recombinant adenoviruses. Immunostaining of PP2Ac or of PP2Aa + c overexpressing HPAEC indicated actin cytoskeleton rearrangement. PP2A overexpression hindered or at least dramatically reduced thrombin- or nocodazole-induced F-actin stress fiber formation and microtubule (MT) dissolution. Accordingly, it also attenuated thrombin- or nocodazole-induced decrease in transendothelial electrical resistance indicative of barrier protection. Inhibition of PP2A by okadaic acid abolished its effect on agonist-induced changes in EC cytoskeleton; this indicates a critical role of PP2A activity in EC cytoskeletal maintenance. The overexpression of PP2A significantly attenuated thrombin- or nocodazole-induced phosphorylation of HSP27 and tau, two cytoskeletal proteins, which potentially could be involved in agonist-induced cytoskeletal rearrangement and in the increase of permeability. PP2A-mediated dephosphorylation of HSP27 and tau correlated with PP2A-induced preservation of EC cytoskeleton and barrier maintenance. Collectively, our observations clearly demonstrate the crucial role of PP2A in EC barrier protection. PMID- 16475162 TI - Identification of nuclear localization signal that governs nuclear import of BRD7 and its essential roles in inhibiting cell cycle progression. AB - BRD7, a novel bromodomain gene, is identified to be associated with nasopharyngeal carcinoma (NPC). Decreased or loss of expression of BRD7 was detected in NPC biopsies and cell lines. Overexpression of BRD7 could inhibit NPC cell growth and arrest cells in cell cycle by transcriptionally regulating some important molecules involved in ras/MEK/ERK and Rb/E2F pathway, and downregulate the promoter activity of E2F3. In the present study, the subcellular localization of BRD7 was investigated. It was found that BRD7 was mainly localized in nucleus without distinct cell-specific difference between COS7 and HNE1. Furthermore, a functional nuclear localization signal (NLS) sequence ranging from amino acid 65 to 96 was identified and characterized. The NLS is composed of a cluster of four bipartite nuclear targeting sequences, which are tightly linked and extremely overlapped. We found that whether the entire NLS or the four bipartite nuclear targeting sequences could respectively determine the nuclear import of green fluorescent protein (GFP). The most important is that NLS-deleted BRD7 shifted the nuclear localization to be mostly in cytoplasm, and failed or reduced to negatively regulate the expression of cell cycle related molecules, cyclin D1 and E2F3, and cell cycle progression from G1 to S phase. In conclusion, NLS is an essential motif affecting BRD7 nuclear distribution, and the nuclear localization of BRD7 is critical for the expression of cell cycle related molecules and cell biological function. PMID- 16475163 TI - Coronary sinus is dilated and outwardly displaced in patients with mitral regurgitation: quantitative angiographic analysis. AB - OBJECTIVES: Our aim was to define the anatomy of the coronary sinus (CS) by venography. These measurements are essential in the selection of physical characteristics of percutaneous annuloplasty devices for mitral regurgitation (MR). BACKGROUND: Clinical trials of percutaneous annuloplasty of the CS for MR are now underway. Although the CS is in close proximity to the mitral annulus, there is as yet no published quantitative data defining the magnitude of change in CS dimensions in MR, and how these changes might reflect the magnitude of MR. METHODS: We studied 57 patients (27 patients with MR and 30 patients with no MR) who were referred for cardiac resynchronization therapy and underwent CS venography. Echocardiography was used to assess the degree of MR, cardiac dimensions, and right heart filling pressures. The diameter of the ostial and proximal CS and perimeter of the CS-great cardiac vein (GCV) were assessed by quantitative coronary analysis. RESULTS: Patients with MR had a larger ostial CS diameter (19.4+/-3.9 mm vs. 16.9+/-4.6 mm, P=0.02) and proximal CS diameter (8.2+/-1.7 mm vs. 7.4+/-2.3 mm, P=0.05) and larger CS-GCV perimeter (104.4+/-15.6 mm vs. 86.5+/-15.3 mm, P=0.005) compared with patients with no MR. The CS-GCV perimeter is positively correlated to the severity of MR (P=0.02) and pulmonary artery pressure (r=0.32, P<0.05). CONCLUSIONS: Patients with MR have a dilated and outward displacement of the CS. The CS-GCV perimeter is positively correlated with the degree of MR and pulmonary artery pressure. PMID- 16475164 TI - Implantation of oxygen enhanced, three-dimensional microporous L-PLA polymers: a reproducible porcine model of chronic total coronary occlusion. AB - We have hypothesized that oxygen enhanced three-dimensional microporous poly-L lactic acid (L-PLA) bioabsorbable polymer constructs could be implanted to produce a subacute occlusion in a porcine coronary artery, forming a thrombofibrotic occlusion containing microvascular channels. Chronic total occlusion (CTO) is increasingly prevalent in patients who present for percutaneous interventions. No reproducible animal coronary model simulating human CTOs has previously been developed. Swine coronary arteries were cannulated and a microporous L-PLA polymer pledget was advanced into a preselected segment of coronary. The coronary arteries were angiographically re-imaged at day 3, day 10, and day 28, to document the presence or absence of an occlusion. Histopathology was also performed at each time point to evaluate the lesion characteristics. A novel three-dimensional L-PLA microporous polymer construct, when implanted into porcine coronary arteries, reproducibly results in the development of a CTO at day 3. The histopathology in this porcine coronary model of CTO at day 28 closely mimics human coronary CTO, including the presence of microvascular channels and dense collagen and elastic tissue in the occlusion. PMID- 16475165 TI - Knockdown of NFAT3 blocked TPA-induced COX-2 and iNOS expression, and enhanced cell transformation in Cl41 cells. AB - The nuclear factor of activated-T-cells (NFAT) family is a ubiquitous transcription factor that mediates regulation on various gene expressions. Recent studies indicate that NFAT may implicate in cancer process, mainly through its direct regulation on the cyclooxygenase-2 (COX-2) gene expression. There is also evidence suggesting another aspect of NFAT in tumor suppression. However, the according mechanism remains unknown. In this study, we used a small interfering RNA (siRNA) expression construct to study the role of NFAT3 in the 12-O tetradecanoylphorbol-13-acetate (TPA)-induced cell transformation with the tumor promotion-sensitive mouse epidermal Cl41 cells. Our results showed that TPA was able to induce NFAT3 activation in Cl41 cells. Stable transfection of NFAT3 siRNA specifically reduced endogenous NFAT3 expression. At the same time, TPA-induced expression of both COX-2 and inducible nitric oxide synthase (iNOS) were blocked. However, anchorage-independent transformation in response to TPA was significantly enhanced in NFAT3 siRNA stable transfectants as compared with vector transfectants. Moreover, treatment with the iNOS specific inhibitor aminoguanidine (AG) also enhanced Cl41 cells transformation induced by TPA. As COX-2 expression is proved to be required for cell transformation in Cl41 cells in our recent studies, our results demonstrate that the inducible NFAT3-mediated iNOS upregulation represents a novel potent tumor-suppressing pathway and may contribute to the tumor suppressor functions of NFAT protein. PMID- 16475166 TI - The vertebrate E1/U17 small nucleolar ribonucleoprotein particle. AB - Each of the many different box H/ACA ribonucleoprotein particles (RNPs) present in eukaryotes and archaea consists of four common core proteins and one specific H/ACA small RNA, which bears the sequence elements H (ANANNA) and ACA. Most of the H/ACA RNPs are small nucleolar RNPs (snoRNPs), which are localized in nucleoli, and are one of the two major classes of snoRNPs. Most H/ACA RNPs direct pseudouridine synthesis in pre-rRNA and other RNAs. One H/ACA small nucleolar RNA (snoRNA), vertebrate E1/U17 (snR30 in yeast), is required for pre-rRNA cleavage processing that generates mature 18S rRNA. E1 snoRNA is encoded in introns of protein-coding genes, and the evidence suggests that human E1 RNA undergoes uridine insertional RNA editing. The vertebrate E1 RNA consensus secondary structure shows several features that are absent in other box H/ACA snoRNAs. The available UV-induced RNA-protein crosslinking results suggest that the E1 snoRNP is asymmetrical in vertebrate cells, in contrast to other H/ACA snoRNPs. The vertebrate E1 snoRNP in cells is surprisingly complex: (i) E1 RNA contacts directly and specifically several proteins which do not appear to be any of the H/ACA RNP four core proteins; and (ii) multiple E1 RNA sites are needed for E1 snoRNP formation, E1 RNA stability, and E1 RNA-protein direct interactions. PMID- 16475167 TI - Paneling human thyroid cancer cell lines for candidate proteins for targeted anti angiogenic therapy. AB - Tumor angiogenesis is believed to result from an imbalance of pro- and anti angiogenic factors, some of which are candidates for targeted therapy. Such therapy has raised hopes for patients with undifferentiated thyroid carcinomas, who are facing a grave prognosis with a survival of only months. In this study, in vivo growth of xenografted human thyroid carcinomas unexpectedly responded quite differently to neutralizing anti-vascular endothelial growth factor (VEGF) antibody. In particular, lasting inhibition as well as accelerated growth occurred after treatment. Consequently, a panel of anti-angiogenic factors was addressed in a representative sample of thyroid carcinoma lines. VEGF, fibroblast growth factor (FGF-2), and endostatin were demonstrated by Western blotting and EIA, whereas PDGF-A, PDGF-B, and IL-6 were negative. Quantification of VEGF, FGF 2, and endostatin revealed a wide range of concentrations from 500 to 4,200 pg/ml VEGF, 5 to 60 pg/ml FGF-2, and 50 to 300 pg/ml endostatin, not related to a particular histologic thyroid carcinoma background. Angiostatin (kringles 1-3) was detected in all, but one of the cell lines. Finally, aaATIII was confirmed in FTC133 cells. These data highlight the complex regulation of angiogenesis in thyroid carcinoma cell lines and suggest that the array of angiogenic factors differs markedly between individual cell lines. For the first time, angiostatin, endostatin, and possibly also aaATIII are identified as novel candidate regulators of angiogenesis in thyroid carcinoma cells. PMID- 16475171 TI - The use of multiple "buddies" during transradial angioplasty in a complex calcified coronary tree. AB - We report our experience in transradial coronary angioplasty on a tortuous, calcified coronary tree in a patient who had been turned down for surgical revascularization. In view of the complexity of the coronary anatomy, successful completion of the procedure required plaque modification with the use of rotational atherectomy in the left anterior descending coronary artery, and the use of an inflated buddy balloon in the circumflex artery. The latter represents a novel, simple, and inexpensive technique, which may allow successful advancing of coronary stents in calcified and tortuous coronary arteries. PMID- 16475168 TI - Secretion of tartrate-resistant acid phosphatase by osteoclasts correlates with resorptive behavior. AB - There have been dramatic advances recently in our understanding of the regulation of osteoclastic differentiation. However, much less is known of the mechanisms responsible for the induction and modulation of resorptive behavior. We have developed a strategy whereby osteoclasts can be generated in vitro and released into suspension in a fully-functional state. We now exploit this approach to show that tartrate-resistant acid phosphatase (TRAP) is released by osteoclasts during bone resorption. TRAP release was inhibited by the secretion-inhibitor Brefeldin A, and was not accompanied by LDH release. This suggests that TRAP release is due to secretion, rather than cell death. Consistent with this, TRAP secretion was stimulated by resorbogenic cytokines, was inhibited by the resorption-inhibitor calcitonin, and correlated with excavation of the bone surface. We found that, in contrast to incubation on bone, incubation on plastic, glass, or vitronectin coated plastic substrates did not induce secretion of TRAP. This suggests that the induction of resorptive behavior in osteoclasts depends upon stimulation by bone matrix of a putative osteoclastic "mineral receptor." Release of TRAP by osteoclasts thus represents not only a productive approach to the analysis of the mechanisms that modulate the rate of resorptive activity, but also a system whereby the mechanism through which bone substrates induce resorptive behavior can be identified. PMID- 16475172 TI - Inadvertent stenting of patent ductus arteriosus with Amplatzer Vascular Plug. AB - We report a 12-month old patient who presented for murmur evaluation after percutaneous closure of type C patent ductus arteriosus (PDA) using a 10 mm Amplatzer Vascular Plug (AGA Medical Corporation, Golden Valley, MN) at an outside institution. Echocardiography revealed a large left-to-right shunt through the implanted device, inadvertently stenting the PDA instead of closing it. The patient underwent repeat catheterization with successful coil implantation within the Amplatzer Vascular Plug, completely eliminating the large residual ductal shunt. Although challenging, this case illustrates the technique of implanting coils within this occlusion device. This case also illustrates that occlusion of type C PDA utilizing the Amplatzer Vascular Plug may not only result in incomplete occlusion but also create a potentially worse clinical situation in which the PDA is stretched larger and stented open. Without consideration of simultaneous coil implantation within this device, use of the Amplatzer Vascular Plug might be contraindicated in type C PDA, because there may be no way to ensure successful closure by the Vascular Plug alone. PMID- 16475174 TI - The use of multiple coronary guidewires to occupy secondary branch vessels and facilitate PCI of a complex aneurysmal stenosis. AB - Percutaneous revascularization of complex coronary stenosis is dependent on establishing suitable guidewire position in the vascular bed distal to the lesion. We report the use of multiple 0.0014'' coronary guidewires to occupy unintended branch vessels and to facilitate PCI of a high-grade left circumflex obtuse marginal branch lesion with post-stenotic aneurysmal dilation of the vessel and multiple branch vessels. This technique enabled successful placement of a nitinol hydrophilic-coated wire into the distal vascular bed beyond the region of obstruction followed by placement of a drug-eluting stent with restoration of luminal dimensions and TIMI-3 angiographic flow. PMID- 16475173 TI - Repression of HIP/RPL29 expression induces differentiation in colon cancer cells. AB - We had previously shown that the expression of heparin/heparan sulfate interacting protein/ribosomal protein L29 (HIP/RPL29) was upregulated in colon cancer tissues. The present study investigated the role of HIP/RPL29 in differentiation in colon cancer cells. Inducing cellular differentiation in HT-29 cells by both sodium butyrate and glucose deprivation resulted in a significant downregulation of HIP/RPL29 expression. The beta-catenin/Tcf-4 pathway is the most important pathway controlling the switch between cellular differentiation and proliferation in intestinal epithelial cells. Inducing differentiation by dominant-negative inhibition of the beta-catenin/Tcf-4 complexes in LS174T cells also resulted in downregulation of HIP/RPL29. To determine whether a lower expression of HIP/RPL29 could induce differentiation in cancer cells, small interfering RNA (siRNA) targeting HIP/RPL29 was transfected into LS174T cells. The resultant knockdown of HIP/RPL29 expression induced cellular differentiation, as shown by the increased expression of two known markers of differentiation in LS174T cells, galectin-4 and mucin-2. In addition, the differentiation process induced by repression of HIP/RPL29 expression was accompanied by the upregulation of p21 and p53. In conclusion, HIP/RPL29 plays a role in the cellular differentiation process in colon cancer cells. The differentiation process is at least partially mediated by the upregulation of p21 and p53 pathways. PMID- 16475175 TI - Bronchodilator effect of salbutamol from two different spacer devices. AB - Our aim was to compare the bronchodilator effect of salbutamol delivered via a new holding chamber (Volumatic Soft, VS) with that of an established device (Volumatic, V) in asthmatic children. Children with stable asthma were recruited. They inhaled 100 microg, and 10 min later, 300 microg of salbutamol aerosol delivered via VS or V on day 1, and vice versa on day 2. Spirometry was measured at baseline, 10 min after 100 microg, and 15 min after 300 microg of salbutamol were given. The preference for either device was assessed by visual analogue score. Forty-four children with a median age of 9.2 years (interquartile range, 8.0-10.7) completed the study. There were significant improvements in forced expired volume in 1 sec (FEV1) with time throughout the study period for both V and VS (linear and quadratic trend P < 0.001). There was a statistically significant difference in postbronchodilator FEV1 between V and VS (P = 0.013). VS gave an overall greater change in FEV1 than V, by 1.8%. The preference scores for V and VS were 7.0 (IQR 5.0-8.0) and 9.0 (IQR 8.0-10.0) (p < 0.0005), respectively. In conclusion, comparable clinical efficacy was found for V and VS with respect to changes in FEV1 after salbutamol. Patients also showed a strong preference for the new device. PMID- 16475176 TI - Pulmonary alveolar proteinosis associated with anti-GM-CSF antibodies in a child: successful treatment with inhaled GM-CSF. PMID- 16475177 TI - Multislice computed tomography as an adjunct to the management of an in-stent stenosis in an infant with congenital heart disease: imaging for the future. AB - The case report demonstrates the value of multslice computed tomography for stent imaging and detection of an in-stent stenosis in an infant with pulmonary atresia and ventricular septal defect after stent placement in the right pulmonary artery after biventricular repair. PMID- 16475178 TI - Coherent corticomuscular oscillations originate from primary motor cortex: evidence from patients with early brain lesions. AB - Coherent oscillations of neurons in the primary motor cortex (M1) have been shown to be involved in the corticospinal control of muscle activity. This interaction between M1 and muscle can be measured by the analysis of corticomuscular coherence in the beta-frequency range (beta-CMCoh; 14-30 Hz). Largely based on magnetoencephalographic (MEG) source-modeling data, it is widely assumed that beta-CMCoh reflects direct coupling between M1 and muscle. Deafferentation is capable of modulating beta-CMCoh, however, and therefore the influence of reafferent somatosensory signaling and corresponding neuronal activity in the somatosensory cortex (S1) has been unclear. We present transcranial magnetic stimulation (TMS) and MEG data from three adult patients suffering from congenital hemiparesis due to pre- and perinatally acquired lesions of the pyramidal tract. In these patients, interhemispheric reorganization had resulted in relocation of M1 to the contralesional hemisphere, ipsilateral to the paretic hand, whereas S1 had remained in the lesioned hemisphere. This topographic dichotomy allowed for an unequivocal topographic differentiation of M1 and S1 with MEG (which is not possible if M1 and S1 are directly adjacent within one hemisphere). In all patients, beta-CMCoh originated from the contralesional M1, in accordance with the TMS-evoked motor responses, and in contrast to the somatosensory evoked fields (SEFs) for which the sources (N20m) were localized in S1 of the lesioned hemisphere. These data provide direct evidence for the concept that beta-CMCoh reflects the motorcortical efferent drive from M1 to the spinal motoneuron pool and muscle. No evidence was found for a relevant contribution of neuronal activity in S1 to beta-CMCoh. PMID- 16475179 TI - Radical reaction C3H+NO: a mechanistic study. AB - Although a number of hydrocarbon radicals including the heavier C(3)-radicals C(3)H(3) and C(3)H(5) have been experimentally shown to deplete NO effectively, no theoretical or experimental attempts have been made on the reactivity of the simplest C(3)-radical towards NO. In this article, we report our detailed mechanistic study on the C(3)H+NO reaction at the Gussian-3//B3LYP/6-31G(d) level by constructing the singlet and triplet electronic state [H,C(3),N,O] potential energy surfaces (PESs). The l-C(3)H+NO reaction is shown to barrierlessly form the entrance isomer HCCCNO followed by the direct O-elimination leading to HCCCN+(3)O on triplet PES, or by successive O-transfer, N-insertion, and CN bond rupture to generate the product (1)HCCN+CO on singlet PES. The possible singlet triplet intersystem crossings are also discussed. Thus, the novel reaction l C(3)H+NO can proceed effectively even at low temperatures and is expected to play an important role in both combustion and interstellar processes. For the c C(3)H+NO reaction, the initially formed H-cCCC-NO can most favorably isomerize to HCCCNO, and further evolution follows that of the l-C(3)H+NO reaction. Quantitatively, the c-C(3)H+NO reaction can take place barrierlessly on singlet PES, yet it faces a small barrier 2.7 kcal/mol on triplet PES. The results will enrich our understanding of the chemistry of the simplest C(3)-radical in both combustion and interstellar processes, which to date have received little attention despite their importance and available abundant studies on its structural and spectroscopic properties. PMID- 16475180 TI - Intracoronary stent dislodgment: updated strategy enabled by the new generation of materials. AB - Stent dislodgment remains a vexing problem in percutaneous coronary intervention. The technological improvements in machine-crimped premounted stents have led to a reduced incidence of dislodgment but may have exacerbated the problem of stent dislodgment (SD). Previous reports describing the management of SD have concentrated predominantly on manually mounted stents. We present a case where the position of a dislodged premounted stent in the distal left main stem was unfavorable for previously described solutions. The problem was elegantly resolved by taking advantage of the improved performance of the current generation of wires and balloons to achieve a rapid first-rate clinical outcome. PMID- 16475181 TI - Theoretical mechanistic study on the radical-molecule reaction of CHCl2/CCl3 with NO2. AB - The radical-molecule reaction mechanism of CHCl(2) and CCl(3) with NO(2) have been explored theoretically at the B3LYP/6-311G(d,p) and MC-QCISD (single-point) levels. For the singlet potential energy surface (PES) of CHCl(2) + NO(2) reaction, the association of CHCl(2) with NO(2) was found to be a barrierless carbon-to-nitrogen approach forming an energy-rich adduct a (HCl(2)CNO(2)) followed by isomerization to b(1) (trans-cis-HCl(2)CONO), which can easily interconvert to b(2), b(3), and b(4). Subsequently, the most feasible pathway is the 1,3-chlorine migration associated with N-O1 bond cleavage of b(1) leading to P(1) (CHClO + ClNO). The second competitive pathway is the 1,4-chlorine migration along with N-O1 bond rupture of b(4) giving rise to P(2) (CHClO + ClON). Moreover, some of P(1) and P(2) can further dissociate to give P(6) (CHClO + Cl + NO). The lesser followed competitive channel is the 1,3-H-shift from C to N atom along with N-O1 bond rupture of b(1) to form P(3) (CCl(2)O + HNO). The concerted 1,4-H-shift accompanied by N-O1 bond fission of b(3) to product P(4) (CCl(2)O + HON) is even much less feasible. For the singlet PES of CCl(3) + NO(2) reaction, the only primary product is found to be P(1) (CCl(2)O + ClNO), which can lead to P(2) (CCl(2)O + Cl + NO) via dissociation of ClNO. The obtained major products CHClO and CCl(2)O for CHCl(2) + NO(2) and CCl(3) + NO(2) reactions, respectively, are in good agreement with kinetic detection in experiment. Compared with the singlet pathways, the triplet pathways may have less contributions to both reactions. Because the rate-determining transition state involved in the feasible pathways lie above the reactants R, the title reactions may be important in high temperature processes. The similarities and discrepancies among the CH(n)Cl(3-n) + NO(2) (n == 0-2) reactions are discussed in terms of the substitution effect. The present study may be helpful for further experimental investigation of the title reactions. PMID- 16475182 TI - Crystal structure of a putative HTH-type transcriptional regulator yxaF from Bacillus subtilis. PMID- 16475183 TI - Nonrecurring transient cortical blindness after coronary angiography: a role for hypoventilation and hypercarbia? AB - A 73-year-old morbidly obese patient suffered cortical blindness following coronary angiography. Symptoms rapidly resolved and did not recur following a repeated procedure. Hypoventilation due to narcotics with subsequent hypercarbia and blood-brain barrier breaching may be implicated in the pathogenesis of this complication. PMID- 16475184 TI - SUMO regulates the cytoplasmonuclear transport of its target protein Daxx. AB - It is known that Fas death domain-associated protein (Daxx) possesses both putative nuclear and cytoplasmic functions. However, the nuclear transport mechanism is largely unknown. This study examined the nuclear location signal (NLS) of Daxx and whether the nuclear transport of Daxx was mediated by small ubiquitin-related modifier (SUMO). Two NLS motifs of Daxx, leucine (L)-rich nuclear export signal (NES)-like motif (188IXXLXXLLXL197) and C-terminal lysine (K) rich NLS2 (amino acids 627-634) motif, were identified and the K630 and K631 on the NLS2 motif were characterized as the major sumoylation sites of Daxx by in vitro sumoylation analysis. Proteins of inactive SUMO (SUMO-delta), a sumoylation incompetent mutant, and Daxx NLS mutants (Daxx-NES(mut) and Daxx NLS2(mut)) were dispersed in cytoplasm. The cytoplasmic dispersed Daxx mutants could be relocalized to nucleus by cotransfection with active SUMO, but not with inactive SUMO-delta, demonstrating the role of SUMO on regulating the cytoplasmonuclear transport of Daxx. However, inactive SUMO-delta could also be relocalized to nucleus during cotransfection with wild-type Daxx, suggesting that SUMO regulation of the cytoplasmonuclear transport of its target protein Daxx does not need covalent modification. This study shows that cytoplasmic SUMO has a biological role in enhancing the cytoplasmonuclear transport of its target protein Daxx and it may be done through the non-sumoylation interactions. PMID- 16475185 TI - Gap junction communications influence upon fibroblast synthesis of Type I collagen and fibronectin. AB - In rats polyvinyl alcohol sponge subcutaneous implants treated with gap junctional intercellular communications (GJIC) uncouplers showed reduced deposition of connective tissue. Do uncouplers inhibit the synthesis and deposition of a new connective tissue by fibroblasts? Confluent human dermal fibroblasts in serum-free medium received either endosulfan or oleamide, GJIC uncouplers. Collected media were subjected to Dot Blot analysis for native Type I collagen and fibronectin. Uncoupler-treated fibroblasts released less Type I collagen, while there was no change in fibronectin release. Collagen synthesis was restored to normal, when the uncouplers were removed, showing that these uncouplers were reversible and not toxic to cells. Northern blot analysis revealed procollagen alpha1 (I) mRNA was minimally affected by endosulfan. Oleamide-treated 17-day chick embryo calvaria explants were incubated with Type I collagen antibody, frozen, cryosectioned, and then subjected to rhodamine (Rh) tagged anti-mouse-IgG antibody, to detect newly deposited Type I collagen. Fluorescent antibody-collagen complexes were localized on the periphery of cells in control calvaria, but absent around cells in oleamide-treated calvaria. GJIC optimize collagen synthesis but not fibronectin synthesis. The lack of connective tissue deposited in granulation tissues treated with uncouplers appears related to the inhibition of collagen synthesis. These findings suggest that altering GJIC might control collagen deposition in scarring. PMID- 16475186 TI - Percutaneous coronary intervention for extrinsic coronary compression after pulmonary valve replacement. AB - Coronary artery compression is a rare and potentially fatal complication after pulmonary valve replacement. This report describes myocardial infarction from extrinsic left main coronary artery compression after pulmonary valve replacement in a 10-y-old boy. He was successfully treated with percutaneous coronary intervention. PMID- 16475187 TI - Fibromuscular dysplasia of renal arteries: percutaneous revascularization based on hemodynamic assessment with a pressure measurement guidewire. AB - Fibromuscular dysplasia (FMD) leading to renal artery stenosis and hypertension is one of the most common treatable causes of secondary hypertension. However, frequently it can be difficult to judge the anatomical severity of a stenotic lesion with various noninvasive and invasive imaging modalities. We present two patients with poorly controlled hypertension and FMD affecting the renal arteries, in whom there were no anatomically significant stenoses by renal magnetic resonance angiography or selective renal artery angiography. Utilizing a 0.014'' high fidelity micromanometer tipped PressureWire XT (Radi, Reading, MA), to measure intravascular pressure gradients throughout the diseased renal arteries, we identified physiologically significant stenoses, and successfully treated both patients with percutaneous transluminal angioplasty. PMID- 16475188 TI - Festschrift in honor of the 65th birthday of George D. Rose. PMID- 16475189 TI - Anchoring technique to improve guiding catheter support in coronary angioplasty of chronic total occlusions. AB - The primary reason for unsuccessful angioplasty of chronic total occlusions (CTOs) is an inability to pass the guidewire through the occlusion. Optimal guiding catheter support is a prerequisite for successful angioplasty of CTO. We performed guidewire manipulation by anchoring a balloon in a side-branch vessel in order to achieve adequate guiding catheter support. With this novel anchoring technique, we successfully achieved guidewire passage through the CTO. PMID- 16475190 TI - Restenosis of a sirolimus drug-eluting stent: aorto-ostial involvement of the proximal right coronary artery. AB - We present a case of restenosis of a sirolimus drug-eluting stent in the ostium of the right coronary artery due to severe crimping and underexpansion of the stent in an area of heavy calcification, detected by intracoronary ultrasound but not by angiography. We discuss issues relevant to management of the restenotic drug-eluting stent in the aorto-ostial location. PMID- 16475191 TI - Functional grouping based on signatures in protein termini. AB - The two ends of each protein are known as the amino (N-) and carboxyl (C-) termini. Short signatures in a protein's termini often carry vital cellular function. No systematic research has been conducted to address the importance of short signatures (3 to 10 amino acids) in protein termini at the proteomic level. Specifically, it is unknown whether such signatures are evolutionarily conserved, and if so, whether this conservation confers shared biological functions. Current signature detection methods fail to detect such short signatures due to inadequate statistical scores. The findings presented in this study strongly support the notion that functional significance of protein sets may be captured by short signatures at their termini. A positional search method was applied to over one million proteins from the UniProt database. The result is a collection of about a thousand significant signature groups (SIGs) that include previously identified as well as many novel signatures in protein termini. These SIGs represent protein sets with minimal or no overall sequence similarity excepting the similarity at their termini. The most significant SIGs are assigned by their strong correspondence to functional annotations derived from external databases such as Gene Ontology. Each of the SIGs is associated with the statistical significance of its functional association. These SIGs provide a valuable source for testing previously overlooked signatures in protein termini and allow for the investigation of the role played by such signatures throughout evolution. The SIGs archive and advanced search options are available at http://www.proteus.cs.huji.ac.il. PMID- 16475192 TI - Morphometric discrimination of early life stage Lampetra tridentata and L. richardsoni (Petromyzonidae) from the Columbia River Basin. AB - The effectiveness of morphometric and meristic characteristics for taxonomic discrimination of Lampetra tridentata and L. richardsoni (Petromyzonidae) during embryological, prolarval, and early larval stages (i.e., age class 1) were examined. Mean chorion diameter increased with time from fertilization to hatch and was significantly greater for L. tridentata than for L. richardsoni at 1, 8, and 15 days postfertilization. Lampetra tridentata larvae had significantly more trunk myomeres than L. richardsoni; however, trunk myomere numbers were highly variable within species and deviated from previously published data. Multivariate examinations of prolarval and larval L. tridentata (7.2-11.0 mm; standard length) and L. richardsoni (6.6-10.8 mm) were conducted based on standard length and truss element lengths established from eight homologous landmarks. Principal components analysis indicated allometric relationships among the morphometric characteristics examined. Changes in body shape were indicated by groupings of morphometric characteristics associated with body regions (e.g., oral hood, branchial region, trunk region, and tail region). Discriminant function analysis using morphometric characteristics was successful in classifying a large proportion (> 94.7%) of the lampreys sampled. PMID- 16475193 TI - Commentary on: "Assembly of a tetrameric alpha-helical bundle: computer simulations on an intermediate-resolution protein model" [Proteins 2001;44:376 391]. PMID- 16475196 TI - Reconciling the "old" and "new" views of protein allostery: a molecular simulation study of chemotaxis Y protein (CheY). AB - A combination of thirty-two 10-ns-scale molecular dynamics simulations were used to explore the coupling between conformational transition and phosphorylation in the bacteria chemotaxis Y protein (CheY), as a simple but representative example of protein allostery. Results from these simulations support an activation mechanism in which the beta4-alpha4 loop, at least partially, gates the isomerization of Tyr106. The roles of phosphorylation and the conserved Thr87 are deemed indirect in that they stabilize the active configuration of the beta4 alpha4 loop. The indirect role of the activation event (phosphorylation) and/or conserved residues in stabilizing, rather than causing, specific conformational transition is likely a feature in many signaling systems. The current analysis of CheY also helps to make clear that neither the "old" (induced fit) nor the "new" (population shift) views for protein allostery are complete, because they emphasize the kinetic (mechanistic) and thermodynamic aspects of allosteric transitions, respectively. In this regard, an issue that warrants further analysis concerns the interplay of concerted collective motion and sequential local structural changes in modulating cooperativity between distant sites in biomolecules. PMID- 16475197 TI - Proteomics: clinical applications for head and neck squamous cell carcinoma. AB - The emerging field of proteomics offers great promise for unraveling the complex molecular events of tumorigenesis, as well as those that control clinically important tumor behaviors such as metastases, invasion, and resistance to therapy. Understanding the molecular basis of these tumor characteristics will usher in a new age of individualized cancer therapy. Advances in genomics, the study of genes and gene expression, as well as technologic advances have made possible study of all expressed proteins or the proteome (proteomics). Whereas genomics offers the opportunity to examine gene expression or variation in gene sequence, proteomics encompasses evaluation of protein expression, activation, modification, and degradation and ambitiously targets protein function. Although the present age has already been referred to as the post-genomic era, it is our opinion that both proteomics and genomics will provide clinically useful and complementary information that will speed scientific understanding of cancer. Ultimately, these new technologies will positively impact care of patients with head and neck squamous cell carcinoma. PMID- 16475198 TI - Squamous cell carcinoma of the parotid gland. AB - BACKGROUND: Our objective was to evaluate the outcome of patients treated for squamous cell carcinoma (SCC) of the parotid gland. METHODS: We conducted a retrospective chart review of the tumor registry from 1982 through 2003 at a tertiary referral medical center. Patients with SCC of the parotid gland were identified and followed for a minimum of 2 years after therapy. RESULTS: SCC involving the parotid was identified in 66 patients. The tumor was a metastasis from a known primary site in 41 patients (62%). In 16 patients (24%), no other primary site was identified, and the tumor may have originated in the parotid gland. Nine patients (14%) were undetermined. Therapy frequently included surgery. The integrity of the facial nerve was preserved in 92% of surgical patients. Only eight patients initially had clinical evidence of cervical metastasis; however, cervical metastasis was identified in 25 patients (44%), changing the course of therapy. CONCLUSION: SCC of the parotid gland was metastatic from a known primary tumor in more than half of the patients. The most common site of the primary tumor was a cutaneous malignancy of the head and neck. The high incidence of cervical lymph node involvement underscores the diagnostic and therapeutic importance of neck dissection with parotidectomy. PMID- 16475199 TI - Advanced-staged tonsillar squamous carcinoma: organ preservation versus surgical management of the primary site. AB - BACKGROUND: Our aim was to review our experience in the management of advanced tonsillar squamous cell carcinoma (SCC) and to compare treatment outcomes between patients treated with and without surgery to the primary site. METHODS: The records of 74 patients with advanced-stage tonsillar SCC were reviewed. The median age at diagnosis was 58 years. Thirty-eight patients received definitive surgery to the primary site, and 36 were treated with an organ-preservation approach (OP) using radiotherapy +/- chemotherapy. RESULTS: No significant difference in overall survival (OS) or freedom from relapse (FFR) by treatment was found. T classification and N status were significant independent predictors on multivariate analysis for OS and FFR. Major late toxicity was noted in 10 patients in the surgical group and nine in the OP group. CONCLUSION: Patients treated with OP and primary surgery had comparable OS and FFR. T classification and N status were significant independent predictors for tumor relapse and survival. On the basis of these results, we favor organ-preservation therapy for patients with advanced-stage tonsillar SCC. PMID- 16475200 TI - New modification of the mandibulotomy approach without lip splitting. AB - BACKGROUND: A lower lip-splitting incision has traditionally been performed with different types of mandibulotomy approaches for obtaining wide access to oral and oropharyngeal cancers. However, lip splitting can be associated with unfavorable aesthetic results. We describe our new modification of a traditional mandibulotomy approach without lip splitting to avoid these morbidities. This is a case series in a tertiary referral center. METHODS: The primary tumor site was the oropharynx in four cases, the oral cavity in two cases, and the parapharynx in one case. Each case was assessed for TNM staging, perioperative complications, status of the resection margins, tumor recurrence, and the aesthetic and functional results of the lower lip. RESULTS: All the tumors were safely removed by means of our modified non-lip-splitting mandibulotomy approach through the combined intraoral and transcervical routes with adequate resection margins. There were no troublesome difficulties in reconstruction of the surgical defects with various major flaps. The cosmetic results were excellent with intact lip function. CONCLUSION: We believe this new modified non-lip-splitting mandibulotomy approach could replace the conventional mandibulotomy approach for some selected malignant lesions, with excellent cosmetic and functional results of the lower lip. PMID- 16475201 TI - Cardiovascular and respiratory complications after major head and neck surgery. AB - BACKGROUND: Our aim was to gain insight into the incidence rates for, distribution of, and risk factors of postoperative cardiovascular and respiratory complications in major head and neck surgery. METHODS: We performed a retrospective review of 469 patients who had undergone primary major head and neck surgery. Outcome measures were incidence rates, risk factors, and distribution over time for postoperative cardiovascular and respiratory complications. A multivariate analysis was performed. RESULTS: The incidence rates for cardiovascular and respiratory complications were 57 of 469 (12%) and 50 of 469 (11%), respectively. The incidence rate for heart failure exceeded that for pneumonia. The peak incidence for cardiovascular complications was on the first postoperative day; for respiratory complications, on the second postoperative day. Risk factors for cardiovascular complications were age, pulmonary disease, alcohol abuse, and tumor location; risk factors for respiratory complications were pulmonary disease, previous myocardial infarction, and American Society of Anesthesiologists (ASA) grade. CONCLUSION: In this study, the incidence rates for cardiovascular and respiratory complications were very similar. The first postoperative day was crucial with regard to cardiovascular complications. Age and chronic pulmonary diseases were the common risk factors for cardiovascular and respiratory complications. PMID- 16475202 TI - Influence of passive humidification on respiratory heat loss in tracheotomized patients. AB - BACKGROUND: The aim of this study was to evaluate changes in total respiratory heat loss during use of a heat and moisture exchanger (HME) in tracheotomized patients. METHODS: Tracheal humidity and temperature were measured before the application and during use of the HME (plastic foam impregnated with CaCl2), and total respiratory heat loss was calculated. RESULTS: No significant difference was found between the convective heat exchange before and after use of the HME for a 10-minute period. When the HME was placed on the tracheal opening, the evaporative heat exchange and the total respiratory heat loss decreased significantly. CONCLUSIONS: The results indicate that passive airway humidification is effective in tracheotomized patients even after a 10-minute period. However, the positive effect on the energy balance of the tracheal mucosa after prolonged use of the HME remains to be proven. PMID- 16475203 TI - Preservation of quality of life after intensity-modulated radiotherapy for early stage nasopharyngeal carcinoma: results of a prospective longitudinal study. AB - BACKGROUND: Xerostomia is a ubiquitous complication after conventional radiotherapy for nasopharyngeal carcinoma (NPC) that seriously impairs patient quality of life (QOL). The effect on QOL of parotid-sparing intensity-modulated radiotherapy (IMRT) for early-stage NPC was assessed prospectively. METHODS: Thirty-two patients with T1-2,N0-1,M0 NPC received IMRT. Saliva flow was measured, and the Medical Outcomes Short Form 36 (SF-36), European Organization for Research and Treatment of Cancer (EORTC) core, and EORTC head and neck module (QLQ-H&N35) questionnaires were completed at baseline and 2, 6, and 12 months after IMRT. RESULTS: Saliva flow recovered to at least 25% of baseline in most cases after 1 year. Physical function, fatigue, and vitality were problems at 2 months and then resolved (p < or = .002). Global health scales showed continuous improvement in QOL after treatment (p < or = .004). Xerostomia and sticky saliva were problems 2 months after treatment; thereafter, continuous improvement occurred (p < or = .001). Xerostomia scores correlated with general aspects of QOL (p < or = .044). CONCLUSION: IMRT for early-stage disease preserved key aspects of QOL, and a reduction in xerostomia symptoms enhanced broader aspects of QOL. PMID- 16475204 TI - Phase I/II study of S-1 combined with carboplatin in recurrent and/or metastatic head and neck cancer as outpatient chemotherapy. AB - BACKGROUND: The aims of this study were to determine the recommended dose (RD) in combination chemotherapy of S-1 and carboplatin in recurrent and/or metastatic head and neck cancer (phase I) and to examine response rate and toxicities at RD (phase II). METHODS: S-1 was given orally at a fixed dose of 40, 50, or 60 mg twice daily based on the patient's body surface area for 21 days followed by a 14 day rest. In phase I, carboplatin was given intravenously to a cohort of three patients on day 8 at a dose of 2.5, 3.0, 3.5, or 4.0 area under the curve (AUC) values, depending on the dose-limiting toxicities (DLTs). RESULTS: In phase I, the RD was estimated as 2.5 AUC. In phase II, thrombocytopenia and leukopenia were most commonly found as hematologic DLTs, which were manageable without hospitalization. The response rate was 40.9%. CONCLUSIONS: This regimen is considered to be active against recurrent and/or metastatic head and neck cancer in an outpatient setting. PMID- 16475205 TI - Expression of cancer testis antigens in head and neck squamous cell carcinomas. AB - BACKGROUND: There is considerable interest in the expression of cancer testis (CT) antigens in human cancers, because they may serve as the basis for diagnostic tests or an immunologic approach to therapy, or as prognostic markers. METHODS: On this basis, we evaluated by semiquantitative reverse-transcriptase polymerase chain reaction (RT-PCR) the expression of genes that code for tumor antigens (melanoma antigen-1 [MAGE-1], MAGE-4, MAGE-10, MAGE-12, B melanoma antigen, CTL-recognized antigen melanoma antigen (CT antigen 2) [LAGE], New York esophageal squamous cell carcinoma antigen (CT antigen 1) [NYESO-1], and preferentially expressed antigen of melanoma [PRAME]) in surgical samples of the tumors, margins, and lymph nodes (when present) from patients with a diagnosis of head and neck carcinoma. The study was conducted on 33 patients (31 men and two women), aged 31 to 94 years (mean, 56 years), with squamous cell carcinomas located in the mouth (15 cases), larynx (14 cases), and pharynx (four cases). RESULTS: The findings were compared with the clinical course and laboratory data. Expression of at least one antigen was observed in 66.6% of cases, with different rates of expression according to tumor staging (100% of T4, 57% of T3, 50% of T1 and T2) and smoking habit. There was a significantly higher expression of multiple genes (two or more) in tumors in advanced stages. CONCLUSIONS: We conclude that the tumor-specific antigen genes are expressed in variable frequencies and intensities in the primary lesions of head and neck squamous cell carcinomas and in their metastases, with expression of the PRAME gene being always present in the metastastatic lymph nodes. In primary lesions, gene expression correlated with smoking habit and with advanced tumors with a higher malignant potential, with the frequent expression of two or more of these genes. PMID- 16475206 TI - Structural MRI of carotid artery atherosclerotic lesion burden and characterization of hemispheric cerebral blood flow before and after carotid endarterectomy. AB - Collateral circulation plays a major role in maintaining cerebral blood flow (CBF) in patients with internal carotid artery (ICA) stenosis. CBF can remain normal despite severe ICA stenosis, making the benefit of carotid endarterectomy (CEA) or stenting difficult to assess. Before and after surgery, we assessed CBF supplied through the ipsilateral (stenotic) or contralateral ICA individually with a novel hemisphere-selective arterial spin-labeling (ASL) perfusion MR technique. We further explored the relationship between CBF and ICA obstruction ratio (OR) acquired with a multislice black-blood imaging sequence. For patients with unilateral ICA stenosis (n = 19), conventional bilateral labeling did not reveal interhemispheric differences. With unilateral labeling, CBF in the middle cerebral artery (MCA) territory on the surgical side from the ipsilateral supply (53.7 +/- 3.3 ml/100 g/min) was lower than CBF in the contralateral MCA territory from the contralateral supply (58.5 +/- 2.7 ml/100 g/min), although not statistically significant (p = 0.09). The ipsilateral MCA territory received significant (p = 0.02) contralateral supply (7.0 +/- 2.7 ml/100 g/min), while ipsilateral supply to the contralateral side was not reciprocated. After surgery (n = 11), ipsilateral supply to the MCA territory increased from 57.3 +/- 5.7 to 67.3 +/- 5.4 ml/100 g/min (p = 0.03), and contralateral supply to the ipsilateral MCA territory decreased. The best predictor of increased CBF on the side of surgery was normalized presurgical ipsilateral supply (r(2) = 0.62, p = 0.004). OR was less predictive of change, although the change in normalized contralateral supply was negatively correlated with OR(excess) (=OR(ipsilateral) - OR(contralateral)) (r(2) = 0.58, p = 0.006). The results demonstrate the effect of carotid artery stenosis on blood supply to the cerebral hemispheres, as well as the relative role of collateral pathways before surgery and redistribution of blood flow through these pathways after surgery. Unilateral ASL may better predict hemodynamic surgical outcome (measured by improved perfusion) than ICA OR. PMID- 16475207 TI - High risk of contralateral breast carcinoma in women with hereditary/familial non BRCA1/BRCA2 breast carcinoma. AB - BACKGROUND: The objectives of the current study were to estimate the risk of developing contralateral breast carcinoma (CBC) among women with hereditary/familial non-BRCA1/BRCA2 breast carcinoma and to determine the factors that may predict their risk of CBC. METHODS: The study sample consisted of all families (n = 217 families) that were referred between 1994-2001 to the Clinic of Cancer Genetics at the University Hospital of Umea for suspected hereditary breast carcinoma. The study included all women in the 217 families who had carcinoma of the breast as their first primary invasive malignancy diagnosed between 1970-2001 in northern Sweden. Exclusion criteria were an estimated lifetime risk < 20%, BRCA1/BRCA2 mutation, noninvasive carcinoma (ductal or lobular carcinoma in situ), and bilateral breast carcinoma. In the final analysis, 204 women were included from 120 families. RESULTS: The cumulative probability of developing CBC among women who had hereditary/familial non BRCA1/BRCA2 breast carcinoma after 20 years was 27.3% (95% confidence interval, 15.0-37.8) compared with the expected risk (4.9%) among women in northern Sweden who had primary breast carcinoma. A significantly increased risk of CBC was associated with age younger than 50 years at the time of diagnosis of the first primary breast carcinoma (P = 0.006). Adjuvant hormone therapy reduced the risk of CBC (P = 0.036). CONCLUSIONS: Women with hereditary/familial non-BRCA1/BRCA2 breast carcinoma had a high risk of developing CBC. This risk was attenuated further among women who were younger at the time of onset, who had a cumulative probability of developing CBC of nearly 40% after 15 years, which is similar to the estimated risk among BRCA1/BRCA2 mutation carriers. The results of this study emphasized the importance of genetic counseling for these women. PMID- 16475208 TI - Racial disparity and socioeconomic status in association with survival in older men with local/regional stage prostate carcinoma: findings from a large community based cohort. AB - BACKGROUND: Few studies have examined the outcomes for Hispanic men with prostate carcinoma and incorporated socioeconomic factors in association with race/ethnicity in affecting survival, adjusting for factors on cancer stage, grade, comorbidity, and treatment. METHODS: We studied a population-based cohort of 61,228 men diagnosed with local or regional stage prostate carcinoma at age 65 years or older between 1992 and 1999 in the 11 SEER (Surveillance, Epidemiology, and End Results) areas, identified from the SEER-Medicare linked data with up to 11 years of followup. RESULTS: Low socioeconomic status was significantly associated with decreasing survival in all men with prostate carcinoma. Those living in the community with the lowest quartile of socioeconomic status were 31% more likely to die than those living in the highest quartile (hazard ratio [HR] of all-cause mortality: 1.31; 95% confidence interval [CI]: 1.25-1.36) after adjustment for patient age, comorbidity, Gleason score, and treatment. The HR remained almost unchanged after controlling for race/ethnicity (HR: 1.32; 95% CI: 1.26-1.38). Compared with Caucasians, the risk of mortality in African American men was marginally significantly higher (HR: 1.06; 95% CI: 1.01-1.11) after controlling for education, and no longer significant after adjusting for poverty, income, or composite socioeconomic variable; the HR was lower for Hispanic men (HR: 0.80; 95% CI: 0.72-0.89) after adjustment for education and other socioeconomic variables. CONCLUSION: Racial disparity in survival among men with local or regional prostate carcinoma was largely explained by socioeconomic status and other factors. Lower socioeconomic status appeared to be one of the major barriers to achieving comparable outcomes for men with prostate carcinoma. PMID- 16475209 TI - Neuroblastoma in adolescents: the Italian experience. AB - BACKGROUND: Neuroblastoma (NB) occurs rarely during adolescence, and information is scarce on its characteristics and clinical course in this age group. METHODS: Patients with NB who were included in the Italian Neuroblastoma Registry were considered for the current study. The clinical characteristics and survival of adolescents (age at diagnosis between 10 yrs and 18 yrs) were compared with those of children (ages 1-9 yrs). Infants (age < 1 yr) were excluded because of their well known favorable clinical course. RESULTS: Between 1116 children and 53 adolescents who were evaluated, no differences were documented with regard to the primary tumor site and the prevalence of advanced stage at diagnosis. If only patients with Stage IV NB were considered, then adolescents were less likely to be diagnosed with bone/bone marrow metastases (77%) compared with children (94%; P = 0.038), but adolescents were more likely to have metastases at unusual sites, such as the lung parenchyma or the central nervous system (23% vs. 7%, respectively; P = 0.005). With regard to biologic characteristics, adolescents did not differ significantly from children, although they always had a lower prevalence of unfavorable markers. In particular, MYCN amplification was documented in 21% of children and in 11% of adolescents (P = 0.173). At age 10 years, adolescents had a 20% overall survival rate and a 22% event-free survival rate. Adolescents who had resectable disease had a 73% overall survival rate, which was worse compared with the rate among children with the same disease stage (89%), although the difference did not reach statistical significance (P = 0.159). No differences in survival were observed among patients with Stage IV NB, and adolescents had a probability of survival almost identical to that among children (6% vs. 16%, respectively; P = 0.481). However, when the analysis was restricted to events that occurred after patients developed a recurrence, even if the final outcome was poor for both groups, the difference was statistically significant (P = 0.022) mostly because of the more indolent disease course observed among the adolescents. This effect was even more evident for patients with Stage IV NB. When the 6-year cut-off point was used to separate children from adolescents, a significantly worse overall survival rate (P = 0.036) was documented for adolescents who had resectable disease (81% vs. 93% in children). CONCLUSIONS: NB in adolescents had clinical and biologic characteristics similar to those observed among children. The clinical course of NB probably is correlated significantly with age at diagnosis, but information is scarce on the role of the biologic risk factors in this age group. The authors were able to identify a group of patients with a cut-off age between 6 years and 10 years that had a more indolent course but a worse prognosis. PMID- 16475210 TI - Accumulation of DNA methylation is associated with tumor stage in gastric cancer. AB - BACKGROUND: The authors purpose in this study was to clarify the difference in terms of clinicopathologic features between gastric cancer (GC) with high numbers of DNA methylated genes and CpG island methylator phenotype (CIMP)-positive GC as originally defined. METHODS: We analyzed DNA methylation of 12 tumor-related genes (hMLH1, MGMT, p16(INK4a), CDH1, RAR-beta, HLTF, RIZ1, TM, FLNc, LOX, HRASLS, HAND1) in 75 samples of GC from 75 patients, 25 samples of corresponding nonneoplastic mucosa from 25 patients, and 10 samples of normal gastric mucosa from 10 healthy young individuals by methylation-specific polymerase chain reaction (PCR) and bisulfite PCR. We also investigated CIMP status by examining the methylation of five MINT loci and p53 mutation status by PCR single-strand conformation polymorphism. We measured levels of expression of mRNAs for these 12 genes by quantitative reverse transcription PCR in 50 GC specimens. RESULTS: The average number of methylated genes per tumor was 4.83. DNA methylation of each gene was correlated with low expression of the respective mRNA. High methylation (GC with 5 or more methylated genes) was detected in 39 (52.0%) of 75 GCs. Twenty nine (37.8%) of 75 GCs were CIMP-positive. DNA methylation of each of the 12 genes was observed more frequently in the high-methylation group than in the low methylation group. Methylation of 6 specific genes occurred more frequently in CIMP-positive GC than in CIMP-negative GC. Methylation of the remaining 6 genes was not correlated with CIMP-status. High methylation was found more frequently in Stage III/IV GC (26 of 40 cases, 65.0%) than in Stage I/II GC (13 of 35 cases, 37.1%, P = 0.029).CONCLUSIONS.These findings indicate that GCs with higher numbers of methylated genes have more distinct DNA methylation profiles than the originally defined CIMP-positive GCs. DNA methylation of tumor-related genes accumulates in conjunction with tumor progression. PMID- 16475211 TI - Cisplatin-induced antitumor activity is potentiated by the soy isoflavone genistein in BxPC-3 pancreatic tumor xenografts. AB - BACKGROUND: The activation of nuclear factor kappaB (NF-kappaB) contributes to drug resistance in pancreatic carcinoma. The authors previously showed that the soy isoflavone genistein down-regulates the activation of NF-kappaB in many carcinoma cell lines in vitro. In the current study, they focused their investigation on testing whether the inactivation of NF-kappaB by genistein could enhance cisplatin-induced cell growth inhibition and apoptosis in BxPC-3 cells in vitro and antitumor activity of cisplatin in vivo. METHODS: BxPC-3 cells were preexposed to 25 microM genistein for 24 hours and then exposed to cisplatin (0.5 microM) for an additional 72 hours. A cell growth inhibition assay, an apoptosis assay, and an NF-kappaB electrophoretic mobility shift assay were conducted. For the in vivo study, a xenograft model of BxPC-3 cells in severe combined immunodeficient mice was used. Genistein was given at a dose of 800 microg/kg orally for 5 days, cisplatin was given at a dose of 9 mg/kg as an intraperitoneal bolus, and another group of mice received both cisplatin and genistein (given on Day 1 concurrently followed by genistein for 4 days). RESULTS: The combination of 25 microM genistein with 0.5 microM cisplatin resulted in significantly greater growth inhibition (P < 0.01) and more apoptosis in BxPC-3 cells compared with either agent alone. Preexposure of BxPC-3 cells to genistein abrogated cisplatin induced activation of NF-kappaB, which appeared to be consistent with the authors' hypothesis. The authors also demonstrated for the first time that the in vivo effect of genistein enhanced the antitumor activity of cisplatin. The tumor weight for the control, genistein, cisplatin, and combined genistein and cisplatin mice was 940 mg, 762 mg, 261 mg, and 108 mg, respectively. Most important, for the first time, the authors observed that the DNA-binding activity of NF-kappaB was inactivated in genistein-treated animal tumors, whereas cisplatin significantly induced NF-kappaB DNA binding activity, and this was completely abrogated in genistein-pretreated tumors that were exposed to cisplatin, consistent with the in vitro data. CONCLUSIONS: Overall, the current results were consistent with the authors' hypothesis and suggested that pretreatment of pancreatic carcinoma cells with genistein down-regulates NF kappaB activity and contributes toward enhancing the apoptosis-inducing effect of cisplatin, leading to greater antitumor activity in vivo. PMID- 16475212 TI - Outcome of patients with fibrolamellar hepatocellular carcinoma. AB - BACKGROUND: Fibrolamellar hepatocellular carcinoma (FL-HCC) is a rare variant of hepatocellular carcinoma, has distinct pathologic features, and typically occurs in young patients without underlying hepatitis or cirrhosis. METHODS: Forty-one patients with the pathologic diagnosis of FL-HCC evaluated at our institution between 1986 and 2003 were identified from a prospective database. RESULTS: Median age of all patients was 27 years. None of these patients had underlying hepatitis or cirrhosis, and only 3 (7%) patients had an alpha-fetoprotein level > 200 ng/mL. Twenty-eight patients with primary disease underwent complete gross resection, and 13 patients were unresectable. In patients treated with resection, median tumor size was 9 cm (range, 3-17), 9 (36%) had vascular invasion, and 14 (50%) had lymph node metastases. There were no perioperative deaths. With a median follow-up of 34 months, 5-year overall survival for resected patients was 76%. However, 5-year recurrence-free survival was only 18%, and of the 9 resected patients with more than 5 years of follow-up, 7 had recurrences. Lymph node metastasis was the only significant negative prognostic factor. Seventeen (61%) patients underwent a second operation for recurrent disease. Median survival for unresected patients with FL-HCC was only 12 months, and no patient survived beyond 5 years. CONCLUSIONS: FL-HCC occurs in a distinctly different population of patients than common HCC, and patients with FL-HCC generally fare better after complete resection. These tumors have a relatively indolent tumor biology, and late recurrences are common. Repeat resections for recurrence should be considered given the lack of other effective treatment options. PMID- 16475213 TI - A Phase II study of gemcitabine and cisplatin in advanced biliary tract cancer. AB - BACKGROUND: The authors performed a Phase II study of combination chemotherapy with gemcitabine and cisplatin in patients with inoperable biliary tract cancer to evaluate efficacy and toxicity of this combination. In addition, the correlation between the CA 19-9 response and clinical outcome was analyzed. METHODS: The eligibility criteria for this study were 1) histologically or cytologically confirmed inoperable biliary tract cancer in patients with metastatic or recurrent disease; 2) age between 18 and 70 years; 3) at least 1 measurable lesion according to Response Evaluation Criteria in Solid Tumors (RECIST) criteria; 4) an Eastern Cooperative Oncology Group (ECOG) performance status < or = 2; 5) a life expectancy of at least 3 mos; and 6) adequate bone marrow, hepatic, and renal function. The patients received gemcitabine (1250 mg/m(2), Days 1 and 8) and cisplatin (60 mg/m(2), Day 1) every 3 weeks. Tumor response was assessed by RECIST criteria every 2 cycles of chemotherapy. Treatment was continued until progression of disease was documented. RESULTS: Twenty-nine patients were enrolled. The median age of these patients was 52 years (range, 37 to 69 yrs), and the median ECOG performance status was 1. No complete response was observed, and 10 of 29 patients had partial responses. The overall response rate was 34.5% (95% confidence interval [CI], 17.9-54.3) for the intent to-treat analysis. Stable disease was observed in 4 (13.8%) patients and progressive disease in 13 (44.8%) patients. The median follow-up time was 10.0 months (95% CI, 7.2-12.8). The median time to progression (TTP) was 3.0 months (95% CI, 2.12-3.88), and the median overall survival was 11 months (95% CI, 5.49 16.5). Although these results showed a better response rate (57.1 % vs. 27.3%) and survival (12 vs. 10 mos) in patients with a decline in CA 19-9 of at least 25%, these data were statistically not significant. In addition, there was a significant positive correlation between the increment in CA 19-9 values and tumor progression as determined with RECIST criteria (r = 0.96, P < 0.01). However, there was no definite correlation between the CA 19-9 response and the response according to RECIST criteria (P = 0.087). National Cancer Institute (NCI) common toxicity criteria (CTC) Grade 3 or 4 hematologic toxicities included neutropenia in 4 (14%) patients and anemia in 1 (3%) patient. Two of 4 patients with Grade 3 or 4 neutropenia had febrile episodes (7%) and required hospital admissions. NCI-CTC Grade 3 or 4 nonhematologic toxicity included nausea in 1 (3%) patient. There were no treatment-related deaths. CONCLUSION: The combination chemotherapy with gemcitabine and cisplatin in inoperable biliary tract cancer was tolerable for most patients and showed modest response rates. The role of CA 19-9 monitoring as a surrogate biomarker in patients with BTC treated with gemcitabine chemotherapy should be further investigated. PMID- 16475214 TI - Structural and magnetic properties of Mn(III) and Cu(II) tetranuclear azido polyoxometalate complexes: multifrequency high-field EPR spectroscopy of Cu4 clusters with S = 1 and S = 2 ground states. AB - Two new azido-bridged polyoxometalate compounds were synthesized in acetonitrile/methanol media and their molecular structures have been determined by X-ray crystallography. The [[(gamma-SiW10O36)Mn2(OH)2(N3)(0.5)(H2O)(0.5)]2(mu 1,3-N3)](10-) (1 a) tetranuclear Mn(III) complex, in which an end-to-end N3- ligand acts as a linker between two [(gamma-SiW10O36)Mn2(OH)2]4- units, represents the first manganese-azido polyoxometalate. The magnetic properties have been studied considering the spin Hamiltonian H = -J1(S1S2+S1*S2*) J2(S1S1*), showing that antiferromagnetic interactions between the paramagnetic centers (g = 1.98) occur both through the di-(mu-OH) bridge (J1 = -25.5 cm(-1)) and the mu-1,3-azido bridge (J2 = -19.6 cm(-1)). The [(gamma-SiW10O36)2Cu4(mu 1,1,1-N3)2(mu-1,1-N3)2]12- (2 a) tetranuclear Cu(II) complex consists of two [gamma-SiW10O36Cu2(N3)2]6- subunits connected through the two mu-1,1,1-azido ligands, the four paramagnetic centers forming a lozenge. The magnetic susceptibility data have been fitted. This reveals ferromagnetic interactions between the four Cu(II) centers, leading to an S=2 ground state (H = J1(S1S2+S1*S2*)-J2(S2S2*), J1 = +294.5 cm(-1), J2 = +1.6 cm(-1), g = 2.085). The ferromagnetic coupling between the Cu(II) centers in each subunit is the strongest ever observed either in a polyoxometalate compound or in a diazido bridged Cu(II) complex. Considering complex 2 a and the previously reported basal basal di-(mu-1,1-N3)-bridged Cu(II) complexes in which the metallic centers are not connected by other magnetically coupling ligands, the linear correlation J1 = 2639.5-24.95*theta(av) between the theta(av) bridging angle and the J1 coupling parameter has been proposed. The electronic structure of complex 2 a has also been investigated by using multifrequency high-field electron paramagnetic resonance (HF-EPR) spectroscopy between 95 and 285 GHz. The spin Hamiltonian parameters of the S = 2 ground state (D = -0.135(2) cm(-1), E = -0.003(2) cm(-1), g(x) = 2.290(5), g(y) = 2.135(10), g(z) = 2.158(5)) as well as of the first excited spin state S = 1 (D = -0.960(4) cm(-1), E = -0.080(5) cm(-1), g(x) = 2.042(5), g(y) = 2.335(5), g(z) = 2.095(5)) have been determined, since the energy gap between these two spin states is very small (1.6 cm(-1)). PMID- 16475215 TI - Upper susceptibility threshold limits with confidence intervals: a method to identify normal and abnormal population values for laboratory toxicological parameters, based on acetylcholinesterase activities in sea lice. AB - The interpretation and importance of comparing field values of susceptibility to pesticides with a laboratory reference strain that might bear little resemblance to the actual situation in the field are problematic and a continuing subject of debate. In this paper a procedure for defining a 'normal sensitive' population from a field study of 383 individuals to provide a basis for analysing and interpreting in vitro results is described and examined. Instead of using only the 95th percentile, the upper and lower confidence limits for the 95th percentile were also compared to select the best estimation of the limit for the normal material. A field population constrained by the upper confidence limit for the 95th percentile provides appropriate descriptions of the normal material in this study. This approach should prove useful in studies of pesticide resistance in field populations. PMID- 16475216 TI - Influence of local and systemic CTLA4Ig gene transfer on corneal allograft survival. AB - BACKGROUND: To analyse the effects of local (ex vivo) or systemic (in vivo) administration of adenovirus type 5 encoding CTLA4Ig (AdCTLA4Ig) on its influence to prolong corneal allograft survival and to study the underlying mechanisms. METHODS: A MHC class I/II mismatched rat corneal transplant model was used. Recipients were randomly assigned to receive ex vivo gene-modified corneas expressing either CTLA4Ig, CTLA4Ig/IL-10 or a single intraperitoneal (i.p.) injection (1.0 x 10(9) or 1.0 x 10(10) infectious particles) of AdCTLA4Ig 1 day before transplantation and graft survival was analysed. The immunoregulatory effect of this treatment was examined by analysing intra-graft cytokine mRNA expression pattern at day 12 post-transplant. The anti-adenovirus immunity also was investigated. RESULTS: Ex vivo gene transfer resulted in a modest but significant prolongation of graft survival (p = 0.0036 compared to no treatment). In contrast, systemic gene therapy (1.0 x 10(9) or 1.0 x 10(10) infectious particles) significantly prolonged graft survival (p = 0.0007 and 0.0001, respectively, compared to no treatment). Systemic (1.0 x 10(10) infectious particles) therapy resulted in frequent indefinite survival of allogeneic grafts which was not observed in the other therapeutic regimens. Moreover, systemic therapy prevented the intra-graft accumulation and activation of T cells and resulted in a reduced mRNA expression of both TH1 and TH2 cytokines. The generation of anti-adenovirus antibodies was also efficiently inhibited. CONCLUSIONS: CTLA4Ig gene therapy is a successful strategy for the prevention of allogeneic graft rejection in corneal transplantation. Our work has further elucidated the mechanisms of corneal allograft rejection which may lead to novel therapeutic strategies. PMID- 16475218 TI - Phytogrowth- and photosynthesis-inhibiting properties of nostoclide analogues. AB - Six nostoclide analogues were synthesised from 3-benzyl-2(5H)-furanone in one step, with yields ranging from 10 to 71%, and subjected to several biological assays. The two most active of these, 5d and 5e, were shown to be phytogrowth inhibitors of the radicle of Lolium multiflorum Lam, while enhancing the root growth of Physalis ixocarpa Brot. Both compounds inhibited electron flow (basal, phosphorylating and uncoupled) from water to methylviologen (MV); both acted as Hill reaction inhibitors, since the synthesis of ATP was prevented. The uncoupled electron transport from photosystem II (PSII) (water to 2,6-dichlorophenol indophenol (DPIP)) and photosystem I (PSI) (2,6-dichlorophenol-indophenol reduced (DPIPred) to MV) was inhibited with 500 microM of 5d by 22 and 14% respectively. In addition, 400 microM of 5d inhibited PSI (from tetramethyl-p-benzohydroquinone (TMQH(2)) to MV) by 40%. Thus 5d inhibited electron transport at the b(6)f complex. Finally, 500 microM of 5e inhibited electron flow (basal and phosphorylating) by 25%, and 300 microM of 5e enhanced light-activated membrane bound Mg(2+)-ATPase by 66%. Thus 5e behaved as a weak Hill reaction inhibitor and an uncoupler. In general, the phytotoxicity of the synthetic lactones was only weakly related to inhibition of photosynthesis. PMID- 16475217 TI - Development and characterization of a synthetic promoter for selective expression in proliferating endothelial cells. AB - BACKGROUND: Systemic administration of non-viral gene therapy provides better access to tumors than local administration. Development of a promoter that restricts expression of cytotoxic proteins to the tumor vasculature will increase the safety of the system by minimizing expression in the non-dividing endothelial cells of the vasculature of non-target tissues. METHODS: Cell cycle promoters were tested for selective expression in dividing cells vs. non-dividing cells in vitro and promoter strength was compared to the cytomegalovirus (CMV) promoter. Successful promoter candidates were tested in vivo using two proliferating endothelium mouse models. Ovarectomized mice were injected with estradiol prior to lipoplex administration and expression levels were measured in the lungs and uterus 4 days after administration. The second model was a subcutaneous tumor model and expression levels were measured in the lungs and tumors. For both animal models, expression levels from the proliferating endothelium promoter were compared to that obtained from a CMV promoter. RESULTS: The results showed that the Cdc6 promoter yielded higher expression in proliferating vs. non proliferating cells. Secondly, promoter strength could be selectively increased in endothelial cells by the addition of a multimerized endothelin enhancer (ET) to the Cdc6 promoter. Thirdly, comparison of expression levels in the lungs vs. uterus in the ovarectomized mouse model and lungs vs. tumor in the mouse tumor model showed expression was much higher in the uterus and the tumor than in the lungs for the ET/Cdc6 promoter, and expression levels were comparable to that of the CMV promoter in the hypervascularized tissues. CONCLUSIONS: These results demonstrate that the combination of the endothelin enhancer with the Cdc6 promoter yields selective expression in proliferating endothelium and can be used to express cytotoxic proteins to treat vascularized tumors. PMID- 16475219 TI - Acaricidal activity against Panonychus citri of a ginkgolic acid from the external seed coat of Ginkgo biloba. AB - An acaricidal substance extracted from the external seed coat of Ginkgo biloba L. was identified by UV (ultraviolet), IR (infrared), EI-MS (electron impact ion source mass spectrometry), (1)H NMR (nuclear magnetic resonance) and (13)C NMR as 6-[(Z)-10-heptadecenyl]-2-hydroxybenzoic acid (compound 1). Laboratory bioassay on citrus red mite, Panonychus citri (Mcg), showed that compound 1 possessed the following properties. (i) Powerful contact toxicity with an LC(50) of 5.2 mg litre(-1) after 24 h that was similar to that of pyridaben (LC(50) = 3.4 mg litre(-1)) and significantly superior to that of omethoate (LC(50) = 122 mg litre(-1)). Furthermore, its LC(90) was 13.4 mg litre(-1) after 24 h, which is significantly superior to both pyridaben (LC(90) = 69.6 mg litre(-1)) and omethoate (LC(90) = 453 mg litre(-1)). (ii) Quick-acting acaricidal activity. At identical concentrations, compound 1 was much faster-acting than pyridaben or omethoate. (iii) Compound 1 had strong corrosive action on the cuticle of P. citri but no phytotoxicity to plants. PMID- 16475220 TI - Baylis-Hillman adducts with molluscicidal activity against Biomphalaria glabrata. AB - The molluscicidal activities of ten Baylis-Hillman adducts against Biomphalaria glabrata (Say) snails, the intermediate host of schistosomiasis, have been determined. Nine of these compounds showed significant molluscicidal activity against B. glabrata, falling below the threshold of 100 microg ml(-1) set for potential molluscicidal activity by the World Health Organisation. Among these compounds, 3-hydroxy-2-methylene-3-(4-nitrophenyl)propanenitrile had the highest activity, with LC(50) = 6.64 microg ml(-1). PMID- 16475221 TI - Influence of Bacillus spp. isolated from maize agroecosystem on growth and aflatoxin B(1) production by Aspergillus section Flavi. AB - A total of 59 bacteria of the Bacillus genus were isolated from different components of a maize agroecosystem and their antifungal activity against Aspergillus section Flavi was evaluated. Thirty-three and 46% of these bacteria were able to inhibit Aspergillus flavus Link and A. parasiticus Speare respectively at water activity (a(w)) 0.982; however, when a(w) was 0.955, these percentages were decreased and only three isolates were able to inhibit Aspergillus section Flavi. The majority of bacilli acted as contact antagonists, while a small number of isolates were able to form inhibition zones. In maize meal extract agar, Aspergillus section Flavi growth rate and aflatoxin B(1) (AFB(1)) production were significantly reduced when these strains were paired at a(w) 0.982 with bacilli at all inoculum levels studied. However, two bacilli isolated were able to reduce growth rate and aflatoxin production when a(w) was 0.955. Lag phase increase followed the same general pattern as growth rate reduction. When Aspergillus section Flavi was grown in sterile maize in the presence of three Bacillus strains at a(w) 0.982, the reduction in count (colony forming units (cfu) g(-1) maize) was less than 30%, except when Aspergillus section Flavi grew with Bacillus amyloliquefaciens UNRCLR. However, levels of detectable AFB(1) were significantly reduced in these interactions at a(w) 0.982. PMID- 16475222 TI - Residues of diflubenzuron on horse chestnut (Aesculus hippocastanum) leaves and their efficacy against the horse chestnut leafminer, Cameraria ohridella. AB - Residues of the insect growth regulator diflubenzuron were quantified on horse chestnut (Aesculus hippocastanum L.) leaves treated with a diflubenzuron 480 g litre(-1) SC, Dimilin. To analyse the samples, an analytical procedure was developed involving a simple extraction step followed by high-performance liquid chromatography on an octadecyl-modified silica column with methanol + 0.01 M ammonium acetate mobile phase. The results showed diflubenzuron to be highly stable on horse chestnut leaves; more than 4 months (127 days) after application, 38% (on average) of the insecticide still remained on/in the leaves. The data confirmed biological observations showing diflubenzuron's long-term efficacy against the horse chestnut leafminer, Cameraria ohridella Deschka and Dimic, which is the most important pest of the horse chestnut in Europe. The hypothesis of possible penetration of diflubenzuron into the leaf mass is explored and discussed. PMID- 16475223 TI - Fitness costs of laboratory-selected imidacloprid resistance in the brown planthopper, Nilaparvata lugens Stal. AB - Imidacloprid has been used as a key insecticide to control the brown planthopper, Nilaparvata lugens Stal, for several years, but no obvious resistance has been identified in field populations as yet. To evaluate the risk, a field population was collected and selected with imidacloprid in the laboratory. After 37 generation selection a strain with 250-fold resistance had been successfully achieved. Fitness analysis by constructing life tables demonstrated that resistant hoppers had obvious disadvantages in their reproduction. The fitness of highly resistant hoppers had decreased dramatically (0.169 and 0.104) to only one fifth to one-tenth of that of the susceptible strain. Hence it was concluded that the brown planthopper had the potential to develop high resistance to imidacloprid but that the lower fitness of resistant hoppers could result in a quick recovery of sensitivity when the population did not come into contact with imidacloprid. This means that a reasonable resistance management programme with less imidacloprid use may efficiently delay or even stop resistance development. PMID- 16475224 TI - Shaping amorphous calcium carbonate films into 2D model substrates for bone cell culture. PMID- 16475225 TI - Postzygotic isochromosome formation as a cause for false-negative results from chorionic villus chromosome examinations. AB - OBJECTIVE: To investigate the origin and mechanisms of formation of isochromosomes 13q and 21q in instances where prenatal chromosome examination revealed a normal karyotype while postnatal chromosome examination from blood showed translocation trisomy 13 and 21. METHODS: G and/or Q-banded chromosome examinations from CVS cultures and lymphocyte chromosome examinations from two newborns. Microsatellite marker analysis of DNA from the probands and their parents. Prenatal ultrasonic examinations of the fetuses and postnatal clinical examinations of the probands. RESULTS: Short and long-term CVS examinations from two fetuses revealed normal karyotypes. Lymphocyte karyotypes of the newborns showed the karyotype 46,XY,i(21)(q10) in the first case and 46,XY,i(13)(q10) in the second. The isochromosomes 21q and 13q were shown, by microsatellite marker analysis of the patients and their parents, to be of maternal and paternal origin, respectively. CONCLUSION: Postzygotic isochromosome formation is one of the possible mechanisms that may lead to false-negative results of chorionic villus chromosome examinations, even if both short-term and long-term cultures are performed and give normal results. PMID- 16475226 TI - Prenatal diagnosis of citrullinemia and argininosuccinic aciduria: evidence for a transmission ratio distortion in citrullinemia. AB - BACKGROUND: In the course of 25 years, we have experienced a high rate of affected fetuses in the prenatal diagnosis of citrullinemia. METHODS AND RESULTS: Ninety-one pregnancies at 1 in 4 risk were tested; 36 were diagnosed as affected (39.5%; P = 0.0015). The high rate of positive diagnoses was found both after chorionic villus sampling (24/68 = 35.3%) and amniocentesis (12/23 = 52.2%) despite the completely different and independent techniques used. Using exactly the same (indirect) enzyme assay for argininosuccinic aciduria on chorionic villi and a similar method on amniotic fluid, the expected rate of affected fetuses was found: 13/53 = 24.5%. Technical and genetic causes for the unexpected results were excluded by confirmatory studies performed on independent fetal material, which was available for 27 of the 36 fetuses affected with citrullinemia. Biochemical confirmation was obtained in the 27 cases, whereas in 18 fetuses homozygosity or compound heterozygosity for disease-causing mutations were retrospectively demonstrated in the stored fetal cells. CONCLUSION: The results suggest the occurrence of preferential transmission of the mutant allele. An explanation for this phenomenon may be found in a protective role of argininosuccinic acid synthetase deficiency in mutant sperm cells against the possibly detrimental or apoptotic effect of nitric oxide produced normally from arginine by nitric oxide synthase. PMID- 16475227 TI - Attitudes towards prenatal diagnosis and termination of pregnancy for thalassaemia in pregnant Pakistani women in the North of England. AB - OBJECTIVES: Most births of children affected with beta-thalassaemia major in the United Kingdom are to parents of Pakistani origin. A popular explanation for this is that Pakistanis decline termination of pregnancy on religious grounds. However, various factors influence people's attitudes towards prenatal diagnosis and termination of pregnancy, which have not been investigated in a UK Pakistani sample. This study is aimed at exploring the attitudes of pregnant Pakistani women towards prenatal diagnosis and termination of pregnancy for beta thalassaemia major in the North of England. METHODS: Forty-three pregnant women tested for thalassaemia carrier status were interviewed following receipt of their test results. Interviews were analysed using the grounded theory approach. RESULTS: Findings showed: (1) women's awareness of and attitudes towards prenatal diagnosis; (2) the relationship between attitudes towards prenatal diagnosis and termination of an affected foetus; (3) the relationship between attitudes towards termination of pregnancy and religious beliefs, perceptions of severity of the condition, influence of significant others, and (4) the impact of gestational age at the time of the offer of termination of pregnancy. CONCLUSIONS: Pakistani women's attitudes towards prenatal diagnosis and termination of pregnancy are influenced by various factors, and therefore their religion should not be taken as a proxy for their attitudes either for or against termination of pregnancy. PMID- 16475228 TI - Oriented attachment mechanism in anisotropic nanocrystals: a "polymerization" approach. AB - The kinetic model of stepwise polymerization is revisited, with some adaptations for its application to the kinetics of oriented attachment of nanoparticles in colloidal suspensions, which results in the formation of anisotropic particles. A comparison with experimental data reported in the literature shows good agreement with the model and supports comparisons with other systems. PMID- 16475229 TI - An automatable screen for the rapid identification of proteins amenable to refolding. AB - Insoluble expression of heterologous proteins in Escherichia coli is a major bottleneck of many structural genomics and high-throughput protein biochemistry projects. Many of these proteins may be amenable to refolding, but their identification is hampered by a lack of high-throughput methods. We have developed a matrix-assisted refolding approach in which correctly folded proteins are distinguished from misfolded proteins by their elution from affinity resin under non-denaturing conditions. Misfolded proteins remain adhered to the resin, presumably via hydrophobic interactions. The assay can be applied to insoluble proteins on an individual basis but is particularly well suited for high throughput applications because it is rapid, automatable and has no rigorous sample preparation requirements. The efficacy of the screen is demonstrated on small-scale expression samples for 15 proteins. Refolding is then validated by large-scale expressions using SEC and circular dichroism. PMID- 16475230 TI - Proteomic analysis of membrane microdomains derived from both failing and non failing human hearts. AB - Eukaryotic cells plasma membranes are organized into microdomains of specialized function such as lipid rafts and caveolae, with a specific lipid composition highly enriched in cholesterol and glycosphingolipids. In addition to their role in regulating signal transduction, multiple functions have been proposed, such as anchorage of receptors, trafficking of cholesterol, and regulation of permeability. However, an extensive understanding of their protein composition in human heart, both in failing and non-failing conditions, is not yet available. Membrane microdomains were isolated from left ventricular tissue of both failing (n = 15) and non-failing (n = 15) human hearts. Protein composition and differential protein expression was explored by comparing series of 2-D maps and subsequent identification by LC-MS/MS analysis. Data indicated that heart membrane microdomains are enriched in chaperones, cytoskeletal-associated proteins, enzymes and protein involved in signal transduction pathway. In addition, differential protein expression profile revealed that 30 proteins were specifically up- or down-regulated in human heart failure membrane microdomains. This study resulted in the identification of human heart membrane microdomain protein composition, which was not previously available. Moreover, it allowed the identification of multiple proteins whose expression is altered in heart failure, thus opening new perspectives to determine which role they may play in this disease. PMID- 16475231 TI - Analysis of chicken serum proteome and differential protein expression during development in single-comb White Leghorn hens. AB - Serum is believed to harbor thousands of distinct proteins that are either actively secreted or leak from various blood cells or tissues. Exploring protein composition in serum may accelerate the discovery of novel protein biomarkers for specific economic traits in livestock species. This study analyzed serum protein composition to establish a 2-DE reference map, and monitored protein dynamics of single-comb White Leghorn hens at 8, 19 and 23 weeks after hatching. A total of 119 CBB-stained and 315 silver-stained serum protein spots were analyzed by MALDI TOF MS. Of these, 98 CBB-stained and 94 silver-stained protein spots were significantly matched to existing chicken proteins. The identified spots represented 30 distinctive proteins in the serum of laying hens. To compare protein expression during development, expression levels of 47 protein spots were quantified by relative spot volume with Melanie 3 software. Ten protein spots increased and 3 protein spots decreased as hen age increased. Previous research has suggested that some of these proteins play critical roles in egg production. The differentially expressed proteins with unknown identities will be valuable candidates for further explorations of their roles in egg production of laying hens. PMID- 16475232 TI - Comparison of the Paul ion trap to the linear ion trap for use in global proteomics. AB - A critical evaluation of the performance of a 2-D linear ion trap (IT) instrument to two 3-D quadrupole IT instruments with emphasis on identification of rat serum proteins by bottom-up LC-MS/MS is presented. The speed and sensitivity of each of the instruments were investigated, and the effects that each of these have on the bottom-up proteomics identification approach are discussed. PMID- 16475233 TI - 2-DE proteome analysis of a proliferating and differentiating human neuronal stem cell line (ReNcell VM). AB - The proteome of a proliferating human stem cell line was analyzed and then utilized to detect stem cell differentiation-associated changes in the protein profile. The analysis was conducted with a stable human fetal midbrain stem cell line (ReNcell VM) that displays the properties of a neural stem cell. Therefore, acquisition of proteomic data should be representative of cultured human neural stem cells (hNSCs) in general. Here we present a 2-DE protein-map of this cell line with annotations of 402 spots representing 318 unique proteins identified by MS. The subsequent proteome profiling of differentiating cells of this stem cell line at days 0, 4 and 7 of differentiation revealed changes in the expression of 49 identified spots that could be annotated to 45 distinct proteins. This differentiation-associated expression pattern was validated by Western blot analysis for transgelin-2, proliferating cell nuclear antigen, as well as peroxiredoxin 1 and 4. The group of regulated proteins also included NudC, ubiquilin-1, STRAP, stress-70 protein, creatine kinase B, glial fibrillary acidic protein and vimentin. Our results reflect the large rearrangement of the proteome during the differentiation process of the stem cells to terminally differentiated neurons and offer the possibility for further characterization of specific targets driving the stem cell differentiation. PMID- 16475234 TI - Prenatal diagnosis of achondroplasia presenting with multiple-suture synostosis: a novel association. AB - OBJECTIVE: We report an atypical case of a fetus presenting with a combined achondroplasia and multiple craniosynostosis phenotype. METHODS: Sonographic monitoring in conjunction with molecular genetic analysis was performed in a 32 gestational weeks fetus. RESULTS: Sonographic findings were consistent with a diagnosis of achondroplasia associated with multiple-suture synostosis. The most common G380R FGFR3 achondroplasia mutation was detected. CONCLUSION: The most common achondroplasia mutation should be considered for prenatal DNA testing in cases with ultrasound findings of achondroplasia and multiple-suture synostosis. This is crucial for the genetic counselling and perinatal management of the fetus. PMID- 16475235 TI - A novel heterozygous missense mutation 377T > C (V126A) of TGIF gene in a family segregated with holoprosencephaly and moyamoya disease. AB - OBJECTIVES: To identify whether any mutations of candidate genes including SHH, ZIC2, SIX3, and TGIF exist in a Taiwanese family segregated with holoprosencephaly (HPE) and moyamoya disease. METHODS: Genotypes of the candidate genes SHH, ZIC2, SIX3, and TGIF were determined in the family members who were available for analysis by sequencing. In addition, genomic regions of another 50 unrelated Taiwanese (100 chromosomes) were studied to verify whether the nucleotide changes we found were mutations or polymorphisms. RESULTS: A novel missense mutation 377T > C and two polymorphisms (420A > G and 487C > T) in the TGIF gene were identified. No mutations in SHH, ZIC2 and SIX3 were found. The mother of the three HPE fetuses was found to be afflicted with moyamoya disease. A brief review of the mutations as well as polymorphisms reported in the TGIF gene up to 2005 is given. CONCLUSION: Molecular diagnosis can help genetic counseling in HPE, which is a heterogeneous disorder with its phenotypic and genotypic spectrum highly widened and variable. The possible association between TGIF mutation and moyamoya disease noted in our study also appeared to be novel. PMID- 16475236 TI - The use of an alternative food source (red clover) as a means of reducing slug pest damage to winter wheat: towards field implementation. AB - Slugs are major pests of many crops in the UK, including winter wheat, yet current methods of control are often unreliable. The aim of this study was to investigate three issues key to the successful field implementation of a control strategy that uses red clover as an alternative food source to reduce the amount of damage caused to winter wheat by the field slug, Deroceras reticulatum (Muller). A series of three experiments was designed to assess this aim. Firstly, under laboratory conditions, red clover was consumed in greater quantities than wheat, even when wheat was presented as a novel food. Secondly, red clover had no significant effects on the emergence and early growth of wheat in a polytunnel experiment. Both these results are crucial to the successful implementation of a strategy that uses red clover as an alternative food source. Lastly, the results of a field experiment were consistent with the results of the polytunnel experiment, in that red clover did not significantly affect wheat emergence. However, plots in which red clover was left to grow until the time of wheat harvest resulted in significantly lower (43%) wheat yields than plots without red clover. These results suggest that red clover must be removed from the field after the wheat has passed its vulnerable seedling stage. Recommendations for the potential use of red clover as an alternative food source for reducing damage to winter wheat in field conditions are discussed and opportunities for further work are suggested. PMID- 16475237 TI - Substituted alpha-(phenylhydrazono)phenylacetonitrile derivatives. Part 2: Synthesis and biological activity of pro-pesticides. AB - Pro-pesticides of alpha-(2,6-dichloro-4-trifluoromethylphenylhydrazono)-4 nitrophenylacetonitrile have been prepared and tested against mite and insect pests. Variations in potency and spectrum were observed depending on the choice of cleavable pro-moiety. Cleavage of the pro-moiety was demonstrated in one case by measuring the rate of increase in the uncoupling activity using a mitochondrial preparation. Irradiation experiments have demonstrated a rapid isomerisation of the planar Z isomer to the E isomer, which is reversible. PMID- 16475238 TI - Fumigant distribution in forest nursery soils under water seal and plastic film after application of dazomet, metam-sodium and chloropicrin. AB - Adequate concentration, exposure time and distribution uniformity of activated fumigant gases are prerequisites for successful soil fumigation. Field experiments were conducted to evaluate gas phase distributions of methyl isothiocyanate (MITC) and chloropicrin (CP) in two forest-tree nurseries. Concentrations of MITC and CP in soil air were measured from replicated microplots that received dazomet, metam-sodium and CP. Half of the plots were covered with high-density polyethylene tarp immediately after fumigation; the other half were not covered but received daily sprinkler irrigation for 1 week to create and maintain a water seal. The magnitude of MITC concentrations was similar between nurseries for metam-sodium in both tarp and water seal treatments and for dazomet in the tarp treatment. Consistently greater MITC and CP concentrations were found in the upper 30 cm of soil in the tarped plots compared with the water-sealed plots. Despite potential environmental and economic benefits with the water seal method, tarp covers were more reliable for achieving and maintaining higher MITC and CP concentrations and less prone to variations due to irrigation/rain, soil bulk density and other environmental conditions. PMID- 16475239 TI - In vitro selection of an effective fungicide against Armillaria mellea and control of white root rot of grapevine in the field. AB - Armillaria mellea (Vahl ex Fr) Kummer is an aggressive pathogen which causes white root rot in a wide range of hosts. Most chemicals tested so far against Armillaria, both in vitro and in the field, have not been effective in reducing fungal growth and/or preventing plant decline and mortality. In the present work the effects of four DMI (sterol demethylation inhibitor) fungicides, cyproconazole, hexaconazole, propiconazole and tetraconazole, and another six downwardly mobile systemic chemicals, azoxystrobin, cubiet (copper bis(ethoxy dihydroxy-diethylamino)sulfate), fosetyl-Al, potassium phosphite, sodium tetrathiocarbonate (STTC) and 2-(thiocyanomethylthio)benzothiazole (TCMTB), on the mycelial growth of A. mellea were compared and evaluated; the product yielding the best results in in vitro experiments was selected to determine its efficacy in preventing decline and mortality of grapevines in the field. Best results on in vitro fungal growth reduction were obtained with the four azoles tested, in particular with cyproconazole and hexaconazole, achieving 67-72% mycelial growth inhibition at the lowest dose. Results obtained in the field showed that a dose of 50 mg AI litre(-1) of cyproconazole once or twice a year was efficient in controlling the disease even in vines seriously affected by the pathogen. However, further research is required to study minimum effective doses, residual effects and the convenience of the application of annual dressings in damaged vineyards, so as to gradually reduce the pathogen inoculum potential in soil and control the disease while reducing chemical residues in the plant and preventing development of fungal resistance. PMID- 16475240 TI - Impact of fungal drug transporters on fungicide sensitivity, multidrug resistance and virulence. AB - Drug transporters are membrane proteins that provide protection for organisms against natural toxic products and fungicides. In plant pathogens, drug transporters function in baseline sensitivity to fungicides, multidrug resistance (MDR) and virulence on host plants. This paper describes drug transporters of the filamentous fungi Aspergillus nidulans (Eidam) Winter, Botrytis cinerea Pers and Mycosphaerella graminicola (Fuckel) Schroter that function in fungicide sensitivity and resistance. The fungi possess ATP-binding cassette (ABC) drug transporters that mediate MDR to fungicides in laboratory mutants. Similar mutants are not pronounced in field resistance to most classes of fungicide but may play a role in resistance to azoles. MDR may also explain historical cases of resistance to aromatic hydrocarbon fungicides and dodine. In clinical situations, MDR development in Candida albicans (Robin) Berkhout mediated by ABC transporters in patients suffering from candidiasis is common after prolonged treatment with azoles. Factors that can explain this striking difference between agricultural and clinical situations are discussed. Attention is also paid to the risk of MDR development in plant pathogens in the future. Finally, the paper describes the impact of fungal drug transporters on drug discovery. PMID- 16475241 TI - Substituted alpha-(phenylhydrazono)phenylacetonitrile derivatives. Part 1: A new class of uncoupler of oxidative phosphorylation. AB - Substituted alpha-(phenylhydrazono)phenylacetonitrile derivatives have been discovered which constitute a series of potent uncouplers of oxidative phosphorylation. Systematic variation of substituents on both benzene rings has clearly demonstrated the importance of steric congestion around the ionisation site and delocalisation of negative charge in the anionic form. Replacement of the cyano group by other electron-withdrawing groups leads to a dramatic decrease in uncoupling activity. The sub-nanomolar levels of uncoupling activity found in certain members indicate that these compounds are the most potent uncouplers yet reported. PMID- 16475243 TI - Efficient in toto targeted recombination in mouse liver by meganuclease-induced double-strand break. AB - BACKGROUND: Sequence-specific endonucleases with large recognition sites can cleave DNA in living cells, and, as a consequence, stimulate homologous recombination (HR) up to 10 000-fold. The recent development of artificial meganucleases with chosen specificities has provided the potential to target any chromosomal locus. Thus, they may represent a universal genome engineering tool and seem to be very promising for acute gene therapy. However, in toto applications depend on the ability to target somatic tissues as well as the proficiency of somatic cells to perform double-strand break (DSB)-induced HR. METHODS: In order to investigate DSB-induced HR in toto, we have designed transgenic mouse lines carrying a LagoZ gene interrupted by one I-SceI cleavage site surrounded by two direct repeats. The LagoZ gene can be rescued upon cleavage by I-SceI and HR between the two repeats in a process called single strand annealing. beta-Galactosidase activity is monitored in liver after tail vein injection of adenovirus expressing the meganuclease I-SceI. RESULTS: In toto staining revealed a strong dotted pattern in all animals injected with adenovirus expressing I-SceI. In contrast, no staining could be detected in the control. beta-Galactosidase activity in liver extract, tissue section staining, and PCR analysis confirmed the presence of the recombined LagoZ gene. CONCLUSIONS: We demonstrate for the first time that meganucleases can be successfully delivered in animal and induce targeted genomic recombination in mice liver in toto. These results are an essential step towards the use of designed meganucleases and show the high potential of this technology in the field of gene therapy. PMID- 16475242 TI - Imaging and therapy of malignant pleural mesothelioma using replication-competent herpes simplex viruses. AB - BACKGROUND: Malignant pleural mesothelioma (MPM) is an aggressive cancer that is refractory to current treatment modalities. Oncolytic herpes simplex viruses (HSV) used for gene therapy are genetically engineered, replication-competent viruses that selectively target tumor cells while sparing normal host tissue. The localized nature, the potential accessibility and the relative lack of distant metastasis make MPM a particularly suitable disease for oncolytic viral therapy. METHODS: The infectivity, selective replication, vector spread and cytotoxic ability of three oncolytic HSV: G207, NV1020 and NV1066, were tested against eleven pathological types of MPM cell lines including those that are resistant to radiation therapy, gemcitabine or cisplatin. The therapeutic efficacy and the effect on survival of NV1066 were confirmed in a murine MPM model. RESULTS: All three oncolytic HSV were highly effective against all the MPM cell lines tested. Even at very low concentrations of MOI 0.01 (MOI: multiplicity of viral infection, ratio of viral particles per cancer cell), HSV were highly effective against MPM cells that are resistant to radiation, gemcitabine and cisplatin. NV1066, an oncolytic HSV that expresses green fluorescent protein (GFP), was able to delineate the extent of the disease in a murine model of MPM due to selective infection and expression of GFP in tumor cells. Furthermore, NV1066 was able to reduce the tumor burden and prolong survival even when treatment was at an advanced stage of the disease. CONCLUSION: These findings support the continued investigation of oncolytic HSV as potential therapy for patients with therapy resistant MPM. PMID- 16475246 TI - HIV epidemic continues to make major inroads in sub-Saharan Africa. AIDS deaths estimated at 2.4 million. PMID- 16475245 TI - Alcohol advertising and alcohol consumption by adolescents. AB - This study investigates the effects of alcohol advertising on adolescent alcohol consumption. The theory of an industry response function and evidence from prior studies indicate the importance of maximizing the variance in advertising measures. Monitoring the Future (MTF) and National Longitudinal Survey of Youth 1997 (NLSY97) data are augmented with alcohol advertising, originating on the market level, for five media. The large sample of the MTF allows estimation of race and gender-specific models. The longitudinal nature of the NLSY97 allows controls for unobserved heterogeneity with state-level and individual fixed effects. Price and advertising effects are generally larger for females relative to males. Controls for individual heterogeneity yield larger advertising effects, implying that the MTF results may understate the effects of alcohol advertising. Results from the NLSY97 suggest that a 28% reduction in alcohol advertising would reduce adolescent monthly alcohol participation from 25% to between 24 and 21%. For binge participation, the reduction would be from 12% to between 11 and 8%. The past month price-participation elasticity is estimated at -0.26, consistent with prior studies. The results show that reduction of alcohol advertising can produce a modest decline in adolescent alcohol consumption, though effects may vary by race and gender. PMID- 16475247 TI - Swedish roadtrip. PMID- 16475248 TI - Doing business with the NIH. PMID- 16475249 TI - Current awareness in prenatal diagnosis. PMID- 16475250 TI - How to measure national stereotypes? PMID- 16475252 TI - Information technology in practice: lifestyle management and research. PMID- 16475251 TI - Achalasia of the cardia: dilatation or division? Is pneumatic balloon dilatation justifiable anymore? PMID- 16475253 TI - Legal and social aspects of biotechnology in Poland. PMID- 16475254 TI - Plasma lipid, lipoprotein and apolipoprotein levels in a random population sample of 2875 Hong Kong Chinese adults and their implications (NCEP ATP-III, 2001 guidelines) on cardiovascular risk assessment. AB - The age- and sex-related levels of plasma lipids, lipoproteins and apolipoproteins in a random population sample of 2875 Hong Kong Chinese Adults (1397 men and 1478 women aged 25-74) and their implications on cardiovascular risk assessment are reported. Total cholesterol, low-density lipoprotein (LDL) cholesterol and triglycerides increased with age in both sexes. Postmenopausal women had the worst profiles. They also showed higher triglyceride and non-high density lipoprotein (non-HDL)-cholesterol and had higher percentage of values greater than the desirable limits, compared with men of the same age groups. Overall 39% of men and 29% of women had non-HDL cholesterol of 4.2 mmol/L or greater. Apolipoproteins A-I and B followed the same trends as HDL-cholesterol and LDL-cholesterol, respectively. Apolipoprotein E (apo E) allele frequencies were: epsilon2 8.7, epsilon3 80.4 and epsilon4 10.9% with the genotype having a significant effect on plasma apo E concentration (p < 0.001). Carriers of the epsilon2 allele had higher apo E values than those homozygous for E3. Lipoprotein(a) levels were higher in women than men (geometric mean 152 versus 102 mg/L, p < 0.05) and in women with FSH above versus below 40 IU/L (185 versus 136 mg/L, p < 0.05). With respect to the NCEP ATP-III 2001 guidelines, the prevalence of hyperlipidemia in the Hong Kong population approached those in high CHD prevalence Caucasian communities. Local management guidelines and community wide programs to reduce fat intake, increase regular moderate exercise and reduce the prevalence of overweight and obesity are urgently required, and hormone replacement therapy for postmenopausal women might be warranted. PMID- 16475255 TI - [New primary immune deficiencies associated with combined variable immune deficiencies]. PMID- 16475256 TI - [Infectious agents and immune deficiencies]. PMID- 16475258 TI - [Kawasaki disease: an infectious disease?]. PMID- 16475257 TI - [Diagnosis of primary immune deficiencies in adults]. PMID- 16475259 TI - [Systemic manifestations and autoimmune diseases in primary immune deficiencies]. PMID- 16475260 TI - [Immunoglobulins and immune deficiency: methods of administration]. PMID- 16475261 TI - [Safety of medications derived from blood vis-a-vis new infectious agents]. PMID- 16475262 TI - [Infections and autoimmunity]. PMID- 16475264 TI - [Comprehension and prevention of infections using TNF inhibitors]. PMID- 16475263 TI - [Infections and lymphoma during prolonged immunosuppression in vascular diseases]. PMID- 16475265 TI - [Immunologic effects of anti-B-lymphocyte antibodies]. PMID- 16475266 TI - The tenth annual HIV drug guide. PMID- 16475267 TI - A guide to using this guide. PMID- 16475268 TI - Misinformation in HIV/AIDS care. There's more than one way to skin a virus. PMID- 16475269 TI - [Role of arsenolite on 8-isoprostane of asthmatic mice plasm]. AB - OBJECTIVE: Through the establishment of mouse' ovalbumin- sensitized asthmatic model and the observation of the 8-Isoprostane of plasm, to evaluate the therapeutic effects of arsenolite on asthmatic mice. METHOD: Forty-two healthy Kunming male mice were randomly divided into control group and experience groups, the latter were treated with dexamethasone, arsenolite. Lung function were tested, 8-isoprostane of plasm and WBC of BALF were measured. RESULT: Lung function improved after treating with dexamethasone or arsenolite. The WBC of asthmatic mice were significantly higher than those in control group, and decreased after treating with dexamethasone or arsenolite; 8-Isoprostane of plasm in asthmatic mice was higher than that of control group, and decreased after treating with dexamethasone or arsenolite. CONCLUSION: There is oxidant stress status in asthmatic mice. Arsenolite could lighten airway obstruction, reduce airway high response and redress oxidant stress status in asthmatic mice. PMID- 16475270 TI - [Alternations of ADF and G-actin in renal tubular epithelial cell of neonatal rats during kidney ischemic Injury]. AB - OBJECTIVE: To study the changes of ADF and G-actin in renal tubular epithelial cell of neonatal rats during kindney ischemic injury. To explore the interactions between ADF and G-actin and their relationships with actin cytoskeleton. METHODS: Twenty-four neonatal rats were used to the experiments due to have the weights from 30 to 40 g respectively. Renal ischemia was induced by clamping the left renal pedicle for different periods of time that was in 10 min or 30 min. The kidneys of normal neonatal rats were functioned as the control group. Two kinds of primary antibodies, which one was rabbit anti-chick ADF antiserum and another was mouse monoclonal G-actin antibody, were used on one same section of kidney tissue. FITC-labeled and TRITC-labeled secondary antibodies were used to identify ADF and G-actin respectively. These samples were examined by using a fluorescene microscope. RESULTS: In control group, ADF and G-actin showed to be diffused in cytoplasm of renal tubular epithelial cell. In ischemic injury group, the distributions of ADF and G-actin were changed significantly to apical region of tubular epithelial cells and the renal tubula lumen. Regardless of the ADF expression to not change but G-actin was increased after kidney ischemia. CONCLUSION: Under the physiolgical condition, ADF and G-actin got a diffuse distribution in cytoplasm of renal tubular epitbelial cell. However, when the kidney got in ischemia, the distributions of ADF and G-actin were changed significantly to apica region of tubular epithelial cell or the renal tubular lumen. PMID- 16475271 TI - [Effects of N-linked oligosaccharide chains on expression of integrin beta1 and apoptosis in glomerular mesangial cells]. AB - OBJECTIVE: The effects of N-linked oligosaccharide chains (TM), an inhibitor of N glycosylation of proteins, on the expression of beta1 integrin and the apoptosis of glomerular mesangial cells (GMC) were observed for exploring the role of inhibitors of N-linked oligosaccharide chains in cell apoptosis, proliferation, and expression of beta1 integrin and cyclin D1, and hence finding a new approach to the synteresis of proliferative nephropathy. METHODS: Cultured rat mesangial cells were divided into 4 groups: control group, and 0.5, 1.0 and 2.0 microg/mL TM-treatment groups. The expression of beta1 integrin and apoptosis rate were measured by flow cytometry; the expression of cyclin D, was measured by immunohistochemistry, and proliferation of GMCs was measured by MTT. RESULTS: The expression of beta1 integrin and cyclin D1 were decreased notably by inhibitors of N-linked oligosaccharide chains-TM. TM increased apoptosis and decreased the proliferative abilitiy of cells, and all the effects of TM were dose-dependent. CONCLUSION: Through repressed glycosylation of glycoprotein, TM can suppress the expression of beta1, integrin, affect the adhesion of cells, and increase the apoptosis rate; at the same time, the expression of cyclin D1 and the proliferative ability of cells were decreased, and all these were dose-dependent. PMID- 16475272 TI - Urease-induced alkalinization of extracellular pH and its antitumor activity in human breast and lung cancers. AB - Jack bean urease catalyzes the decomposition of urea into ammonia, which in turn increases the pH of the surrounding medium. Based on these two properties, we have investigated the antitumor effects of urease in vitro and in vivo on human lung and breast cancer cell lines either by the enzyme itself or in combination with other chemotherapeutic drugs. First, through the generation of toxic ammonia, urease exerted direct cytotoxicity on A549 and MDA-MB-231 tumor cells with LC50 of 0.22 and 0.45 U/ml, respectively. The cytotoxic effects could effectively be blocked using the reversible urease inhibitor acetohydroxamic acid. Complete protection was observed at dose > or = 2 mM. In addition, nude mouse xenograft models demonstrated that intratumoral urease injections (1 - 10 U/dose) inhibited A549 and MCF-7 tumor growth in vivo. Second, when combined with weak-base anticancer drugs, urease provided indirect antitumor effects via pH augmentation. Alkalinization of extracellular pH by urease (2 U/ml) and urea (> or = 2 mM) was found to enhance the antitumor efficacy of doxorubicin (50 microM) and vinblastine (100 microM) significantly. PMID- 16475273 TI - The failure of formal rights and equality in the clinic: a critique of bioethics. AB - For communities which espouse egalitarian principles, the hierarchical nature of care-giving relationships poses an extraordinary challenge. Patients' accounts of their illnesses and of their medical care capture the latent tension which exists between notional, political equality and the need for dependency on care from others. I believe that the power imbalance in doctor-patient relationships has broad implications for liberal democracies. Professional and care-giving relationships almost always consist of an imbalance of knowledge and expertise which no template of egalitarian moralism can suppress. When we seek help or guidance from authority figures, we are at a disadvantage politically even though we may be equal citizens theoretically and legally. Hierarchic relationships persist within democracies. Moreover, they tend to exist within a realm of privacy which is only partially visible from the social realm. In the end, traditional notions of liberal autonomy and egalitarianism do not properly describe or monitor these interactions. Liberal rhetoric (i.e., terms such as equality, rights, consent, etc.) pervades much of bioethical literature and interventions but, this very language tends to mask the persistence of structural hierarchies in the clinic. The doctor-patient relationship forces democratic communities to confront the problem of continuing hierarchic power relations and challenges liberalism to revise its understanding of individual autonomies. PMID- 16475274 TI - Clinical ethics case consultation. PMID- 16475275 TI - Death of John Paul II and the basic human care for the sick and the dying. AB - The death of Terri Schiavo by starvation and its sanction by some United States Courts indicates the alarming revival of the eugenics and euthanasia movement. From the legal sanction of physician-assisted suicide, the euthanasia movement now tries to advance the legal protection for "mercy killing." Terri was diagnosed with persistent vegetative state, a term that is outdated, vague and imprecise and that likens a human being to a vegetable. Medical literature indicates that patient with so-called "persistent vegetative state" can recover, and that they do experience pain. The euthanasia movement, linked to eugenics in its origin and present day influence in bioethics espouses the Nazi notion of "lives not worth living," unlimited patient autonomy, and philosophical utilitarianism. John Paul II countered the eugenic philosophy with the classical Western concept of man as the image and likeness of God, responsible for the care of himself and society as a whole. He taught in writing and by example that food and water are basic human care that every person should receive. In the last days of his life he showed a judicious use of proportionate or ordinary means to maintain life. He chose to forego disproportionate medical treatment when there was no reasonable hope of recovery. At that point he continued to receive ordinary medical care, together with basic human and spiritual care. PMID- 16475276 TI - COST B23 Conference (European Cooperation in the Field of Scientific and Technical Research), Debrecen, Hungary, October 2004. Abstracts. PMID- 16475277 TI - Clinical evaluation of impingement: what to do and what works. PMID- 16475278 TI - Biotechnology update: news and views. PMID- 16475279 TI - UNESCO bioethics declaration. PMID- 16475280 TI - Thermostable xylanase from Marasmius sp.: purification and characterization. AB - We have screened 766 strains of fungi from the BIOTEC Culture Collection (BCC) for xylanases working in extreme pH and/or high temperature conditions, the so called extreme xylanases. From a total number of 32 strains producing extreme xylanases, the strain BCC7928, identified by using the internal transcribed spacer (ITS) sequence of rRNA to be a Marasmius sp., was chosen for further characterization because of its high xylanolytic activity at temperature as high as 90 degrees C. The crude enzyme possessed high thermostability and pH stability. Purification of this xylanase was carried out using an anion exchanger followed by hydrophobic interaction chromatography, yielding the enzyme with >90% homogeneity. The molecular mass of the enzyme was approximately 40 kDa. The purified enzyme retained broad working pH range of 4-8 and optimal temperature of 90 degrees C. When using xylan from birchwood as substrate, it exhibits Km and Vmax values of 2.6 +/- 0.6 mg/ml and 428 +/- 26 U/mg, respectively. The enzyme rapidly hydrolysed xylans from birchwood, beechwood, and exhibited lower activity on xylan from wheatbran, or celluloses from carboxymethylcellulose and Avicel. The purified enzyme was highly stable at temperature ranges from 50 to 70 degrees C. It retained 84% of its maximal activity after incubation in standard buffer containing 1% xylan substrate at 70 degrees C for 3 h. This thermostable xylanase should therefore be useful for several industrial applications, such as agricultural, food and biofuel. PMID- 16475281 TI - Evidence that autobiographic memory retrieval does not become independent of the hippocampus: an fMRI study contrasting very recent with remote events. AB - Traditional consolidation theory, which seeks to explain how new memories are incorporated into the preexisting neural architecture, stipulates that the hippocampus plays a time-limited role in this process. However, although there is abundant research showing that the hippocampus is necessary for the initial (encoding) phase, there is very little experimental evidence with human subjects proving that the structure ceases to play a role in the retrieval of episodic items from memory stores. To test this hypothesis, we investigated recall activation associated with recent memories (2.5 days) versus remote memories (mean of 8 years) using functional magnetic resonance imaging. In accordance with the multiple memory trace theory, recall of consolidated autobiographic information, represented by the remote condition, was not independent of the hippocampus. Both types of memory retrieval produced significant activation in parahippocampal, prefrontal, and midtemporal gyri, the parietal-temporal junction, and a medial region of cortex spanning the posterior cingulate and precuneus gyri. However, where recent events activated bilateral regions of the caudate nucleus, remote events yielded significantly greater activation within the hippocampus proper. The results challenge traditional consolidation theory, which would predict greater hippocampal activity for recent events. Furthermore, they highlight the interplay between multiple memory systems in the brain. We argue that our particular question format, which encouraged depth of recall and did not require a prescan interview, as well as our delineation of the recent and remote time periods, were the determining factors for the observed pattern of hippocampal activation. PMID- 16475282 TI - How intravenous supplies, will travel. PMID- 16475283 TI - Time in the reproduction of mortuary practices. PMID- 16475284 TI - Zooarchaeology: past, present and future. PMID- 16475285 TI - A critical overview of archaeological animal bone studies. PMID- 16475286 TI - Intimate sexual relations in prehistory: lessons from the Japanese macaques. PMID- 16475287 TI - Demographic growth, environmental changes and technical adaptations: responses of an agricultural community from the 32nd to the 30th centuries BC. PMID- 16475288 TI - Tiwanaku 'colonization': bioarchaeological implications for migration in the Moquegua Valley, Peru. PMID- 16475289 TI - Using stable nitrogen-isotopes to study weaning behavior in past populations. PMID- 16475291 TI - The world on a plate: ceramics, food technology and cosmology in Neolithic Orkney. PMID- 16475290 TI - Food technologies/technologies of the body: the social context of wine and oil production and consumption in Bronze Age Crete. PMID- 16475292 TI - The social context of food technology in Iron Age Gao, Mali. PMID- 16475293 TI - Archaeological biographies: realizing human lifecycles, -courses and -histories. PMID- 16475294 TI - The evolution of human life history. PMID- 16475295 TI - Neanderthal lifecycles: developmental and social phases in the lives of the last archaics. PMID- 16475296 TI - Cycles of life and death: narrative homology and archaeological realities. PMID- 16475297 TI - Girling the girl and boying the boy: the production of adulthood in ancient Mesoamerica. PMID- 16475298 TI - Agriculture and dental caries? The case of rice in prehistoric Southeast Asia. PMID- 16475299 TI - 'Doing trade': a sexual economy of nineteenth-century Australian female convict prisons. PMID- 16475301 TI - Stable carbon and nitrogen isotopic analyses of the underclass at the colonial Cape of Good Hope in the eighteenth and nineteenth centuries. PMID- 16475302 TI - Decorous bodies and disordered passions: representations of emotion among the Classic Maya. PMID- 16475304 TI - Archaeology and evolutionary ecology. PMID- 16475305 TI - Subsistence strategies and early human population history: an evolutionary ecological perspective. PMID- 16475306 TI - Evolutionary ecology of spoken language: co-evolutionary hypotheses are testable. PMID- 16475307 TI - Evaluating evolutionary archaeology. PMID- 16475308 TI - Thinking outside the box: a new perspective on diet breadth and sexual division of labor in the Prearchaic Great Basin. PMID- 16475309 TI - [60 years of the journal "Medicinski arhiv"]. PMID- 16475310 TI - Bacteria contribute to freshwater PCB contamination. PMID- 16475311 TI - Regulating the underground injection of CO2. PMID- 16475312 TI - Transgenic plants in phytoremediation: recent advances and new possibilities. AB - Phytoremediation, the use of plants and their associated microbes to remedy contaminated soils, sediments, and groundwater, is emerging as a cost-effective and environmentally friendly technology. Due in large part to its aesthetic appeal, this technology has gained increasing attention over the past 10 years. Phytoremediation uses different plant processes and mechanisms normally involved in the accumulation, complexation, volatilization, and degradation of organic and inorganic pollutants. Certain plants, called hyperaccumulators, are good candidates in phytoremediation, particularly for the removal of heavy metals. Phytoremediation efficiency of plants can be substantially improved using genetic engineering technologies. Recent research results, including overexpression of genes whose protein products are involved in metal uptake, transport, and sequestration, or act as enzymes involved in the degradation of hazardous organics, have opened up new possibilities in phytoremediation. This paper provides a critical review of the recent progress made toward the development of transgenic plants with improved phytoremediation capabilities and their potential use in environmental cleanup. PMID- 16475313 TI - Perchlorate and nitrate in leafy vegetables of North America. AB - In previous studies trace levels of perchlorate were found in lettuce (Lactuca sativa L.) irrigated with Colorado River water, which is contaminated with low levels of perchlorate from aerospace and defense related industries. In this paper, we report the results of a survey conducted across North America to evaluate the occurrence of perchlorate in leafy vegetables produced outside the lower Colorado River region, and evaluate the relative iodide uptake inhibition potential to perchlorate and nitrate in these leafy vegetables. Conventionally and organically produced lettuce and other leafy vegetable samples were collected from production fields and farmers' markets in the central and coastal valleys of California, New Mexico, Colorado, Michigan, Ohio, New York, Quebec, and New Jersey. Results show that 16% of the conventionally produced samples and 32% of the organically produced samples had quantifiable levels of perchlorate using ion chromatography. Estimated perchlorate exposure from organically produced leafy vegetables was approximately 2 times that of conventional produce, but generally less than 10% of the reference dose recommended by the National Academy of Sciences. Furthermore, the iodide uptake inhibition potential of perchlorate was less than 1% of that of the nitrate present. These data are consistent with those of other reported perchlorate survey work with lettuce, bottled water, breast milk, dairy milk, and human urine, and suggest a wide national presence of perchlorate. PMID- 16475314 TI - XAS evidence of As(V) association with iron oxyhydroxides in a contaminated soil at a former arsenical pesticide processing plant. AB - The molecular-level speciation of arsenic has been determined in a soil profile in the Massif Central near Auzon, France that was impacted by As-based pesticides by combining conventional techniques (XRD, selective chemical extractions) with X ray absorption spectroscopy (XAS). The arsenic concentration is very high at the top (>7000 mg kg(-1)) and decreases rapidly downward to a few hundreds of milligrams per kilogram. A thin layer of schultenite (PbHAsO4), a lead arsenate commonly used as an insecticide until the middle of the 20th century, was found at 10 cm depth. Despite the occurrence of this As-bearing mineral, oxalate extraction indicated that more than 65% of the arsenic was released upon dissolution of amorphous iron oxides, suggesting a major association of arsenic with these phases within the soil profile. Since oxalate extraction cannot unambiguously distinguish among the various chemical forms of arsenic, these results were confirmed by a direct in situ determination of arsenic speciation using XAS analysis. XANES data indicate that arsenic occurs mainly as As(V) along the soil profile except for the topsoil sample where a minor amount (7%) of As(III) was detected. EXAFS spectra of soil samples were fit by linear combinations of model compounds spectra and by a shell-by-shell method. These procedures clearly confirmed that As(V) is mainly (at least 80 wt %) associated with amorphous Fe(III) oxides as coprecipitates within the soil profile. If any, the proportion of schultenite, which was evidenced by XRD in a separate thin white layer, does not account for more than 10 wt % of arsenic in soil samples. This study emphasizes the importance of iron oxides in restricting arsenic dispersal within soils following dissolution of primary As-bearing solids manufactured for use as pesticides and released into the soils. PMID- 16475315 TI - Atmospheric PCB concentrations at Terra Nova Bay, Antarctica. AB - Concentrations of gas-phase polychlorobiphenyls (PCBs) were studied over an austral summer at a site in Terra Nova Bay, Antarctica. Gas-phase concentrations of individual PCB congeners in the atmosphere of Terra Nova Bay ranged from below the detection limit to 0.25 pg m(-3), with a mean concentration of sigmaPCB of 1.06 pg m(-3). The PCB profile was dominated by lower-chlorinated PCB congeners; in fact >78% of the total PCB content was due to congeners with 1-4 chlorine atoms and only about 10% with 5-7 chlorines, whereas higher-chlorinated PCB congeners were below detection limits. The mean sigmaPCB concentration obtained in this study were lower than those reported in previous Antarctic studies. Temporal concentration profiles of sigmaPCB do not correspond to seasonal temperature changes. In consideration of the low PCB concentrations observed, the studies with the wind roses, the regression between In P(PCB) and T(-1), and the distribution of congeners, we can hypothesize that PCB local source contributions are not very important, whereas long-distance transport is the prevalent factor bringing PCBs to Terra Nova Bay. PMID- 16475316 TI - Development of an ecosystem sensitivity model regarding mercury levels in fish using a preference modeling methodology: application to the Canadian boreal system. AB - The objective of this study is to use a preference modeling methodology as a predictive tool to roughly assess the sensitivity of the ecosystems regarding mercury (Hg) concentrations in fish. We apply a preference modeling methodology to rank lakes within the boreal forest from highest to lowest Hg concentrations in fish using simple environmental factors. Among the numerous variables influencing Hg fate in the environment, we only retain simple key indicators that are expected to influence Hg concentrations in fish tissue such as watershed characteristics of the lake (percentage of the catchment area of the lake, ratio of drainage area versus lake area, percentage of the drainage area of the lake as wetlands, land use, and clear-cutting), lake characteristics (chlorophyll, dissolved organic carbon, pH, and fishing intensity), and atmospheric Hg inputs. Preliminary results of modeling that we carried out using a set of Canadian lakes of boreal forest data are promising. With only a minimum set of criteria, we are able to reproduce the trends of Hg contamination in fish caught in six regions of the Canadian boreal forest and classify the sensitivity of the ecosystems to Hg loadings in three categories: high, medium, and low. PMID- 16475317 TI - Cold temperature PM emissions measurement: method evaluation and application to light duty vehicles. AB - This work examines the methodology to sample and measure the number and size of motor vehicle particulate emissions (PM) at subambient temperatures. The study has two principal objectives. The first is to address the following question: which aspects of the particle sampling, dilution, and size measurement process must be made at the vehicle test temperature to obtain an accurate representation of the PM emissions? The second is to perform a preliminary overview of how subambient temperature operation affects PM emissions from the major classes of current model light duty vehicles. The principal findings are the following: (1) The temperature of the particle size instruments, test cell versus room temperature, has little effect on the measurements. (2) Once the engine has warmed, solid particle (soot) mode emissions in the cold test cell are similar to those at room temperature. The first finding simplifies cold temperature emissionstesting because it allows particle sizing instruments to be placed outside the cold test cell and operated at room temperature. The latter is consistent with the expectation that solid particles are formed in the engine and are therefore relatively unaffected by ambient conditions after engine warm-up. Use of cold dilution air in the room-temperature test cell increases the number and size of nuclei particles; however, the effect of dilution airtemperature was inconclusive in the cold test cell. PMID- 16475318 TI - Geographical, spatial, and temporal distributions of multiple indoor air pollutants in four Chinese provinces. AB - Exposure to indoor air pollution from household energy use depends on fuel, stove, housing characteristics, and stove use behavior. We monitored three important indoor air pollutants-respirable particles (RPM), carbon monoxide (CO), and sulfur dioxide (SO2)-for a total of 457 household-days in four poor provinces in China (Gansu, 129 household-days; Guizhou, 127 household-days; Inner Mongolia, 65 household-days; and Shaanxi, 136 household-days), in two time intervals during the heating season to investigate spatial and temporal patterns of pollution. The two provinces where biomass is the primary fuel (Inner Mongolia and Gansu) had the highest RPM concentrations (719 microg/m3 in the single cooking/living/bedroom in Inner Mongolia in December and 351-661 microg/m3 in different rooms and months in Gansu); lower RPM concentration were observed in the primarily coal-burning provinces of Guizhou and Shaanxi (202-352 microg/m3 and 187-361 microg/m3 in different rooms and months in Guizhou and Shaanxi, respectively). Inner Mongolia and Gansu also had higher CO concentrations (7.4 ppm in the single cooking/living/bedroom in Inner Mongolia in December and 4.8 11.3 ppm in different rooms and months in Gansu). Among the two primarily coal burning provinces, Guizhou had lower concentrations of CO than Shaanxi (1.2-1.8 ppm in Guizhou vs 2.0-13.3 ppm in different rooms and months in Shaanxi). In the two coal-burning provinces, SO2 concentrations were substantially higher in Shaanxi than in Guizhou. Relative concentrations in different rooms and provinces indicate that in the northern provinces heating is an important source of exposure to indoor pollutants from energy use. Day-to-day variability of concentrations within individual households, although substantial, was smaller than variation across households. The implications of the findings for designing environmental health interventions in each province are discussed. PMID- 16475319 TI - Spatially complex distribution of dissolved manganese in a fjord as revealed by high-resolution in situ sensing using the autonomous underwater vehicle Autosub. AB - Loch Etive is a fjordic system on the west coast of Scotland. The deep waters of the upper basin are periodically isolated, and during these periods oxygen is lost through benthic respiration and concentrations of dissolved manganese increase. In April 2000 the autonomous underwater vehicle (AUV) Autosub was fitted with an in situ dissolved manganese analyzer and was used to study the spatial variability of this element together with oxygen, salinity, and temperature throughout the basin. Six along-loch transects were completed at either constant height above the seafloor or at constant depth below the surface. The ca. 4000 in situ 10-s-average dissolved Mn (Mnd) data points obtained provide a new quasi-synoptic and highly detailed view of the distribution of manganese in this fjordic environment not possible using conventional (water bottle) sampling. There is substantial variability in concentrations (<25 to >600 nM) and distributions of Mnd. Surface waters are characteristically low in Mnd reflecting mixing of riverine and marine end-member waters, both of which are low in Mnd. The deeper waters are enriched in Mnd, and as the water column always contains some oxygen, this must reflect primarily benthic inputs of reduced dissolved Mn. However, this enrichment of Mnd is spatially very variable, presumably as a result of variability in release of Mn coupled with mixing of water in the loch and removal processes. This work demonstrates how AUVs coupled with chemical sensors can reveal substantial small-scale variability of distributions of chemical species in coastal environments that would not be resolved by conventional sampling approaches. Such information is essential if we are to improve our understanding of the nature and significance of the underlying processes leading to this variability. PMID- 16475320 TI - Identification of brominated carbazoles in sediment cores from Lake Michigan. AB - During the analysis of brominated flame retardants in sediment cores taken from Lake Michigan, a group of unknown brominated compounds were observed. To identify their chemical structures, a variety of gas chromatographic mass spectrometric techniques were used. With the help of high-resolution and positive chemical ionization mass spectrometry, the molecular formula of the most abundant congener was established to be C12H5Br4N. On the basis of the electron impact and electron capture negative ionization mass spectra, this chemical was suspected of being a tetrabromocarbazole. The structure was confirmed by comparing the mass spectra and gas chromatographic retention times of an authentic standard of 1,3,6,8 tetrabromocarbazole with those of the most abundant unknown. The other unknown compounds may be other bromocarbazole congeners. The time trend determined by measuring these compounds as a function of depth in the Lake Michigan sediment cores indicated that the concentration of 1,3,6,8-tetrabromocarbazole peaked around 1920-1935. This is the first time that bromocarbazoles have been found in the environment. PMID- 16475321 TI - Role of chlorine in combustion field in formation of polychlorinated dibenzo-p dioxins and dibenzofurans during waste incineration. AB - Combustion experiments performed in the presence of hydrogen chloride (HCl) in a laboratory-scale fluidized-bed reactor were carried out to elucidate the role of chlorine in the formation of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs; together: PCDD/Fs) in various sections of a municipal waste incinerator. We first demonstrated that the homologue profile and the pattern of the congener proportions of PCDD/Fs for a model waste containing poly(vinyl chloride) (PVC) combusted in the absence of HCl were similar to those for a PVC free waste combusted in the presence of HCl. This showed no difference between PVC in the waste and injected HCl in the role as a chlorine source in PCDD/F formation during incineration. Next, to investigate PCDD/F formation in each section of the incinerator, we carried out combustion experiments with the PVC free waste, injecting HCl at different locations of the incinerator. The amounts of PCDDs and PCDFs formed were significantly reduced when HCI was not supplied to the main combustion section. The presence of HCI in the main combustion section was essential for the formation of PCDD/Fs, even in the downstream sections. This finding indicates that compounds that were able to form PCDD/Fs in the downstream sections were mainly formed in the main combustion section in the presence of HCl. PMID- 16475322 TI - Concentrations of polybrominated diphenyl ethers, polychlonnated biphenyls, and polychlorobiphenylols in serum from pregnant Faroese women and their children 7 years later. AB - The objective of this study was to assess blood concentrations of polybrominated diphenyl ethers (PBDEs) and polychlorinated biphenyls (PCBs), and their polychlorobiphenylol (OH-PCB) metabolites in humans with a high seafood intake. Samples were obtained from pregnant women in the Faroe Islands in 1994-1995 and from their children at 7 years of age to examine maternal transfer of the compounds to their child, age-dependent metabolism, and temporal changes. Maternal serum was dominated by 2,2',4,4'-tetrabromodiphenyl ether (BDE-47), while 2,2',4,4',5,5'-hexabromodiphenyl ether (BDE-153) prevailed in the children's serum seven years later. DecaBDE was present in both mothers and children up to 3 and 6 ng/g lipid weight, respectively. The sigmaPCB concentration in the children averaged about 60% of the concentrations in their mothers, with median levels for both above 1 microg/g lipid weight and .with similar PCB congener patterns. sigmaOH-PCB serum concentrations from the mothers and their children showed ranges of 1.8-36 ng/g wet weight (ww) and 0.49-22 ng/g ww, respectively, with all OH-PCB congener concentrations being lower in the children, except for 2,3,3',4',5-pentachloro-4-biphenylol (4-OH-CB107). Children at 7 years of age are exposed to PCBs at levels only slightly below those of their mothers, and the increased 4-OH-CB107 concentrations in children could be due to age-related differences in PCB metabolism. The PBDE concentrations were similar in both mothers and their children. The main persistent organic pollutant concentrations in the children are most probably due to other environmental exposure than maternal transfer. PMID- 16475323 TI - Platinum group element concentrations and osmium isotopic composition in urban airborne particles from Boston, Massachusetts. AB - Platinum, Pd, Rh, and Os were found to occur at elevated concentrations in airborne particles (PM10) collected at urban sites in Boston, MA. Average Pt, Pd, Rh, and Os concentrations were 6.9 +/- 1.9, 8.1 +/- 1.8, 1.50 +/- 0.50, and 0.068 (-0.068 + 0.070) pg m(-3), respectively. Elevated Pt, Pd, and Rh concentrations are attributed to automobile catalysts, which use Pt, Pd, and Rh for the removal of pollutants from engine exhaust gas. An automobile catalyst source is supported by significant correlations between these elements and by a Pt/Rh similar to that in catalysts. Elevated Os concentrations are also believed to be the result of emission from automobile catalysts in which Os occurs as an impurity. The isotopic composition of Os (187Os/ 188Os) ranged from 0.30 to 2.90, indicating large variations in Os sources. Osmium has a predominantly anthropogenic origin at concentrations > 0.1 pg m(-3), whereas natural sources are more important at lower Os concentrations. Osmium isotopic composition also indicates that Pt, Pd, and Rh in Boston air are of almost exclusive anthropogenic origin, with a relatively small natural contribution. Our results indicate that scavenging by rain plays a major role in the atmospheric residence time and environmental fate of PGE. PMID- 16475324 TI - Photodegradation of dimethyl sulfide (DMS) in natural waters: laboratory assessment of the nitrate-photolysis-induced DMS oxidation. AB - The interaction of sunlight and dissolved chromophoric matter produces reactive chemical species that are significant in the removal of dimethyl sulfide (DMS) in the surface ocean. Using artificial solar radiation, we examined the role of several inorganic components of seawater on the kinetics of NO3- -photolysis induced DMS removal in aqueous solution. This study strongly suggests that NO3- photolysis products react significantly with DMS in aqueous solution possibly via an electrophilic attack on the electron-rich sulfur atom. This supports previous field observations that indicate that NO3- photolysis has a substantial control on DMS photochemistry in nutrient-rich waters. A key finding of this research is that the oxidation rate of DMS induced by NO3- photolysis is dramatically enhanced in the presence of bromide ion. Moreover, our results suggest that bicarbonate/carbonate ions are involved in free radical production/scavenging processes important for DMS photochemistry. These reactions are pH dependent. We propose that DMS removal by some selective free radicals derived from bromide and bicarbonate/carbonate ion oxidation is a potentially important and previously unrecognized pathway for DMS photodegradation in marine waters. PMID- 16475325 TI - XAS study of iron and arsenic speciation during Fe(II) oxidation in the presence of As(III). AB - The speciation of As and Fe was studied during the oxidation of Fe(II)-As(III) solutions by combining XAS analysis at both the Fe and As K-edges. Fe(II) and As(III) were first hydrolyzed to pH 7 under anoxic conditions; the precipitate was then allowed to oxidize in ambient air for 33 h under vigorous stirring. EXAFS analysis at the As K-edge shows clear evidence of formation of inner-sphere complexes between As(III) and Fe(II), i.e., before any oxidation. Inner-sphere complexes were also observed when Fe became sufficiently oxidized, in the form of edge-sharing and double-corner linkages between AsIIIO3 pyramids and FeIIIO6 octahedra. XAS analyses at the Fe K-edge reveal that the presence of As(III) in the solution limits the polymerization of Fe(II) and the formation of green rust and inhibits the formation of goethite and lepidocrocite. Indeed, As(III) accelerates the Fe(II) oxidation kinetics and leads to the formation of nanosized Fe-As subunits of amorphous aggregates. These observations, rather than a presumed weaker affinity of As(III) for iron oxyhydroxides, might explain why As(III) is more difficult to remove than As(V) by aerating reducing groundwater. PMID- 16475326 TI - Enhanced coagulation due to evaporation and its effect on nanoparticle evolution. AB - A new explanation for the evolution of particles near a roadway is proposed. The explanation starts with data that suggest that small (<15 nm) liquid nanoparticles shed semivolatile organics ( or =10(8)-10(9) and 10(7) 10(8) mol(-1) L, respectively. In effluent from sulfate-reducing bioreactors, Co is also found to be present in a strongly bound form, and up to micromolar levels of strongly complexing excess ligand was detected. The predominant presence of Co and Ni in strong complexes, with concomitant low free dissolved concentrations, is significant for limitation by these elements in anaerobic bioreactors. PMID- 16475328 TI - Effect of microbes on contaminant transfer in the Lake Superior food web. AB - The partitioning of PCBs to natural populations of aquatic heterotrophic bacteria from Lake Superior was measured in both field and laboratory studies to better understand the role of bacteria and the microbial food web in persistent, bioaccumulative, toxic organic contaminant (PBT) transfer. A filtration method that separated material >1 microm from that <1 microm was used to collect and measure PCB concentrations in the bacterial fraction. We used bacterial biovolume and a conversion factor to calculate bacterial carbon, which was used to normalize PCB concentrations. The range of total PCB concentrations in the bacterial fraction (61-337 ng/g organic carbon; OC) was similar, but generally higher than that of the total particulate fraction (36-324 ng/g OC). Measured log bioaccumulation factors (BAFoc, bacterial fraction; Koc, total particulate) were significantly greater in the bacterial fraction than in the total particulate fraction (bacterial range 6.7-7.3, total particulate range 5.8-7.0). Laboratory experiments demonstrated that a hexachlorobiphenyl reached steady state with a natural community of Lake Superior bacteria within 48 h and had a mean log BAFoc of 7.5 +/- 1.9. The octanol/ water partition coefficient (Kow) consistently under predicted BAFoc, however, the magnitude of the under-prediction was still within the range of uncertainty in food web modeling (factors of 3-9). Food web modeling and risk assessment of PBT bioaccumulation in aquatic systems could be improved by considering the microbial food web (bacteria and its protozoan grazers) as a previously unaccounted for pathway of contaminant transfer. PMID- 16475329 TI - Adsorption of sulfonamide antimicrobial agents to clay minerals. AB - Adsorption of three sulfonamide antimicrobials to clay minerals was investigated as a function of pH, ionic strength, and type of exchangeable cation. Sulfonamide antimicrobial adsorption exhibited pronounced pH dependence consistent with sorbate speciation and clay properties. Sulfonamide antimicrobials did not intercalate into montmorillonite, and surface charge density influenced sorption by determining adsorption domain size. Adsorption edge data were best fit to a model including terms for the cationic and uncharged species. Adsorption of uncharged sulfamethazine to montmorillonite was relatively insensitive to pH, ionic strength, and type of exchangeable cation, while that to kaolinite was highly sensitive to ionic strength. Adsorption of cationic sulfamethazine to montmorillonite exceeded that of the neutral species by 1-2 orders of magnitude, but was unimportant for kaolinite atthe pH values examined. Cation exchange appeared to contribute to sorption of cationic sulfonamide species to montmorillonite. Anionic sulfamethazine adsorption was negligible. The nature of the sulfonamide R group influenced the degree of adsorption of cationic and neutral species. Our results highlight the importance of considering sulfonamide speciation and clay surface charge density in predicting the transport of these antimicrobials. PMID- 16475330 TI - Microcosm experiments to assess the effects of temperature and microbial activity on polychlorinated biphenyl transport in anaerobic sediment. AB - Increased polychlorinated biphenyl (PCB) loadings from sediment to the water column under low-flow conditions during late spring and summer months have been observed in the Grasse River (Massena, NY) and other PCB-contaminated rivers in the United States. Temperature appears to be an important factor affecting this phenomenon, as increased sediment temperature leads to increased microbial and bioturbator activity as well as increased rates of diffusion and desorption. Laboratory-scale sediment microcosms were developed and used to study with fine scale resolution the effects of temperature and microbial activity on PCB transport in river sediment. Over the time course of these experiments, with the extraction procedures used, irreversible sorption of each congener to sediment was observed and increased with aging of the sediment. Temperature-dependent transport was observed for PCB congeners 2,4,5-trichlorobiphenyl (BZ29), 2,5 dichlorobiphenyl (BZ9), and 2-chlorobiphenyl (BZ1) in Grasse River sediment and a synthetic sediment system. The fastesttransport of the congeners occurred in biologically active Grasse River sediment followed by biologically inactive (autoclaved) Grasse River sediment, and synthetic sediment. The increased transport in biologically active sediment demonstrated the importance of microbial activity, in particular gas bubble generation, in PCB transport in near surface sediments. PMID- 16475331 TI - Adsorption of arsenate onto ferrihydrite from aqueous solution: influence of media (sulfate vs nitrate), added gypsum, and pH alteration. AB - Mineral processing effluents generated in hydrometallurgical industrial operations are sulfate based; hence it is of interest to investigate the effect sulfate matrix solution ("sulfate media") has on arsenate adsorption onto ferrihydrite. In this work, in particular, the influence of media (SO4(2-) vs NO3 ), added gypsum, and pH alteration on the adsorption of arsenate onto ferrihydrite has been studied. The ferrihydrite precipitated from sulfate solution incorporated a significant amount of sulfate ions and showed a much higher adsorption capacityfor arsenate compared to nitrateferrihydrite at pH 3-8 and initial Fe/As molar ratios of 2, 4, and 8. Adsorption of arsenate onto sulfate-ferrihydrite involved ligand exchange with SO4(2-) ions that were found to be more easily exchangeable with increasing pH. Added gypsum to the adsorption system significantly enhanced the uptake of arsenate by ferrihydrite at pH 8. Equilibration treatment at acidic pH and addition of gypsum markedly improved the stability of adsorbed arsenate on ferrihydrite when pH was elevated. Comparison of arsenate adsorption onto ferrihydrite to coprecipitation of arsenate with iron(III) showed the latter process to lead to higher arsenic removal. PMID- 16475332 TI - Fluorescence quenching and luminescence sensitization in complexes of Tb3+ and Eu3+ with humic substances. AB - Intrinsic fluorescence quenching of humic substances (HS) and the sensitization of Ln3+ luminescence (Ln3+ = Tb3+, Eu3+) in HS complexes were investigated. Both measurements yielded complementary information on the complexation of metals by HS. Large differences between fulvic acids (FA) and humic acids (HA) were found. From time-resolved luminescence measurements it is concluded that a combination of energy transfer and energy back transfer between HS and Ln3+ is responsible for the observed luminescence decay characteristics. In the case of Eu3+, an additional participation of charge-transfer states is suggested. A new concept for the evaluation of the sensitized luminescence decays of Ln3+ was adapted. PMID- 16475333 TI - Is glassiness a common characteristic of soil organic matter? AB - Until now, glass transitions were detected in isolated humic and fulvic acids as well as in distinct soil samples with usually high Corg contents. The results of this study indicate that glassiness has to be considered a common characteristic of soil organic matter (SOM). However, two types of glassiness were observed in various soil types. Additionally to a typical glass transition with low intensity, a slowly reversing glasstransition-like step transition with significantly higher intensity was detected in 52 out of 102 tested soil samples. The intensity of this transition type is correlated to the organic matter content of the samples. The transition behavior additionally depends on characteristics of the locations and changes within soil profiles. Relations to particulate organic matter (POM), mineral-associated organic matter (MOM), and the thermostable fraction of the soil samples were not significant. A surprising result of the study is that the step transition temperatures of all analyzed air dried soil samples range between 51 and 67 degrees C in closed systems, pointing to a superordinate mechanism which controls the matrix rigidity of the organic molecules. This may be represented by the formation of hydrogen bond based cross links between water molecules and SOM suggested in a previous study. Thus, glassiness in SOM may be caused by physical and physicochemical mechanisms. PMID- 16475334 TI - Photoirradiation of dissolved humic acid induces arsenic(III) oxidation. AB - The fate of arsenic in aquatic systems is influenced by dissolved natural organic matter (DOM). Using UV-A and visible light from a medium-pressure mercury lamp, the photosensitized oxidation of As(III) to As(V) in the presence of Suwannee River humic acid was investigated. Pseudo-first-order kinetics was observed. For 5 mg L(-1) of dissolved organic carbon (DOC) and 1.85 mEinstein m(-2) s(-1) UV-A fluence rate, the rate coefficient k degrees exp was 21.2 +/- 3.2 10(-5) s(-1), corresponding to a half-life <1 h. Rates increased linearly with DOC and they increased by a factor of 10 from pH 4 to 8. Based on experiments with radical scavengers, heavy water, and surrogates for DOM, excited triplet states and/or phenoxyl radicals seem to be important photooxidants in this system (rather than singlet oxygen, hydrogen peroxide, hydroxyl radicals, and superoxide). Photoirradiation of natural samples from freshwater lakes, rivers, and rice field water (Bangladesh) showed similar photoinduced oxidation rates based on DOC. Fe(III) (as polynuclear Fe(III)-(hydr)oxo complexes or Fe(III)-DOC complexes) accelerates the rate of photoinduced As(III) oxidation in the presence of DOC by a factor of 1.5-2. PMID- 16475335 TI - Human cell mutagens in respirable airborne particles from the northeastern United States. 2. Quantification of mutagens and other organic compounds. AB - Few reports have characterized mutagenic compounds in respirable airborne particles (<2.5 micrometers in diameter; PM2.5) collected at different sites on a regional scale (hundreds of km). Previously, we reported differences in the human (h1A1v2) cell mutagenicity of whole and fractionated organic extracts of PM2.5 samples collected in Boston, MA, Rochester, NY, and Quabbin Reservoir, a rural site in western MA. Herein we describe the analysis of mutagens and other organic compounds in these samples. Gas chromatography-mass spectrometry (GC-MS) was used to quantify approximately 150 organic compounds, including 31 known human cell mutagens. Molecular weight (MW) 226-302 amu PAHs were the most important mutagens identified: cyclopenta[cd]pyrene accounted for 1-2% of the measured mutagenicity of the samples, MW 252 PAHs accounted for 4-6%, MW 276-278 PAHs accounted for 2 5%, and MW 302 PAHs accounted for 2-3%. 6H-benzo-[cd]pyren-6-one, a PAH ketone, accounted for 3-5% of the mutagenicity. The same compounds accounted for similar portions of the total attributed mutagenicity in each sample. Mutagen levels were similar in the Boston and Rochester samples, and both were significantly higher than the Quabbin sample. This may explain whythe mutagenicities of the Boston and Rochester samples were higher than the Quabbin sample. The levels of mutagens found in semipolar fractions, however, could not explain why the mutagenicity of semipolar fractions was 2-fold higher in the Rochester sample than in the Boston sample. Known mutagens accounted for only 16-26% of the total mutagenicity of the unfractionated extracts, and only approximately 20% of the mutagenicity of the nonpolar and semipolar fractions. The remaining mutagenicity is likely attributable to other, as-yet unknown, semipolar and polar mutagens, or to interactions among chemical constituents of the samples. These findings are consistent with similar studies performed on airborne particles from Los Angeles and Washington, DC, thus indicating that PAHs, PAH-ketones, and as-yet unidentified polar organic compounds are widely distributed airborne human cell mutagens. PMID- 16475336 TI - Fluoride removal by calcite: evidence for fluorite precipitation and surface adsorption. AB - Fluoride contamination of groundwater, both anthropogenic and natural, is a major problem worldwide. In this study, fluoride removal by crushed limestone (99% pure calcite) was investigated by batch studies and surface-sensitive techniques from solutions with fluoride concentrations from 150 micromol/L (3 mg/L) to 110 mM (approximately 2100 mg/L). Surface-sensitive techniques, including atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS) as well as zeta potential measurements, confirm that, in addition to precipitation reactions, adsorption of fluoride also occurs. Results indicate that fluoride adsorption occurs immediately over the entire calcite surface with fluorite precipitating at step edges and kinks, where dissolved Ca2+ concentration is highest. The PHREEQ geochemical model was applied to the observed data and indicates that existing models, especially at low fluoride concentrations and high pH (>7.5) are not equipped to describe this complex system, largely because the PHREEQ model includes only precipitation reactions, whereas a combination of adsorption and precipitation parameters are required. PMID- 16475337 TI - Environmental microbes can speciate and cycle arsenic. AB - Naturally occurring arsenic is found predominantly as arsenate [As(V)] or arsenite [As(III)], and can be readily oxidized or reduced by microorganisms. Given the health risks associated with arsenic in groundwater and the interest in arsenic-active microorganisms, we hypothesized that environmental microorganisms could mediate a redox cycling of arsenic that is linked to their metabolism. This hypothesis was tested using an As(V) respiring reducer (strain Y5) and an aerobic chemoautotrophic As(II) oxidizer (strain OL1 ) both isolated from a Superfund site, Onondaga Lake, in Syracuse, NY. Strains were grown separately and together in sealed serum bottles, and the oxic/anoxic condition was the only parameter changed. Initially, under anoxic conditions when both isolates were grown together, 2 mM As(V) was stoichiometrically reduced to As(III) within 14 days. Following complete reduction, sterile ambient air was added and within 24 h As(III) was completely oxidized to As(V). The anoxic-oxic cycle was repeated, and sterile controls showed no abiotic transformation within the 28-day incubation period. These results demonstrate that microorganisms can cycle arsenic in response to dynamic environmental conditions, thereby affecting the speciation, and hence mobility and toxicity of arsenic in the environment. PMID- 16475339 TI - Kinetic mechanism for predicting secondary organic aerosol formation from the reaction of d-limonene with ozone. AB - A semi-explicit mechanism of d-limonene was developed and tested against experimental results obtained from large outdoor Teflon film chambers at the University of North Carolina (UNC) smog chamber facility. The model couples gas phase reactions with partitioning processes and possible particle-phase reactions. The model not only tracks the gas-phase ozonolysis reaction of d limonene, but also provides a reasonable prediction of the secondary aerosol mass production under different conditions. Limononaldehyde was the major identified product, followed by limona-ketone, referred to here as keto-limonene, keto limononaldehyde, limononic acid, and keto-limononic acid. Identified particle phase products accounted for about 60% of the observed particle mass in the initial stages of the reaction. Model sensitivity was tested and discussed with respect to effects of temperature, humidity, water uptake, and reactant concentrations. PMID- 16475338 TI - Interaction force profiles between Cryptosporidium parvum oocysts and silica surfaces. AB - The interaction force profile between single Cryptosporidium parvum oocysts and silica particles was measured in aqueous solutions using an atomic force microscope. The oocysts were immobilized during the measurements by entrapment in Millipore polycarbonate membranes with a 3 microm pore size. Experiments were performed in both NaCl and CaCl2 solutions at ionic strengths ranging from 1 to 100 mM. For both electrolytes the decay length of the repulsive force profile, obtained via the slope of a plot of the logarithm of interaction force versus separation, was found to be essentially independent of the ionic strength and always much larger than the theoretical Debye length of the system. In addition, the magnitude of the force was found to be essentially the same for both electrolytes, suggesting that the long-range repulsive forces are primarily steric in nature. Fitting the force to an expression for the steric repulsive force between two grafted brush layers yields a layer thickness of approximately 115 nm. These results support the idea that the oocysts are covered by a relatively thick layer of uncharged (or weakly charged) carbohydrates, possibly mixed with a thinner layer of charged protein. PMID- 16475341 TI - Influence of mobile air-conditioning on vehicle emissions and fuel consumption: a model approach for modern gasoline cars used in Europe. AB - The influence of air-conditioning activity on the emissions and fuel consumption of passenger cars is an important issue, since fleet penetration and use of these systems have reached a high level. Apart from the MOBILE6 study in the United States, little data is available on the impact of air-conditioning devices (A/Cs). Since weather conditions and A/C technologies both differ from those in the U. S., a test series was designed for the European setting. A fleet of six modern gasoline passenger cars was tested in different weather conditions. Separate test series were carried out for the initial cooldown and for the stationary situation of keeping the interior of the vehicle cool. As assumed, CO2 emissions and fuel consumption rise with the thermal load. This also causes a notable rise in CO and hydrocarbons (HCs). Moreover, A/Cs do not stop automatically at low ambient temperatures; if necessary, they produce dry air to demist the windscreen. A model is proposed that shows a constant load for lower temperatures and a linear trend for higher temperatures. The initial cooldown tests highlight significant differences among cars but show that A/C operation for the initial cooling of an overheated passenger compartment does not result in any extra emissions for the fleet as a whole. PMID- 16475340 TI - Vehicle specific power approach to estimating on-road NH3 emissions from light duty vehicles. AB - NH3 emissions from motor vehicles have been the subject of a number of recent studies due to their potential impact on ambient particulate matter (PM). Highly time-resolved NH3 emissions can be measured and correlated with specific driving events utilizing a tunable diode laser (TDL). It is possible to incorporate NH3 emissions with this new information into models that can be used to predict emissions inventories from vehicles. The newer generation of modal models are based on modal events, with the data collected at second-by-second time resolution, unlike the bag-based emission inventory models such as EMFAC and MOBILE. The development of an NH3 modal model is described in this paper. This represents one of the first attempts to incorporate vehicle NH3 emissions into a comprehensive emissions model. This model was used in conjunction with on-road driving profiles to estimate the emissions of SULEV, ULEV, and LEV vehicles to be 9.4 +/- 4.1, 21.8 +/- 5.2, and 34.9 +/- 6.0 mg/mi, respectively. We also implement this new NH3 model to predict and evaluate the NH3 emission inventory in the South Coast air basin (SoCAB). PMID- 16475342 TI - A quantitative assay for linking microbial community function and structure of a naphthalene-degrading microbial consortium. AB - A comprehensive culture-independent assay, called Q-FAST, was developed for concurrent identification and quantification of active microorganisms involved a specific function in a given microbial community. The development of Q-FAST was achieved by integrating the concept of stable isotope probing technique into a new quantitative fingerprinting assay called real-time-t-RFLP for microbial community structure analysis. The Q-FAST was successfully validated by using a three-member artificial microbial community containing a known naphthalene utilizing bacterium (Pseudomonas putida G7) and two nonnaphthalene-degrading bacteria (Escherichia coli and Bacillus thuringiensis). The application of Q-FAST to identify and quantify a guild of naphthalene-utilizing microorganisms in soils revealed the involvement of eight members, with six members relating to several phylogenetic groups of eubacteria (three in beta-proteobacteria, two in gamma proteobacteria, and one in genera Intrasporangium of Gram-positive bacteria) and two members showing no close phylogenetic affiliation to any known bacterial sequences deposited in GenBank. The quantity of three members belonging to beta proteobacteria accounted for 34% of total 16S rDNA copies measured from the "heavier" fraction of DNA that was contributed from the DNA of microorganisms capable of incorporating 13C-labeled naphthalene into their genetic biomarkers. The other five members composed 66% of total 16S rDNA copies of active naphthalene-utilizing populations measured. Offering a powerful tool for studying microbial ecology, Q-FAST thus opens a new avenue for deeper exploration of microbial-mediated processes, mainly the quantitative relationship between microbial diversity and microbial activity in a given environment. PMID- 16475343 TI - Analysis of atmospheric sesquiterpenes: sampling losses and mitigation of ozone interferences. AB - Atmospheric standards containing parts-per-billion levels of 14 semivolatile hydrocarbon compounds, including eight sesquiterpenes (SQTs) (longipinene, alpha copaene, isolongifolene, alpha-cedrene, trans-caryophyllene, aromadendrene, alpha humulene, delta-cadinene), two oxidized sesquiterpenoids (cisnerolidol, trans nerolidol), one biogenic ketone (geranylacetone) and three aromatic compounds (1,3,5-triisopropylbenzene, diphenylmethane, nonylbenzene), were collected onto four solid adsorbent materials at increasing ozone mixing ratios (0-100 ppbv 03) for analysis by thermodesorption-gas chromatography. Substantial sampling losses of up to >90% were found for the most reactive SQT, even at the lowest ozone level investigated of 20 ppbv. Loss rates from the ozone-SQT reaction were used to derive estimates of gas-phase ozone reaction rate constants for longipinene, alpha-copaene, isolongifolene, geranylacetone, aromadendrene, delta-cadinene, cis nerolidol, and transnerolidol. Three different ozone mitigation techniques were investigated to prevent these sampling losses. These strategies included (a) placing glass fiber filters impregnated with sodium thiosulfate (Na2S2O3) into the sampling line, (b) titration of ozone in the sampling stream with nitric oxide (NO), and (c) catalytically removing ozone with a commercially available manganese dioxide (MnO2) catalyst. All three techniques reduced ozone-mixing ratios from 100 ppbv to <0.6 ppbv at sampling flow rates of 1 L min(-1). When the Na2S2O3 filters and the NO-titration techniques were applied, SQT loss rates decreased from 25-60% to 0-5% for most SQT compounds and from >90% to approximately 10-50% for the two most reactive compounds at ozone mixing ratios of up to 100 ppbv. The commercial manganese dioxide scrubber, however, caused complete analyte losses (>98%) even at 0 ppbv ozone. These results underline the need and present applicable techniques for removal of ozone in air samples for SQT analysis by solid adsorption techniques. PMID- 16475344 TI - Seeing a deep ocean CO2 enrichment experiment in a new light: laser raman detection of dissolved CO2 in seawater. AB - We used a newly developed in situ laser Raman spectrometer (LRS) for detection of elevated levels of dissolved CO2 in seawater. The experiment was carried out at 500 m depth, 6 degrees C, to examine new protocols for detection of CO2-enriched seawater emanating from a liquid CO2 source in the ocean, and to determine current detection limits under field conditions. A system of two interconnected 5 L chambers was built, with flow between them controlled by a valve and pump system, and this unit was mounted on an ROV. The first chamber was fitted with a pH electrode and the optical probe of the LRS. In the second chamber approximately 580 mL of liquid CO2 was introduced. Dissolution of CO2 across the CO2-seawater interface then occurred, the valves were opened, and a fixed volume of low-pH/CO2-enriched seawater was transferred to the first chamber for combined pH/Raman sensing, where we estimate a mean dissolution rate of approximately 0.5 (micromol/cm2)/s. This sequence was repeated, resulting in measurement of a progressively CO2 enriched seawater sample. The rapid in-growth of CO2 was readily detected as the Fermi dyad of the dissolved state with a detection limit of approximately 10 mM with spectral acquisition times of 150 s. The detection of background levels of CO2 species in seawater (approximately 2.2 mM, dominantly HCO3-) will require an improvement in instrument sensitivity by a factor of 5-10, which could be obtained by the use of a liquid core waveguide. PMID- 16475345 TI - Heterogeneous oxidation of carbonyl sulfide on atmospheric particles and alumina. AB - Heterogeneous oxidation of carbonyl sulfide (OCS) on atmospheric particles and alumina (Al2O3) was investigated in a closed system and a flowed system using in situ diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS). At room temperature, OCS could be catalytically oxidized on the surface of atmospheric particles and Al2O3 to form gas-phase CO2 and surface sulfate (SO4(2 )), sulfite (HSO3-), and hydrogen carbonate (HCO3-) species. The real atmospheric particles were characterized using X-ray fluorescence (XRF) and the Brunauer Emmett-Teller (BET) method. As a simplified model, Al2O3 was used to study the reaction mechanism of heterogeneous oxidation of OCS. The hydrogen thiocarbonate surface (HSCO2-) species, an intermediate formed in the reaction of OCS with surface hydroxyl (OH), could only be observed on the prereduced Al2O3 sample. The experimental results also indicate that surface oxygen containing species on the atmospheric particle sample and the Al2O3 sample might be the key reactant for OCS oxidation. A reaction mechanism of heterogeneous oxidation of OCS on Al2O3 surface is discussed. PMID- 16475346 TI - Methods for accelerating nitrate reduction using zerovalent iron at near-neutral pH: effects of H2-reducing pretreatment and copper deposition. AB - Both surface treatments, H2-reducing pretreatment at 400 degrees C and the deposition of copper as a catalyst, were attempted to enhance the removal of nitrate (40 (mg N) L(-1)) using zerovalent iron in a HEPES buffered solution at a pH of between 6.5 and 7.5. After the iron surface was pretreated with hydrogen gas, the removal of the passive oxide layers that covered the iron was indicated by the decline in the oxygen fraction (energy dispersive X-ray analysis) and the overlap of the cyclic polarization curves. The reaction rate was doubled, and the lag of the early period disappeared. Then, the deposition of copper onto freshly pretreated iron promoted nitrate degradation more effectively than that onto a nonpretreated iron surface, because of the high dispersion and small size of the copper particles. An optimum of 0.25-0.5% (w/w) Cu/Fe accelerated the rate by more than six times that of the nonpretreated iron. The aged 0.5% (w/w) Cu/Fe with continual dipping in nitrate solution for 20 days completely restored its reactivity by a regeneration process with H2 reduction. Hence, these two iron surface treatments considerably promoted the removal of nitrate from near-neutral water; the reactivity of Cu/Fe was effectively recovered. PMID- 16475347 TI - Assessment of an aerosol treatment to improve air quality in a swine concentrated animal feeding operation (CAFO). AB - Poor air quality within swine concentrated animal feeding operations (CAFOs) poses a threat to workers, the surrounding community, and farm production. Accordingly, the current study was conducted to evaluate a technology for reducing air pollution including particulate matter (PM), viable bacteria, and ammonia within such a facility. The technology consists of an acid-oil-alcohol aerosol applied daily. Its effectiveness was evaluated by comparing air quality from before to after treatment and between treated and untreated sides of a barn separated by an impervious partition. On the untreated side, air quality was typical for a swine CAFO, with mean PM2.5 of 0.28 mg/m3 and PM(TOT) of 1.5 mg/m3. The treatment yielded a reduction in PM concentration of 75-90% from before to after treatment. Effectiveness increased with time, application, and particle size (40% reduction for 1 microm and 90% for >10 microm). Airborne bacteria levels (total bacteria, Enterobacteriaceae, and gram-positive cocci) decreased one logarithmic unit after treatment. In contrast, treatment had no effect on ammonia concentrations. These findings demonstrate the effectiveness of an intervention in yielding exposure and emission reductions. PMID- 16475348 TI - Impact of environmental factors on efficacy of upper-room air ultraviolet germicidal irradiation for inactivating airborne mycobacteria. AB - This study evaluated the efficacy of an upper-room air ultraviolet germicidal irradiation (UVGI) system for inactivating airborne bacteria, which irradiates the upper part of a room while minimizing radiation exposure to persons in the lower part of the room. A full-scale test room (87 m3), fitted with a UVGI system consisting of 9 louvered wall and ceiling fixtures (504 W all lamps operating) was operated at 24 and 34 degrees C, between 25 and 90% relative humidity, and at three ventilation rates. Mycobacterium parafortuitum cells were aerosolized into the room such that their numbers and physiologic state were comparable both with and without the UVGI system operating. Airborne bacteria were collected in duplicate using liquid impingers and quantified with direct epifluorescent microscopy and standard culturing assay. Performance of the UVGI system degraded significantly when the relative humidity was increased from 50% to 75-90% RH, the horizontal UV fluence rate distribution was skewed to one side compared to being evenly dispersed, and the room air temperature was stratified from hot at the ceiling to cold at the floor. The inactivation rate increased linearly with effective UV fluence rate up to 5 microW cm(-2); an increase in the fluence rate above this level did not yield a proportional increase in inactivation rate. PMID- 16475349 TI - Denitrification mechanism of NaOH in the presence of carbon. AB - We tested a mixture of NaOH and carbon and found it to react rapidly with NO gas, thus demonstrating a promising new method of flue gas denitrification (FGD). We determined the reaction products: gas phases were analyzed by GC-MS and NOx meter, and solid residues were analyzed by X-ray diffusion (XRD) and chemical analysis. The reaction process was clearly divided into two stages. In the first stage, the reaction proceeded extremely rapidly, with NO almost completely removed. The mechanism describing this stage consists of a series of reactions of NaOH and NO in the presence of carbon. The main conversion of sodium in this stage is proposed to be: NaOH-->NaNO3 (and NaNO2)-->Na2O2-Na2CO3. The pyrolysis of NaNO3 was examined and carbon was found to have the ability to reduce NOx emission during this process. In the second stage, NO reduction proceeds slowly and stabilizes at a constant value. The mechanism behind this stage is considered to be the reaction of carbon with NO in the presence of Na2-CO3 as catalyst. Quantitative study of the reaction system demonstrated that the total amount of denitrified NO is proportional to the amount of carbon and that the denitrified NO in the first stage is proportional to the amount of NaOH. PMID- 16475350 TI - A model for the effect of rhizodeposition on the fate of phenanthrene in aged contaminated soil. AB - Microcosm data were used to develop a deterministic model to describe how rhizodeposition affects the fate of phenanthrene in aged contaminated soil. Microbial mineralization and soil sequestration of 14C-phenanthrene were compared in microcosms amended weekly with phenolic-rich mulberry root extracts versus unamended controls. Mineralization was higher in the amended soils simulating the rhizosphere (57.7 +/- 0.9%) than in controls simulating bulk (unplanted) soils (53.2 +/- 0.7%) after 201 days (p < 0.05). Humin was the main soil sink for the residual 14C-label. Whereas the total 14C-label associated with humin remained constant in biologically active soils (at about 30%), it increased up to 80% after 201 days in sterile controls. The initial phenanthrene extraction with n butanol (commonly used to assess bioavailability) slightly underestimated the fraction thatwas mineralized (assessed by 14CO2 recovery). Changes in the unextractable fraction (determined by combustion in a biological oxidizer) suggested the presence of two soil sequestration domains: (1) irreversibly bound residue, and (2) an intermediate transition phase that is unextractable by solvents at a given point in time but could become bioavailable due to physicochemical or biological transformations of the binding matrix. The fate of phenanthrene was accurately modeled by considering the transfer of the 14C label between different soil compartments as first-order kinetic processes. Model simulations suggested that the system was approaching a stable end-point after 201 days of simulated rhizoremediation, and corroborated that microorganisms have a significant impact on the fate of phenanthrene in soil. PMID- 16475351 TI - Mineral CO2 sequestration by steel slag carbonation. AB - Mineral CO2 sequestration, i.e., carbonation of alkaline silicate Ca/Mg minerals, analogous to natural weathering processes, is a possible technology for the reduction of carbon dioxide emissions to the atmosphere. In this paper, alkaline Ca-rich industrial residues are presented as a possible feedstock for mineral CO2 sequestration. These materials are cheap, available near large point sources of CO2, and tend to react relatively rapidly with CO2 due to their chemical instability. Ground steel slag was carbonated in aqueous suspensions to study its reaction mechanisms. Process variables, such as particle size, temperature, carbon dioxide pressure, and reaction time, were systematically varied, and their influence on the carbonation rate was investigated. The maximum carbonation degree reached was 74% of the Ca content in 30 min at 19 bar CO2 pressure, 100 degrees C, and a particle size of <38 microm. The two most important factors determining the reaction rate are particle size (<2 mm to <38 microm) and reaction temperature (25-225 degrees C). The carbonation reaction was found to occur in two steps: (1) leaching of calcium from the steel slag particles into the solution; (2) precipitation of calcite on the surface of these particles. The first step and, more in particular, the diffusion of calcium through the solid matrix toward the surface appeared to be the rate-determining reaction step. The Ca diffusion was found to be hindered by the formation of a CaCO3-coating and a Ca-depleted silicate zone during the carbonation process. Research on further enhancement of the reaction rate, which would contribute to the development of a cost-effective CO2-sequestration process, should focus particularly on this mechanism. PMID- 16475352 TI - Increasing Fe0-mediated HMX destruction in highly contaminated soil with didecyldimethylammonium bromide surfactant. AB - Mixtures of energetic compounds pose a remediation problem for munitions contaminated soil. Although treatment with zerovalent iron (Fe0) can be effective, RDX and TNT are more readily destroyed than HMX. Adding didecyldimethylammonium bromide (didecyl) at 2% w/v with 3% (w/v) Fe0 to a 20% slurry of Los Alamos National Laboratory soil containing solid-phase HMX (45 000 mg/kg) resulted in >80% destruction within 6 days. Because the HMX concentration did not increase in solution and the didecyl equilibrium concentration was well below the critical micelle concentration, we conclude thatthe solution primarily contained didecyl monomers. The adsorption isotherm for didecyl on iron is consistent with electrostatic adsorption of monomers and some hydrophobic partitioning at low equilibrium concentrations. Fe0 pretreated with didecyl was superior to Fe0 alone or mixed with didecyl in removing HMX from solution, but it was less effective than Fe0 + didecyl when solid-phase HMX was present. Reseeding HMX to mimic dissolution indicated an initial high reactivity of didecyl pretreated Fe0, but the reaction slowed with each HMX addition. In contrast, reaction rates were lower but reactivity was maintained when Fe0 and didecyl were added together and didecyl was included in fresh HMX solutions. Destruction of solid-phase HMX requires low didecyl concentrations in solution so that hydrophobic patches are maintained on the iron surface. PMID- 16475353 TI - Evaluation of mixed valent iron oxides as reactive adsorbents for arsenic removal. AB - The objective of this research was to determine if Fe(II)-bearing iron oxides generate ferric hydroxides at sufficient rates for removing low levels of arsenic in packed-bed reactors, while at the same time avoiding excessive oxide production that contributes to bed clogging in oxygenated waters. Column experiments were performed to determine the effectiveness of three media for arsenic removal over a range in empty bed contact times, influent arsenic concentrations, dissolved oxygen (DO) levels, and solution pH values. Corrosion rates of the media as a function of the water composition were determined using batch and electrochemical methods. Rates of arsenic removal were first order in the As(V) concentration and were greater for media with higher corrosion rates. As(V) removal increased with increasing DO levels primarily due to faster oxidation of the Fe2+ released by media corrosion. To obtain measurable amounts of arsenic removal in 15 mM NaCl electrolyte solutions containing 50 microg/L As(V), the rate of Fe2+ released by the media needed to be at least 15 times greater than the As(V) feed rate into the column. In waters containing 30 mg/L of silica and 50 microg/L of As(V), measurable amounts of arsenic removal were obtained only for Fe2+ release rates that were at least 200 times greater than the As(V) feed rate. Although all columns showed losses in hydraulic conductivity overthe course of 90 days of operation, the conductivity values remained high, and the losses could be reversed by backwashing the media. The reaction products produced by the media in domestic tap water had average As-to-Fe ratios that were approximately 25% higher than those for a commercially available adsorbent. PMID- 16475354 TI - Oxidation mechanism of As(III) in the UV/TiO2 system: evidence for a direct hole oxidation mechanism. AB - Although it is well-known that As(III) is oxidized to As(V) in the UV/TiO2 system, the main oxidant for that reaction is still not clear. Accordingly, the present study aims at reinvestigating the TiO2-photocatalyzed oxidation mechanism of As(III). We performed a series of As(II) oxidation experiments by using UV C/H2O2 and UV-A/TiO2, focusing on the effects of competing compounds. The experiment with UV-C/H2O2 indicated that HO2*/O2-* is not an effective oxidant of As(III) in the homogeneous phase. The effects of oxalate, formate, and Cu(II) on the photocatalytic oxidation of As(III) contradicted the controversial hypothesis that HO2*/ O2-* is the main oxidant of As(III) in the UV/TiO2 system. The effect of As(III) on the TiO2-photocatalyzed oxidations of benzoate, terephthalate, and formate was also incompatible with the superoxide-based As(II) oxidation mechanism. Instead, the experimental observations implied that OH* and/or the positive hole are largely responsible forthe oxidation of As(III) in the UV/TiO2 system. To determine which species plays a more significant role, the effects of methanol and iodide were tested. Since excess methanol did not retard the oxidation rate of As(III), OH* seems not to be the main oxidant. Therefore, the best rationale regarding the oxidation mechanism of As(III) in the UV/TiO2 system seems to be the direct electron transfer between As(III) and positive holes. Only with this mechanism, it was possible to explain the data of this study. Besides the mechanistic aspect, an application method for this technology was sought. The usage of UV/TiO2 for oxidizing As(II) requires a posttreatment in which both As(V) and TiO2 should be removed from water. For this objective, we applied FeCl3 and AIK(SO4)2 as coagulants, and the result implied that the combined usage of TiO2 and coagulation might be a feasible solution to treat arsenic contamination around the world. PMID- 16475355 TI - UV photolytic mechanism of N-nitrosodimethylamine in water: roles of dissolved oxygen and solution pH. AB - The photolytic mechanism of N-nitrosodimethylamine (NDMA) in aqueous solution was investigated under ultraviolet (UV) irradiation at 253.7 nm, by determining the quantum yield of NDMA photolysis and analyzing the photolysis products in the pH range of 2-11 under O2 or N2 saturation. Several important photolytic behaviors of NDMA photolysis regarding the roles of dissolved oxygen and solution pH were newly observed, and novel photolytic mechanisms were proposed based on these observations. First, the presence of dissolved oxygen clearly increased the quantum yield of the NDMA photolysis, and promoted the photolytic pathway leading to the formation of methylamine and nitrate. The direct oxidation of photoexcited NDMA by dissolved oxygen (photooxidation) was suggested as a novel photolytic pathway to explain the observed effect of dissolved oxygen. This photooxidation mechanism was supported by the detection of superoxide radicals during the photolysis of NDMA under O2 saturation. Second, the dependence of the quantum yield of NDMA photolysis and the yields of the known photolysis products (methylamine, dimethylamine, nitrite, and nitrate) on the solution pH were different under O2 and N2 saturation. N-Methylformamide was observed as the novel photolysis product of NDMA in the neutral pH region under N2 saturation. These pH dependent photolytic behaviors of NDMA were successfully explained by the interaction of the photoexcited NDMA with a proton, and the pH-dependent secondary reactions between the primary photolysis products. PMID- 16475356 TI - Influence of O2 and H2O on carbothermal reduction of SO2 by oil-sand fluid coke. AB - To develop a new process for removing high-concentration SO2 from industrial flue gases, the carbothermal reduction of SO2 by oil-sand fluid coke at 700 degrees C was investigated by varying the inlet concentration of either O2 or H2O. Concentrations of O2 and H2O ranged from 0 to 20% and from 0 to 30%, respectively, in a stream of SO2 (18%) with the balance helium. Addition of O2 and H2O was found to enhance SO2 reduction. The enhancement was attributed to the reducing gases, CO and H2, produced by solid-gas reactions between carbon and O2 or H2O. The effects of O2 and H2O on sulfur yield, however, were bifacial: adding O2 and/or H2O increased the sulfur yield when SO2 conversion was incomplete, otherwise, it decreased the sulfur yield through the formation of sulfides such as H2S. The results of a thermodynamic analysis were in a good agreementwith the experimental results, suggesting that gas-solid reactions were slow enough to allow gas-phase equilibrium. This study indicates that carbon, such as oil-sand fluid coke, can be utilized to remove SO2 in flue gases containing O2/H2O and to convert it to elemental sulfur. PMID- 16475357 TI - SOx removal by calcined MgAlFe hydrotalcite-like materials: effect of the chemical composition and the cerium incorporation method. AB - Sulfur oxides are one of the most hazardous atmospheric pollutants since they contribute directly to acid rain formation. Consequently, stringent environmental regulations limit atmospheric SOx emissions, motivating research on efficient ways to reduce them. To supply an alternative to reduce these emissions in fluid catalytic cracking units, this study discloses efficient SOx reducing materials based on calcined MgAlFe hydrotalcite-like compounds (HT's). Thus, HT materials were synthesized by several methods including cerium addition. The adsorption of SO2 was carried out by contacting the calcined solid with a mixture of SO2 (1%) in air at 650 degrees C. It was demonstrated that the isomorphic incorporation of iron increased its reduction capability which was reflected in higher reduction rates and metal sulfate reduction grade at 550 degrees C. Moreover, when cerium was present in the iron-containing materials the saturation rate was improved, because cerium oxide promotes the oxidation of SO2 to SO3. The way cerium is incorporated influences the SO2 adsorption capacity. PMID- 16475358 TI - Amination of n-hexanol in supercritical water. AB - The amination of 1-n-hexanol followed by amidation was carried out in supercritical water at 380, 400, and 420 degrees C and water densities of 0.1, 0.3, and 0.5 g/cm3. The replacement of the hydroxyl group with the amino group was found to occur in 1-n-hexanol using ammonium acetate in supercritical water without the addition of a metal or an acid catalyst. The yield of the final product, N-n-hexylacetamide, increased by increasing the reaction temperature, water density, and the amount of ammonium acetate. The yield and the selectivity of N-n-hexylacetamide were 78.5% and 87.5%, respectively, in supercritical water at 400 degrees C, 0.5 g/cm3, for 10 min. PMID- 16475359 TI - Reduction of octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine by zerovalent iron: product distribution. AB - RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine) and HMX (octahydro-1,3,5,7 tetranitro-1,3,5,7-tetrazocine) are cyclic nitramines ((CH2NNO2)n; n = 3 or 4, respectively) widely used as energetic chemicals. Their extensive use led to wide environmental contamination. In contrast to RDX, HMX tends to accumulate in soils due to its unique recalcitrance. In the present study, we investigated the potential of zerovalent iron (ZVI) to transform HMX under anoxic conditions. HMX underwent a rapid transformation when added in well-mixed anoxic ZVI-H2O batch systems to ultimately produce formaldehyde (HCHO), ammonium (NH4+), hydrazine (NH2NH2), and nitrous oxide (N2O). Time course experiments showed that the mechanism of HMX transformation occurred through at least two initial reactions. One reaction involved the sequential reduction of N-NO2 groups to the five nitroso products (1NO-HMX, cis-2NO-HMX, trans-2NO-HMX, 3NO-HMX, and 4NO-HMX). Another implied ring cleavage from either HMX or 1NO-HMX as demonstrated by the observation of methylenedinitramine (NH(NO2)CH2NH(NO2)) and another intermediate that was tentatively identified as (NH(NO2)CH2N(NO)CH2NH-(NO2)) or its isomer (NH(NO)CH2N(NO2)CH2NH(NO2)). This is the first study that demonstrates transformation of HMX by ZVI to significant amounts of NH2NH2 and HCHO. Both toxic products seemed to persist under reductive conditions, thereby suggesting that the ultimate fate of these chemicals, particularly hydrazine, should be understood prior to using zerovalent iron to remediate cyclic nitramines. PMID- 16475360 TI - Comparison of the effects of sonolysis and gamma-radiolysis on dissolved organic matter. AB - The effects of 640 kHz sonolysis and 60Co gamma-radiolysis on dissolved organic matter (DOM) were compared through UV/ vis absorption spectrometric, dissolved organic carbon concentration ([DOC]), and potentiometric titration analyses. A reverse-phase chromatographic technique was used to compare changes in the DOM hydrophobicity distribution, and a size exclusion chromatographic technique with inline UV-A absorbance, fluorescence, and [DOC] detectors was used to compare changes in the DOM molecular weight distribution. Whereas upon radiolysis major decreases in absorbance and [DOC] were induced and near-total DOC removal was achieved, upon sonolysis there were major decreases in UV/vis absorbance but only minor decreases in [DOC], and a substantial quantity of hydrophilic nonchromophoric material remained in solution. In radiolysis, hydrophilic and hydrophobic DOM solution components were exposed to equal hydroxyl radical (*OH) concentrations. However, in sonolysis, hydrophobic DOM components were exposed to more elevated *OH concentrations than the hydrophilic components and consequently had enhanced rates of degradation. Sonolysis may be of interest in the design of advanced oxidation processes in which the selective elimination of hydrophobic solution components, such as hydrophobic organic contaminants and hydrophobic DOM domains into which they partition, is desired. PMID- 16475361 TI - Permeable membranes containing crystalline silicotitanate as model barriers for cesium ion. AB - In diaphragm cell experiments, a permeable model reactive barrier for the containment of cesium is tested. Primary targets for cesium containment are former plutonium processing sites (e.g., Hanford, WA and Savannah River, SC), which are currently contaminated with cesium-137. Adding up to 10 wt % crystalline silicotitanate, a sacrificial reagent, to poly(vinyl alcohol) films increases the time before cesium can cross the film by a factor of 30. The increased lag times are consistent with theories developed for this type of reactive membrane. Theory also correctly predicts the effects of cesium concentration and membrane thickness on membrane performance. Because the relative improvements of the model barrier are expected to be independent of the polymer used, these increased lags should hold for less permeable polymers that are more resistant to radiation, although these polymers have not been tested. PMID- 16475362 TI - Adsorption of hydrogen sulfide onto activated carbon fibers: effect of pore structure and surface chemistry. AB - To understand the nature of H2S adsorption onto carbon surfaces under dry and anoxic conditions, the effects of carbon pore structure and surface chemistry were studied using activated carbon fibers (ACFs) with different pore structures and surface areas. Surface pretreatments, including oxidation and heattreatment, were conducted before adsorption/desorption tests in a fixed-bed reactor. Raw ACFs with higher surface area showed greater adsorption and retention of sulfur, and heat treatment further enhanced adsorption and retention of sulfur. The retained amount of hydrogen sulfide correlated well with the amount of basic functional groups on the carbon surface, while the desorbed amount reflected the effect of pore structure. Temperature-programmed desorption (TPD) and thermal gravimetric analysis (TGA) showed that the retained sulfurous compounds were strongly bonded to the carbon surface. In addition, surface chemistry of the sorbent might determine the predominant form of adsorbate on the surface. PMID- 16475363 TI - Life cycle assessment of switchgrass- and corn stover-derived ethanol-fueled automobiles. AB - Utilizing domestically produced cellulose-derived ethanol for the light-duty vehicle fleet can potentially improve the environmental performance and sustainability of the transport and energy sectors of the economy. A life cycle assessment model was developed to examine environmental implications of the production and use of ethanol in automobiles in Ontario, Canada. The results were compared to those of low-sulfur reformulated gasoline (RFG) in a functionally equivalent automobile. Two time frames were evaluated, one near-term (2010), which examines converting a dedicated energy crop (switchgrass) and an agricultural residue (corn stover) to ethanol; and one midterm (2020), which assumes technological improvements in the switchgrass-derived ethanol life cycle. Near-term results show that, compared to a RFG automobile, life cycle greenhouse gas (GHG) emissions are 57% lower for an E85-fueled automobile derived from switchgrass and 65% lower for ethanol from corn stover, on a grams of CO2 equivalent per kilometer basis. Corn stover ethanol exhibits slightly lower life cycle GHG emissions, primarily due to sharing emissions with grain production. Through projected improvements in crop and ethanol yields, results for the mid term scenario show that GHG emissions could be 25-35% lower than those in 2010 and that, even with anticipated improvements in RFG automobiles, E85 automobiles could still achieve up to 70% lower GHG emissions across the life cycle. PMID- 16475364 TI - Flow of natural versus economic capital in industrial supply networks and its implications to sustainability. AB - Appreciating the reliance of industrial networks on natural capital is a necessary step toward their sustainable design and operation. However, most contemporary accounting techniques, including engineering economics, life cycle assessment, and full cost accounting, fail in this regard, as they take natural capital for granted and concentrate mainly on the economic aspects and emissions. The recently developed "thermodynamic input-output analysis" (TIOA) includes the contribution of ecological goods, ecosystem services, human resources, and impact of emissions in an economic input-output model. This paper uses TIOA to determine the throughputs of natural and economic capitals along industrial supply networks. The ratios of natural to economic capitals of economic sectors reveals a hierarchical organization of the U.S. economy wherein basic infrastructure industries are at the bottom and specialized value-added industries constitute the top. These results provide novel insight into the reliance of specific industrial sectors and supply chains on natural capital and the corresponding economic throughput. Such insight is useful for understanding the implications of corporate restructuring on industrial sustainability metrics and of outsourcing of business activities on outsourcer, outsourcee, and global sustainability. These implications are discussed from the standpoints of weak and strong sustainability paradigms. The calculated ratios can also be used for hybrid thermodynamic life cycle assessment. PMID- 16475365 TI - Parametric analysis of environmental performance of reused/recycled packaging. AB - Annual environmental performance of packaging products which are reused at least once per year is analyzed with respect to three specific criteria: (1) waste quantities, (2) virgin material demand and resource depletion, and (3) environmental impacts from manufacturing. Packaging flow performance is assessed via a combined reuse/ recycle rate index expressed solely in terms of two dimensionless parameters: the conventional recycling rate and the mean number of reuse trips. Quantitative expressions describe the effect of the following physical quantities: annual reuse frequency, lifetime, maximum number of reuse trips, amount of packaging present in the market, annual production plus net trade imports, recycle rate of consumer discard, reuse rate and consumer discard. The results may serve for packaging monitoring and assessment of alternative packaging systems and for setting more efficient environmental policy targets in terms of the reuse/recycle rate. PMID- 16475366 TI - Aquatic toxicity due to residential use of pyrethroid insecticides. AB - Pyrethroids are the active ingredients in most insecticides available to consumers for residential use in the United States. Yet despite their dominance in the marketplace, there has been no attempt to analyze for most of these compounds in watercourses draining residential areas. Roseville, California was selected as a typical suburban development, and several creeks that drain subdivisions of single-family homes were examined. Nearly all creek sediments collected caused toxicity in laboratory exposures to an aquatic species, the amphipod Hyalella azteca, and about half the samples caused nearly complete mortality. This same species was also found as a resident in the system, but its presence was limited to areas where residential influence was least. The pyrethroid bifenthrin is implicated as the primary cause of the toxicity, with additional contributions to toxicity from the pyrethroids cyfluthrin and cypermethrin. The dominant sources of these pyrethroids are structural pest control by professional applicators and/ or homeowner use of insecticides, particularly lawn care products. The suburbs of Roseville are unlikely to be unique, and similar sediment quality degradation is likely in other suburban areas, particularly in dry regions where landscape irrigation can dominate seasonal flow in some water bodies. PMID- 16475367 TI - Isolation of the retinoblastoma cDNA from the marine flatfish dab (Limanda limanda) and evidence of mutational alterations in liver tumors. AB - We have isolated a dab (Limanda limanda) homologue of the human retinoblastoma (Rb) tumor suppressor gene. The L. limanda partial Rb cDNA encodes a partial predicted protein of 753 amino acids. DNA sequence analysis with other vertebrate Rb sequences demonstrates that the L. limanda Rb cDNA is highly conserved in regions of functional importance. The sequence reported herein, combined with the high degree of conservation observed in critical domains, has also facilitated an investigation of the molecular etiology of environmentally induced liver tumor samples in a feral fish species. Mutational alterations were detected in liver adenoma samples, also in apparently "normal" regions of liver samples dissected from fish displaying adenoma, but not in normal liver samples from otherwise healthy feral fish. These results are the first reporting the appearance of Rb mutations in wild-caught fish and suggest that the molecular etiology of fish cancer appears to involve Rb-implicated tumorigenesis. The ecotoxicological relevance of the Rb mutations in feral fish liver tumors, in terms of future genome instability and possible development of a genotoxicity biomarker, is discussed. PMID- 16475369 TI - Technologies that alter the way we practice. PMID- 16475368 TI - Modulation of brain steroidogenesis by affecting transcriptional changes of steroidogenic acute regulatory (StAR) protein and cholesterol side chain cleavage (P450scc) in juvenile Atlantic salmon (Salmo salar) is a novel aspect of nonylphenol toxicity. AB - Gene expression patterns for key brain steroidogenic (StAR, P450scc, CYP11beta) and xenobiotic- and steroid-metabolizing enzymes (CYP1A1 and CYP3A) have been investigated in waterborne nonylphenol (5, 15, and 50 microg/ L) treated juvenile Atlantic salmon (Salmo salar), in addition to carrier vehicle (ethanol) exposed fish, sampled at different time intervals (0, 3, and 7 days) after exposure. Gene expression patterns were studied using the quantitative polymerase chain reaction (real-time PCR). Treatment of juvenile salmon with nonylphenol caused significant induction of steroidogenic acute regulatory (StAR) protein mRNA at day 7 postexposure in the group receiving 15 microg of nonylphenol/L. P450scc was first induced in the group treated with 5 microg of nonylphenol/L at day 7; thereafter, an apparent nonylphenol-concentration-dependent decrease in P450scc mRNA was observed. CYP11beta mRNA was significantly induced at day 3 after exposure to 5 betag of nonylphenol/L; thereafter, CYP11beta mRNA levels were inhibited below control levels in the 15 and 50 microg of nonylphenol/L groups at day 3. At day 7, significant induction of CYP11beta mRNA was observed only in the group exposed to 15 microg of nonylphenol/L. For CYP1A1 mRNA, apparent nonylphenol concentration-dependent decreases were observed at day 7 postexposure. CYP3A mRNA was significantly induced by all nonylphenol exposure concentrations at day 7. When exposed groups were compared, CYP3A transcript was significantly induced between 5 and 15 microg of nonylphenol/ L, and decreased between 15 and 50 microg of nonylphenol/ L. The ethanol control showed a significant reduction of CYP3A mRNA at day 3 postexposure. The present study has demonstrated variations in three key steroidogenic proteins and xenobiotic- and steroid-metabolizing CYP isoenzyme gene transcripts in the brain of nonylphenol-exposed juvenile salmon. Therefore, the present study represents a novel aspect of neuroendocrine effects of nonylphenol in fish not previously demonstrated and should be studied in more detail. PMID- 16475370 TI - A new shading concept based on natural tooth color applied to direct composite restorations. AB - OBJECTIVE: Patient demands have prompted manufacturers to improve intrinsic optical properties of resin composites and clinicians to refine application procedures. The aim of this study is to present a shading concept based on colorimetric L*a*b* and contrast ratio data of human dentin and enamel. METHOD AND MATERIALS: Extracted teeth of the A and B Vita shade groups (n = 8 per group) were sectioned according to 2 different planes to measure specific color (using the CIE L*a*b* system) and opacity (contrast ratio). Standardized samples of enamel and dentin shades of a new composite system (Miris, Coltene Whaledent) were submitted to the same colorimetric evaluation for comparison with natural tissues. RESULTS: Comparison of teeth from the Vita groups A and B having the same chroma showed limited variations regarding a* (green to red) and b* (blue to yellow) values; the only significant variation was the increasing b* values (yellow) with increasing chroma (A1 to A4 and B1 to B3). As for dentin contrast ratio, limited differences were reported, while enamel proved to increase in translucency with age (reduced contrast ratio). CONCLUSION: These data served as the foundation of the so-called natural layering concept, which makes use of 2 basic composite masses (dentin and enamel) that optically mimic natural tissues. This concept allows for simplified clinical application and layering of composite, as it uses only 1 universal dentin hue with several chroma levels and 3 enamel types for young, adult, and old patients, each exhibiting specific tints and translucency levels. PMID- 16475371 TI - Bisphosphonate therapy for bone loss in patients with osteoporosis and periodontal disease: clinical perspectives and review of the literature. AB - As the baby boomer generation in the United States ages, more patients are using bisphosphonates for systemic bone diseases like osteoporosis. Because of their ability to inhibit bone resorption and osteoclastic activity, bisphosphonates may also be beneficial in modulating host response for periodontal disease management. This literature review examines the mechanism of action for bisphosphonates and their uses in treating periodontal disease. The dental profession should continue to evaluate this class of drugs and to closely monitor patients who are on bisphosphonate therapy. PMID- 16475372 TI - Microleakage of Class V resin composites placed using self-etching resins: effect of prior enamel etching. AB - OBJECTIVE: The purpose of this study was to compare the seal along dentin and enamel margins produced by 3 self-etching resin bonding systems to that of an etch-and-rinse adhesive system. Potential improvement in adaptation along enamel margins through the experimental use of conventional enamel etching prior to application of the self-etching adhesives was also evaluated. METHOD AND MATERIALS: Class V resin composite restorations were placed in prepared cavities in extracted third molars using 3 self-etching dentin adhesive systems: Adper Prompt L-Pop (3M Espe), iBond GI (Heraeus Kulzer), and Tyrian SPE (Bisco), and an etch-and rinse adhesive, Adper ScotchBond Multi-Purpose (3M Espe). A group of teeth also underwent conventional enamel etching prior to use of the self-etching products. The restored teeth were thermocycled, subjected to a dye challenge, and sectioned. The sections were scored using an ordinal leakage scale (n = 20). Ranked data were analyzed using a 2-way analysis of variance and Tukey multiple comparison. RESULTS: When used as directed, there were no significant differences along dentin margins for any of the adhesives, but Adper Prompt and iBond demonstrated significantly greater leakage than the etch-and-rinse product along enamel margins. The incidence of enamel margin leakage decreased significantly for the same 2 products when enamel etching preceded use of the resin. Enamel etching increased the leakage along dentin margins for all 3 self-etching products, but not significantly. CONCLUSION: Considerable improvement of all classes of dentin-adhesive resin systems is still needed. Practitioners should await further clinical trials before adopting use of self-etching resins, especially for large occlusal restorations. PMID- 16475373 TI - Number of in-office light-activated bleaching treatments needed to achieve patient satisfaction. AB - OBJECTIVE: In-office bleaching has become very popular for patients desiring immediate results. This report discusses the efficacy of in-office bleaching using high-intensity xenon halogen tungsten light on 73 patients in a private practice over 6 months. METHOD AND MATERIALS: Thirty-five percent hydrogen peroxide gel was applied to the tooth surfaces, and then both arches were illuminated with the LumaArch unit light for 8 minutes. The entire procedure was completed 3 times for a total application time of 24 minutes. Thereafter, patients returned every 2 weeks for another 24-minute in-office treatment until the patient was satisfied or did not want additional treatment. After completion of bleaching, patients were classified into groups according to the number of treatments they received. When requested, home bleaching was used as a follow-up treatment. The degree of color change and color relapse were evaluated at baseline and immediately, 2 weeks, and 6 months after bleaching. Also evaluated was the number of patients that requested home bleaching. RESULTS: Of the 73 patients who received 1 to 4 in-office bleaching procedures, 58 were satisfied. Twenty-seven patients requested home bleaching. The average color change per appointment was 2.1 to 3.7 units on a 16-scale Vita Classic shade guide. Color relapse was more noticeable at the 2-week interval than at the 6-month postbleaching evaluation. CONCLUSION: In-office bleaching may be an alternative for patients who do not like home bleaching. In-office treatment may achieve satisfactory results, but often more than 1 visit is necessary to achieve patient satisfaction. PMID- 16475374 TI - A review of dens invaginatus (dens in dente) in permanent and primary teeth: report of a case in a microdontic maxillary lateral incisor. AB - An unusual presentation of dens invaginatus affecting a microdontic permanent lateral incisor in a 19-year-old female patient is reported and pertinent literature reviewed. The patient was referred to the oral and maxillofacial radiology clinic of the authors' institution for radiographic examination of her maxillary teeth. Dens invaginatus was identified on the maxillary right lateral incisor in the anterior periapical radiographs. The tooth was microdontic and clinically simulated a primary tooth. Dens invaginatus occurs rarely in primary dentition although it is fairly common in permanent teeth. Only 4 instances of dens invaginatus affecting the primary dentition have been reported in the literature. The etiology, pathophysiology, association with other dental anomalies, as well as management aspects of this common anomaly, are discussed. PMID- 16475375 TI - Utilization of mandibular tori for alveolar ridge augmentation and maxillary sinus lifting: a case report. AB - This article reports the utilization of mandibular tori as an alternative source of autogenous bone for ridge augmentation and maxillary sinus lifting to facilitate ideal implant placement. A patient presenting bilateral mandibular tori required replacement of a missing maxillary molar and a mandibular premolar. Both areas showed ridge deficiency that required bone augmentation before implant placement. Mandibular tori were used for horizontal augmentation of the mandibular alveolar ridge and vertical augmentation of the maxillary ridge by elevation of the maxillary sinus. Adequate new bone formation was noted in both areas 6 months after grafting. Bone formed in the grafted areas showed comparable clinical features to those of native bone. Implants were successfully placed and loaded, restoring esthetics and function. Within the limitations of this observation, mandibular tori can be successfully used for ridge augmentation and sinus lifting. However, further controlled studies are needed to determine the overall benefit of this source of autogenous bone. PMID- 16475376 TI - A prospective clinical study of indirect and direct composite and ceramic inlays: ten-year results. AB - OBJECTIVE: The aim of this investigation was to evaluate the clinical performance of 4 types of tooth-colored inlays. METHOD AND MATERIALS: Fifteen direct ceramic inlays (Cerec Cos 2.0), 15 direct composite inlays (Brilliant DI, Coltene), 14 indirect ceramic inlays (Vita Dur N), and 14 indirect composite inlays (Estilux, Kulzer) were placed in 37 patients, according to manufacturer instructions. The inlays were evaluated 1 week (baseline), 6 months, and 1, 3, 4, 5, and 10 years after cementation (modified California Dental Association Quality Evaluation System). The ratings were compared using the chi-square test. For comparing the survival rates among the 4 types of inlays, a life-table analysis was done, followed by a log-rank test. RESULTS: Three Vita Dur N inlays were replaced after 1, 4, and 8.5 years; 3 Cerec inlays were replaced after 4.5, 8.5, and 9.5 years; and 3 Brilliant DI inlays needed replacement after 1, 5, and 6.5 years, all because of secondary caries or fractures. Three inlays (Estilux) were replaced because of persisting hypersensitivity or pulpal damage. Six inlays (3 Vita Dur N, 1 Estilux, and 2 Brilliant DI) were repaired for minor fractures. During the observation period, the surface texture of Vita Dur N inlays became significantly rougher. About 80% of the inlays, including repaired inlays, were in function after 10 years. CONCLUSION: After 10 years of observation, survival of the 4 types of tooth-colored inlays was similar and considered clinically acceptable. The survival rates were within the range of survival for direct composite restorations. PMID- 16475378 TI - Clinical images in oral medicine. Amalgam tattoos. PMID- 16475377 TI - Fluoride release from esthetic restorative materials with and without adhesive. AB - OBJECTIVE: The purpose of this in vitro study was to compare the effect of surface coating containing fluoride on patterns and amounts of fluoride release from 5 esthetic restorative materials. METHOD AND MATERIALS: Twelve cylinders of each material were prepared in a plexiglass mold. The experimental groups (n = 6) were coated with 1 layer of fluoridated adhesive resin, Prompt L-Pop, while the control groups (n = 6) remained uncoated. Fluoride release into deionized water was measured on days 1, 2, 3, 7, 14, 21, and 28 using an ion analyzer. Results were evaluated statistically using repeated measure analysis of variance and Duncan test. RESULTS: In the uncoated materials, the highest fluoride-releasing material was a resin composite Ariston pHc (140.468 +/- 9.80) followed by Photac fil (101.200 +/- 5.56), Ketac-fil (91.098 + 4.69), Hytac Aplitip (5.122 +/- 1.00), and Ecusit (0.310 +/- 0.18). Material-coating interaction was found to be significant, but the effect of surface coating was different for each material. Among the coated materials, conventional glass-ionomer cement Ketac-fil released the highest amount of fluoride (93.326 +/- 10.86), followed by Photac-fil (83.666 +/- 4.72), Ariston pHc (53.862 +/- 7.90), Hytac Aplitip (14.634 +/- 2.35), and Ecusit (1.355 +/- 0.29). CONCLUSION: Application of fluoridated adhesive affected fluoride release from each material in varying magnitudes. PMID- 16475380 TI - A turnover time shows wide variations. PMID- 16475379 TI - OR managers' role as gatekeepers for MD credentialing, privileging. PMID- 16475381 TI - Turnover strategies from better performers. PMID- 16475382 TI - OR of future reports improved throughput. PMID- 16475383 TI - Project reduces total joint turnover 50%. PMID- 16475384 TI - Scoring fire risk for surgical patients. PMID- 16475385 TI - Are your elderly patients safe to go home? PMID- 16475386 TI - Coding correctly for full reimbursement. PMID- 16475387 TI - Daylight matters. PMID- 16475388 TI - When things go bump in the night. PMID- 16475389 TI - Making the diagnosis. PMID- 16475390 TI - The Nighthawks. PMID- 16475391 TI - A good night's sleep. PMID- 16475392 TI - The dark side of tanning. PMID- 16475393 TI - Hot topic. PMID- 16475394 TI - A sleeper of a subject. PMID- 16475395 TI - A challenge to improve quality of care. PMID- 16475396 TI - A conversation with our board chair. PMID- 16475398 TI - Father time. PMID- 16475397 TI - I'm sorry. PMID- 16475399 TI - The saddest season. PMID- 16475400 TI - The role of vitamin D deficiency in osteoporosis and fractures. AB - Vitamin D deficiency is a major risk factor for accelerated bone loss, which contributes to morbidity in older adults. It is easy to diagnose, and treatment is safe and cost-effective. This article reviews the amount of vitamin D needed by certain populations in order to help prevent osteoporosis and fractures associated with the disease. It also summarizes the results of a study involving 81 hip-fracture patients that found 80% had abnormally low levels of 25 hydroxyvitamin D. PMID- 16475401 TI - Top 10 vitamin D myths. AB - A growing body of evidence points to the tremendous importance of vitamin D in maintaining health. Yet misconceptions about the need for this vitamin and the clinical care of vitamin D-deficient patients remain. This article deconstructs 10 prevailing myths and addresses such issues as symptoms and clinical indicators of vitamin D deficiency; the various sources of vitamin D, including diet, sun exposure, and supplementation; current laboratory standards for assessing vitamin D levels, and the prevalence of deficiency. PMID- 16475402 TI - Battling the bulge. PMID- 16475403 TI - Treadmills, tracks, and hockey sticks. PMID- 16475404 TI - Fat-finding mission. PMID- 16475405 TI - Stimulating idea. PMID- 16475406 TI - Reshaping the body and mind. PMID- 16475407 TI - The real bottom line. PMID- 16475408 TI - Not too high, not too low. PMID- 16475409 TI - The straight shooter. PMID- 16475410 TI - No longer a sideshow. PMID- 16475411 TI - Confessions of a 600-pound patient. PMID- 16475412 TI - How to evaluate and treat obesity. AB - Obesity has been recognized as a preventable and treatable disease by medical experts in Minnesota and elsewhere, and there is a growing evidence base showing the efficacy of various clinical strategies for its management. It has been shown that losing as little as 5% to 10% of body weight can help patients improve their glycemic levels, blood pressure, and lipid levels. This article discusses how to recognize obesity, identify patients who are at the greatest risk from complications of the disease, and determine which patients might respond best to certain treatments including behavior modification, diet, exercise, medication, and surgery. PMID- 16475413 TI - Minnesota physicians' attitudes and behaviors regarding weight. AB - The increasing girth of Americans is a major public health issue that also has a substantial economic impact. Consequently, physicians are being encouraged to diagnose and treat their overweight and obese patients. This article reports on the results of a study designed to gain insight into Minnesota physicians' own health habits, their efforts to recognize unhealthy weight gain and obesity among patients, and their attempts to communicate with their patients about their weight. PMID- 16475414 TI - Role of nonexercise activity thermogenesis (NEAT) in obesity. AB - Obesity develops when there is an imbalance between energy intake and energy expenditure, which can vary daily within and among individuals. High levels of energy intake and low levels of energy expenditure contribute to obesity, both together and independently. Energy expenditure from exercise associated with formal programs is encouraged for health and weight loss, but most individuals get very little formal exercise. Nonexercise activity thermogenesis (NEAT) is the cumulative energy expended through all other activities of daily living. It is highly variable among individuals; it is controlled by the environment and, possibly, neurobiologically. Mounting evidence suggests that NEAT is critical in determining a person's susceptibility to body fat deposition and is a major factor in human obesity. Recent research supported by the Minnesota Partnership for Biotechnology and Medical Genomics, a state-sponsored collaborative effort of the University of Minnesota and Mayo Clinic, is helping to define the complex brain regulation of NEAT and its role in obesity. This article reviews the evidence for NEAT and the impact of NEAT on obesity. PMID- 16475415 TI - The brain and the biology of obesity. AB - Obesity has emerged as the dominant American public health problem of the 21st century. Because it stems from consuming more energy than is burned, understanding the role of appetite is central to understanding the causes of obesity. Appetite is fundamentally a biological process, based on a set of neurological signals sent from various organs and tissues to the brain; but it is also influenced by environmental and psychosocial factors. This paper outlines the biology of appetite, focusing on the multitude of hormonal and neural pathways that regulate food intake and lead to accumulation of excess body fat. It also briefly explores the effect of psychosocial and environmental factors on the brain and its signaling system, and how researchers are using this information to help control obesity. PMID- 16475417 TI - A pseudo market. PMID- 16475416 TI - Obesity in children: epidemic and opportunity. AB - Obesity has become widespread among both adults and children in the United States, with the prevalence of overweight and obese children increasing from 15% in 1971 to more than 30% in 2000. This article discusses how to determine whether a child is overweight or obese, the causes of childhood obesity, as well as how physicians can treat this disease using family-based diet, exercise, and behavior modification programs; weight-loss medications; protein-sparing modified fasting; and bariatric surgery. PMID- 16475418 TI - Foot and ankle soft-tissue deficiencies: who needs a flap? AB - The patient who needs a flap for a deficit of the foot or ankle soft tissues is any patient who has a break in the integument. While skin grafts often will suffice, if there is a full-thickness loss, particularly over the plantar bony prominence such as the heel or metatarsal, one should strongly consider flaps. In my practice, the flap procedure most commonly performed for the metatarsal region is the V-Y advancement; for the hindfoot, free-tissue transfer; and for the Achilles region, a sural flap. If a lesion is small and the vascular inflow is good, I use free-tissue transfer of a thin skin flap such as a lateral arm flap or a radial forearm flap. For the dorsum of the foot, particularly when there is osteomyelitis or a lesion, or a lateral sidewall lesion, I use a muscle flap. The calcaneus is best served by one of many muscle flaps such as the gracilis. PMID- 16475419 TI - Extended osteotomy for periprosthetic femoral fractures in total hip arthroplasty. AB - Total hip arthroplasty periprosthetic fractures that involve a loose prosthesis and are located at or beyond the tip of the prosthesis are difficult to manage and have higher complication and nonunion rates. In this case report, I describe a fracture-site exposure that allows simple insertion of a long, distally fixed revision prosthesis. In an extended femoral osteotomy, the entire proximal fragment opens the upper segment in "clamshell" fashion. The proximal segment is repaired with or without femoral strut allografts and multiple cerclage wires. With this method, 7 periprosthetic fractures united successfully, and none has required further reconstruction. PMID- 16475420 TI - Multimodal venous thromboembolic disease prevention for patients undergoing primary or revision total joint arthroplasty: the role of aspirin. AB - Venous thromboembolic disease (VTD), which consists primarily of deep venous thrombosis (DVT) and pulmonary embolism (PE), is of significant concern to orthopedic surgeons who perform total hip and total knee arthroplasties. DVT and PE can be prevented in multiple ways; each method or combination of methods has its benefits and drawbacks. Seemingly, the more efficacious a medication or method for preventing VTD, the higher the associated risk for adverse events such as bleeding and wound complications. For each patient, then, the balance or homeostasis between significant clotting event and bleeding must be determined. Examining this balance and understanding the benefits and risks associated with each medication or intervention may allow surgeons to make educated decisions about prophylaxis for their patients. Furthermore, risk stratification and multimodal management may prove to be the safest and most effective way to manage VTD prevention. PMID- 16475421 TI - Vascular risk associated with bicortical tibial drilling during anteromedial tibial tubercle transfer. AB - For select patients with persistent patellofemoral pain, the anteromedial tibial tubercle transfer (Fulkerson osteotomy) provides excellent clinical results. This procedure, indicated for patients with patellar malalignment, has become one of the most popular distal realignment procedures. One potential concern with this technique is the proximity of the posterior vascular structures during bicortical tibial drilling for screw placement. To address this concern, we measured the proximity of these structures in 7 consecutive fresh-frozen cadaveric knees. For each knee, barium was injected into the femoral artery, and anteroposterior (AP) radiographs were taken to document the location of the popliteal vessels. Next, the initial steps of the Fulkerson osteotomy were performed. Then, a lateral release and the tibial osteotomy were performed, the tubercle was advanced into position, and two 9/64-inch extralong drill bits were placed through the tubercle and the posterior tibial cortex. Repeat AP radiographs were obtained, and digital calipers were used to measure the distance from the drill bits to the popliteal vessels. The vascular structure closest to the exit point of the superior drill bit was the bifurcation of the popliteal artery (mean distance, 8.3 mm; SD, 9.3 mm; range, 0.0-21.3 mm), and in 2 knees this structure directly overlay the bifurcation on the AP radiograph; the vascular structure closest to the exit point of the inferior drill bit was the posterior tibial artery (mean distance, 9.0 mm; SD, 8.0 mm; range, 0.0-20.0 mm), and again in 2 knees the drill bit lay directly over the artery on the AP radiograph. Bicortical drilling for screw placement during the anteromedial tibial tubercle transfer procedure may come precariously close to the posterior vascular structures of the knee, so orthopedic surgeons must take extreme caution not to drill past the posterior cortex during this part of the operation. PMID- 16475422 TI - Posterior circumflex humeral artery aneurysm in 2 professional volleyball players. PMID- 16475424 TI - Guide-wire exchange before insertion of intramedullary nails. AB - In the past, before a reamed intramedullary nail was inserted, it was mandatory to exchange the beaded-tip reaming guide wire with a smooth-tip wire. This additional step is optional in most new nail designs. Here we describe 2 cases in which not exchanging the wire led to initial wire incarceration and introduced additional steps that had to be taken before locked nailing could be completed. Intramedullary nail designs that avoid extra steps and thereby promote simplicity and reliability of insertion are potentially advantageous to both patient and surgeon. Eliminating the exchange tube and the smooth wire, however, should be approached with caution, as no time may be saved, and the procedure may be made much more difficult technically. PMID- 16475423 TI - Using flexible nails to improve stabilization of proximal femur fractures in adolescents. AB - Intramedullary fixation as a treatment for adolescent femur fractures has increased in popularity. We investigated the varus stability of 3 fixation techniques--the standard 2-nail configuration (2 retrograde nails), a 3-nail configuration (standard 2-nail configuration plus 1 antegrade nail), and a retrograde/antegrade 2-nail configuration--in synthetic femurs with 2 simulated fracture patterns (transverse, butterfly). A varus moment was applied to the specimens until 20 degrees of varus angulation was obtained. The varus angular stiffness of the construct defined varus stability. The 3-nail configuration had the most varus angular stiffness with both fracture patterns. The varus stability of the 3-nail configuration was statistically higher than that of the standard 2 nail configuration with the transverse fracture pattern only. Adding 1 antegrade nail to 2 retrograde nails significantly increased resistance to varus deformity of reduced proximal-third transverse femur fractures in the synthetic femur model. PMID- 16475425 TI - Nurse prescribing is in dispute, but why, exactly? PMID- 16475426 TI - Competition--the 'missing link' in the NHS. PMID- 16475427 TI - Patients' satisfaction with a nurse-led oncology service. AB - This study reports the findings of a quantitative study determining the satisfaction levels of patients attending a nurse-led oncology day ward. A random sample of 100 patients was surveyed using an adapted version of the Leeds Satisfaction Questionnaire (Hill, 1997) to ascertain the satisfaction levels of patients attending the unit. Satisfaction levels were found to be favorable in general. However, the study did highlight that some aspects of patient information required attention. Moreover, issues with the use of the questionnaire emerged as the inclusion of both positive and negative statements in the Likert-style questionnaire presented contradictory findings. PMID- 16475428 TI - Purposful dehydration in a terminally ill cancer patient. AB - The management of terminally ill patients who are unable to maintain their own hydration, or who express a wish to discontinue artificial hydration, is a significant controversial topic among healthcare professionals, patients and families. Evidence-based research suggests a number of medical and nursing indications and contraindications for dehydration. The purpose of this article is to review the current research, consider the factors involved in decision-making and clarify the complex issues which have implications for nursing surrounding the management of dehydration in terminally ill patients. The article uses the case of a patient nursed by the author and is written as a reflective account of practice. PMID- 16475429 TI - Clinical corner: medication-induced hyponatraemia. PMID- 16475430 TI - What happens to used colostomy pouches outside the acute setting. AB - This article looks at the disposal of used colostomy pouches once a patient is discharged from the acute setting. An overview of the guidance given by stoma care nurses to colostomy patients prior to discharge is given as part of a combined study by a patient association. There are approximately 100,000 ostomates in the UK, although the exact number is not known (McCahon, 1999). Approximately 50% of these are colostomates. There appears to be very little literature on disposability of used stoma pouches or advice given to the ostomate with a newly formed colostomy. PMID- 16475431 TI - Genetics knowledge for nurses: necessity or luxury? AB - Senior nurses within frontline NHS Trusts in East Anglia were consulted about their perceptions of the genetics-related knowledge and education required by nurses working in their Trusts. They disagreed with policy decision makers, i.e. the Government and genetics specialists, on the currently necessary knowledge levels. They also did not recognize genetics as an important issue in current everyday nursing activities. The difference in perception also related to the speed with which its presence and importance will increase. Nurses already working under considerable pressure while not perceiving the importance of genetics in everyday practice cannot be expected to willingly absorb new genetics related knowledge. The National Genetics Education and Development Centre in Birmingham will have to reach out to practitioners and managers throughout the UK and establish a shared platform from which to develop this issue in the future. PMID- 16475432 TI - Mental health records. AB - Records relating to the detention and treatment of mentally disordered persons are one of the few occasions on which there are statutory provisions over what must be recorded and in what circumstances, and how corrections can be made. In the event of a failure to comply with the statutory provisions relating to detention, the patient would not be lawfully detained and could seek his/her release. Where a patient is admitted informally to a psychiatric hospital, there are no statutory provisions but the general principles discussed in an earlier article relating to record keeping should be observed. PMID- 16475434 TI - Practice nurse who expanded her role without appropriate training. PMID- 16475433 TI - Using the 12-lead ECG to assess acute coronary patients. AB - Acute coronary syndromes (ACS) are characterized by the rupture of unstable plaque within coronary arteries. Depending on the extent of the ensuing occlusion and myocardial damage, ACS can be classified as unstable angina, non-ST elevation myocardial infarction (MI) and ST elevation MI. The electrocardiogram (ECG) is an invaluable tool in the assessment of patients with ACS. It provides evidence for the location of myocardial ischaemia, injury and infarction and is a crucial factor in the decision to administer thrombolytic agents and other management strategies. The 12-lead ECG is limited to a view of the left ventricle, however it can be extended to provide additional information about the right ventricular and posterior walls. Critical care nurses with ECG skills can contribute to the early detection and management of patients with ACS. PMID- 16475435 TI - Combination model of care for community nurse practitioners. AB - Twenty-two community nurse practitioners were researched, primarily to establish whether or not they were able to achieve a 'higher level of practice'. Grounded theory, the qualitative research methodology developed by Glaser and Strauss, was used as a framework for the research. Data was gathered using primary observation with some participation (581 consultations being observed in the process), and by interviewing the community nurse practitioners and their clinical managers. From the overall theoretical framework that emerged from the research, a category, the 'combination model of care', was developed as a theoretical model, together with a conceptual framework for the process of nursing, 'HADPIPE'. This model of care/nursing and the HADPIPE framework provide a basis from which nurse practitioners can practice with a holistic nursing focus. The article analyses and discusses the combination model, and provides a detailed example of it in action. PMID- 16475436 TI - The safety and dignity of patients and nurses during patient handling. AB - Since the implementation of the Moving and Handling Operations Regulations (Health and Safety Executive, 1992) there has been considerable effort to reduce back injury among nurses by developing patient handling policy that eliminates actual manual handling. Recently, however, there have been challenges to 'no manual handling' policies by patients as an issue of dignity and human rights. This article offers a literature-based discussion of patient handling in relation to safety and dignity for both patients and nurses. The literature does not offer a solid evidence base for handling practice, and research, involving patents, is limited. However, nurses' handling practice is governed by legislation and NHS Trust policies. Patient handling that is safe and dignified for both patient and handler is a vital skill that requires knowledge and competence. This article suggests ways of developing practice that is safe and dignified for both patient and handler. PMID- 16475437 TI - NMC consultation on fitness to practice. PMID- 16475438 TI - A new document for the New Year. PMID- 16475439 TI - Personalized drug therapy with pharmacogenetics. Part 1: Pharmacokinetics. PMID- 16475440 TI - The healing power of dogs: Cocoa's story. AB - Animals bring a sense of "at homeness" and even normalcy to people who are hospitalized or live in health care facilities. Illnesses, separation from family, fear, loneliness, and even depression may be lessened for those who receive a therapy dog visit by providing a welcome change in routine and something to look forward to. Individuals are often more active and responsive during and after a visit. Just stroking and petting a dog requires the use of hands and arms, as well as the motions of stretching and turning. Dogs are also unconcerned with age or physical ability; they accept people as they are. This alone causes many to reach out and interact with dogs. Animals provide a focus for conversation and a common interest. As Cocoa's human partner can attest, a pet also makes it easier for strangers to talk. Cocoa is known by so many people at the facilities she visits that residents' family members often stop to talk with "Cocoa's mom" when they meet her in the community, asking questions about how Cocoa is doing. When Cocoa is no longer able to perform her services, many in her community will mourn. Cocoa's story, as well as the limited research literature, supports the positive physical and psychological effects of animals, but continued research on the effect of therapy animals on health outcomes is needed. As advocates for clients, nurses are in key positions to facilitate the inclusion of animals in clients' care. Although not for everyone, including those who react negatively to animals or who are allergic, pet therapy offers important possibilities for providing holistic care that extends not only to clients, but also to family members and staff, and to the pets themselves. PMID- 16475441 TI - Hyper-anxiety in early sobriety: psychopharmacological treatment strategies. AB - The importance of recognizing and treating anxiety disorders that co-occur with alcohol use disorders should not be underestimated, whether the anxiety is state (a transient condition) or trait (a chronic disorder). This article will discuss hyper-anxiety, a term the authors use to describe the uncomfortable, excessively anxious state that contributes to relapse. In addition, psychopharmacological strategies for relief of hyper-anxiety are suggested, and implications for recovery are discussed. PMID- 16475442 TI - Moving toward recovery within clients' personal narratives: directions for a recovery-focused therapy. AB - Recent literature emphasizes that recovery from schizophrenia involves recovery within one's own narrative of an integral sense of identity, agency, social connection, and worth. While this is intuitively appealing and consistent with a wide range of literature, it raises the issue of how to best help people do this in individual psychotherapy. In this article, we explore how psychotherapy might help people construct new narratives or storied understandings of their lives and thereby promote recovery from schizophrenia. Exemplified with two individual examples, we first discuss the barriers that challenge and the techniques that help psychotherapists seeking to enter into dialogue with people with severe mental illness. We also offer a theoretical model of how the revitalization of dialogues within therapy can be conceptualized as a process that promotes recovery and discuss the objective measurement of such outcomes. PMID- 16475443 TI - Telecare for women with postpartum depression. AB - Data were collected to pilot test the feasibility and effects of telecare as an intervention for depression in a small group of mothers with postpartum depression. Treatment involved a 10-week telecare therapy consisting of three related aspects: cognitive-behavioral therapy, relaxation techniques, and problem solving strategies. Beck Depression Inventory II scores were significantly lower after telecare treatment. Women identified psycho-education as the greatest help to them. PMID- 16475445 TI - Health plans, CMS agree to reimburse States' stopgap drug coverage for dual eligibles. PMID- 16475444 TI - Hoarding as an adult: overview and implications for practice. PMID- 16475446 TI - Prescription drug package inserts get a user-friendly makeover. PMID- 16475447 TI - E-prescribing goes under the microscope. PMID- 16475448 TI - Cognitive impairment is associated with poor oral health in individuals in long term care. AB - PURPOSE: The aim of this work was to study oral hygiene and cognition in patients in long-term care. MATERIALS AND METHODS: A representative sample of individuals in long-term care aged 67 years and above, from all 19 countries of Norway, was selected for the study. Trained local dental teams examined 1910 individuals- 1358 from institutions and 552 living at home. Oral hygiene was assessed by means of a combined plaque and mucosal score (MPS), and cognition by means of a short version of Mini-Mental State Examination (MMSE-12). RESULTS: Mean age was 84.2 years and the age range was 67-106 years. Over 70% were cognitively impaired. Cognitive impairment increased with age and was more prevalent in institutionalised individuals compared to those living at home. Poor oral status was more prevalent among individuals with cognitive impairment. CONCLUSION: The results of this study indicate that a large proportion of individuals in long term care in Norway show cognitive impairment. A high proportion of these has a poor oral status. Thus, they carry a high risk of developing dental caries. Obviously, there is a demand for careful and close surveillance of oral hygiene and oral health in individuals in long-term care, and especially of those cognitively impaired. PMID- 16475449 TI - Self-efficacy perspective on oral health among Turkish pre-adolescents. AB - OBJECTIVE: To assess the association between toothbrushing and dietary self efficacy and oral health status and behaviour related to it among Turkish pre adolescents. MATERIALS AND METHODS: A cross-sectional study of Turkish school pre adolescents aged 10 to 12 (n = 611) by oral health examinations (OHE) and self administered health behaviour questionnaire (PHBQ). The PHBQ, including new Toothbrushing (TBSES), and modified Dietary Self-Efficacy (DSES), was designed to survey pre-adolescents' oral, dietary and general hygiene behaviour by psychosocial factors. An OHE based on World Health Organization criteria was implemented. The participation rate was 97% (n = 591), and 95% (n = 584) for PHBQ and OHE. Multiple linear regression analysis, descriptive statistics, factor and item analysis, Pearson correlation coefficient, chi-square test, and Student t test were applied. RESULTS: Pre-adolescents, mostly with regular toothbrushing habits (69% at least once a day) and high levels of irregular dental visits (79%) had as their mean DMFS figure 3.77 +/- 3.37. Principal component analysis for TBSES and DSES revealed that four factors met the Kaiser Criteria, accounting for 50.2% of total variance. Scales with good internal consistency and test-retest stability were associated with professionally (DMFS) and self-assessed dental health (p < 0.05). A similar tendency appeared between self-assessed gingival health and DSES (p < 0.05). CONCLUSION: Self-efficacy beliefs were associated with oral health and related behaviour among pre-adolescents. Further research is needed for implementation of scales to enhance oral health. PMID- 16475450 TI - Prevalence of malocclusions and their impact on the quality of life of 18-year old young male adults of Florianopolis, Brazil. AB - PURPOSE: To evaluate the prevalence and the impact of malocclusions on the quality of life of 18-year-old young male adults. MATERIALS AND METHODS: Cross sectional study involving young adults who applied for military service in the city of Florianopolis in the year 2003 (N = 3735). The sample size was 414 individuals randomly selected during the health examination days. The malocclusion data were obtained through the Dental Aesthetic Index variables. The impact of the oral health conditions on the quality of life was obtained through the Oral Impact on Daily Performance. RESULTS: The highest prevalence was the molar relationship in mesio- or disto-occlusion with 57.3% (95% CI 52.5 - 62.1); 36.4% (95% CI 28.6% - 4.2%) of the applicants reported an impact on their quality of life as a result of the oral health condition. Results from the multiple logistic regression analysis showed that only the incisal crowding [OR 1.9 (95% CI 1.2 - 3.1)] (p = 0.008) showed an impact on the applicants' quality of life. CONCLUSION: The malocclusion of highest prevalence was molar relationship disorders (57.3%), and only the dental crowding generated an impact on the quality of life. PMID- 16475451 TI - Developments in oral health policy in the Nordic countries since 1990. AB - PURPOSE: There is a number of systems for the provision of oral health care, one of which is the Nordic model of centrally planned oral health care provision. This model has historically been firmly based on the concept of a welfare state in which there is universal entitlement to services and mutual responsibility and agreement to financing them. This study reports and analyses oral health care provision systems and developments in oral health policy in the Nordic countries (Denmark, Finland, Iceland, Norway and Sweden) since 1990. MATERIAL AND METHODS: Descriptions of and data on the oral health care provision systems in the Nordic countries were obtained from the Chief Dental Officers of the five countries, and contemporary scientific literature was appraised using cross-case analyses to identify generalisable features. RESULTS: It was found that in many respects the system in Iceland did not follow the 'Nordic' pattern. In the other four countries, tax-financed public dental services employing salaried dentists were complemented by publicly subsidised private services. Additional, totally private services were also available to a variable extent. Recently, the availabilty of publicly subsidised oral health care has been extended to cover wider groups of the total population in Finland and Sweden and, to a smaller extent, in Denmark. Concepts from market-driven care models have been introduced. In all five countries, relative to the national populations and other parts of the world, there were high numbers of dentists, dental hygienists and technicians. Access to oral health care services was good and utilisation rates generally high. In spite of anticipated problems with increasing health care costs, more public funds have recently been invested in oral health care in three of the five countries. CONCLUSION: The essential principles of the Nordic model for the delivery of community services, including oral health care, i.e. universal availability, high quality, finance through taxation and public provision, were still adhered to in spite of attempts at privatisation during the 1990 s. It appeared that, in general, the populations of the Nordic countries still believed that there was a need for health and oral health care to be paid for from public funds. PMID- 16475452 TI - The effect of different methods of drinking a carbonated beverage on the pH of dental plaque: an in vivo study. AB - PURPOSE: Fizzy drinks are known to be erosive or cariogenic, but little is known about the ways of reducing their harmfulness by altering the method of drinking. The purpose of this study was to assess the changes in plaque pH, at different time intervals in vivo after consuming a carbonated beverage (sprite, pH = 2.98) with plastic glass, straw and directly from bottle. DESIGN: A clinical study. MATERIAL AND METHODS: Eighteen subjects aged 18-25 years were recruited for the study and were divided randomly into three groups, six in each (group A- plastic glass, B- straw and C- directly from bottle) after the salivary pH was measured. Subjects were requested to refrain from brushing for 24 hours prior to the study. Collection of pooled plaque was done before and after consuming the drink at five, 10-, 20- and 30-minute intervals. Plaque pH was assessed by glass combination electrode. ANOVA and post hoc Tukey's test was used for statistical analysis. RESULTS: Highest mean pH drop (5.29) was recorded when consumed with plastic glass at all time intervals. There was a significant difference between group A and B at 5 min and 10 min (P < 0.05). However, no difference was seen between group B and C, A and C (P > 0.05). CONCLUSIONS: The use of a straw and direct consumption of beverage from the bottle could limit harmful effects on dentition. PMID- 16475453 TI - Dental caries and orofacial pain trends in 12-year-old school children between 1997 and 2003. AB - PURPOSE: To estimate the prevalence of dental caries and orofacial pain in 12 year-old schoolchildren in the Southern Brazilian town of Palhoca in 2003 and to compare it with results from 1997. MATERIALS AND METHODS: A cross-sectional study was carried out involving 444 randomly selected 12-year-old schoolchildren in 2003. WHO (1997) criteria for dental caries and Locker and Grushka (1987) criteria for orofacial pain were used. Chi-square and Fisher exact tests were used to examine association between variables. To test the independence of variables, logistic regression analysis was performed. RESULTS: The prevalence of caries was 55.8%. The mean DMF-T was 1.65. Component D was the one that most contributed to the indicator. In 1997, caries prevalence was 73.5% and the mean DMF-T was 2.84. Similarly, the component that most contributed to the indicator was D with 74.0%. In the trend analysis a reduction of 24.1% in the caries prevalence and 41.9% in the severity measured by the DMF-T was observed. The prevalence of orofacial pain was 66.6%. In 1997, the prevalence was 79.5%. In both years the most common type of pain was stimulated toothache. A reduction of 12.9% in the prevalence of orofacial pain and 28.3% in stimulated toothache was observed. Results of the logistic regression analysis showed that orofacial pain was associated with dental caries independently of other studied variables. CONCLUSION: A reduction in both caries and orofacial pain in the studied period was observed. Orofacial pain was independently associated with the presence of caries. PMID- 16475454 TI - Caries status of children in eastern Trinidad, West Indies. AB - PURPOSE: To determine the prevalence, severity and caries burden among children residing in eastern Trinidad. MATERIALS AND METHODS: This is a cross-sectional study comprising probability, stratified, and proportionate-to-size sampling. Seven hundred and eleven school children aged six, 12 and 15 years were examined between November 1999 and January 2000 by two calibrated dentists using World Health Organization (WHO) assessment criteria. RESULTS: Overall mean DMFT and dft scores were 1.29 +/- 2.3 and 1.78 +/- 3.1 while 30.4% were caries-free. At ages six, 12 and 15 years, mean DMFT scores were 0.08 +/- 0.38; 2.18 +/- 2.49; 2.66 +/ 3.0 while dft scores were 3.74 +/- 3.63; 0.14 +/- 2.65; 0. Significant Caries Indices (SiC) for permanent and primary teeth were 3.75 and 5.28. Children of parents/carers with university education had the lowest DMFT/dft scores (1.0 +/- 2.47/0.83 +/- 1.58), treatment need (23.8%), and comprised the highest proportion of caries-free (76.2%) population. The values for father/male carer were statistically significant (p < 0.005). All four first permanent molars were decayed in 8.76%. First permanent molars were most frequently decayed in 12-year olds (55.4%) and 15-year-olds (50.0%). Untreated caries and first permanent molar decay among six-year-olds were 84% and 8.3% respectively. Tooth mortality was 0.978 and 0.964 in permanent and primary teeth. The Restorative Indices for permanent and primary teeth were 0.02134 and 0.0213. CONCLUSION: Based on WHO severity criteria for the respective ages, caries experience is low in Eastern Trinidad. The mean DMFT is much lower than recently reported country data for 12 year-olds. However, the incidence is increasing. The SiC suggests that a small proportion of the population accounts for most of the decayed teeth. PMID- 16475455 TI - Effect of a stannous fluoride dentifrice on the sulcular microbiota: a prospective cohort study in subjects with various levels of periodontal inflammation. AB - OBJECTIVES: To assess the effects of an experimental 0.454% stannous fluoride (SnF2) dentifrice on the oral sulcular microbiota in patients with various stages of oral diseases using checkerboard DNA-DNA hybridization. MATERIAL AND METHODS: In the present one-month, single center, single product, prospective cohort trial, 37 adults (mean age 37.6) were assigned to one of four oral health condition cohorts with seven to 10 subjects each: 1. mild gingivitis, 2. marked generalized gingivitis to moderate periodontitis, 3. caries-prone and 4. treated moderate to advanced chronic periodontitis in supportive periodontal care. All four groups were asked to use the test dentifrice and a power toothbrush twice a day for one minute during a four-week test period. Before and after the trial period, Plaque Indices (PII, Silness and Loe, 1964) and Gingival Indices (GI, Loe and Silness, 1963) were recorded. Subgingival plaque samples were collected from all patients at Baseline, as well as after two and four weeks. These samples were analyzed for content of 40 bacterial species using checkerboard DNA-DNA hybridization. RESULTS: As a result of the only one minute brushing with the stannous fluoride dentifrice, the mean PII at Baseline was significantly lower (p < 0.05) from the mean PII at four weeks. No statistically significant differences were found between premolar and molar mean values. Moreover, no statistically significant differences were found between the mean GI at Baseline and at four weeks. The microbiological analysis showed that at baseline subjects in groups 2 and 4 had significantly higher bacterial loads of bacteria than groups 1, and 3 (i.e. A. actinomyctemcomitans P. gingivalis, T. forsythia, and T. denticola. Over the study period, the total bacterial load did not change in groups 2, 3 and 4. In groups 1 and 3, however, an increase in the loads of Streptococci spp. were noticed (p < 0.05) including S. mitis, S. intermedius, and S. sanguis (p < 0.01) suggesting an increase in the presence of early colonizing and health associated bacteria. CONCLUSION: One minute brushing with a 0.454% stannous fluoride dentifrice did--after four weeks--not affect the subgingival microbial profiles in patients with moderate periodontitis and treated moderate to advanced periodontitis. However, the sulcular microbial profiles of mild gingivitis and caries-prone patients were affected, indicating a shift towards a gingival health associated microbiota in the sulcular region of patients not affected by attachment loss. RUNNING HEAD: Effect of stannous fluoride on sulcular microbiota. PMID- 16475456 TI - Immediate antecedents: just part of the picture. PMID- 16475457 TI - A limited look at falls. PMID- 16475459 TI - Legal issues in gerontological nursing. Part 1: Abuse and neglect of older adults. PMID- 16475460 TI - A typology of oral hydration problems exhibited by frail nursing home residents. AB - Dehydration remains a substantial problem for nursing home residents, often with poor health outcomes. The purpose of this investigation was to establish 6-month prevalence of dehydration events in nursing home residents and to describe common hydration problems of nursing home residents. In this prospective observational study, 35 nursing home residents were followed for 6 months to assess problems with hydration and to evaluate the presence of dehydration. Urine specific gravity and color, bioimpedance measurements, meal intake recordings, and chart abstraction were used to assess hydration status. Field notes and informal staff interviews were used to describe specific hydration problems and clinically relevant interventions. Dehydration events occurred in 31% (11 of 35) of residents during the 6-month period. A typology of hydration problems was developed from the field observations. The typology consists of four groups (i.e., Can Drink, Can't Drink, Won't Drink, End of Life) and six corresponding subgroups. Demographic and hydration characteristics of the subgroups were compared and contrasted. Comparisons revealed the Won't Drink group is most vulnerable to dehydration events because this group has the highest percentage of dehydration events (58%, 4 of 7), the highest average specific gravity, and the lowest consumption of fluids during meals. Nursing interventions for the subgroups are discussed. Targeting nursing interventions to the specific hydration problem exhibited is proposed. PMID- 16475461 TI - Undernutrition in older adults across the continuum of care: nutritional assessment, barriers, and interventions. AB - Undernutrition can be a significant deterrent to healthy aging and can nega tively affect health outcomes in older adults. Researchers have identified the prevalence of undernutrition in older adults and the need for intervention yet the incidence remains high. The purpose of this article is twofold: to emphasize the need for nutritional assessment across the continuum of care a experienced by older adults, and to identify possible barriers to effective treatment. The assessment of nutritional status and the implementation of effective nutritional interventions are essential to the health of older adults. PMID- 16475462 TI - Bladder control educational needs of older adults. AB - Bladder control problems, especially urinary incontinence (UI) and urinary freqency and urgency, are prevalent among older adults. The purpose of this project was to determine older adults' educational needs, preferred format, and interest in bladder control educational materials. Eighty-one older adults participated in focus groups conducted at senior centers, churches, and senior apartment dwellings located in Philadelphia, PA. A doctorally prepared nurse acted as the moderator, using a questionnaire that included items on general health issues and specific questions about bladder control issues. Participants showed interest in learning more about bladder control issues, discussed their preferences in relation to format for receiving health-related information, and expressed their interest in bladder control educational materials. This information can be used to devise an educational program about bladder control that is acceptable to older adults and can be easily accessed in settings where they reside or recreate. PMID- 16475463 TI - Overcoming restraint use: examining barriers in Australian aged care facilities. AB - The purpose of this article is to discuss the outcomes of a project that sought to identify alternatives to restraint use in aged care facilities. (In Australia, the term aged care facility refers to those facilities previously known as hostels and nursing homes.) A literature review was conducted and discussion forums and individual interviews were held with interested stakeholders in Australian capital cities, as well as in a rural city in each state. Site visits were conducted at 16 aged care facilities. Participants identified legal concerns, existing organizational culture, and lack of alternatives as barriers to implementing restraint-minimization practices. Recommendations to facilitate minimal restraint are implementing national policy guidelines, adopting a best practice philosophy in relation to restraint minimization, continuing staff education related to alternatives, and maintaining constant communication with families. PMID- 16475464 TI - Conflict resolution styles: a comparison of assisted living and nursing home facilities. AB - In this exploratory study, the authors investigated how interpersonal conflict is resolved in assisted living and nursing home facilities. In particular, the authors examined whether conflict resolution styles differed between type of facility and between residents and staff in each type of facility. Four focus groups were conducted--two with residents and two with staff from each type of facility. The focus groups centered on discussing the occurrence of conflict and how each participant handled it. Discourse analysis was employed to identify participants' use of three styles of conflict resolution: controlling, solution oriented, and non-confrontational. The results indicate that staff in each care context showed a preference for the solution-oriented approach. Residents in each setting reported equal use of the non-confrontational and solution-oriented styles. The findings suggest that preferred conflict resolution styles may vary more as a function of the role of each communicator than the context of the care setting. PMID- 16475465 TI - The experience of being listened to: a qualitative study of older adults in long term care settings. AB - The experience of being listened to for older adults living in long-term care facilities was explored using a qualitative descriptive method outlined in Parse (2001), with the human becoming theory as the theoretical framework. The themes that emerged from this study--Nurturing Contentment, Vital Genuine Connections, and Deference Triumphs Mediocrity--affirmed the experience of being listened to as fundamental to the participants' quality of life. The findings expand nursing theory, provide enhanced understanding of the experience of being listened to, and offer ideas for future research. Through the voices of older adults participating in this study, the authors learn how critical listening is to quality care, and thus to excellence in nursing practice. PMID- 16475466 TI - If it's Thursday, I must be at the senior center. PMID- 16475467 TI - Significance of combined angiotensin II receptor blocker and carvedilol therapy in patients with congestive heart failure and arginine variant. AB - OBJECTIVE: Angiotensin II receptor blockers (ARBs) and beta-blockers have contributed to longer life expectancies for patients with congestive heart failure. However, whether the use of ARBs is helpful for introducing carvedilol (beta-blocker) is unclear when patients with symptomatic congestive heart failure are admitted to the hospital. METHODS: In this retrospective study, 27 patients with symptomatic congestive heart failure were given carvedilol upon admission. Five patients received carvedilol monotherapy (group A), and 22 were treated with a combination of carvedilol and ARBs (group B). RESULTS: There was no difference in medication between the groups except for ARBs. In addition, there were no significant differences in the decrease in plasma brain natriuretic peptide, or the improvement of left ventricular ejection fraction upon carvedilol treatment between the groups. Although there was no significant difference in the maintenance dose of carvedilol between the groups, the gross dose of carvedilol in group B was significantly lower than that in group A. In addition, the improvement of left ventricular ejection fraction in group B was positively correlated with the maintenance dose of carvedilol in patients who had wild-type beta1-adrenergic receptor at amino acid 389 (arginine/arginine genotype). CONCLUSIONS: These results suggest that ARBs are helpful for introducing carvedilol in patients with the wild-type beta1-adrenergic receptor gene, and that treatment with combined treatment with ARB or analysis of the beta1 adrenergic receptor genotype may offer advantages to control congestive heart failure in the short-term. PMID- 16475469 TI - [Follow-up study of diastolic paradoxical flow in the left ventricle with apical hypertrophic cardiomyopathy]. AB - OBJECTIVES: Left ventricular apical asynergy and cardiovascular complications were evaluated in patients with apical hypertrophic cardiomyopathy, in whom apical diastolic paradoxical flow toward the base could be detected by Doppler color flow echocardiography. METHODS: Twenty-nine patients with apical hypertrophic cardiomyopathy were followed up with echocardiographic examinations for at least 5 years. They were divided into three groups: those who persistently exhibited paradoxical flow (Group A, n = 13), those in whom paradoxical flow developed during the follow-up period (Group B, n = 8), and those in whom paradoxical flow was not detected during the follow-up period (Group C, n = 8). Peak flow velocity and duration of paradoxical flow, the presence or absence of apical asynergy, and cardiovascular complications were evaluated. RESULTS: The mean follow-up period was 7.3 +/- 1.4 years. In Groups A and B, the apical wall motion deteriorated over time, whereas there was no asynergy change in Group C. Further, peak flow velocity and duration of paradoxical flow increased throughout the follow-up period in Groups A and B, and were correlated with the severity of apical asynergy. Ventricular tachycardia and cerebrovascular complications also occurred more often in patients with paradoxical flow. CONCLUSIONS: The presence of paradoxical flow was related to the severity of apical asynergy and cardiovascular complications, and may be an important marker for evaluating the clinical course of patients with apical hypertrophic cardiomyopathy. PMID- 16475468 TI - [Dilation of the brachial artery in response to sublingual nitroglycerin can predict the antihypertensive effects of valsartan: a study using novel high frequency high-frame-rate ultrasound imaging]. AB - OBJECTIVES: The efficacy of antihypertensive agents can vary in patients. Four to 8 weeks may be required before antihypertensive agents become fully effective. Predicting the efficacy can help agent selection and dose setting. This study determined whether nitroglycerin-induced vasodilation of brachial arteries can predict the antihypertensive action of angiotensin II receptor antagonist. METHODS: Untreated uncomplicated patients with essential hypertension, who gave informed consent, were studied (n = 20, mean age 55 years). Before antihypertensive treatment, nitroglycerin-induced vasodilation of the brachial arteries was measured using a novel method of 15 MHz high-frequency high-frame rate ultrasound imaging (Hitachi EUB8000). Diameter of the brachial artery at the end-systolic phase was measured before and after 0.3 mg nitroglycerin sublingual spray and percentage vasodilation (%D-N) was calculated. The reduction of mean blood pressure after nitroglycerin (%BP-N) was calculated. Valsartan monotherapy (40-80 mg/day)was administered for 3-6 months (mean 132 days). Reduction of mean blood pressure after valsartan monotherapy (%BP-V) was calculated. RESULTS: Valsartan decreased systolic blood pressure from 138 +/- 13 to 130 +/- 17 mmHg, and diastolic blood pressure from 83 +/- 11 to 78 +/- 11 mmHg (p < 0.05). %D-N was correlated closely with %BP-V (r = - 0.70, p < 0.001). %BP-N had no correlation with %BP-V (r = 0.13, p = 0.58). CONCLUSIONS: Direct vasodilatory action of nitroglycerin on vascular smooth muscle cells may predict the chronic antihypertensive effect of angiotensin II receptor antagonist. PMID- 16475470 TI - [Improvement of central sleep disordered breathing with severe congestive heart failure by biventricular pacing therapy: a case report]. AB - A 74-year-old man with ischemic cardiomyopathy was repeatedly admitted for congestive heart failure. His left ventricular ejection fraction was 21% and diastolic left ventricular dimension was 73.5mm by echocardiography. He was treated with biventricular pacing and heart failure improved from New York Heart Association class III to II. Before the treatment, brain natriuretic peptide was 600.5 pg/ml. Apnea hypopnea index was 23.8 and all events were central type. After biventricular pacing, apnea hypopnea index was improved to 21.9 after 11 days, 14.0 after 33 days, and 4.8 after 48 days. His left ventricular ejection fraction was 36%, diastolic left ventricular dimension was 71.4mm, and brain natriuretic peptide was 383.8 pg/ml. In this patient, central sleep disordered breathing was improved by biventricular pacing therapy after only 48 days. PMID- 16475471 TI - Recurrent severe mitral regurgitation due to left ventricular apical wall motion abnormality caused by coronary vasospastic angina: a case report. AB - A 76-year-old man developed congestive heart failure due to severe mitral regurgitation after episodes of vasospastic angina. Echocardiography demonstrated left ventricular apical akinesis with ballooning and deformity of the anterior mitral leaflet becoming concave toward the left atrium. The acetylcholine provocation test induced diffuse coronary vasospasm in the distal segments of both right and left coronary arteries and reproduced severe mitral regurgitation. Follow-up echocardiography demonstrated decreased mitral regurgitation with ameliorated apical wall motion. Coronary vasospasm remained refractory to antivasospastic medications and severe mitral regurgitation relapsed 1 month after discharge. Mitral valve annuloplasty with a Carpentier-Edwards physio ring was performed, and no recurrence of mitral regurgitation was observed despite some episodes of vasospastic angina. We speculate that vasospastic angina and the resultant apical wall motion abnormality caused tethering of the mitral subvalvular apparatus, leading to inappropriate mitral coaptation and severe regurgitation. PMID- 16475472 TI - [Takotsubo cardiomyopathy manifesting as no reflow pattern in coronary flow by transthoracic Doppler echocardiography and prolonged recovery of regional left ventricular wall motion abnormality: a case report]. AB - A 80-year-old woman was admitted to our hospital because of chest pain. Electrocardiography revealed ST segment elevation in the I, aVL, and V1-V5 leads. Echocardiography revealed left ventricular apical aneurysmal change with ejection fraction of 31%. Coronary angiography showed no abnormalities. Creatine kinase was not elevated in her clinical course. The diagnosis was takotsubo cardiomyopathy. Transthoracic Doppler echocardiography was performed on the 2nd hospital day. Coronary flow velocity pattern in the left anterior descending artery revealed shortened diastolic deceleration time (108 msec) and systolic retrograde flow. Asynergy of the left ventricle gradually improved, but still persisted slightly at 6 months after discharge. Most patients with takotsubo cardiomyopathy have normal coronary flow velocity pattern in the acute phase. In this case, no reflow pattern of coronary flow was observed during prolonged recovery from left ventricular regional wall motion abnormality. PMID- 16475473 TI - [Unusual echo-free space behind the left atrium]. PMID- 16475474 TI - Cheating: an ethical concern for nursing educators. PMID- 16475475 TI - A report addressing the resources needed to increase the capacity of the Kansas Board of Regents system for educating registered nurses. Charge from the Legislative Budget Committee. PMID- 16475477 TI - Comparison of performance characteristics between first- and third-generation hematology systems. AB - In 1996, Barnes-Jewish Hospital introduced the first fully robotic hematology system in North America. This first-generation Coulter/IDS robotic automated system consisted of a series of transport lanes, inlet and outlet stations, 2 robotic arms, and an on-line slide maker/strainer. The success of the system and its exceptional performance was detailed in an article published in 1998. In 2004, our laboratory replaced this system with a new third-generation robotics system, the LH 1502 from Beckman Coulter, Inc. The new system consists of 2 LH 755 workcells (LH 750 plus slide maker/stainer), an inlet/outlet unit, and a stock-yard. The system has been interfaced to our laboratory computer system (Cerner Millieum) by customized software that allows auto-verification and testing rules to be applied to individual samples. Since its implementation in January 2005, we have monitored its performance characteristics relative to the previous first generation system. We report here on our findings through June 2005. We compared and contrasted the two systems with respects to the following parameters: (1) sample handling; (2) reduction in staff exposure to hazardous materials; (3) stat and routine turnaround times; (4) productivity; (5) reduction in backup testing; (6) operating costs; (7) payback; and (8) reduction in FTEs. We found that manual sample handling was virtually identical between the two systems. The LH 1502 holds a slight edge over our older system with respect to staff exposure mainly due to further reduction in manual backup testing. Stat TAT after introduction of the LH 1502 showed an additional 44% drop from 50 to 28 minutes, while the routine TAT was reduced by 23%, down from 61 to 47 minutes. Gains achieved in productivity after installation of the first-generation system were maintained with the LH 1502, with significant extra volume capacity yet to be utilized. The space required for operating the system was also reduced by nearly 49%. There was a 20.3% reduction in backup blood smears that required manual review (due primarily to the lower flagging rates for the LH versus STKS). The improvements in overall operating costs (> dollar 195,000) were also maintained. Payback was not considered since this was a replacement of our previous system, which had already eliminated more than 6 FTEs. PMID- 16475476 TI - Performance evaluation of the latest fully automated hematology analyzers in a large, commercial laboratory setting: a 4-way, side-by-side study. AB - Gamma-Dynacare is a Canadian-based community laboratory partnership formed in the mid-1990s through the merger of 3 prominent Ontario medical diagnostic laboratories. Laboratory Corporation of America acquired an interest in the GD partnership in mid 2002. We service more than 10,000 community-based Canadian clinicians, hospital partners, and private clients with an integrated customer focused system that includes specimen collection, transportation, and results reporting services. With more than 1,700 highly qualified medical, technical, and support staff and a network of laboratories, Gamma Dynacare aims to be at the forefront of technological innovation to better service the clinician base and ultimately deliver better patient care. We were looking for a hematology analyzer that would allow. (1) standardization throughout Ontario in our 4 largest sites and (2) better performance to effectively handle aged samples and minimize slide review. To select the best, most productive hematology analyzer for our environment, it was decided to perform a side-by-side comparison of the top hematology analyzers from Abbott (Cell-Dyn 3500), Beckman Coulter (LH 750), Bayer (Advia 120), and Sysmex (XE 2100), utilizing the same samples. CBC, differential and reticulocyte parameters were all evaluated according to CLSI (formerly NCCLS) and established hematology analyzer evaluation guidelines. We assessed each analyzer for precision, linearity, carryover, stability, differential capabilities, slide review rates, and throughput (clean bench studies). Two hundred samples were assessed for differential and morphology flagging on each analyzer using the reference 400 cell manual differential for comparison. Throughput was assessed by analyzing 700 consecutive samples representative of our workload mix. Stability studies at 24 hours showed that the Beckman Coulter LH 750 was least affected by EDTA, effect with minimal changes in the mean corpuscular volume (MCV) and hematocrit. Both the Bayer Advia 120 and Sysmex XE 2100 showed an elevation of the MCV (up to 5 fL) and Hematocrit over the 24 hours. Analysis of the 200 randomly selected patient samples showed that, while the false-negative rates on each of the instruments were comparable, there were significant differences in the false-positive rates. This has important implications for slide review rates. For our specimen mix, the Sysmex XE 2100 had the highest false-positive rate (15%), followed by the Cell-Dyn 3500 (8%), Advia 120 (6.5%) and the Beckman Coulter LH 750 (1.5%). Reticulocyte analysis performance was observed to be satisfactory with the Beckman Coulter LH 750, Cell Dyn 3500, and XE 2100, while the Bayer Advia 120 showed a decrease in retic values after 12 hours. In conclusion, many laboratories will not be able to perform a 4-way evaluation such as described here due to time, space, and resource constraints. For our laboratories, result quality, sample stability performance, slide review rates, and efficiency were the primary criteria in selection of the most suitable hematology analyzer. Our 4-way evaluation resulted in selection of the Beckman Coulter LH 750 for Gamma-Dynacare Laboratories because it enabled the lowest slide review rate and handled aged samples better than the other analyzers. PMID- 16475478 TI - Sleep microarchitecture in childhood and adolescent depression: temporal coherence. AB - Previous work has indicated that low temporal coherence of ultradian sleep EEG rhythms is characteristic of depressed patients and women in particular. It may also be evident in depressed children and adolescents, although most published studies are limited in sample size. The present study evaluated temporal coherence of sleep EEG rhythms in 173 children and adolescents 8-17 years of age, including 97 who met criteria for major depressive disorder (MDD) and were symptomatic but unmedicated at the time of study and 76 healthy controls. Temporal coherence of all-night sleep EEG rhythms was evaluated on the second of two nights in the laboratory. Data were coded for diagnostic group, gender and age and subjected to MANOVAs. Temporal coherence was significantly lower in adolescents with MDD, compared to healthy controls. Findings were most robust for coherence between left and right beta and between delta and beta in both hemispheres. Both gender and age strongly influenced between-group differences, with the lowest temporal coherence among MDD girls, even in those under 13 years of age. In conclusion, early onset depression is associated with a reduction in synchronization of sleep EEG rhythms that shows a differential maturational course in boys and girls. PMID- 16475479 TI - Functional connectivity between hemispheres and schizophrenic symptoms: a longitudinal study of interhemispheric EEG coherence in patients with acute exacerbations of schizophrenia. AB - To clarify whether interhemispheric electroencephalogram (EEG) coherence reflecting functional connectivity between the two cerebral hemispheres can change in a symptom-dependent manner in schizophrenia, we measured resting EEG and symptom severity twice at an average interval of 32.7 days during the course of treatment in 15 patients hospitalized for acute exacerbations of schizophrenia. Symptom severity was estimated quantitatively by means of the Brief Psychiatric Rating Scale (BPRS). Correlation analysis showed that increases in the beta-band coherence for frontal electrode pairs during the treatment were associated with improvement in the total score and the score on the positive subscale of BPRS. This result suggests that functional disconnection between the left and right frontal lobes may be related to the generation of psychotic symptoms and can normalize following antipsychotic treatment. PMID- 16475480 TI - Frontal alpha power asymmetry in aggressive children and adolescents with mood and disruptive behavior disorders. AB - Building on prior research, which has suggested a relationship between aggression and left frontal activity, our study tested the hypothesis that proneness to impulsive aggression would be related to relative left frontal overactivation. EEG one-hertz resting alpha power frontal asymmetry was examined in 65 pediatric male psychiatric patients with a history of impulsive aggression and comorbid mood and disruptive behavior disorders. The strongest finding, which emerged from this analysis, was a finding of relative increases in left frontal activity compared with right frontal activity. The results also indicated that greater left frontal activity correlated positively with the severity of psychiatric disturbance. These findings suggest that relative increases in left frontal activity may be related to a locus of neurophysiological disruption associated with psychopathology characterized by behavioral and affective disinhibition. Results are discussed within a model of behavioral inhibition system-behavioral activation system theory. PMID- 16475481 TI - Latency differences of N20, P40/N60, P100/N140 SEP components after stimulation of proximal and distal sites of the median nerve. AB - The latencies of SEP N20, P40, N60, P100, and N140 components were measured after stimulation of two different sites, and the differences in relation to conduction velocity and their central functions are discussed. Subjects were 8 healthy right handed males (age 22-31 years, height 164-184 cm). An electrical pulse of 200micro sec duration with an intensity of 2 times the motor threshold was delivered to the wrist and to the elbow alternately at a random rate of 0.1 to 0.3 Hz. Recording electrodes were Cz', C3', and C4' referenced to linked ears. Analysis time was 50 msec before and 450 msec after the stimulus. The band pass was 0.5 Hz to 2 kHz. Subjects were asked to mentally count the number of stimuli. The averaging was interrupted after every 16 to 24 stimuli and checked to determine whether the subject was attentive to the stimuli by verifying the number of stimuli counted. A total of 100 responses each from elbow and wrist stimuli were averaged. Differences in peak latency between elbow and wrist stimuli for N20, P40, N60, P100, and N140 were 3.7 +/- 0.7 msec, 5.0 +/- 1.8, 4.3 +/- 1.2, 8.1 +/- 6.3, and 7.4 +/- 2.6 msec, respectively. According to the latency differences, SEP components can be divided into 3 groups: the shortest difference for N20, medium difference for P40 and N60 and longest difference for P100 and N140. Similar latency differences and similar potential distribution for P40 and N60, and for P100 and N140, and their differences from N20 confirmed that each of N20, P40/N60, and P100/N140 has a different function centrally. In addition, central processing time was longer with the more distal site stimulation. PMID- 16475482 TI - Screening of EEG referrals by neurologists leads to improved healthcare resource utilization. AB - The objective of this study was to determine if screening by a neurologist of all non-neurologist electroencephalogram (EEG) referrals prior to approval reduces the number of inappropriate requests. This retrospective survey included 600 consecutive EEG requisitions referred to the Anaheim Kaiser Permanente Neurodiagnostic Laboratory to rule out epilepsy. Patients with established epilepsy referred for a repeat EEG for management issues were excluded. Three groups of EEG referrals were analyzed. Each group consisted of 200 EEGs (100 pediatric and 100 adult EEGs). The first group was referred directly by non neurologists, the second group was referred by non-neurologists with scrutiny by a neurologist, and the third group was referred by a neurologist directly. In the pediatric group, the ratio of abnormal EEG vs normal EEG was 1:3.35 in the first group, 1:0.69 in the second group and 1:0.33 in the third group. In the adult group, the ratio of abnormal EEGs vs normal EEGs was 1:2.23 in the first group, 1:0.82 in the second group and 1:0.45 in the third group. In the combined pediatric and adult groups, the ratio of abnormal EEG vs normal EEG was 1:2.70 in the first group, 1:0.75 in the second group and 1:0.39 in the third group. There was a significant difference between the results of the EEGs ordered by non neurologists directly versus non-neurologists with scrutiny (p=.334, chi-square test). Scrutiny by a neurologist of EEG referrals from non-neurologists led to a reduction in the number of normal EEG results. This suggests that inappropriate EEG requests for non-epileptic patients that yield normal EEG results are significantly reduced with scrutiny. This can help reduce the indiscriminate overuse of EEGs by non-neurologists thereby leading to better utilization of healthcare resources. PMID- 16475483 TI - Quantitative EEG analysis before and after liver transplantation. AB - Quantitative EEG has been shown to be effective for the assessment of hepatic encephalopathy. Initial quantitative EEG studies of patients who had undergone liver transplantation demonstrated improvement of frequency and the alpha-theta power ratio in the occipital area. This study involved the assessment of comprehensive quantitative EEG variables over the entire cortex following transplantation. Fourteen subjects underwent EEG recording prior to transplantation. Eight (6 without complications and 2 having problems with rejection) underwent the same recording between 3 and 6 months following transplantation. For all subjects, EEG variables showing significant changes from pre- to post-transplantation included posterior alpha frequency (increase, p < or = .03), central theta absolute power (decrease, p < or = .03), theta relative power over anterior, central, and posterior regions (decrease, p < or = .02, p < or = .01, p < or = .03, respectively), posterior beta absolute power (increase, p < or = .01), and central and posterior beta relative power (increase, p < or = .04, p < or = .04, respectively). When the two subjects with complications were removed from the analyses, these variables and also anterior alpha absolute power (increase, p < or = .02), alpha relative power over anterior, central, and posterior regions (increase, p < or = .05, p < or = .03, p < or = .04, respectively), and anterior and central theta absolute power (decrease, p < or = .05, p < or = .04, respectively) showed significant pre- to post-transplant changes. In conclusion, a combination of quantitative EEG parameters which are most affected by liver transplantation might provide an effective assessment tool for determining and quantitatively monitoring the cerebral status of post transplant patients. PMID- 16475484 TI - Ictal nystagmus in a newborn baby after birth asphyxia. AB - Ictal nystagmus (IN) is an uncommon phenomenon characterized by rhythmic saccadic eye movements occurring during epileptic seizures. We report a newborn baby with severe birth asphyxia, undergoing long-term video EEG monitoring with electro oculogram (EOG), who showed irregular IN when eye movements crossed the midline from left to right and vice versa, resulting in large amplitude of the nystagmoid movements. The nystagmus was followed 15 to 29 seconds later by ictal discharges in the occipital regions. MRI of the brain showed features suggestive of periventricular leukomalacia. This interesting combination of findings suggests a complex mechanism for IN of cortical or subcortical ictal rhythms, which results in (a) the generation of subcortical electrical discharges in the pons and midbrain, causing nystagmoid eye movements, and (b) subsequent occipital spiking. We conclude that this clinical manifestation supports the existence of functioning cortical-subcortical connections between the brainstem ocular motor centers and the occipital cortex at birth. PMID- 16475485 TI - Focal seizures originating from the left temporal lobe in a case with chorea acanthocytosis. AB - Neuroacanthocytosis is a group of disorders, clinically characterized with movement disorders, self-mutilation and seizures. There is little information in the literature regarding the clinical and EEG findings of the accompanying seizures in this neurodegenerative disorder. A 46-year-old man who was diagnosed as chorea-acanthocytosis, a subgroup of neuroacanthocytosis, was investigated for severe drug resistant seizures. Continuous video-EEG monitoring revealed an active left temporal epileptogenic focus with left temporal rhythmic discharges detected during his complex partial seizures. T2-weighted cranial MRI indicated hyperintensity of the left amygdala as well as hyperintensity and atrophy of bilateral basal ganglia. We discuss the development of focal seizures in a patient who has a neurodegenerative disease, namely neuroacanthocytosis. PMID- 16475486 TI - Necrotizing leukoencephalopathy associated with nonconvulsive status epilepticus and periodic short-interval diffuse discharges: a clinicopathological study. AB - We describe the clinical, neuroimaging and neuropathological features of an immunocompromised patient diagnosed as having refractory nonconvulsive status epilepticus (NCSE), and whose consecutive electroencephalograms (EEGs) revealed persistent periodic short-interval diffuse discharges (PSIDDs). Prominent subcortical white matter lesions in keeping with the diagnosis of multifocal necrotizing leukoencephalopathy may be neuropathological substrate of NCSE with persistent PSIDDs. PMID- 16475487 TI - QEEG monitoring of Alzheimer's disease treatment: a preliminary report of three case studies. AB - Previous research has shown that quantitative electroencephalography (qEEG) can monitor treatment of Alzheimer's Disease (AD). This study investigated the ability of a qEEG measure based on EEG variance, combined with a delayed recognition memory task, to measure treatment effects on patients with AD. Three AD patients with very mild AD (CDR=0.5, FAST stage 3) were monitored with task specific EEG at multiple time points before and after medication treatment. Patients had their EEG recorded while performing a recognition memory task. A measure of (normalized) variance was applied to the EEG data. To the extent possible, the subjects received this treatment monitoring multiple times. These patients were monitored a total of 14 times, which yielded 11 measurements of qEEG change during the course of treatment. The direction of change in patients' qEEG values agreed with patients' medication treatment on 10 out of 11 occasions, p < 0.006 (binomial test) and was more accurate than monitoring with the relative theta power, p < 0.05. The results of this monitoring also showed that the qEEG measure accurately reflected treatment in a dose dependent manner. These results were independent of the specific medication monitored; Galantamine, Memantine, Nicotine, and Rivastigmine. In conclusion, this qEEG method may be useful for measuring AD treatment responses. PMID- 16475488 TI - Melanoma of childhood and adolescence. PMID- 16475489 TI - Alabaster skin after CO2 laser resurfacing: evidence for suppressed melanogenesis rather than just melanocyte destruction. AB - Hypopigmentation is a recognized side effect of CO2 laser resurfacing. Pigment abnormalities are a major side effect of facial laser procedures and can cause much emotional distress. We report a case of a patient who, after receiving laser treatment, developed persistent hypopigmentation that has defied a variety of treatment attempts. Results of histologic and immunohistochemical studies support the hypothesis that suppressed melanogenesis rather than just destruction of melanocytes is important in the etiology of the alabaster skin side effect. PMID- 16475490 TI - Syringomatous carcinoma in a young patient treated with Mohs micrographic surgery. AB - Syringomatous carcinoma (SC) has typically been observed in middle-aged and older patients. We report a case of SC mimicking an epidermoid cyst in a 23-year-old Asian man. Histopathologic examination results showed a dermal neoplasm consisting of nests of basaloid cells, focal areas of ductal differentiation, moderate dermal fibrosis, and moderate nuclear atypia consistent with a diagnosis of SC. No perineural involvement was noted. Results of 2005. immunohistochemical analysis revealed positivity for high- and low-molecular-weight cytokeratins, as well as for carcinoembryonic antigen (CEA). There was scattered immunoreactivity to S-100 protein. The tumor was completely excised Pennsylvania. using Mohs micrographic surgery (MS). This case demonstrates the importance of differentiating SC from other benign or malignant entities, the value of a prompt diagnosis of SC, and the effective treatment of SC with MMS. PMID- 16475491 TI - Cutaneous lupoid leishmaniasis: a case report. AB - Cutaneous leishmaniasis is a disease transmitted by the sandfly. During the course of the disease, all classical stages of the development of leishmaniasis from small erythematous papules to nodules to ulcerative lesions can be seen. We report a case of lupoid leishmaniasis (LL) treated with daily intramuscular injections of meglumine antimoniate for 20 days with marked improvement of clinical features. PMID- 16475492 TI - Unilateral mediothoracic exanthem: a variant of unilateral laterothoracic exanthem. AB - We report the cases of a 4-year-old Chinese boy and a 33-year-old Chinese man with prodromal constitutional symptoms followed by eruption of small, painless, erythematous, papular lesions on relatively medial aspects of the anterior chest but not as far as the periflexural regions. Spontaneous remission was seen 2 and 3 weeks after rash onset in the child and the adult, respectively. We believe that the terms unilateral mediothoracic exanthem and unilateral thoracic exanthem are appropriate diagnostic labels. Such labels carry clinical significance as the patients can be reassured that although the rash may persist for weeks, final spontaneous remission with no complication is highly likely. Moreover, the risk of contagiousness is low, and we did not insist on isolation for prolonged periods. PMID- 16475493 TI - Acral syringomas presenting as a photosensitive papular eruption. AB - A 43-year-old woman presented with a recurrent and remitting eruption of reddish brown papules in the sun-exposed areas of the arms and forearms that had been present for 2 years. Results of a histopathologic examination revealed syringoma. To the best of our knowledge, this is the first case report of syringomas confined to sun-exposed areas with a recurrent and remitting course over time. We also review the literature on acral syringomas. PMID- 16475494 TI - Cupric keratosis: green seborrheic keratoses secondary to external copper exposure. AB - Exogenous copper is a well-known cause of blue-green dyspigmentation of the hair, nails, and skin. We report the case of a patient with a blue-green discoloration of multiple seborrheic keratoses on his chest and back after swimming in a pool for rehabilitation of a back injury. Metallic analysis using atomic absorption spectrometry was performed on water samples from both the swimming pool and the hot tub the patient was using. Our dermatology clinic tap water was used as a control. Results of this analysis revealed an elevated level of copper in the swimming pool (2.750 ppm) and normal levels in the hot tub (0.502 ppm) and the control sample (0.891 ppm). The copper level in the swimming pool was more than double the recommended maximum set forth by the US Environmental Protection Agency. After the patient discontinued water exercises in the swimming pool, the green discoloration of the seborrheic keratoses disappeared rapidly. We believe this case represents the first report in the literature of the discoloration of epidermal growths secondary to exogenous heavy metal exposure. PMID- 16475495 TI - Blastic natural killer-cell lymphoma: a case report. AB - This article presents a case of blastic natural killer (NK)-cell lymphoma. This rare neoplasm is defined by its clinical presentation, histologic characteristics, and immunostaining pattern. Most cases, including the one presented here, follow an aggressive clinical course. PMID- 16475496 TI - Tazarotene cream for postinflammatory hyperpigmentation and acne vulgaris in darker skin: a double-blind, randomized, vehicle-controlled study. AB - Previous investigations have reported the efficacy of tazarotene 0.1% cream for the treatment of dyschromia associated with photoaging and for acne vulgaris. The present investigation assessed tazarotene 0.1% cream for the treatment of postinflammatory hyperpigmentation (PIH) in a double-blind, randomized, vehicle controlled study of 74 patients from darker racial ethnic groups who had acne. Once-daily application of tazarotene cream was shown to be effective against PIH, achieving significantly greater reductions compared with vehicle in overall disease severity and in the intensity and area of hyperpigmentation within 18 weeks (P< or =.05). Mean levels of erythema, burning, and peeling were no more than trace in both groups throughout the study, and mean levels of dryness were no more than mild in both groups. In our study, tazarotene cream was effective and well tolerated in the treatment of PIH in patients with darker skin. PMID- 16475497 TI - Metabolites produced by nitrogen-fixing Nostoc species. AB - This paper provides a comprehensive overview of metabolites, including lipids and lipid-like compounds, boron-containing macrocycles, arsenolipids, oligopeptides and amino acid derivatives, produced by cyanobacteria of the genus Nostoc. PMID- 16475498 TI - Post-translational modification(s) and cell distribution of Streptomyces aureofaciens translation elongation factor Tu overproduced in Escherichia coli. AB - We cloned EF-Tu from Streptomyces aureofaciens on a pET plasmid and overproduced it using the T7 RNA polymerase system in Escherichia coli. Streptomyces EF-Tu represented more than 40% of the total cell protein and was stored mostly in inclusion bodies formed apically at both ends of E. coli cells. Analysis of the inclusion bodies by transmission and scanning electron microscopy did not reveal any internal or surface ultrastructures. We developed the method for purification of S. aureofaciens EF-Tu from isolated inclusion bodies based on the ability of the protein to aggregate spontaneously. EF-Tu present in inclusion bodies was not active in GDP binding. Purified protein showed a similar charge heterogeneity as EF-Tu isolated from the mycelium of S. aureofaciens and all of the isoforms reacted with EF-Tu antibodies. All isoforms also reacted with monoclonal antibodies against O-phosphoserine and O-phosphothreonine. PMID- 16475499 TI - Mapping the primary structure of copper/topaquinone-containing methylamine oxidase from Aspergillus niger. AB - The amino acid sequence of methylamine oxidase (MeAO) from the fungus Aspergillus niger was analyzed using mass spectrometry (MS). First, MeAO was characterized by an accurate molar mass of 72.4 kDa of the monomer measured using MALDI-TOF-MS and by a pI value of 5.8 determined by isoelectric focusing. MALDI-TOF-MS revealed a clear peptide mass fingerprint after tryptic digestion, which did not provide any relevant hit when searched against a nonredundant protein database and was different from that of A. niger amine oxidase AO-I. Tandem mass spectrometry with electrospray ionization coupled to liquid chromatography allowed unambiguous reading of six peptide sequences (11-19 amino acids) and seven sequence tags (4 15 amino acids), which were used for MS BLAST homology searching. MeAO was found to be largely homologous to a hypothetical protein AN7641.2 (EMBL/GenBank protein accession code EAA61827) from Aspergillus nidulans FGSC A4 with a theoretical molar mass of 76.46 kDa and pI 6.14, which belongs to the superfamily of copper amine oxidases. The protein AN7641.2 is only little homologous to the amine oxidase AO-I (32% identity, 49 % similarity). PMID- 16475500 TI - Distribution of secreted aspartyl proteinases using a polymerase chain reaction assay with SAP specific primers in Candida albicans isolates. AB - Secreted aspartyl proteinase (Sap) distribution among different C. albicans isolates was determined using SAP-specific primers in polymerase chain reaction (PCR) assay. SAP1, SAP2, and SAP3 were detected in 13 of 40 (32.5%), SAP4 in 38/40 (95%), SAP5 were detected in 30/40 (75%), SAP6 in 23/40 (57.5%) of C. albicans strains isolated from blood cultures. SAP1-SAP3 were detected in 37 of 40 (92.5%), SAP4 were detected in 3/40 (7.5%), SAP5 in 3/40 (7.5%), SAP6 in 5/40 (12.5%) of C. albicans strains isolated from vaginal swab cultures. Sap1, Sap2 and Sap3 isoenzymes were found to be related to the vaginopathic potential of C. albicans; Sap4, Sap5 and Sap6 isoenzymes were found to be correlated with systemic infections. PMID- 16475502 TI - Variability of non-mutualistic filamentous fungi associated with Atta sexdens rubropilosa nests. AB - A survey of the filamentous fungi other than the symbiotic one found in association with Atta sexdens rubropilosa colonies was carried out. Different fungal species (27 taxa) were isolated a few days after treating the workers with toxic baits (sulfluramid; Mirex-S), from 40 laboratory and 20 field nests. Syncephalastrum racemosum (54%) and Escovopsis weberi (21%), Trichoderma harzianum (38%) and Fusarium oxysporum (23%) were the prevalent species in laboratory and field nests, respectively. Acremonium kiliense, Acremonium strictum, E. weberi, F. oxysporum, Fusarium solani, Moniliella suaveolens and T. harzianum were found in both nests' groups. We revealed that many filamentous fungi can co-exist in a dormant state inside the nests of these insects and some of them appear to be tightly associated with this environment. PMID- 16475501 TI - Agrobacterium tumefaciens-mediated transformation of Mucor circinelloides. AB - The Agrobacterium tumefaciens-mediated transformation of the zygomycetous fungus Mucor circinelloides is described. A method was also developed for the hygromycin B-based selection of Mucor transformants. Transformation with the hygromycin B phosphotransferase gene of Escherichia coli controlled by the heterologous Aspergillus nidulans trpC promoter resulted in hygromycin B-resistant clones. The presence of the hygromycin resistance gene in the genome of the transformants was verified by polymerase chain reaction and Southern hybridization: the latter analyses revealed integrations in the host genome at different sites in different transformants. The stability of transformants remained questionable during the latter analyses. PMID- 16475503 TI - Growth strategy of heterotrophic bacterial population along successional sequence on spoil of brown coal colliery substrate. AB - The bacterial population of brown coal colliery spoil (Sokolov coal mining district, Czechia) was characterized by measuring viable bacterial biomass, the culturable to total cell ratio (C : T), colony-forming curve (CFC) analysis and species and/or biotype diversity. Bacterial representatives that differed in colony-forming growth (fast and/or slow growers) were used for growth-strategy investigation of heterotrophic bacteria. Spoil substrates from the surface (0-50 mm) and the mineral (100-150 mm) layers were sampled on 4 sites undergoing spontaneous succession corresponding to 1, 11, 21 and 43 years after deposition (initial, early, mid and late stages). The bacterial biomass of the surface layer increased during the initial and early stages with a maximum at mid stage and stabilized in the late stage while mineral layer biomass increased throughout the succession. The maxima of C : T ratios were at the early stage, minima at the late stage. Depending on the succession stage the C : T ratio was 1.5-2 times higher in the mineral than the surface layer of soil. An increase in the fraction of nonculturable bacteria was associated with the late succession stage. CFC analysis of the surface layer during a 3-d incubation revealed that the early succession substrate contained more (75%) rapidly colonizing bacteria (opportunists, r-strategists) than successively older substrates. The culturable bacterial community of the mineral layer maintained a high genera and species richness of fast growers along the succession line in contrast to the surface layer community, where there was a maximum in the abundance of fast growers in the early stage. There was a balanced distribution of Gram-positive and Gram negative representatives of fast growers in both layers. A markedly lower abundance of slow growers was observed in the mineral in contrast to the surface layer. Gram-positive species dominated the slow growers at the surface as well as in the mineral layers. The growth strategy of the heterotrophic bacterial population along four successional stages on spoil of brown coal colliery substrate in the surface layer displayed a trend indicative of a r-K continuum in contrast to the mineral layer, where an r-strategy persisted. PMID- 16475504 TI - Use of green fluorescent protein as molecular marker for tagging Bacillus brevis in soil under the control of a novel constitutive promoter F1. AB - A constitutive expression vector pHY300-Flgfp was constructed to test the function of promoter F1 subcloned from a rice epiphyte Bacillus brevis strain DX01. The DX01 cells harboring plasmid pHY300-F1gfp were detected to produce bright green fluorescence. Subsequently, the gfp-tagged B. brevis strain was released into the soil and its survival was investigated by PCR and the detection of green fluorescence. The spatial location of in situ gfp-tagged bacterial cells on the root surface of rice seedlings was visualized. All these results indicated that green fluorescent protein is an ideal molecular marker for the detection of the activities of promoter F1, and it is also a reliable probe to monitor specific B. brevis bacteria in the environment. PMID- 16475505 TI - Intramucosal bacteria in colon cancer and their elimination by probiotic strain Enterococcus faecium M-74 with organic selenium. AB - Intraepithelial bacteria were isolated by the gentamicin protection assay (GPA) from biopsy samples obtained at colonoscopy (colon cancer, n = 10 patients; colonic adenoma, n = 20; control group, n = 20; cancer patients without gastrointestinal tract GIT malignancy, n = 10). After a three-month administration of E. faecium M-74 to patients with positive GPA biopsies, 172 biopsy specimens from 60 patients were examined with the GPA. The number of biopsies with intracellular bacteria was significantly higher in adenoma and carcinoma group than in control group (26 vs. 10%; p = 0.004); in cancer patients without GIT malignancy the difference was nonsignificant. E. faecium M-74 was also administered to 5 patients with colonic adenoma; according to a control colonoscopy the number of biopsies with intracellular bacteria was significantly lower after probiotic administration (48 vs. 16%; p = 0.03). A striking prevalence of intraepithelial bacteria was also showed in patients with large bowel adenoma and carcinoma. The administration of probiotic strain M-74 can thus be considered to be an effective and promising method for elimination of pathogenic bacteria in the case of inflammatory bowel disease and colon cancer. PMID- 16475506 TI - Direct immunofluorescence assay for rapid environmental detection of Vibrio cholerae O1. AB - An immunofluorescence assay for direct detection of V. cholerae O1 was developed using polyclonal antibodies raised against outer membrane proteins (OMPs) of V. cholerae O1. Production of OMPs varied with growth media used; maximum production was found in tryptic soy broth. The detection system was specific because no cross-reactivity was observed with other bacteria including V. cholerae O139, E. coli, S. dysenteriae and Salmonella enterica subsp. enterica serovar Typhi. The technique was able to detect 240 CFU/mL of V. cholerae O1 suspended in phosphate buffered saline. The assay coupled with bacterial enrichment in APW for 6 h detected as few as 5 CFU of V. cholerae in spiked samples. Moreover, a 2-h incubation of enriched bacterial cells in 0.1% yeast extract with 10 ppm nalidixic acid enhanced the bacterial size and helped in morphological identification of V. cholerae. Among 32 potable water samples from afflicted hand pumps and wells collected from a cholera-plagued area 12 were found to be contaminated with V. cholerae by immunofluorescence assay as well as by conventional culture methods. The proposed method could thus be employed in environmental surveillance of V. cholerae O1. PMID- 16475507 TI - Invasive meningococcal disease and latex agglutination test--is it still beneficial for diagnosis? AB - We showed current clinical usefulness of the latex agglutination (LA) test for confirmation of meningococcal etiology on 32 cerebrospinal fluid, 77 serum and 93 urine samples collected during the first week of hospitalization from 19 patients with laboratory confirmed invasive meningococcal disease. The positivity of the LA test in cerebrospinal fluid was 47%, in serum 42% and in urine 24%, while the PCR of cerebrospinal fluid and serum was positive in 95 and 47% cases, respectively. The latest positivity of the LA test was detected on day 2 in cerebrospinal fluid, on day 3 in serum and on day 4 in urine. In the group of patients who had received antibiotic therapy we found nonsignificant reduction of LA positivity and also statistically significant reduction of culture positivity in CSF (p = 0.04); the PCR positivity changed minimally. In blood samples, nonsignificant reduction of culture positivity and no difference in LA and PCR positivity was found. We did not find any statistically significant relationship between test results and clinical forms. The LA test can be therefore considered to be an auxiliary diagnostic method, rapid and easily practicable but less sensitive than PCR. It can be recommended especially for local laboratories where PCR is not available and the patient already received antibiotics before admission to the hospital. PMID- 16475508 TI - Co-inoculation of Borrelia afzelii with tick salivary gland extract influences distribution of immunocompetent cells in the skin and lymph nodes of mice. AB - The impact of Ixodes ricinus salivary gland extract (SGE) on inflammatory changes in the skin and draining lymph nodes of mice, elicited by the infection with the important human pathogen, B. afzelii, was determined using flow cytometry. SGE injected together with spirochetes reduced the numbers of leukocytes and gammadelta-T lymphocytes in infected epidermis at early time-points post infection. In draining lymph nodes, the anti-inflammatory effect of SGE was manifested by the decrease of total cell count compared with that in mice treated with inactivated SGE. Changes in subpopulations of immunocompetent cells apparently reflected the effect of SGE on the proliferation of spirochetes in the host. The significance of tick saliva anti-inflammatory effect for saliva activated transmission of B. afzelii is shown. PMID- 16475509 TI - Diagnosis of transmissible spongiform encephalopathies in animals: a review. AB - Transmissible spongiform encephalopathies (TSEs) in animals include, among others, bovine spongiform encephalopathy (BSE), scrapie, chronic wasting disease, and atypical forms of prion diseases. Diagnosis of TSEs is based on identification of characteristic lesions or on detection of the abnormal prion proteins in tissues, often by use of their partial proteinase K resistance property. Correctly sampling of target tissues is of utmost importance as this has a considerable effect on test sensitivity. Most of the rapid or screening tests are based on ELISA or Western immunoblot (WB) analysis, and many are officially approved. Confirmatory testing is normally performed by use of histologic examination, immunohistochemical analysis, certain WB protocols, or detection of prion fibrils by use of electron microscopy (scrapie-associated fibril). The discriminatory methods for diagnostic use are mostly based on WB technology and provide initial identification of the prion strain, particularly for differentiation of BSE from scrapie in small ruminants. Definitive prion strain characterization is performed by use of bioassays, usually in mice. A burgeoning number of transgenic mice have been developed for TSE studies. Development of new tests with higher sensitivity and of more reliable diagnostic applications for live animals tested for food safety reasons is a rapidly developing field. Ultimately, the choice of a test for TSE diagnosis depends on the rationale for the testing. PMID- 16475510 TI - Clostridial enteric infections in pigs. AB - Clostridium perfringens types A and C and Clostridium difficile are the principal enteric clostridial pathogens of swine. History, clinical signs of disease, and gross and microscopic findings form the basis for a presumptive diagnosis of C. perfringens type-C enteritis. Confirmation is based on isolation of large numbers of type-C C. perfringens and/or detection of beta toxin in intestinal contents. Diagnosis of C. perfringens type-A infection, however, remains controversial, mostly because the condition has not been well defined and because type-A organisms and their most important major (alpha) toxin can be found in intestinal contents of healthy and diseased pigs. Isolation of large numbers of C. perfringens type A from intestinal contents, in the absence of other enteric pathogens, is the most reliable criterion on which to base a diagnosis. Recently, beta2 (CPB2) toxin-producing C. perfringens type A has been linked to disease in piglets and other animals. However, implication of CPB2 in pathogenesis of porcine infections is based principally on isolation of C. perfringens carrying cpb2, the gene encoding CPB2, and the specific role of CPB2 in enteric disease of pigs remains to be fully defined. Clostridium difficile can also be a normal inhabitant of the intestine of healthy pigs, and diagnosis of enteric infection with this microorganism is based on detection of its toxins in feces or intestinal contents. PMID- 16475511 TI - Development of a real-time PCR for detection of Mycoplasma bovis in bovine milk and lung samples. AB - A real-time polymerase chain reaction (PCR) assay using hybridization probes on a LightCycler platform was developed for detection of Mycoplasma bovis from individual bovine mastitis milk and pneumonic lung tissues. The detection limit was 550 colony forming units (cfu)/ml of milk and 650 cfu/25 mg of lung tissue. A panel of bovine Mycoplasma and of other bovine-origin bacteria were tested; only M. bovis strains were positive, with a melting peak of 66.6 degrees C. Mycoplasma agalactiae PG2 was also positive and could be distinguished because it had a melting peak of 63.1 degrees C. In validation testing of clinical samples, the relative sensitivity and specificity were 100% and 99.3% for individual milks and 96.6% and 100% for the lung tissue. Using M. bovis real-time PCR, the M. bovis culture-positive milk samples were estimated to contain between 5 x 10(4) and 7.7 x 10(8) cfu/ml and the M. bovis culture-positive lungs between 1 x 10(3) and 1 x 10(9) cfu/25 mg. Isolation, confirmed with the real-time PCR and colony fluorescent antibody test, showed that at the herd level, the proportion of samples positive for M. bovis isolation in mastitis milk samples submitted to the Mastitis Laboratory, Animal Health Laboratory, University of Guelph, Ontario, Canada, was 2.4% (5/201). We conclude that this probe-based real-time PCR assay is a sensitive, specific, and rapid method to identify M. bovis infection in bovine milk and pneumonic lungs. PMID- 16475512 TI - Status spongiosus of white matter in newborn Gelbvieh-cross calves. AB - Various forms of status spongiosus occur in neonatal cattle, the best characterized of which is due to mutations of the branched-chain alpha-keto acid dehydrogenase complex (BCKD), resulting in bovine maple syrup urine disease (MSUD, branched-chain ketoaciduria). A distinctive neurological syndrome was identified between 1998 and 2003 in 9 calves in a 250-cow stabilized Gelbvieh-Red Angus herd. Both sexes were affected (6 heifers, 3 bulls), with a low annual incidence (3 cases in 1998; no cases in 1999; 2 cases in 2000; 2 in 2001; 1 in 2002; 1 in 2003). Affected calves were born full-term, unable to stand, and had constant whole-body tremors when stimulated. Animals remained in lateral recumbency until death or euthanasia; the longest survival time was 10 days postpartum. The principal histological change in 2 affected calves was diffuse, moderately severe bilaterally symmetrical status spongiosus with Alzheimer type II cells throughout the white matter of the brain. Myelin deficits were not evident and vacuoles were due to cleaved myelin sheaths. Neither recognized mutation of MSUD was identified in the E1a subunit of BCKD in 2 affected calves, 8 dams that gave birth to affected calves, a grand-dam of 3 affected calves, or a sire of 1 calf. Amino acid analysis of serum from 1 affected calf revealed normal concentrations of branched-chain amino acids, indicating that this disease is distinct from MSUD. The genetic and biochemical basis for the disorder, provisionally named congenital status spongiosus of Gelbvieh-cross cattle, is undetermined. The pattern of inheritance was not established. PMID- 16475513 TI - Effect of egg yolk on the detection of Mycobacterium avium subsp. paratuberculosis using the ESP II liquid culture system. AB - Rapid diagnosis of paratuberculosis in infected cattle is important for the successful control of Johne disease within herds. Thus, improving culture methods for Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) will aid in the identification of asymptomatic animals. Egg yolk is a component of the media used for growing M. paratuberculosis, but its requirement as a supplement has not been reported. Using the ESP II liquid culture system, 2 different sources and 5 concentrations (3.3%, 1.6%, 0.8%, 0.4%, and 0%) of egg yolk were analyzed. Egg yolk source did not affect either recovery rate or time to detection, but both parameters were significantly improved when the 3.3% egg yolk concentrations (final volume) were used over media containing no egg yolk. This study also assessed the recovery of M. paratuberculosis from fecal samples that were cultured multiple times using Herrold egg yolk agar (HEY). Specimens containing greater than 70 cfu/g feces could routinely be identified as positive for M. paratuberculosis after only 1 culture attempt, whereas specimens with fewer bacteria were only intermittently positive, even after 5 replicate cultures. Therefore, this study indicates that the sensitivity of the Trek Diagnostic ESP II liquid culture system for M. paratuberculosis is affected by egg yolk concentration and that single culture attempts using HEY solid media may not identify specimens containing low numbers of bacteria. PMID- 16475514 TI - Variation among pathologists in the histologic grading of canine cutaneous mast cell tumors with uniform use of a single grading reference. AB - Ten veterinary pathologists independently assigned histologic grades to the same 60 canine cutaneous mast cell tumors using the Patnaik classifications. The degree of agreement in grading among the pathologists was compared with the degree of agreement among the same pathologists in a previous study, in which each pathologist used the reference for grading that he/she uses routinely. Mean agreement improved significantly from 50.3% to 62.1% with uniform use of the Patnaik classifications (P = 0.00001), suggesting that there is value in uniform application of a single grading scheme for canine cutaneous mast cell tumors. Agreement among pathologists was still not 100%, suggesting that a more objective grading scheme should be developed and that other histologic indicators of prognosis should be investigated. PMID- 16475515 TI - Use of a multiplex polymerase chain reaction to rapidly differentiate Neospora caninum from Toxoplasma gondii in an adult dog with necrotizing myocarditis and myocardial infarct. AB - This report describes a 3-year-old male castrated Mastiff dog that died unexpectedly with locally extensive, acute, necrotizing myocarditis and myocardial infarction. Intralesional protozoal tachyzoites in the affected myocardium were confirmed to be Neospora caninum by a novel multiplex polymerase chain reaction (PCR) and immunohistochemistry. Protozoal organisms were not identified in other tissues by histology, immunohistochemistry, or PCR. The multiplex PCR assay was used to quickly provide preliminary results on fresh myocardium to differentiate N. caninum and Toxoplasma gondii. Neosporosis is an uncommon cause of myocarditis in adult dogs and differential diagnoses for myocarditis in this population of dogs are reviewed. PMID- 16475516 TI - Development of a SYBR green real-time polymerase chain reaction assay for quantitative detection of Babesia gibsoni (Asian genotype) DNA. AB - A real-time fluorogenic polymerase chain reaction (PCR) assay based on SYBR green that allows for sensitive, reproducible, and accurate quantification of Babesia gibsoni (Asian genotype). DNA from peripheral blood of infected dogs was developed. Standard curves were created by plotting the input amount of a standard template, constructed with plasmid DNA containing 182 base pairs (bp) of the p18 gene, against threshold cycle numbers. The curves showed a wide dynamic range (1,000,000-fold input) and high correlation values (>0.99). The PCR amplification efficacy of the standard template was similar to that of intact genomic DNA obtained from peripheral blood with B. gibsoni infection. The detection limit of the assay was 9 parasites/microl of blood with B. gibsoni infection. The intra-assay and interassay coefficients of variation of the threshold cycles ranged from 0.70% to 1.89% and from 1.18% to 1.92%, respectively. This assay system was found to be reproducible and accurate for the quantification of parasite DNA in experimentally infected dogs and far more sensitive than traditional microscopic examination. PMID- 16475517 TI - Comparison of six RNA extraction methods for the detection of classical swine fever virus by real-time and conventional reverse transcription-PCR. AB - Six RNA extraction methods, i.e., RNAqueous kit, Micro-to-midi total RNA purification system, NucleoSpin RNA II, GenElute mammalian total RNA kit, RNeasy mini kit, and TRIzol LS reagent, were evaluated on blood and 7 tissues from pig infected with classical swine fever virus (CSFV). Each of the 6 extraction methods yielded sufficient RNA for positive results in a real-time reverse transcription-PCR (RT-PCR) for CSFV, and all RNA, except the one extracted from blood by TRIzol LS reagent, yielded positive results in both a conventional RT PCR for CSFV and a conventional RT-PCR for an endogenous gene encoding beta actin. The RNA extracted from blood by TRIzol LS reagent became positive in both conventional RT-PCR assays when it was diluted to 1:2, 1:4, or up to 1:64 in nuclease-free water. It is concluded that all 6 methods are more or less useful for the detection of CSFV by real-time and conventional RT-PCR in swine blood and tissues. However, some of the 6 reagents offer certain advantages not common to all 6 extraction procedures. For example, RNA extracted by the TRIzol LS reagent constantly had the highest yield; that by the RNAqueous kit had the highest A260/A280 ratio for almost all samples; and that by the NucleoSpin RNA II and the GenElute mammalian total RNA kit was most likely to be free of contaminations with genomic DNA. PMID- 16475518 TI - Equine herpesvirus-4 kinetics in peripheral blood leukocytes and nasopharyngeal secretions in foals using quantitative real-time TaqMan PCR. AB - Based on the hypothesis that the viral load of cells infected with EHV-4 will likely change during the course of disease, TaqMan PCR was used to investigate and characterize the kinetics of EHV-4 viral DNA load (glycoprotein B gene) and transcriptional activity (glycoprotein B and latency-associated transcripts) in peripheral blood leukocytes (PBLs) and nasopharyngeal secretions (NSs) collected from 11 foals during a field outbreak of respiratory disease. The EHV-4 DNA load in PBLs was low and of short duration after onset of clinical signs. In contrast, the EHV-4 load in NSs remained high for the majority of the foals over a period of 4 weeks. Viral replication determined by detection of mRNA expression of the structural glycoprotein B was detected only in NSs during the first 7 days after onset of clinical signs for most foals. The majority of foals expressed latency associated transcripts in NS sonly during the first 7 days after onset of clinical signs. Persistence of the expression of latency-associated transcripts in NS, as a reflection of a latent viral state, was not documented during the 28 day study period. Based on these results, it was concluded that lytic infection with EHV-4 can be diagnosed either by high EHV-4 DNA load of glycoprotein B gene or by detection of transcriptional activity of glycoprotein B. PMID- 16475519 TI - Cloning and sequencing of equine cardiac troponin I and confirmation of its usefulness as a target analyte for commercial troponin I analyzers. AB - The analysis of cardiac troponin I (cTnI) in the diagnosis of myocardial injury in domestic animals is gaining popularity. In this study, equine cTnI was sequenced and compared with previously characterized cTnI from other species. A 6 amino-acid N-terminal deletion unique to the horse was identified. This deletion was outside the epitope region of cTnI recognized by most commercial immunoassays and did not affect the ability of a commercial analyzer system to detect recombinant equine cTnI. No function could be ascribed to the deleted portion. These data support the use of commercial analyzers in measuring equine cTnI. PMID- 16475520 TI - A method for extracting genomic DNA from individual elaphostrongyline (Nematoda: Protostrongylidae) larvae and differentiation of Elaphostrongylus spp. from Parelaphostrongylus spp. by PCR assay. AB - This article reports a rapid and effective method for the extraction and purification of genomic DNA (gDNA) from individual first-stage larvae (L1) of elaphostrongyline nematodes that had been stored frozen or fixed in 95% ethanol for 1 to 5 years. The method was highly effective for L1s of all 6 species of elaphostrongylines, based on polymerase chain reaction (PCR) amplification of a partial fragment of the first internal transcribed spacer (ITS-1) of the ribosomal DNA. Differences were detected in the sizes of partial ITS-1 amplicons between the 2 elaphostrongyline genera, Elaphostrongylus and Parelaphostrongylus. The reliability of the ITS-1 PCR assay was tested by using L1s of unknown identity from Newfoundland and Labrador, Canada. The ability to consistently isolate gDNA from individual L1s, together with a simple PCR-based method to distinguish between Parelaphostrongylus and Elaphostrongylus, have important implications for diagnostic testing and for conducting epizootiological studies on these parasites of veterinary importance. PMID- 16475521 TI - Bovine viral diarrhea virus in alpaca: abortion and persistent infection. AB - An alpaca herd in eastern Ontario experienced vague signs of illness, including anorexia and lethargy in 9 animals, 2.5 months after the addition of a chronically ill cria and his dam to the farm. Subsequently 2 alpaca had early pregnancy loss; one aborted at 5.5 months gestation and the other at 7 months gestation. Seventeen were found to have serum antibody to bovine viral diarrhea virus (BVDV), with highest titers to BVDV type 1. The fetus that was aborted at 5.5 months gestation, 3 months after the clinical outbreak, was found to be positive for BVDV on immunohistochemical staining, and noncytopathic BVDV type 1b was isolated. Of the 13 cria born alive that season, a single male underweight alpaca cria, born 9 months after the clinical illnesses, was infected with BVDV type 1b. The cria was positive for BVDV at birth, at 3 and 26 days of age and continued to be positive for noncytopathic BVDV using virus isolation, nested reverse transcription PCR, antigen detection ELISA, and immunohistochemical staining until euthanasia at 46 days of age. The cria remained serum antibody negative to both BVDV type 1 and type 2. A diagnosis of persistent infection was made. This is the first report describing persistent infection with BVDV in an alpaca cria. PMID- 16475522 TI - Persistent viral shedding during asymptomatic Aleutian mink disease parvoviral infection in a ferret. AB - A 2-year-old domestic ferret that appeared clinically healthy was repeatedly seropositive for Aleutian mink disease parvovirus (ADV) over a 2-year observation period. Antibody titers, determined by counter-immunoelectrophoresis, ranged from 1024 to 4096. Viral DNA also was identified in serum, urine, feces, and blood cell fractions by polymerase chain reaction analysis. Ultimately, DNA in situ hybridization revealed ADV DNA in histologic sections of various tissues and organs. These data indicate that this asymptomatic ferret was persistently infected with ADV. PMID- 16475523 TI - Systemic Cryptococcus albidus infection in a Doberman Pinscher. AB - Cryptococcus albidus is a saprophytic, encapsulated yeast usually found in air, both outdoor and indoor, and sometimes on human skin. It is not usually considered to be a primary pathogen. Most cryptococcal infections of humans and animals are caused by Cryptococcus neoformans. Several cases of C. albidus infection have been reported in humans over the past 20 years. In the veterinary literature, 2 equine cases have been described: genital infection and mycotic keratitis. The present report is the first documented case of C. albidus systemic infection in a dog. Veterinarians and diagnosticians should be aware that C. albidus may be a potential canine pathogen. PMID- 16475524 TI - Occurrence of Piscirickettsiosis-like syndrome in tilapia in the continental United States. AB - From 2001 to 2003, tilapia (Oreochromis sp.) farms in Florida, California, and South Carolina experienced epizootics of a systemic disease causing mortality. The fish exhibited lethargy, occasional exophthalmia, and skin petechia. The gills were often necrotic, with a patchy white and red appearance. Grossly, the spleen and kidneys were granular with whitish irregular nodules throughout. Granulomatous infiltrates were observed in kidney, spleen, testes, and ovary tissues, but not in the liver. The granulomas contained pleomorphic coccoid bacteria, measuring 0.57 +/- 0.1 x 0.8 +/- 0.2 microm, that were Giemsa-positive, acid-fast-negative, and Gram-negative. The bacteria had a double cell wall, variable electron-dense and -lucent areas, and were present in the cytoplasm and within phagolysosomes. The syndrome was associated with cold stress and poor water conditions. These findings are consistent with an infectious process caused by a Piscirickettsia-like bacterium described previously in tilapia in Taiwan and Hawaii. This report involves the first identified cases of a piscirickettsiosis like syndrome affecting tilapia in the continental United States. PMID- 16475525 TI - Seasonal meningoencephalitis in Holstein cattle caused by Naegleria fowleri. AB - Primary amoebic meningoencephalitis is a fulminant infection of the human central nervous system caused by Naegleria fowleri, a free-living amoeba that thrives in artificially or naturally heated water. The infection usually is acquired while bathing or swimming in such waters. The portal of entry is the olfactory neuroepithelium. This report describes fatal meningoencephalitis caused by N. fowleri in Holstein cattle that consumed untreated surface water in an area of California where summer temperatures at times exceed 42 degrees C. In the summers of 1998 and 1999, severe multifocal necrosuppurative hemorrhagic meningoencephalitis was observed in brain samples from nine 10-20-month-old heifers with clinical histories of acute central nervous system disease. Olfactory lobes and cerebella were most severely affected. Lesions were also evident in periventricular and submeningeal neuropil as well as olfactory nerves. Naegleria fowleri was demonstrated by immunohistochemistry in brain and olfactory nerve lesions and was isolated from one brain. Even though cultures of drinking water did not yield N. fowleri, drinking water was the likely source of the amoeba. The disease in cattle closely resembles primary amoebic meningoencephalitis in humans. Naegleria meningoencephalitis should be included among differential diagnoses of central nervous system disease in cattle during the summer season in areas with high ambient temperatures. PMID- 16475526 TI - Canine coronavirus-associated puppy mortality without evidence of concurrent canine parvovirus infection. AB - This report presents 2 cases in which puppy fatalities were associated with canine coronavirus (CCV), but no evidence of concurrent canine parvovirus (CPV-2) disease was observed. Case 1 involved a 7-week-old, male short-haired Chihuahua, which had become lethargic 24 hours after purchase from a pet store. Within 72 hours, the puppy began to vomit, had diarrhea, and was admitted to the veterinary clinic, where it was placed on IV fluids. The parvovirus Cite test was negative. The puppy died within 12 hours of admission and was submitted for diagnostic workup. Gross pathology revealed an enteritis suggestive of CPV-2. Histopathology on intestines showed scattered dilated crypts with necrotic cellular debris and neutrophils. There was moderate depletion and necrosis of lymphoid follicles. Electron microscopy (EM) on intestinal contents was positive for coronavirus and negative for parvovirus. Immunohistochemistry (IHC) on gut sections was positive for CCV and negative for CPV-2. Case 2 was an 8-week-old, male Shih Tzu, which was admitted to the veterinary clinic exhibiting symptoms of severe gastroenteritis with abdominal pain. The referring veterinarian euthanized the puppy, and the entire body was submitted for diagnostic evaluation. Necropsy revealed a severe ileo-cecal intussusception and segmental necrotic enteritis of the small intestine. Electron microscopy of the intestinal contents was positive for coronavirus and negative for parvovirus. Immunohistochemistry on sections of affected gut were positive for CCV and negative for CPV-2. These cases emphasize the importance of pursuing a diagnosis of CCV in young puppies when CPV-2 disease has been ruled out by IHC. PMID- 16475528 TI - Fatal necrotizing fasciitis and myositis in a captive common bottlenose dolphin (Tursiops truncatus) associated with Streptococcus agalactiae. AB - A common bottlenose dolphin (Tursiops truncatus) was presented for necropsy after acute onset of gastrointestinal signs and cutaneous lesions that rapidly progressed to death. Gross and microscopic findings were characterized by locally extensive severe necrohemorrhagic fasciitis and cellulitis, and severe necrotizing myositis in the head and dorsocranial thorax, with numerous disseminated gram-positive cocci. Streptococcus agalactiae was isolated from the lesions and from visceral organs (liver and lung), and it was identified by standard microbiology techniques. This communication is the first report of necrotizing fasciitis in a marine mammal associated with S. agalactiae. PMID- 16475527 TI - Eastern equine encephalitis in dogs. AB - Eastern equine encephalitis virus (EEEV) is an Alphavirus that is endemic in the Southeastern United States. From 1993 to January 2005, the Veterinary Diagnostic and Investigational Laboratory in Tifton, Georgia, performed postmortem examinations on over 101 domestic canines exhibiting clinical neurological disturbances. In 12 of these dogs, brains were histologically suggestive of infection with EEEV. All dogs were less than 6 months of age, with no breed predilection. Clinical signs included pyrexia, depression, nystagmus, and lateral recumbency. Microscopically, brains from all 12 puppies contained infiltrates of lymphocytes, plasma cells, and histiocytes, with occasional neutrophils and random foci of astrocytosis and gliosis. There were mild to moderate perivascular infiltrates of neutrophils along with scattered lymphocytes, plasma cells, and macrophages in the meninges. Viruses isolated from brain homogenates of all 12 puppies were confirmed by indirect fluorescent antibody testing to be EEEV. Additionally, RNA extracted from the brains and viral cultures of 2 dogs were determined by a specific reverse-transcriptase polymerase chain reaction (RT-PCR) to contain EEEV. The single available serum sample exhibited a 1:8 serum neutralization titer to EEEV. PMID- 16475529 TI - Intrarenal pelvic nephroblastoma in a meerkat (Suricata suricatta). AB - Nephroblastoma is the most common primary renal tumor in children and has also been reported in domestic and nondomestic animal species. Intrapelvic renal nephroblastoma is a rare variant of this tumor type in human patients. Postmortem examination of a captive meerkat (Suricata suricatta), which was found dead, revealed enlargement of the pelvis of the left kidney by a tumor mass. Gross, histological, and immunohistochemical findings were consistent with a diagnosis of triphasic intrapelvic renal nephroblastoma. This is the first reported spontaneous case of intrapelvic renal nephroblastoma in a nonhuman species. PMID- 16475530 TI - Gossypol toxicosis in a dog consequent to ingestion of cottonseed bedding. AB - Six dogs died after accidental ingestion of cottonseed bedding. No clinical signs of illness were observed prior to death. A full diagnostic workup was performed on one of these dogs. At necropsy, the lungs were congested and edematous, and the liver was firm, congested, and had a marked reticular pattern. There was also moderate ascites. Histopathologic examination revealed multifocal myocardial degeneration and necrosis, severe pulmonary edema, and chronic passive congestion of the lungs, heart, liver, and kidneys. Transmission electron microscopy of the myocardium revealed disruption of myofibrils, chromatin condensation, and disrupted and swollen mitochondria. The cottonseed bedding contained 1,600 mg/kg of free gossypol, a concentration considered toxic for monogastric animals. The stomach content revealed the presence of gossypol, thus confirming ingestion of cottonseed. Gossypol poisoning in dogs is extremely rare and has not yet been associated with cottonseed bedding. This first documented case of gossypol poisoning in a dog, caused by the ingestion of cottonseed bedding, demonstrates how specific toxicological analysis is crucial in reaching an accurate diagnosis. PMID- 16475531 TI - Cathy Geddes. PMID- 16475532 TI - Extending the scope of inquiry. PMID- 16475533 TI - Primary care reform: opportunities and threats. PMID- 16475534 TI - Made to measure. PMID- 16475535 TI - Advanced primary nursing: liberating the talents. PMID- 16475536 TI - Why stay in nursing? PMID- 16475537 TI - Learning to lead. PMID- 16475538 TI - [Chondrosarcomas of the larynx: diagnostic and therapeutic controversies]. AB - Chondrosarcomas of the larynx are rare neoplasms that account only for a very small percentage of all the primary laryngeal neoplasms that has been estimated in less than 2%. They often show initially with indolent symptoms despite their progressive growth until great compromise of the laryngeal function is produced. This neoplasm is characterised by a variable local aggressive behaviour and frequent of distant metastasis, that are commonly located in lungs and bone, deppending on the hystologic grade. A case of an aggressive laryngeal chondrosarcoma with exclusive brain metastasic disease as cause of death, a very uncommon event in these neoplasms, is reported. We also provide a review of the literature focusing on the controversies about the management of this condition. PMID- 16475539 TI - [Lateral sinus thrombosis due to cholesteatoma. Report of a case and literature review]. AB - After introduction of antibiotics, endocranial complications in otitis media are less common. Lateral sinus thrombophlebitis (LST) incidence has markedly decreased but mortality is still high. This complication should be considered in patients with ear discharge, fever and neurological symptoms. CT-scan and MRI enables early diagnosis and have a role in detecting addition intracranial complications. Surgical intervention should be aggressive and anticoagulation should be considered. We present a case of a woman with othorrea, fever and neurological symptoms. The CT-scan shows us a cholesteatoma that grows intracranial and a LST as complication of it. PMID- 16475540 TI - [Metastases in both internal auditive meatus of nasosinusal melanoma]. AB - Primary malignant melanoma of the nasal and paranasal sinusses is rare and represent 3% of malignant nasosinusal tumors. The prognosis of this tumor is poor. The evolution is unpredictable; there is very commonly a series of local recurrences which may be well controlled, followed by remote metastases remaining long quiescent. There are no statistical difference in local control or survival between patients receiving surgery alone and those receiving surgery and radiotherapy. Chemotherapy is used for metastatic forms. We present a case of a patient with a primary malignant melanoma of the nasal mucosa. Tree years after diagnosis we found intracranial metastases in left and right auditive internal meatus. PMID- 16475541 TI - [Middle ear effusion as infrequent cause of sensorineural hearing loss]. AB - Middle ear efussion (MEE) is the most frequent condition related to conductive hearing loss in children, being rarely the cause of sensorineural hearing loss (SNHL). We present a 5-years-old child without know previous otologic disease who developed MEE with a very adversely effect on hearing. Hearing tests showed a bilateral moderate-severe neural impariment. The patient were programmed for grommet insertion. Hearing showed a total audiometric recovery three months after surgery. To sum up, MEE can be rarely associated with SNHL. PMID- 16475543 TI - [Recurrent and severe epistaxis secondary to nasal trauma. Report of an atypical case in a 22-year-old male]. AB - We report the case of a 22-year-old male who presented a diferred, recurrent and severe epistaxis after nasal trauma which required several admissions in our hospital and different treatments. Concretely by chronological order it has been practiced anterior tamponades, endoscopic ligature-cauterization of the sphenopalatine artery, classic posterior packing and finally a new FENS with cauterization of anterior ethmoidal artery. Besides it was necessary a transfusion to go up the anemia suffered by the patient whose global stay was 16 days. Recurrent postraumatic epistaxis are very rare but can be represent a difficult problem to solve. PMID- 16475542 TI - [Internal jugular vein spontaneous thrombosis. Clinical case and review of the literature]. AB - Internal jugular vein thrombosis (IJT) represents a potentially serious pathology that can be spontaneous or secondary to head and neck infections, surgery, venous catheterization and intravenous drug abuse among others. We report the case of a male affected with IJT without clear cause after performing a complete exam which included ultrasonography, Doppler and contrast-enhanced CT scanning to confirm the diagnosis. Actually its treatment is mainly mediacl by antibiotic therapy and anticoagulants. Besides, it must be excluded an associated neoplasm. PMID- 16475544 TI - [Acute palsy of twelfth cranial nerve]. AB - The hypoglossal nerve or Twelfth-nerve palsy is a rare damage with different causes: tumors or metastases in skull base, cervicals tumors, schwannoma, dissection or aneurysm carotid arteries, stroke, trauma, idiopathic cause, radiation, infections (mononucleosis) or multiple cranial neuropathy. Tumors were responsible for nearly half of the cases in different studies. We studied a female with hypoglossal nerve acute palsy. We made a differential diagnostic with others causes and a review of the literature. PMID- 16475545 TI - [Laryngeal amyloidosis. Bibliographic revision and actualisation]. AB - The amyloidosis is a uncommon and rare entity. It is a condition characterized by the extracellular deposition of homogeneous, eosinophilic proteinaceous material in the form of fibrils called "amyloid". This situation make a disfunction of the homeostasy in the affected place. In this letter we present three cases of localized laryngeal amyloidosis diagnosed between 1990 and 2004. Diagnosis and treatment were made in the three cases by direct laringoscopic and conventional microsurgery. Also we expose a actualised bibliographic review about studies more recent. PMID- 16475546 TI - [Otalgia as presentation of primary nasopharynx tuberculosis]. AB - We report a case of a 63-year-old woman who presented with left otalgia during more than six months. On examination, otoscopy was normal, whereas flexible nasopharyngoscopy revealed an asymmetry in the nasopharynx. CT scans confirmed the presence of a left nasopharyngeal mass. A transnasal endoscopic removal of the nasopharyngeal lesion was performed. Necrotizing granulomatosis was found. Tuberculosis of the nasopharynx was considered the final diagnosis, but oral treatment was not started. After 36 months of follow-up, nasal endoscopy and CT images show no sign of recurrence. Discussion is focused on the inclusion of nasopharyngeal diseases in otalgia differential diagnosis, histopathological diagnosis of granulomas and the need of any further treatment for these limited cases of extrapulmonary tuberculosis. PMID- 16475547 TI - [Seasonal influence on the microbiological colonization on surfaces of milk liners and flushing adapters in a newly installed milking parlor]. AB - A field trial was conducted to evaluate long term and seasonal effects on the development of mictobiological colonization in a newly installed milking parlor. The milking equipment was a 2 x 34 side-by-side milking parlor in a commercial dairy herd in Germany. The 1st batch of samples was taken before beginning of milking and in weekly intervals for a period of two months in winter 2002. The 2nd batch of samples was collected in summer 2003. After sampling the diagnostic material was kept in Amies medium at 4 degrees C until analysis. Samples were analyzed from the mouthpiece, the shank of the liner and the flushing adapter. The samples were collected immediately after disinfection of the milking equipment. They were analyzed qualitatively and quantitatively according to the official German guidelines (Amtliche Sammlung von Untersuchungsverfahren, paragraph 35, LMBG). Mesophil aerobe total plate count, S. aureus, E. coli, Coliforms, Streptococci, Yeast and Lactobacilli were considered. Results indicate a relationship of microbiological colonization of the surface of the milking technique with season and location of sampling. The microbiological colonization was higher in summer than in winter. There were more mesophil aerobe pathogens on the flushing adapters than on mouthpiece and shank of the liner. PMID- 16475548 TI - Inheritance of canine hip dysplasia: review of estimation methods and of heritability estimates and prospects on further developments. AB - Canine hip dysplasia is a long-known, widespread degenerative skeletal disease. A hereditary component of hip dysplasia was assumed early, although attempts to explain hip dysplasia with known Mendelian modes of inheritance did not sufficiently fit the data observed. Nevertheless, both recessive and dominant modes of inheritance were proposed. Later on, it was proposed that CHD was determined in a multifactorial way. Both the influence of many genes and environmental effects were assumed to affect the development of CHD. More recently, this thesis was supplemented and refined, as a major gene was detected as a cause of CHD in addition to a polygenic component. Nowadays, projects are under way with the aim to locate quantitative trait loci (QTL) significantly linked to CHD, and ultimately to develop gene tests to identify carriers of genes responsible for CHD. PMID- 16475549 TI - Oral vaccination against swine erysipelas--field experiences in Croatia. AB - In order to prove the effects of mass application of oral erysipelas vaccine via drinking water, in a farrow-to-finish production unit in Croatia, the growing finishing animals were divided into 3 groups and treated as follows:--Group 1 (n=199) was vaccinated intramuscularly against swine erysipelas at 1 week and 3 weeks after arrival in the growing-finishing facility with a swine erysipelas bacterin.--Group 2 (n=199) were vaccinated at the same time with an avirulent culture of Erysipelothrix rhusiopathiae oral vaccine through drinking water.- Group 3 (n=200) was not vaccinated. Animals with clinical signs of swine erysipelas, chronic progressive arthritis at slaughter, mortality, average daily weight gain during the growing-finishing phase were evaluated. None of the pigs in the groups 1 and 2 showed clinical signs typical for acute swine erysipelas. Twenty-four of the pigs (12 %) in group 3 had pyrexia and skin lesions typical for swine erysipelas. Fifteen pigs in group 1, 13 pigs in group 2, and 63 pigs in group 3 had chronic progressive arthritis (group 1 and 2 vs. group 3: P < 0.01). No significant differences in mortality were recorded between the groups. Group 1 and 2 had higher (P < 0.05) average daily weight gains compared with the group 3. PMID- 16475550 TI - [What did pigs look like in the Middle Ages?]. AB - The conformation of medieval swine is less well documented by written reports or osteological states. This paper evaluates the conformation of medieval pigs from about 140 pictures of various sources (books of hours, misericords, illustration of bibles text or proverbs, sculptures etc.). The pictures demonstrate that medieval pigs mainly were long legged, slender, razor backed, bristly with long snouts and prick ears (Tab. 1) well adapted for herding in forests and waste land. In mediterranean countries in early and high Middle Ages as well as in some regions of France and England in the late Middle Ages compact animals with shorts snouts and lopears were depicted. Probably these typs resulted from other housing and feeding conditions (stall feeding with wastes from fields, gardens, kitchen etc.). Medieval pigs were one-coloured white, grey, black and sometimes red, pictures from saddle-back animals came from Italy or south Germany. PMID- 16475551 TI - [Determination of the elastic properties of the compact bone in the femur of dogs]. AB - Total hip endoprotheses are a good possibility for treatment degenerative wear and pathologic damage of the hip joint in man as well as in dogs. However, aseptic loosening of the protheses, especially in the area of the shaft, is still a problem in conventional total hip endoprotheses. The purpose of the present study was to use the finite-element-analysis (FEA), to enhance endoprothetic design and to prevent loosening of protheses. In order to simulate the femur of the dog for numerical analysis, a material law for the compacta in the femur of the dog was developed. The elastic properties of the compacta were experimentally determined by using compression tests of bone samples of euthanised dogs. The results show constant denseness and a constant axial elastic modul in the compacta. PMID- 16475552 TI - [Intracranial fat bodies and their potential effect on brain composition and behaviour in domestic ducks with feather crests (3 case studies)]. AB - Intracranial fat tissue was found in the brains of three crested ducks. The three ducks differed in the size of their crest and in the volume and the location of the fat body within their brains. The duck with the large crest showed a fat body which counts for 19 % of its brain volume. Due to this fat accumulation, brain structures, mainly the cerebellum, were moved laterally. This duck had serious problems in motor coordination. Fat body of the second duck with a middle sized crest was situated in neostriatum and constituted 0,6 % of total brain volume. Additionally this duck displayed an encephalocele. The last duck did show a small crest. Its fat body was found in the area of the tentorium cerebelli and made up 17 % of its brain volume. The later two ducks were not hampered behaviourally. PMID- 16475554 TI - Patient navigation: new weapon in the DM arsenal? PMID- 16475553 TI - [Case report: mast cell leukosis in a neonatal calf]. AB - Multicentric mast cell tumours in a newborn Fleckvieh-calf are described. The calf showed clearly pronounced lesions over the whole body. The lesions were multiple raised, cutaneous, greyisch-red and partially ulcerated. It died three hours after birth. Pathohistological examinations resulted in multiple mast cell tumours within the dermis. In addition multifocal to diffuse mast cell aggregations were observed in several internal organs including the lymph nodes and the bone marrow. No evidence for the presence of bovine leukemia virus was found by both investigating a lymph node homogenate of the calf and a blood sample of the mother cow. In this paper the pathomorphology of this rare disease is described, a possible cause is discussed and a short review of the available literature is presented. PMID- 16475555 TI - Targeting asthma control in youngsters under the age of 12. AB - It's about time because despite the high incidence of asthma in youngsters, there has been no validated way for practitioners to assess the level of disease control in this group. Part of the problem is that parents routinely underestimate severity of symptoms, according to research. As a result, young patients often receive inadequate treatment. To resolve the problem, developers have come up with a seven-item screen that can deliver the information busy practitioners need before they even enter the exam room. PMID- 16475557 TI - DM program targets the chief health care decision-makers: women. AB - The program is targeted toward women over the age of 30. However, developers at Eli Lilly and Company believe their new DM offering, HealthInsights for Women, will offer benefits to spouses, children, and aging parents as well. The program currently addresses three highly prevalent conditions, but plans are already underway to expand the approach further. PMID- 16475556 TI - A call to action on health care disparities among minorities. AB - That is according to a cadre of health care leaders who contend that health care quality cannot improve until America successfully addresses glaring inequities that leave many racial and ethnic minority populations at a disadvantage. In an effort to dramatically shift thefocusfrom research and discussion of disparities to development of practical interventions, the Robert Wood Johnson Foundation is putting millions of dollars behind a series of initiatives designed to identify and disseminate real-world solutions to the problem over the next three to four years. PMID- 16475558 TI - A silver lining. PMID- 16475559 TI - 2006 coding update--Part I. PMID- 16475560 TI - Regulatory compliance associated with contrast media. AB - A basic understanding of the role of regulatory agencies in governing the healthcare environment and their influence over contrast media use is required of radiographers and imaging administrators to meet the many standards of compliance. In addition, radiology management teams must consider cost effectiveness, departmental efficiency, workplace safety, and compliance in choosing to implement new products. Regulatory agencies may be classified into 2 groups, voluntary and involuntary. Involuntary agencies are governmental agencies mandating regulatory compliance by local, state, or federal laws. Voluntary agencies are precisely that, those agencies an institution voluntarily chooses to participate with, to demonstrate the quality of care they provide. Failure to follow involuntary regulatory guidelines or to participate in voluntary best practice standards jeopardizes patient safety and the quality of care provided, and exposes the institution and the individual to liability risks. Severe penalties may result from a failure to maintain regulatory compliance, including the possibility of large fines, criminal indictments, and loss of third-party reimbursement. Achieving regulatory compliance is never an easy venture with the number of regulatory agencies and standards needing to be addressed. Combining regulatory compliance with the effects of doing business provides quite a challenge for today's imaging departments. A solid knowledge base in regulatory standards along with continuous investigation of new standards will allow departments to evaluate their own processes involved in providing patient care. Recognition of areas of high risk/high volume, including contrast media use, will assist in directing the departments' focus appropriately. A thorough evaluation of the products used and their respective handing and administration, in regard to patient and workplace safety, and appropriate documentation of workplace injuries due to contrast media packaging, will assist in maintaining a high level of compliance. PMID- 16475561 TI - CR/DR systems: what each technology offers today; what is expected for the future. AB - Computed radiography (CR) is considered by some to be the work-horse for digital image capture of general radiography exams because it is affordable, offers excellent image quality and exposure latitude and utilizes existing x-ray systems. CR systems deliver digital imaging to general radiology departments and lower-volume areas that can include hospital floors and outpatient imaging centers. Digital radiography (DR) technology is more expensive, but some believe it earns its keep with significant productivity gains and the capacity for higher image quality or lower dose. DR systems are especially appropriate for emergency room settings and high-volume areas in general radiology departments, orthopedic clinics, imaging centers and other facilities. Facilities with growing patient volumes and limited space often choose to install DR systems in one or 2 exam rooms to double the productivity of those rooms, while one or more CR systems serve the remaining rooms or remote areas. Patients benefit from both faster image capture (it takes less time for each imaging exam) and hospitals achieve a digital distribution process that speeds delivery of radiology reports to referring physicians and a more efficient imaging workflow that can lead to increased revenues. PMID- 16475563 TI - Building the bubble. PMID- 16475562 TI - Maximizing the imaging center revenue cycle: patient throughput. PMID- 16475565 TI - PACS--the gift that keeps on giving. PMID- 16475564 TI - Recruiting healthcare professionals to rural areas. AB - Maintaining an adequate number of healthcare providers for the nation's most underserved populations is increasingly diffiicult. Rural medical services have issues that often complicate recruitment and retention of qualified medical professionals. This review of literature examines some of the issues unique to rural areas. Consideration of these issues during recruitment strategies may lead to increased recruitment and retention of healthcare professionals to ruraI areas. PMID- 16475566 TI - Who will mind the store? PMID- 16475567 TI - A special kind of person. PMID- 16475568 TI - Nurses warn Alzheimer's drugs ban will lead to more sedation. PMID- 16475569 TI - Nurses may be forced to bear the brunt of the AfC funding shortfall. PMID- 16475570 TI - Give older people enough time to fully recover. PMID- 16475571 TI - Staying within the law. PMID- 16475572 TI - 'Clients respect and appreciate us'. PMID- 16475573 TI - Increasing the awareness of patient misidentification. AB - The scale of misidentification of patients in the health service is significant and can lead to incorrect treatment and even death. The National Patient Safety Agency has recently issued advice on how to prevent misidentification in hospitals. Within the health service as a whole, there is still potential for improved practice. PMID- 16475574 TI - Detached retina. PMID- 16475575 TI - Assessment of consciousness. Part two. PMID- 16475576 TI - Principles of early intervention in the treatment of psychosis. AB - Early intervention in psychosis is the first of a new wave of preventive strategies in psychiatry. Evidence from well-conducted studies suggests that it can prevent the development of the long-term debilitating negative symptoms of schizophrenia. This article discusses the research basis of the interventions, practical issues around its implementation and some possible limitations. PMID- 16475577 TI - Lactulose. PMID- 16475578 TI - Facilitating collaboration using a clinical teams programme. AB - The Clinical Teams Programme aims to develop clinical leadership. This article describes how it was used to improve teamworking at Reading PCT. The programme enhanced clinical leadership by improving confidence, allowing the development of effective teams and providing solutions to improve existing practice and patient care. PMID- 16475579 TI - Provision of palliative care education in nursing homes. AB - AIM: To map the nature and extent of existing palliative care education activities. METHOD: Data was gathered from questionnaires, face-to-face and telephone interviews, visiting palliative care teams across Mount Vernon Cancer Network and attendance at conferences, meetings and seminars. A comprehensive needs assessment for palliative care education within nursing homes was completed. RESULTS: The findings revealed inequality across the network with regard to education provision and uptake of palliative care services. Recruitment of overseas staff and a transient workforce were both cited as major difficulties in implementing education programmes. Funding of these programmes and responsibility for providing the education remain unclear. CONCLUSION: There was a real and urgent need for palliative care training in the network area and there was scope for a variety of approaches to be adopted to deliver the required training. PMID- 16475580 TI - How extending the hand of friendship builds better lives. PMID- 16475581 TI - Heart failure nurses and the voluntary sector. PMID- 16475582 TI - Young people and sexuality: consent and confidentiality. AB - Coping with consent and confidentiality issues relating to teenage sexual health can be challenging for nurses. Kathy French explains why it is vital that all those involved with young people have the knowledge and competence to deal with questions concerning consent for treatment, have regular training in child protection principles and are familiar with local policies. PMID- 16475583 TI - A practical guide to insulin pens. AB - Health professionals need to be conversant with the different types of pens and insulins available so that they are able to offer informed advice to patients. Pat Miles highlights the importance of their having good communication skills, as they have an important role in educating patients how to manage their diabetes. PMID- 16475584 TI - Rubella and the MMR vaccine. AB - Rubella is often thought of as a disease of childhood that has few complications. However, congenital rubella resulting from transmission from mother to baby during pregnancy can result in multiple severe defects. There are concerns about the uptake of the MMR vaccine and that any outbreak of rubella would affect disproportionately minority groups in the UK who may not have had the vaccine. Helen Bedford explains. PMID- 16475585 TI - Managing bleeding malignant skin lesions. AB - Malignant skin lesions can bleed as a result of the tumour itself or after the application of inappropriate dressings. Vivien McMurray discusses some of the measures that can be taken to control light bleeding, together with other methods that should be considered when profuse bleeding occurs. PMID- 16475586 TI - A stepping stone to full prescribing. PMID- 16475588 TI - Contemplating retirement: should I keep working? PMID- 16475587 TI - Acupuncture compared to the other alternative therapies. PMID- 16475589 TI - Concierge care and administrative fees. PMID- 16475590 TI - Protecting patients from the storm: TMA's Medical Liability Reform Campaign. PMID- 16475591 TI - Physician practices and safer medical devices. PMID- 16475592 TI - Loss prevention case of the month. Surgeon's friendship averts litigation? PMID- 16475593 TI - Medicare Health Support helps doctors care for chronically ill patients. PMID- 16475594 TI - It's not too late: reducing 2005 taxes through retirement plan contributions. PMID- 16475595 TI - A survey of Spanish language services among L&D units in Southeast Tennessee and North Georgia. AB - Over the past few years, we have noticed in our obstetrics residency program at Erlanger Hospital in Chattanooga that Hispanic women delivered by our service have become the majority of patients in our care. While the Hispanic population in Tennessee made up only 2.2 percent of the total state population in 2000, certain regions have seen large increases in immigrant population and Latina women accounted for 30 percent of our deliveries at the Baroness Erlanger Campus in 2003. Specifically, the number of women who speak little-to-no English has risen dramatically, forcing the hospital and all employees to expand efforts to communicate with patients in Spanish. PMID- 16475596 TI - Tennessee Shigella update. PMID- 16475597 TI - Psychiatric expert testimony. PMID- 16475598 TI - History of Tennessee Medicine, Part I. The old days. PMID- 16475599 TI - Medical Liability Reform and TMA Alliance. PMID- 16475600 TI - Regeneration of class II furcation defects: determinants of increased success. AB - One of the most important indications for guided tissue regeneration (GTR) treatment is class II furcation lesion. However, periodontal regeneration of this type of defect, although possible, is not considered totally predictable, especially in terms of complete bone fill. Many factors may account for variability in the response to regenerative therapy in class II furcation. The purpose of this review is to assess the prognostic significance of factors related to the patient (smoking, stress, diabetes mellitus, acquired immunodeficiency syndrome and other acute and debilitating diseases, and the presence of multiple deep periodontal pockets), local factors (furcal anatomy, defect morphology, thickness of gingival tissue and tooth mobility), surgical treatment (infection control, bone replacement grafts combined with barriers or GTR alone, type of barrier and surgical technique), and postoperative period (plaque control, membrane exposure, membrane retrieval and a regular supportive periodontal care program) for successful of GTR in class II furcations. PMID- 16475601 TI - Microhardness of resin-based materials polymerized with LED and halogen curing units. AB - The purpose of this study was to evaluate the microhardness of resin-based materials polymerized with a LED (light-emitting diode) light-curing unit (LCU) and a halogen LCU. Twenty cylindrical specimens (3.0 mm in diameter and 2.0 mm high) were prepared for each tested material (Z100, Definite and Dyract). Specimens were light-cured with two LCUs (Ultraled and Curing Light 2500) for either 40 or 60 s on their top surfaces. Hardness was measured on top and bottom surfaces of each specimen. Statistical analysis was done by ANOVA and Tukey's test (p<0.05). There was no significant difference in hardness between LED LCU and halogen LCU for Z100 and Dyract on top surface. Conversely, lower hardness was recorded when Definite was light-cured with the LED LCU than with the halogen lamp. On bottom surface, hardness was significantly lower for all materials light cured with LED LCU. Z100 was harder than Dyract and Definite regardless of the light curing unit. There was no significant difference in hardness between the exposure times on top surface. Higher hardness was obtained when the materials were light-cured for 60 s on bottom surface. The tested LED was not able to produce the same microhardness of resin-based materials as the halogen LCU. PMID- 16475602 TI - Effect of a carbamide peroxide bleaching gel containing calcium or fluoride on human enamel surface microhardness. AB - This in vitro study evaluated the surface microhardness of human enamel submitted to bleaching with 10% carbamide peroxide (CP) containing calcium or fluoride. Ninety-eight dental blocks (5 x 5 mm2) with polished enamel surfaces were randomly assigned to 7 treatment groups (n=14), as follows: without bleaching and storage in artificial saliva (control); 10% CP; 10% CP + 0.05% calcium; 10% CP + 0.1% calcium; 10% CP + 0.2% calcium; 10% CP + 0.2% fluoride; and 10% CP + 0.5% fluoride. During 14 days, enamel surfaces were daily exposed to a 6-h bleaching regimen followed by storage in artificial saliva. Surface microhardness was measured before (baseline), during (7th day), immediately after bleaching (14th day) and 1 week post bleaching. Data were analyzed by two-way ANOVA and Tukey's test (p<0.05). All treatments reduced SM significantly during the bleaching cycle (7th day), immediately after bleaching (14th day) and 1 week post bleaching, compared to baseline and to the unbleached control group. In conclusion, in spite of the addition of calcium and fluoride, all bleaching treatments affected the enamel surface microhardness. PMID- 16475603 TI - Influence of pH of different adhesive systems on the polymerization of a chemically cured composite resin. AB - The purpose of this study was to investigate the effect of pH of different adhesive systems on the polymerization of a chemically cured composite resin (Adaptic--AD), by means of tensile bond strength testing. The adhesive systems tested were: ARM, Prime & Bond 2.1 (PB), Scotchbond Multi Purpose (SMP) and Single Bond (SB). Bond strength at the resin/adhesive system/resin interface was assessed. Five groups (n=5) were formed, according to following configuration: G1: AD/ARM/AD; G2: AD/PB/AD; G3: AD/SMP/AD; G4: AD/SB/AD; G5: AD/AD (no adhesive). A two-mold stainless steel matrix with a cone-shaped opening (1-mm thick; 4 mm in diameter) was used to obtain resin discs. AD resin was inserted into the first mold, left-self curing and an adhesive layer was applied onto resin surface and light-cured. The second mold was assembled over the first and was filled with the resin. After 10 min, this setting was loaded in tension in a universal testing machine running at a crosshead speed of 0.5 mm/min. Data were submitted to one-way ANOVA and Tukey's test (p<0.05). Bond strength means (kgf) were: G1: 15.23 +/- 4.1; G2: 0.00 +/- 0.0; G3: 16.96 +/- 2.4; G4: 10.08 +/- 2.7; G5: 15.44 +/- 0.9. There were statistically significant differences (p<0.05) between G2-G1; G2-G3; G2-G4; G4-G1; G4-G3. The systems with the lowest pHs (PB and SB) yielded the lowest bond strength. The findings of this in vitro study demostrates that the pH of adhesive systems influences the polymerization and bond strength of chemically cured resin materials. The low pH simplified adhesive systems showed distinct degrees of incompatibility with the chemically cured resin, when compared to the conventional adhesive systems. PMID- 16475604 TI - Visual evaluation of in vitro cariostatic effect of restorative materials associated with dentifrices. AB - This study evaluated in vitro the cariostatic effect of 6 restorative materials with and without fluoride release (Fuji II LC, F-2000, Degufil Mineral, Sure Fil and Z-250) associated with a fluoridated and a non-fluoridated dentifrice (Sensodyne Original Formula and Sensodyne Sodium Bicarbonate), on human enamel. Class V-like cavities were prepared on 240 enamel slabs, assigned to 12 groups (6 materials and 2 dentifrices). After cavity restoration, the slabs were submitted to a thermocycling regimen of 1000 thermal cycles and demineralization/remineralization cycles. During pH cycles, slurries of fluoridated and non-fluoridated dentifrices were applied for 5 min. Formation of artificial caries-like lesions was scored independently and blindly by 5 calibrated examiners according to an ordinal scale ranked 0 to 3 by visual examination. The results were analyzed statistically by the Kruskal-Wallis test and pair-wise comparisons (alpha=0.05). There were no significant differences (p>0.05) among the restorative materials associated with the fluoridated dentifrice. When used in association with the non-fluoridated dentifrice, Ketac Fil showed the highest cariostatic effect followed by Fuji II LC and the other materials. Ketac-Fil was the only material that did not differ statistically when combined with either the fluoridated or the non-fluoridated dentifrice. In conclusion, under the tested experimental conditions, the association of restorative materials and fluoridated dentifrice yielded higher cariostatic effect, except for the conventional glass ionomer cement, whose cariostatic effect was not influenced by the type of dentifrice. PMID- 16475605 TI - Effect of microwave treatments on dimensional accuracy of maxillary acrylic resin denture base. AB - Microwave energy has been used as an alternative method for disinfection and sterilization of dental prostheses. This study evaluated the influence of microwave treatment on dimensional accuracy along the posterior palatal border of maxillary acrylic resin denture bases processed by water-bath curing. Thirty maxillary acrylic bases (3-mm-thick) were made on cast models with Classico acrylic resin using routine technique. After polymerization and cooling, the sets were deflasked and the bases were stored in water for 30 days. Thereafter, the specimens were assigned to 3 groups (n=10), as follows: group I (control) was not submitted to any disinfection cycle; group II was submitted to microwave disinfection for 3 min at 500 W; and in group III microwaving was done for 10 min at 604 W. The acrylic bases were fixed on their respective casts with instant adhesive (Super Bonder) and the base/cast sets were sectioned transversally in the posterior palatal zone. The existence of gaps between the casts and acrylic bases was assessed using a profile projector at 5 points. No statistically significant differences were observed between the control group and group II. However, group III differed statistically from the others (p<0.05). Treatment in microwave oven at 604 W for 10 min produced the greatest discrepancies in the adaptation of maxillary acrylic resin denture bases to the stone casts. PMID- 16475606 TI - Assessment of flexural strength and color alteration of heat-polymerized acrylic resins after simulated use of denture cleansers. AB - The purpose of this study was to assess flexural strength and color alteration of acrylic resins immersed in denture cleansers for different periods of time. Rectangular specimens (65 x 10 x 3mm) made from three heat-polymerized acrylic resins (Lucitone 550, QC-20 and Triplex) were assigned to three denture cleansers groups (Bony Plus, Corega Tabs and Efferdent Plus) and a control group (immersion in water). Soaking trials of 15 min and 8 h simulated 30 days of use. Flexural strength testing was carried out with 105 specimens on a universal testing machine. Color alterations were visually assessed by examination of photographs taken from 21 specimens. Flexural strength means (in MPa) were analyzed statistically by analysis of variance and Tukey's test at 5% significance level. There were significant differences (p<0.01) among the resins Lucitone (89.439 +/- 7.962), Triplex (88.024 +/- 5.167) and QC-20 (83.379 +/- 7.153). No significant differences (p>0.05) were found either among the denture cleansers (Bony Plus = 87.693 +/- 6.943; Corega Tabs = 86.955 +/- 7.114; Efferdent Plus = 86.195 +/- 7.865 and control = 86.536 +/- 7.012) or between the soaking periods (15 min = 86.875 +/- 7.625 and 8 h = 87.432 +/- 7.355) throughout the soaking cycles simulating 30 days of use. No color alterations were identified by visual examination. The findings of this study showed that chemical denture cleansers used according to the manufacturers' specifications did not cause flexural strength alterations or color changes in heat-polymerized acrylic resins submitted to soaking cycles that simulated 30 days of use. PMID- 16475607 TI - Effect of denture surface glazing on denture plaque formation. AB - This study evaluated, in vivo, the efficacy of a denture glazing material (Palaseal) in modifying plaque colonization of dentures. Ten subjects were selected and received maxillary temporary partial removable dentures, with complete acrylic palatal coverage. The right half of the fitting surface of the denture bases were glazed with Palaseal, whereas the other half was not glazed. One month after insertion, two fragments of the resin base of all dentures were removed (one from the glazed side and another from the non-glazed side). These samples were prepared and examined by scanning electron microscopy. Three months after insertion, other fragments were obtained and analyzed. Microscopic observation at 1 month revealed that, for all patients, the plaque film was thinner on the treated side in comparison to the non-treated side. However, at the 3-month evaluation, some areas of the glaze showed cracking, and both glazed and non-glazed sides were covered by a dense bacterial plaque film. In conclusion, the findings of this clinical experiment showed that glazing denture's fitting surface did not prevent bacterial colonization, but favored plaque removal while the glaze layer remained intact. After three months, glaze cracks created microretentive areas that increased plaque accumulation. PMID- 16475608 TI - Modified functional impression technique for complete dentures. AB - This report describes the use of a removable acrylic resin tray handle that can be easily attached to custom impression trays to produce an improved peripheral sealing zone. This device can be indicated to develop functional impressions for complete dentures using the patient-conducted muscular motion technique. In upper trays, the handle is fixed in the midline with acrylic resin, while in lower trays the centrally positioned handle is removed before border molding. This removable handle allows patient's suction and free tongue movements. Final impression is carried out in two stages: peripheral sealing (low fusion compound) and recording of the main supporting region of the denture (zinc oxide and eugenol paste). All border records are obtained from the patient's own movements (handle suction and tongue motion). The removable handle is simple to use, is reusable, can be adapted to any individual acrylic resin trays and allows accurate registration of the peripheral sealing zone (border tissues). PMID- 16475609 TI - Clinical evaluation of interocclusal recording materials in bilateral free end cases. AB - Five combinations of materials commonly used for intermaxillary records in cases of bilateral free end saddle were evaluated: acrylic resin base + wax; acrylic resin base + wax + ZOE paste; acrylic resin base + wax + Duralay resin; condensation silicone; and acrylic resin plate + irreversible hydrocolloid. The materials were evaluated by measuring the vertical distance between pairs of reference points located on the bases of the upper and lower articulated casts. The measurements obtained by manual articulation of the casts were used as the standards to which the measurements obtained with the recording materials were compared. ANOVA revealed significant differences among the materials and the Tukey's test showed that condensation silicone differed significantly (p<0.05) from the acrylic resin + wax combination, while no significant differences (p>0.05) were observed among the remaining materials. The condensation silicone presented the greatest differences from the measurements obtained by manual articulation of the casts whereas the acrylic resin base + wax combination presented the least differences. Based on the findings of this study, the following scale of fidelity can be settled from best to worst: acrylic resin base + wax; acrylic resin plate + alginate; acrylic resin base + wax + ZOE; acrylic resin base + wax + Duralay; and condensation silicone. PMID- 16475610 TI - Comparative study of the sealing ability of a polydimethylsiloxane-based root canal sealer. AB - The purpose of this study was to compare the sealing ability of three root canal sealers, RSA (polydimethylsiloxane sealer), Endion (glass-ionomer sealer) and Topseal (epoxy resin sealer), with and without smear layer removal. Ninety extracted human teeth were selected and assigned to 6 groups (n=15), according to the following protocols: in 3 groups, the smear layer was left intact and the root canals were obturated with gutta-percha points and RSA, Endion and Topseal, respectively; in the other 3 groups, the smear layer was removed and the root canals were obturated in the same way as described above. Microleakage was measured at 7 days, 1 month and 2 months, using the fluid transport model. The results were expressed in microL/24 h. Data were analyzed statistically by Kruskall-Wallis and Mann-Whitney non-parametric tests. The results showed that the Topseal group without smear layer leaked significantly less (p<0.05) than the RSA group without smear layer, at all experimental periods. No statistically significant differences (p>0.05) were found among the other groups, at the established evaluation intervals. The findings of this study showed that the polydimethylsiloxane sealer (RSA) did not yield better sealing ability than the other sealers, either in the presence or in the absence of smear layer. Topseal had the least microleakage from the root canal sealers evaluated. PMID- 16475611 TI - Comparative study of MTA and other materials in retrofilling of pulpless dogs' teeth. AB - This in vivo study compared the effect of mineral trioxide aggregate (MTA), IRM, Super EBA and ZOE in a puttylike consistency, used as retrofilling materials, in the healing process of periapical tissue of pulpless dogs' teeth submitted to a conventional retrofilling technique. Twenty-four premolars obtained from three dogs were used. At the first intervention, the animals were anesthetized, coronal access was obtained and pulpectomy was done. Root canals were kept open to the oral environment for 180 days to induce the formation of apical lesions. After surgical removal of the lesions with curettes, 2 mm of the apical root was cut out perpendicular to the long axis of the teeth, and root-end cavities were shaped with a low-speed round bur. The bone cavities were irrigated and dried, and the root-end cavities were filled with MTA, IRM, Super EBA and ZOE in a puttylike consistency. The bone cavities were passively filled with blood and flaps were sutured. The coronal access openings were cleaned and double-sealed with ZOE and amalgam. After 180 days, the animals were killed by anesthetic overdose, maxilla and mandible were removed and the pieces were processed for histomorphologic analysis. Data were evaluated blindly on the basis of several histopathologic events and the scores obtained were analyzed statistically using the Kruskal Wallis test. No significant differences were observed among MTA, Super EBA and IRM (p>0.05). However, ZOE had a significantly more negative influence on the apical healing (p<0.05). In conclusion, MTA, Super EBA and IRM had similar histopathologic effects among each other and better performance than ZOE used in a puttylike consistency. Furthermore, only MTA stimulated hard tissue deposition in direct contact with the retrofilling material, even when it was inserted under critical conditions. PMID- 16475612 TI - Osteoblast differentiation of human bone marrow cells under continuous and discontinuous treatment with dexamethasone. AB - Dexamethasone (Dex) has been shown to induce osteoblast differentiation in several cell culture systems. This study investigated the effect of continuous and discontinuous treatment with Dex on osteoblast differentiation of human bone marrow stromal cells (BMSC). Primary culture and first passage were cultured in media with or without Dex 10(-7) M. During the culture period, cells were incubated at 37 degrees C in humidified atmosphere of 5% CO2 and 95% air. At 7, 14, and 21 days, cell proliferation, cell viability, total protein content, alkaline phosphatase (ALP) activity and bone-like formation were evaluated. Data were compared by two-way analysis of variance. Dex did not affect cell viability and total protein content, but reduced cell number. ALP activity and bone-like formation increased when only first passage or both primary culture and first passage were treated with Dex, in comparison to the groups that did not have contact with Dex after first passage. The results of this study indicate that, for human BMSC, continuous presence of Dex did not appear to be required for development of the osteoblast phenotype, but Dex must be present after first passage to allow osteoblast differentiation expressed by reduced cell proliferation and increased ALP activity and bone-like formation. PMID- 16475613 TI - TP53 mutations in salivary gland neoplasms. AB - Many studies have demonstrated that loss of TP53 gene function has an important role in the genesis of many neoplasms, including salivary gland neoplasms. The purpose of this study was to examine the mutation profile of the TP53 gene in salivary gland neoplasms. Genomic DNA was extracted from paraffin-embedded tissues of pleomorphic adenoma, carcinoma in pleomorphic adenoma, mucoepidermoid carcinoma, adenoid cystic carcinoma and polymorphous low grade adenocarcinoma. Exons 5 to 8 of the TP53 gene were amplified by polymerase chain reaction (PCR) to perform single-stranded conformational polymorphism (SSCP) analysis. Band shifting was observed in exons 5, 6 and 8 in 9 out of 18 neoplasms. The results of this study suggest that mutations in TP53 gene are related to salivary gland neoplasms pathogenesis and that exons 5 and 8 are most frequently involved. PMID- 16475615 TI - [A number of guidelines for managing hypertension, but none are used. How general practitioners evaluate hypertension]. PMID- 16475614 TI - Accidental displacement of impacted maxillary third molar: a case report. AB - An unusual case of an impacted right maxillary third molar that was accidentally displaced into the maxillary sinus during exodontia and was surgically retrieved almost 2 years later is described. The tooth was removed under general anesthesia, after maxillary sinus exposure through Caldwell-Luc approach. Postoperative recovery was uneventful. Six months after the retrieval surgery, the maxillary sinus was completely healed and the patient did not present any complaint. PMID- 16475616 TI - [Diagnosis of hypertension in general practice. Unnecessary procedures]. PMID- 16475617 TI - [Which criteria count in chronic wounds. "TIME" heals wounds]. PMID- 16475619 TI - [When vaccination takes place after the bite, every hour counts. Race with the rabies virus]. PMID- 16475618 TI - [Prolactinoma and acromegaly. What can medication do, when must the surgeon be consulted?]. PMID- 16475620 TI - [New therapy options in inflammatory dermatoses. Can "light vaccination" really help? (interview by Dr. Judith Neumaier)]. PMID- 16475621 TI - [Cancer due to mobile radiofrequency masts. Caution: this study had significant shortfalls]. PMID- 16475622 TI - [Every 3rd diagnosis can be established only after talking with the patient. Are you an attentive listener?]. PMID- 16475624 TI - [The doctor-patient conversation: profession and art]. AB - Communication errors between physician and patient are the most frequent cause of treatment errors. On the other hand, a structured dialogue expedites the diagnostic and therapeutic processes. It is based on understanding, perceiving emotions, clarifying the patient's wishes, expanding options, defining goals as well as the prompt preparation for possible setbacks. PMID- 16475623 TI - [Self-evaluation of the practicing physician's communication ability]. AB - The lack of communication between physicians and patients is a complaint originating from mostly patients. However, many physicians also feel that they are poorly trained in this area. In a questionnaire that was published in the MMW 13/2005, 171 physicians rated the quality of doctor-patient communication, evaluated their own communicative competence and provided information on primary preconditions and hindrances to an effective doctor-patient dialogue. Eighty-five percent of the questioned viewed the quality of communication as "very important" or "important" for the success of the treatment. In addition to insufficient training in conducting a dialogue even under difficult circumstances, 46% cited the shortage of time in the practice, while 29% cited the compensation system and 12% cited work organization as the greatest impediment to good listening skills. At least one-fourth experience two to three situations a day in which the conversation is cut short because of a lack of time. PMID- 16475625 TI - [Differential diagnosis of jaundice]. AB - Jaundice is a symptom with a multitude of possible causes. These can be divided up into primary diseases of bilirubin metabolism, secondary hyperbilirubinemia in patients with liver disease, and diseases with bile duct occlusion. The major objective of the examination must be to exclude those causes that represent an acute danger to the patient, in particular cholangitis or cholecystitis. Symptoms that should cause alarm bells to ring include abdominal pain, fever and chills. When obtaining the patient's anamnesis, particular attention must be paid to lithiasis and previous operations. Laboratory findings of elevated alkaline phosphatase and gamma-GT indicate the presence of cholestasis. If an extrahepatic obstruction is suspected, ultrasonography of the upper abdomen is required. PMID- 16475626 TI - [In persistent asthma always think of antiinflammatory agents. Inhaled corticoids as basic medication in guideline specified asthma therapy]. PMID- 16475627 TI - [Conflict with national health insurance over inciting DMP boycott escalates. General practitioners threatened with million euro legal suit and revocation of licensure]. PMID- 16475628 TI - [L-thyroxine-iodine combination. Effective therapy in thyroid diseases]. PMID- 16475629 TI - [Many illnesses, early death. The ailments of young Mozart]. PMID- 16475630 TI - [A young man with breasts]. PMID- 16475631 TI - [Fats on the hip]. PMID- 16475632 TI - [Fibromyalgia: understanding the disease and its social implications]. PMID- 16475633 TI - [Low-dose 0.25% spinal levobupivacaine with epidural extension for cesarean section: comparison with 0.5% hyperbaric bupivacaine]. AB - INTRODUCTION: When low doses of local anesthetic are used in combined spinal epidural anesthesia for cesarean section, an epidural catheter can be used to enhance a possibly incomplete block or insufficient dose. OBJECTIVE: To compare the efficacy of spinal 0.5% hyperbaric bupivacaine with fentanyl (20 microg) at a conventional high dose of 0.07 mg cm(-1) (group 1) vs 0.25% levobupivacaine at a low dose of 7.6 mg with fentanyl (20 microg) and epidural volume extension with 6 mL of saline through an epidural catheter (group 2). MATERIAL AND METHODS: A randomized clinical trial enrolling patients scheduled for elective cesarean section. RESULTS: Sixty-two patients were studied (31 in each group). Characteristics in the groups were comparable at baseline and the maximum level of sensory block achieved was sufficient for all but 1 patient in group 2 who required general anesthesia. The bupivacaine dose in group 1 ranged from 10.5 to 12 mg. The motor block and duration was less intense in group 2 (P<0.0001) and patients in that group could be transferred out of the postanesthetic care unit earlier. CONCLUSIONS: The use of low doses of levobupivacaine with an opiate in combination with volume extension through an epidural catheter in the context of combined spinal-epidural anesthesia is a safe, effective technique that may allow the doses and motor block to be reduced when hyperbaric levobupivacaine is administered, without adverse events for patients. PMID- 16475634 TI - [Relation between glucose concentrations in cerebrospinal fluid and sensory and motor block during spinal anesthesia with hyperbaric bupivacaine]. AB - OBJECTIVE: To study the relation between cerebrospinal fluid (CSF) glucose levels, the highest level of sensory block, and the duration of motor block after intrathecal injection of 2 mL of hyperbaric bupivacaine. To determine CSF glucose levels upon recovery from motor block. PATIENTS AND METHODS: A prospective study of 34 patients administered a spinal anesthetic in continuous infusion through a 22-gauge catheter. CSF samples were extracted through the catheter 5, 10, 15, 20, 30, 45, and 60 minutes after start of infusion and upon motor recovery. After each extraction the intensity of the motor block was assessed; the intensity of the sensory block was assessed after each extraction up to 20 minutes. RESULTS: Glucose concentrations in CSF tended to decrease from 5 minutes (1027.07 [SD 349.04] mg dL(-1)) until full motor recovery (247.50 [20.39] mg dL(-1)). The probability of finding a motor block at a CSF glucose concentration of 287.5 mg dL(-1) or higher was less than 5%. We identified a positive correlation between the highest level of sensory block and the duration of full motor block (r=0.62, P<0.01) and between CSF glucose levels at the moment of greatest sensory block and upon full motor recovery (r=0.50, P<0.01). CONCLUSIONS: After continuous spinal anesthesia with hyperbaric bupivacaine, glucose concentrations in CSF are directly related to the highest level of sensory block, the course of the blockade, and its reversal. PMID- 16475635 TI - [Risk factors for transfusion in primary knee arthroplasty]. AB - OBJECTIVE: Transfusion is becoming safer but is not free of risk. It is important to establish a good approach to transfusion management and calculate real losses. Risk factors for transfusion should be identified. MATERIAL AND METHODS: This was a prospective study of 102 patients who did not receive intraoperative autotransfusion of shed blood, selected from a group of 127 who were undergoing primary knee arthroplasty. We initially calculated the amount of blood shed. Then, by multivariate logistic regression analysis we identified the model that best predicted that a patient would require transfusion. Receiver operating characteristic curves were constructed and the area under the curves calculated. RESULTS: Mean (SD) blood loss was calculated to be 1786 (710) mL. The best model considered initial hemoglobin (Hb), weight, height, and sex as predictive factors: Probability = 1/ (1+e(-Z)), where Z = 11.542 - 1.074 x initial Hb (g/dL) - 0.039 x Weight (kg) + 0.031 x Height (cm) + 0.267 x (sex: male=1 or female=0). The area under the ROC curve was 0.805 (0.44). CONCLUSION: Initial Hb, which can be modified before surgery, is one of the factors that most affects whether or not the patient will need a transfusion. Therefore, one of our first objectives in the process of managing transfusion is to improve preoperative Hb values. PMID- 16475636 TI - [Usefulness of transesophageal echocardiography during myocardial revascularization surgery without extracorporeal circulation]. AB - OBJECTIVE: Intraoperative transesophageal echocardiography can be a highly useful monitoring technique during myocardial revascularization surgery when extracorporeal circulation (ECC) is not being used. Transesophageal echocardiography provides real-time images on both volume status and segmental myocardial contractility without interfering with the surgical field. PATIENTS AND METHODS: A total of 25 patients undergoing myocardial revascularization by sternotomy without ECC were monitored by transesophageal echocardiography during surgery. RESULTS: The 18 men and 7 women studied had a mean (SD) age of 71.3 (8) years. A third of them had hypertension and diabetes, 3 had suffered a cerebrovascular accident, and 2 had renal failure. Nine patients had a history of acute myocardial infarction and 3 had undergone angioplasty. Baseline echocardiograms on all patients established that 6 had a low ejection fraction (<30%). Twelve had altered segmental contractility, which was transient in 11 cases. Six patients had improved ejection fraction at the final assessment. Transesophageal electrocardiography also monitored volume status and the effects of inotropic drugs and beta-blockers in 83% of the patients. CONCLUSION: Transesophageal electrocardiography is a minimally invasive, safe, and precise way to directly monitor the beating heart in real time during myocardial revascularization without ECC. Image quality is good. PMID- 16475637 TI - [Jehovah's Witnesses refusal of blood: religious, legal and ethical aspects and considerations for anesthetic management]. AB - The refusal of Jehovah's Witnesses to agree to blood or blood product transfusion based on religious beliefs is one of the most challenging conflictive issues health care givers have to face today. Such conflict is a by product of the ideological and religious diversity in society today. The perioperative care of such patients constitutes a genuine challenge for anesthesiologists and surgeons from technical, scientific, ethical, and legal perspectives. We review the reasons why Jehovah's Witnesses refuse transfusion and discuss the ethical, legal, and anesthetic aspects of their care. The literature up to August 2005 was reviewed by MEDLINE search. The following search terms were used: Jehovah's Witnesses, anesthesia (and anaesthesia), legislation and jurisprudence, ethics, blood transfusion, alternatives, anemia (and anaemia), erythropoietin, trigger, and critical care. To further cover ethical and legal aspects, we reviewed current laws in Spain and similar practice settings. PMID- 16475639 TI - [Intraoperative high frequency oscillatory ventilation in 2 children undergoing lung surgery]. AB - Over the past 10 years, the application of high frequency oscillatory ventilation (HFOV) has been extended beyond the neonatal period. The technique is now used in various respiratory disease settings when conventional mechanical ventilation fails. Even though HFOV has become increasingly routine in some pediatric intensive care units, familiarity with it is still limited among anesthesiologists and surgeons and it is not often applied during surgery. We report our experience using HFOV during thoracic surgery on 2 pediatric patients, one aged 5 years and the other aged 1 month. The respective surgical procedures were to close a bronchopleural fistula and to obtain a lung biopsy in order to provide guidance for limiting therapeutic intervention. In both cases the procedure was performed without adverse effects and allowed medical interventions to be carried out. We conclude that it is possible to perform thoracic surgery in pediatric patients undergoing HFOV. This ventilation mode can be useful during surgery and teams that care for critically ill children should be familiar with the equipment. PMID- 16475640 TI - [Cerebrospinal fluid drainage in endovascular repair of thoracic aortic lesions: preliminary report of experience with 5 patients]. AB - Endovascular repair of thoracic or thoracoabdominal aortic lesions as an alternative to open surgery, to avoid the high complication and mortality rates associated with the conventional approach, does not eliminate the risk of postoperative paraplegia. We report on a series of 5 patients with different thoracic aortic lesions who underwent endovascular stent-graft repair procedures. We describe measures to drain cerebrospinal fluid to prevent ischemic spinal cord injury. We also review the anesthetic management of patients undergoing this type of surgery. PMID- 16475638 TI - [Aortopulmonary fistula complicating aortic arch aneurysm]. AB - Aortopulmonary fistulas secondary to thoracic aneurysms are difficult to diagnose and few cases are reported in the literature. We describe the case of a patient first admitted to another hospital with a diagnosis of right upper lobe pneumonia. After the patient had been a week in hospital without improving, a computed tomography scan of the chest revealed a saccular aneurysm of the aortic arch measuring 9 cm at its widest diameter. Hemodynamics and respiratory signs deteriorated a few hours after transfer to our hospital. Physical examination revealed a systolic murmur and poor peripheral perfusion. The patient was intubated and inotropic support was provided. Upon insertion of a Swan-Ganz catheter, given suspicion of septic shock, progressive elevation of mixed venous oxygen saturation was observed. Measurement of venous oxygen saturation was 74% in the right atrium and 93% in the pulmonary artery. Left-right shunt was diagnosed and an emergency aortography revealed the aortopulmonary fistula. Emergency surgery was performed with the patient in deep hypothermia and cardiocirculatory arrest. There were no adverse events, and postoperative recovery and clinical course were good. PMID- 16475641 TI - [Opsoclonus-myoclonus: paraneoplastic syndrome associated with renal adenocarcinoma]. PMID- 16475643 TI - [Simultaneous use of 2 Arndt-type endobronchial blockers to collapse the lung in congenital tracheal bronchus]. PMID- 16475642 TI - [Eclampsia in a pregnant drug addict]. PMID- 16475644 TI - [Transdermal buprenorphine and silent acute coronary syndrome]. PMID- 16475645 TI - [Combined spinal-epidural block for knee arthroplasty in a patient with a transplanted heart]. PMID- 16475646 TI - [Unilateral compartment syndrome secondary to prolonged lithotomy positioning and peripheral vascular disease]. PMID- 16475647 TI - [Spontaneous rupture of a splenic artery aneurysm: a rare cause of hemoperitoneum]. PMID- 16475648 TI - [Perioperative management of a patient allergic to multiple nondepolarizing neuromuscular blockers during treatment for cervical fracture and dislocation]. PMID- 16475649 TI - [Reduced allogenic transfusion requirements through reinfusion of processed shed blood]. PMID- 16475650 TI - [GlideScope-assisted fiberoptic bronchoscope intubation of the trachea]. PMID- 16475651 TI - [History of materials: from the stone age to the age of plastics]. PMID- 16475652 TI - [Physicochemical aspects of biomaterials used in urology]. PMID- 16475653 TI - [Biomaterials and biocompatibility]. PMID- 16475654 TI - [Latex: an historical biomaterial that should no longer be used today]. PMID- 16475655 TI - [Biomaterials used in contact with the urinary tract for urine drainage: catheters and ureteric stents]. PMID- 16475656 TI - [Synthetic reinforcement and support biomaterials for the treatment of incontinence and prolapes: tape and mesh]. PMID- 16475657 TI - [Synthetic biomaterials and metals: application to urethral prostheses]. PMID- 16475658 TI - [Injectable synthetic biomaterials: filling agents for the treatment of incontinence and vesicoureteric reflux]. PMID- 16475659 TI - [Biomaterials: application to complex prostheses in urology]. PMID- 16475661 TI - [Cell therapy and tissue engineering in urology]. PMID- 16475660 TI - [Biological materials]. PMID- 16475663 TI - [Institutions and bodies concerned by biomaterials]. PMID- 16475662 TI - [CE marking for medical devices]. PMID- 16475664 TI - [Clinical trials of medical devices]. PMID- 16475665 TI - [The medical devices vigilance system]. PMID- 16475666 TI - Young adults' retrospective reports of parenting by mothers and fathers: associations with current relationship quality. AB - The authors examined retrospective reports of both mothers' and fathers' parenting and young adults' (N = 75) relationship quality. Multiple regression analyses showed that, as predicted, young adults' retrospective reports of the positive parenting they experienced as children were significantly related to the extent by which they currently viewed (a) others as accessible and responsive, (b) their relationships with others as meaningful and important, and (c) themselves as able to form healthy relationships. Although both mothers' and fathers' parenting related to the quality of current relationships with parents, only reports of fathers' parenting were related to the quality of current relationships with a romantic partner. Fathers' parenting was also related to views of the self as being able to form secure and close relationships. The authors address both methodological and substantive explanations for these results. PMID- 16475667 TI - Hot or not: do professors perceived as physically attractive receive higher student evaluations? AB - Previous research investigating the influence of perceived physical attractiveness on student evaluations of college professors has been limited to a handful of studies. In this study, the authors used naturally occurring data obtained from the publicly available Web site www.ratemyprofessors.com. The data suggested that professors perceived as attractive received higher student evaluations when compared with those of a nonattractive control group (matched for department and gender). Results were consistent across 4 separate universities. Professors perceived as attractive received student evaluations about 0.8 of a point higher on a 5-point scale. Exploratory analyses indicated benefits of perceived attractiveness for both male and female professors. Although this study has all the limitations of naturalistic research, it adds a study with ecological validity to the limited literature. PMID- 16475668 TI - Improved episodic integration through enactment: implications for aging. AB - Enactment may improve memory for verb phrases by facilitating episodic integration of object-action components into a unitized whole. It is unclear, however, whether the influence of enactment on episodic integration is related to or independent of the strength of the preexisting semantic relationship between components. To address this issue, the authors examined the influence of enactment on memory for lists of semantically related object-action phrases ("Put money in the wallet") and semantically unrelated phrases created by repairing these objects and actions to make phrases that were unusual but still were possible to perform ("String a thread through the wallet," "Put money in the napkin"). As such, phrases in the related and unrelated lists were matched for familiarity of the individual components and differed only in the associative strength of the object-action relationship. Although verbatim recall of unrelated lists was poorer under standard verbal encoding conditions, enactment succeeded in bringing performance to the level of related lists, indicating that enactment's influence on episodic integration was independent of the semantic relatedness of the object and action components. Analysis of partial recall errors (accurate recall of only one component) suggested that enactment benefited recall in the unrelated lists by improving memory for the action and reducing fragmentation of the association, providing further support for the unitization view. This pattern of results was replicated in normal older adults, a population that exhibits particular difficulty with episodic memory for unrelated associations. The cognitive mechanisms by which enactment may improve episodic integration in both younger and older adults are discussed. PMID- 16475669 TI - Directed forgetting of a single item. AB - In two experiments, we investigated directed forgetting of a single item. We presented participants with two phone numbers to remember and instructed half of the participants to forget the first phone number. The first number was either learned on a single trial (Experiment 1) or on three trials (Experiment 2). In Experiment 1, there was evidence of directed forgetting as a result of differential encoding; the forget instruction affected recall and recognition for both phone numbers. In Experiment 2, the evidence favored differential rehearsal because the forget instruction affected recall but not recognition. These results indicate that instructions to forget can affect memory of single items. PMID- 16475670 TI - Investigating retrospective influences on induction in rats' responding for 1% sucrose when food-pellet reinforcement is upcoming. AB - When rats lever press for 1% sucrose reinforcement in the first half of a 50-min session, response rates are higher when food-pellet reinforcement will be available in the second half than when 1% sucrose will be available. Results of past research have suggested that, under some conditions, this induction effect is prospective in nature (i.e., controlled by the conditions of reinforcement in the present session). However, that research did not rule out the possibility that, under other conditions, retrospective factors (i.e., the conditions of reinforcement in the previous session[s]) could contribute. In the present study, rats responded in two types of session, one in which 1% sucrose reinforcement was available in both halves of the session and one in which 1% sucrose and food pellet reinforcement were available in the first and second halves, respectively. Which type of session was in effect unsignaled and session type alternated every session (Experiment 1), every second session (Experiment 2), or after at least 20 consecutive sessions of one type (Experiment 3). Across experiments, the results indicated that it takes several sessions of one type for observable retrospective effects to occur, but those effects are short lived. These results allow the authors to identify the mechanisms that must underlie induction. The authors also discuss induction as an animal model of anticipation. PMID- 16475671 TI - Is induction produced by upcoming food-pellet reinforcement the outcome of an increase in overall activity or in operant responding? AB - Researchers have demonstrated that rats reliably increase their rates of pressing a lever for 1% liquid-sucrose reinforcement if they will soon have the opportunity to press a lever for food-pellet reinforcement. In the present experiments, the authors investigated if this increase in response rates occurred because the upcoming food pellets produced an increase in all behaviors (i.e., general arousal) or an increase in only the specific operant response (i.e., lever pressing). The results of Experiments 1 and 2 showed that the appearance of induction in rats' lever pressing for 1% sucrose reinforcement when food-pellet reinforcement was upcoming did not coincide with increases in the frequency of running in a wheel or making a nonreinforced nose-poke response. On the other hand, in Experiment 3, the authors found the appearance of induction coincided with increase nonreinforced lever presses on an adjacent lever. These results shed doubt on the idea that induction is a result of a general increase in all activity, and suggest instead that the increase in responding that occurs during induction is limited to the operant response. PMID- 16475672 TI - Experimental study on a large animal model of a new thermoablation technique. AB - BACKGROUND: A novel technique of thermoablation, using a microtube to deliver pulses of hot water vapour, was tested on a large animal model in order to evaluate its efficacy and potential adverse effects. MATERIALS AND METHODS: The medical device consisted of a microtube extension connected to a hydropneumatic pump. Pulses of pure water were injected though the microtube where they were heated and delivered as vapour into the target zone. The method was tested on the liver of 12 healthy pigs, either during open surgery or percutaneously under ultrasounds. RESULTS: The technique was efficient and well-tolerated by the animals. Large volumes of necrotic tissue were created in a significantly short time compared to concurrent thermoablative techniques. CONCLUSION: Anticipating human application, this experimental study demonstrated a safe and efficient innovative thermoablation technique. The first human applications have been successfully performed and will be reported soon. PMID- 16475673 TI - Cancer-related overexpression of the peripheral-type benzodiazepine receptor and cytostatic anticancer effects of Ginkgo biloba extract (EGb 761). AB - The peripheral-type benzodiazepine receptor (PBR) is an 18-kDa high affinity drug and cholesterol-binding protein that is involved in various cell functions, including cell proliferation and apoptosis. PBR was shown to be overexpressed in certain types of malignant human tumors and cancer cell lines, correlating with enhanced tumorigenicity and cell proliferation rates. The present study was conducted in order to further define the role of PBR in cancer and to extend our recent findings regarding the possible anticancer effects of the standardized Ginkgo biloba extract EGb 761. Treatment with EGb 761 decreased PBR mRNA levels and inhibited the proliferation of breast, glioma and hepatocarcinoma cell lines, further corroborating our previous contention that its mechanism of action is through the modification of PBR expression. In vivo treatment with Ginkgo biloba extract led to dose-dependent decreases in xenograft growth of both MDA-MB-231 breast cancer and U-87 glioma cell lines in nude mice, although the effects were not maintained after 50 days of treatment in the latter. The results obtained in MDA-MB-231 xenografts indicated pronounced inhibition of tumor growth, verified by MRI imaging. These results were obtained using a modified experimental protocol where the animals were treated with the extract before cell inoculation. Although an exact role for PBR in relation to the initiation and progression of various types of cancer remains to be defined, our results indicate that PBR overexpression in certain cancer cells is related to an aggressive phenotype. Since EGb 761 treatment opposes this aggressive phenotype by decreasing PBR overexpression, it could be useful in preventing or treating cancer invasiveness and metastasis. PMID- 16475674 TI - Effects of zoledronic acid on proteinase plasma levels in patients with bone metastases. AB - BACKGROUND: The effects of the bisphosphonate derivative zoledronic acid (ZA) on the circulating levels of matrix metalloproteinase-2 (MMP-2), matrix metallo proteinases-9 (MMP-9), cathepsin B (Cath B) and urokinase-type plasminogen activator (uPA) in patients with bone metastasis (BMTS) and the possible correlation with the symptomatic response induced by this drug in these patients were evaluated. PATIENTS AND METHODS: Proteinase levels were determined by enzyme linked immunosorbent assay (ELISA) in the plasma of 30 patients with painful bone metastases from breast or prostate cancer undergoing multiple treatment with ZA (4 mg i.v., every 4 weeks). Healthy subjects (HS) of both genders (12 female and 30 male) served as the control group. The symptomatic response to ZA was assessed by the visual analog scale score (VAS). RESULTS: The median MMP-2 and MMP-9 pretreatment levels were more elevated in BMTS as compared to HS (p < or = 0.0001). Conversely, uPA levels were lower in BMTS p = 0.0033; no significant difference was observed for Cath B. ZA administration was associated with a symptomatic response (VAS score < or =4) in 25/30patients (83.3%) (p < 0.0001). This phenomenon paralleled a decrease of Cath B and MMP-2 plasma concentrations from baseline values on week 12 (p = 0.05). A similar trend, although not statistically significant, was also noted for MMP-9 and uPA. However, no direct relationship was observed between the analgesic effect induced by ZA and changes in the circulating levels of these enzymes. CONCLUSION: These data show that ZA administration may provide relief from bone pain in patients with diffuse skeletal metastases and confirm a possible implication of cysteine proteinases and matrix metalloproteinases in bone metastasis formation, but not in the pathogenesis of metastatic bone pain. PMID- 16475675 TI - Conjugated linoleic acid (CLA) up-regulates the estrogen-regulated cancer suppressor gene, protein tyrosine phosphatase gamma (PTPgama), in human breast cells. AB - BACKGROUND: Conjugated linoleic acid (CLA), a naturally occurring compound found in ruminants products, has been shown to possess anticancer properties in vivo and in vitro. There are several CLA isomers in ruminant-produced foods, among which t10, c12-CLA and c9, t11-CLA are the most potent. Protein tyrosine phosphatase gamma (PTPgamma) has been implicated as a tumor suppressor gene in kidney and lung cancers. Our previous results indicated that estradiol-17beta (E2)-induced suppression of PTPgamma may play a role in mammary tumorigenesis. MATERIALS AND METHODS: The effects of t10, c12-CLA and c9, t11-CLA on PTPgamma mRNA expression in human breast epithelial cells and stromal cells, isolated from surgical specimens of mammoplasty and breast cancer patients, were detected and quantified by RT-PCR RESULTS: The PTPgamma mRNA expression was lower in cancer than in normal breast cells. Both t10, c12-CLA and c9, t11-CLA significantly (p < 0.05) increased the PTPgamma mRNA levels in primary cultured normal breast epithelial cells, normal breast stromal cells and breast cancer epithelial cells, but not in breast cancer stromal cells. t10, c12-CLA appeared to be the most active isomer in estrogen receptor a (ERalpha)-positive human breast cancer epithelial cells. CONCLUSION: The results indicate that dietary CLA might serve as a chemo-preventive and chemo-therapeutic agent in human breast cancers by up regulating the estrogen-regulated tumor suppressor gene, PTPgamma expression. PMID- 16475676 TI - Identification of a new breast cancer-related gene by restriction landmark genomic scanning. AB - BACKGROUND: Molecular genetic analyses have demonstrated the importance of the accumulation of genomic changes in the etiology of cancer and, additionally, have identified valuable genetic markers for certain cancers. Although several prognostic markers have already been identified for breast cancer, it is clear that others remain to be identified. MATERIALS AND METHODS: Fourteen breast cancer samples and non-cancerous counterparts were applied to restriction landmark genomic scanning (RLGS) and 6 breast cancer cell lines (MCF-7, MDA-MB 435, T-47D, MDA-MB-231, SK-BR-3 and BT-20) and 9 cancer tissue samples were applied to reverse transcnptionalpolymerase chain reaction (RT-PCR) for screening of novel genetic alterations. RESULTS: Two spots were identified on the RLGS profiles of cancerous tissue that differed from those of normal tissue. Nucleotide sequencing and homology search analysis showed that these spots represented the voltage-dependent calcium channel alpha1H subunit gene (CACNA1H gene) and a locus immediately downstream of the growth factor receptor-binding protein 7 (GRB7) gene. Expression of the CACNA1H gene was confirmed by RT-PCR. CONCLUSION: Two genes, Grb7 and CACNA1H, were identified by RLGS. The expression of CACNA1H in breast cancer was confirmed for the first time. PMID- 16475677 TI - Antitumor effect of cordycepin (3'-deoxyadenosine) on mouse melanoma and lung carcinoma cells involves adenosine A3 receptor stimulation. AB - An attempt was made to elucidate the molecular targetfor the antitumor effects of cordycepin (3'-deoxyadenosine) using non-selective and selective adenosine A1, A2a, A2b and A3 receptor agonists and antagonists. Although adenosine and 2' deoxyadenosine (up to 100 microM) had no effect, cordycepin showed remarkable inhibitory effects on the growth curves of B16-BL6 mouse melanoma (IC50= 39 microM) and mouse Lewis lung carcinoma (IC50 = 48 microM) cell lines in vitro. Among the adenosine receptor agonists and antagonists used (up to 100 microM), only 2-chloro-N6-(3-iodobenzyl)-adenosine-5'-N-methyluronamide (Cl-IB-MECA), a selective adenosine A3 receptor agonist, notably inhibited the growth of both mouse tumor cell lines (B16-BL6; IC50 = 5 microM, LLC; 14 microM). In addition, the tumor growth inhibitory effect of cordycepin was antagonized by 3-ethyl 5 benzyl 2-methyl-6-phenyl-4-phenylethynyl-1,4-(+/-)-dihydropyridine-3,5 dicarboxylate (MRS1191), a selective adenosine A3 receptor antagonist. These results suggest that cordycepin exerts inhibitory effects on the growth of mouse melanoma and lung carcinoma cells by stimulating adenosine A3 receptors on tumor cells. PMID- 16475678 TI - DNA methylation of CHFR is not a predictor of the response to docetaxel and paclitaxel in advanced and recurrent gastric cancer. AB - BACKGROUND: Cell cycle checkpoint dysfunction is often associated with sensitivity to chemotherapeutic agents. In this study, the question of whether DNA methylation of CHFR, a mitotic checkpoint gene, can predict the response of advanced and recurrent gastric cancers (GCs) to docetaxel or paclitaxel was examined. MATERIALS AND METHODS: In a retrospective study, 41 patients with GC treated with paclitaxel alone (n=12) or a combination of docetaxel and S-1 (tegafur, 5-chloro-2,4-dihydroxypyridine, potassium oxonate) (n=29) were studied. The DNA methylation status of the CHFR gene was examined by combined bisulfite restriction analysis of DNAs from 41 GC tissues and the methylation status was compared to their sensitivity to chemotherapy. The levels of CHFR mRNA were measured by quantitative reverse transcription-PCR. RESULTS: DNA methylation of CHFR was found in 15 (36.6%) out of the 41 GC samples and the levels of CHFR mRNA were associated with the methylation status of CHFR (p = 0.034). In 41 samples of corresponding non-neoplastic mucosae, no DNA methylation of CHFR was detected. Among 12 patients treated with paclitaxel alone, only 1 (20.0%) of the 5 patients with CHFR methylation had a partial response (PR) to paclitaxel, whereas 3 (42.9%) of the 7 patients without CHFR methylation had a PR to paclitaxel (p = 0.836). In 29 patients treated with a combination of S-1 and docetaxel, there was no clear association between the CHFR methylation status and response to chemotherapy (p = 0.092). CONCLUSION: We conclude that the DNA methylation of CHFR alone cannot predict the response of advanced and recurrent GC to docetaxel or paclitaxel. Both paclitaxel and docetaxel may be effective for treatment of GC even if CHFR is expressed. PMID- 16475679 TI - Lymph node metastasis is infrequent in patients with highly-methylated colorectal cancer. AB - In this study, the combined methylation status of p16, p14, HLTF, CDH13 and RUNX3 was examined in 59 resected primary colorectal cancers using methylation-specific PCR and the methylation status was correlated with the clinicopathological features of the affected patients. A significant difference was found in lymph node metastasis (p = 0.0359) on comparing the methylation status in colorectal cancers with lymph node metastasis to those without. There was also a significant gender difference (p = 0.0248) when the methylation status was compared. Colorectal cancer without lymph node metastasis frequently exhibited aberrant methylation of the p16, p14, HLTF, CDH13 and RUNX3 genes. PMID- 16475680 TI - Tumour plasticity and extravascular circulation in ECV304 human bladder carcinoma cells. AB - BACKGROUND: The concepts of vasculogenic mimicry and mosaic vessels have been proposed as novel modes of tumour neovascularisation. However, the presence and significance of these types of neovascularisation remain unclear. MATERIALS AND METHODS: ECV304 human bladder carcinoma cells were used to determine how tumour cells take part in tumour neovascularisation. RESULTS: Subcutaneous ECV304 xenografts in mice showed various vessel types, including angiogenic vessels, tumour cell-related vessels and extracellular matrix networks. A tracer experiment demonstrated perfusion of beads in these structures. ECV304 cells, cultured on collagen I gels, formed tube networks with expressions of several endothelial-related markers. In coculture models of ECV304 cells and human umbilical vein endothelial cells, the two cells collaborated to form sprouts or networks. CONCLUSION: ECV304 cells possess an endothelial character which confers the ability to mimic and collaborate with vascular endothelial cells and facilitates the acquisition of tumour microcirculation. PMID- 16475681 TI - Immunohistochemical distribution of heat shock protein 47 (HSP47) in scirrhous carcinoma of the stomach. AB - BACKGROUND: The aim of the present study was to examine the immunohistochemical distribution of the 47-kDa heat shock protein (HSP47) to enhance the understanding of the mechanisms involved in stromal fibrosis, which accompanies cancer infiltration in scirrhous carcinoma of the stomach. MATERIALS AND METHODS: In vitro gastric cancer models were prepared by collagen gel cultures using three different human gastric cancer cell lines (KATO-III, MKN-74, MKN-45) and a human fibroblast cell line (TIG-101). Tumor tissues were obtained from ten patients with early gastric cancer (5 scirrhous carcinoma; 5 non-scirrhous carcinoma) and three patients with advanced scirrhous gastric cancer. The gels and the tissues were immunostained by a monoclonal antibody against human HSP47 and then examined by light and electron microscopy. RESULTS: The staining intensity of the fibroblasts was stronger than that of cancer cells in both the culture models and patient tissues. Moreover, the number of fibroblasts in scirrhous gastric cancer was significantly greater than that in non-scirrhous gastric cancer (p = 0.0004). In addition, the discharge of HSP47 was observed in the extracellular matrix as granular deposits of staining. CONCLUSION: These findings indicate that fibroblasts predominantly produce stromal collagen and may play an important role in the development of stromal change in scirrhous gastric cancer. Further studies are required to elucidate the significance of HSP47 in the development of new clinical approaches for treating scirrhous gastric cancer. PMID- 16475682 TI - Gallbladder small cell carcinoma Xenograft established by serial transplantation in nude mice. AB - The GB-04-JCK xenograft line of human gallbladder small cell carcinoma was established in nude mice by serial transplantation. The xenotransplantability has been maintained for more than 20 years. The carcinoma cells grew in a solid-sheet pattern and were found to have hyperchromatic nuclei, finely dispersed chromatin and inconspicuous nucleoli in the primary gallbladder tumor, as well as in the established xenograft GB-04-JCK The carcinoma cells also had Grimelius argyrophil granules, electron-dense neuroendocrine granules bounded by a single membrane. The xenograft line retained histological and immunohistochemical characteristics of the primary gallbladder tumor and is the first reported xenotransplantable tumor of human gallbladder small cell carcinoma. PMID- 16475683 TI - Effect of extracorporeal high hydrostatic pressure on cellular outgrowth from tumor-afflicted bone. AB - At present, in orthopedic surgery, the reconstruction of bone defects following resection of malignant tumors is effected by several methods. The irradiation and autoclaving of bone segments are the 2 methods of choice to extracorporeally devitalize the resected tumor-bearing bone segments. An alternative, gentle method of devitalizing bone-associated cells by exposing normal and tumor cells to extracorporeal high hydrostatic pressure (HHP) has been introduced. The aim of this study was to examine the ex vivo effect of HHP on the cell growth of normal and tumor-afflicted freshly-resected small human bone segments. For this, tumor afflicted human bone segments of 5 x 5 x 5 mm in size, obtained during surgery from 14 patients suffering from chondrosarcoma or osteosarcoma, in comparison to bone segments obtained from 36 patients with normal bone, disease were exposed to HHP levels of 0, 150 and 300 MPa for 10 min at 37 degrees C. Following HHP treatment, the specimens were placed into cell culture and observed for cell outgrowth up to 50 days. In control samples (0 MPa), rapid outgrowth of cells was observed. HHP-treatment of 150 MPa however, resulted in reduced outgrowth of cells from these bone specimens; at 300 MPa, no outgrowth of cells was detected. Light microscopy and standard histological examination showed morphological changes between control samples (0 MPa) and 150 MPa. Our results suggest that the treatment of tumor-afflicted bone and the associated cartilage by HHP leads to the devitalization of bone cells concomitant with complete impairment of cellular outgrowth, a precondition for re-implantation of the HHP-treated bone. PMID- 16475684 TI - Modification of pyrimidine derivatives from antiviral agents to antitumor agents. AB - 2,4-Diaminopyrimidine derivatives, that were originally developed as antiviral agents, were modified to antitumor agents by: (i) introducing an amino group at C 5 on the pyrimidine ring, (ii) changing the alkyl group and the ring size of the cycloalkyl group on the beta-position of the omega-hydroxyalkylamino group, (iii) replacing the phenylalkyl group on the cycloalkyl group with the 3,4,5 trimethoxyphenylalkyl group, (iv) the esterification of the primary alcohol with diethyl phosphate and (v) introducing the thiomethyl group at C-2 on the pyrimidine ring. Among the 21 compounds prepared, 6, which has cyclobutyl at the beta-position, exhibited potent activity towards P-388 leukemia. In addition, 14, with methoxyl groups on the phenyl ring and 17, with the thiomethyl group on the pyrimidine ring, showed specific inhibition for the EGFR protein kinase. Moreover, 15 and 16, which carry the diethyl phosphoryl group on the primary alcohol, exhibited inhibitory activity towards P-glycoprotein. PMID- 16475685 TI - Enforced expression of a truncated form of Bax-alpha (tBax) driven by human telomerase reverse transcriptase (hTERT) promoter sensitizes tumor cells to chemotherapeutic agents or tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). AB - BACKGROUND: We have recently demonstrated that a truncated form of the pro apoptotic molecule Bax-alpha (tBax) at the NH2-terminus is more potent in inducing cell death than wild-type (wt) Bax. In the present study, whether efficient cell death is induced by tBax expression from human telomerase reverse transcriptase (hTERT) promoter, which is highly active in tumor but not normal cells, was examined. MATERIALS AND METHODS: Cell death was assessed by luciferase reporter assay and the annexin staining method. RESULTS: Enforced expression of tBax resulted in cell death to a greater extent than wt Bax in two types of tumor cells: osteogenic sarcoma MG-63 and squamous cell carcinoma MIT7. The tBax sensitized these tumor cells to death induced by chemotherapeutic agents. Moreover, tBax enhanced cell death induced by the tumor necrosis factor-related apoptosis-inducing ligand to a high level, compared with wt Bax. Furthermore, tBax efficiently induced death of the MG-63 cells overexpressing Bcl-x(L), compared with wt Bax. CONCLUSION: tBax alone, or in combination with chemotherapeutic agents, would be a promising candidate for human gene therapy in the setting of carcinoma, especially for tumors containing high levels of Bcl x(L). PMID- 16475686 TI - Aberrant crypt foci. AB - Colon cancer evolves through epithelial cell deregulation and inappropriate proliferation. These histopathological characteristics are exemplified in the biochemical, immunohistochemical, genetic and epigenetic elements detected within colonic mucosa. Early detection is paramount for the prevention of colon cancer deaths. Aberrant crypt foci (ACF) are thought to be the earliest identifiable neoplastic lesions in the colon carcinogenetic model. The progression of ACF to polyp and, subsequently, to cancer parallels the accumulation of several biochemical alterations and mutations whereby a small fraction of ACF evolve to colon cancer. Recent data indicate that, not uncommonly, some ACF bypass the polyp stage in their carcinogenesis thus reinforcing the importance of their early detection and our understanding of their pathogenesis. Since ACF were first detected in carcinogen-treated mice, research efforts have focused on these microscopically visible lesions both in animal and human models. ACF show variable histological features, characterized by Kudo (20) and, therefore, can be grouped into differing categories by in vivo examination with high-magnification chromoscopic-colonoscopy (HMCC). As expected, ACF are more frequently detected in distal animal and human colons coinciding with the geographic distribution of colorectal cancer (CRC). Various proteomic (Prot) markers may be altered within ACF suggesting possible prospective pathological changes. These markers include Calreticulin, Transgelin, Serotransferrin, Triphosphate isomerase and Carbonic anhydrase II. Other markers of importance include carcinoembryonic antigen (CEA), B-catenin, placental cadherin (P-cadherin), epithelial cadherin (E-cadherin), inducible nitric oxide synthase (iNOS), cyclooxygenase (COX-2) and P16INK4a. Genetic mutations of K-ras, B-Raf APC and p53 have been demonstrated in ACF as well as the epigenetic alterations of CpG island methylation. Genomic instabilities (GI), illustrated by a higher GI Index (GII), microsatellite instability (MSI), loss of heterozygosity (LOH) and defects in mismatch repair (MMR) systems, are also expressed. These transformations may lead to the identification of the earliest pathological features initiating colon tumorigenesis. In this review, the advances in ACF research as precursors of CRCs are highlighted. PMID- 16475687 TI - Sphinganine causes early activation of JNK and p38 MAPK and inhibition of AKT activation in HT-29 human colon cancer cells. AB - The sphingoid base sphinganine induces apoptosis in HT-29 human colon cancer cells more potently than other bioactive sphingolipid metabolites sphingosine and C2-ceramide tested in our previous study. The objective of this study was to investigate the effect of sphinganine, at a concentration that induces apoptosis, on the mitogen activated protein kinases (MAPKs) including ERK1/ERK2, JNK2/JNK1, and p38 MAPK and AKT (protein kinase B), which regulate cell proliferation and apoptosis. HT-29 cells were cultured with sphinganine at 35 microM and the protein expression and phosphorylation status of ERK1/ERK2 (p44/p42), JNK2/JNK1 (p54/p46), p38 MAPK, and AKT were determined using Western blot analysis. Sphinganine clearly increased the active phosphorylated forms of JNK2/JNK1 and p38 MAPK after 15, 30, and 60 min treatment, with minimal effects on activation of ERK1/ERK2. Sphinganine weakly inhibited the phosphorylation of AKT at ser473 after 30 and 60 min. Sphinganine had little or no effect on the protein expression level of any of the kinases. The findings are consistent with a mechanism by which sphinganine induces apoptosis in HT-29 cells via early and strong activation of JNK and p38 MAPK and weak inhibition of AKT activation. PMID- 16475688 TI - Gefitinib decreases the synthesis of matrix metalloproteinase and the adhesion to extracellular matrix proteins of colon cancer cells. AB - BACKGROUND: Adhesion to extracellular matrix (ECM) proteins and degradation of basement membranes by matrix metalloproteinase (MMP) play important roles in cancer metastasis. In this study, the effects of gefitinib on the enzymatic activity of MMP and adhesion to ECM proteins in the HT29 colon cancer cell line were investigated. MATERIALS AND METHODS: Microtiter plates, coated with ECM proteins, were used to investigate the adhesion of cancer cells to ECM proteins. The expression of MMPs was examined by zymography and semiquantitative RT-PCR. RESULTS: Gefitinib inhibited MMP-9 and MMP-2 secretion and mRNA expression in HT29 cells. Gefitinib also reduced the ability to adhere to laminin and type IV collagen. These effects were observed at such low doses that gefitinib had neither an antiproliferative effect nor the ability to induce apoptosis. CONCLUSION: Gefitinib decreased the production of MMPs and the adhesion to ECM proteins, important steps associated with cancer metastasis. These results suggest that gefitinib may have antimetastatic activity in colon cancer. PMID- 16475689 TI - EGFR, c-erbB-2 and ki-67 in NSCLC and preneoplastic bronchial lesions. AB - BACKGROUND: The relationships between EGF-R and c-erbB-2 with other factors involved in tumour regulation are not well understood. The aim of this study was to correlate the expression of these markers with tumour proliferation. MATERIALS AND METHODS: The presence of EGF-R, c-erbB-2 and Ki-67 was evaluated by immunohistochemistry in non-small cell lung cancer (NSCLC) and preneoplastic lesions. RESULTS: Forty-two percent of the tumours were positive for EGF-R, 22% for c-erbB-2 and 97% for Ki-67. No statistically significant correlation was found between EGF-R and Ki-67, EGF-R and c-erbB-2 or between c-erbB-2 and Ki-67. With regards to Ki-67, a significant difference in survival was noted in favour of patients who did not express the marker. In preneoplastic lesions, most of the low-grade lesions showed neither EGF-R nor Ki-67 staining. In contrast, most of the high-grade lesions stained positively for these proteins. CONCLUSION: EGF-R and c-erbB-2 do not seem to be correlated with Ki-67 in NSCLC. PMID- 16475690 TI - Gemcitabine as a radiosensitizer in undifferentiated tumors. AB - BACKGROUND: Gemcitabine (dFdC) may cause radiosensitization by specific interference with homologous recombination-mediated DNA double-strand break repair. The radiosensitizing effect of dFdC might be less in normal healthy tissue and more restricted to undifferentiated tumor cells, making it a tumor selective radiosensitizer. Whether dFdC acts as a radiosensitizer in undifferentiated and well-differentiated rat tumors and on rat foot skin was tested. MATERIALS AND METHODS: Undifferentiated L44 lung tumors in BN rats, MLL prostate tumors in Copenhagen rats, and well-differentiated L42 lung tumors in WAG/Rij rats were used. The tumors were treated with a single X-ray dose, combined or not with dFdC (30 mg/kg) administered 24 h earlier. Tumor volume growth delay was the end-point used. In addition, rat foot skin was treated with a single dose of 22.5 Gy, with or without dFdC. The degree of skin damage was determined according to a scoring system. RESULTS: For tumor growth delay, the dose-enhancement ratios were 1.37 and 1.23-1.36 for the L44 and MLL tumors, respectively. No radiosensitization was observed for the well-differentiated L42 tumor and foot skin. CONCLUSION: Radiosensitization by dFdC was observed in the undifferentiated tumors, but not in the well-differentiated tumor and skin. Our data support further trials to evaluate the usefulness of dFdC as a radiosensitizer in undifferentiated tumors. PMID- 16475691 TI - Radiosensitization of CHO cells by two novel rhodium complexes under oxic and hypoxic conditions. AB - BACKGROUND: Tumour hypoxia severely limits the success of radiotherapy. Radiosensitization of hypoxic tumour cells by drugs is thus an important clinical issue. MATERIALS AND METHODS: Two novel ferrocene-containing beta-diketonato complexes of the transition metals rhodium and iridium were examined for their cytotoxic activity against Chinese hamster ovary (CHO) cells by MTT and clonogenic assays. The same complexes were also tested for their capacity to sensitize hypoxic CHO cells against 8 MeV photons. RESULTS: The IC50 for [Rh(fcta)(cod)] (I) and [Rh (fctca)(cod)] (II), where (fctfa) = ferrocenoylacetonato-4,4,4-trifluoro and (fctca) = ferrocenoyl-4,4,4-trichloro- and (cod) = 1,5-cyclooctadiene, were found to be 1.38 microM and 4.18 microM, respectively, closely resembling that of cisplatin which was found to be 1.21 microM. The rhodium (I) complex was identified as an effective anoxic radiosensitizer showing a dose-modifying factor (DMF) of 1.93 +/- 0.02, resembling cisplatin where the DMF was found to be 1.99 +/- 0.02. Small DMF's in the range of 1.10 were also found for cisplatin and the rhodium (I) complex under aerobic conditions, but these were not statistically significant. The DMF for the iridium complex was small and found to be 1.06 +/- 0.04. CONCLUSION: The cytotoxicity and radiosensitizing properties of the rhodium (I) complex are very similar to cisplatin and show considerable potential for clinical application. PMID- 16475692 TI - Limited suitability of EpCAM for molecular staging of tumor borders in head and neck cancer. AB - BACKGROUND: The epithelial cell adhesion molecule (EpCAM) is expressed in most normal epithelia, but is absent from squamous stratified epithelia. However, a de novo expression can be observed in squamous epithelia during carcinogenesis. MATERIALS AND METHODS: In order to evaluate EpCAM as a molecular marker to indicate borders of high risk for the development of local recurrences, its expression was examined in the marginal zone of malignancies. Specimens of squamous cell carcinoma of the head and neck (SCCHN), of the histologically tumor free defined resection margin and of healthy epithelia of 20 patients were examined by RT-PCR in order to identify the expression of EpCAM in these three different areas. Additionally, immunohistochemistry was perfonned on biopsies from 10 patients in order to confirm these findings and to investigate a potential correlation between EpCAM expression and the degree of dysplasia. RESULTS: By RT-PCR, high expression of EpCAM was found in the tumor. An inverse correlation was observed between EpCAM expression and the distance from the tumor, with no expression being detectable in healthy oral mucosa. In 70% of the cases, EpCAM was expressed in the marginal zone, which had been defined as tumor free by routine histopathological assessment. Additional immunohistology revealed no correlation between EpCAM expression and the grade of dysplasia. CONCLUSION: Our data provide evidence that EpCAM is restricted as a marker for redefining the real tumor margin by RT-PCR. To complement routine histology, immunohistochemical staining with EpCAM is limited due to its expression in hyperplastic tissue without dysplastic changes. Both observations limit the reliable use of EpCAM for the molecular definition of the critical tumor border and resection margins. PMID- 16475693 TI - Longitudinal evaluation of tumor metastasis by an FDG-microPet/microCT dual imaging modality in a lung carcinoma-bearing mouse model. AB - BACKGROUND: Histological methods are used to define the growth and response to various treatments of lung carcinoma in mice. The aim of the study was to evaluate a quantitative and 3D-tomographic microPET/microCT dual-image modality using 18F-fluorodeoxyglucose (FDG) to monitor the tumor progression in an experimental metastasis mouse model. MATERIALS AND METHODS: Six normal mice were subjected to FDG-microPET/microCT image scan to present the normal thorax morphology. Twenty-one 8-week-old male C57BL/6 mice were inoculated with 1 x 10(6) Lewis lung carcinoma cells (LLC1) through the lateral tail vein. FDG microPET/microCT scans were performed on days 0, 5, 9, 13 and 18 (n=6) to monitor the growth of the tumor. MicroPET and microCT images were further used to monitor the metastasis of the lung carcinoma to the liver. Fifteen mice were sacrificed for biodistribution on days 0, 5, 9, 13 and 18 after the inoculation of lung carcinoma cells. RESULTS: The FDG-microPET/microCT dual-image modality showed that the growth of the tumor could be monitored longitudinally. The standard uptake value (SUV) of FDG increased from 0.63 +/- 0.05 on day 0 to 1.03 +/- 0.15 on day 18, reflecting the growth of the tumor in mice. The tumors located in the lung and liver could be clearly visualized by the fusion of microPET and microCT images, and further confirmed by whole-body autoradiography or H&E stain. CONCLUSION: By FDG-microPET, the increase in SUV provided an alternative for assessing the growth of a tumor in vivo. Our results suggest that the growth progression of lung carcinoma can be identified using the FDG-microPET/microCT dual-image modality longitudinally in mice. PMID- 16475694 TI - Insulin-like growth factor 1 (IGF-1) and its receptor mRNA levels in breast cancer and adjacent non-neoplastic tissue. AB - Previous studies investigating the insulin-like growth factor 1 receptor (IGF-1R) expression in breast cancer tissue and adjacent non-neoplastic breast tissue (ANCT) have produced conflicting results. The IGF-1 and IGF-1R expression in pairs of breast cancer tissue and ANCT were investigated using RT-PCR and immunohistochemistry. The results of both methods were compared. MATERIALS AND METHODS: IGF-1 and IGF-1R mRNA from 31 specimen pairs were estimated using RT PCR. Immunohistochemistry for IGF-1R was carried out on 20 specimen pairs and the strength of staining was scored. RESULTS: The mean relative IGF-1 mRNA level was lower in the cancerous tissue (mean 0.450 +/- 0.206) than in the ANCT (mean 0.632 +/- 0.384) (paired t-test, p = 0.001). There was no measurable difference in relative IGF-1R mRNA levels in the cancerous tissue (mean 0.146 +/- 0.08) and the ANCT (mean 0.14608 +/- 0.108) (paired t-test, p = 0.807). Using immunohistochemistry, there was no statistical difference (paired t-test, p = 0.910) in IGF-1R staining scores between cancer (mean 1.93) and ANCT (mean 1.90). The comparison between the two methodologies showed no correlation (Pearson's Correlation Coefficient = -0.393). DISCUSSION: It can be concluded that IGF-1 expression is lower in cancerous tissue, thus supporting a paracrine relationship between cancerous tissue and ANCT, which may be useful in the prevention, diagnosis and treatment of breast cancer. There was no difference in the expression of the IGF-1 receptor in both types of tissue, as proven by RT-PCR and immunohistochemistty. Conflicting results in previous studies may be due to the different methods used to measure IGF-1R expression. PMID- 16475695 TI - Analysis of p53 protein by PAb240, Ki-67 expression and human papillomavirus DNA detection in different types of odontogenic keratocyst. AB - OBJECTIVE: To investigate the possible association between alterations in the p53 system and human papillomavirus (HPV) infection in the etiology of odontogenic keratocysts (OKCs) and to study proliferation and epithelial maturation patterns by topographic analysis of Ki-67 expression. MATERIALS AND METHODS: Eighty-three OKC samples (29 cases associated with nevoid basal cell carcinoma syndrome, 29 solitary non-recurrent cases 20 solitary recurrent cases, and 5 chondroid keratocysts) were studied by immunohistochemistry to detect p53 protein (PAb 244) and Ki-67 (MIB-1) expression, and by PCR to detect HPV DNA. RESULTS: Twelve cases (14.6%) expressed p53 protein; no case showed the presence of HPV DNA; 9 cases (11%) presented with mild epithelial dysplasia. The suprabasal expression of Ki 67 was significantly more frequent than its basal expression (p < 0.001). p53 protein expression was significantly associated with the presence of epithelial dysplasia (p = 0.023). Ki-67 expression was not associated with OKC type, the presence of dysplasia, or p53 expression. CONCLUSION: HPVs do not participate in the etiology of OKC, and it appears unlikely that a p53 gene mutation mechanism plays a major role in the genesis of OKC. OKCs show proliferation and genuine maturation behavior reminiscent of benign neoplasms with local destructive capacity. PMID- 16475696 TI - Apoptosis versus polemosis. Different mechanisms leading to non-necrotic cell death. AB - Apoptosis is a physiological auto-suicidal, genetically-induced cell deletion process of senescent effete normal cells. Apoptosis guarantees genetic fidelity, minimizes phenotypic variation and eliminates genotypic alteration. The auto destruction-induced by a cascade of caspases--results in the breakdown of normal cells and the formation of apoptotic bodies. Those bodies are rapidly phagocytized by macrophages and internalized by cells of the same type. On the other hand, polemosis (from Greek polemos: war) is a more dynamic mechanism of cell destruction between two different cell systems. Polemosis is focal, haphazardly distributed at the time of observation, probably erratic and influenced by the chemotactical attraction of committed lymphocytes by neoplastic cells. Polemosis is implemented by the interaction of the Fas-Fas ligand compulsory cellular system of self-defence. The end result of that struggle is manifested by the destruction of committed lymphocytes and the appearance of polemotic bodies (nuclear fragmentation with or without cytoplasmic remnants). The question arises as to whether polemotic bodies are also engulfed by macrophages, as is the case with apoptotic granules. In this work, the possible association between CD68-positive macrophages and intraepithelial polemotic bodies was investigated in sections from 50 colorectal adenomas. Polemotic bodies were found in groups of dysplastic cells in 84% (n=42) of the 50 adenomas, but none of them showed an accumulation of CD68 macrophages around the polemotic bodies. As polemotic bodies (autologous T-cells DNA) are not engulfed by macrophages it is hypothesized that the DNA of these bodies might be incorporated into the nuclei of dysplastic cells. This would satisfy the avidity of the DNA of rapidly proliferating dysplastic cells, a process that takes place, unremittingly, at any given time. PMID- 16475697 TI - Acridine orange used for photodynamic therapy accumulates in malignant musculoskeletal tumors depending on pH gradient. AB - BACKGROUND: Intra-operative photodynamic therapy has been applied with acridine orange (AO-PDT) to human musculoskeletal sarcomas for the past 4 years, resulting in a low local recurrence rate, within 10%, after intra-marginal tumor resection and excellent limb function. However, it is still unclear why acridine orange (AO) specifically accumulates in tumor cells, especially in malignant tumor cells. The purpose of this study was to clarify the mechanism of AO accumulation in malignant musculoskeletal tumors. MATERIALS AND METHODS: Sixty-two musculoskeletal tumors, including 35 malignant and 27 benign tumors, were studied. Using freshly resected tumor material, the extracellular pH (pHe) was measured and the fluorescence intensity of AO accumulated in the tumors was measured by an image analyzer after ex vivo exposure to 1.0 microg/ml AO, followed by blue excitation. In the in vitro study, bafilomycin A1 was exposed to LM 8 mouse osteosarcoma cells, in order to inhibit V-ATPase, subsequently causing a decrease in the pHgradient (deltapH) between the intracellular pH (pHi) and extracellular pH (pHe) or between the pHi and the pHe. AO accumulation and the cytocidal effect of AO were evaluated. RESULTS: The results of the in vivo study, using human materials freshly resected at surgery, revealed that the pHe of the malignant musculoskeletal tumors was significantly lower than that of the benign tumors and normal muscles or adipose tissues and also showed that the AO fluorescence intensity of the malignant musculoskeletal tumors was significantly stronger than that of the benign tumors and normal muscles or adipose tissues. The results also revealed that the AO fluorescence intensity negatively correlated with the pHe in tumors and normal tissues. The in vitro study showed that bafilomycin A1 inhibited the accumulation of AO in acidic organelles, such as lysosomes, and that the cytocidal effect of AO-PDT was also remarkably inhibited. DISCUSSION: Based on results of the in vivo and in vitro studies, it is suggested that malignant musculoskeletal tumors have a large deltapH between the pHi and the pHe or between the pHi and the vaculolar pH and also that a large ApH increases AO accumulation in tumors. We, therefore, believe that AO-PDT may be more effective in highly malignant musculoskeletal tumors than low grade ones, because of their acidity. CONCLUSION: AO accumulation in musculoskeletal tumors was dependent on the ApH between the pHi and the pHe, or between the pHi and the vacuolar pH. PMID- 16475699 TI - Proliferation measurement in breast cancer by two different methods. AB - BACKGROUND: Different studies show that proliferation measurement in breast cancer may have an independent prognostic value. In the present study, tumor proliferation in breast cancer was analyzed by two radically different methods according to the technique used (immunohistochemistry and flow cytometry), associated costs and necessary equipment. The aim was to evaluate which method discriminates better between tumors with high and low proliferation in relation to all other available clinical and biological parameters. MATERIALS AND METHODS: Two hundred and eighty breast cancers (231 ductal infiltrating, 30 lobular, 19 or less frequent varieties) were studied. The post-surgical staging was as follows: 164 pT1, 87 pT2, 7 pT3, the remaining 22 were multifocal, diffuse tumors. Axillary nodal invasion was found in 99 cases (35.4%). Proliferation was studied by means of flow cytometry (DNA index and S-phase) in fresh tumor tissue and immunohistochemistry (Ki67) in paraffin-embedded tissue. Furthermore, hormone receptor (estrogen receptor, ER; progesterone receptor, PR), c-erb-B2 and p53 expressions were studied using the same method. Finally, histological and nuclear grade, tumor size and axillary nodal invasion were also included as variables of the study. RESULTS: A DNA index >1 (aneuploidy) correlated significantly with histological grade 3 (p = 0.01), nuclear grade 3 (p < 0.0001), nodal invasion (p = 0.007), absence of ER (p = 0.006) and of PR (p = 0.002), c-erb-B2 expression (p = 0.008), p53 expression (p = 0.007) and tumor size (p = 0.01). An expression of Ki67 in 20% or more of tumor cell nuclei, on the other hand, correlated significantly with histological grade 3 (p < 0.0001), nuclear grade 3 (p < 0.0001), absence of ER (p < 0.0001) and of PR (p < 0.0001), c-erb-B2 expression (p < 0.0001), p53 expression (p < 0.0001) and tumor size (p = 0.0005), but not with nodal invasion. CONCLUSION: Although flow cytometry provides additional data (association with nodal invasion), the study of Ki67 expression emerges from this study as a simple, inexpensive and reliable method to study the proliferation rate of breast cancer. PMID- 16475698 TI - PTCH gene expression as a potential marker for esophageal squamous cell carcinoma. AB - BACKGROUND: The PTCH expression level in esophageal squamous cell carcinoma (SCC) was examined. MATERIALS AND METHODS: To test whether PTCH can serve as a candidate marker for esophageal SCC, a quantitative reverse transcription (RT) PCR for the PTCH gene was performed and the possible relationship between PTCH gene expression levels and clinicopathological findings in esophageal SCC was evaluated. RESULTS: A low PTCH expression score was observed in 18 out of 29 (62%) esophageal SCCs and it was significantly correlated with a poor prognosis in esophageal SCC patients (p = 0.0073). CONCLUSION: These results suggest that PTCH might serve as a new parameter for the prediction of prognosis in esophageal SCC. PMID- 16475700 TI - Ascorbyl stearate inhibits cell proliferation and tumor growth in human ovarian carcinoma cells by targeting the PI3K/AKT pathway. AB - Ascorbyl stearate is a lipophilic, vitamin C derivative with antitumorigenic properties. The molecular mechanism(s) underlying the anticarcinogenic effect of this compound have not been well documented. The effect of ascorbyl stearate was studied in a panel of human ovarian epithelial cancer cells. Treatment with ascorbyl stearate caused a dose-dependent inhibition of the cell proliferation. The antiproliferative effect was due to the arrest of cells in the S/G2-M-phase of the cell cycle. Treatment of OVCAR-3 cells with ascorbyl stearate also inhibited PI3K/AKT activity. The presence of a constitutively active AKT protected OVCAR-3 cells from the effects of ascorbyl stearate, suggesting that this nutraceutical targets the PI3K/AKT pathway. The administration of ascorbyl stearate by gavage induced involution of human ovarian carcinoma xenografts in nude mice. These studies indicate that the antiproliferative effect of ascorbyl stearate on ovarian epithelial cancer cells is associated with decreased PI3K/AKT activity, and point toward the PI3K/AKT signaling pathway as a target for this drug. PMID- 16475701 TI - Cytidine-phosphate-guanosine oligodeoxynucleotides and interferon-alpha expressing tumor cells effectively induce dendritic cell maturation in vitro. AB - BACKGROUND: Dendritic cells (DCs) play an important role in immune response and cytidine-phosphate-guanosine (CpG) oligodeoxynucleotides (ODN) as well as interferon (IFN)-alpha have been proven to induce DC maturation. In this study, the synergistic effects of CpG-ODN and IFN-alpha on DC maturation were evaluated. MATERIALS AND METHODS: Surface molecules on DCs and the stimulatory responses of DCs to allogeneic splenocytes were analyzed after cultivation with CpG-ODN and IFN-alpha-overexpressing murine colorectal cancer MC38 cells (MC38-IFNalpha). RESULTS: Co-incubation with CpG-ODN and MC38-IFNalpha, but not wild-type MC38 cells (MC38-WT), effectively up-regulated co-stimulatory molecules on the DCs. CpG, in combination with IFN-alpha, stimulated IL-1beta and TNF-alpha production by DCs effectively. When DCs preincubated with CpG-ODN and MC38-IFNalpha were co incubated with allogeneic splenocytes in vitro, the proliferation of these splenocytes was significantly enhanced compared with that of splenocytes incubated with CpG-ODN and MC38-WT cells (p = 0.041). CONCLUSION: Since CpG-ODN and IFN-alpha have synergistic effects on DC maturation, they may induce potent antitumor immune responses and combination therapy should be considered for clinical application. PMID- 16475702 TI - Diallyl disulfide inhibits WEHI-3 leukemia cells in vivo. AB - Enhanced garlic (Allium sativum) consumption is closely related to reduced cancer incidence, as shown in epidemiological studies. Diallyl disulfide (DADS), a component of garlic, inhibits the proliferation of human blood, colon, lung and skin cancer cells. Although DADS had been reported to induce apoptosis in human leukemia HL-60 cells, there are no reports regarding whether or not it affects leukemia cells in vivo. Therefore, the present study is focused on the in vivo effects of DADS on WEHI-3 leukemia cells. The effects of DADS on murine WEHI-3 cells were initially examined, and the results indicated that DADS induced cytotoxicity and that this effect was dose-dependent. The effects of DADS on WEHI 3 in BALBIc mice were also examined, and the results indicated that DADS decreased the percentage of MAC-3 marker, indicating that differentiation of the precursor of macrophage and T cells was inhibited. The weights of liver and spleen were also measured, and the results indicated that DADS decreased the weight of these organs. An important characteristic of WEHI-3 leukemia is the enlarged spleen in mice after i.p. injection of WEHI-3 cells. Based on pathological examination, the function of DADS was observed in the liver and spleen of mice previously injected with WEHI-3 cells. Apparently, DADS affects WEHI-3 cells both in vitro and in vivo. PMID- 16475703 TI - Apoptosis of human leukemia HL-60 cells and murine leukemia WEHI-3 cells induced by berberine through the activation of caspase-3. AB - The aim of this study was to clarify the mechanisms of apoptosis, cytotoxicity, DNA damage and fragmentation, as well as the production of reactive oxygen species (ROS) and Ca+2, induced by berberine in human promyelocytic leukemia HL 60 and murine myelomonocytic leukemia WEHI-3 cells. The levels of Bcl-2 and Bax, the changes of mitochondria membrane potential (MMP), cytochrome c release and activation of caspase-3 were also investigated in both cell lines. The flow cytometry and DAPI staining assays indicated that berberine induced cytotoxicity in both cell lines examined. Flow cytometry assay also showed that berberine induced ROS and Ca+2 production, decreased the levels of MMP and increased the activity of caspase-3 in both cell lines examined. Berberine-induced apoptosis was accompanied by increased levels of Ca+2 and a decrease in the mitochondrial membrane potential, leading to the release of cytochrome c and the cleavage of pro-caspase-3. Western blotting also showed that berberine increased the levels of Bax and cytochrome c and decreased the levels of Bcl-2 in both cell lines. Inhibition of caspase-3 activation (z-VAD-fmk: cell-permeable broad-spectrum caspase inhibitor) completely blocked berberine-induced apoptosis in both HL-60 and WEHI-3 cells. Therefore, berberine induced apoptosis in both examined cell lines through the activation of caspase-3. PMID- 16475704 TI - NSC 290205-based therapy in murine pancreatic adenocarcinoma PAN02 in combination with adriamycin (ADR). AB - BACKGROUND: NSC 290205 (A) is a hybrid synthetic antitumor ester, which combines a D-lactam derivative of androsterone and nitrogen mustard. In this study, the antitumor activity of A in combination with ADR (AHOP) was investigated in comparison with the standard CHOP regimen. MATERIALS AND METHODS: PAN02 adenocarcinoma was used in this study. C57Bl mice were used for chemotherapy evaluation. The activity was assessed from the inhibition of tumor growth and the oncostatic parameter T/C%. RESULTS: Treatment with A or cyclophosphamide produced almost equal borderline activity. Moreover, both the CHOP and AHOP regimens showed significant and comparable antitumor effects. AHOP caused the maximum effect, inhibiting tumor growth by 56.8%. CHOP was less effective, producing 47.7% tumor inhibition. CONCLUSION: It is very likely that the D-lactamic steroid (androstan) alkylator forA, containing the amide group -NH-CO-, combined with ADR which intercalates between DNA base-pairs, is the explanation for the higher activity of AHOP as compared to CHOP. PMID- 16475705 TI - Expression of Nm23 in gliomas and its effect on migration and invasion in vitro. AB - BACKGROUND: Despite the well-documented importance of Nm23 in the control of metastasis, there is currently a paucity of data regarding the role of this gene family in the control of glioma invasion. MATERIALS AND METHODS: Nm23-H1 expression in gliomas was assessed via immunohistochemistry, Western blot, RT-PCR and Northern blot analyses. The migration and invasion ability were also investigated in primary glioma culture cells, human glioma cell lines and nm23-H1 transfectant, using an in vitro brain slice invasion model and a simple scratch technique. RESULTS: Although no significant correlations were detected between nm23-H1 expression and pathological grade, the endogenous nm23-H1 expression in gliomas was found to be inversely correlated with their migratory abilities. Additionally, the nm23-H1 transfectant resulted in a reduction of approximately 45% of the migratory ability and suppressed the invasiveness of the parental cell line. CONCLUSION: Our overall findings suggest that nm23-H1 may play an important role in the suppression of glioma invasion and migration. PMID- 16475706 TI - Triptolide sensitizes resistant cholangiocarcinoma cells to TRAIL-induced apoptosis. AB - BACKGROUND: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL/Apo2L) promotes apoptosis by binding to transmembrane receptors. It is known to induce apoptosis in a wide variety of cancer cells, but TRAIL-resistant cancers have also been documented. In this study, the relative resistance of human cholangiocarcinoma (CCA) cell lines against TRAIL-induced apoptosis is reported and the possible potential synergistic effect with triptolide, a diterpene triepoxide extracted from the Chinese herb Tripterygium wilfordii, in killing TRAIL-resistant CCA cells is investigated. MATERIALS AND METHODS: Six human CCA cell lines were treated with various concentrations of TRAIL and the resistant cells were identified and subsequently tested for their sensitivity to a combination of TRAIL and triptolide. The susceptibility and resistance of the cells were based on analysis of cytotoxic and apoptotic induction and expression of anti-apoptotic factors (Mcl-1 and cFLIP). RESULTS: The treatment of TRAIL induced a dose-dependent decrease in cell viability in 4 out of the 6 cell lines. A combination of TRAIL and triptolide enhanced cytotoxicity and apoptosis in these 2 resistant cell lines. The combined treatment enhanced activation of caspase-8 and its downstream signaling processes compared with the treatment with either one alone. CONCLUSION: The results presented show that human CCA cells were heterogeneous with respect to susceptibility to TRAIL-induced apoptosis. The combination of TRAIL and triptolide could enhance susceptibility to TRAIL-induced apoptotic killing in these TRAIL-resistant CCA cells, thus offering an alternative approach for the treatment of TRAIL-resistant cholangiocarcinoma. PMID- 16475707 TI - 1H-NMR evidence for a different response to the same dose (2 Gy) of ionizing radiation of MG-63 human osteosarcoma cells and three-dimensional spheroids. AB - High resolution proton nuclear magnetic resonance (1H-NMR) spectroscopy was used to examine the response of the MG-63 osteosarcoma cell line grown in monolayer and as 3-dimensional tumor spheroids to the same low dose (2 Gy) of ionizing radiation. The MG-63 cells and spheroids were irradiated at 24 h of growth and the 1H-NMR spectra of whole control and irradiated monolayer cells and of whole control and irradiated multicellular spheroids collected after another 24 h were compared. The 1H-NMR spectra of the perchloric acid extracts as well as the 2 dimensional 1H-NMR spectra of both pairs of cell systems were also obtained. Possible radiation-induced cell damage was determined by lactate dehydrogenase (LDH) release and variations in cell growth, while cell death was evaluated by chromatin dye Hoechst staining and DNA fragmentation assays. The results demonstrated that no cell damage took place, but that significant variations in numerous metabolites occured in both the monolayer cells and the spheroids after irradiation. Most of the changes observed were very similar in nature. In fact, significant increases in lactate, alanine, creatine and phosphocreatine and choline-containing metabolites and a significant decrease in glutathione (GSH) were observed in both cells and spheroids. However, while significant increases in CH2 and CH3 mobile lipids, glutamine/glutamate, taurine and inositol were seen in the spheroids, no variations in CH2 or CH3 lipids, glutamine/glutamate or taurine were recorded in the MG-63 cells grown in monolayer after irradiation. In addition, a significant decrease rather than a significant increase in inositol was also noted in the monolayer cells. The data presented seem to suggest that, although neither monolayer cells nor spheroids show apparent signs of damage after exposure to the same dose of ionizing radiation, very different cell death responses as well as very diverse antioxidant/osmoregulatory reactions were triggered by this stressing agent. PMID- 16475708 TI - Bone-related growth factors and zoledronic acid regulate the PTHrP/PTH.1 receptor bioregulation systems in MG-63 human osteosarcoma cells. AB - Bisphosphonates are known to inhibit osteoclast-mediated bone resorption and osteoblast differentiation and are currently used in the treatment of Paget's disease, osteoporosis, metastatic and osteolytic bone disease and hypercalcaemia of malignancy. The parathyroid hormone-related peptide (PTHrP) and type 1 PTH/PTHrP receptor (PTH.1R) bioregulation systems mediate a wide range of local paracrine/autocrine and intracrine functions in various tissues and modify the actions of pharmaceutical agents on target tissues, both in vivo and in vitro. In addition, bone microenvironment-related growth factors, such as insulin-like growth factor 1 (IGF-1), transforming growth factor beta 1 (TGF beta 1), basic fibroblast growth factor (bFGF) and interleukin 6 (IL-6), can modify the actions of various pharmaceutical agents, including cytotoxic drugs in malignant cell lines. Whether IGF-1, TGF beta 1, bFGF, IL-6 and zoledronic acid affect the expressions of PTHrP and PTH.1R in MG-63 osteoblast-like osteosarcoma cells was investigated in this study. Relative quantitative-PCR (expression at mRNA level) and immunofluorescence analysis (localization of the expression at protein level) were employed to assess PTHrP and PTH.IR expressions. Our data showed that primarily IGF-1, TGF beta 1 and IL-6 (up to 25 ng/ml for 48 h) increased PTHrP mRNA expression and modified its perinuclear localization, while zoledronic acid (up to 100 microM for 48 h) inhibited cell proliferation and suppressed PTHrP expression in the MG-63 osteosarcoma cells. These growth factors were incapable of reversing the zoledronic acid decrease of the expression of PTHrP in the MG-63 cells, suggesting that zoledronic acid and the growth factors affect PTHrP expression via an independent intracellular signal transduction pathway in these cells. However, no appreciable modulation of the PTH.1R expression by IGF-1, TGF beta 1, bFGF, IL-6 or zoledronic acid was detected in MG-63 cells. Therefore, we conclude that PTHrP expression possibly mediates the action of bone microenvironment-related growth factors and of zoledronic acid in MG-63 cells. PMID- 16475709 TI - Bone morphogenetic protein-2 suppresses invasiveness of TSU-Pr1 cells with the inhibition of MMP-9 secretion. AB - Bone morphogenetic protein (BMP), which is a member of the transforming growth factor beta (TGF-beta) superfamily, has powerful osteoinductive effects and various biological activities in a variety of cells. The effect of BMP-2 on the human carcinoma cell line TSU-Pr1 was examined. BMP-2 was found to inhibit the migration and invasiveness, but not the proliferation, of TSU-Pr1 cells. Gel zymography for matrix metalloproteinases (MMPs) and urokinase-type plasminogen activator (uPA) detection revealed that BMP-2 down-regulated the activity of MMP 9, but not of MMP-2 or uPA. BMP-2 also reduced the secretion of MMP-9 into the culture media, but did not affect the secretion of MMP-2, Timp-1, Timp-2 or Timp 3. These results suggest that BMP-2 inhibits migration and decreases MMP-9 secretion to suppress the invasiveness of TSU-Pr1 cells. This is the first report of a role for BMP signaling in reducing the invasiveness of cancer cells. PMID- 16475710 TI - CYP2C9 polymorphisms in human tumors. AB - The oxazaphosphorines cyclophosphamide (CP) and ifosfamide (IF) are alkylating agents that require bioactivation via cytochrome (CYP) P450 isoenzymes including CYP2C9 enzymes. The present study investigated CYP2C9 in regard to its allelic variants in 23 tumor samples (10 breast tumors, 1 breast tumor cell line, 5 brain tumors, 7 glioma cell lines) with restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR). The mutant alleles of CYP2C9 were residue 144 (Arg (*1)/Cys (*2)), residue 358 (Tyr/Cys), residue 359 (Ile/Leu (*3)) and residue 417 (Gly/Asp). The frequencies of the CYP2C9*1, CYP2C9*2 and CYP2C9*3 alleles in the cancer samples examined were found to be 0.848, 0.152 and 0.043, respectively. No sample revealed a mutation at residue 358 or 417. Comparing breast with brain tumors, brain tumors seemed to reveal a higher incidence of heterozygotes (5/12 compared to 2/11) at residue 144 and, with regard to residue 359, a lower incidence of heterozygotes (0/12 compared to 2/11). In summary, our data indicate that in tumor material, as in healthy material, the same allelic variants of CYP2C9 occur. Compared to healthy tissue, tumor material seemed to reveal a higher incidence of the *2 allele, but a significant difference could not be established. Our results show that brain and breast tumor samples appeared to differ in their frequency of heterozygotes at residues 144 and 359. This might also have an impact on intratumoral oxazaphosphorine metabolism. PMID- 16475711 TI - In vivo antitumour effect of combretastatin A-4 phosphate added to fractionated irradiation. AB - BACKGROUND: The study aimed at evaluating the potential benefit from a combination of fractionated ionising radiation with the vascular-targeting compound combretastatin A-4 phosphate (CA-4-P). MATERIALS AND METHODS: Syngenic rat rhabdomyosarcoma (R1), growing subcutaneously, was treated at 2 different sizes: either small (2 +/- 0.5 cm3) or large (10.94 +/- 0.6 cm3). Localised fractionated irradiation of the tumours (5 x 3 Gy) in 5 days was followed 1 day later by an intraperitoneal CA-4-P treatment (25 mglkg). RESULTS: The combined treatment of only large tumours resulted in a small additional growth delay when compared with radiotherapy only. CONCLUSION: The present data indicate a size dependent increase in tumour growth delay from combining fractionated irradiation with CA-4-P. PMID- 16475712 TI - Recombinant adeno-associated and adenoviral vectors for the transduction of pancreatic and colon carcinoma. AB - BACKGROUND: Recombinant adeno-associated viral vectors serotype 2 (rAAV2) can transduce several tissues with high efficiency. MATERIALS AND METHODS: The transduction efficiency of rAAV2 in pancreatic and colon cancer was compared to that of recombinant adenovirus (rAd) in vitro and in vivo using green fluorescent protein (GFP). RESULTS: With the exception of SU.86.86, the percentage of GFP positive cells was below 10% at a multiplicity of infection (MOI) of 100 for rAAV2. At the same MOI, almost 100% of cells expressed GFP when rAd was used. However, the transduction efficiency for rAA V2 was comparable to that of rAd when coinfected with wt adenovirus, leading to a dramatic increase in the amount of double-stranded rAAV2 DNA. Similar results were obtained in vivo. While widespread GFP expression was readily detected in all xenografts injected with rAd, only one section of all tumors injected with rAAV2 contained GFP-positive cells. CONCLUSION: The results of this study indicate that rAAV2 might be useful for an ex vivo approach in cancer gene therapy, but it does not seem to be feasible for the in vivo treatment of malignant tumors. PMID- 16475713 TI - Sex chromosome abnormalities in bladder cancer: Y polysomies are linked to PT1 grade III transitional cell carcinoma. AB - BACKGROUND: Bladder cancer is a heterogeneous group of tumors from both the biological and clinical points of view. Conventional cytogenetics and molecular genetic techniques have shown non-random aberrations in bladder cancer, while certain chromosomal changes have been found to be highly correlated with tumor grade or stage. The aim of this study was to evaluate, by fluorescence in situ hybridization (FISH), the numerical aberrations of chromosomes X and Y in bladder cancer, comparing the incidence of nuclei with aneusomies in different grades or histological stages of the tumors. MATERIALS AND METHODS: The FISH technique, using DNA probes specific for chromosomes X and Y, was applied to 35 male bladder tumor specimens directly processed for cytogenetic study. RESULTS: Polysomies of chromosome X were observed in 25 out of the 35 cases examined (71.43%), while numerical aberrations of chromosome Y were observed in 22 out of the 35 cases (62.86%). Of those cases with numerical aberrations of chromosome Y, 13 had polysomy (37.14%), while in 9 cases, loss of Y was observed (25.71%). Statistical analysis showed that numerical aberrations on chromosomes X or Y were not linked to histological stage, while a probable correlation was observed between aneusomies X or Y and tumor grade. Comparing the results of PT1-grade III tumors with those of PT1-grade II, statistical analysis showed that aneusomies Y and, especially, polysomy Y were correlated with PT1-grade III tumors, p = 0.023 and p = 0.010, respectively. An uncertain correlation between polysomy X and PT1-grade III tumors was found, p = 0.070. CONCLUSION: Our results may suggest that the genetic instability associated with PT1-grade III tumors may account for the considerable potential for aggression of these tumors. However, to draw definite conclusions, a large number of cases must be studied. PMID- 16475714 TI - Primary lymphomas of bone. AB - Primary lymphomas of bone are uncommon malignancies. The vast majority of them are non-Hodgkin lymphoma (NHL), whereas primary Hodgkin lymphoma (HL) of bone is extremely rare. Patients with primary NHL of bone commonly present with local bone pain, soft tissue swelling, and a mass or a pathological fracture. There is a slight male preponderance, and most patients are over 45-50 years of age. Primary NHL of bone can arise in any part of the skeleton, but long bones (femurs, tibia) are the most common sites of presentation. Comprehensive immunohistochemical studies are required to establish an accurate histological diagnosis of primary NHL of bone. Most cases of primary NHL of bone are classified as diffuse large B-cell lymphomas (DLBCL) in the World Health Organisation (WHO) classification of hematological malignancies. On full staging evaluation, most patients have disease of stage IE or IIE according to the Ann Arbor system. Several studies indicate that patients with primary NHL of bone have a favorable outcome, especially when treated by combined modality therapy. A number of studies reported that clinical stage is the most important prognostic variable in predicting overall survival. Interestingly, the rare occurrence of primary lymphoma of bone is in contrast with the frequency of plasma cell tumors in bone. This could be due to the fact that, during normal B-cell differentiation, the bone marrow is the normal site of homing of plasma cells which are terminally-differentiated, immunoglobulin-secreting post-germinal center B-cells. In this respect, there is circumstancial evidence that primary NHL of bone may represent tumors of post-germinal center B-cells. The present review summarizes data on the histogenesis of primary NHL of bone in view of the recent histogenetic classification of DLBCL on the basis of the B-cell differentiation gene expression profiles (germinal center vs. post-germinal center B-cell differentiation). PMID- 16475715 TI - Induction of apoptosis in hematopoietic cells with an antibody against tomoregulin-1. AB - Tomoregulin-1, a type-I transmembrane protein with two follistatin modules, a unique epidermal growth factor (EGF) domain and a short, highly conserved cytoplasmic tail, was studied. A number of hematopoietic cell lines (L1210, CEM, Jurkat, U937, K562, JY, THP-1 and T2) express tomoregulin-1 endogenously. In these cells, apoptosis was induced by an antiserum (C29) and purified IgG against the follistatin modules, but not by antisera against the EGF-domain or the cytoplasmic tail. Furthermore, C29 induced apoptosis in tomoregulin-1-, but not in mock-transfected cells. Apoptosis was monitored through genomic DNA fragmentation, annexin-V staining and caspase-3 activation. Treatment of the cells with C29 in the presence of H89 (a SerlThr kinase inhibitor) or 8'-bromo cyclicAMP revealed that apoptosis was mediated by a cAMP-dependent Ser/Thr kinase. Moreover, C29 increased [cAMP]i over 5-fold. Together, these data suggest that the C29 antiserum against tomoregulin-1 induces apoptosis of hematopoietic cells. PMID- 16475716 TI - Evaluation of chemosensitivity of human bone marrow stromal cells--differences between common chemotherapeutic drugs. AB - BACKGROUND: Bone marrow stromal cells (BMSCs) are essential for normal hematopoiesis, but also support the growth and survival of malignant hematopoietic cells. There are currently no data available regarding the sensitivity of BMSCs to cytotoxic drugs widely used in the therapy of hematological diseases such as multiple myeloma or acute leukemia. MATERIALS AND METHODS: The chemosensitivity of the BMSC line HS-5 and of primary BMSCs from patients were evaluated in comparison to NCI-H929 myeloma and U937 leukemia cells. RESULTS: Both HS-5 cells and human BMSCs showed substantial cell death in response to chemotherapy. While alkylating agents (melphalan, treosulfan) and doxorubicin demonstrated marked BMSC toxicity, nucleotide analogs (gemcitabine, cytarabine) induced only limited BMSC apoptosis. CONCLUSION: Our data suggest that the BMSC toxicity of cytotoxic compounds should be considered in chemotherapeutic regimens. PMID- 16475718 TI - Importance of apoptotic mechanisms in inflammatory infiltrate of oral lichen planus lesions. AB - BACKGROUND: Oral lichen planus (OLP) is an autoimmune disease produced by T lymphocyte attack on basal epithelial cells. OLP can undergo malignant transformation that may be linked to an increased proliferative activity and decreased apoptosis rate of the epithelial cells, phenomena that may be influenced by the inflammatory infiltrate. The objectives of this study were to assess the quantitative importance of apoptosis in the inflammatory infiltrate of OLP and to discuss its influence on the persistence of this infiltrate and on the malignant transformation of this disease. MATERIALS AND METHODS: In 32 patients with OLP and 20 controls, apoptosis was studied by TUNEL assay and caspase-3 determination, while cell cycle arrest and senescence were studied by measurement of p21 expression. RESULTS: There was a low frequency of lymphocytic apoptosis according to both TUNEL (34.5% of cases negative, 65.5% with mild expression) and caspase-3 expression (42.9% of cases negative, 50% with mild expression) findings. P21 expression was also negative (9.7% of cases) or mild (80.6% of cases) in most cases. CONCLUSION: The absence or low rate of apoptosis observed in inflammatory cells in OLP appears to contribute to the persistence of the inflammatory infiltrate, potentiating the onset of molecular disorders in epithelial cells and favouring cancer development. PMID- 16475717 TI - TRAIL-induced cytotoxicity in a melphalan-resistant rhabdomyosarcoma cell line via activation of caspase-2. AB - The prognosis for patients with chemo-refractory rhabdomyosarcoma remains poor. The tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) is a hopeful candidate for new strategies in chemotherapy. The effects of TRAIL and melphalan (Mel) in the rhabdomyosarcoma cell line TE-671 were investigated by colorimetric caspase assays and flow cytometry. TRAIL induced the activation of caspases-2, -3 and -8, but not the activation of caspase-9, in the Mel-resistant TE-671 cells. Inhibition of caspase-2 with the caspase-2 inhibitor z-VDVAD-fmk significantly down-regulated the TRAIL-induced caspase-3 activation, as well as the TRAIL induced cytotoxicity. When TE-671 cells were treated with a combination of Mel and TRAIL, a significant synergism of drug-induced cytotoxicity was obtained. The inhibition of caspase-2 could completely abolish caspase-3 activation, suggesting that TRAIL sensitises TE-671 cells for Mel-induced cytotoxicity via a caspase-2- and -3-dependent mechanism. In conclusion, it was shown, for the first time, that TRAIL could sensitise Mel-resistant tumour cells to melphalan. PMID- 16475719 TI - 99mTc targeting of Sst2-expressing tumors by tetraamineoctreotide: first results in CA20948 cells and rat models. AB - BACKGROUND: Somatostatin subtype 2 receptor (sst2)-targeted tumor imaging with [(111)In-DTPA0]octreotide is an established method for scintigraphic detection of neuroendocrine tumors. [99mTc]Demotide ([99mTc-N4-bzdg0, Tyr3]octreotide), presented herein, is a potential sst2-targeting radiotracer based on 99mTc. MATERIALS AND METHODS: Demotide was synthesized in solution and labelled with 99mTc in alkaline medium. The in vitro properties of Demotide and [99mTc]Demotide were tested in sst2-expressing CA20948 cells and rat tissues, while biodistribution was studied in Lewis rats bearing CA20948 tumors. RESULTS: During labelling, [99mTc]Demotide was obtained in >98% yields and typical 1 mCi/nmol specific activities. Demotide showed in vitro sub-nM affinity for the sst2. While rapidly internalizing into CA20948 cells, [99mTc]Demotide localized effectively in CA20948 implants (2.2 +/- 1.1% ID/g at 1 h post injection) and in target organs in rats via a sst-mediated process and cleared rapidly via the kidneys. CONCLUSION: The above favorable characteristics validate [99mTc]Demotide as a promising 99mTc-radiotracer for the targeted-imaging of sst2-positive tumors. PMID- 16475720 TI - Reversal of multidrug resistance of cancer cells in vitro: modification of drug resistance by selected carotenoids. AB - The development of multidrug resistance (MDR) causes difficulties in the chemotherapy of of human cancer. Investigation of the possibility of reversal of MDR has been greatly aided by the use of cell lines with acquired resitance to anticancer agents in vitro or transfected with the mdrl gene. The aim of this study was to examine new perspectives of chemotherapy focused on natural, carotenoid compounds, in connection with the modification of MDR. The function of the MDR protein was examined via the R123 drug accumulation of both cell lines in the presence of carotenoids. The fluorescence of the cell population was measured by flow cytometry. The most effective resistance modifiers Monoepoxy-beta carotene, (SS, 8S)-capsochrome, (8'S) Luteoxanthin, (9Z)-Violaxanthin, (9Z) Zeaxanthin, (13Z)-Zeaxanthin were assayed for their antiproliferative effects in combination with the anti-cancer drug epirubicin. (13Z)-Zeaxanthin was able to enhance the antiproliferative effect on human mdrl gene transfected mouse lymphoma and anthracycline resistant human breast cancer cell line MCF7. (8'S) luteoxanthin, (5S, 8S)-capsochrome and (9Z)-zeaxanthin treatment revealed synergism with epirubicin on resistant mouse lymphoma. The enhanced antiproliferative activity of epirubicin combinated with (9Z)-Violaxanthin was more significant on MCF7 cells resistant to anthracycline. PMID- 16475721 TI - Differential expression of molecular markers in arsenic- and non-arsenic-related urothelial cancer. AB - BACKGROUND: Little is known about the mechanisms of arsenic-related urothelial cancer (AsUC). The aim of this study was to reveal the differential expression of molecular markers between AsUC and non-arsenic-related UC (non-AsUC). MATERIALS AND METHODS: Tissues of AsUC (n=33), non-AsUC (n=20) and normal bladder urothelia from patients with benign diseases (n=4) were examined for multiple selected molecular markers responsible for various cellular functions, includingglutathione, GST-pi, Bcl-2, p53 and c-Fos. RESULTS: The mean cellular glutathione content of normal mucosal samples (33.4 +/- 7.2 microM/mg protein) was significantly higher than either non-AsUC (22.8 +/- 1.8, p = 0.04) or AsUC (16.4 +/- 1.6, p = 0.002). The glutathione content of non-AsUC was higher than that of AsUC (p = 0.012). The expressions of Bcl-2 and c-Fos in AsUC were significantly higher than those in non-AsUC (p = 0.004 and p = 0.02, respectively). CONCLUSION: The carcinogenic pathway for AsUC is different, in part, from that of non-AsUC. Cellular glutathione contents may be down-regulated during urothelial carcinogenesis. Bcl-2 and c-Fos may play important roles in arsenic-mediated carcinogenesis of the urothelium. PMID- 16475723 TI - Influence of irinotecan and SN-38 on the irradiation response of WHO3 human oesophageal tumour cells under hypoxic conditions. AB - Irinotecan and its metabolite SN38 were evaluated for their cytotoxicity and influence on radiosensitivity in WHO3 human oesophageal cells under hypoxic conditions. The IC50's of Irinotecan and SN-38 were found to be 0.8 and 0.04 microM, repectively, with SN-38 emerging as the more potent drug. The toxicities were similar under anoxic conditions. Given in conjunction with irradiation under hypoxic conditions, the two drugs restored the radiosensitivity of WHO3 cells in a dose-dependent manner by factors of 1.5-2.1 as compared to a control oxygen enhancement ratio (OER) of 2.1 in this cell system. In the subtoxic concentration range of 10(-2) microM SN-38 still generated a marked sensitisation of hypoxic tumour cells by factors of 1.2-1.6. It is concluded that the topoisomerase inhibitor Irinotecan and in particular the metabolite SN-38 may be clinically useful for radiotherapy of notoriously hypoxic tumour pathologies. PMID- 16475722 TI - Polyphenols purified from the Brazilian aroeira plant (Schinus terebinthifolius, Raddi) induce apoptotic and autophagic cell death of DU145 cells. AB - Polyphenols extracted from many plants have shown antiproliferative and antitumor activities in a wide range of carcinogenesis models. The antiproliferative effects of polyphenols purified from the Brazilian aroeira plant (Schinus terebinthifolius, Raddi) were investigated on the androgen-insensitive DU145 human prostatic carcinoma cell line. A F3 fraction purified from leaf extract inhibited the DU145 cell proliferation more than 30-fold compared to the crude extract. By flow cytometric analysis, the polyphenol fraction was demonstrated to induce G0/G1 cell growth arrest and cell apoptosis. This apoptosis was evidenced by caspase 3 stimulation in F3-treated cells as compared to crude extract treated cells. The acid phosphatase activity of lysosomes was strongly activated in the lysosomal fraction of the F3-treated DU145 cells. This lysosomal activation, together with the appearance of autophagic vacuoles, suggests that "type 2 physiological cell death" was also involved in this antiproliferative effect. HPLC analysis of this F3 fraction showed 18 different subfractions. Among these subfractions, F3-3, F3-7 and F3-13 strongly inhibited DU145 cell proliferation in a dose-dependent manner. However, the nature of these polyphenols remains unknown since only one (Isoquercitrin) of the tested pure polyphenols co-migrated with F3 13. Since lysosomotropic drugs are considered as possible regulators of lysosome activity, aroeira polyphenols could target lysosomes of prostatic cancer cells to induce autophagic cell death. PMID- 16475724 TI - Cytological value of sputum in workers daily exposed to air pollution. AB - In this study, quantitative modifications of dust cells, siderocytes, Curschmann's spirals and asbestos bodies and qualitative modifications (cellular changes and inflammatory infiltrate) in the sputum of 164 traffic police officers and 218 railway workers, occupationally exposed to environmental pollution, and the sputum of 119 inhabitants of a rural area, were evaluated. The results were correlated with time of exposure and smoking habits. Seventy-three (45%) traffic police officers (TPO), 76 (35%) railway workers (RW) and 29 (24%) of the rural population (RP) were smokers. The sputum, collected over a 3-day period, was smeared on glass slides and stained according to the Papanicolaou, Perl and yellow eosin methods. The results of the qualitative cytological diagnosis revealed a statistically significant difference between the TPO, RW and the RP (p < 0.001). The results of the qualitative and quantitative cytological examinations were not significantly correlated to time of occupational exposure, which was considered to be a continuous variable. The qualitative cytological examination of sputa was not statistically significant for the smoking habits of the TPO and the RP, but was significant for the RW (p < 0.0067). In the TPO, the number of dust cells was higher in smokers, and the relative risk (RR) was 3.95. In the RW, the RR was 2.84. The results of our study revealed that for the RW, the qualitative-quantitative cytological alterations in sputum were due much more to smoking habits than to occupational exposure, while the presence of asbestos bodies correlated with work activity. The qualitative-quantitative cytological examinations of the TPO differed significantly from that of the other two populations. PMID- 16475725 TI - Reactions of OH-radicals with procarbazine. A pulse radiolysis and computer simulation study. AB - Pulse radiolysis was applied to study the reactivity of *OH radicals with procarbazine (PC), a cytostatic agent widely used in radiation- and chemotherapy. An overall rate constant of k(*OH+PC) = 3.7 x 10(9) l.mol(-1).s(-1) was determined. The thereby formed transients had a strong absorption at 350 nm, epsilon350 = 4.46 x 10(3) l.mol(-1).cm(-1), and a weak absorption band around 530 nm. Computer simulation studies to elucidate the most probable sites of *OH attack on the PC molecule showed that *OH radical addition to the aromatic ring had the highest probability. These transients decayed by a first order reaction, k = 1.75 x 10(3) s(-1), whereby species having a maximum absorption at 300 nm and broad shoulder at 340-380 nm were formed. Similar absorptions were observed after gamma radiolysis of PC. A reaction mechanism is suggested. For the reaction of H atoms with PC, a rate constant k(*H+PC) = 6.4 x 10(8) l.mol(-1).s(-1) was determined. PMID- 16475726 TI - Molecular staging of bladder cancer with RT-PCR assay for CK20 in peripheral blood, bone marrow and lymph nodes: comparison with standard histological staging. AB - BACKGROUND: The aim of this study was to analyze whether the CK20 reverse transcriptase polymerase chain reaction (RT-PCR) is suitable for detecting circulating tumor cells and residual tumor cells in lymph nodes, in patients with muscle invasive transitional cell carcinoma (TCC) of the bladder, and to compare these results with standard histological staging. PATIENTS AND METHODS: The nested RT-PCR assay was used to analyze the CK20 transcript in the peripheral blood, bone marrow, lymph nodes, the tumor and normal biopsies of bladder from 57 patients with invasive TCC of the bladder, who underwent radical cystectomy, and from 9 patients with noninvasive TCC. RESULTS: Lymph node pathological status was positive in 24 out of the 57 patients studied and all of them except I showed expression of CK20, with a correlation between histological technique and RT-PCR of 95.8%. A statistically significant correlation of lymph node CK20 RT-PCR with the standard risk factor of pathological stage (p = 0.04) was observed Blood and bone marrow CK20 RT-PCR showed no correlation with pathological stage. CONCLUSION: Lymph node CK 20 RT-PCR correlates with pathological stage in bladder cancer. The CK20 RT-PCR assay appears to be a highly sensitive and specific method for detecting circulating tumor cells and residual disease in lymph nodes in patients with invasive bladder cancer. Further evaluation of the significance of CK20 as a molecular marker for staging and follow-up in these patients is necessary. PMID- 16475728 TI - GSTT1 polymorphism and oral leukoplakia. AB - BACKGROUND: Considering that tobacco is the main etiological factor in oral cancer and that oral leukoplakia is the most prevalent potentially malignant lesion, together with the fact that GSTT1 polymorphism increases the risk of oral squamous cell carcinoma, the purpose of this study was to investigate the GSTT1 null polymorphism and the risk of oral leukoplakia in individuals with tobacco smoking habits in a Brazilian population. MATERIALS AND METHODS: The GSTT1 genotypes of 72 tobacco smoking patients with oral leukoplakia and 72 tobacco smoking healthy subjects were studied by PCR-based methods. RESULTS: The frequency of the GSTT1 null genotype in the group with oral leukoplakia (48.6%) was statistically different from the controls (27.8%). After stratification, the frequencies of the null genotype in female patients, lesions with absent/mild grade of dysplasia and from low-risk sites were statistically different from the control. CONCLUSION: The GSTT1 null genotype may increase the risk of developing oral leukoplakia. PMID- 16475727 TI - Circumvention of camptothecin-induced resistance during the adaptive cellular stress response. AB - Camptothecin-11 (CPT-11) induces the adaptive stress response in yeast, conferring resistance via not fully characterized mechanisms. This study aimed at exploring, pharmacologically, the mechanisms underlying the CPT-11-induced resistance in yeast. Post-logarithmic yeast cultures were submitted to heat shock following preconditioning with suramin and with CPT-11, either alone or in combination with suramin, cycloheximide, sodium molybdate, okadaic acid, or verapamil. The stress response was evaluated by determining cell viability after heat shock. Preconditioning with CPT-11 or suramin conferred thermotolerance to yeast cells. Co-administration of CPT-11 with suramin, cycloheximide or okadaic acid reversed the CPT-11-induced thermotolerant phenotype, while sodium molybdate and verapamil had no effect on CPT-11-induced resistance. The antagonistic effect of the thermotolerance-inducers and the possible contribution of topoisomerase II activity and post-translational modifications mediated by the phosphatases PP1/2A in CPT-11-induced resistance may have important implications on the acquisition of resistance to stress in eukaryotic cells. PMID- 16475730 TI - Evaluation of a series of new 99mTc-labeled bombesin-like peptides for early cancer detection. AB - Bombesin (BN) is a peptide exhibiting a high affinity for the gastrin-releasing peptide (GRP) receptor, which is overexpressed by a variety of tumors, including breast or prostate cancer. The aim of the present study was the investigation of the complexes formed between a series of BN-like peptides and the nuclides (185/187)Re and 99mTc. The (185/187)Re complexes were formed via the precursor Regluconate. The radiolabeling of the derivatives with 99mTc was performed using either 99mTc-gluconate or 99mTc-MDP as the intermediate complex. For the in vitro evaluation of the new peptides, the cancer cell line PC3 was used. The in vivo behavior of the 99mTc-labeled BN-like peptides was evaluated in normal mice. All the derivatives showed specific uptake in the pancreas, an organ rich in BN receptors and high affinity for the cancer cell line PC3. The above preliminary results indicated that the new BN derivatives are promising for human cancer studies. PMID- 16475729 TI - Comparative evaluation of linear and cyclic 99mTc-RGD peptides for targeting of integrins in tumor angiogenesis. AB - Cell adhesion molecules, such as integrins, play a vital role in angiogenesis, a key pathway for tumor growth, invasion and metastasis. The integrin alpha(v)beta(3), which recognizes the RGD sequence (Arg-Gly-Asp), may provide a target for in vivo tumor imaging. A linear and a cyclic RGD peptide derivative (RGDfK-His and cRGDfK-His, respectively), labelled via the precursor [99mTc(H2O)3(CO)3]+, were comparatively evaluated and their radiobiological properties were assessed in normal and tumor-bearing mice. Biodistribution studies showed non-specific uptake in all organs, rapid blood clearance and elimination via the hepatobiliary and urinary systems. Tumor uptake was higher for the cyclic radiolabelled derivative, as the both biodistribution and imaging studies suggested. The cRGDfK-His, labelled via the fac-[99mTc(CO)3]-core, may prove to be a useful tool for early tumor detection. PMID- 16475732 TI - Out with the old, in with the new. PMID- 16475731 TI - Heparin increases human gastric carcinoma cell growth. AB - BACKGROUND: Heparin has been widely used to prevent cancer-associated thromboembolism in cancer patients. Recent evidence reveals that heparin could modulate cell proliferation in the stomach. The effect of heparin on gastric cancer growth, however, is unknown. The effect of heparin on the proliferation of a human gastric adenocarcinoma cell line, BGC-823, was investigated. MATERIALS AND METHODS: Cell proliferation was assessed by [3H]-thymidine incorporation. The expressions of several growth-related genes were determined by RT-PCR and Western blot. RESULTS: Heparin significantly increased cell proliferation in BGC-823 cancer cells by 15.5% at the dose of 0.2 microg/ml. Heparin also up-regulated c Myc protein expression by 14.4%. In contrast, mRNA and protein levels of epidermal growth factor receptor (EGFR) were, respectively, down-regulated by 12.7% and 8.2% with no effect on cyclooxygenase-2 mRNA or protein expression. CONCLUSION: Our results suggest that heparin can promote the proliferation and up regulation of c-Myc protein expression in gastric cancer cells. PMID- 16475733 TI - Interoperability: what it means, why it matters. PMID- 16475734 TI - The future of patient identification. PMID- 16475735 TI - Beyond data sharing. For physicians, optimism and caution over using networked patient data. PMID- 16475736 TI - A quick scan of bar coding. When planning document imaging systems, don't forget the bar codes. PMID- 16475737 TI - Experts: clinical terminology mapping will be essential tool. PMID- 16475738 TI - Reports: Health IT can address healthcare fraud. PMID- 16475739 TI - Consumers moving to the center of NHIN planning. PMID- 16475740 TI - Keeping online personal records private: security and privacy considerations for Web-based PHR systems. PMID- 16475741 TI - Medical record completion. PMID- 16475742 TI - Long-term care's IT agenda. Industry summit produces 11 action items for health IT adoption. PMID- 16475743 TI - Surveying the RHIO landscape. A description of current RHIO models, with a focus on patient identification. AB - Across the country, new RHIOs are being formed every day. The 21 RHIOs studied by the work group illustrate the variety of purposes, funding, and record linking methods RHIOs may adopt. As this trend continues to evolve and improve, RHIOs may prove to be a valuable stepping stone on the road to a national system in which a patient's medical data will be available anywhere, anytime. Accurate patient identification and linking are the foundation of health technology that is implemented in a RHIO or any similar network that shares patient information. Without accurate patient identification, patient safety and quality of care are compromised. When high percentages of duplication or overlaying of records occurs in electronic health record databases, physician trust in the system is lost. As HIM professionals, we must be involved in addressing the security and confidentiality of RHIO databases and in defining the record linking method appropriate to the RHIO. As professionals skilled in patient identification methods and possessing significant organizational skills and personnel management experience, HIM professionals should become involved in this process at the earliest opportunity in the RHIO formation. HIM professionals can participate in long-term planning, business plan development, and organizational structure definition. Future articles will address how HIM professionals can become involved, what particular attributes and skills they can bring to the table, and job descriptions appropriate to HIM professionals in the healthcare information sharing industry. The work group urges all HIM professionals to become involved personally in this exciting new field. PMID- 16475744 TI - You can't throw IT in the dumpster anymore. Tips for finding a qualified electronic waste disposal service. PMID- 16475745 TI - 2006 CPT coding update. PMID- 16475746 TI - The codes for chronic kidney disease. Help in distinguishing between renal failure and renal insufficiency. PMID- 16475747 TI - Advocating for patients. Critical care nurse makes the move to e-HIM future. PMID- 16475748 TI - Methods for image segmentation should be standardized and calibrated. For the proposition. PMID- 16475749 TI - Methods for image segmentation should be standardized and calibrated. Against the proposition. PMID- 16475750 TI - Benchmarking analytical calculations of proton doses in heterogeneous matter. AB - A proton dose computational algorithm, performing an analytical superposition of infinitely narrow proton beamlets (ASPB) is introduced. The algorithm uses the standard pencil beam technique of laterally distributing the central axis broad beam doses according to the Moliere scattering theory extended to slablike varying density media. The purpose of this study was to determine the accuracy of our computational tool by comparing it with experimental and Monte Carlo (MC) simulation data as benchmarks. In the tests, parallel wide beams of protons were scattered in water phantoms containing embedded air and bone materials with simple geometrical forms and spatial dimensions of a few centimeters. For homogeneous water and bone phantoms, the proton doses we calculated with the ASPB algorithm were found very comparable to experimental and MC data. For layered bone slab inhomogeneity in water, the comparison between our analytical calculation and the MC simulation showed reasonable agreement, even when the inhomogeneity was placed at the Bragg peak depth. There also was reasonable agreement for the parallelepiped bone block inhomogeneity placed at various depths, except for cases in which the bone was located in the region of the Bragg peak, when discrepancies were as large as more than 10%. When the inhomogeneity was in the form of abutting air-bone slabs, discrepancies of as much as 8% occurred in the lateral dose profiles on the air cavity side of the phantom. Additionally, the analytical depth-dose calculations disagreed with the MC calculations within 3% of the Bragg peak dose, at the entry and midway depths in the phantom. The distal depth-dose 20%-80% fall-off widths and ranges calculated with our algorithm and the MC simulation were generally within 0.1 cm of agreement. The analytical lateral-dose profile calculations showed smaller (by less than 0.1 cm) 20%-80% penumbra widths and shorter fall-off tails than did those calculated by the MC simulations. Overall, this work validates the usefulness of our ASPB algorithm as a reasonably fast and accurate tool for quality assurance in planning wide beam proton therapy treatment of clinical sites either composed of homogeneous materials or containing laterally extended inhomogeneities that are comparable in density and located away from the Bragg peak depths. PMID- 16475751 TI - Optimized interstitial PDT prostate treatment planning with the Cimmino feasibility algorithm. AB - The primary aim of this study was to determine whether optimized photodynamic therapy (PDT) treatment planning (seeking optimized positions, lengths, and strengths of the light sources to satisfy a given dose prescription) can improve dose coverage to the prostate and the sparing of critical organs relative to what can be achieved by the standard PDT plan. The Cimmino algorithm and search procedures based on that algorithm were tested for this purpose. A phase I motexafin lutetium (MLu)-mediated photodynamic therapy protocol is ongoing at the University of Pennsylvania. PDT for the prostate is performed with cylindrical diffusing fibers of various lengths inserted perpendicular to a base plate to obtain longitudinal coverage by a matrix of parallel catheters. The standard plan for the protocol uses sources of equal strength with equal spaced (1-cm) loading. Uniform optical properties were assumed. Our algorithms produce plans that cover the prostate and spare the urethra and rectum with less discrepancy from the dose prescription than the standard plan. The Cimmino feasibility algorithm is fast enough that changes to the treatment plan may be made in the operating room before and during PDT to optimize light delivery. PMID- 16475752 TI - The UF series of tomographic computational phantoms of pediatric patients. AB - Two classes of anthropomorphic computational phantoms exist for use in Monte Carlo radiation transport simulations: tomographic voxel phantoms based upon three-dimensional (3D) medical images, and stylized mathematical phantoms based upon 3D surface equations for internal organ definition. Tomographic phantoms have shown distinct advantages over the stylized phantoms regarding their similarity to real human anatomy. However, while a number of adult tomographic phantoms have been developed since the early 1990s, very few pediatric tomographic phantoms are presently available to support dosimetry in pediatric diagnostic and therapy examinations. As part of a larger effort to construct a series of tomographic phantoms of pediatric patients, five phantoms of different ages (9-month male, 4-year female, 8-year female, 11-year male, and 14-year male) have been constructed from computed tomography (CT) image data of live patients using an IDL-based image segmentation tool. Lungs, bones, and adipose tissue were automatically segmented through use of window leveling of the original CT numbers. Additional organs were segmented either semiautomatically or manually with the aid of both anatomical knowledge and available image-processing techniques. Layers of skin were created by adding voxels along the exterior contour of the bodies. The phantoms were created from fused images taken from head and chest-abdomen-pelvis CT exams of the same individuals (9-month and 4 year phantoms) or of two different individuals of the same sex and similar age (8 year, 11-year, and 14-year phantoms). For each model, the resolution and slice positions of the image sets were adjusted based upon their anatomical coverage and then fused to a single head-torso image set. The resolutions of the phantoms for the 9-month, 4-year, 8-year, 11-year, and 14-year are 0.43 x 0.43 x 3.0 mm, 0.45 x 0.45 x 5.0 mm, 0.58 x 0.58 x 6.0 mm, 0.47 X 0.47 x 6.00 mm, and 0.625 x 0.625 x 6.0 mm, respectively. While organ masses can be matched to reference values in both stylized and tomographic phantoms, side-by-side comparisons of organ doses in both phantom classes indicate that organ shape and positioning are equally important parameters to consider in accurate determinations of organ absorbed dose from external photon irradiation. Preliminary studies of external photon irradiation of the 11-year phantom indicate significant departures of organ dose coefficients from that predicted by the existing stylized phantom series. Notable differences between pediatric stylized and tomographic phantoms include anterior-posterior (AP) and right lateral (RLAT) irradiation of the stomach wall, left lateral (LLAT) and right lateral (RLAT) irradiation of the thyroid, and AP and posterior-anterior (PA) irradiation of the urinary bladder. PMID- 16475753 TI - Target volume dose considerations in proton beam treatment planning for lung tumors. AB - We performed a treatment planning study in order to gather basic insight in the effect of setup errors and breathing motion on the cumulative proton dose to a lung tumor. We used a simplified geometry that simulates a 50 mm diameter gross tumor volume (GTV) located centrally inside lung tissue. The GTV was expanded with a uniform 5 mm margin into a clinical target volume (CTV) and into a variety of planning target volume (PTV's). Proton beam apertures were designed to conform the prescribed dose laterally to the PTV while the range compensator was designed to provide distal coverage of the CTV. Different smearing distances were applied to the range compensators, and the cumulative dose in the CTV was evaluated for different combinations of breathing motion and systematic setup errors. Evaluation parameters were the dose to 99% of the CTV (D99) and the equivalent uniform dose (EUD), with a surviving fraction at 2 Gy of SF2 = 0.5. For a single proton field designed to a 15 mm expansion of the CTV and without smearing applied to the range compensator, D99 of the CTV reduced from 96% for no tumor displacement to 41% and 13% for systematic setup errors of 5 and 10 mm, respectively. For a representative clinical combination, of 5 mm systematic error and 10 mm breathing amplitude, the EUD of the CTV was about 40 Gy (prescribed dose 70 Gy) regardless the CTV to PTV margin, and without smearing. Smearing the range compensator increases the dose to the CTV substantially with a lateral margin and smearing distance of 7.5 mm providing ample tumor coverage. In this latter case, D99 of the target volume increased to 87% for a single field treatment plan. Smearing does, however, lead to an increase in dose to normal tissues distal to the clinical target volume. Next to countering geometric mismatches due to patient setup, smearing can also be used to counter the detrimental effects of breathing motion on the dose to the clinical target volume. We show that the lateral margin and smearing distance can be substantially smaller than the maximum tumor displacement due to setup errors and patient breathing, as measured by the D99 and the EUD. PMID- 16475754 TI - Correction of pixel sensitivity variation and off-axis response for amorphous silicon EPID dosimetry. AB - The aim of this work is to determine the pixel sensitivity variation and off-axis dose response of an amorphous silicon electronic portal imaging device (EPID), and develop a correction method to improve EPID dosimetry. The uncorrected or raw pixel response of the aS500 amorphous silicon EPID shows differences in response (sensitivity) of individual pixels as well as a large off-axis differential response with respect to an ion chamber in water. Both can be corrected by division of raw images by the flood-field (FF) image. However, this leads to two problems for dosimetry: (1) the beam profile is present in both the raw image and FF image, and hence is "washed out" of the corrected image, and (2) any mismatch of EPID position between dosimetry and FF calibration means that the beam profile and off-axis response in the raw image and FF are misaligned. This causes artifacts in FF division and dosimetric errors. A method was developed to measure the off-axis response and pixel sensitivity variation separately to allow correction of images at any EPID position while retaining beam profile information. The pixel sensitivity variation is applied to the imager plane and is independent of imager position. The off-axis response depends on the imager plane position relative to the beam central axis. The pixel sensitivities were derived from multiple images of the same symmetric field acquired with the detector displaced laterally between each image. The off-axis response was measured by acquiring off-axis raw images (FF correction removed) and dividing out the off-axis beam fluence and previously determined pixel sensitivity differences. The dosimetric errors due to lateral and vertical detector displacement with the conventional FF calibration method were measured and compared to the new method. Corrected EPID profiles were then compared to beam profiles measured with ion chamber in water for open fields. The EPID was found to have a large off-axis differential response with respect to an ion chamber in water, particularly for 6 MV. This increased to 13% at 15 cm off-axis for 6 MV, and 3.5% for 18 MV at the isocenter plane. The dosimetric errors introduced by detector displacement with conventional FF calibration were found to be approximately 1% per centimeter of lateral detector displacement and 0.1% per centimeter of vertical displacement. These were reduced to less than 1% for any position with the new correction method. Corrected EPID images agreed with ion chamber measurements to within 2% (excluding penumbra and low-dose areas outside the field) for various field sizes. The new correction method gives consistent dosimetry for any EPID position and retains beam profile information in the image. PMID- 16475756 TI - Monte Carlo simulation estimates of neutron doses to critical organs of a patient undergoing 18 MV x-ray LINAC-based radiotherapy. AB - Absorbed photoneutron dose to patients undergoing 18 MV x-ray therapy was studied using Monte Carlo simulations based on the MCNPX code. Two separate transport simulations were conducted, one for the photoneutron contribution and another for neutron capture gamma rays. The phantom model used was of a female patient receiving a four-field pelvic box treatment. Photoneutron doses were determinate to be higher for organs and tissues located inside the treatment field, especially those closest to the patient's skin. The maximum organ equivalent dose per x-ray treatment dose achieved within each treatment port was 719 microSv/Gy to the rectum (180 degrees field), 190 microSv/Gy to the intestine wall (0 degrees field), 51 microSv/Gy to the colon wall (90 degrees field), and 45 microSv/Gy to the skin (270 degrees field). The maximum neutron equivalent dose per x-ray treatment dose received by organs outside the treatment field was 65 microSv/Gy to the skin in the antero-posterior field. A mean value of 5 +/- 2 microSv/Gy was obtained for organs distant from the treatment field. Distant organ neutron equivalent doses are all of the same order of magnitude and constitute a good estimate of deep organ neutron equivalent doses. Using the risk assessment method of the ICRP-60 report, the greatest likelihood of fatal secondary cancer for a 70 Gy dose is estimated to be 0.02% for the pelvic postero anterior field, the rectum being the organ representing the maximum contribution of 0.011%. PMID- 16475755 TI - Expandable and rigid endorectal coils for prostate MRI: impact on prostate distortion and rigid image registration. AB - Endorectal coils (ERCs) are used for acquiring high spatial resolution magnetic resonance (MR) images of the human prostate. The goal of this study is to determine the impact of an expandable versus a rigid ERC on changes in the location and deformation of the prostate gland and subsequently on registering prostate images acquired with and without an ERC. Sagittal and axial T2 weighted MR images were acquired from 25 patients receiving a combined MR imaging/MR spectroscopic imaging staging exam for prostate cancer. Within the same exam, images were acquired using an external pelvic phased array coil both alone and in combination with either an expandable ERC (MedRad, Pittsburgh, PA) or a rigid ERC (USA Instruments, Aurora, OH). Rotations, translations and deformations caused by the ERC were measured and compared. The ability to register images acquired with and without the ERC using a manual rigid-body registration was assessed using a similarity index (SI). Both ERCs caused the prostate to tilt anteriorly with an average tilt of 18.5 degrees (17.4 +/- 9.9 and 19.5 +/- 11.3 degrees, mean +/- standard deviation, for expandable and rigid ERC, respectively). However, the expandable coil caused a significantly larger distortion of the prostate as compared to the rigid coil; compressing the prostate in the anterior/posterior direction by 4.1 +/- 3.0 mm vs 1.2 +/- 2.2 mm (14.5% vs 4.8%) (p < 0.0001), and widening the prostate in the right/left direction by 3.8 +/- 3.7 mm vs 1.5 +/- 3.1 mm (8.3% vs 3.4%) (p = 0.004). Additionally, the ability to manually align prostate images acquired with and without ERC was significantly (p < 0.0001) better for the rigid coil (SI = 0.941 +/- 0.008 vs 0.899 +/- 0.033, for the rigid and expandable coils, respectively). In conclusion, the manual rigid-body alignment of prostate MR images acquired with and without the ERC can be improved through the use of a rigid ERC. PMID- 16475757 TI - The physics of computed radiography: measurements of pulse height spectra of photostimulable phosphor screens using prompt luminescence. AB - Computed radiography (CR) is a digital technology that employs reusable photostimulable phosphor (PSP) imaging plates (IP) to acquire radiographic images. In CR, the x-ray attenuation pattern of the imaged object is temporarily stored as a latent charge image within the PSP. The latent image is optically readout as photostimulated luminescence (PSL) when the phosphor is subsequently stimulated using a scanning laser. The multiple stages necessary to create a CR image make it difficult to investigate either experimentally or theoretically. In order to examine the performance of the CR system at a fundamental level separate measurements of the processes involved are desirable. Here pulse height spectroscopy is used to study the prompt violet light emission or prompt luminescence (PL) from commercial PSP screens. Since the mechanism by which light escapes from the phosphor is identical for PL and PSL, observations and conclusions based on the pulse height spectra (PHS) of PL are relevant to the understanding of the behavior of the PSL light emission that outputs the radiographic image in CR. The PL PHS of screens of different thickness and optical properties were measured and compared with the PHS of conventional phosphors. A new method for calibration of the PHS in terms of the absolute number of optical photons per x-ray is introduced and compared to previously established methods. PMID- 16475758 TI - Important changes in medical x-ray imaging facility shielding design methodology. A brief summary of recommendations in NCRP Report No. 147. AB - The recently published Report No. 147 of The National Council on Radiation Protection and Measurements entitled "Structural shielding design for medical x ray imaging facilities" provides an update of shielding recommendations for x rays used for medical imaging. The goal of this report is to ensure that the shielding in these facilities limits radiation exposures to employees and members of the public to acceptable levels. Board certified medical and health physicists, as defined in this report, are the "qualified experts" who are competent to design radiation shielding for these facilities. As such, physicists must be aware of the new technical information and the changes from previous reports that Report No. 147 supersedes. In this article we summarize the new data, models and recommendations for the design of radiation barriers in medical imaging facilities that are presented in Report No. 147. PMID- 16475759 TI - Reduction of false positives by internal features for polyp detection in CT-based virtual colonoscopy. AB - In this paper, we present a computer-aided detection (CAD) method to extract and use internal features to reduce false positive (FP) rate generated by surface based measures on the inner colon wall in computed tomographic (CT) colonography. Firstly, a new shape description global curvature, which can provide an overall shape description of the colon wall, is introduced to improve the detection of suspicious patches on the colon wall whose geometrical features are similar to that of the colonic polyps. By a ray-driven edge finder, the volume of each detected patch is extracted as a fitted ellipsoid model. Within the ellipsoid model, CT image density distribution is analyzed. Three types of (geometrical, morphological, and textural) internal features are extracted and applied to eliminate the FPs from the detected patches. The presented CAD method was tested by a total of 153 patient datasets in which 45 patients were found with 61 polyps of sizes 4-30 mm by optical colonoscopy. For a 100% detection sensitivity (on polyps), the presented CAD method had an average FPs of 2.68 per patient dataset and eliminated 93.1% of FPs generated by the surface-based measures. The presented CAD method was also evaluated by different polyp sizes. For polyp sizes of 10-30 mm, the method achieved mean number of FPs per dataset of 2.0 with 100% sensitivity. For polyp sizes of 4-10 mm, the method achieved 3.44 FP per dataset with 100% sensitivity. PMID- 16475760 TI - The effects of the imaging system on the validity limits of the ray-optical approach to phase contrast imaging. AB - A theoretical analysis of the x-ray phase contrast imaging and its validation via synchrotron radiation imaging is here presented. Two different mathematical models have been followed: the simpler ray-optical approach and the more rigorous Fresnel-Kirchoff diffraction theory. Subsequently, the conditions upon which the x-ray optical approximation can be used to describe the image formation mechanism have been analyzed, taking into account also the effects due to the finite source size and detector resolution. It is possible to demonstrate that the ray-optics results can also be obtained by opportunely developing the diffraction formalism only with some restrictions on the spatial frequencies present in the final image, without any limitation on the maximum phase shift. The conditions allowing the use of the simplified ray-optical approach to describe the phase contrast images have been here defined and their validation has been proved by means of computer simulations and phantom experiments. PMID- 16475761 TI - Calculation and validation of the use of effective attenuation coefficient for attenuation correction in In-111 SPECT. AB - Nuclear medicine tracers using 111In as a radiolabel are increasing in their use, especially in the domain of oncologic imaging. In these applications, it often is critical to have the capability of quantifying radionuclide uptake and being able to relate it to the biological properties of the tumor. However, images from single photon emission computed tomography (SPECT) can be degraded by photon attenuation, photon scattering, and collimator blurring; without compensation for these effects, image quality can be degraded, and accurate and precise quantification is impossible. Although attenuation correction for SPECT is becoming more common, most implementations can only model single energy radionuclides such as 99mTc and 123I. Thus, attenuation correction for 111In is challenging because it emits two photons (171 and 245 keV) at nearly equal rates (90.2% and 94% emission probabilities). In this paper, we present a method of calculating a single "effective" attenuation coefficient for the dual-energy emissions of 111In, and that can be used to correct for photon attenuation in radionuclide images acquired with this radionuclide. Using this methodology, we can derive an effective linear attenuation coefficient Micro(eff) and an effective photon energy E(eff) based on the emission probabilities and linear attenuation coefficients of the 111In photons. This approach allows us to treat the emissions from 111In as a single photon with an effective energy of 210 keV. We obtained emission projection data from a tank filled with a uniform solution of 111In. The projection data were reconstructed using an iterative maximum likelihood algorithm with no attenuation correction, and with attenuation correction assuming photon energies of 171, 245, and 210 keV (the derived E(eff)). The reconstructed tomographic images demonstrate that the use of no attenuation correction, or correction assuming photon energies of 171 or 245 keV introduces inaccuracies into the reconstructed radioactivity distribution when compared against the effective energy method. In summary, this work provides both a theoretical framework and experimental methodology of attenuation correction for the dual-energy emissions from 111In. Although these results are specific to 111In, the foundation could easily be extended to other multiple-energy isotopes. PMID- 16475762 TI - Determination of penumbral widths from ion chamber measurements. AB - We have investigated methods of reconstructing beam profiles in the penumbral region using a set of axially symmetric chambers, differing only in the detector radius. In principal, the transfer functions, or kernels, of such chambers should be functions of radius only. Three chambers of radii 0.297, 0.556, and 0.714 cm have been used. The transfer functions of the chambers can be determined by deconvolving the profiles measured with each detector with the PPMC profile. The results indicate that the transfer functions can be parametrized accurately as a Gaussian cutoff at 1.75(r), with (r) the average radius of the chamber. Deconvolution of the measured profiles with the transfer functions yields a profile that agrees with the PPMC profile to +/- 0.5 mm over the 20-80% penumbra. PMID- 16475763 TI - Gated CT imaging using a free-breathing respiration signal from flow-volume spirometry. AB - Respiration-induced tumor motion is known to cause artifacts on free-breathing spiral CT images used in treatment planning. This leads to inaccurate delineation of target volumes on planning CT images. Flow-volume spirometry has been used previously for breath-holds during CT scans and radiation treatments using the active breathing control (ABC) system. We have developed a prototype by extending the flow-volume spirometer device to obtain gated CT scans using a PQ 5000 single slice CT scanner. To test our prototype, we designed motion phantoms to compare image quality obtained with and without gated CT scan acquisition. Spiral and axial (nongated and gated) CT scans were obtained of phantoms with motion periods of 3-5 s and amplitudes of 0.5-2 cm. Errors observed in the volume estimate of these structures were as much as 30% with moving phantoms during CT simulation. Application of motion-gated CT with active breathing control reduced these errors to within 5%. Motion-gated CT was then implemented in patients and the results are presented for two clinical cases: lung and abdomen. In each case, gated scans were acquired at end-inhalation, end-exhalation in addition to a conventional free-breathing (nongated) scan. The gated CT scans revealed reduced artifacts compared with the conventional free-breathing scan. Differences of up to 20% in the volume of the structures were observed between gated and free-breathing scans. A comparison of the overlap of structures between the gated and free breathing scans revealed misalignment of the structures. These results demonstrate the ability of flow-volume spirometry to reduce errors in target volumes via gating during CT imaging. PMID- 16475764 TI - Radiation dose reduction in four-dimensional computed tomography. AB - Four-dimensional (4D) CT is useful in many clinical situations, where detailed abdominal and thoracic imaging is needed over the course of the respiratory cycle. However, it usually delivers a larger radiation dose than the standard three-dimensional (3D) CT, since multiple scans at each couch position are required in order to provide the temporal information. Our purpose in this work is to develop a method to perform 4D CT scans at relatively low current, hence reducing the radiation exposure of the patients. To deal with the increased statistical noise caused by the low current, we proposed a novel 4D penalized weighted least square (4D-PWLS) smoothing method, which can incorporate both spatial and phase information. The 4D images at different phases were registered to the same phase via a deformable model, thereby, a regularization term combining temporal and spatial neighbors can be designed for the 4D-PWLS objective function. The proposed method was tested with phantom experiments and a patient study, and superior noise suppression and resolution preservation were observed. A quantitative evaluation of the benefit of the proposed method to 4D radiotherapy and 4D PET/CT imaging are under investigation. PMID- 16475765 TI - Fractal characterization of brain lesions in CT images. AB - Fractal Dimension (FD) is a parameter used widely for classification, analysis, and pattern recognition of images. In this work we explore the quantification of CT (computed tomography) lesions of the brain by using fractal theory. Five brain lesions, which are portions of CT images of diseased brains, are used for the study. These lesions exhibit self-similarity over a chosen range of scales, and are broadly characterized by their fractal dimensions. PMID- 16475766 TI - Optimization of radiotherapy dose-time fractionation with consideration of tumor specific biology. AB - The "four Rs" of radiobiology play an important role in the design of radiation therapy treatment protocol. The purpose of this work is to explore their influence on external beam radiotherapy for fast and slowly proliferating tumors and develop an optimization framework for tumor-biology specific dose-time fractionation scheme. The linear quadratic model is used to describe radiation response of tumor, in which the time dependence of sublethal damage repair and the redistribution and reoxygenation effects are included. The optimum radiotherapeutic strategy is defined as the treatment scheme that maximizes tumor biologically effective dose (BED) while keeping normal tissue BED constant. The influence of different model parameters on total dose, overall treatment time, fraction size, and intervals is also studied. The results showed that, for fast proliferating tumors, the optimum overall time is similar to the assumed kickoff time T(k) and almost independent of interval patterns. Significant increase in tumor control can be achieved using accelerated schemes for the tumors with doubling time smaller than 3 days, but little is gained for those with doubling time greater than 5 days. The incomplete repair of normal tissues between two consecutive fractions in standard fractionation has almost no influence on the fractional doses, even for the hyperfractionation with an interval time of 8 h. However, when the resensitization effect is included, the fractional doses at the beginning and end of each irradiated week become obviously higher than others in the optimum scheme and the hyperfractionation scheme has little advantage over the standard or hypofractionation one. For slowly proliferating tumors, provided that the alpha/beta ratio of the tumor is comparable to that of the normal tissues, a hypofractionation is more favorable. The overall treatment time should be larger than a minimum, which is predominantly determined by the resensitization time. The proposed technique provides a useful tool to systematically optimize radiotherapy for fast and slow proliferating tumors and sheds important insight into the complex problem of dose-time fractionation. PMID- 16475767 TI - A novel method for automatic detection of patient out-of-plane rotation by comparing a single portal image to a reference image. AB - A novel method for detecting out-of-plane patient rotation by comparing a single portal image to its reference image is presented. Out-of-plane rotation results in an apparent distortion of the anatomy in a portal image. This distortion can be mathematically predicted with the magnification varying at each point in the image. While scaling of points at equal depth is invariant under in-plane rotation or translation, and changes equally in both dimensions for an axial shift of the patient, a change of scaling in only one dimension can be ascribed to an out-of-plane rotation. For the two conditions that are used in this study, it is shown that out-of-plane rotation yields a different scaling of the image in two perpendicular directions and therefore it is feasible to calculate the scale factors as a function of out-of-plane rotation. Conversely the recovery of scale factors in two different directions at the same time would enable the magnitude of the out-of-plane rotation to be recovered. The properties of the Fourier transform of the image are used to align the portal image with the reference image (a simulator image or first approved portal image) prior to the recovery of the scale factors. Correlating the Fourier transform of the portal image on a log scale with that of the reference image enables the scale factors to be automatically extracted from a single portal image. In the two approaches investigated, out-of-plane rotations of up to 41 degrees and 20 degrees (respectively) have been recovered with a maximum error of 2.4 degrees. This technique could be used to automatically detect patient roll or tilt prior to or during a treatment session. PMID- 16475768 TI - MCPI: a sub-minute Monte Carlo dose calculation engine for prostate implants. AB - An accelerated Monte Carlo code [Monte Carlo dose calculation for prostate implant (MCPI)] is developed for dose calculation in prostate brachytherapy. MCPI physically simulates a set of radioactive seeds with arbitrary positions and orientations, merged in a three-dimensional (3D) heterogeneous phantom representing the prostate and surrounding tissue. MCPI uses a phase space data source-model to account for seed self-absorption and seed anisotropy. A "hybrid geometry" model (full 3D seed geometry merged in 3D mesh of voxels) is used for rigorous treatment of the interseed attenuation and tissue heterogeneity effects. MCPI is benchmarked against the MCNP5 code for idealized and real implants, for 103Pd and 125I seeds. MCPI calculates the dose distribution (2-mm voxel mesh) of a 103Pd implant (83 seeds) with 2% average statistical uncertainty in 59 s using a single Pentium 4 PC (2.4 GHz). MCPI is more than 10(3) and 10(4) times faster than MCNP5 for prostate dose calculations using 2- and 1-mm voxels, respectively. To illustrate its usefulness, MCPI is used to quantify the dosimetric effects of interseed attenuation, tissue composition, and tissue calcifications. Ignoring the interseed attenuation effect or slightly varying the prostate tissue composition may lead to 6% decreases of D100, the dose delivered to 100% of the prostate. The presence of calcifications, covering 1%-5% of the prostate volume, decreases D80, D90, and D100 by up to 32%, 37%, and 58%, respectively. In conclusion, sub-minute dose calculations, taking into account all dosimetric effects, are now possible for more accurate dose planning and dose assessment in prostate brachytherapy. PMID- 16475769 TI - Liver segmentation for CT images using GVF snake. AB - Accurate liver segmentation on computed tomography (CT) images is a challenging task especially at sites where surrounding tissues (e.g., stomach, kidney) have densities similar to that of the liver and lesions reside at the liver edges. We have developed a method for semiautomatic delineation of the liver contours on contrast-enhanced CT images. The method utilizes a snake algorithm with a gradient vector flow (GVF) field as its external force. To improve the performance of the GVF snake in the segmentation of the liver contour, an edge map was obtained with a Canny edge detector, followed by modifications using a liver template and a concavity removal algorithm. With the modified edge map, for which unwanted edges inside the liver were eliminated, the GVF field was computed and an initial liver contour was formed. The snake algorithm was then applied to obtain the actual liver contour. This algorithm was extended to segment the liver volume in a slice-by-slice fashion, where the result of the preceding slice constrained the segmentation of the adjacent slice. 551 two-dimensional liver images from 20 volumetric images with colorectal metastases spreading throughout the livers were delineated using this method, and also manually by a radiologist for evaluation. The difference ratio, which is defined as the percentage ratio of mismatching volume between the computer and the radiologist's results, ranged from 2.9% to 7.6% with a median value of 5.3%. PMID- 16475770 TI - High-speed photography during ultrasound illustrates potential therapeutic applications of microbubbles. AB - Ultrasound contrast agents consist of microscopically small encapsulated bubbles that oscillate upon insonification. To enhance diagnostic ultrasound imaging techniques and to explore therapeutic applications, these medical microbubbles have been studied with the aid of high-speed photography. We filmed medical microbubbles at higher frame rates than the ultrasonic frequency transmitted. Microbubbles with thin lipid shells have been observed to act as microsyringes during one single ultrasonic cycle. This jetting phenomenon presumably causes sonoporation. Furthermore, we observed that the gas content can be forced out of albumin-encapsulated microbubbles. These free bubbles have been observed to jet, too. It is concluded that microbubbles might act as a vehicle to carry a drug in gas phase to a region of interest, where it has to be released by diagnostic ultrasound. This opens up a whole new area of potential applications of diagnostic ultrasound related to targeted imaging and therapeutic delivery of drugs such as nitric oxide. PMID- 16475771 TI - An improved analytical model for CT dose simulation with a new look at the theory of CT dose. AB - Gagne [Med. Phys. 16, 29-37 (1989)] has previously described a model for predicting the sensitivity and dose profiles in the slice-width (z) direction for CT scanners. The model, developed prior to the advent of multidetector CT scanners, is still widely used; however, it does not account for the effect of anode tilt on the penumbra or include the heel effect, both of which are increasingly important for the wider beams (up to 40 mm) of contemporary, multidetector scanners. Additionally, it applied only on (or near) the axis of rotation, and did not incorporate the photon energy spectrum. The improved model described herein transcends all of the aforementioned limitations of the Gagne model, including extension to the peripheral phantom axes. Comparison of simulated and measured dose data provides experimental validation of the model, including verification of the superior match to the penumbra provided by the tilted-anode model, as well as the observable effects on the cumulative dose distribution. The initial motivation for the model was to simulate the quasiperiodic dose distribution on the peripheral, phantom axes resulting from a helical scan series in order to facilitate the implementation of an improved method of CT dose measurement utilizing a short ion chamber, as proposed by Dixon [Med. Phys. 30, 1272-1280 (2003)]. A more detailed set of guidelines for implementing such measurements is also presented in this paper. In addition, some fundamental principles governing CT dose which have not previously been clearly enunciated follow from the model, and a fundamental (energy-based) quantity dubbed "CTDI-aperture" is introduced. PMID- 16475772 TI - An international dosimetry exchange for boron neutron capture therapy. Part I: Absorbed dose measurements. AB - An international collaboration was organized to undertake a dosimetry exchange to enable the future combination of clinical data from different centers conducting neutron capture therapy trials. As a first step (Part I) the dosimetry group from the Americas, represented by MIT, visited the clinical centers at Studsvik (Sweden), VTT Espoo (Finland), and the Nuclear Research Institute (NRI) at Rez (Czech Republic). A combined VTT/NRI group reciprocated with a visit to MIT. Each participant performed a series of dosimetry measurements under equivalent irradiation conditions using methods appropriate to their clinical protocols. This entailed in-air measurements and dose versus depth measurements in a large water phantom. Thermal neutron flux as well as fast neutron and photon absorbed dose rates were measured. Satisfactory agreement in determining absorbed dose within the experimental uncertainties was obtained between the different groups although the measurement uncertainties are large, ranging between 3% and 30% depending upon the dose component and the depth of measurement. To improve the precision in the specification of absorbed dose amongst the participants, the individually measured dose components were normalized to the results from a single method. Assuming a boron concentration of 15 microg g(-1) that is typical of concentrations realized clinically with the boron delivery compound boronophenylalanine-fructose, systematic discrepancies in the specification of the total biologically weighted dose of up to 10% were apparent between the different groups. The results from these measurements will be used in future to normalize treatment plan calculations between the different clinical dosimetry protocols as Part II of this study. PMID- 16475773 TI - 2D-3D registration of coronary angiograms for cardiac procedure planning and guidance. AB - We present a completely automated 2D-3D registration technique that accurately maps a patient-specific heart model, created from preoperative images, to the patient's orientation in the operating room. This mapping is based on the registration of preoperatively acquired 3D vascular data with intraoperatively acquired angiograms. Registration using both single and dual-plane angiograms is explored using simulated but realistic datasets that were created from clinical images. Heart deformations and cardiac phase mismatches are taken into account in our validation using a digital 4D human heart model. In an ideal situation where the pre- and intraoperative images were acquired at identical time points within the cardiac cycle, the single-plane and the dual-plane registrations resulted in 3D root-mean-square (rms) errors of 1.60 +/- 0.21 and 0.53 +/- 0.08 mm, respectively. When a 10% timing offset was added between the pre- and the intraoperative acquisitions, the single-plane registration approach resulted in inaccurate registrations in the out-of-plane axis, whereas the dual-plane registration exhibited a 98% success rate with a 3D rms error of 1.33 +/- 0.28 mm. When all potential sources of error were included, namely, the anatomical background, timing offset, and typical errors in the vascular tree reconstruction, the dual-plane registration performed at 94% with an accuracy of 2.19 +/- 0.77 mm. PMID- 16475774 TI - Dosimetric characteristics of a new unshielded silicon diode and its application in clinical photon and electron beams. AB - Shielded p-silicon diodes, frequently applied in general photon-beam dosimetry, show certain imperfections when applied in the small photon fields occurring in stereotactic or intensity modulated radiotherapy (IMRT), in electron beams and in the buildup region of photon beam dose distributions. Using as a study object the shielded p-silicon diode PTW 60008, well known for its reliable performance in general photon dosimetry, we have identified these imperfections as effects of electron scattering at the metallic parts of the shielding. In order to overcome these difficulties a new, unshielded diode PTW 60012 has been designed and manufactured by PTW Freiburg. By comparison with reference detectors, such as thimble and plane-parallel ionization chambers and a diamond detector, we could show the absence of these imperfections. An excellent performance of the new unshielded diode for the special dosimetric tasks in small photon fields, electron beams and build-up regions of photon beams has been observed. The new diode also has an improved angular response. However, due to its over-response to low-energy scattered photons, its recommended range of use does not include output factor measurements in large photon fields, although this effect can be compensated by a thin auxiliary lead shield. PMID- 16475775 TI - Contrast cancellation technique applied to digital x-ray imaging using silicon strip detectors. AB - Dual-energy mammographic imaging experimental tests have been performed using a compact dichromatic imaging system based on a conventional x-ray tube, a mosaic crystal, and a 384-strip silicon detector equipped with full-custom electronics with single photon counting capability. For simulating mammal tissue, a three component phantom, made of Plexiglass, polyethylene, and water, has been used. Images have been collected with three different pairs of x-ray energies: 16-32 keV, 18-36 keV, and 20-40 keV. A Monte Carlo simulation of the experiment has also been carried out using the MCNP-4C transport code. The Alvarez-Macovski algorithm has been applied both to experimental and simulated data to remove the contrast between two of the phantom materials so as to enhance the visibility of the third one. PMID- 16475776 TI - Technique factors and their relationship to radiation dose in pendant geometry breast CT. AB - The use of breast computed tomography (CT) as an alternative to mammography in some patients is being studied at several institutions. However, the radiation dosimetry issues associated with breast CT are markedly different than in the case of mammography. In this study, the spectral properties of an operational breast CT scanner were characterized both by physical measurement and computer modeling of the kVp-dependent spectra, from 40 to 110 kVp (Be window W anode with 0.30 mm added Cu filtration). Previously reported conversion factors, normalized glandular dose for CT-DgN(ct), derived from Monte Carlo methods, were used in concert with the output spectra of the breast scanner to compute the mean glandular dose to the breast based upon different combinations of x-ray technique factors (kVp and mAs). The mean glandular dose (MGD) was measured as a function of the compressed breast thickness (2-8 cm) and three different breast compositions (0%, 50%, and 100% glandular fractions) in four clinical mammography systems in our institution. The average MGD from these four systems was used to compute the technique factors for breast CT systems that would match the two-view mammographic dose levels. For a 14 cm diameter breast (equivalent to a 5 cm thick compressed breast in mammography), air kerma levels at the breast CT scanner's isocenter (468 mm from the source) of 4.4, 6.4, and 9.0 mGy were found to deliver equivalent mammography doses for 0%, 50%, and 100% glandular breasts (respectively) at 80 kVp. At 80 kVp (where air kerma was 11.3 mGy/100 mAs at the isocenter), 57 mAs (integrated over the entire scan) was required to match the mammography dose for a 14 cm 50% glandular breast. At 50 kVp, 360 mAs is required to match mammographic dose levels. Tables are provided for both air kerma at the isocenter and mAs for 0%, 50%, and 100% glandular breasts. Other issues that impact breast CT technique factors are also discussed. PMID- 16475777 TI - Preliminary study on the use of nonrigid registration for thoraco-abdominal radiosurgery. AB - The inclusion of organ deformation and movement in radiosurgery treatment planning is of increasing importance as research and clinical applications begin to take into consideration the effects of physiological processes, like breathing, on the shape and position of lesions. In this scenario, the challenge is to localize the target in toto (not only by means of marker sampling) and to calculate the dose distribution as the sum of all the contributions from the positions assumed by the target during the respiratory cycle. The aim of this work is to investigate the use of nonrigid registration for target tracking and dynamic treatment planning, i.e., treatment planning based not on one single CT scan but on multiple CT scans representative of the respiration. Twenty patients were CT scanned at end-inhale and end-exhale. An expert radiation oncologist identified the PTV in both examinations. The two CT data sets per patient were nonrigidly registered using a free-form deformation algorithm based on B-splines. The optimized objective function consisted of a weighted sum of a similarity criterion (Mutual Information) and a regularization factor which constrains the transformation to be locally rigid. Once the transformation was obtained and the registration validated, its parameters were applied to the target only. Finally, the deformed target was compared to the PTV delineated by the radiation oncologist in the other study. The results of this procedure show an agreement between the center of mass as well as volume of the target identified automatically by deformable registration and manually by the radiation oncologist. Moreover, obtained displacements were in agreement with body structure constraints and considerations usually accepted in radiation therapy practice. No significant influence of initial target volume on displacements was found. In conclusion, the proposed method seems to offer the possibility of using nonrigid registrations in radiosurgery treatment planning, even if more cases need to be investigated in order to give a statistical consistency to parameter setup and proposed considerations. PMID- 16475778 TI - Feasibility of four-dimensional conformal planning for robotic radiosurgery. AB - Organ motion can have a severe impact on the dose delivered by radiation therapy, and different procedures have been developed to address its effects. Conventional techniques include breath hold methods and gating. A different approach is the compensation for target motion by moving the treatment beams synchronously. Practical results have been reported for robot based radiosurgery, where a linear accelerator mounted on a robotic arm delivers the dose. However, not all organs move in the same way, which results in a relative motion of the beams with respect to the body and the tissues in the proximity of the tumor. This relative motion can severely effect the dose delivered to critical structures. We propose a method to incorporate motion in the treatment planning for robotic radiosurgery to avoid potential overdosing of organs surrounding the target. The method takes into account the motion of all considered volumes, which is discretized for dose calculations. Similarly, the beam motion is taken into account and the aggregated dose coefficient over all discrete steps is used for planning. We simulated the treatment of a moving target with three different planning methods. First, we computed beam weights based on a 3D planning situation and simulated treatment with organ motion and the beams moving synchronously to the target. Second, beam weights were computed by the 4D planning method incorporating the organ and beam motion and treatment was simulated for beams moving synchronously to the target. Third, the beam weights were determined by the 4D planning method with the beams fixed during planning and simulation. For comparison we also give results for the 3D treatment plan if there was no organ motion and when the plan is delivered by fixed beams in the presence of organ motion. The results indicate that the new 4D method is preferable and can further improve the overall conformality of motion compensated robotic radiosurgery. PMID- 16475779 TI - Patient specific treatment verifications for helical tomotherapy treatment plans. AB - We performed two-dimensional treatment verifications for ten patients planned and treated with helical tomotherapy. The treatment verification consisted of a film measurement as well as point dose measurements made with an ion chamber. The agreement between the calculated and the measured film dose distributions was evaluated with the gamma index calculated for three sets of criteria (2 mm and 2%, 4 mm and 3%, and 3 mm and 5%) as recommended in the literature. Good agreement was found between measured and calculated distributions without any need of normalization of the dose data but with dose map registration using reference marks. In this case, 69.8 +/- 17.2%, 92.6 +/- 9.0%, and 93.4 +/- 8.5% passed the 2 mm and 2%, 4 mm and 3%, and 3 mm and 5% criteria, respectively. Agreement was excellent when both normalization and manual registration of the dose maps was employed. In this case 91.2 +/- 5.6%, 99.0 +/- 1.4%, and 99.5 +/- 0.8% passed the 2 mm and 2%, 4 mm and 3%, and 3 mm and 5% criteria, respectively. The mean percent discrepancy for the point dose measurements was -0.5 +/- 1.1%, 2.4 +/- 3.7%, -1.1 +/- 7.3% for the high dose, low dose, and critical structure point, respectively. Three criteria for a satisfactory treatment verification in the high dose regions of a plan were established. For the un-normalized reference mark registered data 80% of pixels must pass the 3 mm and 5% criteria. For the normalized and manually registered data, 80% must pass the 2 mm and 2% criteria, and the point dose measurement must be within 2% of the calculated dose. All low dose region/critical structure point dose measurements were evaluated on a patient by patient basis. The criteria we recommend can be useful for the routine evaluation of treatment plans for tomotherapy systems. PMID- 16475780 TI - On using an adaptive neural network to predict lung tumor motion during respiration for radiotherapy applications. AB - In this study we address the problem of predicting the position of a moving lung tumor during respiration on the basis of external breathing signals--a technique used for beam gating, tracking, and other dynamic motion management techniques in radiation therapy. We demonstrate the use of neural network filters to correlate tumor position with external surrogate markers while simultaneously predicting the motion ahead in time, for situations in which neither the breathing pattern nor the correlation between moving anatomical elements is constant in time. One pancreatic cancer patient and two lung cancer patients with mid/upper lobe tumors were fluoroscopically imaged to observe tumor motion synchronously with the movement of external chest markers during free breathing. The external marker position was provided as input to a feed-forward neural network that correlated the marker and tumor movement to predict the tumor position up to 800 ms in advance. The predicted tumor position was compared to its observed position to establish the accuracy with which the filter could dynamically track tumor motion under nonstationary conditions. These results were compared to simplified linear versions of the filter. The two lung cancer patients exhibited complex respiratory behavior in which the correlation between surrogate marker and tumor position changed with each cycle of breathing. By automatically and continuously adjusting its parameters to the observations, the neural network achieved better tracking accuracy than the fixed and adaptive linear filters. Variability and instability in human respiration complicate the task of predicting tumor position from surrogate breathing signals. Our results show that adaptive signal processing filters can provide more accurate tumor position estimates than simpler stationary filters when presented with nonstationary breathing motion. PMID- 16475782 TI - A global bioheat model with self-tuning optimal regulation of body temperature using Hebbian feedback covariance learning. AB - In the lower brain, body temperature is continually being regulated almost flawlessly despite huge fluctuations in ambient and physiological conditions that constantly threaten the well-being of the body. The underlying control problem defining thermal homeostasis is one of great enormity: Many systems and sub systems are involved in temperature regulation and physiological processes are intrinsically complex and intertwined. Thus the defining control system has to take into account the complications of nonlinearities, system uncertainties, delayed feedback loops as well as internal and external disturbances. In this paper, we propose a self-tuning adaptive thermal controller based upon Hebbian feedback covariance learning where the system is to be regulated continually to best suit its environment. This hypothesis is supported in part by postulations of the presence of adaptive optimization behavior in biological systems of certain organisms which face limited resources vital for survival. We demonstrate the use of Hebbian feedback covariance learning as a possible self-adaptive controller in body temperature regulation. The model postulates an important role of Hebbian covariance adaptation as a means of reinforcement learning in the thermal controller. The passive system is based on a simplified 2-node core and shell representation of the body, where global responses are captured. Model predictions are consistent with observed thermoregulatory responses to conditions of exercise and rest, and heat and cold stress. An important implication of the model is that optimal physiological behaviors arising from self-tuning adaptive regulation in the thermal controller may be responsible for the departure from homeostasis in abnormal states, e.g., fever. This was previously unexplained using the conventional "set-point" control theory. PMID- 16475781 TI - Magnetic confinement of electron and photon radiotherapy dose: a Monte Carlo simulation with a nonuniform longitudinal magnetic field. AB - It recently has been shown experimentally that the focusing provided by a longitudinal nonuniform high magnetic field can significantly improve electron beam dose profiles. This could permit precise targeting of tumors near critical areas and minimize the radiation dose to surrounding healthy tissue. The experimental results together with Monte Carlo simulations suggest that the magnetic confinement of electron radiotherapy beams may provide an alternative to proton or heavy ion radiation therapy in some cases. In the present work, the external magnetic field capability of the Monte Carlo code PENELOPE was utilized by providing a subroutine that modeled the actual field produced by the solenoid magnet used in the experimental studies. The magnetic field in our simulation covered the region from the vacuum exit window to the phantom including surrounding air. In a longitudinal nonuniform magnetic field, it is observed that the electron dose can be focused in both the transverse and longitudinal directions. The measured dose profiles of the electron beam are generally reproduced in the Monte Carlo simulations to within a few percent in the region of interest provided that the geometry and the energy of the incident electron beam are accurately known. Comparisons for the photon beam dose profiles with and without the magnetic field are also made. The experimental results are qualitatively reproduced in the simulation. Our simulation shows that the excessive dose at the beam entrance is due to the magnetic field trapping and focusing scattered secondary electrons that were produced in the air by the incident photon beam. The simulations also show that the electron dose profile can be manipulated by the appropriate control of the beam energy together with the strength and displacement of the longitudinal magnetic field. PMID- 16475783 TI - A dosimetric comparison of 169Yb versus 192Ir for HDR prostate brachytherapy. AB - For the purpose of evaluating the use of 169Yb for prostate High Dose Rate brachytherapy (HDR), a hypothetical 169Yb source is assumed with the exact same design of the new microSelectron source replacing the 192Ir active core by pure 169Yb metal. Monte Carlo simulation is employed for the full dosimetric characterization of both sources and results are compared following the AAPM TG 43 dosimetric formalism. Monte Carlo calculated dosimetry results are incorporated in a commercially available treatment planning system (SWIFT), which features an inverse treatment planning option based on a multiobjective dose optimization engine. The quality of prostate HDR brachytherapy using the real 192Ir and hypothetical 169Yb source is compared in a comprehensive analysis of different prostate implants in terms of the multiobjective dose optimization solutions as well as treatment quality indices such as Dose Volume Histograms (DVH) and the Conformal Index (COIN). Given that scattering overcompensates for absorption in intermediate photon energies and distances in the range of interest to prostate HDR brachytherapy, 169Yb proves at least equivalent to 192Ir irrespective of prostate volume. This has to be evaluated in view of the shielding requirements for the 169Yb energies that are minimal relative to that for 192Ir. PMID- 16475784 TI - Microdosimetric evaluation of the 400 MeV/nucleon carbon beam at HIMAC. AB - Microdosimetric single event spectra were determined as a function of depth in an acrylic phantom for the carbon beam at HIMAC using a tissue equivalent proportional counter (TEPC) coupled to a scintillation counter system. The fragments produced by the carbon beam were identified by the deltaE-time of flight distribution obtained from two scintillation counters which were positioned at the up- and down-stream of the TEPC. Lineal energy distribution for the carbon beam and its five fragments, namely, proton, helium, lithium, beryllium, and boron ions, were measured in the lineal-energy range of 5-1000 keV/microm at five phantom depths between 0 and 230 mm. The dose distribution for the carbon beam and its fragments were obtained separately. The relative biological effectiveness (RBE) of the carbon beam in the phantom was calculated using a response function. The maximum RBE for the carbon beam was found to be about 5 near the Bragg peak. It was observed to rapidly decrease for Bragg peaks occurring at deeper positions in the phantom. The dose from the beam fragments accounted for about 30% to the total dose, however, its contribution to the RBE was less than 17%. PMID- 16475785 TI - 2-Step IMAT and 2-Step IMRT in three dimensions. AB - In two dimensions, 2-Step Intensity Modulated Arc Therapy (2-Step IMAT) and 2 Step Intensity Modulated Radiation Therapy (IMRT) were shown to be powerful methods for the optimization of plans with organs at risk (OAR) (partially) surrounded by a target volume (PTV). In three dimensions, some additional boundary conditions have to be considered to establish 2-Step IMAT as an optimization method. A further aim was to create rules for ad hoc adaptations of an IMRT plan to a daily changing PTV-OAR constellation. As a test model, a cylindrically symmetric PTV-OAR combination was used. The centrally placed OAR can adapt arbitrary diameters with different gap widths toward the PTV. Along the rotation axis the OAR diameter can vary, the OAR can even vanish at some axis positions, leaving a circular PTV. The width and weight of the second segment were the free parameters to optimize. The objective function f to minimize was the root of the integral of the squared difference of the dose in the target volume and a reference dose. For the problem, two local minima exist. Therefore, as a secondary criteria, the magnitude of hot and cold spots were taken into account. As a result, the solution with a larger segment width was recommended. From plane to plane for varying radii of PTV and OAR and for different gaps between them, different sets of weights and widths were optimal. Because only one weight for one segment shall be used for all planes (respectively leaf pairs), a strategy for complex three-dimensional (3-D) cases was established to choose a global weight. In a second step, a suitable segment width was chosen, minimizing f for this global weight. The concept was demonstrated in a planning study for a cylindrically symmetric example with a large range of different radii of an OAR along the patient axis. The method is discussed for some classes of tumor/organ at risk combinations. Noncylindrically symmetric cases were treated exemplarily. The product of width and weight of the additional segment as well as the integral across the segment profile was demonstrated to be an important value. This product was up to a factor of 3 larger than in the 2-D case. Even in three dimensions, the optimized 2-Step IMAT increased the homogeneity of the dose distribution in the PTV profoundly. Rules for adaptation to varying target-OAR combinations were deduced. It can be concluded that 2-Step IMAT and 2-Step IMRT are also applicable in three dimensions. In the majority of cases, weights between 0.5 and 2 will occur for the additional segment. The width-weight product of the second segment is always smaller than the normalized radius of the OAR. The width-weight product of the additional segment is strictly connected to the relevant diameter of the organ at risk and the target volume. The derived formulas can be helpful to adapt an IMRT plan to altering target shapes. PMID- 16475786 TI - 198Au seeds. PMID- 16475787 TI - Prostaglandin F2alpha formation from prostaglandin H2 by prostaglandin F synthase (PGFS): crystal structure of PGFS containing bimatoprost. AB - Prostaglandin H(2) (PGH(2)) formed from arachidonic acid is an unstable intermediate and is efficiently converted into more stable arachidonate metabolites by the action of enzymes. Prostaglandin F synthase (PGFS) has dual catalytic activities: formation of PGF(2)(alpha) from PGH(2) by the PGH(2) 9,11 endoperoxide reductase activity and 9alpha,11beta-PGF(2) (PGF(2)(alphabeta)) from PGD(2) by the PGD(2) 11-ketoreductase activity in the presence of NADPH. Bimatoprost (BMP), which is a highly effective ocular hypotensive agent, is a PGF(2)(alpha) analogue that inhibits both the PGD(2) 11-ketoreductase and PGH(2) 9,11-endoperoxide reductase activities of PGFS. To examine the catalytic mechanism of PGH(2) 9,11-endoperoxide reductase, a crystal structure of PGFS[NADPH + BMP] has been determined at 2.0 A resolution. BMP binds near the PGD(2) binding site, but the alpha- and omega-chains of BMP are locate on the omega- and alpha-chains of PGD(2), respectively. Consequently, the bound BMP and PGD(2) direct their opposite faces of the cyclopentane moieties toward the nicotinamide ring of the bound NADP. The alpha- and omega-chains of BMP are involved in H-bonding with protein residues, while the cyclopentane moiety is surrounded by water molecules and is not directly attached to either the protein or the bound NADPH, indicating that the cyclopentane moiety is movable in the active site. From the complex structure, two model structures of PGFS containing PGF(2)(alpha) and PGH(2) were built. On the basis of the model structures and inhibition data, a putative catalytic mechanism of PGH(2) 9,11-endoperoxide reductase of PGFS is proposed. Formation of PGF(2)(alpha) from PGH(2) most likely involves a direct hydride transfer from the bound NADPH to the endoperoxide of PGH(2) without the participation of specific amino acid residues. PMID- 16475788 TI - Annotating enzymes of unknown function: N-formimino-L-glutamate deiminase is a member of the amidohydrolase superfamily. AB - The functional assignment of enzymes that catalyze unknown chemical transformations is a difficult problem. The protein Pa5106 from Pseudomonas aeruginosa has been identified as a member of the amidohydrolase superfamily by a comprehensive amino acid sequence comparison with structurally authenticated members of this superfamily. The function of Pa5106 has been annotated as a probablechlorohydrolase or cytosine deaminase. A close examination of the genomic content of P. aeruginosa reveals that the gene for this protein is in close proximity to genes included in the histidine degradation pathway. The first three steps for the degradation of histidine include the action of HutH, HutU, and HutI to convert L-histidine to N-formimino-L-glutamate. The degradation of N-formimino L-glutamate to L-glutamate can occur by three different pathways. Three proteins in P. aeruginosa have been identified that catalyze two of the three possible pathways for the degradation of N-formimino-L-glutamate. The protein Pa5106 was shown to catalyze the deimination of N-formimino-L-glutamate to ammonia and N formyl-L-glutamate, while Pa5091 catalyzed the hydrolysis of N-formyl-L-glutamate to formate and L-glutamate. The protein Pa3175 is dislocated from the hut operon and was shown to catalyze the hydrolysis of N-formimino-L-glutamate to formamide and L-glutamate. The reason for the coexistence of two alternative pathways for the degradation of N-formimino-L-glutamate in P. aeruginosa is unknown. PMID- 16475789 TI - Formation of virus-like clusters is an intrinsic property of the tumor necrosis factor family member BAFF (B cell activating factor). AB - The oligomeric state of BAFF (B cell activing factor), a tumor necrosis factor (TNF) family cytokine that plays a critical role in B cell development and survival, has been the subject of recent debate. Myc-tagged BAFF starting at residue Gln136 was previously reported to crystallize as trimers at pH 4.5, whereas a histidine-tagged construct of BAFF, starting at residue Ala134, formed a virus-like cluster containing 60 monomers when crystallized at pH 9.0. The formation of the BAFF 60-mer was pH dependent, requiring pH >or= 7.0. More recently, 60-mer formation was suggested to be artificially induced by the histidine tag, and it was proposed that BAFF, like all other TNF family members, is trimeric. We report here that a construct of BAFF with no amino-terminal tag (Ala134-BAFF) can form a 60-mer in solution. Using size exclusion chromatography and static light scattering to monitor trimer to 60-mer ratios in BAFF preparations, we find that 60-mer formation is pH-dependent and requires histidine 218 within the DE loop of BAFF. Biacore measurements established that the affinity of Ala134-BAFF for the BAFF receptor BAFFR/BR3 is similar to that of myc-Gln136-BAFF, which is exclusively trimeric in solution. However, Ala134-BAFF is more efficacious than myc-Gln136-BAFF in inducing B cell proliferation in vitro. We additionally show that BAFF that is processed and secreted by 293T cells transfected with full-length BAFF, or by a histiocytic lymphoma cell line (U937) that expresses BAFF endogenously, forms a pH-dependent 60-mer in solution. Our results indicate that the formation of the 60-mer in solution by the BAFF extracellular domain is an intrinsic property of the protein, and therefore that this more active form of BAFF may be physiologically relevant. PMID- 16475790 TI - Molecular mechanisms and binding site location for the noncompetitive antagonist crystal violet on nicotinic acetylcholine receptors. AB - We investigated the molecular mechanisms and the binding site location for the fluorophor crystal violet (CrV), a noncompetitive antagonist of the nicotinic acetylcholine receptor (AChR). To this end, radiolabeled competition binding, fluorescence spectroscopy, Schild-type analysis, patch-clamp recordings, and molecular dynamics approaches were used. The results indicate that (i) CrV interacts with the desensitized Torpedo AChR with higher affinity than with the resting state at several temperatures (5-37 degrees C); (ii) CrV-induced inhibition of the phencyclidine (PCP) analogue [(3)H]thienylcyclohexylpiperidine binding to the desensitized or resting AChR is mediated by a steric mechanism; (iii) tetracaine inhibits CrV binding to the resting AChR, probably by a steric mechanism; (iv) barbiturates modulate CrV binding to the resting AChR by an allosteric mechanism; (v) CrV itself induces AChR desensitization; (vi) CrV decreases the peak of macroscopic currents by acting on the resting AChR but without affecting the desensitization rate from the open state; and (vii) two tertiary amino groups from CrV may bind to the alpha1-Glu(262) residues (located at position 20') in the resting state. We conclude that the CrV binding site overlaps the PCP locus in the resting and desensitized state. The noncompetitive action of CrV may be explained by an allosteric mechanism in which the binding of CrV to the extracellular mouth of the resting receptor leads to an inhibition of channel opening. Binding of CrV probably increases desensitization of the resting channel and stabilizes the desensitized state. PMID- 16475791 TI - The mid-region of parathyroid hormone (1-34) serves as a functional docking domain in receptor activation. AB - Elucidating the bimolecular interface between parathyroid hormone (PTH) and its cognate G protein-coupled receptor (PTHR1) should yield insights into the basis of molecular recognition and the mechanism of ligand-mediated intracellular signaling for a system that is critically important in regulating calcium levels in blood. We used photoaffinity scanning (PAS) to identify key ligand-receptor interactions for residues from the unstructured mid-region domain of PTH-(1-34). Four PTH analogues, containing a single photoreactive p-benzoylphenylalanine (Bpa) residue in position 11, 15, 18, or 21, were found to photo-cross-link within receptor regions [165-176], [183-189], [190-298], and [165-176], respectively. Addition of these mid-region contacts as constraints to our previously proposed model of the PTH-PTHR1 complex and extensive molecular simulation experiments enables substantial refinement of the model. Specifically, (1) the overall receptor-bound conformation of the hormone is not extended, but bent; (2) helix [169-176] of the N-terminal extracellular domain (N-ECD) of the receptor is redirected toward the heptahelical bundle; and (3) the hormone traverses between the top of transmembrane (TM) helices 1 and 2, rather than between TM-7 and TM-1. This significantly alters the model of both the receptor bound tertiary structure of the hormone and the topological orientation of the C terminus of the N-ECD in the hormone-receptor bimolecular complex. We propose that the mid-region of PTH-(1-34) has a role in fixing, by extensive contacts with the receptor, the entry of the N-terminal helix of the hormone into the heptahelical bundle between TM-1 and TM-2. This anchorage would orient the amino terminus into position to activate the receptor. PMID- 16475792 TI - Zn-alpha2-glycoprotein, an MHC class I-related glycoprotein regulator of adipose tissues: modification or abrogation of ligand binding by site-directed mutagenesis. AB - Zn-alpha(2)-glycoprotein (ZAG) is a soluble lipid-mobilizing factor associated with cancer cachexia and is a novel adipokine. Its X-ray crystal structure reveals a poly(ethylene glycol) molecule, presumably substituting for a higher affinity natural ligand, occupying an apolar groove between its alpha(1) and alpha(2) domain helices that corresponds to the peptide binding groove in class I MHC proteins. We previously provided evidence that the groove is a binding site for hydrophobic ligands that may relate to the protein's signaling function and that the natural ligands are probably (polyunsaturated) fatty acid-like. Using fluorescence-based binding assays and site-directed mutagenesis, we now demonstrate formally that the groove is indeed the binding site for hydrophobic ligands. We also identify amino acid positions that are involved in ligand binding and those that control the shape and exposure to solvent of the binding site itself. Some of the mutants showed minimal effects on their binding potential, one showed enhanced binding, and several were completely nonbinding. Particularly notable is Arg-73, which projects into one end of the binding groove and is the sole charged amino acid adjacent to the ligand. Replacing this amino acid with alanine abolished ligand binding and closed the groove to solvent. Arg 73 may therefore have an unexpected dual role in binding site access and anchor for an amphiphilic ligand. These data add weight to the distinctiveness of ZAG among MHC class I-like proteins in addition to providing defined binding-altered mutants for cellular signaling studies and potential medical applications. PMID- 16475793 TI - Binding of laminin alpha1-chain LG4-5 domain to alpha-dystroglycan causes tyrosine phosphorylation of syntrophin to initiate Rac1 signaling. AB - Previously, a signaling pathway was described [Oak, Zhou, and Jarrett (2003) J. Biol. Chem. 278, 39287-39295] that links matrix laminin binding on the outside of the sarcolemma to Grb2 binding to syntrophin on the inside surface of the sarcolemma and by way of Grb2-Sos1-Rac1-PAK1-JNK ultimately results in the phosphorylation of c-jun on Ser(65). How this signaling is initiated was investigated. Grb2-binding to syntrophin is increased by the addition of either laminin-1 or the isolated laminin alpha1 globular domain modules LG4-5, a protein referred to as E3. This identifies the LG4-5 sequences as the region of laminin responsible for signaling. Since laminin alpha1 LG4 is known to bind alpha dystroglycan, this directly implicates alpha-dystroglycan as the laminin signaling receptor. E3 or laminin-1 increase Grb2-binding and Rac1 activation. In the presence of E3 or laminin-1, syntrophin is phosphorylated on a tyrosine residue, and this increases and alters Grb2 binding. The alpha-dystroglycan antibody, IIH6, which blocks binding of laminins to alpha-dystroglycan, blocks both the laminin-induced Sos1/2 recruitment and syntrophin phosphorylation, showing that it is alpha-dystroglycan binding the LG4-5 region of laminin that is responsible. The C-terminal SH3 domain of Grb2 (C-SH3) binds only to nonphosphorylated syntrophin, and phosphorylation causes the Grb2 SH2 domain to bind and prevents SH3 binding. Syntrophin, tyrosine phosphate, beta-dystroglycan, and Rac1 all co-localize to the sarcolemma of rat muscle sections. A model for how this phosphorylation may initiate downstream events in laminin signaling is presented. PMID- 16475794 TI - Triplet-state conformational changes in 15-cis-spheroidene bound to the reaction center from Rhodobacter sphaeroides 2.4.1 as revealed by time-resolved EPR spectroscopy: strengthened hypothetical mechanism of triplet-energy dissipation. AB - Time-resolved EPR spectra of 15-cis-spheroidene bound to the reaction center from Rhodobacter sphaeroides 2.4.1 were recorded at low temperatures. (1) A three component analysis of the spectral-data matrices by singular-value decomposition followed by global fitting identified the transformation of the triplet carotenoid, (3)Car(I) --> (3)Car(II); during this process, the leak of the triplet population was suggested. A four-component analysis suggested the presence of a representative intermediate, (3)Car(R), that forms a leak channel of the triplet population. (2) A theoretical calculation of the zero-field splitting parameters, |D| and |E|, by the use of a polyene model, showed that the transformation, (3)Car(I) --> (3)Car(R) --> (3)Car(II), accompanies the conformational changes of (0 degrees , 0 degrees , 0 degrees ) --> (+20 degrees , -20 degrees , +20 degrees ) --> (+45 degrees , -40 degrees , +40 degrees ) around the central cis C15=C15', trans C13=C14, and trans C11=C12 bonds, respectively. (3) The initial, rapid decrease followed by the inversion of spin polarization along the z axis of (3)Car was observed, which was correlated with a change in the spin angular momentum. (4) In reference to the binding pocket of the Car, determined by X-ray crystallography, the conformational changes were ascribed to the intrinsic isomerization property of 15-cis (3)Car as well as the Car-peptide intermolecular interaction; a detailed picture was proposed. All of the above results support the mechanism of triplet-energy dissipation proposed previously: the rotational motions around the central double bonds cause a change in the orbital angular momentum and, through the spin-orbit coupling, a change in the spin angular momentum, which enhances the T(1) --> S(0) intersystem crossing dissipating the triplet energy. PMID- 16475795 TI - Energetics of a possible proton exit pathway for water oxidation in photosystem II. AB - The crystal structure of photosystem II (PSII) at 3.0-A resolution suggests that titratable residues on the lumenal side of D1/D2 and PsbO form a polar channel, which might serve as a proton exit pathway associated with water oxidation on the Mn-cluster. With full account of protein environment, we calculated the pK(a) of these residues by solving the linearized Poisson-Boltzmann equation. Along the prospective proton channel, the calculated pK(a) of titratable residues (namely via D1-Asp61, D1-Glu65, D2-Glu312, D2-Lys317 D1-Asp59, D1-Arg64, PsbO-Arg152, and PsbO-Asp224) monotonically increase from the Mn-cluster to the lumenal bulk side. We suggest that these residues form the exit pathway guiding protons, which are released at the Mn-cluster as a product of water oxidation, in an exergonic process out of PSII. Upon the S2 to S3 transition, CP43-Arg357 showed a dramatic deprotonation of ca. one H(+), suggesting that this residue is coupled to the redox states of the Mn-cluster and the tyrosine Y(Z). The calculated pK(a) values of 4.2-4.4 for D2-Glu312 and those of approximately 8-10.9 for D1-Asp59 and D1 Arg64 are indicative of the experimentally determined pK(a) values for inhibition of S-state transitions. Upon removal of the atomic coordinates of PsbO, the pK(a) of these residues are dramatically affected, indicating a significant role of PsbO in tuning the pK(a) of those residues in the proton exit pathway. PMID- 16475796 TI - Functional implications of the propionate 7-arginine 220 interaction in the FixLH oxygen sensor from Bradyrhizobium japonicum. AB - BjFixL from Bradyrhizobium japonicum is a heme-based oxygen sensor implicated in the signaling cascade that enables the bacterium to adapt to fluctuating oxygen levels. Signal transduction is initiated by the binding of O(2) to the heme domain of BjFixL, resulting in protein conformational changes that are transmitted to a histidine kinase domain. We report structural changes of the heme and its binding pocket in the Fe(II) deoxy and Fe(III) met states of the wild-type BjFixLH oxygen sensor domain and four mutants of the highly conserved residue arginine 220. UV-visible, electron paramagnetic resonance, and resonance Raman spectroscopies all showed that the heme iron of the R220H mutant is unexpectedly six-coordinated at physiological pH in the Fe(III) state but undergoes pH- and redox-dependent coordination changes. This behavior is unprecedented for FixL proteins, but is reminiscent of another oxygen sensor from E. coli, EcDos. All mutants in their deoxy states are five-coordinated Fe(II), although we report rupture of the residue 220-propionate 7 interaction and structural modifications of the heme conformation as well as propionate geometry and flexibility. In this work, we conclude that part of the structural reorganization usually attributed to O(2) binding in the wild-type protein is in fact due to rupture of the Arg220-P7 interaction. Moreover, we correlate the structural modifications of the deoxy Fe(II) states with k(on) values and conclude that the Arg220-P7 interaction is responsible for the lower O(2) and CO k(on) values reported for the wild-type protein. PMID- 16475797 TI - Selectivity of tryptophan residues in mediating photolysis of disulfide bridges in goat alpha-lactalbumin. AB - Goat alpha-lactalbumin (GLA) contains four tryptophan (Trp) residues and four disulfide bonds. Illumination with near-UV light results in the cleavage of disulfide bridges and in the formation of free thiols. To obtain information about the reaction products, the illuminated protein was carbamidomethylated and digested with trypsin and the peptides were analyzed by mass spectrometry. Peptides containing Cys120Cam, Cys61Cam, or Cys91Cam were detected, as well as two peptides containing a new Cys-Lys cross-link. In one, Cys6 was cross-linked to Lys122, while the cross-link in the second was either a Cys91-Lys79 or Cys73 Lys93 cross-link; however, the exact linkage could not be defined. The results demonstrate photolytic cleavage of the Cys6-Cys120, Cys61-Cys77, and Cys73-Cys91 disulfide bonds. While photolysis of Cys6-Cys120 and Cys73-Cys91 disulfide bonds in GLA has been reported, cleavage of the Cys61-Cys77 disulfide bonds has not been previously detected. To examine the contribution of the individual Trp residues, we constructed the GLA mutants, W26F, W60F, W104F, and W118F, by replacing single Trp residues with phenylalanine (Phe). The substitution of each Trp residue led to less thiol production compared to that for wild-type GLA, showing that each Trp residue in GLA contributed to the photolytic cleavage of disulfide bridges. The specificity was expressed by the nature of the reaction products. No cleavage of the Cys6-Cys120 disulfide bridge was detected when the W26F mutant was illuminated, and no cleavage of the Cys73-Cys91 disulfide bridge was seen following illumination of W26F or W104F. In contrast, Cys61Cam, resulting from the cleavage of the Cys61-Cys77 disulfide bridge, was found following illumination of any of the mutants. PMID- 16475798 TI - Quantitative assessment of intrinsic carbonic anhydrase activity and the capacity for bicarbonate oxidation in photosystem II. AB - On the basis of equilibrium isotopic distribution experiments using (18)O-labeled water, it is generally accepted that water is the sole substrate for O(2) production by photosystem II (PSII). Nevertheless, recent studies indicating a direct interaction between bicarbonate and the donor side of PSII have been used to hypothesize that bicarbonate may have been a physiologically important substrate for O(2) production during the evolution of PSII [Dismukes, G. C., Klimov, V. V., Baranov, S. V., Kozlov, Y. N., DasGupta, J., and Tyryshikin, A. (2001) Proc. Natl. Acad. Sci. U.S.A. 98, 2170-2175]. To test out this hypothesis and to determine whether contemporary oxygenic organisms have the capacity to oxidize bicarbonate, we employed special rapid-mixing isotopic experiments using (18)O/(13)C-labeled bicarbonate to quantify the inherent carbonic anhydrase activity in PSII samples and the potential flux of oxygen from bicarbonate into the photosynthetically produced O(2). The measurements were made on PSII samples prepared from spinach, Thermosynechococcus elongatus, and Arthrospira maxima. For the latter organism, a strain was used that grows naturally in an alkaline, high (bi)carbonate soda lake in Africa. The results reveal that bicarbonate is not the substrate for O(2) production in these contemporary oxygenic photoautotrophs when assayed under single turnover conditions. PMID- 16475799 TI - Design of a minimal polypeptide unit for bacteriochlorophyll binding and self assembly based on photosynthetic bacterial light-harvesting proteins. AB - We introduce LH1beta24, a minimal 24 amino acid polypeptide that binds and assembles bacteriochlorophylls (BChls) in micelles of octyl beta-glucoside (OG) into complexes with spectral properties that resemble those of B820, a universal intermediate in the assembly of native purple bacterial light-harvesting complexes (LHs). LH1beta24 was designed by a survey of sequences and crystal structures of bacterial LH proteins from different organisms combined with currently available information from in vitro reconstitution studies and genetically modified LHs in vivo. We took as a template for the design sphbeta31, a truncated 31 amino acid analogue of the native beta-apoprotein from the core LH complex of Rhodobacter sphaeroides. This peptide self-assembles with BChls to form B820 and, upon cooling and lowering OG concentration, forms red-shifted B850 spectral species that are considered analogous to native LH complexes. We find that LH1beta24 self-assembles with BChl in OG to form homodimeric B820-type subunits comprising two LH1beta24 and two BChl molecules per subunit. We demonstrate, by modeling the structure using the highly homologous structure of LH2 from Rhodospirillum molischianum, that it has the minimal size for BChl binding. Additionally, we have compared the self-assembly of sphbeta31 and LH1beta24 with BChls and discovered that the association enthalpies and entropies of both species are similar to those measured for native LH1 from Rhodospirillum rubrum. However, sphbeta31 readily aggregates into intermediate higher oligomeric species and further to form B850 species; moreover, the assembly process of these oligomers is not reversible, and they are apparently large nonspecific BChl peptide coaggregates rather than well-defined nativelike LH complexes. Similar aggregates were observed during LH1beta24 assembly, but these were formed less readily and required lower temperatures than sphbeta31. In view of these results, we reevaluate previous in vitro reconstitution studies and propose alternative templates for new designs. PMID- 16475800 TI - Enzyme:substrate hydrogen bond shortening during the acylation phase of serine protease catalysis. AB - Atomic resolution (4)-glycosidic bond in the bacterial cell wall heteropolymer peptidoglycan between the N-acetylmuramic acid (MurNAc) and N-acetylglucosamine (GlcNAc) residues with the concomitant formation of a 1,6 anhydromuramoyl residue. On the basis of both sequence alignments with and structural considerations of soluble lytic transglycosylase Slt35 from Escherichia coli, four residues were predicted to be involved in substrate binding at the -1 subsite in the soluble derivative of Pseudomonas aeruginosa membrane-bound lytic transglycosylase MltB. These residues were targeted for site specific replacement, and the effect on substrate binding and catalysis was determined. The residues Arg187 and Arg188, believed to be involved in binding the stem peptide on MurNAc, were shown to play an important role in substrate binding, as evidenced by peptidoglycan affinity assays and SUPREX analysis using MurNAc-dipeptide as ligand. The Michaelis-Menten parameters were determined for the respective mutants using insoluble peptidoglycan as substrate. In addition to affecting the steady-state binding of ligand to enzyme, as indicated by increases in K(M) values, significant decreases in k(cat) values suggested that replacement of either Arg187 and Arg188 with alanine perturbed the stabilization of both the transition state(s) and reaction intermediate. Thus, it appears that Arg187 and Arg188 are vital for proper orientation of the substrate in the active site, and furthermore this supports the proposed role of the stem peptide at binding subsite -2 in catalysis. Replacement of Gln100, a residue that would appear to interact with the N-acetyl group on MurNAc, did not show any changes in substrate affinity or activity. PMID- 16475803 TI - Cytidine 5'-monophosphate (CMP)-induced structural changes in a multifunctional sialyltransferase from Pasteurella multocida. AB - Sialyltransferases catalyze reactions that transfer a sialic acid from CMP-sialic acid to an acceptor (a structure terminated with galactose, N acetylgalactosamine, or sialic acid). They are key enzymes that catalyze the synthesis of sialic acid-containing oligosaccharides, polysaccharides, and glycoconjugates that play pivotal roles in many critical physiological and pathological processes. The structures of a truncated multifunctional Pasteurella multocida sialyltransferase (Delta24PmST1), in the absence and presence of CMP, have been determined by X-ray crystallography at 1.65 and 2.0 A resolutions, respectively. The Delta24PmST1 exists as a monomer in solution and in crystals. Different from the reported crystal structure of a bifunctional sialyltransferase CstII that has only one Rossmann domain, the overall structure of the Delta24PmST1 consists of two separate Rossmann nucleotide-binding domains. The Delta24PmST1 structure, thus, represents the first sialyltransferase structure that belongs to the glycosyltransferase-B (GT-B) structural group. Unlike all other known GT-B structures, however, there is no C-terminal extension that interacts with the N-terminal domain in the Delta24PmST1 structure. The CMP binding site is located in the deep cleft between the two Rossmann domains. Nevertheless, the CMP only forms interactions with residues in the C-terminal domain. The binding of CMP to the protein causes a large closure movement of the N-terminal Rossmann domain toward the C-terminal nucleotide-binding domain. Ser 143 of the N-terminal domain moves up to hydrogen-bond to Tyr 388 of the C terminal domain. Both Ser 143 and Tyr 388 form hydrogen bonds to a water molecule, which in turn hydrogen-bonds to the terminal phosphate oxygen of CMP. These interactions may trigger the closure between the two domains. Additionally, a short helix near the active site seen in the apo structure becomes disordered upon binding to CMP. This helix may swing down upon binding to donor CMP-sialic acid to form the binding pocket for an acceptor. PMID- 16475804 TI - The G473D mutation impairs dimerization and catalysis in human sulfite oxidase. AB - Among the mutations identified in patients with isolated sulfite oxidase deficiency, the G473D variant is of particular interest since sedimentation analysis reveals that this variant is a monomer, and the importance of the wild type dimeric state of mammalian sulfite oxidase is not yet well understood. Analysis of recombinant G473D sulfite oxidase indicated that it is severely impaired both in the ability to bind sulfite and in catalysis, with a second order rate constant 5 orders of magnitude lower than that of the wild type. To elucidate the specific reasons for the severe effects seen in the G473D variant, several other variants were created, including G473A, G473W, and the double mutant R212A/G473D. Despite the inability to form a stable dimer, the G473W variant had 5-fold higher activity than G473D and nearly wild-type activity at pH 7.0 when ferricyanide was the electron acceptor. In contrast, the R212A/G473D variant demonstrated some ability to oligomerize but had undetectable activity. The G473A variant retained the ability to dimerize and had steady-state activity that was comparable to that of the wild type. Furthermore, stopped-flow analysis of the reductive half-reaction of this variant yielded a rate constant nearly 3 times higher than that of the wild type. Examination of the secondary structures of the variants by CD spectroscopy indicated significant random-coil formation in G473D, G473W, and R212A/G473D. These results demonstrate that both the charge and the large size of an Asp residue in this position contribute to the severe effects seen in a patient with the G473D mutation, by causing partial misfolding and monomerization of sulfite oxidase and attenuating both substrate binding and catalytic efficiency during the reaction cycle. PMID- 16475806 TI - The 108M polymorph of human catechol O-methyltransferase is prone to deformation at physiological temperatures. AB - The human gene for catechol O-methyltransferase (COMT) contains a common polymorphism that results in substitution of methionine (M) for valine (V) at residue 108 of the soluble form of the protein. While the two proteins have similar kinetic properties, 108M COMT loses activity more rapidly than 108V COMT at 37 degrees C. The cosubstrate S-adenosylmethionine (SAM) stabilizes the activity of 108M COMT at 40 degrees C. The 108M allele has been associated with increased risk for breast cancer, obsessive-compulsive disorder, and aggressive and highly antisocial manifestations of schizophrenia. In the current work, we have constructed homology models for both human COMT polymorphs and performed molecular dynamics simulations of these models at 25, 37, and 50 degrees C to explore the structural consequences of the 108V/M polymorphism. The simulations indicated that replacing valine with the larger methionine residue led to greater solvent exposure of residue 108 and heightened packing interactions between M108 and helices alpha2, alpha4 (especially with R78), and alpha5. These altered packing interactions propagated subtle changes between the polymorphic site and the active site 16 A away, leading to a loosening of the active site. At physiological temperature, 108M COMT sampled a larger distribution of conformations than 108V. 108M COMT was more prone to active-site distortion and had greater overall, and SAM binding site, solvent accessibility than 108V COMT at 37 degrees C. Similar structural perturbations were observed in the 108V protein only at 50 degrees C. Addition of SAM tightened up the cosubstrate pocket in both proteins and prevented the altered packing at the polymorphic site in 108M COMT. PMID- 16475808 TI - Ras regulation by reactive oxygen and nitrogen species. AB - Ras GTPases cycle between inactive GDP-bound and active GTP-bound states to modulate a diverse array of processes involved in cellular growth control. We have previously shown that both NO/O(2) (via nitrogen dioxide, (*)NO(2)) and superoxide radical anion (O(2)(*)(-)) promote Ras guanine nucleotide dissociation. We now show that hydrogen peroxide in the presence of transition metals (i.e., H(2)O(2)/transition metals) and peroxynitrite also trigger radical based Ras guanine nucleotide dissociation. The primary redox-active reaction species derived from H(2)O(2)/transition metals and peroxynitrite is O(2)(*)(-) and (*)NO(2), respectively. A small fraction of hydroxyl radical (OH(*)) is also present in both. We also show that both carbonate radical (CO(3)(*)(-)) and (*)NO(2), derived from the mixture of peroxynitrite and bicarbonate, facilitate Ras guanine nucleotide dissociation. We further demonstrate that NO/O(2) and O(2)(*)(-) promote Ras GDP exchange with GTP in the presence of a radical quenching agent, ascorbate, or NO, and generation of Ras-GTP promotes high affinity binding of the Ras-binding domain of Raf-1, a downstream effector of Ras. S-Nitrosylated Ras (Ras-SNO) can be formed when NO serves as a radical quenching agent, and hydroxyl radical but not (*)NO(2) or O(2)(*)(-) can further react with Ras-SNO to modulate Ras activity in vitro. However, given the lack of redox specificity associated with the high redox potential of OH(*), it is unclear whether this reaction occurs under physiological conditions. PMID- 16475807 TI - Compaction of ribosomal protein S6 by sucrose occurs only under native conditions. AB - The effect of osmolyte sucrose on the stability and compaction of the folded and unfolded states of ribosomal protein S6 from Thermus thermophilus was analyzed. Confirming previous results obtained with sodium sulfate and trehalose, refolding stopped-flow measurements of S6 show that sucrose favors the conversion of the unfolded state ensemble to a highly compact structure (75% as compact as the folded state). This conversion occurs when the unfolded state is suddenly placed under native conditions and the compact state accumulates in a transient off folding pathway. This effect of sucrose on the compaction of the unfolded state ensemble is counteracted by guanidinium hydrochloride. The compact state does not accumulate at higher guanidinium concentrations and the unfolded state ensemble does not display increased compaction in the presence of 6 M guanidinium as evaluated by collisional quenching of tryptophan fluorescence. In contrast, accessibility of the tryptophan residue of folded S6 above 1 M sucrose concentration decreased as a result of an increased compaction of the folded state. Unfolding stopped-flow measurements of S6 reflect this increased compaction of the folded state, but the unfolding pathway is not affected by sucrose. Compaction of folded and unfolded S6 induced by sucrose occurs under native conditions indicating that decreased protein conformational entropy significantly contributes to the mechanism of protein stabilization by osmolytes. PMID- 16475805 TI - Solute probes of conformational changes in open complex (RPo) formation by Escherichia coli RNA polymerase at the lambdaPR promoter: evidence for unmasking of the active site in the isomerization step and for large-scale coupled folding in the subsequent conversion to RPo. AB - Transcription initiation is a multistep process involving a series of requisite conformational changes in RNA polymerase (R) and promoter DNA (P) that create the open complex (RP(o)). Here, we use the small solutes urea and glycine betaine (GB) to probe the extent and type of surface area changes in the formation of RP(o) between Esigma(70) RNA polymerase and lambdaP(R) promoter DNA. Effects of urea quantitatively reflect changes in amide surface and are particularly well suited to detect coupled protein folding events. GB provides a qualitative probe for the exposure or burial of anionic surface. Kinetics of formation and dissociation of RP(o) reveal strikingly large effects of the solutes on the final steps of RP(o) formation: urea dramatically increases the dissociation rate constant k(d), whereas GB decreases the rate of dissociation. Formation of the first kinetically significant intermediate I(1) is disfavored in urea, and moderately favored by GB. GB slows the rate-determining step that converts I(1) to the second kinetically significant intermediate I(2); urea has no effect on this step. The most direct interpretation of these data is that recognition of promoter DNA in I(1) involves only limited conformational changes. Notably, the data support the following hypotheses: (1) the negatively charged N-terminal domain of sigma(70) remains bound in the "jaws" of polymerase in I(1); (2) the subsequent rate-determining isomerization step involves ejecting this domain from the jaws, thereby unmasking the active site; and (3) final conversion to RP(o) involves coupled folding of the mobile downstream clamp of polymerase. PMID- 16475809 TI - The L561A substitution in the nascent base-pair binding pocket of RB69 DNA polymerase reduces base discrimination. AB - Several variants of RB69 DNA polymerase (RB69 pol) with single-site replacements in the nascent base-pair binding pocket are less discriminating with respect to noncomplementary dNMP incorporation than the wild-type enzyme. To quantify the loss in base selectivity, we determined the transient-state kinetic parameters for incorporation of correct and all combinations of incorrect dNMPs by the exonuclease-deficient form of one of these RB69 pol variants, L561A, using rapid chemical quench assays. The L561A variant did not significantly alter the k(pol) and K(D) values for incorporation of correct dNMPs, but it showed increased incorporation efficiency (k(pol)/K(D)) for mispaired bases relative to the wild type enzyme. The incorporation efficiency for mispaired bases by the L561A variant ranged from 1.5 x 10(-)(5) microM(-)(1) s(-)(1) for dCMP opposite templating C to 2 x 10(-)(3) microM(-)(1) s(-)(1) for dAMP opposite templating C. These k(pol)/K(D) values are 3-60-fold greater than those observed with the wild type enzyme. The effect of the L561A replacement on the mutation frequency in vivo was determined by infecting Escherichia coli harboring a plasmid encoding the L561A variant of RB69 pol with T4 phage bearing a mutant rII locus, and the rates of reversions to rII(+) were scored. The exonuclease-proficient RB69 pol L561A displayed a weak mutator phenotype. In contrast, no progeny phage were produced after infection of E. coli, expressing an exonuclease-deficient RB69 pol L561A, with either mutant or wild-type T4 phage. This dominant-lethal phenotype was attributed to error catastrophe caused by the high rate of mutation expected from combining the pol L561A and exo(-) mutator activities. PMID- 16475810 TI - NMR structure and molecular dynamics of the in-plane membrane anchor of nonstructural protein 5A from bovine viral diarrhea virus. AB - Hepatitis C virus (HCV) nonstructural protein 5A (NS5A) is a monotopic membrane protein anchored to the membrane by an N-terminal in-plane amphipathic alpha helix. This membrane anchor is essential for the assembly of a functional viral replication complex. Although amino acid sequences differ considerably, putative membrane anchors with amphipathic features were predicted in NS5A from related Flaviviridae family members, in particular bovine viral diarrhea virus (BVDV), the prototype representative of the genus Pestivirus. We report here the NMR structure of the membrane anchor 1-28 of NS5A from BVDV in the presence of different membrane mimetic media. This anchor includes a long amphipathic alpha helix of 21 residues interacting in-plane with the membrane interface and including a putative flexible region. Molecular dynamic simulation at a water dodecane interface used to mimic the surface separating a lipid bilayer and an aqueous medium demonstrated the stability of the helix orientation and the location at the hydrophobic-hydrophilic interface. The flexible region of the helix appears to be required to allow the most favorable interaction of hydrophobic and hydrophilic side chain residues with their respective environment at the membrane interface. Despite the lack of amino acid sequence similarity, this amphipathic helix shares common structural features with that of the HCV counterpart, including a stable, hydrophobic N-terminal segment separated from the more hydrophilic C-terminal segment by a local, flexible region. These structural conservations point toward conserved roles of the N-terminal in-plane membrane anchors of NS5A in replication complex formation of HCV, BVDV, and other related viruses. PMID- 16475811 TI - Heparin accelerates gelsolin amyloidogenesis. AB - The chemical environment of the extracellular matrix may influence the tissue selective deposition observed there in gelsolin amyloid disease. Previously, we have identified the proteases that generate the amyloidogenic fragments from the full-length gelsolin variants and demonstrated that heparin is capable of accelerating gelsolin amyloidogenesis. Herein, we identify the structural features of heparin that promote the 8 kDa disease-associated gelsolin fragments (residues 173-243) generated at the cell surface to form amyloid. In conjunction with electron microscopy analyses, our kinetic studies demonstrate that heparin efficiently accelerates the formation of gelsolin amyloid by enabling intermolecular beta-sheet formation. The use of heparin analogues reveals that sulfation is important in accelerating amyloidogenesis and that the extent of acceleration is proportional to the molecular weight of heparin. In addition, heparin accelerated aggregation at both early and late stages of amyloidogenesis. Dynamic light scattering coupled to size exclusion chromatography showed that heparin promotes the formation of soluble aggregates. Collectively, these data reveal that heparin templates fibril formation and affords solubility to the aggregating peptides through its sulfated structure. By extension, the biochemical results herein suggest that tissue-selective deposition characteristic of the gelsolin amyloidoses is likely influenced by the extracellular localization of distinct glycosaminoglycans. PMID- 16475812 TI - Membrane binding of twin arginine preproteins as an early step in translocation. AB - The twin arginine transport (Tat) system translocates folded proteins across the bacterial inner membrane. Transport substrates are recognized by means of evolutionarily well-conserved N-terminal signal peptides. The precise role of signal peptides in the actual transport process is not yet fully understood. Potentially, much insight into the molecular details of the transport process could be gained from step-by-step in vitro experiments under controlled conditions. Here, we employ purified preproteins to study their interaction with the phospholipid membrane by using surface plasmon resonance spectroscopy. It turns out that preproteins interact tightly with a model membrane consisting of only phospholipids. This interaction, which is stabilized by both electrostatic and hydrophobic contributions, appears to constitute an early step in protein translocation by the Tat system. PMID- 16475813 TI - Structural model of the BCL-w-BID peptide complex and its interactions with phospholipid micelles. AB - A peptide corresponding to the BH3 region of the proapoptotic protein, BID, could be bound in the cleft of the antiapoptotic protein, BCL-w. This binding induced major conformational rearrangements in both the peptide and protein components of the complex and led to the displacement and unfolding of the BCL-w C-terminal alpha-helix. The structure of BCL-w with a bound BID-BH3 peptide was determined using NMR spectroscopy and molecular docking. These studies confirmed that a region of 16 residues of the BID-BH3 peptide is responsible for its strong binding to BCL-w and BCL-x(L). The interactions of BCL-w and the BID-BH3 peptide complex with dodecylphosphocholine micelles were characterized and showed that the conformational change of BCL-w upon lipid binding occurred at the same time as the release and unfolding of the BH3 peptide. PMID- 16475815 TI - Double mutation at the subunit interface of glutathione transferase rGSTM1-1 results in a stable, folded monomer. AB - Canonical glutathione (GSH) transferases are dimeric proteins with subunits composed of an N-terminal GSH binding region (domain 1) and a C-terminal helical region (domain 2). The stabilities of several GSH transferase dimers are dependent upon two groups of interactions between domains 1 and 2 of opposing subunits: a hydrophobic ball-and-socket motif and a buried charge cluster motif. In rGSTM1-1, these motifs involve residues F56 and R81, respectively. The structural basis for the effects of mutating F56 to different residues on dimer stability and function has been reported (Codreanu et al. (2005) Biochemistry 44, 10605-10612). Here, we show that the simultaneous disruption of both motifs in the F56S/R81A mutant causes complete dissociation of the dimer to a monomeric protein on the basis of gel filtration chromatography and multiple-angle laser light scattering. The fluorescence and far-UV CD properties of the double mutant as well as the kinetics of amide H/D exchange along the polypeptide backbone suggest that the monomer has a globular structure that is similar to a single subunit in the native protein. However, the mutant monomer has severely impaired catalytic activity, suggesting that the dimer interface is vital for efficient catalysis. Backbone amide H/D exchange kinetics in the F56S and F56S/R81A mutants indicate that a reorganization of the loop structure between helix alpha2 and strand beta3 near the active site is responsible for the decreased catalytic activity of the monomer. In addition, the junction between the alpha4 and alpha5 helices in F56S/R81R shows decreased H/D exchange, indicating another structural change that may affect catalysis. Although the native subunit interface is important for dimer stability, urea-induced unfolding of the F56S/R81A mutant suggests that the interface is not essential for the thermodynamic stability of individual subunits. The H/D exchange data reveal a possible molecular basis for the folding cooperativity observed between domains 1 and 2. PMID- 16475814 TI - Identification of an ordered compact structure within the recombinant bovine fibrinogen alphaC-domain fragment by NMR. AB - The NMR solution structure of the bovine fibrinogen alphaC-domain fragment, including residues Aalpha374-538, reveals a type-I' beta-hairpin, restricted at the base by a C423-C453 disulfide linkage and a short turn preceding C423. Although both faces of the hairpin are formed mainly by hydrophilic residues, one of them is uncharged while the other has a characteristic pattern of charged residues which are highly conserved among vertebrate species. Chemical shift indexing and relaxation data indicate the presence of a collapsed hydrophobic region next to the hairpin that includes approximately 30 residues with slower concerted motion and higher content of nonpolar residues and, according to a previous study (Tsurupa, G., Tsonev, L., and Medved, L. (2002) Biochemistry 41, 6449-6459), may cooperate with the hairpin to form a compact cooperative unit (domain). Structure and relaxation data show that the region between C423 and C453 is populated by both random coil and beta-structure, suggesting that the cooperative structure in the isolated alphaC-domain is intrinsically unstable. This observation is in agreement with a very low energy of stabilization of the Aalpha374-538 fragment determined in unfolding experiments. The low stability of the alphaC-domain suggests a possible explanation for the previously observed intra- and intermolecular interactions of these domains in fibrinogen and fibrin. PMID- 16475816 TI - Sulfate-induced effects in the on-pathway intermediate of the bacterial immunity protein Im7. AB - Intermediates have now been identified in the folding of a number of small, single-domain proteins. Here we describe experiments to determine the effect of Na(2)SO(4) on the properties of the on-pathway intermediate formed early during the folding of the four-helical protein, Im7. This intermediate, studied previously in 0.4 M Na(2)SO(4), contains three of the four native helices and is fascinating in that several residues in helices I, II, and IV make non-native interactions that stabilize this state. Whether these contacts form as a consequence of the presence of Na(2)SO(4), however, remained unresolved. Using kinetic analysis of the effect of Na(2)SO(4) on the unfolding and refolding kinetics of Im7*, combined with detailed analysis of the resulting chevron plots, we show that decreasing the concentration of Na(2)SO(4) from 0.4 to 0 M destabilizes the intermediate and rate-limiting transition (TS2) states by 7 and 10 kJ mol(-)(1), respectively, and has little effect on the relative compactness of these states compared with that of the unfolded ensemble (beta(I) approximately 0.8, beta(TS2) approximately 0.9 in 0 to 0.4 M Na(2)SO(4)). Analysis of 10 variants of the protein in 0.2 M Na(2)SO(4) using Phi-values showed that the structural properties of the intermediate and TS2 are not altered significantly by the concentration of the kosmotrope. The data demonstrate that the rapid formation of a compact intermediate stabilized by non-native interactions during Im7* folding is not induced by high concentrations of the stabilizing salt, but is a generic feature of the folding of this protein. PMID- 16475817 TI - Global hairpin folding of tau in solution. AB - The microtubule-associated protein tau stabilizes microtubules in its physiological role, whereas it forms insoluble aggregates (paired helical filaments) in Alzheimer's disease. Soluble tau is considered a natively unfolded protein whose residual folding and intramolecular interactions are largely undetermined. In this study, we have applied fluorescence resonance energy transfer (FRET) and electron paramagnetic resonance (EPR) to examine the proximity and flexibility of tau domains and the global folding. FRET pairs spanning the tau molecule were created by inserting tryptophans (donor) and cysteines (labeled with IAEDANS as an acceptor) by site-directed mutagenesis. The observed FRET distances were significantly different from those expected for a random coil. Notably, the C-terminal end of tau folds over into the vicinity of the microtubule-binding repeat domain, the N-terminus remains outside the FRET distance of the repeat domain, yet both ends of the molecule approach one another. The interactions between the domains were obliterated by denaturation in GdnHCl. Paramagnetic spin-labels attached in various domains of tau were analyzed by EPR and exhibited a high mobility throughout. The data indicate that tau retains some global folding even in its "natively unfolded" state, combined with the high flexibility of the chain. PMID- 16475818 TI - Distinct targeting and recycling properties of two isoforms of the iron transporter DMT1 (NRAMP2, Slc11A2). AB - The metal transporter DMT1 (Slc11a2) plays a vital role in iron metabolism. Alternative splicing of the 3' exon generates two DMT1 isoforms with different C terminal protein sequences and a 3' untranslated region harboring (isoform I, +IRE) or not (isoform II, -IRE), an iron-responsive element. Isoform I is expressed at the plasma membrane of certain epithelial cells including the duodenum brush border, where it is essential for the absorption of nutritional iron. Isoform II is expressed in many cells and is essential for the acquisiton of transferrin iron from acidified endosomes. The targeting and trafficking properties of DMT1 isoforms I and II were studied in transfected LLC-PK(1) kidney cells, with respect to isoform-specific differences in function, subcellular localization, endocytosis kinetics, and fate upon internalization. Isoform I showed higher surface expression and was internalized from the plasma membrane with slower kinetics than that of isoform II. As opposed to isoform II, which is efficiently sorted to recycling endosomes upon internalization, isoform I was not efficiently recycled and was targeted to lysosomes. Thus, alternative splicing of DMT1 critically regulates the subcellular localization and site of Fe(2+) transport. PMID- 16475819 TI - Transmembrane helices 3 and 4 are involved in substrate recognition by the Na+/dicarboxylate cotransporter, NaDC1. AB - The Na(+)/dicarboxylate cotransporters (NaDC1) from mouse (m) and rabbit (rb) differ in their ability to handle glutarate. Substrate-dependent inward currents, measured using two-electrode voltage clamp, were similar for glutarate and succinate in Xenopus oocytes expressing mNaDC1. In contrast, currents evoked by glutarate in rbNaDC1 were only about 5% of the succinate-dependent currents. To identify domains involved in glutarate transport, we constructed a series of chimeric transporters between mouse and rabbit NaDC1. Although residues found in multiple transmembrane helices (TM) participate in glutarate transport, the most important contribution is made by TM 3 and 4 and the associated loops. The R(M3 4) chimera, consisting of rbNaDC1 with substitution of TM 3-4 from mNaDC1, had a decreased K(0.5)(glutarate) of 4 mM compared with 15 mM in wild-type rbNaDC1 without any effect on K(0.5)(succinate). The chimeras were also characterized using dual-label competitive uptakes with (14)C-glutarate and (3)H-succinate to calculate the transport specificity ratio (TSR), a measure of relative catalytic efficiency with the two substrates. The TSR analysis provides evidence for functional coupling in the transition state between TM 3 and 4. We conclude that TM 3 and 4 contain amino acid residues that are important determinants of substrate specificity and catalytic efficiency in NaDC1. PMID- 16475820 TI - A systematic study of the effect of physiological factors on beta2-microglobulin amyloid formation at neutral pH. AB - Beta(2)-microglobulin (beta(2)m) forms amyloid fibrils that deposit in the musculo-skeletal system in patients undergoing long-term hemodialysis. How beta(2)m self-assembles in vivo is not understood, since the monomeric wild-type protein is incapable of forming fibrils in isolation in vitro at neutral pH, while elongation of fibril-seeds made from recombinant protein has only been achieved at low pH or at neutral pH in the presence of detergents or cosolvents. Here we describe a systematic study of the effect of 11 physiologically relevant factors on beta(2)m fibrillogenesis at pH 7.0 without denaturants. By comparing the results obtained for the wild-type protein with those of two variants (DeltaN6 and V37A), the role of protein stability in fibrillogenesis is explored. We show that DeltaN6 forms low yields of amyloid-like fibrils at pH 7.0 in the absence of seeds, suggesting that this species could initiate fibrillogenesis in vivo. By contrast, high yields of amyloid-like fibrils are observed for all proteins when assembly is seeded with fibril-seeds formed from recombinant protein at pH 2.5 stabilized by the addition of heparin, serum amyloid P component (SAP), apolipoprotein E (apoE), uremic serum, or synovial fluid. The results suggest that the conditions within the synovium facilitate fibrillogenesis of beta(2)m and show that different physiological factors may act synergistically to promote fibril formation. By comparing the behavior of wild type beta(2)m with that of DeltaN6 and V37A, we show that the physiologically relevant factors enhance fibrillogenesis by stabilizing fibril-seeds, thereby allowing fibril extension by rare assembly competent species formed by local unfolding of native monomers. PMID- 16475821 TI - Phenylalanine 90 and 93 are localized within the phenol binding site of human UDP glucuronosyltransferase 1A10 as determined by photoaffinity labeling, mass spectrometry, and site-directed mutagenesis. AB - 4-Azido-2-hydroxybenzoic acid (4-AzHBA), a novel photoactive benzoic acid derivative, has been synthesized and used as a photoprobe to identify the phenol binding site of UDP-glucuronosyltransferases (UGTs). Analysis of recombinant His tag UGTs from the 1A family for their ability to glucuronidate p-nitrophenol (pNP) and 4-methylumbelliferone (4-MU) revealed that UGT1A10 shows high activity toward phenols and phenol derivatives. Purified UGT1A10 was photolabeled with 4 AzHBA, digested with trypsin, and analyzed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF)-mass spectrometry. A single modified peak corresponding to amino acid residues 89-98 (EFMVFHAQWK) of UGT1A10 was identified. The attachment site of the 4-AzHBA probe was localized to the quadruplet Phe(90)-Met(91)-Val(92)-Phe(93) using ESI LC-MS/MS. Sequence alignment revealed that the Phe(90) and Phe(93) are conserved in UGT1A7-10. Site-directed mutagenesis of these two amino acids was then followed by kinetic analysis of the mutants with two phenolic substrates, pNP and 4-MU, containing one and two planar rings, respectively. Using the combination of photoaffinity labeling, enzymatic digestion, MALDI-TOF and LC-MS mass spectrometry, and site-directed mutagenesis, we have determined for the first time that Phe(90) and Phe(93) are directly involved in the catalytic activity of UGT1A10 toward 4-MU and pNP. PMID- 16475823 TI - Biochemical indication for myristoylation-dependent conformational changes in HIV 1 Nef. AB - The accessory HIV-1 Nef protein is essential for viral replication, high virus load, and progression to AIDS. These functions are mediated by the alteration of signaling and trafficking pathways and require the membrane association of Nef by its N-terminal myristoylation. However, a large portion of Nef is also found in the cytosol, in line with the observation that myristoylation is only a weak lipidation anchor for membrane attachment. We performed biochemical studies to analyze the implications of myristoylation on the conformation of Nef in aqueous solution. To establish an in vivo myristoylation assay, we first optimized the codon usage of Nef for Escherichia coli expression, which resulted in a 15-fold higher protein yield. Myristoylation was achieved by coexpression with the N myristoyltransferase and confirmed by mass spectrometry. The myristoylated protein was soluble, and proton NMR spectra confirmed proper folding. Size exclusion chromatography revealed that myristoylated Nef appeared of smaller size than the unmodified form but not as small as an N-terminally truncated from of Nef that omits the anchor domain. Western blot stainings and limited proteolysis of both forms showed different recognition profiles and degradation pattern. Analytical ultracentrifugation revealed that myristoylated Nef prevails in a monomeric state while the unmodified form exists in an oligomeric equilibrium of monomer, dimer, and trimer associations. Finally, fluorescence correlation spectroscopy using multiphoton excitation revealed a shorter diffusion time for the lipidated protein compared to the unmodified form. Taken together, our data indicated myristoylation-dependent conformational changes in Nef, suggesting a rather compact and monomeric form for the lipidated protein in solution. PMID- 16475822 TI - Thermodynamics of protein-protein interactions of cMyc, Max, and Mad: effect of polyions on protein dimerization. AB - The Myc-Max-Mad network of proteins activates or represses gene transcription depending on whether the dimerization partner of Max is c-Myc or Mad. To elucidate the physical properties of these protein-protein interactions, fluorescence anisotropy of TRITC-labeled Max was used. The binding affinities and thermodynamics of dimerization of the Max-Max homodimer and c-Myc-Max and Mad-Max heterodimers were determined. Our results indicate that c-Myc and Max form the most stable heterodimer. Previous work [Kohler, J. J., Metallo, S. J., Schneider, T. L., and Schepartz, A. (1999) Proc. Natl. Acad. Sci. U.S.A. 96, 11735-9] has shown that instead of dimerizing first and then binding to DNA, these proteins use a monomer pathway in which a monomer binds to DNA followed by dimerization on the surface of the DNA. The DNA E-box affects the dimerization, but nonspecific effects may also play a role. The influence of polyions, poly-L-lysine and poly-L glutamic acid, were investigated to determine the effects of charged polymers other than DNA on homodimerization and heterodimerization. While the positively charged poly-L-lysine, PLL, did not show any significant effect, negatively charged poly-L-glutamic acid, PLG, stabilized both heterodimers and homodimers by 2-3 kJ/mol. These data suggest that in the cell nucleus the presence of negatively charged DNA or RNA could nonspecifically aid in association of these proteins. Calculations of DeltaH degrees and DeltaS degrees from the temperature dependence of K(d) indicated that although the thermodynamic parameters for the dimer are different, the reactions for all three dimers are driven by negative (favorable) enthalpic and negative (unfavorable) entropic contributions. In the presence of PLG, entropy became more negative with the effect being largest for c Myc-Max heterodimers. This suggests that van der Waals and H-bonding interactions are predominant in dimerization of these proteins. PMID- 16475824 TI - Glycoform quantification of chondroitin/dermatan sulfate using a liquid chromatography-tandem mass spectrometry platform. AB - Chondroitin sulfate (CS) is a glycosaminoglycan consisting of repeating uronic acid, N-acetylgalactosamine disaccharide units {[HexAbeta/alpha(1-3)GalNAcbeta(1 4)](n)()}. CS chains are polydisperse with respect to chain length, sulfate content, and glucuronic acid epimerization content, resulting in a distribution of glycoforms for a chain bound to any given serine residue. Usually, CS glycoforms exist, differing in sulfation position and uronic acid epimerization. This work introduces a novel LC-MS/MS platform for the quantification of mixtures of CS oligosaccharides. The CS polysaccharides were partially depolymerized and labeled with either the light (d(0)) or heavy (d(4)) form of 2-anthranilic acid (2-AA). Excess reagent was removed, and mixtures of the CS standard (d(0)) and unknown (d(4)) were made. The CS mixture was subjected to size exclusion chromatography (SEC) with on-line electrospray ionization mass spectrometric detection in the negative ion mode. Tandem mass spectra were acquired, and quantification of unknown samples within the mixture was made using relative ion abundances of specific diagnostic ions. The high accuracy and precision of the glycomics platform were demonstrated using glycoform mixtures made from standard CS preparations. The CS glycoform analysis method was then applied to cartilage extract, versican, and several dermatan sulfate preparations. This work presents the first application of a glycomics platform for the quantification of CS oligosaccharide mixtures for obtaining specific information about the positions of GalNAc sulfation and uronic acid epimerization. PMID- 16475825 TI - Mechanistic insight provided by glutaredoxin within a fusion to redox-sensitive yellow fluorescent protein. AB - Redox-sensitive yellow fluorescent protein (rxYFP) contains a dithiol disulfide pair that is thermodynamically suitable for monitoring intracellular glutathione redox potential. Glutaredoxin 1 (Grx1p) from yeast is known to catalyze the redox equilibrium between rxYFP and glutathione, and here, we have generated a fusion of the two proteins, rxYFP-Grx1p. In comparison to isolated subunits, intramolecular transfer of reducing equivalents made the fusion protein kinetically superior in reactions with glutathione. The rate of GSSG oxidation was thus improved by a factor of 3300. The reaction with GSSG most likely takes place entirely through a glutathionylated intermediate and not through transfer of an intramolecular disulfide bond. However, during oxidation by H(2)O(2), hydroxyethyl disulfide, or cystine, the glutaredoxin domain reacted first, followed by a rate-limiting (0.13 min(-)(1)) transfer of a disulfide bond to the other domain. Thus, reactivity toward other oxidants remains low, giving almost absolute glutathione specificity. We have further studied CPYC --> CPYS variants in the active site of Grx1p and found that the single Cys variant had elevated oxidoreductase activity separately and in the fusion. This could not be ascribed to the lack of an unproductive side reaction to glutaredoxin disulfide. Instead, slower alkylation kinetics with iodoacetamide indicates a better leaving-group capability of the remaining cysteine residue, which can explain the increased activity. PMID- 16475827 TI - Assembly and disassembly kinetics of anthrax toxin complexes. AB - Proteolytic activation of the protective antigen (PA) component of anthrax toxin allows it to self-associate into a ring-shaped homoheptamer, [PA(63)](7), which can bind the enzymatic components lethal factor (LF) and edema factor (EF). [PA(63)](7) is a pore-precursor (prepore), and under the low-pH conditions of the endosome, it forms a transmembrane pore that allows LF and EF to enter the cytosol. PA was labeled with donor and acceptor fluorescent dyes, and Forster resonance energy transfer was used to measure the assembly and disassembly kinetics of the prepore complex in solution. The dissociation rate constant for [PA(63)](7) was 1 x 10(-)(6) s(-)(1) (t(1/2) approximately 7 days). In contrast, a ternary complex containing the PA-binding domain of LF (LF(N)) bound to a PA(63) dimer composed of two nonoligomerizing mutants dissociated rapidly (t(1/2) approximately 1 min). Thus, the substantial decrease in the rate of disassembly of [PA(63)](7) relative to the ternary complex is due to the cooperative interactions among neighboring subunits in the heptameric ring. Low concentrations of LF(N) promoted assembly of the prepore from proteolytically activated PA, whereas high concentrations inhibited assembly of both the prepore and the ternary complex. A self-assembly scheme of anthrax toxin complexes is proposed. PMID- 16475829 TI - Differential phosphorylation of the rhodopsin cytoplasmic tail mediates the binding of arrestin and its splice variant, p44. AB - Deactivation of the vertebrate photopigment rhodopsin is achieved through a two step process. Rhodopsin is first phosphorylated by rhodopsin kinase and subsequently deactivated by the binding of the regulatory protein arrestin or its splice variant, p44. Although much is known about the overall differences between arrestin and p44 binding to different rhodopsin species (photolyzed versus unphotolyzed and/or phosphorylated versus unphosphorylated), the exact role of p44 during phototransduction remains to be fully elucidated. Our current study addresses this question by identifying structural differences between arrestin and p44 and characterizing the interaction between the negatively charged rhodopsin tail and either p44 or arrestin. Our results demonstrate that arrestin and p44 bind differently to different phosphorylated rhodopsin species and that this may be due to a structural difference between p44's and arrestin's basal states. This difference offers a potential regulatory mechanism that could regulate p44 and arrestin binding and, as a result, regulate the kinetics of the rod's light response. PMID- 16475826 TI - Reduction and oxidation of the active site iron in tyrosine hydroxylase: kinetics and specificity. AB - Tyrosine hydroxylase (TyrH) is a pterin-dependent enzyme that catalyzes the hydroxylation of tyrosine to form dihydroxyphenylalanine. The oxidation state of the active site iron atom plays a central role in the regulation of the enzyme. The kinetics of reduction of ferric TyrH by several reductants were determined by anaerobic stopped-flow spectroscopy. Anaerobic rapid freeze-quench EPR confirmed that the change in the near-UV absorbance of TyrH upon adding reductant corresponded to iron reduction. Tetrahydrobiopterin reduces wild-type TyrH following a simple second-order mechanism with a rate constant of 2.8 +/- 0.1 mM( )(1) s(-)(1). 6-Methyltetrahydropterin reduces the ferric enzyme with a second order rate constant of 6.1 +/- 0.1 mM(-)(1) s(-)(1) and exhibits saturation kinetics. No EPR signal for a radical intermediate was detected. Ascorbate, glutathione, and 1,4-benzoquinone all reduce ferric TyrH, but much more slowly than tetrahydrobiopterin, suggesting that the pterin is a physiological reductant. E332A TyrH, which has an elevated K(m) for tetrahydropterin in the catalytic reaction, is reduced by tetrahydropterins with the same kinetic parameters as those of the wild-type enzyme, suggesting that BH(4) does not bind in the catalytic conformation during the reduction. Oxidation of ferrous TyrH by molecular oxygen can be described as a single-step second-order reaction, with a rate constant of 210 mM(-)(1) s(-)(1). S40E TyrH, which mimics the phosphorylated state of the enzyme, has oxidation and reduction kinetics similar to those of the wild-type enzyme, suggesting that phosphorylation does not directly regulate the interconversion of the ferric and ferrous forms. PMID- 16475828 TI - Glycerol monolaurate inhibits the effects of Gram-positive select agents on eukaryotic cells. AB - Many exotoxins of Gram-positive bacteria, such as superantigens [staphylococcal enterotoxins, toxic shock syndrome toxin-1 (TSST-1), and streptococcal pyrogenic exotoxins] and anthrax toxin are bioterrorism agents that cause diseases by immunostimulation or cytotoxicity. Glycerol monolaurate (GML), a fatty acid monoester found naturally in humans, has been reported to prevent synthesis of Gram-positive bacterial exotoxins. This study explored the ability of GML to inhibit the effects of exotoxins on mammalian cells and prevent rabbit lethality from TSS. GML (>or=10 microg/mL) inhibited superantigen (5 microg/mL) immunoproliferation, as determined by inhibition of (3)H-thymidine incorporation into DNA of human peripheral blood mononuclear cells (1 x 10(6) cells/mL) as well as phospholipase Cgamma1, suggesting inhibition of signal transduction. The compound (20 microg/mL) prevented superantigen (100 microg/mL) induced cytokine secretion by human vaginal epithelial cells (HVECs) as measured by ELISA. GML (250 microg) inhibited rabbit lethality as a result of TSST-1 administered vaginally. GML (10 microg/mL) inhibited HVEC and macrophage cytotoxicity by anthrax toxin, prevented erythrocyte lysis by purified hemolysins (staphylococcal alpha and beta) and culture fluids containing streptococcal and Bacillus anthracis hemolysins, and was nontoxic to mammalian cells (up to 100 microg/mL) and rabbits (250 microg). GML stabilized mammalian cell membranes, because erythrocyte lysis was reduced in the presence of hypotonic aqueous solutions (0 0.05 M saline) or staphylococcal alpha- and beta-hemolysins when erythrocytes were pretreated with GML. GML may be useful in the management of Gram-positive exotoxin illnesses; its action appears to be membrane stabilization with inhibition of signal transduction. PMID- 16475831 TI - A novel carbonic anhydrase II binding site regulates NHE1 activity. AB - Carbonic anhydrase II (CAII) binds to and regulates transport by the NHE1 isoform of the mammalian Na(+)/H(+) exchanger. We localized and characterized the CAII binding region on the C-terminal tail of the Na(+)/H(+) exchanger. CAII did not bind to acidic sequences in NHE1 that were similar to the CAII binding site of bicarbonate transporters. Instead, by expressing a variety of fusion proteins of the C-terminal region of the Na(+)/H(+) exchanger, we demonstrated that CAII binds to the penultimate group of 13 amino acids of the cytoplasmic tail. Within this region, site-specific mutagenesis demonstrated that amino acids S796 and D797 form part of a novel CAII binding site. Phosphorylation of the C-terminal 26 amino acids by heart cell extracts did not alter CAII binding to this region, but phosphorylation greatly increased CAII binding to a protein containing the C terminal 182 amino acids of NHE1. This suggested that an upstream region of the cytoplasmic tail acts as an inhibitor of CAII binding to the penultimate group of 13 amino acids. The results demonstrate that a novel phosphorylation-regulated CAII binding site exists in distal amino acids of the NHE1 tail. PMID- 16475832 TI - Characterization of Cdr1p, a major multidrug efflux protein of Candida albicans: purified protein is amenable to intrinsic fluorescence analysis. AB - Candida drug resistance protein 1 (Cdr1p), an ATP-dependent drug efflux pump, confers multidrug resistance in immunocompromised and debilitated patients. A member of the ATP-binding cassette (ABC) superfamily of membrane transporters, Cdr1p contains two nucleotide binding/utilization sites (NBDs) and two transmembrane domains (TMDs). We had earlier characterized Cdr1p by its overexpression as a GFP-tagged fusion protein that elicits oligomycin-sensitive ATPase activity and is linked to drug extrusion. However, it is essential to have highly purified Cdr1p to understand the detailed molecular basis of structure and functions of this protein. In this study, we have developed a two-step purification protocol using stably overexpressed His-tagged Cdr1p in Saccharomyces cerevisiae. Purified Cdr1p exhibited divalent cation-dependent ATPase activity [approximately 1.2 micromol (mg of protein)(-)(1) min(-)(1)] with an apparent K(M) in the range of 1.8 to 2.1 mM and V(max) between 1.0 and 1.4 micromol (mg of protein)(-)(1) min(-)(1). Unlike its close homologue human P gp/MDR1, purified Cdr1p only moderately displayed drug stimulated ATPase activity. By exploiting intrinsic fluorescence intensity of purified Cdr1p, which contains 24 tryptophan residues, we could monitor defined conformational changes upon substrate drug and ATP binding. It is observed that ATP binding to Cdr1p (K(d) = approximately 1.7 mM) is not a prerequisite for drug binding, and both the mechanisms of drug as well as ATP binding, which induce specific conformational changes, occur independent of each other. Our study for the first time provides a catalytically active purified ABC transporter from a fungal pathogen, which is amenable to fluorescence measurements and thus would be useful in understanding the molecular basis of antifungal transport. PMID- 16475830 TI - Inhibition of apolipoprotein AI gene expression by tumor necrosis factor alpha: roles for MEK/ERK and JNK signaling. AB - Plasma high-density lipoprotein and apolipoprotein AI (apoAI) levels are suppressed by tumor necrosis factor alpha. To determine the molecular mechanisms responsible for the effect of TNF alpha on the apoAI promoter activity, HepG2 cells were exposed to both genetic and pharmacological modulators of TNF alpha mediated signaling in the presence or absence of TNF alpha. Exogenous ERK1 and ERK2 expression suppressed basal apoAI promoter activity; however, only ERK2 enhanced the ability of TNF alpha to suppress apoAI promoter activity. Exogenous expression of all three MEK isoforms (MEK1, MEK2A, and MEK2E) suppressed basal apoAI promoter activity and further aggravated TNF alpha-related apoAI promoter activity inhibition. Treatment with SB202190 (p38 MAP kinase inhibitor) alone significantly increased apoAI promoter activity; however, in the presence of TNF alpha, apoAI promoter activity was suppressed to an extent similar to that in cells not treated with SB202190. ApoAI promoter activity increased in cells treated with the specific JNK inhibitor SP600125, but unlike SB202190 treatment, the level of TNF alpha-related apoAI promoter inhibition was reduced by 50%. Similarly, the level of TNF alpha-related apoAI promoter inhibition was reduced in cells transfected with JNK1 siRNA. Finally, treatment of cells with the NF kappaB inhibitors BAY and SN-50 or overexpression of NF-kappaB subunits p50 and p65 had no effect on the ability of TNF alpha to repress apoAI promoter activity. These results suggest that TNF alpha suppresses apoAI promoter activity through both the MEK/ERK and JNK pathways but is not mediated by either p38 MAP kinase activity or NF-kappaB activation. PMID- 16475833 TI - Charge separation kinetics in intact photosystem II core particles is trap limited. A picosecond fluorescence study. AB - The fluorescence kinetics in intact photosystem II core particles from the cyanobacterium Thermosynechococcus elongatus have been measured with picosecond resolution at room temperature in open reaction centers. At least two new lifetime components of approximately 2 and 9 ps have been resolved in the kinetics by global analysis in addition to several known longer-lived components (from 42 ps to approximately 2 ns). Kinetic compartment modeling yields a kinetic description in full agreement with the one found recently by femtosecond transient absorption spectroscopy [Holzwarth et al. (2005) submitted to Proc. Natl. Acad. Sci. U.S.A.]. We have for the first time resolved directly the fluorescence spectrum and the kinetics of the equilibrated excited reaction center in intact photosystem II and have found two early radical pairs before the electron is transferred to the quinone Q(A). The apparent lifetime for primary charge separation is 7 ps, that is, by a factor of 8-12 faster than assumed on the basis of earlier analyses. The main component of excited-state decay is 42 ps. The effective primary charge separation rate constant is 170 ns(-)(1), and the secondary electron-transfer rate constant is 112 ns(-)(1). Both electron transfer steps are reversible. Electron transfer from pheophytin to Q(A) occurs with an apparent overall lifetime of 350 ps. The energy equilibration between the CP43/CP47 antenna and the reaction center occurs with a main apparent lifetime of approximately 1.5 ps and a minor 10 ps lifetime component. Analysis of the overall trapping kinetics based on the theory of energy migration and trapping on lattices shows that the charge separation kinetics in photosystem II is extremely trap-limited and not diffusion-to-the-trap-limited as claimed in several recent papers. These findings support the validity of the assumptions made in deriving the earlier exciton radical pair equilibrium model [Schatz, G. H., Brock, H., and Holzwarth, A. R. (1988) Biophys. J. 54, 397-405]. PMID- 16475834 TI - Mechanistic studies of ubiquitin C-terminal hydrolase L1. AB - Ubiquitin C-terminal hydrolases (UCHs) cleave Ub-X bonds (Ub is ubiquitin and X an alcohol, an amine, or a protein) through a thioester intermediate that is produced by nucleophilic attack of the Cys residue of a Cys-SH/His-Im catalytic diad. We are studying the mechanism of UCH-L1, a UCH that is implicated in Parkinson's disease, and now wish to report our initial findings. (i) Pre-steady state kinetic studies for UCH-L1-catalyzed hydrolysis of Ub-AMC (AMC, 7-amido-4 methylcoumarin) indicate that k(cat) is rate-limited by acyl-enzyme formation. Thus, K(m) = K(s), the dissociation constant for the Michaelis complex, and k(cat) = k(2), the rate constant for acyl-enzyme formation. (ii) For K(assoc) (=K(s)(-)(1)), DeltaC(p) = -0.8 kcal mol(-)(1) deg(-)(1) and is consistent with coupling between substrate association and a conformational change of the enzyme. For k(2), DeltaS(++) = 0 and suggests that in the E-S, substrate and active site residues are precisely aligned for reaction. (iii) Solvent isotope effects are (D)K(assoc) = 0.5 and (D)k(2) = 0.9, suggesting that the substrate binds to a form of free enzyme in which the active site Cys exists as the thiol. In the resultant Michaelis complex, the diad has tautomerized to ion pair Cys-S(-)/His ImH(+). Subsequent attack of thiolate produces the acyl-enzyme species. In contrast, isotope effects for association of UCH-L1 with transition-state analogue ubiquitin aldehyde suggest that an alternative mechanistic pathway can sometimes be available to UCH-L1 involving general base-catalyzed attack of Cys SH by His-Im. PMID- 16475835 TI - Optimum activity of the phosphofructokinase from Ascaris suum requires more than one metal ion. AB - Phosphofructokinase (PFK) catalyzes the phosphorylation of fructose 6-phosphate (F6P) to give fructose 1,6-bisphosphate (FBP) using MgATP as the phosphoryl donor. As the concentration of Mg(2+) increases above the concentration needed to generate the MgATP chelate complex, a 15-fold increase in the initial rate was observed at low MgATP. The effect of Mg(2+) is limited to V/K(MgATP), and initial rate studies indicate an equilibrium-ordered addition of Mg(2+) before MgATP. Isotope partitioning of the dPFK:MgATP complex indicates a random addition of MgATP and F6P at low Mg(2+), with the rate of release of MgATP from the central E:MgATP:F6P complex 4-fold faster than the net rate constant for catalysis. This can be contrasted with the ordered addition of MgATP prior to F6P at high Mg(2+). The addition of fructose 2,6-bisphosphate (F26P(2)) has no effect on the mechanism at low Mg(2+), with the exception of a 4-fold increase in the affinity of the enzyme for F6P. At high Mg(2+), F26P(2) causes the kinetic mechanism to become random with respect to MgATP and F6P and with MgATP released from the central complex half as fast as the net rate constant for catalysis. The latter is in agreement with previous studies [Gibson, G. E., Harris, B. G., and Cook, P. F. (1996) Biochemistry 35, 5451-5457]. The overall effect of Mg(2+) is a decrease in the rate of release of MgATP from the E:MgATP:F6P complex, independent of the concentration of F26P(2). PMID- 16475837 TI - The future of computing--new architectures and new technologies. AB - All modern computers are designed using the 'von Neumann' architecture and built using silicon transistor technology. Both architecture and technology have been remarkably successful. Yet there are a range of problems for which this conventional architecture is not particularly well adapted, and new architectures are being proposed to solve these problems, in particular based on insight from nature. Transistor technology has enjoyed 50 years of continuing progress. However, the laws of physics dictate that within a relatively short time period this progress will come to an end. New technologies, based on molecular and biological sciences as well as quantum physics, are vying to replace silicon, or at least coexist with it and extend its capability. The paper describes these novel architectures and technologies, places them in the context of the kinds of problems they might help to solve, and predicts their possible manner and time of adoption. Finally it describes some key questions and research problems associated with their use. PMID- 16475836 TI - Allosteric aptamers controlling a signal amplification cascade allow visual detection of molecules at picomolar concentrations. AB - A broadly applicable homogeneous detection system has been developed. It utilizes components of the blood coagulation cascade in the presence of polystyrene microspheres (MS) as a signal amplifier. Russell's viper venom factor X activator (RVV-X) triggers the cascade, which results in an eye-visible phase transition (precipitation) of MS bound to clotted fibrin. An allosteric RNA aptamer, RNA132, with affinity for RVV-X and human vascular endothelial growth factor (VEGF(165)) was created. RNA132 inhibits enzymatic activity of RVV-X. The effector molecule, VEGF(165), reverses the inhibitory activity of RNA132 on RVV-X and restores its enzymatic activity, thus, triggering the cascade and enabling the phase transition. As few as 5 fmol of VEGF(165) could be detected by the naked eye within an hour. Similar results were obtained for another allosteric aptamer modulated by a protein tyrosine phosphatase. The assay is instrumentation-free for both processing and readout and can be modified to detect molecules to which aptamers can be obtained. PMID- 16475838 TI - Analysis of cellular structure by light scattering measurements in a new cytometer design based on a liquid-core waveguide. AB - The results of applying a novel microfluidic optical cytometer to generate and observe the light scattered from biological cells over a wide range of angles are presented. This cytometer incorporates a waveguide that increases the intensity of the scattered light to the extent that an inexpensive digital camera can be used to detect the light over a large solid angle. This device was applied to yeast cells and latex beads and experimental data were compared with the results of a finite difference time-domain (FDTD) method of simulation. The simulated scattering patterns were calculated from reported values of optical parameters and are in good qualitative agreement with experiment. It is demonstrated that this system could be used to acquire information on the microstructure and potentially the nanostructure of cells. PMID- 16475839 TI - Kv channel S6 helix as a molecular switch: simulation studies. AB - Ion channels form pores of nanoscopic dimensions in biological membranes and play a key role in the physiology of cells. The majority of ion channels are gated, i.e. they contain a molecular switch that allows a transition between a closed (functionally 'off') and open (functionally 'on') state. Comparison of crystal structures of potassium channels suggest that the gating mechanism of voltage gated potassium (Kv) channels involves a key role for the pore-lining S6 helix. There is a conserved PVP sequence motif in the S6 helix. Molecular dynamics simulations are used here to explore the conformational dynamics of the S6 helix hinge in models of fragments of a Kv channel, namely an S5-P-S6 monomer and an (S5-P-S6)4 tetramer. The latter is a model of the complete pore-forming domain of a Kv channel. All models were simulated embedded in an octane slab (a simple membrane mimetic). The results of these simulations indicate that the PVP motif may form a molecular hinge, even when the S6 helix forms part of a more complex model. The conformational dynamics of S6 are modulated by the remainder of protein, but it remains flexible. These simulation results are compatible with a channel gating model in which S6 bends in the vicinity of the PVP motif in addition to the region around the conserved glycine (G466) that is N-terminal to the PVP motif. This model is supported by comparison of the Kv S6 models with the S6 helix of the bacterial KvAP channel crystal structure. Thus, K channel gating may depend on a complex nanoswitch with three rigid helical sections linked by two molecular hinges. PMID- 16475840 TI - Adaptive, integrated sensor processing to compensate for drift and uncertainty: a stochastic 'neural' approach. AB - An adaptive stochastic classifier based on a simple, novel neural architecture- the Continuous Restricted Boltzmann Machine (CRBM) is demonstrated. Together with sensors and signal conditioning circuits, the classifier is capable of measuring and classifying (with high accuracy) the H+ ion concentration, in the presence of both random noise and sensor drift. Training on-line, the stochastic classifier is able to overcome significant drift of real incomplete sensor data dynamically. As analogue hardware, this signal-level sensor fusion scheme is therefore suitable for real-time analysis in a miniaturised multisensor microsystem such as a Lab-in-a-Pill (LIAP). PMID- 16475841 TI - Scanning probe technology in metalloprotein and biomolecular electronics. AB - The interfacing of man-made electronic components with specifically-folded biomacromolecules lies central not only to the development of sensory interfaces and potential new molecular-scale devices, but also enables us to analyse processes of great biological importance in a refined and controllable manner. Recent advances in both available technology, most notably optical and scanning probes in nature, and our understanding of suitable methodologies, have led us to the point where the characteristics of single biological molecules can be interrogated with good levels of reproducibility. We review here the application of scanning probe microscopy to the analysis of and experimentation on biological redox systems. Within this paper the tunnel transport characteristics, as assayed by both scanning tunnelling microscopy (STM) and conducting probe atomic force microscopy (AFM), of single metalloproteins are discussed. In a specific case study the electron transfer characteristics of the blue copper metalloprotein, azurin, are reported. The modulation of these properties under the influence of calibratable compressional force has also been examined in some detail. Work such as this enables one to reproducibly establish the conductance, barrier height, environmental sensitivity and electromechanical properties of these molecules. PMID- 16475842 TI - Novel molecular device based on electrostatic interactions in organic polymers. AB - A number of researchers have reported attempts to design molecular level devices. One approach is to make use of electrostatic interactions in different parts of a polymeric molecule. This paper reports a means to achieve this by adding space charge to a molecule consisting of symmetric and asymmetric subgroups. Physically, space charge residing in a subgroup produces a dipolar charge layer thereby creating a potential trough in the polymer backbone. By lifting or lowering this potential minimum, it is possible to modify the terminal current. The effect of space charge on the potential profile in the polymer backbone was examined and the change correlated to data on carrier mobilities for OC1C10-PPV reported in the literature. Modulation of space charge in the subgroup allows the manipulation of current flow along the polymer backbone, forming the basis for the development of a molecular device. A first-order analysis suggested that such a device could have current-voltage (I-V) characteristics similar to those of a MOSFET at subthreshold, with an estimated transconductance approximately 1-2 pAV and a cutoff frequency approximately 10(15) Hz. PMID- 16475843 TI - Cell response to nano-islands produced by polymer demixing: a brief review. AB - This review looks at the present literature available regarding cell response to nano-islands produced by nanotopography. Polymer demixing is a chemical method of fabricating large areas of nanotopography quickly and cheaply, making it ideal for cell testing and thus allowing it to be one of the first well-researched methods in cell engineering. The review shows that cells respond strongly to the islands (cell types observed include endothelial cells, fibroblasts, osteoblasts, leucocytes and platelets). Such changes include differences in adhesion, growth, gene expression and morphology. PMID- 16475844 TI - Formation of focal adhesion-stress fibre complexes coordinated by adhesive and non-adhesive surface domains. AB - Cell motility consists of repeating cycles of protrusion of a leading edge in the direction of migration, attachment of the advancing membrane to the matrix, and pulling of the trailing edge forward. In this dynamic process there is a major role for the cytoskeleton, which drives the protrusive events via polymerisation of actin in the lamellipodium, followed by actomyosin contractility. To study the transition of the actin cytoskeleton from a 'protrusive' to 'retractive' form, we have monitored the formation of focal adhesions and stress fibres during cell migration on a micro-patterned surface. This surface consisted of parallel arrays of 2 microm-wide, fibronectin-coated gold stripes, separated by non-adhesive (poly(ethylene glycol)-coated) glass areas with variable width, ranging from 4-12 microm. Monitoring the spreading of motile cells indicated that cell spreading was equally effective along and across the adhesive stripes, as long as the non adhesive spaces between them did not exceed 6 microm. When the width of the PEG region was 8 microm or more, cells became highly polarised upon spreading, and failed to reach the neighboring adhesive stripes. It was also noted that as soon as the protruding lamella successfully crossed the PEG-coated area and reached an adhesive region, the organisation of actin in that area was transformed from a diffuse meshwork into a bundle, oriented perpendicularly to the stripes and anchored at its ends in focal adhesions. This transition depends on actomyosin based contractility and is apparently triggered by the adhesion to the rigid fibronectin surface. PMID- 16475845 TI - Nanoscale surface engineered living cells with extended substrate spectrum. AB - We report on cell surface engineering of living microorganisms by using Layer-by Layer (LbL) technology to extend the substrate spectrum. The yeast Arxula adeninivorans LS3 (Arxula) was employed as a model organism and biological template. By using LbL technology, Arxula cells were encapsulated by polyelectrolyte and enzyme layers. The biological activity of the Arxula was retained after the encapsulation process. The polymeric capsule surrounding the Arxula provides a stable interface for surface engineering of living cells. LbL of polyelectrolytes followed by an enzyme layer of lactate oxidase were assembled. The outer enzyme layer provides an additional biological function for Arxula to convert the unfavourable substrate lactate into the favourable substrate pyruvate, thus extending the substrate spectrum of the organism. Moreover, capsule stability and enzyme conjugate stability of the surface engineered Arxula were studied. PMID- 16475846 TI - Biology and physics at interfaces: from single molecules to cells. PMID- 16475847 TI - Osteoblast cell membrane hybrid bilayers for studying cell-cell interactions. AB - Osteoblast-like cells were grown on a surface that presents cell membrane components to the cells in culture. The culture surface was a bimolecular layer formed by the interaction of osteoblast plasma membrane vesicles with an alkanethiol monolayer. The potential of these osteoblast-membrane hybrid bilayers for promoting osteoblast adhesion, growth and differentiation was examined. UMR 106 osteoblast-like cells cultured on these surfaces are normal in appearance, and in the presence of serum, proliferate as well or better than on control surfaces. The level of alkaline phosphatase production in the presence and absence of serum suggests that the osteoblast-like cells retain their differentiated phenotype, and appear to respond to the cell surface ligands presented by the osteoblast-membrane biomimetic surface. These observations suggest that biomimetic membrane films prepared from osteoblast cell membranes support osteoblast cell growth, allow the cells to maintain their differentiation state and may be suitable as a model system to probe cell-cell interactions. PMID- 16475848 TI - Functionalisation of surfaces with S-layers. AB - Two-dimensional bacterial surface layer protein crystals (S-layers) are the most commonly observed cell surface structures in prokaryotic organisms (bacteria and archaea). Isolated S-layer proteins have the intrinsic tendency to self-assemble into two-dimensional arrays in suspension and at various interfaces. Basic research on the structure, genetics, chemistry, morphogenesis and function of S layers has led to a broad spectrum of applications in molecular nanotechnology and biomimetics. The possibility to change the natural properties of S-layer proteins by genetic manipulation opens new ways for the tuning of their structural and functional features. Functionalised S-layer proteins that maintain their propensity for self-assembly have led to new affinity matrices, diagnostic tools, vaccines or biocompatible surfaces, as well as to biological templating or specific biomineralisation strategies at surfaces. PMID- 16475849 TI - Biomolecule-compatible support structures for biomolecule coupling to physical measuring principle surfaces. AB - As part of studies on biomolecule-compatible interfacial structures for practice relevant biosensor and biochip developments, new film-forming aminocelluloses of the 'P-CH2-NH-(X)-NH2' type (P = cellulose) with spacer structures (X = special oligoamine residues) at C6 and solubilising groups (S = tosylate or carbanilate) at C2C3 of the anhydroglucose unit (AGU) were synthesised and their film properties and covalent coupling with enzyme protein examined. Depending on the nature and degree of substitution (DS(S)) of the ester groups (S) at C2C3, the new aminocellulose derivatives are soluble either in DMA and DMSO (with S = carbanilate) or in water (with S = tosylate). The aminocellulose derivatives form transparent films from their solutions. AFM investigations of the film surfaces have either shown very flat (topography <1 nm) films or tubular topographies of nanostructure size, depending on structural and environment-induced factors of influence. Especially in the case of films from water-soluble aminocelluloses with oligoamine residues at C6, inter alia, enzyme-specific pH values and different positive charge distributions can be adjusted by partial protonation of the NH2 end groups. By means of the covalent coupling of the new aminocelluloses with glucose oxidase (GOD) it was shown that the enzyme coupling efficiency can be decisively optimised by the interplay of aminocellulose structure, coupling structure and enzyme protein. PMID- 16475850 TI - Ellipsometric microscopy: developments towards biophysics. AB - Ellipsometric microscopy is a novel technique that combines the merits of ellipsometry and light microscopy, i.e. it allows noninvasive, label-free measurements of thin film thickness and refractive index at high lateral resolution. Ellipsometric microscopy has been successfully applied to silicon-air interfaces. However, typical biological systems require immersion in an aqueous buffer. Thus the authors have adapted the instrument for the observation of the interface between glass and water. In particular, the comparatively small differences in refractive indices between substrate and ambient media proved to be a challenge for instrument design. The first experiments with this new instrument are presented. PMID- 16475851 TI - Self-assembling of proteins and enzymes at nanoscale for biodevice applications. AB - Different nanotechnological strategies have been selected to implement biomolecular devices following a bottom-up or top-down approach depending on the biomolecule and on its functionality. Biomolecules have particular functionality and self-assembling capabilities that can be exploited for the implementation of both bioelectronic devices and multipurpose engineered biosurfaces. Surface preparation with supramolecular methods and microcontact printing have been developed and optimised to realise suitable functionalised surfaces. These surfaces can be used to link metalloproteins and enzymes for the implementation of nanobioelectronic devices and planar biosensors or to bind cells in order to promote their growth along predefined tracks and grooves. Some possible applications of these biosurfaces are shown and discussed. Results are presented for the realisation of a biomolecular nanodevice working in air based on the metalloprotein azurin immobilised in the solid state, the formation and characterisation of functional glutamate Dehydrogenase monolayers for nanobiosensing applications, the results of soft lithography processes on azurin for biosensor implementation, and the development of physiological self-assembled patterns of laminin-1 for cell culture applications and hybrid devices. PMID- 16475852 TI - Patterning to enhance activity of cultured neuronal networks. AB - Embryonic rat hippocampal neurons were cultured in order to gain insights into how small networks of neurons interact. The principal observations are the electrical activities recorded with the electrode arrays, primarily action potentials both spontaneous and evoked. Several lithographic techniques were developed for controlling with micrometer precision the patterns of surface molecules in order to control neuronal attachment and growth. Cytophilic polylysine against protein repellent and hence cytophobic polyethylene glycol were used. By combining the cellular lithography with the microelectrode arrays it was possible to guide neurons preferentially to electrodes and to begin to investigate the question as to whether the geometric pattern of a neuronal network influences the patterns of its neuroelectric activity. It is clear that the techniques are adequate to ensure contact of neurons to electrodes but not to ensure the recording of signals, even when neurons lie directly on top of electrodes. The maturation of neuroelectric activity depends on the growth of glia within the culture, such that spontaneous activity appears to become robust when the number of glia is roughly the same as the number of neurons. PMID- 16475853 TI - Pattern modification of a neuronal network for individual-cell-based electrophysiological measurement using photothermal etching of an agarose architecture with a multielectrode array. AB - A new type of individual-cell-based on-chip multielectrode array (MEA) cell cultivation system with an agarose microchamber (AMC) array for topographical control of the network patterns of a living neuronal network has been developed. The advantages of this system are that it allows control of the cell positions and numbers for cultivation using AMCs, as well as easy and flexible control of the pattern of connections between the AMCs through photothermal etching where a portion of the agarose layer is melted with a 1480 nm infrared laser beam. With adequate laser power, narrow micrometer-order grooves (microchannels) can easily be fabricated that can be used to combine neighbouring AMCs to enable topographical control of the neural network pattern. Using this system, an individual-cell-based neural network pattern was formed of rat hippocampal cells within the AMC array without cells escaping from the electrode positions in the microchamber during an eight-day cultivation, and could record cell firing in response to 1.5 V, 500 kHz stimulation through an electrode. This demonstrated the potential of the on-chip AMCMEA cell cultivation system for long-term single cell-based electrophysiological measurement of a neural network system. PMID- 16475854 TI - Analysis of electrotonic coupling in patterned neuronal networks. AB - Microcontact printing of laminin is known as an efficient approach for guiding neuronal cell migration and neurite outgrowth on artificial surfaces. In the present study, ultrathin (approximately 250 microm) brain stem slices of Sprague Dawley rats (E15-E18) were cultured on laminin-patterned substrates, such that neuronal cells migrating out of the slices formed grid-shaped neuronal networks along the geometry defined by the pattern. The interconnections between neighbouring pairs of neurons within these artificial networks were assessed electrophysiologically by double patch-clamp recordings and optically by microinjection of fluorescent dyes. Both functional and electrotonic synapses were detected. Based on the recorded data and simulations in PSpice, an electrical model for electrotonically coupled cells was derived. In this model the neuritic pathway is described as a cylindric cable, and gap junctions are represented by an ohmic resistor. Applying this model in the data analysis, the average inner radius of neurites could be determined to be approximately 0.1 microm. In addition, evidence was found for a correlation between the path-width of the applied pattern and the diameter of neurites growing along these paths. PMID- 16475855 TI - Molecular resolution of cell adhesion forces. AB - Recently, AFM-based force spectroscopy has been used to quantify single-molecule adhesion forces on living ameboid cells. Force spectroscopy was used to measure the rupture forces of single receptor-ligand bonds which can occur rapidly between the cell types used, a metastasising B16 melanoma cell and a vascular bEnd.3 endothelial cell. Parameters which influence the critical experimental conditions are discussed to discriminate between multiple bond ruptures and single bonds. Under physiological conditions of temperature and pH the force measurements show an average rupture force of 33 pN (SD = 12 pN) for single bonds. Single-molecule force spectroscopy will be very useful to study the regulation of cell adhesion on a molecular level in normal processes, such as leukocyte homing, and in major human disorders, including tumor metastasis, autoimmune diseases and atherosclerosis. PMID- 16475857 TI - First steps of an interdisciplinary approach towards miniaturised cryopreservation for cellular nanobiotechnology. AB - The only widely used and accepted method for long-term cell preservation is storage below -130 degrees C. The biosciences will make increasing use of preservation and place new demands on it. Currently, cells are frozen in volumes greater than 1 ml but the new cell and implantation therapies (particularly those using stem cells) will require accurately defined freezing and storage conditions for each single cell. Broadly-based, routine freezing of biological samples allows the advantage of retrospective analysis and the possibility of saving genetic rights. For such applications, one billion is a modest estimation for the number of samples. Current cryotechniques cannot handle so many samples in an efficient and economic way, and the need for new cryotechnology is evident. The interdisciplinary approach presented here should lead to a new sample storage and operating strategy that fulfils the needs mentioned above. Fundamental principles of this new kind of smart sample storage are: (i) miniaturisation; (ii) modularisation; (iii) informationsample integration, i.e. freezing memory chips with samples; and (iv) physical and logical access to samples and information without thawing the samples. In contrast to current sample systems, the prototyped family of intelligent cryosubstrates allows the recovery of single wells (parts) of the substrate without thawing the rest of the sample. The development of intelligent cryosubstrates is linked to developments in high throughput freezing, high packing density storage and minimisation of cytotoxic protective agents. PMID- 16475858 TI - Plasma printing: patterned surface functionalisation and coating at atmospheric pressure. AB - A new plasma-based micropatterning technique, here referred to as plasma printing, combines the well known advantages given by the nonequilibrium character of a dielectric barrier discharge (DBD) and its operation inside small gas volumes with dimension between tens and hundreds of micrometres. The discharge is run at atmospheric pressure and can be easily implemented for patterned surface treatment with applications in biotechnology and microtechnology. In this work the local modification of dielectric substrates, e.g. polymeric films, is addressed with respect to coating and chemical functionalisation, immobilisation of biomolecules and area-selective electroless plating. PMID- 16475859 TI - Transporting cells with mobile microrobots. AB - Current research activities on the development of a system for transport and manipulation of biological cells with microrobots are described. If single cells in liquid are to be placed on a grid or sorted by cell type, having a system that can automatically lift, transport and release cells can significantly speed up such a tedious task. Therefore, a system is being developed that can automatically sort different cells by transporting them to different repositories. A method to recognise different types of cells is also being developed. The system consists of several components; a motorised inverted microscope, several different microrobots and a software architecture to control the whole cell manipulation workstation and to provide a user interface. PMID- 16475860 TI - Microthermoforming as a novel technique for manufacturing scaffolds in tissue engineering (CellChips). AB - The CellChip is a microstructured polymer scaffold, which favours a three dimensional cultivation of cells within an array of cubic microcontainers. The manufacturing process used so far is microinjection moulding combined with laser based perforation. In a first attempt to simplify the process, costly perforation was avoided by using commercially available, inexpensive microfiltration membranes for the bottom of the microcavities. Microthermoforming is a promising novel technique which allows the CellChip to be produced from thin film. Working pressures of approximately 4000 kPa were required for the adequate moulding of 50 microm thick films from three different polymers (polystyrene, polycarbonate, cyclo-olefin polymer). Integrating drafts and chamfers in micromoulds is not going to eliminate an uneven thickness profile, but reduces demoulding forces. Microthermoformed CellChips of polycarbonate were perforated by an ion track technique to guarantee a sufficient supply of medium and gases to the cells. The prestructured CellChips were irradiated with 1460 MeV xenon ions at a fluence of a few 10(6) ions/cm2. The tracks were etched in an aqueous solution of 5 N NaOH at 30 degrees C, which resulted in cylindrical pores approximately 2 microm in diameter. Microinjection-moulded, membrane-bonded and thermoformed CellChips were subjected to comparative examination for viability in a cell culture experiment with parenchymal liver cells (HepG2). The cells stayed viable over a period of more than 20 days. No significant differences in viability between injection moulded, membrane-bonded, and thermoformed CellChips were observed. PMID- 16475861 TI - Microfabricated polymer analysis chip for optical detection. AB - A coupling between multimode polymer waveguides and microfluidic channels on a polymethylmethacrylate (PMMA) capillary electrophoresis (CE)-chip for optical analytical applications has been successfully realised. This technology allows the integration of polymer optical waveguides together with hermetically sealed fluidic channels. The microchannels and waveguides are made in PMMA by the approved hot-embossing technology. The technology developed for the fabrication of polymer waveguides on the microfluidic chip offers the possibility of great flexibility in the choice of core materials, design and alignment of the polymer waveguides. The integration of polymer waveguides on an analysis chip enables highly spatially resolved optical detection without the large and expensive conventionally used apparatus. The optical properties of the analytical system developed are verified by transmission and propagation loss measurements. The results of measurements prove the suitability of the presented device for optical applications between 440 and 800 nm. This was shown with absorbance measurements of the dye Sulfanilazochromotrop (SPADNS) within 50 microm fluidic channels. PMID- 16475862 TI - Microthermoforming of flexible, not-buried hollow microstructures for chip-based life sciences applications. AB - A new method is presented for the manufacturing of flexible, not buried and thin walled hollow microstructures from polymer films. This low-cost method seems to be especially suited for the fabrication of plastic microstructures for fluidic one-way applications in the field of life sciences. It is based on a thermoforming process adapted to microstructure technology and is called 'microthermoforming'. Inside a hot embossing press, a heated thin thermoplastic film is formed into the evacuated microcavities of a plate-shaped metal mould using a compressed gas. The film may be heat-sealed on to a thicker plastic film substrate inside the same press without demoulding the thermoformed film. To demonstrate the performance of the new manufacturing method, flexible capillary electrophoresis and cell culture chips from polystyrene, polycarbonate and a cyclo-olefin polymer with 16 and 625 parallel microstructures each, respectively, have been fabricated. PMID- 16475863 TI - From micro to nano: properties and potential applications of micro- and nano filled polymer ceramic composites in microsystem technology. AB - In microsystem technology, four important material classes are established either for the generation or the replication of microstructured surfaces: silicon, polymers, metals and ceramics. Composite materials consisting of a polymer matrix and ceramic fillers show improved thermomechanical properties in comparison to polymers and can be introduced as a new additional material class. The substitution of micro-sized ceramic fillers by nano-sized ceramics in composites has a strong influence on the composite's physical properties: the reduction of ceramic particle size down to the nanometre scale results in an improved sinter activity owing to the large surface area. The fabrication of dense ceramics is simplified and can be used for a rapid prototyping of microstructured ceramic parts. The addition of nano-sized ceramics with particle sizes of <40 nm to polymers allows the manufacturing of transparent polymer based composites with modified refractive indices for use in polymer waveguides. The influence of the ceramic particle size, the ceramic content and different dispersion methods on the composite's physical properties are discussed. PMID- 16475864 TI - Ambipolar transistors based on azurin proteins. AB - A new type of transistor is presented. It is realised by using a metalloprotein; namely, azurin. Thanks to their natural functional characteristics, which involve inter- and intramolecular electron transfer, metalloproteins are good candidates for biomolecular nanoelectronics. The implementation of a prototype of protein transistor operating in air and in the solid state based on self-organised films of azurins is reported. Experimental current-voltage characteristics are shown. The new device presents an ambipolar behaviour as the gate bias voltage is changed. Exploiting this peculiar characteristic, a fully integrated logic gate which can be a good starting point for a new class of nanoelectronics devices has been realised. PMID- 16475865 TI - Preparation and purification of synthetic protein nanoparticulates. AB - The protein nanostructure used in this study (bovine serum albumin; BSA nanoparticles) were fabricated with an average nanoparticle diameter 150 nm based on the principle of coacervation. Practical recovery of nanoparticulate mimics, of products such as plasmid DNA and viruses as putative gene therapy vectors from model systems, has been studied. The adsorbents employed in this study for the recovery of nanoparticles had one of four discrete designs i.e. microporous (pore size <0.2 microm), macroporous (pore size >0.8 microm), solid phase (nonporous) and pellicular (pore size <0.5 microm). Soluble protein was included in the study to represent cellular components of complex feedstocks and the separation of assemblies from components, while particulate protein served as surrogate size and charge mimics of less easily sourced viral and plasmid gene therapy vectors. Candidate adsorbents were physically characterised to assess their suitability for fluidised-bed operation, biochemically characterised exploiting batch-binding experimentation and laser scanning confocal microscopy. The adsorptive capacity of nanoparticulate products was strongly influenced by the physical design of the adsorbents, and microporous adsorbents appeared to be less suited for the recovery of nanoparticulate products. The generic application of such adsorbents for the recovery of nanoparticulate bioproducts is discussed. PMID- 16475866 TI - Principles of a nanoscale DNA scanner. AB - A nanofluidic device for the routine stretching of single DNA molecules was hypothesised and tested. The device consists of an entrance channel leading to a post-field preceding an elongational flow field. The device facilitates each molecule's coil-to-stretch transition, counteracts its entropic recoil, and presents a stream of moving stretched molecules for detailed single-molecule time of-flight measurements. The physics of DNA stretching was explored in a device where there was a juxtaposition of hexagonal upright post arrays with lithographically defined elongational flow fields. PMID- 16475867 TI - Tunable optical properties of colloidal quantum dots in electrolytic environments. AB - The absorption spectra of colloidal cadmium sulfide quantum dots in electrolytic solutions are found to manifest a shift in the absorption threshold as the concentration of the electrolyte is varied. These results are consistent with a shift in the absorption threshold that would be caused by electrolytic screening of the field caused by the intrinsic spontaneous polarisation of these wurtzite structured quantum dots. These electrolyte-dependent absorption properties provide a potential means of gaining insights on the variable extracellular and intracellular electrolytic concentrations that are present in biological systems. PMID- 16475868 TI - Modelling current transport through DNA (deoxyribonucleic acid) molecules using equivalent circuits. AB - The current transport properties of DNA molecules are of considerable interest. The key reason for this appears to be linked to the universality of DNA molecules in living organisms, their self-assembly properties, and potential applications as nanoscale devices. The modelling of the I-V characteristics of a DNA molecule using equivalent circuits is reported. The advantages of the proposed model are that non-linear current behaviour can be included together with potential piece wise solutions. The model includes the use of transistors to mimic current discontinuities at transition points. The simulated results closely resemble measured I-V curves and do not invoke resonant tunneling which contradicts observed temperature dependences. An equivalent-circuit model which includes the use of active devices is shown to be effective way to mimic non-linear current transport in biological molecules. PMID- 16475869 TI - A stress surveillance system based on calcium and nitric oxide in marine diatoms. AB - Diatoms are an important group of eukaryotic phytoplankton, responsible for about 20% of global primary productivity. Study of the functional role of chemical signaling within phytoplankton assemblages is still in its infancy although recent reports in diatoms suggest the existence of chemical-based defense strategies. Here, we demonstrate how the accurate perception of diatom-derived reactive aldehydes can determine cell fate in diatoms. In particular, the aldehyde (2E,4E/Z)-decadienal (DD) can trigger intracellular calcium transients and the generation of nitric oxide (NO) by a calcium-dependent NO synthase-like activity, which results in cell death. However, pretreatment of cells with sublethal doses of aldehyde can induce resistance to subsequent lethal doses, which is reflected in an altered calcium signature and kinetics of NO production. We also present evidence for a DD-derived NO-based intercellular signaling system for the perception of stressed bystander cells. Based on these findings, we propose the existence of a sophisticated stress surveillance system in diatoms, which has important implications for understanding the cellular mechanisms responsible for acclimation versus death during phytoplankton bloom successions. PMID- 16475870 TI - Topical bioadhesive patch systems enhance selectivity of protoporphyrin IX accumulation. AB - In clinical 5-aminolevulinic acid (ALA)-based photodynamic therapy (PDT) of skin tumors it is desirable to develop vehicles that minimize the penetration of ALA through normal stratum corneum and maximize it through the compromised stratum corneum of the tumors to improve tumor selectivity. We have designed a bioadhesive patch, which may be able to achieve this aim. It induces levels of protoporphyrin IX (PpIX) in skin overlying tumors similar to those induced by the proprietary cream (Porphin) but at the same time induces less PpIX to form in normal skin and at distant sites. The mechanisms of action of the patch, as compared with that of the cream, were studied by means of Cuprophan barriers that mimic compromised tumor stratum corneum and in a mouse model with transplanted tumors. PMID- 16475871 TI - Quantum dot-based energy transfer: perspectives and potential for applications in photodynamic therapy. AB - Quantum dots have emerged as an important class of material that offers great promise to a diverse range of applications ranging from energy conversion to biomedicine. Here, we review the potential of using quantum dots and quantum dot conjugates as sensitizers for photodynamic therapy (PDT). The photophysics of singlet oxygen generation in relation to quantum dot-based energy transfer is discussed and the possibility of using quantum dots as photosensitizer in PDT is assessed, including their current limitations to applications in biological systems. The biggest advantage of quantum dots over molecular photosensitizers that comes into perspective is their tunable optical properties and surface chemistries. Recent developments in the preparation and photophysical characterization of quantum dot energy transfer processes are also presented in this review, to provide insights on the future direction of quantum dot-based photosensitization studies from the viewpoint of our ongoing research. PMID- 16475873 TI - Home medication reviews: what do people in the Australian Riverland think of them? PMID- 16475872 TI - Large-scale evidence for the effect of the COLIA1 Sp1 polymorphism on osteoporosis outcomes: the GENOMOS study. AB - BACKGROUND: Osteoporosis and fracture risk are considered to be under genetic control. Extensive work is being performed to identify the exact genetic variants that determine this risk. Previous work has suggested that a G/T polymorphism affecting an Sp1 binding site in the COLIA1 gene is a genetic marker for low bone mineral density (BMD) and osteoporotic fracture, but there have been no very large-scale studies of COLIA1 alleles in relation to these phenotypes. METHODS AND FINDINGS: Here we evaluated the role of COLIA1 Sp1 alleles as a predictor of BMD and fracture in a multicenter study involving 20,786 individuals from several European countries. At the femoral neck, the average (95% confidence interval [CI]) BMD values were 25 mg/cm2 (CI, 16 to 34 mg/cm2) lower in TT homozygotes than the other genotype groups (p < 0.001), and a similar difference was observed at the lumbar spine; 21 mg/cm2 (CI, 1 to 42 mg/cm2), (p = 0.039). These associations were unaltered after adjustment for potential confounding factors. There was no association with fracture overall (odds ratio [OR] = 1.01 [CI, 0.95 to 1.08]) in either unadjusted or adjusted analyses, but there was a non significant trend for association with vertebral fracture and a nominally significant association with incident vertebral fractures in females (OR = 1.33 [CI, 1.00 to 1.77]) that was independent of BMD, and unaltered in adjusted analyses. CONCLUSIONS: Allowing for the inevitable heterogeneity between participating teams, this study-which to our knowledge is the largest ever performed in the field of osteoporosis genetics for a single gene-demonstrates that the COLIA1 Sp1 polymorphism is associated with reduced BMD and could predispose to incident vertebral fractures in women, independent of BMD. The associations we observed were modest however, demonstrating the importance of conducting studies that are adequately powered to detect and quantify the effects of common genetic variants on complex diseases. PMID- 16475874 TI - Development of an integrated and sustainable rural service for people with diabetes in the Scottish Highlands. AB - INTRODUCTION: The number of people with diabetes is increasing leading to a greater burden on health care services. The impact of the growing prevalence is accentuated by remote and rural demographic and geographic characteristics. Highland is a sparsely populated remote and rural area in the north of Scotland, characterised by poor access to health-care services and pockets of marked deprivation. Centralised policy developments demanding local implementation compounded the pressures on a system that already had waiting times of over 90 weeks for some people with diabetes. A regional review of services, engaging stakeholders from all disciplines and geographical locations was required to develop acceptable and sustainable solutions. This article describes the extensive mapping process involved, how solutions were derived, and suggests a new service structure to encompass remote health-care issues. METHODS: Health care professionals with an interest in diabetes were identified and workshops were organised to include the remote areas of Highland. Patient and carers views were ascertained through workshops and supplemented by written submissions. Using the redesign methodology the patient pathway was mapped, noting service deficiencies and good practice. The information gathered was constructed into a service-level map representing the patient journey. A conference was organised to develop solutions to the issues raised during the mapping process. From these solutions a new service configuration was constructed. RESULTS: Over 300 health care professionals patients and carers contributed. Fourteen workshops were held across the region including the remote areas, providing 15 local maps of the patient pathways subsequently amalgamated into a service-level map. The current patient pathway in Highland follows a traditional and dichotomous cycle of care in the primary and secondary care setting, partly reflecting the rural nature of healthcare in the Highlands. Four main areas for service improvement were identified: a reduction in waiting times for secondary care out patients; an improvement in communication between health-care professionals; further education for both health-care professionals and patients; and the use and role of allied health professionals. Seventeen solutions were recommended, including the introduction of a managed clinical network, use of an integrated IT system, use of a remote access consultation clinic, and web-based peer education and group sessions for allied health professionals. A new service configuration was proposed with the patient at the centre of a non-hierarchical system using standardised referral letters with a seamless flow of information. CONCLUSION: Local processes for the implementation of government directives are imposing pressures on relatively smaller organisations. These pressures develop as a result of attempts to ensure local ownership and in overcoming the difficulties unique to the remote and rural setting. Further evaluation of the implementation of initiatives to solve the issues of service planning in remote areas is needed to clarify their level of effectiveness. PMID- 16475875 TI - Correlation of heavy and light chain mRNA copy numbers to antibody productivity in mouse myeloma production cell lines. AB - Manufacturing cell line development at Centocor involves transfection of antibody genes into host cell lines and isolating primary transfectomas that upon subcloning yield high expressing cell lines for the desired antibody. In an attempt to increase productivity of these cell lines, we set out to identify the rate-limiting step in the process of antibody expression and secretion. For this purpose, 30 antibody expressing cell lines with variable antibody expression levels were analyzed for heavy-chain and light-chain mRNA expression levels. Results suggested that the increase in antibody titer of the subclones (compared to their primary clones) was partly due to an increase in heavy-chain and light chain mRNA levels; higher expressers were associated with approximately 1.0 x 10(7) and 1.5 x 10(7) copies of heavy-chain and light-chain per 10 nanogram of cDNA, respectively. Generally, the level of light-chain mRNA was higher compared to the level of heavy-chain mRNA in a majority of the cell lines, and the difference in their levels was not due to their differential stability. The data generated from all the cell lines tested in this study suggested that there was a correlation of light-chain and heavy-chain transcript levels to antibody productivity, with the coefficient of correlation being 0.59 for light chain and 0.81 for heavy chain. We conclude that transcription of heavy chain and to a lesser extent light chain could be one of the rate-limiting steps in the antibody expression pathway. Hence, methods that would increase these mRNA levels could be beneficial in the attempt to improve the antibody expression level of production cell lines. PMID- 16475876 TI - Canine perianal gland carcinoma-associated antigens defined by monoclonal antibodies. AB - This study was conducted to distinguish canine perianal gland carcinomas from adenomas using monoclonal antibodies (MAbs). The adenomas generally retain the lobular architecture, but some may contain focal areas of cellular pleomorphism. These changes may suggest malignant transformation and have led to discordant interpretations. To address this histopathological confusion, two perianal gland carcinoma-associated antigens were defined by mouse MAbs 4A9 and 1A10. These MAbs, generated against a canine mammary carcinoma cell line, reacted strongly with perianal gland carcinoma in preliminary screening and therefore were selected for further investigation. Cellular expression of antigens was examined by indirect immunoperoxidase (IP) assay using MAbs 4A9 and 1A10 against formalin fixed, paraffin-embedded sections of normal and tumor tissue. Of 25 perianal gland carcinomas, 4A9 antigen was expressed in 100% and 1A10 antigen in 84%. In contrast, perianal gland adenomas were negative for both antigens, and little or no reactivity was detected with normal perianal glands. With eight perianal gland tumors, diagnosis of carcinoma versus adenoma was histologically equivocal, while IP assays consistently revealed focal expression of the 4A9 and 1A10 antigens in these tumors, and the staining coincided with foci of anaplastic cells having a high mitotic index. This group of tumors was designated adenoma/carcinoma in situ. Results suggest that 4A9 and 1A10 antigens are markers of carcinoma and malignant transformation in canine perianal gland tumors, and can be very useful as diagnostic reagents where the identification of carcinoma versus adenoma requires additional clarification beyond routine histopathological examination. PMID- 16475877 TI - Preparation of tissue-specific monoclonal antibodies using purified endothelial membrane proteins from biotinylated pulmonary vasculature of rhesus monkey. AB - In the present study, we perfused the rhesus lung vascular bed in situ with sulfo NHS-LC-Biotin to biotinylate its luminal surface membrane proteins. After homogenization, dialysis, and affinity chromatography, biotinylated endothelial membrane proteins were successfully isolated and characterized as enriched endothelial membrane proteins with no contamination of intracellular proteins. When they were used as immunogens to develop monoclonal antibodies (MAbs), three MAbs--TX111, TX112, and TX113--were obtained. Among them, TX111 was demonstrated to specifically bind to rhesus lung tissue by Western blotting and enzymelinked immunosorbent assay (ELISA)--that is, positively stained capillary endothelium of rhesus lung. The molecular weight of the corresponding antigen for TX111 was approximately 70 kDa under reducing conditions. TX111 also reacted with human lung homogenate, but not with rat lung homogenate. These results suggest that (1) the biotinylation method is applicable for isolating endothelial proteins in situ from large animals; (2) anti-human protein MAbs are likely to be obtained using monkey proteins; and (3) TX111 is potentially useful for pulmonary vascular targeting. PMID- 16475878 TI - Preparation and characterization of monoclonal antibody against protein TREM-like transcript-1 (TLT-1). AB - TREM-like transcript-1 (TLT-1) was initially characterized as the putative inhibitory receptor within the TREM locus on chromosome 6 and was suggested to play a critical role in modulating TREM-associated immunity. Recently, TLT-1 was found not to function as an inhibiting member of the TREM family; however, it may play a significant role in maintaining vascular hemostasis, coagulation, and inflammation at sites of injury through its location in platelets. In this study, we have successfully generated a monoclonal antibody that can be applied to assays, including enzyme-linked immunosorbent assay (ELISA), Western immunoblotting, immunofluorescence, and Fluorescence-Activated Cell Sorter (Becton-Dickinson) (FACS), for the study of TLT-1 functions in physiological and pathological conditions. PMID- 16475879 TI - Production of monoclonal antibodies against Prox1. AB - Prox1 is a divergent homeodomain protein important for the development of the lens, retina, liver, pancreas, and lymphatic vasculature. Prox1 expression is highly upregulated in transformed hepatocytes and has been used as a marker to distinguish lymphatic from blood vasculature. We produced recombinant human Prox1 (amino acids 547-737) fused to glutathione S-transferase (GST) and used it to create two hybridomas, 5G10 and 4G10. Both of these hybridomas produced monoclonal antibodies able to detect Prox1 by immunofluorescence in lenses from diverse terrestrial vertebrates, including humans, rats, chickens, and lizards, although 5G10 was generally more sensitive in this application. Further, 4G10 was able to robustly detect endogenous and recombinant Prox1 in both cell and tissue extracts by Western blotting, while 5G10 was notably less sensitive for this purpose. These monoclonal antibodies will be useful for diverse studies on the role of Prox1 in both normal development and disease processes in terrestrial vertebrates. PMID- 16475880 TI - Production and characterization of monoclonal antibodies against 17alpha hydroxyprogesterone. AB - Hybridomas secreting monoclonal antibodies (MAbs) against 17alpha hydroxyprogesterone (17OHP) have been generated. These MAbs are highly specific and have an affinity of 7-12 x 10(7) M(1). The hybridomas were obtained by fusion of spleen cells from immunized mice with mouse myeloma P3X63 Ag8.653 cells. The antigen used for immunization was 17OHP conjugated to bovine serum albumin (17OHP:BSA). Fused cells were plated and cloned in 96-well microtiter plates. Wells containing hybridomas were screened simultaneously for specific gamma globulin (IgG) and anti-17OHP activity using an enzyme-linked immunosorbent assay (ELISA)-based method, which is faster than the conventional radioimmunoassay (RIA) screening procedure. Limiting dilution methods were used to obtain single hybridoma clones producing MAb. The stable hybridomas secreting anti-17OHP MAbs were expanded into bioreactors or ascites fluid for large-scale production of the required antibodies. These MAbs will be used in the formulation of a 17OHP assay kit to screen for congenital adrenal hyperplasia (CAH) in local newborn human population. PMID- 16475887 TI - Erectile dysfunction in an urban HIV-positive population. AB - The International Index of Erectile Function (five-item version) was used to assess the prevalence and risk factors of erectile dysfunction (ED) in an HIV positive urban population of men. The prevalence of ED was found to be 74%. Age, CD4 count, history of opportunistic infections, highly active antiretroviral therapy (HAART) use, and serum testosterone level were not significantly associated with ED in this study. These results emphasize the high prevalence of ED in HIV-positive men. PMID- 16475881 TI - Generation and characterization of an hKid-specific monoclonal antibody. AB - Chromokinesins are chromosome-bound proteins during mitosis that play multiple important roles in chromosome segregation. The chromokinesin Kid has been shown to be involved in chromosome congression during mitosis and meiosis. Here we have generated a monoclonal antibody specific for the human chromokinesin hKid by immunizing BALB/c mice with a recombinant protein fragment corresponding to the C terminal 250-amino acid residues of hKid. All five immunized mice responded excellently and gave high, nearly monospecific, antibody titers. One hybridoma, 8C12, was generated from spleen cells of a selected mouse, which recognized hKid on immunoblots and in immunofluorescence experiments. As hKid is an important regulator of chromosome segregation, this monoclonal antibody will be a useful tool for further analysis of this chromokinesin. PMID- 16475888 TI - Routine testing for HIV in the United States: the intersection between recommendations and practice. AB - The purpose of this study was to examine the current practices of family practice (FP) providers and their allied staff with regard to routine HIV testing in Rhode Island (RI) and Mississippi (MS). Anonymous experimenter-derived surveys were mailed to both groups of providers in 2002. The questionnaire contained five questions about their current practices and attitudes toward HIV testing as well as patient demographics. Five hundred twenty-one questionnaires were sent to American Academy of Family Practitioners (AAFP) members in RI and MS and to FPs with listings in the phone book in RI. The response rate was 52% in RI and 41% in MS. The vast majority of providers (93%) tested their high-risk patients for HIV, but less tested pregnant (57%) and other sexually active (37%) patients. The FPs in this survey wanted HIV testing to be done in the primary care setting, yet only 7% recommended HIV testing to their sexually active patients aged 18-50 in the previous year. In order not to stigmatize any specific risk group, nor to miss any patients who are unable to be identified as being "at high risk," routine testing in the primary care setting should be encouraged. PMID- 16475889 TI - Refusing HIV testing in an urgent care setting: results from the "Think HIV" program. AB - Efforts to increase HIV case identification through routine, voluntary HIV testing are hindered by high refusal rates. Our objective was to identify patients most likely to refuse routine HIV testing. We developed a new HIV testing program at four Massachusetts urgent care centers. Patients were asked if they were interested in routine HIV testing. We performed analyses to assess differences in characteristics between those who refused testing and those who accepted it. Data were available for 9129/10,354 (88%) patients offered routine HIV testing from January to December 2002. Of these 9129 patients, 67% refused testing. In the crude analysis, HIV test refusal was associated with female gender, white race, older age, and higher educational level. In multivariate analysis, non-English-speaking patients who were Hispanic, Haitian, and other race were more likely to refuse testing than their English-speaking counterparts. Among all patients, "not at risk" and "already tested" were the most common reasons for test refusal. Two thirds of patients refused routine HIV testing when it was offered in a statewide urgent care-based program. If routine HIV testing programs are to be successful, strategies must be developed to increase HIV test acceptance among patients most likely to refrain from testing. PMID- 16475890 TI - HIV in correctional facilities: role of self-report in case identification. AB - For treatment of HIV/AIDS in jails and prisons to be effective, these institutions must identify as many HIV-positive inmates as they can. We compare HIV status among a drug-addicted jail population determined through a physical examination and a voluntary HIV testing program, with self-reported status in an interview. Of 360 subjects interviewed and given physical examinations, approximately one third (110) took the voluntary HIV test and all were negative, and only 1 was identified as HIV-positive in the physical. However, 7 (2%) stated in the interview that they were HIV positive, none of whom took the HIV test. Five of the 7 also self-reported injection drug use and having shared needles. We conclude that inmate self-report is an important pathway for HIV case finding in correctional institutions. PMID- 16475891 TI - Religious beliefs, practices and treatment adherence among individuals with HIV in the southern United States. AB - Nonadherence with medical treatment is a critical threat to the health of those living with HIV disease. Unfortunately the search for explanatory factors for nonadherence is still not fully developed, particularly in the area of religion and spirituality. Extant literature suggests that church attendance, religious practices and spiritual beliefs may improve health and generally benefit patients. However, religious beliefs may also play a negative role in treatment adherence due to the stigma attached to HIV disease, particularly in geographical areas and in population subgroups where religious practices are strong. In this exploratory study, HIV-positive individuals (n = 306) in a southern state were surveyed as to their attitudes and beliefs surrounding HIV disease and adherence with medical treatment for the disease. The results indicate that multiple factors influence adherence with treatment and that certain religious practices are positively associated with adherence, but certain religious beliefs are negatively related to adherence. The findings of this study reinforce the importance of remembering and addressing a patient's religious beliefs as a part of medical care. PMID- 16475892 TI - Use of multisystemic therapy to improve antiretroviral adherence and health outcomes in HIV-infected pediatric patients: evaluation of a pilot program. AB - The purpose of the current study was to evaluate a clinical program that used multisystemic therapy (MST) to improve regimen adherence and health outcomes among children with perinatally acquired HIV who exhibited high viral loads in the absence of viral resistance. MST is an intensive, home-based family therapy previously used for the treatment of serious antisocial behavior. Goals of the study were to assess the feasibility of the MST program and its effectiveness for improving health outcomes among children who were treated. The study was conducted by retrospective chart review. Ninety percent of children and families referred to the program accepted the referral and 95% received a full dose of treatment, suggesting high program feasibility. Nineteen children participated in the program. General HIV knowledge on the part of caregivers improved significantly over the course of treatment. Although caregiver-reported adherence did not change, viral loads were found to significantly decrease from referral to the end of MST treatment, with the mean change reflecting a greater than 1 log10 decrease. The majority of children maintained these improvements during the 3 months after treatment termination. Results suggest that MST holds promise as an intervention for improving health outcomes among pediatric patients with HIV. PMID- 16475893 TI - Domestic violence in barriers to health care for HIV-positive women. AB - Few studies have addressed the issue of domestic violence and health care for HIV positive women. However, such women are at increased risk of clinical progression when domestic violence prevents access to health care or their ability to take HIV medicines on a consistent basis. To address this issue, 3 focus groups and 50 in-depth interviews were conducted at a public health clinic with HIV-positive women clients who had experienced domestic violence. The results are illustrated in 4 case studies of how domestic violence diminishes women's ability to obtain regular health care. Abused women were reluctant to keep appointments if they were afraid of their partners, if they were depressed, feeling ill or "too worn down," or if they were ashamed of being abused. Abusive partners were sometimes reported to sabotage women's efforts to seek care, keep appointments or take medications. The study concluded that domestic violence is an underrecognized barrier to women's ability to obtain regular medical care for HIV/AIDS. Effective HIV treatment is dependent on consistent HIV care, and domestic violence is a crucial barrier for some women. More research is needed to determine the most effective interventions for domestic violence in relation to HIV-positive women. PMID- 16475894 TI - Understanding interrelationships among HIV-related stigma, concern about HIV infection, and intent to disclose HIV serostatus: a pretest-posttest study in a rural area of eastern China. AB - The objective of the study was to examine the interrelationships among HIV related public and felt stigma, worry of HIV infection, HIV/AIDS knowledge and intention to disclose HIV testing results in a rural area of China, where HIV spread among former commercial blood donors. A one-group pretest-posttest study was conducted among 605 marriage license applicants. The following relationships showed statistical significance in path analysis: (1) HIV/AIDS knowledge --> worry [beta (Standardized coefficient) = -0.39]; (2) worry --> public stigma (beta = 0.27); (3) public stigma --> felt stigma (beta = 0.22); and (4) felt stigma --> intention to disclosure (beta = -0.20). Separate path analyses for males and females generated similar association patterns. HIV counseling reduced perceived worry but exerted little impact on HIV-related stigma and the intention. The pathway from a lack of HIV/AIDS knowledge to increased stigma and to decreased intention to disclose one's serostatus is particularly policy relevant as decreased intention to disclosure may be related to continuing practice of HIV risk behaviors. The findings demonstrate interventions aiming at the reduction of stigma should be targeted at both the individual and community levels. PMID- 16475895 TI - Allergy testing: the role of recombinant allergens. AB - Currently, diagnosis of type I allergy is performed using crude allergen extracts, which allow the identification of the allergen-containing source responsible for type I allergic symptoms (e.g., allergic rhino-conjunctivitis, asthma) but not the disease-eliciting molecules. With the introduction of recombinant allergens produced by molecular biology techniques, a large panel of allergenic molecules has become available. The application of these recombinant allergens for in vitro tests has led to new forms of component-resolved diagnosis (CRD) and allows the establishment of a patient's individual reactivity profile. The increasing number of recombinant allergens characterized during the last decade has allowed the development of chip-based allergy tests for simultaneous detection of up to 5000 different allergens and epitopes. The introduction of these recombinant allergen-based tests into clinical practice improves the selection of patients for traditional specific immunotherapy and allows monitoring of the immunological efficacy of specific immunotherapy by measuring allergen-specific IgG antibodies. Besides their diagnostic application, recombinant allergens and hypoallergenic derivatives thereof have also been used as vaccines in clinical trials, and recent results have shown their usefulness for the treatment of type I allergy. PMID- 16475896 TI - Autoimmune diagnostics in diabetes mellitus. AB - Type 1 diabetes results from a specific destruction of the insulin-producing beta cells of the pancreas. The disease is characterized by the appearance of specific autoantibodies against islet cell antigens. Autoantibodies to insulin, glutamic acid decarboxylase, tyrosine phosphatase IA-2 and cytoplasmic islet cell antibodies are useful markers for the differential diagnosis of type 1 diabetes when clinical and metabolic criteria alone do not allow definite classification. Autoimmune diagnostics is of particular importance in adults to discriminate between type 1 and type 2 diabetes and to assess the diagnosis of latent autoimmune diabetes in adults. PMID- 16475897 TI - Diagnosis and prognosis of early rheumatoid arthritis, with special emphasis on laboratory analysis. AB - Diagnosis of rheumatoid arthritis (RA) is mainly based on clinical criteria of symmetric polyarthritis of the hands and feet, with morning stiffness lasting usually more than 1 h. Autoantibodies typical for RA, i.e., rheumatoid factors and anti-cyclic citrullinated peptide, and measurements of inflammation add more specific information, especially for early diagnosis, where clinical presentation may be oligosymptomatic involving only a few joints. These laboratory parameters are also relevant for prognosis of disease persistence, functional impairment and radiological progression. PMID- 16475898 TI - Autoimmune bullous disorders. AB - Bullous skin diseases represent a group of organ-specific autoimmune disorders characterised by binding of circulating autoantibodies to adhesion molecules of the epidermis and the dermo-epidermal basement membrane zone. Binding of these autoantibodies to their antigenic targets results in loss of adhesion between epidermal keratinocytes and at the level of the basement membrane zone. Chronic blisters and secondary painful erosions are the clinical hallmark of autoimmune bullous disorders. Histopathology reveals the location of blister formation and helps to classify the subtype of the bullous skin disorder. Immunofluorescence is crucial for diagnosing autoimmune bullous skin disorders. Tissue-bound autoantibodies are detected by direct immunofluorescence of perilesional skin. Circulating autoantibodies can be visualised by indirect immunofluorescence using tissue substrates such as monkey oesophagus and sodium chloride-split human skin. Most of the autoantigens are available as recombinant proteins, which allows for autoantibody screening by ELISA or immunoblot analysis to confirm the primary diagnosis and, importantly, for immunoserological follow-up of patients. PMID- 16475899 TI - Laboratory network of excellence: enhancing patient safety and service effectiveness. AB - Clinical laboratories have undergone major changes due to technological progress and economic pressure. While costs of laboratory testing continue to be the dominant issue within the healthcare service worldwide, quality, effectiveness and impact on outcomes are also emerging as critical value-added features. Five Italian laboratories are therefore promoting a network of excellence by investigating markers of effectiveness of laboratory services and sharing their experience of using them in clinical practice. In the present study we report preliminary data on indicators of quality in all phases of the so-called total testing process, the key to evaluating all phases of the total testing process, including the appropriateness of test requests and data interpretation. Initial findings in evaluating pre-analytical causes of specimen rejection in three different laboratories and the effects of introducing three laboratory clinical guidelines are reported. These data should stimulate debate in the scientific community and encourage more clinical laboratories to use the same indicators to improve clinical effectiveness and clinical outcomes within the healthcare service. PMID- 16475900 TI - Novel mutations in the 5'-UTR of the FOLR1 gene. AB - We have previously reported two novel mutations in the 5'-untranslated region (UTR) of the gene for folate receptor-alpha (FOLR1). In our search for additional mutations, 92 patient samples with elevated levels of homocysteine were screened by single-strand conformation polymorphism (SSCP) between nt -425 and -782, and 712 and -1110. Between nt -425 and -782 we did not find any mutations. Between nt -712 and -1110 there were three novel mutations. One subject had two mutations very close to each other, c.-856C>T and c.-921T>C. Two subjects had a c.-1043G>A mutation. To get an idea of the prevalence of FOLR1 mutations in an unselected population, we also screened 692 healthy school children for mutations. In this cohort, between nt -188 and +272 we discovered one novel mutation, a single nucleotide substitution, c.-18C>T, in addition to five children with the 25-bp deletion mutation previously described by us. Thus, so far we have discovered six novel mutations in the 5'-UTR region of the gene for folate receptor-alpha. We genotyped all 17 subjects with a FOLR1 mutation for the methylenetetrahydrofolate reductase (MTHFR) 677C>T polymorphism, and developed new single-nucleotide polymorphism (SNP) genotyping protocols for MTHFR 1298A>C and 1793G>A utilising Pyrosequencing technology. None of the 17 subjects had the 677TT genotype, which ruled out this as a cause of elevated homocysteine levels, which was observed in some of the subjects. Further studies of mutations in the 5'-UTR of FOLR1, and in particular of their interplay with folate intake status, are warranted. PMID- 16475901 TI - CYP2C8 polymorphism among the Portuguese. AB - Cytochrome P450 2C8 (CYP2C8) is a polymorphic phase I drug-metabolising enzyme involved in the metabolism of a wide variety of xenobiotics, as well as a proposed player in the regulation of vascular tone. Polymorphisms in this gene may have an impact on the metabolism of therapeutic drugs such as paclitaxel and verapamil. In this report we have determined the frequencies of the main non synonymous CYP2C8 alleles, 805A>T (CYP2C8*2), 416G>A/1196A>G (CYP2C8*3) and 792C>G (CYP2C8*4) in a sample representative of Portuguese Caucasians. The allelic frequencies determined were 1.2%, 19.8%, and 6.4% for CYP2C8*2, CYP2C8*3, and CYP2C*4, respectively. The observed CYP2C8*3 prevalence is significantly different from the frequencies previously reported in North European populations. PMID- 16475902 TI - Actual levels of soy phytoestrogens in children correlate with thyroid laboratory parameters. AB - Thyroid hormones and thyroid autoantibodies, along with serum concentrations of two phytoestrogens of the isoflavone series, daidzein and genistein, were measured in 268 children without overt thyroid diseases, screened for iodine deficiency in one region of the Czech Republic. Since both phytoestrogens have been reported to inhibit thyroid hormone biosynthesis and in high concentrations to exert goitrogenic effects, we investigated whether their presence in the circulation could influence thyroid hormone function in a population where soy consumption is not common. Correlation analysis revealed a significant positive association of genistein with thyroglobulin autoantibodies and a negative correlation with thyroid volume. Multiple regression analysis of the relationships between actual phytoestrogen levels and measured thyroid parameters revealed only a weak but significant association between genistein and thyroid variables. Higher levels of free thyroxine were found in a subgroup of 36 children who ate soy food in the previous 24 h. In conclusion, only modest association was found between actual phytoestrogen levels and parameters of thyroid function. On the other hand, even small differences in soy phytoestrogen intake may influence thyroid function, which could be important when iodine intake is insufficient. PMID- 16475904 TI - Comparison of Brucella immunoglobulin M and G flow assays with serum agglutination and 2-mercaptoethanol tests in the diagnosis of brucellosis. AB - The diagnostic value of Brucella IgM/IgG flow assays was evaluated in comparison with serum agglutination and 2-mercaptoethanol tests by testing a selection of serum samples submitted to the laboratory because of clinical suspicion of brucellosis. All 39 admission and 11 follow-up samples that agglutinated in the serum agglutination test tested positive in the flow assay, whereas all 20 serum agglutination negative samples with clinical suspicion of brucellosis, 23 control samples from healthy individuals and 20 control samples from cases with chronic hepatitis tested negative in the flow assay. The Brucella IgM and IgG flow assays were slightly more sensitive than the agglutination tests in discriminating between specific IgM and IgG antibodies. The Brucella IgM and IgG flow assays are easy-to-perform and quick assays that can be used for the diagnosis of brucellosis. The flow assays are very useful, especially in rural settings where brucellosis is widespread and where well-equipped laboratories to perform the laboratory tests are not readily available. PMID- 16475903 TI - Comparison of the concentration of trace metals (Ni, Zn, Co, Cu and Se), Fe, vitamins A, C and E, and lipid peroxidation in patients with prostate cancer. AB - The anticarcinogenic and antioxidant properties of vitamins A, C, E and pro- or antioxidant properties of trace metals have recently attracted increased attention. We examined the levels of antioxidant vitamins (A, C and E), selenium and malondialdehyde (MDA), and trace metals (Fe, Ni, Zn, Co and Cu) in patients with prostate cancer. In total, 41 subjects (21 controls and 20 prostate cancer patients) were included in the study. The levels of trace elements and Fe in whole blood were determined by atomic absorption spectrophotometry. Serum levels of Se were determined using a fluorimetric method, while a HPLC method was used for serum levels of vitamins and MDA. The levels of vitamins A and E were significantly lower and MDA levels were significantly higher (p<0.001) in patients with prostate cancer compared to controls. Serum vitamin C was significantly lower in patients with prostate cancer when compared to controls (p<0.01). Moreover, Se and Zn levels were also significantly lower, and levels of Ni, Co, and Cu were higher (p<0.001) in patients with prostate cancer than in controls. Fe levels were not significantly different in patients compared to controls (p>0.05). Our findings, together with the results of previous animal studies, suggest that the administration of vitamins A, C, and E, and Se and Zn may be beneficial in the prevention and treatment of human prostate cancer. PMID- 16475905 TI - Experimental and computer modelling speciation studies of the effect of pH and phosphate on the precipitation of calcium and magnesium salts in urine. AB - BACKGROUND: pH and phosphate concentration are the major determinants of precipitation in urine of the salts of calcium and magnesium. This study aims to model the process of salt precipitation and establish whether the acidification of urine samples is necessary for the accurate measurement of calcium and magnesium in a clinical laboratory setting. METHODS: Urine samples were collected from 21 patients, aliquots were taken from each patient sample and the pH was adjusted to cover the range 2.0-10.0. The analytical and biological variation for each analyte was established and used to calculate percentage changes and critical differences. The critical difference was used to assess whether there was a significant difference between acidified and un-acidified samples. The JESS (Joint Expert Speciation System) thermodynamic computer-modelling program was used to predict the distribution of salt species formed with varying pH values and phosphate levels in simulated urine. RESULTS: The results showed that at a pH greater than 6.5, measured calcium, magnesium and phosphate significantly decreased as a result of precipitation (p<0.0001), although the critical difference was generally not exceeded. Computer modelling showed that both pH and phosphate concentration affected the distribution of salt species formed, as well as the precipitation patterns of calcium and magnesium phosphates. Overall, calcium phosphate precipitation tends to predominate at lower phosphate concentrations and at pH values below about 6.5, while both calcium and magnesium phosphate precipitation occur at higher phosphate concentrations and pH values greater than 6.5. CONCLUSIONS: For accurate analysis of these analytes in urine, the pH should be routinely measured and acidification should be undertaken prior to analysis if the pH is greater than 6.5. Based on the findings of this study, acidification or the lack of it does not result in a clinically significant change in calcium, magnesium and phosphate measured in urine. This study also predicted the likely salt species formed at varying urinary pH values and phosphate concentrations. PMID- 16475906 TI - Total tau protein, phosphorylated tau (181p) protein, beta-amyloid(1-42), and beta-amyloid(1-40) in cerebrospinal fluid of patients with dementia with Lewy bodies. AB - The intra vitam diagnosis of different dementias is still based on clinical grounds. So far, no technical investigations have been available to support these diagnoses. For tau protein and beta-amyloid(1-42) in cerebrospinal fluid (CSF), promising results for the diagnosis of Alzheimer's disease (AD) have been reported; however, their differential diagnostic spectrum is limited, as was recently shown for dementia with Lewy bodies (DLB) and for AD. Therefore, further marker proteins have to be established to ameliorate, support, and differentiate these clinical diagnoses. We evaluated beta-amyloid(1-40) and phosphorylated tau protein (181p), in addition to total tau protein and beta-amyloid(1-42), in 20 patients with DLB, 34 AD patients, and 20 non-demented neurological controls (NDCs). All markers could differentiate between the dementia groups (AD, DLB) and the controls. AD and DLB could be differentiated only by levels of total tau protein and by the ratio total tau protein/phosphorylated tau protein. However, values still overlapped markedly. In some cases, tau protein levels in CSF may contribute to the clinical distinction between DLB and AD, but the value of the markers is still limited, especially because of mixed pathology. We conclude that more specific markers have to be established to differentiate between these diseases. PMID- 16475907 TI - Establishment of an indocyanine green test using an automatic chemistry analyzer. AB - The indocyanine green (ICG) clearance test has been used to assess the reserve of hepatic function. This method is based on the spectrometric measurement of its plasma concentration at maximum wavelength of 805 nm, which requires a spectrophotometer and associated maintenance. We established an ICG clearance test using a Toshiba 200FR automatic chemistry analyzer that can be tuned to a wavelength of approximately 805 nm. Five pooled sera spiked from 0 to 4.0 mg/dL were analyzed for linearity test and precision was determined at five levels in the range 0.1-2.0 mg/dL. The ICG retention rate at 15 min (R15) was determined for 38 patients using a conventional method and our method. The ICG clearance test using the automatic chemistry analyzer showed good linearity, and precision ranged from 0.3% to 1.0% for within-run CVs and from 0.6% to 4.7% for total CVs. The degree of agreement between the two methods was also acceptable (mean difference of 1.5%). It is expected that the ICG test using the automatic chemistry analyzer can replace the conventional ICG clearance test, considering the excellent agreement, good precision and linearity over a clinically relevant range. PMID- 16475908 TI - Relationship between serum sialic acids, sialic acid-rich inflammation-sensitive proteins and cell damage in patients with acute myocardial infarction. AB - The role of sialic acid (SA) in the pathogenesis of atherosclerosis and as a predictor of cardiovascular events has attracted much attention in recent years. However, most studies investigating the role of total and lipid-bound sialic acids (TSA and LSA) in the pathogenesis of atherosclerosis lack information on the reason for the elevated SA concentrations in coronary heart disease and myocardial infarction. Since the inflammation-sensitive proteins are glycoproteins with SA residues, an increase in their levels due to some type of acute-phase reaction or inflammation could be responsible for the elevated TSA levels in acute myocardial infarction (AMI). Elevated serum SA levels might also be due to either shedding or secretion of free SA from the cell or cell membrane surface if neuraminidase levels are increased, or to the release of cellular SA containing glycolipids and/or glycoproteins into plasma from myocardial cells after AMI. The aim of the present study was to investigate both the possible role of SA-rich inflammation-sensitive proteins and the cell damage due to elevated serum TSA levels in AMI. A possible role of serum LSA as an indicator of the shedding or secretion of SA from the cell or cell membrane surface in AMI was also evaluated. The study included 38 subjects with AMI and 32 healthy volunteers. Serum TSA and LSA were determined using the methods of Warren and Katopodis, respectively. The concentrations of serum SA-rich inflammation sensitive proteins, namely alpha1-antitrypsin, alpha2-macroglobulin and ceruloplasmin were determined immunoturbidimetrically. Our data showed that: a) mean levels of serum TSA and LSA and SA-rich inflammation-sensitive proteins in patients with AMI were significantly increased; and b) there was a significant positive correlation between TSA and LSA and alpha1-antitrypsin in patients with AMI. Since the transfer of free SA to lipoproteins is required for an increase in serum LSA levels, and free SA for this transfer can be provided by the secretion of SA from the cell, it is obvious that the shedding or secretion of SA from the cell membrane surface or release of cellular SA from cells into the bloodstream due to cell damage after AMI also occur after AMI. As a result, we can report that either the shedding or secretion of SA from the cell or cell membrane surface and the increased output of SA-rich inflammation-sensitive proteins may together be responsible for the elevated TSA levels in AMI. PMID- 16475909 TI - Ischemia-modified albumin and NT-prohormone-brain natriuretic peptide in peripheral arterial disease. AB - Cardiovascular disease is the leading cause of mortality and morbidity in Western countries. Despite its remarkable medical and social consequences, the prevalence of peripheral arterial disease (PAD) is often underestimated among atherosclerotic disorders. So far, little is known about the behavior of traditional and emerging markers of ischemic heart disease that should allow the reliable identification of PAD patients at increased risk of developing myocardial ischemia and heart failure or dysfunction. To investigate this topic, we measured cardiac troponin T (cTnT), ischemia-modified albumin (IMA) and NT prohormone-brain natriuretic peptide (NT-proBNP) in 35 consecutive patients with clinically ascertained PAD (stage 2-4, according to Leriche-Fontaine) asymptomatic for chest pain and current heart failure, and 20 controls displaying moderate to high cardiovascular risk factors (hypertension, diabetes, hyperlipidemia), but with no clinical evidence of PAD. Although the concentrations of cTnT and IMA were not statistically increased in PAD patients, NT-pro-BNP values were substantially higher in PAD patients than in controls (62.6 vs. 7.4 pmol/L, p<0.0001). The percentage of subjects displaying values exceeding the specific NT-proBNP diagnostic threshold (>14.8 pmol/L) was also significantly different between PAD patients and controls (74% vs. 10%, p<0.001). After excluding PAD patients exceeding the 0.01 ng/mL cTnT cutoff value indicative of current ischemic cardiac involvement, the median concentration of NT-proBNP remained statistically increased (28.0 vs. 5.8 pmol/L, p<0.0001). Taken together, these results indicate that NT-proBNP, but not IMA, is substantially increased in PAD patients. This finding suggests that such patients, even though asymptomatic, might develop myocardial dysfunction, and thus warrant further investigation. PMID- 16475910 TI - Nitration of human plasminogen by RAW 264.7 macrophages reduces streptokinase induced plasmin activity. AB - Despite many years of study, clinical trials of new drugs to prevent thrombosis have often been disappointing. Part of the problem lies in our incomplete understanding of the regulation of plasminogen activation and/or inhibition in vivo. We have previously shown that in vitro nitration of plasminogen in plasma by peroxynitrite resulted in decreased plasmin activity. We hypothesized that macrophages may be agents of plasminogen nitration and designed this study to prove this hypothesis. We first better characterized our previous observations using purified plasminogen instead of whole plasma, studied the time and concentration dependence of these reactions, and co-incubated plasminogen with macrophages, as well as with non-inflammatory cells as controls, to assess nitration and impaired activity. When plasminogen (10 micromol/L) is incubated in the presence of SIN-1 (0.01-2 mmol/L), plasmin activity (generated by streptokinase) is reduced in a time- and concentration-dependent fashion. We performed experiments incubating human plasminogen in the presence of murine RAW264.7 macrophages, allowing for free diffusion of reactive oxygen species, while preventing the action of proteases. In this way we show that incubation of plasminogen with macrophages also decreases plasmin activity, while increasing nitration of the molecule, an effect that is already apparent after 2 h and reaches a plateau of 60% inhibition after 24 h of incubation. This effect appears specific for macrophages, since 31EG4 murine mammary cells used in parallel and under the same conditions failed to produce any deleterious changes in plasminogen. Our data on quick functional inactivation of plasminogen by nitration, mediated by macrophages, adds a new pathophysiological dimension to our previous work showing plasminogen as a target for peroxynitrite damage. Nitrosative stress may be implicated in impaired fibrinolysis. New therapeutic approaches for nitrosative stress in atherosclerosis and diabetes should limit the formation of superoxides and peroxynitrite. PMID- 16475911 TI - Comparison of triglyceride concentration with lipemic index in disorders of triglyceride and glycerol metabolism. AB - BACKGROUND: In plasma, triglycerides (TG) are transported in lipoprotein particles (mainly chylomicrons, very low-density and low-density lipoprotein). Turbidimetry (bichromatically at 660 and 700 nm) allows measurement of the lipemic (L) index. We explored the use of this index, in combination with a TG assay, to detect errors due to non-fasting, to assess abnormalities in TG metabolism and to detect patients with glycerol kinase deficiency (GKD). METHODS: We collected 2441 patient samples. Normolipidemic (n=2347), type IV hyperlipidemic (n=80), postprandial samples (n=22) and serial dilutions of Intralipid with saline (n=6) were selected. One patient presenting with GKD was included, as well as two patients with type I and type V hyperlipoproteinemia, respectively. RESULTS: We introduced the use of the ratio between the logarithm of serum triglycerides and that of the L-index (TG/L ratio). CONCLUSION: Although the proposed TG/L-index ratio cannot be regarded as an alternative for the accurate diagnosis of lipid disorders, it provides additional information about TG-containing particles. PMID- 16475912 TI - Measurement of glycated hemoglobin (HbA1c) with an automated POCT instrument in comparison with HPLC and automated immunochemistry method: evaluation of the influence of hemoglobin variants. AB - Our study evaluates a fast and easy way to perform point-of-care testing (POCT) measurements of glycated hemoglobin HbA1c in comparison with an immunoassay on an automated biochemistry analyzer and cation exchange chromatography, the two methods routinely used in clinical laboratories for the measurement of HbA1c. A significant finding of our study is that although the POCT instrument insert claims that the method is not affected by the presence of HbS in the heterozygous state or in combination with beta-thalassemia, discrepant results were found in some cases with such hemoglobinopathies. In these cases, the two POCT and laboratory immunoassay methods showed clinically significant positive interferences with samples containing the HbS trait. We conclude that samples with the HbS trait should be interpreted with caution when tested using the POCT instrument. PMID- 16475913 TI - Analytical performance and clinical efficacy for cardiovascular risk estimation of an Olympus immunoturbidimetric high-sensitivity C-reactive protein assay. AB - Increased C-reactive protein (CRP) concentration within the reference interval (<10.0 mg/L) is a strong predictor of cardiovascular disease (CVD) in apparently healthy adults. Cutoff points for use of CRP in estimating CVD risk are <1, 1-3 and >3 mg/L for low, average and high relative risk, respectively. For measuring CRP concentrations to assess cardiovascular risk, high-sensitivity CRP (hsCRP) assays have been developed. The aim of this study was to evaluate the analytical performance and clinical efficacy for cardiovascular risk estimation of the Olympus immunoturbidimetric latex CRP assay (sensitive application). The comparative method used was the CardioPhase* hsCRP assay, approved by the Food and Drug Administration for use in CVD risk assessment. The imprecision of the Olympus hsCRP assay in the concentration range 0.2-10.0 mg/L was 0.38-8.16% within runs and 3.75-9.63% between runs. For method comparison studies, 194 fresh serum samples were selected to cover the interval 0.15-10.0 mg/L CRP. Comparison of the Dade Behring and Olympus methods was performed using weighted Deming regression analysis (slope 0.99 mg/L, intercept 0.002 mg/L, S(y,x)=0.02 mg/L, r=0.992) and a Bland-Altman relative difference plot (mean difference -0.002%, SD=0.040%). The agreement between the Dade Behring and Olympus methods for relative risk class assignments was 95.4%. Statistical analysis of the agreement between the two methods for each relative risk class showed that the differences between the methods were not statistically significant (p>0.10). Although previous reports found poor performance of the Olympus CRP tests for use in cardiovascular and peripheral vascular risk estimation, our study proved good analytical performance and clinical efficacy of the Olympus hsCRP assay for this use. PMID- 16475914 TI - Systemic matrix metalloproteinase-9 (MMP-9) levels as prognostic indexes of bone disease in patients with multiple myeloma. PMID- 16475916 TI - Society of Medical Biochemists of Serbia and Montenegro: 50-year anniversary. PMID- 16475917 TI - Nature's medicines: traditional knowledge and intellectual property management. Case studies from the National Institutes of Health (NIH), USA. AB - With the emergence and re-emergence of infectious diseases and development of multi-drug resistance, there is a dire need to find newer cures and to produce more drugs and vaccines in the pipeline. To meet these increasing demands biomedical researchers and pharmaceutical companies are combining advanced methods of drug discovery, such as combinatorial chemistry, high-throughput screening and genomics, with conventional approaches using natural products and traditional knowledge. However, such approaches require much international cooperation and understanding of international laws and conventions as well as local customs and traditions. This article reviews the forty years of cumulative experience at the National Institutes of Health (initiated by the National Cancer Institute) in natural products drug discovery. It presents (1) three major cooperative programs (2) the legal mechanisms for cooperation and (3) illustrative case studies from these programs. We hope that these discussions and our lessons learned would be helpful to others seeking to develop their own models of cooperation for the benefit of global health. PMID- 16475918 TI - Database mining for pKa prediction. AB - The acid dissociation constant (pKa) is the key parameter to define the extent of ionization of a drug molecule and is used for ADME properties evaluation via the pH-dependent distribution coefficient, logD. We present a method for pKa prediction using a predefined reference database and structural fingerprints based on a multilevel neighborhoods description of the ionizable atom(s). This database mining approach is suitable for screening large compound collections for HTS compound prioritization and external compound acquisition. In addition to pKa prediction it provides medicinal chemists rapid access to already available pKa measurements and hints for manipulating the chemical structure to increase or decrease pKa. PMID- 16475919 TI - Immobilisation of cardosin A in chitosan sponges as a novel implant for drug delivery. AB - Cardosin A is extracted from the pistils of the plant Cynara cardunculus L. and chitosan is a polysaccharide derived from chitin with valuable properties as a biomaterial. In this work we report our experiments on the synthesis of chitosan sponges and immobilisation of cardosin A, by entrapment. We observed that 10-15% of the incorporated cardosin A were released over 6 days of incubation. In addition, we could also note that this immobilisation procedure did not induce any specificity alterations on cardosin A. The specificity study of the enzyme, using beta-chain of oxidised insulin, showed that the immobilised and released enzymes have the same hydrolysis pattern as the free enzyme. The ability of this enzyme to hydrolyse type I collagen was maintained, after the immobilisation procedure. The biocompatibility in vivo of these sponges was evaluated by histological staining after implantation in rats submitted to abdominal surgery. Results of this study demonstrated that these chitosan sponges are very promising vehicles for the application of cardosin A, in abdominal cavity for prevention and reduction of the adhesions formation. PMID- 16475920 TI - Peptimmunology: immunogenic peptides and sequence redundancy. AB - Using short peptide fragments of proteins to elicit antibodies able to recognize the protein from which the peptide sequence was derived, is one of the main goals in immunotherapy today. Indeed, peptide-immunotherapy appears as an obliged way to obtain antibodies of predetermined specificity and exempt from the complications associated with whole cells/entire protein vaccines. However, effective peptide-immunotherapy remains an exciting theoretical speculation largely unrealized to date. The major obstacle in designing effective peptide vaccines is our incapacity to scientifically define peptide immunogenicity. This mini-review schematically describes: 1) the available methods to identify epitopic peptides; 2) the sequence redundancy concept as a possible basis for peptide immunogenicity. PMID- 16475921 TI - A computer-based approach to the rational discovery of new trichomonacidal drugs by atom-type linear indices. AB - Computational approaches are developed to design or rationally select, from structural databases, new lead trichomonacidal compounds. First, a data set of 111 compounds was split (design) into training and predicting series using hierarchical and partitional cluster analyses. Later, two discriminant functions were derived with the use of non-stochastic and stochastic atom-type linear indices. The obtained LDA (linear discrimination analysis)-based QSAR (quantitative structure-activity relationship) models, using non-stochastic and stochastic descriptors were able to classify correctly 95.56% (90.48%) and 91.11% (85.71%) of the compounds in training (test) sets, respectively. The result of predictions on the 10% full-out cross-validation test also evidenced the quality (robustness, stability and predictive power) of the obtained models. These models were orthogonalized using the Randic orthogonalization procedure. Afterwards, a simulation experiment of virtual screening was conducted to test the possibilities of the classification models developed here in detecting antitrichomonal chemicals of diverse chemical structures. In this sense, the 100.00% and 77.77% of the screened compounds were detected by the LDA-based QSAR models (Eq. 13 and Eq. 14, correspondingly) as trichomonacidal. Finally, new lead trichomonacidals were discovered by prediction of their antirichomonal activity with obtained models. The most of tested chemicals exhibit the predicted antitrichomonal effect in the performed ligand-based virtual screening, yielding an accuracy of the 90.48% (19/21). These results support a role for TOMOCOMD CARDD descriptors in the biosilico discovery of new compounds. PMID- 16475922 TI - Emerging Hsp90 inhibitors: from discovery to clinic. AB - Hsp90 is a chaperone with important roles in maintaining transformation and in elevating the survival and growth potential of cancer cells. Activation of signaling pathways mediated by Hsp90 protein clients is necessary for cell proliferation, regulation of cell cycle progression and apoptosis. Additionally, gain-of-function mutations responsible for transformation often require Hsp90 for the maintenance of their folded, functionally active conformations. These characteristics promise Hsp90 as an important target in cancer therapy and prompt for the identification, development and clinical translation of small molecule inhibitors of the chaperone. This review intends to update the reader on the status of several existing and emerging classes of direct inhibitors of Hsp90 ATPase activity. PMID- 16475923 TI - Peptide phage display: opportunities for development of personalized anti-cancer strategies. AB - Personalized medicine is critical for cancer patients, because (1) cancer is a highly heterogeneous disease with major molecular differences in the expression and distribution of tumor cell surface markers among patients with the same type and grade of cancer, (2) cellular mutations tend to accumulate as cancer progresses, further increasing tumor heterogeneity, and (3) currently used cancer therapies often are toxic to normal cells, causing severe side effects rarely seen in other diseases. To prevent side effects and to improve effectiveness, cytotoxic therapies should be targeted and each patient should be profiled for the presence of cancer targets before the therapy is administered. Phage display technology utilizes combinatorial libraries of proteins expressed on phage particles that can be selected for specific binding to cancer cells. Such cancer specific molecules can be used in a variety of applications, including identification of cell-specific targeting molecules; identification of cell surface biomarkers; profiling of specimens obtained from individual cancer patients, and the design of peptide-based anti-cancer therapeutics for personalized treatments. This review is focused on peptide phage display strategies that target cell surfaces because many biomarkers important in cancer are differentially expressed molecules located on the outside of the cell membranes. PMID- 16475924 TI - Gold coordination complexes as anticancer agents. AB - Metal ions are known to bind with nucleic acids and thereby alter their conformation and biological function. The metal ion-base interaction depends on the nature of both metal and bases; a certain site of coordination is preferred. One of the most notable successes for inorganic drugs has been the effectiveness of platinum complexes against cancer. These advances have spurred a surge of investigations to identify new inorganic agents for use in chemotherapy with improved specificity and decreased toxic side effects. Gold(I) and gold(III) complexes, the last isostructural and isoelectronic with platinum(II) complexes, are potentially attractive as anticancer agents. The design of an effective anticancer agent is a complicated game that must encompass not only the drug's inherent inhibitory properties but also its delivery, dosage, and residence time in vivo. Gold(I) and gold(III) complexes overcome some of these challenges by forming strong covalent attachments to targets. Au(III) isoelectronic with Pt(I1) d8 system usually forms square planar complexes in solution. Since the square planar geometry of Pt(II) is important for its action as an anticancer drug, Au(III) compounds also can be used for the same purpose with the added advantage of decreased toxicity. This, together with the recent finding that certain transitional metal complexes like Au and Pt complexes have been found to be potentially useful in cancer chemotherapy, created a renewed interest in the study of the interactions of metal ions with respect to the site of binding and the structure and stability of the complexes. This work was motivated by the thought that information on the variety of Au(III) complexes and their effects can be obtained by studying the properties of Au complexes with various ligands. Various studies in the past have shown that Au complexes are very attractive in view of their application as anticancer agents. PMID- 16475925 TI - Marine peptides and related compounds in clinical trial. AB - Advances in the sophisticated instruments for the isolation and characterization of marine natural products, and development in the biological assay systems, have resulted in the discovery of various compounds of biomedical application. Marine natural products have been a source of new leads for the treatment of many deadly diseases such as cancer, acquired immuno-deficiency syndrome (AIDS) etc. The compounds of marine origin are diverse in structural class from simple linear peptides to complex macrocyclic polyethers. Number of marine peptides have been isolated in recent years which exhibit potent biological activities, and many of the compounds showed promising anticancer activity. Didemnin was the first marine peptide that entered in human clinical trials in US for the treatment of cancer, and other anticancer peptides such as kahalalide F, hemiasterlin, dolastatins, cemadotin, soblidotin, didemnins and aplidine have entered in the clinical trials. Clinical status of anticancer marine derived peptides have been discussed and reviewed. PMID- 16475926 TI - Understanding apoptosis in terms of QSAR. AB - Apoptosis is a cellular mechanism by which unwanted, defective, or damaged cells are rapidly and selectively eliminated to maintain healthy tissue homeostasis in multi-cellular organisms. In this review, we have developed nineteen quantitative structure-activity relationships (QSAR) for different series of compounds with respect to their apoptotic activities against five different cancer cell lines that are T47D, ZR75-1, DLD-1, H1299, and HL60 in order to understand chemical biological interactions. PMID- 16475927 TI - Vitamin D analogs as anti-carcinogenic agents. AB - Deltanoids are the class of compounds comprising all natural and synthetic vitamin D molecules. The anti-proliferative, pro-differentiation, and pro apoptotic properties of deltanoids have garnered interest in the fields of cancer chemoprevention and chemotherapy. The naturally occurring, biologically active form of vitamin D, 1,25(OH)2D3, causes hypercalcemia at pharmacologically relevant doses which forms a major obstacle in the clinical development of this compound. Design of new deltanoids has shown promise in separating the beneficial effects from the toxic effects. The Vitamin D receptor (VDR) is a major target for deltanoid design, and the structural features of deltanoid binding have been described. Effective compounds must also exhibit beneficial pharmacokinetic properties in vivo, and the plasma vitamin D binding protein (DBP) is likely to play an important role in the success of deltanoids in the clinic. Further, dual strategies of avoiding vitamin D toxicity through altering the dosing schedule and using less toxic deltanoids are in development. The three main categories of structural modification to the vitamin D backbone include the C,D-ring, the A ring, and the C,D-ring side chain, and the ways each area has impacted efficacy and toxicity have been described through structure-activity relationships (SARs). Lastly, there is evidence that deltanoids can enhance the activity of other chemopreventive agents. The use of a cocktail approach will be discussed as a potential avenue for deltanoids in chemoprevention and chemotherapy. PMID- 16475928 TI - Towards selective Kir6.2/SUR1 potassium channel openers, medicinal chemistry and therapeutic perspectives. AB - ATP sensitive potassium (K(ATP)) channels have important functions in neuroendocrine tissue, in smooth and skeletal muscle and in the heart. In pancreatic beta cells the K(ATP) channels, which are formed by 4 ion channels (Kir6.2) and 4 regulatory sulfonylurea receptors (SUR1), control the glucose stimulated release of insulin. The Kir6.2/SUR1 K(ATP) channels are also present in the brain and in other neuroendocrine tissues. Blockers of Kir6.2/SUR1 channels, e.g. glibenclamide and repaglinide stimulate release of insulin and are used for treatment of type 2 diabetes. Openers of Kir6.2/SUR1 channels, e.g. diazoxide, have in contrast only found limited clinical use in treatment of hypersecretion of insulin associated with certain tumours (insulinoma) and genetic disorders (persistent hyperinsulinemia and hypoglycemia of infancy, PHHI). Recent studies have however, indicated that openers of Kir6.2/SUR1 channels could be useful in treatment of e.g. metabolic disorders and diseases of the CNS. The clinical use of diazoxide has been hampered by its lack of potency and selectivity giving rise to side effects, such as oedema and hirsutism and new selective openers of Kir6.2/SUR1 channels have been pursued. This has provided several structurally diverse series, which include 1,2,4-thiadiazine 1,1-dioxide derivatives, like BPDZ 62, BPDZ 73, NNC 55-0462, NNC 55-0118 and NN414, cyanoguanidines, nitropyrazoles and 4-sulfamoylphenylbenzamides. NN414 has been shown to be a potent and Kir6.2/SUR1 selective K(ATP) channels opener, which inhibits glucose stimulated insulin release in vitro and in vivo and which has beneficial effects on glucose homeostasis in preclinical and clinical studies. PMID- 16475930 TI - Dimeric and hybrid anti-Alzheimer drug candidates. AB - In the last decade much attention has been paid to the development of metabolically non-reversible dimeric or hybrid compounds, which combine two structural units of one or two lead compounds of interest for the treatment of Alzheimer's disease. As a consequence of their capability to simultaneously interact with two binding sites of the same biological target (the enzyme acetylcholinesterase in most cases), to expand their interaction in the main binding site of the target molecule, or to interact with two different biological targets of interest in the pathogenesis of the disease, these dimeric or hybrid compounds exhibit an improved pharmacological profile including high affinity interactions, additional non conventional actions or complementary actions, what makes them potential drug candidates for the treatment of Alzheimer's disease. Herein, we review from a structural point of view the main classes of dimeric or hybrid compounds developed for the treatment of Alzheimer's disease, along with the pharmacological profile of the most active compounds. PMID- 16475929 TI - Towards species-specific antifolates. AB - Dihydrofolate reductase (DHFR) plays an essential role in cellular biochemistry and has been a well-recognized drug target for over fifty years. Antifolate inhibitors of DHFR, including clinically used therapeutics such as methotrexate, trimethoprim, and pyrimethamine have been successful as anticancer, antibacterial, antifungal and antiparasitic agents. As resistant strains of these microorganisms evolve and as new disease threats arise, the need for new antifolates that are potent and specific for infectious organisms becomes more pressing. Several new antifolates have been reported over the past decade; many of these are potent against a particular species of DHFR, but achieving the goal of potency and selectivity has proven to be more difficult. This review will describe recent advances in attaining species selectivity in developing new antifolates. Specifically, advances in developing inhibitors against Pneumocystis jirovecii and Plasmodium falciparum, the causative agents in pneumocystis pneumonia and malaria, respectively, will be presented. PMID- 16475931 TI - Chemical Toxicity on HeLa Cells. AB - HeLa cells were named for Henrietta Lacks, who died in 1952 from an infection of a special type of cancer. Margaret Gey, her physician, started working with these cancer cells that are still used for medical research. In the present review, an attempt has been made to collect the data for the effects of different chemicals on HeLa cells and to discuss them by the formulation of a total number of 22 QSAR. PMID- 16475932 TI - Modulation of renal tubular cell survival: where is the evidence? AB - Tubular cell loss is prominent both in acute and chronic renal failure. Apoptosis and its regulatory mechanisms contribute to cell number regulation in the kidney. The potential role of apoptosis ranges from induction and progression to repair of renal injury. However, therapeutic interest has focused in preventing the apoptotic loss of tubular cells that leads to acute and chronic renal failure. Death ligands and receptors, such as tumor necrosis factor and Fas ligand, proapoptotic and antiapoptotic Bcl2 family members and caspases have all been shown to participate in apoptosis regulation in the course of renal cell injury. Nevertheless, the precise role of these proteins is unclear, and the participation of most known apoptosis regulatory proteins has not been studied. We now review the role of apoptosis in renal injury, the potential molecular targets of therapeutic intervention, the therapeutic weapons to modulate the activity of these targets and the few examples of therapeutic intervention on apoptosis, with emphasis in acute renal failure. PMID- 16475933 TI - Fluoroquinolones: an important class of antibiotics against tuberculosis. AB - Tuberculosis (TB) is a global health problem due to the lack of new drugs in the market and also due to the advent of multidrug resistant strains (MDR). This disease affects around 8 million people and kills almost 3 million people each year and it is estimated that there are 1 billion infected with TB worldwide. Due to this problem fluoroquinolones have attracted much attention as the new class of anti TB drugs due to their fewer toxic side effects, improved pharmacokinetic properties and extensive and potent activity against Gram-positive and Gram negative bacteria, including resistant strains. In this present review we report fluoroquinolones as a promising new class of anti TB. PMID- 16475934 TI - A role for leptin in the systemic inflammatory response syndrome (SIRS) and in immune response, an update. AB - Leptin was originally identified as an adipocyte-derived cytokine with a key role in the regulation of the energy balance. Subsequent research revealed that leptin's biological action is not restricted to its effects on appetite and food intake, but instead has a much more pleiotropic character. There is now ample evidence that leptin has important functions in reproduction, hematopoiesis, HPA axis endocrinology and angiogenesis. In this review we have focused on the effects of leptin in the antigen-specific immunity and in the inflammatory effector system. PMID- 16475935 TI - Agonists and antagonists of protease activated receptors (PARs). AB - Protease activated receptors (PARs) are a category of G-protein coupled receptors (GPCRs) implicated in the progression of a wide range of diseases, including thrombosis, inflammatory disorders, and proliferative diseases. Signal transduction via PARs proceeds via an unusual activation mechanism. Instead of being activated through direct interaction with an extracellular signal like most GPCRs, they are self-activated following cleavage of their extracellular N terminus by serine proteases to generate a new receptor N-terminus that acts as an intramolecular ligand by folding back onto itself and triggering receptor activation. Short synthetic peptides corresponding to this newly exposed N terminal tethered ligand can activate three of the four known PARs in the absence of proteases, and such PAR activating peptides (PAR-APs) have served as templates for agonist/antagonist development. In fact much of the evidence for involvement of PARs in diseases has relied upon use of PAR-APs, often of low potency and uncertain selectivity. This review summarizes current structures of PAR agonists and antagonists, the need for more selective and more potent PAR ligands that activate or antagonize this intriguing class of receptors, and outlines the background relevant to PAR activation, assay methods, and physiological properties anticipated for PAR ligands. PMID- 16475936 TI - Urotensin-II receptor antagonists. AB - Urotensin-II (U-II) is a "somatostatin-like" cyclic neuropeptide which was originally isolated from goby fish urophysis, and subsequently identified in other species, including man. The interest in human U-II (hU-II) has grown enormously in the last few years, following the identification of a specific human receptor (formerly identified as the GPR14/SENR orphan receptor), now referred to as UT receptor. The U-II/UT system seems to play an important role in cardiovascular functions. hU-II vasoconstrictive potency is reported to be an order of magnitude greater than that of endothelin-1 (ET-1), which would make it the most potent mammalian vasoconstrictor identified to date. hU-II also exerts potent inotropic effects in the human heart in vitro. On the basis of its spectrum of activities, hU-II has been suggested to modulate cardiovascular homeostasis and possibly to be involved in certain cardiovascular pathologies. Central nervous effects of U-II have also been described, in particular, intracerebroventricular administration promotes anxiogenic-like behaviors in rodents. Furthermore, UT receptor overexpression has been observed in some tumor cell lines. Therefore, specific and selective UT receptor antagonists provide useful tools for investigating the (patho)physiological role(s) of the U-II/UT receptor system. In this review we aim to provide an overview of the research in the area of UT receptor antagonists as well as the progress in understanding the role of the U-II/UT system in human (patho)physiology. PMID- 16475937 TI - Prediction oriented QSAR modelling of EGFR inhibition. AB - Epidermal Growth Factor Receptor (EGFR) is a high priority target in anticancer drug research. Thousands of very effective EGFR inhibitors have been developed in the last decade. The known inhibitors are originated from a very diverse chemical space but--without exception--all of them act at the Adenosine TriPhosphate (ATP) binding site of the enzyme. We have collected all of the diverse inhibitor structures and the relevant biological data obtained from comparable assays and built prediction oriented Quantitative Structure-Activity Relationship (QSAR) which models the ATP binding pocket's interactive surface from the ligand side. We describe a QSAR method with automatic Variable Subset Selection (VSS) by Genetic Algorithm (GA) and goodness-of-prediction driven QSAR model building, resulting an externally validated EGFR inhibitory model built from pIC50 values of a diverse structural set of 623 EGFR inhibitors. Repeated Trainings/Evaluations (RTE) were used to obtain model fitness values and the effectiveness of VSS is amplified by using predictive ability scores of descriptors. Numerous models were generated by different methods and viable models were collected. Then, intensive RTE were applied to identify ultimate models for external validations. Finally, suitable models were validated by statistical tests. Since we use calculated molecular descriptors in the modeling, these models are suitable for virtual screening for obtaining novel potential EGFR inhibitors. PMID- 16475938 TI - P2 receptors activated by uracil nucleotides--an update. AB - Pyrimidine nucleotides, including UTP, UDP and UDP-glucose, are important signaling molecules which activate G protein-coupled membrane receptors (GPCRs) of the P2Y family. Four distinct pyrimidine nucleotide-sensitive P2Y receptor subtypes have been cloned, P2Y2, P2Y4, P2Y6 and P2Y14. P2Y2 and P2Y4 receptors are activated by UTP (the P2Y2, and the rat but not the human P2Y4 receptor are also activated by ATP), the P2Y6 receptor is activated by UDP, and the P2Y14 receptor by UDP-glucose. Furthermore, non-P2Y GPCRs, the cysteinylleukotriene receptors (CysLT1R and CysLT2R) have been described to be activated by UDP in addition to activation by cysteinylleukotrienes. While P2Y2, P2Y4, and P2Y6 receptor activation results in stimulation of phospholipase C, the P2Y14 receptor is coupled to inhibition of adenylate cyclase. Derivatives and analogs of the physiological nucleotides UTP, UDP and ATP have been synthesized and evaluated in order to obtain enzymatically stable, subtype-selective agonists. The P2Y2 receptor agonists diuridine tetraphosphate (diquafosol) and the uracil-cytosine dinucleotide denufosol are currently undergoing clinical trials for dry eye disease, retinal detachment disease, upper respiratory tract symptoms, and cystic fibrosis, respectively. The first antagonists for P2Y2 and P2Y6 receptors that appear to be selective versus other P2Y receptor subtypes have recently been described. Selective antagonists for P2Y4 and P2Y14 receptors are still lacking. Uracil nucleotide-sensitive P2Y receptor subtypes may constitute future targets for the treatment of certain cancer types, vascular diseases, inflammatory diseases, and immunomodulatory intervention. They have also been proposed to play a role in neurodegenerative diseases. This article is an updated version of "P2 Pyrimidinergic Receptors and Their Ligands" by C. E. Muller published in Curr. Pharm. Des. 2002, 8, 2353-2369. PMID- 16475939 TI - HIV-1 reverse transcriptase: a therapeutical target in the spotlight. AB - Human Immunodeficiency Virus type 1 Reverse Transcriptase (HIV-1 RT) is one of the most important targets for treatment of Acquired Immune Deficiency Syndrome (AIDS). It catalyzes the reverse transcription of HIV-RNA into a double stranded DNA, and the knowledge of its substrate specificity and catalytic mechanism has guided the development of several inhibitors widely used on current HIV/AIDS therapy. However, mutations in HIV-1 RT structure can lead to the emergence of drug-resistant virus strains. The goal of this review is to summarize relevant structural features of HIV-1 RT and its inhibitors in such a way that this cost effective target in the development of new antiretroviral drugs is particularly highlighted. PMID- 16475940 TI - Taking advantage of viral immune evasion: virus-derived proteins represent novel biopharmaceuticals. AB - In healthy individuals, natural and adaptive immune responses are able to control virus entry into the host. In particular, CD8(+)-mediated cytotoxicity, sustained by the intervention of CD4+ cells, represents the major key event leading to virus eradication. On the other hand, viruses are able to evade from host immune response via several mechanisms, and special emphasis will be placed on hepatitis C virus and chronic Epstein-Barr infections also in view of personal data. Virokines, viroreceptors, and serpins greatly contribute to viral immune escape, and, among virokines, interleukin (IL)-10 has been object of intensive studies. Finally, all these products have been used as biopharmaceuticals, and, for instance, viral IL-10, chemokine-binding proteins, and serpins exhibit in animal models immunosuppressive, anti-inflammatory, and antiatherogenic activities. As far as their use in human trials is concernded, many cautions are required in order to avoid deleterious side effects and, in particular, the purity of the product, its route and frequency of administration, as well as the immune status of the patient should be taken into serious account. PMID- 16475941 TI - Progresses in the field of drug design to combat tropical protozoan parasitic diseases. AB - The progresses made in the field of drug design to combat tropical protozoan parasitic diseases, such as Chagas' disease, leishmaniasis, and sleeping sickness are discussed. This article is focused on different approaches based on unique aspects of parasites biochemistry and physiology, selecting the more promising molecular targets for drug design. In spite of the enormous amount of work on the above features, the chemotherapy for all of these diseases remains unsolved. It is based on old and fairly not specific drugs associated, in several cases, with long-term treatments and severe side effects. Drug resistance and different strains susceptibility are further drawbacks of the existing chemotherapy. In this review article, a thorough analysis of selected molecular targets, mainly those that are significantly different compared with the mammalian host or, even, are not present in mammals would be described in terms of their potencial usefulness for drug design. Therefore, this article covers rational approaches to the chemotherapeutic control of these parasitic infections, such as the progresses in the search for novel metabolic pathways in parasites that may be essential for parasites survival but with no counterpart in the host. Ergosterol biosynthesis is a very interesting example. There are many enzymes involved in this biosynthetic pathway such us squalene synthase, farnesylpyrophosphate synthase, and other enzymes that are able to deplete endogenous sterols will be treated in this article. The enzymes involved in trypanothione biosynthesis, glutathionyl spermidine synthetase and trypanothione synthetase do not have an equivalent in mammals, and therefore it can be predicted low toxicity for compounds that are able to produce highly selective inhibition. Trypanothione reductase (TR), glyceraldehyde-3-phosphate dehydrogenase, dihydrofolate reductase, prenyltransferases, ornithine decarboxylase, etc, will be thoroughly analyzed. The design of specific inhibitors of such metabolic activities as possible means of controlling the parasites without damaging the hosts will be presented. The recent advances in the biochemistry of pathogenic parasites including the discovery of novel organelles will be discussed. PMID- 16475942 TI - Herpesvirus saimiri-based gene delivery vectors. AB - Herpesviruses possess a number of characteristics which make them promising gene delivery vectors. These include their capacity to package large amounts of heterologous DNA and an ability to establish persistent, lifelong infections, where the viral genome remains as a circular non-integrated episome. Herpesvirus saimiri (HVS) is the prototype gamma-2 herpesvirus and is currently being developed as a potential gene delivery vector. In addition to the above properties, HVS-based vectors have the ability to infect a wide range of human cell lines and primary cultures with high efficiencies. Moreover, upon infection the viral genome persists as high copy number, circular, non-integrated episomes which segregate to progeny cells upon division. This allows the HVS-based vector to stably transduce a dividing cell population and provide sustained heterologous gene expression. As such, it offers the characteristics of an artificial chromosome combined with a highly efficient delivery system. This review aims to describe the assessment of HVS-based vectors in both in vitro and in vivo studies, highlighting new developments and possible applications for the treatment of genetic diseases. PMID- 16475943 TI - Targeting transcription factors for cancer gene therapy. AB - A high proportion of oncogenes and tumor suppressor genes encode transcription factors. Deregulated expression or activation and inactivation of transcription factors as well as mutations and translocations play critical roles in tumorigenesis. Furthermore, the majority of oncogenic signaling pathways converge on sets of transcription factors that ultimately control gene expression patterns resulting in tumor formation and progression as well as metastasis. Under normal physiological conditions whole sets of genes with similar functions are regulated by highly specific, tightly regulated upstream transcriptional regulators, whereas in cancer aberrant activation of these transcription factors leads to deregulated expression of multiple gene sets associated with tumor development and progression. The activity of these transcription factors can be modulated by multiple mechanisms including posttranslational modifications. Activation or inactivation of transcription factors promote cancer development, cell survival and proliferation and induce tumor angiogenesis. Since many of these transcription factors are inactive under normal physiological conditions and their expression and activities are tightly regulated, these transcription factors represent highly desirable and logical points of therapeutical interference in cancer development and progression. Three major families of transcription factors have emerged as important players in human cancer and are validated targets in drug discovery for cancer therapy: 1) the NF-kappaB and AP-1 families of transcription factors, 2) the STAT family members and 3) the steroids receptors. This review aims to elucidate the divergent molecular mechanisms involved in the deregulated activation of transcription factor signaling in malignant transformation, although additional transcription factor families such as the Ets factors, ATF family members, basic helix-loop-helix transcription factors etc. are additional critical transcriptional regulators in human cancer. We explore new approaches to specifically inhibit these transcription factors in cancer in order to validate them as a drug targets. Efforts to develop novel viral vectors for therapeutic applications are also discussed. PMID- 16475944 TI - Heparan sulphate proteoglycans and viral vectors : ally or foe? AB - The attachment of viruses to the host cell surface is a critical stage that will largely condition cell permissivity and productive infection. The understanding of such mechanisms is therefore essential for gene therapy applications involving viruses, as this step will influence both targeting and delivery efficiency of the gene of interest. Viral attachment depends upon the recognition and binding of viral envelope/capsid proteins to specific cellular receptors that can be from very diverse origins. Amongst them are heparan sulphate proteoglycans (HSPGs), a family of glycoproteins which, through the large binding properties of their heparan sulphate (HS) polysaccharide chains, serve as attachment receptor for a great number of viruses. The aim of this review is to provide an update on the multiple roles of HSPGs during viral infection, with a special focus on viruses used as gene delivery vectors. Consequences of HS binding for gene therapy applications will be assessed, as well as the various strategies that have been developed to potentiate the advantages or to overcome the drawbacks resulting from viral vector interaction with HS. PMID- 16475945 TI - Gene therapy strategies towards immune tolerance to treat the autoimmune diseases. AB - Autoimmune diseases such as type 1 diabetes and multiple sclerosis pose a significant health burden on our society. As a whole, autoimmune diseases affect approximately 6% of the population and are the third largest disease burden after heart disease and cancer. Such pathologic manifestations arise by way of damaging reactions of B-cell derived antibodies and/or T-cells to self-antigens and are triggered by genetic and environmental factors. Currently there is no known cure, with treatment restricted to toxic, long-term immunosuppressive regimes, replacement therapy and in intractable cases, transplantation of autologous or allogeneic haematopoietic stem cells. In experimental models of autoimmunity, gene therapeutic approaches have demonstrated promise in treating the autoimmune diseases. These include delivery of anti-inflammatory cytokines and exploitation of regulatory T cells. However, none of these approaches provide lasting, long term benefit. We hypothesise that therapeutically transduced haematopoietic stem cells followed by transplantation is an alternative strategy to establish permanent immune tolerance that can not only prevent autoimmunity but also cure these diseases. Our approach is focused on directing autoimmune disease-specific autoantigen expression in the thymus by genetic manipulation of haematopoietic stem cells to establish molecular chimeras. Our hypothesis originates from experimental studies with a mouse model of experimental autoimmune gastritis (EAG) and more recently with the non-obese diabetic (NOD) mouse model for type 1 diabetes (T1D). PMID- 16475946 TI - Synthetic and natural polycations for gene therapy: state of the art and new perspectives. AB - Currently, the major drawback of gene therapy is the gene transfection rate. The two main types of vectors that are used in gene therapy are based on viral or non viral gene delivery systems. There are several non-viral systems that can be used to transfer foreign genetic material into the human body. In order to do so, the DNA to be transferred must escape the processes that affect the disposition of macromolecules. These processes include the interaction with blood components, vascular endothelial cells and uptake by the reticuloendothelial system. Furthermore, the degradation of therapeutic DNA by serum nucleases is also a potential obstacle for functional delivery to the target cell. Cationic polymers have a great potential for DNA complexation and may be useful as non-viral vectors for gene therapy applications. The objective of this review was to address the state of the art in gene therapy using synthetic and natural polycations and the latest strategies to improve the efficiency of gene transfer into the cell. PMID- 16475947 TI - MDA-7/IL-24-based cancer gene therapy: translation from the laboratory to the clinic. AB - Despite recent advances in treatment strategies, the overall 5-year survival rate for patients with common epithelial cancers is poor largely because of the difficulty in treating metastatic cancers. Therefore, therapeutic agents are urgently needed that can effectively inhibit both primary epithelial tumors and their metastases. One such agent that has shown promise in preclinical studies is the tumor suppressor/cytokine, melanoma differentiation associated gene-7 also known as interleukin-24 (mda-7/IL-24). Preclinical studies from our and other laboratories have shown that overexpression of MDA-7/IL-24 causes a strong tumor- suppressive effect in many human cancer cells but spares normal cells. This gene therapy also enhances the tumor-suppressive activity of radiotherapy and chemotherapy. Secreted MDA-7 protein that is glycosylated also has been shown to have potent antiangiogenic activity both in vitro and in vivo. Studies examining the immune properties of mda-7 have shown that MDA-7/IL-24 unlike the related IL 10, functions as a Th1 cytokine. Recently, an MDA-7 protein-mediated "bystander effect" on tumor cells has been documented. Building on these findings we successfully completed a Phase I clinical trial of adenovirus-based mda-7 cancer therapy that confirmed the safety of this gene therapy. Phase II trials evaluating the efficacy of mda-7-based gene therapy are warranted. The outcome of such ongoing mda-7-based gene therapy trials will allow us to better understand this therapy's clinical utility. PMID- 16475948 TI - Plasmid-mediated muscle-targeted gene therapy for circulating therapeutic protein replacement: a tale of the tortoise and the hare? AB - There is now conclusive evidence that gene therapy can lead to real clinical benefit. Initial enthusiasm has been muted by set-backs related to viral vectors including retroviral oncogenesis and adenoviral inflammatory response. Plasmid mediated muscle-targeted gene transfer offers the potential of a cost-effective pharmaceutical grade therapy delivered by simple intramuscular injection without the need for anaesthetic, cell culture, transplantation or immunosuppression. This approach is particularly appropriate for long-term circulating therapeutic protein replacement currently requiring repeated injection therapy. Wide-ranging clinical applications include haemophilia, chronic anaemia, growth hormone deficiency and diabetes. Inadequate transgene expression, unregulated protein delivery and immune response have been major limiting factors. Recent innovations including in situ electroporation enabling sustained systemic protein delivery within the therapeutic range are reviewed. Pharmacological and physiological approaches to regulation are discussed in addition to the role of innate and humoral immunity. Translation of advances in all of these areas to clinical success will enable muscle-targeted gene therapy to capitalise on its inherent strengths and realise its long-standing promise. PMID- 16475949 TI - Targeting DNA repair proteins: a promising avenue for cancer gene therapy. AB - Enhanced DNA repair in many cancer cells can be correlated to the resistance to cancer treatment, and thus contributes to a poor prognosis. Ionizing radiation and many anti-cancer drugs induce DNA double-strand breaks (DSBs), which are usually regarded as the most toxic types of DNA damages. Repair of DNA DSBs is vital for maintaining genomic stability and hence crucial for survival and propagation of all cellular organisms. Therefore, reducing the capacity of cancer cells to repair DSBs could sensitize tumors to radio/chemotherapy. Many investigators have used gene therapy strategies to down-regulate or inactivate proteins involved in the repair of DSBs in order to reduce the survival of cancer cells. Herein, are reviewed several protein candidates that have been targeted by different gene therapy approaches. Results obtained from in vitro and in vivo experiments are presented and discussed in the perspective of potential gene therapy clinical trials. PMID- 16475950 TI - The neuroendocrine system as a model to evaluate experimental gene therapy. AB - The implementation of experimental gene therapy in animal models of neurological diseases is an area of growing interest. Although the neuroendocrine system offers unique advantages for the assessment of in vivo gene therapy, little work has been done in this model. Here we review the core of documented studies in which in vivo gene therapy has been implemented in the neuroendocrine system of rodent models. In the hypothalamus, restorative gene therapy has been successfully implemented in Brattleboro rats, an arginine vasopressin (AVP) mutant which suffers from diabetes insipidus, in Koletsky (fa(k)/fa(k)) and in Zucker (fa/fa) rats which have leptin receptor mutations that render them obese, hyperphagic and hyperinsulinemic. In the above models, viral vectors expressing AVP, leptin receptor b and proopiomelanocortin, respectively were stereotaxically injected in the relevant hypothalamic regions. In rats, aging brings about a progressive degeneration and loss of hypothalamic tuberoinfundibular dopaminergic neurons, which are involved in the tonic inhibitory control of prolactin secretion and lactotrophic cell proliferation. Stereotaxic injection of an adenoviral vector expressing Insulin-like Growth Factor-I (IGF-I) was able to correct their chronic hyperprolactinemia and restore tuberoinfundibular dopaminergic (TIDA) neuron numbers. In young and old F-344 male rats, Glial Cell Line-derived Neurotrophic Factor (GDNF) gene delivery in the hypothalamus induced body weight loss. These results suggest that further implementation of gene therapy strategies in neuroendocrine models may be highly rewarding. PMID- 16475951 TI - RNA based gene therapy for dominantly inherited diseases. AB - There are numerous examples in the literature of gene therapy applications for recessive disorders. There are precious few instances, however, of studies conducted to treat dominantly inherited pathologies. The reasons are simple: there are fewer cases of dominantly inherited diseases on one hand, but mostly it is far easier to correct recessive mutations than dominant ones. Typically recessive mutations cause a loss of (or reduced) gene function which can be compensated for by introduction of a replacement allele into the cell. In contrast, dominant negative mutations not only display impaired function, but also exhibit a novel one that is pathologic to the cell. Treating these conditions by gene therapy implies silencing the dominant allele without altering the expression of the wild-type gene. We describe here different strategies aimed at silencing dominant mutations through mRNA destruction and provide examples of their application to known autosomal dominant diseases. An overview of the most common molecular tools (antisense DNA and RNA, ribozymes and RNA interference) suitable to utilize these strategies is also presented and we discuss the relevant aspects involved in the choice of a particular approach in a gene therapy experiment. PMID- 16475952 TI - Intracompartmental delivery of CNTF as therapy for Huntington's disease and retinitis pigmentosa. AB - Ciliary neurotrophic factor (CNTF) is a cytokine with neurotrophic activity across a broad spectrum of peripheral and central nervous system (CNS) cells. While its therapeutic potential for CNS diseases has been clear for sometime, the blood brain barrier (BBB) hinders the systemic delivery of CNTF and direct bolus injections are not suitable due to the short half-life of CNTF. One means of overcoming the BBB while providing continuous delivery of CNTF is with immunoisolated cellular implants that produce and deliver CNTF directly to the region of interest. Cells can be protected from host rejection by encapsulating, or surrounding, them within an immunoisolatory, semipermeable membrane that admits oxygen and required nutrients and releases bioactive cell secretions, but restricts passage of larger cytotoxic agents from the host immune defense system. The selective membrane eliminates the need for chronic immunosuppression of the host and allows the implanted cells to be obtained from nonhuman sources. In this review we discuss cell immunoisolation for Huntington's disease and retinitis pigmentosa. These two indications are highlighted because of extensive pre clinical data supporting the general concept and recent clinical data that both strengthens the theoretical role of CNTF for treating neurodegeneration and justifies additional clinical evaluation in these and other diseases. PMID- 16475954 TI - Recent strategies for potentiation and facilitation of antidepressant treatment: introduction. PMID- 16475955 TI - Pindolol augmentation of antidepressant response. AB - Pindolol, a partial beta-adrenoceptor/5-HT1A receptor antagonist was first used to accelerate the onset of action of antidepressant drugs in 1994. Since then, it has been used in more than a dozen controlled trials to examine whether it can reduce the lag to clinical improvement, and/or improve the clinical response in treatment-resistant patients. A recent metaanalysis concluded that pindolol accelerates the antidepressant response but does not increase the effectiveness of SSRIs in unresponsive patients. Several studies have examined the pharmacology of pindolol to clarify the neurobiological basis of its clinical action. Pindolol was initially used due to its ability to block 5-HT1A receptor-mediated responses and to enhance the neurochemical effects of SSRIs. In transfected cells, however, pindolol is a weak (20-25%) partial agonist at 5-HT1A receptors and, as such, its actions greatly depend on the system used. In line with this, other reports have also shown that pindolol can reduce serotonergic cell firing when given alone. Positron emission tomography (PET) scan studies have shown that pindolol displays a preferential occupancy of pre- vs. postsynaptic 5-HT1A receptors, although the overall occupancy is lower than desirable, which suggests that higher doses (e.g., 15 mg/day) may be more effective than the currently used 7.5 mg daily dosage. However, given the complex pharmacology of pindolol, it is hoped that new developments in this field can proceed through the use of a) selective and silent 5-HT1A receptor antagonists in combination with SSRIs, or b) dual action agents (SSRI+5-HT1A receptor blockers). PMID- 16475956 TI - Catecholaminergic strategies for the treatment of major depression. AB - Although the selective serotonin reuptake inhibitors have become the first line medications for the treatment of depression, drugs primarily targeting the norepinephrine (NE) and/or the dopamine catecholaminergic systems are also effective. These include selective NE reuptake inhibitors, such as desipramine and reboxetine, the NE releaser bupropion and the alpha2-adrenergic antagonists mianserin and mirtazapine. Dopamine type 2 agonists are also effective in treating depression, although they are rarely used. Since the NE, dopamine and serotonin systems have reciprocal interactions, it is virtually impossible to act on a specific neuronal element without affecting in a cascade effect the two other systems. In this review, the primary actions of the catecholaminergic strategies upon their acute and long-term administration are described, as well as their impact on other systems. Their use in treatment-resistant depressed patients is also addressed. PMID- 16475957 TI - Potentiation of antidepressant-like activity with lithium: mechanism involved. AB - In the last decade, many augmentation strategies have been developed to increase the activity of antidepressant drugs or to reduce their long onset of action by acting on different targets. One of the first augmentation strategy used in psychiatric disorders is coadministration of lithium and antidepressant drugs. However, the underlaying mechanism of action involved in the potentiatory effect of lithium is still unclear and many hypotheses have been suggested such as activity on BDNF, ACTH, thyroid hormones and serotonin neurotransmission. All these systems being embedded in each other, we focused on the 5-HT neurotransmission-increase induced by lithium treatment. Based on neurobiochemical and behavioral results we tried to better understand its mechanism of action and we concluded that effect of lithium on 5-HT neurotransmission could be linked to a partial agonist activity on 5-HT1B autoreceptors, or to a modulatory activity on these receptors, located in the cortical area in the case of a short term treatment, or in the hippocampus in the case of a long term treatment. We also suggested that the anti-manic effect of lithium was linked to this activity on 5-HT1B receptors, occurring this time on 5 HT1B postsynaptic (heteroreceptors on dopaminergic pathways) receptors levels. PMID- 16475958 TI - Modulation of dopamine transmission by 5HT2C and 5HT3 receptors: a role in the antidepressant response. AB - Dopaminergic mesolimbic and mesocortical systems are fundamental in hedonia and motivation. Therefore their regulation should be central in understanding depression treatment. This review highlights the dopaminergic activity in relation to depressive behavior and suggests two putative receptors as potential targets for research and development of future antidepressants. In this article we review data that describe the role of serotonin in regulating dopamine release, via 5HT2C and 5HT3 receptors. This action of serotonin appears to be linked to depressive-like behavior and to onset of behavioral effects of antidepressants in an animal model of depression. We suggest that drugs or strategies that decrease 5HT2C and increase 5HT3 receptor-mediated dopamine release in the limbic areas of the brain may provide a fast onset of therapeutic effect. Clinical and basic research data supporting this hypothesis are discussed. PMID- 16475959 TI - Serotonin-dopamine interaction as a focus of novel antidepressant drugs. AB - Central serotonergic and dopaminergic systems play a critical role in the regulation of normal and abnormal behavior. Recent evidence suggests that a dysfunction of dopamine (DA) and serotonin (5-HT) neurotransmitter systems contributes to various pathological conditions. Among the multiple classes of 5 HT receptors described in the central nervous system, much attention has been devoted to the role of 5-HT2 receptor family in the control of central dopaminergic activity, because of the moderate to dense localization of both transcript and protein for 5-HT2A and 5-HT2C receptors in the substantia nigra (SN) and ventral tegmental area (VTA), as well as their terminal regions. Moreover, modulation of 5-HT2 receptor function by various drugs that has been shown to influence DA function in these brain areas is thought to be important in motor activation, motivation, and reward. Indeed, a number of electrophysiological and biochemical data have shown that 5-HT2C receptor agonists decrease, while 5-HT2C receptor antagonists enhance mesocorticolimbic DA function. Recent studies have focused on the functional interaction between the serotonergic and dopaminergic systems to explain the mechanism of the antidepressant action of SSRIs and 5-HT2 antagonists. In this article, the most relevant data regarding the role of these receptors in the control of brain DA function are reviewed, and the importance of this subject in the search of new antidepressant drugs is discussed. PMID- 16475960 TI - Microdialysis approach to study serotonin outflow in mice following selective serotonin reuptake inhibitors and substance P (neurokinin 1) receptor antagonist administration: a review. AB - Classical antidepressant drugs such as Selective Serotonin Reuptake Inhibitors (SSRIs) display several disadvantages, e.g., the onset of action (2 to 3 weeks) to start clinical benefits is too long, and a significant proportion of patients do not respond to this monotherapy. Several strategies have been proposed to overcome these problems, notably the use of potentiating agents, which combined with SSRIs, augment or accelerate their established antidepressant activity. Recent clinical trials proposed that compounds with dual action on both central serotonin (5-HT) and noradrenaline (NA) systems would have a faster action than SSRIs alone. Preclinical electrophysiological and neurochemical studies demonstrated that the putative new class of antidepressants, substance P (neurokinin 1) NK1 receptor antagonists, enhance brain monoaminergic neurotransmissions by reducing the sensitivity of 5-HT1A autoreceptors in the Dorsal Raphe Nucleus, and possibly alpha2 autoreceptors in the Locus Coeruleus. However, in several clinical studies, a similar delay of therapeutic effects has been reported with NK1 receptor antagonists and SSRIs. Recently intracerebral in vivo microdialysis studies were performed to examine the effects of genetic or pharmacological blockade of Substance P (SP)/ NK1 neurotransmission on SSRIs induced increases in extracellular 5-HT levels in awake, freely moving mice. New evidences suggest that the combination of a NK1 receptor antagonist with a SSRI should benefit to depressed patients. This review describes our current knowledge of the role of SP and its preferred NK1 receptors mainly in the modulation of brain serotonergic activity. PMID- 16475962 TI - Preventing type 2 diabetes in high risk patients: an overview of lifestyle and pharmacological measures. AB - BACKGROUND: Type 2 diabetes mellitus (T2DM) is a common disease that is associated with an increased risk of vascular complications. The incidence of T2DM is also increasing. It follows that T2DM prevention is important. METHODS: Relevant articles (review articles, randomised studies and large cohort and case control studies) were identified through a Medline search (up to March 2005). RESULTS: The first trials on T2DM prevention were based on lifestyle intervention. The results of these studies were impressive since they demonstrated that even a small reduction in weight could significantly reduce the incidence of T2DM. However, the main disadvantage of lifestyle measures is that they are difficult to achieve and sustain. Therefore, pharmacological interventions have also been evaluated. The results of trials using metformin, orlistat, nateglinide, acarbose, thiazolidinediones, hormone replacement therapy, statins or fibrates are either encouraging or require more extensive evaluation. In addition, studies using antihypertensive drugs (mainly angiotensin-converting enzyme inhibitors and angiotensin II receptor antagonists) showed that these drugs could also reduce the progression to T2DM in high risk individuals. CONCLUSIONS: T2DM has major quality of life and cost implications. Therefore, more research is needed to establish safe and cost effective ways to prevent this modern epidemic. PMID- 16475961 TI - Basic mechanisms of augmentation of antidepressant effects with thyroid hormone. AB - The thyroid hormone triiodothyronine (T3) has been used both to augment and accelerate the clinical effects of antidepressants, particularly the tricyclics. More recent work indicates that it may have similar actions with regard to the SSRIs. Two main mechanisms have been put forward to explain its antidepressant actions, (a) an action at the nuclear level involving stimulation of gene transcription, (b) an action at the cell membrane level involving potentiation of neurotransmission. In particular, there is considerable evidence for potentiation by T3 of the actions of the neurotransmitter 5-HT or serotonin. This evidence, which is mainly based on in vivo microdialysis studies, is reviewed, and evidence based on human and animal neuroendocrine studies considered. The effects of T3, alone and together with the SSRI fluoxetine, on mRNA levels for the 5-HT1A and 5 HT1B autoreceptors, which mediate serotonergic neurotransmission by feedback actions at the levels of cell firing(somatodendritic 5-HT1A autoreceptors) and neurotransmitter release (nerve terminal 5-HT1B autoreceptors) were also determined. Administration of a combination of fluoxetine and T3 induced reductions in the transcription of these autoreceptors, which may explain the clinical potentiating effects of this combination, and thus link the nuclear and neurotransmitter hypotheses of T3 action. PMID- 16475964 TI - Efficient medicinal chemistry. PMID- 16475963 TI - Enhancing the site-specific targeting of macromolecular anticancer drug delivery systems. AB - The application of macromolecules as vehicles for anticancer drug delivery is a burgeoning field of interest. One of the hallmarks of using such systems, however, is that they must be capable of site-specific drug delivery. As such, augmenting the targeting of drug delivery systems to specified sites is paramount. To date, a number of synthetic strategies have been utilized to introduce targeting moieties to macromolecular drug delivery systems to enhance specific targeting. This scheme frequently involves the introduction of some type of biologically recognizable marker to the delivery system. Biological evaluations have substantiated the rationale that introducing targeting groups can significantly increase specificity. This concise review will attempt to encompass what strategies have been done to increase the specificity of macromolecular anticancer drug delivery systems along with their biological activities. PMID- 16475965 TI - The integration of process R&D in drug discovery--challenges and opportunities. AB - In today's situation where a lot of attention is put on the whereabouts of the pharmaceutical industry, especially focusing on productivity, pricing policies, time lines, and competition, there is an increased need for a critical revision of work practices in the business. The prevailing prioritization of time-to market is now more and more shifting over to also put quality, risk management, and effectiveness/efficiency in the limelight. Resources in terms of people and money will continue to be constrained and, therefore, best collaborative principles have to be adopted between different parts of the organization. Only by operating this way will we maximize the output. One of the most important key performance indicators in pharma R&D is the number of newly appointed candidate drugs (CDs). However, it is not only a matter of counting numbers but, more so, to nominate compounds with the best properties and likelihood to survive. In that vein the demands on Process R&D have gone up considerably over recent years and there is now a pronounced need to make forecasts on cost of goods for the API (active pharmaceutical ingredient), scalability issues, IP matters, route design etc. On top of this, there is as always an expectation that the supply of material needed to conduct the various studies is timely, fully reliable, and flexible, even if volumes and delivery dates fluctuate widely. To successfully be able to cope with this challenging and sometimes stressful situation a back integration into earlier parts of Drug Discovery is a must and, hence, connecting to new projects will have to be initiated already during the LO-stage (lead optimization). The consequences of this and its further implications will constitute the core part of the paper. PMID- 16475966 TI - Achieving maximum ROI from corporate databases: exploiting your databases with integrated querying for better decision-making. AB - In order to increase the rate of drug discovery, pharmaceutical and biotechnology companies spend billions of dollars a year assembling research databases. Current trends still indicate a falling rate in the discovery of New Molecular Entities (NMEs). It is widely accepted that the data need to be integrated in order for it to add value. The degree to which this must be achieved is often misunderstood. The true goal of data integration must be to provide accessible knowledge. If knowledge cannot be gained from these data, then it will invalidate the business case for gathering it. Current data integration solutions focus on the initial task of integrating the actual data and to some extent, also address the need to allow users to access integrated information. Typically the search tools that are provided are either restrictive forms or free text based. While useful, neither of these solutions is suitable for providing full coverage of large numbers of integrated structured data sources. One solution to this accessibility problem is to present the integrated data in a collated manner that allows users to browse and explore it and also perform complex ad-hoc searches on it within a scientific context and without the need for advanced Information Technology (IT) skills. Additionally, the solution should be maintainable by 'in-house' administrators rather than requiring expensive consultancy. This paper examines the background to this problem, investigates the requirements for effective exploitation of corporate data and presents a novel effective solution. PMID- 16475967 TI - Application and utilization of chemoinformatics tools in lead generation and optimization. AB - The process of Drug Discovery is a complex and high risk endeavor that requires focused attention on experimental hypotheses, the application of diverse sets of technologies and data to facilitate high quality decision-making. All is aimed at enhancing the quality of the chemical development candidate(s) through clinical evaluation and into the market. In support of the lead generation and optimization phases of this endeavor, high throughput technologies such as combinatorial/high throughput synthesis and high throughput and ultra-high throughput screening, have allowed the rapid analysis and generation of large number of compounds and data. Today, for every analog synthesized 100 or more data points can be collected and captured in various centralized databases. The analysis of thousands of compounds can very quickly become a daunting task. In this article we present the process we have developed for both analyzing and prioritizing large sets of data starting from diversity and focused uHTS in support of lead generation and secondary screens supporting lead optimization. We will describe how we use informatics and computational chemistry to focus our efforts on asking relevant questions about the desired attributes of a specific library, and subsequently in guiding the generation of more information-rich sets of analogs in support of both processes. PMID- 16475968 TI - Improving synthetic efficiency using the computational prediction of biological activity. AB - A process has been developed whereby libraries of compounds for lead optimization can be synthesized and screened with greater efficiency using computational tools. In this method, analogues of a lead chemical structure are considered in the form of a virtual library. Less than 1/3 of the library is selected as a training set by clustering the compounds and choosing the centroid of each cluster. This training set is then used to generate a model using PLS regression upon the experimental values from that assay using 1D/2D descriptors. The model is applied to the remaining compounds (the test set) for which assay values are predicted and a rank ordering established. An example of this was a set of 169 PDE4 inhibitors. A predictive model was achieved using a training set of 52 compounds. When applied to the remaining 117 compounds this model allowed a rank ordering of these compounds for synthesis and testing. Selecting the top 33 compounds of the test set gives 78% of the compounds with the desired activity (hits) by synthesizing only 50% of the library, including the training set. Selecting the top 59 of the test set gives 97% of the hits from only 67% of the library. This process succeeds by avoiding two principal weaknesses of 2D descriptors: lack of interpretation and lack of extrapolation. Two principal assumptions of QSAR are shown to be unnecessary; removing descriptor redundancy does not improve fit and a predictive r2 greater than 0.5 is not necessary if rank-ordering is desired. PMID- 16475969 TI - Comparison of methods for sequential screening of large compound sets. AB - Sequential screening is an iterative procedure that can greatly increase hit rates over random screening or non-iterative procedures. We studied the effects of three factors on enrichment rates: the method used to rank compounds, the molecular descriptor set and the selection of initial training set. The primary factor influencing recovery rates was the method of selecting the initial training set. Rates for recovering active compounds were substantially lower with the diverse training sets than they were with training sets selected by other methods. Because structure-activity information is incrementally enhanced in intermediate training sets, sequential screening provides significant improvement in the average rate of recovery of active compounds when compared with non iterative selection procedures. PMID- 16475970 TI - A collaborative hit-to-lead investigation leveraging medicinal chemistry expertise with high throughput library design, synthesis and purification capabilities. AB - High throughput screening (HTS) campaigns, where laboratory automation is used to expose biological targets to large numbers of materials from corporate compound collections, have become commonplace within the lead generation phase of pharmaceutical discovery. Advances in genomics and related fields have afforded a wealth of targets such that screening facilities at larger organizations routinely execute over 100 hit-finding campaigns per year. Often, 10(5) or 10(6) molecules will be tested within a campaign/cycle to locate a large number of actives requiring follow-up investigation. Due to resource constraints at every organization, traditional chemistry methods for validating hits and developing structure activity relationships (SAR) become untenable when challenged with hundreds of hits in multiple chemical families per target. To compound the issue, comparison and prioritization of hits versus multiple screens, or physical chemical property criteria, is made more complex by the informatics issues associated with handling large data sets. This article describes a collaborative research project designed to simultaneously leverage the medicinal chemistry and drug development expertise of the Novartis Institutes for Biomedical Research Inc. (NIBRI) and ArQule Inc.'s high throughput library design, synthesis and purification capabilities. The work processes developed by the team to efficiently design, prepare, purify, assess and prioritize multiple chemical classes that were identified during high throughput screening, cheminformatics and molecular modeling activities will be detailed. PMID- 16475971 TI - Interactive tools for risk reduction and efficiency improvements in medicinal chemistry. AB - There are many decisions and risks associated with the design and development of new pharmaceutical agents. To help improve decision-making, and reduce the associated risks--prior to synthesis, we have developed interactive web-browser tools for: (i) tracking, searching, clustering and categorizing (by reactive moieties) chemical reactants, (ii) interactively assessing risks, either synthetic--based on prior experience, absorption following oral administration- based on rules of 5, or diversity, and (iii) a complete architecture for enumerating, analyzing, submitting and plating large combinatorial or small biased libraries. We believe the implementation of this highly interactive system has given our scientists a competitive advantage by maintaining their focus on the lowest risk, highest quality molecules throughout the research process. PMID- 16475972 TI - Functional characterisation of homomeric ionotropic glutamate receptors GluR1 GluR6 in a fluorescence-based high throughput screening assay. AB - We have constructed stable HEK293 cell lines expressing the rat ionotropic glutamate receptor subtypes GluR1(i), GluR2Q(i), GluR3(i), GluR4(i), GluR5Q and GluR6Q and characterised the pharmacological profiles of the six homomeric receptors in a fluorescence-based high throughput screening assay using Fluo-4/AM as a fluorescent Ca2+ indicator. In this assay, the pharmacological properties of nine standard GluR ligands correlated nicely with those previously observed in electrophysiology studies of GluRs expressed in Xenopus oocytes or mammalian cells. The potencies and efficacies displayed by the agonists (S)-glutamate, (S) quisqualate, kainate, (RS)-AMPA, (RS)-ATPA, (RS)-ACPA] and (S)-4-AHCP at the six GluRs were in concordance with electrophysiological studies. Furthermore, the Ki values exhibited by the competitive antagonists NBQX and (RS)-ATPO were also in agreement with findings of previous studies. Finally, the effects of various concentrations of Ca2+ in the assay buffer and of the allosteric modulators cyclothiazide and concanavalin A on GluR signalling were examined. This study represents the most elaborate functional characterisation of multiple AMPA and KA receptor subtypes in the same assay reported to date. We propose that high throughput screening of compound libraries at the six GluR-HEK293 cell lines could be helpful in the search for structurally and pharmacologically novel ligands acting at the receptors. PMID- 16475973 TI - Rho GTPases: promising cellular targets for novel anticancer drugs. AB - Ras-homologous (Rho) GTPases play a pivotal role in the regulation of numerous cellular functions associated with malignant transformation and metastasis. Rho GTPases are localized at membranes and become activated upon stimulation of cell surface receptors. In their GTP-bound (=active) state, Rho proteins bind to effector proteins, thereby triggering specific cellular responses. Members of the Rho family of small GTPases are key regulators of actin reorganization, cell motility, cell-cell and cell-extracellular matrix (ECM) adhesion as well as of cell cycle progression, gene expression and apoptosis. Each of these functions is of importance for the development and progression of cancer. Furthermore, Rho guanine exchange factors (GEFs) are often oncogenic and the expression level of Rho GTPases frequently increases with malignancy. Rho proteins also affect cellular susceptibility to DNA damaging agents, including antineoplastic drugs and ionizing radiation (IR). Thus, modulation of Rho driven mechanisms may influence the therapeutic efficiency and/or the side effects of conventional antineoplastic therapy. Because of their pleiotropic functions, Rho proteins appear to be promising targets for the development of novel anticancer drugs. Experimental approaches to inhibit Rho (and Ras) have focused on the attenuation of their C-terminal isoprenylation. This is because C-terminal lipid modification is required for correct intracellular localization and function of Rho/Ras. Inhibitors of farnesyltransferase (FTI), geranylgeranyltransferase (GGTI) as well as of HMG-CoA-reductase (i. e. statins) have been investigated with respect to their usefulness in tumor therapy. The studies showed that these compounds affect tumor progression and furthermore have impact on the frequency of cell death induced by tumor therapeutics. A possible drawback of inhibitors of isoprenylation is their poor selectivity for individual Rho GTPases. Therefore, specific inhibitors of individual Rho functions (notably RhoA-, RhoB-, Rac1- or Cdc42-related functions) are predicted to be of great therapeutic benefit. Indeed, compounds developed as specific inhibitors of the RhoA-effector molecule Rho-kinase (ROK) have been demonstrated to exert anti-metastatic activity in vivo. PMID- 16475974 TI - Anti-cancer therapy: targeting the mevalonate pathway. AB - The mevalonate pathway has become an important target for anti-cancer therapy. Manipulation of this pathway results in alteration of malignant cell growth and survival in cell culture and animal models, with promising potential for application in human cancers. Mevalonate is synthesized from 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA). Mevalonate is further metabolized to farnesyl pyrophosphate (FPP), which is the precursor for sterols. In addition, the farnesyl moiety from FPP is utilized for post-translational modification of proteins including small GTPases, such as Ras and Ras related proteins, which play a role in malignant transformation of cells. FPP is a precursor for geranylgeranyl pyrophosphate (GGPP), which is similarly involved in post translational modification of proteins. There has been intense interest in manipulating the pathway through HMG-CoA reductase inhibition. More recently, the focus has been on manipulating the pathway by post-translational modification of key regulatory proteins through farnesyl prenyl transferase (FPTase) or geranylgeranyl prenyl transferase (GGPTase) inhibition. This review focuses on the mevalonate pathway and the application of rational drug therapies to manipulate this pathway. Included in the review are a summary of agents demonstrating success in preclinical investigations such as; farnesyl transferase inhibitors, geranylgeranyl transferase inhibitors, dual inhibitors, statins, bisphosphonates, histone deacetylase inhibitors and other compounds. While these agents have shown preclinical success, translation to success in clinical trials has been more difficult. These clinical trials are reviewed along with evaluation of some of the potential problems with these agents in their clinical application. PMID- 16475975 TI - Estrogen receptor alpha: impact of ligands on intracellular shuttling and turnover rate in breast cancer cells. AB - Estrogen receptors (alpha and beta) are members of the steroid/thyroid nuclear receptors superfamily of ligand-dependent transcription factors. Impact of the alpha isoform of estrogen receptor (ER) on breast cancer etiology and progression is now well established. Current therapeutic strategy to treat ER-positive breast cancer relies on the blockade of ER trancriptional activity by antiestrogens. Data accumulated during the last five years on the mechanism of action of ER enable one to foresee new strategies. These data indeed reveal that ER is not statically bound to DNA at promoter sites of genes regulating cell proliferation and/or differentiation, but rather behaves as a very mobile protein continuously shuttling between targets located within various cellular compartments (i.e. membrane, microsomes, nucleus...). This allows the receptor to generate both non genomic and genomic responses. Ligands, growth factors and second messengers produced downstream of activated membrane receptors modulate ER-mediated responses by interfering with the traffic patterns of the receptor, as well as by locally blocking its transient anchorage. Changes in ER turnover rate associated with these regulatory processes seem also to strongly influence the ability of the receptor to mediate gene transactivation. The present paper surveys these biological data and analyzes how they may be integrated into new drug design programs aimed at expanding our therapeutic armamentarium against breast cancer. PMID- 16475976 TI - The stem cell factor receptor/c-Kit as a drug target in cancer. AB - Tyrosine phosphorylation has a key role in intracellular signaling. Inappropriate proliferation and survival cues in tumor cells often occur as a consequence of unregulated tyrosine kinase activity. Much of the current development of anti cancer therapies tries to target causative proteins in a specific manner to minimize side-effects. One attractive group of target proteins is the kinases. c Kit is a receptor tyrosine kinase that normally controls the function of primitive hematopoietic cells, melanocytes and germ cells. It has become clear that uncontrolled activity of c-Kit contributes to formation of an array of human tumors. The unregulated activity of c-Kit may be due to overexpression, autocrine loops or mutational activation. This makes c-Kit an excellent target for cancer therapies in these tumors. In this review we will highlight the current knowledge on the signal transduction molecules and pathways activated by c-Kit under normal conditions and in cancer cells, and the role of aberrant c-Kit signaling in cancer progression. Recent advances in the development of specific inhibitors interfering with these signal transduction pathways will be discussed. PMID- 16475977 TI - Disruption of metabolic pathways--perspectives for the treatment of cancer. AB - Several growth-promoting signaling pathways have tight molecular connections with metabolic-related signal transduction systems. By controlling these pathways, cancer cells gain autonomy over energy-acquiring systems and, thus, expand their potential for proliferation. Here, we discuss the use of drug and antisense oligonucleotide approaches to inhibit metabolic pathways in cancer cells and their potential use in the therapeutics of cancer. PMID- 16475978 TI - Comparison of JNK (c-Jun N-terminal kinase)-like MAPK (mitogen-activated protein kinase) phosphorylation between immobilized cultures and Couette-type shear reactor cultures of Taxus cuspidata (Japanese yew) cells. AB - The signal mechanism of Taxus cuspidata (Japanese yew) cells involved in response to shear stress and immobilized matrices was investigated. Western-blot analysis showed that the immobilized cultures altered the degree of phosphorylation of the JNK (c-Jun N-terminal kinase)-like and a 41 kDa p38-like MAPKs (mitogen-activated protein kinases), particularly the 52 and 45 kDa JNK-like MAPKs. The increased rotation speeds up-regulated the degree of phosphorylation of 45 and 47 kDa JNK like MAPKs, whereas the 41 kDa p38-like MAPK was not significantly changed. The level of phosphorylation of JNK-like MAPKs in the outer zone cells of immobilized matrices was the highest among the different zone cells, which was identical with that of T. cuspidata cells exposed to hydrodynamic shear stresses. The highly specific p38- or JNK-MAPK inhibitors strongly reduced respectively the p38- or JNK-like MAPK phosphorylation, which also demonstrated that there were no cross reactions between the p38-like MAPK and JNK-like MAPK in T. cuspidata cells. By a comparison of the effects of the immobilization and laminar shear stress on the phosphorylation of p38- and JNK-like MAPKs in T. cuspidata cells, these findings suggested that the JNK-like MAPK signal pathways may be involved in T. cuspidata cell response to the hydrodynamic shear stress, rather than to the p38-like MAPK. PMID- 16475979 TI - Dipeptidyl peptidases 8 and 9: specificity and molecular characterization compared with dipeptidyl peptidase IV. AB - Dipeptidyl peptidases 8 and 9 have been identified as gene members of the S9b family of dipeptidyl peptidases. In the present paper, we report the characterization of recombinant dipeptidyl peptidases 8 and 9 using the baculovirus expression system. We have found that only the full-length variants of the two proteins can be expressed as active peptidases, which are 882 and 892 amino acids in length for dipeptidyl peptidase 8 and 9 respectively. We show further that the purified proteins are active dimers and that they show similar Michaelis-Menten kinetics and substrate specificity. Both cleave the peptide hormones glucagon-like peptide-1, glucagon-like peptide-2, neuropeptide Y and peptide YY with marked kinetic differences compared with dipeptidyl peptidase IV. Inhibition of dipeptidyl peptidases IV, 8 and 9 using the well-known dipeptidyl peptidase IV inhibitor valine pyrrolidide resulted in similar K(i) values, indicating that this inhibitor is non-selective for any of the three dipeptidyl peptidases. PMID- 16475980 TI - Participation of a galactose-specific C-type lectin in Drosophila immunity. AB - A galactose-specific C-type lectin has been purified from a pupal extract of Drosophila melanogaster. This lectin gene, named DL1 (Drosophila lectin 1), is part of a gene cluster with the other two galactose-specific C-type lectin genes, named DL2 (Drosophila lectin 2) and DL3 (Drosophila lectin 3). These three genes are expressed differentially in fruit fly, but show similar haemagglutinating activities. The present study characterized the biochemical and biological properties of the DL1 protein. The recombinant DL1 protein bound to Escherichia coli and Erwinia chrysanthemi, but not to other Gram-negative or any other kinds of microbial strains that have been investigated. In addition, DL1 agglutinated E. coli and markedly intensified the association of a Drosophila haemocytes derived cell line with E. coli. For in vivo genetic analysis of the lectin genes, we also established a null-mutant Drosophila. The induction of inducible antibacterial peptide genes was not impaired in the DL1 mutant, suggesting that the galactose-specific C-type lectin does not participate in the induction of antibacterial peptides, but possibly participates in the immune response via the haemocyte-mediated mechanism. PMID- 16475981 TI - Modelling insecticide-binding sites in the voltage-gated sodium channel. AB - A homology model of the housefly voltage-gated sodium channel was developed to predict the location of binding sites for the insecticides fenvalerate, a synthetic pyrethroid, and DDT an early generation organochlorine. The model successfully addresses the state-dependent affinity of pyrethroid insecticides, their mechanism of action and the role of mutations in the channel that are known to confer insecticide resistance. The sodium channel was modelled in an open conformation with the insecticide-binding site located in a hydrophobic cavity delimited by the domain II S4-S5 linker and the IIS5 and IIIS6 helices. The binding cavity is predicted to be accessible to the lipid bilayer and therefore to lipid-soluble insecticides. The binding of insecticides and the consequent formation of binding contacts across different channel elements could stabilize the channel when in an open state, which is consistent with the prolonged sodium tail currents induced by pyrethroids and DDT. In the closed state, the predicted alternative positioning of the domain II S4-S5 linker would result in disruption of pyrethroid-binding contacts, consistent with the observation that pyrethroids have their highest affinity for the open channel. The model also predicts a key role for the IIS5 and IIIS6 helices in insecticide binding. Some of the residues on the helices that form the putative binding contacts are not conserved between arthropod and non-arthropod species, which is consistent with their contribution to insecticide species selectivity. Additional binding contacts on the II S4-S5 linker can explain the higher potency of pyrethroid insecticides compared with DDT. PMID- 16475982 TI - Phosphatidylinositol synthesis is essential in bloodstream form Trypanosoma brucei. AB - PI (phosphatidylinositol) is a ubiquitous eukaryotic phospholipid which serves as a precursor for messenger molecules and GPI (glycosylphosphatidylinositol) anchors. PI is synthesized either de novo or by head group exchange by a PIS (PI synthase). The synthesis of GPI anchors has previously been validated both genetically and chemically as a drug target in Trypanosoma brucei, the causative parasite of African sleeping sickness. However, nothing is known about the synthesis of PI in this organism. Database mining revealed a putative TbPIS gene in the T. brucei genome and by recombinant expression and characterization it was shown to encode a catalytically active PIS, with a high specificity for myo inositol. Immunofluorescence revealed that in T. brucei, PIS is found in both the endoplasmic reticulum and Golgi. We created a conditional double knockout of TbPIS in the bloodstream form of T. brucei, which when grown under non-permissive conditions, clearly showed that TbPIS is an essential gene. In vivo labelling of these conditional double knockout cells confirmed this result, showing a decrease in the amount of PI formed by the cells when grown under non-permissive conditions. Furthermore, quantitative and qualitative analysis by GLC-MS and ESI MS/MS (electrospray ionization MS/MS) respectively showed a significant decrease (70%) in cellular PI, which appears to affect all major PI species equally. A consequence of this fall in PI level is a knock-on reduction in GPI biosynthesis which is essential for the parasite's survival. The results presented here show that PI synthesis is essential for bloodstream form T. brucei, and to our knowledge this is the first report of the dependence on PI synthesis of a protozoan parasite by genetic validation. PMID- 16475983 TI - Self-reported weight perceptions, dieting behavior, and breakfast eating among high school adolescents. AB - This study explored the relationships among weight perceptions, dieting behavior, and breakfast eating in 4597 public high school adolescents using the Centers for Disease Control and Prevention Youth Risk Behavior Survey. Adjusted multiple logistic regression models were constructed separately for race and gender groups via SUDAAN (Survey Data Analysis). Adjusted odds ratios [ORs] and 95% confidence intervals were calculated to determine the strength of relationships. Approximately 42% of the sample reported not eating breakfast within the past 5 days, while 41% were trying to lose weight, and 37% were dieting to lose weight. Excessive dietary practices (eg, fasting, taking diet pills or laxatives, and vomiting to lose weight) were reported by approximately 25% of the sample. When compared to those eating breakfast within the past 5 days, all race and gender groups that did not report eating breakfast were significantly more likely to report fasting to lose weight (ORs = 1.70-2.97). In addition, all race/gender groups, with the exception of black females, were significantly more likely to perceive themselves as overweight (ORs = 1.44-1.61) and trying to lose weight (ORs = 1.40-1.72). Among males, not eating breakfast was significantly associated with taking diet pills to lose weight (ORs = 2.31-2.40), eating fewer calories to lose weight (ORs = 1.38-1.49), and inversely associated with trying to gain weight (ORs = 0.71-0.74). Results suggest that these adolescents may be skipping breakfast as part of a patterned lifestyle of unhealthy weight management and that efforts to encourage youth to eat breakfast will likely not ameliorate all dietary challenges that appear beyond the scope of increased breakfast offerings. PMID- 16475984 TI - The politics of sexual orientation issues in American schools. AB - Schools are increasingly expected to address the needs of gay, lesbian, bisexual, and transgender students. However, the controversial nature of sexual orientation programs and policies makes this a politically sensitive undertaking. This empirical study analyzes the extent to which public school districts across the United States have implemented policy recommendations and describes, according to 4 theoretical policy models, factors that influence their ability to do so. The survey found that most districts have not institutionalized recommended policies or programs. Recommendations for school health professionals based on factors found to be significantly associated with the implementation of programs are discussed. PMID- 16475985 TI - Follow-up comparisons of intervention and comparison schools in a state tobacco prevention and control initiative. AB - This study provides information about prevention and control practices in intervention and comparison secondary schools 2 years after the start-up of the Texas Tobacco Prevention Initiative. The intervention, which was funded through the Texas Department of State Health Services, consisted of guidance, training, technical assistance, and reimbursement of approximately 2000 dollars per year for program expenses. Self-administered written surveys for Principals and Health Coordinators, based on the School Health Education Profile Tobacco Module, were designed for periodic assessment of the status of school programs. Surveys were sent in 2002 to intervention (n = 74) and comparison (n = 60) schools. Response to the Principal Survey was received from 109 (81%) schools, and response to the Health Coordinator Survey was received from 84 (63%) schools. Survey analysis showed that intervention schools more frequently (p or=95th percentile) was 24%, more than twice that of the national prevalence of 10% for 2- to 5-year olds documented by NHANES (1999-2002) and 3 times that of the 1998-1999 ECLS prevalence estimate of 8% for 5- to 7-year olds in the Midwest region. The data reported here document that nearly one quarter of children entering school in Chicago are already overweight. This clearly establishes a need for local schools to develop protocols and procedures to support the physical and mental health needs of affected and at-risk children. The findings also make it plain that ongoing weight status monitoring is needed and that current plans to implement this should go forward. PMID- 16475987 TI - Moderating effects of gender on alcohol use: implications for condom use at first intercourse. AB - This study examined whether the effects of level of alcohol consumption on condom use at first sex depend on adolescents' gender, utilizing data from Wave I of the National Longitudinal Study of Adolescent Health (Add Health). Compared to girls who did not consume any alcohol, inebriated girls were significantly less likely to use a condom at first intercourse (odds ratio = 0.41, p 70 mmHg with high-volume fluid therapy and dobutamine may help to reduce the severity of myopathy. PMID- 16476003 TI - A comparison of extradural and intravenous methadone on intraoperative isoflurane and postoperative analgesia requirements in dogs. AB - OBJECTIVE: To compare the effects of intravenous (IV) and extradural (ED) methadone on end-tidal isoflurane concentration (Fe'ISO) and postoperative analgesic requirements in dogs undergoing femoro-tibial joint surgery. STUDY DESIGN: Randomized, blinded, clinical study. Animals Twenty-four healthy client owned dogs undergoing surgical repair of ruptured cruciate ligaments. METHODS: Dogs were randomly assigned to two groups of 12 animals and received either ED or IV methadone (0.3 mg kg(-1) diluted with saline to 0.2 mL kg(-1)). Pre anaesthetic medication was IV acepromazine (0.05 mg kg(-1)). Anaesthesia was induced with propofol and maintained initially with an Fe'ISO of 1.0% delivered in oxygen. Methadone was injected with the dogs in sternal recumbency; the observer was unaware of the administration route. At 10 minutes (stimulation 1) and 20 minutes (stimulation 2) after methadone administration pelvic limb reflexes were tested by digit-clamping. The time at skin incision (stimulation 3), joint-capsule incision (stimulation 4), tibial tuberosity drilling (stimulation 5), fabellar suturing (stimulation 6) and extracapsular tightening (stimulation 7) were noted. Changes in heart rate (HR) and respiratory rate and arterial blood pressure associated with surgery were recorded along with the corresponding Fe'ISO. After 20 minutes of anaesthesia, Fe'ISO was decreased to the minimum required to maintain stable anaesthesia. Immediately after tracheal extubation, 1, 2, 3 and 6 hours postoperatively and on the morning after surgery, the degree of pain present was assessed using a numerical rating scale. The HR, respiratory rates and blood pressure were also recorded at these times. Serum cortisol and blood glucose concentrations were measured before pre-anaesthetic medication and at each postoperative pain scoring interval except at 1 and 2 hours. Ketoprofen (2 mg kg(-1)), carprofen (4 mg kg(-1)) or meloxicam (0.2 mg kg( 1)) were given by subcutaneous injection whenever pain scoring indicated moderate discomfort was present. RESULTS: Controlled ventilation was required in six dogs which stopped breathing after IV methadone. The median Fe'ISO at stimulus 5 was 1.0% in the IV and 0.83% in the ED group. At stimulus 6, Fe'ISO was 1.0% in the IV and 0.8% in the ED group; the difference was statistically significant (p or =10,000 Joule/m(2)) ultraviolet-B (UVB) irradiated allogeneic leukocytes in rodent models have been shown to induce immunologic tolerance that is mediated by allospecific regulatory CD4(+) T cells. Whether these regulatory T cells recognize alloantigens through the direct or indirect pathway of allorecognition is controversial. Here, we demonstrate that the proliferative response obtained in standard primary mixed leukocyte reactions (MLRs) with human peripheral blood mononuclear cells (PBMCs) reflected a CD4(+) T cell-dependent direct pathway of allorecognition and that high-dose UVB irradiation of PBMCs totally inhibited their capacity to induce a proliferative alloresponse. Re-stimulation with gamma-irradiated PBMCs from the same allogeneic donor (secondary MLR) elicited a proliferative and Th1-deviated response that was similar to the response induced in unprimed PBMCs. Finally, high-dose UVB was found to induce a rapid and massive apoptosis of irradiated PBMCs. Collectively, these data indicate that leukocytes irradiated with high-dose UVB are unable to prime for unresponsiveness or immune deviation in T cells directly recognizing allogeneic major histocompatibility complex molecules. Because it is well established that antigens within transfused apoptotic cells are captured by resident tolerogenic spleen dendritic cells, we propose that tolerance induced by transfusions of high-dose UVB-irradiated leukocytes primarily involve T cells indirectly recognizing alloantigens. PMID- 16476008 TI - The nontoxic CTA1-DD adjuvant enhances protective immunity against Helicobacter pylori infection following mucosal immunization. AB - Safe and efficacious adjuvants are much needed to facilitate the development of mucosal vaccines. Here, we have asked whether our nontoxic vaccine adjuvant, CTA1 DD, can enhance protective immunity against Helicobacter pylori infection. Intranasal immunizations with H. pylori lysate together with CTA1-DD-adjuvant induced significant protection in C57Bl/6 mice, almost as strong as similar immunizations using cholera toxin (CT)-adjuvant. Protection remained strong even at 8 weeks postchallenge and the bacterial colonization was reduced by 20-fold compared to lysate-immunized controls. Although CTA1-DD was designed to bind to B cells, microMT mice developed significant, but lower, level of protection following immunization. Intranasal immunizations with CT adjuvant in C57Bl/6 mice resulted in the development of severe postimmunization gastritis at 2 and 8 weeks postchallenge, whereas the degree of gastritis was substantially lower in the CTA1-DD-immunized mice. Protection induced by both CTA1-DD- and CT adjuvant was associated with a strong local infiltration of CD4(+) T cells in the gastric mucosa, and recall responses to specific Ag elicited substantial IFN-gamma production, indicating Th1-dominance. These findings clearly demonstrate that CTA1-DD adjuvant is a promising candidate to be further exploited in the development of a mucosal vaccine against H. pylori infection. PMID- 16476010 TI - Elimination of CD4+ CD25+ regulatory T cells breaks down reovirus type 2 triggered and CpG ODN-induced prolonged mild autoimmune insulitis in DBA/1 mice. AB - We have reported previously that subclinical prolonged mild T helper (Th) 1 dependent autoimmune insulitis with impaired glucose tolerance in wealing DBA/1J mice, which is induced by the combined effects of reovirus type 2 (Reo-2) and synthetic 20-base oligodeoxynucleotides with CpG motifs (CpG ODN) (control mice). Compared with the control mice, newborn mice treated with monoclonal antibody (MoAb) against mouse CD25(+) CD4(+) T cells together with Reo-2 and CpG ODN greatly reduced the absolute number of splenic CD25(+) T cells and resulted in the development of severe insulitis, leading to an overt early diabetes. Moreover, the treatment of the MoAb increased production of interferon-gamma (IFN gamma) and decreased that of interleukin-4 (IL-4) and transforming growth factor beta1 (TGF-beta1) and developed high titre of autoantibodies against pancreatic islet cells. These evidences suggest that CD4(+) CD25(+) T cell may, at least in part, maintain tolerance to Reo-2-triggered and CpG ODN-induced prolonged mild Th1-dependent autoimmune insulitis, leading to the overt disease. This system may give a novel model to elucidate the mechanisms of the development of overt diabetes from borderline subclinical diabetes in virus-triggered autoimmune type I diabetes in human. PMID- 16476009 TI - Serotonin transporter mRNA expression is decreased by lamivudine and ribavirin and increased by interferon in immune cells. AB - Clinical reports document that depression as a side effect is more prevalent in hepatic patients given interferon (IFN)-alpha therapy than in those given lamivudine. The mechanisms, however, are poorly understood. Serotonin transporter (5-HTT), via uptake of serotonin (5-HT) into presynaptic serotoninergic neurons, is an initial action site for antidepressants. Real-time polymerase chain reaction (PCR) was used to quantify 5-HTT mRNA expression in immune cells in order to evaluate whether 5-HTT acted as an indicator of depression. Results showed that the 5-HTT mRNA expression was much higher in T-cell and B-cell lines than that in a monocytic cell line. Treatment with either lamivudine or ribavirin reduced the 5-HTT mRNA expression, protein level and 5-HT uptake in T-cell line. Treatment with IFN-alpha, however, increased those levels in the same group. A similar effect was observed in peripheral blood mononuclear cells (PBMC). Mimicking clinical use by treating PBMC with a combination of IFN-alpha and ribavirin increased the 5-HTT mRNA expression level. Our study indicates that these therapeutic drugs regulate 5-HTT expression, which implies that 5-HTT might be a trait marker in IFN-alpha-induced depression after hepatic therapy. PMID- 16476011 TI - Hydrogen peroxide increases interleukin-12 p40/p70 molecular ratio and induces Th2-predominant responses in mice. AB - To investigate the influence of oxidative stress on the immune response, mice were injected with H(2)O(2), and peritoneal macrophages were isolated and stimulated in vitro with lipopolysaccharide (LPS). H(2)O(2) significantly augmented both interleukin (IL)-12p40 and IL-12p70 production and increased the p40/p70 molecular ratio. This was confirmed by mRNA analysis, which showed that H(2)O(2) increased LPS-induced mRNA expression of both IL-12p40 and IL-12p35 subunits with an increased p40/p35 ratio. Analysis of anti-ovalbumin (OVA) antibodies revealed that H(2)O(2) injection significantly increased the production of type 2 helper T cell (Th2)-associated antibody classes [immunoglobulin (Ig)E and IgG1] but not a Th1-associated antibody class (IgG2a). To confirm the Th2-predominant immune response, we analyzed the profile of cytokine production by spleen T cells of OVA-immunized and H(2)O(2)-injected mice. H(2)O(2) significantly increased the production of IL-4 but not that of interferon-gamma. Together, these results suggest that H(2)O(2)-induced overproduction of IL-12p40 promotes the Th2-predominant response through increased production of IL-12p40-homodimers, which could serve as an antagonist of the Th1-inducing cytokine IL-12p70. PMID- 16476012 TI - Mannan-binding lectin in children with chronic gastritis. AB - The involvement of mannan-binding lectin (MBL) insufficiency in the pathogenesis of chronic gastritis (CG) in children was investigated. Blood samples were collected from 78 paediatric patients suffering from CG associated with Helicobacter pylori infection (group Hp(+)) and from 41 with the disease not associated with such an infection (group Hp(-)). Control group consisted of 77 children. The frequency of mbl-2 gene mutations and serum protein concentrations did not differ significantly in both groups as compared with controls. An expression of mbl-2 gene in gastric biopsies of CG patients was demonstrated. It was found to be stronger in H. pylori-infected children. The results presented in this paper suggest that MBL deficit/dysfunction probably does not contribute to an increased risk of CG (both associated and not associated with H. pylori infection) in children. However, MBL opsonic effect and/or the lectin pathway of complement activation may be taken into account as possible host defence mechanisms in gastric patients. PMID- 16476013 TI - Interferon-gamma gene (T874A and G2109A) polymorphisms are associated with microscopy-positive tuberculosis. AB - Genetic susceptibility to tuberculosis includes several unknown yet different loci each contributing to a small extent. Intronic polymorphisms within the interferon-gamma (IFN-gamma) gene IFNG T+874A and IFNG G+2109A correlate with the IFN-gamma production in vitro, and the frequency of potential high IFN-gamma producers was previously reported by others to be lower in patients than in controls from Sicily. The aim of this study was to determine whether there is an association between polymorphisms in the IFN-gamma gene and predisposition to tuberculosis. We analysed two IFNG SNPs (T+874A and G+2109A) in patients (n = 253) hospitalized in Rijeka (Croatia) and controls (n = 519) from the same area. One-fifth of the controls were healthy contacts of the diseased, and the rest were blood donors. IFNG alleles, their predicted haplotypes or genotypes were not associated with disease susceptibility. Thus, we could not reproduce results from Sicilian case-control study. However, T/T+874 (possible high IFN-gamma producer) and +874A/A (putative low producer) genotypes were associated with microscopically positive-negative forms of disease. Haplotypes (T+874A and G+2109A) based on a prediction by software phase and subsequent genotype analysis corroborated these findings. Patients had significantly higher frequency of genotypes without T at +874 (AA/AA; AA/AG and AG/AG) in microscopy- or bacterial culture-positive groups compared with their negative counterparts. These data suggest an association with disease severity rather than susceptibility to tuberculosis in Croatian Caucasian population. PMID- 16476014 TI - Interferon-gamma receptor-1 gene promoter polymorphisms (G-611A; T-56C) and susceptibility to tuberculosis. AB - We analysed frequencies of two single-nucleotide polymorphisms (SNP) in the interferon-gamma (IFN-gamma) receptor-1 (IFNGR1) gene promoter (G-611A, T-56C) in tuberculosis patients (n = 244) and compared them with controls (n = 521). These frequencies were not significantly different, whether analysed independently or as haplotypes. Because these SNP affect transcription, the results suggest that the expression of the IFNGR1 gene does not confer susceptibility to disease in patients from Croatia. Further analysis revealed a significant association between the protective (CA)(n) polymorphism (22 repeats, 192 FA(1)), located in the fifth intron of the IFNGR1 gene (+16682), and GT promoter haplotype (-611; 56) that showed the strongest expression capacity. In addition to this cis relationship, the (CA)(22) allele was correlated in trans with an IFN-gamma SNP (IFNG G + 2109A), which might affect the transcription of the IFNG gene. These results suggest that a particular combination of IFNG and IFNGR1 SNP might offer a better protection against tuberculosis in this population. PMID- 16476015 TI - Detection and quantification of herpesviruses in Kostmann syndrome periodontitis using real-time polymerase chain reaction: a case report. AB - BACKGROUND/AIMS: Kostmann syndrome, or severe congenital neutropenia, is an autosomal recessive disease of neutrophil production and is associated with severe periodontal pathology. The aim of this study was to determine whether human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV) contribute to the pathogenesis of Kostmann syndrome periodontitis. METHODS: Supragingival plaque and saliva samples were taken from a 6-year-old boy and his 3-year-old sister suffering from Kostmann syndrome, and from two age- and gender-matched healthy children serving as controls. The samples were taken before and 24 months after periodontal treatment. Real-time polymerase chain reaction (TaqMan Real-Time PCR) assay was used to quantify HCMV and EBV DNA. RESULTS: EBV was detected in baseline samples from the Kostmann syndrome patients but not in samples from the healthy control subjects. HCMV was only detected in the saliva of the boy with Kostman syndrome at baseline. Herpesviruses numbers decreased dramatically in the post-treatment samples. CONCLUSION: EBV and HCMV were detected in the two subjects with Kostmann syndrome periodontitis. The results of the study indicate that nonsurgical treatment of Kostmann syndrome periodontitis can reduce supragingival and salivary herpes viral loads. PMID- 16476016 TI - Use of polymerase chain reaction techniques and sodium dodecyl sulfate polyacrylamide gel electrophoresis for differentiation of oral Lactobacillus species. AB - BACKGROUND/AIMS: The genus Lactobacillus has been associated with dental caries in humans, although it is seldom speciated due to lack of simple and nonlaborious identification methods. A considerable heterogeneity among Lactobacillus species has been demonstrated. The purpose of this study was to develop simple methods combining restriction fragment length polymorphism analysis of polymerase chain reaction (PCR)-amplified 16S rRNA (16S rRNA gene PCR-RFLP) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for the identification of 13 reference strains of Lactobacillus. METHODS: The 16S rRNA gene sequences were amplified by PCR using universal primers and digestion of PCR products with the restriction endonucleases, HpaII and HaeIII. The 16S rRNA gene PCR-RFLP is reproducible and has been proved to be useful for differentiating Lactobacillus strains to species level. Seventy-seven Lactobacillus isolates from a Thai population were used to show the applicability of the identification test. RESULTS: PCR-RFLP alone had limitations, because the RFLP patterns of Lactobacillus casei and Lactobacillus rhamnosus and of Lactobacillus acidophilus and Lactobacillus crispatus showed similar patterns; however, these could be differentiated by SDS-PAGE. Of the 77 isolates, 38 were identified as Lactobacillus fermentum, 25 as L. rhamnosus, 5 as Lactobacillus salivarius, 5 as L. casei, 3 as L. acidophilus and 1 as Lactobacillus plantarum. CONCLUSION: 16S rRNA gene PCR-RFLP, using HpaII and HaeIII, together with SDS-PAGE protein profiles could be an alternative method for the identification of oral Lactobacillus strains to species level, and may be applicable for large-scale studies on the association of Lactobacillus to dental caries. PMID- 16476017 TI - Porphyromonas gingivalis lipids and diseased dental tissues. AB - BACKGROUND/AIM: Porphyromonas gingivalis synthesizes several classes of dihydroceramides and at least one of these lipid classes promotes proinflammatory secretory reactions in gingival fibroblasts as well as alters fibroblast morphology in culture. The purpose of this investigation was to determine whether the dihydroceramide lipids of P. gingivalis are recovered in lipid extracts of subgingival plaque, diseased teeth, and diseased gingival tissue samples. METHODS: Lipids were extracted from P. gingivalis, subgingival plaque, subgingival calculus, teeth laden with gross accumulations of subgingival calculus, and gingival tissue samples obtained from chronic severe periodontitis sites. Lipid samples were analyzed by gas chromatography-mass spectrometry as trimethylsilyl derivatives or by electrospray-mass spectrometry as underivatized products. High-performance liquid chromatography fractions of P. gingivalis lipids and gingival tissue lipids were also analyzed by electrospray-mass spectrometry analysis. RESULTS: P. gingivalis phosphorylated dihydroceramides were recovered in lipid extracts of subgingival plaque, subgingival calculus, calculus contaminated teeth, and diseased gingival tissue samples. However, the distribution of phosphorylated dihydroceramides varied between these samples. CONCLUSION: Subgingival plaque, subgingival calculus, diseased teeth, and gingival tissue are contaminated with phosphorylated dihydroceramides produced by P. gingivalis. The previously reported biological activity of these substances together with the recovery of these lipids at periodontal disease sites argues strongly for their classification as virulence factors in promoting chronic inflammatory periodontal disease. PMID- 16476018 TI - Serum IgG1 and IgG2 antibody responses to Porphyromonas gingivalis in patients with periodontitis. AB - BACKGROUND/AIMS: Protein and carbohydrate antigens of Porphyromonas gingivalis interact with the host to produce antibody of different subclasses. IgG1 and IgG2 antibodies frequently account for approximately 90% of the total serum IgG. This work aimed to investigate serum IgG1 and IgG2 antibody responses of periodontitis patients to protein and carbohydrate-rich antigens of P. gingivalis. METHODS: Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blots of P. gingivalis antigens and proteinase K digested antigens rich in carbohydrates were used to investigate the molecular weight of antigen recognised by serum IgG1 and IgG2. Enzyme-linked immunosorbent assay was used to measure levels of IgG1 and IgG2 antibody to P. gingivalis and radial immunodiffusion was used to estimate the total concentration of IgG1 and IgG2 in serum. RESULTS: Serum IgG antibodies bound to antigens of molecular weights 47, 39 and 32 kDa. Antigen most frequently recognised by both IgG1 and IgG2 antibody had a molecular weight of 47 kDa. Serum IgG2 antibody bound to carbohydrate antigen with a molecular weight of 32 kDa but there was no recognition of carbohydrate antigens by IgG1 antibodies. There was no correlation between the titre of anti-P. gingivalis IgG1 or IgG2 antibody and the total concentration of serum IgG1 or IgG2 antibodies of all specificities. CONCLUSION: Both IgG1 and IgG2 antibodies recognised a dominant antigen of 47 kDa, probably Arg-gingipain. Much of the response to carbohydrate antigen is of the IgG2 subclass. Neither the level of IgG1 nor the IgG2 antibody specific to P. gingivalis was related to the total serum concentration of that antibody. PMID- 16476019 TI - Lethal outcome caused by Porphyromonas gingivalis A7436 in a mouse chamber model is associated with elevated titers of host serum interferon-gamma. AB - BACKGROUND/AIMS: Septic shock caused by gram-negative bacteria has been associated with cytokines produced by hosts. Porphyromonas gingivalis A7436, a disseminating strain, caused septic shock-like symptoms and even animal death in a mouse chamber model. However, P. gingivalis exhibits lower endotoxin activities in its lipopolysaccharide than other typical gram-negative bacteria. In this study, we examined the effects of P. gingivalis lethal infection on host pro inflammatory cytokines production. METHODS: Nude and normal BALB/c mice were infected with a lethal dose of P. gingivalis A7436 using a mouse chamber model. Serum levels of tumor necrosis factor, interleukin (IL)-1beta, IL-12 and interferon-gamma were evaluated. The effects of tumor necrosis factor inhibitor (thalidomide) and anti-interferon-gamma antibody on infection outcomes were examined. RESULTS: All nude mice survived infectious challenge, whereas 100% of normal mice died with abdominal lesions. Bacterial cultures indicated P. gingivalis dissemination to the circulation. Serum levels of tumor necrosis factor, IL-1beta and IL-12 showed no significant differences between nude and normal mice. Thalidomide treatment did not protect normal mice from death but decreased remote lesion occurrence, with concurrent reduced bacterial counts recoverable from blood. There was a 3.5-fold elevation in normal mice serum interferon-gamma titers compared to those of nude mice and anti-interferon-gamma antibody treatment resulted in 100% protection from lethal outcome. CONCLUSION: Lethal outcome following P. gingivalis A7436 infection is T-lymphocyte dependent and involves an increase in systemic interferon-gamma levels. The data further indicate that P. gingivalis transvascular dissemination (bacteremia) alone is not sufficient for lethal outcome. PMID- 16476020 TI - Human cytomegalovirus, Epstein-Barr virus and bone resorption-inducing cytokines in periapical lesions of deciduous teeth. AB - BACKGROUND: A connection of herpesvirus periapical infection with symptomatic and large-size periapical lesions has been recognized in adult patients, but no data exist about a possible involvement of herpesviruses in severe periapical pathosis in children. Herpesviruses have the potential to elicit potent bone resorption inducing cytokines in mammalian cells. AIM: This study aimed to determine the occurrence of human cytomegalovirus and Epstein-Barr virus DNA, and mRNA transcripts of receptor activator of nuclear kappa B ligand (RANKL), osteoprotegerin, core binding factor alpha-1, colony stimulating factor-1, transforming growth factor-beta, and monocyte chemoattractant protein-1 in periapical symptomatic pathosis of deciduous teeth. MATERIAL AND METHODS: Twelve deciduous molar teeth from patients aged 2-8 years were extracted due to severe periapical infection, and granulomatous tissue adherent to the root tip of the extracted teeth was collected using a surgical knife. Non-diseased pulpal tissue, obtained from 12 teeth extracted for orthodontic reasons, served as negative control. Polymerase chain reaction assays were employed to identify herpesvirus DNA and cytokine gene expression, using established polymerase chain reaction primers and procedures. RESULTS: Seven (58%) of the periapical lesions yielded human cytomegalovirus and eight (67%) Epstein-Barr virus. Only one (8%) periapical lesion showed neither human cytomegalovirus nor Epstein-Barr virus. In healthy pulpal tissue, one (8%) specimen demonstrated human cytomegalovirus and another (8%) specimen revealed Epstein-Barr virus. Of the cytokines examined, RANKL expression showed significantly higher occurrence in periapical pathosis than in healthy pulpal tissue (P < 0.040). No relationship was identified between the type of herpesvirus and cytokine expression in the periapical lesions studied. CONCLUSIONS: The present findings provide evidence of a putative role of human cytomegalovirus and Epstein-Barr virus in the pathogenesis of symptomatic periapical pathosis in deciduous teeth. Increased RANKL expression in periapical lesions may be of pathogenetic significance. PMID- 16476021 TI - Molecular analysis of bacteria in asymptomatic and symptomatic endodontic infections. AB - The purpose of the present study was to use terminal restriction fragment length polymorphism analysis and the 16S rRNA gene clone library to investigate the diversity of the microbiota associated with asymptomatic and symptomatic endodontic infections and to compare the bacterial community structure in these two clinical conditions. Samples were taken from asymptomatic endodontic infections associated with chronic periradicular lesions and from symptomatic infections clinically diagnosed as acute abscesses. 16S rRNA genes from DNA isolated from clinical samples were used to construct clone libraries or were subjected to terminal restriction fragment length polymorphism analysis. Sequence analysis of 186 clones revealed 42 taxa; 23 (55%) were uncultivated phylotypes, of which seven were unique to endodontic infections. Clone sequencing and terminal restriction fragment length polymorphism analysis revealed that the most commonly detected taxa were Fusobacterium nucleatum (including terminal restriction fragment types 1 and 2), Peptostreptococcus micros/Peptostreptococcus sp. oral clone AJ062/BS044/FG014, Prevotella species, Dialister species, Mogibacterium species, Lachnospiraceae oral clone 55A-34, Filifactor alocis, Megasphaera sp. oral clone CS025/BS073, and Veillonella sp. oral clone BP1 85/Veillonella dispar/V. parvula. Bacteroides-like sp. oral clone X083/Bacteroidales oral clone MCE7_20 and Dialister sp. oral clone BS016/MCE7_134 were detected only in asymptomatic teeth. On the other hand, F. nucleatum terminal restriction fragment type 2, Prevotella intermedia, Dialister pneumosintes, and some phylotypes were exclusively detected in symptomatic samples. Bacterial profiles of symptomatic endodontic infections generated by terminal restriction fragment length polymorphism analysis were clearly different from those of asymptomatic infections. Overall, the average number of terminal restriction fragments in symptomatic samples was significantly larger than in asymptomatic samples. Molecular analysis of the microbiota associated with symptomatic or asymptomatic endodontic infections indicates that the endodontic bacterial diversity is greater than previously described by culture methods and that the structure of the microbiota differ significantly between asymptomatic and symptomatic infections. PMID- 16476022 TI - Porphyromonas gingivalis gingipains mediate the shedding of syndecan-1 from the surface of gingival epithelial cells. AB - Porphyromonas gingivalis gingipains are thought to be critical virulence factors in periodontitis. Increased serum levels of the soluble ectodomains of surface effectors have been reported to occur during bacterial infections. In the present study, we show that the cell surface proteoglycan syndecan-1 was highly expressed on human gingival epithelial cells. Treatments with P. gingivalis culture supernatants consistently mediated the shedding of syndecan-1 from the surface of epithelial cells. Concomitantly, the amount of soluble syndecan-1 detected in the culture medium increased significantly in a time-dependent manner. However, neither a heat-inactivated supernatant nor a supernatant from a gingipain deficient mutant had a significant effect on syndecan-1 shedding. Such a shedding process may play an important role in the bacterial invasion of periodontal tissue and the modulation of host defences. PMID- 16476023 TI - Colonization of Lactobacillus rhamnosus GG in the oral cavity. AB - BACKGROUND/AIMS: Lactobacillus rhamnosus GG (LGG) is one of the most widely studied probiotic bacterial strain. The benefits of LGG treatment in gastrointestinal disorders are well documented. The aim of the present study was to investigate whether LGG can be detected in the oral cavity after discontinuation of administration of a product prepared with this bacterium. MATERIAL AND METHODS: 56 volunteers consumed Gefilus juice (Valio Ltd, Helsinki, Finland) containing LGG during a 14-day trial period. Saliva samples were collected and cultured onto MRS agar after a clearance period and then daily after a 2-week intervention period for as long as LGG was found. LGG-like colonies were analyzed in saliva samples, identified by characteristic colony morphology, a lactose fermentation test, and PCR with specific primers. RESULTS: LGG was not able to colonize the oral cavity. It could only be temporarily detected. In one female subject, however, whose medical history revealed use of LGG in childhood, the bacterium was detected in all saliva samples taken up to 5 months. (She was excluded from the intervention trial). CONCLUSION: Permanent colonization of LGG in the oral cavity is improbable but seems possible in individual cases. PMID- 16476024 TI - Unmarked gene deletion mutagenesis of gtfB and gtfC in Streptococcus mutans using a targeted hit-and-run strategy with a thermosensitive plasmid. AB - Unmarked gene deletion of the Streptococcus mutans gtfB-gtfC locus was achieved using a thermosensitive plasmid. DNA fragments flanking the locus were amplified by polymerase chain reaction and jointly ligated into pG+host5, which was transformed into S. mutans at 37 degrees C to facilitate integration. A transformant was then grown at 28 degrees C for 60 generations without antibiotics to facilitate excision. Antibiotic sensitive clones appeared at a frequency of about 99% and were analyzed for deletions of gtfB, gtfC and a part of mbrA by the lack of insoluble glucan synthesis, sensitivity to bacitracin, and polymerase chain reaction. Targeted gene deletions occurred at a frequency of 2.5%. PMID- 16476027 TI - Number VII Behcet's disease (Adamantiades syndrome). AB - Behcet's syndrome (BS; Adamantiades syndrome) is the association of the triple symptom complex of recurrent aphthous stomatitis (RAS) with genital ulceration, and eye disease (especially iridocyclitis) though a number of other systemic manifestations may also be seen. BS mainly affects young adult males, and there is an association with HLA-B5 and HLA-B51 (B5101). Features such as arthralgia and leucocytoclastic vasculitis suggest an immune-complex mediated basis, which is supported by finding circulating immune complexes and, although the antigen responsible is unidentified, heat shock proteins have been implicated. An inflammatory disorder, BS is now considered as a systemic vasculitis, characterised by a very wide spectrum of clinical features and by unpredictable exacerbations and remissions. PMID- 16476028 TI - Statins and bone metabolism. AB - 3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) are potent inhibitors of cholesterol biosynthesis. Cholesterol-lowering therapy using statins significantly reduces the risk of coronary heart disease. However, extensive use of statins leads to increases of other undesirable as well as beneficial effects, so-called pleiotropic effects. With respect to these effects, statins augment the expression of bone morphogenetic protein-2, a potent simulator of osteoblast differentiation and its activity, and promote mineralization by cultured osteoblasts, indicating that statins have an anabolic effect on bone. Chronic administration of statins in ovariectomized (OVX) rats modestly increases bone mineral density (BMD) of cancellous bone but not of compact bone. In clinical studies, there are conflicting results regarding the clinical benefits of this therapy for the treatment of osteoporosis. Observational studies suggest an association between statin use and reduction in fracture risk. Clinical trials reported no effect of statin treatment on BMD in hip and spine, and on bone turnover. Statins also may influence oral osseous tissues. Administration of statins in combination with osteoporosis therapy appears to improve alveolar bone architecture in the mandibles of OVX rats with maxillary molar extraction. Statins continue to be considered as potential therapeutic agents for patients with osteoporosis and possibly with periodontal disease. Development of new statins that are more specific and potent for bone metabolism will greatly increase the usefulness of these drugs for the treatment of bone diseases. PMID- 16476029 TI - FGF signalling in craniofacial development and developmental disorders. AB - The Fgf signalling pathway is highly conserved in evolution and plays crucial roles in development. In the craniofacial region, it is involved in almost all structure development from early patterning to growth regulation. In craniofacial skeletogenesis, the Fgf signal pathway plays important roles in suture and synchondrosis regulation. Mutations of FGF receptors relate to syndromatic and non-syndromatic craniosynostosis. The Fgf10/Fgfr2b signal loop is critical for palatogenesis and submandibular gland formation. Perturbation of the Fgf signal is a possible mechanism of palatal cleft. Fgf10 haploinsufficiency has been identified as the cause of autosomal dominant aplasia of lacrimal and salivary glands. The Fgf signal is also a key regulator of tooth formation: in the absence of Fgfr2b tooth development is arrested at the bud stage. Fgfr4 has recently been identified as the key signal mediator in myogenesis. In this review, these aspects are discussed in detail with a focus on the most recent advances. PMID- 16476030 TI - Proinflammatory cytokine levels in saliva before and after treatment of (erosive) oral lichen planus with dexamethasone. AB - OBJECTIVE: To explore the potential of detecting the level of proinflammatory cytokines, tumor necrosis factor-alpha (TNF-alpha), interleukin-1-alpha (IL-1 alpha), IL-6, and IL-8 in whole unstimulated saliva (WUS) in monitoring the therapeutic effects of topical dexamethasone on these salivary cytokines in subjects with erosive oral lichen planus (OLP). STUDY DESIGN: Thirteen definitively diagnosed OLP subjects were enrolled in the study as were 13 age- and sex-matched controls. The OLP subjects were treated with 0.1% dexamethasone oral rinse for 6 weeks. Prior to treatment and at the end of clinical trial, the visual analog scale (VAS) for symptoms was recorded, WUS was collected and these proinflammatory cytokines were analyzed by ELISA. RESULTS: Following the dexamethasone treatment, the levels of TNF-alpha, IL-1-alpha, IL-6, and IL-8 were decreased significantly, and IL-1-alpha and IL-8 were detected at a level without a statistically significant difference from controls. VAS value was decreased significantly and was found to significantly correlate with the decrease in IL-1 alpha and IL-8 levels. CONCLUSIONS: These preliminary results indicate that salivary analysis of NF-kappaB-dependent cytokines may be applied to monitoring the therapeutic response of OLP. PMID- 16476032 TI - Manifestations of the tongue in Neurofibromatosis type 1. AB - OBJECTIVE: The aim of this study is to analyse alterations of the tongue and the correlation between these lesions and different types of tumor. SUBJECTS AND METHODS: A total of 258 cases (131 females, 127 males) of neurofibromatosis type 1 were screened between 1994 and 2004 in our Dermatology Department. All patients included in this study have NF1, as defined by the NIH Consensus Conference. Three cases of neurofibromas of the tongue in patients with neurofibromatosis type were reported. RESULTS: Our patients showed nodular lesions on the tongue, related to neurofibromas in two patients and plexiform neurofibroma in one patient, respectively. Clinical and hystopatological findings were useful in distinguishing between neurofibromas and other soft tissue tumors. An increased prevalence of malignancy has been documented in patients affected by neurofibromatosis type 1. Changes in the size of a pre-existing mass, compression, or infiltration of the adjacent structures indicate malignant degeneration. Histological and clinical evaluation should be performed in order to choose the most appropriate treatment strategy for these patients. CONCLUSION: The oral manifestations of NF are well-documented but may not be at the forefront of the clinician's mind in the differential diagnosis of intra-oral swellings. PMID- 16476031 TI - Prevalence of erosive tooth wear and associated risk factors in 2-7-year-old German kindergarten children. AB - OBJECTIVES: The aims of this study were to (1) investigate prevalence and severity of erosive tooth wear among kindergarten children and (2) determine the relationship between dental erosion and dietary intake, oral hygiene behaviour, systemic diseases and salivary concentration of calcium and phosphate. MATERIALS AND METHODS: A sample of 463 children (2-7 years old) from 21 kindergartens were examined under standardized conditions by a calibrated examiner. Dental erosion of primary and permanent teeth was recorded using a scoring system based on O'Sullivan Index [Eur J Paediatr Dent 2 (2000) 69]. Data on the rate and frequency of dietary intake, systemic diseases and oral hygiene behaviour were obtained from a questionnaire completed by the parents. Unstimulated saliva samples of 355 children were analysed for calcium and phosphate concentration by colorimetric assessment. Descriptive statistics and multiple regression analysis were applied to the data. RESULTS: Prevalence of erosion amounted to 32% and increased with increasing age of the children. Dentine erosion affecting at least one tooth could be observed in 13.2% of the children. The most affected teeth were the primary maxillary first and second incisors (15.5-25%) followed by the canines (10.5-12%) and molars (1-5%). Erosions on primary mandibular teeth were as follows: incisors: 1.5-3%, canines: 5.5-6% and molars: 3.5-5%. Erosions of the primary first and second molars were mostly seen on the occlusal surfaces (75.9%) involving enamel or enamel-dentine but not the pulp. In primary first and second incisors and canines, erosive lesions were often located incisally (51.2%) or affected multiple surfaces (28.9%). None of the permanent incisors (n = 93) or first molars (n=139) showed signs of erosion. Dietary factors, oral hygiene behaviour, systemic diseases and salivary calcium and phosphate concentration were not associated with the presence of erosion. CONCLUSIONS: Erosive tooth wear of primary teeth was frequently seen in primary dentition. As several children showed progressive erosion into dentine or exhibited severe erosion affecting many teeth, preventive and therapeutic measures are recommended. PMID- 16476033 TI - H3 and H3.3 histone mRNA amounts and ratio in oral squamous cell carcinoma and leukoplakia. AB - Histone variants (e.g. H3) play an important role in chromatin structure and gene expression regulation of normal cells. Aims of this study were to: (1) estimate H3 and H3.3 histone mRNA expressions and their ratio in oral squamous cell carcinoma (OSCC) and oral leukoplakia (OL); (2) investigate whether H3 and H3.3 variants could play a role in the pathogenesis of OSCC and OL, also conditionally to HPV infection, age, gender, and main habits (tobacco smoking and alcohol drinking) in human beings studied. Twenty-three cases of OSCC and 20 cases of OL were examined in lesion site (LS) and juxtaposed clinically undamaged site (JUS) by RT-PCR for H3 and H3.3 histone mRNA; 13 healthy oral mucosa samples (HS) were investigated in a single site as controls. HPV DNA presence was investigated in the respective exfoliated oral mucosa cells by nested PCR (nPCR: MY09-MY11/GP5 GP6). The data showed that both H3 and H3.3 histone mRNA crude concentrations are higher in OSCC (LS = 2901 +/- 459 ng of H3; JUS = 2699 +/- 658 ng of H3; LS = 3190 +/- 411 ng of H3.3; JUS = 2596 +/- 755 ng of H3.3) than those in OL (LS = 2095 +/- 349 ng of H3; JUS = 2192 +/- 897 ng of H3; LS = 2076 +/- 911 ng of H3.3; JUS = 1880 +/- 654 ng of H3.3) and in HS (2579 +/- 959 ng of H3; 2300 +/- 758 ng of H3.3), although not reaching any statistical significance. Interestingly, ratio of H3/H3.3 mRNA amounts decrease both in OSCC (0.99) and OL (1.009) vs HS (1.121). No association was found for H3 and H3.3 histone mRNA expressions in OSCC and OL with respect to HPV infection and the social-demographical variables considered (P > 0.2). The overall higher expression of H3.3 in damaged tissues up to the ratio inversion in OSCC especially in HPV+ alcohol drinkers (60.0%) represents the most interesting finding, in consideration of the proven ability of alcohol to act as permeability enhancer of human oral mucosa, to alter the mucosal structure and by this dynamics could favour the penetration through the epithelial layers of HPV. PMID- 16476034 TI - Toxicity and biodistribution of a first-generation recombinant adenoviral vector, in the presence of hydroxychloroquine, following retroductal delivery to a single rat submandibular gland. AB - OBJECTIVE: We examined the toxicity and biodistribution associated with a single administration of a first-generation, serotype 5, adenoviral vector encoding human growth hormone (hGH; AdCMVhGH) to a single rat submandibular gland in the presence of hydroxychloroquine (HCQ). Previously, we showed that hGH is primarily secreted into saliva (approximately ninefold serum level) when expressed as a transgene in salivary glands (e.g. Baum et al, 1999), but administration of HCQ substantially increases the hGH levels secreted into the bloodstream (Hoque et al, 2001). A potential application of this observation is for patients with adult hGH deficiency. METHODS: Six groups of male and female adult rats (n = 12 each) were studied, with zero to 1.5 x 10(11) particles of AdCMVhGH, +/-HCQ, administered retroductally. Multiple clinical and pathological parameters, as well as vector tissue distribution, were assessed. RESULTS: All animals survived until the scheduled day of sacrifice, and essentially no untoward events were observed clinically or at gross necropsy. We observed no vector-related effects on clinical hematology evaluations and a single, transient significant change on clinical chemistry evaluations (increased serum globulin levels). Three days after AdCMVhGH administration, the vector distributed to all tissues analyzed with the exception of gonads and heart. By day 29, most organs, other than the targeted and contralateral submandibular glands, were negative for the presence of vector. On day 3, none of the animals tested positive for the presence of replication competent adenovirus in either their blood or saliva. CONCLUSION: Salivary gland delivery of AdCMVhGH +/-HCQ appears associated with limited toxicity in rats. PMID- 16476035 TI - Oral health related quality of life is affected by disease activity in Behcet's disease. AB - OBJECTIVES: This study aimed to investigate oral and general health related quality of life (QoL) in patients with Behcet's Disease (BD) and to assess the performance of Turkish versions of oral health related quality questionnaires. SUBJECTS AND METHODS: Ninety-four BD patients, 24 patients with recurrent aphthous stomatitis (RAS), 113 healthy controls (HC) and 44 dental patients were investigated. QoL was assessed by oral health impact profile-14 (OHIP-14), oral health related quality of life (OHQoL) and short form-36 (SF-36) questionnaires. RESULTS: OHQoL, OHIP-14 and SF-36 subscale scores were significantly worse in patients with BD compared with those in HC (P < 0.05). Both OHIP-14 and OHQoL scores were significantly worse in active patients compared with inactives in BD and RAS (P < 0.05). Scores of SF-36 Role physical, Role emotional and Vitality were also lower in active patients than in inactives in BD (P < 0.05). Scores of OHIP-14 and OHQoL were significantly worse in patients treated with colchicine compared with those treated with immunosuppressives (P < 0.05). CONCLUSIONS: Both oral and general QoL was impaired in BD and associated with disease activity and treatment modalities. Translated Turkish versions of OHIP-14 and OHQoL were also observed to be valid and reliable questionnaires for further studies. PMID- 16476036 TI - Burning mouth syndrome: a retrospective study investigating spontaneous remission and response to treatments. AB - OBJECTIVE: The aim of this investigation was to evaluate the spontaneous remission rate of burning mouth syndrome (BMS) in a group of subjects suffering from this syndrome. SUBJECTS AND METHODS: The medical records of BMS patients attending the Unit of Oral Medicine (1995-2002) were reviewed. The patients with a follow-up period of at least 18 months were then contacted over phone and interviewed using a structured ad hoc questionnaire to record their current symptoms and data about their treatment responses to the therapies. RESULTS: Forty-eight women and five men with a mean age of 67.7 years (range 33-82 years) were included in the study (mean duration of BMS 5.5 years, s.d. +/-1.9 years, mean follow-up period of 56 months). As a consequence of different treatments, 26 patients (49.0%) reported no change in oral symptoms, 15 (28.3%) moderate improvement and 10 (18.9%) a worsening of oral complaints. Only two patients (3.7%) reported a complete spontaneous remission of oral symptoms without any treatment. CONCLUSIONS: In this study, a complete spontaneous remission was observed in 3% of the patients within 5 years after the onset of BMS. A moderate improvement was obtained in <30% of the subjects. PMID- 16476037 TI - Effect of phenytoin on collagen accumulation by human gingival fibroblasts exposed to TNF-alpha in vitro. AB - OBJECTIVE: Tumor necrosis factor (TNF)-alpha is associated with chronic gingival inflammation and reported to induce gingival overgrowth (GO), while phenytoin (PHT) is also known to be a causative agent of GO. We examined the synergistic effect of PHT and TNF-alpha on collagen metabolism in human gingival fibroblasts (HGFs). MATERIALS AND METHODS: HGFs were cultured with TNF-alpha and PHT. Quantitative real-time RT-PCR was employed to determine the mRNA levels for collagen, matrix metalloproteinases (MMPs), tissue inhibitors of metalloproteinases (TIMPs) and integrin subunits. Cellular collagen endocytosis was determined using a flow-cytometry. RESULTS: The proliferation of HGFs was not affected by TNF-alpha or PHT individually, whereas both synergistically increased collagen accumulation in HGFs. Further, collagen mRNA expression was not increased by TNF-alpha or PHT, although together they markedly prevented cellular collagen endocytosis, associated with the suppression of alpha2beta1-integrin mRNA expression. The mRNA expression of MMP-1 and-2 was suppressed by PHT, while TIMP-1 mRNA expression was enhanced by both TNF-alpha and PHT. CONCLUSION: Our results suggest that TNF-alpha and PHT together cause impaired collagen metabolism by suppression of enzymatic degradation with MMPs/TIMP-1 and integrin mediated endocytosis. These synergistic effects may also be involved in TNF-alpha and PHT-induced collagen accumulation, leading to GO. PMID- 16476038 TI - Expression of bone morphogenetic proteins and their associated molecules in ameloblastomas and adenomatoid odontogenic tumors. AB - OBJECTIVE: To further clarify the roles of regulators of embryonic development, bone morphogenetic protein (BMPs) and their associated molecules, in oncogenesis and cytodifferentiation of odontogenic tumors, the expression of these regulator molecules were analyzed in epithelial odontogenic tumors as well as in tooth germs. MATERIALS AND METHODS: Tooth germs, ameloblastomas, adenomatoid odontogenic tumors, and malignant ameloblastomas were examined by RT-PCR and immunohistochemistry for detection of BMP-2, -4, -7, BMP receptors I and II (BMPR I, BMPR-II), core-binding factor alpha1 (CBFA1), and osterix. RESULTS: mRNA expression of BMPs, BMPRs, CBFA1, and osterix was detected in all odontogenic tissues. Immunohistochemical reactivity for BMPs, BMPRs, and CBFA1 was detected in both epithelial and mesenchymal cells of tooth germs and epithelial odontogenic tumors. BMPs and BMPRs were evidently expressed in odontogenic epithelial cells in tooth germs and epithelial odontogenic tumors. Acanthomatous ameloblastomas showed increased BMP-7 reactivity in keratinizing cells. Nuclear CBFA1 expression was detected scatteredly in odontogenic epithelial cells in normal and neoplastic odontogenic tissues, as well as in some mesenchymal cells in tooth germs and in some stromal cells in epithelial odontogenic tumors. Ameloblastic carcinomas showed low reactivity for BMPs, BMPRs, and CBFA1. CONCLUSION: BMPs and their associated molecules might play a role in cytodifferentiation of normal and neoplastic odontogenic epithelium via epithelial-mesenchymal interactions. PMID- 16476039 TI - Mechanical stress induces expression of cytokines in human periodontal ligament cells. AB - OBJECTIVE: Periodontal tissue has a unique structure in that the human periodontal ligament (hPDL) lies between the hard tissues of cementum and alveolar bone. Although the role of cytokines in hPDL function is not clearly understood, we investigated the effect of mechanical stress as hydrostatic pressure (HP) on cytokine expression in hPDL cells. MATERIALS AND METHODS: The hPDL cells were obtained from a healthy maxillary third molar. After the third to fourth passage, the cells were exposed to HP ranging from 1 to 6 MPa as previously described. Total RNA was extracted and the expression of cytokine mRNA was determined by RT-PCR. RESULTS: The exposure to 6 MPa of HP caused no morphological changes of hPDL cells, and did not affect the cellular viability. No expression of IL-1beta, IL-6, IL-8, TNF-alpha, receptor activator of NF lambdaB (RANK), receptor activator of NF-lambdaB ligand (RANKL), or osteoprotegerin mRNA was observed in the control cells under atmospheric pressure, whereas, in hPDL cells treated with HP, a pressure-dependent enhancement of IL-6, IL-8, RANKL, and OPG mRNA expression was observed between 10 and 60 min after the exposure to HP. CONCLUSION: These results suggest that hPDL cells may play a role in the production of cytokines in response to mechanical stress in vivo. PMID- 16476040 TI - The oral health status of dentate patients with chronic renal failure undergoing dialysis therapy. AB - OBJECTIVE: The aim of this study was to compare the oral health status of chronic renal failure (CRF) patients on renal replacement therapy with a matched reference population. DESIGN: Cross-sectional study. SUBJECTS: Forty-two dentate CRF patients--aged 25-52 years old--were matched with a reference group of 808 dentate subjects. METHODS: The oral health was assessed using decayed missing filled (DMF) indices, simplified oral hygiene index and periodontal status. An oral health questionnaire was used to assess self-reported dental problems. Student t-tests and chi-square tests were performed to compare the CRF patients with the controls. RESULTS: All index-scores in the CRF patients were comparable with the controls except for number of teeth covered with calculus that was significantly higher (P < 0.05) in CRF patients (4.1 +/- 2.6) than in controls (3.0 +/- 2.9). The self-reported oral health questionnaire revealed a trend for increased temporomandibular complaints in CRF patients (16.7%vs 5.7% in controls; P = 0.06) as well as bad taste (31.0%vs 6.8% in controls, P = 0.08). CONCLUSIONS: For most dental aspects, the oral health of CRF patients is comparable with controls. PMID- 16476041 TI - Mandible analysis in sex steroid-deficient rats. AB - The aim of this study was to evaluate, by morphologic techniques, the effects of sex steroid deficiency on mandible bone remodeling of female rats, in groups of different experimental periods and to compare the results with 90-day orquiectomized males. Female and male Wistar rats, 3 months old, were divided into experimental groups and at the end of each experimental period were killed, and mandibles were extracted. The left mandibles were prepared with rote technique bone and examined by a light microscope. Morphological analyses of the mandibles demonstrated resorption signals in the alveolar bone, after 30 days in ovariectomized females, but it was more intense 90 days after castration. The orquiectomized group exhibited some signals of resorption similar to the ovariectomized group of 60 days. Morphometric analysis of alveolar bone thickness in females after 60 days was in agreement with morphological results. However, the analysis of periodontal ligament thickness did not show any significant difference. There were variations in sexual hormone deficiency in the mandibles of males and females and they seemed to be more precocious in ovariectomized than in orquiectomized rats. It is important for a health professional to have knowledge about bone metabolism to improve the quality of life of postmenopaused and old people. PMID- 16476042 TI - Effects of alendronate and hormone replacement therapy, alone and in combination, on saliva, periodontal conditions and gingival crevicular fluid matrix metalloproteinase-8 levels in women with osteoporosis. AB - OBJECTIVE: To compare the effects of hormone replacement therapy (HRT), alendronate and their combination on oral health of elderly postmenopausal women with osteoporosis. MATERIALS AND METHODS: Sixty patients, aged 65-80 years (mean 71 years), with a T-score of bone mineral density of -2.5 s.d. or less at either the lumbar spine or the femoral neck, were randomized to receive 2 mg of estradiol plus 1 mg norethisterone acetate (HRT) (n = 20), 10 mg of alendronate (n = 18), or their combination (n = 22) for 2 years. Periodontal and oral status and mouth symptoms were recorded, and salivary analyses made at the beginning and at the end of the study. Gingival crevicular fluid (GCF) matrix metalloproteinase (MMP-8) levels were determined to address destructive events in periodontal tissue. RESULTS: Resting salivary flow rate decreased by 19% (P < 0.05), and GCF MMP-8 tended to increase in the alendronate group. None of the regimens affected subjective feelings of dry or burning mouth. There were no significant changes in dental or periodontal status, stimulated flow rate or composition of saliva during the study. CONCLUSIONS: Alendronate decreased resting salivary flow rate but otherwise HRT or alendronate separately or in combination had no effect on oral health in elderly women with osteoporosis. PMID- 16476043 TI - Opinions and attitudes of the UK's GDPs and specialists in oral surgery, oral medicine and surgical dentistry on oral cancer screening. AB - OBJECTIVE: To survey two broad areas of oral cancer awareness and management of patients at risk of oral cancer by specialists in oral surgery, oral medicine, surgical dentistry and general dental practitioners (GDPs) in the UK. The first of these included knowledge and awareness of aetiological factors, changing patterns of disease, and screening/detection programmes including their effectiveness. The second included oral cancer detection methods, advice on avoidance of high-risk activity and self-examination, and referral pattern of GDPs. DESIGN AND METHOD: A pretested, 44-item questionnaire, a covering letter, a brief outline of the research protocol and return, stamped envelope were mailed in March 2003. A sample of 200 GDPs whose names were obtained from the General Dental Council's main list and 305 dental specialist names obtained from specialist's list in surgical dentistry, oral medicine and oral surgery were selected randomly. Information on oral cancer awareness and practice, screening practice and education was obtained. RESULTS: The response rate was 66.9%. The knowledge of the dental specialists was consistent with that in reports of current aetiological studies on oral cancer. However there were gaps in the GDP's knowledge and ascertainment of oral cancer risk factors. Over 70% of the dental specialists provided counselling advice on the risks of tobacco and alcohol habits compared with 41.2% of GDPs. More GDPs (52.4%) than specialists (35.4%) believed that oral cancer screening on a national basis would be effective in decreasing the mortality of oral cancer. Over 95% of all respondents used a visual examination for oral cancer screening and 89.9% of all respondents strongly believed that visual screening is effective in the early detection of oral cancer. CONCLUSION: The results showed that GDPs had knowledge gaps in their awareness of oral cancer risk factors and the application of preventive measures. Most dental health providers in the UK perform visual screening of the oral mucosa for their patients. Opinion was equivocal as to whether a nationally based screening programme similar to cervical cancer would be effective in improving the mortality and morbidity of oral cancer. PMID- 16476044 TI - Adenomatoid dentinoma or adenomatoid odontogenic hamartoma: what is the better term to denominate this uncommon odontogenic lesion? AB - We report two cases of an uncommon odontogenic lesion, previously described as adenomatoid dentinoma. They were well-circumscribed unilocular radiolucent lesions exhibiting discrete radiopacities, located in the left mandibular third molar region. Microscopically they were composed of odontogenic hard and soft tissues, similar to a dental germ. Dental papilla and dentin were easily identified. Odontogenic epithelium formed adenomatoid-like structures, and by scanning electron microscopy a layer of enamel was seen in contact with the dentin. Based on these clinical, radiographic, histological and electron microscopical features we proposed the diagnosis of adenomatoid odontogenic hamartoma. Treatment consisted of surgical removal, and no recurrence was observed. In our opinion all similar cases previously reported pertain to the same spectrum of this lesion and thus should be named as suggested above. Moreover, ultrastructural observations using 5 microm sections can be useful to better characterize the presence of hard tissues. PMID- 16476045 TI - Fibrodysplasia ossificans progressiva. An unusual cause of restricted mandibular movement. AB - A 9-year-old girl is presented who was initially misdiagnosed and finally diagnosed with fibrodysplasia ossificans progressiva only after presentation with progressive limitation of her mouth opening. The clinical, histopathological, and molecular biological aspects of this uncommon disorder will be discussed. Furthermore, dental and surgical guidelines will be described. PMID- 16476046 TI - Dental findings and rehabilitation in familial osteodysplasia (Anderson type): a case report. AB - Familial osteodysplasia is a disorder of osteogenesis with an autosomal recessive pattern of inheritance which predominantly affects facial bones. No recent case had been reported, particularly from a dental point of view since the syndrome was first described by Anderson et al (JAMA 1972;220:1687-93). A 23-year-old male with familial osteodysplasia was presented in maxillofacial and dental aspects with clinical and radiological manifestations including malocclusion, abnormal teeth alignment, impacted teeth, shape disturbances including uncompleted coronal formation, root shortening with bulbous form, high angled mandible and elongation of the corpus of mandible. Recognition of the syndromal features prior to any dental intervention is of paramount importance because of increased inclination to spontaneous mandibular fractures. Hence, no surgical intervention was performed for impacted teeth. Following the extractions of severely mobile teeth, a definitive restoration was fabricated as distal-extension removable partial dentures with conus crown telescopic system. The aesthetic and functional outcome was satisfactory for the patient. In conclusion, dentists appear to play an important role in the recognition of familial osteodysplasia, based on maxillofacial and dentoalveolar findings. Awareness of the syndromal features, especially of spontaneous fractures, would detect the limitations for dental interventions and treatment planning. PMID- 16476047 TI - Primary parotid tuberculosis: varied clinical presentations. AB - Parotid gland tuberculosis is rare and may present in different clinical forms. We present three cases of tuberculosis of the parotid gland that presented to us with different clinico-pathological appearances. The cases were diagnosed on the basis of histopathological evaluation and fine needle aspiration cytology. All the patients responded to four-drug antitubercular chemotherapy. PMID- 16476048 TI - Imbalance of regulatory T cells in human autoimmune diseases. AB - The breakdown of mechanisms assuring the recognition of self and non-self is a hallmark feature of autoimmune diseases. In the past 10 years, there has been a steadily increasing interest in a subpopulation of regulatory T cells, which exert their suppressive function in vitro in a contact-dependent manner and preferentially express high levels of CD25 and forkhead and winged-helix family transcription factor forkhead box P3 (FOXP3) (TREGs). Recent findings of changed prevalences and functional efficiencies indicate that these TREGs play a unique role in autoimmune diseases. Clinical findings in patients with mutated FOXP3 genes and a specific polymorphism in the promotor region of FOXP3 also support the role of FOXP3 as a 'master control gene' in the development and functioning of TREGs. Both altered generation of TREGs and insufficient suppression of inflammation in autoimmune diseases are considered to be crucial for the initiation and perpetuation of disease. TREG-related somatic cell therapy is considered as an intriguing new intervention to approach autoimmune diseases. PMID- 16476050 TI - Vitamin D receptor is required to control gastrointestinal immunity in IL-10 knockout mice. AB - The vitamin D receptor (VDR) is a nuclear receptor expressed in a number of different cells of the immune system. This study was performed to determine the effect of VDR deficiency on immune function and inflammation of the gastrointestinal tract in a model of inflammatory bowel disease, namely interleukin-10 (IL-10) knockout mice. IL-10 knockout mice were generated which either could or could not respond to vitamin D (double IL-10/VDR knockout; DKO). The distribution and function of lymphocytes in both the primary and secondary lymphoid organs were compared and determined as a function of the severity of intestinal inflammation. DKO mice had normal thymic development and peripheral T cell numbers at 3 weeks of age, but a week after intestinal disease was detected the thymus was dysplastic with a reduction in cellularity. The atrophy was coupled with increased apoptosis. The spleen weight of DKO mice increased as a result of the accumulation of red blood cells; however, there was a 50% reduction in the numbers of T and B cells. Conversely, the mesenteric lymph nodes were enlarged and contained increased numbers of lymphocytes. The T cells from DKO mice were of a memory phenotype and were hyporesponsive to T-cell receptor stimulation. Colitis in the DKO mice was associated with local and high expression of IL-2, interferon-gamma, IL-1beta, tumour necrosis factor-alpha and IL-12. The primary and secondary lymphoid organs in DKO mice are profoundly altered as a consequence of the fulminating inflammation in the gastrointestinal tract. VDR expression is required for the T cells and other immune cells to control inflammation in the IL-10 KO mice. PMID- 16476049 TI - Dealing with the family: CD147 interactions with cyclophilins. AB - CD147 is a widely expressed plasma membrane protein that has been implicated in a variety of physiological and pathological activities. It is best known for its ability to function as extracellular matrix metalloproteinase inducer (hence the other name for this protein, EMMPRIN), but has also been shown to regulate lymphocyte responsiveness, monocarboxylate transporter expression and spermatogenesis. These functions reflect multiple interacting partners of CD147. Recently, interaction of CD147 with proteins of the cyclophilin family has been demonstrated and activity of CD147 as a signalling receptor to extracellular cyclophilins A and B has been shown. Given that extracellular cyclophilins are potent chemotactic agents for various immune cells, further studies of the role of cyclophilin-CD147 interaction in inflammation followed. They demonstrated that agents targeting CD147 or cyclophilin had a significant anti-inflammatory effect in animal models of acute or chronic lung diseases and rheumatoid arthritis. Here, we review the current knowledge about interactions between CD147 and cyclophilins. PMID- 16476051 TI - Luminal bacterial antigen-specific CD4+ T-cell responses in HLA-B27 transgenic rats with chronic colitis are mediated by both major histocompatibility class II and HLA-B27 molecules. AB - Rats transgenic (TG) for the human major histocompatibility complex (MHC) class I HLA-B27 and beta2-microglobulin genes develop chronic colitis under specific pathogen-free (SPF) but not sterile (germ-free, GF) conditions. We investigated the role of antigen-presenting molecules involved in generating immune responses by CD4+ mesenteric lymph node (MLN) cells from colitic HLA-B27 TG rats to commensal enteric micro-organisms. All TG MLN cells expressed HLA-B27. A higher level of MHC class II was expressed on cells from TG rats, both SPF and GF, compared to non-TG littermates. In contrast, rat MHC class I expression was lower on TG than non-TG cells. Both TG and non-TG antigen presenting cells (APC) pulsed with caecal bacterial antigens induced a marked interferon-gamma (IFN-gamma) response in TG CD4+ T lymphocytes but failed to stimulate non-TG cells. Blocking MHC class II on both TG and non-TG APC dramatically inhibited their ability to induce TG CD4+ T cells to produce IFN-gamma. Blocking HLA-B27 on TG APC similarly inhibited IFN-gamma responses. When the antibodies against MHC class II and HLA B27 were combined, no APC-dependent IFN-gamma response was detected. These data implicate both native rat MHC class II and TG HLA-B27 in CD4+ MLN T-cell IFN gamma responses to commensal enteric microflora in this colitis model. PMID- 16476052 TI - Penetratin tandemly linked to a CTL peptide induces anti-tumour T-cell responses via a cross-presentation pathway. AB - Recently there has been increasing evidence to suggest that membrane translocating peptides enter cells by a receptor-dependent pathway. There have been some studies on the mechanism of major histocompatibility complex (MHC) class I presentation of membrane translocating peptides incorporating cytotoxic T lymphocyte epitopes. However, these have been on different cell lines and only a limited number of inhibitors of the antigen presentation pathway were used. Herein, we demonstrate a comprehensive study utilizing a full spectrum of inhibitors to various pathways of MHC class I to elucidate the mechanism of the membrane translocating peptide, penetratin from Antennapedia (Int). It is clear that Int, RQIKIWFQNRRMKWKK when tandemly linked to a cytotoxic T lymphocyte peptide of ovalbumin, SIINFEKL (IntSIIN) is endocytosed via phagocytosis or macropinocytosis by dendritic cells in an ATP-dependent manner and is processed by a proteasome- and tapasin-independent pathway for presentation and loading to MHC class I molecules. In addition, the majority of antigen is taken up by negatively charged receptors. IntSIIN activates T cells in vitro and in vivo and protects mice against challenge with an ovalbumin-expressing tumour. PMID- 16476054 TI - Cytotoxic herpes simplex type 2-specific, DQ0602-restricted CD4 T+-cell clones show alloreactivity to DQ0601. AB - Alloreactivity is one of the most serious problems in organ transplantation. It has been hypothesized that pre-existing alloreactive T cells are actually cross reacting cells that have been primed by the autologous major histocompatibility complex (MHC) and a specific peptide. CD8+ cytotoxic T lymphocytes that are alloreactive and recognize a virus-peptide that is presented by the autologous MHC have been reported. Here we demonstrate a cross-reactivity that exists between DQ0602 restricted, herpes simplex type 2 VP16 40-50 specific CD4+ T-cell clones, which can be alloreactive to DQ0601. Though most of the DQ0602 restricted T-cell clones we isolated from two different donors were not alloreactive, weakly cross-reacting T-cell clones could be isolated from both donors. Two strongly cross-reacting T-cell clones with high affinity interaction of their T-cell receptor (TCR) with both DQ0602/VP16 40-50 and DQ0601 could be isolated from one donor. DNA sequencing of the a fragment of the Vbeta gene used in their TCR confirmed that these two T cells indeed are two independent clones. These clones are cytotoxic and produce cytokines of a T helper 2-like pattern. Possible implications in a DR-matched transplantation setting are discussed. PMID- 16476053 TI - Differential modulatory effects of annexin 1 on nitric oxide synthase induction by lipopolysaccharide in macrophages. AB - Annexin-1 (ANXA1) is a glucocorticoid-regulated protein that modulates the effects of bacterial lipopolysaccharide (LPS) on macrophages. Exogenous administration of peptides derived from the N-terminus of ANXA1 reduces LPS stimulated inducible nitric oxide synthase (iNOS) expression, but the effects of altering the endogenous expression of this protein are unclear. We transfected RAW264.7 murine macrophage-like cell lines to over-express constitutively ANXA1 and investigated whether this protein modulates the induction of iNOS, cyclooxygenase-2 (COX-2) and tumour necrosis factor-alpha (TNF-alpha) in response to LPS. In contrast to exogenous administration of N-terminal peptides, endogenous over-expression of ANXA1 results in up-regulation of LPS-induced iNOS protein expression and activity. However, levels of iNOS mRNA are unchanged. ANXA1 has no effect on COX-2 or TNF-alpha production in response to LPS. In experiments to investigate the mechanisms underlying these phenomena we observed that activation of signalling proteins classically associated with iNOS transcription was unaffected. Over-expression of ANXA1 constitutively activates extracellular signal regulated kinase (ERK)-1 and ERK-2, components of a signalling pathway not previously recognized as regulating LPS-induced iNOS expression. Inhibition of ERK activity, by the inhibitor U0126, reduced LPS induced iNOS expression in our cell lines. Over-expression of ANXA1 also modified LPS-induced phosphorylation of the ERK-regulated translational regulation factor eukaryotic initiation factor 4E. Our data suggest that ANXA1 may modify iNOS levels by post-transcriptional mechanisms. Thus differential effects on iNOS expression in macrophages are seen when comparing acute administration of ANXA1 peptides versus the chronic endogenous over-expression of ANXA1. PMID- 16476055 TI - Cytotoxic T-lymphocyte antigen-4 inhibits GATA-3 but not T-bet mRNA expression during T helper cell differentiation. AB - Naive CD4+ T-cell differentiation to T helper 1 (Th1) and Th2 cells is dependent on T-bet and GATA-3 factors, respectively. T-bet and GATA-3, indeed, through chromatin remodelling allow transcriptional activation of Ifngamma and Th2 cytokine (Il4, Il5, Il13) genes, respectively. We investigated the effects of the negative costimulatory receptor cytotoxic T-lymphocyte antigen-4 (CTLA-4) on GATA 3 and T-bet mRNA expression and Th cell differentiation in mouse naive CD4+ T cells. Our results show that CTLA-4 inhibits GATA-3 mRNA expression and Th2 cell differentiation. At variance, CTLA-4 does not affect T-bet mRNA expression and Th1 cell differentiation. GATA-3 mRNA expression is inhibited when CD4+ cells are stimulated under both neutral (i.e. absence of cytokines) and Th2-polarizing (i.e. presence of interleukin (IL)-4) conditions, the effect being larger under the latter condition. Hence CTLA-4 might affect the IL-4/signal transducer and activator of transcription-6 (STAT6) pathway leading to GATA-3 mRNA up regulation. We found, indeed, that CTLA-4 engagement inhibits STAT6 activation leaving unaffected the STAT6 protein level. Moreover, CTLA-4 engagement drastically inhibits IL-4Ralpha mRNA and protein up-regulation under Th2 polarizing conditions. Thus, CTLA-4 exerts a tight control on Th2 cell differentiation by negatively regulating both the CD3/CD28 and the IL-4/STAT6 pathways. PMID- 16476056 TI - Induction of autoimmunity by immunization with hapten-modified hen egg lysozyme in hen egg lysozyme-transgenic mice. AB - To understand the mechanism of autoimmunity induction, hen egg lysozyme (HEL) transgenic (Tg) C57BL/6 (B6) mice were immunized with HEL or phosphorylcholine conjugated HEL (PC-HEL). Repeated immunization of HEL-Tg mice with native HEL failed to induce the antibody response against HEL. However, immunization with PC HEL generated a significant anti-HEL antibody response. Immunization of the Tg mice with dominant (HEL(74-88)) or cryptic (HEL(47-61)) T-cell epitope peptide stimulated the corresponding T-cell response and similarly yielded the anti-HEL antibody response. Predominance of immunoglobulin G1 (IgG1) anti-HEL antibody response in the HEL-Tg mice and preferential IL-4 production by HEL-specific T cells suggested the dependency of the antibody response to the presence of T helper 2. HEL-Tg mice received HEL-primed B6 T cells, but not HEL-primed Tg T cells, were able to generate anti-HEL antibody response following PC-HEL immunization. The pattern and the level of epitope peptides generated by splenic antigen-presenting cells indicated that PC-HEL results in much more efficient processing as compared to HEL. These results strongly suggest that the enhancement of antigen processing by hapten (PC) conjugation to the antigen facilitates more efficient stimulation of T cells reactive to self antigen, HEL in HEL-Tg mice resulting in the production of anti-self HEL antibody. PMID- 16476057 TI - Epstein-Barr virus infection negatively impacts the CXCR4-dependent migration of tonsillar B cells. AB - The primary Epstein-Barr virus (EBV) infection occurs in the oropharynx, where the virus infects B cells and subsequently establishes latency in the memory B cell compartment. EBV has previously been shown to induce changes in the cell surface expression of several chemokine receptors in cell lines and the transfection of EBNA2 or LMP1 into a B-cell-lymphoma-derived cell line decreased the expression of CXCR4. We show that in vitro EBV infection reduces the expression of CXCR4 on primary tonsil B cells already 43 hr after infection. Furthermore, EBV infection affects the chemotactic response to stromal cell derived factor (SDF-1)alpha/CXCL12, the ligand for CXCR4, with a reduction of SDF 1alpha-induced migration. To clarify whether this reduced migration is EBV specific or a consequence of cell activation, tonsillar B cells were either infected with EBV, activated with anti-CD40 and interleukin-4 (IL-4) or kept in medium. Activation by anti-CD40 and IL-4 decreased the CXCR4 expression but the CD40 + IL-4-stimulated cells showed no reduction of chemotactic efficacy. Our finding suggests that changing the SDF-1alpha response of the EBV-infected B cells may serve the viral strategy by directing the infected cells into the extrafollicular areas, rather than retaining them in the lymphoepithelium. PMID- 16476058 TI - Peritoneal macrophages suppress T-cell activation by amino acid catabolism. AB - T-lymphocyte activation triggered by anti-CD3, endogenous or exogenous superantigen, and mitogens was suppressed in a cell-dose-dependent fashion by peritoneal cavity (PerC) leucocytes. Study of lymphocyte-deficient mice and the use of multiparameter fluorescence-activated cell sorter analyses revealed that macrophages were responsible for this form of immune regulation. Interferon-gamma was essential to trigger suppression, which, by enzyme inhibition studies, was shown to be the result of tryptophan and arginine catabolism. These results illustrate that macrophages, which are classically defined by their innate effector function as antigen-presenting cells, have the potential to temper adaptive immunity. PMID- 16476059 TI - CTLA-4 interacts with STAT5 and inhibits STAT5-mediated transcription. AB - Cytotoxic T-lymphocyte antigen-4 (CTLA-4; CD152) is a member of the immunoglobulin gene superfamily with strong homology to the receptor CD28 with which it shares the ligands CD80 and CD86. Unlike CD28, a potent costimulator of T-cell responses, CTLA-4 is transiently expressed on the cell surface of activated T cells and appears to operate predominantly as a negative regulator of T-cell proliferation. Signal transduction mechanisms utilized by CTLA-4 remain unclear although several mechanisms have been implicated. In this study, we show that the cytoplasmic domain of CTLA-4, but not of CD28, binds to STAT5 in yeast two-hybrid assay and in coimmunoprecipitation assays. Mutations of Tyr165 and Tyr182 in CTLA-4 did not abrogate the interaction of STAT5 with CTLA-4. Finally, the overexpression of CTLA-4 in Jurkat T cells inhibits STAT-mediated activation of STAT5 responsive elements. These results suggest that CTLA-4 and STAT5 interact in T cells and that this interaction is important for CTLA-4 signalling. PMID- 16476061 TI - Mice overexpressing p40 in lungs have reduced leucocyte influx and slightly impaired resistance during tuberculosis. AB - Interleukin (IL)-12 (p70) is a heterodimeric cytokine composed of p40 and p35, that plays a major role in the protective immune response to Mycobacterium tuberculosis. To define the role of p40 in lungs during pulmonary M. tuberculosis infection we generated transgenic (Tg) mice overexpressing p40 under control of the surfactant protein C promoter. Tg mice expressed the transgene in their lungs, yet demonstrated elevated pulmonary p40 protein levels. After infection, Tg mice displayed higher pulmonary p40 and p70 levels than wild type mice. Interferon-gamma concentrations were similar in uninfected and infected Tg and wild type mice, arguing against agonistic effects of p40. Tg mice demonstrated reduced recruitment of macrophages, lymphocytes and neutrophils to the lungs early after infection. This was accompanied by reduced pulmonary tumour necrosis factor-alpha, macrophage inflammatory protein (MIP)-2 and MIP-1alpha levels. This suggests that elevated p40 concentrations inhibited the chemotactic effects of p70 on leucocytes. Furthermore, Tg mice displayed slightly higher pulmonary mycobacterial outgrowth late in the infection than wild type mice. Taken together, we demonstrate that constitutive overexpression of p40 in lungs negatively influences IL-12-mediated leucocyte migration and protection against lung tuberculosis. This suggests a novel antagonistic role for p40 homodimers in regulating the chemotactic bioactivity of IL-12 after pulmonary mycobacterial infection. PMID- 16476060 TI - Blockade of OX40-ligand after initial triggering of the T helper 2 response inhibits mercuric chloride-induced autoimmunity. AB - Mercuric chloride (HgCl2)-induced autoimmunity in Brown Norway rats is a spontaneously resolving autoimmune response driven by the activation of T helper type 2 lymphocytes (Th2 cells). Treatment with antibody to OX40-ligand (OX40-L) from the time of the first HgCl2 injection for 12 days had little effect. Delayed treatment commenced 8 days after the first HgCl2 injection significantly suppressed immunoglobulin E production, splenomegaly, weight loss and mortality. This makes OX40/OX40-L signalling an attractive therapeutic target for Th2-driven autoimmune diseases. Intravenous administration of the murine antibody to OX-40-L elicited a vigorous anti-mouse immunoglobulin antibody response that was significantly enhanced compared to the response to control immunoglobulin. It is likely that this response significantly reduced the plasma half-life of the anti OX40-L antibody and this observation has clear implications for the interpretation of data from experiments where anti-OX40-L is used in vivo. PMID- 16476066 TI - The molecular and clinical impact of hepatocyte growth factor, its receptor, activators, and inhibitors in wound healing. AB - Wound healing involves a number of cellular and molecular events, many of which are controlled by soluble growth factors. In the process of healing, hepatocyte growth factor, a cytokine known to act as mitogen, motogen, and morphogen, has been postulated to play multiple roles during several stages of this complex biological process. Produced primarily by stromal fibroblasts, hepatocyte growth factor regulates angiogenesis, vascular permeability, cell migration, matrix deposition and degradation, and other biological processes. The current article discusses recent progress in understanding the multiple roles played by this growth factor in tissue repair. PMID- 16476062 TI - Enhancement of antitumour immunity by a novel chemotactic antigen DNA vaccine encoding chemokines and multiepitopes of prostate-tumour-associated antigens. AB - DNA vaccines provide an attractive technology against cancer because of their safety record in humans and ease of construction, testing and manufacture. In this study, several DNA fragments encoding multiple cytotoxic T lymphocyte (CTL) and T helper cell epitopes were selected from human prostate-specific membrane antigen (hPSM), mouse prostatic acid phosphatase (mPAP), and human prostate specific antigen (hPSA). These DNA fragments were ligated together to form a novel fusion gene, termed the 3P gene. The secondary lymphoid tissue chemokine (SLC), 3P and human immunoglobulin G Fc genes were inserted into pcDNA3.1 to construct a DNA vaccine, designated pSLC-3P-Fc. After vaccination, the DNA is taken up by cells that produce and secrete the SLC-3P-Fc fusion proteins, termed chemotactic antigen (chemo-antigen). The secreted chemo-antigens, in addition to promoting the co-localization of naive, non-polarized memory T cells and dendritic cells, are efficiently captured and processed by dendritic cells via receptor-mediated endocytosis and then cross-presented to both major histocompatibility complex class I and class II in a cognate manner. The results of this study demonstrate that vaccination with pSLC-3P-Fc by gene gun inoculation induced a strong antitumour response in a mouse tumour model, which significantly inhibited tumour growth and prolonged the survival time of the tumour-bearing mice. In vitro, the secreted SLC-3P-Fc fusion protein can attract lymphocytes from human peripheral blood mononuclear cells (PBMC); when human lymphocytes were stimulated by pSLC-3P-Fc-transfected autologous PBMC, CTLs were induced which could specifically kill hPSM-, hPAP-, or hPSA-expressing tumour cells. These observations provide a new vaccine strategy for cancer therapy through promoting the co-localization of lymphocytes and the concomitant enhancement of antigen-specific CD4+ helper and CD8+ cytotoxic T-cell responses against tumour. PMID- 16476067 TI - Risk of amputation in patients with diabetic foot ulcers: a claims-based study. AB - The objective of this study was to undertake a retrospective analysis of claims data of diabetic foot ulcer (DFU) patients to determine the rates of amputation and identify the risk and protective factors. Rates of amputation were calculated in patients diagnosed with DFU in the MEDSTAT Marketscan database between January 2000 and December 2002, who had prediagnosis coverage of 90 days. A nested case control study was conducted using a 1 : 10 ratio of amputee cases to randomly selected nonamputee controls matched on follow-up days. The association of co morbid conditions, demographic factors, and severity (5+ outpatient claims for DFU) on amputation was estimated by adjusted odds ratios (AOR) with 95% confidence intervals (CIs). The 5911 eligible patients yielded an incidence density rate of 2.30 amputations per 100 person years (95% CI = 1.91, 2.77). The 116 cases and 1153 controls averaged 307.3 and 308.5 observation days, respectively. Amputation was significantly increased by male gender (AOR 1.98), Charlson co-morbidity scores of 4-5 and 6+ (AOR = 2.89 and 5.36, respectively), renal disease (AOR = 2.11), peripheral vascular disease (AOR = 2.67), and 5+ outpatient DFU services (AOR = 2.17). Practitioners may consider more aggressive care and earlier referral to specialists for DFU patients who fit risk profiles for amputation, which include peripheral vascular disease, multiple co-morbid conditions, and repeated outpatient DFU services. PMID- 16476068 TI - Intralesional cryosurgery enhances the involution of recalcitrant auricular keloids: a new clinical approach supported by experimental studies. AB - To explain the mechanism of action of a novel intralesional cryoprobe, thermal behavior measurements, and histological studies were performed in swine muscle specimens after intralesional cryosurgery ex vivo. Slow cooling (20 degrees C/min) and thawing (25 degrees C/min) rates, end temperature of -30 degrees C, produced 8 mm wide diffuse coagulative-type necrosis and a 3 mm-wide transition zone around the cryoprobe. In contrast, contact cryosurgery showed fast cooling and thawing rates (80 degrees C/min) and an end temperature of -100 degrees C. Efficacy and safety of the intralesional cryoprobe was further assessed in ten recalcitrant auricular keloids in nine Caucasian patients. There was a 67.4 +/- 23 percent reduction of scar volume at the end of the 18-month follow-up period after a single intralesional treatment (p < 0.005). Significant reduction of hardness, elevation, and redness as well as itching, pain, and tenderness was documented. The histomorphometric analysis, including spectral and fractal analysis, as well as assessment of the fast Fourier transform algorithm, showed parallel alignment and reorganization of the collagen fibers in the treated scar similar to that in the normal dermis. A long hold time in the deep scar core caused minimal damage to the superficial tissue, including melanocytes. There was no evidence of permanent hypopigmentation, active bleeding, infection, or recurrence. The major advantages of the intralesional cryoprobe, including the marked efficacy of a single treatment, may have a major importance in the clinical application of cryosurgery in the treatment of keloids and of other lesions localized deep in the skin. PMID- 16476069 TI - Mitogenic bovine whey extract modulates matrix metalloproteinase-2, -9, and tissue inhibitor of matrix metalloproteinase-2 levels in chronic leg ulcers. AB - Matrix metalloproteinases (MMPs) and their tissue inhibitors play important roles in the wound-healing process. An imbalance in the expression of these molecules is thought to contribute to the failure of chronic ulcers to heal. We investigated whether a mitogenic bovine whey extract enriched with growth factors modulated the expression and activity of MMP-2 and -9, and the tissue inhibitor of MMP-2 (TIMP-2) in chronic leg ulcers. Wound fluids and biopsies were collected from chronic leg ulcer patients whose ulcers were treated topically for 4 weeks with placebo or mitogenic bovine whey extract at concentrations of 2.5, 10, and 20 mg/mL. The levels of MMP-2 and -9 in wound fluid samples was assessed by gelatin zymography and showed a decrease in active MMP-2 in the 2.5 and 10.0 mg/mL mitogenic bovine whey extract-treated ulcers compared with placebo (p<0.05). Immunohistochemical analysis of ulcer biopsies for MMP-2, -9, and TIMP 2 expression showed a reduction in the number of MMP-2-positive dermal fibroblasts in the mitogenic bovine whey extract-treated ulcers compared with pretreatment biopsies (p<0.05) that persisted over the course of the study. In contrast, a transient increase in the number of MMP-9- and TIMP-2-positive cells was observed in mitogenic bovine whey extract treated ulcer biopsies compared with pretreatment levels (p<0.05). These results show that topical application of mitogenic bovine whey extract was able to modulate the expression of MMP-2, -9, and TIMP-2 in chronic leg ulcers and that its constituent growth factors may have the potential to redress the proteolytic imbalance observed in nonhealing chronic ulcers. PMID- 16476070 TI - Cytokeratin, filaggrin, and p63 expression in reepithelialization during human cutaneous wound healing. AB - Cytokeratin (CK), filaggrin (filament aggregating protein), and p63 expression and cellular distribution during reepithelialization has not been systemically studied in the healing stage of human cutaneous wounds. We examined these proteins by immunohistochemical methods in 12 cases of skin ulcer, using seven anti-keratin antibodies, anti-filaggrin, and anti-p63 antibody. At the edge of the wound in skin ulcers, CK1 and 10 expression was reduced, while CK14, 16, and 17 expression was raised. Beneath the wound bed, all layers of the epidermal tongue, deriving from sweat ducts, were positive for CK14 and 17. Both cytokeratins were also found in basal and luminal cells of the dermal duct. CK expression by epithelia continuous with hair follicles showed that, CK14, 16, and 17 were present, and CK1 and 10 were absent. Filaggrin expression was elevated in reepithelialized epithelium. Expression of p63 expression was verified in the suprabasal layer in reepithelialized epithelia. CK, filaggrin, and p63 expression in the reepithelialization stage at the wound edge and at epidermal appendages remaining in the wound bed is undifferentiated and hyperproliferative. The presence of CK14 and 17 in the remaining epidermal appendages in the pathological wound may be important in epidermal replacement. PMID- 16476071 TI - Genetic analysis of skin wound healing and scarring in a porcine model. AB - Contraction is a normal part of skin wound healing and wound closure; however, excessive contraction and severe scarring concern patients and physicians alike. The present study has investigated the degree and kinetics of wound contraction in a porcine model of wound healing, to elucidate the genetic and molecular basis for abnormal skin wound healing and scarring. Healing of excisional skin wounds in juvenile female Yorkshire pigs closely resembled normal healing in humans. In contrast, identical wounds in female red Duroc pigs contracted significantly more, forming hypercontracted, hyperpigmented scars. Yorkshire x red Duroc F1 animals healed without hyperpigmentation, but with significantly greater wound contraction than observed in either parent breed. To examine the genetic transmission of the hypercontractile phenotype, all F1 animals were bred to a single Yorkshire boar, generating 20 backcross animals. All backcross animals healed with significantly less contraction than the normal Yorkshire animals. These findings suggest that the genetic contribution to scar phenotype in this animal model is complex, with a limited number of major genes controlling wound contraction, and an unknown number of minor genes that appear to modulate the impact of the major genes. PMID- 16476072 TI - Perflubron emulsion increases subcutaneous tissue oxygen tension in rats. AB - Oxygen plays a central role in wound healing. The hypothesis of this study was that the administration of a perfluorocarbon emulsion, which dissolves oxygen will increase subcutaneous tissue oxygen tension (PsqO2) in normovolemic and hemorrhaged animals. In the first set of experiments, PsqO2 was measured with a polarographic oxygen electrode along the dorsum of normovolemic-anesthetized rats (n=20) breathing supplemental oxygen. After baseline equilibration of tissue oxygen tension, perflubron emulsion (n=12) or saline control (n=8) was administered. Perflubron administration increased PsqO2 by 32.0 +/- 7.2 mmHg, whereas saline administration had no effect (-1.0 +/- 6.6 mmHg). In a second set of experiments, PsqO2 (n=12) was measured in rats breathing 100 percent oxygen after being bled 20 percent of their blood volume. Creating hypovolemic rats allowed for the study of the effect of perflubron emulsion administration on subcutaneous oxygen tension in tissues with compromised blood flow. Perflubron emulsion was only effective at increasing oxygen tension in tissues with oxygen extractions less then 2.0 vol%. These findings agree with those results analytically predicted based on the oxygen solubility of the perflubron emulsion. PMID- 16476073 TI - A Rho-dependent actin purse-string is involved in wound repair in the early chick amnion following surgical puncture. AB - This study examined the mechanism of wound repair in the early chick amnion following surgical puncture. The chick amnion is a bilayered membrane with ultrastructural features similar to the human amnion, and thus may provide a model of the consequences of amnion puncture following first trimester amniocentesis in humans. Chick amnion was wounded on day 4-5 of incubation. The rate of wounding was measured as 40 microm2/minute by hourly measurements of wound area from initial wounding to a point where the wound appeared to be fully closed. Morphological changes were examined by scanning electron microscopy of amniotic membranes with the overlying chorionic membrane removed. Cells bordering the wound arranged themselves circumferentially during the healing period and eventually came together as a cellular pile up as the healing process was complete. The presence of an actin cable was revealed by fluorescein isothiocyanate-labeled phalloidin. The actin was circumferentially arranged around the wound margin and appeared within 10 minutes after wounding. Treatment of the amnion with the inhibitor of endogenous Rho, C3 exotransferase, inhibited actin cable formation, suggesting that formation of an actin cable within the amnion during wound healing is Rho dependent. PMID- 16476074 TI - In vitro migration and adhesion of fibroblasts from different phases of palatal wound healing. AB - Cleft palate patients often show mid-facial growth impairment after surgical closure of the defect. This is a consequence of palatal wound healing, and more specifically of wound contraction and scar tissue formation. Cells of the fibroblast lineage are responsible for these processes and they display different phenotypes in the course of the wound healing process. The aim of this study was to analyze the in vitro adhesion and migration of wound fibroblasts, isolated during the palatal wound healing process in the rat. Additionally, we analyzed the expression of beta1 integrins and vinculin, the key players in adhesion and migration. Palatal fibroblasts from age-matched controls were analyzed to measure the effects of normal aging. Palatal fibroblasts from unwounded tissue showed a low migratory behavior (<25 microm), a strong capability to adhere (>80%) and a low expression of beta1 integrins and vinculin. In contrast, fibroblasts obtained from healing palatal wounds were highly migratory (>200 microm) coupled to a weak capability to adhere (<65%) and a high expression of vinculin and beta1 integrins. These data show that the palatal wound healing process induces a change in fibroblast phenotype from "quiescent" to "activated," which persists in vitro. PMID- 16476075 TI - Human U937 monocyte behavior and protein expression on various formulations of three-dimensional fibrin clots. AB - Fibrin sealant products such as Tisseel (Baxter Healthcare Corporation) are used in hemostasis and tissue sealing. Tisseel consists of two components, a fibrinogen-containing component and thrombin, which when mixed together form a fibrin clot. There is an interest in delivering monocytes to the wound because they are known to play an important role in the wound-healing process. Therefore, we were interested in finding the best fibrin formulation for delivering monocytes by examining monocyte behavior on 3D-fibrin clots. Using standard adhesion and proliferation assays, we found that monocytes differentially adhere, proliferate, and cluster on and within the 3D-fibrin clots depending on the final fibrinogen and thrombin concentration. Moreover, using a Boyden chamber assay, we found that monocytes migrated through the 3D-fibrin clots in 1-2 days. Furthermore, the protein expression in monocytes seeded on 3D-fibrin clots for various time points varied depending on the fibrinogen and thrombin concentrations in the final 3D-fibrin clots. The above data suggest that various formulations of fibrin sealant Tisseel present a good surface for monocyte adhesion, proliferation and migration in vitro and potentially during the wound healing process in vivo. Moreover, because the monocytes proliferated well and clustered in the 3D fibrin, Tisseel could be a good delivery vehicle for delivering monocytes into chronic wounds to overcome a healing deficiency. PMID- 16476076 TI - Skin wounds in the MRL/MPJ mouse heal with scar. AB - Adult MRL/MpJ mice regenerate cartilage during repair of through-and-through ear punch wounds. However, the ability of this mouse strain to heal isolated cutaneous wounds by regeneration or with scar is unknown. The purpose of this study was to characterize the rate of reepithelialization and collagen architecture in dermal wounds from MRL/MpJ mice compared with C57bl/6 and Balb/c strains. Full-thickness incisional (5 mm) and excisional (2 mm diameter) skin wounds were made on the dorsum of 7-week-old MRL/MpJ, C57bl/6, and Balb/c mice. Ear punch wounds were made simultaneously on each animal. Reepithelialization was complete by 48 hours for incisional skin wounds in each strain. All excisional wounds showed incomplete reepithelialization at 24, 48, and 72 hours. At 14 days, all skin wounds had grossly healed. In contrast to the ear wounds made in C57bl/6 and Balb/c mice, MRL/MpJ ear wounds were completely healed by day 28. Dorsal skin wound sections at 14 and 28 days revealed dense collagen deposition and similar degrees of fibrosis between the three strains of mice. In conclusion, in contrast to wound healing in the ear, MRL/MpJ mouse dorsal cutaneous wounds heal similarly to C57bl/6 and Balb/c mice with dermal collagen deposition and scar formation. PMID- 16476077 TI - Mutual dependence of murine fetal cutaneous regeneration and peripheral nerve regeneration. AB - Mammalian fetal cutaneous wounds made at certain developmental stages show complete regeneration. It is reported that wound healing in both adult and fetal skin is disrupted by denervation. Furthermore, fetal cutaneous regeneration has unique aspects such as epidermal wrinkle texture regeneration and dermal regeneration that depend on developmental stage. Therefore, we have examined the relationship of fetal cutaneous regeneration with denervation. We made cutaneous wounds on fetal mice at various developmental time points including embryonic days (E)13, E15, and E17, and compared the regenerating patterns of peripheral nerves in the skin. We found that when the fetuses are wounded at an early stage of development, peripheral nerves regenerate quicker than at later stages of development when peripheral nerve regeneration is delayed. Next, we denervated the intercostal nerves and made wounds at the denervated sites on E13 and E15. We found that epidermal wrinkling and dermal regeneration were disrupted by denervation. These findings indicate that components of fetal cutaneous regeneration and peripheral nerve regeneration are mutually dependent. PMID- 16476078 TI - Inflammatory signalling pathways involved in astroglial activation by unconjugated bilirubin. AB - During neonatal hyperbilirubinaemia, astrocytes activated by unconjugated bilirubin (UCB) may contribute to brain toxicity through the production of cytokines. As a first step in addressing the signal transduction cascades involved in the UCB-induced astroglial immunological response, we tested whether tumour necrosis factor (TNF)-alpha receptor 1 (TNFR1), mitogen-activated protein kinase (MAPK) and nuclear factor kappaB (NF-kappaB) would be activated in astrocytes exposed to UCB, and examined the profile of cytokine production. Astrocyte cultures stimulated with UCB showed a rapid rise in TNFR1 protein levels, followed by activation of the MAPKs p38, Jun N-terminal kinase1/2 and extracellular signal-regulated kinase1/2, and NF-kappaB. Interestingly, the induction of these signal effectors preceded the early up-regulation of TNF-alpha and interleukin (IL)-1beta mRNAs, and later secretion of TNF-alpha, IL-1beta and IL-6. Treatment of astrocytes with UCB also induced cell death, with levels comparable to those obtained after exposure of astrocytes to recombinant TNF alpha and IL-1beta. Moreover, loss of cell viability and cytokine secretion were reduced when the NF-kappaB signal transduction pathway was inhibited, suggesting a key role for NF-kappaB in the astroglial response to UCB. These results demonstrate the complexity of the molecular mechanisms involved in cell injury by UCB during hyperbilirubinaemia and provide a basis for the development of novel therapeutic strategies. PMID- 16476079 TI - Effects of congenital stationary night blindness type 2 mutations R508Q and L1364H on Cav1.4 L-type Ca2+ channel function and expression. AB - At least 48 mutations in the CACNA1F gene encoding retinal Ca(v)1.4 L-type Ca(2+) channels have been linked to X-linked recessive congenital stationary night blindness type 2 (CSNB2). A large number of these are missense mutations encoding full-length alpha1-subunits that can potentially form functional channels. We have previously shown that such missense mutations can confer their phenotype by different pathological mechanisms, such as complete lack of alpha1 subunit protein expression or dramatic changes in channel gating. Here we investigated the functional consequences of CSNB2 missense mutations R508Q and L1364H. We found no (R508Q) or only minor (L1364H) changes in the gating properties of both mutants after heterologous expression in Xenopus laevis oocytes (at 20 degrees C). However, both mutants resulted in altered expression density of Ca(v)1.4 currents. When expressed in the mammalian cell line tsA-201, the current amplitude of L1364H channels was reduced when cells were grown at 30 degrees C and both mutations affected total alpha1 protein expression. This effect was temperature dependent. Our data provide evidence that, in contrast to previously characterized CSNB2 missense mutations, the clinical phenotype of R508Q and L1364H is unlikely to be explained by changes in channel gating. Instead, these mutations affect the protein expression of Ca(v)1.4 Ca(2+) channels. PMID- 16476080 TI - Is endogenous D-serine in the rostral anterior cingulate cortex necessary for pain-related negative affect? AB - Functional activation of NMDA receptors requires co-activation of glutamate- and glycine-binding sites. D-serine is considered to be an endogenous ligand for the glycine site of NMDA receptors. Using a combination of a rat formalin-induced conditioned place avoidance (F-CPA) behavioral model and whole-cell patch-clamp recording in rostral anterior cingulate cortex (rACC) slices, we examined the effects of d-amino acid oxidase (DAAO), an endogenous D-serine-degrading enzyme, and 7-chlorokynurenate (7Cl-KYNA), an antagonist of the glycine site of NMDA receptors, on pain-related aversion. Degradation of endogenous D-serine with DAAO, or selective blockade of the glycine site of NMDA receptors by 7Cl-KYNA, effectively inhibited NMDA-evoked currents in rACC slices. Intra-rACC injection of DAAO (0.1 U) and 7Cl-KYNA (2 and 0.2 mM, 0.6 microL per side) 20 min before F CPA conditioning greatly attenuated F-CPA scores, but did not affect formalin induced acute nociceptive behaviors and electric foot shock-induced conditioned place avoidance. This study reveals for the first time that endogenous D-serine plays a critical role in pain-related aversion by activating the glycine site of NMDA receptors in the rACC. Furthermore, these results extend our hypothesis that activation of NMDA receptors in the rACC is necessary for the acquisition of specific pain-related negative emotion. Thus a new and promising strategy for the prevention of chronic pain-induced emotional disturbance might be raised. PMID- 16476081 TI - Axonal regeneration in adult CNS neurons--signaling molecules and pathways. AB - Failure of severed adult CNS axons to regenerate could be attributed to both a reduced intrinsic capacity to grow and an heightened susceptibility to inhibitory factors of the CNS extracellular environment. A particularly interesting and useful paradigm for investigating CNS axonal regeneration is its enhancement at the CNS branch of dorsal root ganglion (DRG) neurons after conditional lesioning of their peripheral branch. Recent reports have implicated the involvement of two well-known signaling pathways utilizing separate transcription factors; the Cyclic AMP (cAMP) response element binding protein (CREB) and signal transducer and activator of transcription 3 (STAT3), in conditional lesioning. The former appears to be the pathway activated by neurotrophic factors and Bcl-2, while the latter is responsible for the neurogenic effect of cytokines [such as the leukemia inhibitory factor (LIF) and interleukin-6 (IL-6) elevated at lesion sites]. Recent findings also augmented earlier notions that modulations of the activity of another class of cellular signaling intermediate, the conventional protein kinase C (PKC), could result in a contrasting growth response by CNS neurons to myelin-associated inhibitors. We discuss these signaling pathways and mechanisms, in conjunction with other recent reports of regeneration enhancement and also within the context of what is known about aiding regeneration of injured CNS axons. PMID- 16476082 TI - Autoradiographical distribution of imidazoline binding sites in monoamine oxidase A deficient mice. AB - This study has used receptor autoradiography to characterize imidazoline binding sites (I-BS) in monoamine oxidase (MAO) A knockout and wild-type mice. A comparison between MAO-A and MAO-B, binding of the endogenous beta-carboline [(3)H]harmane, and I-BS, has been made using sections from brain and kidney. The loss of binding to MAO-A in the knockout animals was confirmed using the selective radioligand [(3)H]Ro41-1049, with labelling reduced to background levels. The binding of [(3)H]Ro19-6327 to MAO-B was unaffected, indicating no change in this isoform in response to the loss of MAO-A. A reduction in binding to the I(2)-BS, as labelled by both [(3)H]idazoxan and [(3)H]2-BFI (2-(2 benzofuranyl)-2-imidazoline), was seen in the MAO-A knockout animals in both brain and kidney sections, whereas binding to the I(1)-BS in kidney sections remained unchanged. The loss of I(2) binding was found to be regionally dependent and was positively correlated with the relative expression of MAO-A in specific regions in the wild-type animals. Using the MAO-A knockout mice it was also possible to demonstrate a non-MAO-A population of binding sites labelled by the putative I-BS endogenous ligand, harmane. PMID- 16476084 TI - Current issues facing coagulationists--meeting report. PMID- 16476083 TI - Stromal cell-derived factor-1alpha modulation of the excitability of rat substantia nigra dopaminergic neurones: presynaptic mechanisms. AB - In rat substantia nigra (SN), Chemokine (CXC motif) receptor 4 (CXCR4) for the chemokine stromal cell-derived factor (SDF)-1alpha is expressed on dopaminergic (DA) neurones, but also on non-DA cells, suggesting presynaptic actions. Using whole-cell patch-clamp recordings in DA neurones of rat SN slices at a holding potential of -60 mV, we showed here that SDF-1alpha exerts multiple presynaptic effects. First, SDF-1alpha (10 nm) induced an increase in the frequency of spontaneous and miniature GABA(A) postsynaptic currents by presynaptic mechanisms, consistent with the presence of CXCR4 on GABAergic neurones of the SN, as revealed by immunocytochemistry. Second, SDF-1alpha (0.1-1 nm) induced a glutamatergic inward current resistant to tetrodotoxin (TTX), most probably the result of glutamate release from non-neuronal cells. This inward current was not blocked by the CXCR4 antagonist AMD 3100 (1 microm), consistent with the lack of CXCR4 on astrocytes as shown by immunocytochemistry under basal conditions. Finally, SDF-1alpha (10 nm) induced, via CXCR4, an outward G protein-activated inward rectifier (GIRK) current, which was TTX sensitive and prevented by application of the GABA(B) antagonist CGP55845A, suggesting GABA spillover on to GABA(B) receptors. Our results show that SDF-1alpha induces, via presynaptic mechanisms, alterations in the excitability of DA neurones as confirmed by current-clamp experiments. PMID- 16476085 TI - Current co-ordinated activities of the PEDNET (European Paediatric Network for Haemophilia Management). PMID- 16476086 TI - Occurrence of inhibitors in previously untreated or minimally treated patients with haemophilia A after exposure to a plasma-derived solvent-detergent factor VIII concentrate. AB - A solvent-detergent virus-inactivated plasma-derived FVIII concentrate (SD pdFVIII) has been employed for treatment of Italian patients with haemophilia A for 15 years. This product is a non-monoclonally purified, high purity FVIII concentrate, containing large amounts of von Willebrand factor (VWF). A retrospective survey was carried out in Italy in order to evaluate the immunogenicity of SD-pdFVIII in previously untreated patients (PUPs) or in minimally treated patients (MTPs), i.e. previously exposed for up to 5 days only to other plasma-derived concentrates. The survey included 99 patients with ages ranging from 6 to 64 years (median=21.3) of whom 31 PUPs and 68 MTPs, the latter with a median of four exposure days (EDs; range 1-5) to other plasma products. Surveyed patients had been exposed to SD-pdFVIII for a median of 83 EDs (range 21 1580). Seven patients (three PUPs and four MTPs), all with severe haemophilia, had developed inhibitors [7.1%, 95%; confidence interval: 3-14%] after a median of 11 EDs (range 4-22). Of them, two were low responders ( or = 80 years, compared with those with fathers whose age at death was <60 years. A similar reduction was found when the age at death of mothers was > or = 85 years compared with <65 years. Furthermore, the risk reduction was more evident amongst persons with both parents being long-lived parents compared with those with being short-lived parents, especially for death from cardiovascular disease. CONCLUSIONS: Our findings indicate that parental longevity could be a predictor for reduced risk of mortality from stroke, cardiovascular disease, and all causes for both Japanese men and women. PMID- 16476107 TI - Gains and losses of warfarin therapy as performed in an anticoagulation clinic. AB - BACKGROUND: Knowledge of the net benefit of warfarin therapy in routine care is needed to define realistic management recommendations, but lack of randomized controls precludes conventional risk-benefit analysis. OBJECTIVE: Assess risk and benefit of routine warfarin therapy in an anticoagulation clinic. DESIGN: Retrospective observational analysis. PATIENTS: A total of 1435 outpatients on warfarin for a total of 1613 patient years, treated to prevent the target events recurrent venous thromboembolism (VTE) or myocardial infarction (MI), and stroke in patients with atrial fibrillation (AF) or mechanical heart valves. MEASUREMENTS: Major bleeding and thromboembolic (TE) events and all deaths. CALCULATIONS: Expected annual target event rates without warfarin were from published data. Differences between combined major events observed with warfarin, and expected without warfarin were calculated. RESULTS: In the total material, annual rates were 3.0% major TE events, 1.1% major bleeding events, 0.12% fatal bleeding, and a benefit/risk ratio of 3.8. The net gain, expressed in reduced combined bleeding and target TE annual event rate, was 9.9% in secondary prophylaxis in AF, 4.4% in VTE patients, 2.7% in post-MI patients, 2.4% in primary prophylaxis in AF and 0.6 in patients with mechanical heart valves. The apparent benefit/risk ratio was 3.9 in VTE patients, 5.8 in AF patients and 1.1 in patients with mechanical heart valves. CONCLUSION: Net effects of prolonged warfarin therapy in patients with VTE and AF performed in an anticoagulation clinic have an acceptable risk/benefit ratio, comparable with what has been obtained in elective clinical trials. PMID- 16476108 TI - Fatal nonocclusive mesenteric ischaemia: population-based incidence and risk factors. AB - OBJECTIVES: To estimate the incidence and extension of visceral organ infarction, and to evaluate potential causes, in patients with autopsy-verified nonocclusive mesenteric ischaemia (NOMI) and transmural intestinal infarction. SETTING: In Malmo, Sweden, the autopsy rate between 1970 and 1982 was 87%, creating possibilities for a population-based study. DESIGN: Amongst 23 446 clinical autopsies, 997 cases were coded for intestinal ischaemia in a database. In addition, 7569 forensic autopsy protocols were analysed. In a nested case-control study within the clinical autopsy cohort, four NOMI-free controls, matched for gender, age at death and year of death, were identified for each fatal NOMI case to evaluate risk factors. RESULTS: The overall incidence of autopsy-verified fatal NOMI was 2.0/100,000 person-years, increasing with age up to 40/100,000 person-years in octogenarians. Patients with stenosis of the superior mesenteric artery (SMA; n = 25) were older (P = 0.002) than those without (n = 37), and had more often a concomitant stenosis of the coeliac trunk (P < 0.001). Synchronous infarction in the liver, spleen or kidney occurred in one-fifth of all patients. Fatal cardiac failure [OR 2.9 (1.7-5.2)], history of atrial fibrillation [OR 2.2 (1.2-4.0)] and recent surgery [OR 3.4 (1.6-6.9)] were risk factors for fatal NOMI. CONCLUSIONS: Fatal heart failure was the leading cause of intestinal hypoperfusion, although stenosis of the SMA and coeliac trunk, atrial fibrillation and recent surgery contributed significantly. Collaboration across specialties seems to be of utmost importance to improve the prognosis. PMID- 16476109 TI - Obesity and cardiovascular risk factors in type 2 diabetes: results from the Swedish National Diabetes Register. AB - OBJECTIVES: To compare obese with normal and overweight type 2 diabetic patients regarding body mass index (BMI) and cardiovascular risk factors, and to analyse changes in weight versus risk factors. DESIGN AND SETTING: A cross-sectional study of 44 042 type 2 patients, and a 6-year prospective study of 4468 type 2 patients. RESULTS: Obese patients (BMI > or = 30 kg m(-2)), 37% of all patients, had high frequencies of hypertension (88%), hyperlipidaemia (81%) and microalbuminuria (29%). Only 11% had blood pressure <130/80 mmHg. Their ratio of triglycerides to HDL cholesterol was considerably elevated, whilst the mean total and LDL cholesterol were similar as in normal weight subjects. Obese patients had elevated odds ratios for hypertension, hyperlipidaemia and microalbuminuria: 2.1, 1.8 and 1.4 in the cross-sectional study, similarly confirmed in the prospective 6-year study. BMI was an independent predictor of these risk factors (P < 0.001), although only slightly associated with HbA1c and not with total or LDL cholesterol. A change in BMI during the prospective study was related to a change in HbA1c in patients treated with diet and oral hypoglycaemic agents (OHAs) but not with insulin. In all patients, an increase in BMI was related to the development of hypertension, and a change in BMI to change in blood pressure, also mostly confirmed when treated with diet, OHAs or insulin. CONCLUSIONS: The high frequencies of risk factors in obese type 2 patients implies an increased risk of cardiovascular disease and the need for therapeutic measures. The paradox that hypoglycaemic treatment accompanied by weight gain may increase cardiovascular risk factors seems to be verified here concerning hypertension but not concerning microalbuminuria. PMID- 16476110 TI - Airway inflammation in subjects with gastro-oesophageal reflux and gastro oesophageal reflux-related asthma. AB - STUDY OBJECTIVES: Asthma and gastro-oesophageal reflux (GER) are both characterized by airway inflammation. DESIGN: The purposes of this work were (i) to study airway inflammation in patients troubled by gastro-oesophageal reflux (GER) and GER associated with asthma, (ii) to ascertain whether GER can aggravate asthma by exacerbating the pre-existing airway inflammation and oxidative stress and (iii) to establish the validity of analysing breath condensate and induced sputum when studying the airways of subjects affected by GER. PATIENT S AND METHODS: We enrolled 14 patients affected by mild asthma associated with GER (40 +/-12 years), nine with mild but persistent asthma (39 +/- 13 years), eight with GER (35 +/- 11 years) and 17 healthy subjects (37 +/- 9 years). Sputum cell counts and concentrations of interleukin-4 (IL-4), IL-6 and 8-isoprostane were measured in breath condensate and supernatant. MEASUREMENTS AND RESULTS: GER related asthma is characterized by an eosinophilic inflammation, as determined by elevated concentrations of IL-4 in breath condensate and sputum supernatant, and by sputum cell analysis. GER alone presents a neutrophilic pattern of inflammation when determined by elevated concentrations of IL-6 in sputum cell analysis. A concomitant increase has been found in 8-isoprostane in GER associated (or not associated) with asthma. CONCLUSIONS: We conclude that GER is characterized by a neutrophilic airway inflammation and by increased oxidative stress. GER does not however aggravate pre-existing airway inflammation in asthma patients. Determinations of inflammatory and oxidant markers in the breath condensate of subjects with GER reflect these measured in the induced sputum. PMID- 16476111 TI - Tobacco smoke, systemic inflammation and the risk of type 2 diabetes. PMID- 16476114 TI - The efficacy of folk medicines in the management of type 2 diabetes mellitus: results of a randomized controlled trial of Syzygium cumini (L.) Skeels. AB - OBJECTIVE: To investigate whether a tea prepared from leaves of jambolan, Syzygium cumini (L.) Skeels, has an antihyperglycaemic effect in patients with type 2 diabetes mellitus. RESEARCH DESIGN AND METHODS: Patients with type 2 diabetes mellitus were enrolled in a double-blind, double-dummy, randomized clinical trial. The three experimental groups received a tea prepared from leaves of S. cumini plus placebo tablets, placebo tea plus glyburide tablets or placebo tea plus placebo tablets. RESULTS: In total, 27 patients were allocated to one of the treatment groups and followed for 28 days. Fasting blood glucose levels decreased significantly with glyburide and did not change with S. cumini tea or placebo. Body mass index, creatinine, gamma-glutamyl transferase, alkaline phosphatase, aspartate aminotransferase (SGOT), alanine aminotransferase (SGPT), 24-h glicosuria, 24-h proteinuria, triglycerides, total, low-density lipoprotein and high-density lipoprotein cholesterol did not vary significantly between the different groups. CONCLUSIONS: Tea prepared from leaves of S. cumini has no hypoglycaemic effect. PMID- 16476115 TI - Medicines information and adherence in HIV/AIDS patients. AB - BACKGROUND: Providing written medicines information is being legislated in an increasing number of countries worldwide, with the patient information leaflet (PIL) being the most widely used method for conveying health information. The impact of providing such information on adherence to therapy is reportedly unpredictable. Therapy for human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) and related opportunistic infections usually involves polytherapy and complex regimens, both of which are risk factors for non adherence. The objective of this study was to assess the impact of medicines information on adherence to chronic co-trimoxazole therapy in low-literate HIV/AIDS patients. METHODS: Two different PILs were designed for co-trimoxazole tablets and were available in both English and isiXhosa. Participants were randomly allocated to a control group (receiving no PIL), group A (receiving a "complex PIL") and group B (receiving a "simple PIL" incorporating pictograms). At the first interview, demographic data were collected and the time, date and day that the participant would take his/her first tablet of the month's course was also documented. In a follow-up interview adherence to therapy was assessed using two methods; self-report and tablet count. RESULTS: The medicines information materials incorporating simple text and pictograms resulted in significantly improved adherence to therapy in the short term, whereas a non significant increase in adherence was associated with the availability of the more complex information. This was shown by both the self-reported assessment as well as the tablet count. CONCLUSION: This research suggests that appropriately designed written material can have a positive impact in improving adherence and, together with verbal consultation, are essential for enabling patients to make appropriate decisions about their medicine taking. PMID- 16476116 TI - Relationships of tacrolimus pharmacokinetic measures and adverse outcomes in stable adult liver transplant recipients. AB - BACKGROUND AND OBJECTIVES: Alternative measures to trough concentrations [non trough concentrations and limited area under the concentration-time curve (AUC)] have been shown to better predict tacrolimus AUC. The aim of this study was to determine if these are also better predictors of adverse outcomes in long term liver transplant recipients. METHODS: The associations between tacrolimus trough concentrations (C(0)), non-trough concentrations (C(1), C(2), C(4), C(6/8)), and AUC(0-12) and the occurrence of hypertension, hyperkalaemia, hyperglycaemia and nephrotoxicity were assessed in 34 clinically stable liver transplant patients. RESULTS AND DISCUSSION: The most common adverse outcome was hypertension, prevalence of 36%. Hyperkalaemia and hyperglycaemia had a prevalence of 21% and 13%, respectively. A sequential population pharmacokinetic/pharmacodynamic approach was implemented. No significant association between predicted C(0), C(1), C(2), C(4), C(6/8) or AUC(0-12) and adverse effects could be found. Tacrolimus concentrations and AUC measures were in the same range in patients with and without adverse effects. CONCLUSIONS: Measures reported to provide benefit, preventing graft rejection and minimizing acute adverse effects in the early post-transplant period, were not able to predict adverse effects in stable adult liver recipients whose trough concentrations were maintained in the notional target range. PMID- 16476117 TI - Comparison of mycophenolic acid pharmacokinetic parameters in kidney transplant patients within the first 3 months post-transplant. AB - OBJECTIVE: The aim of this study was to investigate the effect of time on pharmacokinetic (PK) parameters of mycophenolic acid (MPA) in the early post transplant period in kidney recipients. MPA is the active metabolite of mycophenolate mofetil (MMF), which was introduced into clinical practice ten years ago. METHOD: Mycophenolate mofetil was co-administered with cyclosporin (CsA) in a subgroup of 23 patients and with tacrolimus (Tac) in a subgroup of 10 patients. MPA plasma concentration profiles were measured by a validated high performance liquid chromatography method 1 week, 2 and 3 months after transplantation. RESULTS: Despite a comparable MMF dose, a large inter-patient variability in both MPA area under the curve (AUC) from 0 to 12 h (range 10.03 135.4 microg h/mL) and in predose concentrations (0.31-6.09 microg/mL) was observed. Patients with AUC > 35 microg h/mL showed better (P < 0.1) renal function than patients with AUC < 20 microg h/mL (mean creatinine concentration 1.48 +/- 0.12 vs. 3.35 +/- 0.4 mg/dL respectively). The total MPA trough and AUC did not correlate with biochemical parameters: leucocyte cell count and haematocrit. A higher trough level of the metabolite MPA glucuronide (MPAG) in the 1 week after transplantation was found when compared with the 3-month level (mean 150.1 +/- 146.7; range 17.1 to 560 vs. 75.8 +/- 40.0; range 27.3 to 174.2 microg/mL). The concentration of MPA, and MPA AUC values were significantly lower in patients receiving MMF and CsA than those receiving MMF and Tac during all three periods studied (P < 0.02). The influence of C(0) and MPA AUC values on the risk of graft rejection was investigated using receiver operating characteristic (ROC) curve analysis. The area under the ROC curve for AUC was 0.847, whereas that of C(0) was 0.632. CONCLUSIONS: The MPA AUC(0-12h) appeared to be the more effective PK parameter for predicting acute rejection. We recommend that routine MPA and MPAG therapeutic drug level monitoring should be an important part of MMF therapy. PMID- 16476118 TI - Screening for sleep disorders in community pharmacies--evaluation of a campaign in Switzerland. AB - BACKGROUND: In 2003 the Swiss federation of pharmacists organized a campaign "sleep disturbances--daytime sleepiness". The goal was to assist pharmacy clients in detecting likely causes of any sleep disturbance or daytime sleepiness through a free of charge screening, and to deliver targeted counselling. For pharmacy practice there are no screening or triage guidelines to assess the severity of sleep and wakefulness disturbances and potential causes for those disturbances. In this paper the outcome of the campaign in terms of feasibility, participation, observed response patterns, sale of over-the-counter (OTC) sleeping pills, and counselling activities is evaluated. METHODS: The Stanford sleep disorders questionnaire and the Epworth sleepiness scale served to identify patterns of symptoms suggestive of four major categories of sleep disorders. The questionnaires were posted on a web-site and the clients' data were entered online in the pharmacies. A report was automatically generated and immediately available online to the pharmacists. The pharmacists documented separately their counselling activities in a pharmacist's activity report. RESULTS: Six hundred and twenty-two (23%) of 2743 pharmacy clients had response patterns suggestive of obstructive sleep apnoea, 418 (15%) of restless-legs-syndrome, 39 (1%) of a sleep disorder potentially associated with a psychiatric condition and 79 (3%) of narcolepsy. An Epworth sleepiness score >10 points was found in 567 (21%). After screening, 2345 (86%) pharmacy clients received targeted counselling. Only 216 (8%) purchased an OTC sleeping pill and 704 (26%) were recommended to consult a physician, but of these, 446 (63%) were already under medical supervision. CONCLUSIONS: The online screening tool for sleep disorders and daytime sleepiness was successfully introduced in Swiss pharmacies. Pharmacies were able to assess the pattern of individual sleep disorders and to identify a possible cause in nearly one-third of the cases. PMID- 16476119 TI - Formulation and characterization of poly (D,L-lactide-co-glycolide) nanoparticle containing vascular endothelial growth factor for gene delivery. AB - OBJECTIVE: The stability, in vitro release, in vitro cell transfection efficiency and in vivo gene transfer of vascular endothelial growth factor (VEGF(165)) plasmid DNA-loaded poly(d,l-lactide-co-glycolide) (PLGA) nanoparticles were investigated. METHODS: DNA-loaded nanoparticles were prepared with PLGA bearing VEGF(165) plasmid DNA and characterized with respect to morphology, size and encapsulation efficiency. The gene transfer efficiency of nanoparticles was tested in vitro on the cultured myocardial cells. And then the suspension of VEGF loaded nanoparticles (VEGF-NPs) was injected into myocardial tissues in vivo to observe the process of nanoparticles as vectors for gene transfer to cardiac myocytes and to detect its biological effect and evaluate angiogenesis. RESULTS: The encapsulation efficiency of the VEGF-NPs was 58.06 +/- 2.8% and their diameter ranged 100-300 nm. VEGF gene could be successfully transfected into myocardial cells by nanoparticles, which significantly enhanced gene transfection efficiency. A great number of nanoparticles were seen in the myocardial cytoplasm and nucleus. Furthermore, the nanoparticles began to dissolve and degrade. There was a significant increase in the number of capillaries in the VEGF-NPs group when compared with the plasmid DNA group. CONCLUSION: The PLGA nanoparticles are capable of DNA delivery to cardiac myocytes for treating ischaemic myocardium. The direct gene transfer of VEGF-NPs into ischaemic rabbit myocardium can improve cardiac function and increase the number of capillaries. PMID- 16476120 TI - Effects of claritromycin on inflammatory parameters and clinical conditions in children with bronchiectasis. AB - BACKGROUND: The effects of the macrolides cannot be ascribed to their antibacterial action alone. Their immunoregulatory and anti-inflammatory functions are significant too. They are frequently used in the treatment of diffuse panbronchiolitis and cystic fibrosis (CF). AIM: To evaluate the effects of a macrolide antibiotic [clarithromycin (CAM)] on the process of inflammation [by measuring IL-8, TNF-alpha, IL-10 levels and cell profiles in bronchoalveolar lavage (BAL) fluid], pulmonary function and sputum production in children with steady-state bronchiectasis, secondary to causes other than CF or primary immunodeficiencies. METHODS: Seventeen patients randomized to the treatment group received CAM and supportive therapies for 3 months and 17 patients in the control group were given supportive therapies only. RESULTS: Compared with the control group, the treatment group showed a significant decrease in IL-8 levels, total cell count, neutrophil ratios in BAL fluid and daily sputum production at the end of the third month. There was also a significant increase in the treatment group's BAL fluid macrophage ratios. The differences in pulmonary function test parameters were not significant. CONCLUSION: Use of CAM in children with steady state bronchiectasis results in laboratory improvement by reducing the inflammatory processes in the lungs. No corresponding clinical improvement could be shown but although this is possible with long-term use, trial validation is necessary. PMID- 16476121 TI - Treatment persistence: a comparison among patients with schizophrenia who were initiated on atypical antipsychotic agents. AB - BACKGROUND: Although clinical trials have demonstrated the efficacy of atypical antipsychotic agents in reducing symptoms of schizophrenia, the likelihood of sustaining control of schizophrenic symptoms may depend on treatment persistence. OBJECTIVE: In this study, we compared treatment persistence between patients who were initiated on risperidone or olanzapine, the two most widely prescribed atypical antipsychotic agents. METHOD: We identified patients with schizophrenia by ICD-9-CM codes (> or =1 inpatient or > or =2 outpatient ICD-9-CM codes > or =7 days apart) between 1 July 1998 and 30 June 1999. We further selected those who were prescribed the target drug during 1 April 1999 through 31 March 2000 provided that they were not on any antipsychotic agents during the prior 6 months. Using event history analysis, we compared the treatment persistence in terms of hazard ratio between olanzapine and risperidone initiators, adjusting for patient's sociodemographic and clinical characteristics. RESULTS: Following the initiation of the target drug, more patients switched from risperidone to olanzapine than vice versa. However, among patients with schizophrenia who had comorbid diabetes, there were more patients who made a switch from olanzapine to risperidone; whereas among those who used anxiolytics, there were more patients who switched from risperidone to olanzapine. Finally, olanzapine initiators had decreased hazards of discontinuation by 14% (unadjusted; P < 0.001) and 12% (adjusted; P = 0.002), respectively, than risperidone initiators. CONCLUSIONS: Compared with risperidone, olanzapine seems to be better tolerated by patients as indicated by better treatment persistence. As such, initiation of olanzapine may increase the likelihood of sustaining control of symptoms of schizophrenia. Future research needs to provide a more comprehensive assessment of treatment persistence by considering other antipsychotic agents in the study and developing models to assess treatment persistence and switching as two interdependent competing risks. PMID- 16476122 TI - The influence of antibiotic use on the occurrence of vancomycin-resistant enterococci. AB - BACKGROUND: Several studies have documented the influence of antibiotic selective pressure, mainly from the use of glycopeptides, third-generation cephalosporins, quinolones and lincosamides, on the frequency of vancomycin-resistant enterococci (VRE) occurrence in hospitals. The aim of this study was to evaluate the relationship between VRE occurrence and antibiotic use in the Department of Hemato-Oncology of the Teaching Hospital in Olomouc (DHO), Czech Republic, over a 6-year period under standard and unchanged hygienic and epidemiological conditions. METHODS: During the period of 1998-2003, Enterococcus sp. strains and VRE were isolated by standard methods from clinical samples taken from DHO in patients. The frequency of VRE occurrence was expressed as the number of isolated strains per 100 bed-days/year. DHO antibiotic consumption data were processed according to the anatomical therapeutic chemical (ATC)/defined daily dose (DDD) system valid in 2003 and expressed in defined daily dose per 100 bed-days (DDD/100 bed-days) for each year of the period. RESULTS: Since 1998, the occurrence of VRE decreased significantly (from 0.28 to 0.17 VRE/100 bed-days in 2001). Between 2001 and 2003, a significant (P < 0.05) increase from 0.17 to 0.38 was observed. The antibiotic use decreased from 205.2 in 1998 to 161.0 DDD/100 bed-days in 1999 and after an increase in 2001 (to 181.8 DDD/100 bed-days) it remained relatively stable. A significant decrease was observed in third generation cephalosporins and quinolones (from 29.5 to 9.7 and from 42.2 to 30.2 DDD/100 bed-days respectively) between 1998 and 1999. In 2002-2003, the use of third-generation cephalosporins and glycopeptides increased substantially (from 10.1 to 13.9 and from 11.3 to 15.2 DDD/100 bed-days respectively). The Pearson correlation value was significantly positive (P < 0.05) for VRE occurrence and the use of glycopeptides and third-generation cephalosporins. CONCLUSIONS: While our study confirms the effect of use of glycopeptides and third-generation cephalosporins on occurrence of VRE, no influence of quinolones and lincosamides over the 6-year period was shown. PMID- 16476123 TI - Trends in the use of oral antidiabetic drugs by outpatients in Taiwan: 1997-2003. AB - OBJECTIVE: To define the prescribing patterns of oral antidiabetic drugs (OADs) in Taiwan over a 7-year period (1997-2003) and to critically comment on changes observed. METHODS: A cross-sectional study design was implemented using data from Taiwan's National Health Insurance Research Database between January 1997 and December 2003. Outpatients who were 18 years or older and had at least an OAD claim during the study period were identified. The unit of analysis was each OAD prescription for diabetic outpatient visits. The prescribing trends were described in terms of annual changes in prescribing rates and patterns. RESULTS: The numbers of OAD prescriptions rose 1.23-fold. The sulfonylurea (SU) class was the most commonly used OAD, but the prescribing rates for this class declined over time. The biguanide (BG) class was the second most frequently prescribed OAD class and its prescribing rate initially increased, peaked in 2000, and then substantially decreased. The largest increase in prescribing was for acarbose use. The prescribing rates of two new classes of OAD, meglitinide (MG) and thiazolidinedione (TZD), also significantly increased within a short period of time. A trend towards combination therapy was observed away from monotherapy. The SU class was the most commonly prescribed as monotherapy. SU plus BG was the most commonly prescribed dual therapy. Triple oral therapy showed a significant ninefold increase. CONCLUSION: The prescribing rates of OADs are shifting from the older OADs (i.e. SUs) to newer OADs [i.e. alpha-glucosidase inhibitor (AGI), MGs, and TZDs]. The prescribing patterns of OADs are moving toward combination therapy, especially triple oral therapy. PMID- 16476124 TI - Inhibition of human cytochrome P450 isoforms and NADPH-CYP reductase in vitro by 15 herbal medicines, including Epimedii herba. AB - OBJECTIVE: We evaluated the potential of 15 herbal medicines (HMs), commonly used in Korea, to inhibit the catalytic activities of several cytochrome P450 (CYP) isoforms and microsomal NADPH-CYP reductase. METHODS: The abilities of 1-1000 microg/mL of freeze-dried aqueous extracts of 15 HMs to inhibit phenacetin O deethylation (CYP1A2), tolbutamide 4-methylhydroxylation (CYP2C9), S-mephenytoin 4'-hydroxylation (CYP2C19), dextromethorphan O-demethylation (CYP2D6), chlorzoxazone 6-hydroxylation (CYP2E1), midazolam 1-hydroxylation (CYP3A4) and NADPH-CYP reductase were tested using human liver microsomes. RESULTS: The HMs Epimedii herba, Glycyrrhizae radix and Leonuri herba inhibited one or more of the CYP isoforms or NADPH-CYP reductase. Of the three HMs, Epimedii herba extracts were the most potent inhibitors of several CYP isoforms (IC(50) 67.5 microg/mL for CYP2C19, 104.8 microg/mL for CYP2E1, 110.9 microg/mL for CYP2C9, 121.9 microg/mL for CYP3A4, 157.8 microg/mL for CYP2D6 and 168.7 microg/mL for CYP1A2) and NADPH-CYP reductase (IC(50) 185.9 microg/mL ). CONCLUSION: These results suggest that some of the HMs used in Korea have the potential to inhibit CYP isoforms in vitro. Although the plasma concentrations of the active constituents of the HMs were not determined, some herbs could cause clinically significant interactions because the usual doses of those individual herbs are several grams of freeze-dried extracts. Controlled trials to test the significance of these results are necessary. PMID- 16476125 TI - Molecular analysis of thiopurine S-methyltransferase alleles in Taiwan aborigines and Taiwanese. AB - BACKGROUND: Thiopurine S-methyltransferase (TPMT) is a cytosolic enzyme involved in the metabolism of these thiopurine drugs. Methylation of thiopurine drugs by TPMT competes with the formation of their active 6-thioguanine nucleotide metabolite, thereby potentially modulating the therapeutic and toxic effects of these drugs. OBJECTIVE: To analyze the thiopurine S-methyltransferase allelic frequencies in Taiwan aborigines and Taiwanese. METHODS: We used polymerase chain reaction-restriction fragment length polymorphism method to determine the allelic frequencies of TPMT variants (TPMT*1-TPMT*8) in 409 Taiwan aborigines and 117 Taiwanese. RESULTS AND DISCUSSION: The results showed that the allelic frequencies of TPMT*1 were 99.88% and 98.72% for Taiwan aborigines and Taiwanese respectively. The allelic frequencies of TPMT*3C were 0.12% and 1.28% for Taiwan aborigines and Taiwanese respectively. No TPMT*2, 3A, 3B, 3D and 4-8 were found in these populations. CONCLUSION: Our results provide useful information for using thiopurine drugs in these populations. PMID- 16476126 TI - The relevance of CYP2D6 genetic polymorphism on chronic metoprolol therapy in cardiovascular patients. AB - BACKGROUND AND OBJECTIVE: CYP2D6 polymorphisms are well described in normal populations but there are few data on its clinical significance. We therefore investigated the influence of CYP2D6 polymorphism on steady-state plasma concentrations and apparent oral clearance of metoprolol in patients with cardiovascular diseases. METHODS: Ninety-one patients on metoprolol were recruited. Plasma concentrations of metoprolol and alpha-hydroxy metoprolol were measured at 4-h post-dose. CYP2D6 genotyping (*1, *3, *4, *5, *9, *8, *10, *17 and duplication) were performed on the DNA extracted. Ratio of plasma concentrations of metoprolol and alpha-hydroxy metoprolol and the apparent oral clearance of metoprolol were calculated. The influences of CYP2D6 genotypes were investigated. RESULTS: A 100-fold variation was noted for both plasma concentrations of metoprolol and alpha-hydroxy metoprolol. There was a weak correlation between the total daily doses and plasma concentrations of both. Plasma concentrations were found to be higher in patients with genotypes that predicted lower enzyme activity. One patient homozygous for CYP2D6*4 had the highest metoprolol concentration per unit dose. With an antimode of 10. Two patients were identified as poor metabolizers (PMs) (2.1%; 95% CI: 2.9). The PMs who had a plasma metabolic ratio (pMR) of 37.8 was homozygous CYP2D6*4 whereas the other with pMR 14.5 was genotyped CYP2D6 *4/*10. There was a 36-fold difference between the highest and lowest clearance values. Large overlaps in the clearance values were noted between most of the genotypes. CONCLUSIONS: Our data suggest that pharmacogenetic measures could be used to design a more individualized metoprolol dosage regimen for patients. PMID- 16476127 TI - Cannabis use and the mental health of young people. AB - OBJECTIVE: To review the evidence on the mental health and psychosocial consequences of rising rates of cannabis use among young people in developed countries. METHOD: This paper critically reviews epidemiological evidence on the following psychosocial consequences of adolescent cannabis use: cannabis dependence; the use of heroin and cocaine; educational underachievement; and psychosis. Leading electronic databases such as PubMed have been searched to identify large-scale longitudinal studies of representative samples of adolescents and young adults conducted in developed societies over the past 20 years. RESULTS: Cannabis is a drug of dependence, the risk of which increases with decreasing age of initiation. Cannabis dependence in young people predicts increased risks of using other illicit drugs, underperforming in school, and reporting psychotic symptoms. Uncertainty remains about which of these relationships are causal although the evidence is growing that cannabis is a contributory cause of psychotic symptoms. CONCLUSIONS: We face major challenges in communicating with young people about the most probable risks of cannabis use (dependence, educational underachievement and psychosis) given uncertainties about these risks and polarized community views about the policies that should be adopted to reduce them. PMID- 16476129 TI - Prevalence and long-term course of lifetime eating disorders in an adult Australian twin cohort. AB - OBJECTIVE: Few studies exist that have examined the spectrum and natural long term course of eating disturbance in the community. We examine the lifetime prevalence and long-term course of anorexia nervosa (AN), bulimia nervosa (BN) and eating disorder not otherwise specified (EDNOS) in an adult female twin population. METHOD: Female twins (n = 1002) from the Australian Twin Registry, aged 28-39 years, were assessed using the Eating Disorder Examination, revised to yield lifetime diagnostic information. For women with lifetime eating disorders, the assessment occurred, on average, 14.52 years (SD = 5.65) after onset of their disorder. RESULTS: In accordance with other community studies, we found a 1.9% lifetime prevalence of AN, with an additional 2.4% who met the criteria for 'partial AN' (absence of amenorrhea). Criteria for BN were met by 2.9% of the women, an additional 2.9% of women met criteria for binge eating disorder, while 5.3% met criteria for purging disorder unaccompanied by binge eating (EDNOS-p). Eleven (7%) of the women with lifetime eating disorders had a current eating disorder. Each diagnostic group continued to be differentiated by current eating pathology from women without lifetime eating disorders. Although approximately 75% of the women had a good outcome, less than 50% of each diagnostic group was asymptomatic. CONCLUSIONS: Eating disorders tend to improve over time often reaching subdiagnostic levels of severity, but only a minority of sufferers becomes asymptomatic. The DSM-IV diagnosis EDNOS needs to be considered in studies of the prevalence and course of eating disorders. PMID- 16476128 TI - Obsessive-compulsive spectrum of disorders: a defensible construct? AB - OBJECTIVE: To explore critically whether there is a robust basis for the concept of an obsessive-compulsive (OC) spectrum of disorders, and if so, which disorders should be included. METHOD: Selective literature review concentrating on three proposed members of the OC spectrum, namely body dysmorphic disorder, hypochondriasis and trichotillomania. RESULTS: Obsessive-compulsive disorder (OCD) itself is a heterogeneous condition or group of conditions, and this needs to be appreciated in any articulation of a 'spectrum' of OC disorders. The basis for 'membership' of the spectrum is inconsistent and varied, with varying level of support for inclusion in the putative spectrum. CONCLUSION: A more fruitful approach may be to consider behaviours and dimensions in OCD and OC spectrum disorders, and that this should be encompassed in further developments of the OC spectrum model. PMID- 16476130 TI - The prevalence of eating disorders among university students and the relationship with some individual characteristics. AB - OBJECTIVE: The purpose of this study was to determine the prevalence of eating disorders (EDs) among university students in a rural area of Turkey and to compare groups based on the sociodemographic data, history of child abuse and neglect, family roles and self-esteem with a normal control group regarding EDs. METHOD: Subjects who were chosen by simple random sampling method were consisted of 980 Cumhuriyet University students who agreed to participate out of the 1003 total students and were given a sociodemographic information form and an Eating Attitudes Test (EAT). Students who scored above a cutoff level on the EAT were interviewed using the Structured Clinical Interview for DSM-IV axis I Disorders (SCID-I), Clinical Version. The Rosenberg Self-Esteem Scale, Family Assessment Device (FAD) and Childhood Abuse and Neglect Questionnaire Form were given to subjects in the control and study groups. RESULTS: Seventy-one of the 951 students (492 female, 459 male) who correctly filled out the EAT had a score above the cutoff level of 30 or higher. Of these 71 students, 21 (2.20%) were found to have an eating disorder based on the SCID-I. No subjects were found to have anorexia nervosa. Eighteen of the 21 subjects were female. Of these 18 female students, 15 (1.57%) were found to have bulimia nervosa and three (0.31%) were found to have binge eating disorder (BED). All of the three male subjects were diagnosed with BED (0.31%). The self-esteem of those in the study group was lower than those in the control group (p < 0.001). Subjects in the study group had more frequent histories of sexual and emotional abuse in childhood (p < 0.05). Also, in the study group scores showing communication in FAD families, unity and emotional attachment were statistically significantly lower than the control group (p < 0.001). CONCLUSIONS: It has been observed from the results of this research that the frequency of bulimia nervosa and BED in this sample is so similar to Western samples. Besides that, self-esteem, child abuse and neglect, and family functions must be examined in detail because they are risk factors for EDs and affect the course of treatment. PMID- 16476131 TI - Correlates of the use of purging and non-purging methods of weight control in a community sample of women. AB - OBJECTIVE: To inform the classification of bulimic-type eating disorders, the correlates of purging and non-purging methods of weight control were examined in a large community sample of young adult women reporting recurrent episodes of binge eating. METHOD: Scores on self-report measures of eating disorder psychopathology, functional impairment and health-service utilization were compared among individuals who reported (recurrent episodes of binge eating and) the use of either purging (self-induced vomiting, laxative or diuretic misuse; n = 41) or non-purging (extreme dietary restriction, excessive exercise, or use of diet pills; n = 62) methods of weight control. Individuals who reported recurrent binge eating in the absence of extreme weight control behaviours (n = 442) were also included in the analysis. RESULTS: Non-purgers tended to be younger and heavier and have higher levels of eating disorder psychopathology and functional impairment than purgers and non-compensating binge eaters, however these differences were not statistically significant. Purgers were more likely than non purgers to have sought treatment specifically for a problem with eating, however this difference was no longer significant after age and body mass index were statistically controlled. In multivariate analysis, frequency of extreme dietary restriction was the best predictor of functional impairment. CONCLUSIONS: These findings call into question the validity of subtyping of bulimia nervosa into purging and non-purging forms as outlined in the fourth edition of the Diagnostic and Statistical Manual of Mental Disorders. PMID- 16476132 TI - Eating disorder attitudes and psychopathology in Caucasian Australian, Asian Australian and Thai university students. AB - OBJECTIVE: To examine eating disorder attitudes and psychopathology among female university students in Australia and Thailand. METHOD: Participants were 110 Caucasian Australians, 130 Asian Australians and 101 Thais in Thailand. The instruments included the Eating Attitudes Test (EAT) and the Eating Disorders Inventory (EDI). RESULTS: Eating disorder attitudes and psychopathology scores in the Thai group were found to be highest. The Asian Australian group did not have significantly higher scores on the EAT-26 than the Caucasian Australian group, but had higher scores in some subscales of the EDI-2. That the Thai group had the highest scores in susceptibility to developing an eating disorder and eating disorder psychopathology may be partially explained in sociocultural terms, with pressure to be thin more extreme in Thailand than in Australia. The evidence suggested that unhealthy eating disorder psychopathology is not limited to Western societies but is already present in Thai and other Asian societies. PMID- 16476133 TI - Quality of life: eating disorders. AB - OBJECTIVE: There is a lack of measurements with predictive validity that are specific for quality of life (QOL) in patients with eating disorders. METHOD: A total of 306 eating disorder patients treated as inpatients completed the Quality of Life for Eating Disorders (QOL ED): 109 at both admission and discharge from hospital, 65 at both admission and after 12 months. Patients also completed well validated measures of eating disorders, psychological dysfunction and general physical and mental QOL. QOL ED consists of 20 self-report questions that provide scores for the domains of behaviour, eating disorder feelings, psychological feelings, effects on daily life, effects on acute medical status and body weight, and a global score. RESULTS: QOL ED domain scores correlated appropriately with previously validated well-known measures of eating disorders, psychological dysfunction, general QOL and behaviour and body weight (p < 0.001). The QOL ED shows high reliability (Cronbach's alpha = 0.93). All scores changed significantly and appropriately during inpatient hospital treatment and between admission and 12 months after discharge from hospital (p < 0.001). The scores differed for anorexia nervosa, bulimia, eating disorder not specified (EDNOS) and no diagnosis. All no diagnosis (recovered) domain and global scores were significantly different from all diagnoses scores (p < 0.001). CONCLUSION: This quick, simple instrument fulfils all potential uses for QOL assessments in the clinical and research settings associated with eating disorders, including outcome. PMID- 16476134 TI - Illicit drug use and dependence in a New Zealand birth cohort. AB - OBJECTIVE: To describe the patterns of illicit drug use in a birth cohort studied to the age of 25 years. METHOD: The data were gathered during the Christchurch Health and Development Study. In this study a cohort of 1265 children born in the Christchurch, New Zealand urban region in mid-1977 have been studied to the age of 25 years. Information was gathered on patterns of illicit drug use and dependence during the period 15-25 years. RESULTS: By age 25 years, 76.7% of the cohort had used cannabis, while 43.5% had used other illicit drugs on at least one occasion. In addition, 12.5% of the cohort met DSM-IV criteria for dependence on cannabis, and 3.6% of the cohort met criteria for dependence on other illicit drugs at some time by age 25. There was also evidence of substantial poly-drug use among the cohort, with hallucinogens and amphetamines being the most commonly used illicit drugs (excluding cannabis). Illicit drug use and dependence was higher in males, in Maori, and in those leaving school without qualifications. Key risk factors for illicit drug use and dependence included adolescent risk taking behaviours including cigarette smoking and alcohol consumption, affiliation with substance-using peers, novelty-seeking, and conduct problems in adolescence. Other key risk factors included parental history of illicit drug use and childhood sexual abuse. CONCLUSIONS: Levels of cumulative illicit drug use in this cohort were relatively high, with the majority of respondents having tried illicit drugs by age 25. For the majority of illicit drug users, drug use did not lead to problems of dependence. Nonetheless, nearly 15% of the cohort showed symptoms of illicit drug dependence by the age of 25 years, with cannabis dependence accounting for the majority of illicit drug dependence. PMID- 16476136 TI - Brain activation during craving for alcohol measured by positron emission tomography. AB - OBJECTIVE: Craving for alcohol is probably involved in acquisition and maintenance of alcohol dependence to a substantial degree. However, the brain substrates and mechanisms that underlie alcohol craving await more detailed elucidation. METHOD: Positron emission tomography was used to map regional cerebral blood flow (CBF) in 21 detoxified patients with alcohol dependence during exposure to alcoholic and non-alcoholic beverages. RESULTS: During the alcohol condition compared with the control condition, significantly increased CBF was found in the ventral putamen. Additionally, activated areas included insula, dorsolateral prefrontal cortex and cerebellum. Cerebral blood flow increase in these regions was related to self-reports of craving assessed in the alcoholic patients. CONCLUSIONS: In this investigation, cue-induced alcohol craving was associated with activation of brain regions particularly involved in brain reward mechanisms, memory and attentional processes. These results are consistent with studies on craving for other addictive substances and may offer strategies for more elaborate studies on the neurobiology of addiction. PMID- 16476135 TI - Coexisting psychiatric disorders in a New Zealand outpatient alcohol and other drug clinical population. AB - OBJECTIVE: To describe the extent of psychiatric disorder and mental health service utilization in a representative outpatient alcohol and other drug (AOD) treatment sample in New Zealand. METHOD: A total of 105 patients were randomly recruited from two outpatient AOD treatment services in New Zealand and completed a diagnostic interview within the first 2 months of treatment. Axis I psychiatric diagnoses were made using the computerized Composite International Diagnostic Interview (CIDI-Auto), and were supplemented by the South Oaks Gambling Scale (SOGS) and the conduct disorder and antisocial personality disorder section of the Diagnostic Interview for Genetic Studies (DIGS). RESULTS: Seventy-four per cent of the sample had a current non-substance or gambling axis I disorder, with a lifetime rate of 90%. The most commonly diagnosed of these coexisting psychiatric disorders were major depressive episode (34%), social phobia (31%) and posttraumatic stress disorder (31%). Past contact with mental health services was common, while contact at the time of baseline assessment was uncommon. CONCLUSIONS: Coexisting psychiatric disorder was the rule and not the exception in this sample. AOD patients are clearly part of the larger population of mental health patients. AOD services need to be capable of comprehensive assessment and treatment planning, which includes coexisting psychiatric disorders, and should work toward better integration with other mental health services. PMID- 16476137 TI - Trauma, post-migration living difficulties, and social support as predictors of psychological adjustment in resettled Sudanese refugees. AB - OBJECTIVE: This paper explores the impact of pre-migration trauma, post-migration living difficulties and social support on the current mental health of 63 resettled Sudanese refugees. METHOD: A semistructured interview including questionnaires assessing sociodemographic information, pre-migration trauma, anxiety, depression and posttraumatic stress, post-migration living difficulties and perceived social support were administered assisted by a bilingual community worker. RESULTS: Resettled refugees from Sudan evidenced a history of trauma. Less than 5% met criteria for posttraumatic stress but 25% reported clinically high levels of psychological distress. The results indicate that social support- particularly perceived social support from the migrant's ethnic community--play a significant role in predicting mental health outcomes. Pre-migration trauma, family status and gender were also associated with mental health outcomes. CONCLUSIONS: Refugees in Australia may constitute a particularly vulnerable group in terms of mental health outcomes. Culturally specific sequelae in terms of social isolation and acculturation may be particularly problematic for these migrants. PMID- 16476138 TI - Seizure threshold determination for electroconvulsive therapy: stimulus dose titration versus age-based estimations. AB - OBJECTIVE: This study compared electroconvulsive therapy (ECT) seizure threshold determined by stimulus dose titration with age-based estimations. METHOD: Patients with major depressive disorder had their initial seizure thresholds determined by stimulus dose titration and the results compared with age-based estimations. There are no significant differences in thresholds determined by these methods. RESULTS: Two hundred and three psychiatric patients (149 females, 54 males) had their seizure thresholds determined by stimulus dose titration. There was a significant positive correlation between seizure thresholds and age for males and females with male thresholds greater than female thresholds. Age determinations of seizure threshold would have resulted in excessive initial treatment stimuli for 30% of females and 8% of males. Ineffective stimulus doses would have been given to 2% of females and 7% of males on a full age basis and 64% using a half age strategy. CONCLUSIONS: For effective high-dose right unilateral ECT, initial seizure threshold should be determined by stimulus dose titration. PMID- 16476139 TI - Stevens-Johnson syndrome (SJS) leading to resolution of mania. PMID- 16476140 TI - Persistent developmental stuttering: treatment with risperidone. PMID- 16476141 TI - Aripiprazole-induced extrapyramidal side effects in a patient with schizoaffective disorder. PMID- 16476142 TI - Weight loss during prolonged branched-chain amino acid treatment for tardive dyskinesia in a patient with schizophrenia. PMID- 16476146 TI - Pharmacological treatment in the early phase of bipolar disorders: what stage are we at? AB - OBJECTIVE: To review available guidelines, explore treatment strategies currently applied, identify critical issues and propose direction for new developments. METHOD: Literature review based on Medline search and hand search of relevant literature. RESULTS: Pharmacological treatment of the early phase of bipolar disorders lacks specific guidelines. Mood stabilizers are often prescribed after many years of illness, antipsychotic medications are frequently prescribed and often for extensive periods of time, and adherence to medication is relatively poor. In addition, mania is frequently misdiagnosed, and there is limited knowledge on which to base identification of bipolar depression and identification of the initial prodrome to bipolar disorders. CONCLUSIONS: There is an urgent need for more research in the early phases of bipolar disorders to develop means to identify patients earlier and to develop approaches that would address the specific needs of this population in a more adequate manner. PMID- 16476147 TI - Rural suicide--people or place effects? AB - OBJECTIVE: To examine factors that may contribute to elevated rates of suicide among rural communities in Australia. METHOD: A wide-ranging literature search reviewed possible factors that may contribute to the geographical variation in suicide. Literature was organized to enable examination of compositional and contextual explanations, as well as collective social functioning and social practices. RESULTS: A variety of factors may contribute to elevated rates of suicide in rural compared with urban areas. Collective and contextual (place) factors seem to be of particular importance as possible contributors to the elevated rate of suicide among rural males. These include rural socioeconomic decline; facilitators and barriers to service utilization such as service availability and accessibility, rural culture, community attitudes to mental illness and help seeking; and exposure to firearms. CONCLUSIONS: Compositional, contextual and collective factors are hypothesized to influence the elevated rate of suicide in rural compared with urban areas. These factors need to be tested in empirical studies that consider both individual and community-based risk factors, and are designed to enable exploration of likely within-rural differences. PMID- 16476148 TI - Regionally specific changes in levels of cortical S100beta in bipolar 1 disorder but not schizophrenia. AB - OBJECTIVE: To determine if levels of the glial-derived proteins S100beta and glial acidic fibrillary protein (GFAP) and the pro- and antiapoptotic proteins p53 and Bcl-2 were altered in the cortex of subjects with schizophrenia or bipolar 1 disorder. METHOD: Levels of S100beta, GFAP, p53 and Bcl-2 were measured in cortex (Brodmann's Areas (BAs) 9, 10, 46 and 40) of control subjects and subjects with schizophrenia, bipolar 1 disorder and in the cortex of rats treated with haloperidol or lithium using protein-specific antibodies and western blot analysis. RESULTS: Levels of S100beta were decreased in BA 9 and increased in BA 40 from subjects with bipolar 1 disorder. Levels of this protein were not altered in other CNS regions, in schizophrenia or in the cortex of rats treated with haloperidol or lithium. No changes in levels of the other three proteins were detected across diagnoses. CONCLUSIONS: Regionally selective changes in cortical S100beta may be associated with the pathology of bipolar 1 disorder and may reflect derangements in neuronal death or survival. PMID- 16476149 TI - Genetic, developmental and personality correlates of self-mutilation in depressed patients. AB - OBJECTIVE: To examine whether the T allele of G protein beta3 (GNbeta3) is associated with self-mutilation in depressed patients. METHOD: A history of self mutilation was systematically inquired about when recruiting depressed patients for a long-term treatment trial. Risk factors such as borderline personality disorder and childhood abuse experiences were systematically assessed, and patients were genotyped for polymorphisms of GNbeta3. RESULTS: The T allele of GNbeta3, borderline personality disorder and childhood sexual abuse were all significantly associated with self-mutilation in depressed patients. These associations were significant in both univariate and multivariate analyses, and as predicted were stronger in young depressed patients than in depressed patients of all ages. CONCLUSIONS: If the association between the T allele of GNbeta3 and self-mutilation can be replicated, this may provide clues to understanding the neurobiology of self-mutilation. PMID- 16476150 TI - Postnatal depressive symptoms among Pacific mothers in Auckland: prevalence and risk factors. AB - OBJECTIVE: To assess the prevalence of and risk factors for postnatal depressive symptoms in a cohort of mothers of Pacific Island infants in Auckland, New Zealand. METHOD: The data were gathered as part of the Pacific Island Families Study, in which 1376 mothers were interviewed when their babies were 6 weeks old. The interview included the Edinburgh Postnatal Depression Scale (EPDS). RESULTS: 16.4% of mothers were assessed as probably experiencing depression. Prevalence rates varied from 7.6% for Samoans to 30.9% for Tongans. In addition to ethnicity, risk factors identified by stepwise multiple logistic regression included low Pacific Island acculturation, first birth, stress due to insufficient food, household income less than dollar 40,000, difficulty with transport, dissatisfaction with pregnancy, birth experience, baby's sleep patterns, partner relationship and home. A large prevalence difference between Tongans and other groups remained when the effects of other risk factors were controlled statistically. CONCLUSIONS: The prevalence of depressive symptoms among Pacific mothers is at the upper end of the range typically reported. Focus on the overall rate, however, obscures substantial variation between groups. Risk factors are generally similar to those identified in previous research. IMPLICATIONS: The findings have implications for prevention and treatment and caution against assuming homogeneity within ethnic categories. Further research is required to explain differences in prevalence between Tongan and other Pacific Island groups. PMID- 16476151 TI - Actions taken to cope with depression in patients seeking specialist care. AB - OBJECTIVE: To assess the rates and perceived effectiveness of actions used to cope with depression and the factors influencing these in an outpatient sample seeking help for depression. METHOD: One hundred and seventy-six patients (74 male, 102 female) aged 16-82 years (M = 42, SD = 14.4 years) with a major depressive episode (DSM-IV criteria) were assessed using a number of measures that covered sociodemographics, history of psychiatric illness, actions taken to alleviate depression and their perceived effectiveness. Logistic regression analyses assessed age, gender and illness characteristics associated with the use and perceived effectiveness of strategies adopted to manage depression. RESULTS: Medical interventions were used and perceived to be effective in alleviating depressive symptoms by most of the sample, as were self-help and complementary therapies. Sociodemographic and illness-related characteristics had a significant influence, with younger age and having experienced an episode of anxiety disorder found to be the strongest predictors for the use of coping strategies. Being female was the strongest predictor for their effectiveness. CONCLUSION: A range of actions for depression, including medical and complementary interventions, were used and found to be helpful in a sample that had sought professional help for depression. Gender, age, depression and anxiety factors were all found to predict the use and perceived effectiveness of these strategies. PMID- 16476152 TI - Explicating links between acute coronary syndrome and depression: study design and methods. AB - OBJECTIVE: To describe a regional study seeking to replicate the suggested strong links whereby lifetime and post-coronary infarction depression are associated with a significant increase in mortality and cardiac morbidity, and consider the comparative influence of both depression and anxiety. METHOD: We detail relevant international studies and describe both the methodology as well as baseline and 1 month data from our study. RESULTS: Over a 3-year period we recruited 489 subjects admitted to a Sydney cardiac unit with an Acute Coronary Syndrome (ACS), and assessed by a range of cardiac variables and measures of current and lifetime depression. Ninety-eight per cent of the sample were assessed one month after baseline recruitment to establish depression rates. Long-term outcome reviews of mortality and morbidity and hospitalization rates are proceeding. For those subjects who were depressed in the post-ACS period and, even more so for those who had experienced lifetime depression, distinctly higher scores on anxiety variables (and lifetime caseness for anxiety disorders) were established. CONCLUSIONS: The strong interdependence between anxiety and depression in this sample of patients admitted with an ACS will allow examination of the comparative extent to which expressions of 'depression' and 'anxiety' contribute to post-ACS morbidity. PMID- 16476153 TI - Firearms legislation and reductions in firearm-related suicide deaths in New Zealand. AB - OBJECTIVE: To examine the impact of introducing more restrictive firearms legislation (Amendment to the Arms Act, 1992) in New Zealand on suicides involving firearms. METHOD: National suicide data were examined for 8 years before, and 10 years following the introduction of the legislation. RESULTS: After legislation, the mean annual rate of firearm-related suicides decreased by 46% for the total population (p < 0.0001), 66% for youth (15-24 years; p < 0.0001) and 39% for adults (> or = 25 years; p < 0.01). The fraction of all suicides accounted for by firearm-related suicides also reduced for all three populations (p < 0.0001). However, the introduction of firearms legislation was not associated with reductions in overall rates of suicide for all three populations. CONCLUSIONS: Following the introduction of legislation restricting ownership and access to firearms, firearm-related suicides significantly decreased, particularly among youth. Overall rates of youth suicide also decreased over this time but it is not possible to determine the extent to which this was accounted for by changes in firearms legislation or other causes. PMID- 16476154 TI - A comparison of Australian men with psychotic disorders remanded for criminal offences and a community group of psychotic men who have not offended. AB - BACKGROUND: People remanded into custody by the courts have a substantially higher rate of severe mental disorder than other prisoners and the general population. Knowledge of their prevalence, needs and characteristics and an analysis of pathways to care may be necessary to provide mental health care effectively and efficiently. Previous prison studies focusing on psychotic offenders have suffered from the use of instruments not validated in a forensic setting and lack of a relevant comparison group. METHOD: The Diagnostic Interview for Psychosis (DP) is a composite semi-structured standardized interview schedule. It combines social and demographic descriptors with measures of functioning adapted from the World Health Organisation Disability Assessment Schedule (DAS). The remand centre surveyed had 466 cells and is the main remand and reception centre for males for the southern region of the state of Queensland, Australia. Of the 621 men screened, 65 answered yes to at least one question in the DP and were interviewed. RESULTS: Six hundred and twenty-one remandees were screened and of these 61 were interviewed as screened positive for psychotic disorder. Thirty-five per cent had been homeless for an average of 32 weeks during the previous year. Most had had little contact with families or close friends. Eighty-one per cent were receiving no treatment at the time of offence. Seventy-eight per cent were unemployed and in receipt of a pension. Eighty per cent were dependent on alcohol, cannabis or amphetamines. Statistical issues of power are detailed in the text. CONCLUSIONS: The simplistic 'prison, hospital or community treatment' debate is misleading. Instead, the development of flexible preventative, management and accommodation services for people with severe mental disorder who have committed offences is a priority. PMID- 16476155 TI - Mental disorders in female prisoners. AB - OBJECTIVE: The objective of the study was to investigate the rates of mental disorder among women in prison in Victoria, and to compare with community rates. DESIGN: A midnight census of all women in prison in Victoria was undertaken. Respondents were interviewed with a version of the Composite International Diagnostic Interview (CIDI), an adapted version of the Personality Diagnostic Questionnaire (PDQ-4+) and a demographics questionnaire. MAIN OUTCOME MEASURES: Twelve-month prevalence rates of ICD-10 mental disorders including depressive disorders, anxiety disorders and drug-related disorders were examined. Prevalence of personality disorders was also investigated. RESULTS: Eighty-four per cent of the female prisoners interviewed met the criteria for a mental disorder (including substance harmful use/dependence) in the year prior to interview. This rate was reduced to 66% when drug-related disorders were excluded. Forty-three per cent of subjects were identified as cases on a personality disorder screener. For all disorders, (except obsessive-compulsive disorder and alcohol harmful use) women in prison had a significantly greater likelihood of having met the 12-month diagnostic criteria when compared to women in the community. The most prevalent disorders among the female prisoners were: drug use disorder (57%), major depression (44%), Posttraumatic stress disorder (36%), and personality disorders. Almost a quarter (24%) of respondents were identified as a 'case' on the psychosis screen. CONCLUSIONS: In the present study female prisoners had significantly higher rates of the mental disorders investigated (with the exceptions of OCD and alcohol harmful use) when compared with women in the community. The pattern of disorder found among female prisoners is consistent with the abuse literature, suggesting that histories of abuse among the prison population may account for part of the discrepancy. These results highlight the need for improved assessment and treatment resources to meet the demands of this population. PMID- 16476156 TI - Mental disorders in Australian prisoners: a comparison with a community sample. AB - BACKGROUND: The plight of those with mental health problems and the possible role of prisons in "warehousing" these individuals has received considerable media and political attention. Prisoners are generally excluded from community-based surveys and to date no studies have compared prisoners to the community. OBJECTIVE: The objective was to examine whether excess psychiatric morbidity exists in prisoners compared to the general community after adjusting for demographics. METHOD: Prison data were obtained from a consecutive sample of reception prisoners admitted into the state's correctional system in 2001 (n = 916). Community data were obtained from the 1997 Australian National Survey of Mental Health and Wellbeing (n = 8168). Mental health diagnoses were obtained using the Composite International Diagnostic Interview and a number of other screening measures. Weighting was used in calculating the 12-month prevalence estimates to control for demographic differences between the two samples. Logistic regression adjusting for age, sex and education was used to compare the prison and community samples. RESULTS: The 12-month prevalence of any psychiatric illness in the last year was 80% in prisoners and 31% in the community. Substantially more psychiatric morbidity was detected among prisoners than in the community group after accounting for demographic differences, particularly symptoms of psychosis (OR = 11.8, 95% CI 7.5-18.7), substance use disorders (OR = 11.4, 95% CI 9.7-13.6) and personality disorders (OR = 8.6, 95% CI 7.2-10.3). Mental functioning and disability score were worse for prisoners than the community except for physical health. CONCLUSIONS: This study found an overrepresentation of psychiatric morbidity in the prisoner population. Identifying the causes of this excess requires further investigation. PMID- 16476161 TI - Idiopathic sclerosing encapsulating peritonitis (or abdominal cocoon). AB - BACKGROUND: Idiopathic sclerosing encapsulating peritonitis (or abdominal cocoon) is a rare cause of small bowel obstruction, especially in adult population. Diagnosis is usually incidental at laparotomy. We discuss one such rare case, outlining the fact that an intra-operative surprise diagnosis could have been facilitated by previous investigations. CASE PRESENTATION: A 56 year-old man presented in A&E department with small bowel ileus. He had a history of 6 similar episodes of small bowel obstruction in the past 4 years, which resolved with conservative treatment. Pre-operative work-up did not reveal any specific etiology. At laparotomy, a fibrous capsule was revealed, in which small bowel loops were encased, with the presence of interloop adhesions. A diagnosis of abdominal cocoon was established and extensive adhesiolysis was performed. The patient had an uneventful recovery and follow-up. CONCLUSION: Idiopathic sclerosing encapsulating peritonitis, although rare, may be the cause of a common surgical emergency such as small bowel ileus, especially in cases with attacks of non-strangulating obstruction in the same individual. A high index of clinical suspicion may be generated by the recurrent character of small bowel ileus combined with relevant imaging findings and lack of other plausible etiologies. Clinicians must rigorously pursue a preoperative diagnosis, as it may prevent a "surprise" upon laparotomy and result in proper management. PMID- 16476160 TI - Impact of briefly-assessed depression on secondary prevention outcomes after acute coronary syndrome: a one-year longitudinal survey. AB - BACKGROUND: Patients with acute coronary syndromes (ACS) are at increased risk of further acute cardiac events. Secondary prevention aims to decrease morbidity and mortality post-ACS. Depression is related to increased risk in this population, and to poorer secondary prevention activities. However, lengthy depression assessment techniques preclude depression assessment in routine care. The present study investigated the relationship of briefly-assessed depression with secondary prevention outcomes one year post-ACS. METHODS: Following ethics committee approval, hospitals recruited patients for a national survey of ACS. Consenting patients with ACS completed a brief depression scale during hospitalisation. The predictive validity of two brief scales was independently assessed, with groups combined for the overall sample. Participants then completed a one-year longitudinal follow-up postal survey of secondary prevention activities. RESULTS: The response rate for follow-up was 86% (n = 681). Proportions taking anti platelet (88% v 87%; p = 0.334) and lipid-lowering (83% v 84%; p = 0.437) therapies remained unchanged. Prevalence of smoking (40% v 22%; p < 0.001), and median number of cigarettes smoked (20 v 10; p < 0.001) were significantly reduced at one year. Fifty-six per cent of patients reported attending cardiac rehabilitation programmes. Of those aged < 65 years at baseline, 54% had returned to work at one year. A majority (56%) reported feeling physically better. Prevalence of depression was unchanged in those who completed a depression scale at both time points (15% v 17%; p = 0.434). Baseline depression did not predict taking anti-platelet, blood pressure or cholesterol medications (all p > 0.05), but did predict continuation of smoking (OR = 2.3, 95% CI 1.3-4.0, p = 0.003), a higher (above median) number of general practitioner visits (OR = 2.1, 95% CI 1.3 3.4, p = 0.005), failure to return to work (OR = 0.4, 95% CI 0.2-0.8, p = 0.015), and not feeling better (OR = 0.6, 95% CI 0.3-1.0, p = 0.05) at one year. CONCLUSION: Rapid depression assessment can be used to help identify patients with ACS at risk of a range of poorer secondary prevention outcomes. The results provide support for the routine screening of depression in acute settings. Strategies to increase rates of smoking cessation, return to work, general well being and decrease health service use by depressed patients may need to incorporate some element of treatment for depression. PMID- 16476163 TI - Canine parvovirus-like particles, a novel nanomaterial for tumor targeting. AB - Specific targeting of tumor cells is an important goal for the design of nanotherapeutics for the treatment of cancer. Recently, viruses have been explored as nano-containers for specific targeting applications, however these systems typically require modification of the virus surface using chemical or genetic means to achieve tumor-specific delivery. Interestingly, there exists a subset of viruses with natural affinity for receptors on tumor cells that could be exploited for nanotechnology applications. For example, the canine parvovirus (CPV) utilizes transferrin receptors (TfRs) for binding and cell entry into canine as well as human cells. TfRs are over-expressed by a variety of tumor cells and are widely being investigated for tumor-targeted drug delivery. We explored whether the natural tropism of CPV to TfRs could be harnessed for targeting tumor cells. Towards this goal, CPV virus-like particles (VLPs) produced by expression of the CPV-VP2 capsid protein in a baculovirus expression system were examined for attachment of small molecules and delivery to tumor cells. Structural modeling suggested that six lysines per VP2 subunit are presumably addressable for bioconjugation on the CPV capsid exterior. Between 45 and 100 of the possible 360 lysines/particle could be routinely derivatized with dye molecules depending on the conjugation conditions. Dye conjugation also demonstrated that the CPV-VLPs could withstand conditions for chemical modification on lysines. Attachment of fluorescent dyes neither impaired binding to the TfRs nor affected internalization of the 26 nm-sized VLPs into several human tumor cell lines. CPV-VLPs therefore exhibit highly favorable characteristics for development as a novel nanomaterial for tumor targeting. PMID- 16476164 TI - HIV/AIDS prevalence and behaviour in drug users and pregnant women in Kashgar Prefecture: case report. AB - Second Generation sub-population HIV Surveillance was undertaken in Kashgar City and Shache County, Xinjiang, Peoples Republic of China between December 2003 and January 2004, targeting injecting and mixed method drug users and pregnant and postnatal women. The study aimed to determine the extent to which the epidemic is shifting from a concentrated stage to a more generalised epidemic. One hundred and forty two (142) exclusively injecting drug users (66) and mixed method drug users (injecting and non-injecting-76) participated in this survey. Eight hundred and two (802) pregnant and postnatal women participated in the survey. In Kashgar City and Shache County the serum prevalence of HIV amongst injecting drug users was 56.06%, for mixed method drug users 48.68% and 0.38% in pregnant women. In Shache County HIV infection rates were significantly lower in drug user groups and amongst pregnant and post-natal women, at 2.22% and 0% respectively. The behavioral survey indicated that 15% of injecting drug users have shared needles (however sero prevalence and knowledge in relation to access to clean needles and syringes suggests that this may not reflect the actual situation). Knowledge of prevention of transmission strategies (not sharing needles and condom utilisation) is similar between both groups at 60-70%. However it appears that this knowledge has not significantly impacted on behavior such as needle sharing and condom utilisation. In Kashgar City and Shache County there have been very few interventions to support HIV/AIDS prevention, care and control. The results from this survey will inform future directions and the development and implementation of targeted interventions including targeted information dissemination and harm reduction strategies. This survey was funded by the Xinjiang HIV/AIDS Prevention and Care Project, a bilateral project jointly implemented by the Government of the People's Republic of China and the Government of Australia. PMID- 16476165 TI - RibAlign: a software tool and database for eubacterial phylogeny based on concatenated ribosomal protein subunits. AB - BACKGROUND: Until today, analysis of 16S ribosomal RNA (rRNA) sequences has been the de-facto gold standard for the assessment of phylogenetic relationships among prokaryotes. However, the branching order of the individual phlya is not well resolved in 16S rRNA-based trees. In search of an improvement, new phylogenetic methods have been developed alongside with the growing availability of complete genome sequences. Unfortunately, only a few genes in prokaryotic genomes qualify as universal phylogenetic markers and almost all of them have a lower information content than the 16S rRNA gene. Therefore, emphasis has been placed on methods that are based on multiple genes or even entire genomes. The concatenation of ribosomal protein sequences is one method which has been ascribed an improved resolution. Since there is neither a comprehensive database for ribosomal protein sequences nor a tool that assists in sequence retrieval and generation of respective input files for phylogenetic reconstruction programs, RibAlign has been developed to fill this gap. RESULTS: RibAlign serves two purposes: First, it provides a fast and scalable database that has been specifically adapted to eubacterial ribosomal protein sequences and second, it provides sophisticated import and export capabilities. This includes semi-automatic extraction of ribosomal protein sequences from whole-genome GenBank and FASTA files as well as exporting aligned, concatenated and filtered sequence files that can directly be used in conjunction with the PHYLIP and MrBayes phylogenetic reconstruction programs. CONCLUSION: Up to now, phylogeny based on concatenated ribosomal protein sequences is hampered by the limited set of sequenced genomes and high computational requirements. However, hundreds of full and draft genome sequencing projects are on the way, and advances in cluster-computing and algorithms make phylogenetic reconstructions feasible even with large alignments of concatenated marker genes. RibAlign is a first step in this direction and may be particularly interesting to scientists involved in whole genome sequencing of representatives of new or sparsely studied eubacterial phyla. RibAlign is available at http://www.megx.net/ribalign. PMID- 16476166 TI - Regular use of a hand cream can attenuate skin dryness and roughness caused by frequent hand washing. AB - BACKGROUND: Aim of the study was to determine the effect of the regular use of a hand cream after washing hands on skin hydration and skin roughness. METHODS: Twenty-five subjects washed hands and forearms with a neutral soap four times per day, for 2 minutes each time, for a total of two weeks. One part of them used a hand cream after each hand wash, the others did not (cross over design after a wash out period of two weeks). Skin roughness and skin hydration were determined on the forearms on days 2, 7, 9 and 14. For skin roughness, twelve silicon imprint per subject and time point were taken from the stratum corneum and assessed with a 3D skin analyzer for depth of the skin relief. For skin hydration, five measurements per subject and time point were taken with a corneometer. RESULTS: Washing hands lead to a gradual increase of skin roughness from 100 (baseline) to a maximum of 108.5 after 9 days. Use of a hand cream after each hand wash entailed a decrease of skin roughness which the lowest means after 2 (94.5) and 14 days (94.8). Skin hydration was gradually decreased after washing hands from 79 (baseline) to 65.5 after 14 days. The hand wash, followed by use of a hand cream, still decreased skin hydration after 2 days (76.1). Over the next 12 days, however, skin hydration did not change significantly (75.6 after 14 days). CONCLUSION: Repetitive and frequent hand washing increases skin dryness and roughness. Use of a hand cream immediately after each hand wash can confine both skin dryness and skin roughness. Regular use of skin care preparations should therefore help to prevent both dry and rough skin among healthcare workers in clinical practice. PMID- 16476167 TI - Occupation, smoking, and chronic obstructive respiratory disorders: a cross sectional study in an industrial area of Catalonia, Spain. AB - BACKGROUND: Few studies have investigated the independent effects of occupational exposures and smoking on chronic bronchitis and airflow obstruction. We assessed the association between lifetime occupational exposures and airflow obstruction in a cross-sectional survey in an urban-industrial area of Catalonia, Spain. METHODS: We interviewed 576 subjects of both sexes aged 20-70 years (response rate 80%) randomly selected from census rolls, using the ATS questionnaire. Forced spirometry was performed by 497 subjects according to ATS normative. RESULTS: Lifetime occupational exposure to dust, gases or fumes was reported by 52% of the subjects (63% in men, 41% in women). Textile industry was the most frequently reported job in relation to these exposures (39%). Chronic cough, expectoration and wheeze were more prevalent in exposed subjects with odds ratios ranging from 1.7 to 2.0 being highest among never-smokers (2.1 to 4.3). Lung function differences between exposed and unexposed subjects were dependent on duration of exposure, but not on smoking habits. Subjects exposed more than 15 years to dusts, gases or fumes had lower lung function values (FEV1 -80 ml, 95% confidence interval (CI) -186 to 26; MMEF -163 ml, CI -397 to 71; FEV1/FVC ratio 1.7%, CI -3.3 to -0.2) than non-exposed. CONCLUSION: Chronic bronchitis symptoms and airflow obstruction are associated with occupational exposures in a population with a high employment in the textile industry. Lung function impairment was related to the duration of occupational exposure, being independent of the effect of smoking. PMID- 16476168 TI - Antibiogram pattern and seasonality of Salmonella serotypes in a North Indian tertiary care hospital. AB - The antibiogram pattern and seasonal distribution of Salmonella serotypes were analysed retrospectively over a 6-year period from January 1999 to December 2004. Blood cultures received in the Bacteriology Laboratory were processed by standard procedures and the Salmonella spp. isolates were identified with specific antisera and standard biochemical tests. Antimicrobial susceptibility testing was carried out by a standard disc diffusion method and the minimum inhibitory concentration (MIC) of ciprofloxacin for 332 representative Salmonella isolates was determined by E test. Salmonella Typhi (75.7%) was the predominant serotype among 830 Salmonella spp. isolated during the study period followed by S. Paratyphi A (23.8%). The maximum number of enteric fever cases occurred during April-June (dry season) followed by July-September (monsoon season). There was a decrease in multidrug-resistant (MDR) S. Typhi, but MDR S. Paratyphi A isolates increased. There was also a dramatic increase in nalidixic acid-resistant isolates. All isolates were susceptible to third-generation cephalosporins and ciprofloxacin except one S. Typhi strain which demonstrated high-level ciprofloxacin resistance with a MIC of 16 mug/ml. A knowledge of the seasonal distribution and antibiotic resistance pattern of Salmonella in a particular geographical region is helpful in the delineation of appropriate control measures required for prevention of enteric fever. PMID- 16476169 TI - Evaluation of measures to reduce international spread of SARS. AB - Mathematical models are used to quantify the effect of border control measures in reducing the international spread of SARS. Border screening is shown to play a relatively minor role in reducing disease spread. Assuming detection rates similar to those reported for arrival screening in Australia, screening can detect up to 10% (95% CI 3-23) of infected travellers, and reduce the probability of a large outbreak by up to 7% (95% CI 2-17). Rapid reductions in the time to diagnosis and effective facilities for the isolation of cases are essential to ensure that there will not be a large outbreak, and each week of delay in responding to imported infection approximately doubles the total number of cases. While the control response is being developed in a currently uninfected region, border screening can provide up to one week's additional time in which to improve methods for early isolation of cases. PMID- 16476170 TI - Modelling the transmission of airborne infections in enclosed spaces. AB - The Wells-Riley equation for modelling airborne infection in indoor environments is incorporated into an SEIR epidemic model with a short incubation period to simulate the transmission dynamics of airborne infectious diseases in ventilated rooms. The model enables the effect of environmental factors such as the ventilation rate and the room occupancy to be examined, and allows the long-term impact of infection control measures to be assessed. A theoretical parametric study is carried out to demonstrate how changes to both the physical environment and infection control procedures may potentially limit the spread of short incubation-period airborne infections in indoor environments such as hospitals. PMID- 16476171 TI - Evaluation of the meningitis epidemics risk model in Africa. AB - Meningitis epidemics have a strong environmental component in Africa with the most severe epidemics occurring in the Sahelian region known as the Meningitis Belt. The objective of this study is to evaluate an ecological model based on absolute humidity and land cover type to predict the location of these epidemics. The risk model is evaluated prospectively based on epidemics occurring in Africa from January 2000 to April 2004. Seventy-one epidemics occurred during this time period affecting 22% of continental African districts. The model predicted their location with a sensitivity of 88%. The evaluation also suggests that epidemics may be extending south of the Sahel, which is consistent with environmental changes in the region. This model could be used to select priority areas for the introduction of the newly developed conjugate meningococcal vaccines. Further studies are needed to enhance our understanding of the complex relationship between meningitis epidemics and the environment. PMID- 16476173 TI - The effect of pre-milking teat-brushing on milk processing time in an automated milking system. AB - Cow throughput in an automatic milking system (AMS) is limited by system parameters such as the time required for pre-milking udder preparation and cup attachment, physiological responses of the cow (such as milk let-down and milking out rate), milking machine features and cow behaviour. A single-factor cross-over design was used to investigate the effect of pre-milking teat brushing on milk processing time in an AMS operating in an extensive grazing farming system. Teat brushing consisted of two roller brushes tracking up each teat three times (total brushing time of up to 45 s/cow). Cows were allocated to one of two treatment groups with either no brushing (NB) or brushing (B) for a 4-week period before being changed to the other treatment. Teat brushing resulted in shorter average cups-on-time (B = 506.1 s, NB = 541.0 s, P = 0.0001), longer average milk processing time (B = 10.30 min, NB = 9.76 min, P = 0.001) and no difference in daily milk yield (B = 14.67, NB = 14.71 kg/cow, P = 0.826). There was no difference between the two treatments in the success of cup attachment (B = 3.76%, NB = 5.10% unsuccessful milking attempts, P = 0.285). The estimated time cost of pre-milking teat brushing was 53 min for every 100 milkings, equivalent to an additional 5-6 milkings for every 100 milkings by an AMS. The importance of these potential time savings is discussed in relation to automatic milking in farming systems that aim for a lower per cow milking frequency and high ratio of cows to AMS. PMID- 16476172 TI - Peptides released from acid goat whey by a yeast-lactobacillus association isolated from cheese microflora. AB - Seven lactobacilli and a variety of microflora extracted from twenty five commercial cheeses were grown on unsupplemented acid goat whey and screened for their capacity to hydrolyse whey proteins [alpha-lactalbumin (alpha-la) and beta lactoglobulin (beta-lg)] and to generate peptides. Fermentations were performed aerobically or anaerobically at 37 degrees C using crude or pre-heated whey (10 min at 65, 75 or 85 degrees C). Under aerobic conditions, growth of lactobacilli was poor and protein hydrolysis did not occur. Anaerobic conditions slightly increased lactobacilli growth but neither beta-lg hydrolysis nor peptide generation were observed. More than 50% of alpha-la was digested into a truncated form of alpha-la (+/- 12 kDa) in crude whey and whey pre-heated at 65 degrees C. Twenty-five microflora extracted from raw milk cheeses were screened for their proteolytic activities on acid goat whey under the conditions previously described. Eight of them were able to hydrolyse up to 50% of alpha-la mainly during aerobic growth on crude or pre-heated whey. The corresponding hydrolysates were enriched in peptides. The hydrolysate involving microflora extracted from Comte cheese after or at 18 months ripening was the only one to exhibit hydrolysis of both alpha-la and beta-lg. Microbiological analysis showed that microorganisms originating from Comte cheese and capable of growth on unsupplemented whey consisted of Candida parapsilosis and Lactobacillus paracasei. Fermentation kinetic profiles suggested that peptides were released from alpha-la hydrolysis. The co-culture of both microorganisms was required for alpha-la hydrolysis that occurred concomitantly with the pH decrease. During whey fermentation, Cand. parapsilosis excrete at least one protease responsible for alpha-la hydrolysis, and Lb. paracasei is responsible for medium acidification that is required for protease activation. PMID- 16476162 TI - EST analysis of the scaly green flagellate Mesostigma viride (Streptophyta): implications for the evolution of green plants (Viridiplantae). AB - BACKGROUND: The Viridiplantae (land plants and green algae) consist of two monophyletic lineages, the Chlorophyta and the Streptophyta. The Streptophyta include all embryophytes and a small but diverse group of freshwater algae traditionally known as the Charophyceae (e.g. Charales, Coleochaete and the Zygnematales). The only flagellate currently included in the Streptophyta is Mesostigma viride Lauterborn. To gain insight into the genome evolution in streptophytes, we have sequenced 10,395 ESTs from Mesostigma representing 3,300 independent contigs and compared the ESTs of Mesostigma with available plant genomes (Arabidopsis, Oryza, Chlamydomonas), with ESTs from the bryophyte Physcomitrella, the genome of the rhodophyte Cyanidioschyzon, the ESTs from the rhodophyte Porphyra, and the genome of the diatom Thalassiosira. RESULTS: The number of expressed genes shared by Mesostigma with the embryophytes (90.3 % of the expressed genes showing similarity to known proteins) is higher than with Chlamydomonas (76.1 %). In general, cytosolic metabolic pathways, and proteins involved in vesicular transport, transcription, regulation, DNA-structure and replication, cell cycle control, and RNA-metabolism are more conserved between Mesostigma and the embryophytes than between Mesostigma and Chlamydomonas. However, plastidic and mitochondrial metabolic pathways, cytoskeletal proteins and proteins involved in protein folding are more conserved between Mesostigma and Chlamydomonas than between Mesostigma and the embryophytes. CONCLUSION: Our EST-analysis of Mesostigma supports the notion that this organism should be a suitable unicellular model for the last flagellate common ancestor of the streptophytes. Mesostigma shares more genes with the embryophytes than with the chlorophyte Chlamydomonas reinhardtii, although both organisms are flagellate unicells. Thus, it seems likely that several major physiological changes (e.g. in the regulation of photosynthesis and photorespiration) took place early during the evolution of streptophytes, i.e. before the transition to land. PMID- 16476174 TI - Welfare effects of the use of recombinant bovine somatotropin in the USA. AB - The welfare effects of increased milk production associated with the use of recombinant bovine somatotropin (rBST) on dairy operations in the USA were examined for 1996. Results that derived from three different estimates of the milk-production response to rBST were evaluated and compared. One estimate, derived from a survey of dairy producers in Connecticut, led to economic-impact estimates that were not statistically significant. A second, derived from a national survey that concentrated on the health and management of dairy cattle, led to estimates that were unbelievably high. A third, derived from a national survey that concentrated on the economics of dairy producers, provided the most reasonable estimates of economic impacts. Results of economic analysis, using the latter results, indicated that if rBST had not caused milk production to increase, then the market price of milk would have been 2.2 +/- 1.5 cents/kg higher, and the total value of the milk produced would have risen from Dollars 23.0 +/- 0.6 billion to Dollars 24.1 +/- 1.0 billion. A welfare analysis demonstrated that the increased milk production (and the reduced market price) associated with the use of rBST in the USA caused the economic surplus of consumers to rise by Dollars 1.5 +/- 1.0 billion, while the economic surplus of dairy producers fell by Dollars 1.1 +/- Dollars 0.8 billion. Increased milk production associated with rBST yielded a total gain to the US economy of Dollars 440 +/- 280 million. An analysis of annual percent changes in the number of dairy cows per operation, milk production per cow, total milk production, total number of dairy cows, and total number of dairy operations in the USA suggested that the dairy industry's long-term economic growth path was stable from 1989-2001 inclusive, and did not receive a shock resulting from the introduction of rBST. PMID- 16476175 TI - Effects of dietary protein level on ewe milk yield and nitrogen utilization, and on air quality under different ventilation rates. AB - The experiment, which lasted 53 d, was conducted during the winter (February and March) of 2004 and used 48 Comisana ewes in mid lactation. A 2 x 2 factorial design was used, with ewes receiving two levels of dietary crude protein (CP) (moderate, 16% CP v. low, 13% CP) in the dry matter (DM) and being exposed to two ventilation rates (moderate, 47 m3/h v. low, 23.5 m3/h per ewe) for each dietary treatment. Air concentrations of NH3 and of microorganisms were measured twice weekly. Milk yield was recorded daily. Individual milk samples were analysed weekly for composition and fortnightly for bacteriological characteristics. After the last milk sampling (day 49 of the study period), four animals from each group were placed in a metabolism box and their individual faeces and urine were collected for three consecutive days. Amounts of urine and faeces excreted, and urinary and faecal N outputs were measured. The 16% CP diet resulted in a lower milk casein content and a higher milk urea concentration than the 13% CP diet, as well as in a reduced gross efficiency of utilization of dietary N, a greater amount of N excreted and a higher total coliform concentration in milk. The moderate ventilation rate resulted in higher yields of milk, irrespective of CP content. Significant interactions of CP level x ventilation rate were found for the amounts of urine, of total water and of faecal N, and for mesophilic concentration in milk, the highest values being displayed by the ewes fed the 16% CP diet and exposed to the low ventilation rate. The moderate dietary CP level and low ventilation rate had a deleterious effect on air concentrations of microorganisms and ammonia. Results suggested that a reduction of dietary CP level from 16 to 13% of DM had no detrimental effect on ewe milk yield in mid lactation and could even improve some of its nutritional and hygienic characteristics. Our findings also indicate that the choice of a proper ventilation rate is critical for high efficiency of production in the lactating ewe, especially in intensively managed flocks receiving diets high in CP. PMID- 16476176 TI - Effects of length of dry period on yields of milk fat and protein, fertility and milk somatic cell score in the subsequent lactation of dairy cows. AB - The objective was to utilize data from modern US dairy cattle to determine the effect of days dry on fat and protein yield, fat and protein percentages, days open, and somatic cell score in the subsequent lactation. Field data collected through the dairy herd improvement association from January 1997 to December 2003 and extracted from the Animal Improvement Programs Laboratory national database were used for analysis. Actual lactation records calculated from test-day yields using the test-interval method were used in this study. The model for analyses included herd-year of calving, year-state-month of calving, previous lactation record, age at calving, and days dry as a categorical variable. Fat and protein yield was maximized in the subsequent lactation with a 60-d dry period. Dry periods of 20 d or less resulted in substantial losses in fat and protein yield in the subsequent lactation. In contrast to yields, a short dry period was beneficial for fat and protein percentages. Short dry periods also resulted in fewer days open in the subsequent lactation; however, this was entirely due to the lower milk yield associated with shortened dry period. When adjusted for milk yield, short dry periods actually resulted in poorer fertility in the subsequent lactation. Long days dry improved somatic cell score in the subsequent lactation. Herds with mastitis problems should be cautious in shortening days dry because short dry periods led to higher cell scores in the subsequent lactation compared with 60-d dry. PMID- 16476177 TI - Performance of two commonly used angiotensin-converting enzyme inhibition assays using FA-PGG and HHL as substrates. AB - In development of fermented dairy products and protein hydrolysates with high inhibitory activity towards angiotensin-converting enzyme (ACE), it is crucial to have a reliable assay for measuring the ACE activity. In the present study, the performance of two commonly used assays based on synthetic N-derivates of tripeptides as substrates were tested with respect to reliability in determination of ACE activity per se and to the inhibitory effect of a tryptic whey protein digest and captopril. In one test, the ACE activity was calculated from the amount of hippuric acid liberated from hippuryl-His-Leu (HHL) during 30 min of incubation with ACE, as quantified after HPLC separation of reaction products. In the other assay, the ACE activity was measured directly as the rate of decrease in the absorbance at 340 nm during the first 30 min of ACE catalysed hydrolysis of furanacroloyl-Phe-Glu-Glu (FA-PGG). Both assays, in the absence of inhibitor, showed a good performance with relative standard deviation between replicate samples around 7%. In the presence of inhibitor solutions, relative standard deviations for both assays varied between 1 and 18% for the variously diluted inhibitors. Both assays gave values for the concentration of inhibitor needed to inhibit ACE by 50% similar to those previously reported for whey protein digests and captopril. Different results from the two assays, however, emphasize the importance of controlling the actual ACE-activity for comparison between assays. The limitations of each assay are discussed. Considering the fewer steps in the assay using FA-PGG as substrate, and thus less time and chemicals consumed per sample, and the simpler equipment needed, this assay is recommended for the screening of clear peptide samples. PMID- 16476178 TI - A single nucleotide polymorphism in the bovine beta-casein promoter region across different bovine breeds. PMID- 16476179 TI - Composition, coagulation properties and Parmigiano-Reggiano cheese yield of Italian Brown and Italian Friesian herd milks. AB - The authors report the results of a study aimed at the comparison of the basic chemical composition, the main protein fractions distribution, rennet coagulation properties and Parmigiano-Reggiano cheese yield of vat milk from Italian Brown and Italian Friesian herds. Parmigiano-Reggiano cheese factories which manufacture milk separately from Italian Brown herds and Italian Friesian herds were used in the study. Thirteen cheesemaking trials were performed at 10 different commercial cheese factories. The study was carried out from March to October 2003. For each cheesemaking trial in each factory, approximately 1100 kg milk from Italian Brown cows and from Italian Friesian cows were processed in parallel. The animals involved in the study came from farms with comparable management practices, size, location, number of lactation and days in milking. Each vat contained milk obtained by combining milk collected during the evening milking (partially skimmed milk by natural creaming) and the following morning milking (full-cream milk), from at least 2 dairy herds. Milk from Italian Brown cows is characterised by a higher casein content (27.1 v. 23.7 g/kg; P < or = 0.0001) than Italian Friesian cows' milk. Curd firming time (k20) of Italian Brown cows' milk was markedly lower than that of Italian Friesian cows' milk (6.6 v. 10.0 min; P < or = 0.001). This implies a higher rate of aggregation of para casein micelles for Italian Brown cows' milk. The coagulum of Italian Brown cows' milk had better rheological properties and lower losses of fat in the cheese whey. Parmigiano-Reggiano cheese yield at 24 h was also higher for Italian Brown cows' milk, + 0.99 kg cheese for every 100 kg vat milk. PMID- 16476180 TI - Effect of omitting one or two milkings weekly on lactational performance in dairy ewes. AB - We investigated the effects of omitting one or two milkings weekly on milk yield and milk composition in dairy sheep. Ninety Spanish Assaf ewes were allocated to three experimental treatments: T0 (no milking omission; 14 milkings/week), T1 (omission of one evening milking per week; 13 milkings/week) and T2 (omission of two evening milkings per week; 12 milkings/week). The experiment was extended for 11 weeks, from week 7 after lambing to week 17 of lactation. Once a week, daily milk production was recorded just before and after the omission. Milk quality and composition was analysed in lactation weeks 9, 13 and 16. Omission of one or two evening milkings each week resulted in an increase in milk production recorded the day immediately after the omission (27% for T1 and 16% for T2), which can be attributed to accumulation of milk in the udder cisterns. This increase did not compensate completely for the loss of yield in the omitted milking. The lack of significant differences among the three milking strategies in milk production (on average 1903, 2062 and 1833 ml/d for T0, T1 and T2, respectively) recorded before omission throughout the trial, would indicate the absence of residual effects of the omission on production in subsequent weeks. Treatment T2 resulted in a loss of milk production of approximately 39% during those days when milkings were omitted, representing a decrease of approximately 10% of the estimated weekly milk production. In relation to milk composition, milking omission led to significant increases in the fat and protein concentrations of the milk collected after the omission, whereas differences were not significant for the milk obtained before the omission. Despite the slight increase observed in treatment T2 after the milking omission, the somatic cell count was always far below that considered as indicative of possible pathologies. In conclusion, at least one evening milking could be omitted each week in high-producing dairy sheep without adversely affecting milk yield and milk composition. PMID- 16476181 TI - Relationship between cellular and whey components in buffalo milk. AB - High somatic cell count (SCC) affects milk quality and cheesemaking, resulting in a reduction in cheese yield and quality. In dairy cows, quarter milk samples with > 200,000 cells/ml are considered to have subclinical mastitis, while there is much uncertainty on the corresponding levels of SCC in buffalo milk. In this study 30 lactating water buffaloes were selected and SCC, differential somatic cell counts and several whey components were tested in quarter milk samples to assess the relationship between inflammation markers and milk quality. Overall 236 quarter milk samples were considered. To evaluate the relationship between cellular markers (SCC, polymorphonuclear leucocytes, PMN, and N-Acetyl-beta glucosaminidase, NAGase) and other milk components, three classes were defined (low, medium and high). Analysis of milk yield showed a significant reduction in the high class of each of the three markers chosen. Overall, the highest class was characterized by significant changes in milk composition and a lower milk quality. The presence of an inflammatory status of the udder was frequent after the first trimester of lactation and in buffaloes with two or more parturitions. This study showed that significant changes in milk components can be observed when SCC are > 400,000 cells/ml, PMN are > 50% and NAGase is > 100 units. These thresholds could be suggested as levels to define udder health status in buffalo cows. PMID- 16476182 TI - Effect of high pressure homogenisation of milk on cheese yield and microbiology, lipolysis and proteolysis during ripening of Caciotta cheese. AB - The principal aim of this work was to compare Caciotta cheeses obtained from cow milk previously subjected to high pressure homogenisation (HPH) at 100 MPa with those produced from raw (R) or heat-treated (P) cow milk. HPH had both direct and indirect effects on cheese characteristics and their evolution during ripening. In particular, HPH treatment of milk induced a significant increase of the cheese yield; moreover, it affected the microbial ecology of both curd and cheese. Compared with the thermal treatment, the HPH treatment resulted in a decrease of about one log cfu/g of yeast and lactobacilli cell loads of the curd. The initial milk treatment also affected the evolution over time and the levels attained at the end of ripening of all the microbial groups studied. In fact, lactobacilli, microstaphylococci and yeast cell loads remained at lower levels in the cheeses obtained from HPH milk with respect to the other cheese types over the whole ripening period. Moreover, HPH of milk induced marked and extensive lipolysis. Cheeses from HPH milk showed the presence of high amounts of free fatty acids immediately after brining. The electrophoretic patterns of the different cheese types showed that Caciotta made from HPH-treated milk was characterized by a more extensive and faster proteolysis as well as a significant modification of its volatile molecule profile. The results obtained and the sensory analysis indicated that HPH treatment of milk was able to differentiate Caciotta cheese or to modify its ripening patterns. PMID- 16476183 TI - Real-time PCR investigation of parasite ecology: in situ determination of oyster parasite Perkinsus marinus transmission dynamics in lower Chesapeake Bay. AB - Perkinsus marinus is a severe pathogen of the oyster Crassostrea virginica on the East Coast of the United States. Transmission dynamics of this parasite were investigated in situ for 2 consecutive years (May through October) at 2 lower Chesapeake Bay sites. Compared to previous studies where seasonal infection patterns in oysters were measured, this study also provided parasite water column abundance data measured using real-time PCR. As previously observed, salinity and temperature modulated parasite transmission dynamics. Using regression analysis, parasite prevalence, oyster mortalities and parasite water column abundance were significantly positively related to salinity. Perkinsus marinus weighted prevalence in wild oysters and parasite water column abundance both were significantly related to temperature, but the responses lagged 1 month behind temperature. Parasite water column abundance was the highest during August (up to 1,200 cells/l) and was significantly related to P. marinus weighted prevalence in wild oysters, and to wild oyster mortality suggesting that parasites are released in the environment via both moribund and live hosts (i.e. through feces). Incidence was not significantly related to parasite water column abundance, which seems to indicate the absence of a linear relationship or that infection acquisition is controlled by a more complex set of parameters. PMID- 16476184 TI - A murine model for cerebral toxocariasis: characterization of host susceptibility and behaviour. AB - Toxocara canis, the parasitic roundworm of dogs, can infect a number of paratenic hosts, such as mice and humans, due to the widespread dissemination of its ova in the environment. In these paratenic hosts, larvae have been shown to exhibit a predilection for the central nervous system, resulting in an increasing number of parasites migrating to the brain as infection progresses. In an initial experiment, we investigated the differential brain involvement of T. canis in 7 strains of inbred mice, and chose 2 strains, susceptible (BALB/c) and resistant (NIH) to cerebral infection. In a second experiment, both strains were investigated in terms of course of migration, larval accumulation, and behavioural response to T. canis infection. Results revealed that infected BALB/c mice took significantly longer to drink from a water source (following a period of deprivation), compared with control mice, indicating some degree of memory impairment. Cerebral larval recoveries from both strains of mice demonstrated variation between the two experiments, suggesting that larval burdens may not be a reliable indicator of susceptibility or resistance to T. canis infection. The percentage of total recovered larvae in each organ may be a better representation of larval distribution. Our model system may provide insights into the impact of chronic geohelminth infection on cognitive development. PMID- 16476185 TI - Effectiveness of cognitive-behavioural, person-centred and psychodynamic therapies as practised in UK National Health Service settings. AB - BACKGROUND: Psychotherapy's equivalence paradox is that treatments have equivalently positive outcomes despite non-equivalent theories and techniques. We compared the outcomes of contrasting approaches practised in routine care. METHOD: Patients (n = 1309) who received cognitive-behavioural therapy (CBT), person-centred therapy (PCT) and psychodynamic therapy (PDT) at one of 58 National Health Service (NHS) primary and secondary care sites in the UK during a 3-year period completed the Clinical Outcomes in Routine Evaluation-Outcome Measure (CORE-OM) at the beginning and end of their treatment. Therapists indicated which treatment approaches were used on an End of Therapy form. We compared outcomes of six groups: three treated with CBT, PCT or PDT only, and three treated with one of these plus one additional approach (e.g. integrative, supportive, art), designated CBT+1, PCT+1 or PDT+1 respectively. RESULTS: All six groups averaged marked improvement (pre-post effect size = 1.36). Treatment approach and degree of purity ('only' vs. '+1') each accounted for statistically significant but comparatively tiny proportions of the variance in CORE-OM scores (respectively 1% and 0.5% as much as pre-post change). Distributions of change scores were largely overlapping. CONCLUSIONS: Results for these three treatment approaches as practised routinely across a range of NHS settings were generally consistent with previous findings that theoretically different approaches tend to have equivalent outcomes. Caution is warranted because of limited treatment specification, non-random assignment, lack of a control group, missing data and other issues. PMID- 16476186 TI - A comparison of the effects on bispectral index of mild vs. moderate hypothermia during cardiopulmonary bypass. AB - BACKGROUND AND OBJECTIVE: Bispectral Index correlates with depth of hypnosis in adult patients. Bispectral index monitoring has been separately examined and validated during both mild and moderate hypothermic cardiopulmonary bypass. However, the effect on bispectral index of these two differing cardiopulmonary bypass temperature regimes has not previously been compared, particularly with the use of volatile anaesthetic agents. METHODS: Following Institutional Ethics Committee approval and informed consent, 30 patients undergoing coronary artery bypass grafting had additional bispectral index monitoring. Sixteen patients had mild hypothermic bypass (32-34 degrees C), and 14 had moderate hypothermic bypass (28-30 degrees C). Anaesthesia was standardized, and was maintained with enflurane throughout surgery. Scores were recorded at 14 specific event-related time points during surgery. RESULTS: This study has demonstrated that in this adult population, a relationship exists between bispectral index score and temperature throughout the duration of cardiac surgery, which included mild or moderate hypothermic cardiopulmonary bypass. It also demonstrated that patients which were categorized as having received moderately hypothermic cardiopulmonary bypass had lower bispectral index scores than those patients who received mild hypothermic cardiopulmonary bypass, at time points corresponding approximately with median steady-state bypass temperature, pre-re-warming and early re-warming periods. CONCLUSIONS: Moderate hypothermic cardiopulmonary bypass has lower bispectral index scores when compared to mild hypothermic cardiopulmonary bypass, at a steady state of inspired enflurane. This may reflect an actual increase in the depth of hypnosis. However, as this study did not address the physico chemical changes of the volatile agent or the accuracy of the bispectral index algorithm at lower temperatures, we would not advocate a change in practice based solely on this report. PMID- 16476187 TI - Changing paradigms in the immunologic science of allergy. PMID- 16476188 TI - Vaccination and anaphylaxis. AB - The incidence of anaphylactic or severe allergic reactions to vaccines is very low, less than one case per million vaccine doses. Larger studies from later years report no deaths. The cause of the reaction is usually not the immunizing antigen itself, but rather some other vaccine ingredient such as egg protein from the production process or gelatin added as a stabilizer. Most people with egg allergy can be vaccinated without any reaction. Vasovagal reactions with or without hyperventilation are common after vaccination. They can be rather dramatic and are often mistaken for anaphylactic reactions. Correct diagnosis is important in making it possible to vaccinate those who might otherwise run the risk of serious infections. PMID- 16476189 TI - Cardiac anaphylaxis: pathophysiology and therapeutic perspectives. AB - Cardiac anaphylaxis refers to the functional and metabolic changes in the heart caused by the anaphylactic release of histamine and vasoactive products of arachidonic acid cascade by mast cells and basophils. As in most type I hypersensitivity-based diseases, histamine plays a key role in the pathophysiology of cardiac anaphylaxis. In the heart, mast cell activation and histamine release are controlled by multiple endogenous mechanisms, including adrenergic neural control, histamine-dependent negative feedback operated through H2 receptors, and the endogenous generation of nitric oxide (NO) and carbon monoxide (CO). All these mechanisms can be targeted by substances that have revealed a clear-cut effect in blunting cardiac anaphylaxis in experimental animal models, and could be developed as potential, novel anti-anaphylactic drugs. In this article, we discuss new findings and significant trends related to this topic. PMID- 16476190 TI - The role of T cells in drug reaction. AB - T cells are major protagonists of immune-mediated adverse drug eruptions. Immunohistochemical observation as well as isolation and functional characterization of T cells infiltrating the affected tissues provided new insights into the pathomechanisms of the diverse clinical manifestations of drug hypersensitivity, and permitted the recognition of T cell-mediated cytotoxicity against drug-loaded resident cells as a major mechanism of tissue damage. PMID- 16476191 TI - Indoleamine-2,3-dioxygenase modulation of allergic immune responses. AB - Induction of immunologic tolerance is highly desirable in the treatment and prevention of allergy and other immune disease states in which the immune response to foreign or self antigens has become overactive. Indoleamine-2,3 dioxygenase (IDO), an enzyme classically known for its role in the tryptophan degradation pathway, has recently emerged as an important immunomodulator of T cell function and inducer of tolerance. The induced expression of IDO by dendritic cells may suppress T-cell responses and promote tolerance either through direct effects on T cells (mediated by tryptophan depletion or tryptophan metabolites) or through effects of IDO on the dendritic cell. In addition to the potential role of IDO in promoting tolerance in pregnancy, transplantation, and autoimmunity, its role in modulating allergic responses has more recently been investigated, raising the possibility that IDO and its metabolites may be novel targets for immunomodulation in allergy and asthma. PMID- 16476192 TI - Suppressor of cytokine signaling 3 (SOCS3) in Th2 cells evokes Th2 cytokines, IgE, and eosinophilia. AB - Atopic dermatitis, allergic rhinitis, and bronchial asthma are allergic immune disorders characterized by a predominance of T helper 2 (Th2) cells, the resulting elevation of allergen-specific immunoglobulin E (IgE), and mast cell- and eosinophil-associated inflammation. The cytokine environment at the site of the initial antigen stimulation determines the direction of helper T-cell differentiation into Th1 or Th2 cells. Therefore, negative regulators of cytokine signaling, suppressors of cytokine signaling (SOCS) proteins, play an important role in Th2-mediated allergic responses through the control of the balance between Th1 and Th2 cells. SOCS3 and SOCS5 are predominantly expressed in Th2 and Th1 cells, respectively, and they reciprocally inhibit the Th1 and Th2 differentiation processes. In this article, we discuss the role of SOCS3 and SOCS5 proteins in atopic asthma and allergic conjunctivitis and explore the potential of SOCS proteins as targets for therapeutic strategies in allergic disorders. PMID- 16476193 TI - Selective roles and dysregulation of interleukin-10 in allergic disease. AB - There is mounting evidence to support a role for interleukin (IL)-10 in the regulation of both allergic and nonallergic immune responses. The current view is that IL-10 influences Th2-driven allergic processes by altering the interplay between Th1 and Th2 effector cells. However, lack of a clear delineation of the different types of IL-10-secreting regulatory T cells poses a major challenge in defining IL-10-mediated immune pathways, which govern the development, persistence, and modulation of allergic status. This is compounded by observations that undermine the credibility of the Th1/Th2 paradigm as a model for allergic disease. Nevertheless, enhanced IL-10 secretion by T cells during conventional immunotherapy, coupled with evidence of a link between genetics and high IL-10 production to a specific allergen, suggests that IL-10 induction is an appropriate goal of therapy. Knowledge of the targeted patient population and design of an immunogen (ie, peptide or modified allergen) within this context are likely to provide improved results over conventional immunotherapy. PMID- 16476194 TI - Airway acidification: interactions with nitrogen oxides and airway inflammation. AB - Airway acidification is increasingly appreciated to occur in inflammatory obstructive airway diseases, resulting from acid reflux and aspiration and from direct acid formation in the airways. Acidity activates oxidants and nitrogen oxides to create a potent antimicrobial environment. Neurogenic inflammation is triggered by airway or esophageal acidification, innate immune cells are affected by acidity, and there are pathways by which the acquired immune system also can be activated by the chemistry of an acidic airway. Measuring airway acidity is now readily achievable with noninvasive breath assays, a procedure that has opened a window on the need to understand airway pH homeostasis in health and pH dysregulation in disease. PMID- 16476197 TI - Genetics of food allergy. AB - Allergic reactions to foods are an important medical problem throughout the industrialized world. The occurrence of food allergy appears to be strongly influenced by genetics, but the basis of the genetic predisposition to food allergy has not been differentiated from that for atopy in general. In addition, genetic susceptibility alone does not explain the prevalence of food allergy satisfactorily, leaving ample room to consider the importance of environmental influences (external, maternal, and gastrointestinal environment) and interactions between the host and the environment. Several features of food allergy are highlighted in this review: 1) patients with severe food allergies are overwhelmingly atopic, but food allergy occurs only in approximately 10% of patients with other atopic diseases; 2) most patients are clinically reactive to a single food, and although a substantial minority have multiple food allergies, the variety of bone fide food allergies in a given individual is limited; 3) foods contain multiple proteins whereas only a small subset are allergenic; 4) there is likely an important contribution of the environment, becoming manifest in genetically susceptible individuals. PMID- 16476199 TI - Maternal allergen exposure as a risk factor for childhood asthma. AB - The role of early allergen exposure has been a central focus in the effort to curtail the rising rates of allergic disease. Exposures in pregnancy have been of interest because the first signs of disease are often seen in early infancy. However, so far, strategies to avoid or reduce allergen exposure in pregnancy have had disappointing results in reducing disease. Although there is growing evidence that the fetus is exposed to allergens, this appears to be physiologic and not a risk factor for allergic disease. It is still not clear if this exposure has some role in the context of normal tolerance. Although infants who later develop allergic disease show some differences in neonatal allergen specific responses, these are not consistent. Furthermore, the magnitude and frequency of responses do not correlate well with maternal allergen exposure, and it now seems likely that these responses do not reflect true immunologic "memory." Despite this, there is accumulating evidence that a number of other exposures in pregnancy have the capacity to significantly influence immune development and alter the risk for allergic responses to allergens. This underscores the importance of further defining the events in this early period, to determine causal pathways and better strategies for prevention. PMID- 16476198 TI - Eosinophil Function in Eosinophil-associated Gastrointestinal Disorders. AB - Eosinophil-associated gastrointestinal disorders (EGIDs) are characterized by a rich eosinophilic inflammation of the gastrointestinal tract in the absence of known causes for eosinophilia or other gastrointestinal disorders. These disorders include eosinophilic esophagitis, eosinophilic gastritis, eosinophilic gastroenteritis, eosinophilic enteritis, and eosinophilic colitis, and are being recognized with increasing frequency. Clinical studies suggest that eosinophils have a pathogenic role in EGID; however, the function of eosinophils in these disorders remains an enigma. In this review, we briefly describe the clinical features of EGID of the esophagus, small bowel, and colon. We summarize recent experimental analysis examining the underlying contribution of eosinophils to disease pathogenesis and discuss possible therapeutic approaches for the treatment of these diverse diseases. PMID- 16476200 TI - The role of defensins in virus-induced asthma. AB - Respiratory viruses appear to play a central role in asthma pathogenesis. In addition to a major role in triggering asthma exacerbations, viral infections early in life may play a role in disease development. Although defensins were initially identified as antimicrobial peptides, recent studies have demonstrated that they have a much broader range of actions that are of relevance to virally induced asthma. Defensins are not only induced during viral infections, they exert direct antiviral actions against some enveloped viruses, and also can impair viral infection of cells. In addition, defensins modulate the activity of several cell types that contribute to innate immunity, including mast cells, epithelial cells, natural killer cells, and dendritic cells. They also have the capacity to modulate adaptive immunity and to contribute to airway remodeling. In this article, recent advances in these areas are reviewed in the context of the potential role of defensins in virus-induced asthma. PMID- 16476201 TI - Effects of bovine oviduct epithelial cells, fetal calf serum and bovine serum albumin on gene expression in single bovine embryos produced in the synthetic oviduct fluid culture system. AB - In this study the synthetic oviduct fluid (SOF) system with bovine oviduct epithelial cell (BOEC) co-culture is compared with an SOF system with common protein supplements. One thousand six hundred bovine embryos were cultured in SOF media supplemented with BOEC, fetal calf serum (FCS) and bovine serum albumin (BSA). Eight different culture groups were assigned according to the different supplementation factors. Developmental competence and the expression levels of five genes, namely glucose transporter-1 (Glut-1), heat shock protein 70 (HSP), connexin43 (Cx43), (2)-actin (ACTB) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), analysed as mRNA by using reverse transcription-polymerase chain reaction, were measured on bovine embryos cultured for 9 days. Gene expression of these in vitro-produced embryos was compared with the gene expression of in vivo produced embryos. There was no significant difference found in embryo developmental competence between the Day 9 embryos in BOEC co-culture, FCS and BSA supplements in SOF media. However, differences in gene expression were observed. With respect to gene expression in in vivo and in vitro embryos, BOEC co-culture affected the same genes as did supplementation with FCS and BSA. HSP was the only gene that differed significantly between in vitro and in vivo embryos. When the different in vitro groups were compared, a significant difference between the BOEC co-culture and the FCS supplementation groups due to Glut-1 expression was observed. PMID- 16476202 TI - Reproductive effects after exposure of male mice to vincristine and to a combination of X-rays and vincristine. AB - This study was performed to investigate the effects of exposure to vincristine (VCR) alone (1 mg kg(-1) or 2 mg kg(-1)) and in combination with X-rays (0.25 Gy + 1 mg kg(-1) VCR or 1.00 Gy + 2 mg kg(-1) VCR) on the quality and quantity of spermatozoa, and the offspring of exposed Pzh : Sfis male mice. Both VCR and X rays plus VCR reduced testis weight and sperm count, caused deterioration with respect to sperm morphology, and caused a slight increase in DNA damage. Exposure of some stages of male germ cells to high doses of VCR either alone or in combination with X-rays reduced the rate of pregnant females and the fertility of males. Such treatments reduced the number of total and live implantations, and induced dominant lethal mutations. The results of this study demonstrated the reproductive genotoxicity of VCR with or without X-rays, but did not unequivocally confirm their ability to cause external malformations in offspring. PMID- 16476203 TI - Luteal stage dependence of pituitary response to gonadotrophin-releasing hormone in cyclic dairy ewes subjected to synchronisation of ovulation. AB - Possible hormonal aberrations precluding conception or maintenance of pregnancy in dairy ewes subjected to ovulation synchronisation were investigated in this study. The pituitary response to exogenous gonadotrophin-releasing hormone (GnRH) was tested at different luteal stages in 36 ewes. Oestruses were synchronised by using progestagen-impregnated sponges and the animals were randomly allotted into one of three treatment groups (A, B and C; n = 12 for each). Treatments commenced on Days 4, 9 and 14 of the new cycle (oestrus was defined as Day 0). Ewes were given two GnRH injections, 5 days before and 36 h after a prostaglandin F2+/- (PGF2+/-) injection, and the animals were inseminated 12-14 h after the second GnRH injection (modified OVSYNCH). For luteinising hormone (LH) determination blood samples were withdrawn from six ewes of each group at the time of GnRH administration, and 30, 90, 180, 270 and 360 min later. Progesterone was assayed in samples taken every other day starting from oestrus and for 17 days after the second GnRH injection, and in an additional sample collected on the day of insemination. After the first GnRH injection, the LH concentration was higher in Group C than in Groups B and A (mean +/- s.d.: 64.8 +/- 10.0 ng mL(-1), 41.3 +/- 3.7 ng mL(-1) and 24.6 +/- 9.0 ng mL(-1), respectively; P < 0.05), whereas after the second GnRH injection a uniform LH release was found in all groups. PGF2+/- caused a significant decrease in progesterone (P4) concentration in all groups; however, at artificial insemination ewes that conceived had significantly lower P4 concentration in comparison with those that failed to conceive. As early as Day 5, pregnant animals had higher P4 concentrations than non-pregnant animals. Overall, 21 animals conceived (seven, nine and five ewes from Groups A, B and C, respectively). These results indicate that the proposed protocol is equally effective in inducing a preovulatory LH surge at any stage of the luteal phase, and that elevated P4 concentration along with a delayed P4 increase should be considered as a causative factor for inability to conceive. PMID- 16476204 TI - In vitro and in vivo culture effects on mRNA expression of genes involved in metabolism and apoptosis in bovine embryos. AB - Bovine blastocysts produced in vitro differ substantially from their in vivo derived counterparts with regard to glucose metabolism, level of apoptosis and mRNA expression patterns. Maternal embryonic genomic transition is a critical period in which these changes could be induced. The goals of the present study were twofold: (1) to identify the critical period of culture during which the differences in expression of gene transcripts involved in glucose metabolism are induced; and (2) to identify gene transcripts involved in apoptosis that are differentially expressed in in vitro- and in vivo-produced blastocysts. Relative abundances of transcripts for the glucose transporters Glut-1, Glut-3, Glut-4 and Glut-8, and transcripts involved in the apoptotic cascade, including BAX, BCL-XL, XIAP and HSP 70.1, were analysed by a semiquantitative reverse transcription polymerase chain reaction assay in single blastocysts produced in vitro or in vivo for specific time intervals, that is, before or after maternal embryonic transition. Whether the culture environment was in vitro or in vivo affected the expression of glucose transporter transcripts Glut-3, Glut-4 and Glut-8. However, the critical period during culture responsible for these changes, before or after maternal embryonic transition, could not be determined. With the exception of XIAP, no effects of culture system on the mRNA expression patterns of BAX, BCL-XL and HSP 70.1 could be observed. These data show that expression of XIAP transcripts in expanded blastocysts is affected by in vitro culture. These findings add to the list of bovine genes aberrantly expressed in culture conditions, but do not support the hypothesis that maternal embryonic transition is critical in inducing the aberrations in gene expression patterns studied here. PMID- 16476205 TI - Early onset of embryonic mortality in sub-fertile families of rainbow trout (Oncorhynchus mykiss). AB - Survival during early embryonic development is highly variable in oviparous fishes and appears to be related to events associated with the female at the time of ovulation and spawning. The goal of this study was to identify critical periods of mortality associated with early embryonic development in egg batches from female rainbow trout (Oncorhynchus mykiss) that were checked for ovulation every 5-7 days. The experiment was designed to specifically remove post-ovulatory ageing and reduce paternal variability. Embryo viability in 269 single-pair-mated families was systematically tracked at the following five stages: second cleavage (0.5 days post fertilisation (dpf)), elevated blastula (2.5 dpf), embryonic shield (6 dpf), embryonic keel (9 dpf), and retinal pigmentation (19 dpf). At each of the five stages families with embryo viability assessments of <80% were classed as sub-fertile, whereas those with >80% embryo viability were classed as fertile. Embryo viability in sub-fertile families was distinctly reduced at 0.5 dpf, in contrast to fertile families, but remained constant from that point through to 19 dpf. These results suggest that the critical period of early embryonic mortality in sub-fertile families of rainbow trout parallels events that occur at or shortly after fertilisation and is independent of post-ovulatory aging. PMID- 16476206 TI - Highly efficient and reliable chemically assisted enucleation method for handmade cloning in cattle. AB - The purpose of the present study was to find an efficient and reliable chemically assisted procedure for enucleation related to the handmade cloning (HMC) technique. After in vitro maturation oocytes were incubated in 0.5 microg mL(-1) demecolcine for 2 h. Subsequently, zonae pellucidae were digested with pronase, and one-third of the cytoplasm connected to an extrusion cone was removed by hand using a microblade. The remaining two-thirds were used as recipients for HMC, and reconstructed and activated embryos were cultured for 7 days. The time-dependent manner of the development of extrusion cones, the efficiency (oriented bisection per oocyte; 94%), reliability (success per attempted enucleation; 98%), and the blastocyst per reconstructed embryo rates (48%) were measured. Ultrastructural analyses demonstrated that demecolcine treatment resulted in disoriented and haphazardly orientated microtubules. The general ultrastructure of the oocyte organelles, however, appeared to be unaltered by the treatments. Considering that no oocyte selection based on polar body presence was performed, this system seems to be more efficient and reliable than any other enucleation method. Moreover, expensive equipment (inverted fluorescence microscope) and a potentially harmful step (staining and ultraviolet illumination) can be eliminated from the HMC procedure without compromising the high in vitro efficiency. PMID- 16476207 TI - Stereomicroscopic and histological examination of bovine embryos following extended in vitro culture. AB - Attempts to support survival of mammalian embryos after hatching have met with limited success, although some mouse studies have reported growth at the post implantation stage. The aim of the present research was to establish and characterise an in vitro culture system that could support extended growth and differentiation of bovine embryos. Abattoir-derived oocytes were matured and fertilised in vitro. Presumptive zygotes were cultured in modified synthetic oviduct fluid (SOFaaci) medium supplemented with 5% cow serum (CS). On Day 9, single hatched blastocysts (n = 160) were randomly allocated to SOFaaci supplemented with either 5% bovine serum albumin, 5% CS, 5% fetal calf serum (FCS) or SOF only and cultured on a collagen gel substrate for up to 45 days. Embryos were evaluated at various time-points until complete disaggregation or the total disappearance of embryonic cells. Blastocyst viability post hatching was severely compromised in protein-free SOFaaci medium. Addition of FCS generated increased embryonic growth for the longest time period (Day 45) when compared to the other groups. Long-term survival of embryonic cells was observed stereomicroscopically by the proliferation and development of three-dimensional tubular structures to 85% confluence in culture. Haematoxylin and eosin staining of morphological structures obtained from all treatment groups revealed embryos displaying trophoblast, inner cell mass and hypoblast development to varying degrees. Regardless of treatment, extended in vitro culture did not result in development comparable with that described for in vivo embryos. In the present work, however, there was evidence of extended culture of bovine embryos beyond that achieved previously. However, further research is required to identify the exact requirements for extended in vitro culture for bovine embryos. PMID- 16476208 TI - DNA methylation patterns in in vitro-fertilised goat zygotes. AB - Recent studies have shown that zygote demethylation patterns in mammals are variable among species. However, the methylation patterns of goat zygotes have not yet been reported on. In the present study, using immunofluorescence staining with an antibody against 5-methylcytosine, the methylation patterns of in vitro derived goat zygotes were studied. The results indicate that goat zygotes do not undergo active global demethylation during pronuclei development, which is similar to the situation in ovine, but not in murine or bovine zygotes. This is believed to be the first report regarding methylation in goat zygotes. PMID- 16476209 TI - Accumulation and distribution of neutral lipid droplets is non-uniform in ovine blastocysts produced in vitro in either the presence or absence of serum. AB - Sheep zygotes were cultured in serum-free or serum-supplemented media to determine effects on blastocyst yields and within-blastocyst abundance and distribution of neutral lipid droplets. Embryos cultured in synthetic oviduct fluid supplemented with bovine serum albumin (0.4% w/v) (SBSA) generated similar blastocyst yields (mean +/- s.e.m. = 20% +/- 5) to those in synthetic oviduct fluid supplemented with serum (10% v/v) from ewes fed a diet containing 0% (SZFO; 26% +/- 2) or 3% fish oil (S3FO; 23% +/- 3). SBSA zygotes generated more good quality blastocysts than their SZFO or S3FO counterparts (P < 0.05). Within blastocyst abundance of neutral lipid droplets was non-uniform; data were collected from discrete embryo sectors (each = 2700 microm2) representing highest (H), intermediate (I) and lowest (L) densities of accumulation. For all sectors, area (microm2) occupied by lipid droplets in SBSA blastocysts (mean H = 470; I = 370; L = 245) was smaller (P < 0.01) than occupied in others (SBSA : SZFO = 1 : 1.41, 1 : 1.48 and 1 : 1.42; SBSA : S3FO = 1 : 1.36, 1 : 1.30 and 1 : 1.31; data for H, I and L, respectively). Among S3FO blastocysts only, inferior quality was associated with greater lipid abundance. Overall, embryo culture in the presence of serum increased neutral lipid droplet abundance but accumulation was non uniform. PMID- 16476212 TI - [Treatment following suicide attempt. Therapeutic guarantee needs]. PMID- 16476213 TI - [Prevention of repeated suicide attempts. Evaluation of the treatment effectiveness]. AB - INTRODUCTION: Repetition of suicide attempts occurs frequently. There is a lack of knowledge as to which treatment strategies are most effective. MATERIALS AND METHODS: STUDY 1: A cohort of 200 psychiatric inpatients, consecutively admitted for current suicide attempt, were studied as to the association among quality level of examination, treatment and referral to aftercare and repetition of attempted suicide in one year. STUDY 2: Three cohorts of psychiatric inpatients, consecutively admitted for current suicide attempt, were compared for repetition of suicide in one year. The 2001-2002 cohort of 173 inpatients had received problem-oriented therapy and been provided with a contact card to a psychiatric emergency room. The 1994-1995 control cohort of 200 inpatients had received quality-ensured standard treatment. The 1989 control cohort of 126 inpatients had received standard treatment only. Similarly, two cohorts of psychiatric emergency outpatients, referred after current suicide attempts, were compared: 116 patients in 2001-2002 and 75 patients in 1994-1995. RESULTS: A higher quality level of examination and treatment was significantly related to lack of repetition of attempted suicide in one year. No association was found between higher quality level of referrals to aftercare and no repetition of attempted suicide. The rate of repetition of suicide attempts in one year was reduced from 30% to 24% after establishment of quality assurance and problem-oriented therapy, but the reduction was not significant. CONCLUSION: It is still uncertain which treatment modalities of patients admitted for attempted suicide are most effective. PMID- 16476214 TI - [Sexual dysfunction in the menopause. Incidence, pharmacological treatment and side effects]. AB - The frequency of female sexual dysfunction increases with age, and the menopause has a negative influence on sexual life. Pharmacological treatment options of female sexual dysfunctions in the menopause include hormone therapy and sildenafil. Few randomised controlled studies have been done, and there is evidence suggesting that systemic hormone therapy, such as estrogen, estrogen/progesterone, estrogen/testosterone and tibolone, has a positive impact on sexual dysfunction in the menopause. There is evidence that local estrogen relieves vaginal dryness and dyspareunia. The recent discoveries of the side effects of hormone therapy necessitate careful evaluation of the indication for hormone therapy, and the duration of treatment is recommended to be as short as possible. The long-term side effects of testosterone in women have not yet been fully investigated. Sildenafil has shown a positive effect on female sexual dysfunction only in a limited group of women: those with arousal problems without desire problems. This result demands a focus on new pharmaceutical products, and at present the effect of testosterone and selective estrogen receptor modulators on female sexual dysfunction is being investigated. PMID- 16476215 TI - [Pregnancy and recreational physical activity]. AB - Recent research indicates that physical activity before and during pregnancy helps lower the risk of gestational diabetes and preeclampsia. Physical activity during pregnancy has no negative effects on the risk of preterm birth, low birth weight and birth complications; however, thus far the topic has not been widely explored. In the light of the available findings, although of limited scope, it seems justified to encourage healthy pregnant women who are already physically active at the onset of pregnancy to maintain their activities but to reduce their intensity and to encourage pregnant women who are not already physically active to initiate physical activity. PMID- 16476216 TI - [Erythropoietin for patients with malignant disease. An analysis of a systematic Cochrane review]. PMID- 16476217 TI - [Social profile of young people seeking treatment for cannabis abuse]. AB - INTRODUCTION: The aim of this study was to determine the social profile and sociodemographic characteristics of young people seeking treatment for cannabis dependence. MATERIALS AND METHODS: Participants (n = 75) aged 18-30 who met the ICD-10 diagnostic criteria for cannabis dependence and sought treatment were interviewed about their social and psychological background. Clients meeting the diagnostic criteria for alcohol or substance dependence were excluded. RESULTS: The mean age was 24 years; 82% were male; 87% were receiving financial assistance from the state. 8% had lost a parent, 61% reported that at least one of their parents abused alcohol, cannabis or hard drugs. 52% had had previous conduct problems. 49% had been exposed to bullying. 70% had changed school two or more times, 50% had a history of learning difficulties, 44% had been expelled from school. CRIMINAL BACKGROUND: 58% had received a sentence, 20% because of violence; of their parents, 19% had received a sentence, 10% because of violence. DISCUSSION: The results indicate that the previously identified risk factors for substance abuse are also present among young people in treatment for cannabis dependence. These young people are also characterised by having changed school many times. The cannabis abusers in the present study were younger and less socioeconomically stable than the participants in the five previously published controlled trials on treatment for cannabis dependence. Development and testing of cannabis dependence interventions with a sample that is younger, less educated and less socioeconomically stable than has been studied in prior research are needed. PMID- 16476218 TI - [General practice research supervision]. AB - INTRODUCTION: General practice is a subject with a relatively short scientific tradition. The purpose of this study was to elucidate who gives long-cycle general practice research supervision in Denmark, who is supervised and how research students get on. MATERIALS AND METHODS: All research students with research advisers in the field of general practice filled out a questionnaire in 1997 (n = 50) and 2003 (n = 52). There were questions about project/research training, professional education, advisers and the students' attitude to taking on advisory functions in the future. In 2003, 48 of those from the 1997 cohort also answered a follow-up questionnaire. RESULTS: The number of research students with a connection to the general practice research field was fairly constant from 1997 to 2003. The number of permanently employed general practice advisers, on the other hand, has doubled, and these now undertake more of the advisory functions. The great majority of research students complete their projects, most of them at PhD level. Most of the research students surveyed indicated that they are prepared to take on advisory functions when they have acquired the competence to do so. DISCUSSION: The general practice research students vary greatly with regard to age, interests and career paths, but almost all complete their research studies. This may be due to the flexible framework for research in general practice and the increase in recent years of advisory capacity in general practice. This positive development is expected to continue through the development of networks, course activities and continuing follow-up of the advisory functions. PMID- 16476219 TI - [Extralegal coercive measures in psychiatric wards-- patients' perspectives]. AB - INTRODUCTION: A revision of the Danish Mental Health Act is planned to take place in 2005-2006. The Ministry of Health requires investigation into whether extralegal coercive measures take place in psychiatric wards. Extralegal coercion refers to restrictions and rules not included in the current Mental Health Act and considered by patients to be coercion. MATERIALS AND METHODS: Twenty-two psychiatric patients participated in four group interviews directed by two physicians at four University Hospitals in Copenhagen, Denmark. The patients were asked to define what they perceived as extralegal coercion, house rules and psychiatric treatment. RESULTS: The majority of patients perceived lack of single rooms, restrictions on leaving the ward and shielding, i.e., being confined to one's room, to be extralegal coercion. A few also found the pressure to take medication and collection of clothes items to be extralegal coercion. Rules on smoking, clothing, day and night routines, the use of radio and TV, drugs, alcohol, visits, eating and the use of telephones were considered not extralegal coercion but necessary house rules. House rules should be clear and stated in writing. Restrictions on the use of money and locking of the front door at night were accepted in certain situations, hence not considered to be extralegal coercion. DISCUSSION: Extralegal coercion does occur in psychiatric wards but in certain situations is accepted if documented. House rules are considered to be necessary and not extralegal coercion. Increased use of compulsory protocols for documenting coercive measures was not considered to be necessary, but documentation in patients' files was useful alternative. PMID- 16476220 TI - [Metabolic control by means of insulin in patients with type 2 diabetes and acute myocardial infarction (DIGAMI 2): effects on mortality and morbidity--secondary publication]. AB - Patients with diabetes have an unfavourable prognosis after an acute myocardial infarction. The DIGAMI 2 study investigated the effect of various metabolic treatment strategies in type 2 diabetic patients with acute myocardial infarction: acutely introduced, long-term insulin treatment did not improve survival when compared with conventional management at similar levels of glucose control. However, good glucose control seems important since the glucose level was found to be a strong predictor of long-term mortality in this patient category. PMID- 16476221 TI - [Risk adjustment for surgery of congenital heart disease--secondary publication]. AB - Risk adjustment for specialties covering many diagnoses is difficult. The Risk Adjusted classification for Congenital Heart Surgery (RACHS-1) was created to compare the in-hospital mortality rate of groups of children undergoing surgery for congenital heart disease. We applied the classification to the operations performed at Skejby Sygehus (1996-2002) and found that RACHS-1 can be used to predict the in-hospital mortality rate and length of stay in the intensive care unit in a Danish center for congenital heart surgery. The mortality rate was similar to that reported by larger centers. PMID- 16476222 TI - [Self-reported stress and risk of breast cancer--a secondary publication]. AB - A possible association between stress and risk of breast cancer has been assessed in different study designs with conflicting results. We prospectively followed 6,689 women from the Copenhagen City Heart Study for 18 years for a first-time diagnosis of primary breast cancer. The women were asked about stress intensity and frequency at baseline in 1981-83. Women reporting high levels of stress were at lower risk of breast cancer (hazard ratio = 0.60; 95% CI: 0.37-0.97) than women reporting low levels of stress. This association was most pronounced among women who received hormone therapy. PMID- 16476223 TI - [Acute multifocal osteomyelitis caused by haemolytic group B streptococci]. AB - Invasive infections due to group B streptococci (Streptococcus agalactiae) usually occur in the peri- and neonatal setting or in adults with chronic underlying diseases. In the presented case a severe infection of haematogenous osteomyelitis caused by group B streptococci in a previously healthy 68-year-old woman is reported. Group B streptococci were isolated in three of three blood culture bottles drawn from the patient at the time of admission to hospital due to fever of unknown cause and pain in the upper and lower extremities as well as the back. A technetium scan showed involvement of the left shoulder, the left sacroilial joint, Th-9, Th-12 and L5. In addition, there was increased activity in the left foot, the right side of the mandibula and probably both hands (insufficiently scanned). The patient was successfully treated with penicillin G and subsequently with oral penicillin V for a total of 12 weeks. PMID- 16476224 TI - [Refractory anaemia successfully treated with thalidomide]. AB - A 63-year-old male was admitted to our department in 2001 because of autoimmune haemolytic anaemia and splenomegaly. Bone marrow examination revealed an erythroid hyperplasia and a normal chromosome analysis. Splenectomy was performed in 2002, resulting in severe transfusion-dependent anaemia with a need for two blood transfusions each week. Treatment with prednisolone, erythropoietin and danocrine was without effect. A new bone marrow examination then showed myelodysplastic syndrome. Treatment with thalidomide, 50-150 mg/d, and prednisolone, 25 mg/d, resulted in an increase in haemoglobin to 8.9 mmol/l during the following months. The use of immunomodulatory and anti-angiogenetic agents in refractory anaemia needs more study. PMID- 16476228 TI - [Lumbar puncture 2]. PMID- 16476229 TI - [Lumbar puncture 1]. PMID- 16476230 TI - [Lumbar puncture 3]. PMID- 16476231 TI - [Lumbar puncture--laugh or cry?4]. PMID- 16476233 TI - [PET or SPECT for postsynaptic nerve cells in the striatum can possibly contribute to better insight to manganese-induced parkinsonism]. PMID- 16476235 TI - [Physicians are amateurs when it comes to medical history--thanks for the compliment!]. PMID- 16476240 TI - [CT of the thorax in colorectal cancer staging 2]. PMID- 16476238 TI - [Treatment of cervix cancer with radical hysterectomy is a task for district departments]. PMID- 16476253 TI - [Pemphigus]. AB - Pemphigus is an infrequent, organ-specific, autoimmune bullous disease, which affects the skin, mucous membranes and appendages. Histopathologically, it is characterized by acantholysis. Pemphigus has classically been divided into two major groups, pemphigus vulgaris and pemphigus foliaceus, with their respective clinical variants pemphigus vegetans and pemphigus erythematosus. In recent years, new variants of pemphigus have been described: paraneoplastic pemphigus, IgA pemphigus and pemphigus herpetiformis. This article reviews the epidemiology, etiopathogenesis, clinical symptoms, diagnosis, treatment and prognosis of pemphigus. Advances in molecular biology techniques have made it possible to more precisely identify the different antigens against which antibodies are directed, and to fine-tune ELISA diagnostic techniques. Treating pemphigus vulgaris and foliaceus with general steroids has modified their prognosis; it is estimated that mortality in recent decades is less than 10 %. Managing the clinical complications that appear during the evolution of the pemphigus has contributed to reducing morbidity and mortality. PMID- 16476254 TI - [Mucous membrane pemphigoid: clinical manifestations and treatment with corticosteroids, dapsone and cyclophosphamide in 5 patients]. AB - INTRODUCTION: Cicatricial pemphigoid includes several processes which are characterized by the presence of subepidermal bullae, and which affect the mucous membranes and, more rarely, the skin. At present, the term mucous membrane pemphigoid (MMP) is more accepted than other names used in the past, as they do not clearly define the broad spectrum presented by this disease. MMP can cause significant dysfunctions, primarily in the mucous membranes. Therefore, it is necessary to diagnose the disease as soon as possible, in order to quickly initiate systemic immunosuppressive treatment. MATERIAL AND METHODS: We present our experience with 5 patients with MMP. We analyze the clinical manifestations and the response to immunosuppressive treatment during the evolution of the disease. RESULTS: The patients were aged 41 to 69 years. The most frequent location of the lesions was the oral mucosa (80 %) and the ocular mucosa (80 %), followed by the pharyngeal mucosa (60 %), laryngeal mucosa (40 %), skin, anal mucosa (20 %) and genital mucosa (20 %). Three patients received systemic corticosteroids, dapsone and cyclophosphamide, and several sessions of plasmapheresis were also associated in one patient. One patient was controlled with topical corticosteroids and dapsone. CONCLUSIONS: Many patients with MMP can present with severe secondary complications. For this reason, the diagnosis must be confirmed quickly and the appropriate treatment started as soon as possible. The association of corticosteroids, dapsone and cyclophosphamide is a combination that gives very good results. PMID- 16476255 TI - [Mucous membrane pemphigoid: IgG and IgA antibodies against the BP180 antigen]. AB - INTRODUCTION: Mucous membrane pemphigoid is a group of autoimmune bullous diseases, mediated by autoantibodies directed against different proteins in the dermoepidermal junction, including the BP180 antigen. PATIENTS, MATERIAL AND METHODS: We included five patients with MMP in this study. We studied the presence of circulating autoantibodies against the BP180 antigen and against recombinant extracellular fragments of this protein. RESULTS: We detected the presence of circulating antibodies against BP180 in all of the patients. Indirect immunofluorescence (IIF) studies were positive in 2 patients (20 %), as well as in 2 patients via salt-split studies. We found reactivity to the extracellular fragment of BP180 (LAD-1) in 3 patients, 2 of them via IgA and 1 with IgG. The serum of only 2 patients recognized the NC16A fragment, and 4 of the 5 patients had antibodies against the carboxy-terminal domain BP180 4575. CONCLUSIONS: Molecular biology techniques are very important to complement the diagnosis of MMP, especially when the results of hematoxylin-eosin or IF studies are not satisfactory for a diagnosis of MMP. PMID- 16476256 TI - [Comparative study of the treatment of psoriasis plaques with PUVA baths and narrow-band UVA (311 nm)]. AB - INTRODUCTION: Studies carried out to date comparing treatment with PUVA baths and narrow-band UVB (NBUVB) in psoriasis show that better results are obtained with NBUVB. Certain features of the protocols may interfere with the results. MATERIAL AND METHODS: Prospective study in a psoriasis unit, with treatment assigned according to availability. Of 167 patients with psoriasis plaques, 32 received PUVA baths (photosensitizer 8-methoxypsoralen at a concentration of 2.6 mg/l) and 135 received NBUVB radiation. A clearing rate of over 70 % was considered a good response. The data were described and compared between the two groups, including multivariate analysis techniques, in order to statistically control the effects of gender, number of sessions necessary for success, minimum phototoxic dose and minimum erythema dose. RESULTS: A good response was obtained in 87.5 % of the cases with PUVA baths (95 % CI: 71.0-96.5) and in 87.4 % of the cases with NBUVB (95 % CI: 80.6-92.5). No significant differences were found in the success and abandonment percentages. Among the patients who responded to the treatment, no differences were found in the number of sessions or in the cumulative dose. Gender, cumulative dose or minimum phototoxic dose and minimum erythema dose values were not associated with the response to the treatments either. CONCLUSIONS: With psoriasis plaques, similar response percentages can be expected with both treatments, using the proposed protocols. The response is regardless of gender, cumulative dose and minimum phototoxic dose and minimum erythema dose values. PMID- 16476257 TI - [50-year case of lupus vulgaris]. AB - Lupus vulgaris is the most frequent form of cutaneous tuberculosis in industrialized countries. It is a chronic and benign form of cutaneous tuberculosis that usually occurs in patients previously sensitized to Mycobacterium tuberculosis. The histopathological study shows tuberculoid granulomas that usually contain Langhans-type giant cells. Caseous necrosis is not normally found. The culture is negative in most patients. On the other hand, the Mantoux test is usually highly positive. We present the case of a 58-year-old male who developed lupus vulgaris on the left cheek over a nine-year period, and who had another similar lesion on the edge of a residual scar on the left forearm from a probable scrofuloderma suffered during childhood. PMID- 16476258 TI - [Subcutaneous sarcoidosis as the first manifestation of systemic disease]. AB - There are dermatological symptoms in up to 25% of patients with sarcoidosis, and the appearance of specific subcutaneous nodules as a manifestation of this entity is rare. They may even predate other manifestations of sarcoidosis. We present the case of a 38-year-old woman with asymptomatic subcutaneous nodules in the limbs, which corresponded to deep sarcoid granulomas in the histological study. She did not present with any extracutaneous indications. The imaging tests performed revealed right paratracheal adenopathies. This led to the diagnosis of sarcoidosis, in both its subcutaneous and pulmonary forms (stage I). Subcutaneous sarcoidosis is probably an underdiagnosed entity, as fewer than 40 cases are reflected in literature. Its value lies in the fact that it may be the first manifestation of extracutaneous or systemic sarcoidosis, which means that this form of sarcoidosis must be considered in the differential diagnosis of subcutaneous nodular lesions; close follow-up of these patients is also necessary. PMID- 16476260 TI - [Cutaneous epithelioid hemangioendothelioma]. AB - Epithelioid hemangioendothelioma is a rare vascular tumor of intermediate aggressiveness, which usually appears in adults. It generally affects soft tissues and, less frequently, the lungs and liver. Diagnosis is by histological evaluation, and the epithelioid appearance of the neoplastic endothelial cells is typical, as is the tendency to form vascular channels. Treatment is surgical excision, with broad margins. This tumor may in exceptional cases affect the skin, with few cases having been described in literature. We describe the case of a male patient with plantar epithelioid hemangioendothelioma, and we review the literature. PMID- 16476259 TI - [Acute contact eczema from paraphenylenediamine contained in temporary henna tattoos]. AB - Henna is a plant with coloring properties which is used as a dye for hair and skin, on occasion mixed with other substances. Paraphenylenediamine (PPD) is one of the additives used to accelerate drying, improve definition and darken its reddish color. There are several descriptions in literature of sensitization to the PPD contained in henna. We describe 3 new cases of allergic contact dermatitis to temporary tattoos, with skin tests positive for PPD in 2 of them. The lesions resolved in one of the patients, leaving persistent hypopigmentation. It is important for the population to be aware of this circumstance and the risk entailed by sensitization to PPD. PMID- 16476261 TI - [Spiny keratoderma of the palms: a case study]. AB - Spiny keratoderma of the palms is an infrequent entity of unknown etiology. Most of the cases described are acquired, but there are also family cases. This dermatosis is characterized by the appearance of hyperkeratotic, generally asymptomatic, projections on the palms and/or soles, measuring only a few millimeters. In the histological study, parakeratotic columns of cornoid lamellae were observed on an epidermis with a thin or absent granular layer. Its association with malignant tumor pathologies of different types has been described. We present a new case of this entity, which had the peculiarity of a relationship of the parakeratotic column with the acrosyringium in some histological sections, a finding typical of a porokeratotic eccrine ostial or dermal duct nevus. PMID- 16476262 TI - [Nonvenereal sclerosing lymphangitis of the penis: presentation of a clinical case]. AB - Nonvenereal sclerosing lymphangitis of the penis is an infrequent process that affects the distal lymphatics of this organ. It is characterized by the sudden appearance of a translucent and indurated cord on the coronal sulcus. Its etiology is unknown, although it has been related to microtraumas in the area after intense sexual activity. It is a benign, self-resolving process, so it is not necessary to perform a biopsy in early stages, and initial treatment should be conservative. We present a new case of this disease and discuss its etiopathogenic, clinical, diagnostic and therapeutic characteristics. PMID- 16476263 TI - [Poroid hidradenoma]. AB - Poroid hidradenoma is a recently described variant of eccrine poroma. This neoplasm presents structural findings of hidradenoma (solid and cystic areas) and cytological characteristics of poromas (poroid and cuticular cells, the latter showing ductal differentiation). We present a case of poroid hidradenoma in a 74 year-old woman, who consulted her physician because of a nodular lesion on the left buttock. PMID- 16476264 TI - [Scurvy: follicular purpura as a diagnostic sign]. AB - Scurvy is a set of clinical manifestations characterized by general weakness, anemia, gingivitis and cutaneous bleeding, caused by a lack of ascorbic acid in the diet. This pathology is currently a clinical rarity, although it can still be seen in cases of malnutrition associated with alcoholism or with dietary deficiencies, especially in childhood and old age. We present the case of a 45 year-old male who consulted his physician because of lower limb edema with follicular purpura, accompanied by asthenia, polyarthralgia and bleeding gums. After treatment with 1 g/day of vitamin C was initiated, the patient's symptoms quickly improved. The diagnosis of scurvy was based on the patient's clinical symptoms, dietary history and the rapid resolution of the symptoms when vitamin C supplements were initiated. PMID- 16476265 TI - [Red-violaceous linear skin lesions on upper limb]. PMID- 16476266 TI - [Agminated acquired melanocytic nevi]. PMID- 16476267 TI - [Dermatology, where are you headed?]. PMID- 16476268 TI - [Melanoma epidemiology in Spain]. AB - Spain has one of Europe's lowest melanoma incidence and mortality rates. Nevertheless, it is one of the fastest-growing pathologies in our country, with a 181.3% increase in the incidence rate in men and 205.3% increase in women. It represents 1.3% and 2.5% of malignant tumors in men and women, respectively, while the current standardized worldwide rate is respectively 2.4% and 4.9%. The highest incidence levels correspond to Tarragona for men (6.81%) and Gerona for women (8.24%), and lowest to the Canary Islands and Zaragoza (3.55% and 4.27% for women and men, respectively). This higher incidence among females differentiates Spain from the rest of Europe, where the opposite occurs. Mortality has also increased in Spain in the last few decades (1.76% in males and 1.26% in females), although this rising trend has stabilized in recent years. Spain's mortality rate is the lowest in Europe, as the EUROCARE-III study revealed. This study found an increase of 70.4% in the survival rate for men and 84.1% for women in the 1980s, while the figures for the 1990s were 73.9% for men and 89.8% for women. This low mortality rate in Spain may be due to the increase in thin melanomas, due to early diagnosis and surgical treatment. PMID- 16476269 TI - [The usefulness of isotopic lymphoscintigraphy in the study of lymphedemas]. AB - INTRODUCTION: Lymphedema is a process caused by the obstruction or destruction of the lymphatic vessels in the subcutaneous tissue. Early diagnosis and treatment can improve its clinical course, but this debilitating disease is often misdiagnosed. OBJECTIVE: To highlight the importance of isotopic lymphoscintigraphy as a simple, non-invasive technique for the assessment and diagnosis of lymphedemas. MATERIAL AND METHODS: We present 2 patients affected with genital edema, and another with edema in the right upper limb. All of these were secondary to chronic infections. An isotopic lymphoscintigraphy was performed on all three patients. RESULTS: In all three patients, the isotopic lymphoscintigraphy made it possible to confirm the diagnosis and the extent of the lymphedema. CONCLUSIONS: We believe that techniques such as phlebography, ultrasound/Doppler, CAT, magnetic resonance, cultures and biopsies should be used with all lymphedema patients in order to establish the cause of the disease. We propose that isotopic lymphoscintigraphy be added to these tests for appropriate lymphedema management. PMID- 16476270 TI - [Specific cutaneous involvement in patients with multiple myeloma. A clinicopathological, immunohistochemical and cytogenetic study of 40 cases]. AB - INTRODUCTION: Specific cutaneous involvement in patients with multiple myeloma is very rare. When it does occur, it is in patients with advanced stages of multiple myeloma and with a large tumorous mass. In this study, we analyzed 40 patients with specific cutaneous involvement from multiple myeloma, with no bony lesions underlying the skin lesions, and we reviewed the related literature. OBJECTIVES: We were particularly interested in the clinical course of these patients, including survival once skin metastases had developed and the possible influence of the different treatments administered. MATERIAL AND METHODS: The study was completed by determining the histopathological pattern, the immunohistochemical profile, the type of immunoglobulin and immunoglobulin light chain produced by the neoplastic plasma cells that infiltrated the skin, as well as the cytogenetic characteristics of these cells. RESULTS: From a clinical standpoint, the skin lesions consisted of multiple nodules or plaques with erythematous or violaceous coloration, and variable location. Histopathologically, two patterns were seen: nodular and diffuse interstitial. The plasma cells showed cytological atypia, and in one case they displayed a spindle shape, giving the lesion a sarcomatoid appearance. Immunohistochemically, these neoplastic plasma cells were strongly positive for CD79a, CD138, and EMA, while the immunoexpression of CD38 and CD43 was less intense and constant. In 39 of the 40 patients, the type of monoclonal immunoglobulin produced by the neoplastic plasma cells in the skin lesions was determined: 18 patients had IgA myeloma (5 IgAkappa and 13 IgAlambda), 19 patients had IgG myeloma (15 IgGkappa and 4 IgGlambda) and 2 patients had IgDlambda myeloma. The 22 cases in which immunoglobulin heavy chain gene rearrangement studies by PCR were carried out for the JH gene showed monoclonal rearrangement, while viral studies to try to identify genetic material of the HHV 8 virus and the Epstein-Barr virus gave negative results in all cases. These 22 cases studied using FISH showed the deletion of the rb-1 retinoblastoma gene in the neoplastic plasma cells that infiltrated the dermis. Despite aggressive chemotherapy, all of the patients died a few months after the skin lesions developed. CONCLUSIONS: In our series, a perfect correlation was seen between the findings of serum electrophoresis and the immunohistochemistry of the skin lesions with regard to the type of immunoglobulin and the immunoglobulin light chain restriction produced by the neoplastic plasma cells. Patients with multiple myeloma have a very short survival period once specific skin lesions appear, regardless of the therapy administered. The deletion of the rb 1 gene may be a prognosis marker to identify those patients with especially aggressive forms of multiple myeloma. PMID- 16476271 TI - [Prurigo pigmentosa]. AB - Prurigo pigmentosa is an infrequent inflammatory dermatosis of unknown etiology, characterized by recurrent episodes of pruritic erythematous papules which develop into intense reticulated pigmentation. It has been referenced most frequently in young women in Japan. Only thirty cases have been described outside of Japan. We describe a 32-year-old Spanish female who developed a pruritic dermatosis with the clinical characteristics and histopathological findings of prurigo pigmentosa. We review the epidemiological, clinical and histopathological characteristics of this peculiar skin disease. PMID- 16476272 TI - [Nodular primary cutaneous amyloidosis associated with Sjogrens syndrome: presentation of a case]. AB - Nodular primary cutaneous amyloidosis is the least frequent clinical form of the cutaneous amyloidoses. It may be associated with myeloproliferative disorders, as well as with systemic amyloidosis. Its association with other entities, such as Sjogren's syndrome, has recently been described. We present the case of a female patient with Sjogren's syndrome who developed nodular primary cutaneous amyloidosis. PMID- 16476273 TI - [Acneiform eruption from epidermal growth factor receptor inhibitors]. AB - One of the fundamental aims of oncological research is the search for molecules with greater efficacy against tumors and less toxicity than the usual chemotherapeutic agents. Epidermal growth factor receptor inhibitors are a new group of drugs which, because of their more specific effect against neoplastic cells, seem to meet these characteristics. Skin eruptions are one of the most frequent adverse effects associated with their use, secondary to the drug's direct inhibitory effect on homeostasis of the epidermis and of the pilosebaceous follicle. Several cases of cutaneous toxicity in patients treated with epidermal growth factor receptor inhibitors have recently been published. We present three cases of acneiform eruptions attributable to different drugs in this family (cetuximab, gefitinib and erlotinib). PMID- 16476274 TI - [Segmental anhidrosis associated with Adie's pupil: an incomplete case of Ross syndrome]. AB - We present the case of a 43-year-old woman who presented with a localized area of anhidrosis on the upper left trunk and ipsilateral upper limb. It was associated with Adie's tonic pupil and an area of compensatory hyperhidrosis on the contralateral side. After 9 years of follow up, the alterations have remained stable. These alterations of the autonomous nervous system correspond to an incompletely expressed Ross syndrome. PMID- 16476275 TI - [Acquired paraneoplastic hypertrichosis lanuginosa]. AB - Acquired hypertrichosis lanuginosa, an infrequent, paraneoplastic process, consists of the rapid development of lanugo-type hair, primarily on the face. We describe a 50-year-old woman with a 4-month case of hypertrichosis on the face and upper body, and weight loss over the last 6 months, in whom an inoperable squamous cell carcinoma of the cervix was discovered. Hypertrichosis lanuginosa is an indicator of a poor prognosis. It is associated with tumors in any location, but most often with lung and colorectal tumors. This is the first case associated with squamous cell carcinoma of the cervix. PMID- 16476276 TI - [Eccrine squamous syringometaplasia from chemotherapy extravasation]. AB - Eccrine squamous syringometaplasia is a rare entity that occurs in patients who have received chemotherapy treatment. It also occurs in association with chronic ulcers, skin tumors, and after the ingestion of drugs, exposure to toxic agents and inflammatory processes. We present the case of a 47-year-old male diagnosed with microcytic lung carcinoma and superior vena cava syndrome. After receiving the third cycle of chemotherapy, and after accidental extravasation, he presented with an erythematous plaque. The histopathological study revealed eccrine squamous syringometaplasia together with interphase dermatitis and epidermal changes that may be responsible for the epidermal necrosis. PMID- 16476278 TI - [Perianal Bowen's disease treated with imiquimod]. AB - Bowen's disease is a special form of squamous cell carcinoma in situ that usually develops in photoexposed areas of skin. The treatment of choice is surgery. Less frequently, it may appear in other locations such as the nails, glans penis, intertriginous areas and the perianal region, where conventional treatment may be complicated. We contribute a new case of perianal Bowen's disease, which responded to treatment with imiquimod with no evidence of clinical recurrence after 3 years of follow up. Imiquimod appears to be a therapeutic alternative for perianal Bowen's disease, which seems to have a particular tendency to recur in this location where surgery may also be complicated. PMID- 16476277 TI - [Nail disorders from epothilone]. AB - We present the cases of two patients diagnosed with non-microcytic lung carcinoma who were treated with epothilone, a new chemotherapeutic agent which disrupts microtubule dynamics. The first one developed nail symptoms consisting of painful nails with a hemorrhagic appearance, which developed into significant structural disorders and onycholysis of all of the fingernails. The patient required antibiotic treatment for the associated paronychia and surgical removal of some of the nails. In the second case, asymptomatic dyschromia of the toenails appeared, with no inflammatory component. These findings show the similarity of the adverse effects of epothilone with those of the taxanes. PMID- 16476279 TI - [Bullous lesions on the back of the hands]. PMID- 16476280 TI - [Generalized hyperpigmented, hyperkeratotic plaques]. PMID- 16476281 TI - [Multiple unilateral lichen striatus in an adult]. PMID- 16476282 TI - [Woolly hair nevus]. PMID- 16476284 TI - [Long-term follow up of angiofibromas treated with CO2 laser in 23 patients with tuberous sclerosis]. AB - INTRODUCTION: Tuberous sclerosis is an autosomal dominant disease in which hamartomas form in multiple organs. Cutaneous changes are one of the primary characteristics of this disease. These include angiofibromas (AF), a common form of presentation that causes significant cosmetic and medical problems. The CO2 laser has been used satisfactorily in treating these lesions, but there are few studies that evaluate its long-term results. The aim of our study is to assess the long-term response of the treatment of angiofibromas. METHODS: A retrospective study was carried out on 23 patients with angiofibromas treated with CO2 laser. The patients were treated between 1991 and 2000, inclusive, with continuous or superpulsed CO2 laser. We classified the angiofibromas by size, initial treatment results and patients' ages (< 20 years and 20 years or older). RESULTS: Ages ranged from 12 to 39 years, with a median age of 22.5 years. After treatment, patients were followed up for a period of six months to 10 years. In the long-term analysis, we found that 30.1 % maintained the initial result, and 60.9 % showed different degrees of recurrence, with a mean recurrence time of 3 years. When we analyzed the long-term results by the size of the angiofibromas, initial result and patients' ages, we found no statistically significant differences among the different groups. The survival analysis of the age groups, with Kaplan-Meier curves, showed that the youngest patients (< 20 years) had earlier recurrences than the older ones (logarithmic range 4.01 and p < 0.05). CONCLUSIONS: CO2 laser treatment achieves good short-term results. On the other hand, one of the biggest problems is recurrence over the long term; this is probably due to the fact that, because of their nature, these lesions cannot be eliminated permanently. This work coincides with earlier studies which found no factors that would make it possible to predict the recurrence of the lesions. However, we can conclude that recurrence takes place at a later date in older patients, and therefore they have better cosmetic results over the long term. PMID- 16476283 TI - [Herpes simplex prophylaxis in facial exfoliation treatments]. AB - In recent years, a spectacular increase has taken place in the demand for techniques to improve the appearance of the skin, both for cosmetic and reparative purposes. The techniques used may have complications that interfere with the healing process, and therefore may affect the results from an aesthetic standpoint. The most significant complication is herpes simplex infection. Furthermore, accurate, early diagnosis of herpes infection makes greater individual and community control possible, thanks to the current availability of effective antiviral agents. The scientific literature relating to herpes simplex prophylaxis was reviewed, and this information was pooled with the knowledge and experience of a multidisciplinary group of experts. After all of this data was analyzed, the consensus was reached that all patients should be treated with specific oral medications for herpes simplex as a preventive measure. PMID- 16476285 TI - [Bibliometric study of the journal Actas Dermo-Sifiliograficas (1984-2003). I. Production analysis]. AB - OBJECTIVE: Analyze the scientific articles published in Actas Dermo Sifiliograficas (AD) between 1984 and 2003. The main bibliometric parameters analyzed were: number of articles, their length and type, primary subject areas, authorship of the documents, main contributing centers, departmental areas and geographic distribution of the articles. MATERIAL AND METHODS: All of the scientific articles published in the regular issues of the journal Actas Dermo Sifiliograficas between 1984 and 2003 (two decades), corresponding to volumes 75 to 94, were reviewed manually. RESULTS: A total of 2,604 articles were published in the journal AD during the period studied, with a yearly average of 130 articles. Clinical cases or notes were the main type of document (66.2 %), followed by original articles (23.5 %) and review articles (4.7 %). The length of the documents varied from 1 to 33 pages, with an average of 4.55 pages. The subjects that were most discussed in the documents from this period were dermatopathology (15.9 %) and therapeutics (15.4 %). A total of 11,667 authorships were found, with 2,907 different authors. The average number of authors per article for the period was 4.48. 56.7 % of the authors were occasional contributors (having signed a single document), while 3.5 % were top contributors, having signed 20 or more works over this period. Seven Autonomous Communities accounted for 85.5 % of the articles. The Community of Madrid was by far the largest contributor (917 documents, 35.2 %), followed by Andalusia (422, 16.2 %), Catalonia (240, 9.2 %) and Castile-Leon (221, 8.5 %). The 2,604 documents in the study were from 293 different centers, which accounted for 2,977 co-authorships. Most of these co-authorships were from large hospitals or hospital complexes. The most noteworthy of these were the Madrid hospitals 12 de Octubre (281 co-authorships) and Clinico San Carlos (153), and Hospital Universitario San Cecilio in Granada (149). Dermatology was the specialty of most of the authors (90.8 % of those authors listed first). The main specialty collaborating with dermatology was pathology, taking part in 28.5 % of the articles. CONCLUSIONS: An analysis of the productivity of the journal AD over two decades gives us an idea of scientific activity in dermatology in Spain, because of the journal's quantitative and qualitative significance in the context of Spanish dermatology. PMID- 16476286 TI - [Xanthomas in a patient with Langerhans cell histiocytosis and liver cirrhosis]. AB - Skin involvement in acute forms of Langerhans cell histiocytosis (LCH) is in the form of erythematous papules, although rare forms of xanthomatous lesions have been described. We present the case of a boy with acute disseminated LCH who, at the age of 16 months, began to experience outbreaks of seborrheic dermatitis-like skin lesions and progressive hepatic dysfunction. The symptoms were complicated by partial central diabetes insipidus and specific pulmonary infiltration by Langerhans cells, which led to fibrosis. During the course of the disease, the patient developed liver cirrhosis, alterations in the lipid profile and disseminated xanthomatous skin lesions, concomitant with the lesions specific to the LCH. Despite successive cycles of chemotherapy, the outcome was the death of the patient after five years, due to his liver disease. Xanthomatous lesions in LCH are typical of the late stages of chronic progressive forms, such as Hand Schuller-Christian disease. When they appear in acute disseminated forms, there is some controversy over whether they correspond to a progression of the disease towards more chronic forms, or whether they are associated independent lesions, such as in this case. PMID- 16476287 TI - [Long-standing scrofuloderma]. AB - We present the case of a 26-year-old Ecuadorian woman who presented with multiple hypertrophic scars along the lower left limb, secondary to a cutaneous process that she had had for 11 years, previously diagnosed and treated in her country as cutaneous leishmaniasis. It had been clinically worsening for 9 months with suppuration through the scars in the groin and thigh, associated with local pain and systemic symptoms. As we suspected cutaneous tuberculosis, we performed a number of complementary examinations to confirm the diagnosis and rule out bone involvement. Finally, a lymph node culture confirmed that it was scrofuloderma. PMID- 16476288 TI - [Endogenous ochronosis: a case description]. AB - Endogenous ochronosis or alkaptonuria is an autosomal recessive disease caused by a deficiency of the enzyme homogentisic acid oxidase. Affected individuals excrete high levels of homogentisic acid in the urine, which darkens when it is alkalinized or oxidized. Deposits of blackish-brown pigment also occur in connective tissue; this causes, usually starting around the age of 40, the typical external manifestations of this disease, along with disorders in other organs. We present a clinical case of endogenous ochronosis, a very infrequent disease in our milieu. We will discuss the most noteworthy features of the case. PMID- 16476289 TI - [Reflex sympathetic dystrophy: description of a case with skin lesions]. AB - Reflex sympathetic dystrophy or algodystrophy is a poorly defined syndrome in which the patient develops pain disproportionate to the cause. It is included among the complex regional pain syndromes. The symptoms are triggered by some type of trauma, at times trivial, and consist of burning pain, edema, changes in skin color, alterations in vascularization, temperature changes, hyperhidrosis and skin disorders, which primarily consist of atrophic changes. Other less frequent cutaneous manifestations have been described in patients with this syndrome. These include papules, blisters, inflammatory lesions and reticulated hyperpigmentation. We discuss the case of a patient with reflex sympathetic dystrophy who presented with superficial ulcers on the affected limb, which mimicked dermatitis artefacta. PMID- 16476290 TI - [Epidermodysplasia verruciformis]. AB - We present the case of a 50-year-old Latin American woman who consulted her physician because of recent pruritic lesions on her arms and thighs. During the examination, we observed multiple flat papules on the limbs, as well as hypopigmented macules on the trunk which, according to the patient, began to appear during childhood. A histological study was performed on both types of lesions, and showed some enlarged keratinocytes with light blue cytoplasm in the upper layers of the epidermis. More than 20 types of HPV associated with EV (HPV EV) have been described. Although it was previously thought that these were specific to EV, new molecular biology techniques have made it possible to isolate HPV-EV sequences in skin diseases, both benign and malignant, with epidermal hyperproliferation in the immunocompetent population. PMID- 16476291 TI - [Contact dermatitis from Agave americana]. AB - Numerous plant species and their derivatives can cause skin reactions through a variety of mechanisms: irritative contact dermatitis, allergic contact dermatitis, contact urticaria and photodermatitis. We present a case of irritative contact dermatitis after exposure to the sap of Agave americana. The skin symptoms in this case have only been described on rare occasions; although this condition usually presents with a papulovesicular rash, in this patient it appeared as purpuric lesions in the contact area. PMID- 16476292 TI - [Celecoxib-induced aquagenic keratoderma]. AB - Aquagenic keratoderma is an infrequent condition characterized by translucent, smooth-surfaced papules and plaques and prominent eccrine ducts. It is limited to the palms and appears or becomes more pronounced after exposure to water. Histopathologically, hyperkeratosis and dilation of the eccrine ducts are seen. This condition has been described in adolescents and young women. We present the case of a 31-year-old woman with rheumatoid arthritis who had begun treatment with celecoxib one month before the onset of her cutaneous symptoms. Similar changes have been described in association with cystic fibrosis, and a case induced by rofecoxib has also been reported. Higher levels of sodium in the skin associated with celecoxib could increase the keratin's ability to take in water, and this may cause the clinical symptoms. PMID- 16476293 TI - [Atypical aquagenic keratoderma]. AB - Aquagenic keratoderma is an infrequent condition characterized by the appearance of lesions on the palms of the hands a few minutes after contact with water; these lesions vanish a short time after they dry. Published cases were primarily in adolescent females. We present the case of a male patient with lesions that clinically corresponded to aquagenic keratoderma, except that they were distributed on the backs of both hands and on the anterior face of the wrist. These characteristics had not been described in the literature prior to this case. Previously described cases were reviewed and several differential diagnoses were established, such as rofecoxib-induced palmar lesions, or those appearing in cystic fibrosis. PMID- 16476294 TI - [Recurring macules and papules on the lower limbs of a 42-year-old female patient]. PMID- 16476295 TI - [Fast-growing pearly tumor on the right forehead]. PMID- 16476296 TI - [Imiquimod for the treatment of skin metastases of melanoma]. PMID- 16476297 TI - [Longitudinal melanonychia in pregnancy]. PMID- 16476298 TI - [Buerger's disease (thromboangiitis obliterans)]. AB - Thromboangiitis obliterans (TAO) is a segmental, inflammatory, vasoocclusive disease that predominantly affects the small and medium-sized arteries and veins of the extremities. It most often occurs in young male smokers, especially those from Mediterranean and Asian countries. It is considered an autoimmune process related to the use of tobacco products. Clinically, it is characterized by the presence of painful, ischemic ulcers of the digits. Histopathological studies usually show an occlusive intraluminal thrombus with a predominantly acute inflammatory infiltrate. To make the diagnosis, it is important to exclude other causes of ischemia of the extremities, and different authors have proposed criteria to establish this diagnosis. Treatment is only effective if it is accompanied by abstention from tobacco. There are also different pharmacological and surgical strategies for its management. PMID- 16476299 TI - [Bibliometric study of the journal Actas Dermo-Sifiliograficas(1984-2003) II. Analysis of bibliographical references]. AB - OBJECTIVE: To quantify the number of references for each of the articles published in the regular issues of the journal Actas Dermo-Sifiliograficas (AD) between 1984 and 2003, and to calculate the average number of references for each of these years. To study the consumption of information by the authors of documents published in AD through the analysis of all of the bibliographical references listed in the articles published in 1984, 1993 and 2003. MATERIAL AND METHODS: The number of references for each of the scientific articles published in the regular issues of the journal Actas Dermo-Sifiliograficas between 1984 and 2003 was reviewed manually. For the years 1984, 1993 and 2003, the type of document, language, country of origin and age of the references were analyzed. RESULTS: The 2,604 articles published in the journal AD between 1984 and 2003 provided 56,144 references. The average number of references per article for the entire period was 21.56 +/- 21.2. The type of document with the most references was the review (67.06 +/- 59.9), followed by original works (24.76 +/- 19) and clinical cases (17.95 +/- 9.7). The primary type of document for references in the three years studied was the review article, which went from 84.8 % in 1984 to 95.4 % in 2003. References to books, on the other hand, dropped from 10.6 % in the first year to 3.3 % in the last one (p < 0.001). The United States was the country of origin of a large part of the references reflected in the three years studied (55.4 %). Following at a great distance were the United Kingdom (15.7 %) and Spain (9 %). English was the language in which most of the references were written in the three years studied, increasing from 72.9 % in 1984 to 87.5 % in 2003. Spanish was the second most used language in the references (9.1 %); contrary to what was expected, its use decreased over the three years. The main journals referenced by the authors published in AD were the American publications Archives of Dermatology (12.3 %) and Journal of the American Academy of Dermatology (10.3 %). References to AD itself accounted for 4 % of the total; it was the fourth most referenced journal after the British Journal of Dermatology (7.4 %). CONCLUSIONS: Spanish dermatologists primarily use English-language publications for their research. They mostly obtain current information from periodicals, with a significant percentage of the information being found in a small group of journals, which are the ones with the greatest international impact for our field of specialization. PMID- 16476300 TI - [Bibliometric study of Actas Dermo-sifiliograficas (1984-2003) III. Analysis of bibliographic impact factors]. AB - OBJECTIVE: To quantify the impact factor of the journal Actas Dermo Sifiliograficas (AD) from 1986 to 1990 and from 1999 to 2003 and to identify the journal's citation pattern in those years. MATERIAL AND METHODS: Citations obtained by AD in the periods from 1985-1990 and 1998-2003 for articles published from 1984 to 1989 and from 1997 to 2002 were collected using Science Citation Index Expanded (SCI). RESULTS: The number of times AD was cited doubled during the second period, increasing from 38 (period from 1985-1990) to 76 (period from 1998-2003). Considering the number of citations, AD's impact factor increased from 0.016 in 1986 to 0.040 in 2003. In both periods, citations corresponding to AD articles were included in a wide range of source journals, mainly dermatological publications abroad. The most referred journals in the second period were the Dutch publication Journal of the European Academy of Dermatology and Venereology (13 citations) and the Journal of the American Academy of Dermatology (12 citations). Unlike the period from 1985 to 1990 when no Spanish journal cited AD, four Spanish publications mentioned AD in the second period: Revista clinica espanola (6 citations), Archivos de bronconeumologia (4 citations), Medicina clinica (3 citations) and the journal Enfermedades infecciosas y microbiologia clinica (1 citation). Citations mainly corresponded to articles published by Spanish authors (63.2 % in the 1985-1990 period and 81.6 % in the period from 1998 to 2003). Self-citation increased from 10.5 % (first period) to 31.6 % (second period). CONCLUSION: The impact factor of AD is low and not comparable to other publications included in the Dermatology and Venereal Diseases field from SCI. Our results confirm the low citation rate of AD by source journals in this repertory. However, the increase of this rate in recent years seems to indicate a higher Spanish presence in SCI due to an increasing number of publications corresponding to Spanish authors in international journals and the inclusion of some Spanish journals in this database. PMID- 16476301 TI - [Neutrophilic eccrine hidradenitis secondary to thioguanine in a neutropenic patient]. AB - Neutrophilic eccrine hidradenitis (NEH) is an infrequent, self-limited inflammatory dermatosis characterized by a neutrophilic infiltrate around the eccrine glands. Clinically, it presents with different types of lesions. NEH occurs most frequently in patients who have undergone chemotherapy for hematologic neoplasms. We present a case of NEH in a 70-year-old neutropenic male who received thioguanine for acute myeloid leukemia. The erythematous plaques disappeared in 3-4 weeks. The histological findings were compatible with NEH. Skin cultures ruled out infectious causes. PMID- 16476302 TI - [De Sanctis-Cacchione syndrome]. AB - We present a male patient with photosensitivity since the earliest months of his life, and pigmented macules in exposed areas, some showing clinical atypia, which increased in number over time. Molecular biology studies detected an alteration in DNA repair ability, so xeroderma pigmentosum was diagnosed. Shortly after birth, low weight, microcephaly and psychomotor retardation had been observed, but the cause was not established. The patient progressively showed neurological disorders that included perceptive deafness, hyporeflexia and areflexia, as well as choreoathetotic movements. Therefore, we felt that the patient's symptoms fit De Sanctis-Cacchione syndrome. PMID- 16476303 TI - [Stevens-Johnson syndrome/toxic epidermal necrolysis treated with intravenous immunoglobulins]. AB - Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are part of the same disease spectrum, but are differentiated by the degree of skin detachment. In TEN, more than 30 % of the body's surface area is affected; thus, it is a serious process, whose frequency is estimated at 1.2-6 cases per million population/year. We describe the case of a 75-year-old male who suffered from SJS which evolved into TEN, probably because of the ingestion of ginkgo biloba extract. He was treated with intravenous immunoglobulins (Ig IV) at a dose of 0.5 g/kg/day for five consecutive days, with favorable progress and no significant side effects. It is evident that isolated cases do not justify the systematic use of this treatment, but they may help build up experience. PMID- 16476304 TI - [Cutaneous marginal zone B-cell lymphoma treated with rituximab]. AB - Marginal zone B-cell lymphoma (MZL) is probably the most frequent of the primary cutaneous B-cell lymphomas, which are entities with indolent behavior. Clinically, it appears in middle-aged patients as papules, nodules or erythematous plaques, solitary or multiple, on the trunk and proximal part of the limbs. The prognosis is excellent despite frequent cutaneous recurrences. We present the case of a 40-year-old male who, after having several recurrences of MZL over a ten-year period, was treated with rituximab for multiple skin lesions. The patient showed full remission after four weeks of treatment, and developed cytokine-release syndrome after the first infusion of the drug. PMID- 16476305 TI - [Granuloma from liquid silicone]. AB - In recent years, the demand for cosmetic interventions to augment soft tissue by injecting different substances has increased, due to their apparent innocuity. However, these procedures are not free from adverse reactions, such as the formation of foreign body granulomas, a phenomenon described in literature with most of the materials used. We report the case of a female patient with inflammatory lesions of the face, whose diagnosis was made after the histopathological study, which revealed a granuloma caused by liquid silicone. PMID- 16476306 TI - [Angiosarcoma in an irradiated breast: a case description]. AB - Post-radiotherapy cutaneous angiosarcomas have been described in different locations, including the breast. We present a case of cutaneous angiosarcoma of the breast, diagnosed 6 years after a carcinoma of the breast had been treated with radiation. The patient was a 44-year-old female with a history of ductal carcinoma (CA) of the right breast treated with tumorectomy, axillary lymphadenectomy, chemotherapy (FEC) and radiotherapy, who 6 years later presented with a violaceous, indurated plaque with a satellite nodule on the same breast. The histology of the skin biopsies showed angiomatous proliferation throughout the entire dermis, with no cellular atypia. Considering the patient's symptoms, history and the changes observed via mammography, it was decided to completely excise the lesion followed by a simple mastectomy, with the diagnosis of angiosarcoma being confirmed. The patient was later treated with paclitaxel, and the disease was apparently controlled. Despite this fact, two years and one month later, the angiosarcoma recurred on the internal area of the mastectomy scar. The patient was treated with paclitaxel, and surgical excision of the lesion area was once again performed. This complication usually appears 5-10 years after treatment with radiotherapy, so angiosarcoma should be ruled out if any angiomatous lesions later appear on skin that had been irradiated. PMID- 16476307 TI - [Primary cutaneous diffuse large B-cell lymphoma of the leg according to the new WHO-EORTC classification. Two cases]. AB - Primary cutaneous lymphomas are a heterogeneous group of lymphoproliferative disorders characterized by skin involvement with no evidence of systemic disease at the time of diagnosis. Their clinical behavior is generally indolent, and only occasionally is the development of extracutaneous disease observed. Since the 1980s, primary cutaneous B-cell lymphomas have been considered a specific group of lymphomas, differentiated from both T-cell lymphomas and from secondary cutaneous B-cell lymphomas. Both the EORTC and the WHO have proposed alternative classifications for these entities, with significant discrepancies that were finally resolved through the development of a new classification (WHO-EORTC classification for cutaneous lymphomas), which standardizes criteria that had previously been different. We present two new cases of primary cutaneous diffuse large B-cell lymphoma of the leg according to the new classification. PMID- 16476308 TI - [Basal cell carcinoma overlying a dermatofibroma]. AB - The epidermis over a dermatofibroma may show changes that range from simple hyperplasia to the proliferation of basaloid cells, which can become morphologically indistinguishable from basal cell carcinoma. The existence of a true basal cell carcinoma overlying a dermatofibroma is infrequent. These basaloid proliferations have usually been considered to be the result of the inductive effect of the fibrohistiocytic proliferation of the dermatofibroma on the epithelial cells of the hair follicle; therefore, it would be a reactive phenomenon and not truly neoplastic. We describe a case of dermatofibroma that presented with a basaloid proliferation identical in appearance to a basal cell carcinoma on the overlying epidermis. PMID- 16476309 TI - [Allergic contact eczema from epoxy resin]. AB - Epoxy resins are plastics that are widely used as electrical insulation, in coatings, and as adhesives and paints. They have strong sensitizing power and are one of the main causes of allergic contact eczema, both in the workplace and elsewhere. We present the case of a worker at a plastics/chemical plant, who handled aeronautical components in the process of manufacturing fuselage parts. He consulted his physician because of eczematous lesions on his fingers, hands and forearms which had developed over a two-year period and were clearly related to his work. The standard battery of skin tests was performed, along with the plastics and adhesives series and tests using the products from his workplace. Positivity was shown to epoxy resins (standard battery) and to the products from his workplace, which included different fiberglass and carbon fiber sheets impregnated with epoxy resins and epoxy adhesives. PMID- 16476310 TI - [Scleromyxedema: ultrastructural study]. AB - Scleromyxedema is a type of mucinosis that presents with some well defined clinical and histopathological characteristics. We describe the findings observed with transmission electron microscopy in a case that we recently studied. The patient was a 56-year-old female with a localized eruption of lichenoid papules on the face, upper trunk and limbs. It was accompanied by IgG lambda gammopathy. Optical microscopy showed the findings typical of scleromyxedema. Electron microscopy revealed the existence of a large number of fibroblasts with high activity levels in the synthesis and release of collagen fibers and a mucoid substance. PMID- 16476311 TI - [Tumor involving the back]. PMID- 16476312 TI - [Nodule on the pinna]. PMID- 16476313 TI - [The sentinel node biopsy from an immunological perspective: unjustified reservations]. PMID- 16476314 TI - [Strict anatomical coexistence of vitiligo and psoriasis]. PMID- 16476315 TI - [Consensus document on phototherapy: PUVA therapy and narrow-band UVB therapy]. AB - It is essential to develop a consensus document on phototherapy in order to adapt this procedure to the specific characteristics, needs and reality of our milieu. Using a review of existing literature on the subject and the experience of its own members as a reference, the Spanish Photobiology Group (GEF) of the Spanish Academy of Dermatology and Venereology (AEDV) has developed some therapeutic guidelines for the most widely used modes of phototherapy: PUVA therapy and narrow-band UVB (NBUVB) therapy. These guidelines deal with generalities about the equipment, calibration and regulation in phototherapy booths, and the concept and indications for these forms of treatment are reviewed. Recommendations are also proposed regarding patient selection, therapeutic procedures, associated pharmacological agents of interest and the prevention and management of adverse effects. The consensus document is designed as a flexible and practical instrument intended for use in daily clinical practice, aimed at optimizing the possibilities of phototherapy while reducing risks for patients and therapists. PMID- 16476316 TI - [Validation of the Spanish version of the Psoriasis Disability Index questionnaire in assessing the quality of life of patients with moderate-severe psoriasis]. AB - INTRODUCTION: To assess the measurement properties of the Spanish version of the Psoriasis Disability Index (PDI) quality of life questionnaire for patients with moderate or severe psoriasis in ordinary clinical practice. PATIENTS AND METHODS: A study was carried out with 294 patients with moderate or severe psoriasis who had received therapeutic treatment. Sociodemographic and clinical variables were collected and the PDI and EuroQoL-5D questionnaires were administered at the baseline visit, and at 3 and 6 months later. The feasibility (percentage of patients responding to the questionnaire), validity of the construct, internal reliability and sensitivity to change were analyzed. RESULTS: Feasibility: 98.6 % of the patients answered over 80 % of the questions on the questionnaire. Validity of the construct: the PDI scores were correlated with the scores obtained in the Psoriasis Area Disability Index (PASI) (r = 0.33) and the Visual Analogue Scale (VAS) of the EQ-5D (r = -0.41) (p < 0.01). Longitudinal validity: the PDI questionnaire score after 6 months of treatment was correlated with the changes in the PASI index and the VAS (r = 0.39 y -0.51). Reliability: the internal consistency of the dimensions was high (Cronbach's alpha = 0.89). Sensitivity to changes: the size of the effect corresponding to the changes experienced by the patients who noticed an improvement in the severity of their psoriasis during the period between visits was 0.95. CONCLUSION: The Spanish version of the PDI has proven to be reliable, valid and sensitive to changes for use with patients with psoriasis in the Spanish population, although the test retest reliability should be analyzed for clinically stable patients. PMID- 16476317 TI - [Treatment of cutaneous T-cell lymphomas with bexarotene]. AB - INTRODUCTION: The choice of treatment in cutaneous T-cell lymphomas (CTCLs) depends on the clinical stage of the disease and the patient's general condition. To date, there is no curative treatment for this disease, and the objective is to control the symptoms and prevent the disease from progressing. Bexarotene is an X receptor-specific retinoid with anti-tumor activity. Its use as treatment for CTCLs refractory to at least one prior systemic therapy has been approved by the FDA. PATIENTS AND METHODS: We carried out a descriptive study of 9 patients treated with bexarotene in the Lymphoma Unit of our department. We analyzed the clinical characteristics of the patients and the efficacy of the treatment, and we collected data on the side effects that appeared. RESULTS: The overall response to the treatment was 44.4% (4/9). 2 patients had full remission and 2 had partial remission. Tolerance to the treatment was good, and the most frequent side effects were hypertriglyceridemia, hypercholesterolemia and central hypothyroidism. CONCLUSIONS: Even though this is a series of only 9 patients, the results that we obtained are similar to ones previously described. Bexarotene is an effective therapeutic option in this heterogeneous group of diseases. PMID- 16476318 TI - [Adult-onset colloid milium. Presentation of two cases]. AB - Colloid milium is a degenerative process that is characterized clinically by the development of translucent, yellow, 1-2 mm papules located in photoexposed areas. Histologically, deposits of a colloid substance are seen in the papillary dermis. We present two cases of this infrequent pathology in two male patients, who had been subjected to intense sun exposure because of their work. We initiated treatment with photoprotective creams and topical tretinoin, with little clinical improvement. PMID- 16476319 TI - [Kindler syndrome: presentation of a case]. AB - Kindler syndrome is a very rare disease caused by mutations resulting in defects in the extracellular matrix-actin link. It usually presents with acral blistering from birth in trauma-prone areas, pronounced photosensitivity that improves with age and the development of poikiloderma and cutaneous atrophy. Mucosal involvement and degeneration have been described with relative frequency. PMID- 16476320 TI - [Pemphigus vulgaris beginning as the Koebner phenomenon]. AB - Pemphigus vulgaris is an autoimmune disease characterized by acantholysis and intraepidermal bullae formation. The isomorphic Koebner phenomenon is a response to different stimuli, and it has been described in many skin disorders. An association between the two has rarely been described. We present a case of pemphigus vulgaris that appeared over surgical scars. Pemphigus vulgaris over scars is very rare. In our case, the reactivation of the pemphigus antigens in the surgical scar probably caused a generalized autoantibody response, with the formation of bullae on the patient's trunk. PMID- 16476321 TI - [Hyperkeratotic vascular malformations. Presentation of three cases]. AB - Hyperkeratotic vascular malformations (verrucous hemangiomas) are infrequent vascular lesions present from birth. Initially, they are reminiscent of port wine stains or childhood hemangioma, but over time they gradually take on their typical warty and hyperkeratotic appearance. These changes are probably due to trauma, scratching and bleeding episodes. Because the lesion is deep and goes beyond the clinical lesion, treatment is difficult and involves many recurrences. We present three cases of this vascular malformation, in two males aged 45 and 62 and a 30-year-old female. All three had the lesion from birth, and presented with frequent bleeding episodes. PMID- 16476322 TI - [Neonatal lupus erythematosus: 4 cases and clinical review]. AB - Neonatal lupus erythematosus (NLE) is an infrequent disease in newborns caused by the transplacental passage of maternal Anti-Ro/SSA, Anti-La/SSB and/or Anti-U1 RNP antibodies. The most common manifestations are cutaneous and cardiac. We carried out a retrospective study of cases of NLE diagnosed in the last 10 years at the Hospital Universitario Insular in Gran Canaria. Complete data was obtained for 4 patients. Three cases had circulating Anti-Ro antibodies in the mother and in the newborns, while in the fourth case they were Anti-RNP. Two mothers were diagnosed with systemic lupus, one with mixed connective tissue disease and the other with leucocytoclastic vasculitis. The skin lesions consisted of urticaria like and desquamative lesions. One patient presented with ulceration. The histological study of the urticaria-like lesions showed a non-specific perivascular infiltrate; the desquamative lesions were consistent with subacute lupus erythematosus. PMID- 16476323 TI - [Focal epithelial hyperplasia]. AB - Focal epithelial hyperplasia (FEH) is a benign proliferation of the oral mucosa with well defined clinical and histological characteristics. It has been associated with infection of the oral mucosa by types 13 and 32 of the human papillomavirus (HPV), and to a lesser extent, with other types. Its clinical course is variable, although it usually persists for months or years; cases with spontaneous resolution have been described, as have others with prolonged persistence. We present the case of an Ecuadorian boy whose visit was motivated by lesions in the oral mucosa consistent with a diagnosis of FEH, which were confirmed in the histological study, and in which HPV type 13 DNA was identified. PMID- 16476324 TI - [Lentigo maligna treated with 5% imiquimod cream]. AB - Lentigo maligna (LM) is considered to be an in-situ stage of lentigo maligna melanoma. Clinically, it presents as a pigmented macule, irregular in shape and tone. 30% to 35% of untreated lentigo malignas can progress into lentigo maligna melanoma. The treatment of choice for this pathology is surgical excision with margins of 0.5 cm. of clinically normal skin around the lesion, or Mohs microsurgery. Imiquimod is a topical immunomodulator that stimulates both acquired and innate immunity. We present the case of a patient with LM, treated with 5% imiquimod cream, with an excellent therapeutic response. PMID- 16476325 TI - [Verrucous erosive plaques in the inguinal folds]. PMID- 16476326 TI - [Labial swelling which had developed over two months]. PMID- 16476327 TI - [Cutaneous leishmaniasis]. AB - Leishmaniases are diseases caused by infection by protozoa of the genus Leishmania. Cutaneous leishmaniasis caused by Leishmania infantum is frequent in Spain, especially in certain geographic areas. Diagnosis of cutaneous leishmaniasis is difficult because of the varied symptoms and because making cultures of this parasite is complicated. There are also different therapeutic, medical and surgical options, none of which is fully satisfactory. We review the most significant agents of cutaneous leishmaniasis in Spain. PMID- 16476328 TI - [Solar urticaria. Study of 20 cases]. AB - INTRODUCTION: Solar urticaria is an infrequent disorder, but is probably underdiagnosed. It is characterized by the sudden appearance of weals in areas that are not usually photoexposed after exposure to the sun or to artificial sources of visible or ultraviolet light. Few cases have been published in literature, so the information available about this disorder and its natural evolution is limited. MATERIAL AND METHODS: We have compiled data from 20 patients diagnosed with solar urticaria in our department in the last 12 years (1990-2002) in order to try to obtain information about the characteristics of this condition. RESULTS: As the most relevant characteristics of our series, we can mention the fact that 60 % of the patients were women, and the average duration of the condition before consulting a dermatologist was 3 years. In 55 % of the patients, areas that are usually photoexposed, such as the face and hands, were less severely affected (due to acclimatization or hardening). The spectra responsible for the SU were visible light, UVA and UVB, in that order. With regard to treatment, we obtained a partial response with antihistamines and photoprotectors, and good results using progressive desensitization with UVA/sunlight. CONCLUSION: Solar urticaria is probably an underdiagnosed condition in our milieu. There are few series in literature that provide any information about the most significant characteristics of this disorder. We have compiled the most relevant data from our patients and we have compared it with the other published series in an attempt to learn more about this photodermatosis. PMID- 16476329 TI - [Inhibition of the motility of melanoma cells using interference RNA against CD9]. AB - INTRODUCTION: Although 65 % of the deaths caused by skin cancer are due to melanomas, the possible causes that may explain the aggressiveness of this type of tumor are still unknown. Different approaches have been used to try to find an effective treatment, but they have been unsuccessful. Interference RNA (iRNA) is an essential technique for this research. With this technique, we can selectively "knock down" or silence protein expression. The tetraspanins CD9 and CD151 are molecules involved in cell motility, including melanoma cells. MATERIAL AND METHODS: A375 melanoma cell line, anti-CD9 and anti-CD151 monoclonal antibodies, and iRNA against CD9 and CD151. Immunofluorescence techniques, flow cytometry, cell transfection, cell selection with magnetic beads and evaluation of cell migration in a wound-healing model. RESULTS: The A375 cells express CD9 and CD151. By using iRNA against CD9 and CD151, we managed to inhibit the expression of these proteins. The cells transfected with iRNA against CD9 showed significant inhibition of their motility. DISCUSSION: We managed to silence the expression of CD9 and CD151 using iRNA techniques in the A375 melanoma cell line. The reduction in CD9 caused the inhibition of cell motility, while the interference with CD151 expression had a more moderate effect. This data indicates that the knockdown of tetraspanins may be a future target for the treatment of melanoma. PMID- 16476330 TI - [Focal acral hyperkeratosis associated with pitted keratolysis]. AB - Focal acral hyperkeratosis is characterized by the same clinical appearance as acrokeratoelastoidosis, but without abnormalities in the elastic fibers. We present the case of a woman with a 10-year case of dermatosis localized on the palms, soles and dorsum of the metacarpophalangeal joints, consisting of multiple polygonal papules and associated hyperhydrosis, clinically compatible with acrokeratoelastoidosis. Her father had a history of the disease. In addition, the patient presented with palmoplantar pitted keratolysis. The histopathological study ruled out elastorrhexis, and the pitted keratolysis was corroborated by the clinical appearance and the presence of coccoid elements in the stratum corneum, evident with a PAS stain. In our opinion, the focal acral hyperkeratosis is not a separate entity from the acrokeratoelastoidosis. PMID- 16476331 TI - [Childhood eosinophilic pustulosis]. AB - Childhood eosinophilic pustulosis is a rare disease that is characterized by recurrent outbreaks of pruritic pustules and follicular papules. The lesions are sterile and contain masses of eosinophils related to the scalp hair follicle. Because of the good prognosis for these symptoms, conservative treatment with topical corticosteroids is recommended. We present two cases of this disease, describing its clinical course and development over two years. PMID- 16476332 TI - [Umbilical cutaneous endometriosis associated with a large uterine myoma]. AB - We present a 44-year-old female patient who was being studied by the Gynecology Department because of the presence of large abdominal masses which proved to be leiomyomas on histological analysis. The Dermatology Department was consulted because of an asymptomatic umbilical tumor which had been developing for 5 years. Upon histological examination, we observed glands with angular lumens that showed decapitation secretion distributed throughout the dermis, surrounded by a cellular stroma with extravasated erythrocytes. PMID- 16476333 TI - [Scleredema of Buschke associated with diabetes mellitus. Study of four cases]. AB - Scleredema of Buschke is characterized by a thickening of the dermis with mucin deposits among the collagen fibers, which manifests as a hardening of the skin, predominantly on the upper trunk. It has been associated with long-standing, poorly controlled diabetes mellitus, monoclonal gammopathies and streptococcus infections. We studied patients with a clinical diagnosis of scleredema confirmed by means of a histopathological study, and associated with diabetes mellitus. We reviewed associated diseases, clinical and histopathological characteristics, evolution and response to treatment. Scleredema is characterized by hardening of the skin, affecting the upper trunk. A large proportion of cases are associated with long-standing, poorly controlled diabetes mellitus, with little response to treatment. PMID- 16476334 TI - [Pigmented apocrine hidradenoma]. AB - Apocrine hidradenoma is a benign adnexal neoplasm. It has no specific site predilection, and usually affects middle-aged people. The same as other tumors of the sweat glands, there is a pigmented variety. We present the case of a 92-year old male who consulted his physician for a slow-growing asymptomatic lesion in the right groin which had been developing for 2 years. After the histopathological study, the diagnosis was established as pigmented apocrine hidradenoma. PMID- 16476335 TI - [Family Buschke-Ollendorff syndrome]. AB - Buschke-Ollendorff syndrome is an autosomal dominant disorder whose clinical course includes elastic nevi and osteopoikilosis. Histologically, most cases present with a normal amount of collagen in the skin lesions and an increase in elastic fibers, although abortive forms with skin involvement have been described, with a decrease in elastic fibers and an absence of bone alterations. We describe the case of a 41-year-old woman with Buschke-Ollendorff syndrome criteria, and that of her 2-year-old daughter who might present an abortive form of Buschke-Ollendorff. PMID- 16476336 TI - [Infectious bacteremic ecthyma in a patient with HIV infection]. AB - Ecthyma is a deep ulcerative pyoderma usually caused by group A beta-hemolytic streptococcus (Streptococcus pyogenes). It usually presents in areas of damaged skin and in immunocompromised patients, and very infrequently is accompanied by general symptoms and bacteremia. We describe the case of a 41-year-old female patient with HIV infection who presented with fever and ulcerated lesions on the right ankle and left foot which had been developing for three weeks. Streptococcus pyogenes and Escherichia coli were simultaneously isolated from a culture of the skin biopsy sample and from blood cultures. Escherichia coli is of dubious pathogenic significance in this case. PMID- 16476337 TI - [Occupational allergic contact eczema in a dental assistant]. AB - Occupational contact eczemas are very frequent in dental assistants. We present the case of a 46-year-old woman with pruritic, flaky, erythematous-edematous lesions, with indistinct borders, on the backs and sides of the fingers, which had been developing for several months. The skin tests performed with the GEIDC (Spanish Contact Dermatitis Research Group) standard battery, the Chemotechnique dental screening series and her own products showed positivity to nickel sulfate, cobalt chloride, potassium dichromate, gold sodium thiosulfate, the liquid used in endodontics known as "eugenol," fragrance mix and colophony. Eugenol is a perfumed product that is used in cements and sealants for endodontics and periodontology. There are 5 articles regarding cases of occupational allergic contact eczema from this product in dentists or nurses and assistants who work in dental clinics. PMID- 16476338 TI - [Diffuse darkening of the skin]. PMID- 16476339 TI - [5-year-old yellowish-brownish tumor of the back]. PMID- 16476340 TI - [Regarding the variability of medical practice in skin cancers]. PMID- 16476341 TI - [Systemic amyloidoses]. AB - Amyloidoses are a heterogeneous group of diseases characterized by extracellular fibrillar protein deposits in the organs and tissues. These proteins are not biochemically related to each other, but share certain common characteristics, including apple green birefringence with polarized light after staining with Congo red, and beta-pleated sheet configuration through x-ray diffraction. Amyloid deposits may occur in many organs (systemic amyloidoses) or may affect a single tissue (localized or organ-specific amyloidoses). There are different classifications, but in this review the amyloidoses are organized by clinical symptoms, which are determined by the amyloid protein involved. Special attention is given to cutaneous and mucous membrane manifestations, which are often the first sign of the disease and are useful for early diagnosis, thus avoiding more aggressive procedures. The involvement of other organs is analyzed, as are the diagnosis, prognosis and treatment of systemic amyloidoses. PMID- 16476342 TI - [Prospective study of the levels of serum cytokines in patients with melanoma: prognostic value]. AB - INTRODUCTION: The production of cytokines plays a primordial role in the immune systems fight against tumors. Therefore, we proposed to investigate whether the serum levels of different types of cytokines in melanoma patients were associated with the evolution of their disease. MATERIAL AND METHODS: We analyzed the variation in the serum levels of cytokines representative of the Th1 (INFg and IL 15) and Th2 (IL-6 and IL-10) phenotypes in 33 patients with primary cutaneous melanoma. Peripheral blood samples were obtained every six months until we had a total of 4 samples per patient. RESULTS: After 30 months, 29 patients (87.9 %) had survived with no signs of recurrence. Basal IL-10 serum levels were higher in the group of patients who expired than in the survivors. Among the patients who expired, an increase in IL-6 serum levels was observed in the last sample. No relationship could be proven between INFg and IL-15 levels and melanoma progression. CONCLUSION: Determining the levels of type Th2 cytokines (IL-6 and IL-10) in the serum of melanoma patients could be useful in the clinical follow up of these patients and serve as a predictive factor for the progression of the disease, with the prognosis being worse for patients with high IL-10 and IL-6 levels. On the other hand, determining the serum levels of type Th1 cytokines (IL 15 and INFg) does not seem to be as useful in predicting the prognosis. PMID- 16476343 TI - [Diagnostic cost-effectiveness of the skin biopsy in inflammatory diseases of the skin]. AB - INTRODUCTION: Inflammatory skin diseases are a very heterogeneous and extensive group of entities whose clinical and pathological diagnosis is difficult. In cases where the clinical diagnosis is doubtful, the histopathological study of one or more lesions can be of great help. The aim of our work is to compare the effectiveness of the histopathological diagnosis in inflammatory skin lesions according to the department performing the biopsy. MATERIAL AND METHODS: A retrospective study was carried out on the reports for pathological study requests and the histopathological reports from the year 2003 from the Pathology Department of Hospital Santa Maria del Rosell. In the reports for the pathological study requests, the following data was assessed: department performing the biopsy, whether or not the type of lesion was stated on the request form, location, evolution and clinical diagnosis. The histopathological diagnoses were reviewed by one of the authors and classified into two major groups: a) specific diagnoses and b) non-specific diagnoses. Five departments took part in performing biopsies on inflammatory skin lesions: Dermatology, General Surgery, Primary Care, Internal Medicine and Gynecology. To better compare the data, we have divided the departments into two groups: 1. Dermatology Department and 2. Non-dermatology departments. We performed a statistical analysis (proportion comparison) of the specific diagnoses between the two groups and among the specific diagnoses of the non-dermatology departments. RESULTS: The total number of inflammatory skin lesions studied was 97. The Dermatology Department performed 48 biopsies, and the non-dermatology departments performed 49. There was less clinical data in the reports sent by the non-dermatological departments than in those from the Dermatology Department. The pathologist made a specific diagnosis in 77 % of the biopsies performed by the Dermatology Department, compared to 41 % of the biopsies sent by the non-dermatology departments (p < 0.001). There are no statistically significant differences among the specific diagnoses made by the departments that make up the non-dermatology group. CONCLUSION: The histopathological diagnoses made in the biopsies sent from the Dermatology Department are more specific (77 %) than those made in the biopsies performed by the non-dermatology departments (41 %). Thus, biopsies on inflammatory skin lesions should be performed by the Dermatology Department so that diagnostic effectiveness is maximized, and in order to prevent delays and inappropriate treatments that might be harmful to the patient. PMID- 16476345 TI - [Follicular mycosis fungoides, comedo-like and cystic]. AB - Follicular mycosis fungoides is an infrequent variant of mycosis fungoides. It has classically been defined by the presence of an atypical lymphoid infiltrate around and in the follicular epithelium with little or no epidermotropism, and no follicular mucin deposits. The fact that there are cases with epidermal involvement and/or follicular mucinosis means that some uniform diagnostic criteria are necessary. We describe two cases of follicular mycosis fungoides with follicular mucinosis and with varying degrees of associated epidermotropism. PMID- 16476344 TI - [Panniculitic T-cell lymphoma]. AB - Panniculitic T-cell lymphoma is a rare, aggressive variant of cutaneous T-cell lymphoma, with fewer than 100 cases described. The main problem is its diagnosis, as both the clinical and the histological features may simulate benign panniculitis. We present the case of a 34-year-old male patient, who had presented with an indurated plaque, sclerodermiform in appearance, on the front of the right thigh for 4 months, later accompanied by fever and constitutional symptoms. The initial diagnosis was cellulitis, but no clinical improvement was seen despite systemic antibiotic therapy. After two skin biopsies, the patient was diagnosed with panniculitic cutaneous T-cell lymphoma. The patient was treated with 8 cycles of CHOP chemotherapy, with resolution of the symptoms. PMID- 16476346 TI - [Merkel cell carcinoma. Study of five cases]. AB - Merkel cell carcinoma (MCC) is a rare malignant skin tumor of neuroendocrine origin with a poor prognosis and rapid progression. It is usually an erythematous nodule on the face, and is associated with other skin neoplasms. Its histology shows a large mass of small cells containing oval nuclei with powdery chromatin in the dermis. These cells are positive for neurospecific enolase, chromogranin, synaptophysin and cytokeratins AE1/AE3. We present five patients (3 males and 2 females, aged 58 to 89 years) seen at our hospital in the last three years for MCC. Their treatment and evolution were tracked, and many of the aspects indicated were seen. MCC must be kept in mind in the differential diagnosis, as early detection along with aggressive treatment significantly improve the patients survival rate. Close follow up is necessary because of the high recurrence rates. The development of a response protocol in order to better manage this disease would be desirable. PMID- 16476347 TI - [Topical treatment of melanoma skin metastases with imiquimod]. AB - The treatment of skin metastases of melanoma can be difficult in many cases because of the patients age, as well as the number, size and location of the lesions. We present the case of an 82-year-old male with melanoma skin metastases on the scalp, which responded satisfactorily to treatment with 5 % imiquimod cream. Imiquimod is a topical immunomodulator with antiviral and antineoplastic action. This case, along with others that have recently been published, supports the usefulness of this treatment in selected cases of melanoma skin metastases, at least for palliative purposes. PMID- 16476348 TI - [Toxic epidermal necrolysis caused by lamotrigine]. AB - Toxic epidermal necrolysis (TEN) is a serious entity consisting of a generalized eruption with the formation of flaccid blisters. It affects at least 30 % of the cutaneous surface and is usually related to the prior ingestion of medications. We describe TEN symptoms related to the introduction of lamotrigine, an anti epileptic drug prescribed in this case for the prevention of migrainous episodes. PMID- 16476349 TI - [Herpetiform pemphigus associated with esophageal carcinoma]. AB - We present the case of a 63-year-old woman with herpetiform pemphigus and disseminated esophageal carcinoma which began to manifest themselves simultaneously. Some cases of herpetiform pemphigus associated with neoplasms have previously been reported, but these have primarily been bronchopulmonary. The association of any type of pemphigus with esophageal cancer is very infrequent. We should be alert to the infrequent varieties of pemphigus and atypical bullous diseases, because they sometimes do not meet the diagnostic criteria of paraneoplastic pemphigus, but are the first manifestation of an underlying neoplasm. PMID- 16476350 TI - [Generalized eczema secondary to combined treatment with peginterferon alfa-2a and ribavirin in a patient with chronic hepatitis from the hepatitis C virus]. AB - Adverse skin effects from interferon (IFN) plus ribavirin combined therapy are relatively frequent, but they are usually local and related to the IFN injection site. However, distant or generalized eczematous reactions secondary to this treatment are rare. The introduction of pegylated interferons may increase the frequency of these skin lesions. PMID- 16476351 TI - [Yellowish plaques on the trunk and in the axillary folds]. PMID- 16476352 TI - [Nodules and violaceous plaques on the scrotum and thigh]. PMID- 16476353 TI - [Eruptive seborrheic keratoses triggered by eczema]. PMID- 16476354 TI - [Treatment of chronic ulcers]. AB - Chronic ulcers are a challenge in dermatological therapy. It is essential to establish their etiology in order to treat them, but on many occasions local therapy is of great interest. Treatment of chronic ulcers is currently based on so-called moist wound healing, and it takes two aspects into consideration: the underlying pathology and local treatment. Local treatment is always necessary and includes: cleaning, debridement, the control of any infection, and the application of different topical agents, both medication and dressings. Recently, new therapeutic strategies are being established, some of which are still being assessed, and which include: skin replacement using biological skin substitutes, growth factors, laser, hyperbaric oxygen, electrical stimulation and negative pressure dressings. In this work, we review the therapeutic advances in this pathology, without neglecting the validity of classic treatments. PMID- 16476355 TI - [Subjective perception of pain in local dermatological surgery]. AB - INTRODUCTION: Pain is difficult to measure, as it has a significant subjective component. This symptom, and in particular its psycho-emotional component (fear prior to the operation and a feeling of release afterwards), has not been studied extensively. We decided to quantitatively evaluate its perception in local surgery, in relation to different moments during the operation and using the application of a topical anesthetic cream as an external variable. PATIENTS AND METHODS: The perception of pain was documented with a verbal assessment scale before and during anesthesia infiltration and immediately at the end of the operation by means of three questions: 1) How much do you think it is going to hurt?, 2) How much does it hurt? and 3) How much did it hurt? In addition, the application of EMLA (topical anesthetic cream) was randomized. RESULTS: 219 patients were studied. The scores for question 2 were significantly lower than the ones for question 1, and the scores for question 3 were significantly lower than the ones for questions 1 and 2. The patients treated with EMLA only gave lower scores than the controls in questions 2 and 3. DISCUSSION: Patients think that the surgery will hurt more than it really does, and they also immediately erase the memory of the recent pain. The use of EMLA does not modify the patient's idea of how much the operation will hurt, but the patient does perceive less pain than those who do not use it. It is important to reassure patients about their operations so that they do not become overly anxious about the procedures, thus minimizing their perception of pain. The use of a topical local anesthetic in the area of the surgery could help decrease the sensation of pain. PMID- 16476356 TI - [Atypical fibroxanthoma. Clinical/pathological study of 10 cases]. AB - INTRODUCTION: Atypical fibroxanthoma (AFX) is a rare tumor of unknown histogenesis, considered by most authorities as a superficial form of malignant fibrous histiocytoma (MFH). The aim of this work is to report the clinicopathological features of 10 cases of AFX. MATERIAL AND METHODS: Data were retrospectively collected of the computerized medical history. Clinical (age, onset-diagnosis time, location, accompanying pathology, outcome), histological (architectural pattern, cell type, ulceration, vascular or perineural invasion, subcutis involvement, pleomorphism, mitosis, inflammatory infiltrate) and immunohistochemical variable were analyzed. CASES REPORT: Clinical and epidemiological features coincide with those previously reported: onset late in life, short time onset-diagnosis, involvement of skin with notable sun damage and a good outcome. Pathologically all the cases showed a spindle-cell prevalence arranged in a vaguely storiform pattern, along with both, multinucleated and eosinophilic cells. DISCUSSION: The diagnosis of AXF is always of exclusion. Other spindle-cell tumors such as squamous cell carcinoma, malignant melanoma, leyomiosarcoma or dermatofibrosarcoma protuberans must be ruled out by immunohistochemical techniques. In spite of its rarity, the recognition of AFX is important in order to avoid inappropriately aggressive treatment. PMID- 16476357 TI - [Vitiligo. Treatment of 12 cases with topical tacrolimus]. AB - INTRODUCTION: Vitiligo is a frequent dyschromia, characterized by achromic macules that reflect the absence of melanocytes. The cause of this selective destruction seems to be due to an autoimmune phenomenon. Tacrolimus is an immunomodulator produced by Streptomyces tsukubaensis, whose topical use has been approved for atopic dermatitis. It has been tested in other dermatoses where immunological phenomena are involved. MATERIAL AND METHODS: During the period from September 1, 2003 to April 30, 2004, an open study was carried out on 12 cases of vitiligo treated with topical tacrolimus 0.1 % twice a day. The degree of repigmentation was analyzed using digital photography at the initial visit, and at three and six months. The response in each case was taken into consideration, as well as the response by treated area. Possible adverse effects during the treatment period were also noted. RESULTS: 50 % of the patients treated showed repigmentation with good (50 %-75 %) or excellent (> 75 %) improvement after 6 months. All of the patients with facial involvement achieved repigmentation of over 50 % in this location. Repigmentation in all cases took place homogeneously and centripetally, rather than in a perifollicular pattern. Repigmentation began before three months of treatment had elapsed in 10 patients, and after three months in the remaining two. We did not find any evidence of adverse effects except pruritus in the eyelid area in two patients during the first week of treatment. CONCLUSION: We believe that tacrolimus 0.1 % applied topically for a minimum of six months may be a valid alternative in the treatment of vitiligo in the facial area, especially the eyelids, where other therapeutic modes are not recommended because of the possible side effects. PMID- 16476358 TI - [Lipoid proteinosis]. AB - Lipoid proteinosis is an infrequent disease characterized by the deposition of a PAS-positive diastase-resistant hyaline material in the skin and respiratory tract, although it can also be deposited in internal organs, in a generally asymptomatic manner. The earliest clinical manifestation is hoarseness. Clinical cutaneous manifestations come later, in the form of hyperkeratotic lesions located on the trunk, elbows, axillae, groins, backs of hands, palms and soles. A lesion typical of the disease is moniliform blepharosis, which consists of beaded papules along the eyelid margins. Also characteristic is the presence of comma shaped intracranial calcifications in the temporal lobes. The course of the disease is progressive, with a normal life expectancy. It affects men and women equally, with worldwide distribution. The diagnosis is based on the clinical symptoms and the histology. At this time, there is no effective treatment for the disease. We present a case of lipoid proteinosis in a 23-year-old woman, with typical clinical and histological characteristics. PMID- 16476359 TI - [Follicular mucinosis secondary to captopril-induced photoallergy]. AB - Captopril is an angiotensin-converting enzyme inhibitor which has been widely used as an antihypertensive agent and vasodilator. Although adverse skin effects occur in 10 % of patients prescribed this drug, there is little published data about photosensitivity. On the other hand, follicular mucinosis has been described in association with numerous processes, mainly lymphomas, lupus erythematosus and, less frequently, with photoinduced eruptions and drug reactions. We present the case of a female patient treated with captopril who developed an eruption in photoexposed areas. Histologically, the patient showed changes consistent with follicular mucinosis, with a positive captopril photopatch test and with resolution of the lesions after the antihypertensive agent was withdrawn. PMID- 16476360 TI - [Multiple adult xanthogranuloma]. AB - Juvenile xanthogranuloma is an infrequent, benign, normolipemic, non-Langerhans cell histiocytosis, which primarily affects young children. Clinically, it is characterized by the appearance of one or several brownish-yellow papulonodular lesions on the upper body, especially on the head and neck. Xanthogranulomas are less frequent in adults, and generally present as solitary lesions. The diagnosis of multiple xanthogranulomas in adults is exceptional. We describe the case of a female patient diagnosed with this rare disease. PMID- 16476361 TI - [Necrolytic migratory erythema associated with glucagonoma]. AB - Glucagonoma is a rare pancreatic tumor that is usually associated with a syndrome that includes diabetes, anemia, weight loss and skin lesions in the form of necrolytic migratory erythema. We present the case of a patient with malignant glucagonoma treated with surgery and octreotide, which manifested with skin lesions. The discussion will review the physiopathology, other causes of necrolytic erythema, diagnosis and differential diagnosis and treatment. PMID- 16476362 TI - [Cholesterol crystal embolism]. AB - Cholesterol crystal embolism (CCE) is an infrequent entity that primarily appears in males over the age of 60 with generalized arteriosclerosis after angiographic procedures, vascular surgery or, more rarely, with oral anticoagulant treatment with heparin or with fibrinolytics. We present the case of a patient with several risk factors for CCE, who presented with the pathognomonic triad of leg and foot pain, livedo reticularis and palpable pedal pulses. The diagnosis was based on the fact that cholesterol crystals were seen in the arterioles in the skin biopsy. Due to the frequency with which the skin manifestations appear and the importance of early diagnosis and treatment, an awareness of these crystals is fundamental in diagnosing these processes. PMID- 16476363 TI - [Lupus miliaris disseminatus faciei]. AB - Lupus miliaris faciei is a chronic inflammatory dermatosis that most often affects young adults. For years, it has successively been considered a form of tuberculosis, sarcoidosis and rosacea. Recently, some authors have proposed that it should be considered a distinct entity. It is an eruption of small, brownish erythematous papules, primarily located on the face, especially in the periocular area. It is self-limited and generally leaves residual punctate scars. We present the case of a 25-year-old male affected by Lupus miliaris disseminatus faciei and discuss its main characteristics, as well as the different problems in its etiopathogenic classification. PMID- 16476364 TI - [Cutaneous bronchogenic cyst]. AB - Bronchogenic cysts rarely affect the cutaneous surface, so they are difficult to diagnose clinically and are often confused with other types of cysts. We present the case of a young adult in whom a cystic lesion showing characteristics of a bronchogenic cyst was excised from the upper thorax area. We emphasize the fact that, in nearly all cases, the diagnosis of this pathology is made through a histopathological study, and we make special reference to the differential diagnoses. PMID- 16476365 TI - [Congenital botryoid rhabdomyosarcoma of the vulva]. AB - Rhabdomyosarcoma is the most frequent malignant soft tissue tumor in pediatric patients; however, the vulvar location and congenital appearance are exceptional. We present the case of a newborn girl with botryoid rhabdomyosarcoma of the vulva, treated with chemotherapy, conservative surgery and autologous transplant. Botryoid rhabdomyosarcoma is a variation of embryonal rhabdomyosarcoma that typically grows in mucosa-lined hollow organs, from where it can spread to the body surface. The treatment of botryoid rhabdomyosarcoma in the genito-urinary area is based on polychemotherapy, and it can be complemented with radiotherapy and conservative surgery if necessary, thus resulting in an excellent prognosis and few long-term functional sequelae. PMID- 16476366 TI - [Erythematous scaly facial plaques]. PMID- 16476367 TI - [Rownish-erythematous atrophic plaques, developed over 40 years]. PMID- 16476368 TI - [Fulminant meningococcemia]. PMID- 16476369 TI - [Multiple eruptive dermatofribromas in a patient with Down's syndrome]. PMID- 16476370 TI - [Biomolecular advances in hereditary epidermal disorders]. AB - In recent years, the genes responsible for many hereditary skin diseases have been discovered. These genes encode different proteins that participate in the terminal differentiation of the epidermis, so their alteration or absence causes a keratinization disorder and/or an increase in skin fragility. Thanks to genetic analyses, we have been able to understand the physiopathology of numerous genodermatoses and we have become closer to diagnosing many others. In the not too-distant future, biomolecular techniques may foreseeably help us prevent and treat these processes, which include skin diseases as serious as epidermolysis bullosa or epidermolytic hyperkeratosis. In this article, we will study the most recent biomolecular findings referring to keratinization and epidermal disorders, mentioning the altered genes and/ or the defective proteins that cause them. PMID- 16476371 TI - [Euromelanoma Day. Results of the 2000, 2001 and 2002 campaigns in Spain]. AB - BACKGROUND: In the year 2000, the European Academy of Dermatology and Venereology initiated a campaign, called "Euromelanoma Day," for the early detection of melanoma. OBJECTIVE: To discuss the results of "Euromelanoma Day" in Spain for the campaigns carried out in 2000, 2001 and 2002. MATERIAL AND METHODS: Participating members of the Spanish Academy of Dermatology examined everyone who requested an appointment, free of charge, in order to detect suspicious lesions. Four months after the visit, telephone contact was made with all patients who had been diagnosed with suspected melanoma to ask them whether the lesions had been excised, and if so, the result of the histological study of the specimen. RESULTS: 33,750 calls to set up an appointment were received, and 12,487 patients were examined. The average participation by dermatologists was 399 academy members per year. A total of 164 lesions suspected of being melanoma were detected, above all in Andalusia and Catalonia, and the diagnosis was histologically confirmed in 31 cases. The clinicopathological correlation was 23.3 %, a higher figure than the one for the USA and similar to the one for Mediterranean countries. The average thickness of the tumors was 0.93 mm. CONCLUSIONS: Although these campaigns have the drawback of possible bias in the findings and may cause some degree of public alarm, the benefit to the population was unquestionable in terms of heightening awareness of a health problem. PMID- 16476372 TI - [Evaluation of a screening system for patients with pigmented lesions using store and-forward teleconsultation]. AB - INTRODUCTION: Pigmented lesion clinics (PLC's) were developed as a quick referral system for patients with pigmented lesions. However, the most appropriate method of selecting patients who need to be seen in these units is not clearly defined. Teledermatology is a tool whose usefulness as a patient selection system for PLC's needs to be evaluated. OBJECTIVE: To evaluate teleconsultation as a screening system for patients with pigmented lesions in terms of efficacy, accuracy and satisfaction. METHOD: Teleconsultations received at a PLC over a period of 12 weeks were evaluated. Teleconsultation patients reported changes in a pigmented lesion, a lesion that had recently appeared, multiple lesions, symptomatic lesions or concern about a nevus. We calculated the time intervals in sending the teleconsultation report and in patients being seen at the "physical" PLC consultation, the intraobserver, interobserver and pathologist kappa coefficients, as well as the degree of satisfaction of patients and Primary Care (PC) physicians. RESULTS: 219 teleconsultations were evaluated, 49.3 % of which were referred to the "physical" consultation. The most frequent reason for the teleconsultation was concern about a nevus (37.0 %). The teleconsultations received responses in an average time of 43.9 hours, and patients were seen at the "physical" consultation within 2 weeks. The intraobserver agreement was kappa = 0.93 (95 % CI 0.87-0.98); interobserver agreement, kappa = 0.91 (95 % CI 0.87 0.96); and the agreement between the teledermatologist and the pathologist, kappa = 0.79 (95 % CI 0.70-0.89). 86 % of the patients and 91 % of the Primary Care physicians said that they were "very satisfied" with the implementation of this new system. CONCLUSIONS: Teleconsultation is an accurate screening system for patients with pigmented lesions. With this methodology, waiting times for patients with malignant lesions or those suspected of malignancy can be shortened at the same time as the PLC's excess workload is decreased. However, more experience is needed to establish the true usefulness of this filtering system in the early diagnosis of melanoma. PMID- 16476373 TI - [Mastocytosis in adults. Description of nine clinicopathological cases]. AB - INTRODUCTION: Mastocytosis is a hyperplastic process characterized by the infiltration of different organs and tissues by mature mastocytes. It is more frequent in childhood, although cases also occur in adults. There are significant differences between the ways mastocytosis presents in adults and children, as well as in its development and prognosis. MATERIAL AND METHODS: In this work, we describe the clinicopathological findings for 9 adult patients with mastocytosis. The clinical, evolutional and genetic characteristics of mastocytosis in adults are also compared to those of childhood mastocytosis. RESULTS: In contrast with childhood mastocytosis, the skin lesions of adult mastocytosis are very monomorphous, and consist of macules and papules of less than 1 cm in diameter, brownish-red in color. There are few symptoms, and there is usually discrete pruritus. Darier's sign is very often negative. Furthermore, the skin lesions do not tend to spontaneously regress, which is what usually occurs in children. Finally, systemic involvement in adults is practically a constant, with mastocyte infiltration of the bone marrow in over 90 % of cases, and bone involvement in over 50 % of cases, while systemic involvement is rarer in children. In any case, even if there is infiltration of systemic organs in adult mastocytosis, there are usually no accompanying clinical symptoms. The mutation of the c-kit proto oncogene consists of the replacement of Asp with Val at codon 816, and this is nearly always found in adult mastocytosis. It is less frequent in childhood mastocytosis. CONCLUSIONS: All of these findings make it possible to classify adult mastocytosis as a separate clinicopathological entity from mastocytoses in children. PMID- 16476374 TI - [Cutaneous large B-cell lymphoma of the leg]. AB - Large B-cell lymphoma of the leg (LBCLL) accounts for 2 % of primary cutaneous lymphomas, and the EORTC considers it to be of intermediate prognosis. It usually presents in elderly patients, and manifests clinically as erythematous nodules or tumors, often unilateral, on the lower third of the legs. On rare occasions, it presents with extracutaneous dissemination. We present the case of a 77-year-old male who consulted his physician because of an asymptomatic indurated erythematous papule which had appeared one month earlier on his left leg and had quickly and progressively increased in size. Concurrently, other similar lesions had been appearing on the thigh and back of the same leg. Histological analysis detected a large cell lymphoid infiltrate positive for CD20 and bcl-2 throughout the dermis. The extension study was normal. The patient was treated with an intravenous infusion of rituximab for four consecutive weeks, with full remission of the lesions. PMID- 16476375 TI - [Mechanic's hands: a characteristic cutaneous sign of antisynthetase syndrome]. AB - "Mechanic's hands" are a characteristic cutaneous sign of idiopathic inflammatory myositis. We describe the case of a 61-year-old male patient who was diagnosed with idiopathic polymyositis and non-specific interstitial lung disease in 1999, and three years later developed scaly, fissured hyperkeratotic lesions on the lateral and palmar surfaces of the first three fingers of both hands, with little pruritus. The presence of the anti-Jo-1 antisynthetase antibody in the patient's serum, the finding of skin lesions characteristic of "mechanic's hands" and the patient's other systemic clinical manifestations made it possible to establish the diagnosis of "antisynthetase syndrome." PMID- 16476376 TI - [Two cases of leprosy in Valladolid]. AB - Leprosy is still a major health problem on a worldwide level, and although it does not reach endemic levels on our continent, it seems that there is an upward trend due to the number of imported cases diagnosed each year. We present two cases of lepromatous leprosy diagnosed in Valladolid within a five-month period. The last reported case of this disease in our community was over 20 years ago. PMID- 16476377 TI - [Subacute cutaneous systemic lupus erythematosus. Presentation of two clinical cases]. AB - We present two cases of systemic lupus erythematosus (SLE) which began with subacute clinical manifestations. Both cutaneous and systemic lupus are the result of interactions between susceptibility genes and environmental factors such as ultraviolet radiation, giving rise to an anomalous response with hyperreactivity of T and B lymphocytes. Over 50 % of subacute cutaneous lupus cases have or will have SLE, while only 16 to 61 % of SLE cases have acute cutaneous lupus lesions. PMID- 16476378 TI - [Acneiform eruption secondary to cetuximab]. AB - C225 (cetuximab) is a monoclonal antibody that targets the epidermal growth factor receptor (EGF-R). It is used for the treatment of solid malignant tumors in advanced stages. It works against tumors by inhibiting cell proliferation, angiogenesis and the formation of metastases, as well as by promoting cell apoptosis. We present the case of a 64-year-old male patient affected with a colon neoplasm with hepatic metastases, for which treatment with cetuximab was indicated. He came to our department because of a skin eruption with papules and pustules located on the face, neck, presternal area and upper back, but with no cysts or comedones. The biopsy was compatible with an acneiform eruption. The patient was treated with minocycline, 100 mg/day for 2 weeks, with the clinical symptoms responding favorably. When he was given further doses of cetuximab, he once again presented with new eruptions, but of lesser intensity. Because of the high frequency with which this adverse effect appears, it is recommended that cetuximab be included on the list of drugs causing acneiform eruptions. PMID- 16476379 TI - [Bullous pemphigoid in an infant after vaccination]. AB - Bullous pemphigoid (BP) is a disease that characteristically affects the elderly, although cases have been described in children. We present a case of BP in a two month-old infant with bullous lesions on the palms and soles, which appeared one week after receiving the first dose of the hepatitis B, polio, DTP and HiB vaccine. She was treated with sulphated water, dexamethasone and fusidic acid, and the lesions disappeared. One month later, she presented with a new, more generalized outbreak, three days after the second dose of the same vaccine. The skin eruption completely subsided 3 months after treatment with deflazacort was initiated (1 mg/kg/day). After five years of follow up, the patient has not presented with lesions again, despite having received the rest of the vaccines on the official schedule. PMID- 16476380 TI - [Pyoderma gangrenosum over a saphenectomy scar in a cANCA-positive patient with diverticulitis]. AB - Pyoderma gangrenosum is a chronic, painful, ulcerative skin disease of unknown etiology. In 50 % of the cases, it is associated with systemic disease, primarily with inflammatory bowel disease. It can develop over areas of trauma or surgical wounds, possibly because of local immunoreactant deposits. We describe the case of a cANCA-positive patient with colonic diverticulitis who developed pyoderma gangrenosum over a saphenectomy scar. PMID- 16476381 TI - [Occupational allergic contact dermatitis from color developers used in automatic developing]. AB - The color developers CD2, CD3 and CD4 and the black-and-white developer TSS are well known contact allergens that affect those who work in photographic processing. The frequency of occupational allergic contact dermatitis from color developers has decreased in recent years because of the fact that most photo developing takes place automatically, reducing exposure to different chemicals used in developing. We present a case of occupational allergic contact dermatitis in a non-atopic male who had worked for the last 5 years in a photography shop equipped with an automatic developing machine. The clinical morphology of the lesions was eczematous, and the patch tests showed contact sensitization to CD2, CD3, CD4 and TSS. The clinical symptoms cleared up when direct contact with the developing product containing CD3 and CD4 was avoided. PMID- 16476382 TI - [Fast-growing frontal erythematous plaque]. PMID- 16476383 TI - [Verroucous papules on the back of the hands]. PMID- 16476384 TI - [Management in daily practice]. PMID- 16476386 TI - [Multinucleate-cell angiohistiocytoma]. PMID- 16476387 TI - [Langerhans cell histiocytosis]. AB - Histiocytoses originate from the proliferation of mononuclear phagocytes in different tissues. These entities make up a heterogeneous group, and are mainly classified as Langerhans cell histiocytoses and non-Langerhans cell histiocytoses. Langerhans cell histiocytoses have as a common characteristic the proliferation of dendritic antigen-presenting cells with phenotypical and ultrastructural characteristics of Langerhans cells. Infiltration may be limited to one organ, or may be disseminated. The prognosis and the treatment especially depend on the age of the patient and the number and dysfunction of the organs involved. Its etiopathogenesis is unknown, although most researchers currently believe that an alteration in the regulation of the immunological system occurs in these patients. PMID- 16476389 TI - [Study of the significance of female dermatologists in the Spanish Academy of Dermatology and Venereology]. AB - BACKGROUND: Most Spanish dermatologists are members of the Spanish Academy of Dermatology and Venereology (AEDV). Their development and history can be considered to parallel each other. The aim of our study is to determine the role women have played in the AEDV throughout its history, both in number and as officers or directors. MATERIAL AND METHODS: To obtain our data, we consulted AEDV's files from the time it was founded until the year 2002. We also asked other medical associations, obtaining a response from three of them. RESULTS: No women joined the AEDV until 1964. After that date, the percentage rose slowly (1975, 5.5 %), until the last decade when it increased more sharply, reaching 41.34 % in 2001. There are only 11 women on the boards of directors, all in non presidential roles. The percentages of women in Pediatrics, Ophthalmology and Pathology are similar. CONCLUSIONS: The number of female dermatologists in the AEDV has increased gradually, although not steadily, reaching a maximum in the last decade. Women are rarely officers or directors, and there are no female presidents. The data shows that women have gradually become more prevalent in the medical profession, the same as in the rest of society as a whole. PMID- 16476388 TI - [Study on patient satisfaction with the treatment of mycosis on the extremities with terbinafine (SETTA)]. AB - INTRODUCTION: The SETTA study is an open-label, multicenter, prospective, observational study carried out in order to evaluate satisfaction with the treatment of mycosis on the extremities with terbinafine. METHODS: Patients clinically diagnosed with tinea pedis or tinea unguium after a single visit were recruited. Follow up was through a questionnaire that the patient was to send back. The two groups were statistically compared by means of three analyses: descriptive, effectiveness and safety. RESULTS: In the effectiveness analysis, significant differences were observed in the evolution of the tinea, the overall score and degree of adherence to the treatment, with these being more positive in the group with tinea pedis. In the safety analysis, adverse events, none of them serious, were reported in 24 patients. CONCLUSIONS: This study reflects a high degree of satisfaction and safety in the use of terbinafine to treat patients with mycosis on the extremities. PMID- 16476390 TI - [Clinically-amyopathic dermatomyositis: presentation of three cases]. AB - Dermatomyositis is a connective tissue disease which has several factors involved in its etiology. Recently, the existence of a sub-type called amyopathic dermatomyositis has been accepted, whose cutaneous manifestations are indistinguishable from the ones that appear in the classic form of the disease. However, it is different from the classic form in that clinical findings of muscular involvement are initially absent. The association of these two sets of symptoms with the possible presence of an underlying neoplastic process has been described on numerous occasions, and has been the subject of extensive debate. We present three patients with amyopathic dermatomyositis, two males aged 27 and 45 and a female aged 54, the latter associated with a malignant process, and we discuss and review the main differential features of the entity and its relationship with different types of neoplasms. PMID- 16476391 TI - [Amyopathic dermatomyositis associated with a recurrence of breast cancer]. AB - The term "amyopathic dermatomyositis", or dermatomyositis "sine myositis" is used to describe those patients who present with the skin manifestations typical of dermatomyositis, but with no evidence of inflammatory myopathy. Amyopathic dermatomyositis may be associated with an underlying neoplasm, the same as with classic dermatomyositis. We present the case of a 59-year-old female patient, with cutaneous findings typical of dermatomyositis, with no proximal muscle weakness and with normal serum muscle enzymes, which stayed in a normal range throughout the later follow-up period, although the electromyogram performed six months later showed alterations with a myopathic pattern. These skin symptoms raised the suspicion of an occult neoplasm, and a recurrence of the patient's breast cancer, apparently inactive for many years, was finally found. The association of amyopathic dermatomyositis with a recurrence of breast cancer is exceptional. PMID- 16476392 TI - [Idiopathic atrophoderma of Pasini and Pierini. Study of 4 cases]. AB - Idiopathic atrophoderma of Pasini and Pierini (IAPP) is an infrequent form of cutaneous atrophy, of unknown etiology. We describe the clinical and histological symptoms of four women with IAPP. Three of them had lesions on the back, while the fourth had them on the buttocks. The histological study of two patients revealed a variable degree of collagen alteration (sclerosis and hyalinization). In the other patients, the dermis was apparently normal. There is discussion about whether IAPP is a distinct entity or is related to morphea. Both sets of symptoms, although separate, could form part of the same spectrum within the fibrosing type of dermatoses. We found no positive serology for B. burgdorferi in our patients. PMID- 16476393 TI - [Normolipemic plane xanthomas and mycosis fungoides]. AB - Diffuse normolipemic plane xanthomas are characterized by the presence of yellowish plaques on the eyelids, neck, upper trunk, buttocks and flexures. Histology shows foamy histiocytes in the dermis. Approximately half of all cases are associated with hematological disorders. On rare occasions, they have been described in the context of cutaneous T-cell lymphomas. We present the case of a female patient with tumor-stage mycosis fungoides who developed normolipemic plane xanthomas coinciding with the appearance of new lymphoma lesions. We review English-language literature regarding the rare association of xanthomas and cutaneous T-cell lymphomas. PMID- 16476394 TI - [Superficial angiomyxoma]. AB - Superficial angiomyxomas are clinically variable, infrequent, benign skin lesions. They are usually located on the trunk, but can also appear on the lower limbs, head and neck. They are diagnosed histologically, and are characterized by the fact that they are poorly delimited, multinodular tumors, and by the presence of interstitial myxoid material with scanty cellularity. Treatment is through surgery, and local recurrence is possible. An associated Carney's complex must always be ruled out with superficial angiomyxoma. We present the case of a woman with a solitary, fast-growing superficial angiomyxoma. PMID- 16476395 TI - [Sweet's syndrome with bullous lesions]. AB - We present a case of Sweet's syndrome with atypical lesions, characterized by erythematous plaques, vesicles and bullous lesions. Skin lesions in patients with an underlying malignancy are more frequently atypical and with vesicular, bullous or even ulcerative characteristics, in addition to the typical plaques and nodules. However, the case presented is not associated with malignancy, despite the fact that these processes, particularly hematologic ones, should be suspected. PMID- 16476396 TI - [Localized acquired cutis laxa associated with trachyonychia]. AB - Acquired cutis laxa is a rare disease of unknown cause, which affects elastin metabolism. Clinically, it is characterized by redundant skin and hyperelasticity, while the histological study shows a reduction in or absence of elastic fibers in the dermis. We present a case of localized acquired cutis laxa associated with trachyonychia. PMID- 16476397 TI - [Lupus miliaris disseminatus faciei associated with epidermal cysts]. AB - Lupus miliaris disseminatus faciei (LMDF) is a chronic granulomatous disease, which tends to be located on the central part of the face. Its typical histological image is one of epithelioid granulomas with caseous necrosis. Its association with epidermal cysts has been described on few occasions in literature. We present a case of LMDF associated with epidermal cysts in a female Caucasian patient. This association suggests that the rupture of the epidermal cysts may be related to the appearance of LMDF. However, there are probably other factors that may also contribute to its pathogenesis. PMID- 16476398 TI - [Fast-growing tumor on the scalp of a 78-year-old woman]. PMID- 16476399 TI - [Painless nodules on the fingers of a 4-year-old boy]. PMID- 16476400 TI - Prevalence of infant Vitamin A deficiency and undernutrition in the Republic of Congo. AB - OBJECTIVES: A representative sample of 5722 pre-school children living in rural and urban areas of the Congo was examined between July and September 1999 for assessing Vitamin A deficiency. METHODS: Using a randomized two-level cluster sampling method, 190 clusters of 30 children aged from 6 months to 6 years were selected in order to assess the prevalence of active xerophthalmia (night blindness and/or Bitot spots). Concurrently, the children's height and weight were determined. A semi-quantitative seven-day dietary questionnaire was applied to the mothers of 5722 children to estimate the latter's consumption of Vitamin A rich foodstuffs. The prevalence of biochemical deficiency was assessed based on the serum retinol concentrations analyzed in dried blood spots from a sub-sample of 300 children living in the Pointe-Noire area. RESULTS: Among the 5722 children studied, 0.7% were found to suffer from night blindness and 7.7% had Bitot spots. The weekly intake of Vitamin A rich foods was estimated in 5722 children. Our data suggest that Vitamin A rich food consumption was lower in rural zones than in urban area according to the food frequency method threshold values. The serum retinol levels were lower than 10 microg/dl in 18% (95% confidence interval [C.I.]: 13.7, 22.3) [8.04+/-2.87 microg/dl] and less than 20 microg/dl in 49% (95% C.I.: 43.4, 54.6) [15.05+/-2.76 microg/dl] of the 300 studied children. We have established a significant relation between mean serum retinol levels and high rate of Vitamin A food intake (chi-square=59.64, 2 d.d.l., p<0.05) in the sample studied. The mean serum retinol concentrations did not differ significantly between the various Z-scores of weight for age (W/A) and height for age (H/A) patterns. But children with a weight for height (W/H) ratio below -2 standard deviation (S.D.) had significantly lower serum retinol values [9.33+/ 1.3 microg/dl] than those with a W/H ratio greater than or equal to -2S.D. [10.82+/-4.84 microg/dl]. CONCLUSION: These data suggest that Vitamin A deficiency is still a serious public health problem in rural areas of the Congo in which this study was carried out. PMID- 16476401 TI - Calculating molar absorptivities for quinones: application to the measurement of tyrosinase activity. AB - The molar absorptivities of the quinones produced from different o-diphenols, triphenols, and flavonoids were calculated by generating the respective quinones through oxidation with an excess of periodate. Oxidation of these substrates by this reagent was analogous to oxidation by tyrosinase with molecular oxygen, although the procedure showed several advantages over the enzymatic method in that oxidation took place almost immediately and quinone stability was favored because no substrate remained. The o-diphenols studied were pyrocatechol, 4 methylcatechol, 4-tert-butylcatechol, 3,4-dihydroxyphenylalanine, 3,4 dihydroxyphenylethylamine, 3,4-dihydroxyphenylacetic acid, 3,4 dihydroxyphenylpropionic acid, and caffeic acid; the triphenols studied were pyrogallol, 1,2,4-benzenetriol, 6-hydroxydopa, and 6-hydroxydopamine; and the flavonoids studied were (+)catechin, (-)epicatechin, and quercetin. In addition, the stability of the quinones generated by oxidation of the compounds by [periodate]0/[substrate]0 << 1 was studied. Taking the findings into account, tyrosinase could be measured by following o-quinone formation in rapid kinetic studies using the stopped-flow method. However, measuring o-quinone formation could not be useful for steady-state studies. Therefore, several methods for following tyrosinase activity are proposed, and a kinetic characterization of the enzyme's action on these substrates is made. PMID- 16476402 TI - Two-dimensional HPLC on-line analysis of phosphopeptides using titania and monolithic columns. AB - Conventional and comprehensive two-dimensional (2D) HPLC systems using the combination of titania and monolithic columns were established for the on-line analysis of phosphopeptides. Compared with immobilized metal affinity chromatography of a general method for the analysis of phosphopeptides, the use of titania columns in the analysis permits the specific isolation of phosphopeptides in a higher yield. Using the current 2D HPLC systems, phosphopeptides were specifically isolated from nonphosphorylated peptides by the first-dimension titania column, followed by the high-speed separation of the phosphopeptides by the second-dimension monolithic column. Proteolytic digests of beta-casein were analyzed within 30 min using the comprehensive 2D HPLC system; all phosphopeptides from beta-casein could be efficiently isolated and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The comprehensive 2D HPLC system coupled with mass spectrometry will be useful for high-throughput and on-line phosphoproteome analyses. PMID- 16476403 TI - Size exclusion chromatography-multiangle laser light scattering analysis of hyaluronan size distributions made by membrane-bound hyaluronan synthase. AB - Size exclusion chromatography-multiangle laser light scattering (SEC-MALLS) analyses of Escherichia coli membranes expressing Streptococcus equisimilis hyaluronan synthase (seHAS) demonstrated an inherent artifact (10-100 MDa) that coeluted with hyaluronan (HA) and skewed the apparent weight-average mass of HA to erroneously high values. Briefly heating samples to 65-75 degrees C eliminated this artifact and increased the yield of recovered HA due to the release of HA chains that were attached to membrane-bound HAS. Inclusion of alkaline phosphatase, which removed uridine 5'-diphosphate (UDP) produced during the reaction, improved the linearity of HA synthesis-even at high substrate use. Surprisingly, the addition of EDTA, to chelate Mg(2+) ions, did not completely stop the HAS reaction at 30 degrees C or at 4 degrees C. The best conditions for stopping the reaction without altering SEC-MALLS profiles of the product HA were to chill samples on ice in the presence of both EDTA and UDP. Even with excess substrate, the maximum size of product HA decreased as the enzyme concentration increased. Therefore, the maximum HA size made by HAS was determined by extrapolation to zero enzyme concentration. Using the above conditions, membrane bound seHAS synthesized a cohort of HA products that steadily increased in weight average molar mass, reaching a final maximal steady-state size of 4 to 6 MDa within 2-4 h. PMID- 16476405 TI - Producing and dispensing small quantities of 15N2 gas at atmospheric pressure. PMID- 16476404 TI - Reproducibility of gas chromatography-mass spectrometry measurements of 2H labeling of water: application for measuring body composition in mice. AB - Deuterium-labeled water (2H2O) has emerged as a novel isotope tracer. Following the administration of 2H2O, it is possible to study the dynamics of carbohydrate, protein, lipid, and DNA and to determine body composition. Those studies require reliable measurements of the 2H labeling of water. Although simple gas chromatography-mass spectrometry (GC-MS) methods have been developed for measuring the 2H enrichment of biological fluids, investigators have not reported on the intra- and/or interdaily variability of the measurements. We have experimentally examined the reproducibility of one GC-MS method for measuring the 2H labeling of water. Briefly, hydrogen (deuterium) atoms in water were exchanged with those bound to acetone, and the 2H labeling of acetone was then determined under electron impact ionization. We found that the coefficient of variation is generally less than 0.5% when water is labeled between 0 and 2.8 mole percentage excess 2H. We demonstrated that this highly reproducible result allows one to use 2H2O and the "acetone method" to measure physiological parameters such as body composition in mice. PMID- 16476406 TI - Fluorescently labeled collagen binding proteins allow specific visualization of collagen in tissues and live cell culture. AB - Visualization of the formation and orientation of collagen fibers in tissue engineering experiments is crucial for understanding the factors that determine the mechanical properties of tissues. In this study, collagen-specific fluorescent probes were developed using a new approach that takes advantage of the inherent specificity of collagen binding protein domains present in bacterial adhesion proteins (CNA35) and integrins (GST-alpha1I). Both collagen binding domains were obtained as fusion proteins from an Escherichia coli expression system and fluorescently labeled using either amine-reactive succinimide (CNA35) or cysteine-reactive maleimide (GST-alpha1I) dyes. Solid-phase binding assays showed that both protein-based probes are much more specific than dichlorotriazinyl aminofluorescein (DTAF), a fluorescent dye that is currently used to track collagen formation in tissue engineering experiments. The CNA35 probe showed a higher affinity for human collagen type I than did the GST-alpha1I probe (apparent K(d) values of 0.5 and 50 microM, respectively) and showed very little cross-reactivity with noncollagenous extracellular matrix proteins. The CNA35 probe was also superior to both GST-alpha1I and DTAF in visualizing the formation of collagen fibers around live human venous saphena cells. Immunohistological experiments on rat tissue showed colocalization of the CNA35 probe with collagen type I and type III antibodies. The fluorescent probes described here have important advantages over existing methods for visualization of collagen, in particular for monitoring the formation of collagen in live tissue cultures over prolonged time periods. PMID- 16476407 TI - Performance of the five-point grip pen in three screen-based tasks. AB - This study examined how user handbreadth, brace shape and brace size of a Five Point Grip Pen affects the performance of three screen tasks: writing, point-and clicking, and drawing. Nine braces with three shapes (ball, ball-and-brick, and natural) and three brace sizes (large, medium and small) were tested for performance time, number of errors and subjective preference by 32 subjects (16 males and 16 females). The subjects were selected to fit four groups of handbreadth (below 25%ile, 25%ile-50%ile, 50-75%ile and above 75%ile) of Taiwanese laborers. In general, the subjective ranking results were consistent with the results of performance measures in all three tasks. All measures were affected by brace shape and brace size but differently for each task. The study results can be summarized as follows: (1) The natural shape was the best brace for all tasks. (2) For the writing task, the small brace was best for all four handbreadth groups, the medium brace was adequate only for the handbreadth group of above 75%ile, and the large brace was worst for all groups. (3) For the drawing task, the large and medium braces were suitable for all handbreadth groups, while the small brace was not suitable for the handbreadth group of above 75%ile. (4) For point-and-clicking task, the large size was the best brace. PMID- 16476408 TI - A combined field and laboratory investigation for the effective application of ergonomics in situ. AB - Despite extensive research on musculoskeletal disorders associated with manual labour, the enormity of the problems experienced in industry remains. Recognizing the importance of applying the science of ergonomics, the focus of this paper was to highlight the substantial difference between conducting rigorous controlled research in the laboratory and the less controlled, but more realistic research within the working environment. Our proposal is not to use one or the other methodology, but rather to combine basic assessments made in situ with rigorous laboratory experimentation investigating human responses both pre- and post intervention, and finally to go back into the field to test the efficacy of the proposed ergonomics intervention. The combined 'field-lab-field' format presented in this paper is based on research conducted in an industrially developing country, and it is argued that this is the most likely means of assuring that the application of rigorous ergonomics theory will improve the poor working conditions so evident in developing regions. PMID- 16476409 TI - Amygdala volume and depressive symptoms in patients with borderline personality disorder. AB - BACKGROUND: Borderline personality disorder (BPD) is characterized by a high prevalence of comorbid psychiatric disorders, including major depression (MD). The aim of this study was to examine whether a co-occurrence of MD is associated with structural changes in the amygdala of BPD patients. METHODS: Twenty-five right-handed, female patients with BPD and 25 matched healthy control subjects were examined. Diagnoses of BPD and MD were made according to DSM IV. Depressive symptomatology was determined with the Hamilton Depression Scale (HAMD). Magnetic resonance imaging scans were performed with 1.5 T Magnetom Vision (Siemens, Erlangen, Germany). The software program "BRAINS" was applied for brain volumetry and segmentation. The amygdala was delineated as "region of interest." RESULTS: Comparison of amygdala volumes between the whole group of BPD patients and control subjects revealed no significant difference. Amygdala volumes in both hemispheres were significantly larger in BPD patients with MD compared with those without MD. There was a significant correlation in BPD patients between left amygdala volume and depressive symptoms as measured by HAMD. CONCLUSIONS: Correlation of amygdala volume with depression in BPD patients might indicate a causal relationship. Future studies should clarify whether amygdala enlargement is a risk factor for MD in BPD patients or a consequence of the affective disorder. PMID- 16476410 TI - RETRACTED: The Relationships Between White Matter Cerebral Metabolites and Clinical Manifestations in Geriatric Patients with Schizophrenia: A 4 Tesla Proton Magnetic Resonance Spectroscopy Study. AB - This article has been retracted, consistent with Elsevier Policy on Article Withdrawal. Please see . The Publisher apologises for any inconvenience this may cause. PMID- 16476411 TI - Chronic cocaine prevents depotentiation at corticostriatal synapses. AB - BACKGROUND: The advanced stages of addiction are characterized by compulsive drug seeking and drug-taking behaviors despite the loss of the hedonic effect of drug consumption. A pathology of habit forming systems might underlie these features of addiction. METHODS: We have compared use-dependent plasticity of corticostriatal synapses in saline- and cocaine-treated rats by means of single neuron electrophysiological recordings. RESULTS: High-frequency stimulation of cortical afferents induced long-term potentiation (LTP) of corticostriatal synapses in treated and untreated animals. Saline- and acute-cocaine-treated rats, however, showed synaptic depotentiation in response to subsequent low frequency stimulation of the same pathway, whereas chronic cocaine-treated animals were refractory to this process. Depotentiation was also absent in control slices bathed with cocaine, dopamine, or with the D1 receptor agonist SKF38393. The effect of cocaine on depotentiation was prevented by D1 but not D2 dopamine receptor antagonists and was mimicked by pharmacological inhibition of cyclin-dependent kinase 5, to enhance D1-receptor-associated intracellular signaling. CONCLUSIONS: These results provide the first evidence that cocaine blocks the reversal of LTP in brain circuits. This alteration might be important for the persistence of addictive behavior despite efforts to abstain. PMID- 16476412 TI - Catechol-o-methyltransferase, cognition, and psychosis: Val158Met and beyond. AB - This review summarizes our current understanding of catechol-o-methyltransferase (COMT) and how it relates to brain function and schizophrenia. We begin by considering the COMT gene, its transcripts and proteins, and its relevance for central catecholamine function. We then describe how variation in COMT activity affects the function of the prefrontal cortex (PFC) and associated areas, reviewing evidence that COMT modulates executive function and working memory and highlighting recent data that also implicate it in emotional processing. Finally, we discuss briefly the genetic association between COMT and schizophrenia, focusing in particular on the complex interaction of functional loci within the gene that may underlie the mixed results of studies to date. We conclude by outlining preliminary data indicating that COMT is a promising therapeutic target for ameliorating the cognitive deficits associated with schizophrenia. PMID- 16476413 TI - Significant association between the genetic variations in the 5' end of the N methyl-D-aspartate receptor subunit gene GRIN1 and schizophrenia. AB - BACKGROUND: N-methyl-D-aspartate (NMDA) receptors play important roles in many neurophysiological processes. Evidence from previous studies indicate that NMDA receptors contribute to the pathophysiology of schizophrenia. Two NMDA receptor subunit genes, GRIN1 and GRIN2A, are both good candidate genes for schizophrenia. METHOD: We genotyped five single nucleotide polymorphisms (SNPs) in GRIN1 and two in GRIN2A in 2455 Han Chinese subjects, including population- and family-based samples, and performed case-control and transmission disequilibrium test (TDT) analyses. A microsatellite in GRIN2A was genotyped in population-based samples and a Mann-Whitney U test was performed. RESULTS: A highly significant association was detected at the 5' end of GRIN1. Analyses of single variants and multiple-locus haplotypes indicate that the association is mainly generated by rs11146020 (case-control study: p = .0000013, odds ratio = .61, 95% confidence interval .50-.74; TDT: p = .0019, T/NT = 79/123). No association was found in the GRIN2A polymorphisms. CONCLUSIONS: Our results provide support for the hypothesis that NMDA receptors are an important factor in schizophrenia. Moreover, rs11146020 is located in 5' untranslated region where several functional elements have been found. Hence, the SNP is a potential candidate in altering risk for schizophrenia and worthy of further replication and functional study. PMID- 16476414 TI - An event-related fMRI study of the neural networks underlying repetition suppression and reaction time priming in implicit visual memory. AB - Unfamiliar line drawings were presented to subjects three times during BOLD fMRI scanning. A set of brain areas was detected in which the effect of stimulus repetition on the evoked fMRI response depended on whether or not the drawing could be conceived as a coherent three-dimensional structure. Differential repetition effects were found in the neural response to drawings of both structurally possible and impossible objects. This differential effect of repetition was related to the amount of reaction time priming on the concurrent task involving decisions about three-dimensional structure in the possible but not in the impossible objects. These results point to different neurophysiological processing mechanisms for structurally possible and impossible images and demonstrate neural plasticity that predicts behavioral priming for structurally possible images. PMID- 16476415 TI - Enhanced expression of heat shock protein 27 is correlated with axonal regeneration in mature retinal ganglion cells. AB - The small heat shock protein, Hsp27, promotes axonal regeneration in peripheral neurons; however, an analogous role in the central nervous system has not been described. This study examined the relationship between Hsp27 expression and regeneration in mature retinal ganglion cells (RGCs). Adult rat optic nerves were transected and exposed to peripheral nerve autografts to stimulate regeneration of cut RGC axons. There was a five-fold increase in the Hsp27-positive fraction of RGCs that extended new axons into the graft when compared with those that survived injury but did not regenerate (30% versus 6% respectively, P = 0.001). Hsp27 protein was located throughout somata and neuritic processes, and there was a significant positive correlation between Hsp27 expression and axonal regeneration in injured neurons (R = 0.92, P < 0.0001). These findings are consistent with the growth-associated role of Hsp27 demonstrated in peripheral neurons and suggest that Hsp27 may mediate similar physiological functions in the central nervous system. PMID- 16476417 TI - Visual spatial integrity in the absence of splenium. AB - Several lines of evidence have confirmed that visual integrity among the two hemifields is severely impaired following transection of posterior part of the corpus callosum (splenium). But the extent of this impairment differs for various visual functions. Here, in a posterior callosectomized patient (MD), interhemispheric visual spatial integrity is investigated in three experiments. In the first experiment, MD was asked to make decision about horizontal alignment of two balls presented simultaneously in one of three conditions: both in right or left hemifield, or each in one hemifield. We have previously shown that she is not able to perform a shape-matching task for stimuli presented to different hemifields. But in this task, MD was able to compare location of the two bilaterally presented stimuli significantly above chance level. Then we investigated whether attentive visual object tracking across vertical meridian of the visual field is possible in the absence of splenium. MD had to attentively track one bouncing ball among three identical balls, while it crossed the vertical midline in half of trials. Her performance in crossed conditions was significantly above chance level, but it was lower than uncrossed conditions. Finally, we investigated the contribution of simple interhemispheric temporal signals in performing the attentive tracking task. Results suggest that the patient was not using such temporal signals. Our results suggest that interhemispheric connections other than splenium can contribute in making an integral visual map across hemifields. Such an integrated map can be used for bilateral visual spatial comparisons and visual spatial attention. PMID- 16476416 TI - mu- but not delta- and kappa-opioid receptors in the ventrolateral orbital cortex mediate opioid-induced antiallodynia in a rat neuropathic pain model. AB - Previous studies have indicated that the ventrolateral orbital cortex (VLO) is involved in opioid-mediated antinociception in the tail flick test and formalin test. The aim of the current study was to examine the effect of opioids microinjected into the VLO on allodynia in the rat L5/L6 spinal nerve ligation (SNL) model of neuropathic pain and determine the roles of different subtypes of opioid receptors in this effect. The allodynia was assessed by both mechanical (von Frey filaments) and cold plate (4 degrees C) stimuli. Morphine (1.0, 2.5, and 5.0 microg) microinjected into the VLO contralateral to the nerve ligation dose-dependently depressed the mechanical and cold allodynia and these effects were reversed by nonselective opioid receptor antagonist naloxone (1.0 microg) administrated into the same site. Microinjection of endomorphin-1 (5.0 microg), a highly selective mu-opioid receptor agonist, and [D-Ala2, D-Leu5]-enkephalin (DADLE, 10 microg), a delta-/mu-opioid receptor agonist, also depressed the allodynia, and the effects of both drugs were blocked by selective mu-receptor antagonist beta-funaltrexamine (beta-FNA, 3.75 microg), but the effects of DADLE were not influenced by the selective delta-receptor antagonist naltrindole (5.0 microg). Microinjection of U-62066 (100 microg), a kappa-opioid receptor agonist, into the VLO had no effect on the allodynia. These results suggest that the VLO is involved in opioid-induced antiallodynia and mu- but not delta- and kappa opioid receptor mediates these effects in the rat with neuropathic pain. PMID- 16476418 TI - Age-related differences in response regulation as revealed by functional MRI. AB - This fMRI study studied age-related differences in neural activities during response regulation. Twenty-one male participants from two age groups, a younger group and an older group (mean ages: 29.9 and 65.2 years, respectively), were scanned while performing a task with response compatibility manipulation. They were presented with a sequence of arrowheads that pointed either upward or downward. In the "Response Compatible" condition, they were required to press an up or a down button consistent with the direction of the arrowhead. In the "Response Incompatible" condition, they were required to press the button opposite to the arrowhead direction so that an upward arrow should elicit a down response, and vice versa. Findings showed age-related differences in response regulation in several brain regions, including the right frontal, the right cingulate, and the left inferior parietal cortexes. The findings suggested a higher level of neural activity in the right prefrontal and left inferior parietal regions during response regulation for the older adults than for the younger adults. PMID- 16476419 TI - Defensive-like behaviors and antinociception induced by NMDA injection into the periaqueductal gray of mice depend on nitric oxide synthesis. AB - Glutamate NMDA receptor activation within the periaqueductal gray (PAG) leads to antinociceptive, autonomic and behavioral responses characterized as the fear reaction. Considering that NMDA receptor triggers activation of neuronal nitric oxide synthase (nNOS), enzyme that produces nitric oxide (NO), this study investigated the effects of intra-PAG infusions of NPLA (Nomega-propyl-L arginine), an nNOS inhibitor, on behavioral and antinociceptive responses induced by local injection of NMDA receptor agonist in mice. The behaviors measured were frequency of jumping and rearing as well as duration (in seconds) of running and freezing. Nociception was assessed during the second phase of the formalin test (injection of 50 microl of formalin 2.5% into the dorsal surface of the right hind paw). Five to seven days after stereotaxic surgery for intracerebral cannula implantation, mice were injected with formalin into the paw, and 10 min later, they received intra-dPAG injection of NPLA (0, 0.2, or 0.4 nmol/0.1 microl). Ten minutes later, they were injected with NMDA (N-methyl-D-aspartate: 0 or 0.04 nmol/0.1 microl) into the same midbrain site and were immediately placed in glass holding cage for recording the defensive behavior and the time spent on licking the injected paw with formalin during a period of 10 min. Microinjections of NMDA significantly decreased nociception response and produced jumping, running, and freezing reactions. Intra-dPAG injections of NPLA (0.4 nmol) completely blocked the NMDA effects without affecting either behavioral or nociceptive responses in intra-dPAG saline-injected animals, except for the rearing frequency that was increased by the nNOS inhibitor. These results strongly suggest the involvement of NO within the PAG in the antinociceptive and defensive reactions induced by local glutamate NMDA receptor activation in this midbrain structure. PMID- 16476420 TI - Responses during cell preparation for functional analyses in mouse bone marrow derived cultured mast cells. AB - Murine bone marrow-derived cultured mast cells (BMMCs) are most widely used in in vitro experiments for evaluation of mast cell functions. The present study has shown that cell preparation procedure, i.e., cell collection by centrifugation and the subsequent adjustment and culture of cell density at the desired concentrations, transiently induced gene expression of plasminogen activator inhibitor-1 (PAI-1) and the AP-1 components (c-fos, c-jun, and junB). The level of PAI-1 gene transcript was closely related to the cell density and the gene expression was enhanced by pretreatment with okadaic acid, an inhibitor of protein phosphatases 1 (PP1) and 2A (PP2A). The cell preparation procedure also caused dephosphorylation of MAP kinases, i.e., ERK, p38, and JNK, resulting from PP1/PP2A activation. In view of the cell responses to the cell preparation procedure itself, care is needed in the interpretation of in vitro data using BMMCs. PMID- 16476421 TI - Selection of optimal AR spectral estimation method for EEG signals using Cramer Rao bound. AB - Electroencephalography is an essential clinical tool for the evaluation and treatment of neurophysiologic disorders related to epilepsy. Careful analyses of the electroencephalograph (EEG) records can provide valuable insight and improved understanding of the mechanisms causing epileptic disorders. The detection of epileptiform discharges in the EEG is an important element in the diagnosis of epilepsy. In this study, EEG signals recorded from 30 subjects were processed using autoregressive (AR) method and EEG power spectra were obtained. The parameters of autoregressive method were estimated by different methods such as Yule-Walker, covariance, modified covariance, Burg, least squares, and maximum likelihood estimation (MLE). EEG spectra were then used to analyze and characterize epileptiform discharges in the form of 3-Hz spike and wave complexes in patients with absence seizures. The variations in the shape of the EEG power spectra were examined in order to obtain medical information. These power spectra were then used to compare the applied methods in terms of their frequency resolution and determination of epileptic seizure. The Cramer-Rao bounds (CRB) were derived for the estimated AR parameters of the EEG signals and the performance evaluation of the estimation methods was performed using the CRB values. Finally, the optimal AR spectral estimation method for the EEG signals was selected according to the computed CRB values. According to the computed CRB values, the performance characteristics of the MLE AR method was found extremely valuable in EEG signal analysis. PMID- 16476422 TI - PTH/PTHrP receptor delays chondrocyte hypertrophy via both Runx2-dependent and independent pathways. AB - The transcription factor, Runx2, promotes chondrocyte hypertrophy, whereas parathyroid hormone-related protein (PTHrP) delays this process. To examine whether PTHrP suppresses chondrocyte hypertrophy via Runx2-dependent or independent pathways, Runx2 expression and chondrocyte differentiation were analyzed using bones from embryonic limbs of wild type and Runx2(-/-) mice. Treatment of cultured rudiments with PTH dramatically suppresses Runx2 mRNA levels in hypertrophic chondrocytes. PTH-induced delay of chondrocyte hypertrophy was observed in cultured tibiae from both Runx2(-/-) and wild-type embryos. This delay was also seen after PTH administration to limbs from wild type and Runx2(-/ ) mice expressing Runx2 in chondrocytes via a collagen 2 promoter-driven transgene. To further explore Runx2-dependent and -independent effects of PTHrP, we examined embryonic tibiae and femurs from littermates null for PTHrP, Runx2, or both genes. Runx2(-/-) femurs exhibited no vascular invasion or chondrocytes expressing collagen type X or osteopontin mRNA. In contrast, Runx2(-/-)/PTHrP(-/ ) mice exhibited limited vascular invasion and some chondrocytes expressing collagen X or osteopontin mRNA. In both tibia and femur, Runx2(-/-)/PTHrP(-/-) mice exhibited expanded regions of proliferating chondrocytes when compared to the same regions in PTHrP(-/-) mice. These data indicate that the delayed hypertrophy induced by PTHrP is mediated by both Runx2-dependent and -independent mechanisms. PMID- 16476423 TI - Molecular mechanisms of axon guidance. AB - In order to form a functional nervous system, neurones extend axons, often over long distances, to reach their targets. This process is controlled by extracellular receptors and their ligands, several families of which have been identified. These proteins may act to either repel or attract growth cones and a given receptor may transduce either type of signal, depending on the cellular context. In addition to these archetypal axon guidance molecules, it is becoming apparent that molecules previously known for their role in patterning can also direct axonal outgrowth. The growth cone receptors do not act in isolation and combine with members of the same or other families to produce a graded response or even a complete reversal in its polarity. These signals can be further combined and/or modulated by processing of the molecule both directly at the cell surface and by the network of intracellular signalling pathways which are activated. The result is a sophisticated and dynamic set of cues that enable a growth cone to successfully navigate to its destination, modulating its response to changing environmental cues along its pathway. PMID- 16476424 TI - Curcumin inhibits the proliferation and mineralization of cultured osteoblasts. AB - The effects of curcumin, which is an important constituent of rhizomes of the plant Curcuma longa Linn, on the metabolism of osteoblasts were examined in cultures of rat calvarial osteoblastic cells (ROB cells). The proliferation of cells was markedly inhibited upon exposure of cells to curcumin at 5x10(-6) to 1x10(-5) M. Curcumin at 1x10(-5) M did not induce apoptosis in ROB cells but arrested cells at the G1 phase of the cell cycle. In addition, curcumin stimulated the expression of mRNA for p21(WAF1/CIP1), which inhibits the activity of cyclin-dependent kinases, and inhibited the phosphorylation of histone H1. Furthermore, curcumin reduced the rate of deposition of calcium and the formation of mineralized nodules. Our results indicate that curcumin might inhibit the proliferation and mineralization of osteoblastic cells through the expression of p21(WAF1/CIP1). PMID- 16476425 TI - Zasp/Cypher internal ZM-motif containing fragments are sufficient to co-localize with alpha-actinin--analysis of patient mutations. AB - Z-band alternatively spliced PDZ-containing protein (ZASP/Cypher) has an important role in maintaining Z-disc stability in striated and cardiac muscle. ZASP/Cypher interacts through its PDZ domain with the major Z-disc actin cross linker, alpha-actinin. ZASP/Cypher also has a conserved sequence called the ZM motif, and it is found in two alternatively spliced exons 4 and 6. We have shown earlier that the ZM-motif containing internal regions of two related proteins ALP and CLP36 interact with alpha-actinin rod region, and that the ZM-motif is important in targeting ALP to the alpha-actinin containing structures in cell. Here, we show that the ZASP/Cypher internal fragments containing either ZM exon 4 or 6 co-localized with alpha-actinin in cultured myoblasts and nonmuscle cells. Fragments of 130 residues around the ZM-consensus were sufficient for localization, which is similar to our previous results of ALP. Moreover, ZASP/Cypher protein interacted directly with the alpha-actinin rod and competed with ALP in binding to the rod. During the inhibition of stress fiber assembly ZASP/Cypher and alpha-actinin co-localization could be partially disturbed, suggesting that ZASP/Cypher is bound to alpha-actinin mainly when alpha-actinin is localizing in stress fibers. Many point mutations found in cardiomyopathy patients are located in the internal region of ZASP/Cypher. However, we found no evidence that human patient mutations in the internal domain would affect the ZASP/Cypher co-localization with alpha-actinin, or that the mutations would destabilize the ZASP/Cypher protein. PMID- 16476426 TI - pat-4/ILK and unc-112/Mig-2 are required for gonad function in Caenorhabditis elegans. AB - Tissue morphogenesis requires proper interaction between cells and the extracellular matrix (ECM), which is mediated by alphabeta heterodimeric receptor integrin. In Caenorhabditis elegans, integrin signaling is essential for formation of gonad. Here, we probe the role of several integrin-associated molecules in ovulation and cell migration. Function of pat-4/integrin-linked kinase (ILK) and unc-112/Mig-2 was examined using RNA-mediated interference (RNAi). Depletion of these messages caused oocyte accumulation in the proximal gonad and distal tip cells (DTC) migration defects. It was further determined that failed ovulation was due to defective contraction and dilation of somatic gonad structures, including spermatheca and gonad sheath. Actin cytoskeleton in the proximal gonad of RNAi animals appeared disorganized, indicating that RNAi of pat-4 or unc-112 inhibited the overall assembly of actin cytoskeleton in somatic gonad. Taken together, our analysis confirms the role of integrin and integrin associated proteins in gonad function. PMID- 16476427 TI - First look at the effect of overexpression of TIGR/MYOC on the transcriptome of the human trabecular meshwork. AB - Wild-type TIGR/MYOC is a secreted protein implicated in the development of steroid glaucoma. Mutations in TIGR/MYOC have been linked to some patients who develop elevated intraocular pressure (IOP) and glaucoma. Because there is evidence of some other factors contributing to the TIGR/MYOC causative role in glaucoma, and because substantial increased levels of a particular cellular mRNA and protein might alter expression of other host genes, we began to investigate the effect of TIGR/MYOC overexpression on the transcriptome of human trabecular meshwork cells. We used a recombinant adenovirus carrying wild-type TIGR/MYOC cDNA, primary HTM cells, 300 viral particles per cell and U133 Affymetrix GeneChips. Our results indicate that 2361 out of the 22,284 genes (10.6%) were altered more than two-fold (p55 years were at significant risk to receive increased transfusions (P < .001 and P = .035, respectively). An admission Hb < or =12 g/dL and any mention of long bone orthopedic operations or laparotomy or thoracotomy were associated with increased risk of blood transfusion during the first week of admission. Logistic regression analysis identified transfusion of >4 units of blood as a significant risk factor for SIRS. After 1 week of ICU stay, ISS > 20 and blunt injury were associated with increased risk of transfusion. CONCLUSIONS: Trauma patients are heavily transfused with allogeneic blood throughout the course of their hospital stay and transfusions are administered at relatively high pre-transfusion haemoglobin levels (mean of 9 g/dL). Transfusion of >4 units of blood is an independent risk factor for SIRS. Strategies to limit blood transfusions should be investigated in this population. PMID- 16476430 TI - Spontaneous odontoid process fracture in rheumatoid arthritis: diagnostic difficulties, pathology and treatment. PMID- 16476431 TI - A randomised study in all cervical hip fractures osteosynthesis with Hansson hook pins versus AO-screws in 199 consecutive patients followed for two years. AB - A consecutive series of patients with all types of cervical hip fracture (both undisplaced and displaced) were randomised to osteosynthesis with Hansson hook pins (n = 98) or AO-screws (n = 101). Background parameters, fracture type and reduction of the fracture did not differ significantly between the groups. Fifty seven percent of the patients were operated on within 6 h of admission to hospital, 74% within 12 h and 92% within 24 h. The mean (median) time for operation was 36 (30) min for the hook-pins and 40 (35) min for the AO-screws. The devices were significantly better positioned in the hook-pin group (81% of cases good) compared to the AO-screws (66% good) (p = 0.04). In all, 72% of the patients had no deficiency either in reduction of the fracture, positioning of the implants or had drill penetration of the femoral head. Direct unrestricted weight bearing was encouraged in 92% of the hook-pin and 90% of the AO-screws group. The mean (median) hospital time was 13 (10) days with no significant difference between the groups. Following treatment, 5% walked without aids, 76% of the patients walked with some aids, and 16% could not walk. The walking ability was not known for 4%. At four months, 59% of the patients were living in their own home (64% before fracture), 18% (25% before) in a nursing home, 5% (11% before) in other accommodation and 18% were dead. After two years, 77% of the hook-pin patients had not needed any re-operation compared to 73% in the AO-screw group. In total a secondary hemi-arthroplasty had been performed in 7% and total hip arthroplasty in 12% of the patients. Extraction only of osteosynthesis material had been performed in 5%. The difference in the reoperation rates between the two methods was not significant. In the undisplaced fractures, 84% of the patients had not needed any reoperation after two years compared to 70% among the displaced fractures. Major reoperation had been performed in 10% (1% hemi and 9% total hip arthroplasty) in the patients with undisplaced fractures compared to 26% in those with displaced fractures (10% hemi, 16% total hip arthroplasty and 1% Girdlestone operation). The remaining patients had only undergone removal of metalwork. Osteosynthesis thus proved to be a successful operation in many of the patients with displaced fractures. A preoperative, prognostic-based selection between osteosynthesis and arthroplasty is the future goal for optimised femoral neck fracture treatment. PMID- 16476432 TI - Work stress among nursing home care attendants in Taiwan: a questionnaire survey. AB - BACKGROUND: Care attendants constitute the main workforce in nursing homes, but their heavy workload, low autonomy, and indefinite responsibility result in high levels of stress and may affect quality of care. However, few studies have focused of this problem. OBJECTIVES: The aim of this study was to examine work related stress and associated factors that affect care attendants in nursing homes and to offer suggestions for how management can alleviate these problems in care facilities. METHODS: We recruited participants from nine nursing homes with 50 or more beds located in middle Taiwan; 110 care attendants completed the questionnaire. The work stress scale for the care attendants was validated and achieved good reliability (Cronbach's alpha=0.93). We also conducted exploratory factor analysis. RESULTS: Six factors were extracted from the work stress scale: insufficient ability, stressful reactions, heavy workload, trouble in care work, poor management, and working time problems. The explained variance achieved 64.96%. Factors related to higher work stress included working in a hospital based nursing home, having a fixed schedule, night work, feeling burden, inconvenient facility, less enthusiasm, and self-rated higher stress. CONCLUSION: Work stress for care attendants in nursing homes is related to human resource management and quality of care. We suggest potential management strategies to alleviate work stress for these workers. PMID- 16476433 TI - Toll-like receptor recognition of Toxoplasma gondii. AB - Toxoplasma gondii potently stimulates IFN-gamma production by both the innate and adaptive immune system as part of its host adaptation. This response is known to be dependent on an Myeloid Differentiation factor 88 signaling pathway used by Toll-like receptors (TLRs), a family of proteins involved in the recognition of microbial molecular patterns. In the following review, we summarise the evidence for specific TLR function in host resistance to T. gondii focusing on the recent discovery in the parasite of a profilin-like ligand that potently stimulates TLR11 and regulates the production of IL-12, a cytokine necessary for the protective IFN-gamma response. In addition, we discuss the hypothesis that TLR11 may have evolved as a general pattern recognition receptor for apicomplexan protozoa and that as highly conserved proteins associated with actin-based motility, profilins are logical ligand targets for this form of pathogen detection. Finally, we review the evidence for involvement of other TLR and TLR ligands in host resistance to T. gondii and discuss how such receptors might synergise with TLR11 in the innate response to the parasite. PMID- 16476434 TI - Autoantibodies to oxidized LDL and cardiovascular risk: the Framingham Offspring Study. AB - BACKGROUND: The relation between measures of oxidation of lipid particles and cardiovascular disease has not been extensively investigated prospectively on a population basis. METHODS: A community cohort of 1192 men and 1427 women with measures of IgG antibodies to oxidized LDL were followed 8 years for the development of initial coronary heart disease (CHD) and cardiovascular disease (CVD) events. RESULTS: Levels of IgG autoantibodies to a form of oxidized LDL were significantly associated with age in both sexes, positively with fibrinogen in men and negatively with HDL cholesterol in women. In sex-specific models that adjusted for age alone or those that adjusted for age, cholesterol, HDL cholesterol, smoking, and diabetes mellitus, there was no relation between level of antibodies to oxidized LDL and the development of CHD or CVD. CONCLUSION: Autoantibodies to oxidized LDL were strongly related to age and were not related to incident CHD or CVD over 8 years of follow up. PMID- 16476435 TI - High-performance liquid chromatographic enantioseparations on capillary columns containing monolithic silica modified with amylose tris(3,5 dimethylphenylcarbamate). AB - Monolithic capillary columns containing native silica were modified by in situ coating with amylose tris(3,5-dimethylphenylcarbamate) and applied for enantioseparations in capillary liquid chromatography. Capillary columns were examined for 10 standard racemic compounds in order to compare the performance of monolithic silica columns with the common, 4.6mm I.D. high-performance liquid chromatographic columns packed with particulate silica. The effects of polysaccharide coating and of the linear velocity of the mobile phase on peak performance were studied. Enantioseparations with an analysis time below 1min were achieved for some chiral analytes. PMID- 16476436 TI - Ultrafast concentration and speciation of chromium(III) and (VI). AB - There is an increasing need to know the concentrations of chromium(III) and (VI) separately rather than only the total chromium content. A method is described for accomplishing this very quickly using only low-cost, portable equipment. Two small, resin-loaded extraction disks are placed one on top of the other in a plastic holder. Then a syringe containing the aqueous sample is attached to the holder and the sample is pushed through the disks. In a matter of seconds, all of the chromium(VI) is retained on the top anion-exchange disk and chromium(III) is extracted by the second cation-exchange disk. The concentrations on each disk are several hundredfold higher than they were in the original sample. The amounts of chromium(III) and (VI) extracted are measured directly on the surface of the respective disks by diffuse reflectance spectroscopy (DRS). Despite the low molar absorptivity of chromium(III) in aqueous solution, the concentration on the upper most layer on the extraction disk is high enough to permit the determination of chromium(III) in samples at the low mg/L range. Chromium(VI) can also be determined at low to sub-mg/L concentrations. A study of the cation-exchange disks was undertaken to compare the performance characteristics of disks containing sulfonated resins and those with iminodiacetate functionality. In addition, data are presented to show the effects of heating the iminodiacetate disks after the initial extraction. The disks were heated in hot water for 15-30 min to complete the slow complexation reaction on the surface. PMID- 16476437 TI - Influence of frictional heating on temperature gradients in ultra-high-pressure liquid chromatography on 2.1mm I.D. columns. AB - The effects of viscous heat dissipation on some important HPLC parameters, such as efficiency (N) and retention factors (k), using 2.1mm columns at pressures up to 1000 bar have been investigated from both a theoretical and experimental point of view. Two distinct experimental set-ups and their respective influences on non homogenous temperature gradients within the column are described and discussed. In the first instance, a still-air column heater was used. This set-up leads to approximate 'adiabatic' conditions, and a longitudinal temperature gradient is predicted across the length of the column. The magnitude of this gradient is calculated, and its occurrence confirmed with experimental measurements also indicating that no appreciable loss in efficiency occurs. Secondly, when a water bath is used to thermostat the column, a radial temperature gradient is prevalent. The extent of this gradient is estimated, and the loss in efficiency associated with this gradient is predicted and demonstrated experimentally. It is also observed that approximate adiabatic conditions can lead to floating retention factors. The implications of temperature gradients for routine HPLC analysis at ultra-high pressure are discussed. PMID- 16476438 TI - Fast determination of anions on a short coated column. AB - In this paper, a simple method for the separation and determination of common inorganic anions by fast ion-exchange chromatography, using a modified short (25 mm x 4.6 mm) monolithic column, is reported. Coating the column with a cationic surfactant, cetylpyridinium chloride (CPC), the isocratic separation of some inorganic anions in minutes was possible, by direct or indirect UV detection. The coated column demonstrated excellent stability over time, even at a high flow rate, giving retention times with an average relative standard deviation of 1.3% for over 10 consecutive runs. The developed column exhibited unusual selectivity for common anions, was successfully applied to the rapid analysis of inorganic anions of food samples, river water and factory waste water samples. PMID- 16476439 TI - Molecular mechanisms of PKCalpha localization and activation by arachidonic acid. The C2 domain also plays a role. AB - Arachidonic acid, one of the major unsaturated fatty acids released during cell stimulation, participates in the signaling necessary for activation of different enzymes, including protein kinase C (PKC). Here, we demonstrate that arachidonic acid is a direct activator of PKCalpha, but needs the cooperation of Ca(2+) to exert its function. By using several mutants of the C2 and C1 domains, we were able to determine the molecular mechanism of this activation. More specifically, site-directed mutagenesis in key residues found in the C2 domain showed that the Ca(2+)-binding region was essential for the arachidonic acid-dependent localization and activation of PKCalpha. However, the lysine-rich cluster, also located in the C2 domain, played no relevant role in either the membrane localization or activation of the enzyme. Moreover, site-directed mutagenesis in key residues placed in the C1A and C1B subdomains, which are responsible for the diacylglycerol/phorbil ester interaction, demonstrated that the C1A subdomain was involved in the membrane localization and activation mechanism. Taken together, these data suggest a very precise mechanism for PKCalpha activation by arachidonic acid, involving a sequential model of activation in which an increase in intracytosolic Ca(2+) leads to the interaction of arachidonic acid with the Ca(2+)-binding region; only after this step, does the C1A subdomain interact with arachidonic acid, leading to full activation of the enzyme. PMID- 16476440 TI - Effects of calcium binding on the side-chain methyl dynamics of calbindin D9k: a 2H NMR relaxation study. AB - The effects of Ca(2+) binding on the side-chain methyl dynamics of calbindin D(9k) have been characterized by (2)H NMR relaxation rate measurements. Longitudinal, transverse in-phase, quadrupolar order, transverse anti-phase and double quantum relaxation rates are reported for both the apo and Ca(2+)-loaded states of the protein at two magnetic field strengths. The relatively large size of the data set allows for a detailed analysis of the underlying conformational dynamics by spectral density mapping and model-free fitting procedures. The results reveal a correlation between a methyl group's distance from the Ca(2+) binding sites and its conformational dynamics. Several methyl groups segregate into two limiting classes, one proximal and the other distal to the binding sites. Methyl groups in these two classes respond differently to Ca(2+) binding, both in terms of the timescale and amplitude of their fluctuations. Ca(2+) binding elicits a partial immobilization among methyl groups in the proximal class, which is consistent with previous studies of calbindin's backbone dynamics. The distal class, however, exhibits a trend that could not be inferred from the backbone data in that its mobility actually increases with Ca(2+) binding. We have introduced the term polar dynamics to describe this type of organization across the molecule. The trend may represent an important mechanism by which calbindin D(9k) achieves high affinity binding while minimizing the corresponding loss of conformational entropy. PMID- 16476441 TI - NMR structure of the WIF domain of the human Wnt-inhibitory factor-1. AB - The human Wnt-binding protein Wnt-inhibitory factor-1 (WIF-1) comprises an N terminal WIF module followed by five EGF-like repeats. Here we report the three dimensional structure of the WIF domain of WIF-1 determined by NMR spectroscopy. The fold consists of an eight-stranded beta-sandwich reminiscent of the immunoglobulin fold. Residual detergent (Brij-35) used in the refolding protocol was found to bind tightly to the WIF domain. The binding site was identified by intermolecular nuclear Overhauser effects observed between the WIF domain and the alkyl chain of the detergent. The results point to a possible role of WIF domains as a recognition motif of Wnt and Drosophila Hedgehog proteins that are activated by palmitoylation. PMID- 16476442 TI - Crystal structure of the N domain of human somatic angiotensin I-converting enzyme provides a structural basis for domain-specific inhibitor design. AB - Human somatic angiotensin I-converting enzyme (sACE) is a key regulator of blood pressure and an important drug target for combating cardiovascular and renal disease. sACE comprises two homologous metallopeptidase domains, N and C, joined by an inter-domain linker. Both domains are capable of cleaving the two hemoregulatory peptides angiotensin I and bradykinin, but differ in their affinities for a range of other substrates and inhibitors. Previously we determined the structure of testis ACE (C domain); here we present the crystal structure of the N domain of sACE (both in the presence and absence of the antihypertensive drug lisinopril) in order to aid the understanding of how these two domains differ in specificity and function. In addition, the structure of most of the inter-domain linker allows us to propose relative domain positions for sACE that may contribute to the domain cooperativity. The structure now provides a platform for the design of "domain-specific" second-generation ACE inhibitors. PMID- 16476443 TI - Prediction of binding sites of peptide recognition domains: an application on Grb2 and SAP SH2 domains. AB - Determination of the binding motif and identification of interaction partners of the modular domains such as SH2 domains can enhance our understanding of the regulatory mechanism of protein-protein interactions. We propose here a new computational method to achieve this goal by integrating the orthogonal information obtained from binding free energy estimation and peptide sequence analysis. We performed a proof-of-concept study on the SH2 domains of SAP and Grb2 proteins. The method involves the following steps: (1) estimating the binding free energy of a set of randomly selected peptides along with a sample of known binders; (2) clustering all these peptides using sequence and energy characteristics; (3) extracting a sequence motif, which is represented by a hidden Markov model (HMM), from the cluster of peptides containing the sample of known binders; and (4) scanning the human proteome to identify binding sites of the domain. The binding motifs of the SAP and Grb2 SH2 domains derived by the method agree well with those determined through experimental studies. Using the derived binding motifs, we have predicted new possible interaction partners for the Grb2 and SAP SH2 domains as well as possible interaction sites for interaction partners already known. We also suggested novel roles for the proteins by reviewing their top interaction candidates. PMID- 16476444 TI - Interactions of the release factor RF1 with the ribosome as revealed by cryo-EM. AB - In eubacteria, termination of translation is signaled by any one of the stop codons UAA, UAG, and UGA moving into the ribosomal A site. Two release factors, RF1 and RF2, recognize and bind to the stop codons with different affinities and trigger the hydrolysis of the ester bond that links the polypeptide with the P site tRNA. Cryo-electron microscopy (cryo-EM) results obtained in this study show that ribosome-bound RF1 is in an open conformation, unlike the closed conformation observed in the crystal structure of the free factor, allowing its simultaneous access to both the decoding center and the peptidyl-transferase center. These results are similar to those obtained for RF2, but there is an important difference in how the factors bind to protein L11, which forms part of the GTPase-associated center of the large ribosomal subunit. The difference in the binding position, C-terminal domain for RF2 versus N-terminal domain for RF1, explains a body of L11 mutation studies that revealed differential effects on the activity of the two factors. Very recent data obtained with small-angle X-ray scattering now reveal that the solution structure of RF1 is open, as here seen on the ribosome by cryo-EM, and not closed, as seen in the crystal. PMID- 16476445 TI - Alanine scanning mutagenesis of Abeta(1-40) amyloid fibril stability. AB - We describe here an alanine scanning mutational analysis of the Abeta(1-40) amyloid fibril monitored by fibril elongation thermodynamics derived from critical concentration values for fibril growth. Alanine replacement of most residues in the amyloid core region, residues 15-36, leads to destabilization of the elongation step, compared to wild-type, by about 1kcal/mol, consistent with a major role for hydrophobic packing in Abeta(1-40) fibril assembly. Where comparisons are possible, the destabilizing effects of Ala replacements are generally in very good agreement with the effects of Ala replacements of the same amino acid residues in an element of parallel beta-sheet in the small, globular protein Gbeta1. We utilize these Ala-WT DeltaDeltaG values to filter previously described Pro-WT DeltaDeltaG values, creating Pro-Ala DeltaDeltaG values that specifically assess the sensitivity of a sequence position, in the structural context of the Abeta fibril, to replacement by proline. The results provide a conservative view of the energetics of Abeta(1-40) fibril structure, indicating that positions 18-21, 25-26, and 32-33 within amyloid structure are particularly sensitive to the main-chain disrupting effects of Pro replacements. In contrast, residues 14-17, 22, 24, 27-31, and 34-39 are relatively insensitive to Pro replacements; most N-terminal residues were not tested. The results are discussed in terms of amyloid fibril structure and folding energetics, in particular focusing on how the data compare to those from other structural studies of Abeta(1-40) amyloid fibrils grown in phosphate-buffered saline at 37 degrees C under unstirred ("quiescent") conditions. PMID- 16476446 TI - Building a virus from scratch: assembly of an infectious virus using purified components in a rigorously defined biochemical assay system. AB - The assembly of double-stranded DNA (dsDNA) viruses such as poxvirus, the herpesviruses and many bacteriophages is a complex process that requires the coordinated activities of numerous proteins of both viral and host origin. Here, we report the assembly of an infectious wild-type lambda virus using purified proteins and commercially available DNA, and optimization of the assembly reaction in a rigorously defined biochemical system. Seven proteins, purified procapsids and tails, and mature lambda DNA are necessary and sufficient for efficient virus assembly in vitro. Analysis of the reaction suggests that (i) virus assembly in vitro is optimal under conditions that faithfully mimic the intracellular environment within an Escherichia coli cell, (ii) concatemeric DNA is required for the successful completion of virus assembly, (iii) several of the protein components oligomerize concomitant with their step-wise addition to the nascent virus particle and (iv) tail addition is the rate-limiting step in virus assembly. Importantly, the assembled virus may enter either of the developmental pathways (lytic or lysogenic) expected of a lambda virion. Thus, we demonstrate for the first time that a wild-type, complex DNA virus may be assembled from purified components under defined biochemical conditions. This system provides a powerful tool to characterize, at the molecular level, the step-by-step processes required to assemble an infectious virus particle. Given the remarkable similarities between dsDNA bacteriophage and eukaryotic dsDNA viruses, characterization of the lambda system has broad biological implications in our understanding of virus development at a global level. PMID- 16476447 TI - K644E/M FGFR3 mutants activate Erk1/2 from the endoplasmic reticulum through FRS2 alpha and PLC gamma-independent pathways. AB - Fibroblast growth factor receptors 3 (FGFR3) with K644M/E substitutions are associated to the severe skeletal dysplasias: severe achondroplasia with developmental delay and achanthosis nigricans(SADDAN) and thanatophoric dysplasia(TDII). The high levels of kinase activity of the FGFR3-mutants cause uncompleted biosynthesis that results in the accumulation of the immature/mannose rich, phosphorylated receptors in the endoplasmic reticulum (ER) and STATs activation. Here we report that FGFR3 mutants activate Erk1/2 from the ER through an FRS2-independent pathway: instead, a multimeric complex by directly recruiting PLCgamma, Pyk2 and JAK1 is formed. The Erk1/2 activation from the ER however, is PLCgamma-independent, since preventing the PLCgamma/FGFR3 interaction by the Y754F substitution does not inhibit Erks. Furthermore, Erk1/2 activation is abrogated upon treatment with the Src inhibitor PP2, suggesting a role played by a Src family member in the pathway from the ER. Finally we show that the intrinsic kinase activity by mutant receptors is required to allow signaling from the ER. Overall these results highlight how activated FGFR3 exhibits signaling activity in the early phase of its biosynthesis and how segregation in a sub cellular compartment can affect the FGFR3 multi-faceted capacity to recruit specific substrates. PMID- 16476448 TI - Novel inhibitors of hepatitis C virus RNA-dependent RNA polymerases. AB - Hepatitis C virus (HCV) is a major cause of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma worldwide-and is the main cause of adult liver transplants in developed nations. We have identified a class of novel and specific inhibitors of HCV NS5B RNA-dependent RNA polymerase (RdRp) activity in vitro. Characterization of two such inhibitors, COMPOUND1 (5-(4 chlorophenylmethylene)-3-(benzenesulfonylamino)-4-oxxo-2-thionothiazolidine) and COMPOUND2 (5-(4-bromophenylmethylene)-3-(benzenesulfonylamino)-4-oxxo-2 thionothiazolidine), is reported here. With IC(50) values of 0.54muM and 0.44muM, respectively, they are reversible and non-competitive with nucleotides. Biochemical and structural studies have suggested that these compounds can inhibit the initiation of the RdRp reaction. Interestingly, these inhibitors appear to form a reversible covalent bond with the NS5B cysteine 366, a residue that is not only conserved among all HCV genotypes and a large family of viruses but also required for full NS5B RdRp activity. This may reduce the potential resistance of the viruses to this class of inhibitors. PMID- 16476449 TI - Bilateral movement training and stroke rehabilitation: a systematic review and meta-analysis. AB - OBJECTIVE AND DESIGN: Bilateral movement training is being increasingly used as a post-stroke motor rehabilitation protocol. The contemporary emphasis on evidence based medicine warrants a prospective meta-analysis to determine the overall effectiveness of rehabilitating with bilateral movements. METHODS: After searching reference lists of bilateral motor recovery articles as well as PubMed and Cochrane databases, 11 stroke rehabilitation studies qualified for this systematic review. An essential requirement for inclusion was that the bilateral training protocols involved either functional tasks or repetitive arm movements. Each study had one of three common arm and hand functional outcome measures: Fugl Meyer, Box and Block, and kinematic performance. RESULTS: The fixed effects model primary meta-analysis revealed an overall effect size (ES=0.732, S.D.=0.13). These findings indicate that bilateral movement training was beneficial for improving motor recovery post-stroke. Moreover, a fail-safe analysis indicated that 48 null effects would be necessary to lower the mean effect size to an insignificant level. CONCLUSION: These meta-analysis findings indicate that bilateral movements alone or in combination with auxiliary sensory feedback are effective stroke rehabilitation protocols during the sub-acute and chronic phases of recovery. PMID- 16476450 TI - The effects of Guillain-Barre syndrome on the close relatives of patients during the first year. AB - OBJECTIVE: To study the impact of Guillain-Barre Syndrome (GBS) on the psychosocial functioning of the closest relative and on family functioning during the first year after GBS. METHOD: At 1 (=T1), 3 (=T3), 6 (=T6), and 12 months (=T12) after the onset of GBS, relatives of patients received the General Health Questionnaire (GHQ28) and the Family Assessment Device (FAD). Sixty-three relatives returned the GHQ28 at all four designated intervals. At T1 the relatives also received a questionnaire that contained questions on the impact on their daily life. The answers to these questions yielded a Daily Living Impact index. From the 110 relatives, 86 returned this questionnaire. RESULTS: 72% of the 86 relatives reported one or more problems in daily living. At T1 the scores of the GHQ subscales ranged from normal to mildly disturbed. The relatives showed significant improvement in their somatic complaints and anxiety during the first half year. Social dysfunction remained somewhat less than normal, severe depression was not found. At T1 and T3 the scores of the GHQ28 and some subscales differed significantly depending on the severity of the functional status of the patient, but not at T6 and T12. Relatives of patients with severe residua at 1 month score worse on the GHQ28 and most subscales at 6 months. The FAD was normal at all moments measured. CONCLUSIONS: Psychological morbidity of close relatives is significantly higher in the first months after the onset of GBS. The patient's condition has an important impact on the psychosocial functioning of close relatives. Therefore, a family approach is recommended to neurologist and other medical personnel during the first period of the disease. Also patient support groups may play a beneficial role for the relatives of GBS patients. PMID- 16476451 TI - Dynamic optimization over infinite-time horizon: web-building strategy in an orb weaving spider as a case study. AB - Dynamic state-dependent models have been widely developed since 1990s for solving questions in evolutionary ecology. Up to now, these models were mainly run over finite-time horizon. However, for many biological questions an infinite-time horizon perspective could be more appropriate, especially when the end of the modeled period is state- rather than time-dependent. Despite this approach is widely used in the field of economics and operational research, thus far no work has been providing biologists with a general method to solve infinite-time horizon problems. Here we present such a method, through the exhaustive description of an algorithm that we implement to determine the strategy an organism should follow to reach a particular state as fast as possible while limiting mortality risk. To illustrate that method we explored web-building behavior in an orb-weaving spider. How are adult females predicted to build their successive webs to gain energy, grow, and lay their first clutch as fast as possible, without suffering from either predation or starvation? From this example, we first show how an optimal strategy over infinite-time horizon can be processed and selected. Second, we analyse variations of the optimal web-building strategy along with the spider's body weight and predation risk during web building. Our model yields two main predictions: (1) spiders reduce their web size as they are gaining weight due to body-mass-dependent cost of web-building behavior, and (2) this reduction in web size starts at lower weight under higher predation risk. PMID- 16476452 TI - Exaggerated local hand sympathetic but not renin-angiotensin system activation in patients with primary Raynaud's phenomenon. AB - In Raynaud's disease (RD), an overactivity of sympathetic nervous system (SNS) was hypothesized but only indirect proofs were obtained. Complex interactions between the renin-angiotensin system (RAS) and SNS were reported without clear demonstrations of a RAS involvement. Recently, the use of ACE inhibitors and AT1 receptor antagonists in RD patients showed mixed results. The study of total and regional kinetics of tritiated noradrenaline (NA) and the measurements of local Angiotensin (Ang) I and II arterial-venous gradient were performed in 10 RD patients and 10 controls both in rest conditions and following a cold pressor test (CPT). Hand blood flow (HBF) was measured by strain-gauge plethysmography. Baseline HBF was slightly lower in patients than in controls, but during CPT, it significantly decreased only in RD patients (P < 0.01). Total (3H)-NA clearance and spillover were similar in the two groups throughout the study. On the contrary, baseline hand NA spillover was higher in RD patients than in controls and the difference further increased during CPT. Hand NA spillover was linearly related to HBF (P < 0.001). Arterial-venous Ang I and Ang II gradients were positive without difference between controls and patients throughout the study. In conclusion in RD patients, a pathological waste of NA from sympathetic nervous endings of the hand region, exaggerated by sympathetic stimulation, occurs but an enhanced local Ang II formation was not demonstrable. PMID- 16476453 TI - A health promotion programme in Adventist and non-Adventist women based on Pender's model: a pilot study. AB - OBJECTIVE: In the last decades, the epidemiological and demographic transition in Chile has resulted in a considerable increase in diet-related non-communicable chronic diseases. The aim of this study was to determine the impact of a health promotion programme (HPP) on behaviour in terms of the dimensions of the health promoting lifestyle profile (HPLP) based on Pender's model. Pender based his research on the development of a model that explains, predicts and modifies forms of behaviour that promote health; an HPP based on this model may achieve positive changes that promote a healthy lifestyle. STUDY DESIGN: This was a comparative study and the participants were Seventh-Day Adventist Church women (SDAW) and non Seventh-Day Adventist Church women (NSDAW) aged 20-45 years from the urban area of Villarrica, Ninth Region of Chile. From a population of 300 women (150 SDAW and 150 NSDAW), a random sample of 18 SDAW and 18 NSDAW was chosen. Both groups were comparable in age and socio-economic status. An HPP was undertaken for both groups between April and September 2002. The response rate was 100%. METHODS: The overall score and the scores for the six dimensions of the HPLP (self actualization, health responsibility, exercise, nutrition, interpersonal support and stress management) were measured in the pre- and post-test periods. Statistical analysis was performed using the Sign test and Wilcoxon's test. Data were processed using the statistical analysis system. RESULTS: In both groups, the median scores increased significantly between the pre- and post-test periods for the overall HPLP score and the scores of the six dimensions. When comparing the median scores in SDAW with NSDAW, only the score for nutrition was significantly higher in SDAW than NSDAW in both the pre-test (P<0.0001) and post test (P<0.0005) periods. CONCLUSIONS: These findings demonstrate that an HPP based on Pender's model improves behaviour related to a health-promoting lifestyle in both groups, but more so for the NASDW. PMID- 16476454 TI - Patients' awareness of their rights in a developing country. AB - AIM: To determine the awareness of a regulation on patients' rights passed in 1998 in Turkey. METHODS: All hospitalized patients in the internal medicine and general surgery wards in three large hospitals during the study period were eligible for the study, which was performed in 2001 in the Denizli province, Turkey. The study group consisted of 166 patients. Data were collected during face-to-face interviews using a questionnaire. RESULTS: Only 9% of patients were aware of the regulation about patients' rights. Most patients were given equal access to health care (91%), benefitted from the capabilities of their selected healthcare institutions (86.7%), and their privacy was protected (86.1%). Those patients who stated that they had not received care in accordance with their rights identified the shortcomings as not receiving written (95.2%) or verbal (53%) information from healthcare professionals, and the failure of healthcare personnel to introduce themselves (75.3%). On most occasions, the patients stated that they were not able to request services as stated in the PRR from healthcare professionals. CONCLUSION: Few patients knew about the regulation on patients' rights, indicating a need for extensive education of patients and healthcare professionals. PMID- 16476455 TI - An overview of illegal opioid use and health services utilization in Canada. AB - OBJECTIVES: Systematic research on health and treatment services availability for and utilization by illegal opioid users in Canada are very limited. Comparative data across provinces and territories is almost entirely absent. This study was designed to provide an overview of illegal opioid use and health services utilization among illegal opioid users across Canada. METHODS: A combination of statistical data and key informant data was used. Surveys were sent to key informants in all provinces and territories of Canada. Survey questions covered the number of illegal opioid users in each province, the number of opioid users receiving methadone maintenance treatment (MMT), the number of physicians authorized to prescribe methadone, and the number of opioid users receiving other outpatient and inpatient treatment. In addition, relevant data were collected from several statistical sources, both provincial and federal. The number of substance-use-related overdose deaths was obtained from the provincial coroners' offices. RESULTS: It is estimated that there were more than 80,000 regular illegal opioid users in Canada in 2003. The most prevalent treatment utilized was MMT; about one-quarter (26%) of the estimated opioid users received this type of treatment in 2003. Other forms of outpatient and inpatient treatment were of only minor importance compared with MMT. The number of illegal drug-related overdose deaths in Canada was 958 in 2002. Rates of drug use, health services utilization and overdose deaths showed considerable variation by province. CONCLUSIONS: Although the opioid use treatment system in Canada has expanded in recent years, especially with respect to the availability of MMT, the treatment utilization rates are still lower than in most countries in Western Europe. Rates of current treatment utilization as well as the relatively high number of overdose deaths suggest that there is still room for improvement in the Canadian health and social care system with respect to opioid use. PMID- 16476456 TI - A comparative study of behavioural, physical and mental health status between term-limited and tenure-tracking employees in a population of Japanese male researchers. AB - OBJECTIVES: Traditional lifelong employment systems have been changing rapidly in Japan. The aim of this study was to assess the health impacts of term-limited employment systems that have recently been introduced into Japanese academic institutes. STUDY DESIGN: Cross-sectional. METHODS: A total of 514 male researchers (275 term limited and 239 tenure track) were compared in terms of behavioural, physical and mental status at annual health examinations. At these examinations, working hours and health-related lifestyles were examined using a self-completed questionnaire. Clinical structured interviews of the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) were used to detect major depression. RESULTS: The term-limited researchers tended to work longer hours (P<0.001), drink alcohol more frequently (P<0.001) and eat breakfast less regularly (P<0.001) compared with the tenure-track researchers. Although the prevalence of DSM-IV major depression was comparable (P>0.05) between the two groups, fatigue was more prevalent (P=0.027) in the term-limited researchers than in the tenure-track researchers, adjusting for the effects of age. Compared with colleagues working in the same laboratories, the term-limited researchers worked longer hours, irrespective of fatigue, whereas only the fatigued tenure-track researchers worked longer hours. In the total sample, the fatigued researchers tended to belong to laboratories where their colleagues, on average, worked longer hours, compared with the non-fatigued researchers. CONCLUSIONS: These results imply that the term-limited researchers suffered more from fatigue, due to longer working hours, than their colleagues, and that organized, rather than personal, interventions with respect to the working environment may be effective in reducing overload in such workplaces. PMID- 16476457 TI - Inhibition of estrone sulfatase by aromatase inhibitor-based estrogen 3 sulfamates. AB - Our rationale is based on the finding that estrone 3-sulfamate (EMATE, 2d), a typical estrone sulfatase (ES) inhibitor, can be hydrolyzed and the pharmacological effect of the free estrogen contributes to the bioactivity of the sulfamate. A number of 3-sulfamoylated derivatives of the good aromatase inhibitors, 2- and 4-halogeno (F, Cl, and Br) estrones and their estradiol analogs as well as 6beta-methyl and phenyl estrones, were synthesized and evaluated as inhibitors of ES in human placental microsomes in comparison with the lead compound EMATE. Among them, 2-chloro- and 2-bromoestrone 3-sulfamates (2b and 2c), along with their estradiol analogs 3b and 3c, were powerful competitive inhibitors with K(i)'s ranging between 4.0 and 11.3 nM (K(i) for EMATE, 73 nM). These four sulfamates as well as the 2-fluoro analogs 2a and 3a inactivated ES in a time-dependent manner more efficiently than EMATE, and 2 halogeno estrone sulfamates 2 also caused a concentration-dependent loss of ES activity. The results may be useful for developing a new class of drugs having a dual function, ES inhibition and aromatase inhibition, for the treatment of breast cancer. PMID- 16476458 TI - Role of the sodium-dependent phosphate co-transporters and of the phosphate complexes of uranyl in the cytotoxicity of uranium in LLC-PK1 cells. AB - Although uranium is a well-characterized nephrotoxic agent, very little is known at the cellular and molecular level about the mechanisms underlying the uptake and toxicity of this element in proximal tubule cells. The aim of this study was thus to characterize the species of uranium that are responsible for its cytotoxicity and define the mechanism which is involved in the uptake of the cytotoxic fraction of uranium using two cell lines derived from kidney proximal (LLC-PK(1)) and distal (MDCK) tubule as in vitro models. Treatment of LLC-PK(1) cells with colchicine, cytochalasin D, concanavalin A and PMA increased the sodium-dependent phosphate co-transport and the cytotoxicity of uranium. On the contrary, replacement of the extra-cellular sodium with N-methyl-D-glucamine highly reduced the transport of phosphate and the cytotoxic effect of uranium. Uranium cytotoxicity was also dependent upon the extra-cellular concentration of phosphate and decreased in a concentration-dependent manner by 0.1-10 mM phosphonoformic acid, a competitive inhibitor of phosphate uptake. Consistent with these observations, over-expression of the rat proximal tubule sodium dependent phosphate co-transporter NaPi-IIa in stably transfected MDCK cells significantly increased the cytotoxicity of uranium, and computer modeling of uranium speciation showed that uranium cytotoxicity was directly dependent on the presence of the phosphate complexes of uranyl UO(2)(PO(4))(-) and UO(2)(HPO(4))(aq). Taken together, these data suggest that the cytotoxic fraction of uranium is a phosphate complex of uranyl whose uptake is mediated by a sodium dependent phosphate co-transporter system. PMID- 16476459 TI - DNA damage, redox changes, and associated stress-inducible signaling events underlying the apoptosis and cytotoxicity in murine alveolar macrophage cell line MH-S by methanol-extracted Stachybotrys chartarum toxins. AB - Spore-extracted toxins of the indoor mold Stachybotrys chartarum (SC) caused cytotoxicity (release of lactate dehydrogenase), inhibition of cell proliferation, and cell death in murine alveolar macrophage cell line MH-S in a dose- and time-dependent manner. Apoptotic cell death, confirmed based on morphological changes, DNA ladder formation, and caspase 3/7 activation, was detectable as early as at 3 h during treatment with a toxin concentration of 1 spore equivalent/macrophage and was preceded by DNA damage beginning at 15 min, as evidenced by DNA comet formation in single cell gel electrophoresis assay. The apoptotic dose of SC toxins did not induce detectable nitric oxide and pro inflammatory cytokines (IL-1beta, IL-6, and TNF-alpha) but showed exacerbated cytotoxicity in presence of a non-apoptotic dose of the known pro-inflammatory agent LPS (10 ng/ml). Intracellular reduced glutathione (GSH) level showed a significant decrease beginning at 9 h of the toxin treatment whereas oxidized glutathione (GSSG) showed a corresponding significant increase, indicating a delayed onset of oxidative stress in the apoptosis process. The toxin-treated macrophages accumulated p53, an indicator of DNA damage response, and showed activation of the stress-inducible MAP kinases, JNK, and p38, in a time-dependent manner. Chemical blocking of either p38 or p53 inhibited in part the SC toxin induced apoptosis whereas blocking of JNK did not show any such effect. This study constitutes the first report on induction of DNA damage and associated p53 activation by SC toxins, and demonstrates the involvement of p38- and p53 mediated signaling events in SC toxin-induced apoptosis of alveolar macrophages. PMID- 16476460 TI - Analysis of a baculovirus lacking a functional viral fibroblast growth factor homolog. AB - Baculoviruses encode fibroblast growth factor (vfgf) homologs whose function during virus infection is unknown. We constructed a recombinant bacmid of Autographa californica M nucleopolyhedrovirus (AcMNPV) lacking a functional vfgf and characterized it in two insect cell lines. The kinetics of budded virus production were similar in the parental and vfgf-deficient viruses in both cell lines at both high and low multiplicities of infection. In addition, no obvious differences were observed between the mutant and parental viruses in protein or DNA synthesis. Finally, coinfection of vfgf-containing and -deficient viruses and passage for several generations did not reveal a consistent growth advantage for either virus. PMID- 16476461 TI - Rescue of the prototypic Arenavirus LCMV entirely from plasmid. AB - We document a helper-independent reverse genetics system for rescuing infectious arenaviruses from cloned cDNAs. We constructed plasmids containing full-length cDNAs of the antigenomic (ag) L and S segments of the Armstrong (ARM) strain of the prototypic Arenavirus lymphocytic choriomeningitis virus (LCMV) flanked at their 5'- and 3'-termini by the T7 RNA polymerase (T7RP) promoter and ribozyme sequences, respectively. These plasmids directed intracellular synthesis of viral L and S ag RNA species in cells expressing plasmid-supplied T7RP. Co-expression of plasmid-supplied LCMV trans-acting factors, nucleoprotein (NP) and polymerase (L), resulted in replication and expression of L and S ag and genome RNA species, and generation of LCMV infectious progeny termed rT7/LCMV. The recombinant rT7/LCMV was unequivocally identified based on a genetic tag introduced in the recombinant S segment. In addition, rT7/LCMV exhibited growth and biological properties predicted for an ARM-like LCMV. To our knowledge, this is the first documented Arenavirus rescue, as well as of an ambisense negative strand (NS) RNA virus, entirely from cloned cDNAs. Our results extend the use of reverse genetic approaches for DNA-mediated virus rescue to all known virus families with NS RNA genome. PMID- 16476462 TI - Human cytomegalovirus infection modulates DNA base excision repair in fibroblast cells. AB - Regulation of DNA repair mechanisms during the viral replication cycle may have consequences for the virus with regards to genomic variability, adaptation, and replication of viral DNA. We have studied the activities and expression patterns of key enzymes involved in the first two steps of base excision repair (BER) of DNA in primary fibroblasts infected by human cytomegalovirus (HCMV). Infected cells were very proficient for removal of uracil and 5' hydrolysis of AP sites (AP endonuclease activity) as compared to the mock-infected cells, suggesting a direct role in generating free ends at uracil lesions in DNA for initiation of viral replication. Furthermore, the capacity to initiate repair of alkylated and oxidized base lesions were reduced in HCMV-infected cells, indicating increased mutation frequencies that could promote genetic variability. We hypothesize that modulation of BER activities may play an important role in HCMV pathogenesis to ensure efficient replication and genomic variation of viral DNA. PMID- 16476463 TI - Spatial and feature-based effects of exogenous cueing on visual motion processing. AB - In two experiments, we investigated the effects of exogenous cueing on visual motion processing. The first experiment shows that the typical pattern of reaction time (RT) effects, namely early facilitation and later inhibition of return (IOR), can be obtained using a color change as exogenous cue and a direction change as target. In the second experiment, we manipulated the validity of the cue independently with respect to location and feature using transparent motion stimuli. Facilitation of RTs with short cue-target interstimulus-intervals (ISIs) was only evident for targets with both the valid location and the valid feature. Furthermore, at longer cue-target intervals, RTs were prolonged for targets at the cued location, irrespective of the cued feature. These results demonstrate spatial and feature-based components of early facilitation and purely spatial IOR. PMID- 16476464 TI - Only two phase mechanisms, +/-cosine, in human vision. AB - We evaluated the proposal that there exist detectors of the following four cardinal phases in human vision: +cosine, -cosine, +sine, and -sine. First, we assessed whether there was evidence that these cardinal phases were processed by independent 'labeled lines,' using a discrimination at detection threshold paradigm. Second, we assessed whether suprathreshold phase discrimination was best at phases intermediate between these cardinal values. Third, we tried to replicate previous evidence showing that an absence of facilitation occurs only between cosine pedestals and sine tests (or vice-versa). In all three experimental approaches we found no compelling evidence for four cardinal phase groupings. We did however find evidence for independent detectors for pure increments and decrements (+/-cosine). We suggest that phase discrimination, whether at threshold or suprathreshold, is mediated by mechanisms that encode the relative positions and contrasts of local increments and decrements within the stimulus. PMID- 16476465 TI - Comparative study of reactions of endocrine disruptors bisphenol A and diethylstilbestrol in electrochemical treatment and chlorination. AB - Transformations of diethylstilbestrol (DES) and bisphenol A (BPA) in conventional chlorination and electrochemically (EC) treated solutions were examined using spectrophotometry and chromatographic analyses. EC treatment was carried out using an undivided EC cell with a PbO2 anode and a stainless steel cathode. EC treatment and conventional chlorination caused DES and BPA to undergo a rapid degradation accompanied by the generation of low molecular weight chlorinated organic species indicative of the breakdown of DES and BPA. The identified compounds were predominated by chloroacetic acids (HAAs), but approximately 80% of the total organic halogen (TOX) was comprised by unidentified species. For EC treatment, the HAA yields were lower and HAAs were predominated by monochloroacetic acid (MCAA), while in the case of conventional chlorination, trichloroacetic acid (TCAA) was predominant and MCAA was virtually absent. The changes in the HAA speciation and yields were concluded to be caused by the EC driven reductive dehalogenation which, however, did not affect the unidentified fraction of TOX. This indicated that the unidentified part of TOX was comprised by aromatic chlorinated forms of BPA and DES. Their resistance to degradation in EC reactors indicates that these compounds may be stable in conditions typical for drinking water treatment and distribution. PMID- 16476466 TI - New measurement principle and basic performance of high-sensitivity turbidimeter with two optical systems in series. AB - An outbreak of Cryptosporidium infection from tap water has triggered the implementation of stricter controls for the turbidity of filtered water in Japan. Accordingly, a growing number of water purification plants have begun to measure the number of particles, which is more closely correlated to this kind of protozoan than turbidity. With this background we have developed a high sensitivity turbidimeter based on the particle counting method that supports both turbidity measurement and particle counting to realize a compact and low-cost monitoring system of filtered water. This paper explains the basic principles of the turbidimeter, which uses both forward-angle light scattering and light obscuration methods in series and determines the turbidity from particle counting by applying Mie's theory of light scattering. In basic experiments on turbidity standards such as kaolin solution and formazine solution, we verified that the turbidimeter could determine the particle counts and turbidity of filtered water with sufficient accuracy. PMID- 16476467 TI - Effect of 2,4-D contamination on soil functional stability evaluated using the relative soil stability index (RSSI). AB - Soil functional stability is the capacity of soil functions to resist and recover from an environmental perturbation and can be used to evaluate soil health. It can be influenced by the presence of xenobiotics such as herbicides. The impact of a fresh 2,4-D contamination (36 mg kg(-1) dry soil) on soil functional stability was evaluated by comparing the capacity of soil enzyme activities to resist and recover from a heat perturbation for both a clean and 2,4-D contaminated soil. The functional stabilities of the soils (uniform sands, pH 6.9, 7% (w/w) organic matter) were calculated using the relative soil stability index (RSSI). The RSSI scores indicate the proportion of potential enzyme activity the soil retains after a perturbation compared to the potential activity of an unperturbed soil. Six extra-cellular enzyme activities (acid and alkaline phosphatases, arylsulfatase, urease, protease and beta-glucosidase) were monitored in soil microcosms during a 15-day period. During this period, a 60 degrees C heat perturbation was applied to the soil for 24 h. The activities of arylsulfatase and protease were found to be the most stable following heat perturbation obtaining the highest RSSI scores (87% and 77%, respectively). Urease activity showed the lowest RSSI score (38%). Although all enzyme activities were inhibited by the presence of 2,4-D, the RSSI results indicated that contamination lowered the stability of only three enzyme activities (arylsulfatase, beta-glucosidase and urease). The RSSI adequately described resistance, recovery and recovery rate parameters and enabled differentiation between functional stabilities of clean and contaminated soil and between different soil types. PMID- 16476468 TI - Role of surface functionality in the adsorption of anionic dyes on modified polymeric sorbents. AB - While synthetic polymeric sorbents effectively treat dye wastewaters by adsorption, the underlying mechanisms remain to be understood. This work determined the adsorption of an anionic dye by three polymers differing significantly in surface functionality. Surface functional groups of polymers were indicated in FTIR spectra and quantified by the Boehm titration. In reference to the commercial sorbent XAD-4 with a low degree of functionality, the laboratory synthesized NG-8 had primarily acidic functional groups, whereas its aminated product MN-8 had mainly basic amino groups. Electrophoresis determined the points of zero charge of 4.18, 3.23, and 4.51 for XAD-4, NG-8, and MN-8, respectively. The adsorption of Reactive Black 5 dye (RB5) by all the sorbents on a unit surface area basis increased with decreasing pH. At the same low pH (4.40), the adsorption by NG-8 was similar to that by XAD-4, indicating little influence of protonated (neutral) surface functional groups. In contrast, the adsorption by NG-8 at pH 6.05 was 75% lower than that by XAD-4, resulting apparently from the strong electrostatic repulsion between RB5 and deprotonated (negative) groups. Amination substantially enhanced RB5 adsorption by eliminating acidic groups and creating a positive charge on the surface of MN-8. The adsorptive enhancement was also achieved in the presence of CaCl2, due presumably to the neutralization of negative surface charge by Ca2+ and the RB5-Ca2+ pairing. These results manifest the important role of surface functionality in the adsorption of dyes by synthetic polymers. PMID- 16476469 TI - Efficacy of anti-angiogenic treatment of tumors in old versus young mice. AB - Cancer treatment in the older population, the most afflicted by the disease, is as yet, inefficient. A reduced aggressiveness of tumors is often observed in the elderly, implying the necessity for therapeutic modalities adjusted to age. A rational design of age-related cancer therapy could be based on the mechanisms of this phenomenon. It is suggested that, in addition to the patient's old age specific health problems (which prohibit the use of the aggressive cancer treatments now in use), the age-related differential tumor biology (apparently beneficial to the old) should also be considered for the design of treatment modalities suitable for the aged. Based on one mechanism of the reduced aggressiveness of tumors in the old (age-dependent decreased angiogenesis), we compared the effect of an anti-angiogenic treatment in young and old mice. TNP 470 treatment resulted in an inhibitory effect on B16 melanoma in both young and old mice but the effect was more pronounced in old animals. Moreover, a high percentage of long-term surviving animals was observed only in the old-treated mice. Treatment with TNP-470 of the AKR lymphoma produced similar results. We thus found a differential age-dependent therapeutic efficiency of an anti angiogenic agent on two tumors. Importantly, the anti-angiogenic drug was more efficient against tumors of old animals. PMID- 16476470 TI - The levels of circulating markers of atherosclerosis and inflammation in subjects with different degrees of body mass index: Soluble CD40 ligand and high sensitivity C-reactive protein. AB - BACKGROUND: It is well demonstrated that obesity is an independent risk factor for cardiovascular diseases. Recent studies have shown that obesity, insulin resistance and atherosclerosis are closely related phenomena in which low-grade inflammatory state and prothrombotic condition has pivotal roles. It has been shown that CD40-soluble CD40 ligand (sCD40L) interactions might constitute an important mediator for vascular inflammation. The aim of the present study was to assess sCD40L in relation to hs-CRP and cardiovascular risk factors in relation to body mass index (BMI). MATERIALS AND METHODS: Serum sCD40L and hs-CRP concentrations were measured in 52 obese patients and 28 non-obese subjects by ELISA. Insulin resistance was calculated by homeostasis model assessment-insulin resistance (HOMA-IR). We divided the participants into three groups depending in their BMI levels (Group 1: BMI <25 kg/m(2), Group 2: BMI 30-34.9 kg/m(2), Group 3: BMI > or =35 kg/m(2)). RESULTS: We determined that the mean sCD40L of group 3 was significantly higher than group 1 and group 2 (p<0.05, p<0.05, respectively). However, there was no significant correlation between plasma sCD40L levels and BMI. Plasma levels of hs-CRP were higher in obese group than the non-obese group (p<0.001). The levels of sCD40L were not significantly different between the two groups. The mean hs-CRP levels increased gradually in accordance with groups of BMI, there was a strong correlation between hs-CRP levels and BMI (r=0.724, p<0.001). There was no significant correlation between sCD40L and hs-CRP levels in all participants. CONCLUSIONS: It is still a subject for debate whether sCD40L levels are increased or not in obesity. However, the results of this study showed that sCD40L is substantially increased in patients with severe obesity. In terms of causality, the relatively small sample size and cross-sectional design of this study are considered to be the limitation factors. PMID- 16476472 TI - Palliative chemotherapy "near the end-of-life" in ovarian cancer: not necessarily "aggressive cure-oriented therapy". PMID- 16476471 TI - Single-dose actinomycin D: Efficacy in the prophylaxis of postmolar gestational trophoblastic neoplasia in adolescents with high-risk hydatidiform mole. AB - OBJECTIVE: To evaluate the efficacy of a single prophylactic dose of actinomycin D (Act-D) in the reduction of postmolar gestational trophoblastic neoplasia (GTN) in adolescents with high-risk hydatidiform mole (Hr-HM). METHODS: In a retrospective study, 60 adolescents with Hr-HM were selected from a cohort of patients with gestational trophoblastic disease (GTD) followed at Santa Casa, Porto Alegre, Brasil. Twenty-nine received a single dose of Act-D at the time of uterine evacuation as prophylactic chemotherapy (P-chem) (study group) and 31 patients with the same risk factors did not received P-chem (control group). Patient follow-up was the same in both groups. Each group was analyzed for number of adolescents with postmolar GTN, morbidity associated with postmolar GTN, and reproductive outcomes. RESULTS: Postmolar GTN was diagnosed in two (6.9%) adolescents (95% CI, 0.0-16.1) in the study group and in 9 (29.0%) patients (95% CI, 13-45) in the control group. The reduction of postmolar GTN with a single dose of Act-D used as P-chem was 76% (relative risk = 0.24; 95% CI, 0.06-0.99). Adverse effects of P-chem were minor. In the follow-up, when postmolar GTN were diagnosed, severity of disease was not increased, compliance with follow-up was not reduced, and reproductive outcomes after discharge were similar. CONCLUSIONS: P-chem with a single dose of Act-D reduced postmolar GTN in 76% during follow-up of adolescents with Hr-HM. Since this regimen may reduce treatment costs, without affecting compliance with follow-up, it can be adopted by any Trophoblastic Disease Center. PMID- 16476474 TI - Occult endometrial cancer and decision making for prophylactic hysterectomy in hereditary nonpolyposis colorectal cancer patients. AB - BACKGROUND AND OBJECTIVE: Hereditary nonpolyposis colorectal cancer (HNPCC) is the most frequent form of hereditary colorectal cancer. In addition to the high lifetime risk for colorectal cancer in mutation carriers, there is also a remarkably increased risk for endometrial cancer (EC). METHODS: In this retrospective study, clinical and molecular approach to the individual decision making as to whether or not to perform a prophylactic hysterectomy in a subset of HNPCC patients is discussed. 147 female patients meeting at least one criterion of the Bethesda guidelines were included in this analysis between 1995 and 2003. After clinical and genetic counseling, patients gave informed written consent and microsatellite analysis, immunohistochemistry and sequencing of the mismatch repair genes MLH1, MSH2 and MSH6 was performed. RESULTS: 11 of the analyzed patients had a personal history of EC and had undergone previous hysterectomy at ages 26 to 62 years. Prophylactic hysterectomy with oophorectomy was considered in postmenopausal women meeting the Amsterdam criteria and/or carrying a disease causing mismatch repair gene mutation who were operated on because of diagnosed colorectal cancer in our center for hereditary cancer. This procedure was performed in 4 patients. None of them had shown any symptoms of a gynecologic malignancy. Preoperative gynecological examination showed no evidence for EC or ovarian cancer in these patients. Postoperative histological examination showed EC stage T1b N0 M0 in 2 patients. CONCLUSIONS: Since the efficiency of gynecological surveillance is uncertain, prophylactic hysterectomy could be an option for a subset of HNPCC patients and mutation carriers. PMID- 16476475 TI - Obesity and leisure time physical activity among Canadians. AB - BACKGROUND: The association between physical activity and obesity are age and sex related. METHODS: The analysis was based on data from 81,512 Canadians aged 20 to 64 years who participated in the Canadian Community Health Survey conducted in 2000-01. Information collected included body mass index (BMI:<25.0 kg/m(2) (normal weight); 25.0-29.9 kg/m(2) (overweight); and 30.0+ kg/m(2) (obese)) and energy expenditure (EE) calculated based on the frequency and duration of leisure time physical activity (LTPA) and its value of metabolic energy cost. RESULTS: Obese men and both obese and overweight women had lower EE values compared with those with normal weight. Overweight men 30 to 49 years of age were more active than men with normal weight. Average EE values decreased with age. Overall, 53.2% of men and 57.1% of women were inactive (EE<1.5 kcal/kg/day). In obese men and women, the corresponding prevalences were 59.7% and 71.4%, respectively. The prevalence of physical inactivity increased with increasing BMI in women and there was no "dose-effect" relationship in men. CONCLUSIONS: Obese Canadians are less active, but overweight men tend to be more active than those with normal weight. The variation in the associations across age and sex groups suggests that intervention program development should take age and sex into consideration. PMID- 16476476 TI - Use of enrofloxacin in the treatment of canine brucellosis in a dog kennel (clinical trial). AB - To date, no totally effective antibiotic for the eradication of canine brucellosis has been found. The purpose of this study was to evaluate the efficacy of enrofloxacin in a kennel infected with Brucella canis. Twelve dogs, 2 males and 10 females (including 1 in estrus, 3 pregnant, and 6 in anestrus) infected with B. canis were given 5 mg/kg of enrofloxacin orally every 12 h for 30 days. Females received additional courses of enrofloxacin during the estral and luteal phases of the subsequent cycles (0-2 cycles). They were repeatedly mated by infected males. A serological follow-up was carried out for 38 months. The clinical, serological and bacteriological findings were recorded. In a trial carried out 14 months after the beginning of this study, all dogs were negative on the Rapid Slide Agglutination Test (RSAT). No abortions were observed. All mated female dogs conceived and gave birth to healthy puppies. Cultures of postpartum vaginal discharges (lochia) were negative for B. canis. Similar to other treatments, although enrofloxacin was not completely efficacious in treating canine brucellosis, it maintained fertility and avoided the recurrence of abortions, transmission of the disease to the puppies and dissemination of microorganisms during parturition. We inferred that enrofloxacin could be used as an alternative drug for the treatment of canine brucellosis. PMID- 16476477 TI - Dynamic mechanical studies of hydrolytic degradation in isotropic and oriented Maxon B. AB - Hydrolytic degradation studies have been undertaken on Maxon B, a bioresorbable block copolymer of polyglycolic acid (PGA) and polytrimethylene carbonate (TMC). Isotropic and oriented samples were studied by dynamic mechanical measurements over a wide range of temperatures. In addition to mechanical tests, water content and mass loss were also determined on the degraded samples. At early stages of degradation water content was the dominant factor and plasticisation lead to reductions in the glass transition temperatures of the PGA and TMC components. Orientation was shown to give significant improvements in the mechanical properties, including overall increases in modulus and an increase in the glass transition temperature of the PGA component, which is important for the behaviour at body temperature (37 degrees C). Oriented samples also showed significantly less reduction in mechanical properties on degradation. Simple one-dimensional Takayanagi models were used to provide useful insight into the understanding of the mechanical behaviour. PMID- 16476479 TI - Pattern of immunoglobulin and T-cell receptor (Ig/TCR) gene rearrangements in Polish pediatric acute lymphoblastic leukemia patients--implications for RQ-PCR based assessment of minimal residual disease. AB - We studied 23 Polish children with precursor-B-ALL, using PCR-heteroduplex analysis and DNA sequencing, to determine the availability of Ig/TCR gene rearrangements as patient-specific MRD-RQ-PCR targets. We found IGH, IGK-Kde, incomplete TCRD, Vdelta2-Jalpha, TCRG and TCRB rearrangements in 83%, 39%, 61%, 35%, 61% and 13% of patients, respectively. Comparison of Ig/TCR gene rearrangements pattern (frequency and characteristics of rearrangements) in Polish patients with those reported for patients of other European nationalities did not show major differences. These results are the first promising step for further development of MRD study in Polish patients according to current diagnostic standards. PMID- 16476478 TI - The effect of nanotopography on calcium and phosphorus deposition on metallic materials in vitro. AB - To date, long-term functions of osteoblasts leading to calcium and phosphorus mineral deposition on nanometals have not been determined. Nanometals are metals with constituent metal particles and/or surface features less than 100 nm in at least one dimension. For this reason, the objective of this in vitro study was to determine the amount of calcium and phosphorus mineral formation on microphase compared to nanophase Ti, Ti6Al4V, and CoCrMo cultured with and without osteoblasts (bone-forming cells). The results of this study provided the first evidence of significantly greater calcium and phosphorus deposition by osteoblasts and precipitation from culture media without osteoblasts on nanophase compared to respective microphase Ti6Al4V and CoCrMo after 21 days; the greatest calcium and phosphorus mineral deposition occurred on nanophase CoCrMo while the greatest calcium and phosphorus mineral precipitation without osteoblasts occurred on nanophase Ti6Al4V. No differences were found for any type of Ti: wrought, microphase, or nanophase. Moreover, increased calcium and phosphorus mineral content correlated to greater amounts of underlying aluminum content on Ti6Al4V surfaces. Since, compared to microphase Ti6Al4V, nanophase Ti6Al4V contained a higher amount of aluminum at the surface (due to greater surface area), this may provide a reason for enhanced calcium and phosphorus mineral content on nanophase Ti6Al4V. Regardless of the mechanism, this study continues to support the further investigation of nanometals for improved orthopedic applications. PMID- 16476480 TI - HSP70 gene expression in Mytilus galloprovincialis hemocytes is triggered by moderate heat shock and Vibrio anguillarum, but not by V. splendidus or Micrococcus lysodeikticus. AB - Complete sequence of HSP70 cDNA from the mussel, Mytilus galloprovincialis was established before quantifying its expression following moderate heat shock or injection of heat-killed bacteria. HSP70 cDNA is comprised of 2378 bp including one ORF of 654 aa, with a predicted 70 bp 5'-UTR and a 343 bp 3'-UTR (GenBank, 18 Jan 05, AY861684). Alignment identity ranged from 89% for Crassostrea ariakensis to 72% for C. virginica. Curiously, HSP70 gene and cDNA sequences from M. galloprovincialis, deposited later (03 and 27 May), show only 73% identity with the present sequence. Meanwhile, characteristic motifs of the HSP70 family were located in conserved positions. Expression of HSP70 gene was quantified on circulating hemocyte mRNA using Q-PCR after RT using random hexaprimers. Housekeeping gene was 28S rRNA. Four stresses were applied: heat shock that consisted of immersing mussels for 90 min at 30 degrees C and returning them to 20 degrees C sea water, one injection of heat-killed Gram-negative bacteria, Vibrio splendidus LGP32, one injection of heat-killed Gram-negative bacteria Vibrio anguillarum, one injection of heat-killed Gram-positive bacteria Micrococcus lysodeikticus. We found no significant modification of 28S rRNA gene expression. Significant increase of 5.2 +/- 0.4 fold the ratio HSP70/28S rRNA was observed 6 h after heat shock and was maximum at 15 h (6.1 +/- 1.1), and still significant after 24 h (1.7 +/- 0.03). Similarly, injecting V. anguillarum resulted in a significant increase of 2.7 +/- 0.1 after 12 h. Expression was maximum after 48 h (5.2 +/- 0.05) and returned to baseline after 72 h. In contrast, injecting V. splendidus or M. lysodeikticus failed to significantly modulate HSP70 gene expression at least during the first 3 days post-injection. Consequently, mussel hemocytes appeared to discriminate between pathogenic and non-pathogenic Vibrios, as well as between Gram-negative and Gram-positive bacteria. PMID- 16476482 TI - Staphylococcus aureus bacteraemia in children and neonates: a 10 year retrospective review. AB - Rates of Staphylococcus aureus bacteraemia (SAB) are published performance indicators for hospital-acquired infection. In adults SAB is often associated with central venous catheters (CVC), mortality is high and up to 40% are MRSA. However, there is little data on SAB in neonates and children in the UK. AIM: To describe the presentation, management and outcome of SAB on a neonatal and paediatric unit in a District General Hospital (DGH) over a 10 year period. METHOD: Case notes of children<16 years with SAB between May 1993 and April 2003 were studied. SAB which developed >48 h after admission was defined as hospital acquired. Contamination was probable if the clinical picture was unsupportive of infection, or if repeat culture was negative and no treatment was given. RESULTS: Neonatal unit: Thirty-three of 40 episodes were reviewed (median gestation 32 weeks, median age 21 days). Three of 33 (9%) were contaminants. All SAB were hospital acquired. Twenty-six of 30 (87%) had non-specific presentation, but 15 developed a focus of infection (skin 12, chest 3). Seventeen (57%) infants had CVCs. Eight (27%) infants had MRSA bacteraemia, seven with CVCs. Three (10%) infants died. Paediatric unit: Sixty-four of 70 episodes were reviewed (median age 2 years). Thirteen of 64 (20%) were contaminants. Ten of 51 (20%) were hospital acquired. Presentations were with skin infection 18, bone/joint infection 13, non-specific 13, respiratory 8. Only two had MRSA, one with CVC. One (2%) child died, from an unrelated cause. CONCLUSION: SAB on a paediatric unit shows a very different pattern compared to SAB in adults. The pattern on a neonatal unit is more similar to that in adults. Both children and neonates have a lower mortality and a lower incidence of MRSA, whilst paediatric SAB has a weaker association with CVC. The proportion of SAB which is hospital acquired is low on a paediatric unit, making SAB an unreliable performance indicator. Most SA in blood cultures are not due to contamination. Prospective studies are needed to determine appropriate investigation and treatment. PMID- 16476481 TI - Decreased expression of inflammation-related genes following inhalation exposure to manganese. AB - Excessive exposure to manganese (Mn) by inhalation can induce psychosis and Parkinsonism. The clinical manifestations of Mn neurotoxicity have been related to numerous physiological and cellular processes, most notably dopamine depletion. However, few studies have explored the molecular events that are triggered in response to exposure to Mn by inhalation. In this current study, the transcriptional patterns of genes related to oxidative stress or inflammation were examined in the brain rats of exposed to inhaled Mn during either gestation or early adulthood. The expression of genes encoding for proteins critical to an inflammatory response and/or possessing pro-oxidant properties, including TGFbeta and nNOS, were slightly depressed by prenatal exposure, whereas inhalation exposure to Mn during adulthood markedly down-regulated their transcription. However, when exposures to manganese occurred during gestation, the extent of altered gene expression induced by subsequent exposure to Mn in adulthood was reduced. This suggests that prior exposure to Mn may have attenuated the effects of inhalation exposure to Mn in adulthood, in which the expression of inflammation-related genes were suppressed. PMID- 16476483 TI - Prevalence of Haarlem I and Beijing types of Mycobacterium tuberculosis strains in Iranian and Afghan MDR-TB patients. AB - OBJECTIVES: This survey identified the spoligopatterns of Mycobacterium tuberculosis strains with an international designation responsible for transmission and prevalence of Multi-Drug Resistance Tuberculosis (MDR-TB) among native and immigrant population of Tehran (2000-2005). METHODS: The spacer oligonucleotides typing was performed on 263 M. tuberculosis strains isolated from verified cases of MDR-TB. Clinical and demographical data of patients were collected using traditional methods. RESULTS: Classical epidemiological investigation revealed that out of 263 MDR-TB cases, 175, 66.5% were isolated from Afghan immigrants. In both communities, majority of MDR-TB cases had either previous history of TB (107, 40.6%) or had a close contact (84, 31.9%). By spoligotyping, 27 distinct patterns were observed, 253 clinical isolates were grouped in 17 clusters (62.9%) and 10 isolates displayed an orphan pattern (37%). Based on an international spoligotype database, Haarlem I (85, 33.5%), Beijing (52, 20.5%), Central Asia (32, 12.1%), and EAI (21, 8.3%) were the major identified super families. Although, 76.9% of the Beijing genotypes and 100% of ST(253) strains (that was prevalent through former Soviet Union) were isolated from Afghan patients only. The linkage patterns between 30 Iranian and Afghan patients were observed. CONCLUSION: The study highlighted the epidemic potential of Haarlem I and Beijing genotypes among MDR-TB cases in Tehran territory. PMID- 16476484 TI - Myeloperoxidase and its contributory role in inflammatory vascular disease. AB - Myeloperoxidase (MPO), a heme protein abundantly expressed in polymorphonuclear neutrophils (PMN), has long been viewed to function primarily as a bactericidal enzyme centrally linked to innate host defense. Recent observations now extend this perspective and suggest that MPO is profoundly involved in the regulation of cellular homeostasis and may play a central role in initiation and propagation of acute and chronic vascular inflammatory disease. For example, low levels of MPO derived hypochlorous acid (HOCl) interfere with intracellular signaling events, MPO-dependent oxidation of lipoproteins modulates their affinity to macrophages and the vessel wall, MPO-mediated depletion of endothelial-derived nitric oxide (NO) impairs endothelium-dependent vasodilatation, and nitrotyrosine (NO(2)Tyr) formation by MPO sequestered into the vessel wall may affect matrix protein structure and function. Future studies are needed to further elucidate the significance of MPO in the development of acute and chronic vascular disease and to evaluate MPO as a potential target for treatment. PMID- 16476485 TI - From molecular action to physiological outputs: peroxisome proliferator-activated receptors are nuclear receptors at the crossroads of key cellular functions. AB - Peroxisome proliferator-activated receptors (PPARs) compose a family of three nuclear receptors which act as lipid sensors to modulate gene expression. As such, PPARs are implicated in major metabolic and inflammatory regulations with far-reaching medical consequences, as well as in important processes controlling cellular fate. Throughout this review, we focus on the cellular functions of these receptors. The molecular mechanisms through which PPARs regulate transcription are thoroughly addressed with particular emphasis on the latest results on corepressor and coactivator action. Their implication in cellular metabolism and in the control of the balance between cell proliferation, differentiation and survival is then reviewed. Finally, we discuss how the integration of various intra-cellular signaling pathways allows PPARs to participate to whole-body homeostasis by mediating regulatory crosstalks between organs. PMID- 16476487 TI - Severe melancholic depression is more vulnerable than non-melancholic depression to minor precipitating life events. AB - BACKGROUND: The present study examines the moderating role of global depression severity on the relation of melancholic versus non-melancholic depression to severe and non-severe levels of stress. METHOD: A community sample of 50 women with unipolar major depressive disorder, of which 54% met Research Diagnostic Criteria for melancholic depression, were interviewed regarding stressful life events experienced prior to onset. Events were coded as severe or non-severe based on the rigorous Bedford College contextual rating system. RESULTS: Greater severity of depression was related to a higher likelihood of a severely stressful event prior to onset only for women with non-melancholic major depression. By contrast, greater severity of depression was related to a higher likelihood of a non-severe, more minor, stressful event prior to onset only for women with melancholic major depression. LIMITATIONS: The present study was limited by its use of a female volunteer sample, which might not be entirely representative of the population of individuals with major depression. In addition, the study employed a cross-sectional design, which limits conclusions relating to the causal relation of stress to melancholic versus non-melancholic depression. CONCLUSIONS: Far from being autonomous of stress, individuals with severe melancholic depression may be especially sensitive to stress, such that their episodes are influenced by more minor stressors than those of individuals with non-melancholic depression. PMID- 16476486 TI - An improved test of neurological dysfunction following transient focal cerebral ischemia in rats. AB - The Adhesive Removal (sticky-tape) test is a commonly used test of somatosensory dysfunction following cerebral ischemia in rats. This test requires several days of pre-training prior to surgery, which can be time consuming. We present our results with an improved version of the sticky-tape test. Male Wistar rats were subjected to either sham surgery (n = 4) or right middle cerebral artery occlusion (rMCAo) using an intraluminal filament (n = 9), followed by a 10-day survival period. On post-operative days (POD) 1, 3, 7, and 10 animals underwent both the conventional sticky-tape test (CST) with measurement of the time to remove the stimulus (trs), as well as a modified sticky-tape test (MST), in which a non-removable tape sleeve was placed around the animal's paw. Time spent attending to this stimulus (tas) was recorded. Despite 3 days of pre-training, animals undergoing baseline CST still exhibited marked variability in pre operative baseline test performance (trs range 1-60s). In contrast, animals undergoing MST for the first time demonstrated nearly uniformly excellent performance (% tas range 91.5-98.5% of the 30s testing period). Although, affected (left) limb performance on both CST (6.8-fold increase in trs on POD 1 compared to baseline) and MST (100% decrease in tas on POD 1 compared to baseline) was markedly altered by rMCAo, CST performance declined bilaterally, and no significant differences in the ratio of affected (left) and unaffected (right) limb performance between sham-operated and rMCAo animals were observed at any time point. In contrast, the ratio of left to right performance on the MST was significantly different at all time points (P<0.01). In conclusion, we present a simple modification of the widely used Adhesive Removal test and provide evidence that this test can accurately assess neurological dysfunction in rodents, not only with minimal pre-training, but also with improved localization of the side of injury. PMID- 16476489 TI - Cloning and characterization of two genes encoding rainbow trout homologues of the IFITM protein family. AB - To identify genes connected with leucocyte function in rainbow trout a cDNA library derived from head kidney leucocytes exposed to phytohemagglutinin (PHA) was screened. A clone with homology to the previously described human interferon inducible transmembrane proteins (IFITM) was isolated. This clone was used to screen a trout genomic library identifying two trout IFITM genes. Both genes are composed of two exons, in common with human IFITM genes. The promoter regions of both genes were examined and found to contain interferon, immune and stress related transcription factor binding sites. Transcripts for both genes were found in most tissues examined. Trout IFITM1 mRNA levels were highest in head kidney, but present in all tissues studied. IFITM2 mRNA levels were highest in gill, heart and liver, and absent in head kidney and blood. In response to conditions mimicking viral infection, transcription of the trout IFITM genes was elevated in a trout spleen macrophage cell line (RTS11) over the examined timecourse (72 h). This study suggests that the organization and transcription of the IFITM genes is well conserved between human and rainbow trout. PMID- 16476488 TI - Immune responses to commercial equine vaccines against equine herpesvirus-1, equine influenza virus, eastern equine encephalomyelitis, and tetanus. AB - Horses are commonly vaccinated to protect against pathogens which are responsible for diseases which are endemic within the general horse population, such as equine influenza virus (EIV) and equine herpesvirus-1 (EHV-1), and against a variety of diseases which are less common but which lead to greater morbidity and mortality, such as eastern equine encephalomyelitis virus (EEE) and tetanus. This study consisted of two trials which investigated the antigenicity of commercially available vaccines licensed in the USA to protect against EIV, EHV-1 respiratory disease, EHV-1 abortion, EEE and tetanus in horses. Trial I was conducted to compare serological responses to vaccines produced by three manufacturers against EIV, EHV-1 (respiratory disease), EEE, and tetanus given as multivalent preparations or as multiple vaccine courses. Trial II compared vaccines from two manufacturers licensed to protect against EHV-1 abortion, and measured EHV-1 specific interferon-gamma (IFN-gamma) mRNA production in addition to serological evidence of antigenicity. In Trial I significant differences were found between the antigenicity of different commercial vaccines that should be considered in product selection. It was difficult to identify vaccines that generate significant immune responses to respiratory viruses. The most dramatic differences in vaccine performance occurred in the case of the tetanus antigen. In Trial II both vaccines generated significant antibody responses and showed evidence of EHV-1-specific IFN-gamma mRNA responses. Overall there were wide variations in vaccine response, and the vaccines with the best responses were not produced by a single manufacturer. Differences in vaccine performance may have resulted from differences in antigen load and adjuvant formulation. PMID- 16476490 TI - Molecular cloning and characterization of cDNA encoding CD11b of cattle. AB - CD18, the common beta subunit of beta2-integrins, associates with four distinct alpha chains to give rise to four different beta2-integrins: CD11a/CD18 (LFA-1), CD11b/CD18 (Mac-1), CD11c/CD18 (CR4), and CD11d/CD18. Previously, we and others showed that CD18 of LFA-1 serves as a receptor for Mannheimia haemolytica leukotoxin (Lkt). Level of expression of Mac-1 is higher than that of LFA-1 and other beta2-integrins on polymorphonuclear leukocytes (PMNs), which constitute the leukocyte subset most susceptible to Lkt. Hence, it is likely that CD18 of Mac-1 also mediates Lkt-induced cytolysis. Co-expression of CD11b and CD18 of cattle on Lkt-resistant cells is necessary to irrefutably demonstrate the role of Mac-1 in Lkt-induced cytolysis. This approach is hindered by lack of availability of complete sequence of cattle CD11b. Therefore, in this study, we cloned and sequenced the full length cDNA encoding cattle CD11b. The 3459 bp cDNA of cattle CD11b encodes a polypeptide of 1152 amino acids. The deduced amino acid sequence of CD11b of cattle exhibits 75% identity to that of humans and chimpanzees, 74% identity to that of dogs, and 70% identity to that of mice and rats. Availability of cattle CD11b cDNA should facilitate the elucidation of Lkt-receptor interactions in cattle and other species. PMID- 16476491 TI - Effects of antigen and recombinant porcine cytokines on pig dendritic cell cytokine expression in vitro. AB - To evaluate variables influencing in vitro immune response induction, pig monocyte-derived DCs (moDCs) were treated with putative type-1 and type-2 antigens (Ags, killed Mycobacterium tuberculosis (Mtb) and hen egg white lysozyme (HEWL)) and recombinant porcine cytokines (IL-6, IL-10, IL-12, IFN-gamma and TNF alpha). Responses were measured as moDC cytokine mRNA expression. Treatment of moDCs with HEWL increased IL-13 but not IL-12, IFN-gamma or IL-10 mRNA, suggesting a DC2 phenotype. Addition of TNF-alpha, IFN-gamma or IL-12 to HEWL treated moDCs increased IL-12p35 and reduced IL-13 mRNA; suggesting a DC1 phenotype. Mtb increased moDC IL-12p35, IFN-gamma and to a lesser extent IL-13 mRNA. This DC1 bias was enhanced by TNF-alpha, IFN-gamma or IL-12, which increased IL-12p35 and to a lesser extent IL-10 mRNA but reduced IL-13 mRNA. Addition of IL-10 to Mtb-pulsed moDCs reduced IL-12p35, IFN-gamma and IL-13, but increased IL-10 mRNA, suggesting diversion from DC1 to DC2. Thus porcine moDCs treated with Ag and/or cytokines alter moDC cytokine expression confirming their likely ability to initiate and steer acquired immune response. PMID- 16476492 TI - The equine immune response to equine herpesvirus-1: the virus and its vaccines. AB - Equine herpesvirus-1 (EHV-1) is an alphaherpesvirus which infects horses, causing respiratory and neurological disease and abortion in pregnant mares. Latency is established in trigeminal ganglia and lymphocytes. Immunity to EHV-1 lasts between 3 and 6 months. Current vaccines, many of which contain inactivated virus, have reduced the incidence of abortion storms in pregnant mares but individual animals, which may be of high commercial value, remain susceptible to infection. The development of effective vaccines which stimulate both humoral and cellular immune responses remains a priority. Utilising data generated following experimental and field infections of the target species, this review describes the immunopathogenesis of EHV-1 and the interaction between the horse's immune system and this virus, both in vivo and in vitro, and identifies immune responses, highlighting those which have been associated with protective immunity. It then goes on to recount a brief history of vaccination, outlines factors likely to influence the outcome of vaccine administration and describes the immune response stimulated by a selection of commercial and experimental vaccines. Finally, based on the available data, a rational strategy designed to stimulate protective immune responses by vaccination is outlined. PMID- 16476493 TI - Genotyping of Toll-like receptor 4, myeloid differentiation factor 2 and CD-14 in the horse: an investigation into the influence of genetic polymorphisms on the LPS induced TNF-alpha response in equine whole blood. AB - The inter- and intra-species differences in the response to lipopolysaccharides (LPS) are well recognised in mammalian species. It has been hypothesized that these differences can be attributed to genetic polymorphisms in the components involved in LPS signal transduction. These components include the cluster of differentiation factor 14 (CD-14), a membrane bound protein on the surface of mononuclear cells that recognises LPS and a receptor complex consisting of Toll like receptor-4 (TLR-4) and myeloid differentiation factor-2 (MD-2). Sequencing of these three proteins in humans and mice revealed that all three are susceptible to polymorphic alterations, influencing the response to LPS. Previous experiments in the horse showed large inter-individual variations in the response to LPS. With the aim to assess this inter-individual variation, we performed a whole blood assay in 10 healthy horses as a functional assay to study the responsiveness to LPS. In 3 out of the 10 horses, LPS-induced TNF-alpha production was significantly lower compared to the overall mean. Subsequently the entire cDNA sequence encoding for the TLR-4, MD-2 and CD-14 protein was documented for each horse. Although mutations were observed in the sequence of TLR-4, these could not be related to an altered response to LPS in the concentration used in this study, as determined in the whole blood assay. Despite the various mutations found in the TLR-4 receptor protein, no alterations could be found in either the MD-2 or CD-14 gene, which are obviously more conserved structures. PMID- 16476494 TI - A brief history of cannabinoid and endocannabinoid pharmacology as inspired by the work of British scientists. AB - British scientists have played a leading role in the long history of cannabinoid and endocannabinoid research. Such research has progressed from the first crucial evaluation of the medicinal properties of Cannabis sativa in the Western world to pioneering studies of the chemical constituents of this plant, the development of in vitro biological assays to study cannabinoids, the identification of the mechanism of action of cannabinoids, the discovery of endocannabinoids and the assessment of their therapeutic implications. Stemming from the many innovative ideas and achievements of these researchers, I provide a personal view of where these studies have led us thus far and where they are likely to take us in the future. PMID- 16476495 TI - A modelling study of the potential influence of low dose hypersensitivity on radiation treatment planning. AB - BACKGROUND AND PURPOSE: Low dose hyper-radiosensitivity (HRS) has been observed in both normal tissues and tumours. This modelling study explores the possible impact of HRS on radiation treatment planning. PATIENTS AND METHODS: The interplay between volume-effect and HRS was studied in an idealized comparison of partial versus whole organ irradiation. In the further studies, CT scans of three previously scanned patients were used to estimate normal tissue complication probability (NTCP) for the kidneys after a conformal and a conventional treatment plan with and without consideration of HRS. RESULTS: Idealized treatment plans were compared as pairs of a conventional and a conformal plan both treating the same target volume to the same dose per fraction. Contour maps of the difference in NTCP between paired plans showed a strong dependence on the magnitude of both the volume effect and the HRS effect. For more clinically realistic treatment plans with NTCP calculated for the kidney, the balance between the sparing due to the LQ effect and the increased sensitivity due to the HRS effect was dependent on both the dose distribution and the fractionation. CONCLUSIONS: HRS may potentially affect radiotherapy treatment planning and the relative importance of HRS is larger in a tissue or organ with a pronounced volume effect. If HRS is expressed in some normal tissues or organs, this could offset much of the sparing predicted by the LQ formalism. However, in some clinical situations the NTCP calculated with correction for HRS may still be lower than the NTCP calculated from the uncorrected physical doses. PMID- 16476496 TI - Molecular imaging to improve radiotherapy. PMID- 16476497 TI - Improved extracellular phytase activity in Saccharomyces cerevisiae by modifications in the PHO system. AB - Myo-inositol hexaphosphate (IP6, phytate) is a potent anti-nutritional compound occurring in many plant-based staple foods, limiting the bioavailability of important nutrients such as iron and zinc. The objective of the present study was to investigate different strategies to achieve high and constitutive extracellular IP6 degradation by Baker's yeast, Saccharomyces cerevisiae. By deleting either of the genes PHO80 and PHO85, encoding negative regulators of the transcription of the repressible acid phosphatases (rAPs), the IP6 degradation became constitutive, and the biomass specific IP6 degradation was increased manyfold. In addition, the genes encoding the transcriptional activator Pho4p and the major rAP Pho5p were overexpressed in both a wild-type and a pho80delta strain, yielding an additional increase in IP6 degradation. It has previously been proved possible to increase human iron bioavailability by degradation of IP6 using microbial phytase. A high-phytase S. cerevisiae strain, without the use of any heterologous DNA, may be a suitable organism for the production of food-grade phytase and for the direct use in food production. PMID- 16476498 TI - The origin of viruses and their possible roles in major evolutionary transitions. AB - Viruses infecting cells from the three domains of life, Archaea, Bacteria and Eukarya, share homologous features, suggesting that viruses originated very early in the evolution of life. The three current hypotheses for virus origin, e.g. the virus first, the escape and the reduction hypotheses are revisited in this new framework. Theoretical considerations suggest that RNA viruses may have originated in the nucleoprotein world by escape or reduction from RNA-cells, whereas DNA viruses (at least some of them) might have evolved directly from RNA viruses. The antiquity of viruses can explain why most viral proteins have no cellular homologues or only distantly related ones. Viral proteins have replaced the ancestral bacterial RNA/DNA polymerases and primase during mitochondrial evolution. It has been suggested that replacement of cellular proteins by viral ones also occurred in early evolution of the DNA replication apparatus and/or that some DNA replication proteins originated directly in the virosphere and were later on transferred to cellular organisms. According to these new hypotheses, viruses played a critical role in major evolutionary transitions, such as the invention of DNA and DNA replication mechanisms, the formation of the three domains of life, or else, the origin of the eukaryotic nucleus. PMID- 16476499 TI - Microenvironmental pH and microviscosity inside pH-controlled matrix tablets: an EPR imaging study. AB - Incorporation of pH modifiers is a commonly used strategy to enhance the dissolution rate of weakly basic drugs from sustained release solid dosage forms. Electron paramagnetic resonance imaging (EPRI) was applied to spatially monitor pH(M) and the rotational correlation time (tau(R)), a parameter which is closely related to the surrounding microviscosity inside HPMC (hydroxypropylmethylcellulose) matrix tablets. Fumaric, citric, and succinic acid were employed as pH modifiers. 4-(methylamino)-2-ethyl-5,5-dimethyl-4-pyridine-2 yl-2,5-dihydro-1H-imidazole-1-oxyl (MEP) was used as spin label. Fumaric and citric acid reduced the pH(M) to equal extents in the initial phase. With the progress of hydration, the more soluble citric acid diffused out from the tablet resulting in an increase in pH(M), originating at the outer layers. In contrast, fumaric acid maintained a constantly reduced pH(M) inside the entire tablet. Due to its lower acidic strength, succinic acid did not reduce the pH(M) as effectively as the other pH modifiers used. The more water-soluble acids stimulated the water penetration into the matrix system, thereby rapidly decreasing tau(R). Once the matrix tablets were hydrated, the included pH modifiers influenced tau(R) insignificantly. EPRI, a novel approach for monitoring pH(M) and tau(R) non-invasively and spatially resolved, was used successfully for the optimization of an pH-controlled formulation. PMID- 16476500 TI - Fabrication and characterization of microfluidic probes for convection enhanced drug delivery. AB - Convection enhanced drug delivery (CED) is a promising therapeutic method for treating diseases of the brain by enhancing the penetration of drugs. Most controlled release delivery methods rely on diffusion from a source to transport drugs throughout tissue. CED relies on direct infusion of drugs into tissue at a sufficiently high rate so that convective transport of drug is at least as important as diffusive transport. This work describes the fabrication and characterization of microfluidic probes for CED protocols and the role diffusion plays in determining penetration. Microfluidic channels were formed on silicon substrates by employing a sacrificial photoresist layer encased in a parylene structural layer. Flow in the microchannels was characterized by applying constant upstream pressures from 35 to 310 kPa, which resulted in flow rates of 0.5-4.5 microL/min. The devices were used to infuse Evans Blue and albumin in hydrogel brain phantoms. The results of these infusions were compared to a simple convection-diffusion model for infusions into porous media. In vivo infusions of albumin were performed in the gray matter of rats at upstream pressures of 35, 70, and 140 kPa. The microfabricated probes show reduced evidence of backflow along the device-tissue interface when compared with conventional needles used for CED. PMID- 16476502 TI - The relationship between hospital capacity characteristics and emergency department volumes in Korea. AB - OBJECTIVE: To describe the level of emergency department (ED) volumes according to the hospital characteristics and to identify the relationship between hospital capacity characteristics and ED volumes in Korea. METHOD: A survey was conducted to acquire information on the ED, its' hospital (facility, personnel, equipment), and the number of ED patients, as part of the National Emergency Medical Centers Assessment Program. Data from 106 nation-wide LEMCs were used. Multiple regression analysis was performed to determine the hospital capacity characteristics related with ED volumes. RESULTS: The number of ED patients differed according to bed size, nurse staffing, residency training program, and the availability of emergency care-related equipment of the hospital. In the multiple regression analysis, the significant factors which explained the ED volumes were nurse staffing, inpatients per bed, and the population in the area where hospitals are located. The hospitals that were nurse staffing level 2, with more inpatients per bed and larger population of the service area, had more ED patients. CONCLUSIONS: With the service area population, the ED volumes significantly related with nurse staffing and inpatients per bed. These could be used as one of criteria to designate a LEMC. PMID- 16476501 TI - Effect of stress management training on glycemic control in patients with type 1 diabetes. AB - AIMS: In this study, the effect of stress management training on glycemic control has been investigated in type 1 diabetic patients. METHODS: The participants were 60 type 1 diabetic patients (aged 16-30). 30 Subjects attended in 3-month stress management training classes during which the prescribed insulin remained constant, but the remainder 30 ones did not. HbA1 from all patients were measured before and after the intervention. Besides, in order to assess the ways of coping, every patient completed a questionnaire and the scores were compared between two groups. RESULTS: Trained patients showed significantly improved ways of coping. HbA1 changed from 11.7+/-2.9 and 10.9+/-2.1 before training to 8.5+/ 1.7 and 10.3+/-2.1 after intervention in trained and control groups respectively and the changes were significant in study group (P<0.001). In addition, the difference between means of HbA1 of two groups was statistically significant at the end of the study (P<0.001). CONCLUSIONS: Results show a clinically significant beneficial effect of stress management training on glycemic control among type 1 diabetic patient. It is recommended to consider this type of training as an addition to the treatment program in type 1 diabetic patient. PMID- 16476503 TI - Distributed computing methodology for training neural networks in an image-guided diagnostic application. AB - Distributed computing is a process through which a set of computers connected by a network is used collectively to solve a single problem. In this paper, we propose a distributed computing methodology for training neural networks for the detection of lesions in colonoscopy. Our approach is based on partitioning the training set across multiple processors using a parallel virtual machine. In this way, interconnected computers of varied architectures can be used for the distributed evaluation of the error function and gradient values, and, thus, training neural networks utilizing various learning methods. The proposed methodology has large granularity and low synchronization, and has been implemented and tested. Our results indicate that the parallel virtual machine implementation of the training algorithms developed leads to considerable speedup, especially when large network architectures and training sets are used. PMID- 16476504 TI - Diagnostic agreement in the histopathological evaluation of lung cancer tissue in a population-based case-control study. AB - Only few studies have compared the agreement of histological lung carcinoma diagnosis of a population-based case series and an independent pathology review. We analyzed data of our population-based lung cancer case-control study to determine the agreement in the histopathological evaluation of lung cancer. Six hundred and sixty-eight out of 1004 interviewed male and female lung cancer cases were histologically evaluated according to the 1981 WHO classification by regional pathologists and a central pathologist who was blinded to the evaluations of the regional pathologists. The observed agreement was 0.65 with kappa = 0.54 (95% CI: 0.49-0.58). It was highest for small-cell carcinoma (0.94; kappa = 0.82) and lower for squamous-cell carcinoma (0.81; kappa = 0.58) and adenocarcinoma (0.81; kappa = 0.55). Agreement was slightly higher among women than men. The observed agreement among non-smoking cases was 58% as compared to 67% heavy smoking cases. The moderate agreement for squamous-cell and adenocarcinoma complicates epidemiological studies that address these histological subtypes. PMID- 16476505 TI - Association of TNF-alpha polymorphism with susceptibility to and severity of non small cell lung cancer. AB - BACKGROUND: Genetic polymorphisms in the promoter region of the tumor necrosis factor-alpha (TNF-alpha) gene are involved in the regulation of expression levels and have been associated with various inflammatory and malignant conditions. We have investigated two polymorphisms in the promoter region of the TNF-alpha gene (-308 G/A and -238 G/A) for their role in the susceptibility to and severity of non-small cell lung cancer (NSCLC), by means of an allelic association study. METHODS: Using a case-control study design, lung cancer patients (n = 202) and appropriate age- and sex-matched controls recruited from the health check-up unit (n = 205) were subjected to genotype analysis for these polymorphisms, using a high-throughput allelic discrimination method. RESULTS: Genotype was analyzed using the polymerase chain reaction-restriction fragment length polymorphism technique with genomic DNA isolated from peripheral blood lymphocytes. Overall, the distribution of the genotype frequencies of TNF-alpha-308 A/G and -238 A/G were significantly different between the lung cancer patients and the healthy controls, and also different between patients with lung cancers of various stages (p < 0.0001). Logistic regression analysis revealed that higher odds ratios (ORs) for lung cancer were seen for individuals with TNF-alpha-308 AA/GA genotypes against GG genotype (an OR of 3.75, 95% CI 2.38-5.92, p < 0.0001), and lower ORs were seen for individuals with TNF-alpha-238 AA/GA genotypes against GG genotype (an OR of 0.26, 95% CI 0.13-0.50, p < 0.0001). The patients carrying a homologous AA or heterologous GA genotype at TNF-308 (p = 0.017), or a homologous GG genotype at TNF-238 (p = 0.001), had a tendency to advanced disease. CONCLUSIONS: A significant association between the 308 G/A and 238 G/A polymorphisms in the promoter region of TNF-alpha and the susceptibility to lung cancer was demonstrated. Also, these two polymorphisms were associated with the severity of lung cancer. The -308 A allele has a promotive effect for lung cancer development and progression, whereas the -238 A allele has a protective function against lung cancers. PMID- 16476506 TI - Upscaling heterogeneity in aquifer reactivity via the exposure-time concept: inverse model. AB - A novel inverse technique is proposed to quantitatively characterize macroscopic variability in aquifer reactivity in a Lagrangian representation. Reactivity heterogeneity is expressed in terms of distributions of flux over cumulative time of exposure of the solution to reactive surface area, termed here 'cumulative reactivity'. In cases involving single aqueous species the combined effects of physical and reactivity heterogeneity on reactive solute transport can often be established and further investigated through joint distributions of flux over travel time and cumulative reactivity. The inverse technique requires the breakthrough curve of a passive tracer to determine the distribution of flux over travel time, and additional breakthrough curves of reactive tracers provide additional moments of the distribution of flux over cumulative reactivity given travel time. Thus breakthroughs of one passive and two reactive tracers can provide the mean and variance of the distribution of flux over cumulative reactivity. This Lagrangian characterization is achieved with knowledge of the types of reactive surfaces present, but not their spatial locations. The distributions can subsequently be applied via forward modeling using the same technique to predict breakthrough curves of other solutes undergoing first-order reactions in similar physically and chemically heterogeneous configurations. PMID- 16476507 TI - Modifications of the human urocortin 2 peptide that improve pharmacological properties. AB - Recently, we demonstrated that the corticotropin releasing factor 2 receptor agonist, urocortin 2, demonstrated anti-atrophy effects in rodent skeletal muscle atrophy models. Compared to other CRF2R agonists however, the in vivo pharmacological potency of urocortin 2 is poor when it is administered by continuous subcutaneous infusion. Therefore, we attempted to modify the structure of urocortin 2 to improve in vivo efficacy when administered by subcutaneous infusion. By substituting amino acid residues in the linker region of urocortin 2 (residues 22-32), we have demonstrated improved in vivo potency without altering selectivity, probably through reduced CRFBP binding. In addition, attempts to shorten urocortin 2 generally resulted in inactive peptides, demonstrating that the 38 amino acid urocortin 2 peptide is the minimal pharmacophore. PMID- 16476508 TI - Differential BBB interactions of three ingestive peptides: obestatin, ghrelin, and adiponectin. AB - Endogenous compounds, including ingestive peptides, can interact with the blood brain barrier (BBB) in different ways. Here we used in vivo and in vitro techniques to examine the BBB permeation of the newly described satiety peptide obestatin. The fate of obestatin in blood and at the BBB was contrasted with that of adiponectin. By the sensitive multiple time-regression method, obestatin appeared to have an extremely fast influx rate to the brain whereas adiponectin did not cross the BBB. HPLC analysis, however, showed the obestatin result to be spurious, reflecting rapid degradation. Absence of BBB permeation by obestatin and adiponectin was in contrast to the saturable transport of human ghrelin reported previously. As a positive control, ghrelin showed saturable binding and endocytosis in RBE4 cerebral microvessel endothelial cells. By comparison, obestatin lacked specific binding and endocytosis, and the small amount internalized showed rapid intracellular degradation before the radioactivity was released by exocytosis. The differential interactions of obestatin, adiponectin, and ghrelin with the BBB illustrate their distinctive physiological interactions with the CNS. PMID- 16476510 TI - Inducing systemic and mucosal immune responses to B-T construct of F1 antigen of Yersinia pestis in microsphere delivery. AB - Plague is a zoonotic disease caused by Yersinia pestis, an etiological agent of pneumonic and bubonic plague. There is a need for an improved plague vaccine that may overcome the limitation of presently available whole cell vaccine. An alternative approach described here, is the use of protective epitopes from immunodominant antigen of Y. pestis. One such antigen is the F1 antigen, a major envelope and virulent protein that possess antiphagocytic and anti-microbial properties. The present study was aimed to develop a peptide-based vaccine, based upon the constructs made between B and T cell epitopes of F1 antigen of Y. pestis. The immunogenicity, IgG subclass pattern, affinity, avidity and in vivo protective efficacy of the antibodies generated for different B-T constructs were studied in murine model using microsphere as the delivery vehicle. The mode of immunization was both intranasal and intramuscular, with single and multiple doses of immunization, respectively. Intranasal immunization generated consistent high titre and long lasting immune response both for IgG and IgA in sera and sIgA in washes while intramuscular route generated peak IgG levels in sera only. The IgG isotypic levels pattern showed higher IgG2a/IgG2b levels in intranasal route while mixed isotypic levels of IgG1, IgG2a/IgG2b were observed in intramuscular route. The affinity and relative avidity of antibodies showed best results with intranasal route as compared to the intramuscular route. The specific activity measurement (IgG/IgA content) in sera and washes were well correlated with the antibody levels. Finally, in vivo protective studies showed that B1T1 and B2T1 conjugates protected the mice till day 15 while rest of the conjugates showed poor protection. PMID- 16476512 TI - Sub-ungual epidermoid cyst. PMID- 16476511 TI - Resuspension and redistribution of radionuclides during grassland and forest fires in the Chernobyl exclusion zone: part II. Modeling the transport process. AB - To predict parameters of radionuclide resuspension, transport and deposition during forest and grassland fires, several model modules were developed and adapted. Experimental data of controlled burning of prepared experimental plots in the Chernobyl exclusion zone have been used to evaluate the prognostic power of the models. The predicted trajectories and elevations of the plume match with those visually observed during the fire experiments in the grassland and forest sites. Experimentally determined parameters could be successfully used for the calculation of the initial plume parameters which provide the tools for the description of various fire scenarios and enable prognostic calculations. In summary, the model predicts a release of some per thousand from the radionuclide inventory of the fuel material by the grassland fires. During the forest fire, up to 4% of (137)Cs and (90)Sr and up to 1% of the Pu isotopes can be released from the forest litter according to the model calculations. However, these results depend on the parameters of the fire events. In general, the modeling results are in good accordance with the experimental data. Therefore, the considered models were successfully validated and can be recommended for the assessment of the resuspension and redistribution of radionuclides during grassland and forest fires in contaminated territories. PMID- 16476513 TI - Dorsal capsulodesis for scapholunate instability--effect on patient disability and wrist pain. AB - Twenty cases of scapho-lunate instability have been reviewed, to determine the effect of dorsal capsulodesis on disability and wrist pain. The average follow-up was 41 (range 13-63) months. There was a significant improvement in the pre operative Patient Rated Wrist Evaluation score from 108 (range 18-150) to a postoperative score of 60 (range 0-132). All wrist movements and grip strength were reduced postoperatively but only the reductions in flexion and radial deviation were statistically significant. Seventeen (85%) patients were satisfied with the operation. One patient remained unemployed after surgery. Five patients returned to their normal jobs. This study shows that dorsal capsulodesis should remain an important option in the treatment of scapholunate instability. PMID- 16476514 TI - Community unemployment and immigrants' health in Montreal. AB - This research examines the relationship between community unemployment and the physical and mental health of immigrants in comparison to non-immigrants in Montreal under the hypothesis that high unemployment in the community may generate more negative effects on the health of immigrants than on non immigrants. Possible gender differences in these associations are also examined. Montreal residents were studied via multilevel analysis, using both individual survey data and neighbourhood data from 49 police districts. Individual-level data were excerpted from a 1998 health survey of Montreal residents, while neighbourhood data originated from survey data collected in the 49 Montreal police districts and the 1996 Canadian Census. The associations between community unemployment and self-rated health, psychological distress and obesity are examined, and hypotheses regarding the modifying mechanisms via which male and female immigrants may run a greater risk of poor health than non-immigrants when living in areas of high unemployment were tested. Between neighbourhoods, variations in the three health outcomes were slight, and differences in health were not associated with differences in community unemployment. The associations between community unemployment and health varied according to immigration status. At the individual level, immigrants do not differ from non-immigrants with respect to the three health indicators, except that second-generation males are slightly heavier. However, when living in areas of high unemployment, immigrants tend to report poor physical and mental health in comparison to non-immigrants. Among first-generation immigrants, community unemployment was associated with psychological distress. Among second-generation immigrants, the probability of obesity and poor self-rated health increased significantly for those living in areas with high unemployment, but these associations reached statistical significance only for men. Findings among first-generation immigrants are interpreted with respect to the effects of possible discrimination in areas with low job availability. Among second-generation men, poor physical health and obesity may be the result of poor health habits stemming from perceived lack of life opportunities. PMID- 16476515 TI - Out of the dissecting room: news media portrayal of human anatomy teaching and research. AB - Radical changes in medical research and education have recently led to a number of innovative developments in terms of how human anatomy is represented and understood. New ways of introducing medical students to anatomy (including living anatomies and virtual simulations) have provoked widespread debate, with discussion of their relative merits compared to more traditional approaches that use cadaveric dissection. Outside the field of medicine, in the wider public sphere, the practice of anatomical study may often seem mysterious. The dissemination of news on anatomy, we contend, is central to the question of how medical researchers and educators engage with the public. Our analysis of news media coverage in the UK demonstrates that news-making, by giving prominence to certain facts, themes and images, serves to mask issues about anatomy and its practices that need debate. We examine the ways in which news media, through processes of selection and the 'framing' of issues, may perform an agenda-setting role. We draw attention to the use of positive 'awe and amazement' frames including 'miracles of modern science', 'medical heroes', and 'gifts of life', alongside more negative 'guts and gore' coverage including 'Frankenstein', 'Brave New World' and 'Rape of the Body' frames that concentrate on high profile scandals associated with the use and misuse of human bodies, tissues and parts. We also highlight the selective use of commentaries from members of the medical profession, which are more prevalent in positive 'awe and amazement' stories than in stories with negative coverage. We conclude by arguing for greater collaboration between journalists on the one hand, and medical educators and researchers on the other, in the making of news in order to provide portrayals of anatomy which bear a closer relationship to the everyday reality of professional work. PMID- 16476516 TI - How social context impacts on women's fears of childbirth: a Western Australian example. AB - This paper addresses the limited sociological understanding of the phenomena of childbirth fear using data from a qualitative research project conducted in Western Australia. This qualitative study used an exploratory descriptive design, with 22 women identified as being fearful of birth participating in an in-depth interview. Data analysis using the method of constant comparison revealed that social context, explored within the framework of the medicalisation of childbirth, and the intervening circumstances in which the women gave birth, impacted on how and why they experienced fear. As such, this paper argues that fear of childbirth has social as well as personal dimensions and is both a prospective and retrospective phenomena. The analysis identified prospective fear as both social and personal. The social dimensions were labelled as 'fear of the unknown', 'horror stories' and 'general fear for the well-being of the baby'. Personal dimensions included the 'fear of pain', 'losing control and disempowerment' and 'uniqueness of each birth'. Retrospective fear was exclusively personal and was clustered around the themes of 'previous horror birth' and 'speed of birth'. The analysis also revealed two central factors that mediated against childbirth fear: positive relationships formed with midwives, and the support women received from their informal network. Understanding and unpacking the dimensions of women's childbirth fear, and understanding the nature of relationships that mediate women's fear, provides health care professionals with information on which to base potential intervention strategies and support women in ways that lessen rather than heighten their fear. PMID- 16476517 TI - Fear extinction in rats: implications for human brain imaging and anxiety disorders. AB - Fear extinction is the decrease in conditioned fear responses that normally occurs when a conditioned stimulus (CS) is repeatedly presented in the absence of the aversive unconditioned stimulus (US). Extinction does not erase the initial CS-US association, but is thought to form a new memory. After extinction training, extinction memory competes with conditioning memory for control of fear expression. Deficits in fear extinction are thought to contribute to post traumatic stress disorder (PTSD). Herein, we review studies performed in rats showing that the medial prefrontal cortex plays a critical role in the retention and expression of extinction memory. We also review human studies indicating that prefrontal areas homologous to those critical for extinction in rats are structurally and functionally deficient in patients with PTSD. We then discuss how findings from rat studies may allow us to: (1) develop new fear extinction paradigms in humans, (2) make specific predictions as to the location of extinction-related areas in humans, and (3) improve current extinction-based behavioral therapies for anxiety disorders. PMID- 16476518 TI - Failed hypospadias repair presenting in adults. AB - OBJECTIVES: To evaluate the results of one-stage and multistage urethroplasty in adults with complications following multiple failed hypospadias repairs. METHODS: Sixty adults underwent reconstructive surgery of the following complications after hypospadias repair: stricture (34), residual hypospadias (26), fistula (18), meatal stenosis (11), penile curvature (9), hair (4), diverticula (2), and stone (1). A total of 36% of the patients had one complication and 64% had two or more complications. Twenty-nine patients underwent one-stage repair with buccal or skin grafts or direct repair, and 31 underwent multistage repairs with buccal or skin grafts. RESULTS: Mean follow-up was 33.8 mo. Of the 60 cases, 45 (75%) had a final successful outcome and 15 (25%) failed. One-stage repair provided 24 (82.7%) successes and 5 (17.3%) failures. Multistage repair provided 21 (67.7%) successes and 10 (32.3%) failures. Buccal mucosa grafts provided 81% of success in one-stage procedures and 82.3% in multistage procedures. Penile skin grafts provided 80% of success in one-stage procedures and 50% in multistage procedures. CONCLUSIONS: Adults with complications following childhood hypospadias repair are still a difficult population to treat with a high failure rate for reoperative surgery. PMID- 16476519 TI - Antitumor activity of the antimicrobial peptide magainin II against bladder cancer cell lines. AB - OBJECTIVE: Magainin II belongs to a family of antimicrobial peptides and has been shown to exhibit antibiotic activity in a wide range of organisms. Recent studies have also reported a significant antitumor effect of magainin II against various cancer cell lines and tumor mice models. In this study, we evaluated the cytotoxic and antiproliferative potency of magainin II in bladder tumor cells and normal fibroblasts. METHODS: The antiproliferative and cytotoxic effect of magainin II was quantified by colorimetric WST-1-, bromodeoxyuridine (BrdU)-, and lactic dehydrogenase (LDH) assays in three bladder cancer cell lines (RT4, 647V, and 486P) and in the murine fibroblast cell line 3T3 as well as in a primary culture from human fibroblasts. The median inhibitory concentration (IC50) values were determined for each assay, representing the concentration at which cell viability was reduced by 50%. Scanning electron microscopy (SEM) was used to visualize the morphologic effects of magainin II on bladder tumor cells and fibroblasts. RESULTS: Magainin II inhibited cell proliferation of bladder cancer cells in a dose-dependent manner. The average IC50 of magainin II against all bladder cancer cell lines was 198.1 microM (range, 52.4-484.03 microM) for the WST-1 assay and 75.2 microM (range, 31.0-135.3 microM) for the BrdU assay. The normal murine and human fibroblast cell lines were not affected by magainin II and their IC50 could not be determined at the concentrations of magainin II tested. LDH release was increased in all bladder tumor cell lines in the presence of magainin II, whereas normal fibroblasts showed no cell lysis. SEM demonstrated lethal membrane perforation by peptide pore formation in bladder cancer cells, but not in fibroblasts. CONCLUSION: Magainin II peptide exerts cytotoxic and antiproliferative efficacy by pore formation in bladder cancer cells but has no effect on normal murine or human fibroblasts. Magainin II may offer a novel therapeutic strategy in the treatment of bladder cancer with potentially low cytotoxic effects on normal cells. PMID- 16476521 TI - Centaureidin promotes dendrite retraction of melanocytes by activating Rho. AB - Melanosomes synthesized within melanocytes are transferred to keratinocytes through dendrites, resulting in a constant supply of melanin to the epidermis, and this process determines skin pigmentation. During screening for inhibitors of melanosome transfer, we found a novel reagent, centaureidin, that induces significant morphological changes in normal human epidermal melanocytes and inhibits melanocyte dendrite elongation, resulting in a reduction of melanosome transfer in an in vitro melanocyte-keratinocyte co-culture system. Since members of the Rho family of small GTP-binding proteins act as master regulators of dendrite formation, and activated Rho promotes dendrite retraction, we studied the effects of centaureidin on the small GTPases. In in vitro binding assay, centaureidin activated Rho and furthermore, a Rho inhibitor (C. botulinum C3 exoenzyme), a Rho kinase inhibitor (Y27632) and a small GTPase inhibitor (Toxin B) blocked dendrite retraction induced by centaureidin. These results suggest centaureidin could act via the Rho signaling pathway, and it may directly or indirectly activate Rho. Thus, centaureidin appears to inhibit dendrite outgrowth from melanocytes by activating Rho, resulting in the inhibition of melanosome transfer from melanocytes to keratinocytes. PMID- 16476520 TI - Actions of 5alpha-reductase inhibitors on the epididymis. AB - Testosterone is converted to the more biologically active androgen, dihydrotestosterone (DHT), by steroid 5alpha-reductase. Two isozymes of 5alpha reductase, types 1 and 2, are abundantly expressed in the epididymis. DHT is the androgen found in the nuclei of epididymal cells and is essential for the maturation of spermatozoa. Thus, one approach to block androgen action in the epididymis is to inhibit DHT formation. Several compounds have been reported to inhibit either one or both forms of 5alpha-reductase in many tissues. The first commercially available inhibitor of 5alpha-reductase, finasteride, has a predominant effect on the type 2 isozyme, while more recently developed agents, such as dutasteride, PNU157706 and FK143, act as dual inhibitors. We found that the treatment of adult rats with such agents results in pronounced effects on the expression of genes essential to the formation of the optimal luminal microenvironment that is required for proper sperm maturation. Furthermore, drug treatment caused a significant decrease in the percentage of progressively motile and morphologically normal spermatozoa in the cauda epididymides. Mating females to treated males resulted in fewer successful pregnancies and a higher rate of pre-implantation loss. Thus, there may be a role for dual 5alpha-reductase inhibitors as potential components of a male contraceptive. PMID- 16476522 TI - Full thickness skin loss following chemical tattoo removal. PMID- 16476523 TI - Respiratory activation of the genioglossus muscle involves both non-NMDA and NMDA glutamate receptors at the hypoglossal motor nucleus in vivo. AB - Brainstem respiratory neurons innervate the hypoglossal motor nucleus which in turn transmits this respiratory drive signal to the genioglossus muscle of the tongue. The mechanism of this transmission is important to help maintain an open airspace for effective breathing, and is thought to rely almost exclusively on non-N-methyl-d-aspartate (non-NMDA) glutamate receptor activation during respiration. However those studies were performed in slices of medulla from neonatal animals in vitro which may have led to an underestimation of the contribution of NMDA glutamate receptors that may normally operate in intact preparations. The current study tests the hypothesis that both NMDA and non-NMDA receptors contribute to respiratory drive transmission at the hypoglossal motor nucleus in vivo. Experiments were performed in urethane-anesthetized and tracheotomized adult Wistar rats in which vagus nerves were either intact or sectioned. In the presence of augmented genioglossus activity produced by vagotomy, microdialysis perfusion of either an NMDA receptor antagonist (D-2 amino-5-phosphonovaleric acid, 0.001-10 mM) or a non-NMDA receptor antagonist (6 cyano-7-nitroquinoxaline-2, 3-dione disodium salt, 0.001-1 mM) to the hypoglossal motor nucleus reduced respiratory-related genioglossus activity in a dose dependent manner (P < 0.001) indicating that both NMDA and non-NMDA glutamate receptors are necessary for transmission of the respiratory drive signal to genioglossus muscle in vivo. Similar effects were observed in the vagus nerve intact rats. Further experiments demonstrated that each delivered antagonist had effects that were specific to its respective receptor. Regression analysis also revealed that the activity of both NMDA and non-NMDA receptors at the hypoglossal motor nucleus is related to levels of the prevailing respiratory drive. These results show that both NMDA and non-NMDA glutamate receptors at the hypoglossal motor nucleus are involved in transmission of the respiratory drive signal to genioglossus muscle in vivo. PMID- 16476524 TI - Genetic interdependence of adenosine and dopamine receptors: evidence from receptor knockout mice. AB - Dopamine and adenosine receptors are known to share a considerable overlap in their regional distribution, being especially rich in the basal ganglia. Dopamine and adenosine receptors have been demonstrated to exhibit a parallel distribution on certain neuronal populations, and even when not directly co-localized, relationships (both antagonistic and synergistic) have been described. This study was designed to investigate dopaminergic and purinergic systems in mice with ablations of individual dopamine or adenosine receptors. In situ hybridization histochemistry and autoradiography was used to examine the level of mRNA and protein expression of specific receptors and transporters in dopaminergic pathways. Expression of the mRNA encoding the dopamine D2 receptor was elevated in the caudate putamen of D1, D3 and A2A receptor knockout mice; this was mirrored by an increase in D2 receptor protein in D1 and D3 receptor knockout mice, but not in A2A knockout mice. Dopamine D1 receptor binding was decreased in the caudate putamen, nucleus accumbens, olfactory tubercle and ventral pallidum of D2 receptor knockout mice. In substantia nigra pars compacta, dopamine transporter mRNA expression was dramatically decreased in D3 receptor knockout mice, but elevated in A2A receptor knockout mice. All dopamine receptor knockout mice examined exhibited increased A2A receptor binding in the caudate putamen, nucleus accumbens and olfactory tubercle. These data are consistent with the existence of functional interactions between dopaminergic and purinergic systems in these reward and motor-related brain regions. PMID- 16476526 TI - Mitochondrial molecular clocks and the origin of the major Otocephalan clades (Pisces: Teleostei): A new insight. AB - The Otocephala, a clade including ostariophysan and clupeomorph teleosts, represents about a quarter of total fish species diversity, with about 1000 genera and more than 7000 species. A series of recent papers have defended that the origin of this clade and of its major groups may be significantly older than the oldest fossils of each of these groups suggest. Some of these recent papers explicitly defend a Pangean origin for some otocephalan groups such as the Siluriformes or Cypriniformes. To know whether or not the otocephalans as a whole, and particularly the mainly freshwater, cosmopolitan otophysans could have originated before the splitting of the Pangean supercontinent is of extreme importance, since otophysan fishes are among the most useful animal groups for the determination of historical continental relationships. In the present work we examined divergence times for each major otocephalan group by an analysis of complete mtDNA sequences, in order to investigate if these divergence times support the hypotheses advanced in recent studies. The complete mtDNA sequences of nine representative non-otocephalan fish species and of twenty-one representative otocephalan species was compared. The present study is thus, among the studies dealing with molecular divergence times of teleosts, the one in which a greater number of otocephalan species are included. The divergence times obtained support that the major otocephalan groups had a much older origin than the oldest fossil records available for these groups suggest. The origin of the Otocephala is estimated as having occurred about 282 Mya, with the origin of the Otophysi being estimated at about 251 Mya. PMID- 16476525 TI - Cancer prevention trials and primary care physicians: factors associated with recommending trial enrollment. AB - BACKGROUND: To explore the willingness of primary care providers (PCPs) to encourage enrollment of patients into cancer prevention trials. METHODS: A self administered survey was mailed to a random sample of PCPs in three geographic regions. Physicians were asked questions about their knowledge and attitudes towards cancer prevention trials. We presented a clinical vignette of a woman at high risk for breast cancer and asked if they would encourage her enrollment into a breast cancer chemoprevention trial (yes/no). Each survey included one of 16 possible clinical vignettes where patient characteristics (age, race socioeconomic status, physical mobility and co-morbidity) varied dichotomously. Bivariate analyses and logistic models were used to examine the independent effects of patient and physician characteristics on physician decisions. RESULTS: Two hundred and sixty-six surveys (50% response) were analyzed. The mean age of respondents was 48; 54% were White, 35% Asian and 5% Black. By design physicians were evenly distributed by gender, specialty and geographic location. Overall, 53% would encourage enrollment into a breast cancer chemoprevention trial. Significant predictors of a recommendation to enroll were: geographic location in California or Georgia, younger vignette patient and anticipating an increase in patient trust after recommending enrollment. CONCLUSION: PCPs are less likely to encourage elderly patients to enroll into cancer chemoprevention trials. Decisions differ based on geographic location and perceived trust in the patient provider relationship. To achieve successful enrollment, trial investigators must continue to educate PCPs and ensure a strong PCP-patient relationship is maintained. PMID- 16476527 TI - Characterization of a heat-shock-inducible hsp70 gene of the green alga Volvox carteri. AB - The green alga Volvox carteri possesses several thousand cells, but just two cell types: large reproductive cells called gonidia, and small, biflagellate somatic cells. Gonidia are derived from large precursor cells that are created during embryogenesis by asymmetric cell divisions. The J domain protein GlsA (Gonidialess A) is required for these asymmetric divisions and is believed to function with an Hsp70 partner. As a first step toward identifying this partner, we cloned and characterized V. carteri hsp70A, which is orthologous to HSP70A of the related alga Chlamydomonas reinhardtii. Like HSP70A, V. carteri hsp70A contains multiple heat shock elements (HSEs) and is highly inducible by heat shock. Consistent with these properties, Volvox transformants that harbor a glsA antisense transgene that is driven by an hsp70A promoter fragment express Gls phenotypes that are temperature-dependent. hsp70A appears to be the only gene in the genome that encodes a cytoplasmic Hsp70, so we conclude that Hsp70A is clearly the best candidate to be the chaperone that participates with GlsA in asymmetric cell division. PMID- 16476528 TI - mNSC1 shows no evidence of protein-coding capacity. AB - The Mus musculus non-selective cation channel gene mNSC1 was used as a classical example of a gene derived from transposable elements. To study the evolution of mNSC1 in M. musculus we sequenced this locus in M. musculus, M. hortulanus, M. spretus, M. caroli, and M. pahari. We found that the previously published 1,275 bp coding region was not present in any of these species. We identified a second possible coding region that was present only in M. musculus. However, RT-PCR experiments did not confirm the expression of the second reading frame. Our findings suggest that mNSC1 lacks protein-coding capacity and highlight the need for comparative validation of all TE-containing genes. PMID- 16476529 TI - Assessment of reproductive status in male echidnas. AB - This study reports the development and application of techniques to assess the reproductive status of male echidnas. The pattern of testosterone secretion over a 24-h period in five echidnas was documented. Testosterone secretion after injection i.m. of either 1000 IU hCG (n=4) or 4 microg GnRH agonist (n=6) was determined to establish whether this could be used as a practical index of the prevailing steroidogenic capacity of the testes. hCG (1000 IU) was also used to assess seasonal changes in testosterone secretion in six echidnas over a 13-month period. Seasonal changes in testicular volume were examined by transabdominal ultrasonography. Electroejaculation was attempted to monitor seasonal changes in sperm production, which was also determined by spermatorrhea. There was no apparent diurnal pattern of testosterone secretion in echidnas and circulating concentrations of testosterone remained relatively low (maximum 1.2 ng/mL) and stable over 24h. Injection of hCG resulted in an increase (P<0.01; n=4) in testosterone concentration with a peak (2.9+/-0.3 ng/mL) approximately 4h after injection. GnRH also induced an increase (P<0.01; n=6) in circulating testosterone that was apparent after 1h (2.6+/-0.3 ng/mL) and concentrations remained elevated (3.4+/-0.3 ng/mL) for up to 8h after injection. Seasonal changes in testosterone secretion determined after injection of hCG, increased (P=0.03; n=6) from late-autumn, peaked in late-winter, and decreased by early spring. Testicular volume followed a similar seasonal pattern (P<0.01; n=6) with an increase from late-autumn, peak in winter and a decline in mid-spring. There was no seasonal change in live weight. Electroejaculation was attempted throughout two breeding seasons but no semen was obtained. Spermatorrhoea in the echidna was described for the first time and was subsequently used to assess seasonal sperm production. Spermatozoa were found in the urine from June to September. This study has demonstrated that exogenous hormones can be used to obtain an index of the prevailing steroidogenic capacity of the testes in echidnas, which is not apparent with repetitive non-stimulated samples over 24 h. The assessment of testosterone secretion after injection of trophic hormones provides a valuable and practical procedure for the assessment of reproductive status. Testicular ultrasonography and spermatorrhea are useful in assessing reproductive status and in this study were successfully used to determine seasonal reproduction in captive echidnas. PMID- 16476530 TI - Comparative assessment of two indices of drug induced permeability changes in the perfused rat intestine. AB - In the present study, two indices of acute intestinal permeability changes were investigated as measurements of drug induced intestinal damage. The first method was based on 14C-polyethylene glycol (PEG) 4000 permeability assessment and the second was based on histological evaluation of the intestine. The test compounds were ibuprofen, ketoprofen and naproxen and the alanine, glycine and phenylalanine amide derivatives of ibuprofen. Perfusion studies were carried out using a rat model. Post-perfusion, the gut was fixed and tissue changes were assessed and scored. Ibuprofen, ketoprofen and naproxen altered the barrier properties of the intestine to PEG 4000 with significantly higher scores (p<0.05) for gastrointestinal toxicity relative to blank buffer. For ketoprofen, PEG 4000 permeability and intestinal damage scores increased with increasing ketoprofen concentration. Ibuprofen amide derivatives did not induce significant histological damage or PEG 4000 permeability when compared with ibuprofen. A correlation coefficient of 0.91 is obtained when intestinal damage scores are plotted against PEG 4000 permeability for all compounds. Both indices are proposed as rapid and useful measures of drug induced acute intestinal damage. PMID- 16476531 TI - Synthesis and evaluation of the mucoadhesivity of a CD-chitosan derivative. AB - Combining mucoadhesive characteristics of a biodegradable polymer such as chitosan with the potential to enhance drug release by increasing the solubility of poorly water-soluble drugs has great potential for pharmaceutical technology and drug delivery design. Polymeric delivery systems have been extensively researched in an attempt to achieve modified drug release. Cyclodextrins (CD) offer an alternative approach. These cyclic oligosaccharides have the ability to form non-covalent complexes with a number of drugs altering their physicochemical properties. In the continuing challenge to improve the properties of delivery systems, this paper focuses on the modification of chitosan by introducing beta cyclodextrin and to test the mucoadhesive strength and inclusion properties of this synthesised cyclodextrin-polymer. beta-Cyclodextrin was successfully grafted onto a chitosan chain polymer with a cyclodextrin grafting yield of 7% and a CD chitosan yield of 85%. Although the complexation of (+)-catechin by the grafted beta-CD was found to be about five times weaker than that by the beta-CD monoaldehyde and natural beta-CD, the inclusion properties of the chitosan-CD remain promising. The mucoadhesive properties of chitosan-CD were compared to that of pectin (reference) and the parent chitosan with the use of a tensile separation test. The chitosan-CD showed mucoadhesive strengths of 12% stronger than pectin, but 13.5% weaker than the parent chitosan. The synthesised chitosan CD-polymer exhibits characteristics of a possible mucoadhesive drug delivery system with some inclusion properties from beta-cyclodextrin. PMID- 16476532 TI - Three-dimensional regular arrangement of the annular ligament of the rat stapediovestibular joint. AB - The stapes footplate articulates with the vestibular window through the annular ligament. This articulation is known as the stapediovestibular joint (SVJ). We investigated the ultrastructure of adult rat SVJ and report here on the characteristic ultrastructure of the corresponding annular ligament. Transmission electron microscopy showed that this annular ligament comprises thick ligament fibers consisting of a peripheral mantle of microfibrils and an electron-lucent central amorphous substance that is regularly arranged in a linear fashion, forming laminated structures parallel to the horizontal plane of the SVJ. Scanning electron microscopy revealed that transverse microfibrils cross the thick ligament fibers, showing a lattice-like structure. The annular ligament was vividly stained with elastica van Gieson's stain and the Verhoeff's iron hematoxylin method. Staining of the electron-lucent central amorphous substance of the thick ligament fibers by the tannate-metal salt method revealed an intense electron density. These results indicate that the annular ligament of the SVJ is mainly composed of mature elastic fibers. PMID- 16476533 TI - Lymph node staging with dual-modality PET/CT: enhancing the diagnostic accuracy in oncology. AB - Lymph node staging according to the TNM criteria is an essential part of tumor evaluation. Several morphological and functional imaging procedures are used complementarily in this setting. Dual-modality PET/CT scanners are able to provide anatomical and functional data sets in a single session with accurate image co-registration. Comparative studies between morphological imaging procedures, such as MRI and CT, with co-registered PET/CT demonstrated significantly better lymph node staging with PET/CT than with anatomical procedures alone, regardless of the staged body compartment (head and neck, thorax or abdominal area). Based on more accurate staging results, PET/CT was able to alter the patients' therapy in a significant number of studies. Functional imaging with FDG-PET ([(18)F]-2-fluoro-2-desoxy-D-glucose-positron emission tomography) demonstrated outstanding results in lymph node staging of different tumor diseases. By adding anatomical information to PET, PET/CT outperforms PET alone when assessing the TNM-stage of different malignant diseases. This paper provides an overview concerning the performance of PET/CT in staging lymph nodes for malignant spread and points out benefits and limitations of this new imaging modality. PMID- 16476534 TI - The superior vena cava syndrome caused by malignant disease. Imaging with multi detector row CT. AB - OBJECTIVE: The superior vena cava (SVC) obstruction by malignant diseases is either by direct invasion and compression or by tumour thrombus of the SVC. Whatever is its cause, obstruction of the SVC causes elevated pressure in the veins draining into the SVC and increased or reversed blood flow through collateral vessels. Severity of the syndrome depends on the collateral vascular system development. Therefore, imaging of the collateral veins with variable location and connection is important in determining the extension and management of the disease. Our aims are to describe collateral vessels of the superior vena cava syndrome (SVCS) related with the malignant diseases and to assess the ability of multi-detector row CT with multiplanar and 3D volume rendering techniques in determining and describing collateral circulations. MATERIALS AND METHODS: We present CT angiography findings of seven patients with small cell carcinoma of the lung (n = 2), squamous cell carcinoma of the lung (n = 3), Hodgkin disease of the thorax (n = 1), and squamous cell carcinoma of the oesophagus (n = 1). The patients received contrast-enhanced CT scans of the chest and abdomen on a multi-detector row CT during breath holding at suspended inspiration. RESULTS: CT images revealed the cause and level of the SVC obstruction in all patients with axial and multiplanar reconstructed images. The SVC showed total obstruction in five patients and partial obstruction in two patients. The most common experienced collateral vessels were azygos vein (6), intercostal veins (6), mediastinal veins (6), paravertebral veins (5), hemiazygos vein (5), thoracoepigastric vein (5), internal mammary vein (5), thoracoacromioclavicular venous plexus (5), and anterior chest wall veins (5). While one case showed the portal-systemic shunt, V. cordis media and sinus coronarius with phrenic veins were enlarged in two cases, and the left adrenal vein was enlarged in a patient. In one case, the azygos vein with reversed blood flow was drained into both inferior vena cava and hemiazygos vein with the left renal vein. CONCLUSION: Multi-detector row CT with multiplanar and 3D imaging is an effective tool in evaluation of the SVCS and has a greater advantage than the other imaging techniques. 3D volume rendering is a useful technique in determining and describing collateral circulations in addition to the primary disease process. PMID- 16476535 TI - Classification and segmentation of intracardiac masses in cardiac tumor echocardiograms. AB - This paper describes an automatic method for classification and segmentation of different intracardiac masses in tumor echocardiograms. Identification of mass type is highly desirable, since to different treatment options for cardiac tumors (surgical resection) and thrombi (effective anticoagulant treatment) are possible. Correct diagnosis of the character of intracardiac mass in a living patient is a true challenge for a cardiologist; therefore, an objective image analysis method may be useful in heart diseases diagnosis. Image texture analysis is used to distinguish various types of masses. The presented methods assume that image texture encodes important histological features of masses and, therefore, texture numerical parameters enable the discrimination and segmentation of a mass. The recently developed technique based on the network of synchronized oscillators is proposed for the image segmentation. This technique is based on a 'temporary correlation' theory, which attempts to explain scene recognition as it would be performed by a human brain. This theory assumes that different groups of neural cells encode different properties of homogeneous image regions (e.g. shape, color, texture). Monitoring of temporal activity of cell groups leads to scene segmentation. A network of synchronized oscillators was successfully used for segmentation of Brodatz textures and medical textured images. The advantage of this network is its ability to detect texture boundaries. It can be also manufactured as a VLSI chip, for a very fast image segmentation. The accuracy of locating of analyzed tissues in the image should be assessed to evaluate a segmentation technique. The new evaluation method based on measurement of physical textured test objects was proposed. Firstly, a series of object images was obtained by the use of different devices (scanner, digital camera and TV camera). Secondly, the images were segmented using oscillator network and feedforward artificial neural network. Thirdly, geometrical test object parameters were estimated and compared to its true values. The experiment was repeated also for ultrasound images, which represented rectangular cross-section of synthetic sponge submerged in water. In addition, classification and segmentation of selected benign tumor echocardiograms were performed. Oscillator network was used with network weights defined for both whole texture region and texture boundary detection for the tumor segmentation. The latter method provides much faster segmentation with the similar accuracy. The obtained segmentation results were discussed and compared to the artificial neural network classifier. Finally, it was demonstrated that the network of synchronized oscillators is a reliable tool for the segmentation of the selected intracardiac masses, since it gives a relatively accurate location of analyzed tissues. The advantage of the proposed method is its resistance to changes of the visual information in the analyzed image and to noise and artifacts, often present in echocardiograms. PMID- 16476536 TI - PAMPA--a drug absorption in vitro model 13. Chemical selectivity due to membrane hydrogen bonding: in combo comparisons of HDM-, DOPC-, and DS-PAMPA models. AB - This study compares the intrinsic permeability coefficients of 40 drug molecules, obtained by three popular variants of the PAMPA assay, based on: (a) n hexadecane, (b) 2% w/v dioleyoylphosphatidylcholine in n-dodecane, and (c) 20% w/v lecithin in n-dodecane, the HDM-, DOPC-, DS-PAMPA models, respectively. It was shown that PAMPA permeability values consistently rank in magnitude according to: DS>DOPC>HDM, with molecules like metoprolol showing 1000-fold greater permeability in DS than in HDM. Abraham descriptors were used to rationalize these observations. Water-solubilized polar molecules form very strong H-bonds with the solvent. Such molecules need to break these bonds in order to enter the pure alkane phase, which, in turn, offers no compensating H-bond solvation. Thus, more energy appears to be needed for a polar molecule to penetrate a pure alkane barrier, compared to a barrier possessing some H-bond interactions. The 20% phospholipid content of the DS-PAMPA lipid may be thought to ease the permeation process, by offering a compensating source of H-bonding within the membrane phase. PMID- 16476537 TI - Deficient alpha-1-antitrypsin phenotypes and persistent airflow limitation in severe asthma. AB - BACKGROUND: Persistent airflow limitation is common among patients with severe asthma, but its pathogenesis has not been fully clarified. Severe alpha-1 antitrypsin (AAT) deficiency is a risk factor of chronic airflow limitation and emphysema, and partially deficient phenotypes have been associated with an accelerated decline in lung function. We hypothesized that partial deficiency of AAT (non-PiM AAT phenotype) is a risk factor of persistent airflow limitation in asthma. METHODS: In 122 patients with severe asthma (86 females; age (median (range)): 44.0 yr (18-75)) postbronchodilator FEV1 and FEV1/VC were measured and the AAT phenotype was determined. Persistent airflow limitation was defined as postbronchodilator FEV1 or FEV1/VC < 75% pred. with TLC > 75% pred. RESULTS: Six patients (4.9%) had a non-PiM phenotype (1 MF, 3 MS, 1 MZ and 1 SZ). Of the 58 patients with persistent airflow limitation only 1 patient (1.7%) had a non-PiM phenotype vs. 7.8% among the patients without persistent airflow limitation (P = 0.21). Postbronchodilator FEV1/VC (% pred.) was higher in the non-PiM patients than in the PiM patients (P = 0.02), the other lung function parameters were not different. Linear regression analysis showed no association between AAT phenotype and FEV1% predicted (P = 0.26). CONCLUSIONS: AAT heterozygoty does not seem to be an important risk factor of persistent airflow limitation in patients with asthma. Although confirmation by longitudinal follow-up studies with larger sample sizes is needed, these results suggest that routine assessment of the AAT phenotype is not indicated in asthmatic patients even if they exhibit fixed airflow limitation. PMID- 16476538 TI - Transcatheter closure of a patent foramen ovale prior to a pneumonectomy to prevent platypnea syndrome. AB - An intracardiac shunt on an atrial level may be a cause of hypoxemia after lung surgery (platypnea-orthodeoxia syndrome). This syndrome may be successfully treated by percutaneous or surgical shunt closure. We present the case of preoperative transcatheter closure of a patent foramen ovale (PFO) for the prevention of possible platypnea after a forthcoming right pneumonectomy. PMID- 16476539 TI - 'Trivial tricuspid regurgitation' -- is the impact really trivial? PMID- 16476540 TI - Ossification of the intercostal muscle around the bronchial anastomosis does not jeopardize airway patency. PMID- 16476541 TI - Mediastinal bronchogenic cyst's recurrence treated with EBUS-FNA with a long-term follow-up. AB - Bronchogenic cysts are congenital abnormalities generally mediastinal and are frequently detected incidentally. We report a case of a symptomatic mediastinal cyst treated previously by video-assisted thoracoscopy (VATS) but complicated by pericystic adhesions. The subsequent incomplete excision led, after 8 months, to a cyst's recurrence that was accurately drained by endobronchial ultrasonography guided fine needle aspiration (EBUS-FNA), with no new regrowth after 18 months. We hypothesized that the support of a high-definition diagnostic tool (EBUS) improved the FNA ability to make a deep and complete aspiration of the cyst. The usefulness of FNA in bronchogenic cyst's treatment is underestimated. Our experience is an attempt to encourage the use of EBUS-FNA as a new therapeutic option in the management of bronchogenic cyst. PMID- 16476542 TI - The degree of oxidative stress is associated with major adverse effects after lung resection: a prospective study. AB - OBJECTIVE: This prospective randomized study was conducted in order to define the contribution of the generated oxygen and nitrogen reactive species on postlobectomy morbidity and mortality. PATIENTS AND METHODS: Between 2001 and 2003, 132 patients with non-small cell lung cancer (NSCLC) were prospectively studied. The patients were grouped according to one-lung ventilation (OLV) use or not and to the duration of lung's atelectasis. Group A included 50 patients with confirmed non-small cell lung cancer who were subjected to lobectomy without one lung ventilation. Group B included 30 patients subjected to 60 min OLV. Group C included 30 patients subjected to 90 min OLV. Group D included 22 patients subjected to 120 min OLV. Preoperative, intraoperative and postoperative strict blood sampling protocol was followed. Malondialdehyde (MDA) plasma levels were measured. The groups were statistically compared for the occurrence of postoperative complications. OLV (groups B-D) along with other clinical parameters were entered in multivariate analysis as risk factors for complication development. MEASUREMENTS AND RESULTS: Comparison of group A with groups B-D (OLV) documented significant increase (p<0.001) of MDA levels during lung reexpansion. The magnitude of oxidative stress was related to OLV duration (group D>group C>group B, all p<0.001). Univariate analysis disclosed a higher incidence of acute respiratory failure, cardiac arrhythmias and pulmonary hypertension in group D. Multivariate analysis revealed OLV as an independent risk factor for postoperative development of cardiac arrhythmias and pulmonary hypertension. CONCLUSION: Protracted (>1h) OLV should be considered a potential cause for cardiovascular complications through the generation of severe oxidative stress due to lung reexpansion. PMID- 16476543 TI - Early removal of chest drainage and outpatient program after videothoracoscopic lung biopsy. PMID- 16476544 TI - Giant ascending aorta aneurysm in an 82-year-old woman. PMID- 16476546 TI - Haemothorax with posterior chest wall flail segment due to traumatic spinal cord transection. PMID- 16476547 TI - Repair of persistent truncus arteriosus with interrupted aortic arch: what did we learn? PMID- 16476548 TI - Heart failure following left-sided pneumonectomy in a patient with known pectus excavatum -- successful treatment using the Ravitch procedure. AB - A rare case of a patient with progressive dyspnoea due to atrial compression between ascending and descending aorta is demonstrated. After neoadjuvant chemoradiation for a locally advanced nonsmall cell lung cancer stage IIIb, he had a left-sided pneumonectomy. The underlying problem for cardiac compression was the extreme mediastinal shift reinforced by a congenital pectus excavatum. Our treatment was a Ravitch procedure with fair result. PMID- 16476551 TI - Abdominal visceral hernia through a chest wall defect secondary to open-window thoracostomy. PMID- 16476552 TI - A novel technique using echocardiography to evaluate venous cannula performance perioperatively in CPB cardiac surgery. AB - OBJECTIVE: Transthoracic echocardiography (TTE) has been used clinically to disobstruct venous drainage cannula and to optimise placement of venous cannulae in the vena cava but it has never been used to evaluate performance capabilities. Also, little progress has been made in venous cannula design in order to optimise venous return to the heart lung machine. We designed a self-expandable Smartcanula (SC) and analysed its performance capability using echocardiography. METHODS: An epicardial echocardiography probe was placed over the SC or control cannula (CTRL) and a Doppler image was obtained. Mean (V(m)) and maximum (V(max)) velocities, flow and diameter were obtained. Also, pressure drop (DeltaP(CPB)) was obtained between the central venous pressure and inlet to venous reservoir. LDH and Free Hb were also compared in 30 patients. Comparison was made between the two groups using the student's t-test with statistical significance established when p<0.05. RESULTS: Age for the SC and CC groups were 61.6+/-17.6 years and 64.6+/-13.1 years, respectively. Weight was 70.3+/-11.6 kg and 72.8+/ 14.4 kg, respectively. BSA was 1.80+/-0.2 m(2) and 1.82+/-0.2 m(2), respectively. CPB times were 114+/-53 min and 108+/-44 min, respectively. Cross-clamp time was 59+/-15 min and 76+/-29 min, respectively (p=NS). Free-Hb was 568+/-142 U/l versus 549+/-271 U/l post-CPB for the SC and CC, respectively (p=NS). LDH was 335+/-73 mg/l versus 354+/-116 mg/l for the SC and CC, respectively (p=NS). V(m) was 89+/-10 cm/s (SC) versus 63+/-3 cm/s (CC), V(max) was 139+/-23 cm/s (SC) versus 93+/-11 cm/s (CC) (both p<0.01). DeltaP(CPB) was 30+/-10 mmHg (SC) versus 43+/-13 mmHg (CC) (p<0.05). A Bland-Altman test showed good agreement between the two devices used concerning flow rate calculations between CPB and TTE (bias 300 ml+/-700 ml standard deviation). CONCLUSIONS: This novel Smartcanula design, due to its self-expanding principle, provides superior flow characteristics compared to classic two stage venous cannula used for adult CPB surgery. No detrimental effects were observed concerning blood damage. Echocardiography was effective in analysing venous cannula performance and velocity patterns. PMID- 16476553 TI - Regulatory T cells in transplantation. AB - Our ability to harness tolerance mechanisms will have a major impact in organ transplantation. It should enable drug minimization, and eventually, the elimination of all immunosuppressive drugs. An improved understanding of the biology of regulatory T cells will make it possible to replace current induction regimens with those favouring the selective vaccination of T cells that prevent graft rejection. Once regulation is established, the continued supply of graft antigens should empower T cell regulation to become the dominant natural mechanism to prevent graft rejection. PMID- 16476554 TI - The effect of oral immuno-stimulation in juvenile carp (Cyprinus carpio L.). AB - The effect of a 2-week period of oral immuno-stimulation from the age of 2 or 6 weeks post-fertilisation (wpf; before and after reaching the ability to produce antibodies) onwards was investigated on various immune functions of the common carp, Cyprinus carpio. The immuno-stimulants Aeromonas salmonicida lipopolysaccharide, Yeast DNA (containing unmethylated CpG motifs) or high-M alginate (an extract of algae containing poly-mannuronic acid) were used. The effect of this treatment was studied on the kinetics of B cells in head kidney and peripheral blood leucocytes using flow cytometry, on the total plasma IgM level using ELISA, on cytokine and inducible nitric oxide synthase (iNOS) expression in the intestine, and acute phase protein expression in the liver, using real time quantitative PCR, and on exposure to Vibrio anguillarum. Oral administration of immuno-stimulants from 6 wpf resulted in decreased WCI12(+) (B) cell percentages in PBL (only after administration of LPS) and head kidney (all test groups), and a decreased total IgM level in plasma, suggesting that suppressive effects are strongly indicative of oral or juvenile tolerance. After administration from 2 wpf, the effects on WCI12(+) (B) cell percentages were less pronounced: the group fed with Yeast DNA showed higher percentages compared to the control group at 6 wpf, but lower percentages at 8 wpf. No changes were observed in the cytokine or iNOS expression levels in the intestine or acute phase protein expression in the liver. A challenge with V. anguillarum resulted in an initially higher cumulative mortality in the group fed with LPS, but lower mortality in the groups fed with Yeast DNA or high-M alginate compared to the control group, providing a provisional warning especially for the use of pathogen derived immuno-stimulants, such as A. salmonicida LPS, in larval and juvenile fish. PMID- 16476555 TI - A composition-based cartilage model for the assessment of compositional changes during cartilage damage and adaptation. AB - OBJECTIVE: The composition of articular cartilage changes with progression of osteoarthritis. Since compositional changes are associated with changes in the mechanical properties of the tissue, they are relevant for understanding how mechanical loading induces progression. The objective of this study is to present a computational model of articular cartilage which enables to study the interaction between composition and mechanics. METHODS: Our previously developed fibril-reinforced poroviscoelastic swelling model for articular cartilage was combined with our tissue composition-based model. In the combined model both the depth- and strain-dependencies of the permeability are governed by tissue composition. All local mechanical properties in the combined model are directly related to the local composition of the tissue, i.e., to the local amounts of proteoglycans and collagens and to tissue anisotropy. RESULTS: Solely based on the composition of the cartilage, we were able to predict the equilibrium and transient response of articular cartilage during confined compression, unconfined compression, indentation and two different 1D-swelling tests, simultaneously. CONCLUSION: Since both the static and the time-dependent mechanical properties have now become fully dependent on tissue composition, the model allows assessing the mechanical consequences of compositional changes seen during osteoarthritis without further assumptions. This is a major step forward in quantitative evaluations of osteoarthritis progression. PMID- 16476556 TI - Usefulness of a national hospital database to evaluate the burden of primary joint replacement for coxarthrosis and gonarthrosis in patients aged over 40 years. AB - OBJECTIVE: To evaluate the 2001 French burden of hospital primary joint replacement (PJR) for coxarthrosis and gonarthrosis. METHODS: Hospital surgical admissions for coxarthrosis and gonarthrosis in people aged over 40 years were selected from the French National Hospital Database. Of the 73,150 and 58,746 admissions for coxarthrosis and gonarthrosis, respectively, only 96 and 73% of them were analysed (exclusion of stays with no respect of coding guidelines). For each, we described the type of osteoarthritis, gender and age group distribution, incidence rate of PJR adjusted on age and gender, the type of joint replacement (total vs partial), the type of hospital (private vs hospital), the mean length of stay (LOS), the percentage of patients transferred to rehabilitation centre and the hospital costs. RESULTS: Whatever the type of osteoarthritis, PJR was mainly performed for primary osteoarthritis, in the 71-80 years' age group, in private hospital, with a total replacement procedure. The mean LOS were 13 and 12 days, and the transfers to a rehabilitation centre were 33 and 44%, for hip and knee, respectively. The incident rate of PJR increased significantly with age. It was higher in the 71-80 years' age group and decreased thereafter, whatever the gender and the type of osteoarthritis. The whole hospital costs were 591 and 411 millions of euros for hip and knee, respectively. CONCLUSION: The French National Hospital Database is a useful tool for assessing the burden of primary PJR for coxarthrosis and gonarthrosis. It might be used for international comparisons. PMID- 16476558 TI - Platelets in the onset of atherosclerosis. AB - Beyond their role in hemostasis and thrombosis platelets are critically involved in the onset of atherosclerosis. Platelets represent an important linkage between inflammation, and atherogenesis. Platelets interact with inflammatory cells including leukocytes and endothelium. These interactions lead to leukocyte recruitment towards the vascular wall, initiating extravasation of circulating mononuclear cells and foam cell generation. Inflammatory processes within the arterial wall result in development of atherosclerotic lesions and atheroprogression. Inhibition of platelet interaction with the arterial wall results in attenuation of atherosclerosis and may be a novel therapeutic strategy in treatment of high-risk patients. PMID- 16476557 TI - Ecto-nucleotidases of the CD39/NTPDase family modulate platelet activation and thrombus formation: Potential as therapeutic targets. AB - Extracellular nucleotide P2-receptor-mediated effects on platelets, leukocytes and endothelium are modulated by ecto-nucleotidases. These ecto-enzymes hydrolyze extracellular nucleotides to the respective nucleosides. The dominant ecto nucleotidase expressed by the endothelium, by monocytes and vascular smooth muscle cells is CD39/NTPDase1. Ecto-nucleotidase biochemical activity of CD39 is lost at sites of acute vascular injury, such as in ischemia reperfusion and immune graft rejection. CD39L(Like)1/NTPDase2, a related protein, is associated with the basolateral surface of endothelium, the adventitia of vessels and microvascular pericytes. CD39/NTPDase1 hydrolyzes both tri- and diphosphonucleosides and blocks platelet aggregation responses to ADP. In contrast, CD39L1/NTPDase2, a preferential nucleoside triphosphatase, activates platelets by preferentially converting ATP to ADP, the major agonist of platelet P2 receptors. Spatial and temporal expression of NTPDases in the vasculature appears to control platelet activation, thrombus size and stability by regulating phosphohydrolytic activity and consequent P2 receptor signaling. Constitutively circulating microparticles appear to be associated with functional NTPDases, and accumulation of these at sites of vascular injury might influence local thrombus formation and evolution. The phenotype of the cd39-null mouse is in keeping with disordered thromboregulation with heightened susceptibility to inflammatory vasculary reactions, increased permeability and high levels of tissue fibrin. Paradoxically, these mutant mice also exhibit a bleeding phenotype with differential platelet P2Y1 desensitization. Over-expression of CD39 at sites of vascular injury and inflammation by adenoviral vectors, by transgenesis or by the use of pharmacological modalities with soluble derivatives has been shown to have major potential in several animal models tested to date. Future clinical applications will involve the development of new therapeutic strategies to various inflammatory vascular diseases and in transplantation. PMID- 16476559 TI - Characterisation of the signalment, clinical and survival characteristics of 41 cats with mast cell neoplasia. AB - Mast cell tumours (MCTs) are relatively common tumours of cats, and are the second most common cutaneous tumours in cats in the USA. While the primary splenic form of the disease is far less common, it is usually associated with more severe clinical signs. Signalment, clinical and survival characteristics of mast cell neoplasia were characterised in 41 cats. The most common tumour location was cutaneous/subcutaneous head and trunk. Stage 1a was the most common tumour stage at first diagnosis (n=20), followed by stage 4 (both stage 4a and stage 4b; n=10). Of 22 cats that underwent excisional biopsy, mast cell neoplasia recurred in four cats during the study period. Three of the 41 cats presented with simultaneous cutaneous and either splenic or lymph node tumours. A comparison between cats with only cutaneous tumours (n=30) and those with tumours involving the spleen or lymph nodes (n=11) showed longer survival times for the cutaneous-only group (P=0.031). Twelve of the 41 cats died of mast cell neoplasia during the study period. When a subgroup of cats with only cutaneous tumours (no lymph node or visceral involvement) were divided according to whether there were multiple (five or more) tumours (n=6) or a single tumour (n=19), cats with single tumours survived longer than those with multiple tumours (P=0.001). Solitary cutaneous feline MCTs without spread to the lymph nodes usually manifest as benign disease with a relatively protracted course. However, multiple cutaneous tumours, recurrent tumours and primary splenic disease should receive a guarded prognosis due to the relatively short median survival times associated with these forms of the disease. PMID- 16476560 TI - Prevalence of antibodies against feline coronavirus and Chlamydophila felis in Swedish cats. AB - Serum samples from 214 Swedish cats with no signs of infectious disease were analysed for the presence of antibodies against Chlamydophila felis (Cp felis), while 209 of these were also analysed for feline coronavirus (FCoV) antibodies. The prevalence of antibodies against Cp felis was 11%, with no significant difference between purebred and mixed breed cats. The overall prevalence of antibodies against FCoV was 31%, significantly higher among pure breed cats (65%) than among mixed breed cats (17%). A high proportion of cats with antibodies against FCoV had relatively high antibody titres, and was therefore likely to be shedding FCoV in faeces. For Cp felis, the majority of seropositive animals had relatively low antibody titres, and the risk of these animals infecting others is not known. PMID- 16476561 TI - A combined immuno-informatics and structure-based modeling approach for prediction of T cell epitopes of secretory proteins of Mycobacterium tuberculosis. AB - The role of secretory proteins of Mycobacterium tuberculosis in pathogenesis and stimulation of specific host responses is well documented. They are also shown to activate different cell types, which subsequently present mycobacterial antigens to T cells. Therefore identification of T cell epitopes from this set of proteins may serve to define candidate antigens with vaccine potential. Fifty-two secretory proteins of M. tuberculosis H37Rv were analyzed computationally for the presence of HLA class I binding nonameric peptides. All possible overlapping nonameric peptide sequences from 52 secretory proteins were generated in silico and analyzed for their ability to bind to 33 alleles belonging to A, B and C loci of HLA class I. Fifteen percent of generated peptides are predicted to bind to HLA with halftime of dissociation T(1/2) >or=100 min and 73% of the peptides predicted to bind are mono-allelic in their binding. The structural basis for recognition of no-namers by different HLA molecules was studied employing structural modeling of HLA class I-peptide complexes and there exists a good correlation between structural analysis and binding prediction. Pathogen peptides that could behave as self- or partially self-peptides in the host were eliminated using a comparative study with the human proteome, thus reducing the number of peptides for analysis. The implications of the finding for vaccine development are discussed vis-a-vis the limitations of the use of subunit vaccine and DNA vaccine. PMID- 16476562 TI - Proinflammatory gene expression in mouse ST2 cell line in response to infection by Porphyromonas gingivalis. AB - Porphyromonas gingivalis is a predominant periodontal pathogen, whose infection causes inflammatory responses in periodontal tissue and alveolar bone resorption. Various virulence factors of this pathogen modulate host innate immune responses. It has been reported that gingipains degrade a wide variety of host cell proteins, and fimbriae are involved in bacterial adhesion to and invasion of host cells. In the present study, we profiled ST2 stromal cell gene expression following infection with the viable P. gingivalis strain ATCC33277 as well as with its gingipain- and fimbriae-deficient mutants, using microarray technology and quantitative real-time polymerase chain reaction. Using a mouse array of about 20,000 genes, we found that infection with the wild strain elicited a significant upregulation (greater than 2-fold) of expression of about 360 genes in ST2 cells, which included the chemokines CCL2, CCL5, and CXCL10, and other proinflammatory proteins such as interleukin-6 (IL-6) and matrix metalloproteinase-13 (MMP-13). Further, infection with the gingipain-deficient mutant elicited a reduced expression of the CXCL10, IL-6 and MMP-13 genes, suggesting that gingipains play an important role in inducing the expression of those genes following P. gingivalis infection. On the other hand, the pattern of global gene expression induced by the fimbriae-deficient mutant was similar to that by the wild strain. These results suggest that P. gingivalis infection induces gene expression of a wide variety of proinflammatory proteins in stromal cells/osteoblasts, and gingipains may be involved in inducing several of the proinflammatory factors. PMID- 16476563 TI - Induction of apoptosis by Legionella pneumophila in mammalian cells requires the mitochondrial pathway for caspase activation. AB - Legionella pneumophila, the agent of human Legionnaire's disease is a Gram negative, rod-shaped bacterium. During infection, the bacteria invade human cells and replicate intracellularly. L. pneumophila can induce apoptosis in human myeloid and epitheloid cells and this may contribute to the development of pathology and disease. However, the molecular mechanism of apoptosis induction is still uncertain. Here we investigate this process. Legionella efficiently induced apoptosis in myeloid cells, T cells and fibroblasts. Induction of apoptosis involved activation of the initiator caspase-9 and effector caspases. Caspase activity was required for cell death. Analysis of mutant cells showed that the death receptor pathway was not involved in Legionella-induced apoptosis. Surprisingly, caspase activity was found almost exclusively in cells that did not harbor bacteria. Infection with Legionella caused the activation of the pro apoptotic protein Bax and the release of cytochrome c. Mouse embryonic fibroblasts deficient for Bax and/or Bak were protected from Legionella-induced caspase activation. These results show a clear contribution of the mitochondrial pathway to Legionella-induced apoptosis. PMID- 16476564 TI - Paracoccin, a GlcNAc-binding lectin from Paracoccidioides brasiliensis, binds to laminin and induces TNF-alpha production by macrophages. AB - Paracoccidioides brasiliensis components interact with host cells and can influence the pathogenesis of paracoccidioidomycosis (PCM). Among the components released by P. brasiliensis, gp 43 and a heavily glycosylated antigen with MM>160 kDa are the most recognized by serum antibodies from patients with PCM. In order to isolate the high MM glycoconjugate, we carried out affinity chromatography of a crude exoantigen preparation on immobilized jacalin. The bound fraction (JBE, jacalin binding exoantigen) consisted of a major antigen of high MM and frequently of an additional 70-kDa minor protein. This protein, designated paracoccin, exhibited selective binding to immobilized GlcNAc, a property that was used for its purification. The structural data of paracoccin obtained by mass spectrometry of tryptic peptides did not match any known protein. Anti-paracoccin serum localized the lectin on the surface of P. brasiliensis yeasts, especially in the budding regions. Paracoccin was able to interact with laminin in a dose dependent manner. This interaction was inhibited by GlcNAc, followed by D-glucose and D-mannose, but not by D-galactose, N-acetyl-galactosamine or L-fucose. Interestingly, paracoccin induced both resident and elicited mouse peritoneal cavity macrophages to release high and persistent levels of TNF-alpha in vitro, a fact that was associated with high nitric oxide production in elicited cells. Because binding to laminin can favor yeast adhesion and invasion of host tissues, and overproduction of NO has been associated with suppression of cell immunity, paracoccin is suggested to play an important role in PCM pathogenesis. PMID- 16476565 TI - The role of histamine in the intracellular survival of Mycobacterium bovis BCG. AB - The course and outcome of infection with mycobacteria are determined by a complex interplay between the immune system of the host and the survival mechanisms developed by the bacilli. Histamine plays an important role in various processes, including cell division, metabolism, and apoptosis, and it modulates innate and adaptive immune responses. In the present study we investigated the intracellular survival of Mycobacterium bovis BCG in murine bone-marrow macrophages isolated from wild-type (WT) and histidine-decarboxylase knock-out [HDC (-/-)] mice. Mycobacterial titers were significantly higher in the HDC (-/-) macrophages as compared with the WT cells. M. bovis BCG growth in WT macrophages could be enhanced by pyrilamine and cimetidine. Exogenously added histamine decreased the intracellular counts of M. bovis BCG in HDC (-/-) macrophages. Infection of activated macrophages with M. bovis BCG elicited apoptosis, but there was no significant difference between the WT and the HDC (-/-) cells. These bacilli induced comparable levels of tumor necrosis factor-alpha production in the WT and the HDC (-/-) macrophages. M. bovis BCG stimulated interleukin-18 (IL-18) production in the macrophages from WT mice, but not in the HDC (-/-) cells. Exogenously added IL-18 decreased the titers of intracellular mycobacteria in HDC (-/-) cells. In conclusion, these data implicate histamine in the intracellular survival of M. bovis BCG. The cellular control mechanisms restricting the growth of M. bovis BCG are complex and involve H1 and H2 receptor-mediated events. Histamine might be an important mediator of M. bovis BCG-induced IL-18 production, which in turn contributes to immune protection. PMID- 16476566 TI - Comparative study of morphological and time-frequency ECG descriptors for heartbeat classification. AB - The prompt and adequate detection of abnormal cardiac conditions by computer assisted long-term monitoring systems depends greatly on the reliability of the implemented ECG automatic analysis technique, which has to discriminate between different types of heartbeats. In this paper, we present a comparative study of the heartbeat classification abilities of two techniques for extraction of characteristic heartbeat features from the ECG: (i) QRS pattern recognition method for computation of a large collection of morphological QRS descriptors; (ii) Matching Pursuits algorithm for calculation of expansion coefficients, which represent the time-frequency correlation of the heartbeats with extracted learning basic waveforms. The Kth nearest neighbour classification rule has been applied for assessment of the performances of the two ECG feature sets with the MIT-BIH arrhythmia database for QRS classification in five heartbeat types (normal beats, left and right bundle branch blocks, premature ventricular contractions and paced beats), as well as with five learning datasets-one general learning set (GLS, containing 424 heartbeats) and four local sets (GLS+about 0.5, 3, 6, 12 min from the beginning of the ECG recording). The achieved accuracies by the two methods are sufficiently high and do not show significant differences. Although the GLS was selected to comprise almost all types of appearing heartbeat waveforms in each file, the guaranteed accuracy (sensitivity between 90.7% and 99%, specificity between 95.5% and 99.9%) was reasonably improved when including patient-specific local learning set (sensitivity between 94.8% and 99.9%, specificity between 98.6% and 99.9%), with optimal size found to be about 3 min. The repeating waveforms, like normal beats, blocks, paced beats are better classified by the Matching Pursuits time-frequency descriptors, while the wide variety of bizarre premature ventricular contractions are better recognized by the morphological descriptors. PMID- 16476567 TI - Effect of cyclosporin A on the biliary excretion of cholephilic compounds in rats. AB - Cyclosporin A (CsA) is known to cause cholestasis. CsA is reported to competitively inhibit the transport of the substrates of the bile salt export pump (Bsep), multidrug resistance protein 2 (Mrp2) and P-glycoprotein (P-gp) in the canalicular membrane vesicles. However, the inhibitory effect of CsA on various substrates of the canalicular ATP-dependent transporters in vivo is unknown. Therefore, in the present study, the acute effect of CsA on the biliary excretion of the substrates of Bsep, Mrp2 and P-gp was examined under the same condition. Ten minutes after the intravenous administration of CsA (25mg/kg), the biliary excretion of various bile acids and organic anions and cations was studied. CsA decreased the biliary excretion of tracer amounts of taurocholate, leukotriene C(4), estradiol-17beta-glucuronide, pravastatin, vinblastine and erythromycin. In contrast, the biliary excretion of high doses of taurocholate and sulfobromophthalein was only slightly or not inhibited by CsA. In conclusion, CsA may competitively inhibit biliary excretion of substrates of Bsep, Mrp2 and P gp also in vivo, and CsA is considered to inhibit bile acid-dependent bile flow by the competitive inhibition of the canalicular transport of bile acids by Bsep. PMID- 16476568 TI - Acetate generation in rat liver mitochondria; acetyl-CoA hydrolase activity is demonstrated by 3-ketoacyl-CoA thiolase. AB - Acetate has been found as an endogenous metabolite of beta-oxidation of fatty acids in liver. In order to investigate the regulation of acetate generation in liver mitochondria, we attempted to purify a mitochondrial acetyl-CoA hydrolase in rat liver. This acetyl-CoA-hydrolyzing activity in isolated mitochondria was induced by the treatment of rats with di(2-ehtylhexyl)phthalate (DEHP), a peroxisome proliferator which induces expression of several peroxisomal and mitochondrial enzymes involved in beta-oxidation of fatty acids. The purified enzyme was 43-kDa in molecular mass by SDS/PAGE. Internal amino acid sequencing of this enzyme revealed that it was identical with mitochondrial 3-ketoacyl-CoA thiolase, suggesting that this enzyme has two kinds of activities, 3-ketoacyl-CoA thiolase and acetyl-CoA hydrolase activities. Kinetic studies clearly indicated that this enzyme had the both activities and each activity was inhibited by the substrates of the other activity, that is, 3-ketoacyl-CoA thiolase activity was inhibited by acetyl-CoA, on the other hand, acetyl-CoA hydrolase activity was inhibited by acetoacetyl-CoA in a competitive manner. These findings suggested that acetate generation in liver mitochondria is a side reaction of this known enzyme, 3-ketoacyl-CoA thiolase, and this enzyme may regulate its activities depending on each substrate level. PMID- 16476569 TI - A network of networks: quorum-sensing gene regulation in Pseudomonas aeruginosa. AB - The opportunistic pathogen Pseudomonas aeruginosa possesses two N-acyl-homoserine lactone quorum-sensing systems that regulate large, overlapping sets of genes. Recent global transcriptome and proteome analyses provided a wealth of information about the identity of the regulated genes, N-acyl-homoserine lactone signal specificity, timing of induction, and environmental effects on gene expression. Quorum-sensing gene expression in P. aeruginosa is also embedded in a highly interconnected network of other regulatory systems with a high potential for integrating and responding to multiple environmental signals. Such epigenetic complexity may constitute the basis for the exceptional adaptability of P. aeruginosa to diverse environments. PMID- 16476571 TI - Correlation of T-lymphocyte subpopulations with immunological markers in HIV-1 infected Indian patients. AB - Progressive HIV disease is characterized by CD4 T cell decline and activation of the immune system. We aimed to study the quantitative alterations in the naive (CD45RA+CD62L+), memory/effector (CD45RO+) and activated (HLA-DR+CD38+) T lymphocyte subpopulations in antiretroviral treatment naive, HIV-1 infected Indian patients by three-color multi-parametric flow cytometry. The association of different CD4+ and CD8+ T cell subsets with the immunological markers- CD4+ and CD8+ T cell percentages was examined by calculating the partial correlation coefficients. We also observed significant differences in the expression of different CD4+ and CD8+ T-cell subsets among the two groups of patients formed using the median CD4+ T cell percentage value (15%) of the study population. The correlations of different CD4+ and CD8+ T cell subsets reflected the quantitative alterations in the T-lymphocyte subpopulations and activation of the immune system during HIV-infection. The study outcome also emphasizes the significance of the CD38+CD8+ T-lymphocyte subset as a prognostic marker for HIV management and ART monitoring in resource-limited settings of developing countries like India. PMID- 16476570 TI - The effect of a multidimensional exercise programme on symptoms and side-effects in cancer patients undergoing chemotherapy--the use of semi-structured diaries. AB - The aim of this study was to evaluate the effects of a 6-week intervention with structured physical activity, relaxation, body-awareness techniques and massage on the symptoms/side-effects of cancer patients undergoing chemotherapy. The study was prospective and exploratory, and 54 patients completed assessments for all 6 weeks of the intervention. In order to obtain a continuous record of side effects, a diary was developed for the patients' use throughout the intervention. The patients scored their symptoms/side-effects on a scale from 0 to 4, using the Common Toxicity Criteria and reported these scores in questionnaires. Twelve possible symptoms/side-effects were registered daily: lack of appetite, nausea, vomiting, diarrhea, paraesthesia, constipation, physical fatigue, mental fatigue, treatment-related fatigue, muscle pain, arthralgia and other pain. During the intervention a decrease in the scoring for 10 out of the 12 side-effects was found. Statistical significance was observed in the pain score (P=0.046) and the arbitrary-derived sum of the scores for symptoms and side-effects (P=0.036) respectively. Patients with evidence of disease (n=26) had significantly higher levels of symptoms/side-effects than patients with no evidence of disease (n=28) (P=0.027). The results indicate that a six weeks multidimensional exercise intervention undertaken by cancer patients with or without residual disease while undergoing chemotherapy can lead to a reduction in treatment-related symptoms. PMID- 16476572 TI - Brain natriuretic peptide plasma level is a reliable indicator of advanced diastolic dysfunction in patients with chronic heart failure. AB - The aim of the study was to determine the value of brain natriuretic peptide for the identification of diastolic dysfunction status in congestive heart failure. We studied 204 patients with stable heart failure. Brain natriuretic peptide plasma levels were correlated with echocardiographic parameters of diastolic dysfunction. Diastolic dysfunction was classified as mild (abnormal echocardiographic relaxation pattern) and severe (pseudo-normal or restrictive pattern). A significant correlation between brain natriuretic peptide levels and the other parameters was detected. Brain natriuretic peptide dosage, then, seems to be a reliable tool for the assessment of diastolic dysfunction status in patients with congestive heart failure. PMID- 16476573 TI - Novel gut afferents: Intrinsic afferent neurons and intestinofugal neurons. AB - Information about the conditions of all tissues in the body is conveyed to the central nervous system through afferent neurons. Uniquely amongst peripheral organs, the intestine has numerous additional afferent neurons, intrinsic primary afferent neurons that have their cell bodies and processes in the enteric plexuses and do not project to the central nervous system. They detect conditions within the gut and convey that information to intrinsic reflex pathways that are also entirely contained inside the gut wall. Intrinsic primary afferent neurons respond both to the presence of material in the gut lumen and to distension of the gut wall and initiate reflex changes in contractile activity, fluid transport across the mucosa and local blood flow. They also function as nociceptors that initiate tissue-protective propulsive and secretory reflexes to rid the gut of pathogens. The regulation of excitability of intrinsic primary afferent neurons is through multiple ion channels and ion channel regulators, and their excitability is critical to setting the strength of enteric reflexes. The intestine also provides afferent signals to sympathetic pre-vertebral ganglia. The signals are conveyed from the gut by intestinofugal neurons that have their cell bodies within enteric ganglia and form synapses in the sympathetic ganglia. Intestinofugal neurons form parts of the afferent limbs of entero-enteric inhibitory reflexes. Because the unusual afferent neurons of the small intestine and colon make their synaptic connections outside the central nervous system, the neurons and the reflex centres that they affect are potential targets for non central penetrant therapeutic compounds. PMID- 16476574 TI - Sensitization and conditioning as contributors to gastrointestinal vulnerability. AB - Electric shocks are stressful and if signalled can result in Pavlovian conditioning of the stress response. Stress arising from such shocks or conditioned psychological "threat" influences vulnerability to gastrointestinal disorders. Reviewed are our studies with rats showing that unconditioned stress experiences sensitize the glandular portion of the stomach to later restraint-in water induced erosions, as an animal model of ulcer disease. These stress effects are not attributable to corticoids but may be opioid/endorphin dependent. The unconditioned stress-induced sensitization is reduced by allowing the rat either control over or prediction of the shocks even though the direct experience with shocks is identical. Elicitation of the conditioned stress response by a signal during the ulcer induction or even shortly afterwards increases gastric vulnerability to erosions. We are now finding parallel unconditioned stress effects on colonic erosions and increases in intestinal permeability induced by dextran sulphate sodium, as an animal model of inflammatory bowel disease. We conclude that psychological context of past trauma and/or current threat increases vulnerability to gastrointestinal disorders. PMID- 16476575 TI - sHSPs under temperature and pressure: the opposite behaviour of lens alpha crystallins and yeast HSP26. AB - Small angle X-ray scattering was used to follow the temperature and pressure induced structural transitions of polydisperse native calf lens alpha-crystallins and recombinant human alphaB-crystallins and of monodisperse yeast HSP26. The alpha-crystallins were known to increase in size with increasing temperature, whereas HSP26 partially dissociates into dimers. SAXS intensity curves demonstrated that the average 40-mer calf alpha-crystallin converted into 80-mer in a narrow temperature range, from 60 to 69 degrees C, whereas the average 30 mer alphaB-crystallin was continuously transformed into 60-mer at lower temperature, from 40 to 60 degrees C. These temperature-induced transitions were irreversible. Similar transitions, yet reversible, could be induced with pressure in the 100 to 300 MPa pressure range. Moreover, temperature and pressure could be combined to lower the transition temperatures. On the other hand, SAXS curves recorded during pressure scans from 0.1 to 200 MPa with monodisperse 24-mer HSP26 revealed dissociation of the 24-mer into dimers. This dissociation was complete and reversible. Whatever the sHSP, a decrease of partial specific volume was found to be associated with the pressure induced quaternary structure transitions, in agreement with the hypothesis that such transitions represent a first step on the protein denaturation pathway. PMID- 16476577 TI - The loss of local HGF, an endogenous gastrotrophic factor, leads to mucosal injuries in the stomach of mice. AB - The stomach is constantly exposed to mechanical and chemical stresses. Under persistent damages, epithelial cell proliferation is required to maintain mucosal integrity. Nevertheless, which ligand system(s) is physiologically involved in gastric defense remains unclear. Herein, we provide evidence that HGF is a key "natural ligand" to reverse gastric injury. The injection of cisplatin in mice led to the loss of HGF in the gastric interstitium, associated with the decrease in proliferating epithelium and the progression of mucotitis. When c-Met tyrosine phosphorylation was abolished by anti-HGF IgG, mucosal cell proliferation became faint, leading to delayed recovery from mucotitis, and vice versa in cases of HGF supplementation. Our findings indicate that: (1) HGF/c-Met signal on mucosa is needed to restore gastric injuries; and (2) the loss of local HGF leads to manifestation of gastric lesions. This study provides a rationale that explains why HGF supplement is useful for reversing gastric diseases. PMID- 16476576 TI - Defective chondrocyte proliferation and differentiation in osteochondromas of MHE patients. AB - Multiple hereditary exostoses (MHE) is an autosomal dominant skeletal disorder caused by mutations in one of the two EXT genes and characterized by multiple osteochondromas that generally arise near the ends of growing long bones. Defective endochondral ossification is likely to be involved in the formation of osteochondromas. In order to investigate potential changes in chondrocyte proliferation and/or differentiation during this process, osteochondroma samples from MHE patients were obtained and used for genetic, morphological, immunohistological, and in situ hybridization studies. The expression patterns of IHH (Indian hedgehog) and FGFR3 (Fibroblast Growth Factor Receptor 3) were similar with transcripts expressed throughout osteochondromas. Expression of PTHR1 (Parathyroid Hormone Receptor 1) transcripts was restricted to a narrow zone of prehypertrophic chondrocytes. Numerous cells forming osteochondromas although resembling prehypertrophic chondrocytes, stained positively with an anti proliferating cell nuclear antigen (PCNA) antibody. In addition, ectopic expression of collagen type I and abnormal presence of osteocalcin (OC), osteopontin (OP), and bone sialoprotein (BSP) were observed in the cartilaginous osteochondromas. These data indicate that most chondrocytes involved in the growth of osteochondromas can proliferate, and that some of them exhibit bone forming cell characteristics. We conclude that in MHE, defective heparan sulfate biosynthesis caused by EXT mutations maintains the proliferative capacity of chondrocytes and promotes phenotypic modification to bone-forming cells. PMID- 16476578 TI - The corticosteroid hormone induced factor: a new modulator of KCNQ1 channels? AB - The corticosteroid hormone induced factor (CHIF) is a member of the one transmembrane segment protein family named FXYD, which also counts phospholemman and the Na,K-pump gamma-subunit. Originally it was suggested that CHIF could induce the expression of the I(Ks) current when expressed in Xenopus laevis oocytes, but recently CHIF has attracted attention as a modulatory subunit of the Na,K-pump. In renal and intestinal epithelia, the expression of CHIF is dramatically up-regulated in response to aldosterone stimulation, and regulation of epithelial ion channels by CHIF is an attractive hypothesis. To study a potential regulatory effect of the CHIF subunit on KCNQ1 channels, co-expression experiments were performed in Xenopus laevis oocytes and mammalian CHO-K1 cells. Electrophysiological characterization was obtained by two-electrode voltage-clamp and patch-clamp, respectively. In both expression systems, we find that CHIF drastically modulates the KCNQ1 current; in the presence of CHIF, the KCNQ1 channels open at all membrane potentials. Thereby, CHIF is the first accessory subunit shown to be capable of modulating both the Na,K-pump and an ion channel. To find a possible physiological function of the constitutively open KCNQ1/CHIF complex, the precise localization of KCNQ1 and CHIF in distal colon and kidney from control and salt-depleted rats was determined by confocal microscopy. However, in these tissues, we did not detect an obvious overlap in expression between KCNQ1 and CHIF. In conclusion, the hormone-regulated subunit CHIF modulates the voltage sensitivity of the KCNQ channels, but so far evidence for an actual co-localization of CHIF and KCNQ1 channels in native tissue is lacking. PMID- 16476579 TI - Impaired pain sensation in mice lacking Aquaporin-1 water channels. AB - Aquaporin-1 (AQP1), a membrane water channel, is expressed in choroid plexus where it contributes to cerebrospinal fluid production. Here, we show that AQP1 is also expressed in the dorsal horn of the spinal cord and the trigeminal nucleus caudalis, regions that process pain information. Within the dorsal root and trigeminal sensory ganglia, AQP1 is concentrated in small diameter cell bodies, most of which give rise to unmyelinated C-fibers. To study the role of AQP1 in pain signaling, we compared acute pain responses in wild-type mice and in mice lacking AQP1. AQP1(-/-) mice had reduced responsiveness to thermal and capsaicin chemical stimuli, but not to mechanical stimuli or formalin. These results provide evidence for AQP1 expression in nociceptive neurons and suggest that AQP1 may play a role in pain signal transduction. PMID- 16476580 TI - Interaction of Pin1 with Nek6 and characterization of their expression correlation in Chinese hepatocellular carcinoma patients. AB - The peptidyl-prolyl isomerase Pin1 is prevalently overexpressed in human cancers and is regarded as a new diagnostic and therapeutic target. Pin1 interacts with several proteins involved in cell cycle events in a phosphorylation-dependent manner. Among them, NIMA (never in mitosis, gene A) was first identified to interact with Pin1. In this report, we found that Pin1 could interact with Nek6, one of the human NIMA-related kinases (Neks). This interaction was confirmed by GST pull-down assay, which was further confirmed by immunoprecipitation experiments, as well as immunofluorescence colocalization. We further studied Pin1 and Nek6 mRNA level in 40 pairs of hepatocellular carcinoma cases, finding significant correlations between Nek6 and Pin1 mRNA expression levels in these samples. PMID- 16476581 TI - Screening for beta-poly(L-malate) binding proteins by affinity chromatography. AB - Poly(beta-L-malic acid) is a cell type-specific polymer of myxomycetes (true slime molds) with the physiological role to organize mobility of certain proteins over the giant multinucleated plasmodia. We have developed an affinity chromatography employing 1,6-diamino-n-hexane-Sepharose-coupled poly(malic acid) to identify such proteins in cellular extracts of Physarum polycephalum. Molecular masses were measured by SDS-PAGE and non-denaturing PAGE after silver staining and/or Western blotting. Protein complexes/subunits were detected by 2 dimensional non-denaturing PAGE/SDS-PAGE. A simplified gel shift experiment displayed binding to fragmented calf thymus DNA. Nuclei were richest in poly(malate) binding proteins followed by cytoplasm and membranes. A protein of 370 kDa dissociated into 11 subunits of 11-29 kDa, indicative of a highly complex protein. This and other proteins displayed binding to nucleic acid in gel shift experiments. Poly(malate) is considered a structural and functional equivalent of long contiguous aspartate repeats in proteins of eukaryotes. PMID- 16476582 TI - No induction of anti-viral responses in human cell lines HeLa and MCF-7 when transfecting with siRNA or siLNA. AB - Gene silencing by RNAi and siRNAs has become a well-used tool for researchers. Because of its relatively small size, siRNA was originally thought to avoid activation of anti-viral responses. Recent reports demonstrating so-called "off target effects" are therefore alarming. One issue raised is that siRNA induces interferon-regulated genes at the transcriptional level. We characterize the anti viral responses of synthetic siRNA and in vitro-transcribed siRNA by measuring the mRNA levels of IFN-beta and OAS2 in HeLa cells. Transfections with both traditional and LNA-modified synthetic siRNA cause no anti-viral responses, whereas transfection with either long dsRNA or in vitro-transcribed siRNA leads to greater than 1000-fold induction of these genes. The lack of response was also demonstrated at the level of phosphorylated eIF2alpha, and measuring of IFN-beta by ELISA in cell culture media from the human cell line MCF-7. Altogether, transfection with synthetic siRNA does not induce anti-viral responses in these two cell lines. Our results reinforce the role of siRNA as an effective tool for reverse genetics and strengthen siLNA as a tool for future therapeutic applications. PMID- 16476583 TI - Role of Cbfa1/Runx2 in the fluid shear stress induction of COX-2 in osteoblasts. AB - Induction of cyclooxygenase-2 (COX-2) is thought to be important for the anabolic effects of mechanical loading. The transcription factor Cbfa1/Runx2 is essential for osteoblastic differentiation. We examined the role of Cbfa1 in the fluid shear stress (FSS) induction of COX-2 in MC3T3-E1 cells stably transfected with a COX-2 promoter-luciferase reporter. Cells were subjected to FSS for 30 min and returned to static culture (post-FSS). COX-2 mRNA and promoter activity peaked 0.5-1h and 2-3h, respectively, post-FSS. Mutation of the Cbfa1 consensus sequence at -267/-261 bp decreased the FSS fold-induction of luciferase activity by 50%. On electrophoretic mobility shift assay (EMSA), proteins binding to an oligonucleotide spanning the Cbfa1 site were supershifted by specific antibody to Cbfa1. FSS did not increase Cbfa1 binding on EMSA or Cbfa1 mRNA or protein levels. These data suggest that transcriptional activity of Cbfa1, independent of its level of expression, is necessary for maximal FSS induction of COX-2 in osteoblasts. PMID- 16476584 TI - Engineering functional artificial hybrid proteins between poplar peroxiredoxin II and glutaredoxin or thioredoxin. AB - The existence of natural peroxiredoxin-glutaredoxin hybrid enzymes in several bacteria is in line with previous findings indicating that poplar peroxiredoxin II can use glutaredoxin as an electron donor. This peroxiredoxin remains however unique since it also uses thioredoxin with a quite good efficiency. Based on the existing fusions, we have created artificial enzymes containing a poplar peroxiredoxin module linked to glutaredoxin or thioredoxin modules. The recombinant fusion enzymes folded properly into non-covalently bound homodimers or homotetramers. Two of the three protein constructs exhibit peroxidase activity, a reaction where the two modules need to function together, but they also display enzymatic activities specific of each module. In addition, mass spectrometry analyses indicate that the Prx module can be both glutathiolated or overoxidized in vitro. This is discussed in the light of the Prx reactivity. PMID- 16476585 TI - Trp-Arg-Trp-Trp-Trp-Trp antagonizes formyl peptide receptor like 2-mediated signaling. AB - Although formyl peptide receptor like 2 (FPRL2) has been regarded as an important classical chemoattractant receptor, its functional role and signaling pathway have not been fully investigated, because of the lack of its specific ligand. Recently F2L, a heme-binding protein fragment peptide, has been reported as an FPRL2-selective endogenous agonist. In the present study, we examined the effect of Trp-Arg-Trp-Trp-Trp-Trp-CONH2 (WRWWWW, WRW4), on F2L-induced cell signaling. WRW4 inhibited the activation of FPRL2 by F2L, resulting in the complete inhibition of intracellular calcium increase and chemotactic migration induced by F2L. WRW4 also completely inhibited F2L-induced NF-kappaB activation in FPRL2 transfected HEK293 cells. WRW4 specifically inhibited F2L-induced intracellular calcium increase and chemotactic migration in mature monocyte-derived dendritic cells, which express FPRL2 but not the other FPR family. Taken together, WRW4 is the first FPRL2 antagonist and is expected to be useful in the study of FPRL2 signaling and in development of drugs against FPRL2-related cellular responses. PMID- 16476586 TI - Simulating the pattern of right-hemisphere-damaged patients for the processing of the alternative metaphorical meanings of words: evidence in favor of a cognitive resources hypothesis. AB - Lately, many studies have suggested that communication impairments in brain damaged individuals might be explained--at least in part--in terms of cognitive resource allocation. Reproducing a clinical pattern in normal subjects by using a dual-task treatment might be a way of evaluating the role of cognitive resources in the right hemisphere's contribution to verbal communication. This study suggests that the RH's presumed specific contribution to the processing of the alternative metaphorical meanings of words is, at least to some extent, the expression of the RH's complementary, and necessary, contribution to the pool of attentional resources needed for the most effortful processing. PMID- 16476588 TI - Trends, complications, and mortality in peripheral vascular surgery. AB - BACKGROUND: The recent evolution in treatments for peripheral vascular disease has dated available mortality statistics for vascular intervention. Moreover, many of our current mortality statistics are derived from single-institution studies that are often not reflective of outcomes in general practice. To provide current and generalizable data regarding mortality and trends for peripheral vascular interventions, we examined two national data sets (Nationwide Inpatient Sample, 1998-2003, and National Hospital Discharge Survey, 1979-2003) and four states (New York, California, Florida, and New Jersey, 1998-2003). METHODS: Four procedures--abdominal aortic aneurysm repair (nonruptured), lower extremity revascularization, amputation, and carotid revascularization--were selected by cross-referencing International Classification of Diseases, 9th Revision, diagnostic and procedural codes. For significance, the t test was used for continuous variables, the chi2 test was used for dichotomous variables, and the chi2 test was used for mortality trends. RESULTS: From 1998 to 2003, there was a progressive decrease in the national per capita rate of amputations: 13.2% overall and 21.2% for major amputations (P < .0001). Nationally and regionally, mortality has only slightly declined. For lower extremity revascularization, after a sharp increase during the 1980s to 100,000 open procedures, the volume remained constant for 10 years and began to decline in 1998, reaching 70,000 cases in 2003. In contrast, since 1996, endovascular interventions have increased 40%. Mortality during the 1998 to 2003 period remained virtually stable at 1.5% to 2% for endovascular procedures and 3% to 4% for open procedures. The overall volume of abdominal aortic aneurysm repair has not changed substantially for the past 6 years; however, endovascular repair is now used for nearly half the cases (46.5% regional and 43.0% national). Mortality for open repair has not changed, remaining at approximately 5%, whereas for endovascular repair, mortality has declined from 2.6% in 2000 to less than 1.5% in 2003. After the rapid increase in open carotid revascularization in the early 1990s, the total volume has declined 5% nationally from 1998 to 2003. Regional data demonstrated an overall 12% reduction in carotid revascularization volume since 1998; this reduction was due to a 16% decline in open carotid revascularization. During this same period, the use of angioplasty-stent carotid revascularization doubled. Mortality for the open procedures is 0.5% and is significantly higher (2%-3%) for endovascular carotid revascularization. Stroke rates for endovascular carotid revascularization are also higher: 2.13% vs 1.28% for open procedures (P < .0001). CONCLUSIONS: Dramatic shifts in the management of peripheral vascular disease have occurred together with an overall decline in mortality. There seems to be a significant mortality advantage for endovascular as compared with traditional surgery except for carotid endarterectomy. The increasing safety of vascular interventions should be considered when deciding which patients to treat but with the caveat that endovascular interventions are not always safer than open repair. PMID- 16476589 TI - Thirty-day mortality statistics underestimate the risk of repair of thoracoabdominal aortic aneurysms: a statewide experience. AB - OBJECTIVE: The purpose of this study was to determine the 30-day and 365-day mortality for the repair of thoracoabdominal aortic aneurysms (TAA), when stratified by age, in the general population. These data provide clinicians with information more applicable to an individual patient than mortality figures from a single institutional series. METHODS: Data were obtained from the California Office of Statewide Health Planning and Development (OSHPD) for the years 1991 to 2002. These data were linked to the state death certificate file, allowing for continued information on the status of the patients after hospital discharge. All patients undergoing elective and ruptured TAA repair as coded by International Classification of Diseases, 9th Clinical Modification (ICD-9, CM) in California were identified. Patients aged <50 or >90 years old were excluded. We determined 30- and 365-day mortality and stratified our findings by decade of patient age (eg, 50 to 59). Demographics of elective and ruptured cases were also compared. RESULTS: We identified 1010 patients (797 elective, 213 ruptured) who underwent TAA repair. Mean patient ages were 70.0 (elective) and 72.1 years (ruptured). Men comprised 62% of elective and 68% of ruptured aneurysm patients, and 80% (elective) and 74% (ruptured) were white. Overall elective patient mortality was 19% at 30 days and 31% at 365 days. There was a steep increase in mortality with increasing age, such that elective 365-day mortality increased from about 18% for patients 50 to 59 years old to 40% for patients 80 to 89 years old. The elective case 31-day to 365-day mortality ranged from 7.8% for the youngest patients to 13.5%. Mortality for ruptured cases was 48.4% at 30 days and 61.5% at 365 days, and these rates also increased with age. CONCLUSIONS: Our observed 30-day mortality for TAA repairs is consistent with previous reports; however, mortality at 1 year demonstrates a significant risk beyond the initial perioperative period, and this risk increases with age. These data reflect surgical mortality for TAA repair in the general population and may provide more useful data for surgeons and patients contemplating TAA surgery. PMID- 16476591 TI - Age stratified, perioperative, and one-year mortality after abdominal aortic aneurysm repair: a statewide experience. AB - OBJECTIVE: The purpose of this study was to determine the in-hospital, 30-day, and 365-day mortality for the open repair of abdominal aortic aneurysms (AAAs), when stratified by age, in the general population. Age stratification could provide clinicians with information more applicable to an individual patient than overall mortality figures. METHODS: In a retrospective analysis, data were obtained from the California Office of Statewide Health Planning and Development (OSHPD) for the years 1995 to 1999. Out-of-hospital mortality was determined via linkage to the state death registry. All patients undergoing AAA repair as coded by International Classification of Diseases, 9th Revision (ICD-9) procedure code 38.44 and diagnosis codes 441.4 (intact) and 441.3/441.5 (ruptured) in California were identified. Patients <50 years of age were excluded. We determined in hospital, 30-day, and 365-day mortality, and stratified our findings by patient age. Multivariate logistic regression was used to determine predictors of mortality in the intact and ruptured AAA cohorts. RESULTS: We identified 12,406 patients (9,778 intact, 2,628 ruptured). Mean patient age was 72.4 +/- 7.2 years (intact) and 73.9 +/- 8.2 (ruptured). Men comprised 80.9% of patients, and 90.8% of patients were white. Overall, intact AAA patient mortality was 3.8% in hospital, 4% at 30 days, and 8.5% at 365 days. There was a steep increase in mortality with increasing age, such that 365-day mortality increased from 2.9% for patients 51 to 60 years old to 15% for patients 81 to 90 years old. Mortality from day 31 to 365 was greater than both in-hospital and 30-day mortality for all but the youngest intact AAA patients. Perioperative (in-hospital and 30-day) mortality for ruptured cases was 45%, and mortality at 1 year was 54%. CONCLUSIONS: There is continued mortality after the open repair of AAAs during postoperative days 31 to 365 that, for many patients, is greater than the perioperative death rate. This mortality increases dramatically with age for both intact and ruptured AAA repair. PMID- 16476592 TI - A decade of change in abdominal aortic aneurysm repair in the United States: Have we improved outcomes equally between men and women? AB - OBJECTIVES: Abdominal aortic aneurysm (AAA) repair has undergone vast changes in the last decade. We reviewed a national database to evaluate the effect on utilization of services and rupture rates. METHODS: From the Centers for Medicare Services (CMS), a 5% inpatient sample was obtained for 1994 to 2003 as beneficiary encrypted files (5% BEF) and as a limited data set file after 2001. Files were translated into Microsoft Access by using a custom program. Queries were performed using International Classification of Diseases (9th Revision) (ICD 9) diagnosis codes 441.3 (ruptured AAA) or 441.4 (non-ruptured AAA) and ICD-9 procedure codes 38.34, 38.36, 38.44, 38.64, 39.25, 39.52 for open, and 39.71 (available after October 2000) for endovascular repair. The 5% BEF totals were multiplied by 20 to calculate yearly volumes. Total cases were divided into the yearly CMS population of elderly Medicare recipients for repair rates per capita and are reported as cases per 100,000 elderly Medicare recipients. Statistics were performed using chi2, Student's t test, nonparametric tests, and multiple regression analysis; P < or = .05 was considered significant. RESULTS: Elective AAA repairs declined from 94.4/100,000 in 1994 to 87.7/100,000 in 2003. AAA rupture surgery declined from 18.7/100,000 (1994) to 13.6/100,000 (2003). Rupture repairs from 1994 to 2003 decreased by 29% for men and by 12% for women (P < .001). Rupture mortality has not changed, but the average is significantly higher for women at 52.8%, with men averaging 44.2% (P < .001). Mortality for elective AAA repair has decreased from 5.57% (1994) to 3.20% (2003) in men (P < .001) and from 7.48% (1994) to 5.45% (2003) in women (P < .001). Multivariate analysis demonstrated increasing age, female sex, and open surgery (vs endovascular) were significant predictors of elective and ruptured AAA repair mortality. For 2003 elective AAA repairs, the average length of stay was 6.9 days in men and 8.9 days in women (P < .01) For 2003, men were more likely to be discharged to home after rupture (32.9% of men vs 23.3% of women; P < .001) and elective repair (84.5% of men vs 70.1% of women; P < .001). CONCLUSIONS: Improvements in AAA management in the last decade have decreased aneurysm-related deaths and reduced the incidence of aneurysm ruptures, with a lower utilization of services. Women, however, continue to have a consistently higher mortality for open and ruptured AAA repair and are less likely to return to home after either. PMID- 16476594 TI - The value of motor evoked potentials in reducing paraplegia during thoracoabdominal aneurysm repair. AB - OBJECTIVE: Paraplegia after thoracoabdominal aortic aneurysm (TAAA) repair mainly occurs in patients with Crawford extent I and II. We assessed the impact of monitoring spinal cord integrity and the subsequent adjusted surgical maneuvers on neurologic outcome in repairs of type I and II TAAAs. METHODS: Surgical repair of TAAAs was performed in 112 consecutive patients with extent type I (n = 42) and type II (n = 70) aneurysms. The surgical protocol included cerebrospinal fluid drainage, moderate hypothermia, and left heart bypass with selective organ perfusion. Spinal cord function was assessed by means of monitoring motor evoked potentials (MEPs). Significant decreased MEPs always generated adjustments, including raising distal aortic and mean arterial pressure, reattachment of visible intercostal arteries, or endarterectomy of the excluded aortic segment with revascularization of back bleeding intercostal arteries. RESULTS: Motor evoked potential monitoring could be achieved in all patients. By maintaining a mean distal aortic pressure of 60 mm Hg, MEPs were adequate in 82% of patients. Increasing distal aortic pressure restored MEPs in all patients. In 19 patients (17%), MEPs decreased significantly during aortic cross-clamping because of critical spinal cord ischemia. MEPs returned in all patients after spinal cord blood flow was re-established except in three patients with type II TAAA in whom MEPs could not be restored, and absent MEPs at the end of the procedure corresponded with neurologic deficit. Delayed paraplegia developed in two patients owing to hemodynamic instability with insufficient mean arterial blood pressure to maintain adequate spinal cord perfusion. CONCLUSION: Monitoring MEPs is a highly reliable technique to assess spinal cord ischemia during TAAA repair. A surgical protocol including cerebrospinal fluid drainage, left heart bypass, and monitoring of MEPs can reduce the paraplegia rate significantly. Adjusted hemodynamic and surgical strategies induced by changes in MEPs could restore spinal cord ischemia in most patients, preventing early and late paraplegia in all type I patients. In type II patients, early paraplegia occurred in 4.2% and delayed neurologic deficit in 2.9%. Despite all available measures, complete prevention of paraplegia in type II aneurysms seems to be unrealistic. PMID- 16476595 TI - Endograft exclusion of acute and chronic descending thoracic aortic dissections. AB - OBJECTIVES: To analyze the results of endograft exclusion of acute and chronic descending thoracic aortic dissections (Stanford type B) with the AneuRx (n = 5) and Talent (n = 37) thoracic devices and to compare postoperative outcomes of endograft placement acutely (<2 weeks) and for chronic interventions. METHODS: Patients treated for acute or chronic thoracic aortic dissections (Stanford type B) with endografts were included in this study. All patients (n = 42) were enrolled in investigational device exemption protocols from August 1999 to March 2005. Three-dimensional computed tomography reconstructions were analyzed for quantitative volume regression of the false lumen and changes in the true lumen over time (complete >95%, partial >30%). RESULTS: Forty-two patients, all of whom had American Society of Anesthesiologists (ASA) risk stratification > or =III and 71% with ASA > or = IV, were treated for Stanford type B dissections (acute = 25, chronic = 17), with 42 primary and 18 secondary procedures. All proximal entry sites were identified intraoperatively by intravascular ultrasound (IVUS). The procedural stroke rate was 6.7% (4/60), with three posterior circulation strokes. Procedural mortality was 6.7% (4/60). The left subclavian artery was occluded in 11 patients (26%) with no complaints of arm ischemia, but there was an association with posterior circulation strokes (2/11) (18%). No postoperative paraplegia was observed after primary or secondary intervention. Complete thrombosis of the false lumen at the level of endograft coverage occurred in 25 (61%) of 41 patients < or =1 month and 15 (88%) of 17 patients at 12 months. Volume regression of the false lumen was 66.4% (acute) and 91.9% (chronic) at 6 months. Lack of true lumen volume (contrast) increase and increasing false lumen volume (contrast) suggests continued false lumen pressurization and the need for secondary reintervention. Thirteen patients (31%) required 18 secondary interventions for proximal endoleaks in 6, junctional leaks in 3, continued perfusion of the false lumen from distal re-entry sites in 3, and surgical conversion in 4 for retrograde dissection. CONCLUSIONS: Preliminary experience with endografts to treat acute and chronic dissections is associated with a reduced risk of paraplegia and lower mortality compared with open surgical treatment, the results of medical treatment alone, or a combination. PMID- 16476596 TI - Contrast-enhanced Duplex surveillance after endovascular abdominal aortic aneurysm repair: improved efficacy using a continuous infusion technique. AB - INTRODUCTION: Currently, postoperative endoleak surveillance after endovascular aortic aneurysm repair (EVAR) is primarily done by computed tomography (CT). The purpose of this study was to determine the efficacy of contrast-enhanced ultrasonography scans to detect endoleaks by using a novel infusion method and compare these findings with those of CT angiography (CTA). METHODS: Twenty male patients (mean age, 70.4 years) underwent surveillance utilizing both CTA and contrast-enhanced color Duplex imaging. One 3-mL vial of Optison (Perfluten Protein A microspheres for injection) and 57 mL normal saline, for a total of 60 mL, were administered to each patient as a continuous infusion at 4 mL/min via a peripheral vein. Each study was optimized with harmonic imaging, and a reduced mechanical index of 0.4 to 0.5, compression of 1 to 3, and a focal zone below the aorta to minimize microsphere rupture. One minute was allowed from the time of infusion to the appearance of contrast in the endograft. Flow was evaluated within the lumen of the graft and its components, as was the presence or absence of endoleaks. Findings were compared with standard color-flow Duplex imaging and CT utilizing CTA reconstruction protocols. RESULTS: All patients evaluated had modular endografts implanted for elective aneurysm repair. Contrast-enhanced duplex scans identified nine endoleaks: one type I and eight type II. No additional endoleaks were seen on CTA. However, CTA failed to recognize three type II endoleaks seen by contrast-enhanced ultrasound. The continuous infusion method allowed for longer and more detailed imaging. An average of 46.8 mL of the contrast infusion solution was used per patient. CONCLUSIONS: Contrast enhanced Duplex ultrasonography accurately demonstrates endoleaks after EVAR and may be considered as a primary surveillance modality. Continuous infusion permits longer imaging time. PMID- 16476598 TI - Screening and ultrasound surveillance of large abdominal aortic aneurysms do not improve suitability for endovascular repair. AB - OBJECTIVE: The effect of population screening for abdominal aortic aneurysm (AAA) disease on morbidity and mortality has been comprehensively studied and reported. However, the effect of early AAA detection on suitability for endovascular aneurysm repair (EVAR) remains unknown. Considering the importance of such an effect on future health economics, we sought to assess the possible effect of AAA ultrasound surveillance on suitability for EVAR. METHODS: This was a prospective cohort study. From January 2002 to August 2003, consecutive AAA patients selected for open elective repair were placed into one of two groups according to mode of presentation. The first group included patients referred from a local well established AAA ultrasound screening and surveillance program (ultrasound surveillance [AAA-S] group). The second group included patients referred from neighboring unscreened regions with incidentally diagnosed AAA (incidental [AAA I] group). All patients underwent preoperative computed tomographic angiography. By using three-dimensional reconstruction software, computed tomographic images were assessed by two blinded observers for suitability for EVAR by using the criteria for a modular endovascular device. RESULTS: Of 74 patients included in the study, 41 were in the AAA-S group, and 31 were in the AAA-I group. The median aneurysm diameter was 72.3 mm (range, 50.7-83.7 mm) for AAA-I and 65 mm (range, 50.7-79.2 mm) for AAA-S (P < .47). Suitability for EVAR was 41% in the AAA-S group and 45% in the AAA-I group (P < .47). CONCLUSIONS: Early detection and surveillance of AAA does not seem to increase suitability for EVAR. Suitability for EVAR seems to be determined early on in an aneurysm's life. On the basis of current device technology, referral for intervention from an AAA surveillance program may need to be initiated at a size well below 5.5 cm if an increase in EVAR suitability is to be expected. PMID- 16476600 TI - Totally percutaneous aortic aneurysm repair: experience and prudence. AB - OBJECTIVE: Totally percutaneous aortic aneurysm repair has been shown to be technically feasible, with low morbidity. Complications from percutaneous arterial closure are not insignificant, however, and can be fatal. We sought to evaluate our experience with this technique, compare it with the published literature, and identify factors associated with conversion to open repair and complications. METHODS: A retrospective review of a prospectively collected database was performed. All patients who underwent percutaneous closure of large bore-sheath (>12F) access sites with off-label use of a suture-mediated closure device (Prostar XL) between December 2002 and August 2005 were reviewed. Outcome measures evaluated were rates of technical success, conversion to open femoral arterial repair, and complications. Axial diameter measurements of the accessed vessels were assessed with computed tomographic (CT) angiography both before and after the procedure. Patient variables were compared by using chi2, Fisher exact, and paired and independent samples t tests where appropriate. The mean follow-up interval was 1.5 years. RESULTS: During the study period, 49 patients underwent percutaneous closure of 79 large-bore-sheath access sites after successful endovascular aneurysm repair. Seven patients (14%) were morbidly obese (body mass index >35 kg/m2). Successful closure was achieved in 74 access sites (93.7%). Percutaneous closure was unsuccessful in five access sites (6.3%), all of which required open femoral repair at the same setting. Two converted patients experienced complications (4.1%): one retroperitoneal hematoma requiring transfusion of blood products and one iliac artery injury leading to death from myocardial infarction. Both of these patients were morbidly obese. Both complications occurred after closure of larger than 20F sheath sites. Morbid obesity and sheath size greater than 20F were associated with a significantly increased complication rate (P = .02 and P = .01, respectively). No thrombotic or infectious complications occurred in this series. Upon comparison of preoperative and postoperative CT angiograms, one (1.3%) small pseudoaneurysm was detected. No arteriovenous fistulas or hematomas larger than 3 cm were detected. The pseudoaneurysm occurred after closure of a 20F sheath access site. There were no significant differences in minimum intraluminal (7.38 +/- 1.8 vs 7.48 +/- 1.8) or maximum extraluminal (11.25 +/- 2.8 vs 12.02 +/- 2.7) diameters between preoperative and postoperative CT angiograms, respectively. CONCLUSIONS: Totally percutaneous aortic aneurysm repair is technically feasible in most cases, with no effect on the luminal diameter of the accessed femoral artery. Complications occur more often in morbidly obese patients and with sheaths larger than 20F. These complications can be minimized with meticulous technique and good patient selection. The capability for expeditious open femoral arterial repair is mandatory with this approach. PMID- 16476601 TI - Long-term outcome after Talent endograft implantation for aneurysms of the abdominal aorta: a multicenter retrospective study. AB - BACKGROUND: The development of newer-generation endografts for the endovascular treatment of abdominal aortic aneurysms has resulted in considerable improvements in clinical performance. However, long-term outcome data are still scarce. To assess long-term clinical and radiographic outcomes after use of the Talent stent graft, a retrospective analysis was performed that was based on 165 patients treated with this endograft in Germany between October 1996 and December 1998. METHODS: Data were collected according to the recommendation of the ad hoc committee for standardized reporting practices in vascular surgery and were evaluated statistically by using univariate and multivariate analyses. RESULTS: A total of 165 patients were treated with a Talent endograft in 9 German centers before December 31, 1998. Most were asymptomatic (94.5%), male (97.6%), and treated with a bifurcated graft (86.7%). Two patients (1.2%) died within 30 days, and 28 (17%) died during the follow-up period. The cause of death was aneurysm rupture in one case. Survival was 95.4% +/- 1.7% at 1 year, 89% +/- 2.6% at 2 years, 78.1% +/- 3.6% at 5 years, and 76.2% +/- 4.1% at 7 years. Patients classified as American Society of Anesthesiologists grade IV had a significantly lower survival rate (24.9%) than those classified as American Society of Anesthesiologists grade II and III (91.9% and 77.3%). During a mean follow-up period of 53.2 +/- 20.1 months (range, 1-84 months), 47 secondary procedures were performed in 31 patients (18.8%). Kaplan-Meier estimates showed a freedom from secondary intervention of 94.7% +/- 1.8%, 81.7% +/- 3.3%, and 77.4% +/- 3.6% at 1, 3, and 7 years, respectively. The reason for secondary treatment was endograft thrombosis in 10 patients (6.1%), persisting primary endoleak in 9 (5.5%), late secondary endoleak in 6 (3.6%), graft migration in 3 (1.8%), aneurysm rupture in 2 (1.2%), and graft infection in 1 (0.6%). Device migration (> or =10 mm) occurred in seven patients (4.2%). Other graft changes, such as graft kinking (n = 4; 2.4%), fracture of metallic stents (n = 2; 1.2%), erosion of the longitudinal bar (n = 2; 1.2%), or modular component separation (n = 1; 0.6%), were rare. Follow-up computed tomographic imaging revealed a decrease of the maximum aneurysm sac diameter (>5 mm) in 106 (64.2%) patients and an increase in 14 (8.5%) patients. The mean aneurysm diameter significantly decreased (P < .001). Of the factors recorded at baseline, only endoleaks showed a significant correlation with the risk of aneurysm increase during follow-up (P < .001). Adverse anatomy (neck diameter >28 mm, neck length <15 mm, and '5 patent aortic branches) did not adversely influence the aneurysm shrinkage rate, the risk for a secondary procedure, or the clinical success rate. A significantly higher rate of clinical success (P < .05) was observed in patients older than 65 years of age. CONCLUSIONS: Implantation of the Talent endograft device is a safe and effective alternative to open surgery for exclusion of abdominal aortic aneurysm. In comparison with first-generation grafts, the device showed superior durability for as long as 5 to 7 years after implantation. Even if prototypes of the Talent device were implanted in this study, the graft was also successfully used in most patients, even in those with adverse anatomy. Because improvements of the endograft have been made to address connecting bar breaks, a lower incidence of graft limb occlusion can be expected in the future. PMID- 16476603 TI - Defining the high-risk patient for carotid endarterectomy: an analysis of the prospective National Surgical Quality Improvement Program database. AB - BACKGROUND: Carotid endarterectomy (CEA) is the gold standard for the treatment of carotid stenosis, but carotid angioplasty and stenting has been advocated in high-risk patients. The definition of such a population has been elusive, particularly because the data are largely retrospective. Our study examined results for CEA in the National Surgical Quality Improvement Program database (both Veterans Affairs and private sector). METHODS: National Surgical Quality Improvement Program data were gathered prospectively for all patients undergoing primary isolated CEA during the interval 2000 to 2003 at 123 Veterans Affairs and 14 private sector academic medical centers. Study end points included the 30-day occurrence of any stroke, death, or cardiac event. A variety of clinical, demographic, and operative variables were assessed with multivariate models to identify risk factors associated with the composite (stroke, death, or cardiac event) end point. Adjudication of end points was by trained nurse reviewers (previously validated). RESULTS: A total of 13,622 CEAs were performed during the study period; 95% were on male patients, and 91% of cases were conducted within the Veterans Affairs sector. The average age was 68.6 +/- 0.1 years, and 42.1% of the population had no prior neurologic event. The composite stroke, death, or cardiac event rate was 4.0%; the stroke/death rate was 3.4%. Multivariate correlates of the composite outcome were (odds ratio, P value) as follows: deciles of age (1.13, .018), insulin-requiring diabetes (1.73, <.001), oral agent controlled diabetes (1.39, .003), decade of pack-years smoking (1.04, >.001), history of transient ischemic attack (1.41, >.001), history of stroke (1.51, >.001), creatinine >1.5 mg/dL (1.48, >.001), hypoalbuminemia (1.49, >.001), and fourth quartile of operative time (1.44, >.001). Cardiopulmonary comorbid features did not affect the composite outcome in this model. Regional anesthesia was used in 2437 (18%) cases, with a resultant relative risk reduction for stroke (17%), death (24%), cardiac event (33%), and the composite outcome (31%; odds ratio, 0.69; P = .008). CONCLUSIONS: Carotid endarterectomy results across a spectrum of Veterans Affairs and private sector hospitals compare favorably to contemporary studies. These data will assist in selecting patients who are at an increased risk for adverse outcomes. Use of regional anesthetic significantly reduced perioperative complications in a risk-adjusted model, thus suggesting that it is the anesthetic of choice when CEA is performed in high-risk patients. PMID- 16476605 TI - Carotid artery stenting in octogenarians is associated with increased adverse outcomes. AB - BACKGROUND: Carotid artery stenting is an increasingly common endovascular treatment of carotid artery stenosis advocated in high-risk patients despite reports of increased adverse periprocedural outcomes in patients aged >80 years. We sought to evaluate our single institution experience with octogenarians and whether they have an increased incidence of major complications with carotid artery stenting. METHODS: Three hundred eighty-six patients, including 260 patients from 10 regulatory trials, who underwent carotid artery stenting between June 1996 and March 2004 for symptomatic or asymptomatic carotid stenosis were reviewed from a prospectively maintained database. Periprocedural (< or =30 days after carotid artery stenting) cerebrovascular accident, transient ischemic attack, myocardial infarction, and death outcomes were compared between 87 octogenarians and 295 nonoctogenarians. Univariate and multivariate analysis was performed for confounding factors. Kaplan-Meier analysis of stroke and death outcomes was performed for a 1-year follow-up. RESULTS: All adverse outcomes were significantly higher in octogenarians compared with younger patients: 30-day stroke rate, 8.0% vs 2.7% (P = .02); 30-day stroke, myocardial infarction, or death, 9.2% vs 3.4% (P = .02). Cohorts were similar in terms of gender, comorbidities, antiplatelet medications, symptomatic status, and use of cerebral protection. Octogenarians had a greater incidence of contralateral internal carotid artery occlusion (26% vs 12%, P = .001), atrial fibrillation (21% vs 8%, P = .001), and congestive heart failure (28% vs 15%, P = .007), but a lower incidence of hypercholesterolemia (53% vs 72%, P = .001) and active smoking (8% vs 24%, P = .001). Multivariate analysis of 30-day major adverse outcomes demonstrated an association between age > or =80 and adverse outcome (odds ratio, 2.85; P = .043) as well as a protective effect of the preprocedural use of aspirin (odds ratio, 0.30, P = .027). At 1-year follow-up, only 75% of octogenarians and 87% of nonoctogenarians were free from stroke, myocardial infarction, or death (P = 005, Kaplan-Meier analysis). CONCLUSIONS: Octogenarians undergoing carotid artery stenting are at higher risk than nonoctogenarians for periprocedural complications, including neurologic events and death. Major event free survival at 1 year is also significantly better in nonoctogenarians. These risks should be weighed when considering carotid stenting in elderly patients. PMID- 16476606 TI - Management of in-sent restenosis after carotid artery stenting in high-risk patients. AB - BACKGROUND: Carotid artery stenting (CAS) has emerged as an acceptable treatment alternative in patients with carotid bifurcation disease. Although early results of CAS have been promising, long-term clinical outcomes remain less certain. We report herein the frequency, management, and clinical outcome of in-stent restenosis (ISR) after CAS at a single academic institution. METHODS: Clinical records of 208 CAS procedures in 188 patients with carotid stenosis of 80% or greater, including 48 (26.5%) asymptomatic patients, during a 42-month period were analyzed. Follow-up serial carotid duplex ultrasound scans were performed. Selective angiography and repeat intervention were performed when duplex ultrasound scans showed 80% or greater ISR. Treatment outcomes of ISR interventions were analyzed. RESULTS: Over a median 17-month follow-up, 33 (15.9%) ISRs of 60% or greater were found, according to the Doppler criteria. Among them, seven patients (3.4%) with a mean age of 68 years (range, 65-87 years) developed high-grade ISR (> or =80%), and they all underwent further endovascular interventions. Six patients with high-grade ISR were asymptomatic, whereas one remaining patient presented with a transient ischemic attack. Five of seven ISRs occurred within 12 months of CAS, and two occurred at 18 months' follow-up. Treatment indications for initial CAS in these seven patients included recurrent stenosis after CEA (n = 4), radiation-induced stenosis (n = 1), and high-cardiac-risk criteria (n = 2). Treatment modalities for ISR included balloon angioplasty alone (n = 1), cutting balloon angioplasty alone (n = 4), cutting balloon angioplasty with stent placement (n = 1), and balloon angioplasty with stent placement (n = 1). Technical success was achieved in all patients, and no periprocedural complications occurred. Two patients with post-CEA restenosis developed restenosis after ISR interventions, both of whom were successfully treated with cutting balloon angioplasty at 6 and 8 months. The remaining five patients showed an absence of recurrent stenosis or symptoms during a mean follow up of 12 months (range, 3-37 months). By using the Kaplan-Meier analysis, the freedom from 80% or greater ISR after CAS procedures at 12, 24, 36, and 42 months was 97%, 97%, 96%, and 94%, respectively. CONCLUSIONS: Our study showed that ISR after CAS remains uncommon. Successful treatment of ISR can be achieved by endovascular interventions, which incurred no instance of periprocedural complications in our series. Patients who developed ISR after CEA were likely to develop restenosis after IRS intervention. Diligent ultrasound follow-up scans are important after CAS, particularly in patients with post-CEA restenosis. PMID- 16476609 TI - Remote endarterectomy: lessons learned after more than 100 cases. AB - BACKGROUND: Bypass procedures have been the mainstay of treatment of extensive external iliac and superficial femoral artery (SFA) occlusive disease, particularly total occlusions. Since the early 1990s, reports from Europe have espoused the virtues of endarterectomy of the superficial femoral and iliac arteries from a small groin incision, but adoption in the United States has been limited. Over the past 4 years, we have explored the technical challenges and durability of this procedure and report our findings. METHODS: Remote endarterectomy from an inguinal incision was the primary treatment option for all patients considered surgical candidates for vascular reconstruction of the external iliac and superficial femoral arteries. All data were entered into an outcomes database prospectively and reviewed retrospectively. After the procedure, duplex ultrasound surveillance was performed quarterly the first year and semi-annually thereafter. RESULTS: Remote endarterectomy was the planned procedure in 133 patients. The mean age was 68 years, 68% were men, and 31% were diabetic. The indications for the procedure were claudication in 57% and limb salvage in 43%. In 16 patients (12%), technical issues precluded the completion of the remote endarterectomy and a bypass was performed. Successful retrograde iliac endarterectomy was performed in 7 patients, SFA endarterectomy in 105 patients, and combined retrograde iliac and antegrade SFA in 5 patients. The average duration of the procedure was 162 minutes +/- 69 minutes (SD). Half of the patients were discharged on the first postoperative day, and the average length of stay was 2.52 days. The mean follow-up was 19 months, with a primary patency of 70% at 30 months by life-table analysis. Limb salvage was 94%. CONCLUSIONS: Remote endarterectomy is a viable and durable alternative to standard bypass procedures. It has equivalent patency to published results of bypass or endovascular procedures of the external iliac and superficial femoral arteries and may soon replace bypass as the preferred procedure for long-segment occlusions of these vessels. PMID- 16476607 TI - Heparin-bonded expanded polytetrafluoroethylene vascular graft for femoropopliteal and femorocrural bypass grafting: 1-year results. AB - OBJECTIVE: Several prosthetic materials have been used for femoropopliteal bypass grafting in patients with peripheral vascular disease in whom a venous bypass is not possible. Expanded polytetrafluoroethylene (ePTFE) is the most commonly used, but patency results have not always equaled those achieved with vein, especially in below-knee reconstructions. This study assessed the performance of a new heparin-bonded ePTFE vascular graft that was designed to provide resistance to thrombosis and thereby decrease early graft failures and possibly prolong patency. METHOD: From June 2002 to June 2003, 86 patients (62 men and 24 women; mean age, 70 years; 99 diseased limbs) were enrolled prospectively in a nonrandomized, multicenter study of the heparin-bonded ePTFE graft. Fifty-five above-knee and 44 below-knee (including 21 femorocrural) procedures were performed. Follow-up evaluations consisted of clinical examinations, ultrasonographic studies, and distal pulse assessments. Patency and limb salvage rates were assessed by using life-table analyses. RESULTS: All grafts were patent immediately after implantation. There were no graft infections or episodes of prolonged anastomotic bleeding. During the 1-year follow-up, 10 patients died, 15 grafts occluded, and 5 major amputations were performed. The overall primary and secondary 1-year patency rates were 82% and 97%, respectively. The limb salvage rate in patients with critical limb ischemia (n = 41) was 87%. Primary patency rates according to bypass type were 84%, 81%, and 74% for above-knee femoropopliteal, below-knee femoropopliteal, and femorocrural bypasses, respectively; the corresponding secondary patency rates were 96%, 100%, and 100%. CONCLUSIONS: In this study, the heparin-bonded ePTFE graft provided promising early patency and limb salvage results, with no device-related complications, in patients with occlusive vascular disease. Longer-term and randomized studies are warranted to determine whether this graft provides results superior to those achieved with other prostheses, especially in patients at increased risk of early graft failure, such as those undergoing below-knee bypass and those with poor run off or advanced vascular disease. PMID- 16476612 TI - Digital venous photoplethysmography in the seated position is a reproducible noninvasive measure of lower limb venous function in patients with isolated superficial venous reflux. AB - BACKGROUND: The value of photoplethysmography (PPG) has been questioned because of a lack of reproducibility. We performed this study to determine whether new digital technology has improved the reproducibility of PPG in the noninvasive assessment of lower limb venous function in patients with isolated superficial venous reflux. METHODS: This was a prospective study of 140 legs in 110 patients (65% female; median age [interquartile range], 45 years [36-59.25 years]; CEAP clinical grade C2/3, n = 114; C4-6, n = 26) who underwent repeated digital PPG measurements of refilling time (RT) in both the sitting and standing position after standard exercise regimens by a single observer. RT was measured in all patients 2 to 5 minutes apart and in a randomly selected subgroup of 30 patients (38 limbs) 1 to 2 weeks apart. RT variability was assessed by using Bland and Altman's coefficient of repeatability (CR-RT), where the CR-RT was 1.96 times the standard deviation of the mean difference in RT between two tests. Venous duplex scanning of both the deep and superficial veins was also performed, and a reverse flow of greater than 0.5 seconds was considered abnormal. Only patients with isolated superficial venous reflux were included in the study. RESULTS: The CR-RT of the tests on 140 limbs performed 2 to 5 minutes apart was 10 seconds overall, 3 seconds for RT up to 10 seconds, and 16 seconds for RT between 20 and 40 seconds. The CR-RT of the 38 tests performed 1 to 2 weeks apart was also 10 seconds. No systematic variation due to a nonrandom error was found between the measurements performed either 2 to 5 minutes or 1 to 2 weeks apart. CONCLUSIONS: Digital PPG performed in the seated position in patients with isolated superficial venous reflux provides a reproducible method for the noninvasive assessment of lower limb venous function for both clinical and research purposes. However, the variation in precision of RT with the magnitude of the measurement must be taken into account when results are interpreted in individual patients. PMID- 16476610 TI - Presentation of the patient with recurrent varices after surgery (REVAS). AB - AIM: To identify in patients with recurrent varices after surgery (REVAS) the clinical, etiologic, anatomic, and pathophysiologic patterns according to the CEAP classification, as well as the site, source, causes of recurrence, and contributory factors by using the REVAS classification. METHODS: Centers from eight countries enrolled patients with superficial vein reflux that had had a previous operation. A physical examination and a duplex ultrasound scan were performed at the first visit. This was repeated between 2 to 8 weeks after by the same physician and by another physician within the same time frame. The perforator, deep, and superficial veins systems as well as their accessories and tributaries were examined. A form based on the CEAP and the REVAS classification was used and the data were entered in a customized database. RESULTS: Fourteen institutions enrolled 170 patients (199 lower limbs) in 1 year. Their mean age was 56 years, and 69% were women. Most of them had undergone one surgical procedure before enrollment (76.6%). Most had varicose veins and swelling (70.9%), and the rest had skin damage (29.1%). More than 90% had primary etiology. The saphenofemoral junction (47.2%) and leg perforators (54.7%) were the areas most often involved by recurrent reflux. Reflux in deep veins was detected in 27.4%. Class 2 (varicose veins) alone was present in 24.6% of limbs, two classes were present in 43%, and three in 24%. Neovascularization was as frequent as technical failure (20% vs 19%); both were seen in 17%. In 35%, the cause was uncertain or unknown. When recurrence occurred at a different site, development of reflux in new sites was found in 32% of limbs. Of the contributing factors, family history and lifestyle had the highest prevalence. Women had significantly more procedures than men, despite a clear trend toward more severe disease in the latter. CONCLUSIONS: Most patients were symptomatic with several clinical forms of presentation. The REVAS classification, together with CEAP, gives significant and more appropriate information for evaluating and following up patients with chronic venous disease who have had an intervention. PMID- 16476613 TI - A biologic basis for asymmetric growth in descending thoracic aortic aneurysms: a role for matrix metalloproteinase 9 and 2. AB - OBJECTIVE: This study was undertaken to define matrix metalloproteinase (MMP) expression in the anterior and posterior wall of descending thoracic aortic aneurysms (TAAs) and correlate it with specific computed tomography (CT) image sites within the descending thoracic aorta. METHODS: Serial CT images of patients with TAAs were compared with age- and gender-matched normal descending thoracic aortas at levels T4-T12. The mean circumference of the TAAs was 153 mm (n = 12) and 148 mm (n = 11) at T8 and T10, respectively, compared with 75 mm (n = 12) and 75 mm (n = 10) in controls (P < .001). Aortic tissue was collected from a separate set of eight patients undergoing descending TAA resection (processed < or =12 hours of excision) and six cadavers (processed < or =24 hours of death). Tissue collected between the intercostals arteries was defined as posterior wall, and directly opposite was the anterior wall. MMP-9 and MMP-2 messenger RNA (mRNA) extracted from aortic tissue was analyzed by quantitative real time polymerase chain reaction (PCR) and normalized to beta-actin. Immunohistochemistry was performed for MMP-9 and MMP-2. CT aortic measurements and MMP expression were compared by t tests and analysis of variance, respectively. RESULTS: The ratio of arc distance between the intercostals on the posterior wall to total aortic circumference was 0.14 in healthy controls compared with 0.08 in TAAs at vertebral level T8 (P = .001). At T10, the ratio was 0.15 in healthy controls compared with 0.11 in TAAs (P = .001). MMP-9 expression in TAAs was 4.3-fold higher in the anterior wall compared with the posterior wall (P = .03). Conversely, MMP-2 expression in TAAs was 3.2-fold higher in the posterior wall compared with the anterior wall (P = .008). MMP expression was not detected in control cadaver aortas. CONCLUSION: Anterior walls of expanding TAAs grow at a greater rate than the posterior wall, as determined from the lower ratio of intercostal arc distance to total circumference in TAAs. Differential MMP expression appears to be a biologic marker for asymmetric growth in the TAA wall. CLINICAL RELEVANCE: The pathogenesis of thoracic aortic aneurysms (TAAs) is poorly understood. Multiple lines of evidence suggest that matrix metalloproteinases (MMPs), a family of enzymes, are important in aneurysm development. Earlier experiments documented a regional variation of MMP-9 in stimulated rodent aortas, with production greater in the abdominal aorta compared with the thoracic aorta. The present study extends that observation and documents asymmetric aneurysm development in the TAA wall, with increased anterior wall growth in correlation to increased MMP-9 production. An improved understanding of the mechanisms by which MMP production is regulated is critical. PMID- 16476614 TI - The wrapping method using biodegradable felt strips has a preventive effect on the thinning of the aortic wall: experimental study in the canine aorta. AB - OBJECTIVES: Wrapping methods have been widely used to reinforce the anastomotic site in vascular surgery; however, postoperative changes in the aortic wall wrapped by nonbiodegradable felt have not been well characterized. The purposes of this investigation are to elucidate the sequelae of wrapping with nonbiodegradable felt on the aortic wall and to modify those changes by using biodegradable felt with or without basic fibroblast growth factor (bFGF). METHODS: The descending thoracic aortas of 15 beagles were wrapped with three different materials: nonbiodegradable polytetrafluoroethylene (PTFE) felt, biodegradable polyglycol acid (PGA) material, and PGA with 100 microg bFGF (n = 5 in each group). The descending thoracic aorta was resected after 3 months. The thickness of the aortic wall, vessel density in the media and the adventitia, and the wall strength were assessed. Untreated native aortic wall served as a normal control. RESULTS: The thickness of the media of the PTFE group was lower than that of the PGA + bFGF group (66% +/- 5% vs 85% +/- 6% of control, P < .05). The adventitia-media ratio in the PTFE group decreased compared with controls (59.1% of normal, P < 0.05), whereas those in the PGA and PGA + bFGF groups increased (172.1% and 189.6% of normal, respectively, P < .01). The collagen-smooth muscle ratio in the media was higher in the PTFE group than in the controls (0.14 +/- 0.02 vs 0.07 +/- 0.01, P < .01). The number of vessels in the adventitia was higher in the PGA + bFGF group than those in PTFE or PGA groups (29.6 +/- 2.5/mm2 vs 6.4 +/- 0.8/mm2, 19.0 +/- 1.1/mm2, P < .01). The PGA + bFGF group demonstrated larger failure force than the PTFE group (4.0 +/- 0.3 kgf vs 1.6 +/- 0.3 kgf, P < .01). The failure stress in the PGA and PGA + bFGF groups was larger than that in PTFE group (PTFE:PGA + bFGF = 5.3 +/- 0.9 x10(2) kPa:11.7 +/- 1.7 x 10(2) kPa, P < .01; PTFE:PGA = 5.3 +/- 0.9 x 10(2) kPa:11.2 +/- 1.2 x 10(2) kPa, P < .05). CONCLUSION: The aortic wall wrapped with nonbiodegradable PTFE felt showed a reduced thickness and diminished vessels in the adventitia. Biodegradable felt (PGA), with or without bFGF, modified these histologic changes. The vessel-rich thickened adventitia, after wrapping by PGA with bFGF, was associated with increased aortic wall strength. CLINICAL RELEVANCE: This investigation was conducted in an attempt to elucidate mechanisms underlying the occurrence of late postoperative false aneurysm after aortic surgery. We hypothesized that sustaining compression of the aorta by the felt strip may cause structural derangement and local ischemia on the aortic wall. We used a simple wrapping of the aorta with a felt strip rather than a felt strip at anastomotic sites to simplify the experimental model and to exclude confounding factors brought by technical inconsistency on the surgical anastomosis. We further attempted to find a clue for preventing adverse effects of wrapping with a conventional felt strip. Practically, we pursued a possible application of a biodegradable felt strip to aortic wrapping in our experimental model before we proceed in a clinical application of the new material. PMID- 16476615 TI - Neutrophil antimicrobial peptide alpha-defensin causes endothelial dysfunction in porcine coronary arteries. AB - BACKGROUND: Defensins are cysteine-rich cationic polypeptides released from neutrophils that exhibit powerful antimicrobial activities. Because inflammation, including neutrophil infiltration and release of defensins, may play an important role in atherosclerosis and other vascular diseases, we determined whether alpha defensin could cause endothelial dysfunction, a major initial event of atherosclerosis, in porcine coronary arteries. METHODS: Porcine coronary arteries were sliced into 5-mm rings and treated with different concentrations of human recombinant alpha-defensin for 24 hours. Vasomotor reactivity was studied by using a myograph system. Levels of superoxide anion were detected by the lucigenin-enhanced chemiluminescence method. Endothelial nitric oxide synthase (eNOS) messenger RNA (mRNA) and protein levels were determined by real-time polymerase chain reaction and immunohistochemistry analysis, respectively. RESULTS: Endothelium-dependent relaxation in response to bradykinin was significantly reduced by 40% for the rings treated with 1500 nM of alpha-defensin compared with controls (P< .05). Vessel contractility in response to the thromboxane A2 analogue U46619 and endothelium-independent relaxation in response to sodium nitroprusside were not affected with defensin treatment. In addition, the superoxide anion level at the endothelial layer of porcine coronary artery rings was significantly increased by 80% in the defensin-treated (1500 nM) vessels compared with controls (P< .05). Furthermore, the eNOS mRNA levels in endothelial cells isolated from the cultured rings treated with defensin (1500 nM) were significantly decreased by 27% compared with controls (P< .05). Immunoreactivity of eNOS in the defensin-treated vessel rings was also substantially reduced. CONCLUSIONS: Defensin reduces the endothelium-dependent vasorelaxation. This effect is associated with increased superoxide radical production and decreased eNOS expression in porcine coronary arteries. CLINICAL RELEVANCE: Inflammation is an important mechanism of atherosclerosis and other vascular diseases. The roles and interactions of biomediators released from inflammatory cells are not fully understood, however. This study provides new information about effects and potential molecular mechanisms of a major neutrophil releasing factor, alpha-defensin, on endothelial dysfunction of porcine coronary arteries. Thus, targeting alpha-defensin and its associated molecular mechanisms may become a new strategy to prevent vascular diseases. PMID- 16476616 TI - Synthetic smooth muscle cell phenotype is associated with increased nicotinamide adenine dinucleotide phosphate oxidase activity: effect on collagen secretion. AB - OBJECTIVE: Smooth muscle cells (SMCs) from prosthetic vascular grafts secrete higher levels of collagen than aortic SMCs under basal conditions and during incubation with oxidized low-density lipoprotein. We postulated that reactive oxygen species (ROS) contributed to the observed difference. The objective of this study was to assess the effect of ROS on collagen secretion by aortic and graft SMCs and explore the mechanism involved. METHODS: SMCs isolated from canine aorta or Dacron thoracoabdominal grafts were incubated with 6-anilinoquinoline 5,8-quinone (LY83583), an agent that induces superoxide production. Type I collagen in the conditioned medium was measured by enzyme-linked immunosorbent assay, and superoxide anion production was measured by lucigenin assay. RESULTS: LY83583 stimulated a rapid increase in collagen production by graft SMCs that paralleled the LY83583-induced increase in superoxide production. The increase in both collagen and superoxide was greater in graft SMCs than aortic SMCs. Collagen and superoxide production were inhibited by superoxide scavengers. Nicotinamide adenine dinucleotide phosphate (NADPH) induced significantly more superoxide production by graft SMCs than aortic SMCs, suggesting that the NADPH oxidase system was more active in graft SMCs. NADPH oxidase inhibitors blocked the superoxide and collagen production induced by LY83583. CONCLUSION: In SMCs, the synthetic phenotype is associated with increased NADPH oxidase activity and elevated superoxide production in response to an oxidative stress. Superoxide, in turn, leads to increased collagen production. CLINICAL RELEVANCE: The inflammatory process after prosthetic vascular graft implantation causes oxidative stress that can stimulate collagen production by graft SMCs, contributing to the progression of intimal hyperplasia. The exaggerated response of graft SMCs to oxidative stress offers a potential target for therapeutic interventions. PMID- 16476617 TI - Downregulation of desmuslin in primary vein incompetence. AB - OBJECTIVE: Primary vein incompetence is one of the most common diseases of the peripheral veins, but its pathogenesis is unknown. These veins present obvious congenital defects, and examination of gene expression profiles of the incompetent vein specimens may provide important clues. The aim of this study was to screen for genes affecting the primary vein incompetence phenotype and test the differential expression of certain genes. METHODS: We compared gene expression profiles of valvular areas from incompetent and normal great saphenous veins at the saphenofemoral junctions by fluorescent differential display reverse transcription polymerase chain reaction (FDD RT-PCR). Differentially expressed complimentary DNAs (cDNAs) were confirmed by Northern blotting and semi quantitative RT-PCR. Similarity of the cDNAs sequences to GenBank sequences was determined. Gene expression status was then determined by Western blot analysis and immunohistochemical techniques. RESULTS: There were >30 differentially expressed cDNA bands. Sequence analysis revealed that a cDNA fragment obviously downregulated in incompetent great saphenous vein was a portion of the messenger RNA (mRNA) encoding desmuslin, a newly discovered intermittent filament protein. Northern blotting and semi-quantitative RT-PCR analysis revealed a similar mRNA expression profile of the desmuslin gene in other samples. Western blotting and immunohistochemical techniques localized the desmuslin protein mainly in the cytoplasm of venous smooth muscle cells. The amount of desmuslin was greatly decreased in the smooth muscle cells of incompetent veins. CONCLUSIONS: The expression of many genes is altered in primary vein incompetence. Up- or downregulation of these genes may be involved in the pathogenesis of this disease. Desmuslin expression is downregulated in the abnormal veins. Its effect on the integrity of smooth muscle cells might be related to malformation of the vein wall. Further studies are needed to investigate other differentially expressed cDNAs and the exact role of desmuslin in this disease. CLINICAL RELEVANCE: Primary vein incompetence is a frequent and refractory disease of the peripheral veins. Exploring its pathogenesis may enhance our comprehension and management of this disease. We used reliable techniques to detect disease-related genes and confirmed downregulation of desmuslin in abnormal veins. Alteration of these genes might be used as disease markers or gene therapy targets. PMID- 16476619 TI - Shifting Wallgraft position: case reports and review of the forces affecting Wallgraft positioning. AB - Extensive use of stent-grafts in the iliac arteries is an increasingly common endovascular alternative to an aortobifemoral bypass procedure. The treatment of diffuse bilateral disease is frequently performed by using the technique of paired stent-grafts in the proximal common iliac arteries to effectively re create the aortic-bifurcation. We present three patients treated with this technique who returned with recurrent occlusive disease. During subsequent treatment, the stent-grafts were noted to be in different positions from where they were initially deployed. A discussion of the possible explanations for the shifting positions follows. PMID- 16476618 TI - Infected carotid pseudoaneurysm and carotid-cutaneous fistula as a late complication of carotid artery stenting. AB - Carotid stenosis after neck irradiation is a well-described entity. A 78-year-old man presented with left eye amaurosis fugax 11 years after radical neck dissection and neck irradiation for mucoepidermoid carcinoma. The patient underwent carotid artery stenting for a high-grade stenosis of the left internal carotid artery. Twenty months after the procedure, a pulsatile neck mass developed with intermittent arterial bleeding. After control of the bleeding, he underwent resection of the left carotid bifurcation, including the indwelling stent and reconstruction with a saphenous vein interposition graft. A pectoralis myocutaneous flap was used for wound closure. In this case, localized infection of the stented carotid artery led to mycotic degeneration, pseudoaneurysm formation, and erosion to the skin surface. As carotid artery stenting becomes more widely used, previously unreported late complications associated with this procedure are likely to become apparent, and continued close follow-up is warranted. PMID- 16476620 TI - Percutaneous endovascular treatment of acute sequential systemic emboli. AB - A 65-year-old woman was evaluated for acute right lower-extremity ischemia secondary to embolization from atrial fibrillation and subtherapeutic anticoagulation. Her symptoms quickly evolved in a dynamic fashion to involve multiple vascular beds, including a transient ischemic attack, abdominal pain secondary to mesenteric ischemia, and contralateral lower-extremity ischemia. Synchronous emboli were treated by using mechanical and pharmacologic thrombolysis via an endovascular approach under local anesthesia. This intervention allows prompt evaluation of multiple vascular beds with good short term outcomes and minimal morbidity. PMID- 16476621 TI - Aortic endograft infection due to Pasteurella multocida following a rabbit bite. AB - Abdominal aortic endograft infection is a serious complication after an endovascular abdominal aortic aneurysm repair. Pasteurella multocida, a gram negative bacterium, is a commonly found organism in the mouth flora of many house pets. We report a case of an aortic endograft infection caused by P multocida after a rabbit bite. Successful treatment was performed by extra-anatomic revascularization followed by endograft removal. PMID- 16476622 TI - Successful repair of blunt injury of aortic arch branches in the setting of bovine arch. AB - Intrathoracic great vessels injuries are usually fatal and represent a special surgical challenge. We report a successful surgical repair for blunt disruption of the three aortic arch branches in the setting of bovine arch anatomy. The repair was achieved without the use of cardiopulmonary bypass or arterial shunts. There was no clinical or radiologic evidence of neurologic abnormality after the repair. PMID- 16476623 TI - Use of an endovascular occlusion balloon for control of unremitting venous hemorrhage. AB - This report describes a new approach for management of iliac vein injury. These injuries are often difficult to expose, and the associated hemorrhage further hinders visualization and subsequent repair. In this case, the use of an endovascular balloon from groin access controlled venous hemorrhage and permitted a primary repair of a torn left iliac vein. We believe that this approach is unique in that it uses a compliant, low-pressure balloon, thus preventing further iatrogenic injury in otherwise fragile venous structures and allowing direct access to the tear when exposure in the operative field is limited. PMID- 16476624 TI - Carotid string sign. PMID- 16476625 TI - T-banding: a technique for flow reduction of a hyperfunctioning arteriovenous fistula. AB - A common procedure for hyperfunctioning arteriovenous fistulas is banding, baring the potential for complications such as pre- and poststenotic dilatations and recurrence of high flow caused by slipping of the band itself. We describe a new technique that ensures proper fixation of the graft to prevent anastomotic aneurysms and enlarge the length of stenosis of the shunt vein in order to amplify the effect of flow reduction. This technique was used in 22 patients, with a mean flow reduction of about 50%. The procedure was effective and safe at 1 to 3 months follow-up. PMID- 16476626 TI - Current status of carotid artery stenting. AB - This Clinical Update summarizes the results of larger case series, industry sponsored registries, and randomized trials of carotid artery stenting (CAS). In >20 case series that studied >24,000 patients undergoing CAS, 51% of patients were symptomatic, most procedures (97%) resulted in successful stent deployment, and 30-day stroke rates varied from 1% to 8%, with a trend toward lower rates as experience and embolic protection device (EPD) use increased. In 12 industry sponsored registries (none were published in peer-reviewed journals), 30-day stroke rates varied from 2% to 7%, and 30-day combined adverse events, including stroke, death, and myocardial infarction, were 3% to 9%. More than 12 randomized trials comparing CAS and carotid endarterectomy (CEA) have been initiated since 1998. Results have varied over time, depending on the population studied and the technology used. However, the largest and most recent results of the completed SAPPHIRE trial in high-risk patients undergoing CAS with the use of EPDs demonstrated that CAS is at least not inferior to CEA, with a 1-year combined adverse event rate of 12% for CAS and 20% for CEA (P = .05). Other ongoing trials will address not only whether CAS could be superior to CEA in high-risk patients but also, more importantly, whether CAS is beneficial in other subgroups, such as low-risk and asymptomatic patients. PMID- 16476627 TI - Vascular and endovascular surgeon: the vascular specialist for the 21st century and beyond. PMID- 16476628 TI - A helping hand bitten: an ethical response to medical malpractice suits. PMID- 16476630 TI - Regarding "Value of the duplex waveform at the common femoral artery for diagnosing obstructive aortoiliac disease". PMID- 16476631 TI - Regarding "Prevention of renal failure in patients undergoing thoracoabdominal aortic aneurysm repair". PMID- 16476633 TI - Effect of language on heart attack and stroke awareness among U.S. Hispanics. AB - BACKGROUND: Hispanics with acute heart attack or stroke have longer delay times to hospital arrival, and are thus less likely to benefit from time-dependent reperfusion therapies. Delay time is influenced largely by recognition of warning symptoms and activation of the 911 system. Previous studies suggest poor symptom recognition among racial/ethnic minorities, but very little is known about heart attack and stroke knowledge among Hispanics, or the influence of English language proficiency on cardiovascular emergency awareness. METHODS: Cross-sectional analysis (conducted in 2005) of data from the 2003 Behavioral Risk Factor Surveillance System population survey was limited to states with Spanish-language interviews. Principal study outcomes were the correct identification of five heart attack and five stroke warning symptoms, and intention to call 911 for a suspected event. Subjects included 698 English-speaking Hispanics, 527 Spanish speaking Hispanics, and 24,201 non-Hispanics. RESULTS: Spanish-speaking Hispanics are far less likely to know all heart attack symptoms (7%) than English-speaking Hispanics (23%), non-Hispanic blacks (28%), and non-Hispanic whites (39%) (p<0.001); and far less likely to know all stroke symptoms (18%) than English speaking Hispanics (31%), non-Hispanic blacks (41%), and non-Hispanic whites (50%) (p<0.001). Intention to call 911 did not differ significantly among groups. Spanish-speaking Hispanics remain significantly less likely than all other groups to correctly identify symptoms after adjustment for sociodemographic characteristics, healthcare access, and cardiovascular risk factors (p<0.05). CONCLUSIONS: Lack of English proficiency is strongly associated with lack of heart attack and stroke knowledge among Hispanics. This highlights the need for educational intervention about cardiovascular emergencies targeted to Spanish speaking communities. PMID- 16476634 TI - Obesity and socioeconomic status in children of Tijuana. AB - BACKGROUND: Although obesity has been associated with socioeconomic status among Hispanics living in the United States, little is known about whether a similar association exists among Hispanics living in Mexico, particularly among those living along the U.S.-Mexico border. OBJECTIVE: To determine the prevalence of obesity and its association with socioeconomic status in Mexican schoolchildren attending public and private schools in Tijuana, Baja California. METHODS: Anthropometric measurements and socioeconomic status were assessed in a cross sectional study of 1172 school children, aged 6 to 13 years from 55 schools in Tijuana in 2001-2002. Underweight (body mass index [BMI] for age 5th percentile or lower), risk of overweight (BMI at 85th percentile or higher), and overweight (BMI greater than 95th percentile) were assessed using charts published by the Centers for Disease Control and Prevention. RESULTS: Abnormalities in weight were found in 46.3% of 587 boys and 43.7% of 585 girls in the study. Undernutrition was found in 3.7% of the boys and 3.8% of the girls. The general prevalence of overweight was 23.2% for boys and 21.7% for girls. Children living in low-income neighborhoods had the thickest biceps skinfolds (p<0.01), while children living in moderate-income neighborhoods and attending public schools had the thickest triceps skinfolds (p<0.001). Although boys living in high socioeconomic status neighborhoods were at decreased risk for being overweight, boys and girls attending private schools had a 75% increased risk (odds ratio, 1.75; confidence interval, 1.22-2.52) of being overweight than children attending public schools. CONCLUSIONS: Adiposity varies by type of school and neighborhood socioeconomic status. The biphasic curve in risk for being overweight associated with neighborhood socioeconomic status suggests that Mexican children living along the U.S. border may be experiencing a nutrition transition with respect to an increased risk of obesity and related chronic disease. PMID- 16476635 TI - Colorectal cancer screening among Mexican Americans at a community clinic. AB - BACKGROUND: Mexican Americans tend to under-utilize colorectal cancer (CRC) prevention. Yet little is known about sociocultural factors associated with CRC screening. This study assessed predictors of three primary CRC tests among low income Mexican Americans. METHODS: From May to December 2003, an availability sample of 287 patients, aged 50 to 89 years, who presented for routine care at a community health center near the U.S.-Mexico border completed surveys on CRC knowledge, awareness, attitudes toward screening, logistic barriers, perceptions of health, locus of control, acculturation, whether their doctor discussed CRC screening, and sociodemographics. Participants also reported whether they had ever had a fecal occult blood test, flexible sigmoidoscopy, or colonoscopy. Logistic regression identified predictors of having had these tests. RESULTS: Overall, 41% reported having ever had any of the three tests; 34.1% had a fecal occult blood test; 6.6%, flexible sigmoidoscopy; and 11.8%, colonoscopy. Few respondents reported any clear knowledge about CRC, and only 41% said their doctor had ever discussed screening with them. Yet "doctor discussed screening" was the only consistent screening predictor across tests. CRC knowledge (p=0.006) and insurance coverage (p=0.009) predicted having had a flexible sigmoidoscopy. Perceptions of general poor health also predicted having had a flexible sigmoidoscopy or a colonoscopy (p=0.04). Being employed marginally predicted whether patient had ever had any of the three tests (p=0.05). CONCLUSIONS: Results show that even those in contact with community medical services exhibit low CRC screening rates. They further suggest that interventions focused on clinical settings are an important first step toward CRC prevention in this community. PMID- 16476636 TI - Assessing prevention research impact: a bibliometric analysis. AB - BACKGROUND: This study was undertaken to explore a bibliometric approach to assessing the impact of selected prevention research center (PRC) peer-reviewed publications. METHODS: The 25 eligible PRCs were asked to submit 15 papers that they considered the most important to be published in the decade 1994-2004. Journal articles (n=227) were verified in 2004 and categorized: 73% were research reports, 10% discussion articles, 9% dissemination articles, and 7% review articles. RESULTS: Only 189 articles (83%) were searchable via the Institute of Scientific Information (ISI) Web of Science databases for citation tracking in 2004. These 189 articles were published in 76 distinct journals and subsequently cited 4628 times (range 0 to 1523) in 1013 journals. Articles published before 2001 were cited a median of 14 times each. Publishing journals had a median ISI impact factor of 2.6, and ISI half-life of 7.2. No suitable benchmarks were available for comparison. The PRC influence factor (number of PRCs that considered a journal highly influential) was only weakly correlated with the ISI impact factor and was not correlated with half-life. CONCLUSIONS: Conventional bibliometric analysis to assess the scientific impact of public health prevention research is feasible, but of limited utility because of omissions from ISI's databases, and because citation benchmarks for prevention research have not been established: these problems can and should be addressed. Assessment of impact on public health practice, policy, or on the health of populations, will require more than a bibliometric approach. PMID- 16476637 TI - Impact of smoking cessation aids and mass media among recent quitters. AB - BACKGROUND: Although studies have addressed the effectiveness of conventional smoking aids such as quit-smoking programs and pharmaceutical therapy, few studies have assessed their likely impact on cessation at the population level relative to the impact of mass media anti-tobacco advertisements. METHODS: A random digit dial telephone survey of 6739 Massachusetts residents conducted in 2001-2002 yielded a subsample of 787 individuals who had quit-smoking within the past 2 years. Measures included the types of cessation aids used and perceptions of their helpfulness. Rates of population impact were estimated. Multinomial logistic regression determined the predictors of being helped by conventional aids, by TV advertisements only, or having no help. RESULTS: Analyses conducted in 2004-2005 showed that advertisements were the most frequently mentioned source of help among recent quitters. Older more dependent smokers were most likely to find conventional aids helpful. Younger respondents and those who had remained abstinent for more than 6 months were most likely to report being helped by TV ads. The most helpful ads were those that depicted illness due to smoking or provided inspirational quit tips. CONCLUSIONS: Anti-tobacco media campaigns are a vital component of the National Action Plan for Tobacco Cessation. It is essential that such a campaign be implemented, both to support the National Quit Line and to provide assistance to those smokers who find no other form of aid helpful. PMID- 16476638 TI - Cigarette promotional offers: who takes advantage? AB - BACKGROUND: Promotional offers on cigarettes (e.g., dollar-off, multipack discounts) composed the largest share of tobacco industry marketing expenditures, totaling $8.9 billion, or 72% of the total budget in 2002. Internal industry documents indicate that young adults, potential quitters, and other price sensitive groups are the targets of these marketing tactics. How effective they are in actually reaching these groups in the general population of smokers has not yet been investigated. METHODS: Data were from 4618 current smokers responding to the large, random-digit-dialed population-based 2002 California Tobacco Survey. The characteristics were identified of smokers who reported that they used these offers "every time I see one." RESULTS: Thirty-five percent of smokers used promotional offers every time they saw one. Multivariate analyses identified young adults, women, African Americans, those with higher daily cigarette consumption, and those worried about cigarette costs as more likely to use promotional offers at every opportunity. Smokers most committed to quitting were no more likely to use promotional offers than those with no intention to quit. Cigarette brand was highly correlated with age and race/ethnicity, and therefore was not included in the multivariate analysis. Those who smoked menthol cigarettes and Camels, more often young adults and African Americans, were much more likely than those of other brands to use promotional offers. CONCLUSIONS: With the exception of smokers intending to quit, cigarette promotional offers are effectively reaching most industry-targeted groups. Importantly, young adults, who have the greatest long-term customer potential, are responding. PMID- 16476639 TI - Increasing evidence of immunity to varicella among children in Massachusetts, 1999-2003. AB - BACKGROUND: Experiences with vaccine-preventable diseases have demonstrated the success of school-entry requirements in increasing vaccination coverage and decreasing disease incidence. This study examines the effect of early implementation of daycare and school-entry requirements for varicella vaccination on recorded varicella immunity of preschool and school-aged children in Massachusetts. METHODS: Immunization surveys were conducted in licensed child care centers and schools with kindergarten and/or 7th grades. Evidence of immunity to varicella was defined as having physician verified records of varicella vaccination or disease history from the 1999-2000 through 2003-2004 school years. RESULTS: During the 5-year study period, physician-certified reliable history of varicella disease decreased in each grade level while vaccination coverage increased. The increase in the number of children in each grade level receiving varicella vaccine led to an increase in the overall percentage of children with evidence of immunity to varicella: 85% to 97% for children aged 2 years or more in child care, 93% to 98% for children in kindergarten, and 88% to 92% for children in 7th grade. CONCLUSIONS: The implementation of daycare and school-entry requirements for varicella vaccination within 4 years of the start of the varicella vaccination program in Massachusetts was associated with high levels of vaccination coverage in the cohorts of children targeted by the requirements. Although evidence of immunity from varicella disease decreased during the study period, the increase in varicella vaccination coverage compensated for the decline in disease history, resulting in a higher proportion of young children with evidence of immunity to varicella. PMID- 16476640 TI - Effect of 5-year community intervention Hartslag Limburg on cardiovascular risk factors. AB - BACKGROUND: A widely advocated strategy in public health is community-based health promotion. The aim of this study was to investigate the net effect of a cardiovascular disease prevention program (Hartslag Limburg) on cardiovascular risk factors after 5 years of intervention. DESIGN: Cohort study comparing 5-year mean change in risk factors between the intervention and reference area. The statistical analyses for the study were performed in 2005. SETTING/PARTICIPANTS: In 1998, 3000 subjects (aged 25 to 70) from the intervention area and 895 subjects from a reference area participated in the baseline measurement. Of these, 2414 intervention subjects and 758 reference subjects completed the follow up measurement in 2003. INTERVENTION: Hartslag Limburg is an integrative community-based cardiovascular disease prevention program promoting a healthy lifestyle. MAIN OUTCOME MEASURES: Body mass index (BMI), waist circumference, blood pressure, serum glucose (nonfasting), and serum total and high-density lipoprotein (HDL) cholesterol. RESULTS: During the 5-year follow-up, risk factors changed unfavorably in the reference group, whereas changes were less pronounced or absent in the intervention group. The adjusted difference in mean change in risk factors between intervention and reference group was significant (p<0.05) for BMI: -0.36 kg/m(2) in men and -0.25 kg/m(2) in women; waist circumference 2.9 cm in men and -2.1 cm in women; systolic blood pressure: -7.8 mmHg in men and -5.5 mmHg in women; total cholesterol 0.11 mmol/L in women and finally serum glucose -0.23 mmol/L in women. CONCLUSIONS: Hartslag Limburg succeeded in reducing-and in some cases, preventing-age- and time-related increase in BMI, waist circumference, blood pressure, and, in women, nonfasting glucose concentration. PMID- 16476641 TI - Evaluation of a community health promotion resource for primary care practices. AB - BACKGROUND: Primary care physicians' ability to provide effective health behavior change advice might be leveraged by linking to available community resources. This study evaluates tools to facilitate such a link. DESIGN: A mixed methods longitudinal pre-post-test study was conducted in 2004. SETTING/ PARTICIPANTS: Seven primary care practices in northeast Ohio and two longitudinal cohorts of patients (n=784). INTERVENTION: The practice-tailored intervention included two main components: (1) a web-based health behavior change resource including a database of community programs and patient education materials, and (2) a health behavior prescription pad. MAIN OUTCOME MEASURES: Rates of discussion of diet, exercise, weight management, and smoking cessation; and patient change in motivation to modify behaviors at 8 weeks post-visit. Qualitative field notes about practice routines, culture, and implementation efforts were analyzed to enhance understanding of the practice change process. RESULTS: The post intervention cohort reported higher rates of discussion of diet (25.7% vs 20.2%), exercise (27.8% vs 16.9%), and weight management (23.2% vs 16.3%), and greater referral to patient education materials (24.2% vs 21.6%) and community programs for health behavior change (16.0% vs 13.6%) (p<0.05). No difference in change in patient motivation to modify behavior was observed between the pre- and post intervention cohorts. Evaluation of the qualitative field notes suggests that for six of seven practices, the intervention was not appreciably implemented. CONCLUSIONS: The intervention tools show promise for increasing clinician discussion of health behaviors, information referral, and referral to community resources. Additional work to increase practice adoption of the tools is required to understand their potential to affect patient motivation to change behavior. PMID- 16476642 TI - Epidemiology of gastroenteritis on cruise ships, 2001-2004. AB - BACKGROUND: The incidence of diarrheal disease among cruise ship passengers declined from 29.2 cases per 100,000 passenger days in 1990 to 16.3 per 100,000 passenger days in 2000. In 2002, the Vessel Sanitation Program of the Centers for Disease Control and Prevention reported 29 outbreaks (3% or more passengers ill) of acute gastroenteritis on cruise ships, an increase from 3 the previous year. This analysis of gastroenteritis on cruise ships, conducted in 2005, details the increase in outbreak incidence rates during 2001 through 2004. METHODS: Using Gastrointestinal Illness Surveillance System data, investigators evaluated incidence rates of gastroenteritis on cruise ships calling on U.S. ports, carrying 13 or more passengers, by cruise length and reporting region during the study period. The investigators also evaluated the association between inspection scores, and gastroenteritis incidence and the frequency of outbreaks in 2001 through 2004. RESULTS: During the study period, the background and outbreak associated incidence rates of passengers with acute gastroenteritis per cruise were 25.6 and 85, respectively. Acute gastroenteritis outbreaks per 1000 cruises increased overall from 0.65 in 2001 to 5.46 in 2004; outbreaks increased from 2 in 2001 to a median of 15 per year in 2002-2004. Median ship inspection scores remained relatively constant during the study period (median 95 on a 100-point scale), and were not significantly associated with either gastroenteritis incidence rates (risk ratio, 1.00; 95% confidence interval, 0.98-1.02) or outbreak frequency (Spearman's coefficient, 0.01, p=0.84). CONCLUSIONS: Despite good performance on environment health sanitation inspections by cruise ships, the expectation of passenger cases of gastroenteritis on an average 7-day cruise increased from two cases during 1990-2000 to three cases during the study period. This increase, likely attributable to noroviruses, highlights the inability of environmental programs to fully predict and prevent risk factors common to person to-person and fomite spread of disease. PMID- 16476643 TI - Healthcare workers who elected not to receive smallpox vaccination. AB - BACKGROUND: The goal of the National Smallpox Vaccination Program was to vaccinate a cadre of healthcare workers and first responders who could care for smallpox patients in the event of an attack. METHODS: Using a convenience sample of health departments (n=49) and hospitals (n=60) in five states, we conducted a telephone interview between July 2003 and April 2004 of healthcare workers and first responders who chose not to receive smallpox vaccination. (Data were analyzed in 2004 and 2005.) RESULTS: The response rate was 63%. Of 1895 respondents, 723 (38.2%) reported having a contraindication, 280 (14.8%) reported being contraindicated because of a household member's condition, and 892 (47.0%) reported having no contraindication to smallpox vaccination. Among respondents with no contraindication, the leading reasons for nonvaccination were concerns about side effects (20.6%) and not feeling that the risk of outbreak was high enough (19.5%). More than half (54.8%) were somewhat or very concerned about having an adverse reaction to the vaccine; Hispanics, blacks, and Asians were significantly more likely than whites to be somewhat or very concerned about side effects. Less than one fifth (17.9%) reported that there was a policy to financially compensate employees who developed side effects from vaccination, and 40.7% reported that there was a policy to provide liability coverage to employees who transmitted vaccinia to a patient. CONCLUSIONS: Many people who chose not to receive smallpox vaccine perceived their personal risk-benefit balance as not favoring vaccination. The success of future smallpox vaccination efforts or vaccination against other bioterrorist health threats depends on addressing potential barriers to participation including compensation and liability issues, in addition to clearly communicating risks and benefits. PMID- 16476644 TI - Availability of nutrition information from chain restaurants in the United States. AB - BACKGROUND: Although obesity and poor dietary habits are complex multifactorial problems, away-from-home food has been identified as one likely and important contributor. Restaurants provide a growing and substantial portion of the average American's diet, yet the Nutrition Labeling and Education Act (NLEA), which went into effect in 1994, explicitly exempts restaurants from most labeling requirements. Thus, this study examined the availability of nutrition information from the largest chain restaurants in the United States. METHODS: Between January and August 2004, we surveyed the 300 largest chain restaurants by telephone, e mail, or examining company websites (response rate was 96%). The top chains, as ranked by revenue, were selected based on 2002 ratings in Restaurants and Institutions. RESULTS: Fifty-four percent of the 287 largest chain restaurants made some nutrition information available. Forty-four percent had nutrition information for the majority of their standard menu items. We found no significant differences in the availability of nutrition information based on the size of the restaurant chain. Of those restaurants with nutrition information, 86% provided information on the company website. CONCLUSIONS: The number of restaurants providing nutrition information has increased over the last 10 years. However, making informed and healthful food choices is hampered by the absence of nutrition information at many restaurants. Given the growing and significant role that away-from-home foods play in Americans' diets, the Surgeon General and the National Academies' Institute of Medicine recommend that nutrition information be available to customers at restaurants, and state legislatures and the U.S. Congress are beginning to address the issue. PMID- 16476646 TI - To err is human... sometimes. PMID- 16476645 TI - Health burden and funding at the Centers for Disease Control and Prevention. AB - BACKGROUND: The relationship between domestic funding for selected conditions to the Centers for Disease Control and Prevention (CDC) and the burden of disease and disability in the United States was assessed systematically. METHODS: Using mortality, years of potential life lost (YPLLs), disability-adjusted life years (DALYs), hospital days, hospital discharges, and direct medical costs of conditions, 34 high-burden conditions addressed by CDC programs were identified, and information was collected about the funds spent on each by CDC during fiscal year (FY) 2003. The 34 conditions were grouped into 15 categorical areas, and the relationship between budget and burden was analyzed using correlation and regression methods for each of the categorical areas and for each measure of burden. RESULTS: Of CDC's total FY 2003 budget of $6.9 billion, 62% ($4.3 billion) of funding was allocated to one of the 34 conditions studied. A positive relationship between budget and burden was identified for all measures of burden, although the correlations varied for the different conditions. CONCLUSIONS: Although examination of the relationship of CDC's budget to burden measures provides insight into the agency's portfolio of investments, this exercise also highlights a number of limitations with this approach and the currently available burden measures. Assessment of key public health functions such as emergency preparedness and the collection of vital statistics require development of metrics different from the burden measures used in this analysis. Investment in the development of such metrics warrants consideration. PMID- 16476647 TI - Web-based learning: an alternative to classroom learning. PMID- 16476648 TI - Driver contrast sensitivity and reaction times as measured through a salt-covered windshield. AB - BACKGROUND: The objective of this study was to use contrast sensitivity measurements to determine the effect of windshield salt film on driver vision. This effect simulates road salt spray that dries in place on a vehicle windshield during winter driving conditions. METHODS: Ten emmetropic subjects were tested to assess contrast sensitivity functions through windshields that were clear compared with the same stimuli as viewed through windshields coated with salt film. Achromatic stimuli were generated by a VisionWorks system and were presented at 4 spatial frequencies (0.5, 3, 10, and 20 cycles per degree). RESULTS: A significant reduction in contrast sensitivity through the salt film was found at all spatial frequencies. In addition, reaction time to detect the stimuli was also found to be increased significantly at 2 of the 4 spatial frequencies through the saline-covered windshield. CONCLUSION: The results of this study indicate that windshield salt film reduces the ability to detect high and low contrast objects in a simulated driving environment as well as increases the subjects' reaction times. PMID- 16476649 TI - Early ocular prosthesis insertion improves quality of life after enucleation. AB - BACKGROUND: The aim of this study was to evaluate early insertion of ocular prostheses for aesthetic rehabilitation after enucleation and review current methods of enucleation surgery. METHODS: Twelve patients with large intraocular malignancies were treated by enucleation with implant. At the end of surgery, a medium-size conformer was inserted onto the closed conjunctival wound (just before applying a pressure dressing). The pressure dressing was removed 3 to 5 days after surgery. Immediately after bandage removal, the patients saw themselves in a mirror and were offered placement of a relatively small ocular prosthesis. Patients were asked if they preferred their aesthetic appearance with or without the prosthesis. RESULTS: Eight patients (67%) tolerated introduction of the prosthesis at the time of removal of their pressure dressing (3 to 5 days after enucleation surgery). Because of discomfort, 4 were able to receive their prostheses at their second postoperative visit (day 12 or 13). The manipulation at prosthesis insertion and subsequent wearing was found to have no effect on wound closure, final prosthetic fittings, or movement of the permanent prosthesis. All patients were pleased with the cosmetic benefit afforded by early introduction of the prosthesis and preferred their appearance to that of their eye with the conformer. CONCLUSIONS: Early replacement of the conformer by an ocular prosthesis allows for cosmetic rehabilitation and improved quality of life. PMID- 16476650 TI - Acute onset of halos and glare: bilateral corneal epithelial edema with cystic eruptions--atypical presentation of amiodarone keratopathy. AB - BACKGROUND: Amiodarone (Cordarone; Wyeth, Ayerst, New York) is a potassium channel blocking antiarrythmal medication indicated for recurrent ventricular fibrillation and recurrent hemodynamically unstable ventricular tachycardia. Chemically, it is classified as an iodinated benzofuran derivate antiarrythmal drug not chemically related to any other available antiarrhythmic drug. Documented side effects of amiodarone include neurologic, gastrointestinal, dermatologic, cardiovascular, and ophthalmic. The ophthalmic abnormalities include optic neuropathy, optic neuritis, papilledema, corneal deposits, photosensitivity, lens opacities, and macular degeneration. Corneal microdeposits are seen in virtually all patients who receive amiodarone for more than 6 months. Corneal microdeposits result secondary to the secretion of amiodarone by the lacrimal gland with accumulation on, and absorption by, the corneal epithelium. Approximately 10% of these patients become symptomatic with glare and halos; however, that alone is usually not enough to precipitate intervention. CASE REPORT: A 69-year-old woman presented to our office with a 2-week history of halos and glare in both eyes (OU). Her ocular history was significant for occasional contact lens wear. Her systemic history was significant for atrial fibrillation, for which she was taking amiodarone daily for the last 6 years. Six weeks before the onset of her chief complaint, her daily amiodarone dosage was increased from 100 mg to 300 mg. Ophthalmic examination found decreased visual acuities OU, amiodarone keratopathy (subepithelial verticillata), and diffuse corneal epithelial edema with diffuse sodium fluorescein staining OU. The corneal compromise was treated in 2 ways: the source medication was discontinued, and the ocular signs were medicated with a prophylactic topical antibiotic along with supportive preparations (tears/lubricants) and monitored over 2 weeks until full elimination of the pathologic signs and their symptoms. Even after complete resolution of the acute keratitis, the infiltrative keratopathy persisted along with the initial complaints of halos and glare. The supportive treatments were maintained over the course of 2 months until full recovery. CONCLUSION: By exclusion, it was determined that the subepithelial depositions and cystic formations were secondary to an acute amiodarone dosage increase by a new practitioner. Prompt communication resulted in the physician discontinuing that medicine. PMID- 16476651 TI - Three presentations of monocular vision loss. AB - BACKGROUND: Carotid artery disease is estimated to affect 30% of persons older than 50. Risk factors include hypertension, cigarette smoking, hyperlipidemia and diabetes mellitus. Symptoms ascribed to carotid artery lesions with stenosis of the artery or plaque formation include monocular vision loss and transient ischemic attacks. Patients can present with transient monocular vision loss as their initial symptom. CASE REPORTS: Three patients from a geriatric clinic in Wilmington, Delaware presented with different complaints of vision loss with similar overall outcomes. Patient A was an 87-year-old woman who presented with dimming of vision on extreme left head turn. Dilated fundus examination found a retinal arterial emboli in the left eye (O.S.). Carotid duplex examination found 50% to 79% left internal carotid stenosis with no hemodynamic stenosis of the right internal carotid. Patient B was a 78-year-old woman who presented with a right superior altitudinal defect and transient vision loss in the right eye. Dilated fundus examination found retinal arterial emboli in the right eye (O.D.). Carotid duplex examination found 50% to 79% carotid stenosis in both the left and right internal carotids. Patient C was an 84-year-old man who complained of a superior altitudinal visual field defect O.D. Dilated fundus examination found a retinal arterial emboli O.D. Carotid duplex results showed calcified atherosclerotic plaques present at the level of the common carotid artery bifurcations bilaterally, with 50% to 70% narrowing of the right internal carotid artery with no significant narrowing of the left internal carotid artery. CONCLUSIONS: These 3 presentations show that in patients older than 50 who present with chief complaints of monocular vision loss, a differential diagnosis of carotid artery disease must be considered. Patients who exhibit retinal arterial emboli are at increased risk for stroke and vascular death. Appropriate measures for confirming a diagnosis include duplex ultrasound imaging, magnetic resonance angiography (MRA), and carotid angiography. Surgical techniques such as carotid angioplasty and carotid endarterectomy may be recommended. PMID- 16476652 TI - Enhanced visual experiences and seeing hardware for reduced vision: a pilot study. AB - BACKGROUND: Persons with vision loss may have limited ability to experience visual arts and difficulty navigating unfamiliar environments. The Center for Advanced Visual Studies at Massachusetts Institute of Technology developed a Retinal Imaging Machine Vision System (RIMVS) "seeing machine" to project "word images" onto the retina and permit navigation through virtual architectural environments. METHODS: Ten subjects with visual acuity < or = 20/70 in the better seeing eye were recruited. Subjects viewed word-images that were projected on the retina of the better-seeing eye and asked to interpret the images. Subsequently, they were asked to recall as many images as possible. Participants also practiced navigation through a computer-modeled building that was projected onto the retina. A survey elicited subject response to the RIMVS. RESULTS: Six subjects correctly interpreted all 10 word-images; no subject interpreted fewer than 8 correctly. Seven subjects felt that the word images afforded easy recall, and 7 subjects responded that the technology might assist subsequent navigation in unfamiliar environments. CONCLUSIONS: Responses reflect a general interest and potential with the RIMVS. Further studies are warranted to explore the value of the RIMVS as a tool to enable visually compromised persons to experience and appreciate visual arts and as a navigation aid. PMID- 16476653 TI - Effect of adrenaline on transmesothelial resistance of isolated sheep pleura. AB - The effect of adrenaline on the transmesothelial resistance (RTM) of sheep's visceral and parietal pleura was studied using the Ussing chamber technique. Basal transmesothelial resistance of visceral pleura was found to be 20.71 +/- 0.31 Omega cm2, whereas that of parietal pleura was found to be 19.53 +/- 0.34 Omega cm2. Immediately after the addition of adrenaline (10(-7) M) both apically and basolaterally on the visceral and parietal pleura, these values were significantly increased (P < 0.05). Addition of the nonselective beta-receptor blocker, propranolol (10(-5) M), suppressed this effect in both visceral and parietal pleura, while addition of the nonselective alpha-receptor blocker, phentolamine (10(-5) M), partly suppressed the above-mentioned increase in the parietal pleura. In conclusion, our results show that adrenaline has a rapid effect on both pleurae. This rapid effect is mediated by the stimulation of beta adrenergic receptors in the case of visceral pleura, while in the case of parietal pleura this effect seems to be due to a stimulation of alpha- and beta adrenergic receptors. On the visceral pleura the effect of adrenaline vanishes after some minutes and on the parietal this effect is more permanent than the visceral's one, suggesting differences in the distribution of the adrenergic receptors between the visceral and parietal pleura. PMID- 16476654 TI - Changes in peak expiratory flow and respiratory strength during the menstrual cycle. AB - This study evaluated the spirometry and respiratory static pressures in 17 young women, twice a week for three successive ovulatory menstrual cycles to determine if such variables changed across the menstrual, follicular, periovulatory, early to-mid luteal and late luteal phases. The factors phases of menstrual cycle and individual cycles had no significant effect on the spirometry variables except for peak expiratory flow (PEF) and respiratory static pressures. Significant weak positive correlations were found between the progesterone:estradiol ratio and PEF and between estrogen and tidal volume (r = 0.37), inspiratory time (r = 0.22), expiratory time (r = 0.19), maximal inspiratory pressure (r = 0.25) and maximal expiratory pressure (r = 0.20) and for progesterone and maximal inspiratory pressure (r = 0.32) during the early-to-mid luteal phase. Although most parameters of the spirometry results did not change during the menstrual cycle, the correlations observed between sexual hormones and respiratory control variables suggest a positive influence of sexual female hormones controlling the thoracic pump muscles in the luteal phase. PMID- 16476655 TI - The surface activity of pulmonary surfactant from diving mammals. AB - Pinnipeds (seals and sea lions) have developed a specialised respiratory system to cope with living in a marine environment. They have a highly reinforced lung that can completely collapse and reinflate during diving without any apparent side effects. These animals may also have a specialised surfactant system to augment the morphological adaptations. The surface activity of surfactant from four species of pinniped (California sea lion, Northern elephant seal, Northern fur seal and Ringed seal) was measured using a captive bubble surfactometer (CBS), and compared to two terrestrial species (sheep and cow). The surfactant of Northern elephant seal, Northern fur seal and Ringed seal was unable to reduce surface tension (gamma) to normal levels after 5 min adsorption (61.2, 36.7, and 46.2 +/- 1.7 mN/m, respectively), but California sea lion was able to reach the levels of the cow and sheep (23.4 mN/m for California sea lion, 21.6 +/- 0.3 and 23.0 +/- 1.5 mN/m for cow and sheep, respectively). All pinnipeds were also unable to obtain the very low gamma(min) achieved by cow (1.4 +/- 0.1 mN/m) and sheep (1.5 +/- 0.4 mN/m). These results suggest that reducing surface tension to very low values is not the primary function of surfactant in pinnipeds as it is in terrestrial mammals, but that an anti-adhesive surfactant is more important to enable the lungs to reopen following collapse during deep diving. PMID- 16476656 TI - Sexual dimorphism of human ribs. AB - The volume of the rib cage is about 10% smaller in females than in males having the same height although the reason for this is presently unclear. The cranio caudal inclination of ribs is greater in females than males but the length of ribs has not previously been compared between the sexes. In 23 males and 23 females studied at necropsy, body length, the length of the upper and lower limbs and the length of the thoracic spine were all smaller in females but the ratios of upper and lower limb lengths to body length and of thoracic spine length to body length were not different. By contrast, the lengths of the third, sixth and ninth ribs were not significantly different between males and females and the ratios of rib length to body length were all significantly greater in females. We conclude that in females the ribs grow longer in relation to the axial skeleton than in males. PMID- 16476657 TI - Maintaining your youthful spontaneity: microcircuit homeostasis in the embryonic spinal cord. AB - Many developing networks generate spontaneous network activity (SNA) that plays an important role in setting up functional circuitry, but how the proper level and pattern of SNA is itself maintained has not been clear. In this issue of Neuron, Gonzalez-Islas and Wenner show that SNA in the intact embryo regulates itself through a set of adaptive homeostatic plasticity mechanisms. PMID- 16476658 TI - A new peptide input to learning and addiction. AB - In this issue of Neuron, Borgland et al. report that the arousal-associated peptide orexin enhances LTP-like changes in glutamatergic excitability of ventral tegmental dopamine neurons. This parallels a similar effect of corticotropin releasing factor and suggests a form of neuroadaptation that increases the likelihood of addiction relapse. PMID- 16476659 TI - Dopamine receptor excess and mouse madness. AB - The dopamine hypothesis of schizophrenia is based on evidence that the major antipsychotic drugs act by blocking dopamine D2 receptors and that dopamine releasing drugs worsen symptoms. In this issue of Neuron, Kellendonk et al. report an elegant conditional transgenic mouse overexpressing dopamine D2 receptors selectively in the striatum. Strikingly, these animals display selective cognitive impairment typically associated with frontal cortical defects and abnormal dopamine markers in the prefrontal cortex, suggesting that striatal dopamine receptors can influence cortical dopamine function. PMID- 16476660 TI - Bone marrow-derived microglia play a critical role in restricting senile plaque formation in Alzheimer's disease. AB - Microglia are the immune cells of the brain. Here we show a massive infiltration of highly ramified and elongated microglia within the core of amyloid plaques in transgenic mouse models of Alzheimer's disease (AD). Many of these cells originate from the bone marrow, and the beta-amyloid-40 and -42 isoforms are able to trigger this chemoattraction. These newly recruited cells also exhibit a specific immune reaction to both exogenous and endogenous beta-amyloid in the brain. Creation of a new AD transgenic mouse that expresses the thymidine kinase protein under the control of the CD11b promoter allowed us to show that blood derived microglia and not their resident counterparts have the ability to eliminate amyloid deposits by a cell-specific phagocytic mechanism. These bone marrow-derived microglia are thus very efficient in restricting amyloid deposits. Therapeutic strategies aiming to improve their recruitment could potentially lead to a new powerful tool for the elimination of toxic senile plaques. PMID- 16476661 TI - The zinc finger transcription factor Sp8 regulates the generation and diversity of olfactory bulb interneurons. AB - The molecular mechanisms that regulate the production and diversity of olfactory bulb interneurons remain poorly understood. With the exception of the GABAergic/dopaminergic subtype in the glomerular layer, no information exists concerning the generation of the other subtypes. Here we show that the recently identified zinc finger transcription factor Sp8 is expressed in neurogenic regions, which give rise to olfactory bulb interneurons at embryonic and postnatal time points and remains expressed in the calretinin-expressing and GABAergic/nondopaminergic interneurons of the glomerular layer. Conditional inactivation of Sp8 in the embryonic ventral telencephalon reveals a requirement for the normal generation of these interneuron subtypes. Sp8 conditional mutants exhibit an increase in cell death within the lateral ganglionic eminence and rostral migratory stream. Moreover, mutant neuroblasts/interneurons are misspecified and display abnormal migration patterns in the olfactory bulb, indicating that Sp8 contributes to olfactory bulb interneuron diversity by regulating the survival, migration, and molecular specification of neuroblasts/interneurons. PMID- 16476662 TI - The HSPGs Syndecan and Dallylike bind the receptor phosphatase LAR and exert distinct effects on synaptic development. AB - The formation and plasticity of synaptic connections rely on regulatory interactions between pre- and postsynaptic cells. We show that the Drosophila heparan sulfate proteoglycans (HSPGs) Syndecan (Sdc) and Dallylike (Dlp) are synaptic proteins necessary to control distinct aspects of synaptic biology. Sdc promotes the growth of presynaptic terminals, whereas Dlp regulates active zone form and function. Both Sdc and Dlp bind at high affinity to the protein tyrosine phosphatase LAR, a conserved receptor that controls both NMJ growth and active zone morphogenesis. These data and double mutant assays showing a requirement of LAR for actions of both HSPGs lead to a model in which presynaptic LAR is under complex control, with Sdc promoting and Dlp inhibiting LAR in order to control synapse morphogenesis and function. PMID- 16476663 TI - lazaro encodes a lipid phosphate phosphohydrolase that regulates phosphatidylinositol turnover during Drosophila phototransduction. AB - An essential step in Drosophila phototransduction is the hydrolysis of phosphatidylinositol 4,5 bisphosphate PI(4,5)P2 by phospholipase Cbeta (PLCbeta) to generate a second messenger that opens the light-activated channels TRP and TRPL. Although the identity of this messenger remains unknown, recent evidence has implicated diacylglycerol kinase (DGK), encoded by rdgA, as a key enzyme that regulates its levels, mediating both amplification and response termination. In this study, we demonstrate that lazaro (laza) encodes a lipid phosphate phosphohydrolase (LPP) that functions during phototransduction. We demonstrate that the synergistic activity of laza and rdgA regulates response termination during phototransduction. Analysis of retinal phospholipids revealed a reduction in phosphatidic acid (PA) levels and an associated reduction in phosphatidylinositol (PI) levels. Together our results demonstrate the contribution of PI depletion to the rdgA phenotype and provide evidence that depletion of PI and its metabolites might be a key signal for TRP channel activation in vivo. PMID- 16476665 TI - Spontaneous network activity in the embryonic spinal cord regulates AMPAergic and GABAergic synaptic strength. AB - Spontaneous network activity (SNA) has been described in most developing circuits, including the spinal cord, retina, and hippocampus. Despite the widespread nature of this developmental phenomenon, its role in network maturation is poorly understood. We reduced SNA in the intact embryo and found compensatory increases in synaptic strength of spinal motoneuron inputs. AMPAergic miniature postsynaptic current (mPSC) amplitude and frequency increased following the reduction of activity. Interestingly, excitatory GABAergic mPSCs also increase in amplitude through a process of synaptic scaling. Finally, the normal modulation of GABAergic mPSC amplitude was accelerated. Together, these compensatory responses appear to increase the excitability of the cord and could act to maintain appropriate SNA levels, thus demonstrating a distinct functional role for synaptic homeostasis. Because spontaneous network activity can regulate AMPAergic and GABAergic synaptic strength during development, SNA is likely to play an important role in a coordinated maturation of excitatory and inhibitory synaptic strength. PMID- 16476664 TI - A preformed complex of postsynaptic proteins is involved in excitatory synapse development. AB - Nonsynaptic clusters of postsynaptic proteins have been documented; however, their role remains elusive. We monitored the trafficking of several candidate proteins implicated in synaptogenesis, when nonsynaptic clusters of scaffold proteins are most abundant. We find a protein complex consisting of two populations that differ in their content, mobility, and involvement in synapse formation. One subpopulation is mobile and relies on actin transport for delivery to nascent and existing synapses. These mobile clusters contain the scaffolding proteins PSD-95, GKAP, and Shank. A proportion of mobile clusters that exhibits slow movement and travels short distances contains neuroligin-1. The second group consists of stationary nonsynaptic scaffold complexes that mainly contain neuroligin-1, can recruit synaptophysin-containing axonal transport vesicles, and are readily transformed to functional presynaptic contacts that recycle the vital dye FM 4-64. These results postulate a mechanism whereby preformed scaffold protein complexes serve as predetermined postsynaptic hotspots for establishment of new functional excitatory synapses. PMID- 16476666 TI - A "sample-and-hold" pulse-counting integrator as a mechanism for graded memory underlying sensorimotor adaptation. AB - The mechanisms behind the induction of cellular correlates of memory by sensory input and their contribution to meaningful behavioral changes are largely unknown. We previously reported a graded memory in the form of sensorimotor adaptation in the electromotor output of electric fish. Here we show that the mechanism for this adaptation is a synaptically induced long-lasting shift in intrinsic neuronal excitability. This mechanism rapidly integrates hundreds of spikes in a second, or gradually integrates the same number of spikes delivered over tens of minutes. Thus, this mechanism appears immune to frequency-dependent fluctuations in input and operates as a simple pulse counter over a wide range of time scales, enabling it to transduce graded sensory information into a graded memory and a corresponding change in the behavioral output. This adaptation is based on an NMDA receptor-mediated change in intrinsic excitability of the postsynaptic neurons involving the Ca2+-dependent activation of TRP channels. PMID- 16476667 TI - Orexin A in the VTA is critical for the induction of synaptic plasticity and behavioral sensitization to cocaine. AB - Dopamine neurons in the ventral tegmental area (VTA) represent a critical site of synaptic plasticity induced by addictive drugs. Orexin/hypocretin-containing neurons in the lateral hypothalamus project to the VTA, and behavioral studies have suggested that orexin neurons play an important role in motivation, feeding, and adaptive behaviors. However, the role of orexin signaling in neural plasticity is poorly understood. The present study shows that in vitro application of orexin A induces potentiation of N-methyl-D-aspartate receptor (NMDAR)-mediated neurotransmission via a PLC/PKC-dependent insertion of NMDARs in VTA dopamine neuron synapses. Furthermore, in vivo administration of an orexin 1 receptor antagonist blocks locomotor sensitization to cocaine and occludes cocaine-induced potentiation of excitatory currents in VTA dopamine neurons. These results provide in vitro and in vivo evidence for a critical role of orexin signaling in the VTA in neural plasticity relevant to addiction. PMID- 16476669 TI - The dynamics of hippocampal activation during encoding of overlapping sequences. AB - Sequence disambiguation, the process by which overlapping sequences are kept separate, has been proposed to underlie a wide range of memory capacities supported by the hippocampus, including episodic memory and spatial navigation. We used functional magnetic resonance imaging (fMRI) to explore the dynamic pattern of hippocampal activation during the encoding of sequences of faces. Activation in right posterior hippocampus, only during the encoding of overlapping sequences but not nonoverlapping sequences, was found to correlate robustly with a subject-specific behavioral index of sequence learning. Moreover, our data indicate that hippocampal activation in response to elements common to both sequences in the overlapping sequence pair, may be particularly important for accurate sequence encoding and retrieval. Together, these findings support the conclusion that the human hippocampus is involved in the earliest stage of sequence disambiguation, when memory representations are in the process of being created, and provide empirical support for contemporary computational models of hippocampal function. PMID- 16476668 TI - Transient and selective overexpression of dopamine D2 receptors in the striatum causes persistent abnormalities in prefrontal cortex functioning. AB - Increased activity of D2 receptors (D2Rs) in the striatum has been linked to the pathophysiology of schizophrenia. To determine directly the behavioral and physiological consequences of increased D2R function in the striatum, we generated mice with reversibly increased levels of D2Rs restricted to the striatum. D2 transgenic mice exhibit selective cognitive impairments in working memory tasks and behavioral flexibility without more general cognitive deficits. The deficit in the working memory task persists even after the transgene has been switched off, indicating that it results not from continued overexpression of D2Rs but from excess expression during development. To determine the effects that may mediate the observed cognitive deficits, we analyzed the prefrontal cortex, the brain structure mainly associated with working memory. We found that D2R overexpression in the striatum impacts dopamine levels, rates of dopamine turnover, and activation of D1 receptors in the prefrontal cortex, measures that are critical for working memory. PMID- 16476670 TI - Task and content modulate amygdala-hippocampal connectivity in emotional retrieval. AB - The ability to remember emotional events is crucial for adapting to biologically and socially significant situations. Little is known, however, about the nature of the neural interactions supporting the integration of mnemonic and emotional information. Using fMRI and dynamic models of effective connectivity, we examined regional neural activity and specific interactions between brain regions during a contextual memory retrieval task. We independently manipulated emotional context and relevance of retrieved emotional information to task demands. We show that retrieval of emotionally valenced contextual information is associated with enhanced connectivity from hippocampus to amygdala, structures crucially involved with encoding of emotional events. When retrieval of emotional information is relevant to current behavior, amygdala-hippocampal connectivity increases bidirectionally, under modulatory influences from orbitofrontal cortex, a region implicated in representation of affective value and behavioral guidance. Our findings demonstrate that both memory content and behavioral context impact upon large scale neuronal dynamics underlying emotional retrieval. PMID- 16476671 TI - Epidemiology and clinical characteristics of respiratory syncytial virus infections in Jordan. AB - This study was carried out to determine the prevalence, seasonal distribution of RSV, the signs and symptoms associated with it in Jordan. A total of 200 nasopharyngeal aspirates were obtained from hospitalized children (below 2 years old). RSV was detected in 12.5% of patients using direct immunofluorescence technique. Most infections were associated with bronchilolitis, and higher rates of hypoxemia, retractions, tachypnea, hyperinflation and interstitial infiltrates in 1 to 3 months old children. RSV showed a clear temporal periodicity. The epidemic began in December and disappeared in March with a peak of incidence during February 2003 and January 2004. The seasonal distribution showed a significant correlation with temperature, rainfall and relative humidity. This study provides further information on RSV epidemiology which could help in planning of prevention and control programs in Jordan, distinguishing RSV infections on the basis of the clinical picture and considering RSV between December and March each year. PMID- 16476673 TI - Fetal nuchal translucency scan and early prenatal diagnosis of chromosomal abnormalities by rapid aneuploidy screening: observational study. AB - OBJECTIVE: To investigate an approach for the analysis of samples obtained in screening for trisomy 21 that retains the advantages of quantitative fluorescent polymerase chain reaction (qf-PCR) over full karyotyping and maximises the detection of clinically significant abnormalities. DESIGN: Observational study. SETTING: Tertiary referral centre. SUBJECTS: 17,446 pregnancies, from which chorionic villous samples had been taken after assessment of risk for trisomy 21 by measurement of fetal nuchal translucency (NT) thickness at 11 to 13(+6) weeks of gestation. INTERVENTIONS: Analysis of chorionic villous samples by full karyotyping and by qf-PCR for chromosomes 13, 18, 21, X, and Y. MAIN OUTCOME MEASURE: Detection of clinically significant chromosomal abnormalities. RESULTS: The fetal karyotype was normal in 15,548 (89.1%) cases and abnormal in 1898 (10.9%) cases, including 1722 with a likely clinically significant adverse outcome. Karyotyping all cases would lead to the diagnosis of all clinically significant abnormalities, and a policy of relying entirely on qf-PCR would lead to the diagnosis of 97.9% of abnormalities. An alternative strategy whereby qf PCR is the main method of analysis and full karyotyping is reserved for those cases with a minimum fetal NT thickness of 4 mm would require full karyotyping in 10.1% of the cases, would identify 99.0% of the significant abnormalities, and would cost 60% less than full karyotyping for all. CONCLUSIONS: In the diagnosis of chromosomal abnormalities after first trimester screening for trisomy 21, a policy of qf-PCR for all samples and karyotyping only if the fetal NT thickness is increased would reduce the economic costs, provide rapid delivery of results, and identify 99% of the clinically significant chromosomal abnormalities. PMID- 16476674 TI - Motor cortex plasticity in Parkinson's disease and levodopa-induced dyskinesias. AB - Experimental models of Parkinson's disease have demonstrated abnormal synaptic plasticity in the corticostriatal system, possibly related to the development of levodopa-induced dyskinesias (LID). We tested the hypothesis that LID in Parkinson's disease is associated with aberrant plasticity in the human motor cortex (M1). We employed the paired associative stimulation (PAS) protocol, an experimental intervention involving transcranial magnetic stimulation (TMS) and median nerve stimulation capable of producing long-term potentiation (LTP) like changes in the sensorimotor system in humans. We studied the more affected side of 16 moderately affected patients with Parkinson's disease (9 dyskinetic, 7 non dyskinetic) and the dominant side of 9 age-matched healthy controls. Motor-evoked potential (MEP) amplitudes and cortical silent period (CSP) duration were measured at baseline before PAS and for up to 60 min (T0, T30 and T60) after PAS in abductor pollicis brevis (APB) and abductor digiti minimi (ADM) muscles. PAS significantly increased MEP size in controls (+74.8% of baseline at T30) but not in patients off medication (T30: +0.07% of baseline in the non-dyskinetic, +27% in the dyskinetic group). Levodopa restored the potentiation of MEP amplitudes by PAS in the non-dyskinetic group (T30: +64.9% of baseline MEP) but not in the dyskinetic group (T30: -9.2% of baseline). PAS prolonged CSP duration in controls. There was a trend towards prolongation of CSP in the non-dyskinetic group off medications but not in the dyskinetic group. Levodopa did not restore CSP prolongation by PAS in the dyskinetic group. Our findings suggest that LTP like plasticity is deficient in Parkinson's disease off medications and is restored by levodopa in non-dyskinetic but not in dyskinetic patients. Abnormal synaptic plasticity in the motor cortex may play a role in the development of LID. PMID- 16476675 TI - Age-related differences in levodopa dynamics in Parkinson's: implications for motor complications. AB - Treatment-related motor complications in Parkinson's disease have been previously linked to disease-induced pre-synaptic alterations: dopaminergic denervation and changes in dopamine (DA) release patterns. The occurrence of such complications is also known to be partly dependent on the age of disease onset, occurring more frequently in patients with disease onset at a younger age. Using positron emission tomography (PET) and 4-h-long 18F-fluorodopa (FD) scans we have investigated in vivo an age dependence of disease-induced changes in DA turnover as a possible contributing factor to the age-related differences in treatment related motor complications. We evaluated the relative changes in DA turnover (measured by its direct inverse, effective DA distribution volume--EDV) and DA synthesis and vesicular storage capacity (quantified by the plasma input uptake rate constant Ki) in Parkinson's disease patients as a function of age (n = 27, age range 38-79 years). After correcting for disease severity, a significant negative correlation was found between age and magnitude of disease-induced decrease in EDV and in Ki in the putamen (P < 0.001, P = 0.02, respectively). However, the difference between the disease-induced decrease in EDV and that in Ki also exhibited an age dependence (P < 0.001), indicating a relatively higher disease-induced increase in DA turnover (inverse of EDV) compared with the decrease in DA synthesis and storage rate in patients of younger age compared with older patients. This finding implies that DA turnover in younger-onset patients undergoes a relatively greater alteration and thus likely contributes to a greater imbalance between DA synthesis, storage and release, which could lead to larger swings in synaptic DA levels. It has indeed been suggested on theoretical grounds that such imbalance may contribute to the greater propensity for motor fluctuations. These results provide one possible explanation for the age-dependent occurrence of complications and support the existence of a pre synaptic contribution to the occurrence of motor complications. PMID- 16476676 TI - Experimental contributions to the study of the embryology of the vagina. AB - BACKGROUND: Acien's hypothesis, deduced from patients with malformations of the female genital tract, especially those with renal agenesis and ipsilateral blind hemivagina, affirms the embryology of the human vagina as deriving from the Wolffian ducts and the Mullerian tubercle and could explain the embryological origin of all the female genital malformations reported. In this study, we investigated the hypothesis in rats. METHODS: Twenty-five pregnant rats were used to analyse female embryos (64) from day 15 (stage indifferent) to day 20 postcoitum (vagina completely formed). We performed transverse and longitudinal sections of embryos, haematoxylin-eosin tinction and immunohistochemical staining using markers specific to Wolffian derivatives. We also analysed the presence of these markers in the vagina of four adult rats. RESULTS: The Muller ducts converge until they fuse into one tube, but caudally they diverge and finally they fuse with the 'urogenital sinus bulbs' that are actually the distal portion of the Wolffian ducts according to the immunohistochemical marking with GZ1 and GZ2. The Mullerian tubercle is observed between those elements. Then, the immunohistochemical staining can be seen all along the completely formed vagina, which is also observed in the vagina of the adult rat. CONCLUSION: We prove the participation of Muller tubercle and Wolffian ducts in the formation of the vagina in rats, so we confirm experimentally Acien's hypothesis about the human vagina embryology. PMID- 16476677 TI - Detection of human immunodeficiency virus-1 RNA and DNA by extractive and in situ PCR in unprocessed semen and seminal fractions isolated by semen-washing procedure. AB - BACKGROUND: To determine the presence of human immunodeficiency virus-1 (HIV-1) viral RNA/DNA in whole semen, in properly isolated seminal fractions and in spermatozoa after swim-up, by extractive nested PCR and to compare the detection of HIV DNA by in situ PCR (IS-PCR) with the results of nested PCR. METHODS: We tested HIV-1 RNA and DNA by nested PCR in semen and in seminal fractions from 55 patients. Non-spermatic cells and spermatozoa pellet fractions from 10 HIV-1 positive and five HIV-1-negative men were tested for proviral DNA by IS-PCR. RESULTS: All samples of spermatozoa recovered after sperm washing were free of HIV RNA. HIV RNA tested positive in seven (13%) seminal plasma samples and only in two (4.2%) whole semen of these same samples. Of the seven seminal plasma samples testing positive for HIV RNA, four men had elevated blood viral load and three an undetectable viraemia. HIV DNA by IS-PCR turned positive in three of five samples in semen of HIV-noninfected men. CONCLUSION: HIV RNA/DNA detection in the semen of HIV-infected men proves the efficacy of sperm washing with swim up of spermatozoa. It is recommended that nested PCR be conducted on purified seminal compartments. IS-PCR is inadequate for detecting HIV in semen. PMID- 16476678 TI - Effects of transdermal testosterone application on the ovarian response to FSH in poor responders undergoing assisted reproduction technique--a prospective, randomized, double-blind study. AB - BACKGROUND: In primates, androgens can play a synergistic role with FSH in promoting the early follicular recruitment, which is critical in assisted reproduction technique programmes. OBJECTIVE: To assess whether poor responders can benefit from androgen application. METHODS: Inclusion criteria were a previous poor ovarian response to controlled ovarian stimulation and a decreased hormonal ovarian reserve. Selected women were randomized to receive either transdermal application of testosterone (n = 24) or placebo (n = 25) gel for 15 days before FSH treatment for a second IVF cycle. Similar GnRH analogue and equivalent FSH daily doses were used in both cycles. The primary outcome was the total number of oocytes retrieved. RESULTS: Testosterone gel application resulted in a significant increase in plasma testosterone levels but did not significantly improve the antral follicle count. Furthermore, after gel application, the main parameters of the ovarian response (numbers of pre-ovulatory follicles, total and mature oocytes and embryos) did not significantly differ between testosterone and placebo-treated patients. CONCLUSION: No significant beneficial effects of androgen administration on the ovarian response to FSH could be demonstrated. However, subsequent clinical trials are needed to determine whether an optimal dose and/or a longer duration of testosterone administration may be helpful. PMID- 16476679 TI - Routine use of hysterosalpingography prior to laparoscopy in the fertility workup: a multicentre randomized controlled trial. AB - BACKGROUND: A multicentre randomized controlled trial with or without hysterosalpingography (HSG) was conducted to assess the usefulness of HSG as a routine investigation in the fertility workup prior to laparoscopy and dye. METHODS: From 1 April 1997 to 1 April 2002, subfertile women were allocated by a computer-based 1 : 1 ratio randomization procedure, either for an HSG followed by laparoscopy and dye (the intervention group) of for laparoscopy and dye only (the control group) as a part of their fertility workup. Cumulative pregnancy rate (CPR) within 18 months after randomization was the primary outcome of interest. RESULTS: 344 women were randomized to the intervention group (n = 169) and the control group (n = 175). There was no significant difference in CPR at 18 months in the intervention group (49.1%) [95% confidence interval (CI) 41.6 to 56.6] and the control group (50.3%) (95% CI 42.8 to 57.8), a difference of -1.2% (95% CI 11.8% to 9.5%). CONCLUSION: The routine use of HSG at an early stage in the fertility workup prior to laparoscopy and dye does not influence CPR, compared with the routine use of laparoscopy and dye without HSG. PMID- 16476680 TI - Infant feeding with soy formula milk: effects on puberty progression, reproductive function and testicular cell numbers in marmoset monkeys in adulthood. AB - BACKGROUND: This marmoset study addresses concerns about feeding human male infants with soy formula milk (SFM). METHODS: From age 4 to 5 days, seven male co twin sets were fed standard formula milk (SMA) or SFM for 5-6 weeks; blood samples were subsequently collected at 10-week intervals. Testes from co-twins killed at 120-138 weeks were fixed for cell counts. RESULTS: SFM- and SMA-fed twins showed normal weight gain; puberty started and progressed normally, based on blood testosterone measurements. Body weight, organ weights (prostate, seminal vesicles, pituitary, thymus and spleen) and penis length were comparable in co twins. All SMA- and 6/7 SFM-fed males were fertile. Unexpectedly, testis weight (P = 0.041), Sertoli (P = 0.025) and Leydig cell (P = 0.026) numbers per testis were consistently increased in SFM-fed co-twins; the increase in Leydig cell numbers was most marked in males with consistently low-normal testosterone levels. Seminiferous epithelium volume per tubule showed a less consistent, non significant increase in SFM-fed males; raised germ cell numbers per testis, probably due to increased Sertoli cells, conceivably resulted in larger testes. Average lumen size, although greater in SFM-fed group, was inconsistent between co-twins and the difference was not significant. CONCLUSIONS: Infant feeding with SFM has no gross adverse reproductive effects in male marmosets, though it alters testis size and cell composition, and there is consistent, if indirect, evidence for possible 'compensated Leydig cell failure'. Similar and perhaps larger changes likely occur in adult men who were fed SFM as infants. PMID- 16476681 TI - Associations between relative income and mortality in Norway: a register-based study. AB - BACKGROUND: Current research on health inequalities suggests that not only an individual's absolute level of income but also his/her relative position in the income hierarchy could have health consequences. This study examines whether relative income was associated with individuals' mortality in Norway during the 1990s. METHODS: Data were formed by linkages of Norwegian administrative registers. This study analyses 1.68 million men and women (age group: 30-66 years) with disposable income (1993) in the range 60,000-210,000 Norwegian Kroner. Relative income was calculated as deviations in per cent from the median income in the surrounding residential area. The outcome variable was deaths in 1994-1999. Effects of relative income on mortality were estimated by multiple logistic regression analyses, separately in 13 narrow brackets of absolute income. Adjustments were made for sex, education, marital status, and other individual-level mortality predictors. RESULTS: Low relative income compared with the median in residential regions with populations above 20,000 inhabitants was associated with higher mortality among those with medium and lower absolute income. The excess risk increased progressively the lower the level of absolute income. Among those with higher absolute income, however, relative income was not associated with mortality. Moreover, when relative income was considered in relation to the median in small municipalities, almost no effect on mortality was observed. CONCLUSION: In Norway during the 1990s, having low relative income constituted an additional mortality risk among individuals with middle or lower absolute incomes and when relative income was calculated in relation to the average in medium-sized or larger regions. PMID- 16476682 TI - Alcohol and intimate partner violence: do we have enough information to act? AB - BACKGROUND: Male alcohol consumption is one of the accepted risk factors for intimate partner violence. The aim of this study is to assess the magnitude of the association between male alcohol consumption and intimate partner violence against women and the quality of the evidence of published papers exploring this relationship empirically. METHODS: Systematic review and meta-analysis of quantitative studies (1966-2004). Eight databases from Social and Behavioural Sciences, Clinical Medicine, and Life Sciences were reviewed. Studies with available 2 x 2 table or odds ratio were analysed using meta-analytic techniques. RESULTS: A total of 22 studies fulfilled the inclusion criteria for the systematic review: 14 (63.6%) were cross-sectional studies, 6 (27.3%) case series, 2 (9.1%) case-control studies. Ten studies analysed the relationship between alcohol and violence as their primary hypothesis and only two used a direct measure of alcohol consumption. Of them, 11 papers were included in the meta-analysis. The overall pooled odds ratio was 4.57 (95% confidence limits 3.30 6.35), but a high degree of heterogeneity was observed. The magnitude of the effect was inversely associated with the year of publication. The biggest odds ratios were obtained in the studies with the smallest sample sizes. CONCLUSIONS: The evidence about the relationship between alcohol consumption and intimate partner violence is of low quality in the study designs and maybe biased by publication of positive results. Currently there is not enough empirical evidence to support preventive policies based on male alcohol consumption as a risk factor in the particular case of intimate partner violence. PMID- 16476683 TI - Incidence and costs of injuries in The Netherlands. AB - BACKGROUND: Injuries are a major and persistent public health problem, but a comprehensive and detailed overview of the economic burden is missing. We therefore estimated the number of emergency department (ED) attendances and health care costs as a result of injury. METHODS: We estimated lifetime health care costs of injuries occurring in The Netherlands in the year 1999. Patient groups were defined that are homogeneous in terms of health service use. Health service use and costs per patient group was estimated with data from national databases and a prospective study among 5755 injury patients. RESULTS: Total health care costs due to injury in 1999 were euro 1.15 billion, or 3.7% of the total health care budget. Major cost peaks were observed among males between ages 15 and 44 due to a high incidence, and among females from age 65 onwards due to a high incidence and high costs per patient. For the age groups 0-14, 15-44, 45-64, and 65+ ED attendances per 1000 person years were 85, 85, 43, and 49, respectively, and costs per capita were euro 38, euro 59, euro 43, and euro 210, respectively. Costs per patient rise about linearly up to age 60 and about exponentially thereafter. From age 25 onwards, females have higher costs per patient than males. Hip fracture (20%), superficial injury (13%), open wounds (7%), and skull-brain injury (6%) had the highest total costs. Most costs were attributable to falls (44%) and traffic injuries (19%). CONCLUSION: Young adult males, elderly females, falls, hip fractures, and minor injuries without medical need for hospitalization account for a substantial share of health care costs. PMID- 16476684 TI - Prevention of mother-to-child transmission of HIV infection: Ukraine experience to date. AB - BACKGROUND: Despite the availability of effective interventions for the prevention of mother-to-child transmission (PMTCT), questions remain regarding implementation of programmes in settings with limited resources. This article sets out to describe the first 2 years of the implementation of the national PMTCT programme in Ukraine. METHODS: National data sources and data from a cohort of pregnant HIV-infected women delivering in 13 centres in Ukraine since 2000 were analysed. RESULTS: Interventions for prevention of MTCT have been implemented as a national programme within Ukraine's well developed infrastructure for maternal and child health. Implementation of an 'opt-out' model of counselling and HIV testing in antenatal clinics resulted in a 97% uptake of women who agreed to be HIV tested. In 2002, approximately 91% of HIV positive pregnant women received ARV prophylaxis (mainly single-dose nevirapine or short-course zidovudine) for PMTCT. The MTCT rate has decreased from 30% in 2000 to 10% in 2002. The need to scale-up prevention interventions in pregnant women with risky behaviour and late access to medical services was identified in a review of the national programme in 2003. CONCLUSIONS: Further implementation of a comprehensive approach for the prevention of HIV infection in infants, including more extensive ART regimen, as recommended by WHO, would help Ukraine to achieve the strategic goal of virtual elimination of HIV infection in infants by 2010. PMID- 16476685 TI - Lateral parabrachial lesions disrupt paraoxon-induced conditioned flavor avoidance. AB - Preliminary clinical evidence obtained in Gulf War veterans and patients suffering multiple chemical sensitivity points to the existence of a potential link between environmental exposure to organosphosphates (OPs) and the emergence of unspecific sickness syndromes in which associative Pavlovian conditioning might be partly involved. A laboratory animal model might be a useful tool for analyzing the involvement of conditioning in sickness syndromes potentially linked to OP poisoning. The first objective in the present study was to determine if paraoxon (PX), the neuroactive metabolite of the OP parathion, elicits a conditioned avoidance response to a novel stimulus (a taste-odor compound) in a conditioned flavor aversion procedure. Data obtained in Experiment 1 show conditioned flavor avoidance, demonstrative of the associative nature of the sickness properties of PX. The second objective was to characterize the nature of the specific physiological cue serving as the unconditioned stimulus in PX induced conditioned avoidance. Despite PX administration did induce cholinergic hyperactivity, as measured by body hypothermia and increased jaw movements, lesions of the lateral parabrachial area (lPB) disrupted PX-elicited flavor avoidance responses, indicating that cholinergic signs were not sufficient as unconditioned stimuli supporting avoidance responses. Given that lPB neural integrity is necessary to process aversive interoceptive information, disruption of conditioned flavor avoidance as a result of lPB lesions is consistent with a central interruption of interoceptive processing in PX-poisoned animals. Data are discussed under the light of the hypothesis claiming the importance of associative processes and noncholinesterase targets in sickness syndromes potentially induced by OP exposure. PMID- 16476686 TI - Research strategies for safety evaluation of nanomaterials, part VII: evaluating consumer exposure to nanoscale materials. AB - Considerable media attention has recently been given to novel applications for products that contain nanoscale materials. These products could have utility in several industries that market consumer products, including textiles, sporting equipment, cosmetics, consumer electronics, and household cleaners. Some of the purported benefits of these products include improved performance, convenience, lower cost, as well as other desirable features, when compared to the conventional products that do not contain nanoscale materials. Although there are numerous likely consumer advantages from products containing nanoscale materials, there is very little information available regarding consumer exposure to the nanoscale materials in these products or any associated risks from these exposures. This paper seeks to review a limited subset of products that contain nanoscale materials, assess the available data for evaluating the consumer exposures and potential hazards associated with these products, and discuss the capacity of U.S. regulatory agencies to address the potential risks associated with these products. PMID- 16476687 TI - A dose-response study following in utero and lactational exposure to di(2 ethylhexyl)phthalate: effects on female rat reproductive development. AB - Phthalates, a class of chemicals used as plasticizers, are economically important due to several industrial applications. Di(2-ethylhexyl)phthalate (DEHP) is the most commonly used phthalate plasticizer, and it has been described as a potent antiandrogen in males. We performed an extensive dose-response study following developmental exposure to DEHP and evaluated the effects on female reproductive development. Two wide ranges of doses that included dose levels relevant for human exposure as well as high doses typically used in toxicological studies were tested. Female Wistar rats were treated daily with DEHP and peanut oil (vehicle control) by gavage from gestation day 6 to lactation day 22. The low doses were 0.015, 0.045, 0.135, 0.405, and 1.215 mg DEHP/kg body weight (bw)/day, and the high doses were 5, 15, 45, 135, and 405 mg DEHP/kg bw/day. At the dose levels tested, no signs of maternal toxicity were observed. A significant delay in the age at vaginal opening (approximately 2 days) at 15 mg DEHP/kg bw/day and above, as well as a trend for a delay in the age at first estrus at 135 and 405 mg DEHP/kg bw/day (approximately 2 days), was observed. Liver enlargement (characteristic of phthalate exposure in rats) was limited to the 135- and 405-mg DEHP/kg bw/day doses. Anogenital distance and nipple development were unaffected. Based on the results of delayed pubertal onset, the no observed adverse effect level for female reproductive development may be set at 5 mg DEHP/kg bw/day. PMID- 16476688 TI - Distinct cytotoxic mechanisms of pristine versus hydroxylated fullerene. AB - The mechanisms underlying the cytotoxic action of pure fullerene suspension (nano C60) and water-soluble polyhydroxylated fullerene [C60(OH)n] were investigated. Crystal violet assay for cell viability demonstrated that nano-C60 was at least three orders of magnitude more toxic than C60(OH)n to mouse L929 fibrosarcoma, rat C6 glioma, and U251 human glioma cell lines. Flow cytometry analysis of cells stained with propidium iodide (PI), PI/annexin V-fluorescein isothiocyanate, or the redox-sensitive dye dihydrorhodamine revealed that nano-C60 caused rapid (observable after few hours), reactive oxygen species (ROS)-associated necrosis characterized by cell membrane damage without DNA fragmentation. In contrast, C60(OH)n caused delayed, ROS-independent cell death with characteristics of apoptosis, including DNA fragmentation and loss of cell membrane asymmetry in the absence of increased permeability. Accordingly, the antioxidant N-acetylcysteine protected the cell lines from nano-C60 toxicity, but not C60(OH)n toxicity, while the pan-caspase inhibitor z-VAD-fmk blocked C60(OH)n-induced apoptosis, but not nano-C60-mediated necrosis. Finally, C60(OH)n antagonized, while nano-C60 synergized with, the cytotoxic action of oxidative stress-inducing agents hydrogen peroxide and peroxynitrite donor 3-morpholinosydnonimine. Therefore, unlike polyhydroxylated C60 that exerts mainly antioxidant/cytoprotective and only mild ROS-independent pro-apoptotic activity, pure crystalline C60 seems to be endowed with strong pro-oxidant capacity responsible for the rapid necrotic cell death. PMID- 16476689 TI - A land plant-specific multigene family in the unicellular Mesostigma argues for its close relationship to Streptophyta. AB - The search for the unicellular relative of Streptophyta (i.e., land plants and their closest green algal relatives, the charophytes) started many years ago and remained centered around the scaly green flagellate, Mesostigma viride. To date, despite numerous studies, the phylogenetic position of Mesostigma is still debated and the nature of the unicellular ancestor of Streptophyta remains unknown. As molecular phylogenetic studies have produced conflicting results, we constructed a M. viride expressed sequence tags library and searched for sequences that are shared between M. viride and the Streptophyta (to the exclusion of the other green algal lineages--the Chlorophyta). Here, we report a multigene family that is restricted to Streptophyta and M. viride. The phylogenetic distribution of this complex character and its potential involvement in the evolution of an important land plant adaptive trait (i.e., three dimensional tissues) argue that Mesostigma is a close unicellular relative of Streptophyta. PMID- 16476690 TI - Strong asymmetric mutation bias in endosymbiont genomes coincide with loss of genes for replication restart pathways. AB - A large majority of bacterial genomes show strand asymmetry, such that G and T preferentially accumulate on the leading strand. The mechanisms are unknown, but cytosine deaminations are thought to play an important role. Here, we have examined DNA strand asymmetry in three strains of the aphid endosymbiont Buchnera aphidicola. These are phylogenetically related, have similar genomic GC contents, and conserved gene order structures, yet B. aphidicola (Bp) shows a fourfold higher replication-induced strand bias than B. aphidicola (Sg) and (Ap). We rule out an increase in the overall substitution frequency as the major cause of the stronger strand bias in B. aphidicola (Bp). Instead, the results suggest that the higher GC skew in this species is caused by a different spectrum of mutations, including a relatively higher frequency of C to T mutations on the leading strand and/or of G to A mutations on the lagging strand. A comparative analysis of 20 gamma-proteobacterial genomes revealed that endosymbiont genomes lacking recA and other genes involved in replication restart processes, such as priA, which codes for primosomal helicase PriA, displayed the strongest strand bias. We hypothesize that cytosine deaminations accumulate during single-strand exposure at arrested replication forks and that inefficient restart mechanisms may lead to high DNA strand asymmetry in bacterial genomes. PMID- 16476691 TI - Improving patient access to novel medical technologies in Europe. AB - The European Society of Cardiology (ESC) organized a one-day workshop with clinicians, health economic experts, and health technology appraisal experts to discuss the equity of patient access to novel medical technologies in Europe. Two index technologies were considered: implantable cardioverter defibrillators (ICDs) and drug-eluting stents (DES). The use of ICDs range from 35 implants/million population in Portugal to 166 implants/million population in Germany, whereas for implants of DES (as percentage of total stents) it is lowest in Germany at 14% and high in Portugal at 65%. These differences can in part be explained by a lack of structured implementation of guidelines, the direct cost in relation to the overall healthcare budget, and to differences in procedures and models applied by Health Technology Assessment (HTA) agencies in Europe. The workshop participants concluded that physicians need to be involved in a more structured way in HTA and need to become better acquainted with its methods and terminology. Clinical guidelines should be systematically translated, explained, disseminated, updated, and adopted by cardiologists in Europe. Clinically appropriate, consistent and transparent health economic models need to be developed and high-quality international outcome and cost data should be used. A process for funding of a technology should be developed after a positive recommendation from HTA agencies. Both the ESC and the national cardiac societies should build-up health economic expertise and engage more actively in discussions with stakeholders involved in the provision of healthcare. PMID- 16476692 TI - Diuretic usage in heart failure: a continuing conundrum in 2005. PMID- 16476693 TI - Markers of myocardial ischaemia. PMID- 16476695 TI - Neointimal proliferation around malapposed struts of a sirolimus-eluting stent: optical coherence tomography findings. PMID- 16476694 TI - Pharmacodynamics, pharmacokinetics, and safety of the oral reversible P2Y12 antagonist AZD6140 with aspirin in patients with atherosclerosis: a double-blind comparison to clopidogrel with aspirin. AB - AIMS: This double-blind, parallel-group study was conducted to assess the pharmacodynamics, pharmacokinetics, and safety of AZD6140, the first oral, reversible adenosine diphosphate (ADP) receptor antagonist. METHODS AND RESULTS: Patients (n = 200) with atherosclerosis were randomized to receive AZD6140 50, 100, or 200 mg twice daily (bid) or 400 mg daily (qd) or clopidogrel 75 mg qd for 28 days. All groups received aspirin 75-100 mg qd. AZD6140 (100 and 200 mg bid, 400 mg qd) rapidly and nearly completely inhibited ADP-induced platelet aggregation after initial dosing (day 1) and at day 28. On day 1, peak final extent inhibition of platelet aggregation (IPA) was observed 2-4 h post-dose with AZD6140, whereas clopidogrel minimally inhibited platelet aggregation (mean percentage IPA < 20%, all time points). Four hour post-dose at steady state, the three higher doses of AZD6140 produced comparable final-extent mean percentage IPA (approximately 90-95%), which exceeded that with AZD6140 50 mg bid or clopidogrel (approximately 60%). AZD6140 was generally well tolerated. All bleeding events, except one in a patient receiving 400 mg qd, were minor and of mild-to-moderate severity. CONCLUSION: AZD6140 100 and 200 mg bid were well tolerated and were superior to AZD6140 50 mg bid and clopidogrel 75 mg qd with regard to antiplatelet efficacy. PMID- 16476696 TI - Subcutaneous effusion resulting from an epidural catheter fragment. AB - The breakage of an epidural catheter within a patient is uncommon, but troublesome, complication of epidural block, and its cause is rarely discovered. In this case report, our aim was to present an effusion between s.c. tissue and fascia in the lumbar region because of a broken fragment of epidural catheter which was unnoticed during its removal. PMID- 16476697 TI - Patient-controlled cervical epidural fentanyl compared with patient-controlled i.v. fentanyl for pain after pharyngolaryngeal surgery. AB - BACKGROUND: Analgesia after pharyngolaryngeal surgery is commonly provided through the i.v. route. The aim of the study was to compare cervical epidural administration of fentanyl with the i.v. route for postoperative analgesia after pharyngolaryngeal surgery. METHODS: In a randomized double-blind study 42 patients received fentanyl via patient-controlled analgesia (PCA) either through the i.v. route (PCA-IV group, n=22) or through the cervical epidural route (PCA Epid group, n=20). Identical PCA settings were used in the two groups (bolus dose: 1.5 microg kg(-1), bolus: 25 microg, lockout interval: 10 min, maximum cumulative dose: 400 microg per 4 h). Analgesia at rest and during swallowing was evaluated using a visual analogue scale. RESULTS: Analgesia at rest was better in the PCA-Epid group than in the PCA-IV group but only 2 and 6 h after surgery (P<0.02). There was no difference in analgesia during swallowing. Cumulative doses of fentanyl were similar {PCA-Epid group: 1412 microg (912), PCA-IV group: 1287 microg (1200) [median (IQR)]}. The Pa(o(2)) showed a significant decrease between the preoperative and postoperative period, but this decrease was identical in the two groups [PCA-IV-group: 11.47 (2.4) kPa vs 8.27 (0.9) kPa; PCA Epid group: 11.33 (1.9) kPa vs 9.20 (2.4) kPa for preoperative and postoperative period respectively]. CONCLUSIONS: The study results show that cervical epidural analgesia provides marginally better pain relief at rest with no decrease in the fentanyl consumption. The use of the cervical epidural administration of fentanyl is questionable because of the possible complications of the technique. PMID- 16476698 TI - A comparison of the analgesic efficacy and side-effects of paravertebral vs epidural blockade for thoracotomy--a systematic review and meta-analysis of randomized trials. AB - Epidural analgesia is considered by many to be the best method of pain relief after major surgery. It is used routinely in many thoracic surgery centres. Although effective, side-effects include hypotension, urinary retention, incomplete (or failed) block, and, in rare cases, paraplegia. Paravertebral block (PVB) is an alternative technique that may offer comparable analgesic effectiveness and a better side-effect profile. We undertook a systematic review and meta-analysis of all relevant randomized trials comparing PVB with epidural analgesia in thoracic surgery. Data were abstracted and verified by both authors. Studies were tested for heterogeneity, and meta-analyses were done with random effects or fixed effects models. Weighted mean difference (WMD) was used for numerical outcomes and odds ratio (OR) for dichotomous outcomes, both with 95% CI. We identified 10 trials that had enrolled 520 thoracic surgery patients. All of the trials were small (n<130) and none were blinded. There was no significant difference between PVB and epidural groups for pain scores at 4-8, 24 or 48 h, WMD 0.37 (95% CI: -0.5, 121), 0.05 (-0.6, 0.7), -0.04 (-0.4, 0.3), respectively. Pulmonary complications occurred less often with PVB, OR 0.36 (0.14, 0.92). Urinary retention, OR 0.23 (0.10, 0.51), nausea and vomiting, OR 0.47 (0.24, 0.53), and hypotension, OR 0.23 (0.11, 0.48), were less common with PVB. Rates of failed block were lower in the PVB group, OR 0.28 (0.2, 0.6). PVB and epidural analgesia provide comparable pain relief after thoracic surgery, but PVB has a better side-effect profile and is associated with a reduction in pulmonary complications. PVB can be recommended for major thoracic surgery. PMID- 16476699 TI - 'Justifiable depression': how primary care professionals and patients view late life depression? A qualitative study. AB - BACKGROUND: Depression is the commonest mental health problem in elderly people and continues to be underdiagnosed and undertreated. AIM: To explore the ways that primary care professionals and patients view the causes and management of late-life depression. DESIGN: A qualitative study using semistructured interviews. SETTING: One Primary Care Trust in North West England. PARTICIPANTS: Fifteen primary care practitioners comprising nine GPs, three practice nurses, two district nurses and one community nurse; twenty patients who were over the age of 60 and who were participating in a feasibility study of a new model of care for late-life depression [PRIDE Trial: PRimary care Intervention for Depression in the Elderly (a feasibility study in Central Manchester funded by the Department of Health)]. RESULTS: Primary care practitioners conceptualized late-life depression as a problem of their everyday work, rather than as an objective diagnostic category. They described depression as part of a spectrum including loneliness, lack of social network, reduction in function and viewed depression as 'understandable' and 'justifiable'. This view was shared by patients. Therapeutic nihilism, the feeling that nothing could be done for this group of patients, was a feature of all primary care professionals' interviews. Patients' views were characterized by passivity and limited expectations of treatment. Depression was not viewed as a legitimate illness to be taken to the GP. Primary care professionals recognized that managing late-life depression did fall within their remit, but identified limitations in their own skills and capabilities in this area, as well as a lack of other resources to which they could refer patients. CONCLUSION: This study highlights the complicated nature of the diagnosis and management of late-life depression. Protocols for the diagnosis and treatment of depression emphasis the biomedical model which does not fit with the everyday experience of GPs or elderly patients who share the views of primary care professionals that depression is a consequence of social and contextual issues. There is a need for the development of evidence-based provision for older people with depression within primary care, but also a need for elderly patients to be made aware of the legitimacy of presenting low mood and misery to their primary care professional. PMID- 16476700 TI - Potential barriers to the use of health services among ethnic minorities: a review. AB - BACKGROUND: Ethnic minority patients seem to be confronted with barriers when using health services. Yet, care providers are often oblivious to these barriers, although they may share to some extent the burden of responsibility for them. In order to enlighten care providers, as to the potential pitfalls that may exist, there is a need to explore the different factors in the creation of the barriers. OBJECTIVE: Therefore, the objective of this paper is to present an overview of the potential barriers and the factors, which may restrict ethnic minority patients from using health services, according to the literature available. METHODS: Articles published from 1990 to 2003 were identified by searching electronic databases and selected through titles and abstracts. The articles were included if deemed to be relevant to study health services use by ethnic minorities, i.e. the different factors in the creation of a barrier. RESULTS: There were 54 articles reviewed. They reported on studies carried out in different countries and among different ethnic minorities. Potential barriers occurred at three different levels: patient level, provider level and system level. The barriers at patient level were related to the patient characteristics: demographic variables, social structure variables, health beliefs and attitudes, personal enabling resources, community enabling resources, perceived illness and personal health practices. The barriers at provider level were related to the provider characteristics: skills and attitudes. The barriers at system level were related to the system characteristics: the organisation of the health care system. CONCLUSION: This review has the goal of raising awareness about the myriad of potential barriers, so that the problem of barriers to health care for different ethnic minorities becomes transparent. In conclusion, there are many different potential barriers of which some are tied to ethnic minorities. The barriers are all tied to the particular situation of the individual patient and subject to constant adjustment. In other words, generalizations should not be made. PMID- 16476701 TI - The interaction of natural hepatitis C virus with human scavenger receptor SR BI/Cla1 is mediated by ApoB-containing lipoproteins. AB - The possible role of candidate receptors in the cellular penetration of HCV from serum of infected patients remains unclear. SR-BI/Cla1 interacts with plasma HDL, native and modified LDL, and VLDL, and facilitates cellular cholesterol efflux to lipoprotein acceptors. SR-BI/Cla1 binds HCV E2 protein and interacts with HCV pseudotypes via the HVR1 of the E2 envelope glycoprotein. Our data reveal that functional SR-BI/Cla1 expressed on the surface of CHO cells mediates the binding and uptake of HCV from the sera of infected patients. Interaction between HCV and SR-BI/Cla1 is not sensitive to either anti-E2 or anti-HVR1 antibodies but is effectively inhibited by anti-betalipoprotein antibodies and competed out by apoB containing lipoproteins and notably by VLDL. We interpret our data to indicate that VLDL associated with or incorporated into HCV plays a critical role in the primary interaction of HCV with SR-BI/Cla1, whereas the HCV E2 protein does not. In addition, our findings in hepatoma cell lines suggest that the interaction of HCV with human hepatocytes is equally mediated, at least in a part, by VLDL, and as such may represent an alternative pathway for infection. The association of HCV with ApoB-containing lipoproteins may promote cellular uptake of this virus in the presence of neutralizing antibodies. PMID- 16476702 TI - Sphingosine kinase functionally links elevated transmural pressure and increased reactive oxygen species formation in resistance arteries. AB - Myogenic vasoconstriction, an intrinsic response to elevated transmural pressure (TMP), requires the activation of sphingosine kinase (Sk1) and the generation of reactive oxygen species (ROS). We hypothesized that pressure-induced Sk1 signaling and ROS generation are functionally linked. Using a model of cannulated resistance arteries isolated from the hamster gracilis muscle, we monitored vessel diameter and smooth muscle cell (SMC) Ca2+i (Fura-2) or ROS production (dichlorodihydrofluorescein). Elevation of TMP stimulated the translocation of a GFP-tagged Sk1 fusion protein from the cytosol to the plasma membrane, indicative of enzymatic activation. Concurrently, elevation of TMP initiated a rapid and transient production of ROS, which was enhanced by expression of wild-type Sk1 (hSk(wt)) and inhibited by its dominant-negative mutant (hSk(G82D)). Exogenous sphingosine-1-phosphate (S1P) also stimulated ROS generation is isolated vessels. Chemical (1 micromol/L DPI), peptide (gp91ds-tat/gp91ds), and genetic (N17Rac) inhibition strategies indicated that NADPH oxidase was the source of the pressure induced ROS. NADPH oxidase inhibition attenuated myogenic vasoconstriction and reduced the apparent Ca2+ sensitivity of the SMC contractile apparatus, without affecting Ca2+-independent, RhoA-mediated vasoconstriction in response to exogenous S1P. Our results indicate a mandatory role for Sk1/S1P in mediating pressure-induced, NADPH oxidase-derived ROS formation. In turn, ROS generation appears to increase Ca2+ sensitivity, necessary for full myogenic vasoconstriction. PMID- 16476705 TI - Differential sialylation modulates voltage-gated Na+ channel gating throughout the developing myocardium. AB - Voltage-gated sodium channel function from neonatal and adult rat cardiomyocytes was measured and compared. Channels from neonatal ventricles required an approximately 10 mV greater depolarization for voltage-dependent gating events than did channels from neonatal atria and adult atria and ventricles. We questioned whether such gating shifts were due to developmental and/or chamber dependent changes in channel-associated functional sialic acids. Thus, all gating characteristics for channels from neonatal atria and adult atria and ventricles shifted significantly to more depolarized potentials after removal of surface sialic acids. Desialylation of channels from neonatal ventricles did not affect channel gating. After removal of the complete surface N-glycosylation structures, gating of channels from neonatal atria and adult atria and ventricles shifted to depolarized potentials nearly identical to those measured for channels from neonatal ventricles. Gating of channels from neonatal ventricles were unaffected by such deglycosylation. Immunoblot gel shift analyses indicated that voltage gated sodium channel alpha subunits from neonatal atria and adult atria and ventricles are more heavily sialylated than alpha subunits from neonatal ventricles. The data are consistent with approximately 15 more sialic acid residues attached to each alpha subunit from neonatal atria and adult atria and ventricles. The data indicate that differential sialylation of myocyte voltage gated sodium channel alpha subunits is responsible for much of the developmental and chamber-specific remodeling of channel gating observed here. Further, cardiac excitability is likely impacted by these sialic acid-dependent gating effects, such as modulation of the rate of recovery from inactivation. A novel mechanism is described by which cardiac voltage-gated sodium channel gating and subsequently cardiac rhythms are modulated by changes in channel-associated sialic acids. PMID- 16476707 TI - Absence of alpha 7 integrin in dystrophin-deficient mice causes a myopathy similar to Duchenne muscular dystrophy. AB - Both the dystrophin-glycoprotein complex and alpha7beta1 integrin have critical roles in the maintenance of muscle integrity via the provision of mechanical links between muscle fibres and the basement membrane. Absence of either dystrophin or alpha7 integrin results in a muscular dystrophy. To clarify the role of alpha7 integrin and dystrophin in muscle development and function, we generated integrin alpha7/dystrophin double-mutant knockout (DKO) mice. Surprisingly, DKO mice survived post-natally and were indistinguishable from wild type, integrin alpha7-deficient and mdx mice at birth, but died within 24-28 days. Histological analysis revealed a severe muscular dystrophy in DKO mice with endomysial fibrosis and ectopic calcification. Weight loss was correlated with the loss of muscle fibres, indicating that progressive muscle wasting in the double mutant was most likely due to inadequate muscle regeneration. The data further support that premature death of DKO mice is due to cardiac and/or respiratory failure. The integrin alpha7/dystrophin-deficient mouse model, therefore, resembles the pathological changes seen in Duchenne muscular dystrophy and suggests that the different clinical severity of dystrophin deficiency in human and mouse may be due to a fine-tuned difference in expression of dystrophin and integrin alpha7 in both species. Together, these findings indicate an essential role for integrin alpha7 in the maintenance of dystrophin-deficient muscles. PMID- 16476708 TI - Incidence, time course, and determinants of menstrual bleeding after breast cancer treatment: a prospective study. AB - PURPOSE: To assess ovarian function using the surrogate of monthly bleeding after breast cancer treatment in premenopausal women. PATIENTS AND METHODS: Five hundred ninety-five US women age 20 to 45 years were accrued from January 1998 to July 2002 within 8 months of diagnosis with stages I to III breast cancer (median follow-up 45 months). Daily bleeding records were obtained prospectively, as well as extensive clinical, demographic, quality of life, and treatment data. Repeated measures logistic regression was used to assess which variables were predictive of monthly bleeding. RESULTS: Significantly different proportions of women had monthly bleeding depending on their age (P < .001), chemotherapy program (P < .001), and time since treatment regimen. In the month after the standard course of doxorubicin and cyclophosphamide (AC), whether or not followed by paclitaxel or docetaxel, approximately 16% had monthly bleeding compared with the cyclophosphamide, methotrexate, fluorouracil (CMF) group, in which 48% bled (P < .001). Following any AC regimen, there was a slow recovery phase of about 9 months followed by a plateau, during which almost half continued monthly bleeding for the remainder of the follow-up period compared with after CMF in which there was no recovery phase and a continual decline in monthly bleeding to approximately 18% of women at study end (P < .001). Tamoxifen use decreased bleeding between months 12 and 24 after chemotherapy with 15% fewer women having bleeding. CONCLUSION: Using daily menstrual bleeding records, it is demonstrated that age, the specific chemotherapy regimen received, and tamoxifen use impact ovarian function. PMID- 16476706 TI - Association between two mu-opioid receptor gene (OPRM1) haplotype blocks and drug or alcohol dependence. AB - We examined 13 single nucleotide polymorphisms (SNPs) spanning the coding region of the mu-opioid receptor gene (OPRM1), among 382 European Americans (EAs) affected with substance dependence [alcohol dependence (AD) and/or drug dependence (DD)] and 338 EA healthy controls. These SNPs delineated two haplotype blocks. Genotype distributions for all SNPs were in Hardy-Weinberg equilibrium (HWE) in controls, but in cases, four SNPs in Block I and three SNPs in Block II showed deviation from HWE. Significant differences were found between cases and controls in allele and/or genotype frequencies for six SNPs in Block I and two SNPs in Block II. Association of SNP4 in Block I with DD (allele: P=0.004), SNP5 in Block I with AD and DD (allele: P< or =0.005 for both) and two SNPs in Block II with AD (SNP11 genotype: P=0.002; SNP12 genotype: P=0.001) were significant after correction for multiple testing. Frequency distributions of haplotypes (constructed by five tag SNPs) differed significantly for cases and controls (P<0.001 for both AD and DD). Logistic regression analyses confirmed the association between OPRM1 variants and substance dependence, when sex and age of subjects and alleles, genotypes, haplotypes or diplotypes of five tag SNPs were considered. Population structure analyses excluded population stratification artifact. Additional supporting evidence for association between OPRM1 and AD was obtained in a smaller Russian sample (247 cases and 100 controls). These findings suggest that OPRM1 intronic variants play a role in susceptibility to AD and DD in populations of European ancestry. PMID- 16476710 TI - Possible role for tumour necrosis factor inhibitors in the treatment of resistant dermatomyositis and polymyositis: a retrospective study of eight patients. AB - OBJECTIVE: To understand the use of tumour necrosis factor (TNF)alpha inhibitors in refractory dermatomyositis and polymyositis in an academic centre. METHODS: A retrospective study of eight patients with dermatomyositis or polymyositis refractory to corticosteroids and immunosuppressives who were treated with TNF inhibitors between 1998 and 2004. RESULTS: 8 patients with dermatomyositis or polymyositis who were treated with TNF inhibitors as adjunct treatment were identified. The mean (SD) duration of disease before initiation of TNF inhibitors was 8.5 (4.4) years. The patients failed to respond to treatment with corticosteroids (oral and intravenous); intravenous immunoglobulin and immunosuppressants (methotrexate, azathioprine, mycophenolate mofetil and leflunomide); 4.5 (1.4) immunosuppressants had been used before TNF treatment. Six patients were treated with etanercept alone, one with infliximab and one sequentially with both agents. Of the eight patients, six showed a favourable response with improved motor strength and decreased fatigue after 15.2 (6.5) months. Two of the patients did not respond after 4 (1.4) months and TNF inhibitors were discontinued. Responders showed a 54.4% (27.7%) decrease in serum concentration of creatine kinase, which was grossly abnormal (4463.5 (4036.4) U/l). Non-responders had similar reductions in creatine kinase concentration (56.1% (20.4%)), but their pre-treatment concentrations were in the normal range (118.5 (19.1) U/l). CONCLUSION: Anti-TNF agents may be useful in some patients with refractory dermatomyositis or polymyositis. PMID- 16476711 TI - Epistatic interaction between FCRL3 and NFkappaB1 genes in Spanish patients with rheumatoid arthritis. AB - BACKGROUND: A Japanese study has described a strong association between rheumatoid arthritis and several polymorphisms located in the Fc receptor-like 3 (FCRL3) gene, a member of a family of genes related to Fc receptors located on chromosome 1q21-23. OBJECTIVES: To evaluate the association between rheumatoid arthritis and FCLR3 polymorphisms in a large cohort of Caucasian patients with rheumatoid arthritis and healthy controls of Spanish origin. Owing to the described functional link between the FCRL3 polymorphisms and the transcription factor nuclear factor kappaB (NFkappaB), a functional polymorphism located in the NFkappaB1 gene was included. METHODS: 734 patients with rheumatoid arthritis from Madrid and Granada, Spain, were included in the study, along with 736 healthy controls. Polymorphisms in the FCRL3 gene were studied by TaqMan technology. The 94ins/delATTG NFkappaB1 promoter polymorphism was analysed by fragment analysis after polymerase chain reaction with labelled primers. Genotypes were compared using 3x2 contingency tables and chi2 values. RESULTS: No overall differences were found in any of the FCRL3 polymorphisms and in the NFkappaB1 promoter polymorphism when patients were compared with controls. However, when stratified according to NFkappaB1 genotypes, a susceptibility effect of FCRL3 polymorphisms was observed in patients who were heterozygotes for NFkappaB1 (pc = 0.003). CONCLUSIONS: The FCRL3 polymorphisms associated with rheumatoid arthritis in a Japanese population are not associated per se with rheumatoid arthritis in a Spanish population. A genetic interaction was found between NFkappaB1 and FCRL3 in Spanish patients with rheumatoid arthritis. These findings may provide a general rationale for divergent genetic association results in different populations. PMID- 16476709 TI - Cutaneous abnormalities in rheumatoid arthritis compared with non-inflammatory rheumatic conditions. AB - BACKGROUND: Cutaneous abnormalities are common in rheumatoid arthritis, but exact prevalence estimates are yet to be established. Some abnormalities may be independent and coincidental, whereas others may relate to rheumatoid arthritis or its treatment. OBJECTIVES: To determine the exact nature and point prevalence of cutaneous abnormalities in patients with rheumatoid arthritis compared with those in patients with non-inflammatory rheumatic disease. METHODS: 349 consecutive outpatients for rheumatology (205 with rheumatoid arthritis and 144 with non-inflammatory rheumatic conditions) were examined for skin and nail signs by a dermatologist. Histories of rheumatology, dermatology, drugs and allergy were noted in detail. RESULTS: Skin abnormalities were reported by more patients with rheumatoid arthritis (61%) than non-inflammatory controls (47%). More patients with rheumatoid arthritis (39%) than controls (10%) attributed their skin abnormality to drugs. Cutaneous abnormalities observed by the dermatologist were also more common in patients with rheumatoid arthritis (76%) than in the group with non-inflammatory disease (60%). Specifically, bruising, athlete's foot, scars, rheumatoid nodules and vasculitic lesions were more common in patients with rheumatoid arthritis than in controls. The presence of bruising was predicted only by current steroid use. The presence of any other specific cutaneous abnormalities was not predicted by any of the variables assessed. In the whole group, current steroid use and having rheumatoid arthritis were the only important predictors of having any cutaneous abnormality. CONCLUSIONS: Self reported and observed cutaneous abnormalities are more common in patients with rheumatoid arthritis than in controls with non-inflammatory disease. These include cutaneous abnormalities related to side effects of drugs or to rheumatoid arthritis itself and other abnormalities previously believed to be independent but which may be of clinical importance. PMID- 16476712 TI - Altered gut transcriptome in spondyloarthropathy. AB - BACKGROUND: Intestinal inflammation is a common feature of spondyloarthropathy (SpA) and Crohn's disease. Inflammation is manifested clinically in Crohn's disease and subclinically in SpA. However, a fraction of patients with SpA develops overt Crohn's disease. AIMS: To investigate whether subclinical gut lesions in patients with SpA are associated with transcriptome changes comparable to those seen in Crohn's disease and to examine global gene expression in non inflamed colon biopsy specimens and screen patients for differentially expressed genes. METHODS: Macroarray analysis was used as an initial genomewide screen for selecting a comprehensive set of genes relevant to Crohn's disease and SpA. This led to the identification of 2625 expressed sequence tags that are differentially expressed in the colon of patients with Crohn's disease or SpA. These clones, with appropriate controls (6779 in total), were used to construct a glass-based microarray, which was then used to analyse colon biopsy specimens from 15 patients with SpA, 11 patients with Crohn's disease and 10 controls. RESULTS: 95 genes were identified as differentially expressed in patients with SpA having a history of subclinical chronic gut inflammation and also in patients with Crohn's disease. Principal component analysis of this filtered set of genes successfully distinguished colon biopsy specimens from the three groups studied. Patients with SpA having subclinical chronic gut inflammation cluster together and are more related to those with Crohn's disease. CONCLUSION: The transcriptome in the intestine of patients with SpA differs from that of controls. Moreover, these gene changes are comparable to those seen in patients with Crohn's disease, confirming initial clinical observations. On the basis of these findings, new (genetic) markers for detection of early Crohn's disease in patients with SpA can be considered. PMID- 16476714 TI - Fatal Candida famata peritonitis complicating sclerosing peritonitis in a peritoneal dialysis patient. PMID- 16476715 TI - Thrombosis of vascular access associated with factor V Leiden, antiphospholipid antibodies and antiheparin antibodies in a young woman on dialysis receiving warfarin following mitral valve replacement. PMID- 16476713 TI - Long term NSAID treatment inhibits COX-2 synthesis in the knee synovial membrane of patients with osteoarthritis: differential proinflammatory cytokine profile between celecoxib and aceclofenac. AB - OBJECTIVE: To compare the effect of celecoxib with that of a classic non steroidal anti-inflammatory drug (NSAID) on synovial inflammation and on the synovial expression of proinflammatory genes in patients with knee osteoarthritis (OA). METHODS: 30 patients with severe knee OA scheduled for total knee replacement surgery were included in a 3 month clinical trial. They were randomised to two groups: patients treated with celecoxib (CBX) (200 mg/24 h) and patients treated with aceclofenac (ACF) (100 mg/12 h). Those patients with OA who did not want to be treated with NSAIDs served as a control group. During knee surgery, synovial fluid (SF) and synovial membrane (SM) were collected. A SM specimen was fixed and embedded in paraffin and another part was frozen for molecular biology studies. RESULTS: At the end of study both CBX and ACF treated patients showed a significant improvement in pain and knee function compared with controls. Both drugs significantly reduced prostaglandin E(2) (PGE(2)) SF concentration and down regulated COX-2 mRNA and protein expression at the SM. However, synovial macrophage infiltration (CD68 antigen staining) and expression of proinflammatory mediators, such as interleukin 1beta and tumour necrosis factor alpha, were decreased only by CBX treatment. CONCLUSION: Both drugs improved joint pain and function, inhibited SF PGE(2) concentration, and induced a decrease in synovial COX-2 expression and synthesis not related to the tissue inflammatory status. These data suggest that PGE(2) blocking agents may decrease PGE(2) production not only by direct COX-2 inhibition but also by down regulating COX-2 expression and synthesis. However, CBX and ACF appear to have different anti-inflammatory profiles in controlling OA synovial macrophage infiltration and proinflammatory expression. PMID- 16476717 TI - Renoprotection of angiotensin receptor blockers: beyond blood pressure lowering. PMID- 16476716 TI - Hypergonadotropic hypogonadism and renal failure due to WT1 mutation. PMID- 16476718 TI - Increased expression of monocytic angiotensin-converting enzyme in dialysis patients with cardiovascular disease. AB - BACKGROUND: Patients with chronic renal disease suffer from accelerated atherogenesis, which is promoted by inflammation and oxidative stress. Tissue angiotensin converting enzyme (ACE) exerts proinflammatory and prooxidative effects by producing angiotensin II. Monocytes are strongly involved in the pathogenesis of atherosclerosis. They express ACE, which might contribute to their atherogenic potency. We hypothesize that dialysis patients have increased monocytic ACE expression, and that ACE expression on circulating monocytes is associated with prevalent cardiovascular disease. METHODS: In 74 dialysis patients, ACE expression on total monocytes and monocyte subsets was measured flow-cytometrically in a whole-blood assay. A subpopulation of 22 dialysis patients was compared to an age- and gender-matched control group with intact renal function. Cardiovascular risk factors and the prevalence of cardiovascular disease were assessed. In a subgroup of patients (n = 8), monocytic ACE activity was measured in vitro and correlated with monocytic ACE expression. RESULTS: Dialysis patients had an increased expression of monocytic ACE compared to controls. Monocytic ACE expression was higher in dialysis patients with prevalent cardiovascular disease than in those without cardiovascular disease. This association remained significant after correction for classical cardiovascular risk factors. Among monocyte subsets, CD14++CD16+ monocytes had the highest ACE expression. Monocytic ACE activity correlated with ACE surface expression. CONCLUSIONS: The finding of increased ACE expression on monocytes of dialysis patients with cardiovascular disease links monocytes to the activated renin angiotensin system. ACE expression was found highest among CD14++16+ monocytes, which is in accordance with a prominent role of these proinflammatory cells in atherogenesis. PMID- 16476719 TI - Respective role of uraemic toxins and myeloperoxidase in the uraemic state. AB - BACKGROUND: In haemodialysis (HD) patients, advanced oxidation protein products (AOPP) were previously ascribed to oxidized plasma proteins, resulting mainly from increased myeloperoxidase (MPO) activity. The aim of the present study was to assess the mechanisms leading to the generation of AOPP during the course of chronic kidney disease including end-stage renal disease, with particular focus on AOPP and MPO characterization in the plasma at decreasing levels of kidney function. METHODS: Phagocyte activation was evaluated by whole blood NADPH oxidase and MPO activities. In plasma, MPO protein concentration was quantified by ELISA and catalytic activity assayed by the spectrophotometric detection of phenol and 4-aminoantipyrine (AAP) co-oxidation in the presence of hydrogen peroxide (H(2)O(2)). RESULTS: In HD patients, plasma AOPP concentration was linked to neutrophil oxidative activity. Such an association was not found in control subjects or predialysis patients, suggesting that in the latter, AOPP generation did not mainly result from MPO released by activated neutrophils. Similarly, plasma AOPP correlated with plasma MPO protein concentration in HD patients, but not in control subjects or predialysis patients, suggesting that in the latter AOPP did not predominantly result from MPO activity. This interpretation was supported by the observation of a greater degree of co oxidation of phenol and AAP in the absence of H(2)O(2) in predialysis patients than in HD patients or control subjects. The contribution of MPO dramatically differed between predialysis and HD patients (2+/-5 vs 46+/-6%; P<0.001). CONCLUSION: Our observations suggest that AOPP generation in predialysis patients mainly results from MPO-independent oxidation mechanisms. PMID- 16476721 TI - Severity scoring and mortality 1 year after acute renal failure. PMID- 16476720 TI - Soluble adhesion molecules in end-stage renal disease: a predictor of outcome. AB - BACKGROUND: Inflammation is thought to contribute to initiation and aggravation of atherosclerosis through a process predominantly mediated by adhesion molecules. The aims of this study were to investigate the association between the concentrations of circulating soluble intercellular (sICAM-1) and vascular cellular (sVCAM-1) adhesion molecules and clinical outcome, and to evaluate the effect of antihypertensive drugs on sICAM-1 and sVCAM-1 concentrations in end stage renal disease (ESRD) patients. METHODS: We prospectively investigated 310 (191 males) incident ESRD patients, 53+/-12 years old, shortly before the start of renal replacement therapy. Glomerular filtration rate (GFR) was 6.4 (range 0.8 16.5) ml/min/1.73 m(2). Plasma sICAM-1 and sVCAM-1 were measured by enzyme-linked immunosorbent assay (ELISA) kits. Survival was determined from the day of examination, with a mean follow-up period of 39 (range 1-123) months. RESULTS: In non-adjusted analysis, high sICAM-1 and sVCAM-1 levels were associated with all cause and cardiovascular (P<0.001) mortality. After adjusting for age, gender, diabetes mellitus, serum cholesterol, C-reactive protein (CRP), subjective global assessment and angiotensin-converting enzyme inhibitors (ACEI) or angiotensin II receptor blockers (ARB), the association between high sICAM-1 and mortality remained significant for all-cause (HR 1.9; CI 1.2-2.9, P = 0.004) and cardiovascular (HR 1.8; CI 1.1-3.1, P = 0.02) mortality, and a high sVCAM-1 was associated with all-cause mortality (HR 1.7; CI 1.04-2.7, P = 0.03). Furthermore, the concentration of sICAM-1, but not sVCAM-1, was lower in patients receiving ACEI/ARB (254+/-83 vs 275+/-92 ng/ml; P<0.05) or patients receiving calcium channel blockers (CCB, 251+/-75 vs 273+/-95 ng/ml; P<0.05) than in non-users. CONCLUSIONS: In ESRD patients, sICAM-1 and sVCAM-1 are independent predictors of all cause and cardiovascular death. The use of ACEI/ARB or CCB was associated with decreased concentrations of soluble adhesion molecules. PMID- 16476722 TI - The role of pre-emptive re-transplant in graft and recipient outcome. AB - BACKGROUND: The effect of the pre-emptive re-transplant, and of inter-transplant waiting time generally, on graft and recipient survival is not well established. METHODS: Analysis of the United States Renal Data System (USRDS) data (1/1/90 through 12/31/00; n = 92,844) was performed. Cox regression was used to analyse time to event, with an additional analysis to stratify by transplant era. RESULTS: Having a prior transplant, as well as the total number of transplants, was related to an increased risk of graft failure [hazard ratio (HR) 1.24, P<0.001 for history of prior transplant; HR 1.35 per transplant, P<0.001], but not to recipient death. The time waiting for re-transplant slightly worsened the risk for recipient mortality in the entire patient population and in the recipients of single re-transplant (HR 1.003 and 1.004 per month respectively, P<0.001), and for graft failure only in recipients of single re-transplant (HR 1.001 per month, P<0.05). Pre-emptive re-transplant (dialysis-free re-transplant or transplant within 6 days of last graft failure) increased the risk of graft failure (HR 1.36, P<0.001) and did not have any statistically significant effect on recipient survival. The longer duration of prior graft survival but not the type of the graft (living vs deceased) had protective effect on the consecutive graft and recipient survival. CONCLUSIONS: With the potential caveats associated with retrospective data analysis, these results suggest that pre-emptive re transplantation is associated with increased risk of graft failure, while longer time on dialysis in between transplants is associated with negative effect upon graft and recipient survival in most patient subgroups. The optimal time in between graft failure and re-transplant was not evaluated in this study. Further prospective studies might be needed to confirm the observed effects. PMID- 16476723 TI - Delayed myeloid engraftment due to vancomycin in allogeneic haematopoietic stem cell transplant recipients. PMID- 16476724 TI - Inter-country transfer of Gram-negative organisms carrying the VIM-4 and OXA-58 carbapenem-hydrolysing enzymes. PMID- 16476725 TI - Heterologous expression of glycopeptide resistance vanHAX gene clusters from soil bacteria in Enterococcus faecalis. AB - OBJECTIVES: The aim of the study was to determine whether glycopeptide resistance gene clusters from soil bacteria could be heterologously expressed in Enterococcus faecalis and adapt to the new host following exposure to vancomycin. METHODS: The vanHAX clusters from Paenibacillus thiaminolyticus PT-2B1, Paenibacillus apiarius PA-B2B and Amycolatopsis coloradensis DSM 44225 were separately cloned in an appropriately constructed shuttle vector containing the two-component regulatory system (vanRS) of Tn1546. The complete vanA(PT) operon (vanRSHAXY) from P. thiaminolyticus PT-2B1 was cloned in the same shuttle vector lacking enterococcal vanRS. All plasmid constructs were electroporated into E. faecalis JH2-2 and the MICs of vancomycin and teicoplanin were determined for each recombinant strain before and following exposure to sublethal concentrations of vancomycin. RESULTS: The vanHAX clusters from P. thiaminolyticus and P. apiarius conferred high-level vancomycin resistance (MIC > or = 125 mg/L) in E. faecalis JH2-2. In contrast, cloning of the vanHAX cluster from A. coloradensis did not result in a significant increase of vancomycin resistance (MIC = 0.7 mg/L). Resistance to vancomycin was not observed after cloning the complete vanA(PT) operon from P. thiaminolyticus (MIC = 2 mg/L), but this recombinant rapidly adapted to high concentrations of vancomycin (MIC = 500 mg/L) following exposure to sub-lethal concentrations of this antibiotic. CONCLUSION: The results showed that vanA(PT) in P. thiaminolyticus is a possible ancestor of vanA mediated glycopeptide resistance in enterococci. Experimental evidence supported the hypothesis that enterococci did not acquire glycopeptide resistance directly from glycopeptide-producing organisms such as A. coloradensis. PMID- 16476726 TI - Prohormone convertase 1/3 is essential for processing of the glucose-dependent insulinotropic polypeptide precursor. AB - The physiology of the incretin hormones, glucagon-like peptide 1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP), and their role in type 2 diabetes currently attract great interest. Recently we reported an essential role for prohormone convertase (PC) 1/3 in the cleavage of intestinal proglucagon, resulting in formation of GLP-1, as demonstrated in PC1/3-deficient mice. However, little is known about the endoproteolytic processing of the GIP precursor. This study investigates the processing of proGIP in PC1/3 and PC2 null mice and in cell lines using adenovirus-mediated overexpression. Supporting a role for PC1/3 in proGIP processing, we found co-localization of GIP and PC1/3 but not PC2 in intestinal sections by immunohistochemistry, and analysis of intestinal extracts from PC1/3-deficient animals demonstrated severely impaired processing to GIP, whereas processing to GIP was unaltered in PC2-deficient mice. Accordingly, overexpression of preproGIP in the neuroendocrine AtT-20 cell line that expresses high levels of endogenous PC1/3 and negligible levels of PC2 resulted in production of GIP. Similar results were obtained after co-expression of preproGIP and PC1/3 in GH4 cells that express no PC2 and only low levels of PC1/3. In addition, studies in GH4 cells and the alpha-TC1.9 cell line, expressing PC2 but not PC1/3, indicate that PC2 can mediate processing to GIP but also to other fragments not found in intestinal extracts. Taken together, our data indicate that PC1/3 is essential and sufficient for the production of the intestinal incretin hormone GIP, whereas PC2, although capable of cleaving proGIP, does not participate in intestinal proGIP processing and is not found in intestinal GIP-expressing cells. PMID- 16476727 TI - Enhancement of +1 frameshift by polyamines during translation of polypeptide release factor 2 in Escherichia coli. AB - Polypeptide release factor 2 (RF2) in Escherichia coli is known to be synthesized by a +1 frameshift at the 26th UGA codon of RF2 mRNA. Polyamines were found to stimulate the +1 frameshift of RF2 synthesis, an effect that was reduced by excess RF2. Polyamine stimulation of +1 frameshift of RF2 synthesis was observed at the early logarithmic phase, which is the important phase in determination of the overall rate of cell growth. A Shine-Dalgarno-like sequence was necessary for an efficient +1 frameshift of RF2 synthesis, but not for polyamine stimulation. Spectinomycin, tetracycline, streptomycin, and neomycin reduced polyamine stimulation of the +1 frameshift of RF2 synthesis. The results suggest that a structural change of the A site on 30 S ribosomal subunits is important for polyamine stimulation of the +1 frameshift. The level of mRNAs of ribosomal proteins and elongation factors having UAA as termination codon was enhanced by polyamines, and OppA synthesis from OppA mRNA having UAA as termination codon was more enhanced by polyamines than that from OppA mRNA having a UGA termination codon. Furthermore, synthesis of ribosomal protein L20 and elongation factor G from the mRNAs having a UAA termination codon was enhanced by polyamines at the level of translation and transcription. The results suggest that some protein synthesis from mRNAs having a UAA termination codon is enhanced at the level of translation through polyamine stimulation of +1 frameshift of RF2 synthesis. It is concluded that prfB encoding RF2 is a new member of the polyamine modulon. PMID- 16476729 TI - Crystal structure and nonhomologous end-joining function of the ligase component of Mycobacterium DNA ligase D. AB - DNA ligase D (LigD) is a large polyfunctional enzyme involved in nonhomologous end-joining (NHEJ) in mycobacteria. LigD consists of a C-terminal ATP-dependent ligase domain fused to upstream polymerase and phosphoesterase modules. Here we report the 2.4 angstroms crystal structure of the ligase domain of Mycobacterium LigD, captured as the covalent ligase-AMP intermediate with a divalent metal in the active site. A chloride anion on the protein surface coordinated by the ribose 3'-OH and caged by arginine and lysine side chains is a putative mimetic of the 5'-phosphate at a DNA nick. Structure-guided mutational analysis revealed distinct requirements for the adenylylation and end-sealing reactions catalyzed by LigD. We found that a mutation of Mycobacterium LigD that ablates only ligase activity results in decreased fidelity of NHEJ in vivo and a strong bias of mutagenic events toward deletions instead of insertions at the sealed DNA ends. This phenotype contrasts with the increased fidelity of double-strand break repair in deltaligD cells or in a strain in which only the polymerase function of LigD is defective. We surmise that the signature error-prone quality of bacterial NHEJ in vivo arises from a dynamic balance between the end-remodeling and end sealing steps. PMID- 16476728 TI - Conformational stabilities of the structural repeats of erythroid spectrin and their functional implications. AB - The two polypeptide chains of the erythroid spectrin heterodimer contain between them 36 structural repeating modules, which can function as independently folding units. We have expressed all 36 and determined their thermal stabilities. These vary widely, with unfolding transition mid-points (T(m)) ranging from 21 to 72 degrees C. Eight of the isolated repeats are largely unfolded at physiological temperature. Constructs comprising two or more adjacent repeats show inter-repeat coupling with coupling free energies of several kcal mol(-1). Constructs comprising five successive repeats from the beta-chain displayed cooperativity and strong temperature dependence in forced unfolding by atomic force microscopy. Analysis of aligned sequences and molecular modeling suggests that high stability is conferred by large hydrophobic side chains at position e of the heptad hydrophobic repeats in the first helix of the three-helix bundle that makes up each repeat. This inference was borne out by the properties of mutants in which the critical residues have been replaced. The marginal stability of the tertiary structure at several points in the spectrin chains is moderated by energetic coupling with adjoining structural elements but may be expected to permit adaptation of the membrane to the large distortions that the red cell experiences in the circulation. PMID- 16476730 TI - Role of heme iron coordination and protein structure in the dynamics and geminate rebinding of nitric oxide to the H93G myoglobin mutant: implications for nitric oxide sensors. AB - The influence of the heme iron coordination on nitric oxide binding dynamics was investigated for the myoglobin mutant H93G (H93G-Mb) by picosecond absorption and resonance Raman time-resolved spectroscopies. In the H93G-Mb, the glycine replacing the proximal histidine does not interact with the heme iron so that exogenous substituents like imidazole may coordinate to the iron at the proximal position. Nitrosylation of H93G-Mb leads to either 6- or 5-coordinate species depending on the imidazole concentration. At high concentrations, (imidazole) (NO)-6-coordinate heme is formed, and the photoinduced rebinding kinetics reveal two exponential picosecond phases ( approximately 10 and approximately 100 ps) similar to those of wild type myoglobin. At low concentrations, imidazole is displaced by the trans effect leading to a (NO)-5-coordinate heme, becoming 4 coordinate immediately after photolysis as revealed from the transient Raman spectrum. In this case, NO rebinding kinetics remain bi-exponential with no change in time constant of the fast component whose amplitude increases with respect to the 6-coordinate species. Bi-exponential NO geminate rebinding in 5 coordinate H93G-Mb is in contrast with the single-exponential process reported for nitrosylated soluble guanylate cyclase (Negrerie, M., Bouzhir, L., Martin, J. L., and Liebl, U. (2001) J. Biol. Chem. 276, 46815-46821). Thus, our data show that the iron coordination state or the heme iron out-of-plane motion are not at the origin of the bi-exponential kinetics, which depends upon the protein structure, and that the 4-coordinate state favors the fast phase of NO geminate rebinding. Consequently, the heme coordination state together with the energy barriers provided by the protein structure control the dynamics and affinity for NO-binding enzymes. PMID- 16476731 TI - Green tea polyphenols modulate insulin secretion by inhibiting glutamate dehydrogenase. AB - Insulin secretion by pancreatic beta-cells is stimulated by glucose, amino acids, and other metabolic fuels. Glutamate dehydrogenase (GDH) has been shown to play a regulatory role in this process. The importance of GDH was underscored by features of hyperinsulinemia/hyperammonemia syndrome, where a dominant mutation causes the loss of inhibition by GTP and ATP. Here we report the effects of green tea polyphenols on GDH and insulin secretion. Of the four compounds tested, epigallocatechin gallate (EGCG) and epicatechin gallate were found to inhibit GDH with nanomolar ED(50) values and were therefore found to be as potent as the physiologically important inhibitor GTP. Furthermore, we have demonstrated that EGCG inhibits BCH-stimulated insulin secretion, a process that is mediated by GDH, under conditions where GDH is no longer inhibited by high energy metabolites. EGCG does not affect glucose-stimulated insulin secretion under high energy conditions where GDH is probably fully inhibited. We have further shown that these compounds act in an allosteric manner independent of their antioxidant activity and that the beta-cell stimulatory effects are directly correlated with glutamine oxidation. These results demonstrate that EGCG, much like the activator of GDH (BCH), can facilitate dissecting the complex regulation of insulin secretion by pharmacologically modulating the effects of GDH. PMID- 16476732 TI - Apoptosis signal-regulating kinase 1 is a direct target of E2F1 and contributes to histone deacetylase inhibitor-induced apoptosis through positive feedback regulation of E2F1 apoptotic activity. AB - The oncogenic retinoblastoma protein (Rb)/E2F pathway links cellular proliferation control to apoptosis as a fail-safe mechanism to protect aberrant oncogenic transformation. We have previously shown that histone deacetylase inhibitors (HDACIs) activate the E2F1-Bim apoptotic pathway, leading to efficient cell killing in cancer cells with deregulated E2F1 activity. To identify additional gene cassettes that might contribute HDACI-induced apoptosis upon E2F1 activation, we investigated the apoptotic transcriptional network affected by HDAC inhibitor suberoylanilide hydroxamic acid (SAHA) in cancer cells with inducible E2F1. Data analysis focusing on 220 apoptosis-related genes identified apoptosis signal-regulating kinase 1 (ASK1) as one of a few genes in addition to Bim that are substantially up-regulated by SAHA upon E2F1 activation. We show that ASK1 is directly regulated by E2F1 and that prevention of ASK1 induction by RNA interference decreases SAHA-induced apoptosis. We further show that the role of ASK1 in the SAHA apoptotic response is not associated with its downstream effectors p38 or JNK. Instead, ASK1 knockdown results in reduced E2F1 transcriptional activity, leading to decreased Bim induction by SAHA. Moreover, ASK1 expression reverses the negative effect of Rb on E2F1 activity. These results indicate that ASK1 induction by E2F1 provides positive feedback regulation of E2F1 activity via Rb inhibition, which allows an efficient E2F1-Bim activation. Thus, the concomitant induction of E2F1 targets ASK1 and Bim by HDACIs warrants an effective activation of E2F1-dependent apoptosis in response to SAHA. PMID- 16476733 TI - Inhibition of S/G2 phase CDK4 reduces mitotic fidelity. AB - Cyclin-dependent kinase 4 (CDK4)/cyclin D has a key role in regulating progression through late G(1) into S phase of the cell cycle. CDK4-cyclin D complexes then persist through the latter phases of the cell cycle, although little is known about their potential roles. We have developed small molecule inhibitors that are highly selective for CDK4 and have used these to define a role for CDK4-cyclin D in G(2) phase. The addition of the CDK4 inhibitor or small interfering RNA knockdown of cyclin D3, the cyclin D partner, delayed progression through G(2) phase and mitosis. The G(2) phase delay was independent of ATM/ATR and p38 MAPK but associated with elevated Wee1. The mitotic delay was because of failure of chromosomes to migrate to the metaphase plate. However, cells eventually exited mitosis, with a resultant increase in cells with multiple or micronuclei. Inhibiting CDK4 delayed the expression of the chromosomal passenger proteins survivin and borealin, although this was unlikely to account for the mitotic phenotype. These data provide evidence for a novel function for CDK4 cyclin D3 activity in S and G(2) phase that is critical for G(2)/M progression and the fidelity of mitosis. PMID- 16476734 TI - Structural and molecular interactions of CCR5 inhibitors with CCR5. AB - We have characterized the structural and molecular interactions of CC-chemokine receptor 5 (CCR5) with three CCR5 inhibitors active against R5 human immunodeficiency virus type 1 (HIV-1) including the potent in vitro and in vivo CCR5 inhibitor aplaviroc (AVC). The data obtained with saturation binding assays and structural analyses delineated the key interactions responsible for the binding of CCR5 inhibitors with CCR5 and illustrated that their binding site is located in a predominantly lipophilic pocket in the interface of extracellular loops and within the upper transmembrane (TM) domain of CCR5. Mutations in the CCR5 binding sites of AVC decreased gp120 binding to CCR5 and the susceptibility to HIV-1 infection, although mutations in TM4 and TM5 that also decreased gp120 binding and HIV-1 infectivity had less effects on the binding of CC-chemokines, suggesting that CCR5 inhibition targeting appropriate regions might render the inhibition highly HIV-1-specific while preserving the CC chemokine-CCR5 interactions. The present data delineating residue by residue interactions of CCR5 with CCR5 inhibitors should not only help design more potent and more HIV-1 specific CCR5 inhibitors, but also give new insights into the dynamics of CC chemokine-CCR5 interactions and the mechanisms of CCR5 involvement in the process of cellular entry of HIV-1. PMID- 16476735 TI - Systematic evaluation of transcellular activities of secretory phospholipases A2. High activity of group V phospholipases A2 to induce eicosanoid biosynthesis in neighboring inflammatory cells. AB - The mechanisms by which secretory phospholipase A2 (PLA2) exerts cellular effects are not fully understood. To elucidate these mechanisms, we systematically and quantitatively assessed the activities of human group IIA, V, and X PLA2s on originating and neighboring cells using orthogonal fluorogenic substrates in various mixed cell systems. When HEK293 cells stably expressing each of these PLA2s were mixed with non-transfected HEK293 cells, group V and X PLA2s showed strong transcellular lipolytic activity, whereas group IIA PLA2 exhibited much lower transcellular activity. The transcellular activity of group V PLA2 was highly dependent on the presence of cell surface heparan sulfate proteoglycans of acceptor cells. Activation of RBL-2H3 and DLD-1 cells that express endogenous group V PLA2 led to the secretion of group V PLA2 and its transcellular action on neighboring human neutrophils and eosinophils, respectively. Similarly, activation of human bronchial epithelial cells, BEAS-2B, caused large increases in arachidonic acid and leukotriene C4 release from neighboring human eosinophils. Collectively, these studies show that group V and X PLA2s can act transcellularly on mammalian cells and suggest that group V PLA2 released from neighboring cells may function in triggering the activation of inflammatory cells under physiological conditions. PMID- 16476736 TI - Characterization of p87PIKAP, a novel regulatory subunit of phosphoinositide 3 kinase gamma that is highly expressed in heart and interacts with PDE3B. AB - Phosphoinositide 3-kinase (PI3K) gamma has been implicated in a vast array of physiological settings including the activation of different leukocyte species and the regulation of myocardial contractility. Activation of PI3Kgamma is primarily mediated by Gbetagamma subunits of heterotrimeric G proteins, which are recognized by a p101 regulatory subunit. Here, we describe the identification and characterization of a novel regulatory subunit of PI3Kgamma, which we termed p87(PIKAP) (PI3Kgamma adapter protein of 87 kDa). It is homologous to p101 in areas that we have recently shown that they mediate binding to the catalytic p110gamma subunit and to Gbetagamma. Like p101, p87(PIKAP) binds to both p110gamma and Gbetagamma and mediates activation of p110gamma downstream of G protein-coupled receptors. In contrast to p101, p87(PIKAP) is highly expressed in heart and may therefore be crucial to PI3Kgamma cardiac function. Moreover, p87(PIKAP) and p101 are both expressed in dendritic cells, macrophages, and neutrophils, raising the possibility of regulatory subunit-dependent differences in PI3Kgamma signaling within the same cell type. We further provide evidence that p87(PIKAP) physically interacts with phosphodiesterase (PDE) 3B, suggesting that p87(PIKAP) is also involved in the recently described noncatalytic scaffolding interaction of p110gamma with PDE3B. However, coexpression of PDE3B and PI3Kgamma subunits was not sufficient to reconstitute the regulatory effect of PI3Kgamma on PDE3B activity observed in heart, implying further molecules to be present in the complex regulating PDE3B in heart. PMID- 16476738 TI - Stimulation of the epithelial sodium channel (ENaC) by cAMP involves putative ERK phosphorylation sites in the C termini of the channel's beta- and gamma-subunit. AB - The mechanisms involved in the regulation of the epithelial sodium channel (ENaC) via the cAMP pathway are not yet completely understood. The aim of the present study was to investigate cAMP-mediated ENaC regulation in Xenopus laevis oocytes heterologously expressing the three subunits (alphabetagamma) of rat ENaC and to determine the ENaC regions important for mediating the stimulatory effect of cAMP. In oocytes treated for about 24 h with 1 mm 3-isobutyl-1-methylxanthine (IBMX) and 1 microm forskolin (FSK) so as to increase intracellular cAMP, the amiloride-sensitive whole cell current (DeltaI(Ami)) was on average 10-fold larger than DeltaI(Ami) in matched control oocytes. This effect on DeltaI(Ami) was paralleled by an increase in ENaC surface expression caused by a reduced rate of ENaC retrieval. In addition, IBMX/FSK also enhanced ENaC open probability from about 0.2 to 0.5. The stimulatory effect of IBMX/FSK was dependent on the presence of intact PY motifs in the C termini of the channel. Mutagenesis of putative protein kinase A and CK-2 consensus motifs in the cytosolic domains of the channel did not reveal critical sites involved in mediating the stimulatory effect of IBMX/FSK. In contrast, site-directed mutagenesis of two putative ERK consensus motifs (T613A in betaENaC and T623A in gammaENaC) largely reduced the stimulatory effect of IBMX/FSK. Phosphorylation of these ERK sites has previously been reported to enhance the interaction of ENaC and Nedd4 (Shi, H., Asher, C., Chigaev, A., Yung, Y., Reuveny, E., Seger, R., and Garty, H. (2002) J. Biol. Chem. 277, 13539-13547). Using co-expression experiments we demonstrated that mutating the two ERK sites attenuates the inhibitory effect of Nedd4-2 on ENaC currents. We conclude that an increase in intracellular cAMP favors the dephosphorylation of the two ERK sites, which reduces channel retrieval and increases P(O) by modulating ENaC/Nedd4 interaction. This defines a novel regulatory pathway likely to be relevant for cAMP-induced stimulation of ENaC in vivo. PMID- 16476737 TI - Inhibition of heme oxygenase-1 interferes with the transforming activity of the Kaposi sarcoma herpesvirus-encoded G protein-coupled receptor. AB - Heme oxygenase-1 (HO-1), the inducible enzyme responsible for the rate-limiting step in the heme catabolism, is expressed in AIDS-Kaposi sarcoma (KS) lesions. Its expression is up-regulated by the Kaposi sarcoma-associated herpesvirus (KSHV) in endothelial cells, but the mechanisms underlying KSHV-induced HO-1 expression are still unknown. In this study we investigated whether the oncogenic G protein-coupled receptor (KSHV-GPCR or vGPCR), one of the key KSHV genes involved in KS development, activated HO-1 expression. Here we show that vGPCR induces HO-1 mRNA and protein levels in fibroblasts and endothelial cells. Moreover, targeted knock-down gene expression of HO-1 by small hairpin RNA and chemical inhibition of HO-1 enzymatic activity by tin protoporphyrin IX (SnPP), impaired vGPCR-induced survival, proliferation, transformation, and vascular endothelial growth factor (VEGF)-A expression. vGPCR-expressing cells implanted in the dorsal flank of nude mice developed tumors with elevated HO-1 expression and activity. Chronic administration of SnPP to the implanted mice, under conditions that effectively blocked HO-1 activity and VEGF-A expression in the transplanted cells, strikingly reduced tumor growth, without apparent side effects. On the contrary, administration of the HO-1 inducer cobalt protoporphyrin (CoPP) further enhanced vGPCR-induced tumor growth. These data postulate HO-1 as an important mediator of vGPCR-induced tumor growth and suggest that inhibition of intratumoral HO-1 activity by SnPP may be a potential therapeutic strategy. PMID- 16476739 TI - Binding of active matrilysin to cell surface cholesterol sulfate is essential for its membrane-associated proteolytic action and induction of homotypic cell adhesion. AB - Regulation of cell surface molecules by matrix metalloproteinases (MMPs), as well as MMPs-catalyzed degradation of extracellular matrix, is important for tumor invasion and metastasis. Our previous study (Kioi, M., Yamamoto, K., Higashi, S., Koshikawa, N., Fujita, K., and Miyazaki, K. (2003) Oncogene 22, 8662-8670) demonstrated that active matrilysin specifically binds to the surface of colon cancer cells and induces notable cell aggregation due to processing of the cell membrane protein(s). Furthermore, these aggregated cells showed a dramatically enhanced metastatic potential. To elucidate the mechanism of matrilysin-induced cell aggregation, we attempted to identify the matrilysin-binding substance on the cell surface. Here, we demonstrate that cholesterol sulfate on the cell surface is a major matrilysin-binding substance. We found that active matrilysin bound to the cell membrane and cholesterol sulfate incorporated into liposomes with similar affinities. Treatment of colon cancer cells with beta-cyclodextrin significantly reduced not only matrilysin binding to the cell surface but also matrilysin-dependent proteolysis and cell aggregation. Interestingly, replenishment of cholesterol sulfate, but not cholesterol, neutralized the effects of beta-cyclodextrin. Taken together, it is likely that binding of matrilysin to cholesterol sulfate facilitates the matrilysin-catalyzed modulation of cell surface proteins, thus inducing the cancer cell aggregation. PMID- 16476740 TI - Ephrin-B1 is critical in T-cell development. AB - Eph kinases are the largest family of receptor tyrosine kinases, and their ligands, ephrins (EFNs), are also cell surface molecules. In this study, we investigated the role of EFNB1 and the Ephs it interacts with (collectively called EFNB1 receptors) in mouse T-cell development. In the thymus, CD8 single positive (SP) and CD4CD8 double positive (DP) cells expressed high levels of EFNB1 and EFNB1 receptors, whereas CD4 SP cells had moderate expression of both. Soluble EFNB1-Fc in fetal thymus organ culture caused significant subpopulation ratio skew, with increased CD4 SP and CD8 SP and decreased DP percentage, while the cellularity of the thymus remained constant. Moreover, in EFNB1-treated fetal thymus organ culture, CD117(+), CD25(+), DP, CD4 SP, and CD8 SP cells all had significantly enhanced proliferation history, according to bromodeoxyuridine uptake. In vitro culture of isolated thymocytes revealed that EFNB1-Fc on solid phase protected thymocytes from anti-CD3-induced apoptosis, with concomitant augmentation of several antiapoptotic factors, particularly in CD4 SP and CD8 SP cells; on the other hand, soluble EFNB1-Fc promoted anti-CD3-induced apoptosis, as was the case in vivo. This study reveals that EFNB1 and EFNB1 receptors are critical in thymocyte development. PMID- 16476741 TI - Stomach-specific calpain, nCL-2, localizes in mucus cells and proteolyzes the beta-subunit of coatomer complex, beta-COP. AB - Calpain is a Ca2+-regulated cytosolic protease. Mammals have 14 calpain genes, half of which are predominantly expressed in specific organ(s); the rest are expressed ubiquitously. A defect in calpains causes lethality/pathogenicity, indicating their physiological indispensability. nCL-2/calpain-8a was identified as a stomach-specific calpain, whose physiological functions are unclear. To elucidate these, we characterized nCL-2 in detail. Unexpectedly, nCL-2 was localized strictly to the surface mucus cells in the gastric epithelium and the mucus-secreting goblet cells in the duodenum. Yeast two-hybrid screening identified several nCL-2-interacting molecules. Of these, the beta-subunit of coatomer complex (beta-COP) occurs in the stomach pit cells and is proteolyzed by nCL-2 in vitro. Furthermore, beta-COP and nCL-2 co-expressed in COS7 cells co localized in the Golgi, and Ca2+-ionophore stimulation caused the proteolysis of beta-COP near the linker region, resulting in the dissociation of beta-COP from the Golgi. These results strongly suggest novel functions for nCL-2 that involve the membrane trafficking of mucus cells via interactions with coat protein. PMID- 16476742 TI - Phosphorylation of beta-catenin by cyclic AMP-dependent protein kinase. AB - Beta-catenin is a signaling molecule that promotes cell proliferation by the induction of gene transcription through the activation of T-cell factor (TCF)/lymphoid enhancer factor (LEF) transcription factors. The canonical mechanism of the regulation of beta-catenin involves its phosphorylation by casein kinase 1 at the Ser-45 site and by glycogen synthase kinase 3 (GSK3) at the Thr-41, Ser-37, and Ser-33 sites. This phosphorylation targets beta-catenin to ubiquitination and degradation by the proteasome system. Mitogenic factors promote beta-catenin signaling through the inhibition of GSK3, resulting in reduced beta-catenin phosphorylation, its stabilization, and subsequent accumulation in the nucleus, where it stimulates TCF/LEF-dependent gene transcription. In the present study, we have shown that (i) beta-catenin can be phosphorylated by protein kinase A (PKA) in vitro and in intact cells at two novel sites, Ser-552 and Ser-675; (ii) phosphorylation by PKA promotes the transcriptional activity (TCF/LEF transactivation) of beta-catenin; (iii) mutation of Ser-675 attenuates the promoting effect of PKA; (iv) phosphorylation by PKA does not affect the GSK3-dependent phosphorylation of beta-catenin, its stability, or intracellular localization; and (v) phosphorylation at the Ser-675 site promotes the binding of beta-catenin to its transcriptional coactivator, CREB-binding protein. In conclusion, this study identifies a novel, noncanonical mechanism of modulation of beta-catenin signaling through direct phosphorylation of beta-catenin by PKA, promoting its interaction with CREB-binding protein. PMID- 16476743 TI - Targeted disruption of Gb3/CD77 synthase gene resulted in the complete deletion of globo-series glycosphingolipids and loss of sensitivity to verotoxins. AB - To examine whether globotriaosylceramide (Gb3/CD77) is a receptor for verotoxins (VTs) in vivo, sensitivity of Gb3/CD77 synthase null mutant mice to VT-2 and VT-1 was analyzed. Although wild-type mice died after administration of 0.02 microg of VT-2 or 1.0 microg of VT-1, the mutant mice showed no reaction to doses as much as 100 times that administered to wild types. Expression analysis of Gb3/CD77 in mouse tissues with antibody revealed that low, but definite, levels of Gb3/CD77 were expressed in the microvascular endothelial cells of the brain cortex and pia mater and in renal tubular capillaries. Corresponding to the Gb3/CD77 expression, tissue damage with edema, congestion, and cytopathic changes was observed, indicating that Gb3/CD77 (and its derivatives) exclusively function as a receptor for VTs in vivo. The lethal kinetics were similar regardless of lipopolysaccharide elimination in VT preparation, suggesting that basal Gb3/CD77 levels are sufficient for lethal effects of VTs. PMID- 16476744 TI - Removing organs "just in case"--is prophylactic removal of the ovaries a good thing? PMID- 16476745 TI - What same sex civil partnerships may mean for health. AB - A growing number of countries have introduced a form of marriage or civil partnership registration for same sex couples. Marriage confers health benefits on heterosexual men and women and similar benefits could arise from same sex civil unions. The authors argue that legal and social recognition of same sex relationships may reduce discrimination, increase the stability of same sex relationships, and lead to better physical and mental health for gay and lesbian people. PMID- 16476746 TI - Infectious diseases epidemiology. AB - In this glossary the authors have reviewed old and new terms contemporarily used in the infectious disease epidemiology. Many of these concepts were established throughout the 19th century and at the beginning of the 20th century (classic terms), however, the meanings of old terms have been revised and new terms are continually being added. This glossary has therefore reviewed the classic and the more recently established terminology defining the most relevant terms contemporarily used in this field. PMID- 16476747 TI - Use of health impact assessment in incorporating health considerations in decision making. AB - STUDY AIM: The aim of this project is to identify from a range of sources the factors associated with the success of a health impact assessment (HIA) in integrating health considerations into the final decision and implementation of a planned policy, programme, or project. DESIGN: Three methods were adopted: (a) a review of HIA case studies; (b) a review of commentaries, reviews and discussion papers relating to HIA and decision making; and (c) an email survey of a purposive sample of HIA academics, HIA practitioners, and policymakers. Information was captured on the following characteristics: information on the year undertaken; geopolitical level; setting; sector; HIA type; methods and techniques used; identification of assessors. MAIN RESULTS: Two groups of factors were identified relating to the decision making environment and to the technical conduct of the HIA. With regard to the environment, striking a balance between decision maker ownership and HIA credibility; organisational, statutory and policy commitment to HIA, and the provision of realistic, non-controversial recommendations were cited as enablers to the integration of HIA findings into the decision making process. Barriers included a lack of knowledge of the policymaking environment by those conducting HIA. Regarding factors relating to the conduct of the HIA: use of a consistent methodological approach; inclusion of empirical evidence on health impacts; timing of the HIA congruent with the decision making process; involvement of expert HIA assessors; and shaping of recommendations to reflect organisational priorities were cited as enablers while lack of a standardised methodology; lack of resources and use of jargon were cited as barriers. CONCLUSIONS: The findings emphasise the importance of considering the politico-administrative environment in which HIA operates. The extent to which HIA fits the requirements of organisations and decision makers may be as important as the technical methods adopted to undertake it. PMID- 16476749 TI - Socioeconomic deprivation, travel distance, location of service, and uptake of breast cancer screening in North Derbyshire, UK. AB - BACKGROUND AND AIM: This study examined the association between socioeconomic deprivation, travel distance, urban-rural status, location and type of screening unit, and breast screening uptake. Screening was provided at 13 locations--1 fixed and 12 mobile (3 at non-health locations). METHODS: The study examined data from 1998 to 2001 for 34 868 women aged 50-64 years, calculated road travel distance, used 1991 enumeration district level Townsend socioeconomic deprivation scores, and a ward level urban-rural classification. RESULTS: Odds of attendance for screening decreased with increasing socioeconomic deprivation, with an adjusted odds ratio of 0.64 (95%CI 0.59 to 0.70) in the most deprived relative to the least deprived category. 87% of women lived within 8 km of their screening location. The odds ratio for a 10 km increase in distance was 0.87 (95%CI 0.79 to 0.95). The odds ratios were 1.18 (95%CI 1.08 to 1.28) for screening at a non health relative to a health location, 1.00 (95%CI 0.94 to 1.07) for the fixed site relative to the mobile unit and 1.00 (95%CI 0.91 to 1.09) for mainly rural relative to mainly urban areas. CONCLUSIONS: Socioeconomic inequality in breast screening uptake seems to persist in an established service. There was a small decrease with increasing distance, no difference between fixed and mobile units, and no difference between urban and rural areas but uptake seemed to be higher at non-health sites. Further work is needed to identify effective methods of decreasing socioeconomic inequalities in uptake and to confirm if non-health locations are associated with higher screening uptake. PMID- 16476748 TI - Effect of area poverty rate on cancer screening across US communities. AB - STUDY OBJECTIVE: To analyse the contextual effect of area poverty rate on never having been screened for breast, cervical, and colorectal cancer by (1) describing the extent of the variation in screening behaviours among 98 US metropolitan areas; (2) determining if the variation in lack of screening can be explained by differences in the characteristics of the persons who resided in these areas; and (3) determining if living in a metropolitan area with a higher poverty rate increased the likelihood of never having been screened for cancer over and above individual characteristics. DESIGN: Cross sectional survey using data from the 2002 Behavioral Risk Factor Surveillance System. Multilevel logistic regression included both individual level factors as well as area poverty rate. SETTING: Ninety eight areas across the USA. PARTICIPANTS: Over 118 000 persons residing in 98 areas; a sample aimed at estimating 48.3% of the US population age 18 or older. MAIN RESULTS: After adjustment for individual level factors, increasing area level poverty rate (per 5%) remained associated with never having had a mammogram (odds ratio (OR) = 1.28, 95% confidence interval (CI): 1.03 to 1.37); clinical breast examination (OR = 1.28, 95% CI: 1.11 to 1.48), colonoscopy/sigmoidoscopy (OR = 1.10, 95% CI: 1.01 to 1.19), and a faecal occult blood test (OR = 1.19, 95% CI: 1.12 to 1.27). Poverty rate was not independently associated with never having had a Pap smear (OR = 1.12; 95% CI: 0.90 to 1.41). The size of the variance among metropolitan or micropolitan statistical areas (MMSAs) varied by type of screening test, with intraclass correlation coefficients ranging from 4.9% (never having had a Pap smear) to 1.2% (never having had a colonoscopy/sigmoidoscopy). CONCLUSIONS: Area poverty rate was independently associated with never having been screened for breast and colorectal cancer, but not cervical cancer. The size of the variance among MMSAs was modest at best. PMID- 16476750 TI - The polypill: at what price would it become cost effective? AB - INTRODUCTION: A promising concept in cardiovascular disease prevention (the polypill) was introduced in 2003. Although the polypill may seem as an effective intervention, data on its costs and cost effectiveness remain unknown. The aim of this study was to determine the maximum price of the polypill for it to be a cost effective alternative in the primary prevention of cardiovascular disease. METHODS: Data on the hypothetical effects of the polypill were taken from the literature. Using data from the Framingham heart study and the Framingham offspring study, life tables were built to model the assumed benefits of the polypill. Using a third party payer perspective and a 10 years time horizon, the authors calculated what should be the maximum drug cost of the polypill for it to be cost effective (using a 20,000 euro/year of life saved threshold) in the primary prevention of cardiovascular disease among populations at different levels of absolute risk of coronary heart disease and age. RESULTS: To be cost effective among populations at levels of 10 year coronary heart disease risk over 20% (high risk), the annual cost of medication for the polypill therapy should be no more than 302 euro or 410 euro for men at age 50 and 60 years respectively. For cost effective prevention in populations at levels of coronary heart disease risk between 10% and 20% the costs should be two to three times lower. CONCLUSION: Although the polypill could theoretically be a highly effective intervention, the costs of the medication could be its caveat for implementation in the primary prevention of cardiovascular disease. PMID- 16476751 TI - Changes in socioeconomic inequalities in census measures of health in England and Wales, 1991-2001. AB - STUDY OBJECTIVES: To investigate changes in socioeconomic inequalities in census measures of health in England and Wales between 1991 and 2001. DESIGN: Indirect standardisation was used to calculate age standardised rates of limiting long term illness and permanent sickness in men and women in all residential wards in England and Wales in 1991 and 2001. The socioeconomic position of each ward was determined using Townsend deprivation scores. SETTING: All residential wards in England and Wales in 1991 and 2001. PARTICIPANTS: All people aged 16-65 who provided census information in the 1991 or 2001 censuses. MAIN RESULTS: There was strong evidence that Townsend deprivation score quintile could predict both logged standardised permanent sickness rate and logged standardised limiting long term illness rate. There was evidence that socioeconomic inequalities in standardised limiting long term illness rates decreased between 1991 and 2001 in both men and women and that socioeconomic inequalities in standardised permanent sickness rates decreased in women but increased in men between 1991 and 2001. CONCLUSIONS: As permanent sickness rates seem to reflect labour market accessibility, this study may have found evidence that socioeconomic inequalities in self reported morbidity decreased but inequalities in labour market participation in men increased between 1991 and 2001. PMID- 16476752 TI - Sociodemographic predictors of antenatal and postpartum depressive symptoms among women in a medical group practice. AB - OBJECTIVE: Data are scarce regarding the sociodemographic predictors of antenatal and postpartum depression. This study investigated whether race/ethnicity, age, finances, and partnership status were associated with antenatal and postpartum depressive symptoms. SETTING: 1662 participants in Project Viva, a US cohort study. DESIGN: Mothers indicated mid-pregnancy and six month postpartum depressive symptoms on the Edinburgh postpartum depression scale (EPDS). Associations of sociodemographic factors with odds of scoring >12 on the EPDS were estimated. MAIN RESULTS: The prevalence of depressive symptoms was 9% at mid pregnancy and 8% postpartum. Black and Hispanic mothers had a higher prevalence of depressive symptoms compared with non-Hispanic white mothers. These associations were explained by lower income, financial hardship, and higher incidence of poor pregnancy outcome among minority women. Young maternal age was associated with greater risk of antenatal and postpartum depressive symptoms, largely attributable to the prevalence of financial hardship, unwanted pregnancy, and lack of a partner. The strongest risk factor for antenatal depressive symptoms was a history of depression (OR = 4.07; 95% CI 3.76, 4.40), and the strongest risk for postpartum depressive symptoms was depressive symptoms during pregnancy (6.78; 4.07, 11.31) or a history of depression before pregnancy (3.82; 2.31, 6.31). CONCLUSIONS: Financial hardship and unwanted pregnancy are associated with antenatal and postpartum depressive symptoms. Women with a history of depression and those with poor pregnancy outcomes are especially vulnerable to depressive symptoms during the childbearing year. Once these factors are taken in account, minority mothers have the same risk of antenatal and postpartum depressive symptoms as white mothers. PMID- 16476753 TI - Socioeconomic lifecourse influences on women's smoking status in early adulthood. AB - STUDY OBJECTIVE: To incorporate women's domestic trajectories and circumstances into analyses of the socioeconomic influences on women's smoking status (current and former smoking) in early adulthood. DESIGN: Cross sectional survey. SETTING: Southampton, UK. PARTICIPANTS: 8437 women aged 25-34 recruited from 1998-2002 via patient lists of general practices. MAIN RESULTS: Domestic lifecourse factors contributed to the odds of being a current smoker and former smoker in models that included conventional measures of the socioeconomic lifecourse. Early motherhood, non-cohabitation, and lone motherhood increased the odds of smoking; early motherhood and non-cohabitation reduced the odds of former smoking. For example, relative to childless women, odds ratios (OR) for women who had become mothers <20 years were 1.71 for smoking and 0.76 for former smoking. The effects of education and current SEP remained strong with the inclusion of childbearing and cohabitation variables for both outcomes. For instance, compared with women in education to age > or =22, the odds ratio for smoking for those leaving school < or =16 was 3.37 and for former smoking was 0.42. CONCLUSIONS: Both the conventionally measured socioeconomic lifecourse and the domestic lifecourse contributed separately to the odds of smoking and former smoking, suggesting that lifecourse analyses should incorporate women's domestic circumstances as an important pathway of influence on their smoking status in early adulthood. PMID- 16476754 TI - Children's exposure to nitrogen dioxide in Sweden: investigating environmental injustice in an egalitarian country. AB - STUDY OBJECTIVE: Prior studies have shown that children are particularly sensitive to air pollution. This study examined whether children of low socioeconomic status suffered greater exposure to outdoor nitrogen dioxide than more affluent ones, both at their place of residence and at school, in a country with widespread state intervention for social equity. DESIGN: Local scale data on outdoor nitrogen dioxide obtained from a validated air pollution model were analysed, along with all school children accurately geocoded to their building of residence and school. PARTICIPANTS: All 29,133 children in grades one through nine (aged 7 to 15 years) residing and attending school in Malmo, Sweden, in 2001. MAIN RESULTS: Defining the socioeconomic status of children according to the mean income in their residential building, the spatial scan statistic technique allowed the authors to identify eight statistically significant clusters of low socioeconomic status children, all of which were located in the most polluted areas of Malmo. Four clusters of high socioeconomic status children were found, all of them located in the least polluted areas. The neighbourhood socioeconomic status better predicted the nitrogen dioxide exposure of children than the socioeconomic status of their building of residence. Exposure to nitrogen dioxide at the place of residence and school of attendance regularly increased as the socioeconomic status of a child's neighbourhood of residence decreased. CONCLUSIONS: Evidence of environmental injustice was found, even in a country noted for its egalitarian welfare state. Enforcement of environmental regulations may be necessary to achieve a higher level of environmental equity. PMID- 16476755 TI - Prevalence of arsenic exposure and skin lesions. A population based survey in Matlab, Bangladesh. AB - STUDY OBJECTIVE: To assess prevalence of arsenic exposure through drinking water and skin lesions, and their variation by geographical area, age, sex, and socioeconomic conditions. DESIGN, SETTING, AND PARTICIPANTS: Skin lesion cases were identified by screening the entire population above 4 years of age (n = 166,934) living in Matlab, a rural area in Bangladesh, during January 2002 and August 2003. The process of case identification involved initial skin examinations in the field, followed by verification by physicians in a clinic, and final confirmation by two independent experts reviewing photographs. The tubewell water arsenic concentrations (n = 13,286) were analysed by atomic absorption spectrometry. Drinking water history since 1970 was obtained for each person. Exposure information was constructed using drinking water histories and data on water arsenic concentrations. MAIN RESULTS: The arsenic concentrations ranged from <1 to 3644 microg/l, and more than 70% of functioning tubewells exceeded the World Health Organisation guideline of 10 microg/l. Arsenic exposure had increased steadily from 1970s to the late 1990s, afterwards a decrease could be noted. In total, 504 skin lesions cases were identified, and the overall crude prevalence was 3/1000. Women had significantly higher cumulative exposure to arsenic, while men had significantly higher prevalence of skin lesions (SMR 158, 95% CI 133 to 188). The highest prevalence occurred in 35-44 age groups for both sexes. Arsenic exposure and skin lesions had a positive association with socioeconomic groups and achieved educational level. CONCLUSIONS: The result showed sex, age, and socioeconomic differentials in both exposure and skin lesions. Findings clearly showed the urgency of effective arsenic mitigation activities. PMID- 16476756 TI - Injury mortality among ethnic minority groups in the Netherlands. AB - OBJECTIVES: To prepare a comprehensive overview of ethnic differences in injury related mortality in the Netherlands and to determine the role of area income and urbanisation degree. METHODS: Data for the period 1995-2000 were obtained from the population and cause of death registries. Injury related death rates were compared for persons from Turkish, Moroccans, Surinamese, and Antillean/Aruban origin with rates for the native Dutch population. RESULTS: Compared with the native Dutch population, all ethnic minorities combined had an increased mortality for all injuries together (RR = 1.29). Ethnic minorities experienced a significantly higher risk of death from pedestrian accidents (RR = 1.87), drowning (RR = 2.58), poisoning (RR = 1.76), fire and scalds (RR = 1.95), and homicide (RR = 3.24). Mortality for cyclists (RR = 0.53) and motorcycle drivers (RR = 0.47) was significantly lower among ethnic minorities compared with the native Dutch. Adjustment for area income and urbanisation decreased the mortality risk difference for most of the non-traffic injuries, but showed a difference in risk for car driver and passenger accidents (RR = 1.37). Compared with the native Dutch inhabitants, Surinamese and Antillean/Aruban population had a higher risk of total injury mortality (RR = 1.33 and 1.53 respectively), while Turkish and Moroccans had increased risk only for selected conditions. Inequalities in injury mortality were the highest among children and young adults, but persisted in the age group above 50 years old. CONCLUSION: Ethnic differences in injury mortality in the Netherlands strongly depended on type of injury, ethnic group, sex, and age. Policies should be aimed at the prevention of high risk injuries among the most vulnerable ages and ethnic groups. PMID- 16476757 TI - Respiratory diseases and pesticide exposure: a case-control study in Lebanon. AB - STUDY OBJECTIVE: To evaluate the odds of being exposed to pesticides in asthmatic adults. DESIGN: A case-control study was performed in Lebanon. SETTING: People were approached when consulting physicians as outpatients. PATIENTS: Asthmatic patients and non-asthmatic controls in several Lebanese hospitals were interviewed. MAIN RESULTS: The study included 407 subjects from 10 medical centres. Any exposure to pesticides was associated to asthma (OR = 2.11 (1.47 to 3.02); p<10(-4)). Occupational use presented the highest association (OR = 4.98 (1.07 to 23.28); p = 0.02), followed by regional exposure (OR 3.51 (2.11 to 5.85); p<10(-4)). Results were confirmed by multivariate analysis, particularly for regional exposure (OR(a) = 2.78; p = 0.02) and house exposure (OR(a) = 2.17; p = 0.001). CONCLUSIONS: Results are comparable to those found in other studies; especially for occupational exposure. Pesticides toxicological effects may explain chronic respiratory symptoms and asthma associations found with all exposure types. Pesticide exposure was associated with asthma in Lebanese adults. PMID- 16476758 TI - Myocardial infarction deaths after high level exposure to particulate matter. AB - STUDY OBJECTIVE: To examine the short term effects of raised concentrations of air pollutants on myocardial infarction deaths, the authors measured if incidence rate ratios increased after the concentration of suspended particulate matter (SPM) reached various critical values and were sustained for various periods of time. DESIGN: Retrospective analysis of a database, which contained hourly SPM concentrations and myocardial infarction deaths. METHODS: The rate of death (events/1000 hours) attributable to myocardial infarction was compared among SPM categories. SPM categories were classified by five different critical values that measured high SPM levels, and by the hours of exposure (exposure window) after reaching the critical SPM level. MAIN OUTCOME MEASURES: The adjusted incidence rate ratios in the various SPM categories were compared with the reference category (0-99 microg/m(3)) to determine the incidence rate ratio. SETTING: Tokyo Metropolitan area, Japan. Patients/ PARTICIPANTS: 14,950 people who died of myocardial infarction in the Tokyo Metropolitan area from 1990 to 1994. MAIN RESULTS: When the exposure window was one hour, the adjusted incidence rate ratio in each category were 1.13 (95% CI: 1.07 to 1.20) in the 100-149 microg/m(3) category, 1.17-1.24 in the intermediate categories, and 1.40 (1.00 to 1.97) in the highest (300 microg/m(3) over) category. When the exposure window was one to six hours, increased rate ratios were seen in the highest category (1.17 (four hours) to 1.40 (one hour)). Gradual increases in the incidence rate ratio were seen as the SPM category increased when the exposure window was less than six hours. CONCLUSION: An increased rate ratio of myocardial infarction deaths was seen within a few hours after reaching a high concentration of SPM. When the exposure window was less than six hours, there was a gradual increase in the incidence rate ratio as the SPM concentration increased. PMID- 16476759 TI - Socioeconomic position and sleep quantity in UK adults. AB - STUDY OBJECTIVE: To investigate the association between markers of socioeconomic position (years of full time education, access to a car or van, and occupational social class) and mid-range sleep quantity (6.5 to 8.5 hours). DATA SOURCE: Office of National Statistics Omnibus Survey, May 1999. PARTICIPANTS: 3000 households were randomly selected from the UK postcode address file and one person aged over 16 years from each household was randomly selected and invited to take part in the survey. All respondents aged 25 and over (n = 1473) were included in the analyses where appropriate data were available. MAIN RESULTS: Total sleep quantity was greater in more deprived women as measured by years in full time education. There was no linear association between total sleep quantity and any of the markers of socioeconomic position in men. Mid-range sleep quantity was more common in more educated women. Similar, non-significant, trends were seen in men. CONCLUSION: There is little evidence that more socioeconomically deprived people obtain less sleep than more advantaged ones--indeed, the reverse may be true--but some evidence that more advantaged women are more likely to report mid-range sleep. Any hypothesis implicating sleep in socioeconomic inequalities in health should take into account mid-range, rather than total, sleep quantity. Further work should focus both on mid-range, or "healthy" sleep, quantity as well as sleep quality. PMID- 16476760 TI - Leisure time exercise and personal circumstances in the working age population: longitudinal analysis of the British household panel survey. AB - OBJECTIVES: Investigate the impact of social, economic, and family circumstances on participation in weekly leisure time exercise. DESIGN: Longitudinal regression analysis of the British household panel survey. PARTICIPANTS: 9473 people (4521 men and 4952 women) giving 27,881 person years of responses across eight years and four survey waves. MAIN RESULTS: There was considerable variation among people in regular exercise participation over time. Having children was associated with a reduced likelihood of exercise for both men and women, although there were sex differences in this association according to the age of the youngest child. For both men and women working long hours was associated with a reduced likelihood of exercise, as was having a lower grade job. Retired men and women were more likely to exercise, as were those who attended a fee paying school. There was no strong independent association between household income and exercise. CONCLUSIONS: For most people, participation in leisure time exercise "comes and goes" rather than being something they always or never do. Those with time pressures from work or domestic life are less likely to participate in leisure time physical activity. There are important sex differences in the impact of having children, with women experiencing longer term detrimental effects. Working long hours reduces leisure time exercise participation. Opportunities for physical activity as part of our daily working routines should be increased. PMID- 16476761 TI - Skinfold thickness, body mass index, and fatal coronary heart disease: 30 year follow up of the Northwick Park heart study. AB - STUDY OBJECTIVE: To examine the effect of baseline body mass index (BMI) and skinfold thickness (ST) on fatal coronary heart disease (CHD) and all cause mortality after 30 years of follow up. DESIGN: Prospective cohort study. SETTING: Northwick Park heart study (NPHS) designed to investigate the role of haemostatic variables on CHD. PARTICIPANTS: 1511 men and 691 women enrolled in NPHS aged 40 to 64 years at entry. MAIN RESULTS: Baseline BMI (kg/m(2)) and forearm, triceps, subscapular, and suprailiac skinfolds ST (mm) were measured. Cox regression was used to calculate hazard ratios for fatal CHD and total mortality for each standard deviation unit increase in obesity adjusting for age, smoking status, total cholesterol, systolic blood pressure, fibrinogen, and factor VII activity. Subjects experienced 250 fatal CHDs and 819 all cause deaths over 30 years (median: 26 years; IQR: 22-28 years). Among men, only BMI (RR = 1.29, 95%CI = 1.12 to 1.49) significantly increased the risk of fatal CHD. Among women, BMI (RR = 1.48, 95%CI = 1.07 to 2.06), as well as, subscapular (RR = 1.65, 95%CI = 1.19 to 2.30), forearm (RR = 1.46, 95%CI = 1.08 to 1.97), and triceps (RR = 1.63, 95%CI = 1.12 to 2.39) skinfolds were predictive of fatal CHD. None of the estimates for all cause mortality were significant except for subscapular skinfold in women (RR = 1.20, 95%CI = 1.02 to 1.42). There was no evidence of interaction between obesity and sex for fatal CHD or all cause death. The effect of obesity on fatal CHD or all cause deaths does not seem to be mediated substantially by cholesterol, systolic blood pressure, or haemostatic variables. CONCLUSIONS: BMI is an important risk factor for fatal CHD where its prognostic significance remains after up to 30 years of follow up. PMID- 16476762 TI - Serum osteoprotegerin and RANKL levels in chronic alcoholic liver disease. AB - OBJECTIVES: Osteoprotegerin (OPG) is a decoy receptor that binds RANK-ligand (RANKL) and prevents osteoclast activation. Oestrogens, androgens, corticosteroids, parathyroid hormone (PTH), vitamin D, and several cytokines exert their effects on bone modulating the OPG/RANKL system. Since these substances become altered in chronic alcoholic liver disease, we investigated the OPG/RANKL system in alcoholic liver disease, its relation with bone mineral density (BMD) and with several hormones and cytokines. METHODS: Serum OPG, RANKL, C-terminal cross-linking telopeptide of type 1 collagen, osteocalcin, insulin like growth factor 1 (IGF-1), 1,25 dihydroxyvitamin D, IL-6, tumour necrosis factor (TNF)-alpha, PTH, estradiol, free testosterone and corticosterone were measured in 77 male alcoholic patients, 25 of them cirrhotics. All these patients underwent assessment of BMD at lumbar spine and left hip by a Hologic QDR-2000 (Waltham, MA) bone densitometer. Nineteen non-drinkers male sanitary workers of similar age served as controls. RESULTS: Serum OPG levels were higher in patients (12.66 +/- 6.44 pmol/l) than in controls (6.59 +/- 1.58 pml/l, P < 0.005), especially in cirrhotics (15.97 +/- 7.03 pmol/l) vs non-cirrhotics (10.96 +/- 5.45 pmol/l, P < 0.001). Patients also showed higher telopeptide levels (0.60 +/- 0.36 vs 0.20 +/- 0.10 nmol/100 ml, P < 0.001), less IGF-1 [median = 192, interquartile range (IQR) = 46.7-175.99 ng/ml vs 150, IQR = 118.8-239.4 ng/ml, P < 0.001], vitamin D (25.5, IQR = 18.25-35 pg/ml vs 77.89, IQR = 57.48-98.53 pg/ml, P < 0.001) and osteocalcin (1.8, IQR = 1-3.6 ng/ml vs 6.04, IQR = 4.63 8.20 ng/ml, P < 0.001) than controls, but no differences in PTH and RANKL. Patients also showed lower Z-scores than controls at trochanter (-0.36 +/- 1.10 vs 0.26 +/- 0.87 in controls, P = 0.026), intertrochantereal area (-0.56 +/- 1.16 vs 0.46 +/- 1.01, P = 0.001), and total hip (-0.44 +/- 1.12 vs 0.42 +/- 1, P = 0.003). TNF-alpha levels were higher in patients (7.40, IQR = 4.30-17.80 pg/ml) than in controls (5.10, IQR = 4.40-8 pg/ml, P = 0.009), especially in cirrhotics (median = 13.90, IR = 6.10-21.10 pg/ml). OPG levels showed strong correlations with TNF-alpha (rho = 0.57, P < 0.001) and IL-6 (r = 0.62, P < 0.001), but not with BMD. Estradiol levels (31.83 +/- 13.11 pg/ml) were higher and free testosterone lower (13.62 +/- 11.96 pg/ml) in patients than in controls (20.36 +/ 3.08 and 18.19 +/- 4.68 pg/ml, respectively, P < 0.001 in both cases). CONCLUSION: OPG is raised in alcoholics, especially in cirrhotics, showing no relationship with decreased BMD. Also, raised TNF and IL-6 were observed, and were strongly, directly related with OPG levels. Since TNF and IL-6 enhance bone resorption, their relation with OPG suggests a protective effect of raised OPG on bone loss. PMID- 16476763 TI - Intermittent presentations of ethanol sipper tube induce ethanol drinking in rats. AB - AIMS: Intermittent presentations of the ethanol sipper have been reported to induce more ethanol drinking in rats than when the ethanol sipper was continuously available during the session. This intermittent sipper effect was observed in a social drinking situation, in which subjects experienced intermittent opportunities to interact briefly with a conspecific rat. The objective of this study was to evaluate the effects of the intermittent sipper procedure in situations providing for intermittent presentations of food, and, in addition, in situations that do not provide for intermittent presentations of another rewarding event. METHODS: Four groups of male Long-Evans hooded rats, arranged in a 2 x 2 factorial design with two levels of Sipper Procedure (Intermittent vs Continuous) and two levels of Food procedure (Food vs No Food), were trained in drinking chambers. During each daily session, Intermittent Sipper groups received access to the ethanol sipper during each of 25 trials of 10 s each, while Continuous Sipper groups had access to the ethanol sipper during the entire session (approximately 30 min). During each session, Food groups received 25 presentations of food pellets while No Food groups received no food pellets. Ethanol concentrations in the sipper [3, 4, 6, 8, and 10% (vol./vol.)] increased across sessions. RESULTS: More rapid escalation of ethanol intake was observed in the Intermittent Sipper groups than in the Continuous Sipper groups, and this effect was observed in both the Food and No Food conditions (P's < 0.05), which did not differ from one another. CONCLUSIONS: Intermittent Sipper procedures provide less access to the ethanol sipper, yet induced more ethanol drinking than Continuous Sipper procedures. The intermittent sipper effect is not dependent on presentations of food. Implications for schedule-induced polydipsia and Pavlovian autoshaping are discussed. PMID- 16476764 TI - Predictors of relapse to harmful alcohol after orthotopic liver transplantation. AB - BACKGROUND: End-stage alcoholic liver disease (ALD) is a common indication for liver transplantation. Outcomes may be limited by return to harmful drinking. Previous studies have identified few predictors of drinking relapse. AIM: This study examined novel postulated predictors of relapse to drinking. METHOD: The case notes of all patients transplanted for ALD at the Royal Prince Alfred Hospital from 1987-2004 were reviewed. Pre-transplant characteristics were rated by a psychiatrist independent of the transplant team, blind to the outcome. Outcomes were rated by a second independent alcohol treatment specialist also blind to the pre-transplant ratings. RESULTS: Of 100 patients, 6 died before discharge from hospital, 4 had <6 months follow-up, 18 relapsed to harmful drinking, 10 drank below harmful levels, and 62 remained abstinent after a mean of 5.6 years follow-up. Univariate analyses identified six potential pre transplant predictors of return to harmful drinking. These were a diagnosis of mental illness (of which all cases were of depression), the lack of a stable partner, grams per day consumed in the years before assessment for transplant, reliance on 'family or friends' for post-transplant support, tobacco consumption at time of assessment, and lack of insight into the alcohol aetiology. Duration of pre-transplant abstinence and social class by occupation did not predict relapse. A multivariate model based on the above characteristics correctly predicted 89% of the outcomes. CONCLUSION: A model based on readily defined behaviours and psychosocial factors predicted relapse to harmful drinking after transplant for ALD. This model may improve assessment and post-transplant management of patients with advanced ALD. PMID- 16476765 TI - Isoprenoids determine Th1/Th2 fate in pathogenic T cells, providing a mechanism of modulation of autoimmunity by atorvastatin. AB - 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase is a critical enzyme in the mevalonate pathway that regulates the biosynthesis of cholesterol as well as isoprenoids that mediate the membrane association of certain GTPases. Blockade of this enzyme by atorvastatin (AT) inhibits the destructive proinflammatory T helper cell (Th)1 response during experimental autoimmune encephalomyelitis and may be beneficial in the treatment of multiple sclerosis and other Th1-mediated autoimmune diseases. Here we present evidence linking specific isoprenoid intermediates of the mevalonate pathway to signaling pathways that regulate T cell autoimmunity. We demonstrate that the isoprenoid geranylgeranyl pyrophosphate (GGPP) mediates proliferation, whereas both GGPP and its precursor, farnesyl-PP, regulate the Th1 differentiation of myelin-reactive T cells. Depletion of these isoprenoid intermediates in vivo via oral AT administration hindered these T cell responses by decreasing geranylgeranylated RhoA and farnesylated Ras at the plasma membrane. This was associated with reduced extracellular signal-regulated kinase (ERK) and p38 phosphorylation and DNA binding of their cotarget c-fos in response to T cell receptor activation. Inhibition of ERK and p38 mimicked the effects of AT and induced a Th2 cytokine shift. Thus, by connecting isoprenoid availability to regulation of Th1/Th2 fate, we have elucidated a mechanism by which AT may suppress Th1-mediated central nervous system autoimmune disease. PMID- 16476766 TI - Complete differentiation of CD8+ T cells activated locally within the transplanted liver. AB - The transplanted liver elicits systemic tolerance, and the underlying mechanism may also account for the persistence of liver infections, such as malaria and viral hepatitis. These phenomena have led to the hypothesis that antigen presentation within the liver is abortive, leading to T cell tolerance or apoptosis. Here we test this hypothesis in an optimized orthotopic liver transplantation model. In direct contradiction to this model, the liver itself induces full CD8+ T cell activation and differentiation. The effects of microchimerism were neutralized by bone marrow transplantation in the liver donor, and the lack of liver-derived antigen-presenting cells was documented by eight-color flow cytometry and by sensitive functional assays. We conclude that local antigen presentation cannot explain liver tolerance. On the contrary, the liver may be an excellent priming site for naive CD8+ T cells. PMID- 16476767 TI - Impaired response to Listeria in H2-M3-deficient mice reveals a nonredundant role of MHC class Ib-specific T cells in host defense. AB - The major histocompatibility complex (MHC) class Ib molecule H2-M3 primes the rapid expansion of CD8+ T cells by presenting N-formylated bacterial peptides. However, the significance of H2-M3-restricted T cells in host defense against bacteria is unclear. We generated H2-M3-deficient mice to investigate the role of H2-M3 in immunity against Listeria monocytogenes (LM), a model intracellular bacterial pathogen. H2-M3-deficient mice are impaired in early bacterial clearance during primary infection, with diminished LM-specific CD8+ T cell responses and compromised innate immune functions. Although H2-M3-restricted CD8+ T cells constitute a significant proportion of the anti-listerial CD8+ T cell repertoire, the kinetics and magnitude of MHC class Ia-restricted T cell responses are not altered in H2-M3-deficient mice. The fact that MHC class Ia restricted responses cannot compensate for the H2-M3-mediated immunity suggests a nonredundant role of H2-M3 in the protective immunity against LM. Thus, the early H2-M3-restricted response temporally bridges the gap between innate and adaptive immune responses, subsequently affecting the function of both branches of the immune system. PMID- 16476768 TI - Human antibodies induce arthritis in mice deficient in the low-affinity inhibitory IgG receptor Fc gamma RIIB. AB - Rheumatoid arthritis (RA) is a complex autoimmune disease with a poorly understood pathogenesis. The disease is associated with polyclonal B cell activation and the production of autoantibodies (autoAbs), but there is a longstanding controversy as to whether such Abs contribute to, or are secondary to, the pathogenesis of RA. To address the potential pathogenicity of human RA associated Abs, we developed a passive transfer model involving mice deficient in the low-affinity inhibitory Fc receptor, FcgammaRIIB. We report that plasma or serum from patients with active RA can induce inflammation and histological lesions in FcgammaRIIB-/- mice consistent with arthritis, and that this pathogenic activity is caused by the immunoglobulin G-rich fraction. Our results suggest that humoral autoimmunity can contribute directly to autoimmune arthritis, and that FcgammaRIIB-/- mice are a promising model to evaluate the arthritogenic potential of human autoAbs. PMID- 16476769 TI - T-independent type II immune responses generate memory B cells. AB - Unlike T-dependent immune responses against protein antigens, T-independent responses against polysaccharides confer long-lasting humoral immunity in the absence of recall responses and are not known to generate memory B cells. Here we report that polysaccharide antigens elicit memory B cells that are phenotypically distinct from those elicited by protein antigens. Furthermore, memory B cell responses against polysaccharides are regulated by antigen-specific immunoglobulin G antibodies. As the generation and regulation of immunologic memory is central to vaccination, our findings help explain the mode of action of the few existing polysaccharide vaccines and provide a rationale for a wider application of polysaccharide-based strategies in vaccination. PMID- 16476771 TI - Strand-biased defect in C/G transversions in hypermutating immunoglobulin genes in Rev1-deficient mice. AB - Somatic hypermutation of Ig genes enables B cells of the germinal center to generate high-affinity immunoglobulin variants. Key intermediates in somatic hypermutation are deoxyuridine lesions, introduced by activation-induced cytidine deaminase. These lesions can be processed further to abasic sites by uracil DNA glycosylase. Mutagenic replication of deoxyuridine, or of its abasic derivative, by translesion synthesis polymerases is hypothesized to underlie somatic hypermutation. Rev1 is a translesion synthesis polymerase that in vitro incorporates uniquely deoxycytidine opposite deoxyuridine and abasic residues. To investigate a role of Rev1 in mammalian somatic hypermutation we have generated mice deficient for Rev1. Although Rev1-/- mice display transient growth retardation, proliferation of Rev1-/- LPS-stimulated B cells is indistinguishable from wild-type cells. In mutated Ig genes from Rev1-/- mice, C to G transversions were virtually absent in the nontranscribed (coding) strand and reduced in the transcribed strand. This defect is associated with an increase of A to T, C to A, and T to C substitutions. These results indicate that Rev1 incorporates deoxycytidine residues, most likely opposite abasic nucleotides, during somatic hypermutation. In addition, loss of Rev1 causes compensatory increase in mutagenesis by other translesion synthesis polymerases. PMID- 16476772 TI - Nuclear actin: to polymerize or not to polymerize. AB - The form and function of actin in the nucleus have been enigmatic for over 30 years. Recently actin has been assigned numerous functional roles in the nucleus, but its form remains a mystery. The intricate relationship between actin form and function in the cytoplasm implies that understanding the structural properties of nuclear actin is elementary to fully understanding its function. In this issue, McDonald et al. (p. 541) use fluorescence recovery after photobleaching (FRAP) to tackle the question of whether nuclear actin exists as monomers or polymers. PMID- 16476773 TI - Drosophila melanogaster gamma-TuRC is dispensable for targeting gamma-tubulin to the centrosome and microtubule nucleation. AB - In metazoans, gamma-tubulin acts within two main complexes, gamma-tubulin small complexes (gamma-TuSCs) and gamma-tubulin ring complexes (gamma-TuRCs). In higher eukaryotes, it is assumed that microtubule nucleation at the centrosome depends on gamma-TuRCs, but the role of gamma-TuRC components remains undefined. For the first time, we analyzed the function of all four gamma-TuRC-specific subunits in Drosophila melanogaster: Dgrip75, Dgrip128, Dgrip163, and Dgp71WD. Grip-motif proteins, but not Dgp71WD, appear to be required for gamma-TuRC assembly. Individual depletion of gamma-TuRC components, in cultured cells and in vivo, induces mitotic delay and abnormal spindles. Surprisingly, gamma-TuSCs are recruited to the centrosomes. These defects are less severe than those resulting from the inhibition of gamma-TuSC components and do not appear critical for viability. Simultaneous cosilencing of all gamma-TuRC proteins leads to stronger phenotypes and partial recruitment of gamma-TuSC. In conclusion, gamma-TuRCs are required for assembly of fully functional spindles, but we suggest that gamma TuSC could be targeted to the centrosomes, which is where basic microtubule assembly activities are maintained. PMID- 16476770 TI - Proteinase-activated receptor 2 modulates neuroinflammation in experimental autoimmune encephalomyelitis and multiple sclerosis. AB - The proteinase-activated receptors (PARs) are widely recognized for their modulatory properties of inflammation and neurodegeneration. We investigated the role of PAR2 in the pathogenesis of multiple sclerosis (MS) in humans and experimental autoimmune encephalomyelitis (EAE) in mice. PAR2 expression was increased on astrocytes and infiltrating macrophages in human MS and murine EAE central nervous system (CNS) white matter (P < 0.05). Macrophages and astrocytes from PAR2 wild-type (WT) and knockout (KO) mice exhibited differential immune gene expression with PAR2 KO macrophages showing significantly higher interleukin 10 production after lipopolysaccharide stimulation (P < 0.001). PAR2 activation in macrophages resulted in the release of soluble oligodendrocyte cytotoxins (P < 0.01). Myelin oligodendrocyte glycoprotein-induced EAE caused more severe inflammatory gene expression in the CNS of PAR2 WT animals (P < 0.05), together with enhanced T cell proliferation and interferon gamma production (P < 0.05), compared with KO littermates. Indeed, PAR2 WT animals showed markedly greater microglial activation and T lymphocyte infiltration accompanied by worsened demyelination and axonal injury in the CNS compared with their PAR2 KO littermates. Enhanced neuropathological changes were associated with a more severe progressive relapsing disease phenotype (P < 0.001) in WT animals. These findings reveal previously unreported pathogenic interactions between CNS PAR2 expression and neuroinflammation with ensuing demyelination and axonal injury. PMID- 16476774 TI - Early aging-associated phenotypes in Bub3/Rae1 haploinsufficient mice. AB - Aging is a highly complex biological process that is believed to involve multiple mechanisms. Mice that have small amounts of the mitotic checkpoint protein BubR1 age much faster than normal mice, but whether other mitotic checkpoint genes function to prevent the early onset of aging is unknown. In this study, we show that several aging-associated phenotypes appear early in mice that are double haploinsufficient for the mitotic checkpoint genes Bub3 and Rae1 but not in mice that are single haploinsufficient for these genes. Mouse embryonic fibroblasts (MEFs) from Bub3/Rae1 haploinsufficient mice undergo premature senescence and accumulate high levels of p19, p53, p21, and p16, whereas MEFs from single haploinsufficient mice do not. Furthermore, although BubR1 hypomorphic mice have less aneuploidy than Bub3/Rae1 haploinsufficient mice, they age much faster. Our findings suggest that early onset of aging-associated phenotypes in mice with mitotic checkpoint gene defects is linked to cellular senescence and activation of the p53 and p16 pathways rather than to aneuploidy. PMID- 16476775 TI - Nucleoplasmic beta-actin exists in a dynamic equilibrium between low-mobility polymeric species and rapidly diffusing populations. AB - Beta-actin, once thought to be an exclusively cytoplasmic protein, is now known to have important functions within the nucleus. Nuclear beta-actin associates with and functions in chromatin remodeling complexes, ribonucleic acid polymerase complexes, and at least some ribonucleoproteins. Proteins involved in regulating actin polymerization are also found in the interphase nucleus. We define the dynamic properties of nuclear actin molecules using fluorescence recovery after photobleaching. Our results indicate that actin and actin-containing complexes are reduced in their mobility through the nucleoplasm diffusing at approximately 0.5 microm2 s(-1). We also observed that approximately 20% of the total nuclear actin pool has properties of polymeric actin that turns over rapidly. This pool could be detected in endogenous nuclear actin by using fluorescent polymeric actin binding proteins and was sensitive to drugs that alter actin polymerization. Our results validate previous reports of polymeric forms of nuclear actin observed in fixed specimens and reveal that these polymeric forms are very dynamic. PMID- 16476776 TI - Mdm38 interacts with ribosomes and is a component of the mitochondrial protein export machinery. AB - Saccharomyces cerevisiae Mdm38 and Ylh47 are homologues of human Letm1, a protein implicated in Wolf-Hirschhorn syndrome. We analyzed the function of Mdm38 and Ylh47 in yeast mitochondria to gain insight into the role of Letm1. We find that mdm38Delta mitochondria have reduced amounts of certain mitochondrially encoded proteins and low levels of complex III and IV and accumulate unassembled Atp6 of complex V of the respiratory chain. Mdm38 is especially required for efficient transport of Atp6 and cytochrome b across the inner membrane, whereas Ylh47 plays a minor role in this process. Both Mdm38 and Ylh47 form stable complexes with mitochondrial ribosomes, similar to what has been reported for Oxa1, a central component of the mitochondrial export machinery. Our results indicate that Mdm38 functions as a component of an Oxa1-independent insertion machinery in the inner membrane and that Mdm38 plays a critical role in the biogenesis of the respiratory chain by coupling ribosome function to protein transport across the inner membrane. PMID- 16476777 TI - A repeated IMP-binding motif controls oskar mRNA translation and anchoring independently of Drosophila melanogaster IMP. AB - Zip code-binding protein 1 (ZBP-1) and its Xenopus laevis homologue, Vg1 RNA and endoplasmic reticulum-associated protein (VERA)/Vg1 RNA-binding protein (RBP), bind repeated motifs in the 3' untranslated regions (UTRs) of localized mRNAs. Although these motifs are required for RNA localization, the necessity of ZBP 1/VERA remains unresolved. We address the role of ZBP-1/VERA through analysis of the Drosophila melanogaster homologue insulin growth factor II mRNA-binding protein (IMP). Using systematic evolution of ligands by exponential enrichment, we identified the IMP-binding element (IBE) UUUAY, a motif that occurs 13 times in the oskar 3'UTR. IMP colocalizes with oskar mRNA at the oocyte posterior, and this depends on the IBEs. Furthermore, mutation of all, or subsets of, the IBEs prevents oskar mRNA translation and anchoring at the posterior. However, oocytes lacking IMP localize and translate oskar mRNA normally, illustrating that one cannot necessarily infer the function of an RBP from mutations in its binding sites. Thus, the translational activation of oskar mRNA must depend on the binding of another factor to the IBEs, and IMP may serve a different purpose, such as masking IBEs in RNAs where they occur by chance. Our findings establish a parallel requirement for IBEs in the regulation of localized maternal mRNAs in D. melanogaster and X. laevis. PMID- 16476778 TI - Huntingtin-HAP40 complex is a novel Rab5 effector that regulates early endosome motility and is up-regulated in Huntington's disease. AB - The molecular mechanisms underlying the targeting of Huntingtin (Htt) to endosomes and its multifaceted role in endocytosis are poorly understood. In this study, we have identified Htt-associated protein 40 (HAP40) as a novel effector of the small guanosine triphosphatase Rab5, a key regulator of endocytosis. HAP40 mediates the recruitment of Htt by Rab5 onto early endosomes. HAP40 overexpression caused a drastic reduction of early endosomal motility through their displacement from microtubules and preferential association with actin filaments. Remarkably, endogenous HAP40 was up-regulated in fibroblasts and brain tissue from human patients affected by Huntington's disease (HD) as well as in STHdhQ(111) striatal cells established from a HD mouse model. These cells consistently displayed altered endosome motility and endocytic activity, which was restored by the ablation of HAP40. In revealing an unexpected link between Rab5, HAP40, and Htt, we uncovered a new mechanism regulating cytoskeleton dependent endosome dynamics and its dysfunction under pathological conditions. PMID- 16476780 TI - A constipation assessment scale for use in pediatric oncology. AB - Constipation is prevalent in pediatric oncology patients because of treatment with vinca alkaloids and/or narcotics and lifestyle changes secondary to disease process. Sequelae of constipation include anorexia, nausea, vomiting, abdominal pain, emergency department visits, and a decrease in quality of life. There are no reliable instruments to measure constipation in children. A pilot study (N = 21) evaluating the presence and severity of constipation and the reliability and validity of a modified version of the adult Constipation Assessment Scale (CAS) in children with cancer was conducted. Patients receiving weekly vinca alkaloids and/or narcotics = 2 times per day were recruited. Initial bowel function assessments included standardized nursing and nutrition assessments, history/physical review, and baseline CAS score repeated at 1 hour to assess test retest reliability. Subsequent assessments included CAS administered 3 times per week and daily patient bowel diaries. Test-retest reliability was evident (r = .93; P = .000). Acceptable construct validity was indicated by a difference in mean CAS scores (t = 4.4, P <.001). Patients reported difficulty with CAS questions and response selections. Symptoms asked on CAS were often not viewed as a problem. PMID- 16476781 TI - Minimizing complications related to fever in the postoperative pediatric oncology patient. AB - Fever is a common postoperative complication that is generally thought to be a benign, self-limiting event. However, for pediatric oncology patients who are often immunocompromised, a postoperative fever may indicate an infection, which can lead to significant complications if not treated promptly. A physical examination, which is an established cost-effective method to rule out infections in the general surgical population, may not be sufficient for oncology patients because clinical symptoms may be atypical or absent in immunocompromised hosts. Although a full septic workup may be unnecessary, additional investigations such as blood cultures may be warranted, and antibiotic therapy should be considered for some or all febrile postoperative cancer patients. No standardized approach to fever management in postoperative oncology patients currently exists, which can present a challenge for those who care for these patients. In the absence of such established practice standards, this article outlines some of the considerations that may be vital in minimizing complications related to fever in the postoperative pediatric oncology patient. PMID- 16476779 TI - The tumor suppressor DAPK inhibits cell motility by blocking the integrin mediated polarity pathway. AB - Death-associated protein kinase (DAPK) is a calmodulin-regulated serine/threonine kinase and possesses apoptotic and tumor-suppressive functions. However, it is unclear whether DAPK elicits apoptosis-independent activity to suppress tumor progression. We show that DAPK inhibits random migration by reducing directional persistence and directed migration by blocking cell polarization. These effects are mainly mediated by an inhibitory role of DAPK in talin head domain association with integrin, thereby suppressing the integrin-Cdc42 polarity pathway. We present evidence indicating that the antimigratory effect of DAPK represents a mechanism through which DAPK suppresses tumors. First, DAPK can block migration and invasion in certain tumor cells that are resistant to DAPK induced apoptosis. Second, using an adenocarcinoma cell line and its highly invasive derivative, we demonstrate DAPK level as a determining factor in tumor invasiveness. Collectively, our study identifies a novel function of DAPK in regulating cell polarity during migration, which may act together with its apoptotic function to suppress tumor progression. PMID- 16476782 TI - Sources of conflict between families and health care professionals. AB - It is essential to examine conflict between patients and health care professionals from the patient's perspective. The purposes of this study were to 1) identify sources of conflict, 2) determine nursing interventions that alleviate conflict, and 3) test a conceptual framework of sources of conflict. This phenomenological study focused on children with cancer and their parents' perceptions of conflicts with health care professionals as well as what they thought helped with such conflicts. Their reports of conflict were compared to C. W. Moore's circle of conflict conceptual framework. A purposive sample of 27 participants (9 children, 14 mothers, and 4 fathers) participated in the study. Study findings showed that conflict occurred between health care professionals and families originating from differences in expectations and desires regarding data, interests, structure, relationships, and values, consistent with C. W. Moore's conceptual framework. Nursing interventions reported by children and parents to be helpful in preventing or alleviating conflict were identified. C. W. Moore's framework may provide a valuable structure for assessing conflict and designing nursing interventions to alleviate conflict. PMID- 16476783 TI - The concept of suffering in children and adolescents with cancer. AB - Survival rates for childhood cancer have increased over the past 2 decades, due in large part to the increase in the intensity and complexity of the treatment modalities used. We can presume that this increase in intensity has produced increased distress or suffering (physical, psychological, social, and spiritual) in the child or adolescent undergoing these therapies for cancer. However, measurement of suffering is more than symptom occurrence, frequency, duration, and severity. An analysis of the concept of suffering is presented. Suffering needs to be defined and measured by self-report (as opposed to parent or staff report) to gain an accurate, complete holistic picture of the nature and scope of the child's and adolescent's suffering. Knowledge of how children and adolescents experience suffering would enable practitioners to design interventions to prevent or ameliorate this suffering. PMID- 16476784 TI - Enteral nutrition and bone marrow transplantation. AB - This retrospective study evaluated the efficacy of enteral nutrition for pediatric patients undergoing the challenging treatment of allogeneic bone marrow transplantation. During the period from January 1999 to May 2000, 15 patients were transplant recipients. On admission to the hospital, 87% of patients were above the 50th percentile for weight for age. Nasogastric tubes were inserted while platelet counts remained greater than 50 x 10(9) mL/L. A specialized elemental formula for pediatric patients was commenced. These feeds were administered continuously and were titrated until caloric requirement or tolerance level had been achieved. During hospitalization for bone marrow transplantation, enteral nutrition was the major form of nutritional support for all patients. Enteral feeds continued even during maximal gut toxicity and were supported with antiemetics and analgesia. There were insignificant weight fluctuations during hospitalization, with 80% of children above the 50th percentile weight for age being discharged. Enteral nutrition via a nasogastric tube was effective in the provision of nutrition during bone marrow transplantation and continues to have an important role in this unit. PMID- 16476785 TI - No nitric oxide for HO-1 from sodium nitroprusside. AB - Nitric oxide (NO) and NO donors were among the first reported inducers of the tissue-protective protein heme oxygenase-1 (HO-1) with a potential for eventual use in humans. Besides other clinically established NO releasing drugs, sodium nitroprusside (SNP) has frequently been employed as an experimental tool to explore effects of NO on HO-1 and other biological targets. In this issue of Molecular Pharmacology, Kim et al. (p. 1633) demonstrate that the effects of SNP on expression of HO-1 are mainly due to free iron released from SNP in aqueous solution, whereas NO plays a negligible role, if any, as the mediator of response to SNP. Downstream effects of iron, after being dissociated from SNP, include increases in intracellular cAMP that are causally linked to subsequent phosphorylation of specific MAPK targets and enhanced HO-1 protein levels. Based on the data reported by Kim et al. (2006), the use of SNP as an experimental tool to mimic intracellular effects of NO should be avoided in the future. This work not only helps revise concepts in NO and HO-1 research but also may direct future efforts to the role of iron and reactive oxygen species in the regulation of adenylyl cyclase. PMID- 16476786 TI - Angiotensin-converting enzyme (ACE) dimerization is the initial step in the ACE inhibitor-induced ACE signaling cascade in endothelial cells. AB - The binding of angiotensin-converting enzyme (ACE) inhibitors to ACE initiates a signaling cascade that involves the phosphorylation of the enzyme on Ser1270 as well as activation of the c-Jun NH2-terminal kinase (JNK) and leads to alterations in gene expression. To clarify how ACE inhibitors activate this pathway, we determined their effect on the ability of the enzyme to dimerize and the role of ACE dimerization in the initiation of the ACE signaling cascade. In endothelial cells, ACE was detected as a monomer as well as a dimer in native gel electrophoresis and dimerization/oligomerization was confirmed using the split ubiquitin assay in yeast. ACE inhibitors elicited a rapid, concentration dependent increase in the dimer/monomer ratio that correlated with that of the ACE inhibitorinduced phosphorylation of ACE. Cell treatment with galactose and glucose to prevent the putative lectin-mediated self-association of ACE or with specific antibodies shielding the N terminus of ACE failed to affect either the basal or the ACE inhibitor-induced dimerization of the enzyme. In ACE-expressing Chinese hamster ovary cells, ACE inhibitors elicited ACE dimerization and phosphorylation as well as the activation of JNK with similar kinetics to those observed in endothelial cells. However, these effects were prevented by the mutation of the essential Zn2+-complexing histidines in the C-terminal active site of the enzyme. Mutation of the N-terminal active site of ACE was without effect. Together, our data suggest that ACE inhibitors can initiate the ACE signaling pathway by inducing ACE dimerization, most probably via the C-terminal active site of the enzyme. PMID- 16476787 TI - Natural resistance, iron and infection: a challenge for clinical medicine. AB - Natural resistance to infection, which does not depend on antibiotics, is a powerful protective mechanism common to all mankind that has been responsible for the survival of our species during countless millennia in the past. The normal functioning of this complex system of phagocytic cells and tissue fluids is entirely dependent on an extremely low level of free ionic iron (10(-18) M) in tissue fluids. This low-iron environment is maintained by the unsaturated iron binding proteins transferrin and lactoferrin, which depend on well-oxygenated tissues, where a relatively high oxidation-reduction potential (Eh) and pH are essential for the binding of ferric iron. Freely available iron is derived from iron overload, free haem compounds, or hypoxia in injured tissue leading to a fall in Eh and pH. This can severely damage or abolish normal bactericidal mechanisms in tissue fluids leading to overwhelming growth of bacteria or fungi. The challenge for clinical medicine is to reduce or eliminate the presence of freely available iron in clinical disease. In injured or hypoxic tissue, treatment with hyperbaric oxygen might prove very useful by increasing tissue oxygenation and restoring normal bactericidal mechanisms in tissue fluids, which would be of huge benefit to the patient. PMID- 16476788 TI - Catalase activity in Candida albicans exposed to antineoplastic drugs. AB - An increased catalase activity in Candida spp. has been suggested as a mechanism that reduces amphotericin B activity. Furthermore, resistance to antifungal agents like amphotericin B has been reported in some cancer patients undergoing chemotherapy treatment. In this study we analysed the influence of chemotherapy agents on catalase activity in Candida albicans, the major species involved in yeast infections. Eight strains of C. albicans isolated from HIV-positive patients were exposed to cyclophosphamide, cytarabine, dacarbazine and methotrexate antineoplastic drugs at the concentrations used during therapy. Catalase activity was measured and compared to the control group. Very significant differences (P < 0.01) were found when C. albicans was exposed to methotrexate (2 microg ml(-1) = 4 microM). For cyclophosphamide (50 microg ml( 1)), cytarabine (1 microg ml(-1)) and dacarbazine (8 microg ml(-1)), no differences were found (P > 0.05) between the control and drug-exposed groups. Although more extensive studies are necessary, these data do suggest that the antineoplastic drug methotrexate contributes to the resistance to antifungal drug therapy by varying catalase activity. PMID- 16476789 TI - Transcriptional profiling of host responses in mouse lungs following aerosol infection with type A Francisella tularensis. AB - Tularaemia caused by inhalation of type A Francisella tularensis bacteria is one of the most aggressive infectious diseases known, but the reasons for the very rapid spread of the organism from the lungs to internal organs and the ensuing mortality are unknown. The present study used the mouse model to examine in detail the host immune response in the lung. After an aerosol challenge with 20 c.f.u. of the type A strain FSC033, all mice developed clinical signs of severe disease, showed weight loss by day 4 of infection and died the next day. Histopathological findings in the lung revealed acute inflammation and intense vasculitis and perivasculitis on day 4. Gene transcriptional changes in the mouse lung samples were examined on days 1, 2 and 4 of infection using a cDNA microarray with 20,600 mouse clones representing 18,500 genes. In total, 424 genes were found to be differentially expressed, some of which were both up- and downregulated at different time points, 192 of which were upregulated and 234 of which were downregulated for at least one time point. A high percentage of selected genes identified by the microarray analysis were confirmed to be differentially regulated by quantitative real-time PCR. Categorization of the differentially expressed genes showed that those preferentially involved in host immune responses were activated extensively on day 4 but hardly or not at all on days 1 and 2. Further analysis revealed that several of the genes upregulated on day 4 are known to depend on gamma interferon or tumour necrosis factor alpha for their regulation. In keeping with this finding, tumour necrosis factor alpha and gamma interferon levels were found to be increased significantly in bronchoalveolar lavage on day 4. PMID- 16476790 TI - Development of a routine laboratory direct detection system of staphylococcal enterotoxin genes. AB - A novel direct detection system has been developed for eight staphylococcal enterotoxin (SE)-encoding genes (sea, seb, sec, sed, see, seg, seh and sei) in milk. Specific detection by real-time PCR was successful for all SE-encoding genes in the reference strains. Furthermore, a novel DNA-preparation method with good reproducibility [coefficients of variation 0.31, 0.99 and 1.21 % at 10(6), 10(4) and 10(2) c.f.u. (ml milk sample)(-1), respectively] was developed to overcome PCR inhibition in the milk samples. The combination of this DNA preparation method and real-time PCR resulted in high sensitivity [between 1.1 x 10(2) and 1.0 x 10(4) c.f.u. (ml milk sample)(-1)] and allowed the completion of the entire procedure within 4 h. Results of an evaluation of this method for the detection of SE-encoding genes using known outbreak milk samples produced results showing good correspondence with the reversed passive latex agglutination assay. In addition, this newly developed system can be applied to clinical samples such as faeces and vomit. Consequently, the system should be useful in the routine direct detection of SE-encoding genes in food-borne-poisoning samples. PMID- 16476792 TI - Evaluation of six agglutination tests for Staphylococcus aureus identification depending upon local prevalence of meticillin-resistant S. aureus (MRSA). AB - Most routine laboratory detection of Staphylococcus aureus isolates is based on rapid agglutination test systems. Failure of agglutination assays to identify meticillin-resistant S. aureus strains (MRSA) has been demonstrated. The aim of this study was to evaluate six commercially available agglutination tests for the detection of meticillin-sensitive S. aureus (MSSA) and mecA-positive MRSA strains. The Dry Spot Staphytect Plus test (Oxoid), the Pastorex Staph Plus test (Bio-Rad), the Slidex Staph-Kit and Slidex Staph Plus test (bioMerieux), the Staphaurex Plus test (Remel) and the Staphylase Test (Oxoid) were used. As determined by pulsed field gel electrophoresis, 52 distinct MRSA strains from five countries, 83 MSSA strains and 150 coagulase-negative staphylococci were included. Species identification and determination of susceptibility patterns were performed using colony morphology, Gram stain, catalase testing, tube coagulase testing, DNase testing, mannitol fermentation, susceptibility testing towards oxacillin by Etest, coagulase gene PCR, fibrinogen receptor gene PCR and PCR of the mecA gene. Sensitivity of the agglutination tests ranged from 82.7 to 100.0 % for MRSA strains and 92.8 to 100.0 % for MSSA strains, respectively. Specificity of the test systems ranged from 91.3 to 99.1 %. None of the six agglutination assays produced correct reactions for all staphylococci tested. Only the Dry Spot Staphytect Plus test correctly identified all 52 MRSA strains. For the other tests kits, sensitivity of MRSA detection was lower than for MSSA isolates. Depending upon the local MRSA prevalence and the parameter of interest (sensitivity or specificity), these test systems may be useful for routine diagnostic purposes. PMID- 16476791 TI - Distribution of Clostridium difficile PCR ribotypes in regions of Hungary. AB - The objective of this survey was to determine the distribution of Clostridium difficile PCR ribotypes present across three Hungarian geographical regions. A total of 105 isolates of C. difficile from diarrhoeal faeces of both inpatients and outpatients were examined. The toxigenic status of the strains was determined by PCR for the tcdA, tcdB, cdtA and cdtB genes in Szeged (Hungary), while strains were subjected to PCR ribotyping in Cardiff (UK). A total of 31 ribotypes were detected among the 105 C. difficile isolates tested. Five PCR ribotypes were distinct from all previously described types, suggesting that they are new. The most common types in Hungary, during the period examined, were PCR ribotype 014 (24.8 %) and PCR ribotype 002 (13.3 %). The distribution of PCR ribotypes differed in the various Hungarian regions: PCR ribotype 012 was frequent (20.7 %) in South Hungary, whereas this type was rare in the Budapest region and was not common to West Hungary. In West Hungary and the Budapest region, PCR ribotype 014 was most frequent (28.9 and 29 %, respectively). PMID- 16476793 TI - Monoclonal antibodies binding to the cell surface of Listeria monocytogenes serotype 4b. AB - Serotype 4b strains of the food-borne pathogen Listeria monocytogenes are responsible for a large portion of sporadic listeric infections and all major food-borne listeriosis outbreaks in humans. Hybridomas were produced from three fusions with lymphocytes of ND4 mice immunized either with the insoluble antigens of L. monocytogenes serotype 4b or with formalin-killed bacterial cells and screened for monoclonal antibodies (mAbs) reactive to L. monocytogenes serotype 4b. A set of 35 mAbs was identified by ELISA as having reactivity with both the insoluble antigen fraction and the whole-cell antigens. Thirteen of these mAbs belonged to immunoglobulin subclass G1 (IgG1), fifteen were IgG2a and seven mAbs were IgM. Only 20 out of the 35 mAbs were capable of detecting protein bands of various sizes ranging from 20 to 88 kDa in Western blots. Two of these mAbs, M2365 and M2367, were capable of binding to cell-surface antigens of live L. monocytogenes serotype 4b, as demonstrated by immunofluorescence microscopy and immunogold transmission electron microscopy. Immunofluorescence microscopy showed that M2365 and M2367 failed to bind to the cell surfaces of Escherichia coli O157 : H7, Salmonella enterica (serotype Typhimurium DT104) or Campylobacter jejuni. Evaluation of the cross-reactions of all 35 mAbs with whole-cell antigens of E. coli O157 : H7, S. Typhimurium, C. jejuni and Listeria innocua by ELISA indicated that the majority of the mAbs, including M2365 and M2367, did not cross-react with E. coli O157 : H7, S. Typhimurium or C. jejuni and showed no or a very weak reaction with L. innocua. Furthermore, M2365 and M2367 showed no reaction with whole-cell antigens derived from L. monocytogenes serotypes 1/2a, 1/2b and 3a, and from Listeria grayi, Listeria ivanovii and Listeria seeligeri, in an ELISA. Collectively, these data suggest that M2365 and M2367 have potential use in the development of immunological methods of laboratory diagnosis for L. monocytogenes serotype 4b in clinical or food samples. PMID- 16476794 TI - Genotyping of Chlamydia trachomatis from clinical specimens in Taiwan. AB - This study was conducted to determine the prevalence and distribution of Chlamydia trachomatis genotypes in Taiwan. Urine and endocervical-swab samples were collected from two hospitals located in northern and southern Taiwan. The genotypes of a total of 145 samples positive for C. trachomatis were analysed by sequencing the omp1 gene and this was successful in 102 samples. Nine different C. trachomatis genotypes were identified. Genotype E was the most prevalent (22 %), followed by D and Da (19 %), F (16 %), J (15 %), K (11 %), G (11 %), H (6 %) and Ba (2 %). There was a geographical difference in the prevalence of genotype H (P < 0.018) between northern and southern Taiwan. Sequence mutation analysis by blast searching against GenBank reference sequences identified 12 genetic variants from a total of 102 omp1 gene sequences. PMID- 16476795 TI - Attachment of Yersinia pestis to human respiratory cell lines is inhibited by certain oligosaccharides. AB - Pneumonic plague is an aggressive disease that is clinically difficult to treat. Inhibition of attachment using oligosaccharide receptor mimics may provide an alternative to antibiotics. The virulent Yersinia pestis strain GB was demonstrated to attach to the murine monocyte cell line (J774A.1) and a range of human respiratory epithelial cell lines: nasal (RPMI-2650), bronchial (BEAS2-B) and alveolar (A549). Attachment was greatest to the A549 and BEAS2-B cell lines. Pre-treatment of the cell lines with tunicamycin reduced attachment by 55-65 %, indicating the importance of cell-surface carbohydrates in adhesion. The cell lines displayed differences in the oligosaccharides that inhibited attachment. p Nitrophenol was the best inhibitor for each cell line. Disaccharides such as GalNAcbeta1-3Gal and GalNAcbeta1-4Gal were also good inhibitors, particularly for the RPMI-2650 cell line. This demonstrates the potential of oligosaccharides as potential anti-adhesion therapeutics. PMID- 16476796 TI - Identification and molecular characterization of mannitol salt positive, coagulase-negative staphylococci from nasal samples of medical personnel and students. AB - The identification of mannitol salt positive, coagulase-negative staphylococci (CNS) is often disregarded when Staphylococcus aureus is screened in clinical samples using mannitol salt agar. However, the emergence of CNS as important human pathogens has indicated that reliable methods for the identification of clinically significant CNS are of great importance in understanding the epidemiology of infections caused by them. The identification and molecular characterization of mannitol salt positive CNS from nasal samples of medical personnel and students is reported here. A total of 84 mannitol salt positive staphylococcal isolates were obtained from 240 nasal samples, of which 15 were CNS. The API STAPH system classified the CNS isolates into six species, and one third of the isolates were identified with confidence levels of <80 %. 16S-23S rRNA intergenic spacer length polymorphism analysis (ITS-PCR) identified only two species (Staphylococcus haemolyticus and Staphylococcus saprophyticus). This identification was confirmed by antibiotyping, species-specific PCR and PFGE. The results from this study indicate that ITS-PCR is a potentially useful and reliable tool, enabling hospital laboratories to obtain rapid, full and accurate identification of CNS at the species level. PMID- 16476797 TI - Association between cell-bound haemolysin and cell-detaching activity of Escherichia coli isolated from children. AB - Haemolysin alpha (HlyA) produced by cell-detaching Escherichia coli, a putative new class of enteric pathogen, is considered to be the main factor responsible for detachment of cells cultured in vitro. HlyA is one of the few E. coli proteins actively secreted into the medium during exponential growth. In the present study 27 HlyA-positive E. coli isolates, randomly selected from stool specimens, produced a cell-bound haemolysin that was detectable during the exponential and stationary growth phases. The influence of both cell-free and cell-bound haemolysins of the selected isolates on cell-detaching activity of E. coli in vitro was determined. The results suggest that cell-bound haemolysin rather than cell-free HlyA was responsible for the cell-detaching activity of E. coli strains tested. PMID- 16476798 TI - Random amplified polymorphic DNA analysis of Campylobacter jejuni and Campylobacter coli isolated from healthy cattle and sheep. AB - The genetic heterogeneity among Campylobacter jejuni and Campylobacter coli isolates obtained from apparently healthy cattle and sheep was investigated by random amplified polymorphic DNA (RAPD) analysis. A total of 348 Campylobacter isolates, consisting of C. jejuni (n = 218) and C. coli (n = 130), were analysed. All these isolates were successfully typed by RAPD analysis. The total numbers of band patterns defined by RAPD in cattle and sheep were 42 and 45, respectively. Of the 42 distinct types obtained from cattle, 37 types were observed in C. jejuni isolates (n = 115), and the remaining 5 were in C. coli isolates (n = 30). Of 45 distinct types obtained from sheep, 21 types were observed in C. jejuni isolates (n = 103), and 24 were in C. coli isolates (n = 100). It was concluded that a high degree of heterogeneity existed among the C. jejuni and C. coli isolates of healthy cattle and sheep. PMID- 16476799 TI - Interstitial pulmonary inflammation due to Microbacterium sp. after heart transplantation. AB - A coryneform bacterium was isolated from the bronchoalveolar aspirate of a patient with interstitial pulmonary inflammation. Commercial systems identified the isolate as Corynebacterium sp. or Aureobacterium sp./Corynebacterium aquaticum, but 16S rRNA gene analysis unequivocally attributed it to the genus Microbacterium. This represents the first documented case of Microbacterium pulmonary infection. PMID- 16476800 TI - Severe case of Mediterranean spotted fever in Greece with predominantly neurological features. AB - Rickettsia conorii is endemic in Greece, though only a few cases of infection have been published to date. The case of a 58-year-old man from northern Greece with a severe form of Mediterranean spotted fever and rapid neurological deterioration is presented here. The patient received antibiotic treatment with doxycycline, showing immediate clinical and laboratory improvement. Diagnosis was confirmed later, during the second week after disease onset, by detection of elevated titres of IgM and IgG antibodies against R. conorii using an indirect immunofluorescence assay. PMID- 16476801 TI - Cerebral mass in a 13-year-old girl following long-term sojourn in the Tropics. AB - Cysticercosis of the central nervous system is the main cause of late-onset epilepsy in tropical countries. The case of a 13-year-old German girl with a generalized seizure following long-term sojourns in the Tropics is reported. Cranial imaging showed two cerebral lesions with central calcifications. Serological, molecular and cultural examination of cerebrospinal fluid and blood was negative for various parasites, fungi and bacteria including mycobacteria. Histopathological examination after neurosurgical resection revealed calcareous bodies pathognomonic for platyhelminths, in particular tapeworms. Taken together, the radiological and histopathological findings indicate infection with cysticerci, the larvae of Taenia solium. PMID- 16476802 TI - Lack of association between presentation of diarrhoeal symptoms and faecal isolation of Aeromonas spp. amongst outpatients in Hong Kong. PMID- 16476803 TI - Imipenem-resistant Pseudomonas aeruginosa producing IMP-1 metallo-beta-lactamases and lacking the outer-membrane protein OprD. PMID- 16476805 TI - What's in a NAIM? Hashimoto encephalopathy, steroid-responsive encephalopathy associated with autoimmune thyroiditis, or nonvasculitic autoimmune meningoencephalitis? PMID- 16476804 TI - Pyridostigmine treatment trial in neurogenic orthostatic hypotension. AB - BACKGROUND: Midodrine hydrochloride is the only drug demonstrated in a placebo controlled treatment trial to improve orthostatic hypotension (OH) but it significantly worsens supine hypertension. By enhancing ganglionic transmission, pyridostigmine bromide can potentially ameliorate OH without worsening supine hypertension. OBJECTIVE: To evaluate the efficacy of a single 60-mg dose of pyridostigmine bromide, alone or in combination with a subthreshold (2.5 mg) or suprathreshold (5 mg) dose of midodrine hydrochloride, compared with placebo. DESIGN: We report a double-blind, randomized, 4-way cross-over study of pyridostigmine in the treatment of neurogenic OH. A total of 58 patients with neurogenic OH were enrolled. After 1 day of baseline measurements, patients were given 4 treatments (3 active treatments [60 mg of pyridostigmine bromide; 60 mg of pyridostigmine bromide and 2.5 mg of midodrine hydrochloride; 60 mg of pyridostigmine bromide and 5 mg of midodrine hydrochloride] and a placebo) in random order on successive days. Blood pressure (BP) and heart rate were measured, both supine and standing, immediately before treatment and hourly for 6 hours after the treatment was given. RESULTS: No significant differences were seen in the supine BP, either systolic (P = .36) or diastolic (P = .85). In contrast, the primary end point of the fall in standing diastolic BP was significantly reduced (P = .02) with treatment. Pairwise comparison showed significant reduction by pyridostigmine alone (BP fall of 27.6 mm Hg vs 34.0 mm Hg with placebo; P = .04) and pyridostigmine and 5 mg of midodrine hydrochloride (BP fall of 27.2 mm Hg vs 34.0 mm Hg with placebo; P = .002). Standing BP improvement significantly regressed with improvement in OH symptoms. CONCLUSIONS: Pyridostigmine significantly improves standing BP in patients with OH without worsening supine hypertension. The greatest effect is on diastolic BP, suggesting that the improvement is due to increased total peripheral resistance. PMID- 16476806 TI - Apolipoprotein E and dementia in Parkinson disease: a meta-analysis. AB - OBJECTIVE: To understand the relationship of apolipoprotein E (APOE) polymorphism to dementia in Parkinson disease (PD) because the APOE epsilon4 allele is linked to Alzheimer disease. DATA SOURCE: We reviewed MEDLINE, BIOSIS Previews, and ISI Web of Science from January 1, 1966, to May 7, 2004, supplemented by citation analysis from retrieved articles. STUDY SELECTION: Case-control studies using clinical or pathologic criteria for PD and dementia, and with complete APOE genotype frequencies data. DATA EXTRACTION: We compared estimated prevalence odds ratios for dementia in PD in relation to each allele. We also looked for evidence of heterogeneity and publication bias and performed a stratified analysis on several study characteristics. DATA SYNTHESIS: Data analyses suggest publication bias and heterogeneity of source data for the epsilon4 allele (homogeneity P = .2; Begg and Mazumdar, P = .06; and Egger et al, P = .1). The estimated odds ratios for development of dementia in PD are 1.6 for epsilon4 (95% confidence interval, 1.0-2.5); 1.3 for epsilon2 (95% confidence interval, 0.73-2.4); and 0.54 for epsilon3 (95% confidence interval, 0.18-1.6). The odds ratio estimates for epsilon4 were higher for studies published in 1996 or later (2.3 vs 1.0) and for studies conducted outside North American sites (2.4 vs 1.2). CONCLUSIONS: The APOE epsilon4 allele appears to be associated with a higher prevalence of dementia in PD. Publication bias and heterogeneous source data may, however, confound this conclusion. Confirmatory studies that use standardized and validated diagnostic criteria for dementia in PD are needed. PMID- 16476807 TI - Steroid-responsive encephalopathy associated with autoimmune thyroiditis. AB - BACKGROUND: Steroid-responsive encephalopathy associated with autoimmune thyroiditis (SREAT), often termed Hashimoto encephalopathy, is a poorly understood and often misdiagnosed entity. OBJECTIVE: To characterize the clinical, laboratory, and radiologic findings in patients with SREAT to potentially improve recognition of this treatable entity. DESIGN: Retrospective analysis of clinical features and diagnostic test data. SETTING: Two affiliated tertiary care referral institutions. PATIENTS: Twenty consecutive (6 male) patients diagnosed as having SREAT from 1995 to 2003. MAIN OUTCOME MEASURES: Clinical features and ancillary test findings associated with SREAT. RESULTS: The median age at disease onset was 56 years (range, 27-84 years). The most frequent clinical features were tremor in 16 (80%), transient aphasia in 16 (80%), myoclonus in 13 (65%), gait ataxia in 13 (65%), seizures in 12 (60%), and sleep abnormalities in 11 (55%). All patients were assigned an alternative misdiagnosis at presentation, most commonly viral encephalitis (n = 5), Creutzfeldt-Jakob disease (n = 3), or a degenerative dementia (n = 4). The most frequent laboratory abnormalities were increased liver enzyme levels in 11, increased serum sensitive thyroid-stimulating hormone levels in 11, and increased erythrocyte sedimentation rate in 5. In only 5 patients (25%) did cerebrospinal fluid abnormalities suggest an inflammatory process. Magnetic resonance imaging abnormalities believed to be related to the encephalopathy were present in 5 patients (26%). CONCLUSIONS: The clinical, laboratory, and radiologic findings associated with SREAT are more varied than previously reported. Misdiagnosis at presentation is common. This treatable syndrome should be considered even if the serum sensitive thyroid stimulating hormone level and erythrocyte sedimentation rate are normal, the cerebrospinal fluid profile does not suggest an inflammatory process, and neuroimaging results are normal. Until the pathophysiologic mechanism of this and other autoimmune encephalopathies is better characterized, we believe that descriptive terms that reflect an association rather than causation are most appropriate for this syndrome. PMID- 16476808 TI - Levodopa-associated dyskinesia risk among Parkinson disease patients in Olmsted County, Minnesota, 1976-1990. AB - BACKGROUND: The threat of levodopa-induced dyskinesias often influences early treatment decisions in those with Parkinson disease. OBJECTIVE: To determine the long-term risks of levodopa-associated dyskinesias of any severity, dyskinesias sufficient to require medication adjustment, and dyskinesias failing medication adjustments. DESIGN: The medical records linkage system of the Rochester Epidemiology Project was used to identify all incident Parkinson disease patients treated with levodopa (1976-1990). All records were independently reviewed by 2 neurologists who recorded demographic and drug data, dates when dyskinesias were initially identified, and dates when dyskinesias were sufficient to require medication changes; dyskinesias not controlled by drug adjustments were also tabulated. RESULTS: We identified 126 incident Parkinson disease patients treated with levodopa for at least 2 months. By Kaplan-Meier analysis, the estimated rate of dyskinesias was 30% by 5 treatment years and 59% by 10 years. However, the rate of dyskinesias requiring medication adjustment was estimated to be only 17% by 5 years and 43% by 10 years. At 10 treatment years, the rate of dyskinesias that could not be controlled with medication adjustments was estimated at only 12%. An increased risk was associated with younger age and higher initial levodopa dose, but not with sex. CONCLUSIONS: Levodopa-associated dyskinesias can be expected to develop in nearly 60% of patients in our community after 10 years, but these will be severe enough to require medication adjustments in only 43% of patients. At 10 treatment years, nearly 90% of these patients can expect to be spared dyskinesias that could not be controlled by drug adjustments. This population-based study suggests dyskinesia risk may not be a major concern for most Parkinson disease patients. PMID- 16476809 TI - A randomized controlled trial of etilevodopa in patients with Parkinson disease who have motor fluctuations. AB - BACKGROUND: Motor fluctuations are a common complication in patients with Parkinson disease (PD) receiving long-term levodopa therapy. Slowed gastric emptying and poor solubility of levodopa in the gastrointestinal tract may delay the onset of drug benefit after dosing. Etilevodopa is an ethyl-ester prodrug of levodopa that has greater gastric solubility, passes quickly into the small intestine, is rapidly hydrolyzed to levodopa, and has a shortened time to maximum levodopa concentration. OBJECTIVE: To determine the efficacy, safety, and tolerability of etilevodopa in patients with PD who have motor fluctuations. DESIGN: A double-blind, randomized, comparative clinical trial. SETTING: Forty four sites in the United States and Canada. PATIENTS: Three hundred twenty-seven patients with PD who had a latency of at least 90 minutes total daily time to "on" (TTON) after levodopa dosing. INTERVENTION: Treatment with either etilevodopa-carbidopa or levodopa-carbidopa for 18 weeks. MAIN OUTCOME MEASURE: Change from baseline in total daily TTON as measured using home diaries. RESULTS: The reduction in mean total daily TTON from baseline to treatment was 0.58 hour in the etilevodopa-carbidopa group and 0.79 hour in the levodopa-carbidopa group (P = .24). There was no significant difference between the etilevodopa-carbidopa and levodopa-carbidopa groups in the reduction of response failures (-6.82% vs 4.69%; P = .20). Total daily "off" time improved in the etilevodopa-carbidopa ( 0.85 hour) and levodopa-carbidopa (-0.87 hour) groups without an increase in on time with troublesome dyskinesias. CONCLUSION: Despite the theoretical pharmacokinetic advantage of etilevodopa, there was no improvement in TTON, response failures, or off time compared with levodopa. PMID- 16476810 TI - Incidence and causes of nondegenerative nonvascular dementia: a population-based study. AB - BACKGROUND: Information on the incidence of nondegenerative and nonvascular dementia is limited. DESIGN: We used the records-linkage system of the Rochester Epidemiology Project to ascertain incident cases of dementia in Rochester, Minn, from January 1, 1990, through December 31, 1994. To define causes of dementia, we reviewed all diagnoses, imaging study results, laboratory test results, and clinical courses, as recorded historically in the patient dossier. RESULTS: We found 560 incident cases of dementia, and 60 of them (10.7%) had onset before the age of 70 years (younger-onset group). Forty-three cases (7.7%) were due to nondegenerative nonvascular causes and represented 30.0% of the total in the younger-onset group, but only 5.0% of the total in the older-onset group (aged 70 99 years). The most common nondegenerative nonvascular causes were cancer with or without brain metastases (n = 13), chronic alcoholism (n = 7), and chronic mental illness (n = 11). There were no cases of dementia due to normal-pressure hydrocephalus, subdural hematoma, hypothyroidism, vitamin B12 deficiency, or neurosyphilis. There were 2 individuals with acute confusion due to subdural hematoma and 1 with hypothyroidism whose cognition normalized with therapy. CONCLUSIONS: Nondegenerative nonvascular causes were more common than expected in patients with a younger onset of dementia. None of the patients with dementia reverted to normal with treatment of the putative reversible cause. PMID- 16476811 TI - Relation of quantitative indexes of concurrent alpha-synuclein abnormalities to clinical outcome in autopsy-proven Alzheimer disease. AB - BACKGROUND: Lewy bodies (LBs) and Lewy neurites are frequent concomitant neuropathologic observations in clinical and neuropathologically defined Alzheimer disease (AD), but their relation to clinical features in AD is uncertain. Most studies used semiquantitative measures to determine the presence or absence of LB abnormalities. OBJECTIVE: To determine the clinical consequences of LB abnormalities in the setting of AD. DESIGN: Prospective study. SETTING: Three outpatient research and treatment centers. PARTICIPANTS: Fourteen autopsy cases with a pathologic diagnosis of AD abnormalities and concomitant LBs followed semiannually for up to 8 years (mean age at intake, 72 years; mean age at death, 77 years; mean education, 15 years; 12 women). MAIN OUTCOME MEASURES: The modified Mini-Mental State Examination was used to assess cognitive function. The Unified Parkinson Disease Rating Scale was used to rate extrapyramidal motor signs. Hallucinations were evaluated using the Columbia University Scale for Psychopathology in Alzheimer's Disease. Time from the first evaluation in which diagnostic criteria for probable AD were met to death was used to determine illness duration. Quantitative measures of LB abnormalities were obtained for the frontal cortex, entorhinal cortex, substantia nigra, and hippocampus. RESULTS: Independent-samples t tests were used to assess whether the degree of LB abnormality varied as a function of the presence or absence of hallucinations and extrapyramidal signs. Pearson r correlations were run to examine whether there was a relation among LB abnormalities, cognitive function, and illness duration. There was no relation between quantitative neuropathologic indexes of LB abnormalities and clinical outcome. CONCLUSION: The variability of clinical features in AD was not related to the presence or degree of LB abnormalities. PMID- 16476812 TI - Vertebral artery compression of the medulla. AB - BACKGROUND: Intracranial arteries in the subarachnoid space may compress the brain parenchyma and cranial nerves. Most arterial compressive lesions have been attributed to dolichoectasia in the vertebral-basilar system, and prior reports have concentrated on the pressure effects of basilar artery ectasia. Much less is known about vertebral artery compression of the medulla. OBJECTIVE: To describe a series of patients with vertebral arteries compressing the medulla oblongata. DESIGN: Prospective case studies. SETTING: Tertiary care center. PATIENTS: Nine symptomatic patients, 4 men and 5 women, between the ages of 32 and 79 years. MAIN OUTCOME MEASURES: Clinical phenomena, radiographic findings, treatment, and outcomes. RESULTS: We found that compression most commonly occurs at the ventrolateral surface. The clinical features can be transient or permanent and are predominantly motor and cerebellar or vestibular, but a poor correlation exists between the clinical findings and the severity or extent of impingement. The vertebral arteries were angulated, tortuous, or dilated but not necessarily dolichoectatic to cause obvious indentation. Seven patients were treated with antiplatelets and anticoagulants or analgesics, whereas 2 underwent microvascular decompression, resulting in temporary or no relief. One surgical patient developed cranial nerve complications. Among the medically treated patients, none had progression of deficits, and those with single episodes had no recurrence of symptoms. CONCLUSION: This study is the largest collection, to our knowledge, of patients with medullary vascular compression. Further studies are needed to estimate its frequency, natural course, and preferred management. PMID- 16476813 TI - Association of white matter hyperintensity volume with decreased cognitive functioning: the Framingham Heart Study. AB - OBJECTIVE: To examine the relationship between white matter hyperintensity (WMH) volume on magnetic resonance images and cognitive tests in a large, population based sample. METHODS: Quantitative magnetic resonance imaging and neuropsychological evaluations were performed in 1820 dementia- and stroke-free participants from the Framingham Offspring Cohort. The WMH volume relative to total cranial volume was computed; WMH volumes more than 1 SD above the age predicted mean were defined as large. Adjusting for age, sex, education, height, and Framingham Stroke Risk Profile, we examined the relationship between WMH and 3 cognitive factors derived from a neuropsychological test battery (verbal memory, visuospatial memory and organization, and visual scanning and motor speed) and 3 individual measures of new learning, abstract reasoning, and naming. RESULTS: Compared with those with no or little WMH volume, participants with large WMH volume performed worse on the cognitive factors of visuospatial memory and organization (P = .04) and visual scanning and motor speed (P = .01), as well as on new learning (P = .04), but not on verbal memory (P = .52). CONCLUSIONS: In this younger community-based population of nondemented individuals, those with large WMH volume, as compared with those with less or no WMH volumes, performed significantly worse in cognitive domains generally associated with frontal lobe systems and, to a lesser extent, the medial temporal area. Further study will clarify whether large WMH volume and associated cognitive impairment lead to future risk of stroke or dementia. PMID- 16476814 TI - Spectrum of brain changes in patients with congenital muscular dystrophy and FKRP gene mutations. AB - OBJECTIVES: To report the spectrum of brain magnetic resonance imaging findings in 13 patients with congenital muscular dystrophy and FKRP gene mutations and to explore possible genotype-phenotype correlations. DESIGN: We retrospectively reviewed brain magnetic resonance imaging in patients with congenital muscular dystrophy and FKRP gene mutations. PATIENTS: Thirteen patients with congenital muscular dystrophy and mutations in the FKRP gene. RESULTS: Five of the 13 patients had the typical phenotype originally described for congenital muscular dystrophy (MDC1C) with normal intelligence and normal brain magnetic resonance imaging while 3 other patients had isolated cerebellar cysts and mental retardation without any other sign of posterior fossa of supratentorial abnormalities. In the remaining 5 patients cerebellar cysts were associated with structural brain changes involving the posterior fossa and the cortex, ranging from focal unilateral periventricular nodular heterotopia to marked cerebellar dysplasia and pontine hypoplasia. In 2 of these 5 patients the severity and distribution of changes resembled muscle-eye-brain disease in 1 patient who had mild Walker-Warburg syndrome. The distribution of FKRP gene mutations identified in this group of patients did not reveal any obvious association with the severity of central nervous system involvement. CONCLUSIONS: The severity of central nervous system involvement observed in our patients in contrast broadly reflected the severity of the disruption of alpha-dystroglycan glycosylation. In particular, dystroglycan expression was almost absent in the patients with muscle eye-brain diseaselike phenotype and less severely reduced in the patients with congenital muscular dystrophy (MDC1C) with or without cerebellar cysts. This study further highlights the central role that dystroglycan has in neuronal migration. PMID- 16476815 TI - The G93C mutation in superoxide dismutase 1: clinicopathologic phenotype and prognosis. AB - BACKGROUND: Twenty percent of familial amyotrophic lateral sclerosis (ALS) is caused by mutations in the superoxide dismutase 1 gene (SOD1). Few data exist on their clinicopathologic phenotypes. OBJECTIVES: To determine the clinical and pathologic phenotype associated with the G93C mutation in SOD1 and to compare survival in familial ALS related to this mutation with survival in other ALS subgroups. DESIGN: Retrospective study. SETTING: Tertiary referral center for neuromuscular disorders. PATIENTS: Twenty patients with the G93C mutation for whom clinical data were available and 1 patient with pathologic data. MAIN OUTCOME MEASURES: Characteristics and survival compared with other ALS subgroups, adjusting for known prognostic factors. RESULTS: The G93C mutation was associated with a purely lower motor neuron phenotype without bulbar involvement. Presence of the mutation independently predicted longer survival compared with other ALS subgroups. Pathologic examination showed degeneration of the anterior horn, spinocerebellar tracts, and posterior funiculi, with minimal involvement of corticospinal tracts and no degeneration of brainstem motor nuclei. Survival motor neuron gene copy number had no significant influence on age at onset or survival in patients with the G93C mutation. CONCLUSIONS: These findings add to the knowledge of SOD1-related familial ALS and demonstrate further clinicopathologic variability between different SOD1 mutations. Finally, they demonstrate the independent prognostic value of the G93C mutation. PMID- 16476816 TI - Absence of rapid eye movement sleep behavior disorder in 11 members of the pallidopontonigral degeneration kindred. AB - BACKGROUND: Rapid eye movement sleep behavior disorder (RBD) is a parasomnia that is manifested by dream enactment behavior. The electrophysiologic substrate for RBD on polysomnography is rapid eye movement sleep without atonia. Rapid eye movement sleep behavior disorder likely stems from neuronal network dysfunction in the brainstem, although it is not yet clear which specific networks are involved. Rapid eye movement sleep behavior disorder is often associated with the sporadic synucleinopathies but rarely associated with the sporadic tauopathies. There are no reports on the possible association of rapid eye movement sleep without atonia and RBD with any familial tauopathy. OBJECTIVE: To characterize the clinical sleep and polysomnography features in a kindred with a familial tauopathy. METHODS: We performed standard polysomnography in 11 members of the pallidopontonigral degeneration kindred irrespective of any sleep-related complaints. Neuropathologic findings were analyzed in those who subsequently underwent autopsy. RESULTS: Six affected and 5 genealogically at-risk family members were studied. None of the 11 had a history of dream enactment behavior. Nine of the 11 members attained sufficient rapid eye movement sleep on polysomnography, and the electrophysiologic features of rapid eye movement sleep without atonia and behavioral manifestations of RBD were absent in all subjects. Neuropathologic examination of 4 affected individuals revealed marked nigral degeneration in 3 along with mild degenerative changes in the locus coeruleus, pontine nuclei and tegmentum, and medullary tegmentum. CONCLUSIONS: These findings argue against nigral degeneration being the primary cause of RBD. The absence of the historical, electrophysiologic, and behavioral manifestations of RBD in this kindred provides further evidence that RBD is rare in the sporadic and familial tauopathies. The difference in frequencies of RBD associated with the synucleinopathies compared with the tauopathies suggests differences in the selective vulnerability of brainstem circuits between the synucleinopathies and tauopathies. PMID- 16476817 TI - Heterogeneous phenotype in a family with compound heterozygous parkin gene mutations. AB - BACKGROUND: Mutations in the parkin gene (PRKN) cause autosomal recessive early onset Parkinson disease (EOPD). OBJECTIVE: To investigate the presence of mutations in the PRKN gene in a white family with EOPD and the genotype-phenotype correlations. DESIGN: Twenty members belonging to 3 generations of the EOPD family with 4 affected subjects underwent genetic analysis. Direct genomic DNA sequencing, semiquantitative polymerase chain reaction, real-time quantitative polymerase chain reaction, and reverse-transcriptase polymerase chain reaction analyses were performed to identify the PRKN mutation. RESULTS: Compound heterozygous mutations (T240M and EX 5_6 del) in the PRKN gene were identified in 4 patients with early onset (at ages 30-38 years). Although heterozygous T240M and homozygous EX 5_6 del mutations in the PRKN gene have been previously described, this is, to our knowledge, the first report of these mutations in compound heterozygotes. The phenotype of patients was that of classic autosomal recessive EOPD characterized by beneficial response to levodopa, relatively slow progression, and motor complications. All heterozygous mutation carriers (T240M or EX 5_6 del) and a 56-year-old woman who was a compound heterozygous mutation carrier (T240M and EX 5_6 del) were free of any neurological symptoms. CONCLUSIONS: Compound heterozygous mutations (T240M and EX 5_6 del) in the PRKN gene were found to cause autosomal recessive EOPD in 4 members of a large white family. One additional member with the same mutation, who is more than 10 years older than the mean age at onset of the 4 affected individuals, had no clinical manifestation of the disease. This incomplete penetrance has implications for genetic counseling, and it suggests that complex gene-environment interactions may play a role in the pathogenesis of PRKN EOPD. PMID- 16476819 TI - Aortic rupture in mitochondrial encephalopathy, lactic acidosis, and stroke-like episodes. AB - BACKGROUND: Microangiopathy has been well described in the brain and muscle of patients with mitochondrial encephalopathy, lactic acidosis, and stroke-like episodes (MELAS). OBJECTIVE: To describe a patient with the common A3243G/MELAS point mutation who had aortic rupture and whose mother also died of large vessel rupture. DESIGN: Case report. SETTING: Collaboration between a primary care hospital and 2 academic tertiary care hospitals. RESULTS: Histologically, there was marked disarray of the smooth muscle architecture of the aorta, and immunohistochemical staining with antibodies against the mitochondrial DNA encoded cytochrome-C oxidase I subunit showed uniformly decreased immunostaining of the endothelial and smooth muscle cells of the aorta and vasa vasorum. Polymerase chain reaction and restriction fragment length polymorphism analysis showed that the mutation load was 40.5% in blood but 85.3% in the blood vessels. CONCLUSIONS: The severe vasculopathy in this patient is probably directly related to the high mutation load in the blood vessels. Although aortic rupture is an unusual manifestation of MELAS, it is an important potential complication in patients undergoing minor surgical procedures. PMID- 16476818 TI - Phenotypic variability among adult siblings with Sjogren-Larsson syndrome. AB - BACKGROUND: Sjogren-Larsson syndrome (SLS) is an early childhood-onset disorder with ichthyosis, mental retardation, spastic paraparesis, macular dystrophy, and leukoencephalopathy caused by the deficiency of fatty aldehyde dehydrogenase due to mutations in the ALDH3A2 gene (the gene that encodes microsomal fatty aldehyde dehydrogenase). Cerebral proton magnetic resonance spectroscopy in those with SLS demonstrates an abnormal white matter peak at 1.3 ppm, consistent with long-chain fatty alcohol accumulation. OBJECTIVE: To define the clinical course and proton magnetic resonance spectroscopic findings of SLS in adults. DESIGN AND SETTING: Case series in a tertiary care center. PATIENTS: Six siblings of a consanguineous Arab family with early childhood-onset SLS who carry the 682C-->T mutation in the ALDH3A2 gene were reinvestigated in adulthood. RESULTS: The 6 affected siblings ranged in age from 16 to 36 years. All exhibited the typical clinical and imaging manifestations of SLS, but their severity markedly varied. Neurological involvement was apparently nonprogressive, and its severity showed no correlation with age. Cerebral proton magnetic resonance spectroscopy showed a lipid peak at 1.3 ppm, with decreasing intensity in the older siblings. CONCLUSION: These observations document significant clinical variability and the nonprogressive neurological course of SLS in adult siblings with the same ALDH3A2 genotype, and demonstrate possible correlation of proton magnetic resonance spectroscopic changes with age, suggesting unknown pathogenic mechanisms to compensate for the responsible biochemical defect in this disease. PMID- 16476820 TI - A missense mutation in the coiled-coil domain of the KIF5A gene and late-onset hereditary spastic paraplegia. AB - BACKGROUND: To our knowledge, up to now, only 2 mutations in the KIF5A gene, a member of the kinesin superfamily, have been identified as the molecular cause of early-onset autosomal dominant hereditary spastic paraparesis (ADHSP). OBJECTIVE: To assess the genetic defect in a family with late-onset ADHSP. PATIENTS AND METHODS: Only the proband agreed to undergo complete neurological testing and mutational analysis. The proband was screened for mutations in the spastin, atlastin, NIPA1, and KIF5A genes, either by denaturing high-performance liquid chromatography or sequence analysis. RESULTS: The history of the family was consistent with ADHSP characterized by late onset of the disease. Mutational analysis results were negative for the spastin, atlastin, and NIPA1 genes but identified a missense mutation (c.1082C>T) in the coiled-coil coding region of the KIF5A gene. CONCLUSIONS: This finding enlarges the phenotypic spectrum of ADHSP linked to KIF5A and enhances the role of that gene in the epidemiology of this disease. We propose that the KIF5A gene should be routinely analyzed in patients with hereditary spastic paraplegia negative for spastin and atlastin mutations. PMID- 16476821 TI - Human growth hormone-related iatrogenic Creutzfeldt-Jakob disease with abnormal imaging. AB - BACKGROUND: Although more than 160 cases of iatrogenic Creutzfeldt-Jakob disease (iCJD) from human growth hormone (hGH) treatment have been documented, to our knowledge abnormal cerebellar findings on magnetic resonance imaging (MRI) have not been reported. OBJECTIVE: To report a case of hGH-related iCJD with abnormal cerebellar MRI findings on fluid-attenuated inversion recovery (FLAIR) and diffusion-weighted MRI (DWI). DESIGN: Case report. SETTING: Outpatient neurology clinic at a university medical center. PATIENT: A 33-year old man who had subacute gait ataxia and blurred vision. RESULTS: Beginning 19 years prior, this patient had received cadaveric pituitary-derived hGH treatment for at least 5 years. Magnetic resonance imaging revealed FLAIR and DWI abnormalities, particularly in the cerebellum. He died 7 months after disease onset of autopsy confirmed iCJD. Pathological changes corresponded largely to MRI findings. CONCLUSIONS: To our knowledge, this is the first case of hGH-related iCJD with FLAIR and DWI abnormalities within the cerebellum. As symptoms referable to the cerebellum occur early in iCJD, it suggests that these MRI sequences may allow earlier diagnosis of this form of prion disease. PMID- 16476822 TI - Hyperostosis frontalis interna. PMID- 16476823 TI - The "eye-of-the-tiger" sign is not pathognomonic of the PANK2 mutation. PMID- 16476824 TI - Eagle syndrome: an unusual cause of head and neck pain. PMID- 16476825 TI - An early description of painful neuropathy in Hittite tablets. PMID- 16476826 TI - Impulse control disorders and dopaminergic drugs. PMID- 16476827 TI - Gambling and Parkinson disease. PMID- 16476828 TI - Association between pathologic gambling and parkinsonian therapy as detected in the Food and Drug Administration Adverse Event database. PMID- 16476829 TI - Rebound inflammation and the risk of ischemic stroke after discontinuation of aspirin therapy. PMID- 16476830 TI - Do oncologists believe new cancer drugs offer good value? AB - BACKGROUND: Substantial debate centers on the high cost and relative value of new cancer therapies. Oncologists play a pivotal role in treatment decisions, yet it is unclear whether they perceive high-cost new treatments to offer good value or how therapeutic costs factor into their treatment recommendations. METHODS: We surveyed 139 academic medical oncologists at two academic hospitals in Boston. We asked respondents to provide estimates for the cost and effectiveness of bevacizumab and whether they believed the treatment offered "good value." We also asked respondents to judge how large a gain in life expectancy would justify a hypothetical cancer drug that costs $70,000 a year. Using this information, we calculated implied cost-effectiveness thresholds. Finally, we explored respondents' views on the role of cost in treatment decisions. RESULTS: Ninety academic oncologists (65%) completed the survey. Seventy-eight percent stated that patients should have access to "effective" care regardless of cost. Implied cost-effectiveness thresholds, derived from the bevacizumab and hypothetical scenarios, averaged roughly $300,000 per quality-adjusted-life-year (QALY). Only 25% of oncologists felt that bevacizumab offered "good value." CONCLUSIONS: A majority of academic oncologists stated that cost does not influence their clinical practice, nor should it limit access to "effective" care. Yet respondents did not consider all effective drugs to be of good value. Implied cost-effectiveness thresholds were $300,000/QALY--a value higher than the $50,000 standard often cited. A subset of oncologists were sensitive to cost, believing it should factor into clinical decisions. These findings reflect the ongoing controversies within the medical community as expensive new therapies enter the system. PMID- 16476831 TI - The influence of endocrine effects of adjuvant therapy on quality of life outcomes in younger breast cancer survivors. AB - SIGNIFICANCE: There are 2.2 million breast cancer survivors, and approximately 25%-30% of newly diagnosed women each year are <50 years of age. Adjuvant therapy has prolonged survival, but the quality of that survival is influenced by persistent and late effects of therapy. Knowledge of treatment outcomes will assist in the design of interventions to prevent or manage persistent and late effects in survivors. PURPOSE: The purpose of this paper is to review the incidence of gonadal toxicity associated with adjuvant chemotherapy, side effects of endocrine therapy, quality of life outcomes, fertility concerns, and options to preserve fertility in young (<35 years) and young midlife (35-50 years) breast cancer survivors. RESULTS: Alkylating agent-based chemotherapy causes destruction of primordial follicles and impairment of follicular maturation resulting in temporary preservation of menses, reversible amenorrhea, irregular menses (perimenopause), or irreversible amenorrhea (ovarian failure--menopause). Younger women have a lower risk for amenorrhea with chemotherapy because of sufficient follicular stores, although the gonadal toxicity will result in an earlier than expected menopause. Premature menopause is associated with poorer quality of life, decreased sexual functioning, menopausal symptom distress, psychosocial distress related to fertility concerns, infertility, and uncertainty about late effects of premature menopause. Routine discussion about the menopausal experience, risks for infertility, and fertility preservation options is recommended. IMPLICATIONS FOR PRACTICE: This review identified adverse treatment outcomes for young and young midlife breast cancer survivors that can be minimized or prevented with targeted interventions. PMID- 16476832 TI - Breast cancer: achievements in adjuvant systemic therapies in the pre-genomic era. AB - In recent decades, the use of adjuvant systemic therapies for early breast cancer has increased extensively and has most likely contributed to the decline in breast cancer mortality observed in the U.S. and in some European countries. The last few years have witnessed accelerated progress in the treatment of early breast cancer, with the introduction of taxanes and aromatase inhibitors and, most impressively, trastuzumab to the adjuvant portfolio. When compared with anthracycline-based regimens, the addition of taxanes to treatments for patients with node-positive breast cancer has shown benefits in disease-free survival and, in some trials, in overall survival; however, these drugs are not yet universally accepted as standard treatment. Significant improvements in endocrine therapy in both pre- and postmenopausal patients with endocrine-responsive disease have been made. In the postmenopausal setting, aromatase inhibitors have shown superiority over tamoxifen in a direct comparison upfront or when given in sequence after 2-5 years of tamoxifen, but the optimal modality of administration remains unclear. For premenopausal women, ovarian function suppression with luteinizing hormone releasing hormone analogues combined with tamoxifen has generated similar results to cyclophosphamide, methotrexate, 5-fluorouracil (CMF)-based regimens. Recently, trastuzumab has had a dramatic impact on the evolution of human epidermal growth factor receptor 2 (HER-2)-positive early breast cancer treated with standard adjuvant modalities; specifically, relapses, including distant relapses, have been halved. In this review, we summarize these main achievements, discuss the currently available adjuvant treatment options for breast cancer patients, and emphasize the need for more efficient translational research to improve individual treatment tailoring. PMID- 16476834 TI - Postoperative chemotherapy for gastric cancer. AB - INTRODUCTION: Adjuvant chemotherapy for gastric cancer after potentially curative surgery has been under clinical investigation for more than four decades. However, potentially curative resection can be performed in only 30%-50% of patients. The objective of this article is to review briefly the clinical trials available in the current literature using adjuvant cytotoxic chemotherapy in patients with gastric cancer after potentially curative surgical resection. METHODS: Computerized (MEDLINE) and manual searches were performed to identify papers published on this topic between 1965 and 2005. Only articles with an English abstract were reviewed for inclusion; information abstracted included histologic proof of diagnosis, number of patients, dose and modality of treatment, survival duration, and side effects. RESULTS: Forty-three reports were identified. Single-agent chemotherapy was evaluated in four clinical trials, and postoperative combination chemotherapy was evaluated in 33 trials. Furthermore, we identified five meta-analyses. Five-year survival rates ranged from 12%-91.2%, and the median survival durations were 13-60+ months. Adjuvant chemotherapy, when compared with surgery alone, seems to result in longer survival. CONCLUSION: The high rate of recurrence, even in patients undergoing state-of-the art curative resection, suggests that effective adjuvant chemotherapy might indeed be an attractive concept to improve the overall outcome of patients with gastric cancer. However, because there is no standard regimen for postoperative treatment at the moment, patients with R0-resected (no residual tumors) gastric cancer should be offered the opportunity to participate in prospective clinical trials. PMID- 16476833 TI - Interethnic differences in genetic polymorphisms of CYP2D6 in the U.S. population: clinical implications. AB - DNA polymorphisms have been identified in the genes encoding a number of the cytochrome P450 (CYP) enzymes, leading to wide interindividual variation in drug clearance. CYP2D6 metabolizes a significant number of clinically used medications, and genetic variants of the CYP2D6 isozyme that result in varying levels of metabolic activity are of clinical importance in some settings. The exact nature of the clinical effect caused by polymorphisms of the gene depends on the drug in question and the specific variant alleles expressed, as individual variants result in differing phenotypes with a range of levels of enzymatic activity. Compromised drug efficacy due to CYP2D6 variation has been documented with a variety of agents, and this review considers a number of examples, including the 5-HT(3)-receptor antagonists, which are used in oncology supportive care for the prophylaxis of nausea and vomiting. CYP2D6 is involved in the metabolism of all of the most commonly available agents, except granisetron, and their efficacy and side effects may therefore be affected by the CYP2D6 polymorphism. Significant interethnic differences in CYP2D6 allele frequencies have been demonstrated from studies across many countries. However, incidences of polymorphisms in the U.S. population have been challenging to characterize because of the country's wide ethnic diversity. The CYP2D6 polymorphism may become more important as robust clinical tests become widely available and as the use of multiple medications and the attendant risk for drug-drug interactions increases. PMID- 16476835 TI - Alternating chemotherapy and radiotherapy in locally advanced head and neck cancer: an alternative? AB - Rapidly alternating chemotherapy and radiotherapy (ACR) is a minor variation of concurrent chemoradiation (CCR). This scheduling has been tested in advanced head and neck cancer and has shown superiority over standard radiation in some randomized trials with only marginally greater toxicity. This paper reviews ACR in advanced head and neck cancer. The hypothesis that this approach could have a better toxicity profile than CCR is discussed in light of the published clinical data. Efficacy is also discussed on the basis of available phase III trials. Published data indicate that rapidly alternating chemoradiation adds to toxicity less than CCR and results in comparable 3-year overall survival rates. In conclusion, ACR could be as active as, and possibly less toxic than, CCR. Comparative trials are highly recommended. PMID- 16476836 TI - Therapeutic advances in the treatment of glioblastoma: rationale and potential role of targeted agents. AB - Despite advances in standard therapy, including surgical resection followed by radiation and chemotherapy, the prognosis for patients with glioblastoma multiforme (GBM) remains poor. Unfortunately, most patients die within 2 years of diagnosis of their disease. Molecular abnormalities vary among individual patients and also within each tumor. Indeed, one of the distinguishing features of GBM is its marked genetic heterogeneity. Nonetheless, recent developments in the field of tumor biology have elucidated signaling pathways and genes involved in the development of GBM, and several novel agents that target these signaling pathways are being developed. As new details on the genetic characteristics of this disease become available, innovative treatment regimens, including a variety of traditional treatment modalities such as surgery, radiation, and cytotoxic chemotherapy, will be combined with newer targeted therapies. This review introduces these new targeted therapies in the context of current treatment options for patients with GBM. It is hoped that this combined approach will overcome the current limitations in the treatment of patients with GBM and result in a better prognosis for these patients. PMID- 16476837 TI - Changing paradigms--an update on the multidisciplinary management of malignant glioma. AB - Treatment of malignant glioma requires a multidisciplinary team. Treatment includes surgery, radiotherapy, and chemotherapy. Recently developed agents have demonstrated activity against recurrent malignant glioma and efficacy if given concurrently with radiotherapy in the upfront setting. Oligodendroglioma with 1p/19q deletions has been recognized as a distinct pathologic entity with particular sensitivity to radiotherapy and chemotherapy. Randomized trials have shown that early neoadjuvant or adjuvant administration of procarbazine, lomustine, and vincristine chemotherapy prolongs disease-free survival; however, it has no impact on overall survival. Temozolomide, a novel alkylating agent, has shown modest activity against recurrent glioma. In combination with radiotherapy in newly diagnosed patients with glioblastoma, temozolomide significantly prolongs survival. Molecular studies have demonstrated that the benefit is mainly observed in patients whose tumors have a methylated methylguanine methyltransferase gene promoter and are thus unable to repair some of the chemotherapy-induced DNA damage. For lower-grade glioma, the use of chemotherapy remains limited to recurrent disease, and first-line administration is the subject of ongoing clinical trials. Irinotecan and agents like gefitinib, erlotinib, and imatinib targeting the epidermal growth factor receptor and platelet-derived growth factor receptor have shown some promise in recurrent malignant glioma. This review summarizes recent developments, focusing on the clinical management of patients in daily neuro-oncology practice. PMID- 16476838 TI - Beta radiation. PMID- 16476839 TI - Cancer-related fatigue: predictors and effects of rehabilitation. AB - BACKGROUND: The aims of the study were to examine the effects of a multidimensional rehabilitation program on cancer-related fatigue, to examine concurrent predictors of fatigue, and to investigate whether change in fatigue over time was associated with change in predictors. METHODS: SAMPLE: 72 cancer survivors with different diagnoses. SETTING: rehabilitation center. INTERVENTION: 15-week rehabilitation program. MEASURES: Fatigue (Multidimensional Fatigue Inventory), demographic and disease/treatment-related variables, body composition (bioelectrical impedance), exercise capacity (symptom-limited bicycle ergometry), muscle force (handheld dynamometry), physical and psychological symptom distress (Rotterdam Symptom Check List), experienced physical and psychological functioning (RAND-36), and self-efficacy (General-Self-Efficacy Scale, Dutch version). Measurements were performed before (T0) and after rehabilitation (T1). RESULTS: At T1 (n = 56), significant improvements in fatigue were found, with effect sizes varying from -0.35 to -0.78. At T0, the different dimensions of fatigue were predicted by different physical and psychological variables. Explained variance of change in fatigue varied from 42%-58% and was associated with pre-existing fatigue and with change in physical functioning, role functioning due to physical problems, psychological functioning, and physical symptoms distress. CONCLUSIONS: Within this selected group of patients we found that (a) rehabilitation is effective in reducing fatigue, (b) both physical and psychological parameters predicted different dimensions of fatigue at baseline, and (c) change in fatigue was mainly associated with change in physical parameters. PMID- 16476841 TI - Early intervention with epoetin alfa during platinum-based chemotherapy: an analysis of the results of a multicenter, randomized, controlled trial based on initial hemoglobin level. AB - OBJECTIVE: This analysis of the results of a randomized, controlled trial evaluating the effects of epoetin alfa (EPO) therapy on transfusion requirements, hemoglobin (Hb), and quality of life (QOL) in patients with cancer receiving platinum-based chemotherapy was conducted to evaluate the effect of initial Hb level on study outcomes. METHODS: Patients with Hb levels < or =12.1 g/dl were randomized 2:1 to receive EPO, 10,000 U three times weekly s.c. or best supportive care (BSC) until 4 weeks after their last chemotherapy cycle. For this analysis, patients were stratified by baseline Hb level (< or =9.7 g/dl, >9.7 g/dl to < or =10.5 g/dl, >10.5 g/dl to < or =11.3 g/dl, and >11.3 g/dl to < or =12.1 g/dl), and study results were reanalyzed. RESULTS: Significantly fewer EPO patients than BSC patients with initial Hb levels >9.7 g/dl to < or =12.1 g/dl required transfusions. EPO maintained Hb levels throughout the study for patients with Hb levels >11.3 g/dl to < or =12.1 g/dl, compared with a decrease with BSC. For patients with baseline Hb levels >10.5 g/dl, for whom the mean changes from baseline to last assessment were measured by the Cancer Linear Analogue Scale assessments of energy and overall QOL as well as by the Functional Assessment of Cancer Therapy (FACT)-Fatigue and FACT-An Anemia subscale, QOL scores were significantly greater with EPO than with BSC. QOL declined in patients receiving BSC, and the mean decreases in QOL scores were greater for BSC patients with baseline Hb levels >10.5 g/dl, compared with the overall BSC group. CONCLUSION: In patients with cancer receiving platinum-based chemotherapy and with baseline Hb levels >10.5 g/dl, early intervention with EPO reduces transfusions, maintains Hb level, and maintains or improves QOL. This study supports the positive effects of early intervention when analyzed according to initial Hb value. PMID- 16476840 TI - Early Intervention with epoetin alfa during platinum-based chemotherapy: an analysis of quality-of-life results of a multicenter, randomized, controlled trial compared with population normative data. AB - OBJECTIVE: To evaluate the effect of epoetin alfa on quality of life (QOL) in patients with solid tumors and mild-to-moderate anemia receiving platinum-based chemotherapy relative to population norms. METHODS: In the original study, patients (n = 316) with hemoglobin (Hb) levels < or =12.1 g/dl were randomized 2:1 to receive either epoetin alfa at a dose of 10,000 U thrice weekly s.c. or best supportive care (BSC) to compare the effects on transfusion use, hematologic response, and QOL (measured by the Functional Assessment of Cancer Therapy-Anemia [FACT-An]and Cancer Linear Analogue Scale [CLAS]). The QOL data from this previously reported trial were reanalyzed here relative to population norms. RESULTS: Mean baseline QOL scores were similar between groups. At study completion, mean CLAS, FACT-An, FACT-An Anemia subscale, and FACT-An Fatigue subscale scores were significantly higher for patients given epoetin alfa than for those treated with BSC. Compared with population norms, both groups had impaired QOL at baseline. Differences in mean QOL change scores from baseline to study end for epoetin alfa versus BSC were 3.17 points for the FACT-General Total, 9.90 for the FACT-An Fatigue subscale, and 7.30 for the FACT-An Anemia subscale. This was equivalent to corrections in QOL deficits attributable to epoetin alfa of 97.3%, 40.7%, and 38.0% for the FACT-General Total, FACT-An Fatigue, and FACT-An Anemia subscale scores, respectively, versus BSC. A somewhat greater QOL benefit was observed for the FACT-An Fatigue and FACT-An Anemia subscales in the subset of patients with baseline Hb levels >10.5 g/dl. CONCLUSION: Patients in this study had impaired QOL compared with population norms. Early treatment with epoetin alfa to correct anemia improved QOL in a statistically significant and clinically meaningful way, and improvements were greater in patients with baseline Hb levels >10.5 g/dl. PMID- 16476842 TI - Direct-to-consumer advertising in oncology. AB - Shortly before his death in 1995, Kenneth B. Schwartz, a cancer patient at Massachusetts General Hospital (MGH), founded The Kenneth B. Schwartz Center at MGH. The Schwartz Center is a nonprofit organization dedicated to supporting and advancing compassionate health care delivery, which provides hope to patients and support to caregivers while encouraging the healing process. The center sponsors the Schwartz Center Rounds, a monthly multidisciplinary forum in which caregivers reflect on important psychosocial issues faced by patients, their families, and their caregivers, and gain insight and support from fellow staff members. Increasingly, cancer patients are subjected to advertisements related to oncologic therapies and other cancer-related products in the popular media. Such direct-to-consumer advertising is controversial: while it may inform, educate, and perhaps even empower patients, it also has the ability to misinform patients, and strain their relationships with oncology providers. The U.S. Food and Drug Administration requires that direct-to-consumer advertising provide a balanced presentation of a product's benefits, risks, and side effects, but this can be difficult to achieve. Through a discussion of this topic by an oncology fellow, ethicist, cancer survivor, and senior oncologist, the role of direct-to-consumer advertising and its often subtle effects on clinical practice in oncology are explored. Although sparse, the medical literature on this increasingly prevalent type of medical communication is also reviewed. PMID- 16476843 TI - Differential impact of blood pressure-lowering drugs on central aortic pressure and clinical outcomes: principal results of the Conduit Artery Function Evaluation (CAFE) study. AB - BACKGROUND: Different blood pressure (BP)-lowering drugs could have different effects on central aortic pressures and thus cardiovascular outcome despite similar effects on brachial BP. The Conduit Artery Function Evaluation (CAFE) study, a substudy of the Anglo-Scandinavian Cardiac Outcomes Trial (ASCOT), examined the impact of 2 different BP lowering-regimens (atenolol+/-thiazide based versus amlodipine+/-perindopril-based therapy) on derived central aortic pressures and hemodynamics. METHODS AND RESULTS: The CAFE study recruited 2199 patients in 5 ASCOT centers. Radial artery applanation tonometry and pulse wave analysis were used to derive central aortic pressures and hemodynamic indexes on repeated visits for up to 4 years. Most patients received combination therapy throughout the study. Despite similar brachial systolic BPs between treatment groups (Delta0.7 mm Hg; 95% CI, -0.4 to 1.7; P=0.2), there were substantial reductions in central aortic pressures with the amlodipine regimen (central aortic systolic BP, Delta4.3 mm Hg; 95% CI, 3.3 to 5.4; P<0.0001; central aortic pulse pressure, Delta3.0 mm Hg; 95% CI, 2.1 to 3.9; P<0.0001). Cox proportional hazards modeling showed that central pulse pressure was significantly associated with a post hoc-defined composite outcome of total cardiovascular events/procedures and development of renal impairment in the CAFE cohort (unadjusted, P<0.0001; adjusted for baseline variables, P<0.05). CONCLUSIONS: BP lowering drugs can have substantially different effects on central aortic pressures and hemodynamics despite a similar impact on brachial BP. Moreover, central aortic pulse pressure may be a determinant of clinical outcomes, and differences in central aortic pressures may be a potential mechanism to explain the different clinical outcomes between the 2 BP treatment arms in ASCOT. PMID- 16476844 TI - Postconditioning protects against endothelial ischemia-reperfusion injury in the human forearm. AB - BACKGROUND: Hypoxic cell death follows interruption of blood supply to tissues. Although successful restoration of blood flow is mandatory for salvage of ischemic tissues, reperfusion can paradoxically place tissues at risk of further injury. Brief periods of ischemia applied at the onset of reperfusion have been shown to reduce ischemia-reperfusion (IR) injury, a phenomenon called postconditioning. The aim of this study was to determine whether postconditioning protects against endothelial IR injury in humans, in vivo. METHODS AND RESULTS: Brachial artery endothelial function was assessed by vascular ultrasound to measure flow-mediated dilation (FMD) in response to forearm reactive hyperemia. FMD was measured before and after IR (20 minutes of arm ischemia followed by 20 minutes of reperfusion) in healthy volunteers. To test the protective effects of postconditioning, 3 cycles of reperfusion followed by ischemia (each lasting 10 or 30 seconds) were applied immediately after 20 minutes of arm ischemia. To determine whether postconditioning needs to be applied at the onset of reperfusion, a 1-minute period of arm reperfusion was allowed before the application of the 10-second postconditioning stimulus. IR caused endothelial dysfunction (FMD 9.1+/-1.2% pre-IR, 3.6+/-0.7% post-IR, P<0.001; n=11), which was prevented by postconditioning applied as 10-second cycles of reperfusion/ischemia (FMD 9.9+/-1.7% pre-IR, 8.3+/-1.4% post-IR, P=NS; n=11) and 30-second cycles of reperfusion/ischemia (FMD 10.8+/-1.7% pre-IR, 9.5+/-1.5% post-IR, P=NS; n=10) immediately at the onset of reperfusion. No protection was observed when the application of the 10-second postconditioning stimulus was delayed for 1 minute after the onset of reperfusion (FMD 9.8+/-1.2% pre-IR, 4.0+/-0.9% post-IR, P<0.001; n=8). CONCLUSIONS: This study demonstrates for the first time that postconditioning can protect against endothelial IR injury in humans. Postconditioning might reduce tissue injury when applied at the onset of reperfusion by modifying the reperfusion phase of IR. PMID- 16476845 TI - In vivo visualization of embryonic stem cell survival, proliferation, and migration after cardiac delivery. AB - BACKGROUND: Recent studies have shown that stem cell therapy can promote tissue regeneration; however, monitoring stem cells in vivo remains problematic owing to limitations of conventional histological assays and imaging modalities. METHODS AND RESULTS: Murine embryonic stem (ES) cells were stably transduced with a lentiviral vector carrying a novel triple-fusion (TF) reporter gene that consists of firefly luciferase, monomeric red fluorescence protein, and truncated thymidine kinase (fluc-mrfp-ttk). ES cell viability, proliferation, and differentiation ability were not adversely affected by either reporter genes or reporter probes compared with nontransduced control cells (P=NS). Afterward, 1x10(7) of ES cells carrying the TF reporter gene (ES-TF) were injected into the myocardium of adult nude rats (n=20). Control animals received nontransduced ES cells (n=6). At day 4, the bioluminescence and positron emission tomography signals in study animals were 3.7x10(7)+/-5.8x10(6) photons.s(-1).cm(-2) per steradian (sr) and 0.08+/-0.03% injected dose/g, respectively (P<0.05 versus control). Both signals increased progressively from week 1 to week 4, which indicated ES cell survival and proliferation in the host. Histological analysis demonstrated the formation of intracardiac and extracardiac teratomas. Finally, animals (n=4) that were treated with intraperitoneal injection of ganciclovir (50 mg/kg) did not develop teratomas when compared with control animals (n=4) treated with saline (1 mL/kg). CONCLUSIONS: This is the first study to characterize ES cells that stably express fluorescence, bioluminescence, and positron emission tomography reporter genes and monitor the kinetics of ES cell survival, proliferation, and migration. This versatile imaging platform should have broad applications for basic research and clinical studies on stem cell therapy. PMID- 16476846 TI - The presence of Lys27 instead of Asn27 in human phospholamban promotes sarcoplasmic reticulum Ca2+-ATPase superinhibition and cardiac remodeling. AB - BACKGROUND: Phospholamban (PLN) is an inhibitor of the Ca2+ affinity of sarcoplasmic reticulum (SR) Ca2+-ATPase (SERCA2). The amino acid sequence of PLN is highly conserved, and although all species contain asparagine (Asn), human PLN is unique in containing lysine (Lys) at amino acid 27. METHODS AND RESULTS: Human PLN was introduced in the null background. Expression of human PLN, at similar levels to mouse wild-type PLN, resulted in significant decreases in the affinity of SERCA2 for Ca2+, attributed to unique spatial conformation of this PLN form and increases in its monomeric active unit compared with mouse PLN. The increased inhibition by human PLN was associated with attenuated cardiac contractility in the intact-animal, organ, and cardiomyocyte levels and with depressed calcium kinetics. These inhibitory effects could not be fully reversed even on maximal isoproterenol stimulation. There were no alterations in the expression levels of SERCA2, calsequestrin, ryanodine receptor, and FKBP12, although the sodium/calcium exchanger and the L-type Ca2+ channel expression levels were upregulated. The depressed function resulted in increased heart/body weight ratios and phosphorylation levels of Akt, p38, and Erk1/2. CONCLUSIONS: Human PLN may play a more inhibitory role than that of other species in Ca2+ cycling. Expression of human PLN in the mouse is compensated by alterations in Ca2+ handling proteins and cardiac remodeling in an effort to normalize cardiac contractility. Thus, the unique amino acid sequence of human PLN may be critical in maintaining a high cardiac reserve, which is of paramount importance in the regulation of human cardiac function. PMID- 16476847 TI - Clinical correlates and consequences of anemia in a broad spectrum of patients with heart failure: results of the Candesartan in Heart Failure: Assessment of Reduction in Mortality and Morbidity (CHARM) Program. AB - BACKGROUND: We wished to determine the prevalence of, potential mechanistic associations of, and clinical outcomes related to anemia in patients with heart failure and a broad spectrum of left ventricular ejection fraction (LVEF). METHODS AND RESULTS: In multivariable analyses, we examined the associations between hemoglobin and baseline characteristics, laboratory variables, and outcomes in 2653 patients randomized in the CHARM Program in the United States and Canada. Anemia was equally common in patients with preserved (27%) and reduced (25%) LVEF but was more common in black and older patients. Anemia was associated with ethnicity, diabetes, low body mass index, higher systolic and lower diastolic blood pressure, and recent heart failure hospitalization. More than 50% of anemic patients had a glomerular filtration rate <60 mL.min(-1).1.73 m(-2) compared with <30% of nonanemic patients. Despite an inverse relationship between hemoglobin and LVEF, anemia was associated with an increased risk of death and hospitalization, a relationship observed in patients with both reduced and preserved LVEF. There were 133 versus 69 deaths and 527 versus 352 hospitalizations per 1000 patient-years of follow-up in anemic versus nonanemic patients (both P<0.001). The effect of candesartan in reducing outcomes was independent of hemoglobin. CONCLUSIONS: Anemia was common in heart failure, regardless of LVEF. Lower hemoglobin was associated with higher LVEF yet was an independent predictor of adverse mortality and morbidity outcomes. In heart failure, the causes of anemia and the associations between anemia and outcomes are probably multiple and complex. PMID- 16476848 TI - Cross-sectional relations of multiple biomarkers from distinct biological pathways to brachial artery endothelial function. AB - BACKGROUND: Endothelial dysfunction is a critical intermediate phenotype in the pathogenesis of cardiovascular disease. We evaluated the relative contributions of distinct biological pathways to interindividual variation in endothelial function by relating prototype biomarkers (representing these pathways) to brachial artery vasodilator function. METHODS AND RESULTS: We investigated the cross-sectional relations of a panel of 7 biomarkers measured at a routine examination to brachial artery vasodilator function (flow-mediated dilation [FMD] and reactive hyperemia) assessed at a subsequent examination (mean interval, 2.9 years) in 2113 Framingham Heart Study participants (mean age, 61 years; 54% women). We selected biomarkers from 4 biological domains: neurohormonal (N terminal pro-atrial natriuretic peptide [N-ANP], B-type natriuretic peptide [BNP], renin, aldosterone), hemostatic factors (plasminogen activator inhibitor-1 [PAI-1]), inflammation (C-reactive protein [CRP]), and target organ damage (urine albumin-creatinine ratio). In age- and sex-adjusted models, several biomarkers were related to baseline brachial artery diameter (PAI-1, CRP, urine albumin creatinine ratio), baseline mean flow (N-ANP, BNP, PAI-1, CRP, aldosterone), FMD (N-ANP, PAI-1, CRP, renin), and reactive hyperemia (BNP, PAI-1, CRP, renin, urine albumin-creatinine ratio). In multivariable analyses relating the 7 biomarkers conjointly to each vascular function measure (adjusting for known risk factors), N-ANP and renin were positively related to FMD (P=0.001 and P=0.04, respectively), and N-ANP was inversely related to baseline mean flow velocity (P=0.01). None of the other biomarkers was significantly related to the vascular function measures studied. CONCLUSIONS: In our large community-based sample, a conservative strategy relating several biomarkers to vascular endothelial function identified plasma N-ANP as a key correlate of mean flow under basal conditions and of FMD in response to forearm cuff occlusion. PMID- 16476849 TI - Biventricular pacing improves the blunted force-frequency relation present during univentricular pacing in patients with heart failure and conduction delay. AB - BACKGROUND: In patients with chronic heart failure (CHF) and conduction delay, biventricular (BiV) and left ventricular (LV) pacing similarly improve systolic function at resting heart rates. We hypothesized that BiV and univentricular pacing differentially affect contractile function at increasing heart rates. METHODS AND RESULTS: Twenty-two patients (aged 66+/-2 years, QRS 179+/-8 ms, LV ejection fraction 23+/-1%) underwent cardiac catheterization before device implantation to measure LV hemodynamics at baseline (rate 68+/-2 bpm; sinus rhythm n=18; atrial fibrillation n=4) and during BiV, LV, and right ventricular (RV) stimulation at 80, 100, 120, and 140 bpm. BiV and LV pacing at 80 bpm equally augmented dP/dtmax as compared with baseline and RV pacing (P<0.001). Stimulation rate significantly interacted with the effect of BiV, LV, and RV pacing on LV end-diastolic pressure (LVEDP), systolic pressure (LVSP), and dP/dtmax. Increasing the rate from 80 to 140 bpm enhanced dP/dtmax from 913+/-28 to 1119+/-50 mm Hg/s during BiV stimulation (P<0.001) but had no significant effect on contractility during single-site LV (951+/-47 versus 1002+/-54 mm Hg/s) or RV (800+/-46 versus 881+/-49 mm Hg/s) pacing. At 140 bpm, LVEDP was lower and LVSP higher during BiV pacing than during RV and LV pacing (LVEDP 12+/-1 versus 17+/-1 and 16+/-1 mm Hg, P<0.001; LVSP 112+/-5 versus 106+/-5 and 108+/-6 mm Hg, P<0.01 and P=0.09; BiV versus RV and LV pacing, respectively). CONCLUSIONS: Different modes of ventricular stimulation alter the in vivo force-frequency relation of CHF patients. In contrast to single-site LV and RV pacing, contractile function improves with increasing heart rates during BiV stimulation. This effect may contribute to the enhanced exercise capacity during BiV pacing and could provide a functional benefit over LV-only pacing in patients for whom resynchronization therapy is indicated. PMID- 16476850 TI - Novel speckle-tracking radial strain from routine black-and-white echocardiographic images to quantify dyssynchrony and predict response to cardiac resynchronization therapy. AB - BACKGROUND: Mechanical dyssynchrony is a potential means to predict response to cardiac resynchronization therapy (CRT). We hypothesized that novel echocardiographic image speckle tracking can quantify dyssynchrony and predict response to CRT. METHODS AND RESULTS: Seventy-four subjects were studied: 64 heart failure patients undergoing CRT (aged 64+/-12 years, ejection fraction 26+/ 6%, QRS duration 157+/-28 ms) and 10 normal controls. Speckle tracking applied to routine midventricular short-axis images calculated radial strain from multiple circumferential points averaged to 6 standard segments. Dyssynchrony from timing of speckle-tracking peak radial strain was correlated with tissue Doppler measures in 47 subjects (r=0.94, P<0.001; 95% CI 0.90 to 0.96). The ability of baseline speckle-tracking radial dyssynchrony (time difference in peak septal wall-to-posterior wall strain > or =130 ms) to predict response to CRT was then tested. It predicted an immediate increase in stroke volume in 48 patients studied the day after CRT with 91% sensitivity and 75% specificity. In 50 patients with long-term follow-up 8+/-5 months after CRT, baseline speckle tracking radial dyssynchrony predicted a significant increase in ejection fraction with 89% sensitivity and 83% specificity. Patients in whom left ventricular lead position was concordant with the site of latest mechanical activation by speckle-tracking radial strain had an increase in ejection fraction from baseline to a greater degree (10+/-5%) than patients with discordant lead position (6+/-5%; P<0.05). CONCLUSIONS: Speckle-tracking radial strain can quantify dyssynchrony and predict immediate and long-term response to CRT and has potential for clinical application. PMID- 16476851 TI - Introduction of metoprolol increases plasma B-type cardiac natriuretic peptides in mild, stable heart failure. AB - BACKGROUND: The effect of beta-blockade on the cardiac natriuretic peptides is poorly understood but could contribute to their beneficial treatment effect and may be relevant to clinical use of plasma brain natriuretic peptide (BNP)/N terminal pro brain natriuretic peptide (NTproBNP) measurements in risk stratification and in titration of anti-heart failure therapy. METHODS AND RESULTS: Sixteen men with mild, stable heart failure (NYHA class II to III; left ventricular ejection fraction <40%) underwent serial blood sampling for plasma natriuretic peptide levels and received infusions of atrial natriuretic peptide (ANP) and BNP before and 6 weeks after the introduction and uptitration of metoprolol or 6 weeks unchanged therapy in a randomized, parallel-group design. Plasma natriuretic peptides (BNP, NTproBNP, ANP, and NTproANP) were increased by metoprolol (P<0.01 for all). The natriuretic responses to ANP and BNP infusions were sustained with the introduction of metoprolol despite reduced renal perfusion pressure. The levels of the noninfused natriuretic peptide were increased by both ANP and BNP infusions, and this effect was enhanced by metoprolol. The early plasma half-life (t(1/2)alpha) of BNP was prolonged by metoprolol (5.6+/-0.45 to 11+/-1.3 versus 5.7+/-0.8 to 6.6+/-1.3 minutes in control subjects; P=0.019). CONCLUSIONS: Plasma cardiac natriuretic peptide levels increase significantly with the introduction of metoprolol in heart failure as a result of effects on secretion and clearance. Natriuretic responses to NP infusions are sustained with beta-blockade despite reduced renal perfusion pressure. Clinicians should be aware that the introduction of metoprolol causes a rise in plasma BNP/NTproBNP that is unrelated to deterioration in clinical status and must be considered when measurements are undertaken for risk stratification or titration of treatment. PMID- 16476852 TI - Effect of posterolateral scar tissue on clinical and echocardiographic improvement after cardiac resynchronization therapy. AB - BACKGROUND: Currently, one third of patients treated with cardiac resynchronization therapy (CRT) do not respond. Nonresponse to CRT may be explained by the presence of scar tissue in the posterolateral left ventricular (LV) segments, which may result in ineffective LV pacing and inadequate LV resynchronization. In the present study, the relationship between transmural posterolateral scar tissue and response to CRT was evaluated. METHODS AND RESULTS: Forty consecutive patients with end-stage heart failure (NYHA class III/IV), LV ejection fraction < or =35%, QRS duration >120 ms, left bundle-branch block, and chronic coronary artery disease were included. The localization and transmurality of scar tissue were evaluated with contrast-enhanced MRI. Next, LV dyssynchrony was assessed at baseline and immediately after implantation with tissue Doppler imaging. Clinical parameters, LV volumes, and LV ejection fraction were assessed at baseline and at a 6-month follow-up. Fourteen patients (35%) had a transmural (>50% of LV wall thickness) posterolateral scar. In contrast to patients without posterolateral scar tissue, these patients showed a low response rate (14% versus 81%; P<0.05) and did not show improvement in clinical or echocardiographic parameters. In addition, LV dyssynchrony remained unchanged after CRT implantation (84+/-46 versus 78+/-41 ms; P=NS). Patients without posterolateral scar tissue and severe baseline dyssynchrony (> or =65 ms) showed an excellent response rate of 95% compared with patients with a posterolateral scar and/or absent LV dyssynchrony (11%). CONCLUSIONS: CRT does not reduce LV dyssynchrony in patients with transmural scar tissue in the posterolateral LV segments, resulting in clinical and echocardiographic nonresponse to CRT. PMID- 16476853 TI - Inflammatory biomarkers in acute coronary syndromes: part I: introduction and cytokines. PMID- 16476854 TI - Images in cardiovascular medicine. Disruption of dystrophin in acute fulminant coxsackieviral B4 infection. PMID- 16476855 TI - Images in cardiovascular medicine. Assessment of isolated right ventricular myocardial infarction by magnetic resonance imaging. PMID- 16476856 TI - Images in cardiovascular medicine. Multiple thrombosis with aortic occlusion after heparin-induced thrombocytopenia demonstrated by multislice computed tomography. PMID- 16476857 TI - A view from Ankara. PMID- 16476858 TI - Use and misuse of surrogate outcomes in arrhythmia trials. PMID- 16476859 TI - Continuous quality improvement in the cardiac catheterization laboratory: are the benefits worth the cost and effort? PMID- 16476860 TI - Percutaneous valve repair and replacement: challenges encountered, challenges met, challenges ahead. PMID- 16476862 TI - Myocarditis: current trends in diagnosis and treatment. PMID- 16476861 TI - Percutaneous valve intervention: a surgeon's perspective. PMID- 16476863 TI - Postmarket surveillance for drug-eluting coronary stents: a comprehensive approach. PMID- 16476864 TI - NO in early pregnancy and development of preeclampsia. PMID- 16476865 TI - Screening for hemochromatosis. PMID- 16476866 TI - Universal patient disinfection as a tool for infection control: rub-a-dub-dub, no need for a tub. PMID- 16476867 TI - An American physician in the Canadian health care system. PMID- 16476868 TI - Effects of low-carbohydrate vs low-fat diets on weight loss and cardiovascular risk factors: a meta-analysis of randomized controlled trials. AB - BACKGROUND: Low-carbohydrate diets have become increasingly popular for weight loss. However, evidence from individual trials about benefits and risks of these diets to achieve weight loss and modify cardiovascular risk factors is preliminary. METHODS: We used the Cochrane Collaboration search strategy to identify trials comparing the effects of low-carbohydrate diets without restriction of energy intake vs low-fat diets in individuals with a body mass index (calculated as weight in kilograms divided by the square of height in meters) of at least 25. Included trials had to report changes in body weight in intention-to-treat analysis and to have a follow-up of at least 6 months. Two reviewers independently assessed trial eligibility and quality of randomized controlled trials. RESULTS: Five trials including a total of 447 individuals fulfilled our inclusion criteria. After 6 months, individuals assigned to low carbohydrate diets had lost more weight than individuals randomized to low-fat diets (weighted mean difference, -3.3 kg; 95% confidence interval [CI], -5.3 to 1.4 kg). This difference was no longer obvious after 12 months (weighted mean difference, -1.0 kg; 95% CI, -3.5 to 1.5 kg). There were no differences in blood pressure. Triglyceride and high-density lipoprotein cholesterol values changed more favorably in individuals assigned to low-carbohydrate diets (after 6 months, for triglycerides, weighted mean difference, -22.1 mg/dL [-0.25 mmol/L]; 95% CI, 38.1 to -5.3 mg/dL [-0.43 to -0.06 mmol/L]; and for high-density lipoprotein cholesterol, weighted mean difference, 4.6 mg/dL [0.12 mmol/L]; 95% CI, 1.5-8.1 mg/dL [0.04-0.21 mmol/L]), but total cholesterol and low-density lipoprotein cholesterol values changed more favorably in individuals assigned to low-fat diets (weighted mean difference in low-density lipoprotein cholesterol after 6 months, 5.4 mg/dL [0.14 mmol/L]; 95% CI, 1.2-10.1 mg/dL [0.03-0.26 mmol/L]). CONCLUSIONS: Low-carbohydrate, non-energy-restricted diets appear to be at least as effective as low-fat, energy-restricted diets in inducing weight loss for up to 1 year. However, potential favorable changes in triglyceride and high-density lipoprotein cholesterol values should be weighed against potential unfavorable changes in low-density lipoprotein cholesterol values when low-carbohydrate diets to induce weight loss are considered. PMID- 16476869 TI - Screening for hemochromatosis in asymptomatic subjects with or without a family history. AB - BACKGROUND: Hemochromatosis in white subjects is mostly due to homozygosity for the common C282Y substitution in HFE. Although clinical symptoms are preventable by early detection of the genetic predisposition and prophylactic treatment, population screening is not currently advocated because of the discrepancy between the common mutation prevalence and apparently lower frequency of clinical disease. This study compared screening for hemochromatosis in subjects with or without a family history. METHODS: We assessed disease expression by clinical evaluation and liver biopsy in 672 essentially asymptomatic C282Y homozygous subjects identified by either family screening or health checks. We also observed a subgroup of untreated homozygotes with normal serum ferritin levels for up to 24 years. RESULTS: Prevalence of hepatic iron overload and fibrosis were comparable between the 2 groups. Disease-related conditions were more common in male subjects identified by health checks, but they were older. Hepatic iron overload (grades 2-4) was present in 56% and 34.5% of male and female subjects, respectively; hepatic fibrosis (stages 2-4) in 18.4% and 5.4%; and cirrhosis in 5.6% and 1.9%. Hepatic fibrosis and cirrhosis correlated significantly with the hepatic iron concentration, and except in cases of cirrhosis, there was a 7.5 fold reduction in the mean fibrosis score after phlebotomy. All subjects with cirrhosis were asymptomatic. CONCLUSIONS: Screening for hemochromatosis in apparently healthy subjects homozygous for the C282Y mutation with or without a family history reveals comparable levels of hepatic iron overload and disease. Significant hepatic fibrosis is frequently found in asymptomatic subjects with hemochromatosis and, except when cirrhosis is present, is reversed by iron removal. PMID- 16476871 TI - Utility of amino-terminal pro-brain natriuretic peptide testing for prediction of 1-year mortality in patients with dyspnea treated in the emergency department. AB - BACKGROUND: Amino-terminal pro-brain natriuretic peptide (NT-proBNP) is useful for diagnosis and triage of patients with dyspnea, but its role for predicting outcomes in such patients remains undefined. METHODS: A total of 599 breathless patients treated in the emergency department were prospectively enrolled, and a sample of blood was obtained for NT-proBNP measurements. After 1 year, the vital status of each patient was ascertained, and the association between NT-proBNP values at presentation and mortality was assessed. RESULTS: At 1 year, 91 patients (15.2%) had died. Median NT-proBNP concentrations at presentation among decedents were significantly higher than those of survivors (3277 vs 299 pg/mL; P<.001). The optimal NT-proBNP cut point for predicting 1-year mortality was 986 pg/mL. In a multivariable model, an NT-proBNP concentration greater than 986 pg/mL at presentation was the single strongest predictor of death at 1 year (hazard ratio [HR], 2.88; 95% confidence interval, 1.64-5.06; P<.001), independent of a diagnosis of heart failure. Other factors associated with death included age (by decade; HR, 1.20), heart rate (by decile; HR, 1.13), urea nitrogen level (by decile; HR, 1.20), systolic blood pressure less than 100 mm Hg (HR, 1.94), heart murmur (HR, 1.92), and New York Heart Association classification (HR, 1.38 for each increase in class). The NT-proBNP concentration alone had an area under the receiver operating characteristic curve (AUC) of 0.76 for predicting mortality; the other significant covariates combined had an AUC of 0.80. The final model for predicting death, combining NT-proBNP with other covariates associated with mortality, had a superior AUC of 0.82. CONCLUSION: In addition to assisting in emergency department diagnosis and triage, NT-proBNP concentrations at presentation are strongly predictive of 1-year mortality in dyspneic patients. PMID- 16476870 TI - Chlorhexidine gluconate to cleanse patients in a medical intensive care unit: the effectiveness of source control to reduce the bioburden of vancomycin-resistant enterococci. AB - BACKGROUND: Historically, methods of interrupting pathogen transmission have focused on improving health care workers' adherence to recommended infection control practices. An adjunctive approach may be to use source control (eg, to decontaminate patients' skin). METHODS: We performed a prospective sequential group single-arm clinical trial in a teaching hospital's medical intensive care unit from October 2002 to December 2003. We bathed or cleansed 1787 patients and assessed them for acquisition of vancomycin-resistant enterococci (VRE). We performed a nested study of 86 patients with VRE colonization and obtained culture specimens from 758 environmental surfaces and 529 health care workers' hands. All patients were cleansed daily with the procedure specific to the study period as follows: period 1, soap and water baths; period 2, cleansing with cloths saturated with 2% chlorhexidine gluconate; and period 3, cloth cleansing without chlorhexidine. We measured colonization of patient skin by VRE, health care worker hand or environmental surface contamination by VRE, and patient acquisition of VRE rectal colonization. RESULTS: Compared with soap and water baths, cleansing patients with chlorhexidine-saturated cloths resulted in 2.5 log(10) less colonies of VRE on patients' skin and less VRE contamination of health care workers' hands (risk ratio [RR], 0.6; 95% confidence interval [CI], 0.4-0.8) and environmental surfaces (RR, 0.3; 95% CI, 0.2-0.5). The incidence of VRE acquisition decreased from 26 colonizations per 1000 patient-days to 9 per 1000 patient-days (RR, 0.4; 95% CI, 0.1-0.9). For all measures, effectiveness of cleansing with nonmedicated cloths was similar to that of soap and water baths. CONCLUSION: Cleansing patients with chlorhexidine-saturated cloths is a simple, effective strategy to reduce VRE contamination of patients' skin, the environment, and health care workers' hands and to decrease patient acquisition of VRE. PMID- 16476872 TI - Computed tomographic screening for lung cancer: the relationship of disease stage to tumor size. AB - BACKGROUND: The relationship of lung cancer stage to tumor diameter has been identified as a prognostic indicator. We report on the stage-size relationship of these asymptomatic, latent lung cancer cases diagnosed by computed tomographic screening. METHODS: Baseline and repeat screening of 28 689 people following the International Early Lung Cancer Action Program regimen of screening has resulted in 464 diagnoses of lung cancer. Each case was characterized according to tumor diameter, consistency (solid, part solid, or nonsolid), and the presence or absence of identifiable metastases (N0 M0) at the time of diagnosis, regardless of whether it was delayed. RESULTS: For the 436 non-small cell carcinomas, the percentages of cases with no metastases (N0 M0) were 91%, 83%, 68%, and 55% for the categories 15 mm or less, 16 to 25 mm, 26 to 35 mm, and 36 mm or greater, respectively. The gradients in the successive percentages of N0 M0 cases were significantly different (P = .02, 1-sided), except between the last 2 categories, and held for solid nodules, were suggestive for part-solid ones, but were not suggestive for nonsolid ones. For the 28 small cell carcinomas, the percentages of N0 M0 cases were 67% and 23% (P = .01, 1-sided), respectively, for those 25 mm or less compared with those greater than 25 mm. CONCLUSIONS: Lymph node status has a strong relationship to tumor diameter for non-small cell and small cell cancers. The percentages of N0 M0 cases in screen-diagnosed lung cancers are much higher than previously reported in the Surveillance, Epidemiology, and End Results registry. These results provide direct evidence of a stage-size relationship in a screened population. PMID- 16476873 TI - Differences in health care utilization at the end of life among patients with chronic obstructive pulmonary disease and patients with lung cancer. AB - BACKGROUND: We sought to examine health care resource utilization in the last 6 months of life among patients who died with chronic obstructive pulmonary disease (COPD) compared with those who died with lung cancer and to examine geographic variations in care. METHODS: We performed a retrospective cohort study of patients diagnosed as having COPD or lung cancer, who were seen in 1 of 7 Veteran Affairs medical centers primary care clinics and who died during the study period. Our outcome of interest was health care resource utilization in the last 6 months of life. RESULTS: In the last 6 months of life, patients with COPD were more likely to visit their primary care providers but had fewer hospital admissions compared with patients with lung cancer. Patients with COPD had twice the odds of being admitted to an intensive care unit (ICU), 5 times the odds of remaining there 2 weeks or longer, and received fewer opiates and benzodiazepine prescriptions compared with patients with lung cancer. There were geographic variations in the use of ICUs for patients with COPD but not for those with lung cancer. Total health care costs were $4000 higher for patients with COPD because of ICU utilization. CONCLUSIONS: In the last 6 months of life, patients with COPD were more likely to have had a primary care visit and been admitted to an ICU but less likely to receive palliative medications compared with patients with lung cancer. We found significant geographic variability in ICU utilization but only for patients with COPD. PMID- 16476874 TI - The implications of choice: prescribing generic or preferred pharmaceuticals improves medication adherence for chronic conditions. AB - BACKGROUND: A large proportion of Americans are enrolled in 3-tier pharmacy benefit plans. We studied whether patients enrolled in such plans who receive generic or preferred brand-name agents when initiating chronic therapy were more adherent to treatment than those who received nonpreferred brand-name medications. METHODS: We analyzed pharmacy claims filled between October 1, 2001, and October 1, 2003, from a large health plan for 6 classes of chronic medications: 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, calcium channel blockers, oral contraceptives, orally inhaled corticosteroids, angiotensin receptor blockers, and angiotensin-converting enzyme inhibitors. We measured adherence as the proportion of days covered (PDC) in each drug class during the first year of therapy. We evaluated how the formulary status of the initial prescription (generic, preferred, or nonpreferred) influenced PDC and adequate adherence, defined as PDC greater than 80%, over the subsequent year. RESULTS: A total of 7532 new prescriptions were filled in 1 of the classes evaluated: 1747 (23.2%) for nonpreferred medications, 4376 (58.1%) for preferred drugs, and 1409 (18.7%) for generic drugs. After controlling for patient sociodemographic characteristics and drug class, PDC was 12.6% greater for patients initiated on generic medications vs nonpreferred medications (58.8% vs 52.2%; P<.001). The PDC was 8.8% greater for patients initiated on preferred vs nonpreferred medications (56.8% vs 52.2%; P<.001). Patients initiated on generic and preferred medications had 62% and 30% greater odds, respectively, of achieving adequate adherence compared with those who received nonpreferred medications. CONCLUSION: In 3-tier pharmacy benefit plans, prescribing generic or preferred medications within a therapeutic class is associated with improvements in adherence to therapy. PMID- 16476875 TI - Adherence to black box warnings for prescription medications in outpatients. AB - BACKGROUND: Few data are available regarding the prevalence of potentially dangerous drug-drug, drug-laboratory, and drug-disease interactions among outpatients. Our objectives were to determine how frequently clinicians prescribe drugs in violation of black box warnings for these issues and to determine how frequently such prescribing results in harm. METHODS: In an observational study of 51 outpatient practices using an electronic health record, we measured the frequency with which patients received prescriptions in violation of black box warnings for drug-drug, drug-laboratory, and/or drug-disease interactions. We performed medical record reviews in a sample of patients to detect adverse drug events. Multivariate analysis was conducted to assess the relationship of prescribing in violation of black box warnings to patient and clinician characteristics, adjusting for potential confounders and clustering. RESULTS: Of 324 548 outpatients who received a medication in 2002, 2354 (0.7%) received a prescription in violation of a black box warning. After adjustment, receipt of medication in violation of a black box warning was more likely when patients were 75 years or older or female. The number of medications taken, the number of medical problems, and the site of care were also associated with violations. Less than 1% of patients who received a drug in violation of a black box warning had an adverse drug event as a result. CONCLUSIONS: About 7 in 1000 outpatients received a prescription violating a black box warning. Few incidents resulted in detectable harm. PMID- 16476876 TI - C-reactive protein concentration and incident hypertension in young adults: the CARDIA study. AB - BACKGROUND: There is increasing evidence that C-reactive protein (CRP) concentration, a measure of inflammation, is an independent risk factor for the development of hypertension in older adults. However, it is unknown whether a similar relationship exists in younger individuals. METHODS: The Coronary Artery Risk Development in Young Adults (CARDIA) study was initiated in 1985-1986 to determine the factors that are associated with coronary risk development in young adults. C-reactive protein concentrations were measured in 3919 African American and white men and women enrolled in CARDIA using blood specimens from the year 7 examination (1992-1993), when the age of the cohort was 25 to 37 years, and the year 15 examination (2000-2001). RESULTS: In unadjusted analyses, CRP concentrations greater than 3 mg/L, compared with those less than 1 mg/L, was associated with a 79% greater risk of incident hypertension (odds ratio [OR], 1.79; 95% confidence interval [CI], 1.40-2.28). However, CRP concentration did not predict risk of incident hypertension after adjusting for year 7 body mass index (BMI) (OR, 1.14; 95% CI, 0.86-1.53) or year 7 BMI and other potential confounders (OR, 1.13; 95% CI, 0.83-1.52). In addition, year 7 CRP concentration was not associated with change in systolic or diastolic blood pressure after adjusting for BMI (P = .10 and P = .70, respectively). These findings were similar within each of the race- and sex-specific groups. CONCLUSION: C-reactive protein is associated with hypertension in young adults, but in contrast to the finding in older populations, the association is no longer present after adjusting for BMI. PMID- 16476878 TI - Conjugated equine estrogens and coronary heart disease: the Women's Health Initiative. AB - BACKGROUND: In recent randomized trials, conjugated equine estrogens (CEE) with continuous medroxyprogesterone acetate provided no protection against coronary heart disease in postmenopausal women and may have increased cardiac risk. These trials did not address the role of unopposed estrogen for coronary protection. METHODS: A total of 10 739 women aged 50 to 79 years at baseline (mean age, 63.6 years) who had previously undergone hysterectomy were randomized to receive CEE, 0.625 mg/d, or placebo at 40 US clinical centers beginning in 1993. The trial was terminated early after 6.8 years of follow-up (planned duration, 8.5 years). This report includes final, centrally adjudicated results for the primary efficacy outcome (myocardial infarction or coronary death), secondary coronary outcomes, and subgroup analyses. RESULTS: During the active intervention period, 201 coronary events were confirmed among women assigned to receive CEE compared with 217 events among women assigned to receive placebo (hazard ratio, 0.95; nominal 95% confidence interval, 0.79-1.16). Among women aged 50 to 59 years at baseline, the hazard ratio for the primary outcome was 0.63 (nominal 95% confidence interval, 0.36-1.08). In that age group, coronary revascularization was less frequent among women assigned to receive CEE (hazard ratio, 0.55; nominal 95% confidence interval, 0.35-0.86), as were several composite outcomes, which included the primary outcome and coronary revascularization (hazard ratio, 0.66; nominal 95% confidence interval, 0.44-0.97). CONCLUSIONS: Conjugated equine estrogens provided no overall protection against myocardial infarction or coronary death in generally healthy postmenopausal women during a 7-year period of use. There was a suggestion of lower coronary heart disease risk with CEE among women 50 to 59 years of age at baseline. PMID- 16476879 TI - C-reactive protein and vascular cell adhesion molecule-1 as markers of severity in sickle cell disease. PMID- 16476877 TI - Adipocytokines and incident diabetes mellitus in older adults: the independent effect of plasminogen activator inhibitor 1. AB - BACKGROUND: Adipose tissue produces "adipocytokines" of uncertain clinical significance. METHODS: We analyzed the relationships among adiposity, adipocytokines, glycemia, and incident diabetes mellitus in 2356 white and black adults aged 70 to 79 years in the Health, Aging, and Body Composition Study who did not have diabetes at baseline. We measured the levels of adipocytokines adiponectin, leptin, interleukin 6, tumor necrosis factor alpha, and plasminogen activator inhibitor 1. Regional fat area was determined by means of computed tomography. New diabetes was defined as a self-reported diagnosis of diabetes or as a fasting plasma glucose level of 126 mg/dL or greater (>/=7.0 mmol/L) at the second, fourth, or sixth annual examination. RESULTS: A total of 143 participants (14.1 cases per 1000 person-years) developed diabetes across 5 years. Visceral fat area (odds ratio [OR], 1.33; 95% confidence interval [CI], 1.10-1.60 per standard deviation increase) and body mass index (white individuals: OR, 1.65; 95% CI, 1.26-2.15 per standard deviation increase; black individuals: OR, 1.22; 95% CI, 0.99-1.51 per standard deviation increase) independently predicted incident diabetes. Adiponectin, leptin, and plasminogen activator inhibitor 1 attenuated the relationship between adiposity and diabetes. After controlling for body mass index, visceral fat, fasting glucose, fasting insulin, high-density lipoprotein cholesterol, triglycerides, and hypertension at baseline, plasminogen activator inhibitor 1 was the only adipocytokine independently associated with increased odds of diabetes (OR, 1.35; 95% CI, 1.01-1.81). Fasting glucose level at baseline remained a strong predictor of incident diabetes, whereas associations with body mass index and visceral fat were attenuated. CONCLUSIONS: Adipocytokines and glycemia partially account for the relationship between adiposity and risk of type 2 diabetes due to adiposity. Plasminogen activator inhibitor 1 may be a useful predictor of diabetes in addition to measurements of body fat. PMID- 16476880 TI - Omission of drug dose information. PMID- 16476881 TI - The annual examination: a touchstone for contextualized care. PMID- 16476882 TI - A change of reason: medicine and the scientific revolution. PMID- 16476883 TI - Studies on natural products. PMID- 16476884 TI - Increase of peak intraocular pressure during sleep in reproduced diurnal changes by posture. AB - OBJECTIVE: To characterize diurnal intraocular pressure (IOP) changes in primary open-angle glaucoma by reproducing IOPs based on patient posture. METHODS: In 148 patients with untreated primary open-angle glaucoma who had IOPs recorded during clinic hours that were less than 21 mm Hg (average, 14.8 +/- 3.2 mm Hg), we measured IOP by noncontact tonometry every 2 hours from 6 am to midnight and every 3 hours from midnight to 6 am with patients sitting and supine. The IOP was reproduced by designating the sitting IOP as measurements taken when the patient was awake and the supine IOP as measurements taken when the patient was asleep for each individual. The reproduced diurnal IOP was composed of 12 measurements that included 2 to 4 IOP levels measured with the patients supine and the rest while they were sitting. RESULTS: The peak of sitting diurnal IOP (mean +/- SD) for 148 patients was 16.0 +/- 2.7 mm Hg, which was significantly lower than the peak of supine IOP (18.9 +/- 3.9 mm Hg) or the reproduced IOP (17.5 +/- 3.6 mm Hg) (P<.001 for both comparisons). The average reproduced IOP at each measurement time peaked at 3 am during sleep; with sitting diurnal IOP or supine diurnal IOP, the peak IOPs were at noon. Twenty-nine patients (20%) with an IOP less than 21 mm Hg during clinic hours had a reproduced IOP of 21 mm Hg or greater while asleep, compared with only 5 patients (3%) when the patients were sitting only. CONCLUSIONS: In patients with primary open-angle glaucoma and IOPs less than 21 mm Hg during clinic hours, 20% of patients had a reproduced IOP of 21 mm Hg or greater, compared with only 3% who had an IOP of 21 mm Hg or greater while sitting. Intraocular pressures peaked in most patients during sleep. PMID- 16476885 TI - Detection of psychophysical and structural injury in eyes with glaucomatous optic neuropathy and normal standard automated perimetry. AB - OBJECTIVE: To compare the prevalence of structural and psychophysical abnormalities in normal eyes and eyes with glaucomatous optic neuropathy (GON) and normal standard automated perimetry (SAP). METHODS: Complete examination, SAP, short-wavelength automated perimetry (SWAP), frequency doubling technology (FDT), scanning laser polarimetry (GDx-VCC), and optical coherence tomography (OCT) of the peripapillary retinal nerve fiber layer (RNFL), optic disc, and macula were performed. Glaucomatous optic neuropathy was defined as cup-disc asymmetry between fellow eyes of greater than 0.2, rim thinning, notching, excavation, or RNFL defect. All eyes had normal SAP. Abnormal measurements on OCT, GDx-VCC, SWAP, and FDT were defined as those outside 95% normal limits. Eyes were stratified into 3 groups based on the OCT-generated vertical cup-disc ratio: mild, moderate, and advanced cupping (cup-disc ratio of <0.4, 0.4-0.7, and >0.7, respectively). Receiver operating characteristic curves were developed to assess sensitivity and specificity of structural and functional assessments. RESULTS: Forty-seven eyes of 47 patients (25 with GON and 22 normal) were enrolled (mean +/- SD age, 58 +/- 16 years; range, 25-83 years). Compared with normal eyes, eyes with GON had significantly worse mean deviation and pattern standard deviation by means of SWAP and FDT (P = .02-.05); OCT-derived mean and superior and inferior RNFL thickness (P = .008, <.001, and .05, respectively); mean macular thickness (P = .01), rim volume, rim area, cup-disc ratio, and cup area (all P<.001); and GDx-VCC nerve fiber indicator and inferior average (P = .03). There was a significantly (P = .008, .002, .003, and .01, respectively) greater prevalence of abnormalities identified by SWAP, FDT, OCT and GDx-VCC in eyes with advanced cupping (43%, 43%, 57%, and 57%, respectively) compared with mild cupping (0%) and moderate cupping (9%, 5%, 19%, and 33%, respectively). CONCLUSIONS: Eyes with GON and normal results of SAP have significantly greater structural and psychophysical abnormalities than do normal eyes evaluated by means of OCT, GDx VCC, SWAP, and FDT. Eyes with increased vertical cup-disc ratio are more likely to manifest such abnormalities on advanced diagnostic testing. PMID- 16476886 TI - Vitreous and aqueous penetration of orally administered moxifloxacin in humans. AB - OBJECTIVE: To investigate intraocular penetration of moxifloxacin hydrochloride after oral administration. METHODS: Prospective study of 15 patients scheduled for vitrectomy between September and November 2004 at the Barnes Retina Institute, St Louis, MO. Aqueous, vitreous, and serum samples were analyzed from 15 patients after oral administration of 2 tablets containing 400 mg of moxifloxacin. Assays were performed using high-performance liquid chromatography. RESULTS: The mean +/- SD moxifloxacin concentrations in plasma (n = 15), vitreous (n = 13), and aqueous (n = 13) samples were 3.56 +/- 1.31 microg/mL, 1.34 +/- 0.66 microg/mL, and 1.58 +/- 0.80 microg/mL, respectively. Mean +/- SD sampling times after oral administration of the second moxifloxacin tablet for plasma, vitreous, and aqueous were 2.94 +/- 0.81 hours, 3.77 +/- 0.92 hours, and 3.71 +/- 0.89 hours, respectively. The percentages of plasma moxifloxacin concentration in the vitreous and aqueous were 37.6% and 44.3%, respectively. Minimal inhibitory concentrations against 90% levels were exceeded against a wide spectrum of gram positive and gram-negative pathogens in the vitreous and aqueous. CONCLUSIONS: Moxifloxacin has a spectrum of coverage that encompasses the most common organisms in endophthalmitis. The pharmacokinetic findings of this investigation reveal that orally administered moxifloxacin achieves therapeutic levels in the noninflamed eye. Because of their broad spectrum of coverage, low minimal inhibitory concentration against 90% levels, good tolerability, and excellent oral bioavailability, fourth-generation fluoroquinolones may represent a major advance for managing posterior segment infections. PMID- 16476887 TI - Iris pigment epithelial translocation in the treatment of exudative macular degeneration: a 3-year follow-up. AB - OBJECTIVE: To report the functional and anatomical outcome of 20 patients who underwent surgical removal of choroidal neovascularization combined with transplantation of autologous iris pigment epithelial cells to the subretinal space 3 years after treatment. METHODS: Freshly isolated autologous iris pigment epithelial cells were translocated to the subretinal space in 20 patients after membrane extraction. Patients were followed up by funduscopy, angiography, microperimetry, and visual acuity testing. RESULTS: After a follow-up of 3 years, 1 patient showed improved visual acuity, 13 patients retained stable visual acuity, and 3 patients had reduced visual acuity. No macular edema or recurrent choroidal neovascularization was apparent at any time during the follow-up. CONCLUSIONS: Transplanted autologous iris pigment epithelial cells were well tolerated for 3 years and stabilization of visual acuity was achieved in most patients. These results suggest that iris pigment epithelial cells may serve as a substitute for retinal pigment epithelial cells after choroidal neovascularization removal in patients with exudative macular degeneration; however, whether these cells will be of any value for the restoration of vision and possible protection against choroidal neovascularization recurrence awaits further clinical observation and additional research. PMID- 16476888 TI - Normal macular thickness measurements in healthy eyes using Stratus optical coherence tomography. AB - OBJECTIVE: To report normal macular thickness measurements in healthy eyes using the latest commercially available optical coherence tomography (OCT) mapping software, version 3.0, from the Stratus OCT (OCT3). METHODS: Thirty-seven eyes from 37 healthy subjects underwent a complete ophthalmologic examination, including OCT. Six radial scans, 6 mm in length and centered on the fovea, were obtained using the OCT3. Retinal thickness was automatically calculated by OCT mapping software. Measurements were displayed as the mean and standard deviation for each of the 9 regions defined in the Early Treatment Diabetic Retinopathy Study. RESULTS: Foveal thickness (mean thickness in the central 1000-microm diameter area) and central foveal thickness (mean thickness at the point of intersection of 6 radial scans) on the OCT3 were 212 +/- 20 and 182 +/- 23 microm, respectively. Macular thickness measurements were thinnest at the center of the fovea, thickest within 3-mm diameter of the center, and diminished toward the periphery of the macula. The temporal quadrant was thinner than the nasal quadrant. Central foveal thickness was also manually determined as 170 +/- 18 microm, approximately 12 microm less than the value automatically obtained from the OCT3 software. There was no correlation between age and foveal thickness (P = .80). CONCLUSIONS: Mean foveal thickness measurements were 38 to 62 microm thicker than previously reported values, while mean central foveal thickness measurements were 20 to 49 microm thicker than previously published values. This discrepancy should be considered when interpreting OCT scans. PMID- 16476889 TI - Visual acuity at 10 years in Cryotherapy for Retinopathy of Prematurity (CRYO ROP) study eyes: effect of retinal residua of retinopathy of prematurity. AB - OBJECTIVE: To describe recognition (letter) acuity at age 10 years in eyes with and without retinal residua of retinopathy of prematurity (ROP). DESIGN: Presence and severity of ROP residua were documented by a study ophthalmologist. Masked testers measured monocular recognition visual acuity (Early Treatment of Diabetic Retinopathy Study) when the children were 10 years old. Two hundred forty-seven of 255 surviving Cryotherapy for Retinopathy of Prematurity (CRYO-ROP) randomized trial patients participated. A reference group of 102 of 104 Philadelphia-based CRYO-ROP study participants who did not develop ROP was also tested. RESULTS: More severe retinal residua were associated with worse visual acuity, regardless of whether retinal ablation was performed to treat the severe acute-phase ROP. However, within each ROP residua category, there was a wide range of visual acuity results. CONCLUSIONS: This is the first report of the relation between visual acuity (Early Treatment of Diabetic Retinopathy Study charts) and structural abnormalities related to ROP in a large group of eyes that developed threshold ROP in the perinatal period. Visual deficits are greater in eyes with more severe retinal residua than in eyes with mild or no residua. However, severity of ROP residua does not predict the visual acuity of an individual eye because within a single residua category, acuity may range from near normal to blind. PMID- 16476890 TI - Immunohistochemical evaluation of conjunctival fibrillin-1 in Marfan syndrome. AB - OBJECTIVE: To evaluate status of conjunctival fibrillin-1 in patients with Marfan syndrome with ectopia lentis. METHODS: Frozen sections of conjunctiva from 6 patients with Marfan syndrome with ectopia lentis and from 15 age-matched control subjects were stained with mouse antihuman fibrillin-1 antibody, using an avidin biotin immunoperoxidase technique. The fibrillin-1 staining characteristics of conjunctiva were analyzed with the light microscope. RESULTS: All the fresh frozen sections of conjunctival samples from control subjects demonstrated a characteristic pattern of fibrillin-1 staining. We observed a woven network of thin fibrils of uniform thickness surrounding collagen bundles. The fresh frozen samples from patients with Marfan syndrome showed consistent qualitative differences in fibrillin-1 staining when compared with samples from control subjects. The fibrils were longer and straighter than normal, varied in caliber, and showed fewer tendencies to form a woven pattern. CONCLUSIONS: Consistent, qualitative abnormalities in fibrillin-1 staining pattern can be seen in the conjunctiva of patients with Marfan syndrome with ectopia lentis. Conjunctival biopsy deserves further investigation as a diagnostic modality for Marfan syndrome in patients with ectopia lentis. PMID- 16476891 TI - Modified microkeratome-assisted posterior lamellar keratoplasty using a tissue adhesive. AB - OBJECTIVE: To compare graft stability and astigmatic change using suture vs tissue adhesive in an experimental model of microkeratome-assisted posterior lamellar keratoplasty. METHODS: A 300-microm-thick partial flap keratectomy was performed in human donor corneoscleral rims using an artificial anterior chamber and a manual microkeratome. The flap stopped at the left central opening border, providing a wide hinge to add stability. After flap reflection, a 6.25-mm trephination was performed to obtain a disc of posterior stroma, Descemet membrane, and endothelium. The disc was positioned in a sutureless fashion, and the flap secured with either 5 interrupted sutures or a chondroitin-sulfate aldehyde-based adhesive. Increasing intrachamber pressures were created to detect graft stability. Videokeratographic data were recorded to evaluate astigmatic change. RESULTS: The mean (SD) astigmatic change was 3.08 (0.84) diopters (D) in the sutured group and 1.13 (0.55) D in the glued group (P = .008). Mean (SD) resisted pressures were 95.68 (27.38) mm Hg and 82.45 (18.40) mm Hg in the sutured and glued groups, respectively (P = .97). CONCLUSION: This modified technique of microkeratome-assisted posterior lamellar keratoplasty showed excellent graft stability in both groups. Flaps sealed with the novel tissue adhesive had reduced astigmatic changes in our experimental model. CLINICAL RELEVANCE: Sutureless microkeratome-assisted posterior lamellar keratoplasty using tissue adhesive may become a new alternative in the surgical treatment of corneal endothelial disorders. PMID- 16476892 TI - Memantine protects neurons from shrinkage in the lateral geniculate nucleus in experimental glaucoma. AB - OBJECTIVE: To determine whether memantine as a treatment for glaucoma prevents neuron shrinkage in the lateral geniculate nucleus, the major target for retinal ganglion cells. METHODS: Sixteen monkeys with right-eye unilateral experimental glaucoma for 14 months were studied and treated with memantine (n = 9) or vehicle only (n = 7). Left lateral geniculate nucleus relay neurons (layers 1, 4, and 6) were examined following parvalbumin immunolabeling. Cell body cross-sectional areas and neuron numbers were assessed using unbiased methods. Memantine- and vehicle-treated glaucoma groups were compared using t tests and analysis of covariance. RESULTS: Compared with vehicle-treated animals, memantine-treated animals showed significantly less mean +/- SD neuron shrinkage in layers 1 (-4.0% +/- 13.9% vs 28.2% +/- 17.4%; P = .001) and 4 (24.9% +/- 10.0% vs 37.2% +/- 12.3%; P = .04). For layer 6, the difference was not statistically significant (34.2% +/- 10.1% vs 45.3% +/- 14.5%; P = .10). Analysis of covariance results showed significantly less neuron shrinkage in the memantine-treated group for layers 1, 4, and 6 (P < .001; P < .02; and P < .04, respectively). This difference was greatest in layer 1. In each of these layers, neuron numbers did not differ significantly between groups. CONCLUSION: Monkeys with glaucoma that were treated with memantine showed significantly less neuron shrinkage in the lateral geniculate nucleus than the vehicle-treated glaucoma group. CLINICAL RELEVANCE: The finding that memantine protects adult visual neurons from transsynaptic atrophy in experimental glaucoma could have therapeutic value. Currently, memantine is being tested in an ongoing clinical trial as a treatment for glaucoma. PMID- 16476893 TI - Quality of life after iodine 125 brachytherapy vs enucleation for choroidal melanoma: 5-year results from the Collaborative Ocular Melanoma Study: COMS QOLS Report No. 3. AB - OBJECTIVE: To describe health- and vision-targeted quality of life following treatment with iodine 125 brachytherapy vs enucleation for choroidal melanoma in a subgroup of patients who were treated and observed prospectively as part of a large randomized clinical trial. MAIN OUTCOME MEASURES: Difficulty with driving, near vision activities, and activities using stereopsis or binocularity; anxiety; and depression. PARTICIPANTS: Two hundred nine patients who enrolled in the Collaborative Ocular Melanoma Study trial for medium-sized tumors between March 1995 and July 1998 and gave informed consent prior to randomization to participation in an ancillary study of quality of life. METHODS: Patients were interviewed by telephone by a trained interviewer from the Collaborative Ocular Melanoma Study Coordinating Center at baseline (prior to randomization), at 6 months, and on annual anniversaries of enrollment. The questionnaire battery included the Medical Outcomes Study Short Form 36, the Activities of Daily Vision Scale, the National Eye Institute Visual Function Questionnaire, and the Hospital Anxiety and Depression Scale. Additional questions concerning satisfaction with posttreatment appearance and concerns about cancer recurrence also were included in posttreatment interviews. RESULTS: There was a significant increase in both treatment groups in levels of reported difficulty for most vision-oriented activities, and in bodily and ocular pain, 6 months following treatment. Differences in visual function between treatment groups reported during follow-up were relatively small, but significant differences favoring brachytherapy-treated patients were observed for driving during the first year of follow-up and for peripheral vision during the first 2 years of follow-up. Anxiety levels in both groups decreased significantly following treatment, but patients treated with brachytherapy with symptoms of anxiety were less likely to report later resolution of symptoms than patients with symptoms of anxiety who were treated with enucleation. This study was unable to assess impact of treatment on satisfaction with appearance and concern about cancer recurrence during the first year after treatment, but no treatment-related differences were found on these measures at 2 years and later follow-up times. CONCLUSIONS: Patients treated with brachytherapy reported significantly better visual function than patients treated with enucleation with respect to driving and peripheral vision for up to 2 years following treatment. Differences between treatments in visual function diminished by 3 to 5 years posttreatment, paralleling decline in visual acuity in brachytherapy-treated eyes. Patients treated with brachytherapy were more likely to have symptoms of anxiety during follow-up than patients treated with enucleation. APPLICATION TO CLINICAL PRACTICE: Given that no significant differences in survival between enucleation and brachytherapy have been found, the differences demonstrated here for driving and anxiety will allow the individual patient and physician to make informed choices regarding treatment based on personal preferences. PMID- 16476894 TI - Age-related eye disease, visual impairment, and survival: the Beaver Dam Eye Study. AB - OBJECTIVE: To investigate the relationship of age-related maculopathy, cataract, glaucoma, visual impairment, and diabetic retinopathy to survival during a 14 year period. METHODS: Persons ranging in age from 43 to 84 years in the period from September 15, 1987, to May 4, 1988, participated in the baseline examination of the population-based Beaver Dam Eye Study (n = 4926). Standardized protocols, including photography, were used to determine the presence of ocular disease. Survival was followed using standardized protocols. RESULTS: As of December 31, 2002, 32% of the baseline population had died (median follow-up, 13.2 years). After adjusting for age, sex, and systemic and lifestyle factors, poorer survival was associated with cortical cataract (hazard ratio [HR], 1.21; 95% confidence interval [CI], 1.06-1.37), any cataract (HR, 1.16; 95% CI, 1.03-1.32), diabetic retinopathy (HR per 1-step increase in 4-level severity, 1.36; 95% CI, 1.14 1.63), and visual impairment (HR, 1.24; 95% CI, 1.04-1.48) and marginally associated with increasing severity of nuclear sclerosis (HR, 1.07; 95% CI, 0.99 1.16). Age-related maculopathy and glaucoma were not associated with poorer survival. Associations tended to be slightly stronger in men than women. CONCLUSIONS: Cataract, diabetic retinopathy, and visual impairment were associated with poorer survival and not explained by traditional risk factors for mortality. These ocular conditions may serve as markers for mortality in the general population. PMID- 16476895 TI - Nine-year incidence of diabetic retinopathy in the Barbados Eye Studies. AB - OBJECTIVE: To estimate the 9-year incidence of diabetic retinopathy (DR) in a population with the same ancestry as African Americans. METHODS: Participants with diabetes mellitus and gradable photographs at the 9-year examination were evaluated (n = 436). The incidences of minimum/moderate/severe DR, clinically significant macular edema (CSME), and sight-threatening DR (severe DR plus CSME) were defined by the development of specific diabetic changes in persons without those conditions at baseline. Progression was defined as the development of severe/proliferative DR in persons with minimum/moderate DR at baseline. RESULTS: The 9-year DR incidence was 39.6% (38.0% for minimum, 9.0% for moderate, and 2.6% for severe/proliferative DR). Incidence tended to increase with diabetes duration and treatment. Of persons with preexisting DR at baseline, 8.2% progressed to proliferative DR. The CSME incidence was 8.7%, and it increased with diabetes duration, accounting for most of the overall incidence of sight-threatening DR. CONCLUSIONS: The study provides new data on long-term incidence among persons of African origin. Results suggest a possible lower risk of severe/proliferative DR than in whites, while CSME incidence seems comparable or higher. The main component of sight-threatening DR was CSME, highlighting the importance of DR as a cause of vision loss in this population. PMID- 16476896 TI - Association of adipose and red blood cell lipids with severity of dominant Stargardt macular dystrophy (STGD3) secondary to an ELOVL4 mutation. AB - OBJECTIVE: To determine whether adipose and red blood cell membrane lipids, particularly long-chain polyunsaturated fatty acids such as docosahexaenoic acid and eicosapentaenoic acid, are significantly correlated with phenotype in a family with autosomal dominant Stargardt macular dystrophy (gene locus STGD3). A mutation in the ELOVL4 gene is responsible for the macular dystrophy in this family, and its disease-causing mechanism may be its possible involvement in fatty acid elongation in the retina. METHODS: The subjects in this study included 18 adult family members known to have a 2-base pair deletion in the ELOVL4 gene. Control subjects included 26 family members without the mutation. Each subject received a complete eye examination including fundus photographs, the results of which were used to grade the severity of macular dystrophy on a 3-tier scale. Red blood cell membrane and adipose tissue lipids were analyzed as an indication of short-term and long-term dietary fatty acid intake. RESULTS: When adipose lipids were analyzed, there was a significant inverse relationship between phenotypic severity and the level of eicosapentaenoic acid (r = -0.54; P = .04). When red blood cell lipids were analyzed, there were significant inverse relationships between phenotypic severity and levels of eicosapentaenoic acid (r = -0.55; P = .02) and docosahexaenoic acid (r = -0.48; P = .04). CONCLUSIONS: These results indicate that the phenotypic diversity in this family may be related to differences in dietary fat intake as reflected by adipose and red blood cell lipids. CLINICAL RELEVANCE: This study demonstrates that dietary factors can influence the severity of an inherited human macular dystrophy. PMID- 16476897 TI - Resident physician mentoring program in ophthalmology: the Tennessee experience. AB - OBJECTIVE: To establish a mentoring program to provide resident physicians in ophthalmology with career guidance in practice management and to identify new and creative ways to involve future eye physicians in the legislative and political process. METHODS: A multicenter prospective study was conducted of the mentorship experiences of 24 (88.9%) of 27 resident physicians in Tennessee during the 2000 2001 academic year. Participants were assigned into 1 of 3 groups: an active mentorship group, a passive mentorship group, and a no mentorship group. The active mentorship group participated in preceptorship activities with "mentor" community-based eye physicians and scheduled meetings with state legislators and regulators. The active mentorship and passive mentorship groups attended a 1-day practice management seminar, but the no mentorship group received no formal mentorship during the 4-month study period. A survey instrument was given to all participants before and after the 4-month study period. RESULTS: Following completion of the mentorship program, the active mentorship group had favorable changes in perceptions and attitudes toward medical organizations (P<.03) when compared with baseline prementorship responses. Compared with the no mentorship group, the active mentorship group also reported an increased willingness to make political campaign donations (P<.05) and expressed an increased desire for the Tennessee Academy of Ophthalmology to offer practice management programs (P<.02). CONCLUSION: A short 4-month mentorship program can elicit favorable changes in residents' perceptions and attitudes toward medical professional organizations. Additional opportunities may lie with a lengthier and more intensive mentoring program. PMID- 16476898 TI - Nocturnal rhythms of intraocular pressure. PMID- 16476899 TI - Congenital ectropion uveae and glaucoma. PMID- 16476900 TI - Melanocytoma of the optic nerve associated with sound-induced phosphenes. PMID- 16476902 TI - Vitreopapillary traction confirmed by optical coherence tomography. PMID- 16476901 TI - Ocular adverse effects associated with cyclooxygenase-2 inhibitors. PMID- 16476903 TI - Ultra-high resolution optical coherence tomography of retinal pigment epithelial tear following blunt trauma. PMID- 16476904 TI - Spinal fluid leak after chiropractic manipulation of the cervical spine. PMID- 16476905 TI - Orbital foreign body and ruptured globe from needlefish impalement. PMID- 16476906 TI - Contradictions in the amblyopia treatment studies. PMID- 16476907 TI - Optical coherence tomography findings in macular hole due to argon laser burn. PMID- 16476908 TI - Home exercises for convergence insufficiency in children. PMID- 16476909 TI - Alternative approach to the treatment of subepithelial fibrosis following radial keratotomy. PMID- 16476910 TI - Thoracic outlet syndrome in a competitive baseball player secondary to the anomalous insertion of an atrophic pectoralis minor muscle: a case report. PMID- 16476911 TI - The anterior cruciate ligament tear rate varies by race in professional Women's basketball. AB - BACKGROUND: Female basketball players are more likely to tear their anterior cruciate ligament than are their male counterparts. Many causes are postulated for the difference observed in the rate of anterior cruciate ligament tears between genders. However, little is known about the differences in tears within gender. HYPOTHESIS: The rate of anterior cruciate ligament tears is different in White European American female basketball players and non-White European American players. STUDY DESIGN: Cohort study (Prevalence); Level of evidence, 2. METHODS: The authors investigated the differences in anterior cruciate ligament tears by risk exposure in women of different racial or ethnic backgrounds playing in the Women's National Basketball Association for the 1999 through 2003 seasons. Using the injury surveillance data from the Women's National Basketball Association, the authors compared anterior cruciate ligament injuries in White European American and non-White European American players to determine differences in the rate of anterior cruciate ligament tears between racial groups. RESULTS: The anterior cruciate ligament tear rate for White European American players was 0.45 per 1000 athletic exposures, whereas for non-White European American players (black or African American, Hispanic, and Asian players) the rate was 0.07. The odds ratio of anterior cruciate ligament tears in White European American versus non-White European American players was 6.55 (95% confidence interval, 1.35 31.73). CONCLUSION: Our retrospective study shows that the anterior cruciate ligament tear rate for White European American players was 0.45 per 1000 athletic exposures, whereas tear rates in the Women's National Basketball Association vary by racial group, with White European American players having more than 6 times the anterior cruciate ligament tear rate of other ethnic groups combined. Further prospective studies of athletes are needed to validate this finding and shed light on possible reasons. PMID- 16476912 TI - Recalcitrant patellar tendinosis in elite athletes: surgical treatment in conjunction with aggressive postoperative rehabilitation. AB - BACKGROUND: Recalcitrant patellar tendinosis is difficult to treat, and results are varied. HYPOTHESIS: Surgical removal of necrotic tissue, surgical stimulation of remaining tendon, and aggressive and specific rehabilitation after patellar tendonectomy will allow athletes to return to sports. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: From December 1996 to July 2002, 16 high-level athletes (4 professional, 2 Olympic, 9 collegiate, 1 preparatory), aged 16 to 25 years (mean, 19.7 years), with 22 symptomatic patellar tendons had failed nonoperative care of their patellar tendinosis symptoms and were unable to compete effectively in their sports. Magnetic resonance imaging showed confirmation of disease, with typical findings being necrosis in the posterior half of an abnormally thick patellar tendon, often in conjunction with partial tearing of the posterior half with a compensatory enlargement of the anterior half. Each patient then underwent tendonectomy of the necrotic portion in conjunction with stimulation of the remaining tendon by making multiple longitudinal cuts in the tendon. Patients participated in a postoperative rehabilitation protocol that included immediate range of motion, full flexion, and immediate high-repetition, low-resistance quadriceps muscle exercise. RESULTS: Subjective improvement was noted in all athletes. Return to the same sport at prior level of intensity was accomplished by 14 of 16 patients (87.5%) at a mean of 8.1 months (range, 3-12 months). CONCLUSION: Overall, tendonectomy, surgical tendon stimulation, and aggressive postoperative rehabilitation were found to be a safe, effective way to return high-level athletes to their sports. PMID- 16476913 TI - Effects of stretching on passive muscle tension and response to eccentric exercise. AB - BACKGROUND: Stretching is used in an attempt to improve performance and reduce the risk of muscle injury, with little evidence to support its effectiveness. HYPOTHESIS: Four weeks of static or ballistic stretching can attenuate the increased soreness and decreased flexibility seen after eccentric exercise. STUDY DESIGN: Controlled laboratory study. METHODS: Twenty-nine male subjects were randomly assigned to a static stretching, ballistic stretching, or control group. On each of 4 consecutive days, they completed 4 maximal range of motion stretches using a Cybex isokinetic dynamometer to passively stretch the hamstrings at 0.087 rad.s(-1) (5 deg.s(-1)). Stiffness from 0.87 to 1.48 rad (50 degrees -85 degrees ), peak range of motion, work absorption, peak resistive torque, and soreness were measured. Participants then completed 4 weeks of either static or ballistic stretching for a total stretching duration of 3600 seconds. After training, the 4 days of testing were repeated with an eccentric exercise task added after day 1. RESULTS: Stretching groups had an increase in range of motion and stretch tolerance after 4 weeks of stretching, with no change in muscle stiffness, work absorption, or delayed onset muscle soreness. After eccentric exercise, they also had greater range of motion and stretch tolerance than did controls. CONCLUSION: Both static stretching and ballistic stretching increase range of motion, most likely as a result of enhanced stretch tolerance rather than changes in muscle elasticity. Four weeks of stretching maintain range of motion and stretch tolerance in the days after eccentric exercise. PMID- 16476914 TI - Prospective evaluation of allograft meniscus transplantation: a minimum 2-year follow-up. AB - BACKGROUND: Clinical and biomechanical studies have demonstrated the increase in contact pressure and progressive deterioration of the tibiofemoral compartments that occur after partial or complete meniscectomy. Meniscus transplantation has been indicated for the symptomatic postmeniscectomy patient to alleviate symptoms and potentially prevent the progression of articular degeneration. PURPOSE: To report the early-term results after allograft meniscus transplantations from a single institution performed by a single surgeon. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Forty-four meniscus transplants in 39 patients were evaluated at minimum 2-year follow-up using the Lysholm, Tegner, International Knee Documentation Committee, Knee Injury and Osteoarthritis Outcome Score, Noyes symptom rating and sports activity, and SF-12 scoring systems; visual analog pain scales; patient satisfaction; and physical examination. Four transplants failed early, leaving 40 transplants in 36 patients for review. Patients were grouped into medial and lateral transplant groups as well as those with isolated and combined procedures. Twenty-one menisci were transplanted in isolation (52.5%), and 19 were combined with other procedures (47.5%) to address concomitant articular cartilage injury. RESULTS: Patients demonstrated statistically significant improvements in standardized outcomes surveys and visual analog pain and satisfaction scales. In 7 patients, treatment had failed at final follow-up. Overall, 77.5% of patients reported they were completely or mostly satisfied with the procedure, and 90% of patients were classified as normal or nearly normal using the International Knee Documentation Committee knee examination score at final follow-up. There were no significant differences in the medial and lateral subgroups, although the lateral subgroup did demonstrate a trend toward greater improvement. No significant differences were noted in the isolated and combined subgroups. CONCLUSION: Meniscus transplantation alone or in combination with other reconstructive procedures results in reliable improvements in knee pain and function at minimum 2-year follow-up. Longer term studies are necessary to determine if transplantation can prevent the articular degeneration associated with meniscectomy. PMID- 16476915 TI - The effect of a balance training program on the risk of ankle sprains in high school athletes. AB - BACKGROUND: Ankle sprains are the most common musculoskeletal injuries that occur in athletes, and they have a profound impact on health care costs and resources. HYPOTHESIS: A balance training program can reduce the risk of ankle sprains in high school athletes. STUDY DESIGN: Randomized controlled clinical trial; Level of evidence, 1. METHODS: Seven hundred and sixty-five high school soccer and basketball players (523 girls and 242 boys) were randomly assigned to either an intervention group (27 teams, 373 subjects) that participated in a balance training program or to a control group (28 teams, 392 subjects) that performed only standard conditioning exercises. On-site athletic trainers recorded athlete exposures and sprains. RESULTS: The rate of ankle sprains was significantly lower for subjects in the intervention group (6.1%, 1.13 of 1000 exposures vs 9.9%, 1.87 of 1000 exposures; P = .04). Athletes with a history of an ankle sprain had a 2-fold increased risk of sustaining a sprain (risk ratio, 2.14), whereas athletes who performed the intervention program decreased their risk of a sprain by one half (risk ratio, 0.56). The ankle sprain rate for athletes without previous sprains was 4.3% in the intervention group and 7.7% in the control group, but this difference was not significant (P = .059). CONCLUSION: A balance training program will significantly reduce the risk of ankle sprains in high school soccer and basketball players. PMID- 16476916 TI - Double-loop suture repair for acute acromioclavicular joint disruption. AB - BACKGROUND: Although it has been established that surgical treatment for acromioclavicular joint disruption (types IV-VI and type III in overhead throwing athletes and heavy laborers) is preferred, the literature is inconclusive about the best type of surgery. PURPOSE: With the goal of avoiding the potential complications of hardware use, the authors present a coracoclavicular functional stabilization technique with the intention to restore the anteroposterior and superior displacement of the clavicle. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: From 1999 to 2003, 38 patients with an acute, complete acromioclavicular joint separation (34 men, 4 women; mean age, 33.5 years) underwent surgical reconstruction with the described coracoclavicular loop stabilization technique. With this technique, the superior and anteroposterior displacement of the clavicle can be easily controlled using 2 pairs of Ethibond No. 5 nonabsorbable sutures-one passed in front and the other behind the clavicle, through a central drill hole, 2 cm from its lateral end, directly above the base of the coracoid process (at the corresponded attachment of coracoclavicular ligaments). Passive shoulder motion was encouraged by the second postoperative day. RESULTS: Thirty-four patients were available for the last clinical and radiologic evaluation. At a mean follow-up of 33.2 months (range, 18 59 months), the mean Constant-Murley score was 93.5 points (range, 73-100 points), and 2 cases with slight loss of reduction (less than half of the width of the clavicle) were detected. Complications included 1 case with superficial infection and 1 patient (basketball player) with persistent tenderness in the acromioclavicular joint without signs of secondary arthritis. The incidence of periarticular ossification was 17.6% and did not affect the final outcome. Secondary degenerative changes were not detected. CONCLUSION: Considering the nearly anatomical reconstruction, the avoidance of hardware complications, and the low rate of recurrence, this technique may be an attractive alternative to the management of acute acromioclavicular joint separations. PMID- 16476917 TI - Weakness in end-range plantar flexion after Achilles tendon repair. AB - BACKGROUND: Separation of tendon ends after Achilles tendon repair may affect the tendon repair process and lead to postoperative end-range plantarflexion weakness. HYPOTHESIS: Patients will have disproportionate end-range plantarflexion weakness after Achilles tendon repair. STUDY DESIGN: Descriptive laboratory study. METHODS: Four-strand core suture repairs of Achilles tendon were performed on 1 female and 19 male patients. Postoperatively, patients were nonweightbearing with the ankle immobilized for 4 weeks. Plantarflexion torque, dorsiflexion range of motion, passive joint stiffness, toe walking, and standing single-legged heel rise (on an incline, decline, and level surface) were assessed after surgery (mean, 1.8 years postoperative; range, 6 months-9 years). Maximum isometric plantarflexion torque was measured at 20 degrees and 10 degrees of dorsiflexion, neutral, and 10 degrees and 20 degrees of plantar flexion. Percentage strength deficit (relative to noninvolved leg) was computed at each angle. Passive dorsiflexion range of motion was measured goniometrically. Passive joint stiffness was computed from increase in passive torque between 10 degrees and 20 degrees of dorsiflexion, before isometric contractions. RESULTS: Significant plantarflexion weakness was evident on the involved side at 20 degrees and 10 degrees of plantar flexion (34% and 20% deficits, respectively; P <.001), with no torque deficits evident at other angles (6% at neutral, 3% at 10 degrees of dorsiflexion, 0% at 20 degrees of dorsiflexion). Dorsiflexion range of motion was not different between involved and noninvolved sides (P = .7). Passive joint stiffness was 34% lower on the involved side (P <.01). All patients could perform an incline heel rise; 14 patients could not perform a decline heel rise (P <.01). CONCLUSION: Disproportionate weakness in end-range plantar flexion, decreased passive stiffness in dorsiflexion, and inability to perform a decline heel rise are evident after Achilles tendon repair. Possible causes include anatomical lengthening, increased tendon compliance, and insufficient rehabilitation after Achilles tendon repair. CLINICAL RELEVANCE: Impairments will have functional implications for activities (eg, descending stairs and landing from a jump). Weakness in end-range plantar flexion may be an unrecognized problem after Achilles tendon repair. PMID- 16476919 TI - A comparison between clinical assessment and magnetic resonance imaging of acute hamstring injuries. AB - BACKGROUND: Physicians evaluating hamstring strains in professional football players are increasingly turning to magnetic resonance imaging to support the clinical diagnosis and management of the injury. However, little information is available to assess how magnetic resonance imaging compares with the clinical evaluation in establishing the duration of rehabilitation required. HYPOTHESIS: Magnetic resonance imaging of hamstring strains can be useful in determining duration of rehabilitation. STUDY DESIGN: Cohort study (Diagnosis); Level of evidence, 1. METHODS: Fifty-eight professional football players with a diagnosis of hamstring injury made by the team physician were enrolled in the study. All players underwent magnetic resonance imaging and a clinical evaluation by an independent physical therapist within 3 days of the injury. Presence, type, and location of injury were recorded in each examination. The physical therapist estimated the time required until return to competition, and the radiologist used the length of the injury (coronal view) to establish rehabilitation duration. Both clinicians were blinded to the other modality. RESULTS: Clinical and magnetic resonance imaging assessments were in agreement in 38 of 58 cases (65.5%). In 18 cases (31.0%), a clinically positive diagnosis was made, but no abnormalities were evident on magnetic resonance imaging. In 2 cases (3.4%), magnetic resonance imaging detected an injury, whereas the clinical examination had negative or equivocal findings. Both clinical examination and magnetic resonance imaging findings were strongly correlated with the actual time required to return to competition (r = .69, P < .001 and r = .58, P < .001, respectively). The correlation coefficient between clinical predictions and magnetic resonance imaging findings was moderate (r = .36, P = .006). CONCLUSION: This study shows that magnetic resonance imaging is not required for estimating the duration of rehabilitation of an acute minor or moderate hamstring injury in professional football players. PMID- 16476918 TI - Distal biceps tendon repair: a biomechanical comparison of intact tendon and 2 repair techniques. AB - BACKGROUND: A variety of techniques have been described for distal biceps tendon reattachment-bone tunnel with transosseous sutures, suture anchors, and interference screw techniques. HYPOTHESIS: There will be no significant difference between the mean failure strength, maximum strength, and stiffness of the intact specimen and repair techniques tested: bone tunnel with transosseous sutures and interference screw. STUDY DESIGN: Controlled laboratory study. METHODS: Nine matched pairs of fresh-frozen human cadaveric elbows were prepared. The intact tendon was pulled from the radial tuberosity; the right and left elbows were randomized to bone tunnels with transosseous sutures or interference screw repair techniques. The repaired specimens were pulled using the same regimen for the intact tendon. Failure strength, maximum strength, and stiffness were measured and compared. RESULTS: The mean failure strength, maximum strength, and stiffness of intact tendons were 204.3 +/- 76.9 N, 221.7 +/- 65.9 N, and 30.1 +/- 12.4 N/mm, respectively; for the interference screw specimens, 178.0 +/- 54.5 N, 192.1 +/- 53.1 N, and 30.4 +/- 9.5 N/mm, respectively; and for the bone tunnel specimens, 124.9 +/- 22.8 N, 206.6 +/- 49.8 N, and 15.9 +/- 5.6 N/mm, respectively. There were no significant differences between measures in the intact and interference screw specimens. Mean failure strength and stiffness of the bone tunnel specimens were significantly lower than those of the intact and interference screw specimens; there was no significant difference between the maximum strengths of the treatments. Interference screw failure occurred abruptly with little plastic deformation in nearly all specimens with the tendon and screw pulling out as a unit, often involving fracture of the radial wall. Two of the bone tunnels failed at the bony bridge; the remainder lost bone-tendon contact as the distal tendon was shredded by the suture. CONCLUSION: The results suggest interference screw fixation repair is nearly as strong and stiff as the intact tendon and stronger than the bone tunnel repair technique. CLINICAL RELEVANCE: The interference screw provides better stiffness and failure strength compared with the bone tunnel technique for distal biceps tendon repair. Given the superior mechanical properties, the interference screw technique is recommended as the treatment of choice for biceps tendon rupture repair. PMID- 16476920 TI - The COX-2 specific inhibitor Valdecoxib versus tramadol in acute ankle sprain: a multicenter randomized, controlled trial. AB - BACKGROUND: The cyclooxygenase-2 specific inhibitor valdecoxib has not been approved in the United States for treatment of acute pain. HYPOTHESIS: Valdecoxib 20 mg twice daily or once daily (both with a 40-mg loading dose) is not clinically inferior to tramadol for treating the signs and symptoms of acute ankle pain. STUDY DESIGN: Randomized, controlled clinical trial; Level of evidence, 1. METHODS: Patients (N = 829) with acute first- or second-degree ankle sprain received 7 days' treatment with valdecoxib 20 mg either twice daily or once daily (both with 40-mg loading dose), tramadol 50 mg 4 times daily, or placebo. The primary end point was Patient's Assessment of Ankle Pain visual analog scale on day 4; a test of noninferiority compared valdecoxib with tramadol. RESULTS: On day 4, both valdecoxib doses were significantly better versus placebo and were comparable with tramadol in relieving ankle pain. On day 7, valdecoxib, but not tramadol, significantly reduced pain versus placebo. On days 4 and 7, more patients resumed normal walking with valdecoxib (45%-47% and 73%-79%, respectively) than with placebo (35% and 64%, respectively) or tramadol (38% and 67%, respectively). In contrast to valdecoxib, the number of withdrawals due to adverse events was significantly higher in the tramadol group (12.2% vs 3.4%; P = .0005). CONCLUSIONS: Valdecoxib was comparable with tramadol and was significantly better than placebo in treating acute ankle sprain, and it enabled more patients to resume normal walking on days 4 and 7. Both valdecoxib and tramadol were well tolerated. PMID- 16476922 TI - Hemi tics and deep brain stimulation. PMID- 16476921 TI - Low-intensity pulsed ultrasound accelerates and a nonsteroidal anti-inflammatory drug delays knee ligament healing. AB - BACKGROUND: Low-intensity pulsed ultrasound and nonsteroidal anti-inflammatory drugs are used to treat ligament injuries; however, their individual and combined effects are not established. HYPOTHESES: Low-intensity pulsed ultrasound accelerates ligament healing, a nonsteroidal anti-inflammatory drug delays healing, and the nonsteroidal anti-inflammatory drug inhibits the beneficial effect of low-intensity pulsed ultrasound. STUDY DESIGN: Controlled laboratory study. METHODS: Sixty adult rats underwent bilateral transection of their knee medial collateral ligaments. Animals were divided into 2 drug groups and treated 5 d/wk with celecoxib (5 mg/kg) mixed in a vehicle solution (NSAID group) or vehicle alone (VEH group). One to 3 hours after drug administration, all animals were treated with unilateral active low-intensity pulsed ultrasound and contralateral inactive low-intensity pulsed ultrasound. Equal numbers of animals from each drug group were mechanically tested at 2 weeks (n = 14/group), 4 weeks (n = 8/group), and 12 weeks (n = 8/group) after injury. RESULTS: Ultrasound and drug intervention did not interact to influence ligament mechanical properties at any time point. After 2 weeks of intervention, ligaments treated with active low intensity pulsed ultrasound were 34.2% stronger, 27.0% stiffer, and could absorb 54.4% more energy before failure than could ligaments treated with inactive low intensity pulsed ultrasound, whereas ligaments from the NSAID group could absorb 33.3% less energy than could ligaments from the VEH group. There were no ultrasound or drug effects after 4 and 12 weeks of intervention. CONCLUSIONS: Low intensity pulsed ultrasound accelerated but did not improve ligament healing, whereas the nonsteroidal anti-inflammatory drug delayed but did not impair healing. When used in combination, the beneficial low-intensity pulsed ultrasound effect was cancelled by the detrimental nonsteroidal anti-inflammatory drug effect. CLINICAL RELEVANCE: Low-intensity pulsed ultrasound after ligament injury may facilitate earlier return to activity, whereas non-steroidal anti inflammatory drugs may elevate early reinjury risk. PMID- 16476923 TI - Extensive brain and muscular cysticercosis. PMID- 16476924 TI - What are we seeing? Is posterior cortical atrophy just Alzheimer disease? PMID- 16476925 TI - Trial of dichloroacetate in MELAS: toxicity overshadows the assessment of potential benefit. PMID- 16476926 TI - Is myelination the precipitating neural event for language development in infants and toddlers? PMID- 16476927 TI - IV tissue plasminogen activator use in acute stroke: experience from a statewide registry. AB - OBJECTIVE: To assess the use of IV recombinant tissue plasminogen activator (rt PA) in a statewide hospital-based stroke registry and to identify factors associated with its use among eligible patients. METHODS: A modified stratified sampling scheme was used to obtain a representative sample of 16 hospitals. Prospective case ascertainment and data collection were used to identify all acute stroke admissions over a 6-month period. Subjects eligible for IV rt-PA were defined as those who arrived within 3 hours of onset, who had no evidence of hemorrhage on initial brain image, and who had no physician-documented reasons for non-treatment with IV rt-PA. Multivariate logistic regression was used to identify factors associated with IV rt-PA use. RESULTS: Of 2,566 stroke admissions, 330 (12.9%) met the eligibility criteria for rt-PA treatment, and of these 43 (13%) received IV rt-PA treatment. Among 2,236 admissions excluded from consideration, 21% had evidence of hemorrhage on initial imaging, 35% had unknown stroke onset times, 38% had an onset to arrival time >3 hours, and 6% had physician documented contraindications. Among eligible patients, being male, use of emergency medical services, and rapid presentation were associated with increased IV rt-PA use. CONCLUSIONS: Treatment with IV rt-PA was underutilized in this hospital-based stroke registry. The primary reason for nontreatment was delayed presentation. Reducing prehospital and in-hospital response times would help increase IV rt-PA use, as would greater emergency medical services use. Improving the documentation of onset times would help clarify the underlying causes of delayed presentation. PMID- 16476928 TI - Secondary-prevention drug prescription in the very elderly after ischemic stroke or TIA. AB - BACKGROUND: An age bias may exist in the prescription of important secondary preventive therapies in the elderly. OBJECTIVE: To evaluate patterns of drug prescription for cardiovascular prevention in the very elderly following hospitalization for an acute ischemic stroke or TIA. METHODS: The authors compared subjects ages > or = 80 with those < 80 in the California Acute Stroke Prototype Registry to evaluate the impact of age on receipt of secondary prevention medications at the time of hospital discharge. Prespecified secondary prevention drug classes studied were antithrombotics, lipid-lowering agents, and antihypertensives. RESULTS: Overall, there were 260 patients age > or = 80 and 534 age < 80 admitted with stroke or TIA during the study period. Patients > or = 80 years were less likely to receive actual treatment with antithrombotic medications (p = 0.002) and lipid-lowering medications (p = 0.005) but were more likely to receive antihypertensive medications (p = 0.0007) than their younger counterparts. With regard to optimal treatment (defined as receipt of, or a valid contraindication to, treatment in each category), those > or = 80 were equally likely to receive antithrombotic medications and lipid therapy but remained more likely to receive antihypertensive treatment (77.7 vs 67.0%; p = 0.0007). There were no differences in receipt of optimal combination therapy (defined as optimal treatment in all three therapeutic classes) between patient age groups, even when adjusted for medical history. CONCLUSION: After hospitalization for stroke or TIA, no differences in overall optimal treatment prescription of secondary prevention medications between patients ages > or = 80 and their younger counterparts were observed. PMID- 16476929 TI - Dichloroacetate causes toxic neuropathy in MELAS: a randomized, controlled clinical trial. AB - OBJECTIVE: To evaluate the efficacy of dichloroacetate (DCA) in the treatment of mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS). BACKGROUND: High levels of ventricular lactate, the brain spectroscopic signature of MELAS, correlate with more severe neurologic impairment. The authors hypothesized that chronic cerebral lactic acidosis exacerbates neuronal injury in MELAS and therefore, investigated DCA, a potent lactate-lowering agent, as potential treatment for MELAS. METHODS: The authors conducted a double-blind, placebo-controlled, randomized, 3-year cross-over trial of DCA (25 mg/kg/day) in 30 patients (aged 10 to 60 years) with MELAS and the A3243G mutation. Primary outcome measure was a Global Assessment of Treatment Efficacy (GATE) score based on a health-related event inventory, and on neurologic, neuropsychological, and daily living functioning. Biologic outcome measures included venous, CSF, and 1H MRSI-estimated brain lactate. Blood tests and nerve conduction studies were performed to monitor safety. RESULTS: During the initial 24-month treatment period, 15 of 15 patients randomized to DCA were taken off study medication, compared to 4 of 15 patients randomized to placebo. Study medication was discontinued in 17 of 19 patients because of onset or worsening of peripheral neuropathy. The clinical trial was terminated early because of peripheral nerve toxicity. The mean GATE score was not significantly different between treatment arms. CONCLUSION: DCA at 25 mg/kg/day is associated with peripheral nerve toxicity resulting in a high rate of medication discontinuation and early study termination. Under these experimental conditions, the authors were unable to detect any beneficial effect. The findings show that DCA-associated neuropathy overshadows the assessment of any potential benefit in MELAS. PMID- 16476930 TI - The cognitive profile of posterior cortical atrophy. AB - BACKGROUND: Posterior cortical atrophy (PCA) is a progressive dementia characterized by prominent disorders of higher visual processing, affecting both dorsal and ventral streams to cause Balint's syndrome, alexia, and visual agnosia. OBJECTIVE: To define the cognitive profile of PCA and compare to the typical, primary amnestic dementia of the Alzheimer's type (DAT). METHODS: The authors used standard cognitive tests and a novel battery designed to reflect dysfunction in both ventral (Object, Face & Color Agnosia Screen [OFCAS]) and dorsal (complex pictures and compound stimuli) visual streams. The authors identified 19 patients with PCA and compared their performance to a matched group of patients with DAT and normal controls. RESULTS: Patients with PCA were younger with marked impairment in visuospatial tasks, reading, and writing but relative preservation of memory compared to DAT using standard tests. Dorsal stream signs were most prevalent among the patients with PCA with no pure ventral stream syndromes found. All novel tests distinguished reliably between subjects with complex picture descriptions and processing of compound stimuli showing the most significant differences compared to DAT. CONCLUSIONS: PCA is predominantly a dorsal stream syndrome, distinct from typical DAT, which involves occipitotemporal regions over time. PMID- 16476931 TI - Myelination of language-related areas in the developing brain. AB - BACKGROUND: The rapid development of language abilities in early childhood coincides with a similarly accelerated progression in brain maturation. OBJECTIVE: To quantitate myelination in the lateral part of the verbal left hemisphere from birth to 3 years in the living human brain. METHODS: One hundred children (mean age 16.6 months) were examined using three-dimensional MRI, and a subgroup of 40 children were also evaluated behaviorally. The volume of myelinated white matter was measured in language-related temporal and frontal regions and in the central sensorimotor region. A method was developed to compose a movie sequence for all the myelination process using volumetric data. RESULTS: A plot of age against relative volume of myelinated white matter graphically detailed the myelination progress in the lateral brain. The changes started in sensorimotor white matter and the Heschl gyrus and ultimately extended to the language-related areas. Both comprehension and production regions showed a very similar myelination course, suggesting simultaneous maturation of the temporofrontal language network. The movie sequence of white matter images dynamically displayed the anatomic details of myelin deposition in this part of the brain. The analysis of language performance showed acceleration in children's vocabulary after 18 months, once a rapid myelination phase was attained in the language brain. CONCLUSIONS: This volumetric study may contribute to further characterize the early stages of brain maturation by showing the fine progression of myelin deposition in the language domains and illustrating its relationship to children's vocabulary acquisition. PMID- 16476932 TI - Familial risk of migraine: variation by proband age at onset and headache severity. AB - BACKGROUND: It is well established that migraine aggregates within families. Less is known about the influence of proband characteristics (e.g., age at onset, headache severity) on familial aggregation. OBJECTIVE: To examine the association between the proband's migraine severity and age at migraine onset and familial aggregation of migraine. METHODS: The authors investigated the migraine prevalence in first-degree relatives of 532 persons with migraine and control subjects in a population study. Familial aggregation was expressed as the risk of migraine in family members of probands divided by risk in control family members. RESULTS: The relative risk (RR) of migraine in first-degree relatives of migraine probands was elevated compared with family members of controls (RR = 1.88; 95% CI: 1.30 to 2.72). The RR was also significantly higher for relatives of probands reporting onset of migraine before age 16 (2.50; 95% CI: 1.65 to 3.79) compared with those with onset at age 16 or older (1.44; 95% CI: 0.93 to 2.23). Among probands with very severe average pain scores (i.e., 9 to 10 on a 0 to 10 scale), the RR of migraine in family members was 2.38 (95% CI: 1.56 to 3.62) compared with 1.52 (0.99 to 2.34) for less severe pain (p < 0.05). CONCLUSION: Early onset of migraine in the proband as well as the severity of migraines are associated with higher levels of family aggregation. PMID- 16476933 TI - Oral contraceptives and increased headache prevalence: the Head-HUNT Study. AB - OBJECTIVE: To examine the prevalence of headache and migraine among women using oral contraceptives (OCs) in a large, cross-sectional population-based study. METHODS: In the Nord-Trondelag Health Study in Norway 1995-1997 (HUNT 2), 27,700 (60%) out of 46,506 invited women responded to headache questions (Head-HUNT). Among 14,353 premenopausal women, 13,944 (97%) responded to questions regarding use of contraceptives. RESULTS: There was a significant association between headache and reported use of estrogen-containing OCs in premenopausal women, both for migraine (OR = 1.4, 95% CI = 1.2 to 1.7) and for non-migrainous headache (OR = 1.2, 95% CI = 1.0 to 1.4). A significant dose relationship between headache and the amount of estrogen in the OCs could not be demonstrated. No significant association between headache and OCs containing only gestagen was found. CONCLUSION: Headache, especially migraine, was more likely among premenopausal women using oral contraceptives containing estrogen. PMID- 16476934 TI - Tetrabenazine as antichorea therapy in Huntington disease: a randomized controlled trial. AB - BACKGROUND: Tetrabenazine (TBZ) selectively depletes central monoamines by reversibly binding to the type 2 vesicular monoamine transporter. Open-label reports indicate TBZ is effective in treating chorea. OBJECTIVE: To examine the safety, efficacy, and dose tolerability of TBZ for treating chorea in Huntington disease (HD). METHODS: The authors randomized 84 ambulatory patients with HD to receive TBZ (n = 54) or placebo (n = 30) for 12 weeks. TBZ was increased over 7 weeks up to a maximum of 100 mg/day or until the desired antichoreic effect occurred or intolerable adverse effects supervened. The primary outcome was the change from baseline in the chorea score of the Unified Huntington's Disease Rating Scale (UHDRS) RESULTS: TBZ treatment resulted in a reduction of 5.0 units in chorea severity compared with a reduction of 1.5 units on placebo treatment (adjusted mean effect size = -3.5 +/- 0.8 UHDRS units [mean +/- SE]; 95% CI: 5.2, -1.9; p < 0.0001). There was also a significant benefit on ratings of clinical global improvement. There were five study withdrawals in the TBZ group and five serious adverse events (SAEs) in four subjects (drowning suicide, complicated fall, restlessness/suicidal ideation, and breast cancer) compared with one withdrawal and no SAEs in the placebo group. CONCLUSION: Tetrabenazine (TBZ), at adjusted dosages of up to 100 mg/day, effectively lessens chorea in ambulatory patients with Huntington disease. TBZ should be dosed individually based on ongoing assessment of possible adverse side effects. PMID- 16476935 TI - Autoantibody targeting of brain and intestinal transglutaminase in gluten ataxia. AB - OBJECTIVE: To investigate the presence of autoantibody deposition against type 2 tissue transglutaminase (TG2; a reliable marker of the whole spectrum of gluten sensitivity) in the jejunal tissue and brain of patients with gluten ataxia and in control subjects. METHODS: The authors evaluated jejunal biopsy samples from nine patients with gluten ataxia and seven patients with other causes of ataxia for the presence of TG2-related immunoglobulin deposits using double-color immunofluorescence. Autopsy brain tissue from one patient with gluten ataxia and one neurologically intact brain were also studied. RESULTS: IgA deposition on jejunal TG2 was found in the jejunal tissue of all patients with gluten ataxia and in none of the controls. The intestinal IgA deposition pattern was similar to that seen in patients with overt and latent celiac disease and in those with dermatitis herpetiformis. Widespread IgA deposition around vessels was found in the brain of the patient with gluten ataxia but not the control brain. The deposition was most pronounced in the cerebellum, pons, and medulla. CONCLUSIONS: Anti-tissue transglutaminase IgA antibodies are present in the gut and brain of patients with gluten ataxia with or without an enteropathy in a similar fashion to patients with celiac disease, latent celiac disease, and dermatitis herpetiformis but not in ataxia control subjects. This finding strengthens the contention that gluten ataxia is immune mediated and belongs to the same spectrum of gluten sensitivity as celiac disease and dermatitis herpetiformis. PMID- 16476936 TI - Involvement of vagal autonomic nuclei in multiple system atrophy and Lewy body disease. AB - BACKGROUND: Multiple system atrophy (MSA) and Lewy body disorders (LBDs) are associated with impaired control of gastrointestinal and cardiac functions. The dorsal vagal nucleus (DMV) innervates enteric neurons, whereas the ventrolateral nucleus ambiguus (NAmb) innervates the heart. The relationship between DMV and NAmb involvement and the gastrointestinal or cardiovagal manifestations in MSA and LBD is unclear. METHODS: The authors counted the cholinergic neurons in the DMV and NAmb in 15 cases of neuropathologically confirmed MSA, 14 of LBD (4 brainstem, 3 limbic, and 7 neocortical), and 12 control cases. All MSA and 8 of the 14 LBD cases had gastrointestinal symptoms; 8 of 12 MSA and 1 of 4 LBD cases had laboratory evidence of cardiovagal failure; 5 of the MSA and no LBD cases had laryngeal stridor. RESULTS: There was loss of cholinergic DMV neurons in all MSA and LBD cases. The degree of DMV cell loss was similar in LBD patient with or without gastrointestinal symptoms. In MSA but not in LBD cases, there was neuronal loss in the ventrolateral NAmb, with lower counts in patients with cardiovagal failure. CONCLUSIONS: There is comparable involvement of the dorsal vagal nucleus (DMV) in multiple system atrophy (MSA) and different stages of Lewy body disorders (LBDs). The relationship of DMV involvement and gastrointestinal symptoms is uncertain. Loss of neurons in the ventrolateral nucleus ambiguus may explain the more consistent cardiovagal failure in MSA than in LBD. PMID- 16476937 TI - Clinical correlates of cerebral water diffusion in Wilson disease. AB - OBJECTIVE: To investigate the course of diffusion changes in Wilson disease (WD) and to evaluate their clinical and radiologic correlates. METHODS: MRI with fluid attenuated inversion recovery (FLAIR) and diffusion weighted images (DWI) were performed in 13 symptomatic patients with WD who had typical neurologic manifestations of the disease (sWD patients) and in 5 presymptomatic patients (psWD patients). Follow-up clinical and MRI data were obtained in 12/13 patients with sWD. Ten subjects without neurologic disease and with normal cerebral MRI were used as controls. Apparent diffusion coefficient (ADC) was measured in areas where hyperintense lesions were detected on FLAIR images and in the normal appearing white matter. RESULTS: Hyperintense lesions were detected in all symptomatic patients on FLAIR MR images but only in 11 of 13 patients with sWD on DWI. These lesions were absent in patients with psWD. The mean ADC was found increased in the putamen, pallidum, internal capsule, mesencephalon, and within the white matter in the symptomatic group in comparison to controls. This was not observed in patients with psWD, who even had a decreased ADC in the putamen. A significant correlation was found between the increase in diffusion and the modified Rankin Scale in presence of symptoms. Moreover, the variation of the clinical scale was significantly correlated with the variation of diffusion in the putamen of symptomatic patients. CONCLUSION: A decrease in diffusion in the putamen can be detected before the occurrence of neurologic manifestations in WD. In contrast, a large increase in diffusion is detected after the occurrence of symptoms within the putamen, pallidum, internal capsule, and subcortical white matter parallel to the signal changes as seen on fluid-attenuated inversion recovery and diffusion weighted images. PMID- 16476938 TI - Hyperacusis in Williams syndrome: characteristics and associated neuroaudiologic abnormalities. AB - BACKGROUND: Hyperacusis and phonophobia are common, debilitating symptoms in Williams syndrome (WS), yet little is known about their underlying audiologic and neurologic processes. METHODS: The mothers of 49 subjects with WS were asked to complete the Hyperacusis Screening Questionnaire. Subjects with reported hyperacusis and sufficient developmental capacity underwent comprehensive audiological and brain auditory evoked response (BAER) testing. Findings were compared with those from pair-matched typically developing control subjects. RESULTS: Forty-one of the 49 children with WS (84%) had hyperacusis of moderate to severe degree, which began in infancy. Of these, 21 (mean age 15.8 +/- 5.5 years) were quantitatively tested. Subjects with WS reported discomfort at sound intensities on average 20 dB lower than control subjects. Pure-tone audiometry and distortion products otoacoustic emission test revealed a high-frequency cochlear hearing loss. An absence of ipsilateral acoustic reflex responses to maximum stimulation was significantly more common in the subjects with WS than controls. On BAER testing, the WS group had a significant prolongation in wave I latency. CONCLUSIONS: Hyperacusis in Williams syndrome (WS) is associated with a high-frequency hearing loss resembling the configuration of noise-induced hearing loss. The hyperacusis and hearing loss in WS may stem from a deficiency in the acoustic reflex resulting from auditory nerve dysfunction. Additional mechanisms that may mediate hyperacusis in WS and should be evaluated in future studies include recruitment, malformation of the facial canal, and haploinsufficiency of the elastin gene. PMID- 16476940 TI - The neurologic content of S. Weir Mitchell's fiction. AB - BACKGROUND: Silas Weir Mitchell (1829 to 1914), one of the most important neurologists in American Medicine, was known for his seminal work on the phantom limb syndrome, causalgia, and nerve injuries. He was also a prolific writer of novels and short stories. The neurologic content of this fiction has not been studied. OBJECTIVE: To assess the extent that references to neurologic topics were present in Mitchell's fiction, whether these neurologic references reflected Mitchell's scientific interests and contributions, and whether his fictional accounts of neurologic topics would precede those in his scientific writings. METHODS: The authors read Silas Weir Mitchell's novels and short stories. RESULTS: Seventeen (63.0%) of 27 fictional works contained neurologic references. Fifty-five (69.6%) of 79 references were brief (a single word or sentence). In two works, a neurologic theme was central to the plot. Some of the neurologic content was sophisticated (aphasia, brain laterality). Phantom limb syndrome, causalgia, and nerve injuries were not prominent in his fiction. Neurologic consequences of battle injuries were featured in 10 (37.0%) works. With the exception of "The Case of George Dedlow" (i.e., phantom limb syndrome), Mitchell's fictional accounts of neurologic topics followed his presentation of these topics in the scientific literature. CONCLUSIONS: The majority of Mitchell's fictional works contained references to neurologic topics but most contained brief references. The number of references to Mitchell's specific scientific interests (phantom limb syndrome, causalgia) was small, although more generally, references to the neurology of battle injuries occurred more frequently. PMID- 16476939 TI - Phenotypic and cellular expression of two novel connexin32 mutations causing CMT1X. AB - OBJECTIVE: To determine the phenotypic and cellular expression of two novel connexin32 (Cx32) mutations causing X-linked Charcot-Marie-Tooth disease (CMT1X). METHODS: The authors evaluated several members of two families with CMT1X clinically, electrophysiologically, pathologically, and by genetic testing. The Cx32 mutations were expressed in vitro and studied by immunocytochemistry. RESULTS: In both families, men were more severely affected than women with onset in the second decade of life. In the first family, the phenotype was that of demyelinating polyneuropathy with variable involvement of peripheral nerves. There was clinical evidence of CNS involvement in at least three of the patients, with extensor plantar responses and brisk reflexes. In the second family, the affected man presented with symmetric polyneuropathy and intermediate slowing of conduction velocities, whereas affected women had prominent asymmetric atrophy of the leg muscles. The authors identified two novel missense mutations resulting in L143P amino acid substitution in the first family and in V140E substitution in the second family, both located in the third transmembrane domain of Cx32. Expression of these Cx32 mutations in communication-incompetent HeLa cells and immunocytochemical analysis revealed that both mutants were retained intracellularly and were localized in the Golgi apparatus. In contrast to wild type protein, they did not form gap junctions. CONCLUSION: These novel connexin32 (Cx32) mutations cause a spectrum of clinical manifestations characteristic of Charcot-Marie-Tooth disease (CMT1X), including demyelinating or intermediate polyneuropathy, which is often asymmetric, and CNS involvement in one family. The position and cellular expression of Cx32 mutations alone cannot fully predict these phenotypic variations in CMT1X. PMID- 16476941 TI - Randomized placebo-controlled study of the nicotinic agonist SIB-1508Y in Parkinson disease. AB - This randomized, placebo-controlled, 5-week Phase II trial evaluated the safety and tolerability of SIB-1508Y, a selective alpha4beta2 nicotinic acetylcholine receptor agonist, in 77 individuals with early Parkinson disease. Lightheadedness was a common dosage-related adverse effect at higher dosages, leading to frequent dosage reduction, drug discontinuation, and eventual trial redesign. A maximally tolerated dosage of 10 mg daily was identified. No antiparkinsonian or cognitive enhancing effects were demonstrated in this trial. PMID- 16476942 TI - Primary progressive freezing gait: a syndrome with many causes. AB - Primary progressive freezing gait (PPFG) is characterized by early gait freezing and a stereotyped progression. Of nine patients followed up for 6 to 16 years, two were diagnosed pathologically: pallidonigroluysian degeneration (PNLD) and diffuse Lewy body disease. Four others evolved clinically into progressive supranuclear palsy and corticobasal degeneration. PPFG is not a distinct disorder but a syndrome with diverse causes. Long-term follow-up (> or =10 years) and postmortem are required for accurate diagnosis. PNLD may be the primary form of disease. PMID- 16476943 TI - Glucocerebrosidase gene mutations and Parkinson disease in the Norwegian population. AB - An association between mutations in the glucocerebrosidase (GBA) gene and Parkinson disease (PD) was recently reported in Ashkenazi Jews. The authors screened a series of 311 Norwegian patients with PD and 474 controls for 2 common functional mutations of the GBA protein, N370S and L444P. Seven patients (2.3%) and 8 controls (1.7%) carried a mutant GBA allele (p = 0.58). This study does not indicate increased susceptibility to PD in GBA mutations carriers in Norway. PMID- 16476945 TI - Unexpected pathogenic mechanism of a novel mutation in the coding sequence of SPG4 (spastin). AB - The authors report a nucleotide substitution (c.1216A>G) in SPG4 (spastin) causing hereditary spastic paraplegia. This apparent missense mutation in the ATPase domain confers aberrant, in-frame splicing and results in destabilization of mutated transcript. Mutated protein is deficient in microtubule-severing activity but, unlike neighboring mutations, shows regular subcellular localization. The authors' data point to haploinsufficiency rather than a dominant negative effect as the disease-causing mechanism for this mutation. PMID- 16476944 TI - Oscillatory pallidal local field potential activity correlates with involuntary EMG in dystonia. AB - The pathophysiology of dystonia is unclear. The authors recorded local field potentials (LFPs) from deep brain stimulation electrodes implanted in the pallidum of 13 dystonic patients. LFP power correlated with the level of dystonic EMG in the sternocleidomastoid, with maximal positive correlations at the lower contacts of pallidal electrodes. The data suggest that the neuronal synchronization indexed by LFP oscillations in the globus pallidus may be mechanistically linked to dystonic EMG activity. PMID- 16476946 TI - Pontine and cerebellar atrophy correlate with clinical disability in SCA2. AB - Spinocerebellar ataxia type 2 (SCA2) has protean manifestations, and a clinical marker of progression is needed. Although MRI is a promising tool, it is unclear whether the degree of atrophy shown on MRI is correlated with clinical dysfunction. Here the authors used high-resolution volumetric MRI analysis to show that cerebellar and pontine volumes specifically and closely correlate with functional staging scores. PMID- 16476948 TI - Silent microbleeds are associated with volume of primary intracerebral hemorrhage. AB - The authors performed a correlative radiologic study on the micro-bleeds and volume of intracerebral hemorrhage in the supratentorial ICH patients. In the patients with lobar or putaminal hemorrhage, the hemorrhage volumes increased more than twofold or threefold in the patients with micro-bleeds. Moreover, the presence of microbleeds was an independent risk factor for large-sized hemorrhage. These data show that microbleeds may be associated with a larger ICH volume. PMID- 16476947 TI - Is protracted low-dose temozolomide feasible in glioma patients? AB - The authors investigated the safety of 75 mg/m2 temozolomide for 21 days every 28 days in glioma patients. This schedule could lead to DNA repair enzyme O6 alkylguanine-DNA alkyltransferase depletion, contributing to overcoming drug resistance. Although Phase III studies are forthcoming, no data are available on the long-term toxicity of temozolomide, which, in this series, incurred prolonged, cumulative lymphopenia, which leads to a high incidence of infections. PMID- 16476949 TI - Vascular and Alzheimer-type pathology in an autopsy study of African-Americans. AB - The authors studied 13 autopsy brains from a larger cohort of 270 African Americans with a clinical diagnosis of Alzheimer disease (AD), vascular dementia (VaD), or stroke without dementia. Two subjects exhibited changes of pure VaD, 5 had pure AD, and 6 showed a mixture of AD pathology and strokes. Overall, there was good agreement between the pathologic diagnoses and the clinical diagnoses. PMID- 16476950 TI - Creutzfeldt-Jakob disease among American Indians and Alaska Natives in the United States. AB - The occurrence of Creutzfeldt-Jakob disease (CJD) among American Indians and Alaska Natives in the United States was evaluated using national multiple cause of-death data and medical information obtained from state health departments. Twelve CJD deaths were identified for 1981 through 2002, and the average annual age-adjusted death rate was 0.47 per million population. This rate was significantly lower than that for whites and similar to the rate for African Americans. PMID- 16476951 TI - Fascicular hypoglossus nerve lesion. PMID- 16476952 TI - Methionine metabolism and phenotypic variability in X-linked adrenoleukodystrophy. AB - A combined genotype of polymorphisms of methionine metabolism has been associated with CNS demyelination in methotrexate-treated patients. Within a sample of 86 patients with X-linked adrenoleukodystrophy, this genotype was overrepresented in a subgroup of 15 patients with adrenomyeloneuropathy (AMN) with CNS demyelination (adrenoleukomyeloneuropathy) in comparison to 49 AMN patients without CNS demyelination ("pure" AMN; p = 0.002), suggesting that methionine metabolism might contribute to the phenotypic variability in adrenoleukodystrophy. PMID- 16476953 TI - Multiplex analysis of expression of three IFNbeta-induced genes in antibody positive MS patients. AB - Some interferon beta (IFNbeta)-treated patients with multiple sclerosis develop antibody-mediated decreased bioactivity with resultant loss of therapeutic effect. The authors developed real-time multiplex reverse transcriptase PCR to measure expression of three IFNbeta-inducible genes to directly assess IFNbeta bioactivity in patients with neutralizing antibodies (NAbs). The three genes responded in tandem. Correlation of NAb level with bioactivity at low/moderate NAb levels was poor, indicating that for such patients, direct measurement of IFNbeta bioactivity is most reliable. PMID- 16476954 TI - Pure myopathy associated with a novel mitochondrial tRNA gene mutation. AB - The authors describe a 47-year-old man who presented with proximal muscle weakness, myalgia, elevated creatine kinase, and features of a pure myopathic syndrome in whom they have identified a novel mutation in the mitochondrial tRNA(Ala) gene. This 5591G>A transition is heteroplasmic, segregates with cytochrome c oxidase deficiency in single muscle fibers, and fulfills recognized criteria for pathogenicity. This case exemplifies the wide-ranging clinical spectrum of mitochondrial disease presentations. PMID- 16476955 TI - Paraneoplastic encephalomyelitis associated with pancreatic tumor and anti-GAD antibodies. PMID- 16476956 TI - Lower motor neuron weakness after diving-related decompression. PMID- 16476957 TI - Cortical liquefaction in severe human herpesvirus 6 encephalopathy. PMID- 16476958 TI - Glomeruloid hemangiomas: a marker for POEMS. PMID- 16476960 TI - Partial retraction of correspondence about "the pentapeptide QYNAD does not block voltage-gated sodium channels". PMID- 16476959 TI - Acute trismus associated with Foix-Marie-Chavany syndrome. PMID- 16476961 TI - Brain Mets: a poem in three voices. PMID- 16476962 TI - Searching for a relationship between manganese and welding and Parkinson's disease. PMID- 16476963 TI - Language cortex activation in normal children. PMID- 16476964 TI - The smoking-thrombolysis paradox and acute ischemic stroke. PMID- 16476965 TI - Treatment of carpal tunnel syndrome: is there a role for local corticosteroid injection? PMID- 16476966 TI - Association of the metabolic syndrome with intracranial atherosclerotic stroke. PMID- 16476967 TI - The first organ transplant from a brain-dead donor. PMID- 16476968 TI - Prospective reliability of the STRokE DOC Wireless/Site Independent Telemedicine System. PMID- 16476969 TI - An RNA aptamer that distinguishes between closely related human influenza viruses and inhibits haemagglutinin-mediated membrane fusion. AB - Aptamers selected against various kinds of targets have shown remarkable specificity and affinity, similar to those displayed by antibodies to their antigens. To employ aptamers as genotyping reagents for the identification of pathogens and their strains, in vitro selections were carried out to find aptamers that specifically bind and distinguish the closely related human influenza A virus subtype H3N2. The selected aptamer, P30-10-16, binds specifically to the haemagglutinin (HA) region of the target strain A/Panama/2007/1999(H3N2) and failed to recognize other human influenza viruses, including another strain with the same subtype, H3N2. The aptamer displayed over 15-fold-higher affinity to the HA compared with the monoclonal antibody, and efficiently inhibited HA-mediated membrane fusion. These studies delineate the application of aptamers in the genotyping of viruses. PMID- 16476970 TI - Abnormal immune response of CCR5-deficient mice to ocular infection with herpes simplex virus type 1. AB - Ocular herpes simplex virus type 1 (HSV-1) infection elicits a strong inflammatory response that is associated with production of the beta chemokines CCL3 and CCL5, which share a common receptor, CCR5. To gain insight into the role of these molecules in ocular immune responses, the corneas of wild-type (WT) and CCR5-deficient (CCR5-/-) mice were infected with HSV-1 and inflammatory parameters were measured. In the absence of CCR5, the early infiltration of neutrophils into the cornea was diminished. Associated with this aberrant leukocyte recruitment, neutrophils in CCR5-/- mice were restricted to the stroma, whereas in WT mice, these cells trafficked to the stroma and epithelial layers of the infected cornea. Virus titres and cytokine/chemokine levels in the infected tissue of these mice were similar for the first 5 days after infection. However, by day 7 post-infection, the CCR5-/- mice showed a significant elevation in the chemokines CCL2, CCL5, CXCL9 and CXCL10 in the trigeminal ganglion and brainstem, as well as a significant increase in virus burden. The increase in chemokine expression was associated with an increase in the infiltration of CD4 and/or CD8 T cells into the trigeminal ganglion and brainstem of CCR5-/- mice. Surprisingly, even though infected CCR5-/- mice were less efficient at controlling the progression of virus replication, there was no difference in mortality. These results suggest that, although CCR5 plays a role in regulating leukocyte trafficking and control of virus burden, compensatory mechanisms are involved in preventing mortality following HSV-1 infection. PMID- 16476971 TI - Human herpesvirus 7 U47 gene products are glycoproteins expressed in virions and associate with glycoprotein H. AB - The function of the human herpesvirus 7 (HHV-7) U47 gene, which is a positional homologue of the genes encoding glycoprotein O (gO) in human cytomegalovirus (HCMV) and human herpesvirus 6 (HHV-6), was analysed. A monoclonal antibody (mAb) against the U47 gene product reacted in immunoblots with proteins migrating at 49 and 51 kDa in lysates of HHV-7-infected cells and with 49 and 51 kDa proteins in partially purified virions. Digestion of the 49 and 51 kDa proteins with endoglycosidase H and peptide N-glycosidase F indicated that the U47-encoded proteins were modified with N-linked oligosaccharides. Therefore, the U47 gene and its product were named gO, as in HCMV and HHV-6. In addition, the anti-gO mAb co-immunoprecipitated glycoprotein H (gH) in HHV-7-infected cells, indicating an association between HHV-7 gO and gH. The results suggest that the HHV-7 gO-gH complex might have a similar function to that in HCMV or HHV-6, such as cell-cell fusion in virus infection. PMID- 16476972 TI - Cloning of the genome of Alcelaphine herpesvirus 1 as an infectious and pathogenic bacterial artificial chromosome. AB - Alcelaphine herpesvirus 1 (AlHV-1), carried asymptomatically by wildebeest, causes malignant catarrhal fever (MCF) following cross-species transmission to a variety of susceptible species of the order Artiodactyla. The study of MCF pathogenesis has been impeded by an inability to produce recombinant virus, mainly due to the fact that AlHV-1 becomes attenuated during passage in culture. In this study, these difficulties were overcome by cloning the entire AlHV-1 genome as a stable, infectious and pathogenic bacterial artificial chromosome (BAC). A modified loxP-flanked BAC cassette was inserted in one of the two large non-coding regions of the AlHV-1 genome. This insertion allowed the production of an AlHV-1 BAC clone stably maintained in bacteria and able to regenerate virions when transfected into permissive cells. The loxP-flanked BAC cassette was excised from the genome of reconstituted virions by growing them in permissive cells stably expressing Cre recombinase. Importantly, BAC-derived AlHV-1 virions replicated comparably to the virulent (low-passage) AlHV-1 parental strain and induced MCF in rabbits that was indistinguishable from that of the virulent parental strain. The availability of the AlHV-1 BAC is an important advance for the study of MCF that will allow the identification of viral genes involved in MCF pathogenesis, as well as the production of attenuated recombinant candidate vaccines. PMID- 16476973 TI - Identifying cellular genes crucial for the reactivation of Kaposi's sarcoma associated herpesvirus latency. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) is the latest addition to the long list of human herpesviruses. Reactivation of latent herpesvirus infections is still a mystery. It was demonstrated recently that the phorbol ester TPA was efficient in inducing a reactivation of KSHV infection in the S phase of the cell cycle. In the present study, flow cytometry-sorted, TPA-induced, KSHV-infected haematopoietic cells (BCBL-1) were used to analyse the expression profiles of cancer-related cellular genes in the S phase of the cell cycle compared with the G0/1 phase by using microarrays. Overall, the S phase of the cell cycle seems to provide KSHV with an apt environment for a productive lytic cycle of infection. The apt conditions include cellular signalling that promotes survivability, DNA replication and lipid metabolism, while blocking cell-cycle progression to M phase. Some of the important genes that were overexpressed during the S phase of the cell cycle compared with the G0/1 phase of TPA-induced BCBL-1 cells are v-myb myeloblastosis (MYBL2), protein kinase-membrane associated tyrosine/threonine 1 (PKMYT1), ribonucleotide reductase M1 polypeptide (RRM1) and peroxisome proliferator-activated receptors delta (PPARD). Inhibition of PKMYT1 expression by the use of specific short interfering RNAs significantly lowered the TPA induced KSHV lytic cycle of infection. The significance of these and other genes in the reactivation of KSHV is discussed in the following report. Taken together, a flow cytometry-microarray-based method to study the cellular conditions critical for the reactivation of KSHV infection is reported here for the first time. PMID- 16476974 TI - Dual mutations in the Autographa californica nucleopolyhedrovirus FP-25 and p35 genes result in plasma-membrane blebbing in Trichoplusia ni cells. AB - Spodoptera frugiperda cells infected with Autographa californica nucleopolyhedrovirus (AcMNPV) lacking a functional anti-apoptotic p35 protein undergo apoptosis. However, such mutants replicate normally in Trichoplusia ni (TN-368) cells. An AcMNPV plaque isolate (AcdefrT) was identified during propagation of a virus deficient in p35 in TN-368 cells. This virus exhibited enhanced budded-particle formation in TN-368 cells, but was partially defective for polyhedra production in the same cells. Virus replication in AcdefrT-infected TN-368 cells was accompanied by extensive plasma-membrane blebbing and caspase activation late in infection, both features of apoptosis. Rescue of the p35 locus of AcdefrT continued to result in a reduction in polyhedra and increase in budded virus production in TN-368 cells, but no plasma-membrane blebbing was observed. The mutation was mapped to the FP-25 gene locus. This gene mutation combined with the non-functional p35 was found to be responsible for the cell-blebbing effect observed in AcdefrT-infected TN-368 cells. PMID- 16476975 TI - Genome sequence of an enhancin gene-rich nucleopolyhedrovirus (NPV) from Agrotis segetum: collinearity with Spodoptera exigua multiple NPV. AB - The genome sequence of a Polish isolate of Agrotis segetum nucleopolyhedrovirus (AgseNPV-A) was determined and analysed. The circular genome is composed of 147,544 bp and has a G+C content of 45.7 mol%. It contains 153 putative, non overlapping open reading frames (ORFs) encoding predicted proteins of more than 50 aa, together making up 89.8 % of the genome. The remaining 10.2 % of the DNA constitutes non-coding regions and homologous-repeat regions. One hundred and forty-three AgseNPV-A ORFs are homologues of previously reported baculovirus gene sequences. There are ten unique ORFs and they account for 3 % of the genome in total. All 62 lepidopteran baculovirus genes, including the 29 core baculovirus genes, were found in the AgseNPV-A genome. The gene content and gene order of AgseNPV-A are most similar to those of Spodoptera exigua (Se) multiple NPV and their shared homologous genes are 100 % collinear. Three putative enhancin genes were identified in the AgseNPV-A genome. In phylogenetic analysis, the AgseNPV-A enhancins form a cluster separated from enhancins of the Mamestra species NPVs. PMID- 16476976 TI - Relaxed template specificity in fowl adenovirus 1 DNA replication initiation. AB - The fowl adenovirus 1 (FAdV-1) isolates PHELPS and OTE are highly similar, but have striking differences in the repeat region of the inverted terminal repeat (ITR). Whilst the repeat region in OTE conforms to the conventional human adenovirus repeat region (5'-CATCATC), that of PHELPS contains guanidine residues at positions 1, 4 and 7 (5'-GATGATG). This implies that the FAdV-1 isolates PHELPS and OTE have either distinct template specificity at replication initiation or, alternatively, a relaxed specificity for replication initiation. In this study, the distinct sequence variation at the origin of DNA replication in the ITRs of the FAdV-1 PHELPS and OTE isolates was confirmed. Sequence analyses of the pTP and Pol genes of both PHELPS and OTE did not reveal differences that could explain the distinct template specificity. Replication assays demonstrated that linear DNA fragments flanked by either 5'-CATCATC or 5' GATGATG termini replicated in cells upon infection with FAdV-1 OTE and FAdV-1 PHELPS. This was evident from the appearance of DpnI-resistant fragments in a minireplicon assay. From these data, it is concluded that FAdV-1 has relaxed, rather than changed, its template specificity at replication initiation. PMID- 16476977 TI - Role of myristoylation and N-terminal basic residues in membrane association of the human immunodeficiency virus type 1 Nef protein. AB - Human immunodeficiency virus type 1 Nef protein is N-terminally myristoylated, a modification reported to be required for the association of Nef with cytoplasmic membranes. As myristate alone is not sufficient to anchor a protein stably into a membrane, it has been suggested that N-terminal basic residues contribute to Nef membrane association via electrostatic interactions with acidic phospholipids. Here, data are presented pertaining to the role of the myristate and basic residues in Nef membrane association, subcellular localization and function. Firstly, by using a biochemical assay for membrane association it was shown that, whereas myristoylation of Nef was not essential, mutation of a cluster of four arginines between residues 17 and 22 reduced membrane association dramatically. Mutation of two lysines at residues 4 and 7 had negligible effect alone, but when combined with the arginine substitutions, abrogated membrane association completely. By using indirect immunofluorescence, it was demonstrated that mutation of either of the two basic clusters altered the subcellular distribution of Nef dramatically. Thirdly, the requirement of the arginine and lysine clusters for Nef-mediated CD4 down modulation was shown to correlate precisely with membrane association. These data suggest that membrane localization and subcellular targeting of Nef are controlled by a complex interplay of signals at the N terminus of the protein. PMID- 16476978 TI - Genetic diversity of small-ruminant lentiviruses: characterization of Norwegian isolates of Caprine arthritis encephalitis virus. AB - Small-ruminant lentiviruses (SRLVs), including Caprine arthritis encephalitis virus (CAEV) in goats and maedi-visna virus (MVV) in sheep, are lentiviruses that, despite overall similarities, show considerable genetic variation in regions of the SRLV genome. To gain further knowledge about the genetic diversity and phylogenetic relationships among field isolates of SRLVs occurring in geographically distinct areas, the full-length genomic sequence of a CAEV isolate (CAEV-1GA) and partial env sequences obtained from Norwegian CAEV-infected goats were determined. The genome of CAEV-1GA consisted of 8,919 bp. Alignment studies indicated significant diversity from published SRLV sequences. Deletions and hypervariability in the 5' part of the env gene have implications for the size of the proposed CAEV-1GA Rev protein and the encoded surface glycoprotein (SU). The variable regions in the C-terminal part of SU obtained from Norwegian CAEV isolates demonstrate higher sequence divergence than has been described previously for SRLVs. Phylogenetic analysis based on SU sequences gives further support for a unique group designation. The results described here reveal a distant genetic relationship between Norwegian CAEV and other SRLVs and demonstrate that there is more geographical heterogeneity among SRLVs than reported previously. PMID- 16476979 TI - Comparative studies on mucosal and intravenous transmission of simian immunodeficiency virus (SIVsm): evolution of coreceptor use varies with pathogenic outcome. AB - Coreceptor usage of isolates from 30 cynomolgus macaques infected intrarectally (n=22) or intravenously (n=8) with simian immunodeficiency virus of sooty mangabey origin (SIVsm) was evaluated in U87.CD4 and GHOST(3) cell lines. Based on progression rate, the animals were divided into progressors (18 animals), slow progressors (five animals) and long-term non-progressors (seven animals). There was no difference in how many or which coreceptors were used according to route of infection. All isolates but one used CCR5 for cell entry, and CCR5 was also the major coreceptor in 70 out of 105 isolates tested. In general, early isolates were multitropic, using CCR5, CXCR6 and/or gpr15. Interestingly, CXCR4-using viruses could be isolated on human peripheral blood mononuclear cells (PBMCs), but not on cynomolgus macaque PBMCs, suggesting that human PBMCs select for variants with CXCR4 use. Even though CXCR4-using SIV isolates have been reported rarely, we could recover CXCR4-using viruses from 13 monkeys. CXCR4 use either appeared early during the acute phase of infection and disappeared later or only appeared late in infection during immunodeficiency. Surprisingly, one late isolate from a progressor monkey did not use CCR5 at all and used the CXCR4 receptor with high efficiency. The ability to use many different receptors decreased over time in long-term non-progressor monkeys, whilst the majority of progressor monkeys showed broadening of coreceptor use, stable coreceptor use or fluctuation between the different coreceptor-usage patterns. The results indicate that, in the infected host, evolution of SIV coreceptor usage occurs, involving changes in the mode of coreceptor use. PMID- 16476980 TI - Comparative studies on mucosal and intravenous transmission of simian immunodeficiency virus (SIVsm): the kinetics of evolution to neutralization resistance are related to progression rate of disease. AB - The kinetics of appearance of autologous neutralizing antibodies were studied in cynomolgus macaques infected with simian immunodeficiency virus (SIVsm) by the intravenous (IV) route (six monkeys) or the intrarectal (IR) route (ten monkeys). The SIVsm inoculum virus and reisolates obtained at 2 weeks, 3 or 4 months and later than 1 year were tested in a GHOST(3) cell line-based plaque-reduction assay with autologous sera collected at the same sampling times. All monkeys developed a neutralizing-antibody response to the inoculum virus, those infected by the IV route earlier than monkeys infected by the IR route. Animals were divided into progressor (P), slow-progressor (SP) and long-term non-progressor (LTNP) monkeys, based on progression rate. In P monkeys, neutralization escape could be demonstrated by 3 months post-infection. Neutralization-resistant variants also emerged in SP and LTNP monkeys, but were much delayed compared with P monkeys. Evolution of neutralization resistance was also demonstrated by a positive-control serum in the heterologous reaction. Pooled sera from four LTNP monkeys showed a broad neutralizing capacity, including neutralization of escape variants. These results from a large group of infected monkeys showed that SIV evolves to neutralization resistance in the infected host and that the kinetics of this evolution are related to the route of transmission and the progression rate of SIV disease. The results suggest an important role for neutralizing antibodies in controlling viraemia. Although this control is transient in the infected host, neutralization resistance is relative and variant viruses may be neutralized by a broadly cross-neutralizing serum pool. PMID- 16476981 TI - CCR5 use by human immunodeficiency virus type 1 is associated closely with the gp120 V3 loop N-linked glycosylation site. AB - Human immunodeficiency virus type 1 (HIV-1) enters cells through the chemokine receptors CCR5 (R5 virus) and/or CXCR4 (X4 virus). Loss of N-linked glycans and increased net charge of the third variable loop (V3) of the gp120 envelope glycoprotein have been observed to be important steps towards CXCR4 use. All reported sequences using CCR5 or CXCR4 exclusively, or using both, were gathered from the Los Alamos HIV Database and analysed with regard to the V3 N-linked glycosylation motifs (sequons) and charge. The V3 loop glycan had a sensitivity of 0.98 and a 0.92 positive predictive value in the context of CCR5 use. The difference from X4 was remarkable (P<10(-12)). Especially, the sequon motif NNT within the V3 loop was conserved in 99.2 % of the major clades. The results suggest a close association between the V3 loop glycan and CCR5 use and may provide new insight into HIV-1 tropism and help to improve phenotype-prediction models. PMID- 16476982 TI - The glycosylation site in the envelope protein of West Nile virus (Sarafend) plays an important role in replication and maturation processes. AB - The complete genome of West Nile (Sarafend) virus [WN(S)V] was sequenced. Phylogenetic trees utilizing the complete genomic sequence, capsid gene, envelope gene and NS5 gene/3' untranslated region of WN(S)V classified WN(S)V as a lineage II virus. A full-length infectious clone of WN(S)V with a point mutation in the glycosylation site of the envelope protein (pWNS-S154A) was constructed. Both growth kinetics and the mode of maturation were affected by this mutation. The titre of the pWNS-S154A virus was lower than the wild-type virus. This defect was corrected by the expression of wild-type envelope protein in trans. The pWNS S154A virus matured intracellularly instead of at the plasma membrane as shown for the parental WN(S)V. PMID- 16476983 TI - Efficient cleavage by signal peptide peptidase requires residues within the signal peptide between the core and E1 proteins of hepatitis C virus strain J1. AB - Maturation of hepatitis C virus (HCV) core protein requires cleavage by signal peptidase (SP) and signal peptide peptidase (SPP) at a signal peptide between core and the E1 glycoprotein. For HCV strain Glasgow, amino acids Ala(180), Ser(183) and Cys(184) within the signal peptide have previously been shown to be essential for efficient SPP cleavage. By contrast, these residues apparently did not contribute to core maturation in HCV strain J1. In the present study, the source of this discrepancy has been analysed and it is concluded that interpretation of the strain J1 data was incorrect, due to the inability to separate wild-type and mutant forms of core on gels by using standard buffer systems. PMID- 16476985 TI - Tagging of NS5A expressed from a functional hepatitis C virus replicon. AB - Knowledge of how hepatitis C virus (HCV) proteins associate with components of the host cell to form a functional replication complex is still limited. To address this issue, HCV replicon constructs were generated where either green fluorescent protein (GFP) or the Propionibacterium shermanii transcarboxylase domain (PSTCD) was introduced into the NS5A coding region. Insertion of both GFP and PSTCD was tolerated well, allowing formation of stable replicon-containing cell lines that contained viral protein and transcript levels that were comparable to those of an unmodified parental replicon. Cell lines generated from the GFP-tagged NS5A replicon allowed live-cell visualization of the location of NS5A. Cell lines generated from the PSTCD-tagged replicons allowed rapid and efficient precipitation of the PSTCD-tagged NS5A, as well as other HCV non structural proteins, using streptavidin-coated magnetic beads. Both replicons represent useful tools that offer different but complementary ways of examining replication-complex formation in cells. PMID- 16476984 TI - Hepatitis C virus complete genome sequences identified from China representing subtypes 6k and 6n and a novel, as yet unassigned subtype within genotype 6. AB - Here, the complete genome sequences for three hepatitis C virus (HCV) variants identified from China and belonging to genotype 6 are reported: km41, km42 and gz52557. Their entire genome lengths were 9430, 9441 and 9448 nt, respectively; the 5' untranslated regions (UTRs) contained 341, 342 and 339 nt, followed by single open reading frames of 9045, 9045 and 9057 nt, respectively; the 3' UTRs, up to the poly(U) tracts, were 41, 51 and 52 nt, respectively. Phylogenetic analyses showed that km41 is classified into subtype 6k and km42 into subtype 6n. Although gz52557 clustered distantly with subtype 6g, it appeared to belong to a distinct subtype. Analysis with 53 and 105 partial core and NS5B region sequences, respectively, representing 17 subtypes from 6a to 6q and three unassigned isolates of genotype 6 in co-analyses demonstrated that gz52557 was equidistant from all of these isolates, indicating that it belongs to a novel subtype. However, based on a recent consensus that three or more examples are required for a new HCV subtype designation, it is suggested that gz52557 remains unassigned to any subtype. PMID- 16476986 TI - Comparative evaluation of two severe acute respiratory syndrome (SARS) vaccine candidates in mice challenged with SARS coronavirus. AB - Two different severe acute respiratory syndrome (SARS) vaccine strategies were evaluated for their ability to protect against live SARS coronavirus (CoV) challenge in a murine model of infection. A whole killed (inactivated by beta propiolactone) SARS-CoV vaccine and a combination of two adenovirus-based vectors, one expressing the nucleocapsid (N) and the other expressing the spike (S) protein (collectively designated Ad S/N), were evaluated for the induction of serum neutralizing antibodies and cellular immune responses and their ability to protect against pulmonary SARS-CoV replication. The whole killed virus (WKV) vaccine given subcutaneously to 129S6/SvEv mice was more effective than the Ad S/N vaccine administered either intranasally or intramuscularly in inhibiting SARS-CoV replication in the murine respiratory tract. This protective ability of the WKV vaccine correlated with the induction of high serum neutralizing-antibody titres, but not with cellular immune responses as measured by gamma interferon secretion by mouse splenocytes. Titres of serum neutralizing antibodies induced by the Ad S/N vaccine administered intranasally or intramuscularly were significantly lower than those induced by the WKV vaccine. However, Ad S/N administered intranasally, but not intramuscularly, significantly limited SARS CoV replication in the lungs. Among the vaccine groups, SARS-CoV-specific IgA was found only in the sera of mice immunized intranasally with Ad S/N, suggesting that mucosal immunity may play a role in protection for the intranasal Ad S/N delivery system. Finally, the sera of vaccinated mice contained antibodies to S, further suggesting a role for this protein in conferring protective immunity against SARS-CoV infection. PMID- 16476987 TI - Identification of protease and ADP-ribose 1''-monophosphatase activities associated with transmissible gastroenteritis virus non-structural protein 3. AB - The replicase polyproteins, pp1a and pp1ab, of porcine Transmissible gastroenteritis virus (TGEV) have been predicted to be cleaved by viral proteases into 16 non-structural proteins (nsp). Here, enzymic activities residing in the amino-proximal region of nsp3, the largest TGEV replicase processing product, were characterized. It was shown, by in vitro translation experiments and protein sequencing, that the papain-like protease 1, PL1(pro), but not a mutant derivative containing a substitution of the presumed active-site nucleophile, Cys(1093), cleaves the nsp2|nsp3 site at (879)Gly|Gly(880). By using an antiserum raised against the pp1a/pp1ab residues 526-713, the upstream processing product, nsp2, was identified as an 85 kDa protein in TGEV-infected cells. Furthermore, PL1(pro) was confirmed to be flanked at its C terminus by a domain (called X) that mediates ADP-ribose 1''-phosphatase activity. Expression and characterization of a range of bacterially expressed forms of this enzyme suggest that the active X domain comprises pp1a/pp1ab residues Asp(1320)-Ser(1486). PMID- 16476988 TI - Role of a conserved tripeptide in the endodomain of Sindbis virus glycoprotein E2 in virus assembly and function. AB - Envelopment of Sindbis virus (SV) at the plasma membrane begins with the interaction of the E2 glycoprotein endodomain with a hydrophobic cleft in the surface of the pre-assembled nucleocapsid. The driving force for this budding event is thought to reside in this virus type-specific association at the surface of the cell. The specific amino acids involved in this interaction have not been identified; however, it has been proposed that a conserved motif (TPY) at aa 398 400 in the E2 tail plays a critical role in this interaction. This interaction has been examined with virus containing mutations at two positions in this conserved domain, T398A and Y400N. The viruses produced have very low infectivity (as determined by particle : p.f.u. ratios); however, there appears to be no defect in assembly, as the virus has wild-type density and electron microscopy shows assembled particles with no obvious aberrant structural changes. The loss of infectivity in the double mutant is accompanied by the loss of the ability to fuse cells after brief exposure to acid pH. These data support the idea that these residues are vital for production of infectious/functional virus; however, they are dispensable for assembly. These results, combined with other published observations, expand our understanding of the interaction of the E2 endodomain with the capsid protein. PMID- 16476989 TI - Transcribing paramyxovirus RNA polymerase engages the template at its 3' extremity. AB - For the non-segmented, negative-stranded RNA viruses, the mechanism controlling transcription or replication is still a matter of debate. To gain information about this mechanism and about the nature of the RNA polymerase involved, the length of an intervening sequence separating the 3' end of Sendai virus minigenomes and a downstream transcription-initiation signal was increased progressively. It was found that transcription, as measured by green fluorescent protein (GFP) expression, decreased progressively in proportion to the increase in length of the intervening sequence. GFP expression correlated well with the levels of GFP mRNA in the cells, as measured by quantitative primer extension and by RNase protection. Thus, mRNA transcription was inversely proportional to the length of the inserted sequence. These data are evidence that the RNA polymerase initiating transcription at the downstream transcription signal somehow sees the distance separating this signal and the template 3' extremity. Implication of this observation for the nature of the Sendai virus RNA polymerase and for the mechanism by which it synthesizes mRNAs or replication products is presented. PMID- 16476990 TI - Role of non-raft cholesterol in lymphocytic choriomeningitis virus infection via alpha-dystroglycan. AB - Dystroglycan (DG) is an extracellular matrix receptor necessary for the development of metazoans from flies to humans and is also an entry route for various pathogens. Lymphocytic choriomeningitis virus (LCMV), a member of the family Arenaviridae, infects by binding to alpha-DG. Here, the role of cholesterol lipid rafts in infection by LCMV via alpha-DG was investigated. The cholesterol-sequestering drugs methyl-beta-cyclodextrin (MbetaCD), filipin and nystatin inhibited the infectivity of LCMV selectively, but did not affect infection by vesicular stomatitis virus. Cholesterol loading after depletion with MbetaCD restored infectivity to control levels. DG was not found in lipid rafts identified with the raft marker ganglioside GM1. Treatment with MbetaCD, however, enhanced the solubility of DG. This may reflect the association of DG with cholesterol outside lipid rafts and suggests that association of DG with non-raft cholesterol is critical for infection by LCMV through alpha-DG. PMID- 16476991 TI - A new promoter-binding site in the PB1 subunit of the influenza A virus polymerase. AB - The influenza A virus RNA-dependent RNA polymerase consists of three subunits PB1, PB2 and PA. The 5' and 3' terminal sequences of the viral RNA (vRNA) form the viral promoter and are bound by the PB1 subunit. The putative promoter binding sites of the PB1 subunit have been mapped in previous studies but with contradictory results. The aim of the current study was to investigate the function of two evolutionary conserved regions in PB1 - from aa 233 to 249 and 269 to 281, which lie immediately N- and C-terminal, respectively, of a previously proposed binding site for the 3' end of the vRNA promoter. The previously proposed binding site extended from aa 249 to 256 and centred on two phenylalanine residues (F251 and F254). However, the fact that F251 is required for polymerase activity was not confirmed here. Instead, it was proposed that the 233-249 region contains a new 5' vRNA promoter-binding site, and arginine residues crucial for this activity were characterized. However, residues 269-281 were unlikely to be directly involved in promoter binding. These results are discussed in relation to the previous studies and a new model for vRNA promoter binding to the influenza RNA polymerase is presented. PMID- 16476992 TI - Structural and functional integrity of the coxsackievirus B3 oriR: spacing between coaxial RNA helices. AB - The enterovirus oriR is composed of two helices, X and Y, anchored by a kissing (K) interaction. For proper oriR function, certain areas of these helices should be specifically oriented towards each other. It was hypothesized that the single stranded nucleotides bridging the coaxial helices (Y-X and K-Y linkers) are important to determine this orientation. Spatial changes were introduced by altering the linker length between the helices of the coxsackievirus B3 oriR. Changing the linker lengths resulted in defective RNA replication, probably because of an altered oriR geometry. The identity of the linker residues also played a role, possibly because of sequence-specific ligand recognition. Although each point mutation altering the primary sequence of the Y-X spacer resulted in defective growth at 36 degrees C, the mutations had a wild-type phenotype at 39 degrees C, indicating a cold-sensitive phenotype. The results show that the intrinsic connection between oriR structure and function is fine-tuned by the spacing between the coaxial RNA helices. PMID- 16476993 TI - Putative neutralization epitopes and broad cross-genotype neutralization of Hepatitis E virus confirmed by a quantitative cell-culture assay. AB - Monolayers of Hep G2/C3A cells were inoculated with genotype 1 Hepatitis E virus (HEV) mixed with either anti-HEV or an appropriate control. After 5 or 6 days, cell monolayers were stained with anti-HEV and infected cells were identified by immunofluorescence microscopy and counted. Anti-HEV from vaccinated or infected rhesus monkeys neutralized the virus, as did mAbs that recognized epitopes on the C terminus of a recombinant vaccine protein. Antibodies were broadly cross reactive, since convalescent serum from animals infected with any one of the four mammalian genotypes all neutralized the genotype 1 virus. PMID- 16476994 TI - Cytological analysis of Saccharomyces cerevisiae cells supporting cymbidium ringspot virus defective interfering RNA replication. AB - The replicase proteins p33 and p92 of Cymbidium ringspot virus (CymRSV) were found to support the replication of defective interfering (DI) RNA in Saccharomyces cerevisiae cells. Two yeast strains were used, differing in the biogenesis of peroxisomes, the organelles supplying the membranous vesicular environment in which CymRSV RNA replication takes place in infected plant cells. Double-labelled immunofluorescence showed that both p33 and p92 replicase proteins localized to peroxisomes, independently of one another and of the presence of the replication template. It is suggested that these proteins are sorted initially from the cytosol to the endoplasmic reticulum and then to peroxisomes. However, only the expression of p33, but not p92, increased the number of peroxisomes and induced membrane proliferation. DI RNA replication occurred in yeast cells, as demonstrated by the presence of monomers and dimers of positive and negative polarities. Labelling with BrUTP showed that peroxisomes were the sites of nascent viral synthesis, whereas in situ hybridization indicated that DI RNA progeny were diffused throughout the cytoplasm. DI RNA replication also took place in yeast cells devoid of peroxisomes. It is suggested that replication in these cells was targeted to the endoplasmic reticulum. PMID- 16476995 TI - Essential role of the Box II cis element and cognate host factors in regulating the promoter of Rice tungro bacilliform virus. AB - Rice tungro bacilliform virus (RTBV) is a double-stranded DNA virus with a single, tissue-specific promoter that is expressed primarily in phloem tissues. Rice transcription factors RF2a and RF2b bind to Box II, a cis element adjacent to the TATA box, and control gene expression from the promoter. Mutations were made in the promoter to delete or mutate Box II and the mutated promoters were fused to a reporter gene; the chimeric genes were expressed in transient BY-2 protoplast assays and in transgenic Arabidopsis plants. The results of these studies showed that Box II is essential to the activity of the RTBV promoter. A chimeric beta-glucuronidase (GUS) reporter gene containing the Box II sequence and a minimal promoter derived from the Cauliflower mosaic virus 35S promoter were co-transfected into protoplasts with gene constructs that encoded RF2a or RF2b. The reporter gene produced threefold higher GUS activity when co transfected with RF2a, and 11-fold higher activity when co-transfected with RF2b, than in the absence of added transcription factors. Moreover, chimeric reporter genes were activated by approximately threefold following induction of expression of the RF2a gene in transgenic Arabidopsis plants. The work presented here and earlier findings show that Box II and its interactions with cognate rice transcription factors, including RF2a and RF2b, are essential to the activity of the RTBV promoter and are probably involved in expression of the RTBV genome during virus replication. PMID- 16476996 TI - Complete nucleotide sequence and genome organization of a single-stranded RNA virus infecting the marine fungoid protist Schizochytrium sp. AB - The complete nucleotide sequence of the genomic RNA of a marine fungoid protist infecting virus (Schizochytrium single-stranded RNA virus; SssRNAV) has been determined. The viral RNA is single-stranded with a positive sense and is 9,018 nt in length [excluding the 3' poly(A) tail]. It contains two long open reading frames (ORFs), which are separated by an intergenic region of 92 nt. The 5' ORF (ORF1) is preceded by an untranslated leader sequence of 554 nt. The 3' large ORF (ORF2) and an additional ORF (ORF3) overlap ORF2 by 431 nt and are followed by an untranslated region of 70 nt [excluding the 3' poly(A) tail]. The deduced amino acid sequences of ORF1 and ORF2 products show similarity to non-structural and structural proteins of dicistroviruses, respectively. However, Northern blot analysis suggests that SssRNAV synthesizes subgenomic RNAs to translate ORF2 and ORF3, showing that the translation mechanism of downstream ORFs is distinct from that of dicistroviruses. Furthermore, although considerable similarities were detected by using a blast genome database search, phylogenetic analysis based on both the nucleotide and amino acid sequences of the putative RNA-dependent RNA polymerase (RdRp) and the RNA helicase suggests that SssRNAV is phylogenetically distinct from other virus families. Therefore, it is concluded that SssRNAV is not a member of any currently defined virus family and belongs to a novel, unrecognized virus group. PMID- 16476998 TI - The structure of a ring-opened proliferating cell nuclear antigen-replication factor C complex revealed by fluorescence energy transfer. AB - Numerous proteins that function in DNA metabolic pathways are known to interact with the proliferating cell nuclear antigen (PCNA). The important function of PCNA in stimulating various cellular activities requires its topological linkage with DNA. Loading of the circular PCNA onto duplex DNA requires the activity of a clamp-loader [replication factor C (RFC)] complex and the energy derived from ATP hydrolysis. The mechanistic and structural details regarding PCNA loading by the RFC complex are still developing. In particular, the positive identification of a long-hypothesized structure of an open clamp-RFC complex as an intermediate in loading has remained elusive. In this study, we capture an open yeast PCNA clamp in a complex with RFC through fluorescence energy transfer experiments. We also follow the topological transitions of PCNA in the various steps of the clamp loading pathway through both steady-state and stopped-flow fluorescence studies. We find that ATP effectively drives the clamp-loading process to completion with the formation of the closed PCNA bound to DNA, whereas ATPgammaS cannot. The information derived from this work complements that obtained from previous structural and mechanistic studies and provides a more complete picture of a eukaryotic clamp-loading pathway using yeast as a paradigm. PMID- 16476997 TI - Role for calcium/calmodulin-dependent protein kinase II in the p75-mediated regulation of sympathetic cholinergic transmission. AB - Neurotrophins regulate sympathetic neuron cotransmission by modulating the activity-dependent release of norepinephrine and acetylcholine. Nerve growth factor promotes excitatory noradrenergic transmission, whereas brain-derived neurotrophic factor (BDNF), acting through the p75 receptor, increases inhibitory cholinergic transmission. This regulation of corelease by target-derived factors leads to the functional modulation of myocyte beat rate in neuron-myocyte cocultures. Calcium/calmodulin-dependent protein kinase II (CaMKII) has been implicated in the control of both pre- and postsynaptic mechanisms of synaptic plasticity. We demonstrate that CaMKII acts in conjunction with p75 signaling to regulate cholinergic transmission between sympathetic neurons and heart cells. Inhibition of presynaptic CaMKII prevents the BDNF-dependent shift to inhibitory neurotransmission, whereas presynaptic expression of a constitutively active CaMKII results in inhibitory neurotransmission in the absence of added BDNF, suggesting that activation of presynaptic CaMKII is both necessary and sufficient for a shift from excitatory to inhibitory transmission. Several isozymes of CaMKII are expressed in sympathetic neurons, with the delta-CaMKII being activated by BDNF and nerve growth factor. Activated CaMKII is less effective at promoting cholinergic transmission in the absence of p75 signaling, demonstrating that p75 and CaMKII act to coordinate neurotransmitter selection in sympathetic neurons. PMID- 16477000 TI - DNA conformation on surfaces measured by fluorescence self-interference. AB - The conformation of DNA molecules tethered to the surface of a microarray may significantly affect the efficiency of hybridization. Although a number of methods have been applied to determine the structure of the DNA layer, they are not very sensitive to variations in the shape of DNA molecules. Here we describe the application of an interferometric technique called spectral self-interference fluorescence microscopy to the precise measurement of the average location of a fluorescent label in a DNA layer relative to the surface and thus determine specific information on the conformation of the surface-bound DNA molecules. Using spectral self-interference fluorescence microscopy, we have estimated the shape of coiled single-stranded DNA, the average tilt of double-stranded DNA of different lengths, and the amount of hybridization. The data provide important proofs of concept for the capabilities of novel optical surface analytical methods of the molecular disposition of DNA on surfaces. The determination of DNA conformations on surfaces and hybridization behavior provide information required to move DNA interfacial applications forward and thus impact emerging clinical and biotechnological fields. PMID- 16476999 TI - Peptides selected for binding to clotted plasma accumulate in tumor stroma and wounds. AB - Screening of a phage library for peptides that bind to clotted plasma in the presence of liquid plasma yielded two cyclic decapeptides, CGLIIQKNEC (CLT1) and CNAGESSKNC (CLT2). When injected intravenously into mice bearing various types of tumors, fluorescein-conjugated CLT peptides accumulated in a fibrillar meshwork in the extracellular compartment of the tumors, but were not detectable in other tissues of the tumor-bearing mice. The tumor homing of both peptides was strongly reduced after coinjection with unlabeled CLT2, indicating that the two peptides recognize the same binding site. The CLT peptide fluorescence colocalized with staining for fibrin(ogen) present in the extravascular compartment of tumors, but not in other tissues. The CLT peptides did not home to tumors grown in fibrinogen null mice or in mice that lack plasma fibronectin. The CLT peptides also accumulated at the sites of injury in arteries, skeletal muscle, and skin. We conclude that the CLT peptides recognize fibrin-fibronectin complexes formed by clotting of plasma proteins that have leaked into the extravascular space in tumors and other lesions. These peptides may be useful in targeting diagnostic and therapeutic materials into tumors and injured tissues. PMID- 16477001 TI - Activated Src abrogates the Myc requirement for the G0/G1 transition but not for the G1/S transition. AB - The transcription factor Myc plays a central role in the control of cellular proliferation. Myc expression is induced by growth factors in a pathway mediated by cellular Src (c-Src), but it is not clear whether Myc induction or activity is required for malignant transformation by activated Src. We introduced v-Src into a c-myc(-/-) derivative of Rat-1 fibroblasts and into 3T9 mouse fibroblasts harboring a conditionally excisable c-myc allele. Expression of activated viral Src in Myc-deficient cells led to loss of actin stress fibers and surface fibronectin, indicating that Myc is dispensable for v-Src-induced morphological transformation. However, v-Src failed to rescue the proliferative defect resulting from the loss of Myc. In Myc-deficient cells, despite its inability to overcome this proliferation block, v-Src was able to regulate the expression of certain Myc transcriptional targets and induce the expression of active cyclin D/Cdk4 and Cdk6 complexes; it also induced the phosphorylation of Rb, albeit at reduced levels. In contrast, however, in the absence of Myc, the level of Cdk2 kinase activity was drastically reduced. This reduction in Cdk2 activity was associated with a decrease in the expression of Cdk7, Cdc25A, and cyclin A. Coexpression of Cdk2 plus cyclin E and/or cyclin A rescued the G1/S block and allowed the cells to enter mitosis. These results indicate that in the absence of Myc, v-Src can activate early G1 cell cycle regulators but fails to activate regulators of the late G1/S transition. PMID- 16477002 TI - Crystal structure of the IL-2 signaling complex: paradigm for a heterotrimeric cytokine receptor. AB - IL-2 is a cytokine that functions as a growth factor and central regulator in the immune system and mediates its effects through ligand-induced hetero trimerization of the receptor subunits IL-2R alpha, IL-2R beta, and gamma(c). Here, we describe the crystal structure of the trimeric assembly of the human IL 2 receptor ectodomains in complex with IL-2 at 3.0 A resolution. The quaternary structure is consistent with a stepwise assembly from IL-2/IL-2R alpha to IL-2/IL 2R alpha/IL-2R beta to IL-2/IL-2R alpha/IL-2R beta/gamma(c). The IL-2R alpha subunit forms the largest of the three IL-2/IL-2R interfaces, which, together with the high abundance of charge-charge interactions, correlates well with the rapid association rate and high-affinity interaction of IL-2R alpha with IL-2 at the cell surface. Surprisingly, IL-2R alpha makes no contacts with IL-2R beta or gamma(c), and only minor changes are observed in the IL-2 structure in response to receptor binding. These findings support the principal role of IL-2R alpha to deliver IL-2 to the signaling complex and act as regulator of signal transduction. Cooperativity in assembly of the final quaternary complex is easily explained by the extraordinarily extensive set of interfaces found within the fully assembled IL-2 signaling complex, which nearly span the entire length of the IL-2R beta and gamma(c) subunits. Helix A of IL-2 wedges tightly between IL 2R beta and gamma(c) to form a three-way junction that coalesces into a composite binding site for the final gamma(c) recruitment. The IL-2/gamma(c) interface itself exhibits the smallest buried surface and the fewest hydrogen bonds in the complex, which is consistent with its promiscuous use in other cytokine receptor complexes. PMID- 16477003 TI - Kappa opioids selectively control dopaminergic neurons projecting to the prefrontal cortex. AB - Dopaminergic afferents arising from the ventral tegmental area (VTA) are crucial elements in the neural circuits that mediate arousal, motivation, and reinforcement. Two major targets of these afferents are the medial prefrontal cortex (mPFC) and the nucleus accumbens (NAc). Whereas dopamine (DA) in the mPFC has been implicated in working memory and attentional processes, DA in the NAc is required for responding to reward predictive cues. These distinct functions suggest a role for independent firing patterns of dopaminergic neurons projecting to these brain regions. In fact, DA release in mPFC and NAc can be differentially modulated. However, to date, electrophysiological studies have largely overlooked heterogeneity among VTA neurons. Here, we provide direct evidence for differential neurotransmitter control of DA neural activity and corresponding DA release based on projection target. Kappa opioid receptor agonists inhibit VTA DA neurons that project to the mPFC but not those that project to the NAc. Moreover, DA levels in the mPFC, but not the NAc, are reduced after local infusion of kappa opioid receptor agonists into the VTA. These findings demonstrate that DA release in specific brain regions can be independently regulated by opioid targeting of a subpopulation of VTA DA neurons. Selective control of VTA DA neurons projecting to the mPFC has important implications for understanding addiction, attention disorders, and schizophrenia, all of which are associated with DA dysfunction in the mPFC. PMID- 16477004 TI - An enzymatic ruler modulates Lewis antigen glycosylation of Helicobacter pylori LPS during persistent infection. AB - Helicobacter pylori persistently colonizes about half the human population and contributes to the development of peptic ulcer disease and gastric cancer. This organism has evolved means to structurally alter its surface characteristics to evade innate and adaptive immune responses. H. pylori produces LPS O-antigen units that can be posttranslationally fucosylated to generate Lewis antigens, structures also found on human epithelial cells. We demonstrate an extensive diversity of Lewis x and Lewis y expression in LPS O-antigen units, occurring over time and in different regions of the human stomach. Lewis expression patterns were correlated with the on/off status of the three fucosyltransferases (FucT), FutA, FutB, and FutC, which are regulated via slipped-strand mispairing in intragenic polyC tract regions of the corresponding genes. The alpha1,3-FucT, FutA and FutB, each contain a C-terminal heptad repeat region, consisting of a variable number of DD/NLRV/INY tandem repeats. Variations in the number of heptad repeats correlated to the sizes of O-antigen polymers to become decorated by fucose residues. Our data support a molecular ruler mechanism for how H. pylori varies its LPS fucosylation pattern, where one heptad repeat in the enzyme corresponds to one N-acetyl-beta-lactosamine unit in the O-antigen polysaccharide. PMID- 16477005 TI - An ordered, nonredundant library of Pseudomonas aeruginosa strain PA14 transposon insertion mutants. AB - Random transposon insertion libraries have proven invaluable in studying bacterial genomes. Libraries that approach saturation must be large, with multiple insertions per gene, making comprehensive genome-wide scanning difficult. To facilitate genome-scale study of the opportunistic human pathogen Pseudomonas aeruginosa strain PA14, we constructed a nonredundant library of PA14 transposon mutants (the PA14NR Set) in which nonessential PA14 genes are represented by a single transposon insertion chosen from a comprehensive library of insertion mutants. The parental library of PA14 transposon insertion mutants was generated by using MAR2xT7, a transposon compatible with transposon-site hybridization and based on mariner. The transposon-site hybridization genetic footprinting feature broadens the utility of the library by allowing pooled MAR2xT7 mutants to be individually tracked under different experimental conditions. A public, internet-accessible database (the PA14 Transposon Insertion Mutant Database, http://ausubellab.mgh.harvard.edu/cgi-bin/pa14/home.cgi) was developed to facilitate construction, distribution, and use of the PA14NR Set. The usefulness of the PA14NR Set in genome-wide scanning for phenotypic mutants was validated in a screen for attachment to abiotic surfaces. Comparison of the genes disrupted in the PA14 transposon insertion library with an independently constructed insertion library in P. aeruginosa strain PAO1 provides an estimate of the number of P. aeruginosa essential genes. PMID- 16477006 TI - IL-1 receptor-associated kinase 1 is critical for latent membrane protein 1 induced p65/RelA serine 536 phosphorylation and NF-kappaB activation. AB - Epstein-Barr virus latent infection integral membrane protein 1 (LMP1) mimics a constitutively active TNF receptor (TNFR). LMP1 has two C-terminal cytosolic domains, transformation effector sites (TES)1 and -2, that engage TNFR-associated factors (TRAFs) and the TNFR-associated death domain protein, respectively, and activate NF-kappaB. NF-kappaB activation is critical for Epstein-Barr virus infected lymphoblast survival. TES1- and TES2-mediated NF-kappaB activations are IL-1 receptor-associated kinase 1 (IRAK1)-dependent. Because IRAK1 is upstream of TRAF6 in IL-1 activation of NF-kappaB, the potential role of IRAK1 in LMP1 mediated NF-kappaB activation through TRAF6 and inhibitor of kappaB (IkappaB) kinase (IKK) was initially investigated. Surprisingly, LMP1 expression activated TRAF6 ubiquitination, IKKbeta induction of IkappaB alpha phosphorylation, and p65 nuclear translocation in both WT and IRAK1-deficient I1A 293 cells. LMP1 also induced IKK alpha-mediated p100 processing and p52 nuclear localization in WT and IRAK1-deficient I1A 293 cells. Further, LMP1 TES1 and TES2 induced p65, p50, and p52 NF-kappaB DNA binding in WT and IRAK1-deficient I1A 293 cells. However, LMP1 induced p65/RelA S536 phosphorylation only in WT 293 cells or in IRAK1 kinase point mutant reconstituted I1A 293 cells but not in IRAK1-deficient I1A 293 cells. IRAK1 was also required for LMP1 activation of p38, one of the kinases that can mediate p65/RelA S536 phosphorylation and activate NF-kappaB-dependent transcription. Thus, the critical IRAK1 role in LMP1-induced NF-kappaB activation is in mediating p65/RelA S536 phosphorylation through an effect on p38 or other p65 S536 kinases. PMID- 16477007 TI - Analysis of Pseudomonas aeruginosa diguanylate cyclases and phosphodiesterases reveals a role for bis-(3'-5')-cyclic-GMP in virulence. AB - The opportunistic pathogen Pseudomonas aeruginosa is responsible for systemic infections in immunocompromised individuals and chronic respiratory disease in patients with cystic fibrosis. Cyclic nucleotides are known to play a variety of roles in the regulation of virulence-related factors in pathogenic bacteria. A set of P. aeruginosa genes, encoding proteins that contain putative domains characteristic of diguanylate cyclases (DGCs) and phosphodiesterases (PDEs) that are responsible for the maintenance of cellular levels of the second messenger bis-(3'-5')-cyclic dimeric GMP (c-di-GMP) was identified in the annotated genomes of P. aeruginosa strains PAO1 and PA14. Although the majority of these genes are components of the P. aeruginosa core genome, several are located on presumptive horizontally acquired genomic islands. A comprehensive analysis of P. aeruginosa genes encoding the enzymes of c-di-GMP metabolism (DGC- and PDE-encoding genes) was carried out to analyze the function of c-di-GMP in two disease-related phenomena, cytotoxicity and biofilm formation. Analysis of the phenotypes of DGC and PDE mutants and overexpressing clones revealed that certain virulence associated traits are controlled by multiple DGCs and PDEs through alterations in c-di-GMP levels. A set of mutants in selected DGC- and PDE-encoding genes exhibited attenuated virulence in a mouse infection model. Given that insertions in different DGC and PDE genes result in distinct phenotypes, it seems likely that the formation or degradation of c-di-GMP by these enzymes is in highly localized and intimately linked to particular targets of c-di-GMP action. PMID- 16477008 TI - Phanerozoic marine biodiversity dynamics in light of the incompleteness of the fossil record. AB - Long-term evolutionary dynamics have been approached through quantitative analysis of the fossil record, but without explicitly taking its incompleteness into account. Here we explore the temporal covariance structure of per-genus origination and extinction rates for global marine fossil genera throughout the Phanerozoic, both before and after corrections for the incompleteness of the fossil record. Using uncorrected data based on Sepkoski's compendium, we find significant autocovariance within origination and extinction rates, as well as covariance between extinction and origination, not one, but two, intervals later, corroborating evidence for the unexplained temporal gap found by past studies. However, these effects vanish when the data are corrected for the incompleteness of the fossil record. Instead, we observe significant covariance only between extinction and origination in the immediately following intervals. The gap in the response of the biosphere to extinction in the uncorrected fossil record thus appears to be an artifact of the incompleteness of the fossil record, specifically due to episodic variation in the probability that taxa will be preserved, on time scales comparable to the temporal resolution of Sepkoski's data. Our results also indicate that at that temporal resolution (the stage/substage of duration approximately = 5 million years), changes in origination and extinction do not persist for longer than one interval, except that elevated origination rates immediately after extinction may last for more than a single interval. Thus, although certain individual cases may deviate from the overall pattern, we find that in general the biosphere's response to perturbation is immediate geologically and usually short-lived. PMID- 16477009 TI - NMDA receptor mediates tau-induced neurotoxicity by calpain and ERK/MAPK activation. AB - The altered function and/or structure of tau protein is postulated to cause cell death in tauopathies and Alzheimer's disease. However, the mechanisms by which tau induces neuronal death remain unclear. Here we show that overexpression of human tau and of some of its N-terminal fragments in primary neuronal cultures leads to an N-methyl-D-aspartate receptor (NMDAR)-mediated and caspase independent cell death. Death signaling likely originates from stimulation of extrasynaptic NR2B-subunit-containing NMDARs because it is accompanied by dephosphorylation of cAMP-response-element-binding protein (CREB) and it is inhibited by ifenprodil. Interestingly, activation of NMDAR leads to a crucial, sustained, and delayed phosphorylation of extracellular-regulated kinases 1 and 2, whose inhibition largely prevents tau-induced neuronal death. Moreover, NMDAR involvement causes the fatal activation of calpain, which, in turn, degrades tau protein into a 17-kDa peptide and possibly other highly toxic N-terminal peptides. Some of these peptides are hypothesized, on the basis of our in vitro experiments, to initiate a negative loop, ultimately leading to cell death. Thus, inhibition of calpain largely prevents tau degradation and cell death. Our findings unravel a cellular mechanism linking tau toxicity to NMDAR activation and might be relevant to Alzheimer's disease and tauopathies where NMDAR-mediated toxicity is postulated to play a pivotal role. PMID- 16477010 TI - Cell-type-specific signatures of microRNAs on target mRNA expression. AB - Although it is known that the human genome contains hundreds of microRNA (miRNA) genes and that each miRNA can regulate a large number of mRNA targets, the overall effect of miRNAs on mRNA tissue profiles has not been systematically elucidated. Here, we show that predicted human mRNA targets of several highly tissue-specific miRNAs are typically expressed in the same tissue as the miRNA but at significantly lower levels than in tissues where the miRNA is not present. Conversely, highly expressed genes are often enriched in mRNAs that do not have the recognition motifs for the miRNAs expressed in these tissues. Together, our data support the hypothesis that miRNA expression broadly contributes to tissue specificity of mRNA expression in many human tissues. Based on these insights, we apply a computational tool to directly correlate 3' UTR motifs with changes in mRNA levels upon miRNA overexpression or knockdown. We show that this tool can identify functionally important 3' UTR motifs without cross-species comparison. PMID- 16477011 TI - Biogeographical distribution and diversity of microbes in methane hydrate-bearing deep marine sediments on the Pacific Ocean Margin. AB - The deep subseafloor biosphere is among the least-understood habitats on Earth, even though the huge microbial biomass therein plays an important role for potential long-term controls on global biogeochemical cycles. We report here the vertical and geographical distribution of microbes and their phylogenetic diversities in deeply buried marine sediments of the Pacific Ocean Margins. During the Ocean Drilling Program Legs 201 and 204, we obtained sediment cores from the Peru and Cascadia Margins that varied with respect to the presence of dissolved methane and methane hydrate. To examine differences in prokaryotic distribution patterns in sediments with or without methane hydrates, we studied >2,800 clones possessing partial sequences (400-500 bp) of the 16S rRNA gene and 348 representative clone sequences (approximately 1 kbp) from the two geographically separated subseafloor environments. Archaea of the uncultivated Deep-Sea Archaeal Group were consistently the dominant phylotype in sediments associated with methane hydrate. Sediment cores lacking methane hydrates displayed few or no Deep-Sea Archaeal Group phylotypes. Bacterial communities in the methane hydrate-bearing sediments were dominated by members of the JS1 group, Planctomycetes, and Chloroflexi. Results from cluster and principal component analyses, which include previously reported data from the West and East Pacific Margins, suggest that, for these locations in the Pacific Ocean, prokaryotic communities from methane hydrate-bearing sediment cores are distinct from those in hydrate-free cores. The recognition of which microbial groups prevail under distinctive subseafloor environments is a significant step toward determining the role these communities play in Earth's essential biogeochemical processes. PMID- 16477013 TI - Negative regulation of activation-induced cytidine deaminase in B cells. AB - Both class switch recombination (CSR) and somatic hypermutation (SHM) of the Ig genes require the activity of activation-induced cytidine deaminase (AID). Expression of AID is restricted to B cells in the germinal centers of the lymphoid organs, where activated B cells undergo CSR and SHM. We previously showed that constitutive and systemic expression of AID leads to tumorigenesis in T cells and lung epithelium, but not in B cells. This finding led us to suspect that transgenic AID may be inactivated at least in part in B cells. To address this issue, we generated conditional AID-transgenic mice that constitutively express AID only in B cells. Studies on the cross between the AID-transgenic and AID-deficient mice showed that abundant AID protein accumulated by constitutive expression is inactivated in B cells, possibly providing an explanation for the absence of deregulation of CSR and SHM in AID-transgenic B cells. PMID- 16477012 TI - Synergistic roles of platelet-derived growth factor-BB and interleukin-1beta in phenotypic modulation of human aortic smooth muscle cells. AB - The phenotype of smooth muscle cells (SMCs) plays an important role in vascular function in health and disease. We investigated the mechanism of modulation of SMC phenotype (from contractile to synthetic) induced by the synergistic action of a growth factor (platelet-derived growth factor, PDGF-BB) and a cytokine (interleukin, IL-1beta). Human aortic SMCs grown on polymerized collagen showed high expression levels of contractile markers (smooth muscle alpha-actin, myosin heavy chain, and calponin). These levels were not significantly affected by PDGF BB and IL-1beta individually, but decreased markedly after the combined usage of PDGF-BB and IL-1beta. PDGF/IL-1beta costimulation also induced a sustained phosphorylation of Akt and p70 ribosomal S6 kinase (p70S6K). The effects of PDGF/IL-1beta costimulation on contractile marker expression and Akt and p70S6K phosphorylation were blocked by the phosphatidylinositol 3-kinase inhibitors wortmannin and LY294002 and by adenovirus expressing a dominant-negative Akt, and they were mimicked by constitutively active Akt. PDGF-BB/IL-1beta induced a sustained phosphorylation of PDGF receptor (PDGFR)-beta and its association with IL-1 receptor (IL-1R1). Such activation and association of receptors were blocked by a PDGFR-beta neutralizing antibody (AF385), an IL-1R1 antagonist (IL-1ra), as well as a specific inhibitor of PDGFR-beta phosphorylation (AG1295); these agents also eliminated the PDGF-BB/IL-1beta-induced signaling and phenotypic modulation. PDGF-BB/IL-1beta inhibited the polymerized collagen-induced serum response factor DNA binding activity in the nucleus, and this effect was mediated by the PDGFR beta/IL-1R1 association and phosphatidylinositol 3-kinase/Akt/p70S6K pathway. Our findings provide insights into the mechanism of SMC phenotypic modulation from contractile to synthetic, e.g., in atherosclerosis. PMID- 16477014 TI - Ceramide mediates the rapid phase of febrile response to IL-1beta. AB - IL-1beta was identified after a long search for the endogenous pyrogen. It acts by inducing synthesis of prostaglandin E2, which mediates the late phase of IL 1beta-induced fever. Here we show by radiotelemetry that the early phase of the fever response to IL-1beta is mediated by ceramide. Hypothalamic application of the cell-penetrating C2-ceramide mimics the rapid phase of the IL-1beta-induced fever. Inhibition of ceramide synthesis blocks the rapid phase of fever but does not affect the slower prostaglandin E2-dependent phase, which is blocked by indomethacin or by null mutation of the EP3 prostanoid receptor. Electrophysiological experiments on preoptic area/anterior hypothalamic neurons show that C2-ceramide, but not dihydroceramide, mimics the rapid hyperpolarizing effects of IL-1beta on the activity of warm-sensitive hypothalamic neurons. IL 1beta-mediated hyperpolarization is blocked by PP2, the selective inhibitor of the protein tyrosine kinase Src, which is known to be activated by ceramide. These in vivo and in vitro data suggest that ceramide fulfills the criteria for an endogenous pyrogen. PMID- 16477015 TI - Multiconformation continuum electrostatics analysis of the NhaA Na+/H+ antiporter of Escherichia coli with functional implications. AB - Sodium proton antiporters are essential enzymes that catalyze the exchange of sodium ions for protons across biological membranes. Protonations and deprotonations of individual amino acid residues and of clusters formed by these residues play an important role in activating these enzymes and in the mechanism of transport. We have used multiconformation continuum electrostatics method to investigate the protonation states of residues in the sodium proton exchanger NhaA from Escherichia coli, the structure of which has been determined recently by x-ray crystallography. Our calculations identify four clusters of electrostatically tightly interacting residues as well as long-range interactions between residues required for activation. The importance of many of these residues has been demonstrated by the characterization of site-directed mutants. A number of residues with extreme pKa values, including several of the "pH sensor," can only undergo protonation/deprotonation reactions subsequent to conformational changes. The results of the calculations provide valuable information on the activation of the antiporter and the role of individual amino acid residues, and provide a solid framework for further experiments. PMID- 16477016 TI - Directly observed reversible shape changes and hemoglobin stratification during centrifugation of human and Amphiuma red blood cells. AB - This paper describes changes that occur in human and Amphiuma red blood cells observed during centrifugation with a special microscope. Dilute suspensions of cells were layered, in a centrifuge chamber, above an osmotically matched dense solution, containing Nycodenz, Ficoll, or Percoll (Pharmacia) that formed a density gradient that allowed the cells to slowly settle to an equilibrium position. Biconcave human red blood cells moved downward at low forces with minimum wobble. The cells oriented vertically when the force field was increased and Hb sedimented as the lower part of each cell became bulged and assumed a "bag like" shape. The upper centripetal portion of the cell became thinner and remained biconcave. These changes occurred rapidly and were completely reversible upon lowering the centrifugal force. Bag-shaped cells, upon touching red cells in rouleau, immediately reverted to biconcave disks as they flipped onto a stack. Amphiuma red cells displayed a different type of reversible stratification and deformation at high force fields. Here the cells became stretched, with the nucleus now moving centrifugally, the Hb moving centripetally, and the bottom of the cells becoming thinner and clear. Nevertheless, the distribution of the marginal bands at the cells' rim was unchanged. We conclude that centrifugation, per se, while changing a red cell's shape and the distribution of its intracellular constituents, does so in a completely reversible manner. Centrifugation of red cells harboring altered or missing structural elements could provide information on shape determinants that are still unexplained. PMID- 16477017 TI - Activation of IFN pathways and plasmacytoid dendritic cell recruitment in target organs of primary Sjogren's syndrome. AB - Gene expression analysis of target organs might help provide new insights into the pathogenesis of autoimmune diseases. We used global gene expression profiling of minor salivary glands to identify patterns of gene expression in patients with primary Sjogren's syndrome (pSS), a common and prototypic systemic autoimmune disease. Gene expression analysis allowed for differentiating most patients with pSS from controls. The expression of 23 genes in the IFN pathways, including two Toll-like receptors (TLR8 and TLR9), was significantly different between patients and controls. Furthermore, the increased expression of IFN-inducible genes, BAFF and IFN-induced transmembrane protein 1, was also demonstrated in ocular epithelial cells by quantitative RT-PCR. In vitro activation showed that these genes were effectively modulated by IFNs in salivary gland epithelial cells, the target cells of autoimmunity in pSS. The activation of IFN pathways led us to investigate whether plasmacytoid dendritic cells were recruited in salivary glands. These IFN-producing cells were detected by immunohistochemistry in all patients with pSS, whereas none was observed in controls. In conclusion, our results support the pathogenic interaction between the innate and adaptive immune system in pSS. The persistence of the IFN signature might be related to a vicious circle, in which the environment interacts with genetic factors to drive the stimulation of salivary TLRs. PMID- 16477018 TI - Association of brefeldin A-inhibited guanine nucleotide-exchange protein 2 (BIG2) with recycling endosomes during transferrin uptake. AB - ADP-ribosylation factors (ARFs) are critical in vesicular trafficking. Brefeldin A-inhibited guanine nucleotide-exchange protein (BIG)1 and BIG2 activate ARFs by accelerating replacement of bound GDP with GTP. Additional and differing functions of these approximately 200-kDa proteins are now being recognized, as are their independent intracellular movements. Here, we describe the localization in COS7 cells by immunofluorescence microscopy of BIG2, but not BIG1, with structures that have characteristics of recycling endosomes during transferrin (Tfn) uptake and Tfn receptor (TfnR) recycling. Cell content of BIG2 and Rab11, but not TfnR, BIG1, Rab4, or Exo70, was increased after 60 min of Tfn uptake. BIG2, but not BIG1, appeared in density-gradient fractions containing TfnR, Rab11, and Exo70 after 60 min of Tfn uptake. Treatment of cells with BIG2 small interfering RNA (siRNA), but not BIG1 or control siRNAs, decreased BIG2 protein >90% without affecting BIG1, ARF, or actin content, whereas TfnR was significantly increased as was its accumulation in perinuclear recycling endosomes. Tfn release appeared unaffected by BIG1 siRNA but was significantly slowed from cells treated with BIG2 siRNA alone or plus BIG1 siRNA. We suggest that BIG2 has an important role in Tfn uptake and TfnR recycling, perhaps through its demonstrated interaction with Exo70 and the exocyst complex. PMID- 16477020 TI - Structure and mechanism of the Propionibacterium acnes polyunsaturated fatty acid isomerase. AB - Conjugated linoleic acids (CLAs) affect body fat gain, carcinogenesis, insulin resistance, and lipid peroxidation in mammals. Several isomers of CLA exist, of which the (9Z, 11E) and (10E, 12Z) isomers have beneficial effects on human metabolism but are scarce in foods. Bacterial polyunsaturated fatty acid isomerases are promising biotechnological catalysts for CLA production. We describe six crystal structures of the Propionibacterium acnes polyunsaturated fatty acid isomerase PAI in apo- and product-bound forms. The three-domain flavoprotein has previously undescribed folds outside the FAD-binding site. Conformational changes in a hydrophobic channel toward the active site reveal a unique gating mechanism for substrate specificity. The geometry of the substrate binding site explains the length preferences for C18 fatty acids. A catalytic mechanism for double-bond isomerization is formulated that may be altered to change substrate specificity for syntheses of rare CLAs from easily accessible precursors. PMID- 16477019 TI - Gene expression changes associated with progression and response in chronic myeloid leukemia. AB - Chronic myeloid leukemia (CML) is a hematopoietic stem cell disease with distinct biological and clinical features. The biologic basis of the stereotypical progression from chronic phase through accelerated phase to blast crisis is poorly understood. We used DNA microarrays to compare gene expression in 91 cases of CML in chronic (42 cases), accelerated (17 cases), and blast phases (32 cases). Three thousand genes were found to be significantly (P < 10(-10)) associated with phase of disease. A comparison of the gene signatures of chronic, accelerated, and blast phases suggest that the progression of chronic phase CML to advanced phase (accelerated and blast crisis) CML is a two-step rather than a three-step process, with new gene expression changes occurring early in accelerated phase before the accumulation of increased numbers of leukemia blast cells. Especially noteworthy and potentially significant in the progression program were the deregulation of the WNT/beta-catenin pathway, the decreased expression of Jun B and Fos, alternative kinase deregulation, such as Arg (Abl2), and an increased expression of PRAME. Studies of CML patients who relapsed after initially successful treatment with imatinib demonstrated a gene expression pattern closely related to advanced phase disease. These studies point to specific gene pathways that might be exploited for both prognostic indicators as well as new targets for therapy. PMID- 16477021 TI - CSPalpha-deficiency causes massive and rapid photoreceptor degeneration. AB - Cysteine string protein (CSP) alpha is an abundant synaptic vesicle protein that contains a DNA-J domain characteristic of Hsp40-type cochaperones. Previous studies showed that deletion of CSPalpha in mice leads to massive lethal neurodegeneration but did not clarify how the neurodegeneration affects specific subpopulations of neurons. Here, we analyzed the effects of the CSPalpha deficiency on tonically active ribbon synapses of the retina and the inner ear. We show that CSPalpha-deficient photoreceptor terminals undergo dramatic and rapidly progressive neurodegeneration that starts before eye opening and initially does not affect other retinal synapses. These changes are associated with progressive blindness. In contrast, ribbon synapses of auditory hair cells did not exhibit presynaptic impairments in CSPalpha-deficient mice. Hair cells, but not photoreceptor cells or central neurons, express CSPbeta, thereby accounting for the lack of a hair-cell phenotype in CSPalpha knockout mice. Our data demonstrate that tonically active ribbon synapses in retina are particularly sensitive to the deletion of CSPalpha and that expression of at least one CSP isoform is essential to protect such tonically active synapses from neurodegeneration. PMID- 16477022 TI - SmyD1, a histone methyltransferase, is required for myofibril organization and muscle contraction in zebrafish embryos. AB - Histone modification has emerged as a fundamental mechanism for control of gene expression and cell differentiation. Recent studies suggest that SmyD1, a novo SET domain-containing protein, may play a critical role in cardiac muscle differentiation. However, its role in skeletal muscle development and its mechanism of actions remains elusive. Here we report that SmyD1a and SmyD1b, generated by alternative splicing of SmyD1 gene, are histone methyltransferases that play a key role in skeletal and cardiac muscle contraction. SmyD1a and SmyD1b are specifically expressed in skeletal and cardiac muscles of zebrafish embryos. Knockdown of SmyD1a and SmyD1b expression by morpholino antisense oligos resulted in malfunction of skeletal and cardiac muscles. The SmyD1 morphant embryos (embryos injected with morpholino oligos) could not swim and had no heartbeat. Myofibril organization in the morphant embryos was severely disrupted. The affected myofibers appeared as immature fibers with centrally located nuclei. Together, these data indicate that SmyD1a and SmyD1b are histone methyltransferases and play a critical role in myofibril organization during myofiber maturation. PMID- 16477023 TI - Crystal structure of human indoleamine 2,3-dioxygenase: catalytic mechanism of O2 incorporation by a heme-containing dioxygenase. AB - Human indoleamine 2,3-dioxygenase (IDO) catalyzes the cleavage of the pyrrol ring of L-Trp and incorporates both atoms of a molecule of oxygen (O2). Here we report on the x-ray crystal structure of human IDO, complexed with the ligand inhibitor 4-phenylimidazole and cyanide. The overall structure of IDO shows two alpha helical domains with the heme between them. A264 of the flexible loop in the heme distal side is in close proximity to the iron. A mutant analysis shows that none of the polar amino acid residues in the distal heme pocket are essential for activity, suggesting that, unlike the heme-containing monooxygenases (i.e., peroxidase and cytochrome P450), no protein group of IDO is essential in dioxygen activation or proton abstraction. These characteristics of the IDO structure provide support for a reaction mechanism involving the abstraction of a proton from the substrate by iron-bound dioxygen. Inactive mutants (F226A, F227A, and R231A) retain substrate-binding affinity, and an electron density map reveals that 2-(N-cyclohexylamino)ethane sulfonic acid is bound to these residues, mimicking the substrate. These findings suggest that strict shape complementarities between the indole ring of the substrate and the protein side chains are required, not for binding, but, rather, to permit the interaction between the substrate and iron-bound dioxygen in the first step of the reaction. This study provides the structural basis for a heme-containing dioxygenase mechanism, a missing piece in our understanding of heme chemistry. PMID- 16477024 TI - Protein tyrosine phosphatase 1B negatively regulates macrophage development through CSF-1 signaling. AB - Protein tyrosine phosphatase 1B (PTP-1B) is a ubiquitously expressed cytosolic phosphatase with the ability to dephosphorylate JAK2 and TYK2, and thereby down regulate cytokine receptor signaling. Furthermore, PTP-1B levels are up-regulated in certain chronic myelogenous leukemia patients, which points to a potential role for PTP-1B in myeloid development. The results presented here show that the absence of PTP-1B affects murine myelopoiesis by modifying the ratio of monocytes to granulocytes in vivo. This bias toward monocytic development is at least in part due to a decreased threshold of response to CSF-1, because the PTP-1B -/- bone marrow presents no abnormalities at the granulocyte-monocyte progenitor level but produces significantly more monocytic colonies in the presence of CSF 1. This phenomenon is not due to an increase in receptor levels but rather to enhanced phosphorylation of the activation loop tyrosine. PTP-1B -/- cells display increased inflammatory activity in vitro and in vivo through the constitutive up-regulation of activation markers as well as increased sensitivity to endotoxin. Collectively, our data indicate that PTP-1B is an important modulator of myeloid differentiation and macrophage activation in vivo and provide a demonstration of a physiological role for PTP-1B in immune regulation. PMID- 16477026 TI - Type III effector AvrPtoB requires intrinsic E3 ubiquitin ligase activity to suppress plant cell death and immunity. AB - Microbial pathogens of both plants and animals employ virulence factors that suppress the host immune response. The tomato pathogen Pseudomonas syringae injects the AvrPtoB type III effector protein into the plant cell to suppress programmed cell death (PCD) associated with plant immunity. AvrPtoB also inhibits PCD in yeast, indicating that AvrPtoB manipulates a conserved component of eukaryotic PCD. To identify host targets of AvrPtoB, we performed a yeast two hybrid screen and identified tomato ubiquitin (Ub) as a strong AvrPtoB interactor. AvrPtoB is ubiquitinated in vitro and exhibits E3 Ub ligase activity in the presence of recombinant E1 activating enzyme and specific E2 Ub conjugating enzymes. The C terminus of AvrPtoB is sufficient for both anti-PCD and E3 Ub ligase activities, suggesting the two functions are associated. Indeed, mutation of AvrPtoB lysine residues in the C terminus, between K512 and K529, disrupts AvrPtoB-Ub interactions, decreases AvrPtoB-mediated anti-PCD activity, and abrogates P. syringae pathogenesis of susceptible tomato plants. Remarkably, quantitative decreases in AvrPtoB anti-PCD activity are correlated with decreases in AvrPtoB ubiquitination and E3 Ub ligase activity. Overall, these data reveal a unique bacterial pathogenesis strategy, where AvrPtoB manipulates the host Ub system and requires intrinsic E3 Ub ligase activity to suppress plant immunity. PMID- 16477025 TI - Relevance and safety of telomerase for human tissue engineering. AB - Tissue engineering holds the promise of replacing damaged or diseased tissues and organs. The use of autologous donor cells is often not feasible because of the limited replicative lifespan of cells, particularly those derived from elderly patients. Proliferative arrest can be overcome by the ectopic expression of telomerase via human telomerase reverse transcriptase (hTERT) gene transfection. To study the efficacy and safety of this potentially valuable technology, we used differentiated vascular smooth muscle cells (SMC) and vascular tissue engineering as a model system. Although we previously demonstrated that vessels engineered with telomerase-expressing SMC had improved mechanics over those grown with control cells, it is critical to assess the phenotypic impact of telomerase expression in donor cells, because telomerase up-regulation is observed in >95% of human malignancies. To study the impact of telomerase in tissue engineering, expression of hTERT was retrovirally induced in SMC from eight elderly patients and one young donor. In hTERT SMC, significant lifespan extension beyond that of control was achieved without population doubling time acceleration. Karyotype changes were seen in both control and hTERT SMC but were not clonal nor representative of cancerous change. hTERT cells also failed to show evidence of neoplastic transformation in functional assays of tumorigenicity. In addition, the impact of donor age on cellular behavior, particularly the synthetic capability of SMC, was not affected by hTERT expression. Hence, this tissue engineering model system highlights the impact of donor age on cellular synthetic function that appears to be independent of lifespan extension by hTERT. PMID- 16477027 TI - Molecular structure of EmbR, a response element of Ser/Thr kinase signaling in Mycobacterium tuberculosis. AB - Ser/Thr phosphorylation has emerged as a critical regulatory mechanism in a number of bacteria, including Mycobacterium tuberculosis. This problematic pathogen encodes 11 eukaryotic-like Ser/Thr kinases, yet few substrates or signaling targets have been characterized. Here, we report the structure of EmbR (2.0 A), a putative transcriptional regulator of key arabinosyltransferases (EmbC, -A, and -B), and an endogenous substrate of the Ser/Thr-kinase PknH. EmbR presents a unique domain architecture: the N-terminal winged-helix DNA-binding domain forms an extensive interface with the all-helical central bacterial transcriptional activation domain and is positioned adjacent to the regulatory C terminal forkhead-associated (FHA) domain, which mediates binding to a Thr phosphorylated site in PknH. The structure in complex with a phospho-peptide (1.9 A) reveals a conserved mode of phospho-threonine recognition by the FHA domain and evidence for specific recognition of the cognate kinase. The present structures suggest hypotheses as to how EmbR might propagate the phospho-relay signal from its cognate kinase, while serving as a template for the structurally uncharacterized Streptomyces antibiotic regulatory protein family of transcription factors. PMID- 16477028 TI - Microscale technologies for tissue engineering and biology. AB - Microscale technologies are emerging as powerful tools for tissue engineering and biological studies. In this review, we present an overview of these technologies in various tissue engineering applications, such as for fabricating 3D microfabricated scaffolds, as templates for cell aggregate formation, or for fabricating materials in a spatially regulated manner. In addition, we give examples of the use of microscale technologies for controlling the cellular microenvironment in vitro and for performing high-throughput assays. The use of microfluidics, surface patterning, and patterned cocultures in regulating various aspects of cellular microenvironment is discussed, as well as the application of these technologies in directing cell fate and elucidating the underlying biology. Throughout this review, we will use specific examples where available and will provide trends and future directions in the field. PMID- 16477029 TI - Regulating activation of transplanted cells controls tissue regeneration. AB - Current approaches to tissue regeneration are limited by the death of most transplanted cells and/or resultant poor integration of transplanted cells with host tissue. We hypothesized that transplanting progenitor cells within synthetic microenvironments that maintain viability, prevent terminal differentiation, and promote outward migration would significantly enhance their repopulation and regeneration of damaged host tissue. This hypothesis was addressed in the context of muscle regeneration by transplanting satellite cells to muscle laceration sites on a delivery vehicle releasing factors that induce cell activation and migration (hepatocyte growth factor and fibroblast growth factor 2) or transplantation on materials lacking factor release. Controls included direct cell injection into muscle, the implantation of blank scaffolds, and scaffolds releasing factors without cells. Injected cells demonstrated a limited repopulation of damaged muscle and led to a slight improvement in muscle regeneration, as expected. Delivery of cells on scaffolds that did not promote migration resulted in no improvement in muscle regeneration. Strikingly, delivery of cells on scaffolds that promoted myoblast activation and migration led to extensive repopulation of host muscle tissue and increased the regeneration of muscle fibers at the wound and the mass of the injured muscle. This previously undescribed strategy for cell transplantation significantly enhances muscle regeneration from transplanted cells and may be broadly applicable to the various tissues and organ systems in which provision and instruction of a cell population competent to participate in regeneration may be clinically useful. PMID- 16477030 TI - A voltage-driven switch for ion-independent signaling by ether-a-go-go K+ channels. AB - Voltage-gated channels maintain cellular resting potentials and generate neuronal action potentials by regulating ion flux. Here, we show that Ether-a-go-go (EAG) K+ channels also regulate intracellular signaling pathways by a mechanism that is independent of ion flux and depends on the position of the voltage sensor. Regulation of intracellular signaling was initially inferred from changes in proliferation. Specifically, transfection of NIH 3T3 fibroblasts or C2C12 myoblasts with either wild-type or nonconducting (F456A) eag resulted in dramatic increases in cell density and BrdUrd incorporation over vector- and Shaker transfected controls. The effect of EAG was independent of serum and unaffected by changes in extracellular calcium. Inhibitors of p38 mitogen-activated protein (MAP) kinases, but not p44/42 MAP kinases (extracellular signal-regulated kinases), blocked the proliferation induced by nonconducting EAG in serum-free media, and EAG increased p38 MAP kinase activity. Importantly, mutations that increased the proportion of channels in the open state inhibited EAG-induced proliferation, and this effect could not be explained by changes in the surface expression of EAG. These results indicate that channel conformation is a switch for the signaling activity of EAG and suggest an alternative mechanism for linking channel activity to the activity of intracellular messengers, a role that previously has been ascribed only to channels that regulate calcium influx. PMID- 16477031 TI - Role of estrogen receptor beta in colonic epithelium. AB - Several papers report that the colon is one of the tissues regulated by estrogen receptor (ER)beta. To better understand the physiological role of ERbeta in colonic tissue, we have compared morphology, proliferation, and differentiation of colonic epithelium in ERbeta-/- mice and WT littermates. BrdUrd labeling revealed that the number of proliferating cells was higher in ERbeta-/- mice and that the migration of labeled cells toward the luminal surface was faster in ERbeta-/- mice than in WT littermates. Additionally, in the absence of ERbeta, there was a decrease in apoptosis, which was measured by immunohistochemical staining of cleaved caspase-3. The state of differentiation of the colonic epithelial cells was studied by using epithelial markers. In ERbeta-/- mice, there was a significant decrease in the expression of the differentiation marker cytokeratin (CK)20 and in the cellular adhesion molecules alpha-catenin (an adherens junction protein) and plectin (a hemidesmosomal protein). These changes were also evident by electron microscopy as abnormalities in tight junctions and in the number and shape of desmosomes in ERbeta-/- mice. These findings suggest a role for ERbeta in the organization and architectural maintenance of the colon. Furthermore, our results indicate that the rapidly proliferating cells of the colonic epithelium in ERbeta-/- mice are lost by increased shedding and not by increased apoptosis. In this way, hyperproliferative cells that lack ERbeta do not form hyperplastic lesions and do not accumulate in the superficial epithelium. PMID- 16477033 TI - A large family of ancient repeat elements in the human genome is under strong selection. AB - Although conserved noncoding elements (CNEs) constitute the majority of sequences under purifying selection in the human genome, they remain poorly understood. CNEs seem to be largely unique, with no large families of similar elements reported to date. Here, we search for CNEs among the ancestral repeat classes in the human genome and report the discovery of a large CNE family containing >900 members. This family belongs to the MER121 class of repeats. Although the MER121 family members show considerable sequence variation among one another, the individual copies show striking conservation in orthologous locations across the human, dog, mouse, and rat genomes. The element is also present and conserved in orthologous locations in the marsupial, but its genome-wide dispersal postdates the divergence from birds. The comparative genomic data indicate that MER121 does not encode a family of either protein-coding or RNA genes. Although the precise function of these elements remains unknown, the evidence suggests that this unusual family may play a cis-regulatory or structural role in mammalian genomes. PMID- 16477032 TI - Variable host-pathogen compatibility in Mycobacterium tuberculosis. AB - Mycobacterium tuberculosis remains a major cause of morbidity and mortality worldwide. Studies have reported human pathogens to have geographically structured population genetics, some of which have been linked to ancient human migrations. However, no study has addressed the potential evolutionary consequences of such longstanding human-pathogen associations. Here, we demonstrate that the global population structure of M. tuberculosis is defined by six phylogeographical lineages, each associated with specific, sympatric human populations. In an urban cosmopolitan environment, mycobacterial lineages were much more likely to spread in sympatric than in allopatric patient populations. Tuberculosis cases that did occur in allopatric hosts disproportionately involved high-risk individuals with impaired host resistance. These observations suggest that mycobacterial lineages are adapted to particular human populations. If confirmed, our findings have important implications for tuberculosis control and vaccine development. PMID- 16477034 TI - Liddle's syndrome mutations increase Na+ transport through dual effects on epithelial Na+ channel surface expression and proteolytic cleavage. AB - Liddle's syndrome, an inherited form of hypertension, is caused by mutations that delete or disrupt a C-terminal PY motif in the epithelial Na+ channel (ENaC). Previous work indicates that these mutations increase expression of ENaC at the cell surface by disrupting its binding to Nedd4-2, an E3 ubiquitin-protein ligase that targets ENaC for degradation. However, it remains uncertain whether this mechanism alone is responsible; increased activity of ENaC channels could also contribute to excessive Na+ transport in Liddle's syndrome. ENaC activity is controlled in part by its cleavage state; proteolytic cleavage produces channels with a high open-state probability, whereas uncleaved channels are inactive. Here, we found that Liddle's syndrome mutations have two distinct effects of ENaC surface expression, both of which contribute to increased Na+ transport. First, these mutations increased ENaC expression at the cell surface; second, they increased the fraction of ENaC at the cell surface that was cleaved (active). This disproportionate increase in cleavage was reproduced by expression of a dominant-negative Nedd4-2 or mutation of ENaC ubiquitination sites, interventions that disrupt ENaC endocytosis and lysosomal degradation. Conversely, overexpression of Nedd4-2 had the opposite effect, decreasing the fraction of cleaved ENaC at the cell surface. Thus, the data not only suggest that Nedd4-2 regulates epithelial Na+ transport in part by controlling the relative expression of cleaved and uncleaved ENaC at the cell surface but also provide a mechanism by which Liddle's syndrome mutations alter ENaC activity. PMID- 16477035 TI - Increased production of reactive oxygen species in hyperglycemic conditions requires dynamic change of mitochondrial morphology. AB - Increased production of mitochondrial reactive oxygen species (ROS) by hyperglycemia is recognized as a major cause of the clinical complications associated with diabetes and obesity [Brownlee, M. (2001) Nature 414, 813-820]. We observed that dynamic changes in mitochondrial morphology are associated with high glucose-induced overproduction of ROS. Mitochondria undergo rapid fragmentation with a concomitant increase in ROS formation after exposure to high glucose concentrations. Neither ROS increase nor mitochondrial fragmentation was observed after incubation of cells with the nonmetabolizable stereoisomer L glucose. However, inhibition of mitochondrial pyruvate uptake that blocked ROS increase did not prevent mitochondrial fragmentation in high glucose conditions. Importantly, we found that mitochondrial fragmentation mediated by the fission process is a necessary component for high glucose-induced respiration increase and ROS overproduction. Extended exposure to high glucose conditions, which may mimic untreated diabetic conditions, provoked a periodic and prolonged increase in ROS production concomitant with mitochondrial morphology change. Inhibition of mitochondrial fission prevented periodic fluctuation of ROS production during high glucose exposure. These results indicate that the dynamic change of mitochondrial morphology in high glucose conditions contributes to ROS overproduction and that mitochondrial fission/fusion machinery can be a previously unrecognized target to control acute and chronic production of ROS in hyperglycemia-associated disorders. PMID- 16477037 TI - Self-fulfilling cavitands: packing alkyl chains into small spaces. AB - The present research was undertaken to determine the conformations of alkyl groups that are forced into small spaces. Unbranched alkyl groups assume fully extended conformations in the free space of solutions, because any bends in the structure create steric clashes between hydrogens along the chain. We synthesized a series of alkyl esters of a carboxylic acid attached to the inner surface of a vase-shaped container structure. The shorter esters (ethyl and propyl) can be accommodated in extended conformations, and even small solvent molecules can share the container's space. Longer (butyl-octyl) esters adopt increasingly coiled conformations that writhe rapidly in the limited space of the cavity. Even longer esters (nonyl and decyl) can be synthesized, but their containers become distorted, and their spectra indicate slowed internal motions of the alkyl groups within the space. In general, alkyl groups are readily contorted when their internal strains are compensated by attraction with the inner surfaces and the proper filling of space. PMID- 16477036 TI - Cooperative transcription activation by Nurr1 and Pitx3 induces embryonic stem cell maturation to the midbrain dopamine neuron phenotype. AB - Midbrain dopamine (DA) neurons play a central role in the regulation of voluntary movement, and their degeneration is associated with Parkinson's disease. Cell replacement therapies, and in particular embryonic stem (ES) cell-derived DA neurons, offer a potential therapeutic venue for Parkinson's disease. We sought to identify genes that can potentiate maturation of ES cell cultures to the midbrain DA neuron phenotype. A number of transcription factors have been implicated in the development of midbrain DA neurons by expression analyses and loss-of-function knockout mouse studies, including Nurr1, Pitx3, Lmx1b, Engrailed 1, and Engrailed-2. However, none of these factors appear sufficient alone to induce the mature midbrain DA neuron phenotype in ES cell cultures in vitro, suggesting a more complex regulatory network. Here we show that Nurr1 and Pitx3 cooperatively promote terminal maturation to the midbrain DA neuron phenotype in murine and human ES cell cultures. PMID- 16477038 TI - Human cytomegalovirus inhibits a DNA damage response by mislocalizing checkpoint proteins. AB - The DNA damage checkpoint pathway responds to DNA damage and induces a cell cycle arrest to allow time for DNA repair. Several viruses are known to activate or modulate this cellular response. Here we show that the ataxia-telangiectasia mutated checkpoint pathway, which responds to double-strand breaks in DNA, is activated in response to human cytomegalovirus DNA replication. However, this activation does not propagate through the pathway; it is blocked at the level of the effector kinase, checkpoint kinase 2 (Chk2). Late after infection, several checkpoint proteins, including ataxia-telangiectasia mutated and Chk2, are mislocalized to a cytoplasmic virus assembly zone, where they are colocalized with virion structural proteins. This colocalization was confirmed by immunoprecipitation of virion proteins with an antibody that recognizes Chk2. Virus replication was resistant to ionizing radiation, which causes double-strand breaks in DNA. We propose that human CMV DNA replication activates the checkpoint response to DNA double-strand breaks, and the virus responds by altering the localization of checkpoint proteins to the cytoplasm and thereby inhibiting the signaling pathway. PMID- 16477039 TI - Automated cell lineage tracing in Caenorhabditis elegans. AB - The invariant cell lineage and cell fate of Caenorhabditis elegans provide a unique opportunity to decode the molecular mechanisms of animal development. To exploit this opportunity, we have developed a system for automated cell lineage tracing during C. elegans embryogenesis, based on 3D, time-lapse imaging and automated image analysis. Using ubiquitously expressed histone-GFP fusion protein to label cells/nuclei and a confocal microscope, the imaging protocol captures embryogenesis at high spatial (31 planes at 1 microm apart) and temporal (every minute) resolution without apparent effects on development. A set of image analysis algorithms then automatically recognizes cells at each time point, tracks cell movements, divisions and deaths over time and assigns cell identities based on the canonical naming scheme. Starting from the four-cell stage (or earlier), our software, named starrynite, can trace the lineage up to the 350 cell stage in 25 min on a desktop computer. The few errors of automated lineaging can then be corrected in a few hours with a graphic interface that allows easy navigation of the images and the reported lineage tree. The system can be used to characterize lineage phenotypes of genes and/or extended to determine gene expression patterns in a living embryo at the single-cell level. We envision that this automation will make it practical to systematically decipher the developmental genes and pathways encoded in the genome of C. elegans. PMID- 16477040 TI - MyD88-dependent and -independent signaling by IL-1 in neurons probed by bifunctional Toll/IL-1 receptor domain/BB-loop mimetics. AB - Interleukin (IL)-1beta is a pluripotent proinflammatory cytokine that signals through the type-I IL-1 receptor (IL-1RI), a member of the Toll-like receptor family. In hypothalamic neurons, binding of IL-1beta to IL-1RI mediates transcription-dependent changes that depend on the recruitment of the cytosolic adaptor protein myeloid differentiation primary-response protein 88 (MyD88) to the IL-1RI/IL-1 receptor accessory protein (IL-1RAcP) complex through homomeric Toll/IL-1 receptor (TIR)-TIR interactions. Through design and synthesis of bifunctional TIR mimetics that disrupt the interaction of MyD88 with the IL 1RI/IL-1RAcP complex, we analyzed the involvement of MyD88 in the signaling of IL 1beta in anterior hypothalamic neurons. We show here that IL-1beta-mediated activation of the protein tyrosine kinase Src depended on a MyD88 interaction with the IL-1RI/IL-1RAcP complex. The activation of the protein kinase Akt/PKB depended on the recruitment of the p85 subunit of PI3K to IL-1RI and independent of MyD88 association with the IL-1RI/IL-1RAcP complex. These bifunctional TIR-TIR mimetics represent a class of low-molecular-weight compounds with both an antiinflammatory and neuroprotective potential. These compounds have the potential to inhibit the MyD88-dependent proinflammatory actions of IL-1beta, while permitting the potential neuronal survival supporting actions mediated by the MyD88-independent activation of the protein kinase Akt. PMID- 16477041 TI - The U(L)41 protein of herpes simplex virus 1 degrades RNA by endonucleolytic cleavage in absence of other cellular or viral proteins. AB - The herpes simplex virus 1 ORF U(L)41 encodes a protein (virion host shutoff or vhs) associated with selective degradation of mRNA early in infection. Some mRNAs, exemplified by GAPDH or beta-actin mRNAs, are degraded rapidly. Others, for example IEX-1 mRNA, are degraded in two stages: whereas the 3' domain disappears rapidly, a large 5' domain fragment of the mRNA lingers for several hours. Still a third, exemplified by tristetraprolin mRNA, is not degraded, allowing its protein product to accumulate in infected cells. Here we report the following: (i) a GST-vhs protein produced in Escherichia coli, solubilized and purified to homogeneity acts as bona fide endoribonuclease when tested on in vitro transcribed IEX-1 probes. A GST-vhs protein in which three key vhs amino acids were replaced with alanines, solubilized and purified by the same protocol, had no enzymatic activity. (ii) The number of fragments generated by cleavage of a truncated IEX-1 RNA by vhs appears to be small; the cleavage sites are centered at or near the AU-rich elements located at the 3' untranslated region of the mRNA. A truncated RNA containing only the IEX-1 coding domain was cleaved numerous times. (iii) In cells infected at high multiplicity and exposed to a large number of particles per cell, the vhs protein accumulated within 3 h after infection, in small uniform cytoplasmic granules raising the possibility that vhs colocalizes with tristerapolin, a protein induced after infection, in structures involved in degradation of RNA. PMID- 16477042 TI - The chromatoid body of male germ cells: similarity with processing bodies and presence of Dicer and microRNA pathway components. AB - The chromatoid body is a perinuclear, cytoplasmic cloud-like structure in male germ cells whose function has remained elusive. Here we show that the chromatoid body is related to the RNA-processing body of somatic cells. Dicer and components of microRNP complexes (including Ago proteins and microRNAs) are highly concentrated in chromatoid bodies. Furthermore, we show that Dicer interacts with a germ cell-specific chromatoid body component, the RNA helicase MVH (mouse VASA homolog). Thus, chromatoid bodies seem to operate as intracellular nerve centers of the microRNA pathway. Our findings underscore the importance of posttranscriptional gene regulation and of the microRNA pathway in the control of postmeiotic male germ cell differentiation. PMID- 16477043 TI - Peptide-matrix-mediated gene transfer of an oxygen-insensitive hypoxia-inducible factor-1alpha variant for local induction of angiogenesis. AB - Hypoxia-inducible factor (HIF) constitutes a target in therapeutic angiogenesis. HIF-1alpha functions as a sensor of hypoxia and induces expression of vascular endothelial growth factor (VEGF), which then induces angiogenesis. To explore the potential of HIF-1alpha gene therapy in stimulating wound healing, we delivered a gene encoding a stabilized form of HIF-1alpha, lacking the oxygen-sensitive degradation domain, namely HIF-1alpha deltaODD, by using a previously characterized peptide-based gene delivery vector in fibrin as a surgical matrix. The peptide vector consisted of multiple domains: (i) A cysteine-flanked lysine hexamer provided DNA interactions that were stable extracellularly but destabilized intracellularly after reduction of the formed disulfide bonds. This DNA-binding domain was fused to either (ii) a fibrin-binding peptide for entrapment within the matrix or (iii) a nuclear localization sequence for efficient nuclear targeting. The HIF-1alpha deltaODD gene was expressed and translocated to the nucleus under normoxic conditions, leading to up-regulation of vascular endothelial growth factor (VEGF)-A165 mRNA and protein levels in vitro. When the peptide-DNA nanoparticles entrapped in fibrin matrices were applied to full-thickness dermal wounds in the mouse (10 microg per wound in 30 microl of fibrin), angiogenesis was increased comparably strongly to that induced by VEGF-A165 protein (1.25 microg per wound in 30 microl of fibrin). However, the maturity of the vessels induced by HIF-1alpha deltaODD was significantly higher than that induced by VEGF-A165 protein, as shown by stabilization of the neovessels with smooth muscle. Nonviral, local administration of this potent angiogenesis-inducing gene by using this peptide vector represents a powerful approach in tissue engineering and therapeutic angiogenesis. PMID- 16477045 TI - Are journals doing enough to prevent fraudulent publication? PMID- 16477047 TI - WTO's new rules allow poorest to import drugs. PMID- 16477046 TI - The vexations of Vioxx. PMID- 16477044 TI - In vitro generated extracellular matrix and fluid shear stress synergistically enhance 3D osteoblastic differentiation. AB - This study instituted a unique approach to bone tissue engineering by combining effects of mechanical stimulation in the form of fluid shear stresses and the presence of bone-like extracellular matrix (ECM) on osteodifferentiation. Rat marrow stromal cells (MSCs) harvested from bone marrow were cultured on titanium (Ti) fiber mesh discs for 12 days in a flow perfusion system to generate constructs containing bone-like ECM. To observe osteodifferentiation and bone like matrix deposition, these decellularized constructs and plain Ti fiber meshes were seeded with MSCs (Ti/ECM and Ti, respectively) and cultured in the presence of fluid shear stresses either with or without the osteogenic culture supplement dexamethasone. The calcium content, alkaline phosphatase activity, and osteopontin secretion were monitored as indicators of MSC differentiation. Ti/ECM constructs demonstrated a 75-fold increase in calcium content compared with their Ti counterparts after 16 days of culture. After 16 days, the presence of dexamethasone enhanced the effects of fluid shear stress and the bone-like ECM by increasing mineralization 50-fold for Ti/ECM constructs; even in the absence of dexamethasone, the Ti/ECM constructs exhibited approximately a 40-fold increase in mineralization compared with their Ti counterparts. Additionally, denatured Ti/ECM* constructs demonstrated a 60-fold decrease in calcium content compared with Ti/ECM constructs after 4 days of culture. These results indicate that the inherent osteoinductive potential of bone-like ECM along with fluid shear stresses synergistically enhance the osteodifferentiation of MSCs with profound implications on bone-tissue-engineering applications. PMID- 16477048 TI - China borrows Canadian know-how for new labs. PMID- 16477049 TI - Challenges for a Conservative minority. PMID- 16477050 TI - Do richer provinces have shorter waiting times to see specialists? PMID- 16477053 TI - Bone infarcts in a woman with systemic lupus erythematosus and antiphospholipid antibody syndrome. PMID- 16477052 TI - Changing epidemiology and emerging risk groups for pertussis. PMID- 16477054 TI - She's dying for help: global HIV. PMID- 16477055 TI - Inflammation at a previous inoculation site: an unusual presentation of Kawasaki disease. PMID- 16477056 TI - Relative value of multiple plasma biomarkers as risk factors for coronary artery disease and death in an angiography cohort. AB - BACKGROUND: Although elevated levels of C-reactive protein (CRP), interleukin (IL)-6, serum amyloid A protein (SAA) and total homocysteine (tHcy) have been associated with the increased likelihood of cardiovascular events, the relative or combined utility of these biomarkers in predicting atherosclerosis and death in an angiography cohort is unknown. METHODS: A cohort of 1117 consecutive patients (797 men and 320 women), referred to 2 Vancouver teaching hospitals for selective coronary angiography, was recruited between 1993 and 1995. Angiography results were obtained for 1019 patients. In 2004 we determined that of 1050 patients who could be traced, 231 had died, 95 of CAD-related causes. We compared the relative utility of baseline measurements of CRP, IL-6, SAA and tHcy as well as of lipids for predicting angiographic CAD and all-cause and CAD-related death. RESULTS: The risk of death increased across quartiles for CRP, IL-6, SAA and tHcy. When comparing the highest and lowest quartiles, the greatest hazard ratios were associated with IL-6 (2.57, 95% confidence interval [CI] 1.62-4.09) and tHcy (2.36, 95% CI 1.53-3.65). A Cox regression model containing all plasma biomarkers and traditional risk factors indicated that age, angiographic CAD and baseline plasma levels of IL-6 and tHcy remained independent predictors of CAD-related death, whereas age, sex, smoking, diabetes and apolipoprotein B levels were independent predictors of angiographic CAD. Kaplan-Meier survival curves indicated a utility in combining measures of CRP, SAA, IL-6 and tHcy for predicting risk of all-cause and CAD-related death. INTERPRETATION: A comparison of elevated levels of CRP, IL-6, SAA and tHcy with traditional CAD risk factors indicated that IL-6 and tHcy were the strongest independent biomarkers for CAD related death. Elevated levels of multiple biomarkers were associated with an increasing rate of all-cause and CAD-related death. PMID- 16477057 TI - Evidence of bias and variation in diagnostic accuracy studies. AB - BACKGROUND: Studies with methodologic shortcomings can overestimate the accuracy of a medical test. We sought to determine and compare the direction and magnitude of the effects of a number of potential sources of bias and variation in studies on estimates of diagnostic accuracy. METHODS: We identified meta-analyses of the diagnostic accuracy of tests through an electronic search of the databases MEDLINE, EMBASE, DARE and MEDION (1999-2002). We included meta-analyses with at least 10 primary studies without preselection based on design features. Pairs of reviewers independently extracted study characteristics and original data from the primary studies. We used a multivariable meta-epidemiologic regression model to investigate the direction and strength of the association between 15 study features on estimates of diagnostic accuracy. RESULTS: We selected 31 meta analyses with 487 primary studies of test evaluations. Only 1 study had no design deficiencies. The quality of reporting was poor in most of the studies. We found significantly higher estimates of diagnostic accuracy in studies with nonconsecutive inclusion of patients (relative diagnostic odds ratio [RDOR] 1.5, 95% confidence interval [CI] 1.0-2.1) and retrospective data collection (RDOR 1.6, 95% CI 1.1-2.2). The estimates were highest in studies that had severe cases and healthy controls (RDOR 4.9, 95% CI 0.6-37.3). Studies that selected patients based on whether they had been referred for the index test, rather than on clinical symptoms, produced significantly lower estimates of diagnostic accuracy (RDOR 0.5, 95% CI 0.3-0.9). The variance between meta-analyses of the effect of design features was large to moderate for type of design (cohort v. case control), the use of composite reference standards and the use of differential verification; the variance was close to zero for the other design features. INTERPRETATION: Shortcomings in study design can affect estimates of diagnostic accuracy, but the magnitude of the effect may vary from one situation to another. Design features and clinical characteristics of patient groups should be carefully considered by researchers when designing new studies and by readers when appraising the results of such studies. Unfortunately, incomplete reporting hampers the evaluation of potential sources of bias in diagnostic accuracy studies. PMID- 16477058 TI - The association between elevated levels of inflammation biomarkers and coronary artery disease and death. PMID- 16477059 TI - Sources of bias in diagnostic accuracy studies and the diagnostic process. PMID- 16477060 TI - Nonpatentable drugs and the cost of our ignorance. PMID- 16477062 TI - Unnecessary exposure? PMID- 16477061 TI - New imaging techniques for diagnosing coronary artery disease. AB - New tomographic cardiovascular imaging tests, such as intravascular ultrasonography (IVUS), coronary computed tomography (CT) angiography and magnetic resonance imaging (MRI), can be used to assess atherosclerotic plaques for the characterization and early staging of coronary artery disease (CAD). Although IVUS images have very high resolution capable of revealing very early preclinical CAD, it is an invasive technique used clinically only in conjunction with a coronary intervention. Multiple-slice coronary CT angiography, which is noninvasive, shows promise as a diagnostic method for CAD. New 64-slice cardiac CT technology has high accuracy for the detection of lesions obstructing more than 50% of the lumen, with sensitivity, specificity, and positive and negative predictive values all better than 90% in patients without known CAD. Cardiac MRI is also improving accuracy in coronary plaque detection and offers a better opportunity for plaque characterization. With further advances in tomographic imaging of coronary atheromas, the goal will be to detect plaques earlier in the development of CAD and to characterize the plaques most likely to generate a clinical event. PMID- 16477063 TI - Training more doctors. PMID- 16477065 TI - Reflections from young physicians. PMID- 16477067 TI - Measuring the presence of chronic diseases. PMID- 16477068 TI - A painful elbow? PMID- 16477069 TI - Reporting communicable diseases. PMID- 16477070 TI - Unnecessary exposure? PMID- 16477073 TI - Is this clinical trial fully registered? PMID- 16477075 TI - Standards of training. PMID- 16477077 TI - An evaluation of class effect. PMID- 16477079 TI - Src homology 2 domain-based high throughput assays for profiling downstream molecules in receptor tyrosine kinase pathways. AB - Src homology 2 (SH2) domains are evolutionary conserved small protein modules that bind specifically to tyrosine-phosphorylated peptides. More than 100 SH2 domains have been identified in proteins encoded by the human genome. The binding specificity of these domains plays a critical role in signaling within the cell, mediating the relocalization and interaction of proteins in response to changes in tyrosine phosphorylation states. Here we developed an SH2 domain profiling method based on a multiplexed fluorescent microsphere assay in which various SH2 domains are used to probe the global state of tyrosine phosphorylation within a cell and to screen synthetic peptides that specifically bind to each SH2 domain. The multiplexed, fluorescent microsphere-based assay is a recently developed technology that can potentially detect a wide variety of interactions between biological molecules. We constructed 25-plex SH2 domain-GST fusion protein conjugated fluorescent microsphere sets to investigate phosphorylation-mediated cell signaling through the specific binding of SH2 domains to activated target proteins. The response of HeLa, COS-1, A431, and 293 cells and four breast cancer cell lines to epidermal growth factor and insulin were quantitatively profiled using this novel microsphere-based, multiplexed, high throughput assay system. PMID- 16477080 TI - Thematic review series: lipid posttranslational modifications. Structural biology of protein farnesyltransferase and geranylgeranyltransferase type I. AB - More than 100 proteins necessary for eukaryotic cell growth, differentiation, and morphology require posttranslational modification by the covalent attachment of an isoprenoid lipid (prenylation). Prenylated proteins include members of the Ras, Rab, and Rho families, lamins, CENPE and CENPF, and the gamma subunit of many small heterotrimeric G proteins. This modification is catalyzed by the protein prenyltransferases: protein farnesyltransferase (FTase), protein geranylgeranyltransferase type I (GGTase-I), and GGTase-II (or RabGGTase). In this review, we examine the structural biology of FTase and GGTase-I (the CaaX prenyltransferases) to establish a framework for understanding the molecular basis of substrate specificity and mechanism. These enzymes have been identified in a number of species, including mammals, fungi, plants, and protists. Prenyltransferase structures include complexes that represent the major steps along the reaction path, as well as a number of complexes with clinically relevant inhibitors. Such complexes may assist in the design of inhibitors that could lead to treatments for cancer, viral infection, and a number of deadly parasitic diseases. PMID- 16477081 TI - Differentiation-associated expression of ceramidase isoforms in cultured keratinocytes and epidermis. AB - Ceramides (Cers) accumulate within the interstices of the outermost epidermal layers, or stratum corneum (SC), where they represent critical components of the epidermal permeability barrier. Although the SC contains substantial sphingol, indicative of ceramidase (CDase) activity, which CDase isoforms are expressed in epidermis remains unresolved. We hypothesized here that CDase isoforms are expressed within specific epidermal compartments in relation to functions that localize to these layers. Keratinocytes/epidermis express all five known CDase isoforms, of which acidic and alkaline CDase activities increase significantly with differentiation, persisting into the SC. Conversely, neutral and phytoalkaline CDase activities predominate in proliferating keratinocytes. These differentiation-associated changes in isoform activity/protein are attributed to corresponding, differentiation-associated changes in mRNA levels (by quantitative RT-PCR). Although four of the five known CDase isoforms are widely expressed in cutaneous and extracutaneous tissues, alkaline CDase-1 occurs almost exclusively in epidermis. These results demonstrate large, differentiation-associated, and tissue-specific variations in the expression and activities of all five CDase isoforms. Because alkaline CDase-1 and acidic CDase are selectively upregulated in the differentiated epidermal compartment, they could regulate functions that localize to the distal epidermis, such as permeability barrier homeostasis and antimicrobial defense. PMID- 16477083 TI - Neuropathology in the S305S tau gene mutation. PMID- 16477084 TI - Reply to 'Mitochondrial changes in skeletal muscle in amyotrophic lateral sclerosis and other neurogenic atrophies--a comment'. PMID- 16477087 TI - Making progress on the natural history of multiple sclerosis. PMID- 16477088 TI - Medial reorganization of motor function demonstrated by functional MRI and diffusion tensor tractography. AB - PURPOSE: Peri-lesional reorganization is a motor recovery mechanism after brain injury. The object of this study was to demonstrate the peri-lesional reorganization, using functional MRI (fMRI) and diffusion tensor tractography (DTT). METHODS: Six control subjects and a 53 year-old woman with left primary sensori-motor cortex (SM1) and underlying deep white matter injury were evaluated. The patient presented with complete paralysis of the right hand after the resection of a meningioma on the left SM1. The motor function of the affected hand had recovered slowly the extent of her being able to overcome some resistance for 6 months. At 2 years after the operation, fMRI was performed at 1.5 T in parallel with timed finger flexion-extension movements in all subjects and DTT was performed only in the patient. RESULTS: The contralateral SM1 centered on the precentral knob was activated during hand movements of unaffected (left) side or control subjects. However, the medial area of the injured SM1 was activated during affected (right) hand movements. DTT of the affected (left) hemisphere originated from the medial area of the injured SM1 and descended along the medial side of injured deep white matter. CONCLUSIONS: It seems that the motor function of the affected hand was reorganized into the medial area of the injured SM1 and deep white matter in this patient. PMID- 16477089 TI - Amphetamine fails to facilitate motor performance and to enhance motor recovery among stroke patients with mild arm paresis: interim analysis and termination of a double blind, randomised, placebo-controlled trial. AB - PURPOSE: To assess the effects of d-amphetamine on motor facilitation and recovery in stroke patients with mild arm paresis receiving the Arm Ability training. METHODS: Thirty-one stroke patients with mild arm paresis were randomly assigned to either (a.) receiving placebo or (b.) d-amphetamine twice a week 2 hours before Arm Ability training sessions for three weeks. MAIN OUTCOME MEASURE: time needed to perform TEMPA tasks, a measure of upper extremity performance reflecting everyday life tasks. Secondary measures: aimed movements, tapping, and a 10 m walkway as well as motor performance during training sessions. RESULTS: The interim efficacy analysis of 26 stroke patients who completed the study intervention showed overall arm motor recovery both from pre to post test after 3 weeks of training and from pre test to follow-up 1 year later. No superior effect of d-amphetamine over placebo could be substantiated for either motor facilitation during training or motor recovery (post training or long-term). D amphetamine exerted mild effects on blood pressure. Serious adverse events were not observed. CONCLUSIONS: d-Amphetamine failed to facilitate motor performance during training sessions, to promote skill acquisition with training tasks, and most importantly to enhance motor recovery among patients with mild arm paresis after stroke. PMID- 16477090 TI - Assessment of auditory temporal-order thresholds - a comparison of different measurement procedures and the influences of age and gender. AB - PURPOSE: The relationship between auditory temporal-order perception and phoneme discrimination has been discussed for several years, based on findings, showing that patients with cerebral damage in the left hemisphere and aphasia, as well as children with specific language impairments, show deficits in temporal-processing and phoneme discrimination. Over the last years several temporal-order measurement procedures and training batteries have been developed. However, there exists no standard diagnostic tool for adults that could be applied to patients with aphasia. Therefore, our study aimed at identifying a feasible, reliable and efficient measurement procedure to test for auditory-temporal processing in healthy young and elderly adults, which in a further step can be applied to patients with aphasia. METHODS: The tasks varied according to adaptive procedures (staircase vs. maximum-likelihood), stimuli (tones vs. clicks) and stimulation modes (binaural- vs. alternating monaural) respectively. A phoneme-discrimination task was also employed to assess the relationship between temporal and language processing. RESULTS: The results show that auditory temporal-order thresholds are stimulus dependent, age related, and influenced by gender. Furthermore, the cited relationship between temporal-order threshold and phoneme discrimination can only be confirmed for measurements with pairs of tones. CONCLUSION: Our results indicate, that different norms have to be established for different gender and age groups. Furthermore, temporal-order measurements with tones seem to be more suitable for clinical intervention studies than measurements with clicks, as they show higher re-test reliabilities, and only for measurements with tones an association with phoneme-discrimination abilities was found. PMID- 16477091 TI - Training the "less-affected" forelimb after unilateral cortical infarcts interferes with functional recovery of the impaired forelimb in rats. AB - PURPOSE: Unilateral lesions of the sensorimotor cortex (SMC) in adult rats cause major behavioral changes in the ipsilesional, "less-affected" forelimb. An increase in function and reliance on this forelimb can aid compensation for contralesional impairments, but may also promote disuse and reduced functionality of the impaired forelimb. We hypothesized that training focused on the ipsilesional forelimb following a unilateral SMC lesion would reduce the efficacy of later motor rehabilitative training of the impaired forelimb. METHODS: Rats with ischemic SMC lesions were trained on a skilled reaching task with the ipsilesional forelimb (PriorT) or received control procedures (Cont) for 10 days. Both groups were then trained with the impaired forelimb on the same reaching task for 10 days. RESULTS: In comparison with Cont, PriorT rats had little improvement on the reaching task with the impaired forelimb and had a more enduring disuse of the impaired forelimb for postural support behaviors. Lesion sizes were similar between groups. CONCLUSIONS: Behavioral experience with the less-affected forelimb early after unilateral SMC lesions has the potential to increase disuse and dysfunction of the impaired forelimb, consistent with a training-induced exacerbation of learned non-use. These findings are suggestive of competitive processes in experience-dependent neural restructuring after brain damage. PMID- 16477092 TI - Recovery of visual evoked potentials after regeneration of cut retinal ganglion cell axons within the ascending visual pathway in adult rats. AB - PURPOSE: Following optic nerve damage, retinal ganglion cells (RGCs) fail to regenerate their axons and soon undergo apoptosis. However, many RGCs survive axotomy and regenerate lengthy axons after a lens injury (LI). If the cut optic nerve is re-sutured, RGC axons grow into the distal part of the optic nerve and reach their natural targets within the thalamus and midbrain. In this study, we check time-dependence and extent of restoration of flash visual evoked potentials (FVEPs) to examine the functional relevance of the regenerated retinogeniculate pathway. METHODS: The optic nerve in adult rats was cut and re-sutured. The lens was injured transsclerally using a pointed glass capillary. FVEPs were measured starting at the time point of surgery, and then repeatedly up to an age of several months. RESULTS: Detectable FVEPs appeared approximately ten weeks after the surgery, and their amplitudes increased during the next months to reach eventually 15-40% of their values before surgery. CONCLUSIONS: Partial restoration of FVEPs indicates that some regenerating RGC axons have "bridged" the distance between the eye and the central targets forming a functional re connection of the corresponding RGC with thalamic target neurones to elicit recordable activation of the visual cortex. PMID- 16477093 TI - Lentiviral vector-mediated transduction of neural progenitor cells before implantation into injured spinal cord and brain to detect their migration, deliver neurotrophic factors and repair tissue. AB - PURPOSE: Stem cells represent an attractive source for cell replacement therapy in neurological disorders due to their self-renewal and multi-potency. Genetic manipulation of these cells may allow controlled release of therapeutic proteins, suppress immune rejection, or produce essential neurotransmitters. Furthermore, when the expression cassette is incorporated into the host genome ex vivo, this technique also may be used as a method to trace cells following implantation into tissues of interest. METHODS: We explored the possibility of transducing pluripotent fetal rat cortical neural progenitor cells (NPCs) using lentiviral vectors encoding the green fluorescent protein (GFP) or neurotrophic factors (BDNF, CNTF, D15A, GDNF, MNT and NT-3) prior to implanting these cells into the contused spinal cord or injured brain. RESULTS: In vitro staining of these cells for neural markers (such as nestin, GFAP, Tuj-1 and RIP) after transduction did not reveal any significant difference from non-transduced cells. When they were transduced with a vector encoding CNTF or MNT, however, cells started expressing GFAP in vitro. Following delayed (1 week) implantation into the lesion site of the moderately contused rat spinal cord or the injured brain, transduced cells survived up to 12 weeks post-implantation (the longest time point examined) and most of the NPCs turned into an astrocytic phenotype in the spinal cord, but not in the brain. Nestin and GFP positive cells were detected in the brain, but not in the spinal cord lesion. GFP positive cells in the spinal cord migrated rostrally and caudally from the lesion/implantation site towards uninjured tissue. CONCLUSIONS: Novel findings in this study are the longterm expression of a foreign gene in NPCs using lentiviral vectors; this enabled tracking of the cells following implantation. This expression also allowed the observation that NPCs developed differently in the injured spinal cord and brain. Moreover, NPCs could be transduced to overexpress neurotrophic factors. In sum, NPC survival and the long-term transgene expression that allows easy tracking of migrating cells make NPCs promising candidates for implantation into the injured spinal cord or brain and a potentially powerful tool to enhance regeneration when transduced ex vivo to produce therapeutic molecules. PMID- 16477094 TI - Cortical reorganization associated lower extremity motor recovery as evidenced by functional MRI and diffusion tensor tractography in a stroke patient. AB - PURPOSE: Recovery mechanisms supporting upper extremity motor recovery following stroke are well established, but cortical mechanism associated with lower extremity motor recovery is unknown. The aim of this study was to assess cortical reorganization associated with lower extremity motor recovery in a hemiparetic patient. METHODS: Six control subjects and a 17 year-old woman with left intracerebral hemorrhage due to an arterio-venous malformation rupture were evaluated. The motor function of the paretic (left) hip and knee had recovered slowly to the extent of her being able to overcome gravity for 10 months after the onset of stroke. However, her paretic upper extremity showed no significant motor recovery. Blood oxygenation level dependent (BOLD) functional MRI at 1.5 Tesla was used to determine the acutual location of cortical activation in the predefined regions of interest. Concurrently, Diffusion Tensor Imaging (DTI) in combination with a novel 3D-fiber reconstruction algorithm was utilized to investigate the pattern of the corticospinal pathway connectivity between the areas of the motor stream. All subjects' body parts were secured in the scanner and performed a sequential knee flexion-extension with a predetermined angle of 0 60 degrees at 0.5 Hz. RESULTS: Controls showed anticipated activation in the contralateral sensorimotor cortex (SM1) and the descending corticospinal fibers stemming from motor cortex. In contrast to control normal subjects, the stroke patient showed fMRI activation only in the unaffected (right) primary SM1 during either paretic or nonparetic knee movements. DTT fiber tracing data showed that the corticospinal tract fibers were found only in the unaffected hemisphere but not in the affected hemisphere. CONCLUSIONS: Our results indicate that an ipsilateral motor pathway from the unaffected (right) motor cortex to the paretic (right) leg was present in this patient. This study raises the potential that the contralesional (ipsilateral) SM1 is involved in cortical reorganization associated lower extremity motor recovery in stroke. This study is the first neuroimaging evidence that the combined fMRI and DTI fiber tracing can significantly expand the explanatory power of probing cortical reorganization underlying motor recovery mechanism in stroke. PMID- 16477095 TI - The ability of physiological stimuli to generate the sympathetic skin response in human chronic spinal cord injury. AB - PURPOSE: Sympathetic sudomotor function in chronic spinal cord injury (SCI) has been evaluated to determine if recording the sympathetic skin response (SSR) provides evidence of integrity of the spinal component of the sympathetic pathways. METHODS: Thirty subjects with chronic SCI and 15 healthy normal subjects were studied. The SSR was elicited using two physiological (auditory and inspiratory gasp) stimuli. In addition, electrical (median and peroneal nerve) stimulation was also performed. Recordings were made from palmar and plantar sites. RESULTS: Palmar and plantar SSRs could be readily elicited in all control subjects by all stimuli. In the majority of SCI subjects, the presence or absence of the SSR was related to the American Spinal Injury Association (ASIA) impairment scale, which incorporates only motor and sensory assessments. The exceptions indicated preserved (or damaged) sympathetic spinal cord pathways. CONCLUSIONS: We conclude that the SSR, using either physiological or electrical stimuli, may be a reliable, non-invasive method of determining integrity of sympathetic cholinergic pathways in SCI, with potential for monitoring the effects of intervention and spinal repair. PMID- 16477096 TI - Erythropoietin enhances neurological recovery after experimental spinal cord injury. AB - PURPOSE: Erythropoietin is a hematopoietic growth factor. It is also involved in neurodevelopment and neuroprotection. The purpose of this study was to evaluate the effectiveness of erythropoietin in enhancing the neurological recovery following experimental spinal cord injury. METHODS: Rats were randomly divided into 4 groups. Group 1 received only laminectomy. Group 2, 3, and 4 have undergone a spinal contusion injury of 50 gr/cm. Group 2 received no medication. Group 3 received 30 mg/kg methylprednisolone. Group 4 received 5000 IU/kg recombinant human erythropoietin. Following injury, neurological recovery was evaluated for 14 days, using a swimming test. RESULTS: At day 1, there was no difference between mean motor scores of group 2, 3, and 4. Following day 1, rats in group 4 exhibited a marked improvement in motor score, and this was maintained throughout the study. When compared to group 2, mean motor score of group 4 was significantly higher at day 4, 7, 10, and 14. When compared to group 3, mean motor score of group 4 was significantly higher at day 7, 10, and 14. CONCLUSION: These findings suggest that erythropoietin enhances neurological recovery following spinal cord injury and it seems to be more effective than methylprednisolone at the given doses. PMID- 16477097 TI - Alterations in conduction characteristics of crushed peripheral nerves. AB - PURPOSE: Nerve crush injury results in the incapability to maintain conduction of an impulse. Disruption in the myelin sheath causes very important changes in the activities of ion channels. Therefore, crushed and intact sciatic nerves were researched with both histological and electrophysiological methods in this study. METHODS: Electron and light microscopy for histological examinations, a sucrose gap recording techniques for electrophysiological examinations were used. 4 aminopyridine (4-AP) and Tetraethylammonium (TEA) were used to functional separation of the fast and slow K;+ channels. RESULTS: The number of damaged myelinated nerve fiber was counted as 750 +/- 3.5. Lamellar separation and disruption in myelin sheath was frequently observed in these fibers. Conduction velocity of crushed nerves (19 +/- 2 m/s) was half of the intact nerves. The relationship between equally spaced interstimulus interval and their responses demonstrated that 4-AP and 4-AP plus TEA have more pronounced effects on crushed nerves than on intact nerves. After 4-AP, TEA application caused an efficiently depolarization in the membrane potential. CONCLUSIONS: The effects of 4-AP and 4 AP plus TEA suggest the involvement of slow and fast K;+ channels and slow Na;+ currents in membrane potential and action potential repolarization. Minimal myelin damage may significantly influence the subsequent impulse generation and the patterning of action potential activity. PMID- 16477098 TI - RAGE: a journey from the complications of diabetes to disorders of the nervous system - striking a fine balance between injury and repair. AB - The Receptor for Advanced Glycation End Products (RAGE) is a multiligand member of the immunoglobulin superfamily. RAGE interacts with AGEs, the products of nonenzymatic glycation/oxidation of proteins and lipids that accumulate in diverse settings, such as diabetes, inflammation, renal failure, pro-oxidant states and natural aging. In addition, RAGE is also a receptor for amyloid-beta peptide and beta-sheet fibril species. Recent studies underscore the premise that RAGE interacts with pro-inflammatory molecules, including S100/calgranulins and amphoterin, the latter also known as high mobility group box 1 (HMGB1). In chronic neurodegenerative disorders as well as in nerve tissue upon acute injury, evidence points to upregulation of both RAGE and these ligand families. In this review, we will discuss the implications of transient/self-limited upregulation of RAGE and its ligands, vs sustained/chronic upregulation of this axis in neurodegeneration vs repair in both the central and peripheral nervous systems. Experimental evidence supports the premise that RAGE bears both homeostatic and injurious properties in the nervous system, thereby highlighting "yin/yang" features of this receptor and its ligand families. PMID- 16477101 TI - Factors influencing the slip index measurements with the Horizontal Pull Slipmeter. AB - Several factors that might influence the outcomes of the slip index measurements with the Horizontal Pull Slipmeter were examined. These factors were either those not clearly specified in the operating guidelines of the device or those clearly specified but with a high variability in operator compliance. The results indicated that the shoe pad thickness, sanding, alignment between the string and the force gauge, and temperature were the factors that led to significant differences at the levels evaluated. The differences due to string stiffness, string length, duration from cleaning to friction measurement, dwell time, battery level and relative humidity did not reach a statistically significant level. Extra guidelines are needed for those factors that could result in significant variations in order to lead to more consistent results. PMID- 16477099 TI - Emerging strategies to promote improved functional outcome after peripheral nerve injury. AB - PURPOSE: To present a general review of experimental strategies used to improve functional outcome after peripheral nerve injury. In order to understand the mechanisms behind the strategies, the process of nerve healing after injury is described briefly and each strategy is described in its context. Since the functional outcome is not solely determined by nerve regeneration but by a number of different factors, we have also chosen to cover many other important topics. METHODS: Literature review. RESULTS: Review article. CONCLUSIONS: Functional outcome after peripheral nerve injury is often poor and sometimes associated with neuropathic pain. Therapeutic intervention can be carried out at different levels and we attempt to place the different strategies and target molecules, in the context of the nerve healing process. The most obvious interventions are perhaps to minimize cell death and enhance regeneration across the lesion gap. Others, which are more difficult, may be to limit neuropathic pain, improve target finding and cortical reorganization, counteract effects of prolonged denervation/axotomy, and reduce electrical conduction block at the scar formation. Although clinical outcome is often poor at present, recent preclinical research provides several promising approaches and new target molecules for therapeutic intervention, such as neurotrophic factor (GDNF and ARTN) treatment of neuropathic pain, manipulation of the small Rho GTPases (Rac, Rho, Cdc42, Tc10), lipid raft manipulation, gene silencing with DNA enzyme and siRNA. In addition, recent research involving high throughput screening of gene expression (microarray) after nerve injury encourages the discoveries of new possible target molecules. PMID- 16477102 TI - A controlled case study of supervisor training to optimize response to injury in the food processing industry. AB - The role of supervisors to aid injured workers, access health care, and provide reasonable accommodation may prevent prolonged disability among workers reporting musculoskeletal pain. Although supervisor training has been a common element of broad-based ergonomic interventions to prevent injuries, the impact of supervisor training alone to improve injury response has not been studied. In a controlled design, 11 supervisors in an intervention group and 12 supervisors in a delayed intervention control group from the same plant were provided a 4-hour training workshop. The workshop emphasized communication skills and ergonomic accommodation for workers reporting injuries or health concerns. Workers' compensation claims data in the 7 months before and after the workshop showed a 47% reduction in new claims and an 18% reduction in active lost-time claims versus 27% and 7%, respectively, in the control group. Improving the response of frontline supervisors to employees' work-related health and safety concerns may produce sustainable reductions in injury claims and disability costs. PMID- 16477103 TI - The effect of shiftwork related fatigue on the family life of train operators: implications for safety and health professionals. AB - Drawing upon an original research study about the effects of fatigue on train operators, the present article focuses upon family issues as having the most significant impact on the participants. Family support, for example, represents an important mechanism for managing and coping with fatigue. Family support comprises understanding of the physiological and emotional issues surrounding shiftwork and erratic work schedules. This article explores the impact of fatigue upon a variety of family and relational issues. This inquiry describes the impact of family on the broader employee view of managing fatigue that considers such comprehensive issues as perception of the work environment, emotional stability, personal control concerns and other positive attributes. These issues ultimately impact the health, productivity and performance of employees. Final discussion includes implications for workplace application of the research findings. PMID- 16477104 TI - Reliability of normalisation methods for EMG analysis of neck muscles. AB - Acceptable reliability of normalisation contractions in electromyography (EMG) is paramount for testing conducted over a number of days or if normal laboratory strength testing equipment is unavailable. This study examined the reliability of maximal voluntary isometric contractions (MVIC) and sub-maximal (60%) isometric contractions for use in neck muscle EMG studies. Surface EMG was recorded bilaterally from eight sites around the neck at C4/5 level from five healthy male subjects. Subjects performed MVIC and sub-maximal normalisation contractions using an isokinetic dynamometer (ID) and a portable cable dynamometer with attached strain gauge (PCD) in addition to a MVIC against a manual resistance (MR). Subjects were tested in flexion, extension, left and right lateral bending and were retested by the same tester within a two-week period. Intra class correlation co-efficients (ICC) were calculated for each testing method and contraction direction and a mean ICC was calculated across all contraction directions. All normalisation methods produced excellent within-day reliability (mean ICC >0.80) but only the MVICs using the ID and PCD had acceptable reliability when assessed between-days. This study confirmed the validity of using MVICs elicited using the ID and PCD as reliable reference contractions for the normalisation of neck EMG. PMID- 16477105 TI - Reliability of the instrument DOA: dialogue about ability related to work. AB - The aim of this study was to examine the reliability of the new instrument "DOA- a dialogue about ability related to work" [6]. DOA was developed using the Model of Human of Occupation as a theoretical base [12,13]. The instrument is divided into two sections; client self-assessment and professional assessment focusing on the individual's ability to perform work-related activities. Assessments are followed by a dialogue to distinguish goals for the return-to-work process based on the client's own preferences [6,14]. 34 outpatients' clients and 14 professionals from four clinics within psychiatric work rehabilitation in Sweden participated in the study. The Spearman Rank Correlation test generally showed acceptable significant correlations in test-retest (n=34). A greater part of items correlated "moderate to good" (r=0.51-0.75). In the raters' test (n=14) Percentage of agreement (PA) was good ranging between 93.1%-96.5%, which is above the acceptable criteria of 80%. There were no significance differences between the raters. The study has indicated that items in DOA work with generally good reliability to measure ability related to work. The instrument was demonstrated to be a reliable therapeutic tool in goal setting for return-to-work in a position outside of the clients' preferences. PMID- 16477106 TI - Postural control in nurses with and without low back pain. AB - BACKGROUND: Low back pain (LBP) is common in nurses, but it is unclear if the pain is associated with measurements of postural control. Objective measurements of function might be useful in the evaluation of patients with LBP in order to evaluate and predict disability, and in the study of the pathophysiology of chronic LBP. METHODS: In a cross-sectional study, we measured the number of postural adjustments, and degree of posterior-lateral sway in 81 nurses, using a computerized postural sway four-platter measurement system. RESULTS: There were 41 (56.6%) nurses who complained of LBP at the time of testing, and another 12 (14.8%) with a past history of LBP. Nurses with LBP consistently used more postural adjustments to keep their balance (p<0.003), and in some positions postural adjustments were positively associated with the degree of past and present pain. LBP was not significantly associated with the degree of lateral sway. CONCLUSIONS: Nurses either with present or a past history of LBP use an increased number of postural adjustments to maintain balance. Studies are warranted to determine if postural testing can predict the development of LBP or aid in determining appropriate preventive measures. PMID- 16477107 TI - Return to work still possible after several years as a disability pensioner due to musculoskeletal disorders: a population-based study after new legislation in Sweden permitting "resting disability pension". AB - Different strategies have been used to stimulate a return to work (RTW) among individuals suffering from long-term ailments. In Sweden a new law on "resting disability pension" permits disability pensioners to go back to work without jeopardising their benefits. In this study different variables related to RTW during 2000 by means of this legislation were identified among disability pensioners with musculoskeletal disorders. Individuals in the study group, when compared to a control group, had more often been disability pensioners for several years, had additional education, estimated their previous job to have been physically strenuous to a lesser degree, were more satisfied with the treatment at the social insurance office and had a more positive self-image. This study shows that it may be meaningful to continue/resume rehabilitation efforts and to try to motivate an individual suffering from musculoskeletal disorders to return to work even after several years as a disability pensioner. PMID- 16477108 TI - Conditions that influence the elimination of postural constraints after office employees working with VDU have received ergonomics training. AB - OBJECTIVE: The goal of this article is to better understand how preventive measures are undertaken after training. It examines how certain variables, such as musculoskeletal pain, participant age and workstation and work content characteristics influence the reduction of postural constraints after office employees working with a computer have received ergonomics training. METHODS: A pre-test/post-test design was used. The 207 female office workers were given 6 hours of ergonomics training. The variables were determined using a self administered questionnaire and an observation grid filled out 2 weeks before and 6 months after the training session. The FAC and HAC were used in the data processing. RESULTS: The presence or absence of musculoskeletal pain had no statistically significant influence on whether or not postural constraints were eliminated. The age of the participants and the possibility of adjusting the workstation characteristics and work content produced differentiated results with regard to postural constraint reduction. We concluded that trained people succeed in taking relevant and effective measures to reduce the postural constraints found in VDUs. However other measures than work station adjustments lead to this prevention and such training must be strongly supported by the various hierarchical levels of an enterprise or an institution. PMID- 16477109 TI - Health-related determinants of perceived quality of life: a comparison between first-year university students and their working peers. AB - This study compares aspects of the health-related behaviors and working life of first-year university students with those of their working counterparts and assesses the roles of these aspects as predictors of each groups' perceived quality of life (QoL). Subjects were taken from two cross-sectional data sets (a student survey and a population survey) from the Swedish central-western region of Ostergotland. Male and female respondents aged 20-35 years were extracted and comparison were made considering in turn socio-demographic characteristics, health-related behaviors, exposure to abusive events at work and perceived QoL. University students smoked and used oral snuff in smaller proportions, they were not as frequent drinkers as their working counterparts, but they tended to drink more when they did drink. Threats and violence were less prevalent among students, but sexual harassment, were almost as common. The strongest predictors of perceived QoL for both groups are expected and former perceived QoL, followed by current self-rated health. PMID- 16477110 TI - Factors relevant to a return to work: a multivariate approach. AB - This paper contributes to the understanding of the factors that are significant for returning to work, and identifies factors that might be used early on in a period of sick leave to discern whether people are likely to work again. In the design, the multivariate Partial Least Squares (PLS) of Latent Structures method was used to analyse information from a questionnaire containing socio-demographic items, and information on symptoms, consequences on daily life, expectations and psychosocial factors. Data about the incidence of sickness over a four-year period was included. 121 adults aged 18-64 years on sick leave participated, irrespective of their diagnoses. A reliable prediction of a return to work required the combination of many factors: individual psychosocial instruments are not useful when considered in isolation. The strongest predictive factors for a return to work concern the individuals' expectations, the number of days of sick leave taken in the past, somatic disorders, and a high level of life satisfaction and sense of coherence. Many factors influence the outcome for people on sick leave: PLS analysis demonstrated that a multivariate approach using this method could predict the long-term outcome early on in a period of sick leave. PMID- 16477111 TI - How can cooperation between rehabilitation professionals in rehabilitation planning be improved? A qualitative study from the employer's perspective. AB - Research shows that there are often problems with cooperation between rehabilitation professionals within vocational rehabilitation. The aim of this study was to describe employers' experiences of how cooperation between different rehabilitation professionals can be improved in a vocational rehabilitation planning process. Ten employers who had sent their employees to vocational rehabilitation at a rehabilitation centre in the north of Sweden during 2000 and 2001 participated in the study. Qualitative interviews were performed and analysed by thematic content analysis. The employers' cooperation with clients could be improved by a focus on clients' needs and participation in the rehabilitation process. The employers cooperation with Social Insurance companies could be improved by, 1) early prevention and intervention, 2) knowledge of each other's roles, responsibilities and opportunities and 3) priority making. The employers' cooperation with rehabilitation professionals could be improved by, 1) early identification of rehabilitation needs and goals and early rehabilitation and, 2) increased focus on own responsibilities from each part in the process. PMID- 16477112 TI - Gender-age environmental associates of middle school students' low back pain. AB - PURPOSE: The purpose of this study was to describe adolescents' low back pain, investigate its prevalence and explore its associations with backpack load, time spent carrying loads, time sitting, and time playing sports, to determine whether relationships exist among these variables. METHOD: One hundred urban middle school students in Manchester, New Hampshire completed a questionnaire (55% response rate). Participants were between 13-14 years old. Weights of students, loaded backpacks, backpack contents, and students' heights were measured separately. RESULTS: Eighty-nine percent of the participants wore the two-straps backpack. Over eighty percent of them preferred to carry the load over two shoulders. The average load weighed 4.9 kg (approximating 9.6% of the participants' body weight). There was a significant association found between backpack carrying time and adolescents' low back pain. CONCLUSION: Daily backpack carrying is a frequent cause of musculoskeletal discomfort for adolescents. The association between backpack carrying time and low back pain may provide the impetus for parents, teachers, and school administrators to decrease the prolonged carrying of backpacks. Further investigations should be conducted to evaluate the intervention effectiveness after implementation of controls. PMID- 16477113 TI - A research pilot project to test the efficacy of current pedorthic practices in the Canadian Forces. AB - Impact loading is a major factor in the high prevalence of musculoskeletal injuries among military personnel during operational tasks. One of the therapeutic purposes of a pedorthic approach is to attenuate impact load through footwear and supportive device cushioning. The following research pilot project attempts to investigate this relationship. Vertical acceleration was collected at the fourth lumbar level in 30 members of the Canadian Forces (CF) during two ladder descents on a CF warship. Body weight, time of descent, and a description of footwear and any additional supportive appliances were also collected. None of the evaluated variables were significantly different between members wearing standard issue footwear and medically prescribed footwear and supportive devices. Although the results of this research pilot must be viewed with caution, the preliminary findings of this pilot tends to suggest that the current pedorthic practices may not effectively reduce peak impact forces at the fourth lumbar level of the spine among CF personnel during a selected naval activity. PMID- 16477114 TI - A study of the effects of isokinetic pre-employment physical capability screening in the reduction of musculoskeletal disorders in a labor intensive work environment. AB - OBJECTIVES: This study investigated the effects of pre-employment physical ability screening using isokinetic dynamometry in injury development, specific to musculoskeletal disorders (MSDs) of the knees, shoulders and back among workers in physically demanding jobs. METHODS: New hires (n=503) from a large US employer's 105 industrial yards were screened to match the physical demands of their prospective jobs and tracked for up to 33 months. Results were compared to a control group of 1423 workers. RESULTS: There were significant reductions in the frequency and severity of musculoskeletal disorder injuries in the screened employee population. Non-screened applicants were 2.38 times more likely to experience a MSD-related overexertion injury specific to the knees, shoulders and back than screened hires (OR=2.3759; p=0.0001), and incurred 4.33 times higher cost of claims (p=0.0003). Covariates of age, pay type, race and job classification were markedly different between screened and unscreened hires. Among the screened cohort, only the more physically demanding job classifications were significant with field material handlers 7.1 times more likely to experience a non-MSD than less physically demanding workers (OR=7.1036; p=0.0063). CONCLUSIONS: Objective isokinetic pre-employment screening may significantly reduce injuries in physically demanding jobs. Employees having been effectively matched to the physical demands of their jobs may be at significantly lesser risk of injury and disability from both musculoskeletal and non-musculoskeletal disorders. PMID- 16477115 TI - Item validity of the physical demands from the Dictionary of Occupational Titles for FCE of clients with chronic back pain. PMID- 16477117 TI - Chemical surface modification of high-strength porous Ti compacts by spark plasma sintering. AB - The biological properties of a titanium (Ti) implant depend on its surface oxide film. The aims of the present study were to increase the specific surface oxide area on Ti using a porous structure and to study the relationship between the amount of apatite coating in simulated body fluid (SBF) and the actual surface area on titanium powders. Ti powders of 110 microm average diameter were sintered by spark plasma sintering. The sintered compacts had a porosity of 28%, a compressive elastic modulus of 7.9 GPa and an ultimate strength of 112 MPa. The compressive strength of the compacts was increased to 588 MPa by subsequent annealing in a vacuum furnace at 1000 degrees C for 24 h. The sintered compacts were treated with aqueous NaOH solution and subsequently heated at 600 degrees C. The pretreated compacts showed apatite crystal precipitation in SBF. The amounts of precipitates through the compacts were compared with those of the Ti plate substrates subjected to the same chemical pretreatment. It was confirmed that the amounts of precipitates through the compacts were more than one hundred times higher than those on the Ti plates. It was concluded that the metal porous compacts developed may be used as functional materials for immobilizing functional proteins and/or drugs, because the precipitated apatite can adsorbed these substances. PMID- 16477118 TI - An in vitro investigation of the effect of the addition of untreated and surface treated silica on the transverse and impact strength of poly(methyl methacrylate) acrylic resin. AB - Silica is a commonly used filler in dental materials and as a reinforcing agent in industry. The aim of this study was to further investigate the effect of the addition of untreated and a novel surface treated silica on the transverse bend and impact strength of acrylic resin denture base material. It was hypothesized that the silica/resin composite materials would have an improved flexural and impact strength than the conventional heat-cured acrylic resin. Three types of untreated and two of treated silica powder were used in this study. The range of percentages used were 1%, 0.5%, 0.2%, 0.1%. The treated particles were coated with hexamethyldisilazane or dimethyldichloridesilazane. Conventional heat cured acrylic resin was used as a control. The modulus of rupture for all groups of acrylic resin containing silica was significantly lower than for the control. The modulus of elasticity was not significantly greater than the control group. For the impact strength statistical analysis revealed a significant difference between the groups. There was a nonsignificant increase in the impact strength for specimens compared to the control. In conclusion the addition of silica to poly(methyl methacrylate) denture base materials did not produce a significant improvement in the transverse bend or impact strength compared to conventional heat-cured acrylic resin. The incorporation of untreated and surface treated silica cannot be recommended as a method of reinforcement. PMID- 16477119 TI - Bioperformance of shape memory alloy single crystals. AB - Shape memory alloys (SMA) represent a large family of alloys that show unique characteristics. They have been exploited in several fields for diverse applications. For the last 20 years, these alloys and more particularly Ni-Ti alloys have revolutionized the field of metallic biomaterials. Applications in the biomedical area are multiple and these materials improve significantly the quality of the diagnostics, treatments and surgeries. To our knowledge, most devices are made of SMAs in the polycrystalline form. Nevertheless, the single crystal form shows several promising advantages especially concerning its mechanical performances. In this paper we describe the advantages, advances and limits of using different SMA single crystals for biomedical applications, including biocompatibility and corrosion resistance. We also discuss the low response time of classical thermal SMAs as well as the new advances in research on magnetic SMA single crystals. PMID- 16477120 TI - Studies on quantitative analysis and automatic recognition of cell types of lung cancer. AB - Recognition of lung cancer cells is very important to the clinical diagnosis of lung cancer. In this paper we present a novel method to extract the structure characteristics of lung cancer cells and automatically recognize their types. Firstly soft mathematical morphology methods are used to enhance the grayscale image, to improve the definition of images, and to eliminate most of disturbance, noise and information of subordinate images, so the contour of target lung cancer cell and biological shape characteristic parameters can be extracted accurately. Then the minimum distance classifier is introduced to realize the automatic recognition of different types of lung cancer cells. A software system named "CANCER.LUNG" is established to demonstrate the efficiency of this method. The clinical experiments show that this method can accurately and objectively recognize the type of lung cancer cells, which can significantly improve the pathology research on the pathological changes of lung cancer and clinical assistant diagnoses. PMID- 16477122 TI - Fatigue and fracture in materials used for micro-scale biomedical components. AB - Some biomedical components involve the use of materials in microscopic quantities, i.e. in section sizes which are of the same order of magnitude as microstructural features in the material, such as grains. The mechanical behaviour of the material may be different when used in these quantities, compared to its behaviour in macroscopic amounts. An example of a microscopic component is the cardiovascular stent. To ensure the integrity of the stent during deployment and subsequent use, the designer must be able to simulate possible failure modes, i.e. monotonic fracture and fatigue, and the effect of stress concentrations. We carried out tests on specimens of 316L stainless steel, with and without stress concentrations. We found a significant size effect, in which the behaviour of these microscopic specimens was different from that of larger, macroscopic specimens. Microscopic specimens had lower tensile strengths and higher ductility. Under cyclic loading, the material's behaviour at large numbers of cycles was independent of specimen size, but the microscopic specimens were inferior at smaller numbers of cycles to failure. Fatigue limits for the notched specimens could be predicted using an existing theory (the Theory of Critical Distances) but parameter values were different at the macro- and micro scale. Thus, data from conventional, macroscopic specimens cannot be used to predict the behaviour of this material when used for microscopic components. Mechanical working and annealing strongly affected the tensile strength and ductility, but had no significant effect on fatigue behaviour. PMID- 16477121 TI - International harmonization of generic drugs: in vitro dissolution tests for Japanese and American generic tablets. AB - Ibuprofen tablets on the market in Japan and the USA were compared by manual- and automatic-dissolution tests according to USP24 criteria. Dissolution test were performed in 900 ml of phosphate buffer of pH 7.2 at 37.0+/-0.5 degrees C at 50 rpm for 60 min, and the time required for 70% dissolution (T70%) and 5% dissolution after 60 min (A60) were evaluated. The dissolution profiles of both Japanese and American tablets by the automatic-method showed almost the same profiles as those of the manual method. T70% of the American and Japanese tablets by the manual method were not significantly different (p>0.05) from the automatic method at various sampling positions. The A60 of the American and Japanese tablets by the manual-method was not significantly different (p>0.05) except at one position. The results indicate that the automatic-method was more reproducible than the manual-method, and also that systematic error was negligible. The T70% and A60 of the American tablets were significantly different (p<0.05) from the Japanese tablets. The American tablets were a film-coated over the-counter drug and the Japanese tablets were a sugar-coated prescription drug. There was a difference in dissolution behavior between the dosage forms of the two countries. PMID- 16477123 TI - The release of bovine serum albumin from polyurethane based hydrophilic and hydrophobic disks and microbiological interactions. AB - Polyurethane (PU) based hydrophobic and hydrophilic disks were prepared by graft copolymerization of PU with vinyl monomers, acryl amid (AAm), itaconic acid (IA), acrylic acid (AA) and crotonic acid (CA). Grafting reactions were performed in PU dioxane solution using benzoyl peroxide as initiator. Air-water and cyclohexan water contact angle values of the disks (thetaair and thetacy) were measured via "Captive Bubble Method". Surface free energy values of the disks were determined by using thetaair and thetacy via "Two Liquid Method" and those were compared with the swelling percentage values of the disks. The release of Bovine Serum Albumin (BSA) from protein added hydrophilic and hydrophobic PU disks were defined by spectrophotometric measurements at lambda=278 nm in phosphate buffer saline (PBS). BSA releasing is completed around 48 h for all disks. The disks were also characterized by FTIR spectra and SEM micrographs. In vitro attachment of Candida albicans (C. albicans) and Lactobacillus plantarum (L. plantarum) on PU based hydrophilic and hydrophobic disks was studied at 37 degrees C. While C. albicans attached on to all disks, L. plantarum only selectively attached. The attachment percentage values of C. albicans (25%) and L. plantarum (20%) on hydrophobic PU disk were higher than hydrophilic PU disks. The antimicrobial activity of disks was also tested against C. albicans and L. plantarum. IA and AAm grafted PU disks showed inhibitory effect for the both test bacteria. PMID- 16477124 TI - [Mechanisms of heavy metal cadmium tolerance in plants]. AB - Cadmium (Cd) is a strongly phytotoxic heavy metal, which inhibits plant growth and even leads to plant death. The main symptoms of Cd(2+) toxicity to plants are stunting and chlorosis. Plant has developed some functions for Cd(2+) tolerance, which include cell wall binding, chelation with phytochelatins (PCs), compartmentation of Cd(2+) in vacuole, and enrichment in leaf trichomes. However, Cd(2+) tolerance in plant is more likely involved in an integrated network of multiple response processes than several isolated functions cited above. In the network, the processes of sulfur metabolism, antioxidative response, and Cd(2+) transport across plasma and vacuole membrane in plant are closely related with Cd(2+) tolerance in plant. The processes of sulfur uptake, assimilation and sequential sulfur metabolism in plant respond to Cd(2+) stress. The expression of sulfur transporters with varied affinity was changed in different ways under Cd(2+) stress, and the high expression of ATP sulfurylase (APS) and adenosine 5' phosphosulfate reductase (APR), which may help to keep the supply of S(2-) for cysteine (Cys) synthesis. The efficiency of Cys synthesis may function in Cd(2+) detoxification, and the up-regulated expression of Ser acetyltransferase (SAT) and O-acetyl-ser (thiol)-lyase (OASTL) has been found in some Cd(2+) treated plants. Reduced glutathione (GSH) is an important antioxidant and the precursor of PCs, glutamylcysteine synthetase (GCS) and glutathione synthetase (GS) catalyze GSH synthesis from Cys, overexpression of the two enzymes can improve Cd(2+) tolerance in plant. PCs are more important Cd(2+) chelators than metallothioneins (MTs) in plants, and the expression of phytochelatin synthase (PCS) responds to Cd(2+) stress. Plant antioxidative system also contributes to Cd(2+) tolerance. The antioxidative response to Cd(2+)-induced oxidative stress varies in different plants and tissues and is also Cd(2+) concentration dependent, and the Cd hyperaccumulator plants show strong tolerance to oxidative stress. Some genes encoded metal transporters with Cd(2+) substrate specificity at plasma and vacuole membranes, which have been isolated and characterized in recent years. These genes play critical roles in Cd(2+) translocation, allocation, and compartmentation in plants. Despite the great progresses made in the field in recent years, there are still some issues which need further exploration, such as the detail of signal transduction and the responses of gene regulation to Cd(2+), the rhizosphere activation and root adsorption to soil Cd(2+), Cd(2+) trafficking in xylem and phloem, Cd(2+) translocation to fruit and seed, and the possible presence of a high-affinity Cd(2+) transporter in Cd hyperaccumulators. PMID- 16477125 TI - [Plant sulfate assimilation and regulation of the activity of related enzymes under cadmium stress]. AB - The complexation and sequestration of heavy metal ions (e.g. Cd) by the cysteine rich polypeptides known as phytochelatins (PC) are thought to confer heavy metal hyperaccumulation and tolerance in some plant species. PC is synthesized enzymatically from glutathione. The tripeptide glutathione is a product of primary sulfur metabolism. A variety of enzymes or proteins are involved in sulfur assimilation including sulfate transporters (STs), ATP sulfurylase (ATPS), APS reductase (APSR), sulfite reductase (SiR), glutathione synthetase (GS) and phytochelatin synthesis (PCS). These enzymes or proteins are upstream-regulated by Cd at either the metabolic or the genetic level under metal stress. Increasing evidence shows that enhancement of sulfate uptake and reduction occurs with the production of PC in plants under heavy metal stress. In this article, the key aspects of our recent understanding of regulatory mechanisms involved in the relation between the sulfate assimilation and phytochelatin synthesis are described. PMID- 16477126 TI - [Advances in cell biological researches on male sterility of higher plants]. AB - Male sterility of higher plants is multiform in pollen abortion and has varied and complicated mechanisms. It is an active field to probe the mechanisms. Recently, some new results in this field have been obtained by using the methods of cell biology, including the structure and function of tapetal cell, the changes in Ca(2+) distribution, ATPase activity distribution, cytoskeleton array and programmed cell death in anther cells. All of the results gave us some new understanding for the process of pollen abortion. These results will make a link between the researches of individual and molecular level in male sterility of higher plants, and help us understand the mechanisms of male sterility of higher plants. This paper summarizes the knowledge about aborting process of male sterile anther obtained by the methods of cell biology. PMID- 16477127 TI - Physiological and biochemical characteristics and molecular-biological identification of a gigantic tobacco mutant. AB - A new gigantic late-flowering tobacco mutant was isolated in tobacco field in Xunyang county, Shaanxi province, in 2001. It was rescued through tissue culture and a large amount of regenerated plantlets were obtained. The results of the comparison between the regenerated plants from this mutant and the wild type plant (K346) were as follows: (1) Morphological observation showed that the leaf number of the mutant was 3.3 times as many as that of the wild type, the height of mutant was 2.2 folds that of the wild type, and the mutant had the late flowering character. (2) Cytological examination showed that the mutant was a normal diploid, and the number of chloroplasts in a guard cell of mutant was 1.3 times that of the wild type. (3) Both chlorophyll a and b content of the mutant were larger than that of the wild type; soluble protein content of the mutant was 1.18 times as much as that of the wild type. Peroxidase isozyme and cytochrome oxidase isozyme electrophoresis analyses indicated differences between the mutant and the wild type. The soluble protein SDS-PAGE patterns showed the absence of four bands [P1 (114.6 kD), P2 (103 kD), P3 (66.2 kD), P4 (24 kD)] in the mutant. (4) RAPD analysis showed that the similarity index of the mutant and the wild type was 0.612. This suggested that there were changes at DNA level. The mRNA of mutant leaves at flowering stage was isolated; DDRT-PCR was carried out using 10 random primers, using OligodT(15)M (M=A/G/C) as anchor primer. It has been proved that gene expression at anthesis was different between the mutant and the wild type. (5) Genetic observation showed that the mutant was homozygotic and bred true. And the mutant was a late-flowering one. PMID- 16477128 TI - Fruit coloration difference between Fengwan, a late-maturing mutant and its original cultivar Fengjie72-1 of navel orange (Citrus sinensis Osbeck). AB - Fruit color is one of the most important external quality traits. Mutants with different color are useful for the study of regulating mechanism of coloration progress. A novel mutant, Fengwan, derived from Fengjie72-1 navel orange (Citrus sinensis L. Osbeck), has the distinctive characteristic of the coloration delay of 30 d or so, with the change in total soluble solid (TSS) and the ratio of TSS/acidity. In order to understand the mechanism underlying the difference of coloration between the mutant and the parental variety, the chlorophyll and carotenoid contents in the peel of two cultivars at different maturation stages were analyzed. The expression of genes responsible for some carotenoid biosynthetic enzymes (phytoene synthase, phytoene desaturase, zeta-carotene desaturase, beta-lycopene cyclase, beta-carotene hydroxylase) and chlorophyllase using the RT-PCR technique were also studied. The distinct decrease of chlorophyll in the peel of Fengwan navel orange occurred from early November to late November, about three weeks later than that of its original cultivar. Obvious accumulation of carotenoid in the peel of the mutant began on Dec.12, while that of the original cultivar began on Nov. 3. Analysis of independent samples t-test showed that the chlorophyll content of the peel of Fengwan navel orange from October to November was significantly higher, and the carotenoid content from December to January was significantly lower than that in the peel of the parental line, Fengjie72-1 navel orange. The expression of chlorophyllase gene in the peel of Fengwan navel orange reached a maximum in January and kept at a slightly lower level from October to December than that of Fengjie 72-1 orange, while the time of gene expression about some carotenoid biosynthetic enzymes reaching a maximum in the peel of the mutant were one month later than that of the original cultivar. PMID- 16477129 TI - [A preliminary analysis of the function of the OsRab5a gene in rice (Oryza sativa L.)]. AB - OsRab5a encodes a Rab5 protein in rice, which belongs to the Rab family of small GTP-binding protein superfamily. Transgenic plants with POsrab5a::beta glucuronidase (GUS) gene indicate that OsRab5a is expressed in callus, root, stem, leaf, root base and spikelet (Figs.1, 2H-1). Using the transgenic Arabidopsis with fused gene of GFP::OsRab5a and GFP::OsRab5aCA and by using FM4 64 (the lipophilic styryl dye) staining, we have demonstrated that OsRab5a is involved in vesicular transport and GFP-OsRab5a is localized on the plasma membrane and early endosomes (Fig.2A-C), while GFP-OsRab5aCA is localized in the cell membrane (Fig.2D-F). The reduction of expression of OsRab5a in callus RNA interference, however, led to the lethality of callus during differentiation (Fig.3D-F). The expression of OsRab5a was upregulated slightly by exogenous cytokinin (Fig.4), and was higher in the differentiation medium than that in selection medium (Fig.3G-I). These results suggest that OsRab5a plays an important role in callus differentiation probably through mediating hormone signal transduction. PMID- 16477130 TI - [Transformation of embryogenic Calli of Siberian wildrye grass (Elymus sibiricus L. cv. Chuancao No.2) mediated by agrobacterium]. AB - Formation of embryogenic calli of Siberian wildrye grass (Elymus sibiricus L. cv. Chuancao No.2) was induced from mature seeds as explants, and proliferated on MS medium containing 2,4-D 5.0 mg/L and KT 0.05 mg/L. An effective and stable callus regeneration system was established by optimizing the culture conditions (Tables 1, 2 and Fig.2). After the calli were subcultured 8 weeks, selected the whitish yellow-coloured compact nodular calli that transformed with plasmid pCAMBIA1304 carrying hygromycin resistance gene (hptII) and Pseudomonas pseudoalcaligenes insecticidal protein gene (ppIP), which was mediated by an Agrobacterium strain EHA105. Resistant plants were obtained after hygromycin selection (Figs.3, 4). Some important factors that affect the transformation efficiency were studied, which included selection pressure, time of embryogenic calli proliferation, OD value of Agrobacterium suspension, temperature, medium and time of co cultivation, and concentration of antibiotics used for suppressing the overgrowth of Agrobacterium in the course of transformation plant regeneration. This research is the first successful genetic transformation of Elymus sibiricus L. cv. Chuancao No.2 mediated by Agrobacterium. PMID- 16477131 TI - [Expression of an Arabidopsis actin-depolymerizing factor 4 gene (AtADF4) in tobacco causes morphological change of plants]. AB - The actin-depolymerizing factor 4 gene (ADF4) of Arabidopsis thaliana was cloned and sequenced (Figs.1, 2). The plant expression vector with ADF4 was constructed and transformed into tobacco by Agrobacterium tumefaciens. Molecular identification showed that the ADF4 gene was integrated into the genome of tobacco and expressed in transgenic tobaccos assayed by PCR and RT-PCR (Fig.3). Expression of an Arabidopsis thaliana ADF4 gene in tobacco resulted in morphological change of plants. The effects of ADF4 on transgenic tobaccos growth were as follows: the hypocotyls of transgenic plants were wavy, especially in darkness, whereas those of the control were straight (Fig.4A); the root hairs of transgenic plants were less than the control, and they were also wavy (Fig.4B); the parenchyma cells of transgenic plants were larger than the control and the arrangement of vascular bundle was out of order (Fig.4C); the flowering time of T(2) line was at least 7 days later than the control; the pollen tubes of transgenic plants were shorter than those of the control (Fig.4D). PMID- 16477132 TI - Over-expression of exotic superoxide dismutase gene MnSOD and increase in stress resistance in maize. AB - To evaluate the effect of over-expressed Mn-superoxide dismutase (MnSOD) on stress resistance in maize, an over-expression vector with MnSOD gene of wheat was constructed and used to transform embryonic calli of elite maize inbred line by gene gun. After screening with a concentration gradient of hygromycin, 9 plants were regenerated from the positive calli and reproduced fertile seeds, 5 of which were shown to be positive by PCR and Southern blot analysis. This result indicated that the exotic gene had been integrated in maize genome. SOD enzyme extracts were separated by non-denatured polyacrylamide gel electrophoresis of gradient concentration. After inhibition with 5 mmol/L H(2)O(2) to FeSOD and Cu/ZnSOD, MnSOD enzyme activity was assayed with nitrotetrazolium chloride blue staining. All the 5 plants shown to be positive in Southern blot had higher MnSOD activity than untransformed control. Electric conductivity of leaf exudate was assayed after damage treatment with methyl viologen. The result suggested that the resistance of the transformed plants to oxidative damage was significantly higher than untransformed control. PMID- 16477133 TI - Cloning and molecular characteristic of the metalloprotease gene LeftsH6 from tomato. AB - The full-length 2213-bp ftsH (filamentation temperature-sensitive H) cDNA was cloned from the cDNA library of heat-shocked tomato leaves. According to an open reading frame of 2019-bp, the deduced protein precursor was predicted to target chloroplast. The putative AAA (ATPases associated with diverse cellular activities) domain and the Zn(2+)-binding domain, characteristic of FtsH metalloproteases family, were found in the FtsH-like protein. Most similar to the FtsH6 of Arabidopsis thaliana, the tomato ftsH-like gene was named as Lycopersicon esculentum filamentation temperature-sensitive H6 (LeftsH6). Purified FtsH degraded casein but not BSA in vitro, whereas a FtsH mutant with the Glu(472) in the zinc-binding motif replaced by Gln had lost the protease activity. A single copy of LeftsH6 was detected in tomato genome by Southern blot analysis. Northern and Western blot analyses revealed consistently the heat inducible character of the LeftsH6 gene. No LeftsH6 expression was detected after cold, salt, drought or light stress. The results provided the first experimental evidence of the existence of heat-inducible ftsH gene in higher plants. PMID- 16477135 TI - Cloning, characterization and expression analysis of the receptor-like kinase gene (RLK) in common wheat. AB - To investigate whether homologs of wheat (Triticum aestivum L.) LRK10 gene was expressed in powdery mildew-resistant lines after inoculation with Blumeria graminis f. sp. tritici, one degenerate primer for 5'-RACE was designed according to the 6th kinase subdomain of LRK10 and other plant kinases. 5'-RACE was performed with the template cDNA synthesized with RNA extracted from seedling leaves of a powdery mildew-resistant wheat line "99-2439" after inoculation with B. graminis. One 1551 bp cDNA fragment representing a protein kinase gene was obtained (S1125, GenBank accession number: AY584533). Subsequently, a 2255-bp full-length cDNA clone with a complete encoding region (open reading frame, ORF) was obtained by RACE. The clone encodes a polypeptide consisting of 637 amino acid residues. The result of homology search showed that it belongs to a receptor like kinase gene family in wheat, which was named as wlrk (wheat leaf rust kinase) previously. Similar to LRK10, this protein kinase has five distinct domains: a hydrophobic signal sequence at the amino-terminus, a putative extracellular domain, a transmembrane domain, a highly charged sequence and a serine/threonine kinase domain at the carboxy-terminus, and thus it was named as TaLRK (Triticum aestivum LRK). The expression pattern of TaLRK at transcription level in leaves after B. graminis infection was investigated by semi-quantitative RT-PCR (semi-QRT-PCR), using wheat actin gene as a control. The result showed that the transcription of TaLRK was significantly enhanced by B. graminis infection. The expression pattern of TaLRK in different tissues showed that this new wheat RLK gene was expressed only in green parts of wheat. This study suggests that TaLRK may function in wheat powdery mildew resistance responses. PMID- 16477134 TI - Silicate improves growth and ion absorption and distribution in aloe vera under salt stress. AB - Si 2.0 mmol/L in irrigation solution alleviated significantly the inhibition of NaCl stress of 100 or 200 mmol/L to aloe growth. Exogenously applied Si decreased significantly Na(+) and Cl(-) contents, increased K(+) content and K(+)/Na(+) ratio and selectivity ratio of absorption (AS(K, Na)) and of translocation (TS(K, Na)) to K(+) and Na(+) in aloe plant under both NaCl 100 and 200 mmol/L stresses for 30 d. In this way, the ion homeostasis in aloe plant under NaCl stress was maintained, as was proved by X-ray microanalysis of root tip and leaf across sections. One of the mechanisms to achieve this may be the significant enhancement of H(+)-ATPase activities by the addition of silicate in plasma membrane and tonoplast, H(+)-pyrophosphatase (H(+)-PPase) activity in tonoplast isolated from aloe root tips under NaCl stress. PMID- 16477136 TI - [Choline chloride protects cell membrane and the photosynthetic apparatus in cucumber seedling leaves at low temperature and weak light]. AB - Choline chloride 1.07 mmol/L treatment diminished the saturated lipid contents of the fatty acid components mainly the phosphatidylglycerol (PG) resulting in the decrease of the saturation of lipid (Table 1), declined the permeability of cell membrane and the production of MDA from lipid peroxidation (Fig.2) in the cucumber seedling leaves under low temperature and weak light (6 degrees C, PFD 100 micromol m(-2) s(-1)). Furthermore, the choline chloride treatment alleviated the degradation of chlorophyll pigments especially chlorophyll b, the decrease in maximum photochemical efficiency (Fv/Fm), the capture efficiency of excited energy (Fv'/Fm'), the photochemical quenching coefficient (q(p)) and the actual photochemical efficiency (Phi PSII) of PSII (Table 2, Fig.3A, B & C), and decreases in activity of antioxidant enzymes such as POD, APX and CAT (Fig.4) in chilled leaves under weak light. In addition, choline chloride treatment increased the non-photochemical quenching (NPQ) (Fig.3D) and the proline content (Fig.5) in chilled leaves under weak light. The above results indicate that choline chloride protected the cell membrane and the photosynthetic apparatus in cucumber seedling leaves from chilling stress in weak light. PMID- 16477137 TI - [Effects of NaCl stress on cation contents in seedlings of two pumpkin varieties]. AB - Effects of different concentration NaCl stress on cation contents of pumpkin cultivars (Cucurbita ficifolia and Cucurbita moschata) usually used as rootstocks of cucumber were studied. The results showed that, in both cultivars treated with 150, 300 and 500 mmol/L NaCl for 7 d, increasing salinity caused in increasing retardation of shoot and root growth, and reduction of shoot relative water content (Table 1). With increasing NaCl concentration, Na(+) content increased while K(+) content decreased distinctly in roots, stems and leaves of both cultivars. And Na(+) and K(+) contents in different organs were in the orders roots>stems>leaves and stems>leaves>roots respectively (Fig.3). There were less Na(+) in root, stem and leaf (Fig.3), more free proline and soluble sugar in leaves (Fig.1, 2), and higher shoot relative water content (Table 1) in C. ficifolia than in C. moschata. So there are differences between C. ficifolia and C. moschata in osmotic adjustment mechanisms and selective absorption and transportation of different cations, and the salt tolerance of C. ficifolia is higher than C. moschata. PMID- 16477138 TI - [Expression characteristics of MtPAP1 and its exotic expression in Arabidopsis affecting organic phosphorus absorption of plants]. AB - The MtPAP1 encodes 465 amino acids (Fig.1). The transcript of MtPAP1 was detected mainly in leaf under high-phosphate conditions (Fig.2). While under low-phosphate condition, the transcript level was low in leaves and very high in roots, with the strongest hybridization signal detected in roots (Fig.2). Chimeric MtPAP1 binary vector was constructed and the transcript levels of MtPAP1 in roots of transgenic T(3) Arabidopsis lines were analyzed (Fig.3). Under the condition in which phytate was the sole source of phosphorus, transgenic Arabidopsis plants over-expressing MtPAP1 showed acid phosphatase activities in root apoplast increasing markedly than that of the control plant (Fig.4). The results of analysis of organic phosphorus degradation in liquid culture by HPLC indicated that the APase secreted by the transgenic plants could quickly degrade the phytate (Fig.5). Significant increase in biomass production, Pi and total phosphorus content of plants achieved in the transgenic lines when phytate, an organic phosphorus compound, was supplied as the sole source of phosphorus (Fig.6,7). PMID- 16477139 TI - [Effects of exogenous silicon on active oxygen scavenging systems in chloroplasts of cucumber (Cucumis sativus L.) seedlings under salt stress]. AB - With K(2)SiO(4) (1.0 mmol/L) treatment, the effects of Si on the distribution of Na(+), K(+) to chloroplasts and antioxidant system of cucumber leaves under 50 mmol/L NaCl stress were studied. The results showed that there was a selective transport of K(+) into the chloroplasts so that Na(+) content of chloroplasts was lower under Si treatment (Table 1); H(2)O(2) and MDA contents in chloroplasts were significantly decreased (Fig.1), and the activities of SOD, APX, GR and DHAR were increased simultaneity (Fig.2), and AsA, GSH contents were also increased in chloroplasts of salt-stressed cucumber by additional Si treatment (Fig.3). It may be concluded that Si could decrease absorption of Na(+) and increase ability of active oxygen scavenging in chloroplasts, therefore the injury of chloroplast membrane under salinity stress in cucumber was alleviated. PMID- 16477140 TI - [The distribution of ATPase in developmental anther of rice]. AB - The distribution of ATPase was studied using lead precipitation technique during anther development in rice. The ATPase reactive precipitates (ppts) were located mostly in the nucleus of microspore mother cells (MMC) and only a few in the cytoplasm (Plate I-1). Anther wall had differentiated into four layers of cells and a few precipitates were located in the cells except the nucleus of tapetal cells where there were many ATPase reactive precipitates (Plate I-2). After meiosis of MMC, tapetal cells formed many endoplasmic reticula in its cytoplasm but still contained a few ppts. In the cells of epidermis, endothelium and middle layer, the ppts increased evidently in plasma membrane and near cytoplasm than before (Plate I-5). There were a large number of ppts located in the pollen wall during pollen development (Plate I-6), suggesting that ATPase is necessary for the construction of pollen wall. The exine of pollen wall of rice was constructed during microspore development and consisted of sporopollenin which came from tapetal cells. The ppts in exine also came from tapetal cell (Plate II-7). The intine of pollen wall was constructed during the stage of 2-cellular pollen and consisted of cellulose material coming from vegetative cell of pollen. The ATPase and ppts in intine came from vegetative cell of pollen (Plate III-7). Vegetative cell contained more ppts than generative cell during the development of 2 cellular pollen (Plate II-4, 5). The amount of ppts between two sperm cells in a pollen grain was also different (Plate IV-3,4). The physiological functions of ATPase located in different cells and different parts in the cells during anther development of rice were analyzed. PMID- 16477141 TI - Na+/K+ selectivity of leaf sheath in wheat cultivars differing in salt tolerance. AB - A salt-tolerant wheat cultivar (DK961) and a salt-sensitive one (LM15) were exposed to 200 mmol/L NaCl for 3 d to study selectivity in transport of K(+) and Na(+) of the leaf sheath. Na(+) content of leaf sheath and K(+) content of leaf blade of DK961 were significantly higher than those of LM15, which led to a lower Na(+)/K(+) ratio of leaf blade of DK961 than that of LM15. These results suggested that leaf sheath of DK961 had much stronger abilities of limiting Na(+) transport and allowing K(+) transport from leaf sheath to leaf blade than that of LM15 to maintain lower Na(+)/K(+) ratio of leaf blade under NaCl stress, indicating that leaf sheath may play an important role in wheat salt tolerance. PMID- 16477142 TI - Krox-20 gene expression: influencing hindbrain-craniofacial developmental interactions. AB - Krox-20 is a C(2)H(2)-type zinc-finger transcription factor that plays an essential role in hindbrain development. The Krox-20 null mutation results in hindbrain anomalies that result in neonatal death due to respiratory and feeding deficits. Here we review our studies of how the Krox- 20 null mutation impacts the development of motor and sensory systems critical for the production of consummatory behaviors (suckling/chewing). First, we demonstrated that Krox-20 null mutants suffer a selective loss of primary jaw-opening muscles during prenatal development. In vivo and in vitro studies are reviewed that highlight intrinsic defects in mutant jaw-opener muscles that contribute to muscle degeneration. Next we focus on the impact of the mutation on proprioceptive neurons activated during consummatory behaviors. Mesencephalic trigeminal (Me5) neurons are primary sensory neurons that relay jaw proprioception to the central nervous system. These cells are unique because their cell bodies are located in the central as opposed to the peripheral nervous system. Data are reviewed that demonstrate the impact of the mutation on Me5 neurons, a cell group traditionally thought to emerge from the mesencephalon. We show that Krox-20 null mutants have twice as many Me5 neurons relative to wildtypes at E15, but by birth have half the number of Me5 cells as wildtypes. TUNEL assays performed in each set of studies reveal that Krox-20 expression acts to protect both muscle and mesencephalic trigeminal neurons against apoptosis, suggesting that Krox-20, in addition to its role in hindbrain patterning, has a broader, long-lasting role in development. PMID- 16477143 TI - Expression of synaptotagmin 1 in the taste buds of rat gustatory papillae. AB - Synapses between taste receptor cells and primary sensory afferent fibers transmit the output signal from taste buds to the central nervous system. The synaptic vesicle cycle at the synapses involves vesicle docking, priming, fusion, endocytosis, and recycling. Many kinds of synaptic vesicle proteins participate in synaptic vesicle cycles. One of these, synaptotagmin 1, binds Ca(2+) phospholipids with high affinity and plays a role in Ca(2+) regulated neurotransmitter release in the central and peripheral nervous systems. However, the expression patterns of synaptotagmin 1 in rat taste tissues have not been determined. We therefore examined the expression patterns of synaptotagmin 1 and several cell specific markers of type II and III cells in rat taste buds. RT-PCR assay showed that synaptotagmin 1 mRNA was expressed in circumvallate papillae. In fungiform, foliate, and circumvallate papillae, the antibody against synaptotagmin 1 yielded the labeling of a subset of taste bud cells and intra- and subgemmal nerve processes. Double labeled experiments showed that synaptotagmin 1 positive cells co-expressed type III cell markers, PGP 9.5, and NCAM. Intragemmal nerve processes positive for synaptotagmin 1 co-expressed PGP 9.5. Conversely, all synaptotagmin 1 expressing cells did not co-expressed type II cell markers, PLCbeta2, or gustducin. These results show that synaptotagmin 1 may play some regulatory roles in vesicle membrane fusion events with the plasma membrane at the synapses of type III cells in rat taste buds. PMID- 16477144 TI - Jacalin and peanut agglutinin (PNA) bindings in the taste bud cells of the rat: new reliable markers for type IV cells of the rat taste buds. AB - Lectin histochemistry of Jacalin (Artocarpus integrifolia) and peanut agglutinin (PNA), specific lectins for galactosyl (beta-1, 3) N-acetylgalactosamine (galactosyl (beta-1, 3) GalNAc), was applied to the gustatory epithelium of the adult rat. In the ordinary lingual epithelium, Jacalin and PNA labeled the cell membrane from the basal to granular cell layer. They also bound membranes of rounded-cells at the basal portion of taste buds, but the number of PNA labeled cells was smaller than that of Jacalin labeled cells. There was no apparent difference in the binding patterns of Jacalin and PNA among the taste buds of the lingual papillae and those of the palatal epithelium. Occasionally, a few spindle shaped cells were labeled with Jacalin, but not with PNA. Double labeling of Jacalin and alpha-gustducin, a specific marker for type II cells, revealed that Jacalin-labeled spindle-shaped taste cells were immunonegative for alpha gustducin. Spindle-shaped cells expressing protein gene product 9.5 (PGP 9.5) immunoreactivity lacked Jacalin labeling. During the development of taste buds in circumvallate papillae, the binding pattern of Jacalin became almost identical from postnatal day 5. The present results indicate that rounded cells at the basal portion of the taste buds cells (type IV cells) bind to Jacalin and PNA, and these lectins are specific markers for type IV cells of the rat taste cells. PMID- 16477145 TI - The odontoblast as a sensory receptor cell? The expression of TRPV1 (VR-1) channels. AB - Previous reports have shown the expression of several mechanosensitive ionic channels on the plasma membrane in odontoblasts, which are the cells responsible for dentin formation. The membrane characteristics of odontoblasts imply that they could play critical roles in the mechano-transduction of fluid displacement within dentinal tubules into the electrical cell signals, to carry dentin sensation to the central nervous system. However, the direct ionic mechanism underlying such a dentin nociceptive function remains unclear. In the present study, we investigated the expression of the transient receptor potential vanilloid subfamily member 1 (TRPV1) channel--which essentially contributes to the detection of pain sensation--in rat odontoblasts by immunohistochemical and nystatin perforated patch-clamp techniques. Immunohistochemical observation showed the localization of TRPV1-immunoreactions on the distal regions of odontoblast membranes. In the patch-clamp experiments, we observed capsaicin induced inward currents that were inhibited by capsazepine, a TRPV1 channel antagonist. Our results indicate a significant expression of TRPV1 channels in odontoblasts, suggesting that odontoblasts may directly respond to noxious stimuli such as a thermal-heat stimulus, and point to the necessity for a reconsideration of the cellular mechanisms of dentin sensation based on the transmembrane ionic signals in odontoblasts. PMID- 16477146 TI - Immunocytochemical localization of the neurokinin 1 receptor in rat dental pulp. AB - The dentin-pulp complex is a peripheral end-organ supplied by dense sensory nerve fibers. Substance P, a representative neuropeptide widely distributed in the dental pulp, has been reported to play roles in pain transmission and the amplification of inflammation. We analyzed here the expression of the neurokinin 1 (NK1) receptor, preferentially activated by substance P, using immunocytochemistry in rat dental pulp at both the light and electron microscopic levels. Conspicuous NK1 receptor immunoreactivity was found in the odontoblasts; immunolabelings were present at their plasma membrane and endosomal structures, especially in their cytoplasmic processes. Immunoreactions for NK1 receptor were also detectable in a part of the nerve terminals associated with the cytoplasmic processes of the odontoblasts. Furthermore, the endothelial cells of capillaries and post-capillary venules and the fibroblasts were labeled with the NK1 receptor in the subodontoblast layer. These findings suggest that pulpal cells and nerve fibers are targets for substance P that mediate multiple functions, including a vasoactive function and the regulation of vascular permeability as well as the modulation of pain transmission. PMID- 16477147 TI - Neurotrophin-4/5-depletion induces a delay in maturation of the periodontal Ruffini endings in mice. AB - Neurotrophin-4/5 (NT-4/5) - a member of the neurotrophic factors - is a ligand for TrkB, which has been reported to be expressed in the mechanoreceptive Ruffini endings of the periodontal ligament. The present study examined developmental changes in the terminal morphology and neural density in homozygous mice with a targeted disruption of the nt-4/5 gene and wild-type mice by immunohistochemistry for protein gene product 9.5 (PGP 9.5), a general neuronal marker, and by quantitative analysis using an image analyzer. Postnatal development of terminal Schwann cells was also investigated by enzymatic histochemistry for non-specific cholinesterase activity (ChE). Furthermore, the immuno-expression of TrkB and low affinity nerve growth factor receptor (p75-NGFR) was surveyed in the periodontal Ruffini endings as well as trigeminal ganglion. At postnatal 1 week, the lingual periodontal ligament of both types of mice contained PGP 9.5-positive nerve fibers showing a tree-like ramification with axonal swellings in their course. In both types of mice at 2 weeks of age, comparatively thick nerve fibers with a smooth outline increased in number, and frequently ramified to form nerve terminals with dendritic profiles. However, no typical Ruffini endings with irregular outlines observed in the adult wild-type mice were found in the periodontal ligament at this stage. At postnatal 3 weeks, typical Ruffini endings with irregular outlines were discernable in the periodontal ligament of the wild type mice while the dendritic endings showing smooth outlines were restricted to the homozygous mice. After postnatal 8 weeks, both types of mice showed an increase in the number of Ruffini endings, but the morphology differed between the wild-type and NT-4/5 homozygous mice. In the wild-type mice, a major population of the Ruffini endings expanded their axonal branches and developed many microprojections, resulting in a reduction of endings with smooth outlines. In contrast, we failed to find such typical Ruffini endings in the periodontal ligament of the homozygous mice: A majority of the periodontal Ruffini endings continued to show smooth outlines at postnatal 12 weeks. Quantitative analysis on neural density demonstrated a reduction in the homozygous mice with a significant difference by postnatal 8 weeks. Enzymatic histochemistry for non-specific ChE did not exhibit a distinct difference in the distribution and density of terminal Schwann cells between wild-type and homozygous mice. Furthermore, TrkB and p75 NGFR mRNA and proteins did not differ in the trigeminal ganglion between the two types. The periodontal Ruffini endings also displayed immunoreactivities for TrkB and p75- NGFR in both phenotypes. These findings suggest that the nt-4/5 gene depletion caused a delay in the formation and maturation of the periodontal Ruffini endings in the mice by inhibiting the growth of the periodontal nerves at an early stage, and indicate that multiple neurotrophins such as NT- 4/5 and BDNF might play roles in the development and/or maturation of the periodontal Ruffini endings. PMID- 16477148 TI - Requirement of occlusal force for maintenance of the terminal morphology of the periodontal Ruffini endings. AB - The present study examined whether mechanical stimulation is required for morphological maintenance of the Ruffini endings--primary mechanoreceptors in the periodontal ligament of the rat incisors, using a hypofunctional model by immunohistochemistry for protein gene product 9.5. The periodontal Ruffini endings of adult rats were observed to be restricted to the alveolar half of the lingual ligament where they displayed a dendritic arborization of expanded axon terminals with threadlike microprojections. In the experimental group, the tips of the upper and lower incisors were unilaterally ground to reduce mechanical stimulation of the ligament, i.e. occlusal force. A reduction in the occlusal force induced morphological changes in the terminal morphology of the periodontal Ruffini endings: they became smooth, unlike the irregular profiles exclusively observed in the control group. Quantitative analysis demonstrated significantly lower percentages of immunoreactive areas in the restricted portion on the ground sides than in normal animals. When incisor occlusion was re-established, the terminal portions of the Ruffini endings returned to their normal appearance, and the percentages of immunoreactive areas also recovered. The present results confirm the reduced size and number of axon terminals of periodontal Ruffini endings following reduced occlusal force and restoration of the morphological alteration after the re-establishment of incisor occlusion, indicating that proper mechanical stimulation is an important factor for maintaining the morphology of mechanoreceptors. PMID- 16477149 TI - Activation of the caspase cascade underlies the rat trigeminal primary neuronal apoptosis induced by neonatal capsaicin administration. AB - The systemic administration of capsaicin is known to cause a massive loss of sensory primary neurons in newborn rats. Here we examined the trigeminal ganglion neurons immunohistochemically for the possible induction of activated forms of caspases-9 and -3 following a subcutaneous injection of capsaicin in newborn rats. The DNA fragmentation signal was labeled by a TUNEL method. TUNEL-positive neurons were rare (< 0.5%) at 24 h after injection of the vehicle without capsaicin. After the capsaicin injection, TUNEL-positive neurons began to increase by 12 h, reached a peak at 24 h (11.4%), and returned to the control level by 120 h. Vehicle control levels of caspase- 9-immunoreactive (ir) and caspase-3-ir neurons were low (< 0.5%). Neonatal capsaicin administration induced caspase-9-immunoreactivity (ir) and -3-ir. The temporal distributions of caspase 9-ir and caspase-3-ir neurons were similar to those of TUNEL-positive neurons with peak expressions at 24 h of 13.2 and 11.1%, respectively. A double-stain analysis at 24 h post-injection indicated 72% of TUNEL-positive neurons were caspase-9-ir, and 70% caspase-3-ir. Conversely, 78 and 68% of caspase-9-ir and caspase-3-ir neurons, respectively, were TUNEL-positive. Comparison of two adjacent sections immunostained for the two different antigens revealed the co expression of the two caspases. These results suggest that neonatal capsaicin triggers the caspase cascade and, thereby, induces trigeminal primary neuronal apoptosis. PMID- 16477151 TI - Prenatal development of NMDA receptor composition and function in trigeminal neurons. AB - The prenatal development of neural circuits for rhythmical oral-motor behaviors used for feeding is essential for the survival of the newborn mammal. The N methyl-D-aspartate (NMDA) receptor plays a critical role in brainstem circuits underlying postnatal oral-motor behaviors. To understand a role for the NMDA receptor in the emergence of sucking behavior we conducted physiological and immunohistochemical experiments using fetal rats. Physiology experiments examined the development of the NMDA dose response of the brainstem circuit responsible for generating rhythmical trigeminal activity by recording trigeminal motor outputs using an in vitro preparation. The high dose of NMDA agonist bath application affected the mean cycle duration of rhythmical trigeminal activity (RTA) at both embryonic day (E) 18-19 and E20-21 in comparison with standard concentration of NMDA agonist. NMDA receptor immunohistochemistry studies, using antibodies directed against subunits NR1, NR2A, NR2B, NR3A and NR3B were performed to determine the prenatal regulation of NMDA subunits in trigeminal motoneurons (Mo5), and mesencephalic trigeminal neurons (Me5) between E17 to E20. In Mo5, NR1, NR2A, NR2B and NR3A immunoreactivity was observed throughout the time frame sampled. NR3B immunoreactivity was not observed in Mo5 or Me5. In Mo5, there was a significant decrease in the percentage of NR2B immunoreactive neurons between E17 and E20, and a concurrent increase in the NR2A/NR2B ratio between E17 and E20. In Me5, NR1, NR2A and NR3A immunoreactivity was observed throughout the time frame sampled; a significant decrease in the percentage of NR2A immunoreactive neurons between E17 and E20, and NR3A immunoreactive neurons between E17 and E18 occurred. The timing of subunit changes between E17 and E18 is coincident with the prenatal emergence of rhythmical jaw movements, and in vitro rhythmical trigeminal activity, shown in earlier studies. Our data suggest that NMDA receptor plays an important role in the development and function of prenatal oral-motor circuits. PMID- 16477150 TI - Postnatal development of substance P-immunoreaction in the trigeminal caudalis of neonatally capsaicin-treated mice. AB - The trigeminal subnucleus caudalis (Vc) is a critical relay site for processing nociceptive afferent input from the orofacial area in addition to its modulation by neuroplastic change. Although an administration of capsaicin in neonates induces a selective destruction of substance P (SP)-immunoreactive nerve fibers, little information is available regarding its detailed effects on the Vc, particularly during postnatal development. The present study examined postnatal changes in the distribution of SP in the Vc and trigeminal ganglion (TG) by immunohistochemical techniques in naive (NV) and neonatally capsaicin-treated (CP) mice, combined with a quantitative analysis. The neonatal mice received a single subcutaneous injection of capsaicin (50 mg/kg) at 48 hours after birth. The neural density of the SP-immunoreaction decreased to approximately a quarter of that in 1-week-old NV mice but increased to three-quarters of that in the NV in the superficial area after postnatal week 2. A double staining with SP and myelin basic protein confirmed the absence of any SP-immunoreaction in the myelinated nerve fibers in both NV and CP mice. The SP-immunoreaction never overlapped with non-peptidergic IB4-labeled neurons in the Vc and TG of either group. Neither the size distribution of SP-positive neurons nor their relative ratio in the TG differed between NV and CP mice at the ages of postnatal weeks 1 and 8. These findings indicate two putative origins for the emergent SP immunoreaction in the superficial layer of the Vc of the CP mice: the surviving trigeminal neurons with SP against capsaicin treatment and/or intrinsic neurons/interneurons in the Vc without SP under normal conditions. PMID- 16477152 TI - Needs assessment for continuing education activity. PMID- 16477153 TI - What is the "purple heel"? PMID- 16477158 TI - Offloading diabetic foot ulcers. PMID- 16477159 TI - Identifying infection in chronic wounds. PMID- 16477160 TI - Preparation and use of a scabies skin scraping kit. PMID- 16477161 TI - 2006 Medicare funding assigned for wound care services. PMID- 16477162 TI - 20 years of wound care: where we have been, where we are going. PMID- 16477163 TI - A look back at the 20th Anniversary Symposium. PMID- 16477164 TI - Chronic venous insufficiency in HIV-positive persons with and without a history of injection drug use. AB - OBJECTIVE: To examine chronic venous insufficiency in human immunodeficiency virus-positive persons with and without a history of injection drug use and to examine the extent to which neuropathy further increased the risk of chronic venous insufficiency. DESIGN: Cross-sectional stratified design with quota sampling. SETTING: Infectious diseases clinic in a large, urban midwestern city. PARTICIPANTS: Human immunodeficiency virus-positive persons, 27 with no history of injection drug use and 46 with a history of injection drug use, who met the inclusion criteria, including being 30 to 65 years of age, not pregnant, and willing to respond to a questionnaire and have their lower legs examined, were enrolled until the quota for each stratum (no injection drug use and injection drug use) was filled. MAIN OUTCOME MEASURES: Chronic venous insufficiency clinical classification, injection drug use history, and presence of peripheral neuropathy. MAIN RESULTS: Sixty-one percent of injection drug users (28/46) presented with severe chronic venous insufficiency compared with 11% (3/27) of noninjection drug users (P< .001). The presence of lower extremity neuropathy was not significantly related to chronic venous insufficiency classification. CONCLUSIONS: This is the first study to report the high risk of chronic venous insufficiency in human immunodeficiency virus-infected persons who inject drugs. Chronic venous insufficiency should be assessed in human immunodeficiency virus positive persons when there is a history of injection drug use, and measures to protect the legs should be implemented. PMID- 16477165 TI - The role of nutrition in wound care. AB - PURPOSE: To provide the physician and registered professional nurse with an understanding of how nutrition impacts wound healing. TARGET AUDIENCE: This continuing education activity is intended for physicians and nurses with an interest in learning more about the role nutrition plays in wound healing and how to supplement a patient's diet according to individual needs. OBJECTIVES: After reading the article and taking the test, the participant should be able to: 1. Determine a patient's daily caloric requirements.2. Identify signs of dehydration and malnutrition. 3. Understand the role specific vitamins and minerals play in the wound healing process. PMID- 16477168 TI - CD36: taste the difference? PMID- 16477169 TI - An update on parenteral lipids and immune function: only smoke, or is there any fire? AB - PURPOSE OF REVIEW: This paper synthesizes information from recent studies on the modulation of immune responses by lipid emulsions that are applied as part of parenteral nutrition. This issue is especially relevant in light of the high rate of infectious complications and disturbed inflammatory responses in patients receiving this form of nutritional support. RECENT FINDINGS: Studies reporting on novel emulsions based on olive and fish oils, structured lipids or mixed-type emulsions in which various lipid species replace conventional long-chain triglycerides indicate that these lipids are generally well tolerated. While long chain triglycerides may promote inflammation due to conversion of n-6 polyunsaturated fatty acids into arachidonic acid-derived eicosanoids, structured lipids and olive oil emulsions appear more immune-neutral. Leukocyte-activating effects of medium-chain triglycerides in experimental studies await further characterization in vivo. A body of evidence shows that immune modulation by fish oil emulsions is essentially anti-inflammatory in nature. This is in line with the observation that n-3 polyunsaturated fatty acids in fish oil replace arachidonic acid in cell membranes as an eicosanoid substrate, resulting in a decreased production of pro-inflammatory mediators. Importantly, recent investigations indicate beneficial effects of parenteral fish oil on relevant clinical outcome measures. SUMMARY: The characteristics of, and mechanisms behind, the effects of various parenteral lipids on immune function are becoming increasingly well understood. The practical relevance of many of these findings is not immediately clear, however, and will have to be substantiated in adequately powered trials before we can translate these findings into a tailored approach for specific clinical situations. PMID- 16477170 TI - Stearoyl-coenzyme A desaturase 1, sterol regulatory element binding protein-1c and peroxisome proliferator-activated receptor-alpha: independent and interactive roles in the regulation of lipid metabolism. AB - PURPOSE OF REVIEW: With the increasing incidence of obesity today, related complications such as diabetes, insulin resistance and hepatic steatosis are also becoming major concerns. Since these conditions share a common factor, aberrations in lipid metabolism, understanding the molecular changes that lead to abnormal lipid partitioning has become key to combating the obesity epidemic. RECENT FINDINGS: The enzyme stearoyl-coenzyme A desaturase 1 (SCD1) has been shown to be intimately involved in both the lipogenic as well as the lipid oxidative pathways. Our studies with the SCD1 mouse model have established that these animals are lean and protected from leptin deficiency-induced and diet induced obesity. Consequently, they also show greater whole body insulin sensitivity than wild-type mice. SCD1 mice have decreased expression of genes of lipogenesis and increased expression of lipid oxidative genes. The main transcription factors controlling genes of lipid synthesis and oxidation are sterol regulatory element binding protein-1c and peroxisome proliferator activated receptor-alpha (PPARalpha), respectively. Here, we review some studies that show that the effects of SCD1 deficiency on whole body adiposity may be partly dependent on sterol regulatory element binding protein-1c, but are most likely independent of peroxisome proliferator-activated receptor-alpha. SUMMARY: Our findings indicate that SCD1 is a key controller of lipid partitioning between lipogenesis and oxidation. While some questions regarding the molecular changes downstream of SCD1 deletion are yet to be answered, the studies outlined below clearly point to SCD1 as a highly promising target in combating obesity as well as related complications. PMID- 16477171 TI - Cyclooxygenase-2 inhibitors: what went wrong? AB - PURPOSE OF REVIEW: This review describes the biology underpinning the development of selective cyclooxygenase-2 inhibitors, documenting the clinical experience from the pivotal gastrointestinal safety trials to their eventual withdrawal or labelling with cardiovascular safety warnings. RECENT FINDINGS: The elucidation of differences between the active sites of cyclooxygenase-1 and cyclooxygenase-2 allowed the targeted design of the selective cyclooxygenase-2 inhibitors known as coxibs. These were developed and marketed as non-steroidal anti-inflammatory drugs (NSAIDs) that had improved upper gastrointestinal safety compared with older non-selective NSAIDs. A large-scale study with arthritis patients to evaluate upper gastrointestinal safety, however, demonstrated that celecoxib was not superior in terms of upper gastrointestinal safety compared with the older non-selective NSAIDs that were used as comparators. In an equally large study with arthritis patients, a more selective cyclooxygenase-2 inhibitor, rofecoxib, did have improved upper gastrointestinal safety compared with the non-selective non-steroidal anti-inflammatory drug naproxen. Although concomitant clinical trial evidence emerged that rofecoxib increased cardiovascular risk, this was discounted by its pharmaceutical company owner. Despite the lack of improved upper gastrointestinal safety with celecoxib and the evidence of cardiovascular risk with rofecoxib, both agents had widespread clinical use for 4-5 years. This was not diminished by the publication of plausible eicosanoid-based biological mechanisms whereby selective cyclooxygenase-2 inhibition could increase cardiovascular risk. Finally, clinical trials involving patients with colorectal cancer and post-operative pain revealed increased cardiovascular risk with all members of this class of drug. SUMMARY: These events provide a case study of a failure of the medical journal literature to guide drug usage. PMID- 16477172 TI - The independent effects of eicosapentaenoic acid and docosahexaenoic acid on cardiovascular risk factors in humans. AB - PURPOSE OF REVIEW: This review details the independent effects of purified eicosapentaenoic acid and docosahexaenoic acid on cardiovascular risk factors in humans. We report data from the recent literature and our own controlled clinical trials which compared the independent effects of these fatty acids in individuals at increased risk of cardiovascular disease, namely overweight hyperlipidaemic men and treated-hypertensive, type 2 diabetic men and women. We discuss the biological effects of these fatty acids and the potential mechanisms through which they may affect cardiovascular disease risk factors. RECENT FINDINGS: A cardioprotective effect for omega3 fatty acids is supported by prospective studies demonstrating an inverse association between fish intake and coronary heart disease mortality. Data from secondary prevention trials support a reduction in ventricular fibrillation as a primary mechanism for the decreased incidence of myocardial infarction. Clinical trials and experimental studies have shown that omega3 fatty acids have many other potentially important antiatherogenic and antithrombotic effects. Omega-3 fatty acids lower blood pressure and heart rate, improve dyslipidaemia, reduce inflammation, and improve vascular and platelet function. These favourable effects have until recently been primarily attributed to the omega3 fatty acid eicosapentaenoic acid, which is present in large amounts in fish oil. Controlled studies in humans now demonstrate that docosahexaenoic acid, although often present in lower quantities, has equally important anti-arrhythmic, anti-thrombotic and anti atherogenic effects. SUMMARY: Available evidence strongly suggests that eicosapentaenoic acid and docosahexaenoic acid have differing haemodynamic and anti-atherogenic properties. The effects of the two fatty acids may also differ depending on the target population. PMID- 16477173 TI - Conjugated linoleic acid and human health: a critical evaluation of the evidence. AB - PURPOSE OF REVIEW: This review critically evaluates studies investigating the effects of conjugated linoleic acid on human health, including effects on body composition, blood lipids, liver metabolism, insulin sensitivity and immune function. It focuses mainly on human intervention studies, but includes some reference to animal and cellular studies which provide insight into potential molecular mechanisms of action of conjugated linoleic acid. RECENT FINDINGS: Human studies continue to report inconsistent effects of conjugated linoleic acid on human health. Some of these reports are based on overinterpretation of marginal effects of supplementation. Recent data suggest that the effects of the substance may be isomer dependent and that cis-9,trans-11 and trans-10,cis-12 conjugated linoleic acids have opposing effects on blood lipids and on metabolism in adipocytes and hepatic cells. SUMMARY: Claims that conjugated linoleic acid is beneficial for health remain as yet unconvincing. Human studies investigating the effects of conjugated linoleic acid supplements have tended to use mixtures of isomers and have been inconsistent. More recent studies have attempted to use relatively pure preparations of single isomers and these studies suggest that the effects of conjugated linoleic acid may be isomer-specific. These recent data suggest a relative detrimental effect of trans-10,cis-12 conjugated linoleic acid on blood lipids. There appears to be little effect of conjugated linoleic acid on immune function and the effects on insulin sensitivity remain unclear. PMID- 16477174 TI - Obesity and cardiovascular disease in developing countries: a growing problem and an economic threat. AB - PURPOSE OF REVIEW: This review examines the rise of risk factors for cardiovascular disease, especially obesity, in developing countries and the implications for both health and economics. RECENT FINDINGS: In the majority of developing countries fertility and infant and child mortality have fallen markedly, and life expectancies have increased. Rapid urbanization, falling food prices, and globalization of economies have contributed to an increase in risk factors for chronic disease. Recent work indicates that the prevalence of these risk factors, including obesity, is rising faster than the historical experience of the West. The transition is affecting women in particular, and increases in risk factors are more marked among lower incomes in growing economies than among the wealthy. Rather than the stereotypical problem of the rich, chronic disease is now a problem for the poor. SUMMARY: Significant research in this area of global health has only been undertaken in the last decade. Additional field research is needed in every dimension of the transition, both to document the problem itself and to determine its economic and societal impact and cost effective responses. Two critical factors are virtually absent from existing work and should be emphasized. First, the impact of rising risk factors for, and mortality from, cardiovascular disease in the work force may imply a growing threat to continued economic progress. Second, because risk factor reduction requires society-wide strategies, broad public-private coalitions will be needed to mobilize sectors beyond healthcare. PMID- 16477175 TI - Nutrition support in critically ill patients: switching to evidence based. PMID- 16477176 TI - Glycemic control and nutrition in the intensive care unit. AB - PURPOSE OF REVIEW: Hospital clinicians frequently encounter hyperglycemia due to diabetes or the stress of critical illness in patients who are receiving nutrition support. RECENT FINDINGS: A growing body of evidence suggests that hyperglycemia in the hospital is associated with adverse outcomes (e.g. disability after acute cardiovascular events, infection and death) and that improvement in outcomes can be achieved with improved glycemic control or insulin. Therefore, familiarity with the implications of hyperglycemia and with its treatment are essential for clinicians practicing in hospital settings. SUMMARY: Questions persist regarding the optimal glucose goal range in differing patient groups. In addition, while the technology to deliver glycemic control in intensive care unit settings is widely available, data are limited about effective and safe insulin infusions. Research should focus on the risks and benefits of providing nutrition support in this group of patients, optimal glucose goal ranges, and on methods of achieving desired glucose goal ranges. PMID- 16477177 TI - Molecular aspects of insulin therapy in critically ill patients. AB - PURPOSE OF REVIEW: This review provides an overview of molecular mechanisms involved in beneficial effects of insulin in insulin resistant critically ill patients. RECENT FINDINGS: Intense insulin therapy reduced morbidity in critically ill patients. Insulin acts by two major molecular pathways: reduction of the inflammation process induced by free fatty acid excess in tissues and decrease of reactive oxygen species production induced by hyperglycemia. By these actions, insulin preserves mitochondrial function, enhances adiponectin secretion and probably modulates AMP-activated protein kinase activity, which in turn depletes lipid depots in tissues and restores glucose uptake and oxidation. Furthermore, it was recently established that insulin prevents microcirculation alteration and subsequent cellular hypoxia by reducing inducible nitric oxide synthase expression and activity in the endothelium. So, insulin beneficial effects in critically ill patients are dependent on metabolic and non-metabolic molecular pathways. SUMMARY: Critically ill patients requiring intensive care for more than a few days have a high risk of death. A tight control of glucose levels by intense insulin therapy reduced morbidity in critically ill patients. Unraveling the molecular mechanisms of insulin will provide new insights into the pathogenesis of multiple organ failure and will allow novel therapeutic strategies to manage patients needing intensive care. PMID- 16477178 TI - Glucose, insulin and myocardial ischaemia. AB - PURPOSE OF REVIEW: The importance of glucose metabolism and insulin therapy during myocardial ischaemia is increasingly being investigated. Insulin is used to achieve a tight glucose control or as part of glucose-insulin-potassium therapy. We have reviewed (1) the physiological and physiopathological consequences of hyperglycaemia focusing on potential machanisms of myocardial ischaemia, (2) the effects of insulin on vascular tone, on the release of free fatty acids, on inflammatory pathways, on the switch of energy source and on apoptosis, and (3) clinical data reporting the effects of intensive insulin therapy and glucose-insulin-potassium solutions during myocardial ischaemia and ischaemic heart failure. RECENT FINDINGS: In addition to its known toxic cellular effects, hyperglycaemia increases the activity of inducible nitric oxide synthase and promotes inflammation. Conversely insulin exerts anti-inflammatory and anti apoptotic effects. Glucose-insulin-potassium solutions could improve survival after acute myocardial infarction or after surgery, according to recent meta analyses, but confirmation of these data is eagerly awaited. SUMMARY: Hyperglycaemia is toxic, while insulin is beneficial during acute myocardial ischaemia. Some recent evidence confirms a substantial benefit of insulin administered either alone to achieve a tight glucose control or as a component of glucose-insulin-potassium therapy. Further research is needed to confirm that tendency and to define the threshold of tight glucose control. PMID- 16477179 TI - Fish oil in the critically ill: from experimental to clinical data. AB - PURPOSE OF REVIEW: The aim of this paper is to describe recent relevant literature concerning the role of n-3 lipids derived from fish oil in clinical nutrition in an intensive care setting. RECENT FINDINGS: N-3 fatty acids compete with arachidonic acid for metabolism to lipid mediators and exert profound effects on second mediator generation and dependent cellular functions. In experimental models, dietary and parenteral use of fish oil was shown to protect the gut by increasing its perfusion. In contrast, use of immunonutrition including fish oil in critical ill patients or patients with severe sepsis may exert an excess mortality. Using parenteral fish oil in surgical patients promising data became available. In septic patients, immunologic effects of fish oil-based lipid emulsions have been found and intravenous supplementation with fish oil may have a beneficial impact on mortality and length of stay. For both patient groups, however, prospective data from randomized trials are lacking. SUMMARY: N-3 lipids exhibit strong immunologic properties. They offer the possibility to counterbalance the negative effects of conventional n-6 fatty acids. Recent studies exhibit positive effects of intravenous use of fish oil on immunologic functions and clinical parameters in surgical and septic patients. PMID- 16477180 TI - Ergonomic and economic aspects of total parenteral nutrition. AB - PURPOSE OF REVIEW: Ergonomics in total parenteral nutrition include the work performed in the (hospital) pharmacy and on the medical ward. This article reviews the developments in total parenteral nutrition ergonomics and the related cost-savings. RECENT FINDINGS: Research focuses on the ergonomic advantages of multi-compartment total parenteral nutrition bags compared with the multi-bottle system, of multi versus single-layered total parenteral nutrition bags and of the presence of a nutritional team and training in clinical nutrition to improve regimen prescription and delivery. SUMMARY: Three-compartment bags are safe, economic and ergonomic. It is important, however, to keep the knowledge of pharmacies to compound total parenteral nutrition for children and (adult) patients with specific pathologies. Research is ongoing in the development of bags with more than three compartments, to include for instance vitamins. This necessitates improvements in bag materials and wrapping. Progress can be made regarding total parenteral nutrition prescription and delivery, as well as in the incidence of related infections by promoting training in clinical nutrition and the implementation of a multidisciplinary nutritional support team. PMID- 16477182 TI - Bibliography. Current world literature. Lipid metabolism. PMID- 16477181 TI - Enteral access for nutrition in the intensive care unit. AB - PURPOSE OF REVIEW: Enteral nutrition is the preferred route for nutrition support in the intensive care unit setting. This is usually delivered through nasoenteric feeding tubes in patients with an otherwise functional gastrointestinal tract. Placement of nasoenteric feeding tubes, however, may be difficult in this setting. Nasoenteric feeding tubes may be placed by multiple methods, each with their particular advantages and disadvantages. This review summarizes the recent literature on different methods of nasoenteric feeding tube placement with emphasis on critically ill patients. RECENT FINDINGS: Bedside assisted methods using electromyogram, electrocardiogram, and magnetic fields to provide immediate positional feedback to help guide tube advancement appear promising. Bedside methods using specific protocols, modified feeding tubes, prokinetics or magnetic assistance were also successfully reported. None of these methods has been prospectively compared with more commonly practiced methods in large studies. Endoscopic nasoenteric tube placement methods including transnasal approaches using ultra-thin endoscopes have been recently described and appear to be equivalent to fluoroscopic placement. All these recently reported techniques, however, may require more specialized equipment or training than is currently widely available. SUMMARY: Feeding tubes can be placed using bedside, fluoroscopic, and endoscopic means. Novel bedside methods have been recently described using immediate positional feedback or new assistive methods. Endoscopic techniques have similar success rates to fluoroscopic techniques and provide data on upper gastrointestinal abnormalities. There is no clear universal standard method. When feeding tube placement is required the technique used depends on local institutional resources and expertise. PMID- 16477184 TI - Dissociative symptoms and trauma exposure: specificity, affect dysregulation, and posttraumatic stress. AB - Although dissociation is often described as a posttraumatic response, the actual statistical association between trauma exposure and dissociative symptoms is surprisingly small. This suggests that that some dissociative phenomena may be unrelated to trauma, or may be related in more complex ways. Analysis of the normative data for the Multiscale Dissociation Inventory revealed significant dissociative symptoms in only 8% of trauma-exposed individuals from the general population. However, 90% of those with at least one clinically significant dissociation scale on the Multiscale Dissociation Inventory reported a trauma history, and significant dissociation was found in only 2% of nontraumatized individuals. A history of interpersonal violence, number of different types of trauma exposure, posttraumatic stress, and affect dysregulation were univariate predictors of dissociative symptomatology in trauma-exposed participants, but only posttraumatic stress and affect dysregulation were multivariate predictors. Trauma is probably an important, but insufficient, condition for the development of dissociative symptomatology. Additional risk factors, such as high posttraumatic stress and/or reduced affect regulation capacities, may determine whether trauma exposure results in clinically significant dissociation. PMID- 16477183 TI - Affect regulation in borderline personality disorder. AB - Although difficulty with affect regulation is generally considered a core component of borderline personality disorder (BPD), surprisingly little research has focused on the nature of affect regulation and dysregulation in BPD. A random national sample of 117 experienced clinicians provided data on a randomly selected patient with BPD (N = 90) or dysthymic disorder (DD; N = 27). Clinicians described their patients using the Affect Regulation and Experience Q-sort Questionnaire Version, a psychometric instrument designed for expert informants to assess affect and affect regulation. BPD and DD patients appear to differ in both the emotions they experience and the ways they regulate or fail to regulate them. Whereas DD patients are characterized by negative affect, BPD patients are characterized by both negative affect and affect dysregulation, which appear to be distinct constructs. BPD patients also show distinct patterns of affect regulation, and subtypes of BPD patients show distinct affect regulation profiles of potential relevance to treatment. PMID- 16477185 TI - P300 is attenuated during dissociative episodes. AB - The present study examined the pathophysiology of dissociative phenomena using the P300 component of event-related potentials, quantitative electroencephalography (QEEG), and morphology measures of computed tomography scan. Event-related potentials during an auditory oddball paradigm and QEEG in resting state were recorded. Patients exhibited attenuation of P300 amplitudes compared with controls during dissociative episodes, but exhibited recovery to control levels in remission. Patients had a larger Sylvian fissure-brain ratio than did controls. QEEG findings revealed no significant differences between the patients and controls or between episodes and remission in the patient group. Attenuation of the P300 can be interpreted as the result of a negative feedback loop from the medial temporal lobe to the cortex, which decreases the amount of information flow, allocation of attentional resources, and updating of working memory to avoid both excessive long-term memory system activity in medial temporal lobe and resurgence of affect-laden memories. PMID- 16477187 TI - Psychometric properties of the defense style questionnaire (DSQ-40) in adolescents. AB - This study examined the psychometric properties of the Defense Style Questionnaire (DSQ-40) in adolescents. Internal consistency, factor structure, and discriminant and concurrent validity of the DSQ-40 were studied in 211 adolescent psychiatric outpatients aged 13 to 19 years and 199 age-matched and sex-matched controls. Principal components analysis yielded four internally consistent components: mature, neurotic, image-distorting, and immature defense styles. The outpatients reported more immature, image-distorting, and neurotic styles and less mature style than did the controls, suggesting adequate discriminant validity. As a demonstration of convergent and concurrent validity, the severity of psychiatric symptoms assessed by the General Health Questionnaire and psychosocial adjustment assessed by the Global Assessment of Functioning Scale correlated theoretically meaningfully with the different defense styles. The DSQ-40 appears to be a reliable and valid instrument for adolescents. PMID- 16477186 TI - Characteristics and predictors of full and partial recovery from generalized anxiety disorder in primary care patients. AB - The current study examined the naturalistic course of generalized anxiety disorder (GAD) in a sample of 113 primary care patients across a 2-year period. Initial diagnoses were established using structured clinical interviews according to DSM-IV diagnostic criteria. Results indicated that the majority of patients meeting DSM-IV diagnostic criteria for GAD were still symptomatic to some degree after 2 years of follow-up. Rates of full and partial recovery from GAD, however, were found to be higher than those reported for previous studies of GAD in psychiatric patients. Diagnostic comorbidity, severity of psychosocial impairment, and gender were found to be significantly associated with achieving full or partial recovery from GAD. Psychiatric treatment was not found to be associated with time to full or partial recovery from GAD symptoms, likely due to a treatment-biasing effect. These results underscore that GAD is a chronic and persistent illness in primary care patients. PMID- 16477188 TI - Depressive symptoms and associated psychopathology in urban adolescents: a cross cultural study of three countries. AB - The study aimed to compare cross-cultural trends of comorbid internalizing and externalizing psychopathology, prosocial beliefs, and perceptions of risk in adolescents with and without clinical levels of self-reported depressive symptoms. The Social and Health Assessment, a self-report survey, was conducted in a representative sample of 3309 adolescents 14 to 17 years old from urban communities in the United States (N = 1343), Belgium (N = 946), and Russia (N = 1009). In all three countries, girls reported higher levels of depressive symptoms than boys. The findings also demonstrate that in both genders, depressive symptoms were associated with increased levels of internalizing and externalizing problems, as well as lower levels of prosocial beliefs and low perceptions of harm from risk-taking behavior. Depressed boys had relatively higher levels of externalizing problems than depressed girls. Greater levels of internalizing problems observed in depressed youth, as compared with their nondepressed counterparts, were not gender-specific. Current findings suggest that the relationships between depression and comorbid psychopathology are not culture-specific and have similar patterns in different populations. PMID- 16477189 TI - Do people turn to religion in times of stress?: an examination of change in religiousness among elderly, medically ill patients. AB - This study examined the effect of health-related stress on changes in religiousness in a sample of elderly, medically ill patients. Patients admitted to Duke University Medical Center (N = 745) were interviewed at baseline and 3 month follow-up. Increases in illness severity (from baseline to follow-up) were associated with decreases in both organizational and private religiousness at follow-up. Effect of illness severity on organizational religiousness was statistically mediated by changes in physical activity, while its effect on private religiousness remained significant after controlling for physical activity. These findings encourage further research investigating causal relationships between stress and religion, as well as identifying measures of religiousness that may capture this construct in the medically ill population. PMID- 16477190 TI - An exploration of associations between separation anxiety in childhood and complicated grief in later life. AB - Recent studies have suggested that the vulnerability to complicated grief (CG) may be rooted in insecure attachment styles developed in childhood. The aim of this study was to examine the etiologic relevance of childhood separation anxiety (CSA) to the onset of CG relative to major depressive disorder, posttraumatic stress disorder, and generalized anxiety disorder in bereaved individuals. The Structured Clinical Interview for the DSM-IV, Inventory of Complicated Grief Revised, and CSA items from the Panic Agoraphobic Spectrum Questionnaire were administered to 283 recently bereaved community-dwelling residents at an average of 10.6 months postloss. CSA was significantly associated with CG (OR = 3.2; 95% CI, 1.2-8.9), adjusting for sex, level of education, kinship relationship to the deceased, prior history of psychiatric disorder, and history of childhood abuse. CSA was not significantly associated with major depressive disorder, posttraumatic stress disorder, or generalized anxiety disorder. PMID- 16477191 TI - Investigation of insight formation using narrative analyses of people with schizophrenia in remission. AB - Poor insight in schizophrenic patients is a common and multidimensional phenomenon. The purpose of this study was to explore the process of the formation of insight in patients with schizophrenia in remission. A qualitative design was used to analyze first-person narratives of 50 people with schizophrenia in remission. Face to face, semistructured interviews were conducted by investigators, and the narratives were transcribed verbatim. The results of our study showed that there were four stages of illness understanding in patients with schizophrenia: 1) the feeling that symptoms were unbearable or a loss of control, 2) comparisons of experiences with references, 3) perception that medication was working through trial and error experiments, and 4) awareness of illness after medication relieved symptoms. Insight formation is an active process in which schizophrenic patients gain knowledge about their medication, connect the association between their medication and symptoms, and realize they need to deal with issues. PMID- 16477192 TI - Persistent physiological reactivity in a pilot study of partners of firefighters after a terrorist attack. AB - Twenty-four female partners of firefighters participating in recovery efforts associated with the 1995 terrorist bombing in Oklahoma City were assessed 43 to 44 months later. Disaster experiences, psychiatric diagnoses, posttraumatic stress symptoms, and autonomic reactivity in response to an interview about the bombing were examined. Most of the participants with postbombing disorders suffered from pre-existing conditions. The majority found the bombing a "terrible" or "shocking" experience. One participant met all DSM-III-R symptom group criteria for bombing-related posttraumatic stress disorder, and 40% met both B (intrusive re-experiencing) and D (hyperarousal) criteria. More than one half of the sample exhibited autonomic reactivity on at least one measurement. Those who met symptom group criterion D evidenced greater autonomic reactivity than those who did not, suggesting a link between self-reported posttraumatic stress disorder symptoms of arousal and biological manifestations. Thus, it may be important to assess partners of disaster recovery workers for mental health and physiological consequences related to their indirect exposure as these may persist years after the event, even in the absence of a diagnosable mental disorder. PMID- 16477193 TI - Personal mastery attenuates the effect of caregiving stress on psychiatric morbidity. AB - This study examined the protective effects of personal mastery on the relations between both objective and subjective stress and psychiatric morbidity in 79 spousal Alzheimer caregivers. Results indicated that with low mastery, the relations between patient problem behaviors and caregiver psychiatric symptoms was significant (t[71] = 2.03; p = 0.046). However, with high mastery, no significant association was found (t[71] = -0.76; p = 0.452). Similarly, the relations between role overload and psychiatric morbidity was significant when mastery was low (t[71] = 2.22; p = 0.029), but not high (t[71] = -1.49; p = 0.140). These results suggest that caregivers with a greater sense of personal mastery may be protected from the negative effects of caregiver stress. PMID- 16477194 TI - Childhood trauma and dissociation in female patients with schizophrenia spectrum disorders: an exploratory study. AB - The few studies that have investigated the relationship between trauma and dissociative symptoms in patients with schizophrenia have not assessed the role of the severity of psychotic symptoms. The current study examined correlations among five domains of childhood trauma and dissociative symptoms in 30 female patients with schizophrenia spectrum disorders, using the Dissociative Experiences Scale and the Childhood Trauma Questionnaire. Psychotic symptoms were measured by the Positive and Negative Syndrome Scale. Consistent with previous studies, high levels of childhood traumatic experiences were found (Childhood Trauma Questionnaire total score M = 48.5, SD = 18.3). Physical neglect and emotional abuse showed significant correlations with dissociative symptoms at admission. When patients were stabilized, about a month after admission, emotional abuse still showed a significant correlation with dissociative symptoms. However, in contrast to previous findings, Dissociative Experiences Scale findings were not stable over time. Our results confirm the relevance of childhood trauma in schizophrenic patients but also demonstrate the need to develop appropriate methodologies for measuring dissociation in this population. PMID- 16477195 TI - The Stigma Receptivity Scale and its association with mental health service use among bereaved older adults. AB - The purpose of this article was to determine whether the Stigma Receptivity Scale (SRS) predicts use of mental health services among community-dwelling bereaved older adults. We analyzed interviews of 135 people in Connecticut to evaluate whether three subscales and 12 SRS items were associated with access to any mental health service in the past 60 days using logistic regression analysis. Two SRS items predicted recent use of mental health services among bereaved individuals with and without complicated grief: receptivity to a bereavement support group (adjusted OR = 5.14; 95% CI, 1.11, 23.85) and individuals who were not concerned about meeting criteria for a mental illness (adjusted OR = 0.07; 95% CI, 0.01, 0.58). The SRS significantly predicted recent access to mental health treatment among bereaved elderly people. This type of measure could be used to determine those most likely in need of education and support to increase their likelihood of accessing mental health services. PMID- 16477197 TI - Shepherding change: how the market, healthcare providers, and public policy can deliver quality care for the 21st century. AB - BACKGROUND: Data are scarce that inform the ways consumers of health care and caregivers help improve the care that is delivered. The healthcare system is quite broad. To improve it, we first must understand it, understand its various subsystems, and understand how they shape individual behavior. DISCUSSION: Both consumers and providers can effectively improve health care. An example of an influential consumer is provided, focusing a successful effort to improve care in the intensive care unit. The overall model for improving outcomes assumes providers can be classified into high-quality and low-quality providers. The model then aims to increase the number of people exposed to high-quality caregivers. There are three primary levers for driving this change: using market forces, provider improvements, and policy change. This article touches briefly on the first and focuses on the second and third. CONCLUSION: A number of grassroots programs highlight ways providers can improve care by approaching patient safety and quality as a science. In addition, the Josie King Act and its legislative progeny provide some hope that a new policy environment can reward and reinforce providers' efforts to drive up safety and quality. PMID- 16477196 TI - Mixed lateral preference in posttraumatic stress disorder. AB - Recent research indicates that adults with posttraumatic stress disorder (PTSD) have a higher incidence of mixed laterality with respect to handedness than the rest of the population. To test if this relationship also occurs early in life, we evaluated children with history of interpersonal trauma. Fifty-nine traumatized children were evaluated with the Clinician Administered PTSD Scale for Children and Adolescents and the Edinburgh Handedness Inventory. Forty matched healthy controls were used for comparison. Increased mixed laterality was found in all children exhibiting symptoms of PTSD when compared with healthy controls, and children who met DSM-IV diagnostic criteria for PTSD had more mixed laterality than the subthreshold traumatized group (F = 7.71; df = 2,96; p = 0.001). Within the entire traumatized group, there was a positive correlation between PTSD symptom severity and mixed laterality. Mixed laterality was positively associated with PTSD symptoms in traumatized children, suggesting that neurological abnormalities may be related to degree of PTSD symptom expression. PMID- 16477198 TI - Team model: advocating for the optimal method of care delivery in the intensive care unit. AB - OBJECTIVE: To review published data on the team model of intensive care unit (ICU) care delivery. DESIGN: Nonexhaustive, selective literature search. SETTING: Review of literature published in the English language. PATIENTS/SUBJECTS: Humans cared for in ICUs. INTERVENTIONS: None. RESULTS: The team model for delivery of ICU care reduces mortality, ICU length of stay, hospital length of stay, and cost of care. Convincing data suggest that merely having daily rounds led by an intensivist enhances patient care significantly. Further improvements can be obtained by maintaining a nurse-to-patient ratio of no greater than 1:2, adding critical care pharmacists, and providing dedicated respiratory therapists to the ICU team. CONCLUSION: Current and looming shortages of all ICU healthcare providers is a barrier to universal implementation of the team model. Advocating for the ICU team model for critical care delivery requires local, regional, national, and international activities for success. PMID- 16477199 TI - Intensive care unit physician staffing: financial modeling of the Leapfrog standard. AB - OBJECTIVE: To evaluate from a hospital's perspective the costs and savings, over a 1-yr period, of implementing The Leapfrog Group's Intensive Care Unit Physician Staffing (IPS) standard compared with the existing standard of nonintensivist staffing in adult intensive care units. DESIGN: Using published data, we developed a financial model of costs and savings for 6-, 12- and 18-bed intensive care units using conservative estimates for all variables. Sensitivity analyses, including a best-case and worst-case scenario, were performed to evaluate the impact of changing assumptions on the outcome of the model. SETTING: Nonrural hospitals in the United States. PATIENTS: All adult intensive care unit patients. INTERVENTIONS: The IPS standard requires that intensive care units have a dedicated intensivist present during daytime hours. Outside of these hours, an intensivist must be immediately available by pager, and a physician or "physician extender" must be in the hospital and able to immediately reach intensive care unit patients. MEASUREMENTS AND MAIN RESULTS: Cost savings ranged from $510,000 to $3.3 million for 6- to 18-bed intensive care units. The best-case scenario demonstrated savings of $4.2-13 million. Under the worst-case scenario, there was a net cost of $890,000 to $1.3 million. CONCLUSIONS: Financial modeling of implementation of the IPS standard using conservative assumptions demonstrated cost savings to hospitals. Only under worst-case scenario assumptions did intensivist staffing result in additional cost to hospitals. These economic findings must be interpreted in the context of significant reductions in patient morbidity and mortality rates also associated with intensivist staffing. Given the magnitude of its clinical and financial impact, hospital leaders should be asking "how to" rather than "whether to" implement The Leapfrog Group's ICU Physician Staffing standard. PMID- 16477200 TI - Critical care nursing: workforce issues and potential solutions. AB - OBJECTIVE: To review the history and development of critical care nurses, the supply and demand issues related to critical care nursing, critical care nursing's contribution to patient outcomes, and recommendations to ensure a steady and strong workforce. DATA SOURCE AND SELECTION: Information presented in this article is based on a review of past and current literature including international and U.S. government reports, professional publications, monographs, newspapers, and journal articles identified by MEDLINE and CINAHL databases. The Internet was used to contact international and national professional organizations and specialists. DATA EXTRACTION AND SYNTHESIS: Original and selected review articles and guideline documents were reviewed for references to critical care nurses and their role on the multiprofessional critical care team. CONCLUSIONS: Critical care nurses are an essential and vital aspect of the critical care team. Nurses contribute to improved patient outcomes, reduced morbidity and mortality, reduced complications and errors, and reduced overall costs. More than 400,000 nurses practice in critical care, and additional opportunities exist and will develop. The challenge is to ensure an adequate supply of appropriately trained staff. PMID- 16477201 TI - Respiratory care manpower issues. AB - OBJECTIVE: Although respiratory care is a relatively new profession, its practitioners are deeply involved in providing patient care in the critical care. In preparation for writing this article, we sought to explore the respiratory therapy manpower needs and activities designed to fulfill those needs in critical care practice. MATERIALS AND METHODS: We began by delineating the historical development of respiratory care as a profession, the development of its education, and the professional credentialing system. We then conducted several literature reviews with few articles generated. We requested and received data from the American Association for Respiratory Care (AARC), The National Board for Respiratory Care (NBRC), and the Committee on Accreditation of Respiratory Care education (CoARC) relative to their membership, number of credentialed individuals, and educational program student and graduate data for 2000 through 2004. We then conducted two electronic surveys. Survey 1 was a six-item survey that examined the use of mandatory overtime in respiratory care departments. We used a convenience sample of 30 hospitals stratified by size (or=500 beds). Survey 2 was a five-item instrument distributed by blast E-mail to the Society of Critical Care Medicine's Respiratory Care Section members and members of the RC_World list serve. This survey elicited 51 usable and non-duplicative responses from geographically and size-varied institutions. We analyzed these data in several ways from distribution analysis to one-way analysis of variance procedure and appropriate post hoc analysis techniques. Where appropriate, a matched-pairs analysis was performed and these were compared across the variables intensive care unit (ICU) beds per actual number of respiratory care practitioners (RCPs) and ICU beds per preferred number of RCPs. RESULTS: The data gathered from the professional organizations indicated a relatively stable attrition rate (35.2%+/-1.7-3.1%), even in the face of varying enrollments (6,231 in 2004 vs. 4,589 in 2002). In survey 1, we looked at the institution of mandatory overtime policies and their use in 30 size-stratified hospitals. Mandatory overtime was selected as a survey topic under the supposition that manpower shortages might lead to the development of such procedures and also to their utilization. Fourteen of the 30 hospitals responding indicated that they had a policy addressing mandatory over time. Of the 14 hospitals with policies, only ten had disciplinary actions specific to refusing the overtime. Seven of the 30 hospitals indicated that they used mandatory overtime monthly of more frequently. Survey 2 data revealed that there was a wide variation in bed size, number of ICUs, and number of RCP staff assigned to the ICU. Serendipitously, our 51 responding centers were distributed among small (16), medium (19), and large (16) hospitals in a manner that appeared to reflect the national distribution pattern. We were able to use these data to develop a closeness of fit diagram ICU beds to preferred numbers of RCPs (DF=48; p<.0001; RSq=0.77; RMSE=4.114). The number of beds per preferred number of RCPs was 9.445 to 1.0 while the actual bed to RCP ratio was 10.75 to 1. CONCLUSION: This article provides a short history of the development of respiratory care and its historical relationship with critical care. We have, perhaps for the first time, provided a unified data set of key demographic information from the three professional bodies guiding the development of the respiratory therapy profession. This data set provides time-linked data on admissions and graduations from the CoARC, membership numbers for the AARC, and the numbers of active credentialed RCP from the NBRC. By two focused surveys, we were able to show that while mandatory overtime is a common practice in respiratory care departments, it was not overwhelming utilized. We also learned that in most hospitals, regardless of bed size, there is a perceived need for 1.3 RCPs more than the actual staff and that it appears that the critical staffing level between actual to preferred RCP to beds is between 9 and 11 beds. PMID- 16477202 TI - The critical care clinical pharmacist: evolution of an essential team member. AB - OBJECTIVE: To review the history, training requirements, contributions to patient care outcomes, and workforce issues of critical care pharmacists. DATA SOURCE AND SELECTION: Literature obtained through Medline search with manual cross referencing. DATA EXTRACTION AND SYNTHESIS: Original and selected review articles and guideline documents were reviewed for references to critical care pharmacists and their role on the multiprofessional critical care team. CONCLUSIONS: Critical care pharmacists are recognized as essential members of the critical care team as a result of contributions to medication safety, improved patient outcomes, and reduced drug costs and as a source of drug information and provider of education. A growing number of pharmacists practice in critical care. Additional opportunities exist and can be met if an adequate supply of trained specialists can be developed. PMID- 16477203 TI - Critical care and disaster management. AB - BACKGROUND: In recent years, there has been a great deal of attention paid to preparing the healthcare system to handle disasters, in particular terrorist events. Most of the attention has focused on the first responders and the initial emergency management. Depending on the nature of the disaster, however, large numbers of patients may be critically ill. DISCUSSION: In a contagious event, there may be a continuous stream of new patients requiring critical care support, overwhelming our current intensive care unit capacity. Planning needs to start now to develop processes that will enable us to expand our intensive care unit capacity, and likely adapt our standard of care, in the event that a natural or man-made disaster results in two, three, or more times the number of critically ill patients than our system can currently handle. Using the processes and resources we are currently using to improve patient safety can provide a framework for developing the necessary processes. CONCLUSION: The Society of Critical Care Medicine (SCCM) can provide valuable expertise and educational programs to facilitate the needed disaster management planning. PMID- 16477204 TI - Providing critical care during a disaster: the interface between disaster response agencies and hospitals. AB - OBJECTIVE: Recent natural disasters have highlighted shortfall areas in current hospital disaster preparedness. These include the following: 1) insufficient coordination between hospitals and civil/governmental response agencies; 2) insufficient on-site critical care capability; 3) a lack of "portability" of acute care processes (i.e., patient transport and/or bringing care to the patient); 4) education shortfalls; and 5) the inability of hospitals to align disaster medical requirements with other competing priorities. CONCLUSIONS: Definition of the roles and responsibilities of a hospital during a disaster requires additional planning precision beyond the prehospital response phase. Planners must also better define plans for circumstances when or if a hospital is rendered unusable. Disaster medical training of hospital personnel has been inadequate. This article details the specifics of these issues and outlines various potential approaches to begin addressing and formulating remedies to these shortfalls. PMID- 16477205 TI - Proceedings adapted from a payor panel. AB - BACKGROUND: This panel featured four representatives from the healthcare industry and government, offering an opportunity for critical care professionals to pose questions and discuss issues and concerns relevant to anyone caring for critically ill and injured patients today. A brief biography is provided for each panelist. DISCUSSION: The Society of Critical Care Medicine Advocacy Committee recognized that there are not enough opportunities for clinicians and other members of the critical care team to discuss questions or issues with their counterparts on the payor side of providing clinical care. That is, much of the difficulty faced by providers after providing critical care services could be resolved if the channels of communication were opened, and so a payor panel was organized to start the process. CONCLUSION: Each of the panelists provided a prepared statement on issues relevant to critical care, as evident from their respective roles. Specific scenarios and other suggestions regarding payment policy, coding, and quality of care are provided. PMID- 16477206 TI - Critical care workforce: a policy perspective. AB - BACKGROUND: Workforce studies may be disconnected from the policies that affect supply and demand for health professionals. Sporadic review of the physician workforce in the last century has led to wide swings in perception about its adequacy. However, workforce research has influenced federal policy as well as the policies of institutions responsible for training and regulation of physicians. This discussion is intended to address workforce issues in the context of public policy at the federal level. It is also intended to serve as a starting point for new approaches to shaping workforce policy. DISCUSSION: The supply of and demand for physicians and other health professionals are affected by a number of factors that may or may not be under the control of policymakers and health professionals themselves. Productivity, practice patterns, the aging of the workforce and patients, and other major determinants are only minimally affected by most government policy. Despite several attempts throughout the 1980s and 1990s, demand for health care has been particularly difficult to control for policymakers. In contrast to the United States, most developed nations are extensively involved in the planning of the healthcare workforce, including the specialty mix, the number of physicians, and the number of other health professionals. CONCLUSION: There are many barriers to successful workforce policy. Successful public policy change often involves multiple stakeholders, in and out of government. The task before those concerned about workforce issues is to educate policymakers about how changes in the physician workforce will affect cost, access, and quality, and to impress upon them that serious efforts to improve quality of care and reduce costs will not be effective unless qualified physicians are there to provide that care. PMID- 16477207 TI - Evaluation and management codes: from current procedural terminology through relative update commission to Center for Medicare and Medicaid Services. AB - BACKGROUND: Physicians should have a working knowledge of the process by which patient care codes are created and subsequently assigned values. The Society of Critical Care Medicine has representatives on the national committees that focus on code creation and definition and on assignment of relative value units. In addition, a better understanding of documentation requirements and the audit process will facilitate improved compliance and minimize liability. DISCUSSION: The authors discuss the current procedural terminology (CPT) process for defining care codes and the relative update commission (RUC) process for assigning values to those codes, with each code assigned a separate value in three separate categories. Steps for managing any concern or dispute about billing, denials, or an audit are subsequently addressed. Tenets of proper documentation are discussed, and some future developments are identified that are likely to affect critical care. CONCLUSION: Knowledge of the procedures by which care codes are defined and valued is necessary for using these codes properly, as well as for addressing needs unmet by existing codes. Preventing audits is the best approach to proper coding and billing, and documentation is key. PMID- 16477208 TI - Unsustainable growth rate: physician perspective. AB - OBJECTIVE: To describe the sustainable growth rate history and calculations and why the present approach is not sustainable. BACKGROUND: The sustainable growth rate was created to help control the growth rate in healthcare expenditures. Presently, it is responsible, at least in part, for reductions in physician payment. Two components of the sustainable growth rate make it unsustainable in its present form. These components are 1) the inability to go back and correct for values based on the wrong assumptions and 2) the inclusion of drug-related costs into the sustainable growth rate. DISCUSSION: Few physicians have a full grasp of how their payment is structured from a mechanistic standpoint. A significant component of determining physician payment is established by the relative value unit, and how these are derived has been discussed in another article in this supplement. Once the relative value unit has been established, it is multiplied by the established conversion factor. A major component of the conversion factor is the sustainable growth rate. This article attempts to explain how the sustainable growth rate came into being, how it is calculated, and how it is flawed and contributing to decreasing physician payments. A few possible remedies are discussed, and the potential ramifications of those remedies on the physician community and the premiums of Medicare patients are examined. PMID- 16477209 TI - Economics of critical care: Medicare part A versus part B payments. AB - OBJECTIVE: To review the effect of Medicare part A payments (to hospitals) and part B payments (to providers) on critical care in the United States. DATA SOURCE AND SELECTION: Sources included U.S. government data and published literature reviewing the impact of Medicate payments on critical care. DATA EXTRACTION AND SYNTHESIS: Government data were reviewed to assess the history and status of reimbursement to hospitals and healthcare providers. These data, along with input from published literature, was used to assess the adequacy of current and projected Medicare reimbursements and the implications of these payments. CONCLUSION: Medicare payments to hospitals, particularly for critically ill patients, seem to fall short of the costs of caring for these patients. Reimbursements to providers seem more encouraging, although the opportunity exists to improve in this area as well. PMID- 16477210 TI - The future of hospital economic health. AB - OBJECTIVE: To evaluate factors which may influence the economic future of academic medical centers (AMCs). DATA SOURCE AND SELECTION: A literature search was performed to identify publications which reviewed the areas of revenue sources for AMCs, costs and expenses incurred by these institutions, and mechanisms for optimizing institutional economic stability. DATA EXTRACTION AND SYNTHESIS: Data were reviewed and evaluated in two primary contexts: hospital revenues and organizational and administrative factors influencing hospital economic health. CONCLUSIONS: Increasing economic stress will require AMCs to make efforts both to increase revenue through a variety of mechanisms and to minimize expenses without compromising their mission or impairing worker morale. PMID- 16477211 TI - B-cell tolerance following ABO-incompatible infant heart transplantation. AB - Infant heart transplantation is highly successful therapy for severe congenital heart malformations and cardiomyopathies. However the donor organ shortage is particularly crucial for this age group. We have demonstrated that some aspects of human immaturity can be exploited for infant organ transplantation, allowing safe transplantation of organs that would be "incompatible" in mature individuals, and as a window of continued susceptibility to induction of immunologic tolerance. PMID- 16477212 TI - B-cell tolerance. AB - Autoreactive B cells are actively tolerized to more abundant self-antigens by a series of checkpoints involving receptor editing, deletion, anergy and competition for growth factors. In contrast, B cells reactive against rare, sequestered or tissue specific self-antigens remain functionally naive. During an immune response, the autoimmune danger from these cells is countered by a variety of mechanisms comprising control of self-antigen presentation, limitation of immunogenic and tolerogenic costimuli including T cell help, homeostatic control of growth and strict regulation of germinal centre reactions. In this overview we consider how knowledge of these checkpoints may be used to gain a better understanding of transplant tolerance and the generation of alloantibodies. PMID- 16477213 TI - Regulation of cholesterol homeostasis in solid organ transplantation. PMID- 16477214 TI - Systematic analysis of composite tissue allograft rejection. PMID- 16477215 TI - Combination of voriconazole and caspofungin as primary therapy for invasive aspergillosis in solid organ transplant recipients: a prospective, multicenter, observational study. AB - BACKGROUND: : The efficacy of the combination of voriconazole and caspofungin when used as primary therapy for invasive aspergillosis in organ transplant recipients has not been defined. METHODS: : Transplant recipients who received voriconazole and caspofungin (n=40) as primary therapy for invasive aspergillosis (proven or probable) in a prospective multicenter study between 2003 and 2005 were compared to a control group comprising a cohort of consecutive transplant recipients between 1999 and 2002 who had received a lipid formulation of AmB as primary therapy (n=47). In vitro antifungal testing of Aspergillus isolates to combination therapy was correlated with clinical outcome. RESULTS: : Survival at 90 days was 67.5% (27/40) in the cases, and 51% (24/47) in the control group (HR 0.58, 95% CI, 0.30-1.14, P=0.117). However, in transplant recipients with renal failure (adjusted HR 0.32, 95% CI: 0.12-0.85, P=0.022), and in those with A. fumigatus infection (adjusted HR 0.37, 95% CI: 0.16-0.84, P=0.019), combination therapy was independently associated with an improved 90-day survival in multivariate analysis. No correlation was found between in vitro antifungal interactions of the Aspergillus isolates to the combination of voriconazole and caspofungin and clinical outcome. CONCLUSIONS: : Combination of voriconazole and caspofungin might be considered preferable therapy for subsets of organ transplant recipients with invasive aspergillosis, such as those with renal failure or A. fumigatus infection. PMID- 16477216 TI - Cholesterol absorption and synthesis in pediatric kidney, liver, and heart transplant recipients. AB - BACKGROUND: Hypercholesterolemia after organ transplantation is common. Previously, we observed higher serum total cholesterol (TC) concentrations in our pediatric kidney than liver or heart transplant recipients. To find an explanation to the observed difference, our kidney recipients' cholesterol synthesis and absorption efficiency was compared to those of liver and heart recipients. METHODS: Serum noncholesterol sterol ratios (10 x mmol to the mol of TC, surrogate estimates of hepatic cholesterol synthesis and intestinal absorption) were studied in 50 pediatric kidney, 25 liver and 12 heart transplant recipients without diabetes or cholestasis, and in 29 controls. RESULTS: The kidney recipients had lower Delta-cholesterol (P=0.031), similar lathosterol and higher desmosterol ratios (markers of cholesterol synthesis) (P=0.020), and similar campesterol and sitosterol ratios (markers of cholesterol absorption) when compared to the controls. The liver recipients had lower campesterol ratios than the kidney recipients and controls (P=0.002). Glomerular filtration rates were not associated with the ratios of noncholesterol sterols. Multivariate analysis showed markers of cholesterol synthesis to be lower and absorption to be higher in the kidney than the liver or the heart transplant recipients. Weight adjusted dosages of immunosuppressive agents were associated with some ratios of noncholesterol sterols and cholestanol though these varied between the transplant recipient groups. CONCLUSIONS: Serum TC concentration in kidney recipients was not significantly associated with absorption efficiency or synthesis of cholesterol, though kidney transplantation was associated with low synthesis and high absorption efficiency of cholesterol. Immunosuppressive therapy with cyclosporine and methylprednisolone may modulate absorption efficiency and synthesis of cholesterol. PMID- 16477217 TI - Posttransplant diabetes mellitus in kidney transplant recipients receiving calcineurin or mTOR inhibitor drugs. AB - BACKGROUND: The aim of this study was to evaluate the incidence and risk factors for posttransplant diabetes mellitus (PTDM; defined as new insulin use and/or new hyperglycemia) in 528 kidney recipients using different immunosuppressive agents. METHODS: Maintenance therapy included mycophenolate mofetil or azathioprine plus glucocorticoids in combination with Group I cyclosporine (263); Group II tacrolimus (60); or Group III sirolimus (205). RESULTS: The mean follow-up was 39.2 (range 9.0-103.8) months. Overall, the number of patients needing insulin was 7.4% (39/528). The incidences for Groups I, II, and III of 7.6%, 11.7%, and 5.9%, respectively, were not statistically different. Characteristics of patients with PTDM included older age (P=0.007); greater body weight (kg) at transplant, 6 months, and 12 months, respectively (P<0.001); greater BMI (kg/m2) at transplant, 6 months, and 12 months, respectively (P<0.001); more acute rejection episodes 28.2% vs. 13.5% (P=0.012); and increased incidence in African Americans (P=0.03). Multivariable analysis demonstrated increased risk for PTDM (defined as new insulin use) for tacrolimus, (hazard ratio [HR] 3.794, P=0.007); treated rejections (HR 2.491, P=0.0115); age (HR 1.407, P=0.0116); and BMI (HR 1.153, P<0.0001). New insulin use occurred sooner and with less total glucocorticoid dose for tacrolimus patients. If PTDM is defined as all cases of new hyperglycemia, then no immunosuppressive drug group demonstrated an increased risk. CONCLUSION.: The risk for developing PTDM is greatest among older recipients, and those obese at the time of transplant; those given steroid pulse therapy were at exceptionally high-risk. PTDM risk reduction should focus on weight loss in the obese end-stage renal disease population prior to transplant. PMID- 16477218 TI - Differential expression of profibrotic and growth factors in chronic allograft nephropathy. AB - BACKGROUND: Chronic allograft nephropathy (CAN) is a multifactorial process, where both immunological and nonimmunological factors play roles. Microarrays detect thousands of genes simultaneously. METHODS: We have analyzed gene expression profiles of 16 kidney transplant biopsy samples with CAN by high density oligonucleotide microarrays, comparing to six normal transplant biopsies. Eight CAN biopsies showed nodular arteriolar hyalinization and one was positive for C4d staining. RESULTS: Hierarchical clustering analysis of the 22 biopsies revealed differential gene expression patterns in CAN versus the control biopsies. However, microarray analysis did not reveal differential gene expression patterns in patients with or without arteriolar hyalinization. Fifty percent of the 100 genes with highest hybridization intensities in a C4d positive sample were related to cellular and humoral immune response. Although 212 genes were upregulated a minimum of 1.5-fold, 112 genes were downregulated in CAN samples. There was differential expression of profibrotic and growth factors that while transforming growth factor-beta induced factor, thrombospondin 1, and platelet derived growth factor-C were up-regulated, vascular endothelial growth factor, epidermal growth factor, and fibroblast growth factors 1 and 9 were downregulated. Selected differentially expressed genes were confirmed in microdissected samples by real-time quantitative PCR. Immunopathologic examination of biopsies revealed strong TGF-beta but decreased glomerular VEGF expression in CAN. CONCLUSION: Microarrays might be an important tool to uncover the mechanisms of multifactorial diseases, such as CAN. PMID- 16477219 TI - Clinical outcomes of living donor liver transplantation for hepatitis C virus (HCV)-positive patients. AB - BACKGROUND: Whether hepatitis C virus recurrence occurs earlier and with greater severity for living donor liver transplantation (LDLT) than for deceased donor liver transplantation (DDLT) has recently become a subject of debate. METHODS: We retrospectively evaluated clinical outcomes for a cohort of 91 HCV-positive patients who underwent LDLT at Kyoto University with a median follow-up period of 25 months. RESULTS: Overall 5-year patient survival for HCV patients was similar to that for non-HCV patients (n=209) who underwent right-lobe LDLT at our institute (69% vs. 71%). Survival rate of patients without HCC (n=34) tended to be better than that of patients with HCC (n=57) (82% vs. 60%, P=0.069). According to annual liver biopsy, rate of fibrosis progression to stage 2 or more (representing significant fibrosis) was 39% at 2 years after LDLT. Univariate analysis showed that female recipient and male donor represented significant risk factors for significant fibrosis. Progression to severe recurrence (defined as the presence of liver cirrhosis (F4) in a liver biopsy and/or the development of clinical decompensation) was observed in five patients. CONCLUSIONS: Postoperative patient survival was similar for HCV-positive and -negative recipients in our adult LDLT series. Rates of progression to severe disease due to HCV recurrence seemed comparable between our LDLT recipients and DDLT recipients described in the literature. Although longer-term follow-up is required, our results suggest that LDLT can produce acceptable outcomes also for patients suffering from HCV-related cirrhosis. PMID- 16477220 TI - Conversion to sirolimus and mycophenolate can attenuate the progression of bronchiolitis obliterans syndrome and improves renal function after lung transplantation. AB - BACKGROUND: Bronchiolitis obliterans syndrome (BOS) is the major problem after lung and heart-lung transplantation (LTx/HLTx). Sirolimus (Sir) and Mycophenolate (MMF) showed a promising efficacy in the treatment of BOS in animal models. The first clinical experience in converting LTx/HLTx-recipients with BOS from calcineurin inhibitor-(CNI)-based immunosuppression to a Sir-MMF based immunosuppression is reported herein. METHODS: Six LTx- and five HLTx-recipients (eight men; 0.9 to 8 years after transplantation) with CNI-based immunosuppression (plus MMF) in whom BOS was diagnosed were included in the study. Mean patient age was 37+/-13 years (range 17-62 years). Sir was started with 6 mg and continued adjusted to according target trough levels (8-14 ng/ml). Subsequently, the CNIs were tapered down and finally stopped. Follow up included self determined pulmonary function tests, microbiological screening, chest radiographs, and laboratory studies RESULTS: Two acute rejection episodes occurred during the study period. The incidence of infection was 2.2+/-1.3 infections/patient-year after conversion. Mean FEV1 decreased after a mean follow up of 14.8+/-1.4 months: from 2.1+/-0.7 l prior conversion to 1.3+/-0.6l after conversion (P=0.03). However, graft function remained stable in three patients and progression of BOS slowed down in three patients. Overall, 2 of 10 patients died due to ongoing BOS while awaiting retransplantation CONCLUSIONS: After BOS was diagnosed, conversion to MMF and Sir stabilized graft function only in some of the converted patients. Therefore, earlier administration of Sir-based immunosuppression might be a more promising approach. Whether conversion to CNI free immunosuppression can actually ameliorate the extent or progression of BOS has to be investigated in randomized trials. PMID- 16477221 TI - Elevated HDL cholesterol is functionally ineffective in cardiac transplant recipients: evidence for impaired reverse cholesterol transport. AB - BACKGROUND: Cardiac transplant recipients frequently have high plasma HDL levels but it is unclear whether these promote a cardioprotective profile. METHODS: Parameters of reverse cholesterol transport and endothelial function were compared in 25 cardiac transplant recipients with low (<1.4 mmol/L; n=11) or high (>1.4 mmol/L; n=14) plasma levels of HDL and in a reference healthy group. RESULTS: Patients with high HDL had lower levels of triglyceride and prebeta1-HDL and a higher proportion of large HDL particles. When normalized to apoA-I content, non-ABCA1-dependent cholesterol efflux from RAW 264.7 macrophage cells to plasma from high HDL patients was 33% lower when compared to plasma from patients with low HDL, whereas ABCA1-dependent cholesterol efflux was not impaired. Forearm vascular responses to acetylcholine and sodium nitroprusside were not influenced by HDL levels in these patients. Compared to a reference healthy group (n=26), cardiac transplant recipients had higher levels of triglyceride, lower levels of prebeta1-HDL and LCAT, and lower activities of cholesteryl ester transfer protein and phospholipid transfer protein. CONCLUSIONS: Hyperalphalipoproteinaemia in cardiac transplant recipients is associated with the formation of partially dysfunctional HDL. We conclude that high levels of HDL may not confer cardioprotection in this group of patients. PMID- 16477222 TI - Pretransplant human herpesvirus 6 infection of patients with acute liver failure is a risk factor for posttransplant human herpesvirus 6 infection of the liver. AB - BACKGROUND: Acute liver failure (ALF) is a significant cause of liver transplantation. We have previously reported that human herpesvirus-6 (HHV-6) was found in most livers of patients with ALF of unknown origin ending up with liver transplantation. In this study, we investigated the posttransplant HHV-6 infection of the liver graft in these patients. METHODS: Thirty-two patients transplanted due to ALF were included in this retrospective study. Twelve of the 15 patients with unknown cause and four of 17 patients with a known cause of ALF had HHV-6 antigens in the explanted liver. Altogether, 18 patients had some pretransplant evidence of HHV-6. After transplantation, the patients were frequently monitored for the viruses, and biopsy histology was performed in every case of graft dysfunction. HHV-6 was demonstrated in liver tissue by immunohistochemistry. RESULTS: During the follow-up of 6 months, hepatic HHV-6 infection was demonstrated in 9 of the 18 patients, at a mean 19 days (6-38 days) after transplantation. All patients with posttransplant HHV-6 showed graft dysfunction. In biopsy histology, seven out of these nine patients demonstrated viral infection, one of them also having CMV antigens in the liver. None of those patients without evidence of pretransplant HHV-6 showed HHV-6 in the posttransplant biopsies. Posttransplant HHV-6 was not treated and the virus had no effect on 1-year patient or graft survivals. CONCLUSION: Pretransplant hepatic HHV-6 infection of patients with ALF is a risk factor for posttransplant HHV-6 infection and liver dysfunction, but has no effect on 1-year graft or patient survival. PMID- 16477223 TI - Optimal portal venous circulation for liver graft function after living-donor liver transplantation. AB - BACKGROUND: Previous studies have shown poor outcome after living-donor liver transplantation (LDLT) as a result of excessive portal venous pressure (PVP), excessive portal venous flow (PVF), or inadequate PVF. We investigated optimal portal venous circulation for liver graft function after LDLT in adult recipients retrospectively. METHODS: Between June 2003 and November 2004, 28 adult patients underwent LDLT in our institution. We modulated PVP under 20 mmHg in these 28 cases by performing a splenectomy (n=4) or splenorenal shunt (n=1). The PVF and PVP were measured at the end of the operation. Compliance was calculated by dividing PVF by PVP. RESULTS: PVF and compliance showed a significant inverse correlation with peak billirubin levels after LDLT (r = -0.63: r=-0.60, P<0.01), and with peak international normalized ratio after LDLT (r=-0.41: r=-0.51, P<0.05). Compliance was higher in right-lobe graft with middle hepatic vein cases (148+/-27 ml/min/mmHg), and lower in left-lobe graft cases (119+/-50 ml/min/mmHg). CONCLUSIONS: Liver graft function was better when PVF and graft compliance were higher and PVP was maintained under 20 mmHg. PMID- 16477224 TI - Utility of HbA1c in the detection of subclinical post renal transplant diabetes. AB - BACKGROUND: We hypothesized that the use of HbA1c testing would help identify postrenal transplant diabetes (PTDM). METHODS: In all, 199 adult kidney transplant recipients at least 3 months posttransplant without previous history of diabetes or elevated fasting blood sugar were studied. Medical history, a fasting blood glucose, calcineurin inhibitor blood level, and HbA1c were obtained. Primary outcome was the incidence of subjects with HbA1c > or =6.1%. The covariates were use of cyclosporine or tacrolimus, time posttransplant, body mass index (BMI) at transplant and change since transplant, current steroid dose, history of graft rejection, current fasting glucose, age, and race. Proportions were compared between HbA1c <6 and > or =6.1% using Fisher's exact test. Means were compared using Student's t test. Logistic regression was used to identify risk factors associated with elevated HbA1c. RESULTS: Twenty subjects (10.1%) had an elevated HbA1c. High normal fasting glucose (P=0.003) and African American race (P=0.08, marginally significant) were found to be associated with an elevated HbA1c. Subjects with normal and abnormal HbA1c levels were otherwise similar. There was no difference in HbA1c in tacrolimus versus cyclosporine treated subjects or in the percent of subjects with elevated HbA1c between these groups. CONCLUSIONS: HbA1c levels were found to be more a more sensitive test than fasting blood glucose levels in PTDM, with 10.1% of all patients and 19.4% of blacks found to have an elevated HbA1c. HbA1c testing should be considered as a screening test for PTDM, especially in African Americans. PMID- 16477225 TI - Bloodstream infections among heart transplant recipients. AB - BACKGROUND: Heart transplant (HT) recipients are prone to life-threatening infections, including bloodstream infection (BSI), but information on this topic is particularly scarce. METHODS: We studied 309 consecutive HT performed at our institution between 1988 and 2003. We assessed the characteristics of each episode of BSI, prophylaxis and immunosuppression used, and possible related factors. RESULTS: Sixty episodes of BSI occurred in 15.8% of all HT recipients. Rates of BSI/transplanted patient decreased progressively throughout the study period: 21.2%, 14.3%, and 7.5% in each 5-year period (P=0.03). BSI episodes occurred a median of 51 days after transplantation. The main BSI origins were: lower respiratory tract (23%), urinary tract (20%), and catheter-related-BSI (16%). Gram-negative organisms predominated (55.3%), followed by Gram-positive (44.6%). Mortality was 59.2%, with 12.2% directly attributable to BSI. Independent risk factors for BSI after HT were: hemodialysis (OR 6.5; 95% CI 3.2 13), prolonged intensive care unit stay (OR 3.6; 95% CI 1.6-8.1), and viral infection (OR 2.1; 95% CI 1.1-4). BSI was a risk factor for mortality (OR 1.8; 95% CI 1.2-2.8). CONCLUSION: BSIs have decreased in HT recipients, but still contribute to mortality, mainly if related to pneumonia or polymicrobial infections. Reduction of early postoperative complications and viral infections are amenable goals that may further reduce BSI in this population. PMID- 16477226 TI - Platelet-derived growth factor gene polymorphism in recurrent hepatitis C infection after liver transplantation. AB - BACKGROUND: Recurrent hepatitis C virus (HCV) infection is particularly aggressive in the post liver transplantation setting, with rapid progression of liver fibrosis. Platelet-derived growth factor (PDGF) is reportedly involved in the pathogenesis of liver fibrosis. The aim of this study was to evaluate the possible contribution of molecular variants of the PDGF-B gene to recurrent HCV infection after liver transplantation. METHODS: DNA was extracted from peripheral blood mononuclear cells of 40 patients who underwent liver transplantation for chronic HCV infection and genotyped for polymorphisms in PDGF-B at positions +1135 (A to C) and +286 (A to G). Intrahepatic PDGF-B expression was detected by immunohistochemistry and assessed semiquantitatively. Forty-seven healthy individuals served as controls. RESULTS: Recurrent HCV infection occurred in 34 patients (85%) after a median interval of 10.5 months (range 1.5-60.0). A statistically significant difference was observed in the distribution of the PDGF B gene polymorphism at position +1135, but not +286 between patients and controls (P=0.05). The A/A genotype occurred at a highly significantly increased rate in patients with recurrent HCV infection than in those without (64.7% vs. 16.67%, P=0.0001), and in patients with severe than in those with nonsevere recurrence (100% vs. 53.85%, P=0.05). The expression level of intrahepatic PDGF-B was found to be highly correlated with the fibrosis stage (P<0.0001). Further analysis yielded a highly statistically significant relationship between the PDGF-B gene polymorphism at position +1135 and clinical parameters of disease severity. CONCLUSIONS: PDGF-B gene polymorphism appears to be associated with severe recurrent HCV infection after liver transplantation. PDGF-B may play an essential role in the development and progression of hepatic fibrosis. These findings, if confirmed, may have important therapeutic implications. PMID- 16477227 TI - Contrasting CD25hiCD4+T cells/FOXP3 patterns in chronic rejection and operational drug-free tolerance. AB - BACKGROUND: Although immunosuppression withdrawal in kidney recipients usually leads to rejection, in some patients it does not, leading to a state of clinical operational tolerance. METHODS: We compared these highly contrasted situations by analyzing blood cell phenotype and transcriptional patterns in drug-free spontaneously tolerant kidney recipients, recipients with chronic rejection, recipients with stable graft function under standard or minimal immunosuppression and healthy individuals RESULTS: The blood cell phenotype of clinically tolerant patients did not differ from that of healthy individuals. In contrast, recipients with chronic rejection had significantly less CD25hiCD4+T cells and lower levels of FOXP3 transcripts compared with clinically tolerant recipients. Patients with chronic rejection also displayed CD25-CD4+T cells expressing NKG2D+CD94+ and CD57+CD27-CD28- cytotoxic-associated markers (P<0.05). CONCLUSION: These data show that whereas clinically tolerant recipients displayed normal levels of CD25hiCD4+T cells and FOXP3 transcripts, chronic rejection is associated with a decrease in CD25hiCD4+T cells and FOXP3 transcripts, suggesting that clinically "operational tolerance" may be due to a maintained phenomenon of natural tolerance that is lacking in patients with chronic rejection. PMID- 16477228 TI - First case of toxoplasmosis following small bowel transplantation and systematic review of tissue-invasive toxoplasmosis following noncardiac solid organ transplantation. AB - BACKGROUND: Toxoplasmosis prophylaxis is standard following heart and heart lung transplantation, when an increased risk of allograft transmitted Toxoplasma is well-recognized. In contrast, prophylaxis and routine serologic evaluation of donors and recipients for Toxoplasma in noncardiac solid organ transplantation (SOT) is not recommended. We report the first case of disseminated toxoplasmosis following small bowel transplantation, presumably transmitted via the transplanted intestine and systematically review reported cases of toxoplasmosis in noncardiac SOT recipients to determine if current guidelines should be reconsidered. METHODS: Systematic MEDLINE review was performed for tissue invasive toxoplasmosis in noncardiac SOT recipients and analysis of clinical features, serologic status, and treatment regimens with respect to mortality. RESULTS: Fifty-two cases of toxoplasmosis in noncardiac SOT recipients were identified. Eighty-six percent developed disease within 90 days of transplantation. Presentation was nonspecific and consisted of fever (77%), respiratory distress (29%), neurologic manifestations (29%), and bone marrow suppression (26%). Multivariate analyses demonstrated that localized disease (odds ratio [OR]=37.36, 95% CI 1.85-754.85), treatment received (OR=1.814, 95% CI 1.193-3.480) and donor and recipient serostatus (OR=1.39, 95% CI 1.068-1.815) were predictors of survival. High-risk recipients (donor seropositive/recipient seronegative) developed disease earlier (16 days vs. 31 days P=0.002) and were less likely to survive (OR=0.14, 95% CI 0.03-0.69) than standard-risk recipients. CONCLUSIONS: Toxoplasmosis is recognized following noncardiac SOT. Reduction of morbidity and mortality necessitates knowledge of donor and recipient Toxoplasma serostatus, prophylaxis, early diagnosis, and treatment. The findings support a reconsideration of pretransplantation evaluation and prophylaxis strategies in SOT recipients. PMID- 16477229 TI - Composite tissue allotransplantation: classification of clinical acute skin rejection. AB - BACKGROUND: Composite tissue allotransplantation (CTA) is a recently introduced option for limb replacement and reconstruction of other nonreconstructible tissue defects. As with recipients of other allotransplants, CTA recipients can experience rejection episodes that are presumed to be mediated by immune mechanisms similar to those affecting solid organ grafts. However, a systematic examination of this process has not been performed, and there are no standardized criteria for the description of severity or type of rejection. METHODS: We collected biopsies from human limb allografts and abdominal walls in various stages of rejection for histological and immunohistochemical analysis to formulate a CTA rejection scheme. Biopsies were ranked by severity and reproducibility of the system was tested using a second set of biopsies. Tissue slides were examined blindly by three pathologists and the nonparametric Kendall coefficient of concordance (W) was used to assess the amount of agreement among the pathologists in their classification grades. RESULTS: Rejection initially appeared as a perivascular infiltrate progressing to involve the dermis. Arteritis was observed only in the medium to large size arteries of the subcutis. Myositis was seen occasionally. Perineural involvement without frank neuritis was present in advanced rejection. The infiltrate was predominantly CD4+ in milder cases and CD8+ in advanced cases. HLA-DR was minimally expressed in keratinocytes even in severe rejection. Kendall's W was 0.9375 (p< or =0.0001). CONCLUSIONS: Based on this survey, we proposed an initial classification system for acute rejection in the skin of a CTA and demonstrated that this system is easily reproduced by independent pathologists. PMID- 16477230 TI - Characterisation of collagen VI within the islet-exocrine interface of the human pancreas: implications for clinical islet isolation? AB - BACKGROUND: To optimize the methods used for human islet isolation for transplantation, it is important to improve our understanding of the structure of the islet-exocrine interface. In this study, the composition of collagen subtypes in the interface have been characterized and quantified in human pancreas. METHODS: Human adult pancreases were retrieved from older (mean age 55.7+/-3.0 yrs) and young donors (mean age 21.8+/-3.2 yrs). Tissue from the body of each pancreas was examined by quantitative immunohistochemistry. Collagen within the islet-exocrine interface was identified by immunolabeling for collagen I, IV, V or VI and islets identified either morphologically or by immunolabeling for insulin. Collagen subtypes were quantified and data expressed as collagen area at the interface relative to the islet area. Statistical analysis was by ANOVA or Mann Whitney U test. RESULTS: In older pancreases, collagen IV, V and VI were present throughout the islet-exocrine interface, whereas collagen I was more variable. The mean peri-islet collagen VI proportion was significantly greater than that of collagen I or IV. Mean islet area and the proportional collagen VI content in specimens from younger subjects were not significantly different to those in older subjects. CONCLUSIONS: Collagen VI is a major component of the islet-exocrine interface of the adult pancreas, the content being more than double that of collagen I or IV. However, the proportional collagen VI content was not dependent on the age of the donor. These data may facilitate the design of new collagenases, targeting major substrates such as collagen VI in order to improve clinical islet isolation. PMID- 16477232 TI - The difficulty of eliminating donor leukocyte microchimerism in rat recipients bearing established organ allografts. AB - BACKGROUND: Unequivocal eradication of donor leukocyte microchimerism from recipients of long-surviving organ transplants has never been reported. Here we describe a drastic attempt to accomplish this objective. METHODS: In control experiments, a rank order of microchimerism and of associated donor specific nonreactivity was produced in Brown-Norway (BN) rats by transplantation of Lewis (LEW) liver, bone marrow cell (BMC) and heart allografts under a brief course of tacrolimus. The degree of microchimerism at 60 and 110 days was estimated with semiquanitative immunocytochemical and PCR techniques. Tolerance at 110 days was assessed in the different control groups by challenge transplantation of naive LEW hearts. In parallel experimental groups, an attempt was made to eliminate microchimerism from the BN recipients. The animals were submitted at 60 days to 9.5-Gy total body irradiation (TBI), reconstituted immediately with naive BN BMC, and tested for donor specific nonreactivity by LEW heart transplantation at 110 days. RESULTS: After the TBI-reconstitution at 60 days, microchimerism was undetectable in BMC recipients at 110 days, significantly reduced in heart recipients, and least affected in liver recipients. Except in liver recipients, abrogation of LEW-specific nonreactivity was demonstrated by rejection of the priming grafts, or by rejection of the challenge heart grafts, and by in vitro immune assay. CONCLUSIONS: It is difficult to eliminate microchimerism in organ recipients once the donor cells have settled into tissue niches. PMID- 16477233 TI - Rapamycin does not induce anergy but inhibits expansion and differentiation of alloreactive human T cells. AB - BACKGROUND: Studies in mice have shown that rapamycin inhibits cell cycle progression and promotes the development of clonal anergy. We here addressed the question if rapamycin can induce anergy of human T cells and studied the effects of rapamycin on activation, proliferation and expression of cytotoxic effector molecules of alloresponsive T cells in mixed lymphocyte cultures. METHODS: Peripheral blood mononuclear cells from healthy individuals were labeled with CFSE to monitor subsequent cell divisions. Cells were cocultured with allogeneic irradiated cells in the presence or absence of rapamycin. Flowcytometric analysis was performed after staining for surface CD4, CD8, and CD25 and for intracellular perforin, granzyme B, active caspase-3, and TGF-beta. Bio-Plex cytokine assay was done to measure the secretion of IL-2, IL-4, IL-10, and IFN-gamma. RESULTS: Addition of rapamycin at a final concentration of 10 ng/ml strongly decreased precursor frequencies of alloreactive CD4+ and CD8+ T cells. However, when these cells were washed and subsequently specifically restimulated in the absence of rapamycin, the proliferative capacity appeared normal. Next to lowering precursor frequencies, rapamycin also inhibited T cell expansion by inducing apoptosis in divided alloreactive CD4+ and CD8+ T cells. Rapamycin did not interfere with the formation of CD25brightCD4+ T cells during allogeneic stimulation and did not inhibit their suppressive function. Furthermore, the drug decreased production of effector molecules perforin and granzyme B by alloreactive T cells and diminished alloreactive cytotoxicity. CONCLUSION: Our data show that rapamycin strongly inhibits proliferation and effector functions of alloreactive T cells in vitro, but does not induce alloantigen specific nonresponsiveness. PMID- 16477231 TI - Prolonged survival of mice with acute liver failure with transplantation of monkey hepatocytes cultured with an antiapoptotic pentapeptide V5. AB - BACKGROUND: Because hepatocyte transplantation has been considered to be an attractive method to treat acute liver failure (ALF), efficient recovery of hepatocytes and maintenance of differentiated hepatocyte functions is of extreme importance. We here report the usefulness of an antiapoptotic pentapeptide V5, composed of Val-Pro-Met-Leu-Lys, in the monkey hepatocyte cultures. METHODS: We evaluated albumin production, metabolizing abilities of ammonia, lidocaine, and diazepam of monkey hepatocytes cultured with V5. The protein expression of apoptosis-associated molecules was analyzed using power blot analysis. An unwoven cloth inoculated with V5-treated monkey hepatocytes was transplanted on the surface of the spleen of both SCID mice and Balb/c mice suffering from ALF induced by 90% hepatectomy. RESULTS: When 100 microM V5 was utilized, ammonia-, lidocaine- and diazepam- metabolizing capacities and albumin production ability were significantly increased in V5-treated monkey hepatocytes. Such hepatocytes showed decreased Annexin V binding and increased the expression of anti-apoptotic and/or cytoprotective molecules, including Ku70, NF-kappaB, IKAP, hILP/XIAP, IkappaB, and CAS. Transplantation of the cloth containing the monkey hepatocytes significantly improved blood levels of glucose and ammonia and encephalopathy score and prolonged the survival of the mice with ALF. CONCLUSIONS: The present work clearly demonstrates the usefulness of V5 for maintaining the functions of monkey hepatocytes in tissue culture. PMID- 16477234 TI - Immunosuppressive and trafficking properties of donor splenic and bone marrow dendritic cells. AB - BACKGROUND: Infusion of donor dendritic cells (DC) has been shown to prolong allograft survival in a number of models. However, many regimens that utilize donor DC do not consistently produced tolerance or long-term allograft survival. We hypothesized that one factor limiting the therapeutic effect of donor DC is their relative inability to traffic to recipient peripheral lymph nodes and inhibit the function of resident alloreactive T cells. METHODS: Donor strain DC isolated from the spleens or bone marrow of Flt3L-treated mice were transferred intravenously into recipients at the time of skin grafting. Where indicated, recipients were treated with an anti-CD40L antibody and CTLA4-Ig. RESULTS: Infusion of donor DC together with costimulatory blockade promoted donor-specific prolongation of skin allograft survival in mice. Perhaps due to their more immature phenotype, bone marrow DC trafficked more effectively to the spleen, bone marrow, and thymus and were associated with significantly longer allograft survival than were splenic DC. Neither population of DC trafficked well to peripheral lymph nodes. Consistent with our hypothesis, splenic but not lymph node T cells from DC-treated recipients displayed donor-specific hyporesponsiveness in vitro. CONCLUSION: These data suggest that one factor contributing to rejection following treatment with donor DC plus costimulation blockade is the persistence of donor-reactive T cells within the recipient's secondary lymphoid structures. Strategies to improve DC trafficking to these structures may enhance their therapeutic effect. PMID- 16477235 TI - ACE gene D/D genotype as a risk factor for chronic nephrotoxicity from calcineurin inhibitors in liver transplant recipients. AB - BACKGROUND: Chronic renal failure (CRF) is a major cause of morbidity and mortality after orthotopic liver transplantation (OLTX) and is predominantly caused by calcineurin inhibitors (CI)-induced nephrotoxicity. The activation of the renin angiotensin system (RAS) has been implicated in the pathogenesis of chronic nephrotoxicity from CI. METHODS: We retrospectively investigated the genes coding for components of the RAS (ACE gene, Angiotensin II receptor 1 gene, Angiotensinogen gene) in 233 liver transplant recipients receiving Cyclosporine (CsA) or Tacrolimus (Tac) as maintenance immunosuppressant. All patients with serum creatinine (sCr) <1.0 mg/dL (n=143) before orthotopic liver transplantation (OLTX) were included in the final analysis. Patients were than categorized into two groups based upon their most recent postliver transplant sCr level: Group 1 (n=83) with sCr <1.5 mg/dL (mean 1.1+/-0.2) and group 2 (n=60) with sCr > or =1.5 mg/dL (mean 2.5+/-1.3) RESULTS: ACE D/D genotype was found in 57% of patients with sCr > or =1.5 mg/dL compared to 20% of patients with sCr <1.5 mg/dL (P<0.0001) CONCLUSIONS: Our analysis strongly suggests that liver transplant patients with ACE gene D/D genotype are at a significant higher risk of developing CI-induced chronic nephrotoxicity. PMID- 16477236 TI - Prediction of the glomerular filtration rate using equations in kidney-pancreas transplant patients receiving cimetidine. AB - BACKGROUND: Current estimates of renal function in kidney transplant patients are frequently inaccurate compared to radionuclide GFR (rGFR) measurement. Cimetidine inhibits tubular secretion of creatinine and improves the accuracy of formulas to estimate GFR. METHODS: We studied the effect of a cimetidine-aided (800 mg tid for 24 hr) serum creatinine on the correlation of creatinine clearance and three prediction formulas (Cockroft-Gault, Levey, and Nankivell) compared to rGFR in 15 kidney-pancreas transplant patients. Results were adjusted for body surface area. RESULTS: Correlations with rGFR using cimetidine-aided creatinine were: Cockroft Gault, r=0.710; Levey, r=0.752; Nankivell, r=0.676; creatinine clearance, r=0.643. By Bland and Altman analysis, agreement with rGFR was best with the Nankivell and Cockroft-Gault equations and worst with creatinine clearance. Cimetidine ($0.48 Canadian) costs substantially less than the rGFR test ($66.00 Canadian). CONCLUSION: Using cimetidine, prediction equations give a stronger correlation with GFR than creatinine clearance. PMID- 16477237 TI - A novel strategy for corneal endothelial reconstruction with a bioengineered cell sheet. AB - Cellular organization of foreign grafts constructed from cultivated cells is critical to successful graft-host integration and tissue repair. This study described a novel human corneal endothelial cell (HCEC) therapeutic method, where cultivated adult HCEC sheet with uniform orientation was prepared and transplanted to a rabbit cornea. Having a correct morphology and intact barriers, the HCEC sheet was made by the temperature-modulated detachment of monolayered HCECs from thermoresponsive poly(N-isopropylacrylamide) (PNIPAAm)-grafted surfaces and was delivered with proper polarity to the corneal posterior surface by a bioadhesive gelatin disc. Results of the in vivo studies, including the follow-up clinical observations and histological examinations, showed the laminated HCEC sheet was successfully integrated into rabbit cornea denuded with endothelial layer after the biodegradation of gelatin carrier. These data indicate the feasibility of the proposed procedure in cell therapy for corneal endothelial cell loss. PMID- 16477238 TI - Prevalence of antibodies to Trypanosoma cruzi among solid organ donors in Southern California: a population at risk. AB - Trypanosoma cruzi, a parasite that causes Chagas' disease, is endemic in parts of Mexico, South America, and Central America. Transmission of T. cruzi infection by solid organ transplantation has been reported in Latin America and recently in the United States. To determine the prevalence of T. cruzi antibodies in Southern California organ donors, 404 samples from deceased organ donors between May 2002 to April 2004 were screened using a qualitative enzyme-linked immunosorbent assay (EIA) and confirmed with an immunofluorescence assay (IFA) available through the Centers for Disease Control (CDC). Six donors were initially reactive by EIA. Three donors were repeatedly reactive after repeat testing and were sent to the CDC for confirmation. One donor (0.25%) had an IFA-confirmed reactivity to anti T. cruzi antibodies. In areas where there is a high number of immigrants from T. cruzi endemic countries, screening for anti-T. cruzi donor antibodies may be beneficial. PMID- 16477239 TI - Comparison of histidine-tryptophan ketoglutarate solution and University of Wisconsin solution in prolonged cold preservation of kidney allografts. AB - Although University of Wisconsin (UW) solution is the standard preservation solution for organ transplantation, Histidine-Tryptophan Ketogluatarate (HTK) solution has been increasingly used. This study compared HTK or UW for cold static storage of kidney allografts. In all, 149 renal transplants were performed with cold ischemic times (CI) greater than 16 hr (UW 87, HTK 62) and a subset analysis was performed with CI over 24 hr (HTK 31, UW 38). Data from receiving renal transplant centers focused on delayed graft function (DGF), patient and allograft survival. In CI greater than 16 hr, graft and patient survival were comparable. HTK cohort had lower DGF. In CI greater than 24 hr, there was no difference in patient survival, a trend towards improved graft survival in HTK, and decreased rate of DGF in HTK. This data suggests that UW and HTK have at least similar efficacy in kidney preservation at longer ischemic times. PMID- 16477241 TI - Interaction between tacrolimus and omeprazole in a pediatric liver transplant recipient. PMID- 16477242 TI - Induction of autoimmune hepatitis by pegylated interferon in a liver transplant patient with recurrent hepatitis C virus. PMID- 16477240 TI - Real-time direct measurement of human liver allograft temperature from recovery to transplantation. AB - Temperature is a key parameter in organ preservation that has been consistently linked to primary nonfunction (PNF). In this communication, and for the first time anywhere, continued and direct measurements of human liver intraparenchymal temperatures are reported in six clinical cases of orthotopic liver transplantations (OLT). These measurements cover the entire transplantation procedure and include the full transport phase. In contrast with long-held beliefs, these data demonstrate that liver allograft temperatures reach and stabilize at near 0 degrees C, instead of 4 degrees C, during transport using standard protocols. Furthermore, these low temperatures do not appear to contribute to graft failure when negative factors such as long preservation, the presence of hepatic steatosis, or advanced donor age are present. The clinical and experimental implications of these findings, together with other relevant elements derived from the direct and continuous monitoring of human liver allograft intraparenchymal temperatures, are discussed. PMID- 16477243 TI - Liver-occupying focal nodular hyperplasia and adenomatosis associated with intrahepatic portal vein agenesis requiring orthotopic liver transplantation. PMID- 16477245 TI - Oxygen and pancreas preservation. PMID- 16477246 TI - Localized intraocular posttransplant lymphoproliferative disorder after pediatric liver transplantation. PMID- 16477248 TI - Epidemiologic research in the face of an obesity epidemic. PMID- 16477249 TI - The garden of Eden: plant-based diets, the genetic drive to store fat and conserve cholesterol, and implications for epidemiology in the 21st century. PMID- 16477250 TI - Epidemiologic research on the obesity epidemic: a socioenvironmental perspective. PMID- 16477251 TI - Methodologic issues for the study of obesity. PMID- 16477252 TI - Prone to sleep, prone to pyloric stenosis? PMID- 16477253 TI - In utero and postnatal maternal smoking and asthma in adolescence. AB - BACKGROUND: Asthma in early childhood has been associated with maternal smoking during pregnancy and parental smoking soon after birth. However, less is known about these exposures and the development of asthma symptoms in adolescence. METHODS: Data were taken from the Mater University Study of Pregnancy, a large birth cohort study of mothers and children enrolled in Brisbane, Australia, beginning in 1981. Smoking was assessed at 2 stages during pregnancy and at the 6 month and 5-year follow-up visits. Asthma was assessed from maternal reports that were provided when the child was age 14 years. We conducted multivariable multinomial logistic regression analyses to assess the effect of maternal smoking on asthma symptoms. RESULTS: There was a strong sex interaction such that girls whose mothers had smoked heavily (20 or more cigarettes per day) in pregnancy and at the 6-month follow up had increased odds of experiencing asthma symptoms at age 14 (odds ratio = 1.96; 95% confidence interval = 1.25-3.08). The contribution of heavy smoking during pregnancy appeared to be stronger than heavy smoking after the birth. No similar associations were seen for boys. CONCLUSION: Female adolescents whose mothers smoked heavily during the fetal period and the early months of life have increased risk of asthma symptoms in adolescence. In utero exposure to heavy smoking was found to have a stronger effect than postnatal environmental tobacco exposure. PMID- 16477254 TI - Estimating disease prevalence in a Bayesian framework using probabilistic constraints. AB - Studies sometimes estimate the prevalence of a disease from the results of one or more diagnostic tests that are applied to individuals of unknown disease status. This approach invariably means that, in the absence of a gold standard and without external constraints, more parameters must be estimated than the data permit. Two assumptions are regularly made in the literature, namely that the test characteristics (sensitivity and specificity) are constant over populations and the tests are conditionally independent given the true disease status. These assumptions have been criticized recently as being unrealistic. Nevertheless, to estimate the prevalence, some restrictions on the parameter estimates need to be imposed. We consider 2 types of restrictions: deterministic and probabilistic restrictions, the latter arising in a Bayesian framework when expert knowledge is available. Furthermore, we consider 2 possible parameterizations allowing incorporation of these restrictions. The first is an extension of the approach of Gardner et al and Dendukuri and Joseph to more than 2 diagnostic tests and assuming conditional dependence. We argue that this system of equations is difficult to combine with expert opinions. The second approach, based on conditional probabilities, looks more promising, and we develop this approach in a Bayesian context. To evaluate the combination of data with the (deterministic and probabilistic) constraints, we apply the recently developed Deviance Information Criterion and effective number of parameters estimated (pD) together with an appropriate Bayesian P value. We illustrate our approach using data collected in a study on the prevalence of porcine cysticercosis with verification from external data. PMID- 16477255 TI - Estimated effects of hydrazine exposure on cancer incidence and mortality in aerospace workers. AB - BACKGROUND: Animal studies suggest that hydrazine is a lung carcinogen, but human studies have been rare, rather small, and limited to cancer mortality. METHODS: We examined cancer mortality and incidence in a cohort of aerospace workers with varying exposure to hydrazine contained in rocket fuels-extending previous mortality follow-up from 1994 to 2001 and investigating cancer incidence for the period 1988-2000 using population-registry data. We newly estimated hydrazine effects adjusting for occupational exposures to other carcinogens assessed through a job-exposure matrix. Rate-ratio estimates were derived from Cox proportional hazards and random-effects models using time-dependent exposure measures for hydrazine adjusting for trichloroethylene, polycyclic aromatic hydrocarbons, benzene, and mineral oil exposures. RESULTS: Exposure to hydrazine was positively associated with lung cancer incidence (estimated rate ratio for high vs low exposure with 20-year lag = 2.5; 95% confidence interval = 1.3-4.9) and with colorectal cancer incidence (2.2; 1.0-4.6). Dose-response associations were observed for both outcomes; similar associations were found for lung cancer mortality but not for colorectal cancer mortality. Effect estimates for cancers of the pancreas, blood and lymph system, and kidneys were based on small numbers rendering our analyses uninformative, and patterns considering exposure levels and lags were inconsistent. Use of random-effect models did not change our results. CONCLUSIONS: The findings reported here are consistent with our previous results for lung cancer mortality; our new results suggest that exposure to hydrazine increases the risk of incident lung cancers. We also found, for the first time, an increased risk of colon cancers. Results for other cancer sites are inconclusive. PMID- 16477256 TI - Mortality rates among augmentation mammoplasty patients: an update. AB - BACKGROUND: A large follow-up study of cosmetic breast implant patients previously suggested an overall decrease in mortality but increased risks of brain and respiratory cancers and of suicides. METHODS: This cohort of 12,144 implant patients and 3614 patients with other types of plastic surgeries was followed for 5 additional years, enabling derivations of standardized mortality ratios (SMRs) based on population rates and relative risks (RRs) based on comparisons with the other patients. RESULTS: A total of 443 implant and 221 other plastic surgery patients were identified as deceased (SMR = 0.65 [95% confidence interval (CI) = 0.6-0.7] and 0.56 [0.5-0.6], respectively). Despite evidence that implants can interfere with mammographic visualization, there was no evidence that implant patients had a higher risk of death from breast cancer as compared with either the general population or other plastic surgery patients. The previous excess risk of brain cancer deaths among implant patients was attenuated by follow-up (as the result of no additional deaths; SMR = 1.43, 0.8 2.5; RR = 2.07, 0.5-8.9). A previously observed excess risk of respiratory cancer deaths persisted in comparisons with other plastic surgery patients (RR = 1.63; 1.0-2.7), but there was no evidence of a trend of risk with follow-up time. Implant patients also showed an elevated risk of suicide (SMR = 1.63, 1.1-2.3; RR = 2.58, 0.9-7.8) and of deaths caused by motor vehicle accidents (RR = 1.73; 0.6 5.4). CONCLUSIONS: Although several elevations in cause-specific mortality were attenuated by additional follow-up, the excess risk of suicide among the implant patients remains of concern. PMID- 16477257 TI - Combined effects of prepregnancy body mass index and weight gain during pregnancy on the risk of preterm delivery. AB - BACKGROUND: The association between excessive gestational weight gain and preterm delivery is unclear, as is the association between low gestational weight gain and preterm delivery among overweight and obese women. METHODS: Using data from the Pregnancy Risk Assessment Monitoring System in 21 states, we estimated the risk of very (20-31 weeks) and moderately (32-36 weeks) preterm delivery associated with a combination of prepregnancy body mass index (BMI) and gestational weight gain among 113,019 women who delivered a singleton infant during 1996-2001. We categorized average weight gain (kilograms per week) as very low (<0.12), low (0.12-0.22), moderate (0.23-0.68), high (0.69-0.79), or very high (>0.79). We categorized prepregnancy BMI (kg/m) as underweight (<19.8), normal (19.8-26.0), overweight (26.1-28.9), obese (29.0-34.9), or very obese (>or=35.0). We examined associations for all women and for all women with no complications adjusting for covariates. RESULTS: There was a strong association between very low weight gain and very preterm delivery that varied by prepregnancy BMI, with the strongest association among underweight women (adjusted odds ratio = 9.8; 95% confidence interval = 7.0-13.8) and the weakest among very obese women (2.3; 1.8-3.1). Very low weight gain was not associated with moderately preterm delivery for overweight or obese women. Women with very high weight gain had approximately twice the odds of very preterm delivery, regardless of prepregnancy BMI. CONCLUSIONS: This study supports concerns about very low weight gain during pregnancy, even among overweight and obese women, and also suggests that high weight gain, regardless of prepregnancy BMI, deserves further investigation. PMID- 16477258 TI - Race, cardiovascular reactivity, and preterm delivery among active-duty military women. AB - BACKGROUND: Rates of preterm delivery in the United States are higher in black women compared with whites. In this study, we examined cardiovascular reactivity and risk of preterm delivery among black and white military women. METHODS: We recruited a total of 500 black and white active-duty military women from the prenatal clinic at a large military installation, interviewing them early in pregnancy and again at 28 weeks of gestation. A subgroup of women underwent a computerized stress test to determine cardiovascular reactivity assessed as increases in heart rate and blood pressure compared with measurements taken before the stress test. RESULTS: Despite a relatively low overall risk of preterm delivery (8.2%), we found the same 2-fold racial disparity reported in other populations (hazard ratio for preterm delivery in black women vs whites = 2.30; 95% confidence interval = 1.24-4.27). The disparity is present in all military ranks and is largest for medically indicated preterm deliveries. Among the 313 subjects who participated in the computerized stress testing, blacks exhibited more cardiac reactivity than whites. In black subjects only, a 1-mm increase in diastolic blood pressure reactivity was associated with 1.1 a day earlier delivery (-0.17 weeks). A similar trend was seen with heart rate. CONCLUSIONS: Autonomic dysfunction after exposure to stressors may play a role in the timing of delivery among black women. PMID- 16477259 TI - High-sensitivity C-reactive protein and cognitive function in older women. AB - BACKGROUND: Inflammatory processes may be involved in the development of dementia, although findings from epidemiologic studies directly examining inflammatory markers and dementia or its precursor, impaired cognitive function, are inconsistent. METHODS: We measured plasma levels of the inflammatory marker, C-reactive protein, using a high-sensitivity assay (hs-CRP) in 4,231 older participants of the Women's Health Study, who provided blood samples between 1992 and 1996 when they were age 60 to 90 years. From 1998 to 2000, we administered a battery of 5 cognitive tests measuring general cognition, verbal memory, and category fluency. Using multiple linear regression, we compared mean cognitive test scores across quintiles of hs-CRP, adjusting for potential confounding factors. RESULTS: There was a wide distribution of hs-CRP levels among these women, and a large proportion had levels considered to reflect a high risk of cardiovascular disease. We observed no suggestion, however, that higher hs-CRP levels were associated with poorer cognitive performance. For example, on a global score combining results of all the cognitive tests, mean scores among women in the highest quintile of hs-CRP did not differ from those in the lowest quintile (multivariable-adjusted mean difference = 0.04; 95% confidence interval [CI]: -0.02 to 0.11, P for trend across quintiles = 0.38). CONCLUSION: Overall, in these women, we found no evidence of a link between hs-CRP, a marker of inflammation, and decrements in cognitive function. PMID- 16477260 TI - No association between telomere length and survival among the elderly and oldest old. AB - BACKGROUND: The consistent findings of a negative correlation between telomere length and replicative potential of cultured cells, as well as a decreasing telomere length in a number of different tissues in humans with age, have led to the suggestion that telomeres play a role in cellular aging in vivo and ultimately even in organismal aging. Furthermore, one small longitudinal study of elderly individuals has suggested that longer telomeres are associated with better survival. METHODS: Telomere length was measured as mean terminal restriction fragment length on blood cells from 812 persons, age 73 to 101 years, who participated in population-based surveys in 1997-1998. Among the participants were 652 twins. The participants were followed up through the Danish Civil Registration system until January 2005, at which time 412 (51%) were dead. RESULTS: Univariate Cox regression analyses revealed that longer telomeres were associated with better survival (hazard ratios = 0.89 [95% confidence interval = 0.76-1.04] per 1 kb in males and 0.79 [0.72-0.88] per 1 kb in females, respectively). However, including age in the analyses changed the estimates to 0.97 (0.83-1.14) and 0.93 (0.85-1.03), respectively. Intrapair comparison showed that among 175 twin pairs in which at least one died during follow up, it was the twin with the shorter telomere length who died first in 97 (55%) of the pairs (95% confidence interval = 48-63%). We could not confirm the recently reported negative correlation between telomere length and obesity or between telomere length and smoking. CONCLUSION: This longitudinal study of the elderly and oldest old does not support the hypothesis that telomere length is a predictor for remaining lifespan once age is controlled for. PMID- 16477261 TI - The continuing enigma of pyloric stenosis of infancy: a review. AB - Striking features of the descriptive epidemiology of pyloric stenosis of infancy have been identified but until recently have not suggested any useful etiologic lead. The disease first received serious attention approximately 100 years ago and has since occurred throughout the Western world at a rate between 2 and 5 per thousand live births; it appears to be uncommon elsewhere. Its age distribution is essentially limited to the period between the third and eighth weeks after birth. It is 4 to 5 times more common in boys than girls. It is less common in blacks than whites in the United States and less common among Asians than whites in the United States and elsewhere. Its incidence is highest in first-born infants. Evidence on a role for maternal age is not consistent. The disease re occurs in families with sufficient frequency to incite the interest of geneticists, although no genetic model yet proposed offers a better basis for counseling than do the empiric observations on which it is based. Monozygous twins are concordant for the disease not much more frequently than are dizygous twins, and indeed not a great deal more often than nontwin siblings, which should prompt a search for environmental explanations of the disease's familial nature. Sharp declines in the incidence of the disease in Denmark and Sweden during the 1990s led to the hypothesis that infants sleeping in the prone position-a practice discouraged with some success by Scandinavian campaigns to reduce the frequency of sudden infant death syndrome (SIDS)-may also be at increased risk of pyloric stenosis. If supported, this hypothesis may offer the first-ever possibility of reducing the frequency of this disease, as well as SIDS. If the hypothesis is not supported, the recent declines in the disease in Denmark and Sweden add another facet to its enigmatic nature. PMID- 16477262 TI - Autoimmune diseases co-occurring within individuals and within families: a systematic review. AB - BACKGROUND: Autoimmune diseases have been observed to coexist both within individuals and within families. It is unclear whether clinical reports of comorbid autoimmune diseases represent chance findings or true associations. This systematic review evaluates the current level of evidence on the coexistence of selected autoimmune diseases within individuals and families. We reviewed the associations among 4 TH1-associated autoimmune diseases: insulin-dependent diabetes mellitus, autoimmune (Hashimoto) thyroiditis, rheumatoid arthritis, and multiple sclerosis. METHODS: Studies quantifying the coexistence between the selected diseases, published through March 2004, were identified from Medline and Embase searches. Study eligibility was determined on the basis of preestablished criteria, and relevant data were extracted according to a fixed protocol. We determined the prevalence of comorbid autoimmune disease according to index disease and then compiled summary statistics. Heterogeneity among studies was assessed by exact likelihood ratio tests and Monte Carlo inference. RESULTS: We found 54 studies that met the eligibility criteria. Of these, 52 studies examined the coexistence of disease within individuals and 9 studies examined within family associations. The majority of studies were uncontrolled and did not account for confounding factors. There was substantial evidence for heterogeneity among studies. Although inconclusive, the data appear to support an increased prevalence of autoimmune thyroiditis among patients with rheumatoid arthritis and those with insulin-dependent diabetes mellitus, and an inverse association between rheumatoid arthritis and multiple sclerosis. CONCLUSION: Although the available evidence does not permit firm conclusions regarding comorbidities among the selected autoimmune diseases, results are sufficiently suggestive to warrant further study. PMID- 16477263 TI - Paternal age and preterm birth in Italy, 1990 to 1998. AB - BACKGROUND: Advanced paternal age has been reported to impair pregnancy outcome. Here, we investigated the association of advanced paternal age with preterm birth by using a very large national data set. METHODS: We analyzed data from 1990 to 1998 on Italian firstborn singletons to mothers 20-24 and 25-29 years of age (n = 1,510,823). Odds ratios for overall preterm (<37 weeks' gestation), very preterm (<32 weeks), and moderate preterm (32-36 weeks) births were evaluated through logistic regression models in paternal age classes (20-24, 25-29, 30-34, 35-39, 40-44, 45-49, 50+ years) after adjustment for confounders. Nonparametric regression models were used to fit the effect of paternal ageing on the incidence of very preterm births. RESULTS: Odds ratios increased with paternal age more rapidly for very preterm than for moderate preterm births; among 45- to 49-year old fathers, odds ratios for very preterm births reached 1.91 (95% confidence interval = 1.08-3.38) and 1.72 (1.25-2.36), respectively, in 20- to 24- and 25- to 29-year-old mothers. CONCLUSIONS: This study confirms that paternal age contributes to the risk of preterm birth. The effect is stronger on very preterm births but also influences moderate preterm births. PMID- 16477264 TI - Physical activity and magnetic field exposure in pregnancy. AB - BACKGROUND: Peak magnetic field exposure was associated with increased risk of miscarriage in 2 recent studies. Reduced physical activity levels in healthy pregnancies may affect measured exposure and thus bias results. METHODS: We recruited 100 pregnant women to wear an Actigraph accelerometer and EMDEX magnetic field monitor for a 7-day period. We evaluated the association between physical activity and magnetic field exposure (peaks and time-weighted average) using generalized estimating equations and linear mixed models. RESULTS: We found a positive association between level of activity and likelihood of incurring elevated exposure in the person-day analysis, most strongly for cutpoints of 16 or 20 mG, for both working and nonworking women among whom odds ratios in the uppermost quartile ranged from 2.1 to 2.6. A positive association was found using person-minutes only among nonworking women. CONCLUSION: Physical activity may affect peak magnetic field exposure. If the early nausea and later cumbersomeness of healthy pregnancies leads to reduced physical activity, this could distort measured magnetic field-health outcome associations. PMID- 16477265 TI - Validation of adolescent diet recalled by adults. AB - BACKGROUND: Few studies have examined the validity of adolescent diet as recalled by adults many years later. METHODS: In this investigation, we examined the validity of a 124-item food frequency questionnaire (HS-FFQ) completed by 80 young adults in the United States about their diet during high school. We compared the HS-FFQ with 3 24-hour recalls and 2 food-frequency questionnaires (YAQ) collected 10 years earlier in 1993, when the participants were in high school. We calculated Pearson correlation coefficients for 20 to 25 nutrients and corrected these correlations for within-person variation. RESULTS: The average corrected correlation for energy-adjusted nutrient intakes calculated from the HS FFQ and the 24-hour recalls was 0.45 (range = 0.16-0.68). The mean corrected correlation between the HS-FFQ and YAQs was 0.58 (range = 0.40-0.88). CONCLUSION: Taken together with the results of our earlier work, this analysis suggests that the HS-FFQ can reasonably capture adolescent diet recalled by young adults. PMID- 16477266 TI - Short-term effects of ambient particles on cardiovascular and respiratory mortality. AB - BACKGROUND: Particulate air pollution is associated with increased mortality. There is a need for European results from multicountry databases concerning cause specific mortality to obtain more accurate effect estimates. METHODS: We report the estimated effects of ambient particle concentrations (black smoke and particulate matter less than 10 mum [PM10]) on cardiovascular and respiratory mortality, from 29 European cities, within the Air Pollution and Health: a European Approach (APHEA2) project. We applied a 2-stage hierarchical modeling approach assessing city-specific effects first and then overall effects. City characteristics were considered as potential effect modifiers. RESULTS: An increase in PM10 by 10 microg/m (lag 0 + 1) was associated with increases of 0.76% (95% confidence interval = 0.47 to 1.05%) in cardiovascular deaths and 0.58% (0.21 to 0.95%) in respiratory deaths. The same increase in black smoke was associated with increases of 0.62% (0.35 to 0.90%) and 0.84% (0.11 to 1.57%), respectively. CONCLUSIONS: These effect estimates are appropriate for health impact assessment and standard-setting procedures. PMID- 16477267 TI - Causes and mechanisms: an interview with Jeremiah Stamler. PMID- 16477269 TI - Human fertility decline? PMID- 16477271 TI - Resting energy expenditure and fuel metabolism following laparoscopic adjustable gastric banding in severely obese women: relationships with excess weight lost. AB - BACKGROUND: Although adjustable gastric banding is increasingly proposed for massively obese patients, little is known about the modifications of resting metabolic rate and substrate oxidation or about metabolic determinants of weight loss following this type of bariatric surgery. OBJECTIVES: To evaluate the relationships between excess weight loss, resting metabolic rate (RMR) and substrate oxidation, and to identify metabolic predictive factors of weight loss after adjustable gastric banding. SUBJECTS: Seventy-three obese nondiabetic women aged 39.1+/-10.4 years (18.4-64.8). DESIGN: Resting metabolic rate and substrate oxidation (indirect calorimetry), body composition (bio-impedance), lipid profile and insulin sensitivity indexes were assessed before and after (13.3+/-6.0 months, range 6.0-31.1) adjustable gastric banding. Patients were classified according to postsurgery time: group A (6-12 months, n=39); group B (12-18 months, n=21); group C (>18 months, n=13). Metabolic parameters associated with the percentage of excess weight lost (EWL) 1 year after surgery were analyzed in univariate and multivariate regressions. RESULTS: Mean weight loss was 26.2+/ 11.4 kg. Mean fat mass loss was 17.3+/-8.1 kg. All biological parameters associated with excess weight improved after surgery. Excess weight lost at 1 year was 45.9+/-17.1% in group A, 47.4+/-17.1% in group B and 51.4+/-18.5% in group C (P=NS). Resting metabolic rate/fat-free mass (FFM) slightly decreased (28.9+/-3.26 vs 30.3+/-2.8, P<0.00001) and RMR/body weight slightly increased (18.5+/-2.8 vs 17.3+/-1.9, P<0.00001) after surgery. Respiratory quotient (0.81+/ 0.06 vs 0.82+/-0.05) and FFM-adjusted lipid oxidation (1.10+/-0.41 vs 1.05+/-0.33 mg/min/kg FFM) were not significantly modified after surgery. In multiple linear regression analysis, difference in RMR/body weight, difference in energy sparing, baseline BMI and postsurgery time, were significantly and independently correlated with EWL (total R2=72.5%). CONCLUSIONS: Adjustable gastric banding promotes gradual but sustained weight loss and is associated with long-term conservation of lipid oxidation and energy expenditure. The individual variability in energy sparing mechanisms predicts weight loss during the first year after surgery. PMID- 16477270 TI - Alleviation of hepatic steatosis accompanied by modulation of plasma and liver TNF-alpha levels by Trigonella foenum graecum (fenugreek) seeds in Zucker obese (fa/fa) rats. AB - OBJECTIVE: Oral supplements of fenugreek (Trigonella foenum graecum) seeds (Fen) have been shown to treat glucose and lipid homeostasis in several metabolic disorders; however, its ability to alleviate obesity-associated pathologies is not known. The main objective of this study was to evaluate the effect of Fen in Zucker obese rats (Ob), an animal model of obesity and related disorders, such as dyslipidemia and hepatic steatosis. METHODS: Female Zucker (6-week-old) Ob and lean (Ln) rats were randomly grouped (n = 8 rats/group) to receive either basal or 5% Fen-supplemented AIN-93G diets for 8 weeks, and then were euthanized. Histopathology and biochemical parameters in the liver together with plasma biochemistry were assessed. RESULTS: Obese rats had significantly higher (P < 0.05) body and liver weight, as well as plasma insulin, lactate, cholesterol, triglyceride and tumor necrosis factor (TNF)-alpha compared to their Ln counterparts. Fen significantly reduced (P < 0.05) the liver-weight of Ob rats in comparison to Ob rats fed basal diet (Control), while no effect was observed in the Ln rats. Fen treatment resulted in a lower P-value (P < 0.05). In addition, Ob rats on Fen-supplemented diets had fasting plama cholesterol and TNF-alpha levels, and significantly higher (P < 0.05) triglycerides in comparison to the control rats. Hepatic triglyceride level was significantly lower (P < 0.05) in Ob rats fed Fen supplemented diets in comparison to control. The levels of bound and soluble liver TNF-alpha (26 and 17 kDa, respectively) and TNF receptor-II (TNFR II, 75 kDa) proteins were significantly lower (P < 0.05) in Ob than in Ln rats. Fen significantly lowered (P < 0.05) both the soluble and bound forms of TNF alpha protein while significantly elevating (P < 0.05) TNFR-II in the livers of Ob rats compared to Ob Controls. CONCLUSION: These results demonstrate unequivocally that in a short-term preclinical evaluation, dietary Fen supplementation reduced the triglyceride accumulation in the liver, a hallmark feature of hepatic steatosis without affecting the plasma insulin or glucose levels in Zucker obese rats and suggest that TNF-alpha may play an important role in this process. PMID- 16477272 TI - Increase in serum resistin during weight loss in overweight subjects is related to lipid metabolism. AB - OBJECTIVE: Human resistin has been stated to influence preadipocyte cell numbers and to stimulate adipocyte triglyceride lipolysis in vivo and in vitro. However, its role in human obesity remains unclear. DESIGN: Cross-sectional study for comparisons of lean and obese subjects, and subsequent longitudinal 4-month weight loss intervention study in obese subjects. SUBJECTS: Healthy subjects, lean (n=20, BMI<25) and overweight (n=43, BMI>or=25). MEASUREMENTS: Serum resistin, body weight, body fat, waist-to-hip ratio, as well as markers of insulin resistance and lipid metabolism at baseline and after 4 months of intervention. RESULTS: Serum resistin was positively correlated to HOMA-IR (partial r=0.288; P=0.055), serum fructosamines (partial r=0.280; P=0.062), serum NEFA (partial r=0.276; P=0.066) and negatively to age (partial r=-0.349; P=0.019) and serum apolipoprotein A-1 (partial r=-0.363; P=0.014). During the intervention, serum resistin increased significantly (P<0.001). The increase was inversely related to changes in waist-to-hip ratio (P=0.025) and positively to serum apolipoprotein B (P=0.011). In males only, the increase in resistin during weight loss was predicted by total serum cholesterol at baseline (r=0.703, P=0.007). No relation was observed between changes in resistin and changes in HOMA-IR. CONCLUSION: The present study indicates an association between serum resistin and markers of abdominal fat distribution as well as the regulation of lipid metabolism. However, human resistin is unlikely to play an independent role in the regulation of glucose metabolism. PMID- 16477273 TI - Variability of appetite control mechanisms in response to 9 weeks of progressive overfeeding in humans. AB - BACKGROUND: The current epidemic of obesity demonstrates that mechanisms for maintaining human energy balance are readily subverted by adverse environmental conditions. The critical elements of this dysregulation are poorly understood. Most previous research into what regulates the intake side of the energy balance equation has been handicapped by the use of short-term within-day experimental tests. OBJECTIVE: We enrolled six non-obese men to a 17-week protocol involving three 21 days periods of progressive overfeeding (+20, +40 and +60%) separated by free diet periods to test for compensatory satiety. RESULTS: Responses to overfeeding differed markedly with evidence of 'compensators' and 'non compensators', but on average, subsequent food intake was stimulated rather than suppressed after overfeeding in spite of markedly elevated body fat (+13%) and fasting leptin (+116%). DISCUSSION: The inefficient response of in-built appetite control mechanisms emphasizes the need to adopt intentional cognitive restraint in the modern environment when food is plentiful. PMID- 16477274 TI - Discrete opioid gene expression impairment in the human fetal brain associated with maternal marijuana use. AB - Fetal development is a period sensitive to environmental influences such as maternal drug use. The most commonly used illicit drug by pregnant women is marijuana. The present study investigated the effects of in utero marijuana exposure on expression levels of opioid-related genes in the human fetal forebrain in light of the strong interaction between the cannabinoid and opioid systems. The study group consisted of 42 midgestation fetuses from saline-induced voluntary abortions. The opioid peptide precursors (preprodynorphin and preproenkephalin (PENK)) and receptor (mu, kappa and delta) mRNA expression were assessed in distinct brain regions. The effect of prenatal cannabis exposure was analyzed by multiple regression controlling for confounding variables (maternal alcohol and cigarette use, fetal age, sex, growth measure and post-mortem interval). Prenatal cannabis exposure was significantly associated with increased mu receptor expression in the amygdala, reduced kappa receptor mRNA in mediodorsal thalamic nucleus and reduced preproenkephalin expression in the caudal putamen. Prenatal alcohol exposure primarily influenced the kappa receptor mRNA with reduced levels in the amygdala, claustrum, putamen and insula cortex. No significant effect of prenatal nicotine exposure could be discerned in the present study group. These results indicate that maternal cannabis and alcohol exposure during pregnancy differentially impair opioid-related genes in distinct brain circuits that may have long-term effects on cognitive and emotional behaviors. PMID- 16477275 TI - [Authorship and co-authorship]. PMID- 16477276 TI - [Palliative sedation--problem or obligation?]. PMID- 16477277 TI - [Generic name or brand name?]. PMID- 16477278 TI - [Low use of drugs among farmers]. AB - BACKGROUND: Farmers have a higher prevalence of depression compared to workers with other occupations. We wanted to investigate whether this corresponds to a more frequent use of antidepressants among farmers and to investigate their use of medication in general. MATERIAL AND METHOD: The study population consisted of 20 166 workers aged 40-47 years from the general population, including 398 farmers and 713 part-time farmers, from a population-based health study carried out in a Norwegian county. In addition to type of occupation and use of medical drugs, mental (HADS) and physical (SF-12) health, life-style factors, height, weight and blood pressure were measured. RESULTS: The farmers reported significantly lower use of antiperessants and also a significant lower use of medical drugs in general compared with other groups. Their physical health was significantly better compared with other occupational groups and they had a lower consumption of alcohol and tobacco. There were no marked differences in the blood pressure or body-mass index. CONCLUSION: The lower use of drugs could reflect better health, it could be related to a culturally-related reluctance to the use of drugs, or it could indicate that farmers in some instances are not receiving proper medical care. PMID- 16477279 TI - [A follow up of persons who received basic and/or supplemental benefits in childhood]. AB - BACKGROUND: Consequences of chronic diseases in childhood with respect to health, educational achievement and participation in the labour force in young adult age are evaluated. MATERIAL AND METHODS: A total of 14,364 children (2.3%) of the 626,928 born in Norway 1967-76 received basic and /or supplemental benefits for at least one year of the age span 0-16 years. The more common diagnoses included Endocrine diseases (diabetes), disease classified under Mental diagnoses, Neurological diseases and Congenital malformations. All the children were followed up to the age of 27 with respect to mortality and disability pensioning and to the age of 25 with regard to education, participation in the labour force and income, and in addition national service for the men. The study was made possible through the linking of data from several national registers, performed by Statistics Norway. Before the file was released for analyses, all personal identification was removed. RESULTS: Basic and supplementary benefits in childhood predict adverse outcomes in young adult age: mortality, disability, low education, lack of gainful employment and low pensionable income. The diagnosis registered with the benefit strongly influenced the outcomes. Conscripts who had received benefits were judged to have a mean score for general ability of 4.5 compared to 5.2. Adjusted for score for general ability the proportion of those having received benefits achieving higher education was 84% of that of those that had not received benefits. After adjustment for educational attainment, the percentage gainfully employed was 11-12% less among subjects having received benefits in childhood. This negative association between having received a benefit in childhood and gainful employment in adulthood was restricted to the low educational group. INTERPRETATION: Persons with health problems in childhood did not achieve the education, employment and income they should have been capable of judged by their general ability and the education they had actually achieved. PMID- 16477280 TI - [Generic drugs instead of brand drugs prescriptions--long overdue]. AB - BACKGROUND: Generic substitution of drugs was introduced in Norway in 2001. This implies an obligation for pharmacies to substitute the prescribed drug with a generic equivalent. Instead of using the drug manufacturers' trade names, doctors can prescribe drugs by using the generic (non-proprietary) names. The present paper gives an overview over current practice in different countries and discusses advantages and drawbacks. MATERIALS AND METHODS: Literature on generic substitution and generic prescription was sought on PubMed. Regulatory authorities in Norway and other countries were approached for additional information. RESULTS: In the United Kingdom, generic prescription has been dominating for many years. In Sweden, a successful trial period with generic prescribing was recently reported. We have not identified any pharmacological arguments against generic prescription. INTERPRETATION: Available data suggest that generic prescribing of drugs should be encouraged. Generic prescribing is likely to improve patient safety by assuring that the prescribed drug is in fact what is dispensed. PMID- 16477281 TI - [Monitoring in acute cerebral infarction]. AB - BACKGROUND: Cerebral ischaemia is potentially reversible during the first few hours. Monitoring of physiological variables such as temperature, blood pressure, heart rhythm, oxygen saturation and cerebral blood flow may be important for efficient treatment and reduction of the infarct volume. MATERIAL AND METHODS: This article is based on a review of relevant articles in international journals over the last 15 years on the pathophysiology of acute ischaemic stroke and treatment in intensive stroke units compared with conventional stroke units. RESULTS AND DISCUSSION: Two pilot studies indicate that continuous monitoring in intensive stroke units improves outcome compared with conventional stroke units. Future studies must draw the balance between continuous monitoring and mobilization in the early phase of ischaemic stroke. PMID- 16477283 TI - [Experiences with botulinum toxin injections against spasticity in children]. AB - BACKGROUND: Botulinum toxin injection is one of the newer options in the treatment of spasticity. Treatment with botulinum toxin is always combined with physiotherapy and often with casting. We show the extent to which botulinum toxin treatment has been taken into use in our department and discuss advantages and disadvantages of giving botulinum toxin injections at local hospitals. MATERIAL AND METHODS: 88 children with cerebral palsy aged 14 months to 16 years were treated with botulinum toxin between March 2000 and Dec. 2004. Injections were given in an outpatient setting, usually with the patient sedated with midazolam. Clinical examination after injection included assessment of spasticity and range of movement of joints. Motor function was videotaped. Side effects were continuously supervised. RESULTS: We performed 278 treatments; during the last year (2004) 7 treatments a month on average. 59 children were injected in lower limbs only, 14 were injected in upper limbs only, and 15 were injected in both upper and lower limbs. No serious side effects were recorded, neither from the botulinum toxin itself nor related to the injection procedure. INTERPRETATION: Assessment of indications for the use of botulinum toxin is now part of the medical follow up for children with spastic cerebral palsy. Treatment can safely be given at the paediatrics department in a local hospital. Injection in superficial muscles of the lower limbs is an easy task, while injection in small muscles and deep-seated muscles requires more special skills. PMID- 16477284 TI - [Risk factors for accidental injuries in the construction industry]. AB - BACKGROUND: The construction industry is prone to accidents. In Norway there is little research-based knowledge on accident pattern and risk factors. The main objective of the study was to examine whether in-depth studies of accidents in this industry, leading to injuries registered by the health services, could identify preventable risk factors and preventive measures. MATERIAL AND METHODS: 50 accidents in the construction industry were thoroughly investigated. All had led to serious personal injury. Patients were interviewed after treatment in emergency wards and hospitals in Oslo, and inspections of the accident sites were performed. An interdisciplinary expert group discussed the accidents in order to identify risk factors and suggest preventive measures. The accident reports in the study were compared to reports from the Labour Inspectorate on the same accidents. RESULTS: The investigation identified several risk factors, and a number of detailed preventive measures were proposed. We submitted these proposals to the authorities and organisations involved. Accidents because of time pressure and electricity were two main groups. More than one third of the respondents said that time pressure had contributed to the accident. Accident risk caused by time pressure can be reduced by avoiding piecework contracts, unrealistic time limits, and the use of day penalties for breach of contract. Possible preventive measures for electric injuries imply modification of the reporting system and of the work organisation so that workers can comply with the regulations more easily. The study reports generally contained more information of relevance for prevention than the routine reports from the Labour Inspectorate did. For accidents reported by the Inspectorate to the police, their reports were more informative than the study reports. INTERPRETATION: By the method presented, we obtained detailed information about accidents and preventable risk factors, so that preventive measures could be proposed. The method can also be used for other types of accidents. PMID- 16477282 TI - [Protective silicone prosthesis prior to radiotherapy of recurrent colonic cancer]. AB - Radiation therapy for abdominal recurrence of colon cancer is rarely an option due to subsequent bowel injury. Our case is a woman who underwent resection for a large retroperitoneal recurrence of caecal cancer. Tumour deposits encasing the iliac vessels had to be left behind. A silicone breast prosthesis for displacement of the abdominal content was implanted, allowing postoperative irradiation with 50 Gy. The prosthesis was removed once radiotherapy was accomplished; tumour regression was then complete. Complications are described, so are indications for surgical management of local recurrences of colonic origin as well as technical aspects of abdominal implantation of displacing prostheses. At follow-up after eighteen months the patient has no signs of enteropathy, she enjoys a good quality of life, and she is free of disease. Still, her prognosis is considered uncertain. PMID- 16477285 TI - [Atomoxetine in the treatment of ADHD]. PMID- 16477286 TI - [Approaching patients who require palliative care]. AB - The aim of palliative medicine is the best possible quality of life. Based upon literature and clinical experience we address factors of importance for the meeting between the palliative patient and the physician. Family and network, personality and behaviour vary between palliative patients and have effect upon their coping; these factors should be reflected in the meeting between the physician and the patient. Communication with a palliative patient also aims at a systematic assessment of his or her various symptoms and the physician should have a broad armamentarium of communications skills. The optimal way to assess symptoms is by use of clinical interviews supplemented with standardized measurements. Instruments such as the Edmonton Symptom Assessment System are very useful in the assessment of the commonest symptoms. The assessments should include patients' perspectives and what priority they give to relevant interventions. Treatment should be evaluated systematically in order to avoid ineffective treatments and to reduce side effects and interactions. The aim is to give patients as good and as long a time as possible in the place in which they want to spend the last part of their lives. Palliative medicine often combines the art of medicine with new technology. The focus on quality of life and the patient perspective is paramount, and the approach to the patient should reflect this. PMID- 16477287 TI - [The dying patient]. AB - All doctors in clinical practice have to face dying patients. In order to give help to the patient and his nearest family, the doctor needs knowledge about the dying process and how to relieve suffering. Based on relevant literature and own clinical experience, we discuss the challenges of identifying the terminal phase, ethical issues concerning medical treatment, and how to offer adequate symptom relief. We describe how to relieve pain and other symptoms, and how to organise the care of the dying. Drugs that may be useful in relieving suffering are described with dosing proposals. Family members have to be helped as well with information and reassurance. A Norwegian Standard for Palliation focusing on the organisation of palliative and terminal care has recently been published. PMID- 16477288 TI - [Palliative sedation to the dying]. AB - BACKGROUND: Guidelines for palliative sedation to dying patients were issued by the Council for Medical Ethics of the Norwegian Medical Association in 2001. This study is a follow up of these guidelines. MATERIAL AND METHODS: In 2002 an anonymous questionnaire was sent to 400 hospital departments. After two years, only 12 questionnaires had been returned. In addition to this survey we present data from a survey among a random sample of 1539 doctors. RESULTS: Ten of the 12 treatments which were registered in detail concerned patients with advanced cancer and in great discomfort. Average age was 49. Pain, nausea, convulsions and dyspnoea were the most common symptoms. Nine out of the 12 patients were in pain. In no case the sedation was assessed to have been avoidable with more health care resources. Relatively few patients had been examined psychologically. The decisions were taken by several caregivers, always in accordance with the family's wish. Six of the patients whose conscience level was raised were sedated again. Midazolam and morphine were the two mostly-used drugs; doses used varied considerably. 12% of the surveyed physicians said that one or more dying patients had been given palliative sedation in their department during the last 12 months. INTERPRETATION: Palliative sedation for dying patients is probably used infrequently in Norway, and not to hasten death. In the cases which we have registered, palliative sedation was given to patients in great discomfort. PMID- 16477289 TI - [Female circumcision--complications and treatment]. AB - The majority of women with extensive forms of genital cutting develop one or more chronic complications such as dysmenorrhea, dyspareunia, pain and cysts in the perineum, vaginal obstruction with haematocolpos, relative urine retention and recurrent urinary tract infections. Extensive forms of circumcision also influence childbirths. The severity of the cutting is associated with the probability of developing later complications. The women's clinics at the regional hospital in Norway have established outpatient clinics to receive women with complications after genital cutting. The aim was to develop an adequate health service to the affected. In order to improve the access to care and to ensure anonymity the women may refer themselves. During 2004, a total of 60 women were treated. The majority suffered from poor urinary flow, pain at micturition, dysmenorrhea and dyspareunia. Reconstruction of the vaginal orifice was performed to relieve some of the discomforts. The numbers of women who visit the clinics are increasing. The surgical procedure itself is not technically difficult, but the consultation before and after the surgery require cultural sensitivity. As health care personnel we can influence the affected to realise that genital cutting is an assault against a small girl. Norwegian health care workers need to learn more about how to communicate well about the medical as well as the cultural and mental aspects of genital cutting. PMID- 16477290 TI - [Research with radioactive isotopes using humans in Norway]. PMID- 16477292 TI - [Anatomic trauma scoring following accidents]. PMID- 16477294 TI - [CYP genotyping in psychopharmacological treatment]. PMID- 16477295 TI - [Atomoxetine--an addition]. PMID- 16477297 TI - [Alcohol-injured statistics?]. PMID- 16477299 TI - [Are children getting the right drugs?]. PMID- 16477300 TI - [Are children getting dangerous anesthetics?]. PMID- 16477301 TI - [Medical terminology education]. PMID- 16477302 TI - [Is a patient per definition resource-weak]. PMID- 16477303 TI - ["Osteoporosis" and physiological variation]. PMID- 16477309 TI - Two key residues in ephrinB3 are critical for its use as an alternative receptor for Nipah virus. AB - EphrinB2 was recently discovered as a functional receptor for Nipah virus (NiV), a lethal emerging paramyxovirus. Ephrins constitute a class of homologous ligands for the Eph class of receptor tyrosine kinases and exhibit overlapping expression patterns. Thus, we examined whether other ephrins might serve as alternative receptors for NiV. Here, we show that of all known ephrins (ephrinA1-A5 and ephrinB1-B3), only the soluble Fc-fusion proteins of ephrinB3, in addition to ephrinB2, bound to soluble NiV attachment protein G (NiV-G). Soluble NiV-G bound to cell surface ephrinB3 and B2 with subnanomolar affinities (Kd = 0.58 nM and 0.06 nM for ephrinB3 and B2, respectively). Surface plasmon resonance analysis indicated that the relatively lower affinity of NiV-G for ephrinB3 was largely due to a faster off-rate (K(off) = 1.94 x 10(-3) s(-1) versus 1.06 x 10(-4) s(-1) for ephrinB3 and B2, respectively). EphrinB3 was sufficient to allow for viral entry of both pseudotype and live NiV. Soluble ephrinB2 and B3 were able to compete for NiV-envelope-mediated viral entry on both ephrinB2- and B3-expressing cells, suggesting that NiV-G interacts with both ephrinB2 and B3 via an overlapping site. Mutational analysis indicated that the Leu-Trp residues in the solvent exposed G-H loop of ephrinB2 and B3 were critical determinants of NiV binding and entry. Indeed, replacement of the Tyr-Met residues in the homologous positions in ephrinB1 with Leu-Trp conferred NiV receptor activity to ephrinB1. Thus, ephrinB3 is a bona fide alternate receptor for NiV entry, and two residues in the G-H loop of the ephrin B-class ligands are critical determinants of NiV receptor activity. PMID- 16477310 TI - trt-1 is the Caenorhabditis elegans catalytic subunit of telomerase. AB - Mutants of trt-1, the Caenorhabditis elegans telomerase reverse transcriptase, reproduce normally for several generations but eventually become sterile as a consequence of telomere erosion and end-to-end chromosome fusions. Telomere erosion and uncapping do not cause an increase in apoptosis in the germlines of trt-1 mutants. Instead, late-generation trt-1 mutants display chromosome segregation defects that are likely to be the direct cause of sterility. trt-1 functions in the same telomere replication pathway as mrt-2, a component of the Rad9/Rad1/Hus1 (9-1-1) proliferating cell nuclear antigen-like sliding clamp. Thus, the 9-1-1 complex may be required for telomerase to act at chromosome ends in C. elegans. Although telomere erosion limits replicative life span in human somatic cells, neither trt-1 nor telomere shortening affects postmitotic aging in C. elegans. These findings illustrate effects of telomere dysfunction in C. elegans mutants lacking the catalytic subunit of telomerase, trt-1. PMID- 16477311 TI - Seriation in paleontological data using markov chain Monte Carlo methods. AB - Given a collection of fossil sites with data about the taxa that occur in each site, the task in biochronology is to find good estimates for the ages or ordering of sites. We describe a full probabilistic model for fossil data. The parameters of the model are natural: the ordering of the sites, the origination and extinction times for each taxon, and the probabilities of different types of errors. We show that the posterior distributions of these parameters can be estimated reliably by using Markov chain Monte Carlo techniques. The posterior distributions of the model parameters can be used to answer many different questions about the data, including seriation (finding the best ordering of the sites) and outlier detection. We demonstrate the usefulness of the model and estimation method on synthetic data and on real data on large late Cenozoic mammals. As an example, for the sites with large number of occurrences of common genera, our methods give orderings, whose correlation with geochronologic ages is 0.95. PMID- 16477314 TI - Excitable but lacking in energy: contradictions in the human epileptic hippocampus. PMID- 16477313 TI - Posttraumatic epilepsy: a major problem in desperate need of major advances. AB - This brief review is meant to provide an update on the data from clinical and laboratory studies that have provided insight into the mechanisms underlying the development of epilepsy following traumatic brain injury (TBI). The link between severe brain trauma and epilepsy in humans is well recognized. However, we have yet to identify an effective intervention to prevent the development of epilepsy in patients who are at risk after TBI. Laboratory studies, which have relied primarily on the fluid-percussion model, have documented long-term hyperexcitability associated with TBI, and recent studies are shedding light on the structural and electrophysiological abnormalities that may underlie epileptogenesis in this setting. Nonetheless, given the extent of the clinical problem and our current state of knowledge, this area of epilepsy research deserves far more attention. PMID- 16477312 TI - Permissive transcriptional activity at the centromere through pockets of DNA hypomethylation. AB - DNA methylation is a hallmark of transcriptional silencing, yet transcription has been reported at the centromere. To address this apparent paradox, we employed a fully sequence-defined ectopic human centromere (or neocentromere) to investigate the relationship between DNA methylation and transcription. We used sodium bisulfite PCR and sequencing to determine the methylation status of 2,041 CpG dinucleotides distributed across a 6.76-Mbp chromosomal region containing a neocentromere. These CpG dinucleotides were associated with conventional and nonconventional CpG islands. We found an overall hypermethylation of the neocentric DNA at nonconventional CpG islands that we designated as CpG islets and CpG orphans. The observed hypermethylation was consistent with the presence of a presumed transcriptionally silent chromatin state at the neocentromere. Within this neocentric chromatin, specific sites of active transcription and the centromeric chromatin boundary are defined by DNA hypomethylation. Our data demonstrate, for the first time to our knowledge, a correlation between DNA methylation and centromere formation in mammals, and that transcription and "chromatin-boundary activity" are permissible at the centromere through the selective hypomethylation of pockets of sequences without compromising the overall silent chromatin state and function of the centromere. PMID- 16477315 TI - The hyperinhibition hypothesis in epileptogenesis: an assessment of the evidence. PMID- 16477316 TI - The toll receptor family: from microbial recognition to seizures. PMID- 16477317 TI - Dancing the delta shuffle: neurosteroids regulate GABAA receptor expression. PMID- 16477318 TI - Endocannabinoids: a critical regulator of activity in the developing brain. PMID- 16477319 TI - Struggling with Rasmussen's syndrome. PMID- 16477320 TI - Is breast milk the best for babies of mothers on levetiracetam? PMID- 16477321 TI - Impact of epilepsy surgery on developing minds: how do we weigh the consequences? PMID- 16477322 TI - In the end, new antiepileptics may be less expensive than older antiepileptics for geriatric patients. PMID- 16477323 TI - What does flumazenil PET add to an evaluation for temporal lobectomy? PMID- 16477324 TI - PhyloGibbs: a Gibbs sampling motif finder that incorporates phylogeny. AB - A central problem in the bioinformatics of gene regulation is to find the binding sites for regulatory proteins. One of the most promising approaches toward identifying these short and fuzzy sequence patterns is the comparative analysis of orthologous intergenic regions of related species. This analysis is complicated by various factors. First, one needs to take the phylogenetic relationship between the species into account in order to distinguish conservation that is due to the occurrence of functional sites from spurious conservation that is due to evolutionary proximity. Second, one has to deal with the complexities of multiple alignments of orthologous intergenic regions, and one has to consider the possibility that functional sites may occur outside of conserved segments. Here we present a new motif sampling algorithm, PhyloGibbs, that runs on arbitrary collections of multiple local sequence alignments of orthologous sequences. The algorithm searches over all ways in which an arbitrary number of binding sites for an arbitrary number of transcription factors (TFs) can be assigned to the multiple sequence alignments. These binding site configurations are scored by a Bayesian probabilistic model that treats aligned sequences by a model for the evolution of binding sites and "background" intergenic DNA. This model takes the phylogenetic relationship between the species in the alignment explicitly into account. The algorithm uses simulated annealing and Monte Carlo Markov-chain sampling to rigorously assign posterior probabilities to all the binding sites that it reports. In tests on synthetic data and real data from five Saccharomyces species our algorithm performs significantly better than four other motif-finding algorithms, including algorithms that also take phylogeny into account. Our results also show that, in contrast to the other algorithms, PhyloGibbs can make realistic estimates of the reliability of its predictions. Our tests suggest that, running on the five species multiple alignment of a single gene's upstream region, PhyloGibbs on average recovers over 50% of all binding sites in S. cerevisiae at a specificity of about 50%, and 33% of all binding sites at a specificity of about 85%. We also tested PhyloGibbs on collections of multiple alignments of intergenic regions that were recently annotated, based on ChIP-on-chip data, to contain binding sites for the same TF. We compared PhyloGibbs's results with the previous analysis of these data using six other motif-finding algorithms. For 16 of 21 TFs for which all other motif-finding methods failed to find a significant motif, PhyloGibbs did recover a motif that matches the literature consensus. In 11 cases where there was disagreement in the results we compiled lists of known target genes from the literature, and found that running PhyloGibbs on their regulatory regions yielded a binding motif matching the literature consensus in all but one of the cases. Interestingly, these literature gene lists had little overlap with the targets annotated based on the ChIP-on-chip data. The PhyloGibbs code can be downloaded from http://www.biozentrum.unibas.ch/~nimwegen/cgi-bin/phylogibbs.cgi or http://www.imsc.res.in/~rsidd/phylogibbs. The full set of predicted sites from our tests on yeast are available at http://www.swissregulon.unibas.ch. PMID- 16477325 TI - Functional alignment of regulatory networks: a study of temperate phages. AB - The relationship between the design and functionality of molecular networks is now a key issue in biology. Comparison of regulatory networks performing similar tasks can provide insights into how network architecture is constrained by the functions it directs. Here, we discuss methods of network comparison based on network architecture and signaling logic. Introducing local and global signaling scores for the difference between two networks, we quantify similarities between evolutionarily closely and distantly related bacteriophages. Despite the large evolutionary separation between phage lambda and 186, their networks are found to be similar when difference is measured in terms of global signaling. We finally discuss how network alignment can be used to pinpoint protein similarities viewed from the network perspective. PMID- 16477327 TI - Serotonergic mechanisms: a potential link between affective disorders and cardiovascular risk. AB - Cardiovascular disease and major depression are highly prevalent disorders in our society. Evidence has been found that confirms a reciprocal relationship between mechanisms of depression and those of cardiovascular pathology. This possible feedback between both pathologies is a subject of great concern. In recent years some studies suggest that platelets and serotonergic mechanisms could be involved in both conditions. The present review seeks a better understanding of the mechanisms that could link depression with an enhanced cardiovascular risk. PMID- 16477328 TI - Inflammation, the key to much pathology. Highlights from the 7th World Congress on Inflammation, held August 20-24, 2005, in Melbourne, Australia. AB - From the World Congress on Inflammation, held August 20-24, 2005 in Melbourne, Australia, new targets and new drugs for inflammation of the respiratory system (asthma, allergic rhinitis, chronic obstructive pulmonary disease and cystic fibrosis), inflammatory bowel disease (ulcerative colitis and Crohn's disease), arthritis (rheumatoid and osteoarthritis), atherosclerosis and cancer are discussed. PMID- 16477329 TI - Cardiac regeneration and repair: do they really exist? PMID- 16477330 TI - Association of p53 codon 72 polymorphism with risk of hypopharyngeal squamous cell carcinoma in Taiwan. AB - BACKGROUND: p53 polymorphism at codon 72 is a known risk marker for various malignancies, but it has not been studied in hypopharyngeal cancer. This study investigated the genotype distribution of p53 codon 72 polymorphism in hypopharyngeal cancer patients and non-cancer controls matched for age, gender, alcohol consumption and smoking habit. METHODS: Genomic DNA was extracted from peripheral blood cells of 53 patients with hypopharyngeal cancer and 53 non cancer controls. Codon 72 polymorphism of p53 was identified by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: Patients with hypopharyngeal cancer had higher frequencies of Pro/Pro (26.4% vs. 13.2%) and Pro/Arg (51.0% vs. 45.3%) but lower frequencies of Arg/Arg (22.6% vs. 45.1%) compared to controls. Compared to Arg/Arg genotypes, Pro/Pro genotypes had a relative risk of hypopharyngeal cancer of 3.667 (95% confidence interval, 1.16 11.56; p = 0.03). As a group, patients with Pro/Pro or Arg/Pro who were carriers of the Pro allele had a higher relative risk of hypopharyngeal cancer compared to Arg homozygous carriers (odds ratio, 2.415; 95% confidence interval, 1.04-5.64; p = 0.04). CONCLUSION: This study demonstrated that p53 codon 72 Pro homozygosity is associated with a higher risk of developing hypopharyngeal cancer. PMID- 16477331 TI - Biliary cysts in children--long-term follow-up in Taiwan. AB - BACKGROUND: This study analyzed the clinical presentation, postoperative morbidity and mortality and incidence of associated extrahepatic biliary atresia in children with biliary cysts in Taiwan. METHODS: We retrospectively reviewed the records of 158 pediatric patients with biliary cysts seen between June 1981 and July 2004, with follow-up ranging from 12 months to 22 years (mean, 11.2 +/- 6.1 years). Patients were divided into three groups: biliary atresia-associated biliary cyst (BABC, 21 patients), non-biliary atresia-associated choledochal cyst (NBACC) in infancy (37 patients), and late NBACC (> 1 year of age, 100 patients). RESULTS: BABC accounted for 36.2% of the infantile biliary cysts in this study. Extrahepatic cysts in late NBACC had a greater mean diameter than those in infantile NBACC and BABC (21.5 mm vs. 16.0 mm vs. 7.9 mm, p < 0.001). Cholangitis was the most serious complication within 3 months postoperatively in all three groups, resulting in four deaths (two in the infantile NBACC group and one each in the other two groups). Liver cirrhosis developed during long-term follow-up in nine of the 21 patients with BABC, four of whom died. Three of these nine patients underwent liver transplantation and remained well during follow-up. Chronic complications in NBACC occurred mainly in late IVa cases, with persistent intrahepatic dilatation developing in 12 of 35 patients and intrahepatic stones in five. Elevation of serum alanine aminotransferase (ALT) was found preoperatively in 85% of late NBACC and 35% of infantile NBACC cases. Postoperative normalization of ALT occurred after a mean of 152 +/- 23 days and 158 +/- 67 days in late NBACC and infantile NBACC, respectively. Higher ALT levels before operation were associated with a longer period until normalization. CONCLUSION: The possibility of BABC must be included in the differential diagnosis when a small extrahepatic cyst (< 8 mm in diameter) with prolonged jaundice is found in infancy. Postoperative follow-up is essential for patients with NBACC due to their frequently prolonged elevation of serum ALT and possibility of residual intrahepatic dilatation. Cholangitis was the major cause of death within 3 months postoperatively in this study. PMID- 16477332 TI - Magnetic resonance imaging of seminal vesicle cyst associated with ipsilateral urinary anomalies. AB - BACKGROUND: Seminal vesicle cysts rarely cause symptoms. Data on long-term follow up from childhood to adulthood are lacking. The study analyzed the magnetic resonance imaging (MRI) and follow-up results of this condition. METHODS: From 1991 to 1996, seminal vesicle cyst was diagnosed in 13 boys (mean age, 12 years; range, 7-15 years), six of whom had long-term follow-up data. The clinical symptoms and MRI findings at diagnosis and at follow-up were analyzed. RESULTS: The seminal vesicle cyst was on the right side in six patients and on the left in seven. The size varied, ranging from 1.0 x 1.3 x 1.4 to 4.4 x 3.1 x 3.6 cm. All showed high signal intensity on T2-weighted images but variable signal intensity on T1-weighted images. Associated urinary tract anomalies included renal anomalies (dysplasia in 2 patients, agenesis in 11), ectopic ureteral orifice (11), hydroureter (6), and vertebral anomalies (2). One of the six patients with follow-up had repeated urinary tract infection and underwent surgical resection of the cyst 8 years after the diagnosis. The other five had no symptoms during the follow-up period. Three of the six patients had repeat MRI after a median of 11 years, which showed slight cyst enlargement and increased T1-weighted signal intensity. CONCLUSION: Most seminal vesicle cysts were asymptomatic and did not change during long-term follow-up. MRI is a powerful tool for detecting seminal vesicle cysts and in delineating associated congenital anomalies of the urogenital tract. PMID- 16477333 TI - Imaging factors related to rotator cuff tear in patients with deltoid contracture. AB - BACKGROUND: Bony deformity and muscular malfunction around the shoulder induced by deltoid contracture may influence rotator cuff function and lead to subsequent tearing. The purpose of this study was to investigate the imaging and clinical factors related to rotator cuff tear in patients with deltoid contracture. METHODS: We retrospectively reviewed 48 shoulders in 44 patients with magnetic resonance imaging diagnosis of deltoid contracture and surgically-proven rotator cuff tear. These shoulders were subdivided into partial-thickness and full thickness tear groups. Another 17 shoulders in 17 patients with deltoid contracture but no evident rotator cuff tear were included as the control group. The characteristics, including age, gender and imaging features, of these three groups were compared. RESULTS: Two patients who underwent initial operations for cuff tears required secondary operations for distal release of deltoid contracture. The age of patients with full-thickness tear was significantly greater than that of patients with partial-thickness tear or without tear. The winging angle of the glenoid (WAG) in the full-thickness tear group was significantly greater than that in the partial-thickness tear group. Patients with partial-thickness tear also had greater WAG than those without tear. However, WAG was not significantly correlated with the fibrotic size or age of the patient. CONCLUSION: In patients with deltoid contracture, age and WAG are associated with the development of rotator cuff tear. To prevent the occurrence of rotator cuff tear, early surgical release of deltoid contracture is highly recommended, especially in patients with greater WAG. PMID- 16477334 TI - Comparison of the Gow-Gates mandibular block and inferior alveolar nerve block using a standardized protocol. AB - BACKGROUND: Although several previous studies have compared the efficacy of Gow Gates mandibular block (GGMB) and inferior alveolar nerve block (IANB), the results remain controversial. This study used an objective, standardized and precise protocol to evaluate and compare the effectiveness and success rate of GGMB and IANB. METHODS: The study group consisted of 162 patients (93 males and 69 females) who were randomly allocated to receive GGMB or IANB for extraction of third molars. Both methods used 2.7 mL of 2% xylocaine for each patient. Pulpal and gingival tissue anesthesia of mandibular central incisors, canines, first premolars and first molars were evaluated at 0, 5, 10, 15 and 60 minutes after injection of local anesthetic solution using both an electric pulp tester and a sharp explorer. RESULTS: The success rates of pulpal anesthesia in the IANB group (central incisor, 6%; canine, 37%; first premolar, 54%; first molar, 88%) were not significantly different from the GGMB group (central incisor, 8.1%; canine, 37.1%; first premolar, 54.8%; first molar, 83.9%). All subjects achieved 100% lip numbness with both methods. At 60 minutes after injection, the success rates of gingival tissue anesthesia in canine buccal and lingual areas were higher in the IANB group (100% and 100%, respectively) than in the GGMB group (91.9% and 93.5%, respectively). In the molar buccal area, the success rates at 5 and 60 minutes after injection were higher in the IANB group (97% and 100%, respectively) than in the GGMB group (88.7% and 91.9%, respectively). Furthermore, the success rates in the molar lingual area at 10, 15 and 60 minutes after injection were higher in the IANB group (100%, 100% and 100%, respectively) than in the GGMB group (91.9%, 93.5% and 91.9%, respectively). Although IANB achieved higher success rates of gingival tissue anesthesia in some gingival areas, no significant difference between the two methods was found in overall efficacy. CONCLUSION: This study demonstrated that the efficacy of pulpal and gingival tissue anesthesia are not significantly different between the GGMB and IANB methods. PMID- 16477335 TI - Thin-plate spline analysis of the effects of face mask treatment in children with maxillary retrognathism. AB - BACKGROUND: Face mask therapy is indicated for growing patients who suffer from maxillary retrognathia. Most previous studies used conventional cephalometric analysis to evaluate the effects of face mask treatment. Cephalometric analysis has been shown to be insufficient for complex craniofacial configurations. The purpose of this study was to investigate changes in the craniofacial structure of children with maxillary retrognathism following face mask treatment by means of thin-plate spline analysis. METHODS: Thirty children with skeletal Class III malocclusions who had been treated with face masks were compared with a group of 30 untreated gender-matched, age-matched, observation period-matched, and craniofacial configuration-matched subjects. Average geometries, scaled to an equivalent size, were generated by means of Procrustes analysis. Thin-plate spline analysis was then performed for localization of the shape changes. RESULTS: Face mask treatment induced a forward displacement of the maxilla, a counterclockwise rotation of the palatal plane, a horizontal compression of the anterior border of the symphysis and the condylar region, and a downward deformation of the menton. The cranial base exhibited a counterclockwise deformation as a whole. CONCLUSION: We conclude that thin-plate spline analysis is a valuable supplement to conventional cephalometric analysis. PMID- 16477336 TI - Evolution of human herpesvirus type 8-associated gastric Kaposi's sarcoma following corticosteroid treatment for asthma. AB - The association of gastric Kaposi's sarcoma (KS) with human herpesvirus type 8 (HHV-8) may be found in immunosuppressed patients such as those with AIDS or transplant recipients. A 64-year-old man with a 2-year history of corticosteroid treatment was admitted due to the impression of chronic obstructive pulmonary disease with secondary infection. Abdominal fullness and tarry stool led to the performance of panendoscopy, which revealed two hypertrophic gastric mucosal lesions. These lesions had transformed into a large polyp 2 months later. KS was not diagnosed until a third endoscopic biopsy was performed. The polyp further transformed into a flattened, elongated tumor within a month. The patient died from acute cardiopulmonary insufficiency 4 days after radical gastrectomy. Polymerase chain reaction study identified HHV-8 in all biopsied specimens and resected tumor lesions. Immunostaining further demonstrated the virus in the tumor cells. Both of these methods seemed more sensitive in diagnosing KS than histologic examination of small biopsied specimens. This case suggests the existence of a relationship between gastric KS and HHV-8 infection. PMID- 16477337 TI - Primary intrathoracic goiter. AB - Nodular goiter is a common disease in Taiwan, and substernal or intrathoracic goiters are not infrequent. However, intrathoracic goiters are mainly of the secondary type and primary intrathoracic goiters are rarely seen. We report a 55 year-old woman who was incidentally found to have an ectopic goiter located in the anterior upper mediastinum with the initial presenting symptom of productive cough. Imaging studies including chest X-ray and computed tomography identified the lesion, and 131I-uptake scan showed weak uptake in the thorax. Surgical removal via thoracotomy was performed and the diagnosis was confirmed by pathology. A primary intrathoracic goiter, although rare, should also be considered in the differential diagnosis of mediastinal tumor. PMID- 16477338 TI - Primary amyloidosis of the urinary bladder. AB - Amyloidosis is a systemic disease that usually occurs in the gastrointestinal tract or in muscular or adipose tissue. Primary amyloidosis of the urinary bladder is a rare disease that can mimic bladder cancer on cystoscopic examination as well as in its clinical presentation of painless gross hematuria. This report describes a 49-year-old male with repeated painless gross hematuria, who underwent transurethral resection of a suspected bladder tumor. Pathologic examination revealed papillary urothelial hyperplasia with vascular ectasia and no signs of malignancy. Massive gross hematuria occurred 2.5 years later. Cystoscopy showed multiple papillary lesions with yellowish-brown submucosal plaques on the posterior bladder wall. A second transurethral tumor resection was performed and histologic examination revealed plasma cell infiltration and eosinophilic amorphous deposits in the subepithelial stroma and vascular wall. The deposits were positive for Congo red and apple-green birefringence under polarized light examination but negative for Masson's trichrome stain, indicating that they were not fibrotic in nature. Hence, the diagnosis of amyloidosis of the urinary bladder was confirmed. Screening for amyloidosis was negative in other organ systems and the patient has remained disease-free up to the last follow-up 4 years after the second transurethral resection. Amyloidosis should be considered in the differential diagnosis of patients with recurrent hematuria who have symptoms characteristic of bladder cancer but negative pathologic study for malignancy. Correct diagnosis relies on clinical alertness and the use of a special staining technique during pathologic examination. PMID- 16477339 TI - T-fastener gastropexy and percutaneous gastrostomy under CT-fluoroscopic guidance in a patient with partial gastrectomy. AB - The real-time images of computed tomography (CT)-fluoroscopy provide an excellent means of guidance for percutaneous interventions. We describe the performance of T-fastener gastropexy and percutaneous gastrostomy under CT-fluoroscopic guidance in a 59-year-old woman who had received total pharyngolaryngectomy for hypopharyngeal cancer and partial gastrectomy with Billroth II anastomosis for bleeding gastric ulcer 10 years before this operation. The previous gastric operation altered the gastrointestinal anatomy and made conventional fluoroscopic guided percutaneous gastrostomy extremely difficult and risky. The T-fastener gastropexy and percutaneous gastrostomy were accomplished smoothly in a single session using CT-fluoroscopic guidance. This modified method of percutaneous gastrostomy may be useful in patients with anatomic distortion due to previous gastric surgery. PMID- 16477340 TI - Neuropsychiatric disturbances and hypopituitarism after traumatic brain injury in an elderly man. AB - Neuropsychiatric or cognitive disturbances are common complications after traumatic brain injury. They are commonly regarded as irreversible sequelae of organic brain injuries. We report a case of hypopituitarism in a 77-year-old man who presented with long-term neuropsychiatric disturbances, including cognitive impairment, disturbed sleep patterns, personality change, loss of affect, and visual and auditory hallucinations after a traumatic subdural hemorrhage. The treatment response to hormone replacement therapy was nearly complete. Hypopituitarism is rarely considered in patients who sustain traumatic brain injury and the neuropsychiatric manifestations of posttraumatic hypopituitarism have rarely been reported. This case highlights the importance of hypopituitarism as a potential reversible cause of neuropsychiatric disturbances after traumatic brain injury. PMID- 16477341 TI - Seventeen alpha-hydroxylase deficiency. AB - Seventeen alpha-hydroxylase deficiency (17OHD) is a rare form of congenital adrenal hyperplasia in which defects in the biosynthesis of cortisol and sex steroid result in mineralocorticoid excess, hypokalemic hypertension and sexual abnormalities such as pseudohermaphroditism in males, and sexual infantilism in females. The disease is inherited in an autosomal recessive pattern, and is caused by mutations in the gene encoding cytochrome P450c17 (CYP17), which is the single polypeptide that mediates both 17alpha-hydroxylase and 17,20-lyase activities. We report the case of a 15-year-old patient with 17OHD who had a female phenotype but male karyotype (46,XY). The diagnosis was made based on classical clinical features, biochemical data and molecular genetic study. Two mutations were identified by polymerase chain reaction amplification and sequencing, including a S106P point mutation in exon 2 and a 9-bp (GACTCTTTC) deletion from nucleotide position 1519 in exon 8 of CYP17. The first of these mutations was found in the father and the second in the mother, and both have been previously reported in Asia. The patient's hypertension and hypokalemia resolved after glucocorticoid replacement and treatment with potassium-sparing diuretics. Sex hormone replacement was prescribed for induction of sexual development and reduction of the final height. Prophylactic gonadectomy was scheduled. In summary, 17OHD should be suspected in patients with hypokalemic hypertension and lack of secondary sexual development so that appropriate therapy can be implemented. PMID- 16477343 TI - [The use of transgenic animals in biomedical research in Germany. Part 2: Ethical evaluation of the use of transgenic animals in biomedical research and perspectives for the changeover in research to research animal-free methods]. AB - As a rule, transgenic animals are being used in in vivo experiments to examine gene functions, their regulation or the contribution of genetic alterations to the development of diseases. Many transgenic animals already are affected in their wellbeing due to the genetic modification alone regardless of the procedures performed with them. Moreover, it is to be questioned whether the experimental use of transgenic animals led to results that were of such outstanding scientific relevance that they legitimated the suffering of the animals. In order to point to possible approaches to avoiding the use of transgenic animals in the areas of research identified, subsequent investigations aimed at collecting information on non-animal test methods that might be applied in pursuing the aforesaid questions. In particular, these were non-animal test methods that make use of genetic techniques. Amongst these are in vitro cell culture methods with genetically modified cells, such as the so called Transfected Cell Array, as well as in vitro test methods, in which specifically targeted genes can be turned on or off selectively for example by the so-called RNA interference technique or by antisense oligonucleotide genes. Since such technologies can also be applied to cell cultures with human cells, investigations with these methods enable direct information on the function of human genes. Even though a one to one replacement of animal experiments with transgenic animals by non-animal test methods is considered unlikely, from the point of view of animal welfare the broad spectrum of already available non animal test methods with which to study the function of genes and genetically caused pathophysiological reactions proves that waiving of animal tests with transgenic animals is possible without impeding biomedical research. Even if it cannot be totally excluded that some very specific questions linked to the respective animal experiment might not be pursued for the time being, nevertheless research that would be restricted to modern and ethically acceptable in vitro test methods would certainly conceive its very own questions to pursue and solve the problems currently faced by biomedical research. It is against this background that it is to be welcomed that the German Federal Government currently actively promotes the further development of genetechnological non-animal test methods. In order to ensure that these funding measures will make an effective contribution to reducing animal experiments, as spelled out by the government itself, the conversion of genetechnological research, just like biomedical research as a whole, to non-animal testing methods should be supported by concrete political actions. From the point of view of the German Animal Welfare Federation the following issues are to be requested: (1)In order to enable a fast and comprehensive advancement of promising genetechnological non-animal test methods, it should be ensured that public funding is provided with an adequate budget and over a sufficiently long period of time. (2)The legislator should initiate broad discussions on the question if society would be willing to dispense with certain pieces of knowledge if they would necessarily have to be gained at the expense of a certain degree of animal suffering. As the case may be, in the German Animal Welfare Act it should be laid down that certain procedures should not be considered acceptable as such. (3)As long as animal experiments with transgenic animals continue to be performed, concrete legal measures should be laid down in the German Animal Welfare Act to ensure that the distress of the animals (taking into account all factors relevant for transgenic animals) and the expected benefit of the research project are determined objectively so that the outcome of the ethical evaluation process becomes comprehensible. (4)The legislator should provide the authorities responsible for the licensing of research projects with concrete instructions in order to ensure that all aspects relevant for the welfare of the animals are fully taken into account when evaluating the ethical acceptability and scientific indispensability of projects and that special attention is given to research projects with transgenic animals. (5)The German Decree on the Reporting of Laboratory Animals should be amended to ensure that all individual transgenic animals are included in the official statistical reports regardless of whether they end up being used in scientific procedures or not. From the point of view of animal welfare it is possible to redesign biomedical research to do without transgenic animals without impeding necessary scientific progress. The survey in hand sought to make a contribution to providing a scientifically sound background for initiating these discussions. PMID- 16477342 TI - Effort thrombosis of the upper extremities related to an arm stretching exercise. AB - "Effort" axillary-subclavian vein thrombosis (Paget-Schroetter syndrome) is an uncommon deep venous thrombosis due to repetitive activity of the upper limbs. Most cases of this condition are related to strenuous or prolonged sport or occupational activities, while others are associated with endogenous or exogenous risk factors. We report the case of a 43-year-old, previously healthy, male who developed right axillary-subclavian venous thrombosis, which was possibly associated with an exercise involving arm extension and shaking in a posture of shoulder abduction and outstretched for 10 minutes on 2 consecutive days. The condition improved but returned with increased severity when he resumed the exercise after a 2-day break, when he presented with a swollen and bluish arm at the emergency department. Sonographic examination showed moderate thrombotic stenosis of the right axillary vein. Effort thrombosis was diagnosed after ruling out associated coagulopathy or concomitant malignancy. External compression of the accessory ribs or lymph nodes were not detected. He was treated with low molecular weight heparin, followed by oral anticoagulant therapy for 6 months. Only partial resolution of thrombosis was achieved after 6 months of anticoagulant therapy, but pulmonary embolism did not occur during 18 months of follow-up. This case illustrates that, although unusual, Paget-Schroetter syndrome can occur in a healthy patient as a result of mild to moderate exercise. PMID- 16477344 TI - Introducing the concept of the 3Rs into tissue engineering research. AB - Tissue engineering, defined as using a combination of cultured cells and biodegradable scaffolds to repair tissue damaged by injury or disease, represents a booming sector of biomedical research. Animal experimentation is routinely performed prior to clinical trials. The presented study tries to translate the aspect of the 3Rs to tissue engineering research: Cell culture protocols were adapted to antibiotic free and serum free conditions. Biomaterials (Bio-Gide and a collagen sponge prototype) were pre-tested using the HET-CAM assay. CAM-testing suggested a protocol change for application of the Bio-Gide scaffold and demonstrated unsuitable material properties of the collagen sponge. Application of 3R compliant protocols for tissue engineering research led to increased cell proliferation, higher synthesis of extracellular matrix molecules, reduced dedifferentiation and more information about the biomaterials at an early experimental stage. Tissue engineering research can therefore profit from the increased efforts to validate in vitro alternatives and supplements to animal testing. PMID- 16477345 TI - [Replacement of dogs as research animals for the approval testing of plant protection products]. AB - The replacement of animal testing using dogs for the registration of plant protection products requires a long-term step-by-step procedure. The first goal should be to achieve international agreement on using only one single study in dogs. This would result in a significant short-term reduction of the use of dogs for this purpose. The competent working groups both in the EU and the United States EPA have declared this to be their intended aim. In this context, the 90 day study is to be the preferred study from the scientific as well as the animal welfare points of view. It is proposed to set up an international expert task force within the next 12 months, which should seek to initiate a process of international harmonization of the testing requirements following the example of the International Conference of Harmonization of Technical Requirements for Medical Products, ICH. The goal should be to achieve international agreement on only one single study with dogs within the next 2 to 3 years. In addition, other valid scientific procedures, with which the use of dogs for testing can be reduced, should be critically assessed. A complete replacement of the use of dogs for plant protection product testing is suggested to take place at a later stage. This may be achieved by either deriving safety threshold values by applying a safety factor to chronic NOAEL values obtained in studies using rats for those groups of substances, for which there is evidence that the dog is the more sensitive species, or by combining the chronic rat study with other animal tests stipulated for the registration of pesticides. PMID- 16477346 TI - [The role of animal testing advisory committees in biomedical research in Germany]. AB - In accordance with the German Animal Welfare Act, animal experiments in fundamental biomedical research may only be performed after licensing by the responsible authority. This license may only be granted if the experiments are considered indispensable and if the distress of the animals seems ethically acceptable in relation to the purpose of the study. Since 1987 advisory committees have been established to support the authorities in the evaluation of these provisions. Animal welfare organisations had expected case-by-case evaluations of the in-dispensability of research proposals and of the distress of the animals and the scientific benefit of the experiments to take place in these committees, so that such projects that would not meet the criteria of ethical acceptability could be prevented. However, already the lack of parity in the advisory committees alone, in which as a rule four scientists counterpart two representatives from animal welfare organisations, often-times prevents a balanced discussion of these provisions from taking place. Additionally, due to the freedom of science granted in the German Constitution without reservations, until 2002 also the licensing authorities were merely permitted to perform a formal examination of the applications. In the mean time, by including animal welfare as a national objective in the Constitution, the preconditions were made to enable an examination of the contents. From the point of view of animal welfare it therefore is to be requested that now also the advisory committees are ascribed more importance in the course of the licensing procedure and to establish the legal framework for this, if necessary by a revision of the Animal Welfare Act. PMID- 16477347 TI - [Ablation profiles in corneal laser surgery. Current and future concepts]. AB - The predictability and quality of results in corneal refractive laser surgery are determined by a number of factors. Here, the calculation and choice of the ablation profile represent central elements. Our growing knowledge about the physical and optical properties of the eye in recent years has led to the development of different strategies in the generation of ablation profiles. This review describes the currently used ablation profiles with their advantages and disadvantages and provides an outlook on future methods for the calculation of ablation profiles. PMID- 16477348 TI - Microbial reductive dechlorination of weathered and exogenous co-planar polychlorinated biphenyls (PCBs) in an anaerobic sediment of Venice Lagoon. AB - The occurrence of reductive dechlorination processes towards pre-existing PCBs and five exogenous coplanar PCBs were investigated in a contaminated sediment of Porto Marghera (Venice Lagoon, Italy) suspended, under strictly anaerobic conditions, in water collected from the same site. PCB dechlorination started after five months of incubation, when sulfate initially occurring in the microcosms was completely depleted and methanogenesis was in progress. It was ascribed to sulfate-reducing bacteria. Several pre-existing hexa-, penta- and tetra-chlorinated biphenyls were slowly bioconverted into tri- and di-, ortho substituted PCBs from the 5th to the 16th month of experiment. Spiked coplanar PCBs, i.e., 3,3',4,4'-tetrachlorobiphenyl, 3,3',4,4',5- and 2,3',4,4',5 pentachlorobiphenyls, 3,3',4,4',5,5'- and 2,3,3',4,4',5-hexachlorobiphenyls, were extensively transformed (by about 90%) into lower chlorinated congeners, such as 3,3',5,5'-/2,3',4,4'-tetrachlorobiphenyl, 3,3',5-, 2,4,4'-, 2,3',4- and 2,3',5 trichlorobiphenyl, 3,4-/3,4'- and 3,3'-dichlorobiphenyl and 2-chlorobiphenyl. The reductive dechlorination of spiked PCBs did not influence significantly the biotransformation rate and extent of pre-existing PCBs. PMID- 16477349 TI - The contribution of water soluble and water insoluble organic fractions to oxygen uptake rate during high rate composting. AB - This study aims to establish the contribution of the water soluble and water insoluble organic fractions to total oxygen uptake rate during high rate composting process of a mixture of organic fraction of municipal solid waste and lignocellulosic material. This mixture was composted using a 20 l self-heating pilot scale composter for 250 h. The composter was fully equipped to record both the biomass-temperature and oxygen uptake rate. Representative compost samples were taken at 0, 70, 100, 110, 160, and 250 h from starting time. Compost samples were fractionated in water soluble and water insoluble fractions. The water soluble fraction was then fractionated in hydrophilic, hydrophobic, and neutral hydrophobic fractions. Each fraction was then studied using quantitative (total organic carbon) and qualitative analysis (diffuse reflectance infrared spectroscopy and biodegradability test). Oxygen uptake rates were high during the initial stages of the process due to rapid degradation of the soluble degradable organic fraction (hydrophilic plus hydrophobic fractions). Once this fraction was depleted, polymer hydrolysis accounted for most of the oxygen uptake rate. Finally, oxygen uptake rate could be modeled using a two term kinetic. The first term provides the oxygen uptake rate resulting from the microbial growth kinetic type on easily available, no-limiting substrate (soluble fraction), while the second term considers the oxygen uptake rate caused by the degradation of substrate produced by polymer hydrolysis. PMID- 16477350 TI - Characterisation of biodegradation capacities of environmental microflorae for diesel oil by comprehensive two-dimensional gas chromatography. AB - In contaminated soils, efficiency of natural attenuation or engineered bioremediation largely depends on biodegradation capacities of the local microflorae. In the present study, the biodegradation capacities of various microflorae towards diesel oil were determined in laboratory conditions. Microflorae were collected from 9 contaminated and 10 uncontaminated soil samples and were compared to urban wastewater activated sludge. The recalcitrance of hydrocarbons in tests was characterised using both gas chromatography (GC) and comprehensive two-dimensional gas chromatography (GCxGC). The microflorae from contaminated soils were found to exhibit higher degradation capacities than those from uncontaminated soil and activated sludge. In cultures inoculated by contaminated-soil microflorae, 80% of diesel oil on an average was consumed over 4-week incubation compared to only 64% in uncontaminated soil and 60% in activated sludge cultures. As shown by GC, n-alkanes of diesel oil were totally utilised by each microflora but differentiated degradation extents were observed for cyclic and branched hydrocarbons. The enhanced degradation capacities of impacted-soil microflorae resulted probably from an adaptation to the hydrocarbon contaminants but a similar adaptation was noted in uncontaminated soils when conifer trees might have released natural hydrocarbons. GCxGC showed that a contaminated-soil microflora removed all aromatics and all branched alkanes containing less than C(15). The most recalcitrant compounds were the branched and cyclic alkanes with 15-23 atoms of carbon. PMID- 16477351 TI - Growth rate and nutrient limitation affect the transport of Rhodococcus sp. strain DN22 through sand. AB - Rhodococcus strain DN22 grows on the nitramine explosive RDX as a sole nitrogen source, and is potentially useful for bioremediation of explosives-contaminated soil. In order for strain DN22 to be effectively applied in situ, inoculum cells must reach zones of RDX contamination via passive transport, a process that is difficult to predict at field-scale. We examined the effect of growth conditions on the transport of DN22 cells through sand columns, using chemostat-grown cultures. Strain DN22 formed smaller coccoid cells at low dilution rate (0.02 h( 1)) and larger rods at high dilution rate (0.1 h(-1)). Under all nutrient limitation conditions studied, smaller cells grown at low dilution rate were retained more strongly by sand columns than larger cells grown at high dilution rate. At a dilution rate of 0.05, cells from nitrate-limited cultures were retained more strongly than cells from RDX-limited or succinate-limited cultures. Breakthrough concentrations (C/C (0)) from sand columns ranged from 0.04 (nitrate limited, D=0.02 h(-1)) to 0.98 (succinate-limited, D=0.1 h(-1)). The observed strong effect of culture conditions on transport of DN22 cells emphasizes the importance of physiology studies in guiding the development of bioremediation technologies. PMID- 16477352 TI - Immobilized cell reactors in mineralization of dicarboxylic acid solid waste. AB - Dicarboxylic acid solid waste containing phthalic acid, malic acid, quinone, saturated and unsaturated dicarboxylic esters etc., are discharged in huge quantities during the crackdown of benzene over the catalyst vanadium at temperatures greater than 500 degrees C in a dicarboxylic acid manufacturing industry. Concern over the biological effects of these compounds underlines the necessity to treat this solid waste. The role of yeast Saccharomyces cerevisiae and anaerobic mixed bacterial cultures immobilized in activated carbon, in sequential two stage anoxic reactors, were investigated for the degradation of dicarboxylic acid solid waste (DASW). In the first stage, DASW was dissolved in water to yield a concentration of 0.5% w/v and was treated in yeast Saccharomyces cerevisiae immobilized reactor at an optimum residence time of 24 h. The yeast fermented samples were further treated in an upflow anaerobic reactor containing mixed culture immobilized in activated carbon at an Hydraulic Retention Time (HRT) of 0.2076 days at an hydraulic flow rate of 14.6x10(-3 )m(3)/day and Chemical Oxygen Demand (COD) loading rate of 4.3 kg/m(3)/day. The intermediates that were formed during the yeast fermentation and the anaerobic degradation of DASW were characterized by HPLC, proton NMR, C(13) NMR and mass spectrometry. PMID- 16477353 TI - A bioremediation approach using natural transformation in pure-culture and mixed population biofilms. AB - Bacterial transformation by naked DNA is thought to contribute to gene transfer and microbial evolution within natural environments. In nature many microbial communities exist as complex assemblages known as biofilms where genetic exchange is facilitated. It may be possible to take advantage of natural transformation processes to modify the phenotypes of biofilm communities giving them specific and desirable functions. Work described here shows that biofilms composed of either pure cultures or mixed populations can be transformed with specific catabolic genes such that the communities acquire the ability to degrade a particular xenobiotic compound. Biofilms were transformed by plasmids bearing genes encoding green fluorescent protein (mut2) and/or atrazine chlorohydrolase (atzA). Confocal microscopy was used to quantify the number of transformants expressing mut2 in the biofilms. Degradation of atrazine by expressed atzA was quantified by tandem mass spectrometry. PCR analysis was performed to confirm the presence of atzA in transformed biofilms. These results indicate that it should be possible to use natural transformation to enhance bioremediation processes performed by biofilms. PMID- 16477354 TI - Molecular characterization of microbial populations at two sites with differing reductive dechlorination abilities. AB - This study compares three molecular techniques, including terminal restriction fragment length polymorphism (T-RFLP), RFLP analysis with clone sequencing, and quantitative PCR (Q-PCR) for surveying differences in microbial communities at two contaminated field sites that exhibit dissimilar chlorinated solvent degradation activities. At the Idaho National Engineering and Environmental Laboratory (INEEL), trichloroethene (TCE) was completely converted to ethene during biostimulation with lactate. At Seal Beach, California, perchloroethene (PCE) was degraded only to cis-dichloroethene (cDCE) during biostimulation but was degraded to ethene after bioaugmentation with a dechlorinating culture containing Dehalococcoides strains. T-RFLP analysis showed that microbial community composition differed significantly between the two sites, but was similar within each site among wells that had low or no electron donor exposure. Analysis of INEEL clone libraries by RFLP with clone sequencing revealed a complex microbial population but did not identify any Dehalococcoides strains. Q PCR targeting the 16S rRNA gene of Dehalococcoides strains - known for their unique capability to dechlorinate solvents completely to ethene - revealed a significant population at INEEL, but no detectable population at Seal Beach prior to bioaugmentation. Detection of Dehalococcoides by Q-PCR correlated with observed dechlorination activity and ethene production at both sites. Q-PCR showed that Dehalococcoides was present in even the pristine well at INEEL, suggesting that the difference in dechlorination ability at the two sites was due to the initial absence of this genus at Seal Beach. Of the techniques tested, Q PCR quantification of specific dechlorinating species provided the most effective and direct prediction of community dechlorinating potential. PMID- 16477355 TI - Organic compounds in re-circulated leachates of aerobic biological treated municipal solid waste. AB - Biodegradation of organic matter is required to reduce the potential of municipal solid waste for producing gaseous emissions and leaching contaminants. Therefore, we studied leachates of an aerobic-treated waste from municipal solids and a sewage sludge mixture that were re-circulated to decrease the concentration of biodegradable organic matter in laboratory-scale reactors. After 12 months, the total organic C and biological and chemical oxygen demands were reduced, indicating the biodegradation of organic compounds in the leachates. Curie-point pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS) and pyrolysis-field ionization mass spectrometry (Py-FIMS) revealed that phenols, alkylaromatic compounds, N-containing compounds and carbohydrates were the predominate compounds in the leachates and solid waste. Leachate re-circulation led to a higher thermal stability of the residual organic matter as indicated by temperature-resolved Py-FIMS. Admixture of sewage sludge to solid waste was less effective in removing organic compounds from the leachates. It resulted in drastic higher and more bio-resistant loads of organic matter in the leachates and revealed increased proportions of alkylaromatic compounds. The biodegradation of organic matter in leachates, re-circulated through municipal solid waste, offers the potential for improved aerobic waste treatments and should be investigated on a larger scale. PMID- 16477356 TI - Cloning of the organophosphorus pesticide hydrolase gene clusters of seven degradative bacteria isolated from a methyl parathion contaminated site and evidence of their horizontal gene transfer. AB - Seven organophosphorus pesticide-degrading bacteria harboring the methyl parathion degrading (mpd) gene were isolated from a methyl parathion contaminated site. In this study, the 4.7 kb mpd gene cluster, conserved in all seven bacteria capable of degrading methyl parathion, was cloned and further analysis revealed that this cluster contained five ORFs and the mpd gene was associated with a mobile element, IS6100. In addition to mpd gene ORF and tnpA ORF, three other ORFs showed high homology to the permease component of ABC-type transport system, the general secretion pathway protein B, and the RNA polymerase sigma 70 factor, respectively. The mpd genes of these 7 strains were subcloned and expressed in E. coli, SDS-PAGE and zymogram analysis showed that two expression products of mpd genes in E. coli were found, but the one without signal peptide showed the hydrolytic activities. Our evidences collectively suggest that mpd gene cluster may be disseminated through horizontal gene transfer based on phylogenetic analysis of the cluster and their host bacterial strains, and comparisons of GC content of the cluster and respective host's chromosome. PMID- 16477357 TI - The influence of structural components of alkyl esters on their anaerobic biodegradation in marine sediment. AB - Ester-based organic compounds are one type of synthetic base fluid added to drilling mud used during off-shore oil-drilling operations in the Gulf of Mexico. Concern over the environmental impact of synthetic base fluid (SBF) contaminated rock cuttings discharged into the Gulf of Mexico has prompted the promulgation of EPA regulations requiring that all SBF be tested for biodegradability in marine sediment prior to their use in the Gulf. In order to allow the design or selection of suitably biodegradable esters, the anaerobic biodegradability of a variety of ester compounds was tested using a marine sediment inoculum to reveal the effect of: (a) increasing the chain length of the acid moiety, (b) increasing the chain length of the alcohol moiety; (c) alternating the relative size of the alcohol and acid moieties, (d) branching in the alcohol moiety, and (e) the presence of an unsaturated bond in the acidic moiety. The chemical structure of esters was found to affect the completeness and rate of anaerobic biodegradation, and would affect their ability to be certified for use as an SBF in the Gulf of Mexico. Recommendations for ester usage include using esters that have a total carbon number of between 12 and 18 and avoiding the use of branched alcohols (or acids by inference). The presence of an unsaturated bond in the acid (or alcohol by inference) increased biodegradability of the ester. PMID- 16477358 TI - Effect of carbon starvation on toluene degradation activity by toluene monooxygenase-expressing bacteria. AB - Subsurface bacteria commonly exist in a starvation state with only periodic exposure to utilizable sources of carbon and energy. In this study, the effect of carbon starvation on aerobic toluene degradation was quantitatively evaluated with a selection of bacteria representing all the known toluene oxygenase enzyme pathways. For all the investigated strains, the rate of toluene biodegradation decreased exponentially with starvation time. First-order deactivation rate constants for TMO-expressing bacteria were approximately an order of magnitude greater than those for other oxygenase-expressing bacteria. When growth conditions (the type of growth substrate and the type and concentration of toluene oxygenase inducer) were varied in the cultures prior to the deactivation experiments, the rate of deactivation was not significantly affected, suggesting that the rate of deactivation is independent of previous substrate/inducer conditions. Because TMO-expressing bacteria are known to efficiently detoxify TCE in subsurface environments, these findings have significant implications for in situ TCE bioremediation, specifically for environments experiencing variable growth-substrate exposure conditions. PMID- 16477359 TI - Influence of a nonaqueous phase liquid (NAPL) on biodegradation of phenanthrene. AB - A series of batch reactor experiments was carried out to examine the effect of a nonaqueous phase liquid (NAPL) on the biodegradation of a hydrophobic solute. A mathematical program model that describes physical processes of solute solubilization and partitioning between the NAPL and aqueous phases as well as microbial degradation and oxygen utilization was used to analyze the test data. The model calculates the cumulative changes in concentration of substrate, cell mass, carbon dioxide, and dissolved oxygen as a function of time. The equations incorporate the effects of solute solubilization, partitioning, biodegradation, as well as oxygen availability. Hexadecane was used as the model NAPL and was not biodegraded in the timeframe of the experiments performed. The model solute was the polyaromatic hydrocarbon, phenanthrene. In agreement with several previous studies, experimental measurements showed that hexadecane increased rates of mineralization of 15 mg phenanthrene when present at low mass but decreased rates at high mass. Model results suggest that partitioning of the phenanthrene into the hexadecane phase limits bioavailability at high NAPL mass. Further the model suggests that mineralization rates were higher with the low NAPL mass because aqueous phenanthrene concentrations were higher in those treatments from ca. 20 to 40 h than in other treatments. Finally, experiments showed that the presence of hexadecane, at all masses tested, resulted in a lower cell yield, effectively increasing the amount of CO(2) produced during the experiment. Model results suggest that this is due to changes in phenanthrene metabolism that are induced by the presence of the hexadecane phase. Model studies aimed at increasing rates of biodegradation by modifying operating conditions are described along with practical approaches to implementing these modifications. PMID- 16477360 TI - An investigation into the microbial clogging potential of selected filter media as a result of biodegradation of a high-strength sulphate-rich alkaline leachate. AB - The research examines the potential for bio-clogging in filter packs containing fine sand of the type typically used in extraction wells for pumping leachates containing fine particulate matter, such as cement kiln dust (CKD). Three filter media with different particle sizes were used: 1.7-4.75, 0.35-1.0, and 0.235-0.45 mm. Each sand filter was tested using a leachate recirculating column reactor with a free drainage layer, on top of which was placed the filtration medium which was kept saturated and at a positive hydrostatic head by a 2-l reservoir of leachate. The leachate was collected from a landfill site that had been used for the co-disposal of municipal solid waste (MSW) and CKD. The leachate used was filtered by passing through a Whatman GFA filter paper before being added to the reactors in order to eliminate as far as possible the non-biological clogging which might have resulted from the introduction of particulate matter in the form of CKD. The filters and a control experiment were run under anaerobic conditions at 35 degrees C. The bio-clogging potential was observed by taking differential manometer readings from manometers located in the drainage and reservoir sections of the reactor. No clogging was detected using the coarser of the filter media, but there was some clogging when a finer filter medium was used. Head space gas analysis indicated that methanogenic activity was inhibited and analysis of the liquid phase indicated that the microbial process responsible for removal of chemical oxygen demand (COD) was principally one of sulphate reduction. PMID- 16477361 TI - Microbial decolorization of reactive black-5 in a two-stage anaerobic-aerobic reactor using acclimatized activated textile sludge. AB - A two-stage anaerobic-aerobic treatment process based on mixed culture of bacteria isolated from textile dye effluent was used to degrade reactive black 5 dye (RB-5). The anaerobic step was studied in more detail by varying the dye concentration from 100 to 3000 mg l(-1). The results showed that major decolorization was achieved during the anaerobic process. The time required for decolorization by > 90% increased as the concentration of the dye increased. It was also found that maintaining dissolved oxygen (DO) concentration below 0.5 mg l(-1 )and addition of a co-substrate viz., glucose, facilitates anaerobic decolorization reaction remarkably. An attempt was made to identify the metabolites formed in anaerobic process by using high performance liquid chromatography (HPLC) and UV-VIS spectrophotometry. A plate assay was performed for the detection of dominant decolorizing bacteria. Only a few bacterial colonies with high clearing zones (decolorization zones) were found. The results showed that under anaerobic condition RB-5 molecules were reduced and aromatic amines were generated. The aromatic amine metabolite was partly removed in subsequent aerobic bio-treatment. It was possible to achieve more than 90% decolorization and approximately 46% reduction in amine metabolite concentration through two-stage anaerobic-aerobic treatment after a reaction period of 2 days. PMID- 16477362 TI - Differential responses of a mine tailings Pseudomonas isolate to cadmium and lead exposures. AB - We examined cadmium and lead resistance in Pseudomonas sp. S8A, an isolate obtained from mine tailings-contaminated soil. Resistant to soluble metal concentrations up to 200 mg l(-1) cadmium and 300 mg l(-1) lead, S8A produced both exopolymer and biosurfactant. Upon growth, this pseudomonad diverged into two morphologically distinct colony subtypes; small and round or large and flat. In the presence of lead and in the no metal control the large morphotype appeared only in late stationary phase. With cadmium the large morphotype appeared immediately following exposure. Results show that the large morphotype produced greater amounts of surfactant than the small morphotype, suggesting a unique subpopulation response to cadmium toxicity. Results also indicate that an unidentified 28 kDa protein was expressed following exposure to >10 mg l(-1) cadmium. This study demonstrates new links between surfactant production, differential subpopulation response and metal exposure. PMID- 16477363 TI - Modelling MSW decomposition under landfill conditions considering hydrolytic and methanogenic inhibition. AB - A landfill typically progresses through a series of microbial degradation phases, in which hydrolysis, production and consumption of fermentation products, such as fatty acids, and methane formation play important roles. For ultimate degradation of the waste, stable methanogenic conditions have to be attained, and maintained for sufficient time. Using experimental data from 100-L landfill simulation reactors containing municipal solid waste from a residential area, a distributed model, which accounts for vertical water flow, was developed. As a first step, the waste was divided into two fractions: readily degradable and recalcitrant waste. Secondly, the general hydrolysis of the recalcitrant waste was accounted for by including a specific, well-defined chemical substance in the model that generally occurs in Municipal Solid Waste (MSW) and is hydrolysed before its further degradation to methane. For this purpose we chose diethyl phthalate and its hydrolysis product monoethyl phthalate, for which leachate data are available from the reactors. The model indicated that inhibition of the hydrolytic and methanogenic processes occurred during the acidogenic phase and that it could be overcome either by improving the chemical environment or by the complete oxidation of the inhibiting, i.e. the easily degraded, fraction of the waste. The generality of the model was confirmed by the patterns of the phthalate di- and monoester transformations obtained. The validity of the model was further confirmed using experimental data from parallel reactors, which were subjected to either leachate exchange with an already methanogenic reactor or to initial aeration to force the reactor into stable methanogenic conditions. PMID- 16477365 TI - A novel compound heterozygous mutation in the thyroglobulin gene resulting in congenital goitrous hypothyroidism with high serum triiodothyronine levels. AB - Thyroglobulin abnormality is a rare cause of congenital hypothyroidism and only a limited number of mutations in the thyroglobulin gene have been reported. We analyzed the thyroglobulin gene in a patient with congenital goitrous hypothyroidism. This girl was identified with hyperthyrotropinemia in a neonatal mass-screening test. The patient had goiter, and her body weight gain was poor. Distal femoral epiphysis was absent on roentgenography. Her serum thyroxine level was low; however, her triiodothyronine level was high. Autoantibodies against triiodothyronine, thyroid peroxidase, and thyroglobulin were all negative. Her serum thyroglobulin level was undetectable. The thyroglobulin gene from the genomic DNA of the patient was analyzed by direct sequencing. Two novel heterozygous missense mutations, Cys1897Tyr (exon 31) and Arg2336Gln (exon 40), were found in the patient. The former mutation was derived from her mother, suggesting a compound heterozygous state. Normal triiodothyronine and low thyroxine concentrations are often observed in patients with thyroglobulin gene mutations. We considered that some patients with thyroglobulin abnormality might have high triiodothyronine levels. In cases of congenital goitrous hypothyroidism with normal-to-high triiodothyronine levels and low serum thyroglobulin levels, thyroglobulin abnormality should be considered. PMID- 16477364 TI - The unique characteristics of Thai Leber hereditary optic neuropathy: analysis of 30 G11778A pedigrees. AB - Leber hereditary optic neuropathy (LHON) is characterized by acute or subacute bilateral visual loss, and affects mostly young males. The most common mitochondrial DNA mutation responsible for LHON worldwide is G11778A. Despite different genetic backgrounds, which are believed to influence the disease expression, most features of LHON are quite common in different populations. However, there seem to be a few ethnic-specific differences. Analyses of our 30 G11778A LHON pedigrees in Thailand showed some characteristics different from those of Caucasians and Japanese. In particular, our pedigrees showed a lower male to female ratio of affected persons (2.6:1) and much higher prevalence of G11778A blood heteroplasmy (37% of the pedigrees contained at least one heteroplasmic G11778A individual). Heteroplasmicity seemed to influence disease manifestation in our patients but did not appear to alter the onset of the disease. The estimated overall penetrance of our G11778A LHON population was 37% for males and 13% for females. When each of our large pedigrees were considered separately, disease penetration varied from 9 to 45% between the pedigrees, and also varied between different branches of the same large pedigree. Survival analysis showed that the secondary LHON mutations G3316A and C3497T had a synergistic deleterious effect with the G11778A mutation, accelerating the onset of the disease in our patients. PMID- 16477368 TI - The effect of vitamin A pretreatment on radiation induced alteration in neutrophil functions. AB - The aim of this investigation was to determine whether pre-administration of vitamin A will be effective in preventing the radiation-induced decline in MPO H2O2 system and the end product of reactive nitrogen species (NOx) in guinea pig. Animals were subjected to 612 cG of radiation and polimorfonuclear leukocytes were isolated and then NOx and myeloperoxidase activity were measured. In irradiated animals, a marked decrease in NOx level and myeloperoxidase activity have been found compared to control (p = 0.001 and p < 0.000 respectively). The application of vitamin A significantly improved the radiation-induced decrease (for both p < 0.00). In conclusion pre-treatment of vitamin A is efficient to protect against radiation induced alteration in polimorfonuclear leukocyte. PMID- 16477366 TI - A principal role for AtXTH18 in Arabidopsis thaliana root growth: a functional analysis using RNAi plants. AB - Rearrangement of cellulose microfibrils within cell-wall matrices is considered one of the most critical steps in the regulation of both the orientation and extent of cell expansion in plants. Xyloglucan endotransglucosylase/hydrolases (XTHs) are a family of enzymes that mediate the construction and restructuring of load-bearing cross links among cellulose microfibrils. The Arabidopsis thaliana XTH genes AtXTH17, 18, 19, and 20 are phylogenetically closely related to one another and are preferentially expressed in the roots. However, they exhibit different expression profiles within the root and respond to hormonal signals differently. To investigate their functions in root growth, we examined phenotypes of loss-of-function mutants for these genes using T-DNA insertion lines and RNAi plants. These functional analyses disclosed a principal role for the AtXTH18 gene in primary root elongation. Of the four XTH genes, AtXTH18 exhibits the highest level of mRNA expression. We also determined auxin-signaling pathways for these genes using a mutant with a defect in the AXR2/IAA7 gene and found that the expression of AtXTH19 in the elongation/maturation region of the root is under the control of the AXR2/IAA7 signaling pathway. PMID- 16477367 TI - Ingestion of difructose anhydride III partially restores calcium absorption impaired by vitamin D and estrogen deficiency in rats. AB - BACKGROUND: Vitamin D (VD) and estrogen deficiencies impair Ca absorption and bone mineralization, and the relevance of the interaction between these factors has not been fully understood. AIM OF THE STUDY: The aim of the present study was to clarify the effects of a nondigestible saccharide, difructose anhydride III (DFAIII), on the interaction of VD and estrogen deficiencies involved in Ca malabsorption by assessing changes in intestinal Ca absorption and bone mineralization by feeding of DFAIII in rats with VD or estrogen deficiency or with a combined loss of VD and estrogen. METHODS: Three-week-old female Sprague Dawley rats were divided into four groups: two groups were ovariectomized (OVX) and two were laparotomized (sham). One group each of OVX and sham rats were fed an AIN93G-based normal diet, and the other groups were fed a VD-deficient diet for 8-weeks. Rats from the four groups were divided into two subgroups and fed the normal or VD-deficient diet with or without DFAIII for next 4-weeks. RESULTS: VD deficiency decreased Ca absorption and bone mineralization with reductions in duodenal calbindin D9k mRNA and serum Ca levels. There were no additional reductions in these parameters in the OVX. The reductions in Ca absorption and femoral Ca were restored partially or fully by DFAIII. Recovery of Ca absorption rate by DFAIII was greater in the OVX than in the sham showing an interaction between OVX and VD deficiency in, at least, the DFAIII-fed groups. The cecal pH was lower and the level of short-chain fatty acids in the cecal contents was higher in all the DFAIII groups than those in the control groups. CONCLUSIONS: VD deficiency impaired Ca absorption and bone mineralization, and feeding DFAIII partially restored Ca malabsorption and fully recovered bone Ca in VD-deficient rats. No additional reductions in these parameters with a combination of VD deficiency and OVX were noted. However, interactions were found between these factors in the DFAIII-induced increase in Ca absorption. PMID- 16477369 TI - Preferential radioprotection to DNA of normal tissues by ferulic acid under ex vivo and in vivo conditions in tumor bearing mice. AB - Our previous study showed that ferulic acid (FA) offered good radioprotection under in vitro and in vivo conditions to DNA and enhanced the DNA repair process in the peripheral blood leucocytes of mice in vivo. This study concerns radioprotection of normal versus tumor cells. Administration of FA (50 mg/kg body weight) to mice bearing fibrosarcoma tumor, 1 h prior to/ or immediately after radiation exposure (4 Gy) showed preferential radioprotection to normal cells i.e. peripheral blood leucocytes and bone marrow cells in comparison to tumor cells. This preferential protection under in vivo conditions could be attributed to poor vasculature in the tumor or peculiar characteristics of the tumor cells either to restrict its entry inside the cells or metabolize or inactivate the drug. To resolve these ex vivo study was carried out using bone marrow and tumor cells. It was found that under ex vivo condition also only bone marrow cells were protected by FA. Thus the studies revealed that FA showed preferential protection to normal cells under both in vivo and ex vivo conditions. PMID- 16477370 TI - Expression of the ErbB4 receptor causes reversal regulation of PP2A in the Shc signal transduction pathway in human cancer cells. AB - Expression of ErbB4 receptor is correlated with the incidence of non-metastatic types of human cancers, whereas the overexpression of other ErbB receptor families (ErbB1/EGFR, ErbB2 and ErbB3) is correlated to the formation of metastatic tumors. However, the molecular mechanism underlying this phenomenon has been unclear. Earlier, we demonstrated that okadaic acid (OA), an inhibitor of a serine/threonine phosphatase PP2A, stimulated the growth hormone-induced ERK phosphorylation in the wild type Chinese hamster ovary (CHO) cells and the cells expressing ErbB1 receptor, but suppressed ERK activation in CHO cells that express ErbB4 receptor. PP2A had been understood as a negative regulator of the growth hormone-stimulated signal transduction pathways, however, this observation suggested that expression of ErbB4 receptor reversed the regulation of PP2A in the ErbB4 signalling pathway. In this study, we found that OA suppressed phosphorylation of Shc at Tyr317, therefore it down-regulated ERK phosphorylation in the ErbB4 expressing CHO cells. Accordingly, basal PP2A contributed to the phosphorylation of Shc Tyr317 in ErbB4 expressing CHO cells, nevertheless it had been reported that PP2A negatively regulates Shc tyrosine phosphorylation in the EGF- or IGF-I-induced signalling pathways. By testing OA for human cancer cell lines that express different types of ErbB receptors, we found that ErbB4 receptor expression was accompanied with positive regulation of PP2A for phosphorylation of Shc Tyr317 and its downstream ERK phosphorylation in MCF-7 and SK-OV-3 cell lines, but not in LNCaP and PC-3 cells. Thus, PP2A regulates the ERK activity in a cell-specific manner, and it is speculated that distinct regulation of PP2A in the ErbB4 receptor signalling pathway may cause a difference in progression of cancer phenotypes. PMID- 16477372 TI - Brief antecedent anoxia preserves mitochondrial function after sustained undersupply: a subcellular correlate to ischemic preconditioning? AB - BACKGROUND: There is increasing evidence that mitochondria - owning a high degree of autonomy within the cell - might represent the target organelles of the myocardial protection afforded by ischemic preconditioning. It was the aim of the study to investigate a possible subcellular correlate to ischemic preconditioning at the mitochondrial level. In addition, we tested whether this protection depends on mitochondrial ATP-dependent potassium channels (K (ATP)) and an might involve an attenuation of mitochondrial ATP hydrolysis during sustained anoxia. METHODS AND RESULTS: Sustained anoxia (A, 14 min) and reoxygenation (R) completely inhibited state 3 and state 4 respiration of isolated ventricular mitochondria from Wistar rats. An antecedent brief anoxic incubation (4 min) followed by reoxygenation (2 min) prevented this loss of mitochondrial function. The protection afforded by anoxic preconditioning could be mimicked by the K (ATP) opener diazoxide (30 micromol/l) and was completely inhibited by the K (ATP) blocker 5-hydroxydecanoic acid (300 micromol/l). Structural mitochondrial integrity, as estimated from externalization of the mitochondrial enzymes creatine kinase and glutamateoxalacetate transaminase, remained unchanged between the groups, as did mitochondrial ATP loss during anoxia. CONCLUSION: For the first time, we provide direct evidence for a subcellular preconditioning-like functional mitochondrial adaptation to sustained anoxia. This effect apparently depends on opening of K(ATP) but is independent of ATP preservation. PMID- 16477371 TI - Docosahexaenoic acid and eicosapentaenoic acid-enriched phosphatidylcholine liposomes enhance the permeability, transportation and uptake of phospholipids in Caco-2 cells. AB - The influence of docosahexaenoic acid (DHA)- and eicosapentaenoic acid (EPA) enriched phosphatidylcholine (PC) on the permeability, transport and uptake of phospholipids was evaluated in Caco-2 cells. The cells were grown on permeable polycarbonate transwell filters, thus allowing separate access to the apical and basolateral chambers. The monolayers of the cells were used to measure lucifer yellow permeability and transepithelial electrical resistance (TEER). Transcellular transportation of diphenylhexatriene (DPH) labeled-PC small unilamellar vesicles (SUV) from the apical to basolateral chamber, and uptake of the same SUV was monitored in the cell monolayers. Cell-membrane perturbation was evaluated to measure the release of lactate dehydrogenase and to determine the cell viability with sodium 2-(4-iodophenyl)-3-(4-nitrophenyl) -5-(2, 4 disulfophenyl)-2H-tetrazolium dye reduction assay. The lucifer yellow flux was 1.0 and 1.5 nmol/h/cm(2) with 50 microM PC, and 17.0 and 23.0 nmol/h/cm(2) with 100 microM PC when monolayers of Caco-2 cells were treated with DHA- and EPA enriched PC, respectively. TEER decreased to 24 and 27% with 50 and 100 microM DHA-enriched PC, and to 25 and 30% with 50 and 100 microM EPA-enriched PC, respectively. Our results show that DHA- and EPA-enriched PC increases tight junction permeability across the Caco-2 cell monolayer whereas soy PC has no effect on tight junction permeability. Transportation and uptake of DHA- and EPA enriched PC SUV differed significantly (P < 0.01) from those of soy PC SUV at all doses. We found that PC SUV transported across Caco-2 monolayer and was taken up by Caco-2 cells with very slight injury of the cell membrane up to 100 microM PC. Lactate dehydrogenase release and cell viability did not differ significantly between the treatment and control, emphasizing that injury was minimal. Our results suggest that DHA- and EPA-enriched PC enhance the permeability, transport and uptake of PC SUV across monolayers of Caco-2 cells. PMID- 16477373 TI - Human placental protein/peptides stimulate melanin synthesis by enhancing tyrosinase gene expression. AB - Placental protein/peptides as biological response modifier are well documented, but not much known about melanogenesis. We possibly for the first time, demonstrated melanogenesis in B16F10 mouse melanoma by a placental protein/peptide fraction (PPPF) prepared from a hydroalcoholic extract of fresh term human placenta. This study described the effect of PPPF on the induction of tyrosinase; the key enzyme of melanogenesis to investigate the basis of PPPF induced pigmentation in primary melanocyte and B16F10 melanoma. Tyrosinase induction by PPPF in B16F10 cells was found dose- and time dependent at the level of activity. Tyrosinase, at the level of transcription and protein expression when assessed by RT-PCR and Western blot analyses found to have considerable induction over untreated control. PPPF led to enhanced activation of tyrosinase promoter resulting higher transcription thus substantiating the role of PPPF as a stimulator of melanogenesis. Actinomycin D, the transcriptional inhibitor of protein synthesis, blocked the stimulatory action of PPPF since the induction of tyrosinase and melanin was markedly reduced in presence of this inhibitor. Thus the results suggested that PPPF mediated increase in tyrosinase expression occurred through transcriptional upregulation to stimulate melanogenesis in B16F10 cells and in primary melanocyte also. PMID- 16477374 TI - Participation of estrogen receptors in the enhancement of osteoblast differentiation by TAK-778. AB - TAK-778 has been shown to stimulate osteogenesis both in vitro and in vivo. However, the mechanism by which TAK-778 exerts its effects is still unclear. There is evidence that TAK-778 acts via estrogen-receptor (ER)-mediated signaling; this study therefore aimed to investigate the roles that ERalpha, ERbeta, and membrane ER play in the osteogenic effect of TAK-778. To this end, human bone marrow mesenchymal cells were cultured with TAK-778 in the presence of either ICI182,780 (ERalpha and ERbeta antagonist) or MPP (ERalpha antagonist) or PD98059 (an extracellular-regulated kinase inhibitor that acts on the membrane ER pathway). The following parameters were evaluated: cell proliferation, collagen content, alkaline phosphatase (ALP) activity and bone-like formation. Data were compared using ANOVA. The effect of TAK-778 on expression of ERalpha and ERbeta was investigated by immunolabeling. In order to investigate whether TAK-778 binds to ER, an ER binding assay was performed. Both immunolabeling and binding assays were conducted using cells from human alveolar bone. The osteogenic effect of TAK 778 was inhibited by ICI182,780 and MPP; however, it was not affected by PD98059. The expression of both ERalpha and ERbeta was not affected by TAK-778. The competition curve obtained from the binding assay using TAK-778 showed maximal displacement when 10(-5) M TAK-778 was used. This study's results show that TAK 778 enhances osteoblast differentiation through an ERalpha-dependent pathway by binding to this receptor and not by increasing the expression of ER. PMID- 16477376 TI - Characterization of thymocyte phenotypic alterations induced by long-lasting beta adrenoceptor blockade in vivo and its effects on thymocyte proliferation and apoptosis. AB - Adult male Wistar rats were subjected to propranolol (P, 0.40 mg/100 g/day) or saline (S) administration (controls) over 14 days. The expression of major differentiation molecules on thymocytes and Thy-1 (CD90) molecules, which are shown to adjust thymocyte sensitivity to TCRalphabeta signaling, was studied. In addition, the sensitivity of thymocytes to induction of apoptosis and concanavalin A (Con A) signaling was estimated. The thymocytes from P-treated (PT) rats exhibited an increased sensitivity to induction of apoptosis, as well as to Con A stimulation. Furthermore, P treatment produced changes in the distribution of thymocyte subsets suggesting that more cells passed positive selection and further differentiated into mature CD4+ or CD8+ single positive (SP) TCRalphabeta(high) cells. These changes may, at least partly, be related to the markedly increased density of Thy-1 surface expression on TCRalphabeta(low) thymocytes from these rats. The increased frequency of cells expressing the CD4+25+ phenotype, which has been shown to be characteristic for regulatory cells in the thymus, may also indicate alterations in thymocyte selection following P treatment. Inasmuch as positive and negative selections play an important role in continuously reshaping the T-cell repertoire and maintaining tolerance, the hereby presented study suggests that pharmacological manipulations with beta-AR signaling, or chemically evoked alterations in catecholamine release, may interfere with the regulation of thymocyte selection, and consequently with the immune response. PMID- 16477375 TI - Is there a specific role for the plasma membrane Ca2+ -ATPase in the hepatocyte? AB - The plasma membrane Ca2+ -ATPase (PMCA) is responsible for the fine, long-term regulation of the cytoplasmic calcium concentration by extrusion of this cation from the cell. Although the general kinetic mechanisms for the action of both, well coordinated hydrolytic activity and calcium transport are reasonably understood in the majority of cell types, due to the complex physiologic and biochemical characteristics shown by the hepatocyte, the study of this enzyme in this cell type has become a real challenge. Here, we review the various molecular aspects known to date to be associated with liver PMCA activity, and outline the strategies to follow for establishing the role of this enzyme in the overall physiology of the hepatocyte. In this way, we first concentrate on the basic biochemical aspects of liver cell PMCA, and place an important emphasis on expression of its molecular forms to finally focus on the critical hormonal regulation of the enzyme. Although these complex aspects have been studied mainly under normal conditions, the significance of PMCA in the calcium homeostasis of an abnormal liver cell is also reviewed. PMID- 16477377 TI - Multilineage differentiation of adipose-derived stromal cells from GFP transgenic mice. AB - Functional engineering of musculoskeletal tissues generally involves rapid expansion of progenitor cells in vitro while retaining their potential for further differentiation and then induction in specific culture conditions. The autologous adipose-derived stromal cells (ASCs) are considered to contain pluripotent mesenchymal stem cells. Imaging with expression of green fluorescent protein (GFP) facilitates the detailed research on ASCs physiological behavior during differentiation into a variety of cell lineages both in vitro and in vivo. In this study, we aimed to confirm the trans-germ plasticity of homogeneously marked ASCs from GFP transgenic mice. Simultaneously, the term and intensity of GFP expression in ASCs were also focused on during variant inductions, when cells were incubated with multiple growth factors and adjuvant. ASCs were harvested from inguinal fat pads of transgenic nude mice, passaged 3 times in monolayer cultures, and then transferred to osteogenic, adipogenic, neurogenic, and myogenic medium. The morphological characterization of inductive cells was observed using phase-contrast microscopy and histological staining such as alizarin red for mineralization nodules and oil red O for lipid accumulation. The expression of marker genes or proteins was measured using RT-PCR and immunocytochemical analysis. Collagen type I, osteopontin (OPN), and osteocalcin (OCN) were positive in osteogenic lineages, peroxisome proliferator-activated receptor(PPAR)-gamma2 and lipoprotein lipase (LPL) were positive in adipogenic ones, glial fibrillary acidic protein (GFAP) and neuron-specific enolase (NSE) were positive in neurogenic ones, and alpha-smooth muscle actin (alpha-SMA) was positive in myogenic ones. Moreover, the results of fluorescence microscopic imaging suggested that there was no significant decline of GFP expression during ASCs differentiation and the level of GFP maintained stable till differentiated ASCs showed apoptotic phenotype. So the endogenous GFP and multilineage potential of transgenic ASCs had no influences on each other. Since the population of GFP ASCs can be easily identified, it is proposed that they may be promising candidate seed cells for further studies on ASCs tissue engineering, especially the study on engineered tissues formed in vivo. PMID- 16477378 TI - Age-related differences in myocardial hydrogen ion buffering during ischemia. AB - BACKGROUND: During myocardial ischemia, accumulation of end products from anaerobic glycolysis (hydrogen ions (H(+)), lactate) can cause cellular injury, consequently affecting organ function. The cells' ability to buffer H(+) (buffering capacity (BC)) plays an important role in ischemic tolerance. Age related differences in myocardial lactate and H(+) accumulation (one hour of ischemia) as well as differences in BC, myoglobin (Mb) and histidine (His) contents in the left (LV) and right (RV) ventricles were assessed in neonatal compared to adult pigs. The BC of the septum was also compared. METHODS AND RESULTS: Neonatal RV and LV had lactate accumulations of 43% and 63% and significantly greater H(+) (p < 0.004) compared to the adult. In the neonate LV, BC was 17% significantly poorer (p = 0.0001), had 33% lower Mb (p = 0.0002) and 15% lower His content (p = 0.0004) when compared to the adult. In the RV, despite similar BC between the neonate and adult, myoglobin content was 36% (p = 0.0004) lower in the neonate. The neonate septum had a BC that was 11% lower than that of the adult. With maturation, the adult LV had a BC that was 10% greater (p < 0.01) than the RV while the septum mirrored that of the LV. CONCLUSIONS: During maturation to adulthood, the BC of the septum begins to closely resemble the LV. Neonatal hearts have a potentially greater vulnerability to acid-base disturbances during ischemia in both ventricles when compared to hearts of adults. This is due to lower levels of myoglobin and histidine in the young, which could render them more susceptible to injury during ischemia. During myocardial ischemia, H(+) and lactate accumulation may pose deleterious effects on the heart. The ability to buffer H(+) (buffering capacity, BC) affects ischemic tolerance. Although lactate accumulation during 1 h of global ischemia was similar between ventricles of neonatal and adult swine, H(+) accumulation was greater and BC, Mb and His content were lower. With maturation, LV BC was higher than the RV while septum developmentally resembled the LV. Thus, hearts of neonates may be at a greater risk of ischemic injury compared to hearts of adults. PMID- 16477379 TI - Sodium butyrate induces apoptosis and cell cycle arrest in primary effusion lymphoma cells independently of oxidative stress and p21(CIP1/WAF1) induction. AB - Primary effusion lymphoma, a peculiar type of B cell non-Hodgkin lymphoma, preferentially develops in immunodeficient individuals and its pathogenesis is closely linked with human herpesvirus 8 (HHV-8). HHV-8 is present primarily persistence in primary effusion lymphoma cells, and the lytic cycle of HHV-8 can be induced by sodium butyrate (NaB) treatment. HHV-8 gene expression is affected by NaB in BCBL-1 cells, but the cellular response of BCBL-1 cells upon NaB treatment has not been investigated to date. Using BCBL-1 cells, a HHV-8 harboring cell line, we demonstrated that sodium butyrate could induce the reactive oxygen species generation, apoptosis and cell cycle arrest in BCBL-1 cells. The sodium butyrate-induce cell cycle arrest was associated with the decrease of Cdc2, Cdk4 and cyclin A in BCBL-1 cells without altering the protein levels of p21(CIP1/WAF1). The apoptosis induced by sodium butyrate in BCBL-1 cells was independent of oxidative stress. PMID- 16477381 TI - Translocation of long chain fatty acids across the plasma membrane--lipid rafts and fatty acid transport proteins. AB - Translocation of long chain fatty acids across the plasma membrane is achieved by a concert of co-existing mechanisms. These lipids can passively diffuse, but transport can also be accelerated by certain membrane proteins as well as lipid rafts. Lipid rafts are dynamic assemblies of proteins and lipids, that float freely within the two dimensional matrix of the membrane bilayer. They are receiving increasing attention as devices that regulate membrane function in vivo and play an important role in membrane trafficking and signal transduction. In this review we will discuss how lipid rafts might be involved in the uptake process and how the candidate proteins for fatty acid uptake FAT/CD36 and the FATP proteins interact with these domains. We will also discuss the functional role of FATPs in general. To our understanding FATPs are indirectly involved in the translocation process across the plasma membrane by providing long chain fatty acid synthetase activity. PMID- 16477380 TI - Peroxisome proliferator-activated receptor-alpha selective ligand reduces adiposity, improves insulin sensitivity and inhibits atherosclerosis in LDL receptor-deficient mice. AB - Fenofibrate, a selective (1)PPAR-alpha activator, is prescribed to treat human dyslipidemia. The aim of this study was to delineate the mechanism of fenofibrate mediated reductions in adiposity, improvements in insulin sensitivity, and lowering of triglycerides (TG) and free fatty acids (FFA) and to investigate if these favorable changes are related to the inhibition of lipid deposition in the aorta. To test this hypothesis we used male LDLr deficient mice that exhibit the clinical features of metabolic syndrome X when fed a high fat high cholesterol (HF) diet. LDLr deficient mice fed HF diet and simultaneously treated with fenofibrate (100 mg/kg body weight) prevented development of obesity, lowered serum triglycerides and cholesterol, improved insulin sensitivity, and prevented accumulation of lipids in the aorta. Lowering of circulating lipids occurred via down-regulation of lipogenic genes, including fatty acid synthase, acetyl CoA carboxylase and diacyl glycerol acyl transferase-2, concomitant with decreased liver TG and cholesterol, and TG output rate. Fenofibrate also suppressed liver apoCIII mRNA levels and markedly increased lipoprotein lipase mRNA levels, known to enhance serum TG catabolism. In addition, fenofibrate profoundly reduced epididymal fat and mesenteric fat mass to the levels seen in lean mice. The reductions in body weight were associated with elevation of hepatic uncoupling protein 2 (UCP2) mRNA, a concomitant increase in the ketone body formation, and improved insulin sensitivity associated with tumor necrosis factor-alpha reductions and phosphoenol pyruvate carboxykinase down-regulation. These results demonstrate that fenofibrate improves lipid abnormalities partly via inhibition of TG production and partly via clearance of TG-rich apoB particles by elevating LPL and reduced apoCIII. The prevention of obesity development occurred via energy expenditure. Fenofibrate-mediated hypolipidemic effects together with improved insulin sensitivity and loss of adiposity led to the reductions in the aortic lipid deposition by inhibiting early stages of atherosclerosis possibly via vascular cell adhesion molecule-1 (VCAM-1) modulation. These results suggest that potent PPAR-alpha activators may be useful in the treatment of syndrome X. PMID- 16477382 TI - Whole body exposure to low frequency magnetic field: no provable effects on the cellular energetics of rat skeletal muscle. AB - On the basis of previous experience with biological effects of electromagnetic fields a potential effect of homogeneous sinusoidal magnetic field (50 Hz, 10 mT) on energy state of rat skeletal muscle was investigated. Two different total body exposures to magnetic field were selected: (1) repeated 1 hour exposure, 2 times a week for 3 months, and (2) acute 1.5 hour exposure (and the appropriate control groups). Important energy metabolites (adenosine triphosphate--ATP, creatine phosphate, creatine, lactate, pyruvate and inorganic phosphate) were analysed by enzymatic and spectroscopic methods in musculus gracilis cranialis. On the basis of the concentration of important energy metabolites the apparent Gibbs free energy of ATP hydrolysis and creatine charge was calculated. Our results demonstrate no influence of this low frequency magnetic field on the level of important energy metabolites in rat skeletal muscle. The conclusion of this study is that neither repeated exposure nor the acute exposure of rats to the sinusoidal magnetic field of given parameters has any important influence on the energy state of the skeletal muscle. PMID- 16477383 TI - Oxalate mediated nephronal impairment and its inhibition by c-phycocyanin: a study on urolithic rats. AB - The assumption of oxidative stress as a mechanism in oxalate induced renal damage suggests that antioxidants might play a beneficial role against oxalate toxicity. An in vivo model was used to investigate the effect of C-phycocyanin (from aquatic micro algae; Spirulina spp.), a known antioxidant, against calcium oxalate urolithiasis. Hyperoxaluria was induced in two of the 4 groups of Wistar albino rats (n = 6 in each) by intraperitoneally injecting sodium oxalate (70 mg/kg body weight). A pretreatment of phycocyanin (100 mg/kg body weight) as a single oral dosage was given, one hour prior to oxalate challenge. An untreated control and drug control (phycocyanin alone) were employed. Phycocyanin administration resulted in a significant improvement (p < 0.001) in the thiol content of renal tissue and RBC lysate via increasing glutathione and reducing malondialdehyde levels in the plasma of oxalate induced rats (p < 0.001), indicating phycocyanin's antioxidant effect on oxalate mediated oxidative stress. Administering phycocyanin after oxalate treatment significantly increased catalase and glucose-6-phosphate dehydrogenase activity (p < 0.001) in RBC lysate suggesting phycocyanin as a free radical quencher. Assessing calcium oxalate crystal retention in renal tissue using polarization microscopy and renal ultrastructure by electron microscopy reveals normal features in phycocyanin-- pretreated groups. Thus the study presents positive pharmacological implications of phycocyanin against oxalate mediated nephronal impairment and warrants further work to tap this potential aquatic resource for its medicinal application. PMID- 16477384 TI - Modulation of DNA intercalation by resveratrol and genistein. AB - Time correlated Single Photon Counting study (TCSPC) was performed for the first time to evaluate the effect of resveratrol (RES) and genistein (GEN) at 10-100 microM and 10-150 microM respectively, in modulating the DNA conformation and the variation induced due to intercalation by the dyes, ethidium bromide (EtBr) and acridine orange (AO). It is demonstrated using UV-absorption and fluorescence spectroscopy that RES and GEN, at 50 microM and 100 microM respectively can bind to DNA resulting in significant de-intercalation of the dyes, preventing their further intercalation within DNA. Hyperchromicity with red/blue shifts in DNA when bound to dyes was reduced upon addition of RES and GEN. DNA-dependent fluorescence of EtBr and AO was quenched in the presence of RES by 87.97% and 79.13% respectively, while similar quenching effect was observed for these when interacted with GEN (85.52% and 83.85%). It is found from TCSPC analysis that the higher lifetime component or constituent of intercalated dyes (tau(2), A (2)) decreased with the subsequent increase in smaller component or constituent of free dye (tau(1), A (1)) after the interaction of drugs with the intercalated DNA. Thus these findings signify that RES and GEN can play an important role in modulating DNA intercalation, leading to the reduction in DNA-directed toxicity. PMID- 16477385 TI - Effects of administration of Embelin and Curcumin on lipid peroxidation, hepatic glutathione antioxidant defense and hematopoietic system during N nitrosodiethylamine/Phenobarbital-induced hepatocarcinogenesis in Wistar rats. AB - The effects of administration of Embelin (EMB) and Curcumin (CUR) on lipid peroxidation, hepatic glutathione antioxidant defense and hematopoietic cells were examined during N-nitrosodiethylamine (DENA-200 mg kg(-1)body wt, single I.P injection) initiated and Phenobarbital (PB-0.05% in drinking water orally for 13 weeks) promoted hepatocarcinogenesis in Wistar strain male albino rats. DENA/PB induced hepatic damage was manifested by a significant drop in the hepatic glutathione antioxidant defense, increased lipid peroxidation and histological alterations like dysplasia, and atypical cells with abnormal chromatin pattern. Treatment with Curcumin (100 mg kg(-1)body wt) and Embelin (50 mg kg(-1)body wt) prevented the drop in hepatic glutathione antioxidant defense, decreased lipid peroxidation, minimized the histological alterations induced by DENA/PB, but showed toxic effects on the hematopoietic cells. Results indicate the beneficial effects of Embelin and Curcumin against oxidative tissue damage during chemically induced hepatocarinogenesis in rats. PMID- 16477386 TI - New oxo-bridged peroxotungsten complexes containing biogenic co-ligand as potent inhibitors of alkaline phosphatase activity. AB - Novel dinuclear peroxo complexes of tungsten with coordinated cystine of the type A(2)[W(2)O(3)(O(2))(4)(cystine)].4H(2)O, A = Na (1) or K (2) have been synthesized from the reaction of A(2)WO(4,)cysteine and 30% H(2)O(2)at pH 2.5. The synthesized compounds were characterized by elemental analysis, spectral and physico-chemical methods. The two W(VI) centres with side-on bound peroxo groups of the dinuclear complex species are bridged by an oxo group and a cystine ligand, formed from the oxidation of cysteine. Cystine occurring as zwitterion binds the metal centers of the complex ion through O(carboxylate) atoms leading to hepta co-ordination around each W(VI). The compounds exhibit high stability toward decomposition in solution of acidic as well as physiological pH and serve as weak substrates to catalase, undergoing degradation in presence of the enzyme at a rate much slower relative to H(2)O(2). The compounds efficiently oxidized GSH to GSSG, a reaction in which only two of the peroxide groups of the complex species were found to participate. The compounds induce strong inhibitory effect on alkaline phosphatase activity with a potency higher than that of the free cystine, tungstate, or peroxotungstate. PMID- 16477387 TI - Obese state leads to elevated levels of TGF-beta and COX isoforms in platelets of Zucker rats. AB - Platelets are rich sources of growth factors and enzymes that are implicated in a number of diseases including obesity, atherosclerosis, heart disease, syndrome X, liver and kidney diseases and certain types of cancers. In this research we investigated, if platelets in Zucker obese rats differ from their lean counterparts with respect to the levels of TGF-beta and COX isoforms, implicated in the pathogenesis of chronic diseases. In addition, we investigated if energy intake of the animals affects the platelet physiology. Platelets were isolated from obese and lean rats bearing preneoplastic lesions in their colon. Prior to platelet isolation these rats were fed either ad libitum (Ob or Ln) or energy restricted (Ob-ER or Ln-ER) diets for 8 weeks (n = 8/group). The levels of TGF beta1/-beta2 and COX-1/-2 proteins in platelets were analyzed by Western blot. The platelets of the Ob rats had significantly higher levels of TGF-beta1, COX-1/ 2 (p < 0.001) than did the platelets of the Ln rats and were not affected by moderate energy restriction. There were no significant differences in the protein expression of platelet TGF-beta2 among any of the groups. These results demonstrate that cytokines and candidates playing a role in the pathogenesis of chronic diseases, such as TGF-beta1 and COX-1/-2, are over-expressed in platelets of Zucker obese rats by comparison to their lean counterparts. These findings also demonstrate that the genotype of the animals exerts a significant effect on the biochemical composition of the platelets and could contribute to the pathogenesis of colon cancer and other metabolic abnormalities associated with obesity. PMID- 16477388 TI - Increase in extracellular cross-linking by tissue transglutaminase and reduction in expression of MMP-9 contribute differentially to focal segmental glomerulosclerosis in rats. AB - Tissue transglutaminase (tTG) is a Ca(2+)-dependent enzyme which stabilizes the extracellular matrix (ECM) through post-translational modification, and may play an important role in the pathogenesis of focal and segmental glomerulosclerosis (FSGS). Here, we have investigated whether tTG contributes to the glomerular ECM expansion in the puromycin aminonucleoside (PAN)-injection-induced experimental rat model of FSGS. The localization and expression of tTG, MMP-9 gelatinase, and the ECM component fibronectin (FN) in kidneys was determined by immunohistochemistry and measured by semi-quantitative analysis. Protein levels of tTG and MMP-9 were also analyzed by Western blotting.In situtransglutaminase activity was assayed by measurement of incorporated substrate and the immunofluorescence staining for the cross-linking product, epsilon-(gamma glutamyl) lysine. Prominent proteinuria, a typical pathological feature of FSGS, was observed in PAN injection group rats. tTG immunoreactivity was located markedly in glomeruli and the levels of this protein in whole-kidney homogenates of PAN injection group rats were significantly increased (361+/- 106% control, P< 0.05). Similarly, transglutaminase activity and epsilon-(gamma-glutamyl) lysine were also predominately located within glomeruli and were much more intense in the PAN-injected group than that in control animals. MMP-9 was also located primarily within glomeruli. In PAN-injected kidneys, protein levels of active MMP 9 were significantly reduced (59+/- 27% control, P< 0.01), while pro-MMP-9 levels increased (148+/- 42% control, P< 0.05). Remarkable expression of glomerular fibronectin (FN) was found in PAN injection group rats. Semi-quantitative analysis demonstrated this increased intensity of FN staining in the PAN-injected rats was 149+/- 23% of the control values (P< 0.05). Enhanced cross-linking of ECM by tissue transglutaminase and decreased degradation due to reduced active MMP-9 expression may be at least partially responsible for the deposition of FN within injured glomeruli in experimental FSGS. PMID- 16477389 TI - Effects of lactate dehydrogenase suppression and glycerol-3-phosphate dehydrogenase overexpression on cellular metabolism. AB - In order to conduct a physiological functional study of lactate dehydrogenase (LDH) and glycerol-3-phosphate dehydrogenase (GPDH), we engineered a CHO dhfr(-) cell, by overexpressing either the anti-sense LDH-A RNA (anti-LDH cells) or GPDH (GP3 cells), or both (GP3/anti-LDH cells). LDH activity in the cell cytosol, and lactate content and pHe change in the growth media were found to decrease according to the order: cell lines GP3/anti-LDH > anti-LDH > GP3 > CHO. Intracellular ATP contents, representing the extent of respiration rate, also decreased, according to a rank order as follows: GP3 > CHO > GP3/anti-LDH > anti LDH. We also attempted to identify and characterize any physiological changes occurring in the cells which harbored diverse metabolic pathways. First, anti-LDH cells with heightened respiration rates were found to display a higher degree of sensitivity to the prooxidant tert-butyl hydroperoxide (tBOOH), and the mitochondrial complex III inhibitor, antimycin A, than the GPDH-expressing cells (GP3 and GP3/anti-LDH), which have a lower respiration rate. Second, the anti sense LDH-A RNA-expressing cells (anti-LDH and GP3/anti-LDH) evidenced a higher degree of resistance to apoptosis by cell-cell contact inhibition, and a faster doubling time ( approximately 19 h compared with approximately 26 h) than the CHO and GP3 cells. Additionally, cell growth in an extended culture under HCO(3) (-) free conditions to induce a steep acidification could be maintained with the anti sense LDH-A RNA-expressing cells, but could not be maintained with the CHO and GP3 cells. Third, we observed that the most appropriate cell line for the optical production of a certain therapeutic protein (Tissue-Plasminogen Activator) was the GP3/anti-LDH cells. Collectively, our data indicate a variety of physiological roles for LDH and GPDH, including cellular acidosis, oxidoresistance, apoptosis by both acidosis and cell-cell contact inhibition, cell growth, and the generation of recombinant proteins. PMID- 16477391 TI - Circular dichroism and cross-linking studies of bacteriorhodopsin mutants. AB - Circular dichroism (CD) spectroscopy was employed for native (wild type, WT) bacteriorhodopsin (bR) and several mutant derivatives: R134K, R134H, R82Q, S35C, L66C, and R134C/E194C. Comparative analysis of the CD spectra in visible range shows that only R134C/E194C exhibits biphasic CD, typical for native bR, the other mutants demonstrate CD spectra with significantly smaller or absent negative band. Since the biphasic CD is a feature of hexagonal lattice structure composed by bR trimers in the purple membrane, these mutants and WT were examined by cross-linking studies, which confirmed the same trend towards trimeric organization. Therefore, a single amino acid substitution may lead to drastically different CD spectra without disruption of bR trimeric organization. Thus, although disruption of bR trimeric crystalline lattice structure (e.g., solubilization with detergents) directly results in the disappearance of characteristic bilobe in visible CD, the lack of the bilobe in the CD alone does not predict the absence of trimers. PMID- 16477390 TI - Formation and positioning of nucleosomes: effect of sequence-dependent long-range correlated structural disorder. AB - The understanding of the long-range correlations (LRC) observed in DNA sequences is still an open and very challenging problem. In this paper, we start reviewing recent results obtained when exploring the scaling properties of eucaryotic, eubacterial and archaeal genomic sequences using the space-scale decomposition provided by the wavelet transform (WT). These results suggest that the existence of LRC up to distances approximately 20-30 kbp is the signature of the nucleosomal structure and dynamics of the chromatin fiber. Actually the LRC are mainly observed in the DNA bending profiles obtained when using some structural coding of the DNA sequences that accounts for the fluctuations of the local double-helix curvature within the nucleosome complex. Because of the approximate planarity of nucleosomal DNA loops, we then study the influence of the LRC structural disorder on the thermodynamical properties of 2D elastic chains submitted locally to mechanical/topological constraint as loops. The equilibrium properties of the one-loop system are derived numerically and analytically in the quite realistic weak-disorder limit. The LRC are shown to favor the spontaneous formation of small loops, the larger the LRC, the smaller the size of the loop. We further investigate the dynamical behavior of such a loop using the mean first passage time (MFPT) formalism. We show that the typical short-time loop dynamics is superdiffusive in the presence of LRC. For displacements larger than the loop size, we use large-deviation theory to derive a LRC-dependent anomalous-diffusion rule that accounts for the lack of disorder self-averaging. Potential biological implications on DNA loops involved in nucleosome positioning and dynamics in eucaryotic chromatin are discussed. PMID- 16477394 TI - Uptake and elimination of lead, zinc, and copper by caddisfly larvae (Trichoptera: Hydropsychidae) using stable isotope tracers. AB - Stable isotopes of Pb, Zn, and Cu were used in laboratory experiments to determine the uptake and elimination of these metals by stream-dwelling caddisfly (Trichoptera: Hydropsychidae) larvae. For Pb and Cu, larvae were exposed to environmentally realistic levels (2.5 and 4.5 microg x L(-1), respectively) of one isotope for 9 days followed by a 9-day exposure to either the same isotope, to a second stable isotope of the same metal, or to RW containing no added isotope (two phases in total). For zinc, the exposure concentration was 15 microg x L(-1), and the experiment lasted for a total of 27 (i.e., three phases) rather than 18 days to see if uptake and elimination changed during the extended time period. The uptake clearances (k(u)) determined for the various metals averaged 7.8, 1.4, and 0.6 L x g dw(-1) x d(-1) for Pb, Zn, and Cu, respectively, if the total metal concentration in the water was used in the calculations. The clearance rate constants (k(e)) were less variable, averaging 0.15 d(-1) for Pb, 0.22 d(-1) for Zn, and approximately 0.1 d(-1) for Cu and were similar in both the presence (i.e., elimination) and absence (i.e., depuration) of metal in the water. These values are also comparable with those reported in the literature for other aquatic invertebrates. The use of stable isotopes thus allowed simultaneous measurement of uptake and clearance (elimination and depuration) of these metals at environmentally realistic concentrations and could be of great benefit for determining partitioning, assimilation efficiency, and pathways of these and other metals in the environment. PMID- 16477395 TI - Kinetic analysis on the skin disposition of cytotoxicity as an index of skin irritation produced by cetylpyridinium chloride: comparison of in vitro data using a three-dimensional cultured human skin model with in vivo results in hairless mice. AB - PURPOSE: The aim of this study was to kinetically and dynamically analyze in vitro cytotoxicity as an index of skin irritation by use of a three-dimensional cultured human skin model and to compare the in vitro assay data with data from living animals. METHODS: A cationic surfactant, cetylpyridinium chloride (CPC), was selected as a model irritant. Living skin equivalent-high (LSE-high) and hairless mice were used for the in vitro and in vivo tests, respectively. Skin irritation dermatodynamics was evaluated by calorimetric thiazoyl blue (MTT) conversion assay both for in vitro and in vivo tests, whereas dermatokinetics of CPC in LSE-high and mouse skin were evaluated using HPLC. RESULTS: The time course of cell viability in the skin after application of CPC to intact skin was distinctly different from that of stratum-corneum-stripped skin in both LSE-high and hairless mice. Biphasic behavior characterized by two first-order rates with an inflection time point was observed in intact skin, whereas cell viability monoexponentially decreased immediately after CPC application in stripped skin. The time courses of cell viability in the skin and dermatodynamics were closely related to that of dermatokinetics of CPC. CONCLUSION: The present study demonstrates that the in vitro cytotoxic profile was similar to the in vivo cytotoxicity test and that dermatodynamics was related to dermatokinetics of CPC. PMID- 16477396 TI - Report on the PQRI impurity characterization and quantification best practices survey. PMID- 16477398 TI - Prognosis and mortality of polymyositis and dermatomyositis patients. AB - The objective of this study was to assess the long-term outcome of polymyositis (PM) and dermatomyositis (DM) and the factors predictive of this outcome in a nationwide series in Finland. One hundred and seventy-six patients with PM and 72 patients with DM diagnosed in Finland in 1969-1985 were selected from the national hospital discharge register according to the diagnostic criteria of Bohan and Peter and followed up until death or till the end of August 1995. Gender, age, delay of therapy, serum creatine kinase, erythrocyte sedimentation rate, initial dose of corticosteroids and duration of cytostatic therapy were assessed as factors prognostic of death. The 5-year survival rate for PM was 75% [95% confidence interval (CI): 68-81%] and that for DM 63% (50-73%), and the respective 10-year survival rates were 55% (47-62%) and 53% (41-64%). The median survival for polymyositis was 11.0 years (95% CI: 9.5-13.3) and that for DM 12.3 years (5.5-20.7). The standardized mortality ratio for the combined group of PM and DM showed approximately threefold mortality compared to the general population. Compared to PM patients, the subjects with DM had a 1.47-fold (95% CI: 0.99-2.12) age- and sex-adjusted risk of mortality. Except for age in both groups and the delay in diagnosis in the PM group, no other individual factor reached significance as a predictor of death. However, cancer had a hazard ratio (HR) of 2.16 for death (95% CI: 0.95-4.50) in the DM group and 1.99 (95% CI: 1.01 3.94) in the PM group. A comparison of the causes of death in the PM and DM groups showed that the patients with DM had a greater risk of dying from cancer (age-adjusted HR 5.11, 95% CI: 2.31-11.3). According to this nationwide analysis of survival and its prognostic factors in patients with PM and DM, the latter group had an increased age-adjusted risk for mortality compared to the former. The difference seems to be mostly explained by the fact that the patients with DM had a higher risk of dying from cancer. PMID- 16477400 TI - Evaluation of left ventricular diastolic dysfunction with conventional and current Doppler techniques in Behcet's disease. AB - Behcet's disease (BD) is a systemic vasculitis characterized by genital and oral ulcers, uveitis, and other organs' involvement. Left ventricular (LV) diastolic dysfunction has been documented in BD. However, conventional echocardiographic techniques have serious limitations like its dependence on preload, afterload and heart rate. Recently, new techniques like colour M-mode and tissue Doppler imagining (TDI) have provided additional concept in the assessment of diastolic function. The aim of the present study was to investigate the LV diastolic dysfunction with conventional and new echocardiographic techniques in BD. Forty eight patients with BD (25 women, 23 men) and 26 healthy volunteers (15 women, 11 men) were enrolled in the study. LV diastolic functions were examined with mitral inflow pulse wave Doppler, TDI and mitral flow propagation rate (MFPR). The following were accepted as diastolic dysfunction: in mitral inflow pulse wave Doppler, E/A<1, isovolumic relaxation time (IVRT)>110 ms and deceleration time of E wave (DT)>240 ms; in TDI of mitral ring with pulse wave, E'/A'<1; and in MFPR, velocity slope (Vp)<45 cm/s. The two groups were comparable in age, sex, heart rate, body mass index, smoking, hyperlipidemia and basic echocardiographic measurements. LV diastolic dysfunction was significantly higher in BD group according to E/A<1 (p<0.05). When echocardiographic measurements were compared one by one for two groups, As' (late diastolic TDI wave in septal wall) was found to be significantly higher in BD group (p<0,0001). IVRT was longer in BD group than in controls, but it did not reach statistical significance (p=0,06). Diastolic dysfunction of LV is more frequent in patients with BD than in control according to E/A and As'. Conventional and current techniques like TDI and colour M-mode Doppler echocardiography could be used to investigate diastolic functions in BD. PMID- 16477399 TI - Evaluation of the Turkish version of the Bath Ankylosing Spondylitis Patient Global Score (BAS-G). AB - The objective of this study was to investigate the reliability and validity of the Turkish version of the Bath Ankylosing Spondylitis (AS) Patient Global Score (BAS-G). Seventy-one consecutive patients with AS were enrolled into the study. Patients were requested to fill in the questionnaire on the day of admission (first visit), on a second occasion within 24 h after admission (second visit) for test-retest reliability analysis, and on a third occasion for assessing sensitivity to change. Construct validity was assessed by correlation analysis with the Bath AS Functional Index (BASFI), Dougados Functional Index (DFI), Dougados Articular Index (DAI), physical examination findings, and several other parameters. Test-retest reliability analysis of individual BAS-G scores at initial and second visits showed good intraclass correlations [n=46, intraclass correlation=0.928 (0.870-0.960) and intraclass correlation=0.853 (0.725-0.920), for 1-week and 6-month scores, respectively]. Both 1-week and 6-month scores showed moderate correlations with the BASFI (r=0.586 and r=0.503, respectively, P=0.000 for both). The 1-week score also showed moderate correlation with the DFI (r=0.530, P=0.000). The 1-week score showed weak correlations with finger-to floor distance (r=0.263, P=0.027), chest expansion (r=-0.245, P=0.039), and DAI (r=0.271, P=0.036). Change in the 1-week score at the third visit showed good correlation with the BASFI score (r=0.670, P=0.000, n=36) and moderate correlation with the DFI (r=0.440, P=0.017, n=29). The Turkish version of the BAS G has good reliability and validity. It is a good tool for assessing patients with AS or other rheumatic diseases in clinical practice and research. PMID- 16477401 TI - Osteoarthritis is for the birds. AB - The purpose of this study was to assess birds as a potential model for osteoarthritis. Compromised by confounding factors, it has not been possible to clearly delineate causation in humans. Nonhuman mammals manifest osteoarthritis in the natural state too rarely for comparative study. Artificial environments (of captive animals) are associated with higher frequencies, but are still inadequate for comparative study, and surgical and chemical models provide only limited insight. As frequency of pathology (except trauma-related) in birds has not been systematically examined across species lines, several families were selected for examination. Skeletal collections of major museums were examined for presence of osteophytes in hawks and pigeons. Three percent of 2,243 free-ranging hawks and 9.8% of 2,718 pigeons had osteoarthritis, all localized to the ankle. The bird ankle morphologically resembles the human knee. Frequency in pigeons was significantly greater than in hawks (chi (2)=86.48, p<0.00001), but was no difference in frequency between wild caught and captive birds (chi (2)=1.06). While misconceptions have plagued past perspectives, it turns out that the most common form of arthritis in humans (osteoarthritis) is actually for, or at least, is common in the class Aves (birds). The frequency of osteoarthritis and cacophony of bird morphologies and behaviors provides an opportunity to start to understand such causation. PMID- 16477402 TI - Fat in the liver: diagnosis and characterization. AB - The purpose of this article is to provide an update on imaging techniques useful for detection and characterization of fat in the liver. Imaging findings of liver steatosis, both diffuse steatosis and focal fatty change, as well as focal fatty sparing, are presented. In addition, we will review computed tomography (CT) and magnetic resonance (MR) findings of focal liver lesions with fatty metamorphosis, including hepatocellular carcinoma, hepatocellular adenoma, focal nodular hyperplasia, angiomyolipoma, lipoma, and metastases. PMID- 16477403 TI - [Tumors of the craniocervical junction]. AB - In this review, the authors describe the most common tumors of the craniocervical junction and their management. Early recognition is important in view of the unspecific symptomatology and much better surgical results in patients operated before the appearance of neurological deficits. In the last 15 years, different craniobasal approaches have been developed which permit the radical excision of many of these lesions with no or acceptable morbidity. Representative cases are illustrated. PMID- 16477405 TI - Genetic diversity in seed populations of Echinacea purpurea controls the capacity for regeneration, route of morphogenesis and phytochemical composition. AB - The production of new varieties and higher quality products from Echinacea spp. requires a greater understanding of the regulation of plant growth and the production of specific phytometabolites. The current studies were designed to generate elite varieties of Echinacea purpurea based on regeneration efficiency and chemical profile. Clonal propagation of seedling-derived regenerants and screening for antioxidant potential and concentrations of caftaric acid, chlorogenic acid, cichoric acid, cynarin, and echinacoside identified 58 unique germplasm lines. Chemical profiles varied significantly among germplasm lines but were consistent within clones of each line. In temporary immersion bioreactors, exogenous application of the auxin indolebutyric acid significantly increased the cichoric acid and caftaric acid concentration in the root tissues. Together, these demonstrate the potential for selective breeding of elite, highly regenerative, chemically superior, clonally propagated varieties from the naturally occurring genetic variability in the seed populations of E. purpurea. PMID- 16477404 TI - Molecular cloning, expression and characterization of a glycosyltransferase from rice. AB - Secondary plant metabolites undergo several modification reactions, including glycosylation. Glycosylation, which is mediated by UDP-glycosyltransferase (UGT), plays a role in the storage of secondary metabolites and in defending plants against stress. In this study, we cloned one of the glycosyltransferases from rice, RUGT-5 resulting in 40-42% sequence homology with UGTs from other plants. RUGT-5 was functionally expressed as a glutathione S-transferase fusion protein in Escherichia coli and was then purified. Eight different flavonoids were used as tentative substrates. HPLC profiling of reaction products displayed at least two peaks. Glycosylation positions were located at the hydroxyl groups at C-3, C 7 or C-4' flavonoid positions. The most efficient substrate was kaempferol, followed by apigenin, genistein and luteolin, in that order. According to in vitro results and the composition of rice flavonoids the in vivo substrate of RUGT-5 was predicted to be kaempferol or apigenin. To our knowledge, this is the first time that the function of a rice UGT has been characterized. PMID- 16477406 TI - Isolation of cytoplasts from Satsuma mandarin (Citrus unshiu Marc.) and production of alloplasmic hybrid calluses via cytoplast-protoplast fusion. AB - Cytoplasm of Satsuma mandarin (Citrus unshiu Marc.) is known to influence seedlessness. Transfer of cytoplasm to a seedy cultivar could possibly lead to the production of seedless citrus fruits. In the present paper cytoplasts were isolated from cell suspension-derived protoplasts of Satsuma mandarin via ultra centrifugation in a discontinuous gradient. No nucleus could be detected in the cytoplasts by DAPI (4', 6-diamidino-2-phenylindole) staining compared with normal protoplasts. The cytoplasts, with high viability and small size, did not divide during solid embedding culture. Cytoplasts of Satsuma mandarin were electrically fused with embryogenic protoplasts of Murcott tangor (C. reticulata x C. sinensis), which led to regeneration of several cell lines. Flow cytometry (FCM) indicated that the cell lines were diploids. Simple sequence repeats (SSR) and cleaved amplified polymorphism sequence (CAPS) showed that the cell lines got their nuclear DNA from the protoplast parent, whereas the cytoplast parent donated the mtDNA, confirming transfer of mtDNA from Satsuma mandarin into Murcott tangor via cytoplast-protoplast fusion though no polymorphism was detected in chloroplast DNA between the fusion partners. This is the first report on isolation and characterization of cytoplasts, together with cytoplast protoplast fusion in Citrus, which has a potential for citrus cultivar improvement involving cytoplasm transfer via cytoplast-protoplast fusion. PMID- 16477407 TI - Rapid plant regeneration and analysis of genetic fidelity of in vitro derived plants of Chlorophytum arundinaceum Baker--an endangered medicinal herb. AB - An efficient in vitro multiplication system via multiple shoot bud induction and regeneration has been developed in Chlorophytum arundinaceum using shoot crown explants. Optimum regeneration frequency (87%) and desirable organogenetic response in the form of de novo organized multiple shoot buds without an intervening callus phase was obtained on Murashige and Skoog's (MS) minimal organics medium containing 3% sucrose (w/v) supplemented with 4 x 10(-6) M Kn and 2 x 10(-6) MIBA. Axenic secondary explants with multiple shoot buds on subculturing elicited best response with 1 x 10(-5) M Kinetin (Kn) and 5 x 10(-6) M indole-3-butyric acid (IBA) giving rise to an average of 18.74 shoots per culture with mean shoot length of 7.6 cm +/- 1.73. Varying molar ratios of either Kn/IBA or Kn/NAA revealed statistically significant differences in the regeneration frequencies among the phytohormone treatments. It was observed that the shoot bud differentiation and regeneration was influenced by the molar ratios of cytokinins/auxin rather than their relative concentrations. Healthy regenerated shoots were rooted in half strength MS basal medium containing 3% sucrose (w/v) supplemented with 5 x 10(-6) M IBA. Following simple hardening procedures, rooted plantlets, were transferred to soil-sand (1:1; v/v) with more than 90% success. Genetic fidelity was assessed using random amplified polymorphic DNA (RAPD), karyotype analysis and meiotic behaviour of in vitro and in vivo plants. Five arbitrary decamers displayed same banding profile within all the micropropagated plants and in vivo explant donor. The cytological and molecular analysis complemented and compared well and showed no genomic alterations in the plants regenerated through shoot bud differentiation. High multiplication frequency, molecular, cytological and phenotypic stability ensures the efficacy of the protocol developed for the production and conservation of this important endangered medicinal herb. PMID- 16477409 TI - Nocturnal melatonin concentration is correlated with illness severity in patients with septic disease. PMID- 16477408 TI - Dental anxiety in a representative sample of residents of a large German city. AB - In a demographic survey, 300 residents of a German city were questioned to determine the prevalence of dental anxiety. The correlation between the amount of dental anxiety and the age, sex, and education of the subjects was examined and the reasons for avoiding dentist's appointments, the duration of this avoidance, and what the subjects desire from future dental treatment. The Hierarchical Anxiety Questionnaire (HAQ) was used to measure the amount of dental anxiety. The average level of anxiety was 28.8+/-10.1 according to the HAQ. Young people were more afraid than older people (p=0.007), and women were more anxious than men (p=0.004). Of the women, 72% go to the dentist regularly, but only 60% of the men do (p=0.020). A painful experience while receiving dental treatment was given by 67% as the main reason for their dental anxiety, followed by a fear of needles (35%). The people wished for the most accurate information available about the dental treatment they receive (69%), followed by a compassionate dentist (62%), and treatment that is free of pain (62%). Of the people, 11% [95% CI: (7.5%; 14.5%)] suffer from dental phobia. All dental phobics were able to state the cause of their fear and more urgently wished for help from the dentist in overcoming their anxiety than the non-phobics (p=0.030). To satisfy the needs of the phobic patients, it appears necessary to screen the phobics out of the group of all patients and then offer them adequate therapy, or refer these patients to specialised treatment centres. PMID- 16477410 TI - Candida sp. isolated from bronchoalveolar lavage: clinical significance in critically ill trauma patients. AB - OBJECTIVE: Based on limited data, Candida sp. isolates from bronchoalveolar lavage (BAL) cultures in immunocompetent patients are thought to be contaminants rather than pathogens. The objective of this study was to determine the clinical significance of Candida sp. isolated from BAL cultures in critically ill trauma patients. DESIGN AND SETTING: Retrospective study in a level 1 trauma intensive care unit. PATIENTS AND PARTICIPANTS: All patients with Candida sp. isolated from BAL cultures over a 3-year period; 85 Candida positive BAL cultures from 62 patients were studied. MEASUREMENTS AND RESULTS: The primary outcomes were the incidence of Candida sp. in BAL, antifungal use, course of the possible infection, and mortality. Of 1077 BAL cultures 85 (8%) grew Candida sp., representing 64 episodes of possible Candida sp. ventilator-associated pneumonia. No colony counts exceeded the diagnostic threshold for bacterial VAP (>or=10(5) cfu/ml). Only 2 of 64 episodes (3%) were treated with systemic antifungals. Three other episodes (5%) were treated because of concomitant therapy for Candida sp. at other sites. The majority of episodes were not treated with antifungals and were considered contaminants (59/64, 92%). No patients developed subsequent candidemia, and most follow-up BALs (74%) were negative for Candida sp. Overall mortality (17%) was similar to previous patients with similar severity of injury at the study center (18%). CONCLUSIONS: The results of this study suggest that isolation of Candida sp. from BAL in quantities below the diagnostic threshold for VAP in this population does not require antifungal therapy. PMID- 16477411 TI - Saline volume in transvesical intra-abdominal pressure measurement: enough is enough. AB - OBJECTIVE: The objective was to determine the minimum volume of instillation fluid for intra-abdominal pressure (IAP) measurement, and to evaluate the effect of instillation volume on transvesically measured IAP. DESIGN: Prospective cohort study SETTING: Twenty-two-bed surgical ICU of the Ghent University Hospital. PATIENTS AND PARTICIPANTS: Twenty patients at risk of intra-abdominal hypertension (IAH). INTERVENTIONS: Transvesical IAP measurement using volumes from 10 to 100 ml. Minimal volume at which an IAP was measured was recorded (IAP(min)), as well as IAP at 50 and 100 ml of instillation volume (IAP(50) and IAP(100)). The percentage difference for IAP(50) and IAP(100) was calculated. MEASUREMENTS AND RESULTS: The minimal volume for IAP measurement was 10 ml in all patients. Mean IAP(min) was 12.8 mmHg (+/- 4.9), mean IAP(50 )15 mmHg (+/- 4.5) and mean IAP(100) 17.1mmHg (+/- 4.7). The mean percentage difference for IAP(50) was 21% (+/- 17%), and 40% (+/-29%) for IAP(100.) Twelve patients were categorised as suffering from IAH when 10 ml of saline was used for IAP measurement, increasing to 15 and 17 patients respectively when using 50 and 100 ml. In patients with IAH, there was a significant correlation between the duration of bladder drainage and percentage difference for IAP(100) (Pearson correlation coefficient 0.60, p = 0.03). CONCLUSIONS: Using 50 or 100 ml of saline for IAP measurement in critically ill patients results in higher IAP values compared with the use of 10 ml, and possibly, in overestimation of the incidence of intra-abdominal hypertension. PMID- 16477412 TI - Melatonin: possible implications for the postoperative and critically ill patient. AB - There is increasing interest in the hormone melatonin in postoperative and critically ill patients. The roles of melatonin in the regulation of the sleep wake cycle, resetting of circadian rhythm disturbances and its extensive antioxidant activity have potential applications in these patient groups. The interaction between melatonin and the stresses of surgery and critical illness are explored in the context of circadian rhythms, sleep disorders and delirium. The antioxidant activity is discussed in terms of the reduction of ischaemic reperfusion injury, prevention of multi-organ failure and treatment of sepsis. Unfortunately, there is currently insufficient evidence that exogenous melatonin is effective in preventing or treating postoperative delirium. Similarly, in the critically ill patient, sleep disorders are associated with disrupted melatonin circadian secretion, but there is a paucity of data to support routine exogenous melatonin supplementation. More clinical evidence to confirm the potential benefits of melatonin therapy is required before it can be routinely used in the postoperative or critically ill patient. PMID- 16477413 TI - Informed consent for research in ICU obtained before ICU admission. AB - OBJECTIVE: To analyze the procedure of the informed consent for ICU research obtained before ICU admission. DESIGN: Prospective, open, observational study. SETTING: 20-bed surgical ICU of a tertiary teaching university hospital and the ward before and after ICU. PATIENTS: Patients, scheduled for elective cardiac surgery, who accepted to participate in a coagulation study. INTERVENTIONS: Patients underwent the same informed consent procedure, including an oral presentation of the coagulation study and an informative leaflet the day before surgery on the ward. MEASUREMENTS AND RESULTS: Between January and August 2001, we included 38 patients; 36 survived ICU. Ten to 12 days after surgery, 8/36 (22%) patients did not know they had participated in a study, and 9/36 (25%) could not recall the study purpose and the related risk. Patients with incomplete recall stayed longer in ICU [median (range): 4 (3-6) vs 3 (1-5) days; p = 0.004]. None of these patients (0/9 vs 10/27; p < 0.04) had read the informative leaflet AND asked at least one question during the informed consent procedure. CONCLUSIONS: Even when the informed consent is obtained in the most optimal conditions for ICU research, its ethical value remains questionable. Indeed, a substantial number of patients were unaware of their study participation, or of the related purpose and risks. When the ICU stay is prolonged, we should at least repeatedly and actively (re)-inform patients about their study participation. PMID- 16477414 TI - Weight-adjusted resting energy expenditure is not constant in critically ill patients. AB - OBJECTIVE: In critically ill patients, energy requirements are frequently calculated as a multiple of total body weight presuming a linear relationship between total body weight and resting energy expenditure (REE); however, it is doubtful if this estimation of energy needs should be applied to all patients, particularly to overweight patients, since adipose tissue has a low contribution to REE. This study was undertaken to test the hypothesis that REE adjusted for total body weight decreases with increasing body mass index in critically ill patients. Additionally, measured REE was compared with three predictive equations. DESIGN AND SETTING: Clinical study in a university hospital intensive care unit. PATIENTS: One hundred critically ill patients admitted to the intensive care unit. MEASUREMENTS AND RESULTS: Patients were included into four groups according to their body mass index (normal weight, pre-obese, obese, and morbidly obese). Measured REE was assessed using indirect calorimetry. Energy needs were calculated using the basal metabolic rate, the Consensus Statement of the American College of Chest Physicians (REEacs), and 25[Symbol: see text]kcal/kg of ideal body weight (REEibw). Adjusted REE was 24.8 +/- 5.5 kcal/kg in normal weight, 22.0 +/- 3.7 kcal/kg in pre-obese, 20.4 +/- 2.6 kcal/kg in obese, and 16.3 +/- 2.3 kcal/kg in morbidly obese patients (p < 0.01). Basal metabolic rate underestimated measured REE in normal weight and pre-obese patients. REEacs and REEibw over- and underestimated measured REE in overweight patients, respectively. CONCLUSIONS: Predictive equations were not able to estimate measured REE adequately in all the patients. Adjusted REE decreased with increasing body mass index; thus, a body mass index group-specific adaptation for the estimation of energy needs should be applied. PMID- 16477415 TI - Mean glucose level is not an independent risk factor for mortality in mixed ICU patients. AB - OBJECTIVE: To find out if there is an association between hyperglycaemia and mortality in mixed ICU patients. DESIGN AND SETTING: Retrospective cohort study over a 2-year period at the medical ICU of a university hospital. MEASUREMENTS: Admission glucose, maximum and mean glucose, length of stay, mortality, insulin therapy and Apache-II score. RESULTS: In 1085 consecutive patients, ICU- and hospital mortality were 20 and 25%, respectively. The total number of blood glucose measurements was 10.012. Admission glucose was 7.9 +/- 4.5 mmol/l (mean +/- SD), mean glucose 7.5 +/- 2.9 and maximum glucose 10.0 +/- 5.4 mmol/l. Median ICU length of stay (LOS) was 3.0 days (range 2.0-6.0 days, IQR), and hospital LOS was 16 days (range 7-32 days). In 28% of patients insulin treatment was started. Median Apache-II score was 13. 68% of patients were mechanically ventilated. Univariate analysis showed an association with ICU mortality for mean glucose (non-survivors 8.6 +/- 4.3 vs 7.2 +/- 2.4 survivors), maximum glucose (11.7 +/- 5.9 vs 9.6 +/- 5.2, non-survivors vs survivors, respectively), use of insulin (mortality 29 vs 17% in patients not using insulin) and age (61 vs 55.7 years). Gender and a history of diabetes mellitus were not associated with mortality. In a multivariate model, the Apache-II score was the only variable associated with mortality independent of other variables, including mean blood glucose. CONCLUSION: In this retrospective study mean glucose level was not an independent risk factor for mortality in mixed ICU patients. PMID- 16477417 TI - Influence of infusion flow rates on central venous pressure measurements through multi-lumen central venous catheters in intensive care. AB - OBJECTIVE: To study the influence on central venous pressure (CVP), measured at the distal port, of crystalloid infusions administered through the proximal port(s) of a central venous multi-lumen catheter. PATIENTS: Thirty-one intensive care patients. INTERVENTIONS: CVP was measured at the distal port of a multi lumen catheter inserted in the subclavian or internal jugular vein. Using the proximal port(s), saline (0.9%) was infused at rates varying from 2 ml/h to 14,340 ml/h. RESULTS: CVP measured before the infusion and during infusion (after 30 s to 1 min) were not significantly different. Positive pressure ventilation with PEEP (5.6 +/- 2.5 cmH(2)O) and/or norepinephrine infusion (0.25 +/- 0.21 microg kg(-1) min(-1)) did not produce any significant change in CVP during infusion. CONCLUSION: The administration of crystalloids at different flow rates through the proximal port(s) of a multi-lumen catheter placed in the superior vena cava does not affect CVP measurement at the distal port, even in mechanically ventilated patients or patients receiving vasopressors. PMID- 16477416 TI - Time of non-invasive ventilation. AB - Non-invasive ventilation (NIV) is a safe, versatile and effective technique that can avert side effects and complications associated with endotracheal intubation. The success of NIV relies on several factors, including the type and severity of acute respiratory failure, the underlying disease, the location of treatment, and the experience of the team. The time factor is also important. NIV is primarily used to avert the need for endotracheal intubation in patients with early-stage acute respiratory failure and post-extubation respiratory failure. It can also be used as an alternative to invasive ventilation at a more advanced stage of acute respiratory failure or to facilitate the process of weaning from mechanical ventilation. NIV has been used to prevent development of acute respiratory failure or post-extubation respiratory failure. The number of days of NIV and hours of daily use differ, depending on the severity and course of the acute respiratory failure and the timing of application. In this review article, we analyse, compare and discuss the results of studies in which NIV was applied at various times during the evolution of acute respiratory failure. PMID- 16477418 TI - Procalcitonin kinetics in the prognosis of severe community-acquired pneumonia. AB - OBJECTIVES: Procalcitonin (PCT) kinetics is a good prognosis marker in infectious diseases, but few studies of community-acquired pneumonia (CAP) have been performed in intensive care units (ICU). We analyzed the relationship between PCT kinetics and outcome in ICU patients with severe CAP. DESIGN AND SETTING: Prospective observational study in a 16-bed university hospital ICU. PATIENTS: 100 critically ill patients with community-acquired pneumonia. MEASUREMENTS AND RESULTS: Median PCT was 5.2 ng/ml on day 1 and 2.9 ng/ml on day 3. It increased from day 1 to day 3 in nonsurvivors but decreased in survivors. In multivariate analysis four variables were associated with death: invasive ventilation (odds ratio 10-), multilobar involvement (5.6-), LOD score (6.9-), and PCT increase from day 1 to day 3 (4.5-). In intubated patients with a PCT level below 0.95 ng/ml on day 3 the survival rate was 95%. CONCLUSION: Increased PCT from day 1 to day 3 in severe CAP is a poor prognosis factor. A PCT level less than 0.95 ng/ml on day 3 in intubated patients is associated with a favorable outcome. PMID- 16477419 TI - Preservation procedures for arsenic speciation in a stream affected by acid mine drainage in southwestern Spain. AB - A preservation study has been performed for arsenic speciation in surface freshwaters affected by acid mine drainage (AMD), a pollution source characterized by low pH and high metallic content. Two sample preservation procedures described in the literature were attempted using opaque glass containers and refrigeration: i) addition of 0.25 mol L(-1) EDTA to the samples, which maintained the stability of the arsenic species for 3 h; and ii) in situ sample clean-up with a cationic exchange resin, in order to reduce the metallic load, which resulted in a partial co-adsorption of arsenic onto Fe precipitates. A new proposed method was also tried: sample acidification with 6 mol L(-1) HCl followed by in situ clean-up with a cationic exchange resin, which allowed a longer preservation time of at least 48 h. The proposed method was successfully applied to water samples with high arsenic content, taken from the Aguas Agrias Stream (Odiel River Basin, SW Spain), which is severely affected by AMD that originates at the nearby polymetallic sulfide mine of Tharsis. The speciation results obtained by liquid chromatography-hydride generation-atomic fluorescence spectrometry (HPLC-HG-AFS) indicated that during the summer the main arsenic species was As(V) at the hundred microg L(-1) level, followed by DMA (dimethyl arsenic) and As(III) below the ten microg L(-1) level. In winter, As(V) and As(III) increased at least fivefold, whereas the DMA was not detected. PMID- 16477421 TI - Building a professional career. Advice to a new faculty member. PMID- 16477420 TI - Reduction of background interference in the spectrophotometric assay of mevalonate kinase. AB - Mevalonate kinase can be conveniently assayed by coupling to two other reactions and monitoring the consumption of NADH optically at 340 nm. No mevalonate kinase was detected in crude extracts of Saccharomyces cerevisiae strains, however, because of background interference measured in the absence of mevalonate. A strain of S. cerevisiae over-expressing mevalonate kinase was used to establish conditions for reduction of background interference. This method has been successfully applied to S. cerevisiae strains containing a wild type level of mevalonate kinase. PMID- 16477422 TI - Fluorometric determination of sugars using fluorescein-labeled concanavalin A glycogen conjugates. AB - The fluorescence of fluoresceinisothiocyanate-labeled concanavalin A (FITC-Con A) was quenched by forming an FITC-Con A-glycogen conjugate and dequenched upon addition of sugars to the conjugate solution due to disaggregation of the conjugate. However, fluorescence quenching was barely observed upon formation of FITC-Con A-dextran conjugate. The sugar-induced fluorescence response of the FITC Con A-glycogen conjugate depended significantly on the type of sugar: methylated alpha-D: -glucose and alpha-D: -mannose both induced high and rapid responses, while the responses to D: -mannose and D: -glucose were moderate. In contrast, no response was observed in the presence of D: -galactose due to a lack of affinity to Con A. Thus, it is apparent that D: -glucose and other sugars can be detected via the fluorescence of the FITC-Con A-glycogen conjugate. PMID- 16477423 TI - Transient pseudo-isotachophoretic stacking in analysis of plasma for homocysteine by capillary zone electrophoresis. AB - An analytical procedure enabling routine analysis of human plasma for total homocysteine has been developed and validated. The method includes reduction of homocysteine disulfides to thiol with tris 2-carboxyethylphosphine, derivatization of the thiol with 2-chloro-1-methylquinolinium tetrafluoroborate, separation of homocysteine 2-S-quinolinium derivative from those of plasma endogenous and exogenous thiol derivatives by capillary zone electrophoresis, and quantitation with the use of ultraviolet detection based on acetonitrile stacking. Method performance characteristics, for example recovery, calibration, precision, limit of detection, and limit of quantitation, are presented. The procedure was applied to analysis of plasma samples donated by apparently healthy volunteers. PMID- 16477424 TI - Microemulsion electrokinetic chromatographic determination of bufadienolides in toad venom and in traditional Chinese medicine. AB - A microemulsion electrokinetic chromatographic (MEEKC) method has been developed and validated for determination of resibufogenin and cinobufagin in toad venom and in traditional Chinese medicine prepared from the venom. The MEEKC method involved use of sodium dodecyl sulfate (SDS) as surfactant, heptane as organic solvent, and butan-1-ol as co-solvent. To improve the separation, the effect of temperature and running buffer pH were evaluated. The optimized conditions (heptane 0.81% (w/w), SDS 3.31% (w/w), butan-1-ol 6.61% (w/w), and 10 mmol L(-1) sodium tetraborate buffer, pH 9.2, and 298 nm as the detection wavelength) enabled useful and repeatable separation of the analytes. PMID- 16477425 TI - Treatment of impacted lower third ureteral stones with the use of the ureteral access sheath. AB - We present our experience with the use of the ureteral access sheath for the management of small impacted lower third ureteral stones, in comparison with more standard techniques. Ninety-eight consecutive patients, aged 18-73 years (mean 48.5), with small (diameter < or = 10 mm) impacted lower third ureteral stones (< 5 mm in 56, and 5-10 mm in 42 patients) were randomly managed with either a 12/14F coaxial ureteral dilator/sheath and a 7.5F flexible ureteroscope (group A; 48 patients), or with balloon dilatation and the 7.5F flexible ureteroscope (group B; 50 patients). In both groups, stones were grasped and extracted with a basket, and when necessary they were disintegrated with a 1.9F electrohydraulic lithotripsy (EHL) probe. Postoperatively, excretory urography was performed at 1 month and patients were followed-up for 1 year. The mean operative time was 45.5 min in group A, and 58.5 min in group B (P<0.05). EHL was performed in 16 (33.3%) patients of group A, and in 12 (24%) patients of group B. In group B, balloon dilatation was performed in 28 (56%) patients. Ureteral perforation was revealed in 4 (8%) patients of group B. The follow-up imaging tests showed stone-free status in 46 (95.8%) patients of group A and in all (100%) patients of group B. No long-term complications were recorded. Endoscopic management of small impacted lower third ureteral stones with the ureteral access sheath is a quicker and safer procedure, in comparison with the more standard approach, bearing comparable efficacy. PMID- 16477426 TI - Renal colic during sexual intercourse: a unique presentation. AB - The average lifetime risk of renal stones has been reported to be in the range of 5-21%, and the majority of patients have recurrent stones (Tiselius et al. in Eur Urol 40:362-371, 2001). The peak incidence is between the fourth and fifth decades, and therefore generally active and working adults are most affected. Stones are usually formed in a calyx, and become symptomatic if they move to obstruct the upper urinary tract. In the majority of cases, there is no specific action which causes stone movement from a non-obstructing to an obstructing position. We present the first ever case report in the literature of an episode of renal colic during sexual intercourse. The role of percussion therapy and postural drainage are well established following shockwave lithotripsy (SWL) to enhance passage of lower pole stone fragments (Brownlee et al. in J Urol 143:1096, 1990), and it may well be the result of similar principles of motion and body positioning which caused the patient to present in this manner. PMID- 16477427 TI - Stone analysis. AB - The definition and the frequencies of the components of urinary stones are presented on the basis of more than 110,000 analyses. The stone morphologies are described. The methods of stone analysis X-ray diffraction, infrared spectroscopy and polarization microscopy and the most recent developments are presented. The benefits and disadvantages of different methods are compared. The results of ring trials show that the X-ray diffraction method is the best method with regard to correctness. PMID- 16477429 TI - AB-QTL analysis in spring barley: II. Detection of favourable exotic alleles for agronomic traits introgressed from wild barley (H. vulgare ssp. spontaneum). AB - The objective of the present study was to identify favourable exotic Quantitative Trait Locus (QTL) alleles for the improvement of agronomic traits in the BC2DH population S42 derived from a cross between the spring barley cultivar Scarlett and the wild barley accession ISR42-8 (Hordeum vulgare ssp. spontaneum). QTLs were detected as a marker main effect and/or a marker x environment interaction effect (M x E) in a three-factorial ANOVA. Using field data of up to eight environments and genotype data of 98 SSR loci, we detected 86 QTLs for nine agronomic traits. At 60 QTLs the marker main effect, at five QTLs the M x E interaction effect, and at 21 QTLs both the effects were significant. The majority of the M x E interaction effects were due to changes in magnitude and are, therefore, still valuable for marker assisted selection across environments. The exotic alleles improved performance in 31 (36.0%) of 86 QTLs detected for agronomic traits. The exotic alleles had favourable effects on all analysed quantitative traits. These favourable exotic alleles were detected, in particular on the short arm of chromosome 2H and the long arm of chromosome 4H. The exotic allele on 4HL, for example, improved yield by 7.1%. Furthermore, the presence of the exotic allele on 2HS increased the yield component traits ears per m2 and thousand grain weight by 16.4% and 3.2%, respectively. The present study, hence, demonstrated that wild barley does harbour valuable alleles, which can enrich the genetic basis of cultivated barley and improve quantitative agronomic traits. PMID- 16477428 TI - QTL analysis for rice grain length and fine mapping of an identified QTL with stable and major effects. AB - Grain length in rice plays an important role in determining rice appearance, milling, cooking and eating quality. In this study, the genetic basis of grain length was dissected into six main-effect quantitative trait loci (QTLs) and twelve pairs of epistatic QTLs. The stability of these QTLs was evaluated in four environments using an F7 recombinant inbred line (RIL) population derived from the cross between a Japonica variety, Asominori, and an Indica variety, IR24. Moreover, chromosome segment substitution lines (CSSLs) harboring each of the six main-effect QTLs were used to evaluate gene action of QTLs across eight environments. A major QTL denoted as qGL-3a, was found to express stably not only in the isogenic background of Asominori but also in the recombinant background of Asominori and IR24 under multiple environments. The IR24 allele at qGL-3a has a positive effect on grain length. Based on the test of advanced backcross progenies, qGL-3a was dissected as a single Mendelian factor, i.e., long rice grain was controlled by a recessive gene gl-3. High-resolution genetic and physical maps were further constructed for fine mapping gl-3 by using 11 simple sequence repeat (SSR) markers designed using sequence information from seven BAC/PAC clones and a BC4F2 population consisting of 2,068 individuals. Consequently, the gl-3 gene was narrowed down to a candidate genomic region of 87.5 kb long defined by SSR markers RMw357 and RMw353 on chromosome 3, which provides a basis for map-based cloning of this gene and for marker-aided QTL pyramiding in rice quality breeding. PMID- 16477431 TI - [Evaluation of traditional German undergraduate surgical training. An analysis at Heidelberg University]. AB - BACKGROUND: The medical faculty of Heidelberg University implemented a new problem-based clinical curriculum (Heidelberg Curriculum Medicinale, or Heicumed) in 2001. The present study analyses the evaluation data of two student cohorts prior to the introduction of Heicumed. Its aim was to specify problems of the traditional training and to draw conclusions for implementation of a new curriculum. METHODS: The evaluation instrument was the Heidelberg Inventory for the Evaluation of Teaching (HILVE-I). The data were analysed calculating differences in the means between defined groups, with the 13 primary scales of the HILVE I-instrument as dependent variables. RESULTS: Teaching method and subject had no systematic influence on evaluation results. Thus, didactic lecture in orthopedic surgery achieved better results than small group tutorials, while the data on vascular and general surgery showed opposite results. Major factors for success were continuity and didactic training of lecturers and tutors. This is convincingly reflected by the results of the lecture course "Differential diagnosis in general surgery". The good evaluation data on small group tutorials resulted largely from the "participation" and "discussion" scales, which represent interactivity in learning. CONCLUSION: The results of the present study suggest the importance of two major pedagogic ideas: continuity and didactic training of lecturers and tutors. These principles were widely implemented in Heicumed and have contributed to the success of the new curriculum. PMID- 16477430 TI - [Thorax injuries]. AB - Thorax injuries may be divided etiologically into blunt and penetrating types, depending on the nature of the insult. In European practice, the former predominates by far, and in only about 5% of cases thoracotomy provides the necessary thorax drainage. Morbidity in this type of injury typically involves concomitant lung contusion, sometimes with fatal acute respiratory distress syndrome. In these cases, special ventilation forms, optimal reduction of pain, and organ replacement are the decisive therapeutic methods. In contrast, about 80% of penetrating trauma to the thorax require prompt transpleural or trans sternal surgery, depending on the type of injury. Emergency first aid must follow the principle of "scoop and run". Each minute elapsed until emergent thoracotomy can be decisive to survival in these cases, and the fastest possible transport from the place of injury takes priority over time-consuming stabilization. PMID- 16477432 TI - On cloning research, peer review and the possibility of fraud. PMID- 16477433 TI - 99mTc-HYNIC-annexin V SPECT imaging of acute stroke and its response to neuroprotective therapy with anti-Fas ligand antibody. AB - PURPOSE: The first aim of the study was to determine whether (99m)Tc-HYNIC annexin V, a marker of cellular stress and apoptosis, can detect ischemic injury in patients with acute stroke. Secondly, we wished to test radiolabeled annexin's ability to monitor therapy in a rodent model of focal ischemic injury. METHODS: SPECT imaging of patients was performed between 1 and 2 h after intravenous injection of 30 mCi (1,110 MBq) of tracer. Eight MFL4 (anti-FasL) antibody treated (400 microg i.p. days 0 and 3) and 21 control adult male Sprague-Dawley rats underwent small animal SPECT imaging with 5-10 mCi (185-370 MBq) of tracer, 1 and 6 days after a 2-h intraluminal thread occlusion of the left middle cerebral artery. RESULTS: Two patients with acute stroke had regions of multifocal annexin uptake that correlated with sites of restricted diffusion on MRI. Anti-FasL antibody treatment significantly reduced annexin uptake by 92% with a 60% decrease in the number of caspase-8 staining (apoptotic) neurons on day 1. On day 6, treated animals had an 80% reduction in tracer uptake with a 75% decrease in infarct size as compared with controls. Annexin uptake in controls and treated animals (day 6) linearly correlated with infarct size (r (2)=0.603, p=0.0036) and the number of TUNEL-positive (apoptotic) nuclei (r (2)=0.728, p=0.00084). CONCLUSION: Annexin imaging shows foci of increased uptake at sites of ischemic injury in patients with acute stroke. Annexin imaging can assess the effects of therapy for ischemic cerebral injury in rats, suggesting its potential as a non-invasive indicator of drug efficacy in future clinical trials. PMID- 16477434 TI - Structural involvement in substrate recognition of an essential aspartate residue conserved in Mep/Amt and Rh-type ammonium transporters. AB - Ammonium transport proteins belonging to the Mep/Amt/Rh family are spread throughout all domains of life. A conserved aspartate residue plays a key role in the function of Escherichia coli AmtB. Here, we show that the analogous aspartate residue is critical for the transport function of eukaryotic family members as distant as the yeast transporter/sensor Mep2 and the human RhAG and RhCG proteins. In yeast Mep2, replacement of aspartate(186) with asparagine produced an inactive transporter localized at the cell surface, whilst replacement with alanine was accompanied by stacking of the protein in the endoplasmic reticulum. Introduction of an acidic residue, glutamate, produced a partially active protein. A carboxyl group at position 186 of Mep2 therefore appears mandatory for function. Kinetic analysis shows the Mep2(D186E) variant to be particularly affected at the level of substrate affinity, suggesting an involvement of aspartate(186) in ammonium recognition. Our data also put forward that ammonium recognition and/or transport by Mep2 is required for the sensor role played in the development of pseudohyphal growth. Finally, replacement of the conserved aspartate with asparagine in human RhAG and RhCG proteins resulted in the loss of bi-directional transport function. Hence, this aspartate residue might play a preserved functional role in Mep/Amt/Rh proteins. PMID- 16477436 TI - A multi-centre, web-accessible and quality control-checked database of in vivo MR spectra of brain tumour patients. AB - OBJECTIVE: To describe an Internet-accessible database that contains validated in vivo MR spectra and clinical data of brain tumour patients. MATERIALS AND METHODS: All data from patients entering the INTERPRET project (International Network for Pattern Recognition of Tumours Using Magnetic Resonance, ) were stored in a web-accessible database (iDB) and selected using its query functionality. Criteria for selection were that the case had a single voxel (SV) short-echo (20-32 ms) 1.5 T spectrum acquired from a nodular region of the tumour, that the voxel had been positioned in the same region as where subsequent biopsy was obtained, that the short-echo spectrum had not been discarded because of acquisition artefacts or other reasons, and that a histopathological diagnosis was agreed among a committee of neuropathologists. When the spectra were obtained from normal volunteers or were of abscesses or clinically proven metastases, biopsy was not required. RESULTS: A subset of 304 cases (22 normal volunteers and 282 tumour patients) was obtained. These cases were migrated to another similar database (validated-DB). CONCLUSION: The validated-DB complies with ethics regulations and represents the population studied. It is accessible by neuroradiologists willing to use information provided by MRS to help in the non-invasive diagnosis of brain tumours. PMID- 16477435 TI - Normal range of human left ventricular volumes and mass using steady state free precession MRI in the radial long axis orientation. AB - BACKGROUND: The radial long-axis orientation for the measurement of left ventricular (LV) volume and mass has been shown to have advantages over the short axis orientation. Previous work has highlighted the need for technique specific normal ranges. The purpose of this study was therefore to establish normal ranges of LV volume and mass for the radial long-axis orientation. MATERIALS AND METHODS: Forty normal subjects (20 males, average age 32.3, age range 19-58; 20 females, average age 37.4, age range 21-54) were examined utilising a steady state free precession (SSFP) pulse sequence. Two observers analysed the images independently using in-house validated software. RESULTS: The normal ranges for LV end-diastolic volume measurements after adjustment to body surface area (BSA) were 62-120 ml for males and 58-103 ml for females. LV mass indexed to BSA ranged from 50-86 g for males and 36-72 g for females. The normal range for ejection fraction was 49-73% for males and 54-73% for females. CONCLUSION: A gender specific normal range using SSFP in the radial long-axis orientation was established. PMID- 16477437 TI - WNT10B mutations in human obesity. AB - AIMS/HYPOTHESIS: Recent studies suggest that wingless-type MMTV integration site family, member 10B (WNT10B) may play a role in the negative regulation of adipocyte differentiation in vitro and in vivo. In order to determine whether mutations in WNT10B contribute to human obesity, we screened two independent populations of obese subjects for mutations in this gene. SUBJECTS AND METHODS: We studied 96 subjects with severe obesity of early onset (less than 10 years of age) from the UK Genetics of Obesity Study and 115 obese Italian subjects of European origin. RESULTS: One proband with early-onset obesity was found to be heterozygous for a C256Y mutation, which abrogated the ability of WNT10B to activate canonical WNT signalling and block adipogenesis and was not found in 600 control alleles. All relatives of the proband who carried this allele were either overweight or obese. Three other rare missense variants were found in obese probands, but these did not clearly cosegregate with obesity in family studies and one (P301S), which was found in three unrelated subjects with early-onset obesity, had normal functional properties. CONCLUSIONS/INTERPRETATION: These mutations represent the first naturally occurring missense variants of WNT10B. While the pedigree analysis in the case of C256Y WNT10B does not provide definitive proof of a causal link of this variant with obesity, the finding of a non-functioning WNT10B allele in a human family affected by obesity should encourage further study of this gene in other obese populations. PMID- 16477438 TI - Effects of metformin and thiazolidinediones on suppression of hepatic glucose production and stimulation of glucose uptake in type 2 diabetes: a systematic review. AB - AIMS/HYPOTHESIS: Insulin resistance, which manifests itself as endogenous glucose overproduction and reduced insulin-mediated glucose uptake, is a core defect in type 2 diabetes. Metformin and the peroxisome proliferator-activated receptor gamma agonists, the thiazolidinediones (TZDs), both lower glucose, although their mechanism of action is still subject to debate. This review analyses the evidence relevant to these mechanisms in vivo. MATERIALS AND METHODS: A systematic search of MEDLINE identified a total of 42 clinical studies that investigated the effects of TZDs (n=23) and/or metformin (n=19) on endogenous glucose production (using tracer glucose techniques) and peripheral glucose disposal (using the euglycaemic-hyperinsulinaemic clamp) in patients with type 2 diabetes (n=549). The original variables assessed were converted into standardised units and their mean group values were listed separately for open and placebo-controlled studies. Statistical analysis was scarried out, treating mean group values as individual values and comparing results (both as absolute values and percentage changes from baseline) across study categories (open vs placebo-controlled, TZDs vs metformin). RESULTS: Both TZDs and metformin enhance insulin suppression of endogenous glucose production and fasting plasma glucose clearance. TZDs, but not metformin, also improve insulin-mediated glucose uptake at all insulin levels. CONCLUSIONS/INTERPRETATION: In patients with type 2 diabetes, metformin improves fasting hepatic insulin sensitivity and glucose clearance; TZDs improve fasting hepatic insulin sensitivity and glucose clearance, and potentiate glucose disposal under insulinised conditions. PMID- 16477439 TI - Vascular and metabolic effects of methacholine in relation to insulin action in muscle. AB - AIMS/HYPOTHESIS: Methacholine (MC) is a nitric oxide vasodilator, but unlike other vasodilators, it potentiates insulin-mediated glucose uptake by muscle. The present study aimed to resolve whether this action was the result of a vascular effect of MC leading to increased muscle perfusion or a direct effect of MC on the myocytes. We hypothesise that vascular-mediated insulin-stimulated glucose uptake responses to MC occur at lower doses than direct myocyte MC-mediated increases in glucose uptake. METHODS: The vascular and metabolic effects of this vasodilator were examined in rats in vivo using a novel local infusion technique, and in the pump-perfused rat hindlimb under conditions of constant flow. RESULTS: Local infusion of low-dose MC (0.3 micromol/l) into the epigastric artery of one leg (test) in vivo markedly increased femoral blood flow and decreased vascular resistance, without effects in the contra-lateral leg. Capillary recruitment, but not glucose uptake, was increased in the test leg. All increases caused by MC were confined to the test leg and blocked by local infusion into the test leg of N-nitro-L-arginine methyl ester (L-NAME), but not by infusion of N-nitro-D arginine methyl ester (D-NAME). In the constant-flow pump-perfused rat hindlimb, infusion of 0.6 micromol/l MC vasodilated the pre-constriction effected by 70 nmol/l noradrenaline or 300 nmol/l serotonin, and this was blocked by 10 micromol/l L-NAME. 2-Deoxyglucose in muscle was increased by 30 micromol/l MC (p<0.05), but was unaffected by 3 micromol/l MC. All increases in 2-deoxyglucose uptake by 30 micromol/l MC were blocked by 10 micromol/l L-NAME. CONCLUSIONS/INTERPRETATION: MC has dose-dependent effects both on the vasculature and on muscle metabolism. At low dose (0.3-3 micromol/l), MC is a potent vasodilator in muscle, both in vivo and in vitro, without metabolic effects; at higher doses (> or =30 micromol/l) MC has a direct metabolic effect leading to increased glucose uptake. Both the vascular and metabolic effects are sensitive to L-NAME. The low-dose enhancement of insulin action in vivo by MC, which has been reported previously, thus seems to be attributable to vascular effects. PMID- 16477440 TI - Clinical presentation of posterior fossa epidermoid cysts. AB - The aim of this study was to investigate the audiovestibular deficits in those with posterior fossa epidermoid cyst including cerebellopontine angle in two patients and cerebellum in one patient. Prior to operation, all three patients showed bilateral gaze nystagmus. Audiometry revealed mild hearing loss in two patients, and caloric test displayed canal paresis in two patients. After operation, subsidence of gaze nystagmus and recovery of caloric responses were disclosed in all patients. Two patients had both hearing and vestibular evoked myogenic potential (VEMP) results recovered to normal. In contrast, the only one who underwent craniotomy twice displayed bilateral mild hearing loss and delayed VEMPs, possibly due to operation sequela. Hence, bilateral gaze nystagmus may present as an initial sign for posterior fossa epidermoid cyst. Audiovestibular deficits in cases of epidermoid cyst are attributable to compression neuropathy, which may resolve and return to normal responses after surgery. PMID- 16477442 TI - Induction of apoptosis in leukemia cell lines by Linum persicum and Euphorbia cheiradenia. AB - PURPOSE: In the present study two medicinal herbs Linum persicum and Euphorbia cheiradenia that are native to Iran were tested for their possible anticancer effect and induction of apoptosis on human tumor cell lines including leukemia cell lines. METHODS: The effect of methanolic extracts of the herbs on the inhibition of cell proliferation was assessed by MTT colorimetric assay. K562 and Jurkat cell lines treated with the extracts were analyzed for the induction of apoptosis by flow cytometry using propidium iodide staining. DNA fragmentation analysis was performed. RESULTS: Various concentrations of L. persicum and E. cheiradenia showed inhibitory effects on different cell lines. Two leukemic lines including K562 and Jurkat were the most sensitive cells for L. persicum with IC50 of 0.1 and 10 mug/ml, respectively. In the cultures of tumor cell lines treated with E. cheiradenia, the main inhibitory effects was for Jurkat cells with IC50 of 12.5 microg/ml. Results indicated a dose-dependent accumulation of cells in the sub-G1 phase. Study of internucleosomal DNA fragmentation showed a typical DNA laddering in agarose gels for both extracts. CONCLUSION: The present study showed cytotoxic activity of both herbs on tumor cell lines and suggests that this effect may in part be due to the induction of apoptosis in leukemic cells. PMID- 16477443 TI - Production of poly(L-lactide)-degrading enzyme by Amycolatopsis orientalis for biological recycling of poly(L-lactide). AB - Efficient production of poly(L-lactide)(PLA)-degrading enzyme was achieved by addition of 0.1% (w/v) silk fibroin powder into a liquid culture medium of an actinomycete, Amycolatopsis orientalis, without other complex nitrogen sources, such as yeast extract and peptone. Scaled-up production of the enzyme in a 5-l jar fermenter showed the possibility of producing this enzyme on an industrial scale at low production cost. The extracellular PLA-degrading enzyme showed potent degrading activity, which is effective for biological recycling of PLA, i.e., 2,000 mg/l of PLA powder was completely degraded within 8 h at 40 degrees C using 20 mg/l purified enzyme. An optically active L-lactic acid with 600 mg/l was obtained as degradation product of PLA without undesirable racemization. PMID- 16477441 TI - Relationship between hepatitis C virus (HCV) and insulin resistance, endothelial perturbation, and platelet activation in HIV-HCV-coinfected patients under highly active antiretroviral treatment. AB - Insulin resistance is associated with highly active antiretroviral therapy in HIV infected patients, and the risk of developing insulin resistance is increased in hepatitis C virus (HCV)-infected patients. The aim of the present study was to determine whether hepatitis C virus infection constitutes an additional risk factor for insulin resistance or other prothrombotic conditions in HIV-HCV coinfected patients under highly active antiretroviral therapy. One hundred eighteen HIV-infected patients were studied: 50 who had no history of anti-HIV treatment and 68 who were receiving therapy with highly active antiretroviral treatment. The treatment-naive group consisted of 35 HCV-negative subjects and 15 HCV-positive ones. Within the treated group, 50 patients were HCV negative and 18 were HCV positive. For each patient, the lipid profile was determined and the following values measured: glucose, soluble P-selectin (as a marker of platelet activation), soluble thrombomodulin, von Willebrand factor and soluble vascular cell adhesion molecule-1 (as endothelial markers), and insulin resistance. No significant difference (p>0.05) for any variable was found among subjects with or without HCV coinfection in the treatment-naive group. Among patients under highly active antiretroviral therapy, however, those with HCV coinfection showed higher values (p<0.05) for insulin resistance (homeostasis model assessment value: 2.65 vs. 1.79), glucose (93 vs. 86 mg/dl), endothelial markers (von Willebrand factor, 204 vs. 123%; soluble vascular cell adhesion molecule-1, 650 vs. 482 ng/ml), and platelet activation marker (soluble P-selectin, 78 vs. 51 ng/ml) in parallel with lower CD4+ cells counts (289 vs. 402 cells/mm3) and higher HIV-1 viral loads (305 vs. 50 copies/ml) compared to patients without HCV coinfection. Glucose, soluble P-selectin, and von Willebrand factor were independently related to HCV infection. The presence of HCV coinfection during HIV treatment was closely related to higher values of insulin resistance, to activated platelets, and to endothelial perturbation in parallel with lower CD4+ cell counts and higher HIV-1 viral loads compared to patients without HCV coinfection. On the basis of these results, it may be preferable to treat HCV infection prior to initiating treatment for HIV infection in HIV-HCV-coinfected patients. PMID- 16477444 TI - Torque and surface mechanomyogram parallel reduction during fatiguing stimulation in human muscles. AB - The purpose of the study was to verify, by means of torque and mechanomyogram (MMG) compared analysis, the validity of MMG as a tool to investigate the contractile changes due to localized muscular fatigue induced by stimulation protocols usually employed for sport training and rehabilitation programs. Ten healthy sedentary subjects participated in the study. Torque produced by the dominant biceps brachii (BB) and vastus lateralis (VL) during transcutaneous stimulated contractions has been recorded by a load cell strapped to the subjects' wrist and distal one-third of the tibia, respectively. MMG was detected over the muscle bellies during a monopolar supramaximal stimulation of the main motor point. After potentiation, the fatiguing stimulation was administered. It consisted of 50 cycles, with 2 s of 50 Hz and 25 s of 2 Hz. Averaged normalized values of peak torque (pT) and MMG peak-to-peak (MMG-pp) of the subjects group decreased from their initial 100% values to 55% (pT) and 60% (MMG-pp) for BB and to 43% (pT) and 47% (MMG-pp) for VL. The pT% and MMG-pp% changes throughout the stimulation protocol presented high correlation (BB: R=0.95, P<0.001; VL: R=0.94, P<0.001). This correlation suggests that MMG could be used to follow muscle mechanical fatigue development when torque output is not or hardly detectable such as during electrical stimulation programs employed for sport training or rehabilitation protocols. PMID- 16477445 TI - Effect of training and detraining on catecholamine responses to sprint exercise in adolescent girls. AB - Training is well known to influence catecholamine responses to exercise. In women, this training effect is still not well characterized and has been studied mostly in adults. Hence, we investigated in this longitudinal study, the effects of a 6-month sprint training program followed by 5 months of detraining on plasma catecholamine responses to a sprint exercise in young female subjects. Twelve healthy adolescent girls [training group (TG), n=6; control group (CG), n=6] took part in our study. TG participated in 6 months of supervised sprint training program (3 days/week) and has no training past whereas, CG continued with it's normal activity. A 6s-sprint test was performed on a cycle ergometer before training (P1) and after training (P2) in both the groups. TG only realized a 6s sprint test after 5 months of detraining (P3). Blood lactate concentrations (La) as well as plasma adrenaline (A) and noradrenaline (NA) concentrations were measured at rest, immediately after the warm-up and the 6s-sprint and during recovery. Peak power W peak), expressed both in absolute and relative values, were significantly increased in TG in P2 (P<0.01) but did not change in CG. After the sprint-training period, the warm-up and the 6s-sprint induced plasma A increase and the maximal A concentrations were significantly higher than in P1 and P3 for TG only (P<0.05). Plasma A did not change in CG after 6 months. In P3, W peak and maximal lactate concentrations ([La]max) were significantly greater compared to P1 and P2 in TG (P<0.05). In CG, [La]max were significantly increased in P2 (P<0.05). The present study demonstrates that 6 months of sprint training in adolescent girls induce both an increase in performances and in A responses to sprint exercise. This adrenergic adaptation disappears after 5 months of detraining whereas the gain in performance is maintained. These new data may lead to practical considerations. PMID- 16477446 TI - Influence of the contact time on coupling time and a simple method to measure coupling time. AB - The enhancement of performance in stretch shortening cycle (SSC) exercises has been attributed to the recoil of elastic energy stored during the stretching phase and depends on the duration of the coupling time (T(coupling)) i.e., the duration of the isometric phase occurring between the stretch and the shortening of the muscle. However, instead of T(coupling), the contact time (T(contact))- i.e., the sum of T(coupling) plus the duration of the stretching and shortening phases that precede and follow T(coupling)--is more easily and often measured. The aim of this study was to investigate the T(coupling) changes within a large range of T(contact), in order to propose a possible relationship between T(coupling) and T(contact), thus allowing the accurate measurement of T(coupling )only from a tachometer and force data obtained classically in vertical jumps, jumps on sledge apparatus and running on force treadmills. Eleven subjects performed SSC exercises on a sledge apparatus with a large range of T(contact) (400, 700, 1,000, 1,500, 2,000 and 2,500 ms). The T(coupling) and T(contact) values were measured individually, from force platform recordings and the velocity of the carriage seat obtained by a tachometer. For the longest T(contact) (i.e., from 850 to 2,500 ms), we observed a significant linear relationship between T(contact) and T(coupling). This transition between T(contact) shorter or longer than about 850 ms seems to be important and to correspond to T(coupling) close to 300 ms. This limit observed in the present study could be explained physiologically due to a possible modification of the cross-bridges formation. PMID- 16477447 TI - Intraoperative measurement of pharynx/esophagus retraction during anterior cervical surgery. Part II: perfusion. AB - Early postoperative dysphagia after anterior cervical surgery is a well-known phenomenon with so far unknown etiology. We hypothesised that direct pressure induced by the medial retractor blade on pharynx/esophagus mucosal wall leads to local ischemia. Subsequently postoperative hyperemia and swelling of the pharynx/esophagus may result in swallowing disturbance. To prove the hypothesis local blood flow inside the pharynx/esophagus wall during anterior cervical surgery was measured using a laser Doppler (LD) perfusion monitor unit. Fifteen patients underwent standard anterior cervical discectomy and fusion (ACDF). The LD probe was placed underneath the medial retractor blade in order to gain information at the maximum point of pressure applied onto the pharynx/esophagus wall. Local perfusion was measured prior to retractor opening (5 min), during spreading of the retractor and after its closure (5 min). Perfusion was measured semiquantitatively in perfusion units (PU). Local perfusion ranged from 30 to 210 PU (mean 107) prior to retractor opening, from 7 to 60 PU (mean 30) with open retractor and from 15 to 280 PU (mean 117) after retractor closure. In all 15 patients the open retractor led to hypoperfusion ranging from 21 to 93% compared to the baseline level. In seven patients a reactive hyperemia at the end of the procedure was detected (32-89% compared to baseline level). In four patients after hypoperfusion during spreading of the retractor the baseline levels were reached again and in four patients perfusion remained diminished even after retractor closure. To best of our knowledge, this is the first report on intraoperative measurement of local perfusion of the pharynx/esophagus wall during anterior cervical surgery. Diminished local perfusion was observed in all patients during spreading of the retractor and post-procedure hyperemia was recorded in 46% of the patients. The local ischemia of the pharynx/esophagus wall may be a crucial step in the development of postoperative dysphagia. PMID- 16477448 TI - Intraoperative measurement of pharynx/esophagus retraction during anterior cervical surgery. Part I: pressure. AB - Temporary dysphagia after anterior cervical discectomy (ACD) is common. However, its mechanism is poorly understood. Pressure induced by retractor blades onto pharynx/esophagus were measured intraoperatively in order to gain more information regarding traumatization of the pharynx/esophagus wall. Thirty-one patients underwent anterior cervical discectomy and fusion (ACDF) for degenerative disc disease. An online pressure transducer was applied to the rear side of the medial retractor blade (epi-esophageal-pressure, epi-P) and a cylindric, inflatable transducer was preoperatively inserted into the pharynx/esophagus under fluoroscopic guidance at the level to be operated on (endo-esophageal-pressure, endo-P). Pressure values were recorded continuously during the operation. Mean arterial pressure (MAP) and endotracheal cuff pressure (ETCP) were recorded additionally. An in vitro model was developed in order to analyze the impact of the retractor blade design onto the epi-esophageal pressure. Mean epi-P before and following adequate retractor opening for exposure of the disc space was 58.3 and 92.7 mmHg. Thirty, 60 and 90 min later the epi-P decreased to 79, 70 and 66%, respectively. Mean basal endo-P was 9.8 mmHg and increased to 20.6 mmHg after retractor placement. Thirty, 60 and 90 min later the endo-P decreased to 80, 71 and 62%, respectively. The mean MAP was 76 mmHg and the ECTP was adjusted to 25 mmHg during the procedures. In the in vitro model retraction pressure correlated inversely with the contact area between visceral wall and retractor blade. During ACDF the retraction pressure onto the pharyngeal/esophageal wall exceeds MAP and even more the mucosal perfusion pressure of 25 mmHg. Over time the pharynx/esophageal wall adapts to the applied pressure induced by the retractor blade. The contact area between them influences the retraction pressure. PMID- 16477449 TI - Variability of spinal instrumentation configurations in adolescent idiopathic scoliosis. AB - Surgical instrumentation for the correction of adolescent idiopathic scoliosis (AIS) is a complex procedure involving many difficult decisions (i.e. spinal segment to instrument, type/location/number of hooks or screws, rod diameter/length/shape, implant attachment order, amount of rod rotation, etc.). Recent advances in instrumentation technology have brought a large increase in the number of options. Despite numerous clinical publications, there is still no consensus on the optimal surgical plan for each curve type. The objective of this study was to document and analyse instrumentation configuration and strategy variability. Five females (12-19 years) with AIS and an indication for posterior surgical instrumentation and fusion were selected. Curve patterns were as follows: two right thoracic (Cobb: 34 degrees, 52 degrees), two right thoracic and left lumbar (Cobb T/L: 57 degrees/45 degrees, 72 degrees/70 degrees) and 1 left thoraco-lumbar (Cobb: 64 degrees). The pre-operative standing postero anterior and lateral radiographs, supine side bending radiographs, a three dimensional (3D) reconstruction of the spine, pertinent 3D measurements as well as clinical information such as age and gender of each patient were submitted to six experienced independent spinal deformity surgeons, who were asked to provide their preferred surgical planning using a posterior spinal approach. The following data were recorded using the graphical user interface of a spine surgery simulator (6x5 cases): implant types, vertebral level, position and 3D orientation of implants, anterior release levels, rod diameter and shape, attachment sequence, rod rotation (angle, direction), adjustments (screw rotation, contraction/distraction), etc. Overall, the number of implants used ranged from 11 to 26 per patient (average 16; SD +/-4). Of these, 45% were mono axial screws, 31% multi-axial screws and 24% hooks. At one extremity of the spectrum, one surgeon used only mono-axial screws, while at the other, another surgeon used 81% hooks. The selected superior- and inferior-instrumented vertebrae varied up to six and five levels, respectively (STD 1.2 and 1.5). A top to-bottom attachment sequence was selected in 61% of the cases, a bottom-up in 29% and an alternate order in 11%. The rod rotation maneuver of the first rod varied from 0 degrees (no rotation) to 140 degrees, with a median at 90 degrees. In conclusion, a large variability of instrumentation strategy in AIS was documented within a small experienced group of spinal deformity surgeons. The exact cause of this large variability is unclear but warrants further investigation with multicenter outcome studies as well as experimental and computer simulation studies. We hypothesize that this variability may be attributed to different objectives for correction, to surgeon's personal preferences based on their previous experience, to the known inter-observer variability of current classification systems and to the current lack of clearly defined strategies or rational rules based on the validated biomechanical studies with modern multi-segmental instrumentation systems. PMID- 16477450 TI - Comment on "Os odontoideum with bipartite atlas and segmental instability: a case report" (M. Osti et al.). PMID- 16477451 TI - Comments on "Discriminative validity and responsiveness of the Oswestry disability index among Japanese outpatients with lumbar conditions". PMID- 16477452 TI - Discriminative validity and responsiveness of the Oswestry Disability Index among Japanese outpatients with lumbar conditions. AB - The Oswestry Disability Index (ODI) is one of the most used assessment scales for patients with spine conditions, and translations into several languages have already been available. However, the scale's discriminative validity and responsiveness to the clinical change was somewhat understudied in these translated versions of the ODI. In this study, we independently developed a Japanese version of the ODI, and tested its discriminative and responsive performances among outpatients with various spinal conditions. We recruited 167 outpatients from seven participating clinics, and concurrently measured the translated ODI and MOS Short Form 36 (SF36) as a reference scale. We also obtained from medical records clinical information such as diagnoses, the past history of surgery, and existence of subjective symptoms and clinical signs. For testing discriminative validity, scores were compared by the number of symptoms and signs, with the trend test. Receiver operating characteristics (ROC) analysis was also conducted to compare ODI and SF36 in their performance to discriminate the existence of signs/symptoms, by chi-square test on the area under ROC curve (AUC). For 35 patients (17 clinically stable, 18 undergoing surgery and clinically significantly changed), the two scales were repeatedly administered after 3-6 months to compare responsiveness by using ROC analysis. The translated ODI and the SF36 Physical Function (PF) subscale showed a significant trend increase as the numbers of symptoms/signs increased. They also showed comparable performance in discriminating the existence of signs/symptoms (AUC=0.70-0.76 for ODI, 0.69-0.70 for SF36 PF, P=0.15-0.81), and clinical status change over time (AUC=0.82 for ODI, 0.72 for SF36 PF, P=0.31). Our results showed that the translated Japanese ODI showed fair discriminative validity and responsiveness as the original English scale showed. PMID- 16477454 TI - Localized nodular pigmented villonodular synovitis of the upper ankle joint- diagnosis and treatment. AB - This paper introduces a case of local pigmented villonodular synovits (PVNS) of the upper ankle joint in a 37-year old patient. PVNS is a neoplasia of the synovial membrane. Two different entities of PVNS are known: generalized diffuse and local nodular. They differ in their degree of destruction and growth configuration, which is crucial for prognosis and operative treatment. The most common location of the local nodular form is the knee joint, followed by the finger joints. Occurrence in the ankle joint is not common but should be considered if clinical findings are present. MRI is the method of choice for diagnosis and assessment of the bone situation. Treatment consists of radical excision of the neoplasia. Radiosynoviorthesis is recommended as a post-operative treatment to increase the probability of a total removal of persisting PVNS cells. The rate of recurrence seems to be between 8 and 46%. PMID- 16477453 TI - [Effectiveness and safety of radium chloride in the treatment of ankylosing spondylitis. Results of an observational study]. AB - After radium chloride was approved for the therapy of ankylosing spondylitis (AS) in Germany on October 23, 2000, an open, non-comparative prospective observational study was planed. The assessment involved 308 patients with the diagnosis of AS and radiological evidence of spinal involvement who were treated with at least one injection of radium chloride. 278 of these patients received the full cycle of 10 injections (a total of 10 MBq radium-224) one injection one per week for a period of 10 successive weeks. Validated outcomes were measured before therapy and six months after therapy according to the recommendations of the "assessment in ankylosing spondylitis working group"(ASAS). The primary endpoint was disease activity assessed using the BASDAI ("Bath Ankylosing Spondylitis Disease Activity Index"). Secondary endpoints included the BASFI ("Bath Ankylosing Spondylitis Functional Index"), as well as self-reported pain and global patient ratings. Patients showed significant improvement in all of the parameters assessed (BASDAI, BASFI, pain, global patient rating) at 6-months post treatment. At this time, the mean BASDAI and BASFI scores improved from 5.5 to 3.5 and from 5.6 to 3.7, respectively. Pain decreased from a mean of 7.3 to 4.1, and disease activity (as rated by patients) also decreased from 7.2 to 4.1 (p < 0.001 for all of them). Drug tolerance was good. Six adverse events were documented, none of which was serious and none led to discontinuation of treatment. According to the data provided here medical treatment with radium chloride seems to be effective and safe during an observation period of 6 months. Nevertheless, a randomized placebo-controlled study should be performed in the near future in order to safeguard the level of evidence. PMID- 16477455 TI - C-type natriuretic peptide and its receptor are downregulated in pulmonary epithelium following birth. AB - C-type natriuretic peptide (CNP) is a member of the natriuretic peptide family and acts through the membrane bound guanylyl cyclase linked natriuretic peptide receptor B (NPR-B) to increase intracellular cGMP. Activation of the CNP/NPR-B pathway in pulmonary epithelium has been linked to the inhibition of amiloride sensitive sodium absorption and to the stimulation of the cystic fibrosis transmembrane conductance regulator (CFTR). Given the importance of ion movement across the pulmonary epithelium of the fetal and newborn lung, we sought to examine the expression of CNP and NPR-B in pulmonary epithelium of the developing fetal lamb and following the transition to air breathing. Lambs were sacrificed at 100 and 136 days of gestation and at 3 days, and 4 weeks after full term delivery. Lung sections were immunostained for CNP and NPR-B. At 100 days of gestation, staining for CNP and NPR-B was absent within all pulmonary epithelium. At 136 days of gestation, prominent staining for both CNP and NPR-B was seen within alveolar type II cells, non-ciliated cells of the distal airways (Clara cells), and ciliated epithelium of the upper airways. At both 3 days and 4 weeks following birth, staining for CNP and NPR-B was absent in alveolar type II cells, ciliated bronchial epithelium and was markedly reduced in Clara cells. The presence of CNP and NPR-B within the pulmonary epithelium in the nearterm fetal period and its rapid downregulation following birth suggests that CNP may contribute to the maintenance of the fluid-filled lung through the regulation of trans-epithelial ion flux. PMID- 16477456 TI - [The critical role of research evidence in the framework of health technology assessment]. AB - In spite of their different aims, evidence-based medicine (EBM) and health technology assessment (HTA) to some extent use similar methodologies for the assessment of a technology's efficacy. The decision to include only a certain quality of studies in a HTA report usually follows the hierarchy of evidence with different levels of evidence. Problems arise if study evidence is completely lacking or if it is extrapolated or transferred to other healthcare settings. As subjectivity is inevitable when critically appraising the evidence or combining it with other sources of information, HTA decisions should be presented with maximum transparency. Since physicians usually have only limited knowledge of research methodology and read only an average of 20 articles per year, a cooperative approach is required when appraising the evidence for HTA. PMID- 16477457 TI - [Epidemiological studies in the HTA evaluation process]. AB - Results of epidemiological studies should be considered as part of the available evidence when evaluating drug treatment benefits in health technology assessment (HTA). Pharmaco-epidemiological databases can provide a broader understanding of the effectiveness of drugs in populations that are frequently underrepresented in clinical trials. Such databases are also useful to investigate drug safety with regard to socio-demographic and medical care-related indicators and hereby contribute to an optimal and targeted pharmacological therapy. Using examples from pharmaco-epidemiological asthma studies, the present article discusses associated difficulties in interpreting database results against the background of various sources of bias and proposes possibilities for integrating observational data into the HTA evaluation process. Researchers are challenged to engage in considerable efforts to develop a standardized inventory of epidemiological methods, e.g. for the pooled analysis of epidemiological data. PMID- 16477458 TI - Bilayer lipid composition modulates the activity of dermaseptins, polycationic antimicrobial peptides. AB - The primary targets of defense peptides are plasma membranes, and the induced irreversible depolarization is sufficient to exert antimicrobial activity although secondary modes of action might be at work. Channels or pores underlying membrane permeabilization are usually quite large with single-channel conductances two orders of magnitude higher than those exhibited by physiological channels involved, e.g., in excitability. Accordingly, the ion specificity and selectivity are quite low. Whereas, e.g., peptaibols favor cation transport, polycationic or basic peptides tend to form anion-specific pores. With dermaseptin B2, a 33 residue long and mostly alpha-helical peptide isolated from the skin of the South American frog Phyllomedusa bicolor, we found that the ion specificity of its pores induced in bilayers is modulated by phospholipid-charged headgroups. This suggests mixed lipid-peptide pore lining instead of the more classical barrel-stave model. Macroscopic conductance is nearly voltage independent, and concentration dependence suggests that the pores are mainly formed by dermaseptin tetramers. The two most probable single-channel events are well resolved at 200 and 500 pS (in 150 mM NaCl) with occasional other equally spaced higher or lower levels. In contrast to previous molecular dynamics previsions, this study demonstrates that dermaseptins are able to form pores, although a related analog (B6) failed to induce any significant conductance. Finally, the model of the pore we present accounts for phospholipid headgroups intercalated between peptide helices lining the pore and for one of the most probable single-channel conductance. PMID- 16477459 TI - Effect of meal size on postprandial metabolic response in Chinese catfish (Silurus asotus Linnaeus). AB - The effect of relative meal size (0.5-24% body mass) on specific dynamic action (SDA) was assessed in Chinese catfish (Silurus asotus Linnaeus) (30.90+/-1.30 g) at 25.0 degrees C; the cutlets of freshly killed loach without viscera, head and tail were used as a test meal. There was no significant difference in either SDA duration or peak oxygen consumption (VO2) among low meal size ranges. But both increased linearly as meal size increased from 2 to 24% without reaching a plateau. Factorial metabolic scope was 5.92 in fish fed with 24% body mass, the highest documented feeding metabolic scope value in fish till now. The Peak VO2 of satiated meal size groups (175.85+/-10.55 mg O2 h(-1)) was above 80% of maximum metabolic rate during locomotion recovery process (215.48+/-7.07 mg O2 h( 1)). The relationship between energy expended on SDA (E) and energy ingested (I) was described as: E=0.0000432I(2)+0.140I+2.12. The lowest value of SDA coefficient appeared at 2% body mass group. PMID- 16477461 TI - Health care and patients' migration across Spanish regions. AB - This paper analyses patient mobility across Spanish regions. A model of patient migration is specified and estimated using panel observations covering mobility and other main regional quality indicators over the period 1996-1999. Empirical results show that in Spain income and supply variables determines the quality of the service offered,, and that there is quality-driven mobility. PMID- 16477460 TI - Comparative anatomy of gall development on Gypsophila paniculata induced by bacteria with different mechanisms of pathogenicity. AB - Galls induced on Gypsophila paniculata by Pantoea agglomerans pv. gypsophilae (Pag) and Agrobacterium tumefaciens (At), bacteria with different mechanisms of pathogenicity, were compared morphologically and anatomically. The pathogenicity of Pag is dependent on the presence of an indigenous plasmid that harbors hrp gene cluster, genes encoding Hop virulence proteins and biosynthetic genes for auxin (IAA) and cytokinins (CKs), whereas that of At involves host transformation. The Pag-induced gall was rough, brittle and exhibited limited growth, in contrast to the smooth, firm appearance and continuous growth of the At-induced gall. Anatomical analysis revealed the presence of cells with enlarged nuclei and multiple nucleoli, giant cells and suberin deposition in Pag that were absent from At-induced galls. Although circular vessels were observed in both gall types, they were more numerous and the vascular system was more organized in At. An aerenchymal tissue was observed in the upper part of the galls. Ethylene emission from Pag galls, recorded 6 days after inoculation, was eight times as great as that from non-infected controls. In contrast, a significant decrease in ethylene production was observed in Gypsophila cuttings infected with Pag mutants deficient in IAA and CK production. The results presented are best accounted for by the two pathogens having distinct pathogenicity mechanisms that lead to their differential recognition by the host as non-self (Pag) and self (At). PMID- 16477462 TI - [Ultrastructural changes in human nasal mucosa in rhinitis medicamentosa]. AB - BACKGROUND: Long-term abuse of decongestive nasal drops causes rhinitis medicamentosa due to cytotoxic and ciliary-toxic effects. Nasal obstruction is caused by rebound swelling when the decongestive effect has disappeared. The patient starts using nasal drops more frequently as a result of tachyphylaxis. PATIENTS AND METHODS: Tissue samples from human inferior turbinates from 22 patients who had overused decongestive nose drops were taken during nasal surgery and preserved in phosphate-buffered paraformaldehyde or glutaraldehyde. Ultrathin sections were cut. The samples were dehydrated and embedded in Araldit. The findings were photo-documented using a light- and transmission electron microscope. Biopsies from ten patients without chronic inflammation of the nasal mucosa were used as controls. RESULTS: The electron microscopic investigations revealed epithelium showing severe damage corresponding to regions with hyperplastic and metaplastic changes. Loss of ciliated cells was observed. Under a thick basal membrane, ultrastructural changes to the endothelial lining, such as openings and rupture of the basal lamina, were detected. Prominent endothelial cells were conspicuous. CONCLUSIONS: Rhinitis medicamentosa is a drug-induced injury to human nasal mucosa associated with the prolonged abuse of topical nasal decongestants. Loss and destruction of ciliated epithelial cells are the morphological correlation to the disturbance of mucociliary clearance. Endothelial cells of capillaries, in particular, revealed ultrastructural changes indicative of increased permeability with consecutive interstitial edema. PMID- 16477465 TI - [Toxic anosmia and rhinitis due to occupational solvent exposure]. AB - A patient's case report documents the development of anosmia and rhinitis caused by occupational exposure to organic solvents, including tetrahydrofuran, cyclohexanone, acetone and other ketones. Physicians should inquire about possible mucosal irritation or prenarcotic symptoms during exposure. If an occupational disease is assumed, notification must be sent to the Institution for Statutory Accident Insurance and Prevention (Berufsgenossenschaft). Some diseases, for example olfactory disorders due to chemical mixtures, are not included in the German list of occupational diseases. In this case, a special law (Sozialgesetzbuch VII) regulates compensation. PMID- 16477463 TI - [Dysphagia and dysarthria due to the long-term growth of a tongue base tumor]. PMID- 16477466 TI - [Solitary fibrous tumor of the nose and paranasal sinuses]. AB - OBJECTIVE: The solitary fibrous tumor is an uncommon, benign lesion with a mesenchymal origin which arises most commonly from the pleura and, in extremely rare cases, from the mucosa of the nose and paranasal sinuses. PATIENT AND METHODS: We describe a case of solitary fibrous tumor in the nasal cavity with an extension into both ethmoid sinuses and destruction of the nasal septum in a 64 year-old woman presenting with nasal obstruction and frontal headache. In the histopathologic examination and by immunohistochemistry, the tumor was composed of spindle cells and nodular collagen, and was confirmed as a solitary fibrous tumor. It was completely removed using an endonasal approach with the patient being free of any complaints and without evidence of disease 2 years after surgery. CONCLUSIONS: Solitary fibrous tumor should be included in diagnostic considerations in patients with sinonasal masses, especially in cases with the appearance of spindle cells. Diagnostic procedures, clinicopathological features, therapeutic options and prognosis of the such tumors are discussed. PMID- 16477467 TI - Successful treatment for intractable chylous ascites in a child using a peritoneovenous shunt. AB - Intractable post-operative chylous ascites had been managed successfully using a peritoneovenous shunt (PVshunt). A 4-year-old girl with neuroblastoma originated from the right adrenal gland was admitted to our hospital. Following the preoperative chemotherapy, tumor resection, and lymph node dissection of the abdominal paraaortic region were carried out. Post-operative radiation therapy 9.6 gray to the tumor bed and to the paraaortic region and a high dose chemotherapy supported by auto bone marrow transplantation were completed. Three months later some enlarged lymph nodes along the duodeno-hepatic ligament were detected and these had gradually increased in size. Lymph node dissection along the hepatic artery and the abdominal aorta was carried out. Pathological examination of the specimen showed reactive lymph node swelling. Chylous ascites developed several days after surgery. Despite the medium-chain triglycerides meal or total parental nutrition, the ascites persisted for more than 80 days. Multiple paracenteses were mandatory. A PV shunt was implanted and the ascites was resolved by the fourth post-operative day. Thirty months later, the vascular end tube of the shunt was ligated. As ascites had not accumulated for 2 weeks, the PV shunt was removed. The patient has been doing well without recurrence of ascites or neuroblastoma for 12 years. As PV shunts were mostly used for long lasting disease, it has not been referred as to how to know when the shunt should be removed. If the shunt is inserted for transient management of ascites, less invasive methods of investigation to know when to remove the shunt need to be developed. PMID- 16477468 TI - [Extensive Fournier gangrene. A dermatologic emergency]. AB - Fournier gangrene is a necrotizing fasciitis of the perineal and genital region, which almost exclusively affects men. The cause is a polymicrobial infection associated with superficial trauma, urological diseases and operations, as well as colorectal diseases. Diabetes mellitus, alcoholism, immunosuppression and other severe illnesses are frequent co-factors. Immediate administration of systemic broad-spectrum antibiotic therapy with coverage of both gram-positive and gram-negative bacteria combined with surgical debridement and intensive medical care can lower the high mortality rate of this condition. PMID- 16477469 TI - [Sebaceous glands as transporters of vitamin E]. AB - Human sebum is produced by sebaceous glands and reaches the skin surface via secretion through the hair shaft. There is experimental evidence that the sebaceous glands and sebum serve as a transport mechanism taking the lipophilic antioxidant vitamin E from the blood to the skin surface. The highest levels of vitamin E are found in the sebum and in the skin lipid film in sebum-rich areas such as facial skin. Recent studies indicate that daily oral supplementation of moderate doses of alpha-tocopherol for at least 3 weeks leads to significant increases of vitamin E levels in human skin sites with a high density of sebaceous glands, such as the face. Thus, the potential photoprotective and antioxidants effects of oral vitamin E, as well as possibly other antioxidants, are site-dependent. These findings should be considered when designing clinical studies to assess the efficacy of oral antioxidants against oxidative stress in the skin. PMID- 16477471 TI - Posttraumatic acute bilateral abducens nerve palsy in a child. AB - INTRODUCTION: Acute bilateral abducens nerve palsy after traumatic clivus epidural haematoma is a rare condition in trauma patients. CASE REPORT: In this report, we present a case with this rare neurological complication of head trauma and discuss the possible mechanisms for abducens nerve palsy and clival epidural haematoma. PMID- 16477470 TI - Hyperforin in St. John's wort drug interactions. AB - Recently, interactions of herbal medicines with synthetic drugs came into focus of particular interest. In the past 3 years, more than 50 papers were published regarding interactions between St. John's wort (Hypericum perforatum L.; SJW) and prescription drugs. Co-medication with SJW resulted in decreased plasma concentrations of a number of drugs including amitriptyline, cyclosporine, digoxin, indinavir, irinotecan, warfarin, phenprocoumon, alprazolam, dextrometorphane, simvastatin, and oral contraceptives. Sufficient evidence from interaction studies and case reports indicate that SJW is a potent inducer of cytochrome P450 enzymes (particularly CYP3A4) and/or P-glycoprotein. Recent studies could show that the degree of enzyme induction by SJW correlates strongly with the amount of hyperforin found in the product. Products that do not contain substantial amounts of hyperforin (<1%) have not been shown to produce clinically relevant enzyme induction. On the other hand, some evidence suggests that hyperforin may also contribute to the antidepressant activity of SJW. However, clinical studies using SJW preparations with a low hyperforin amount (<1%) clearly demonstrated the superiority of this plant extract over placebo and its equivalence to imipramine and fluoxetine in the treatment of mild to moderate forms of depression. In the present paper clinical significant SJW interactions are critically evaluated against the background of hyperforin. PMID- 16477472 TI - Development of a strength test battery for evaluating leg muscle power after anterior cruciate ligament injury and reconstruction. AB - A more sports-specific and detailed strength assessment has been advocated for patients after anterior cruciate ligament (ACL) injury and reconstruction. The purpose of this study was to develop a test battery of lower extremity strength tests with high ability to discriminate between leg power development on the injured and uninjured sides in patients after ACL injury and in patients who have undergone ACL reconstruction. Twenty-three patients were tested 6 months after ACL injury and 44 patients were tested 6 months after ACL reconstruction. Twenty four of the 44 patients were operated on using a hamstrings graft and 20 patients were operated on using a patellar tendon graft. All the patients performed a test battery of three strength tests for each leg in a randomised order. The three strength tests were chosen to reflect quadriceps and hamstring muscular power in a knee-extension and a knee-flexion test (open kinetic chain) and lower-extremity muscular power in a leg-press test (closed kinetic chain). There was a higher sensitivity for the test battery to discriminate abnormal leg power compared with any of the three strength tests individually. Nine out of ten patients after ACL reconstruction and six out of ten of the patients after ACL injury exhibited abnormal leg power symmetry using the test battery. Thus, this test battery had high ability in terms of discriminating between the leg power performance on the injured and uninjured side, both in patients with an ACL injury and in patients who have undergone ACL reconstruction. It is concluded that a test battery consisting of a knee-extension, knee-flexion and leg-press muscle power test had high ability to determine deficits in leg power 6 months after ACL injury and reconstruction. Only a minority of the patients had restored leg muscle power. The clinical relevance is that the test battery may contribute to the decision making process when deciding whether and when patients can safely return to strenuous physical activities after an ACL injury or reconstruction. PMID- 16477473 TI - Life at the edge: an experimental study of a poleward range boundary. AB - Experimental studies of biogeographic processes are important, but rarely attempted because of the logistical challenges of research at large spatial scales. I used a series of large-scale transplant experiments to investigate the mechanisms controlling species abundance near a poleward range boundary. The intertidal limpet Collisella scabra experiences a 100-fold decline in abundance over the northernmost 300 km of its range. Temperature and food supply both strongly influenced individual survival, growth, and maturation. Regression analysis also revealed significant interactions among these conditions: the effect of one could not be predicted without knowing the level of the other. But these relationships could not explain geographic abundance patterns. Instead, individual limpets were highly successful at sites with relatively low abundance. These results suggest that, even though temperature is important to the success of individual C. scabra populations, the primary effect of warming temperatures under climate change may not be a shift in geographic distribution. PMID- 16477477 TI - The population prevalence of child psychiatric disorders in Danish 8- to 9-year old children. AB - The overall prevalence rates of general and specific child psychiatric disorders in Danish children are unknown. In this study, which aimed to estimate prevalence rates, a multi-method strategy using a two-step design was employed. The first step involved assessment with the Child Behaviour Checklist (CBCL). The second step consisted of assessment using the Schedule for Affective Disorders and Schizophrenia for School-Aged Children; Present and Lifetime version (K-SADS-PL), The Children's Global Assessment Scale (C-GAS), The Wechsler Intelligence Scale for Children (WISCIII), The Autism Spectrum Disorder Screening Questionnaire (ASSQ), and a checklist containing the diagnostic criteria for Pervasive Developmental Disorders (PDD). Non-respondents were assessed through teachers using a modified brief version of the K-SADS-PL. A total of 751 children were targeted. The overall estimated prevalence rate of child psychopathology was 11.8 % [95% confidence interval (CI): 8.8, 14.8]. Attention Deficit/ Hyperactivity Disorder (ADHD) was found to be the most common specific child psychiatric disorder. There was no difference in prevalence rates between respondents and non respondents. The estimated prevalence rates were broadly comparable to prevalence rates found in other epidemiological studies. The teacher-based interview proved to be a valid instrument for the assessment of non-respondents. PMID- 16477478 TI - A tribute to Wilhelm H. Erb. PMID- 16477476 TI - Functional properties of the titin/connectin-associated proteins, the muscle specific RING finger proteins (MURFs), in striated muscle. AB - The efficient functioning of striated muscle is dependent upon the proper alignment and coordinated activities of several cytoskeletal networks including myofibrils, microtubules, and intermediate filaments. However, the exact molecular mechanisms dictating their cooperation and contributions during muscle differentiation and maintenance remain unknown. Recently, the muscle specific RING finger (MURF) family members have established themselves as excellent candidates for linking myofibril components (including the giant, multi functional protein, titin/connectin), with microtubules, intermediate filaments, and nuclear factors. MURF-1, the only family member expressed throughout development, has been implicated in several studies as an ubiquitin ligase that is upregulated in response to multiple stimuli during muscle atrophy. Cell culture studies suggest that MURF-1 specifically has a role in maintaining titin M-line integrity and yeast two-hybrid studies point toward its participation in muscle stress response pathways and gene expression. MURF-2 is developmentally down-regulated and is assembled at the M-line region of the sarcomere and with microtubules. Functionally, its expression is critical for maintenance of the sarcomeric M-line region, specific populations of stable microtubules, desmin and vimentin intermediate filaments, as well as for myoblast fusion and differentiation. A recent study also links MURF-2 to a titin kinase-based protein complex that is reportedly activated upon mechanical signaling. Finally, MURF-3 is developmentally upregulated, associates with microtubules, the sarcomeric M line (this report) and Z-line, and is required for microtubule stability and myogenesis. Here, we focus on the biochemical and functional properties of this intriguing family of muscle proteins, and discuss how they may tie together titin mediated myofibril signaling pathways (perhaps involving the titin kinase domain), biomechanical signaling, the muscle stress response, and gene expression. PMID- 16477479 TI - Neuropsychology of multiple sclerosis--an overview. AB - Neuropsychological dysfunctions have been well documented in patients with multiple sclerosis. We herein give an overview of the main findings of recent decades. The pros and cons of considering the cognitive deficits of multiple sclerosis patients as a form of 'subcortical dementia' are discussed. By comparing the neuropsychological descriptions of multiple sclerosis patients as given in the literature to other neurological diseases which are also associated with neuropsychological deficiencies, we propose the term 'multiple disconnection syndrome' to describe the variety of neuropsychological deficits encountered in this demyelinating disease. Furthermore, a ' threshold of cerebral tolerance' is assumed which must be crossed before cognitive dysfunctions are manifested clinically. PMID- 16477481 TI - Multiple sclerosis and the autonomic nervous system. AB - Symptoms related to alterations of the autonomic nervous system are frequent in patients with multiple sclerosis (MS). Bladder or bowel dysfunction or impairment of sexual performance is highly distressing for most MS patients,whereas the clinical relevance of other autonomic symptoms is less clear. Cardiovascular autonomic alterations might relate to clinical signs such as orthostatic intolerance; a relationship with fatigue is uncertain. However, the frequency of abnormal findings in tests for the cardiovascular autonomic system varies due to the lack of standardized test performance or differentially used cut-off values. The incidence of additional symptoms such as pupillomotor or sweating alterations and especially their relationship to overall autonomic nervous system abnormalities is not well known. Although their impact on daily life is low, they can at least serve as diagnostic tools. Beside these clinical aspects, alterations of the autonomic nervous system have also been reported to alter immunological cascades in experimental conditions. However, corresponding results have not been confirmed in clinical trials yet. PMID- 16477480 TI - Botulinum toxin in multiple sclerosis. AB - Local administration of botulinum toxin (BoTx) inhibits presynaptic acetylcholine release. All cholinergically innervated muscles and glands can be paralyzed accordingly. Studies on the application of BoTx in multiple sclerosis have shown good results for focal spasticity, in particular, of the extremities. The first promising data were published on bladder disorders, especially with regard to detrusor sphincter dyssynergia and detrusor hyperreflexia. Hyperhidrosis, hypersalivation and proctologic symptoms might be other areas of application. Approval, however, has been greatly restricted which limits the therapeutic range of use. PMID- 16477482 TI - Botulinum toxin in patients with multiple sclerosis. AB - Nearly all patients with multiple sclerosis (MS) will develop spasticity in the course of their disease. This symptom accounts for most of the handicap and impairment in the quality of life. Treatment with botulinum toxin will enable an efficient and safe alleviation of spasticity and the problems involved, given a realistic definition of the therapeutic target and a graded multimodal approach. Treatment may fail for a great number of reasons that require diligent analysis. Compared with other disorders resulting in spasticity as well, MS does not constitute a monophasic disorder but is influenced by many factors. Treatment of spasticity in MS must therefore be guided by its particular aspects. PMID- 16477483 TI - Identification and management of cognitive disorders in multiple sclerosis--a consensus approach. PMID- 16477485 TI - Current issues in immunomodulatory treatment of multiple sclerosis--a practical approach. PMID- 16477484 TI - Autonomic dysfunction in multiple sclerosis. AB - Multiple sclerosis (MS) is the most frequent chronic neurological disease affecting young persons in developed countries. MS is, however, considered as a secondary cause, of central origin, for autonomic dysfunction. The most common autonomic symptoms in MS are disorders of micturation, impotence, sudomotor and gastrointestinal disturbances, orthostatic intolerance as well as sleep disorders. The majority of the patients suffer at some period of the disease from lower urinary tract symptoms and sexual dysfunction. Awareness and treatment of these conditions is vital to improving health and quality of life in patients with MS. The increased understanding of the pathophysiological mechanisms in autonomic dysfunction in MS, along with technological and pharmaceutical developments has advanced our ability to treat the multiple aspects complicating autonomic failure in MS. PMID- 16477489 TI - Cerebrogenic cardiac arrhythmias: cortical lateralization and clinical significance. AB - That the brain may be involved in cardiovascular regulation has been acknowledged for over a century. That cardiac arrhythmias may result from cortical derangement has been less well recognized. That cortical cardiac representation may be lateralized is even more controversial. Recent evidence implicates several cortical structures, especially the insula, in cardiac rate and rhythm control. Experimental models indicate that insular lesions may be arrhythmogenic. Accumulating data show similar lesion effects in humans. In the rat, monkey and man sympathetic cardiovascular control is generally represented in the right insula, although pronounced insulo-insular connectivity has been demonstrated. Proarrhythmic shifts in cardiac sympathovagal balance occur after human stroke, including left insular lesions. This evidence implicates the cortex in the promotion and even generation of cardiovascular dysfunction under appropriate circumstances. PMID- 16477490 TI - Cardiovascular manifestations of autonomic epilepsy. AB - Cardiovascular autonomic manifestations of seizures occur frequently in the epileptic population. Common manifestations include alterations in heart rate and rhythm, blood pressure, ECG changes and chest pain. The neuroanatomical and neurophysiological underpinnings of these autonomic manifestations are not been fully elucidated. Diagnostic confusion may arise when ictal symptoms are confined to the autonomic nervous system; conversely, such symptoms in association with convulsions or altered consciousness are more readily recognized as concomitant ictal features. Awareness of the diverse autonomic manifestations of epilepsy will enhance diagnosis and lead to more effective therapy of these patients. PMID- 16477491 TI - Seizures and syncope: anatomic basis and diagnostic considerations. AB - Although pathophysiologically distinct, syncope and seizures share clinical characteristics which may make diagnosis difficult. Adding to diagnostic complexity are the facts that seizures and syncope may coexist in the same patient, syncope may be associated with seizure-like motor manifestations, and seizures may be complicated by cardiac arrhythmia and syncope. Combined EEG/ECG telemetry is sometimes necessary to establish the correct diagnosis. These techniques also provide an opportunity to study the role of certain cortical regions in the modulation of cardiac function. There is an increasing understanding of the central autonomic pathways involved in the genesis of the cardiovascular changes that occur during epileptic seizures. This article reviews the use of EEG/ECG telemetry in the evaluation of syncope and seizures, and the neuroanatomic circuitry involved in the production of the cardiovascular manifestations of seizures. PMID- 16477492 TI - Therapeutic effects of vagus nerve stimulation in epilepsy and implications for sudden unexpected death in epilepsy. AB - Vagus nerve stimulation (VNS) is a non-pharmacological therapy approved by the FDA for treatment of patients with partial-onset epilepsy. The most frequently encountered adverse effects typically occur during stimulation, are usually mild to moderate in severity, and resolve with reduction in current intensity or spontaneously over time. There are no apparent effects of VNS on vagally mediated visceral function. Though the precise mechanism of action of VNS remains unknown, available evidence suggests that central autonomic nervous system pathways are involved, which have also been implicated in sudden unexpected death in epilepsy (SUDEP). Studies to date of VNS and SUDEP are limited and do not conclusively show an association between VNS and SUDEP rates independent of other epilepsy specific variables. PMID- 16477493 TI - Subtle involvement of the parasympathetic nervous system in patients with irritable bowel syndrome. AB - This study comprises assessment of autonomic function in irritable bowel syndrome (IBS) patients, focusing on meal-related changes. In 18 IBS patients (4 males, mean age 45+/-3.0 [SEM] years) and 19 healthy volunteers (6 males, mean age 41+/ 3.5 years) blood pressure, heart rate, heart rate variability and muscle sympathetic nerve activity (MSNA) were assessed before, during and after consumption of a standardized meal. In pre- and postprandial phase Valsalva maneuver, cold pressor test (CPT) and deep breathing test were carried out and Visual Analog Scale (VAS) scores for nausea, bloating and pain were obtained. In the IBS group, the meal induced significantly higher VAS scores for pain (P=0.002) and bloating (P=0.02). During food intake, the increase in blood pressure, heart rate and MSNA was equal in patients and controls, but the increase of LF/HF ratio of heart rate variability was significantly higher in the IBS group (median [quartiles] 2.29 [1.14-3.00] versus 0.77 [0.25-1.81]; P=0.03). IBS patients scored lower on pre- and postprandial RRmax/RRmin ratio during deep breathing (DB ratio, P=0.03). The increase in MSNA (burst frequency) in response to CPT tended to be higher in the IBS patients (P=0.07). We conclude that reactivity to food intake, measured as muscle sympathetic nerve activity, is normal in IBS patients. The lower DB ratio and higher LF/HF ratio during food intake in IBS patients is an indication of a reduced parasympathetic reactivity. These results suggest that reduced baseline activity as well as responsiveness of the parasympathetic system could play a role in the pathogenesis of IBS. PMID- 16477494 TI - Plasma catecholamines and blood volume in native Andeans during hypoxia and normoxia. AB - Plasma catechols and blood volume were measured in 20 male, native high-altitude residents of Cerro de Pasco, Peru (4338 m), while hypoxic and subsequently while normoxic at sea level. Ten subjects were healthy controls,with hematocrits lower than 61%, and ten had chronic mountain sickness (CMS), a syndrome of maladaptation to altitude, characterized by polycythemia (hematocrit > 61%), profound hypoxemia, and neurologic symptoms. The main aim of the study was to evaluate the chronic effects of hypoxia on plasma catechols and on blood volume, by studying these parameters during hypoxia at high altitude (HA) and shortly after exposure to normoxia at sea level (SL). Subjects were first studied at HA in their habitual hypoxic environment, and measurements were repeated within 4 hours of arrival at SL (Lima, Peru, 150 m). All subjects had higher plasma norepinephrine (NE), dopamine (DA), and dihydroxyphenylglycol (DHPG) levels in HA (NE in controls and CMS: 414+/-47 and 514+/-35 pg/mL; DA: 9+/-1 and 13+/-1 pg/mL, DHPG: 817+/-48 and 972+/-77 pg/mL) than at SL (NE: 164+/-9 and 243+/-28 pg/mL; DA: 4+/-0.5 and 5+/-1 pg/mL DHPG: 502+/-23 and 649+/-39 pg/mL). Group differences were statistically significant only for NE in the CMS group. Plasma volume was higher in HA in both groups (p<0.05); red cell volume was higher in HA only in the CMS group. The results indicate sympathetic nervous stimulation by chronic ambient hypoxia at altitude in Andean natives, independent of maladaptation to their native environment. PMID- 16477496 TI - Acute effects of C-peptide on gastric emptying in longstanding type 1 diabetes. AB - Gastric emptying (GE) of a solid (100 g beef) and liquid (150 ml 10 % dextrose) meal was measured in eight patients with type 1 diabetes during intravenous infusion of C-peptide (6 pmol/kg/ min) or isotonic saline. C-peptide had no effect on either solid or liquid GE. PMID- 16477497 TI - Left-right asymmetry of the facial microvascular control. AB - Facial blood flow and temperature were significantly higher on the right side of the forehead compared to the left. This asymmetry implies that the hemispheric autonomic control of the face differs and could influence the expression of emotion. PMID- 16477495 TI - Orthostatic hypotension as an early finding in Parkinson's disease. AB - Patients with Parkinson's disease (PD) commonly have clinically significant orthostatic hypotension (OH). In such patients PD+OH might be confused with multiple system atrophy (MSA), in which OH is a frequent finding, or with pure autonomic failure (PAF), if OH preceded clinical manifestations of the movement disorder. This study addressed whether OH can occur as an early finding in PD+OH. Historical data were analyzed from 35 patients with PD+OH evaluated at the NIH. OH was considered early if the patient had OH before, concurrent with, or starting within 1 year after onset of a symptomatic movement disorder. MSA was excluded by myocardial 6-[(18)F]fluorodopamine-derived radioactivity more than 2 standard deviations below the normal mean. Among the 35 PD+OH patients, 21 (60 %) had documentation of OH as an early finding. In 4 such patients, OH had preceded parkinsonism, and in 4 others, OH had dominated the early clinical picture, even after cessation of levodopa treatment for the movement disorder. In PD, OH can occur early in the disease, occasionally preceding or overshadowing the movement disorder. PMID- 16477498 TI - The effects of exercise training on myocardial adrenergic and muscarinic receptors. AB - We investigated the effects of exercise training on heart rate variability (HRV) and myocardial adrenergic and muscarinic receptors in rats. Exercise training induced a decrease in body mass while ventricular size remained unchanged, a development we considered as a relative cardiac hypertrophy. In addition, there was a reduction in the density of myocardial beta(1)-adrenergic receptors. These structural changes were associated with functional adaptations, as illustrated by the increased response of the sinus node to sympathetic blockade. PMID- 16477500 TI - Ischemic electrocardiographic change induced by exercise in a patient with chronic autonomic failure. AB - We report a patient with chronic autonomic failure, who had exercise-induced ischemic electrocardiographic changes without coronary artery abnormalities. Orthostatic hypotension and exercise-induced release of humoral vasoconstrictors may have caused the myocardial ischemia. PMID- 16477499 TI - Urinary dysfunction and autonomic control in amyloid neuropathy. AB - Uro-neurological assessment was performed in four patients with small-fiber neuropathy due to amyloidosis (2 transthyretin-type/2 immunoglobulin light-chain type). Voiding difficulties were due to detrusor weakness and impaired bladder sensation. In two patients cholinesterase inhibition treatment caused urge incontinence, indicating detrusor denervation supersensitivity. The underlying mechanisms of urinary dysfunction seem to involve postganglionic cholinergic and afferent somatic nerves. PMID- 16477501 TI - Fainting in your sleep? PMID- 16477502 TI - Hemodynamic computation using multiphase flow dynamics in a right coronary artery. AB - Hemodynamic data on the roles of physiologically critical blood particulates are needed to better understand cardiovascular diseases. The blood flow patterns and particulate buildup were numerically simulated using the multiphase non-Newtonian theory of dense suspension hemodynamics in a realistic right coronary artery (RCA) having various cross sections. The local hemodynamic factors, such as wall shear stress (WSS), red blood cell (RBC) buildup, viscosity, and velocity, varied with the spatially nonuniform vessel structures and temporal cardiac cycles. The model generally predicted higher RBC buildup on the inside radius of curvature. A low WSS region was found in the high RBC buildup region, in particular, on the area of maximum curvature of a realistic human RCA. The complex recirculation patterns, the oscillatory flow with flow reversal, and vessel geometry resulted in RBC buildup due to the prolonged particulate residence time, specifically, at the end of the diastole cycle. The increase of the initial plasma viscosity caused the lower WSS. These predictions have significant implications for understanding the local hemodynamic phenomena that may contribute to the earliest stage of atherosclerosis, as clinically observed on the inside curvatures and torsion of coronary arteries. PMID- 16477503 TI - Characterization of superparamagnetic nanoparticle interactions with extracellular matrix in an in vitro system. AB - Controlled dispersion of therapeutic agents within liquid- and gel-filled cavities represents a barrier to treatment of some cancers and other pathological states. Interstitial delivery is compromised by the poor mobility of macromolecules and larger nanoscale structures. We developed an in vitro system to quantify the suitability of superparamagnetic nanoparticles (SPM NPs) as a site-specific therapeutic vehicle for delivery through fluid- and gel-based systems. SPM NP motion was induced by an external magnetic field. NP migration was modulated by NP concentration and surface coating. 135 nanometer radius PEGylated NPs moved through the extracellular matrix with an average velocity of 1.5 mm h(-1), suitable for some clinical applications. Increasing the SPM NP radius to 400 nm while maintaining the same per NP magnetic susceptibility resulted in a greater than 1,000-fold reduction in magnetic mobility, to less than 0.01 mm h(-1). The critical influence of NP size on gel permeation was also observed in silica-coated 135 nm SPM NPs that aggregated under the experimental conditions. Aggregation played a critical role in determining the behavior of the nanoparticles. SPM NPs allow significant free-solution mobility to specific sites within a cavity and generate sufficient force to penetrate common in vivo gels. PMID- 16477506 TI - Absolute measurements of photoluminescence quantum yields of solutions using an integrating sphere. AB - We demonstrate that absolute measurements of the photoluminescence quantum yield of solutions can be made using an integrating sphere and a conventional fluorimeter. With this method the need for measurements against a luminescence standard is overcome. The sphere is mounted inside a commercial fluorimeter, which gives flexibility in excitation and emission wavelength ranges. A number of compounds have been investigated and the results are compared to literature values and data obtained using a comparative method. PMID- 16477505 TI - Investigation of novel quantum dots/proteins/cellulose bioconjugate using NSOM and fluorescence. AB - We investigated the engineered bioconjugate of cadmium selenide core/zinc sulfide shell, (CdSe)ZnS, quantum dots (QDs) with genetically modified proteins using fluorescence spectroscopy, near-field scanning optical microscopy (NSOM) and spectroscopy (NSOS). The protein polymer was allowed to self-assemble to the bacterial microcrystalline cellulose surface through the cellulosic binding domain. Results from the sample containing the QDs/protein/cellulose assemblies suggest that QDs were arrayed along the cellulose surface. The spectroscopic change of spectroscopic properties of the QDs upon bioconjugation, indicating the interaction among the immobilized QDs and between the constructed protein and QDs. PMID- 16477507 TI - Fibrinogen-beta-estradiol binding studied by fluorescence spectroscopy: denaturation and pH effects. AB - Fibrinogen is a blood plasma protein that plays a crucial role in hemostasis. It is known that erythrocyte aggregation increases in the presence of fibrinogen, and that beta-estradiol decreases erythrocyte aggregation with a constant fibrinogen concentration. In this work, we have used intrinsic tryptophan fluorescence to obtain information on the conformational changes of fibrinogen upon the recently proposed interaction with beta-estradiol. To evaluate the effect on the conformational changes during fibrinogen-beta-estradiol binding, fluorescence experiments were performed using guanidine hydrochloride (0-6 M) as denaturant, at different pH values. The results obtained for pH 6.5 and 8.0 showed no effect during the binding. The main differences were observed between pH 4.2 and 7.4, in the absence and in the presence of two different denaturant concentrations (1 and 5 M). A red shift of the fluorescence emission from 344 to 354 nm is observed when denaturant concentration is above 3 M for all studied pH values. This phenomenon may be explained by the loss of compact structure of the protein in the presence of denaturant, with tryptophan residues exposure to the aqueous environment and alteration of fibrinogen-beta-estradiol binding. These results demonstrate that the binding sites of fibrinogen are strongly dependent on the conformational state of the protein. PMID- 16477508 TI - Relation between fluorescence decays and temporal evolution of excited states. AB - A differential equation system describing the temporal evolution of excited substates and fluorescence emission were tested using a DOPRI algorithm. The numerical solutions show that there is significant difference in the measurable parameters according to the type of connectivity among the excited substates. In the globally connected case, the fluorescence emission exhibits a double exponential behavior, and the first moment of the emitted spectrum decays with stretched exponential characterized by beta < 1. In the diffusive case the fluorescence emission cannot be always fitted with double exponential, and the first moment of the emitted spectrum may decay with stretched exponential characterized by beta > 1. Details of modeling and the possibilities of drawing conclusions are also presented. PMID- 16477510 TI - Condom use and its correlates among female sex workers in Hanoi, Vietnam. AB - Knowledge of female sex workers' (FSW) condom use behaviors in Vietnam is important for predicting the epidemic and designing interventions. Four hundred FSWs in Hanoi were studied in 2002. Consistent condom use in the past month was higher with irregular clients (62%), less with regular clients (41%), and lowest with "love mates" (5%). Reasons for not using condoms were partner objection, condom unavailability, and belief of partner's disease-free status. Twenty-seven percent reported not always having a condom available. Thirty-five percent reported increasing condom use in the previous 6 months. Reluctance to ask clients to use condoms and condom unavailability were independently associated with inconsistent condom use with both irregular and regular clients. Older age was also associated with inconsistent condom use with irregular clients. Condom promotion should focus on FSWs and their partners. Negative attitudes toward FSWs and condom promotion need to be changed to reduce stigmatization of FSWs and to make condom use a norm in the society. PMID- 16477511 TI - Additive deleterious effects of methamphetamine dependence and immunosuppression on neuropsychological functioning in HIV infection. AB - Methamphetamine (MA) dependence and HIV infection are independently associated with cerebral dysfunction, especially within frontal-basal ganglia circuits. Recent evidence indicates that MA dependence has an additive effect on neuropsychological (NP) deficits associated with HIV infection. This study extends prior findings by examining the combined effects of MA dependence (MA+) and immunosuppression (i.e., CD4 lymphocyte count <200) on NP functioning in 284 HIV+ individuals. Prevalence of NP impairment was examined in four demographically comparable groups: (1) MA+/CD4 < 200; (2) MA+/CD4 > or = 200; (3) MA-/CD4 < 200; and (4) MA-/CD4 > or = 200. Results revealed that both MA dependence and immunosuppression were significant predictors of NP impairment. More importantly, additive effects were evident whereby the MA+/CD4 < 200 group exhibited the highest rate of NP impairment. Findings indicate that MA dependence conveys an additive deleterious impact on NP status in immunosuppressed persons with HIV infection, perhaps reflecting the combined effects of neuropathophysiological mechanisms in fronto-striatal circuits. PMID- 16477509 TI - Use of fluorescence probes for detection of reactive nitrogen species: a review. AB - The biological and toxicological effects that have been attributed to reactive nitrogen species (RNS) are increasingly stirring the scientific inquisitiveness about the molecular mechanisms involved. However, RNS present some characteristics that complicate their detection, namely their short lifetime and the normal presence of a variety of endogenous compounds capable of reacting with these reactive species, when the studies are performed in biological matrices. The development of methodologies capable of circumvent these difficulties is thus of fundamental importance. Fluorescence probes are particularly important due to their high sensibility and usefulness in temporal and spatial monitoring of RNS, particularly in microanalysis conditions in biological media akin to cells or tissues. In the present review is given an account of the fluorescence probes that have been used for detection of nitric oxide (*NO), peroxynitrite anion (ONOO-), as well as of some of its derivatives in biological and nonbiological media. PMID- 16477512 TI - Experts outline ways to decrease the decade-long yearly rate of 40,000 new HIV infections in the US. AB - This paper presents data from a brief, anonymous, open-ended survey of 50 behavioral research experts in HIV prevention. Responses were received from 31 participants who provided input regarding the primary reasons they believe the rate of the HIV epidemic in the United States has persisted in recent years, and how they believe we can most efficiently decrease the current rate of new HIV infections in the United States. Four clusters of reasons suggested for the persistent rate of new infections: Intervention level reasons, Society level reasons, Person level reasons, and Multiple Risk Factor reasons. Three clusters of strategies suggested for decreasing the current rate: Improved Targeting of HIV Prevention efforts, Larg-Scale Changes to HIV prevention, and Integrating HIV Prevention into more aspects of society. Results are reviewed with the objective of providing a fresh perspective on the potential means for addressing the current HIV epidemic. PMID- 16477514 TI - Effects of low-intensity behavioral treatment for children with autism and mental retardation. AB - We retrospectively compared 2 groups of children receiving either behavioral treatment (n = 13) or eclectic treatment (n = 15) for an average of 12 hours per week. Children were assessed on intelligence, language, adaptive functioning and maladaptive behavior at pretreatment and 2 years into treatment. The groups did not differ significantly at pretreatment. After 2 years of treatment, the behavioral group made larger gains than the eclectic group in most areas. However, gains were more modest than those reported in previous studies with children receiving more intensive behavioral treatment, and it is questionable whether they were clinically significant. PMID- 16477513 TI - ADHD symptom subtypes in children with pervasive developmental disorder. AB - OBJECTIVE: Compares DSM-IV ADHD subtypes in large samples of 3-to-5 and 6-to-12 year old children with pervasive developmental disorder (PDD) vs. nonPDD clinic referrals. METHOD: Parents and teachers completed a DSM-IV-referenced rating scale. RESULTS: ADHD subtypes were clearly differentiated from the nonADHD group and showed a differential pattern of co-occurring psychiatric symptoms (more pronounced for teacher- than parent-defined subtypes and older than younger children) that was similar in both PDD and nonPDD samples. The Combined type had more severe oppositional, aggressive, and PDD symptoms than Inattentive type and were from less advantaged homes than other subtypes. Hyperactive-Impulsive type was least impaired. CONCLUSION: Findings support the notion that ADHD may be a clinically meaningful syndrome in children with PDD. PMID- 16477515 TI - The Cambridge Mindreading (CAM) Face-Voice Battery: Testing complex emotion recognition in adults with and without Asperger syndrome. AB - Adults with Asperger Syndrome (AS) can recognise simple emotions and pass basic theory of mind tasks, but have difficulties recognising more complex emotions and mental states. This study describes a new battery of tasks, testing recognition of 20 complex emotions and mental states from faces and voices. The battery was given to males and females with AS and matched controls. Results showed the AS group performed worse than controls overall, on emotion recognition from faces and voices and on 12/20 specific emotions. Females recognised faces better than males regardless of diagnosis, and males with AS had more difficulties recognising emotions from faces than from voices. The implications of these results are discussed in relation to social functioning in AS. PMID- 16477516 TI - Motion and form coherence detection in autistic spectrum disorder: Relationship to motor control and 2:4 digit ratio. AB - Children with autistic spectrum disorder and controls performed tasks of coherent motion and form detection, and motor control. Additionally, the ratio of the 2nd and 4th digits of these children, which is thought to be an indicator of foetal testosterone, was measured. Children in the experimental group were impaired at tasks of motor control, and had lower 2D:4D than controls. There were no group differences in motion or form detection. However a sub-group of children with autism were selectively impaired at motion detection. There were significant relationships between motion coherence detection and motor control in both groups of children, and also between motion detection, fine motor control and 2D:4D in the group of children with autistic spectrum disorder. PMID- 16477518 TI - Management of mental disorders in VA primary care practices. AB - The association between facility-level organizational features and management of mental health services was assessed based on a survey of directors from 219 VA primary care facilities. Overall, 26.4% of VA primary care facilities referred patients with depression, while 72.6% and 46.1% referred patients with serious mental illness and substance use disorders, respectively Staffing mix (i.e., physician extenders such as nurse practitioners) was associated with a lesser likelihood of mental health referral. Managed care (preauthorization requirement) was associated with a greater likelihood of referral for depression. VA primary care programs, while tending to refer for more serious mental illnesses, may also be using mental health specialists and physician extenders to provide mental health care within general medical settings. PMID- 16477519 TI - The implications of simultaneous smoking initiation for inferences about the genetics of smoking behavior from twin data. AB - We examined early social influences across stages of smoking within the context of a twin study using an environmental exposure specific to smoking: whether twins started smoking at the same time ("simultaneous smoking initiation": SSI). We expected that SSI would be a good index of shared social influences on smoking initiation. Rates of SSI were indeed significantly higher in MZ twins and in twins who shared peers and classes, as well as in male twins. With the exception of regular smoking in females, we found no significant difference in estimates of genetic and environmental parameters between SSI and non-SSI pairs for any of the smoking measures that we examined (DSM-IV and Fagerstrom HSI measures of nicotine dependence; DSM-IV nicotine withdrawal; heavy smoking; and in males, regular smoking). For regular smoking in females, allowing for additional shared environmental influences associated with SSI only modestly reduced our estimates of additive genetic variance (56% vs. 68%). These results indicate the important social influences that may occur for smoking initiation do not appear to seriously bias estimates of genetic effects on later stages of smoking. PMID- 16477520 TI - A further analysis of sexual isolation between sympatric and allopatric populations of Drosophila Pseudoobscura and D. persimilis : rejoinder to Noor and Ortiz-Barrientos. PMID- 16477517 TI - Impaired face processing in autism: fact or artifact? AB - Within the last 10 years, there has been an upsurge of interest in face processing abilities in autism which has generated a proliferation of new empirical demonstrations employing a variety of measuring techniques. Observably atypical social behaviors early in the development of children with autism have led to the contention that autism is a condition where the processing of social information, particularly faces, is impaired. While several empirical sources of evidence lend support to this hypothesis, others suggest that there are conditions under which autistic individuals do not differ from typically developing persons. The present paper reviews this bulk of empirical evidence, and concludes that the versatility and abilities of face processing in persons with autism have been underestimated. PMID- 16477523 TI - Conspecific and interspecific interactions between the FEM-2 and the FEM-3 sex determining proteins despite rapid sequence divergence. AB - Using degenerate oligonucleotide primers, we isolated the Caenorhabditis remanei orthologue of the C. elegans sex-determining phosphatase gene fem-2 as well as two other protein phosphatase homologues. Despite the significant sequence divergence between C. elegans and C. remanei FEM-2, we used RNAi-mediated gene knockdown to demonstrate that at least some aspects of male development require FEM-2 function in C. remanei. Consistent with this functional conservation, the conspecific interaction between the FEM-2 and the FEM-3 proteins observed in C. elegans also occurs in C. remanei. To further explore whether the rapid evolution of FEM-2 and FEM-3 affects their molecular interactions, we tested for cross species interactions between the proteins from C. elegans, C. briggsae, and C. remanei. Although all FEM-2/FEM-3 pairs from a single species interact, only two out of six interspecific pairs bind each other, showing that FEM-2 and FEM-3 are coevolving. Both interspecific interactions involved C. briggsae FEM-3. We constructed chimeric versions of FEM-2 consisting of various combinations of the C. elegans and C. remanei proteins. C. briggsae FEM-3 interacted with all the chimeras, even those that did not interact with either C. elegans or C. remanei FEM-3. We hypothesize that the promiscuity of C. briggsae FEM-3 reflects an increased reliance on evolutionarily constrained regions of FEM-2 for binding. If so, our data support the notion that the coevolution of two interacting proteins sometimes involves a shift in the domains that contribute to binding. PMID- 16477524 TI - The response of amino acid frequencies to directional mutation pressure in mitochondrial genome sequences is related to the physical properties of the amino acids and to the structure of the genetic code. AB - The frequencies of A, C, G, and T in mitochondrial DNA vary among species due to unequal rates of mutation between the bases. The frequencies of bases at fourfold degenerate sites respond directly to mutation pressure. At first and second positions, selection reduces the degree of frequency variation. Using a simple evolutionary model, we show that first position sites are less constrained by selection than second position sites and, therefore, that the frequencies of bases at first position are more responsive to mutation pressure than those at second position. We define a measure of distance between amino acids that is dependent on eight measured physical properties and a similarity measure that is the inverse of this distance. Columns 1, 2, 3, and 4 of the genetic code correspond to codons with U, C, A, and G in their second position, respectively. The similarity of amino acids in the four columns decreases systematically from column 1 to column 2 to column 3 to column 4. We then show that the responsiveness of first position bases to mutation pressure is dependent on the second position base and follows the same decreasing trend through the four columns. Again, this shows the correlation between physical properties and responsiveness. We determine a proximity measure for each amino acid, which is the average similarity between an amino acid and all others that are accessible via single point mutations in the mitochondrial genetic code structure. We also define a responsiveness for each amino acid, which measures how rapidly an amino acid frequency changes as a result of mutation pressure acting on the base frequencies. We show that there is a strong correlation between responsiveness and proximity, and that both these quantities are also correlated with the mutability of amino acids estimated from the mtREV substitution rate matrix. We also consider the variation of base frequencies between strands and between genes on a strand. These trends are consistent with the patterns expected from analysis of the variation among genomes. PMID- 16477526 TI - Genes expressed in a turrid venom duct: divergence and similarity to conotoxins. AB - The toxoglossate mollusks are a large group of venomous animals (>10,000 species) conventionally divided into three groups, the cone snails, the auger snails, and the turrid snails; turrids account for >90% of the biodiversity of toxoglossans. Only the venoms of cone snails have been intensively investigated, with little work focused on turrids. We report the first broad characterization of genes expressed in venom ducts of any turrid species. Twenty-three different cDNA clones encoding putative toxins were characterized from the venom duct of the turrine species Lophiotoma olangoensis Olivera 2002 and belong to 16 different gene families. Of the 16 different Lophiotoma olangoensis gene families that encode putative toxins, for only 1 was there clear evidence of sequence similarity with any conotoxin gene family. The I-like gene family of Lophiotoma olangoensis was found to be related to the K channel-targeted I(2) conotoxin superfamily. Most putative Lophiotoma toxins are cysteine-rich polypeptides, with a significant fraction much larger (>80 amino acids) than the toxins from cone snails. A small number were not cysteine-rich but had hydrophobic amino acid clusters interspersed with arginine residues. This is only 1 of >10,000 different turrid venoms that needs to be characterized. From this study, a common origin with Conus for one family of putative turrid toxins is indicated. PMID- 16477527 TI - High-definition television in medicine. PMID- 16477525 TI - Molecular evolution of cytochrome c oxidase in high-performance fish (teleostei: Scombroidei). AB - The 13 peptides encoded by vertebrate mitochondrial DNA (mtDNA) are essential subunits of oxidative phosphorylation (OXPHOS) enzymes. These genes normally experience purifying selection and also coevolve with nuclear-encoded subunits of OXPHOS complexes. However, the role of positive selection on mtDNA evolution is still unclear, as most examples of intergenomic coevolution appear to be the result of compensation by nuclear-encoded genes for mildly deleterious mtDNA mutations, and not simultaneous positive selection in both genomes. Organisms that have experienced strong selective pressures to increase aerobic capacity or adapt to changes in thermal environment may be better candidates in which to examine the impact of positively selected changes on mtDNA evolution. The tuna (suborder Scombroidei, family Scombridae) and billfish (suborder Scombroidei, families Xiphiidae and Istiophoridae) are highly aerobic fish with multiple specializations in muscle energetics, including a high mitochondrial content and regional endothermy. We examined the role of positively selected mtDNA substitutions in the production of these unique phenotypes. Focusing on a catalytic subunit of cytochrome c oxidase (COX II), we found that the rate ratio of nonsynonymous (d(N); amino acid changing)-to-synonymous (d(S); silent) substitutions was not increased in lineages leading to the tuna but was significantly increased in the lineage preceding the billfish. Furthermore, there are a number of individual positively selected sites that, when mapped onto the COX crystal structure, appear to interact with other COX subunits and may affect OXPHOS function and regulation in billfish. PMID- 16477529 TI - Intracellular Mg2+ influences both open and closed times of a native Ca2+ activated BK channel in cultured human renal proximal tubule cells. AB - Effects of intracellular Mg2+ on a native Ca(2+)-and voltage-sensitive large conductance K+ channel in cultured human renal proximal tubule cells were examined with the patch-clamp technique in the inside-out mode. At an intracellular concentration of Ca2+ ([Ca2+](i)) of 10(-5)-10(-4) M, addition of 1 10 mM: Mg2+ increased the open probability (P(o)) of the channel, which shifted the P(o) -membrane potential (V(m)) relationship to the negative voltage direction without causing an appreciable change in the gating charge (Boltzmann constant). However, the Mg(2+)-induced increase in P(o) was suppressed at a relatively low [Ca2+](i) (10(-5.5)-10(-6) M). Dwell-time histograms have revealed that addition of Mg2+ mainly increased P(o) by extending open times at 10(-5) M Ca2+ and extending both open and closed times simultaneously at 10(-5.5) M Ca2+. Since our data showed that raising the [Ca2+](i) from 10(-5) to 10(-4) M increased P(o) mainly by shortening the closed time, extension of the closed time at 10(-5.5) M Ca(2+) would result from the Mg(2+)-inhibited Ca(2+)-dependent activation. At a constant V(m), adding Mg2+ enhanced the sigmoidicity of the P(o) [Ca2+](i) relationship with an increase in the Hill coefficient. These results suggest that the major action of Mg2+ on this channel is to elevate P(o) by lengthening the open time, while extension of the closed time at a relatively low [Ca2+](i) results from a lowering of the sensitivity to Ca2+ of the channel by Mg2+, which causes the increase in the Hill coefficient. PMID- 16477531 TI - Slow sodium channel inactivation and use-dependent block modulated by the same domain IV S6 residue. AB - Voltage- and/or conformation-dependent association and dissociation of local anesthetic-class drugs from a putative receptor site in domain IV S6 of the sodium channel and slow conformation transitions of the drug-associated channel have been proposed as mechanisms of use- and frequency-dependent reduction in sodium current. To distinguish these possibilities, we have explored the reactivity to covalent modification by thiols and block of the mutations F1760C and F1760A at the putative receptor site of the cardiac sodium channel expressed as stable cell lines in HEK-293 cells. Both mutations decreased steady-state fast inactivation, shifting V1/2h from -86 +/- 1.3 mV (WT) to -72.3 +/- 1.4 mV (F1760C) and -67.7 +/- 1 mV (F1760A). In the absence of drug, the F1760C mutant channel displayed use-dependent current reduction during pulse-train stimulation, and faster onset of slow inactivation. This mutant also retained some sensitivity to lidocaine. In contrast, the F1760A mutant showed no use-dependent current reduction or sensitivity to lidocaine. The covalent-modifying agent MTS-ET enhanced use-dependent current reduction of the F1760C mutant channel only. The use-dependent reduction in current of the covalently modified channel completely recovered with rest. Lidocaine produced no additional block during exposure to MTS-ET-treated cells (MTS-ET 43 +/- 2.7%: MTS-ET lidocaine 47 +/- 4.5%), implying interaction at a common binding site. The data suggest that use-dependent binding at the F1760 site results in enhanced slow inactivation rather than alteration of drug association and dissociation from that site and may be a general mechanism of action of sodium-channel blocking agents. PMID- 16477528 TI - Critical role of tight junctions in drug delivery across epithelial and endothelial cell layers. AB - Epithelia in multicellular organisms constitute the frontier that separates the individual from the environment. Epithelia are sites of exchange as well as barriers, for the transit of ions and molecules from and into the organism. Therapeutic agents, in order to reach their target, frequently need to cross epithelial and endothelial sheets. Two routes are available for such purpose: the transcellular and the paracellular pathways. The former is employed by lipophilic drugs and by molecules selectively transported by channels, pumps and carriers present in the plasma membrane. Hydrophilic molecules cannot cross biological membranes, therefore their transepithelial transport could be significantly enhanced if they moved through the paracellular pathway. Transit through this route is regulated by tight junctions (TJs). The discovery in recent years of the molecular mechanisms of the TJ has allowed the design of different procedures to open the paracellular route in a reversible manner. These strategies could be used to enhance drug delivery across epithelial and endothelial barriers. The procedures employed include the use of peptides homologous to external loops of integral TJ proteins, silencing the expression of TJ proteins with antisense oligonucleotides and siRNAs as well as the use of toxins and proteins derived from microorganisms that target TJ proteins. PMID- 16477530 TI - Contrasting effects of Cd2+ and Co2+ on the blocking/unblocking of human Cav3 channels. AB - Inorganic ions have been used widely to investigate biophysical properties of high voltage-activated calcium channels (HVA: Ca(v)1 and Ca(v)2 families). In contrast, such information regarding low voltage-activated calcium channels (LVA: Ca(v)3 family) is less documented. We have studied the blocking effect of Cd2+, Co2+ and Ni2+ on T-currents expressed by human Ca(v)3 channels: Ca(v)3.1, Ca(v)3.2, and Ca(v)3.3. With the use of the whole-cell configuration of the patch clamp technique, we have recorded Ca2+ (2 mM: ) currents from HEK-293 cells stably expressing recombinant T-type channels. Cd2+ and Co2+ block was 2- to 3 fold more potent for Ca(v)3.2 channels (EC50 = 65 and 122 microM, respectively) than for the other two LVA channel family members. Current-voltage relationships indicate that Co2+ and Ni2+ shift the voltage dependence of Ca(v)3.1 and Ca(v)3.3 channels activation to more positive potentials. Interestingly, block of those two Ca(v)3 channels by Co2+ and Ni2+ was drastically increased at extreme negative voltages; in contrast, block due to Cd2+ was significantly decreased. This unblocking effect was slightly voltage-dependent. Tail-current analysis reveals a differential effect of Cd2+ on Ca(v)3.3 channels, which can not close while the pore is occupied with this metal cation. The results suggest that metal cations affect differentially T-type channel activity by a mechanism involving the ionic radii of inorganic ions and structural characteristics of the channels pore. PMID- 16477532 TI - Expression of glucose-6-phosphatase system genes in murine cortex and hypothalamus. AB - The glucose-6-phosphatase (G6Pase) system participates in the regulation of glucose homeostasis by converting glucose-6-phosphate (G6P) into glucose and inorganic phosphates. We have used an RT-PCR-based cloning and sequencing approach to study the expression of components of the G6Pase system in the hypothalamus and cortex tissues of the ob/ob mouse. We observed the expression of hepatic G6Pase catalytic subunit, G6PC, in both tissues, although increased template inputs were required for its detection. Conversely, expression of both the mouse homologue of the previously-described brain-specific G6P translocase T1 (G6PT1) variant and of the hepatic G6PT1 isoform was easily detectable in hypothalamus and cortex tissues. Of the proposed G6Pase catalytic subunit homologues, the expression of murine ubiquitous G6Pase catalytic subunit-related protein (UGRP, G6PC3) was also easily detectable in both tissues. However, islet specific G6Pase catalytic subunit-related protein (IGRP, G6PC2) was expressed in a tissue-specific manner, and was detectable only in hypothalamus tissue at increased template inputs. We conclude that cells within ob/ob mouse hypothalamus and cortex tissues express genes with either established or proposed roles in G6P hydrolysis. PMID- 16477533 TI - Real-time PCR: housekeeping genes in the INS-1E beta-cell line. AB - Investigation of gene expression is a developing area with several methods available. One method is quantitative PCR. A major pitfall in quantitative PCR is the normalisation procedure of the gene expression. Many experiments include a housekeeping gene, some use RNA concentration, and others use a geometric mean of several internal, stably expressed genes. This study demonstrates that real-time PCR results differ with varying housekeeping genes and analysis protocols when applied to insulin-secreting INS-1E cells derived from the pancreas and stimulated by DEDTC (diethyldithiocarbamate, a zinc chelator) and GLP-1. PMID- 16477534 TI - Nicotinamide increases thyroid radiosensitivity by stimulating nitric oxide synthase expression and the generation of organic peroxides. AB - Differentiated thyroid cancer and hyperthyroidism are treated with radioiodine. However, when the radioisotope dose exceeds certain limits, the patient must be hospitalized to avoid contact with people that would otherwise be exposed to radiation. It would be desirable to obtain a similar therapeutic effect using lower radioiodine doses. Radiosensitizers can be utilized for this purpose. Nicotinamide (NA) increases thyroid radiosensitivity to 131I in both normal and goitrous glands. NA causes a significant increase in thyroid blood flow, which would increase tissue oxygenation and tissue damage via free radicals. Wistar rats were treated with either nicotinamide (NA), 131I or both. The expression of the three isoforms of nitric oxide synthase (NOS) in the thyroid (Western blot) and the activities of SOD, GPx, catalase and organic peroxides were determined. Treatment with NA or 131I increased the expression of eNOS and the generation of organic peroxides. When administered jointly, they showed a synergistic effect. No changes were observed in the other NOS isoforms or in the activities of catalase, glutathione peroxidase and superoxide dismutase. NA potentiates the effect of 131I by increasing eNOS, which would in turn stimulate NO production, increasing thyroid blood flow and tissue damage via organic peroxides. PMID- 16477535 TI - Effect of short-term therapy with recombinant human growth hormone (GH) on metabolic parameters and preclinical atherosclerotic markers in hypopituitary patients with growth hormone deficiency. AB - OBJECTIVE: This study examines the effects of growth hormone replacement on body composition, insulin sensitivity, lipid profile, endothelial dysfunction and carotid intima media thickness in patients with adult-onset growth-hormone (GH) deficiency. METHODS: Twelve patients with severe GH deficiency received GH replacement for one year. In all patients, the following parameters were evaluated before and after six and twelve months of therapy: fasting glucose, insulin levels and lipid profile, bone mineral density and body composition. Carotid intima media thickness and brachial flow-mediated dilatation were also evaluated by arterial ultrasonography at basal condition and after one year of therapy. RESULTS: No significant changes were seen in body weight and blood pressure, total fat and lean mass, or bone mineral density after six months of GH replacement. There was an increase in triglycerides (p = 0.05), while total and HDL cholesterol, blood glucose, insulin levels did not change significantly. After twelve months, an increase in lean mass and a decrease in fat mass (p < 0.01 vs. baseline), a decrease in insulin resistance (p < 0.01 vs. six months; p = 0.01 vs. baseline) and a decrease in triglycerides (p < 0.01) were observed. Intima media thickness was greater in GH deficiency than in controls (p = 0.01) before therapy, and was unchanged after twelve months of therapy, whereas the flow-mediated dilatation tended to improve (p = 0.05). CONCLUSIONS: GH replacement is able to reverse typical metabolic and body composition alterations in patients with adult GH deficiency after twelve months, but it is unable to revert the vascular alteration completely. Flow-mediated dilatation seems to be a more precocious marker of the remission of arterial damage. PMID- 16477536 TI - Gastrointestinal inflammation and the circulating IGF system in humans. AB - Insulin-like growth factors (IGFs) and their binding proteins (IGFBPs) have important anabolic functions in normal tissue growth, which in excess may lead to tumorigenesis. In the present study, circulating IGF-I, IGF-II and their binding proteins (IGFBP-3, IGFBP-2 and IGFBP-1) were determined in 92 adult patients with gastrointestinal inflammation (Crohn's disease, colitis ulcerosa, gastritis, duodenitis errosiva, gastrointestinal candidiasis, and rotaviral and adenoviral enteritis). Serum IGF concentrations were measured by radioimmunoassay, while IGFBP profiles and IGFBP proteolytic patterns were characterized by immunoblotting. Concentrations of both IGF-I and IGF-II were significantly (p < 0.001) lower in patients than in healthy subjects. Immunoblotting demonstrated a decreased amount of intact IGFBP-3 (by approximately 60%), whereas IGFBP-2 and IGFBP-1 were increased (approximately 1.7 and 3.5-fold, respectively). No alteration in either fragmentation pattern or relative degree of proteolysis was detected in patients compared to the control group. It may be concluded that the IGF system is seriously imbalanced in patients with gastrointestinal inflammation, regardless of primary cause. These findings may help towards a better understanding of the metabolic outcome of the inflammatory process, and possibly in predicting the efficiency of patient recovery. PMID- 16477538 TI - Relationship between serum concentrations of interleukin-6 and tumor necrosis factor alpha in female Turkish subjects with normal and impaired glucose tolerance. AB - Serum interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNFalpha) concentrations were measured in subjects during two-hour glucose loading in order to investigate the effects of glucose on serum IL-6 and TNFalpha concentrations. Twenty-six female subjects (mean age 60 +/- 10 years) had normal glucose tolerance (NGT) and nineteen female subjects (mean age: 63 +/- 9 years) had impaired glucose tolerance (IGT) according to WHO criteria. Serum IL-6 and TNFalpha concentrations were measured by chemiluminescent immunometric assay. Subjects with IGT have higher fasting serum TNFalpha levels than subjects with NGT (p < 0.01). Serum IL-6 and TNFalpha concentrations were elevated during glucose loading (for each comparison, p < 0.01). The increase in serum TNFalpha concentrations in IGT was greater than in NGT (p < 0.01). Serum IL-6 and TNFalpha concentration significantly correlated with insulin and glucose in IGT group (for each comparison, p < 0.01). The correlation between serum glucose and cytokines concentrations was significant in IGT (for each comparison, p < 0.01). There was also a positive correlation between serum IL-6 and TNFalpha in NGT and IGT (for each comparison, p < 0.01). In conclusion, hyperglycemia is associated with increased circulating cytokine concentrations and fasting TNFalpha concentrations seem to be more associated with IGT than IL-6. PMID- 16477537 TI - Adipose tissue and circulating endothelial cell specific molecule-1 in human obesity. AB - Adipocytes produce the endothelial-cell specific molecule-1 (ESM-1), which inhibits leukocyte adhesion and migration through the endothelium. This study investigates ESM-1 expression and regulation in human adipose tissue. Subcutaneous abdominal adipose tissue was obtained from seventy postmenopausal women. Fourteen women subsequently underwent non-pharmacological weight reduction. In vitro experiments were performed on adipocytes isolated from human mammary adipose tissue. We determined gene expression by TaqMan RT-PCR and measured ESM-1 levels in serum and cell culture medium by ELISA. Mature adipocytes produced ESM-1. ESM-1 gene expression was higher in adipocytes than in preadipocytes. Cortisol inhibited ESM-1 gene expression in preadipocytes. Insulin and cortisol inhibited adipocyte ESM-1 production in adipocytes. This inhibitory effect of insulin was attenuated by insulin resistance, as ESM-1 gene expression in subcutaneous adipose tissue was increased in obese, hyperinsulinemic women. In contrast, ESM-1 serum levels were reduced in obese women and inversely correlated to C-reactive protein levels. Five percent weight loss did not markedly change gene expression. Circulating ESM-1 levels increased significantly, albeit modestly. ESM-1 is actively produced by adipocytes. However, since ESM-1 adipocyte gene expression and circulating plasma levels are not correlated, other sources of ESM-1 may be more important. Circulating ESM-1 levels are reduced in the overweight and obese, consistent with the notion that ESM-1 may play some role in obesity-associated vascular disease. PMID- 16477539 TI - Effects of coffee consumption on glucose tolerance, serum glucose and insulin levels--a cross-sectional analysis. AB - OBJECTIVE: Coffee has several metabolic effects that could reduce the risk of type 2 diabetes. Our objective was to examine the effects of coffee consumption on glucose tolerance, glucose and insulin levels. RESEARCH DESIGN AND METHODS: A subsample of subjects aged 45 to 64 years in 1987 and in 1992 from the population based FINRISK study (12,287 individuals) was invited to receive the standard oral glucose tolerance test at baseline. Plasma samples were taken after an overnight fast, and a two-hour oral glucose tolerance test was administered. Fasting and two-hour plasma glucose and insulin were measured in 2434 subjects with data on coffee use and potential confounders. RESULTS: After adjustment for potential confounding factors (age, body mass index, systolic blood pressure, occupational, commuting and leisure time physical activity, alcohol and tea drinking, smoking), coffee consumption was significantly and inversely associated with fasting glucose, two-hour plasma glucose, and fasting insulin in both men and women. Coffee consumption was significantly and inversely associated with impaired fasting glucose, impaired glucose regulation, and hyperinsulinemia among both men and women and with isolated impaired glucose tolerance among women. CONCLUSIONS: In this cross-sectional analysis, coffee showed positive effects on several glycemia markers. PMID- 16477540 TI - Prolonged ventricular repolarization measured by corrected QT interval (QTc) in subclinical hyperthyroidism. AB - Overt hyperthyroidism and hypothyroidism exert a major effect on cardiac function and on ECG. The influence of subclinical hyperthyroidism on the circulatory system is still under debate. Few studies examined the effect of thyroid hormones on ventricular repolarization measured by corrected QT interval (QTc). Longer QTc is associated with increased risk of arrhythmia and cardiac mortality. The aim of this study was to examine the influence of subclinical hyperthyroidism on ventricular repolarization measured by corrected QTc in a standard 12-lead electrocardiogram. The examined group consisted of thirty-two patients with subclinical hyperthyroidism; the controls were thirty-nine healthy individuals. In the group with subclinical hyperthyroidism, we observed a significant increase in heart rate (80.3 +/- 10.59 vs. 73.7 +/- 11.37 bpm, p < 0.05). The mean corrected QTc was 0.434 +/- 0.0207 seconds and 0.414 +/- 0.0208 in the examined groups and in controls, respectively (p < 0.001). QTc did not correlate with free thyroxin concentrations (p = 0.5084). CONCLUSION: Corrected QT intervals were significantly longer in patients with subclinical hyperthyroidism. PMID- 16477541 TI - Influence of thyroid disorders on kidney angiotensinase activity. AB - Thyroid disorders affect renal function, which involves changes in local renin angiotensin system (RAS). Angiotensin peptide levels in the tissue are regulated by the activity of several aminopeptidases (AP) known as angiotensinases. The nature and consequences of the thyroid-induced RAS changes are not completely understood. We investigated the relationship between thyroid status (hyper- and hypothyroidism) and several kidney AP actions involved in RAS control. We have determined fluorometrically soluble (SOL) and membrane-bound (M-B) alanylaminopeptidase (AlaAP), glutamylaminopeptidase (GluAP) and aspartylaminopeptidase (AspAP) activity using naphthylamide derivatives as substrates. Sprague-Dawley rats were divided into three groups--control, hyperthyroid, and hypothyroid. Hyperthyroidism was induced by daily subcutaneous injection of L-thyroxin (300 microg/kg/day). Hypothyroidism was induced by continuous administration of methimazole (0.03%) in drinking water. Hypothyroid animals demonstrated a significant increase in SOL and M-B GluAP activity in renal cortex and a decrease in M-B AlaAP compared to euthyroid rats. This result may suggest higher Ang III availability. In hyperthyroid animals, M-B AlaAP and M B AspAP activity increased significantly, which may suggest increased Ang III to Ang IV metabolism and greater formation of Ang 2-10, respectively. In contrast, no differences were observed between euthyroid and hypothyroid animals for SOL and M-B AP activity in renal medulla. However, hyperthyroid animals demonstrated a significant decrease in SOL and M-B GluAP activity compared to euthyroid rats, which may suggest a greater availability of Ang II in renal medulla. Alterations in angiotensin metabolism may, in part, account for some changes in renal function during thyroid disorders. PMID- 16477542 TI - Does autoantibody-negative Graves' disease exist? A second evaluation of the clinical diagnosis. AB - Advanced technical methods are essential for accurate diagnosis of Graves' or Basedow's disease (GD). Inadequate methods may lead to a false diagnostic conclusion. We have analyzed the clinical features and methodology aspects of cases diagnosed as GD with negative findings for TSH receptor autoantibodies. The initial diagnosis was based on clinical findings (patient record, hypermetabolic state, goiter palpation) and laboratory testing (fT4 and TSH). From a total of 255 newly registered patients with GD, fifty-one (20%) were negative in a conventional porcine TBII assay. All fifty-one patients were retested with 131I or 99mTc uptake tests, thyroid scintigraphy, and a second-generation TBII assay. Results disclosed twenty-one cases (8.3%) with diagnosis other than GD: ten cases of autonomous hyperthyroidism (Plummer's disease), seven cases of painless thyroiditis and four cases of euthyroid endocrine ophthalmopathy. All twenty-one patients remained negative in the second-generation TBII assay. Measurement by second-generation TBII assay was performed on the remaining thirty patients initially found negative for TBII. As a result of this reevaluation, only 234 of the original 255 patients had GD. Of those, 231 (204 according to porcine plus 27 according to human TRAb assay) had detectable TBII (98.7%). This investigation stresses the problem of correct diagnosis and the methodological limitations in the assessment of laboratory parameter validity in GD. Based on this work, TSH receptor autoantibody-negative GD is extremely rare. PMID- 16477543 TI - Absorption kinetics of levothyroxine is not altered by proton-pump inhibitor therapy. PMID- 16477545 TI - Doxycycline compared with benzathine penicillin for the treatment of early syphilis. AB - BACKGROUND: Doxycycline is the preferred recommended second-line agent for the treatment of syphilis, although efficacy data from controlled trials are lacking. We compared the serological responses of patients with early syphilis treated with doxycycline with the responses of patients treated with benzathine penicillin G (BPG). METHODS: All patients who received a diagnosis of early syphilis attending 2 public sexually transmitted disease clinics in Baltimore, Maryland, who were treated with doxycycline (100 mg orally, twice daily for 14 days) between October 1993 and June 2000 were eligible. Patients treated with BPG (a single dose of 2.4 million units intramuscularly) were selected as the control group. Inclusion criteria included a clinician-recorded diagnosis of primary, secondary, or early latent syphilis with reactive serological test results at the time of diagnosis and at least 1 follow-up serological test titer. Serological failure was defined as lack of a 4-fold drop in rapid plasma reagin titer 270-400 days after treatment, or a 4-fold increase in titer 30-400 days after therapy. RESULTS: During the study period, 1558 patients were treated for early syphilis, and 87 received doxycycline. Of those treated with doxycycline, 34 met the inclusion criteria. Seventy-three patients from a randomly selected group of 200 age-matched individuals treated with BPG met the inclusion criteria. There were 4 patients with serological failure in the BPG group (5.5%; 95% confidence interval [CI], 1.6%-13.8%) and 0 patients with serological failure in the doxycycline group (0%; 95% CI, 0%-10.3%; P=.2). The median times to successful serological responses for patients in the doxycycline and BPG groups were 106 days (95% CI, 75-149 days) and 137 days (95% CI, 111-172 days), respectively (P=.6). CONCLUSION: Doxycycline appears to be an effective agent for the treatment of early syphilis. PMID- 16477544 TI - Chromium chloride inhibits TNFalpha and IL-6 secretion in isolated human blood mononuclear cells exposed to high glucose. PMID- 16477546 TI - Genotyping and preemptive isolation to control an outbreak of vancomycin resistant Enterococcus faecium. AB - BACKGROUND: Control of vancomycin-resistant Enterococcus faecium (VRE) in European hospitals is hampered because of widespread asymptomatic carriage of VRE by healthy Europeans. In 2000, our hospital (The University Medical Center Utrecht, Utrecht, The Netherlands) was confronted with a large outbreak of VRE. INTERVENTION: On the basis of genotyping (by pulsed-field gel electrophoresis), epidemic and nonepidemic VRE strains were distinguished, and infection-control measures were exclusively targeted toward epidemic VRE. The outbreak was retrospectively divided into 3 periods of different infection-control measures. Compliance with use of alcohol-based hand rubs was enforced during all periods. Period I involved active surveillance, isolation of carriers, and cohorting (duration, 4 months); preemptive isolation of high-risk patients for VRE colonization was added in period II (7 months); and cohorting and preemptive isolation were abandoned in period III (18 months). METHODS: When the outbreak was identified, 27 patients in 6 wards were colonized; 93% were colonized with an epidemic VRE strain. Detection rates of nonepidemic VRE were 3.5%, 3.0%, and 2.9% among 683, 810, and 977 screened patients in periods I, II, and III, respectively, comparable to a prevalence of 2% (95% confidence interval [CI], 1% 3.5%) among 600 nonhospitalized persons. The relative risks of detecting epidemic VRE in periods II and III, compared with period I, were 0.67 (95% CI, 0.41-1.10) for period II and 0.02 (95% CI, 0.002-0.6) for period III. Infection-control measures were withheld for patients colonized with nonepidemic VRE (76 [54%] of 140 patients with a test result positive for VRE). Use of alcohol-based hand rubs increased by 31%-275% in outbreak wards. CONCLUSION: Genotyping-targeted infection control, isolation of VRE carriers, enhancement of hand-hygiene compliance, and preemptive isolation successfully controlled nosocomial spread of epidemic VRE infection. PMID- 16477547 TI - Outbreak of multidrug-resistant Salmonella enterica serotype Typhimurium Definitive Type 104 infection linked to commercial ground beef, northeastern United States, 2003-2004. AB - BACKGROUND: Multidrug-resistant Salmonella enterica serotype Typhimurium Definitive Type 104 (DT104) emerged in the 1990s and is associated with greater clinical severity than pansusceptible S. Typhimurium. Although infection with DT104 is common in the United States, it is rarely associated with outbreaks. From October to December 2003, a cluster of DT104 infections with indistinguishable pulsed-field gel electrophoresis patterns was identified in the northeastern United States. METHODS: A case-control study that assessed exposures compared case patients to age- and geography-matched control subjects. Information on consumer purchasing and grocery store suppliers was used to trace the implicated food to its source. RESULTS: We identified 58 case patients in 9 states by pulsed-field gel electrophoresis. Representative isolates were phage type DT104 and were resistant to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, and tetracycline (R-type ACSSuT). Of 27 patients interviewed for the case-control study, 41% were hospitalized (median duration of hospitalization, 4 days). Compared with 71 healthy control subjects, case patients had more medical comorbidities (matched odds ratio, 4.3; 95% confidence interval, 1.5-12.7). Illness was associated with consuming store-bought ground beef prepared as hamburgers at home (matched odds ratio, 5.3; 95% confidence interval, 1.9-15.3) and with eating raw ground beef (P< or =.001). Seven case patients (27%), but no control subjects, ate raw ground beef. Product traceback linked cases to a single large ground beef manufacturer previously implicated in a multistate outbreak of highly drug-resistant Salmonella enterica Newport infections in 2002. CONCLUSIONS: This first multistate outbreak of highly drug resistant S. Typhimurium DT104 infection associated with ground beef highlights the need for enhanced animal health surveillance and infection control, prudent use of antimicrobials for animals, improved pathogen reduction during processing, and better product tracking and consumer education. PMID- 16477548 TI - Breakthrough trichosporonosis in patients with hematologic malignancies receiving micafungin. AB - BACKGROUND: Micafungin is a newly approved antifungal agent in the echinocandin class that is active against Candida species and Aspergillus species. However, this agent has limited activity against a number of fungi, including Trichosporon species. We describe 4 patients who developed disseminated trichosporonosis during the use of micafungin. No cases of trichosporonosis had been seen in the 2 years prior to January 2003, when micafungin became available in our hospital. METHODS: We reviewed microbiological records of patients at Kameda General Hospital (Kamogawa City, Chiba, Japan) from 1 January 2002 to 31 July 2005, and identified 4 patients whose blood culture results were positive for Trichosporon species. RESULTS: Since January 2003, four patients--3 with acute myelocytic leukemia and 1 with myelodysplastic syndrome--developed disseminated trichosporonosis while receiving treatment with micafungin with or without amphotericin B. The initial 2 isolates were identified as Trichosporon beigelii, and the later 2 isolates were identified as Trichosporon asahii. All 4 patients received micafungin, and 2 also received amphotericin B concomitantly. Minimal inhibitory concentrations of micafungin were >16 microg/mL for the 2 isolates available for susceptibility testing. One patient with hematologic recovery (neutrophils >500 cells/mm3) showed elimination of the fungus after receiving treatment with voriconazole. However, the 3 other patients without hematologic or immunological recovery died of disseminated infection. CONCLUSIONS: The rarity of trichosporonosis in our hospital and its emergence after the introduction of micafungin therapy support the idea that micafungin may exert a significant, selective pressure toward resistant fungi, such as Trichosporon species. Therefore, care should be taken regarding the possibility of trichosporonosis in patients receiving micafungin with or without amphotericin B. PMID- 16477549 TI - Management and outcomes of a first recurrence of Clostridium difficile-associated disease in Quebec, Canada. AB - BACKGROUND: During an epidemic of Clostridium difficile-associated disease (CDAD) caused by a strain that is a hyper-producer of toxins A and B, the frequency of a first recurrence after metronidazole treatment of the initial episode doubled in 2003-2004, compared with 1991-2002. METHODS: To examine whether administration of metronidazole as treatment for a first recurrence of CDAD remained appropriate, we reviewed data for patients with CDAD diagnosed in a hospital in Quebec, Canada, during 1991-2005, who experienced a first recurrence. The frequency of a second recurrence within 60 days after the first one was measured using Kaplan Meier analysis. Cox regression was used for multivariate analysis. RESULTS: A total of 463 patients had a first recurrence of CDAD, of whom 154 (33.3%) experienced a second recurrence. Independent predictors of a second recurrence were age and duration of hospitalization after the first recurrence; this latter finding suggested that many such episodes were reinfections rather than relapses. Neither choice of treatment drug (metronidazole or vancomycin) nor use of the same drug for treatment of first recurrence, as had been used during the initial episode, was associated with increased risk of a second recurrence. However, 51 patients (11.0%) developed at least 1 complication (i.e., shock, need for colectomy, megacolon, perforation, or death within 30 days) during the first recurrence. Older age, a high leukocyte count, and renal failure at first recurrence were strongly associated with a complicated CDAD. CONCLUSIONS: Metronidazole is not inferior to vancomycin for treatment of patients with a first recurrence of CDAD, but the risk of complications with any treatment of CDAD may be higher than has previously been documented. PMID- 16477550 TI - Treatment of first recurrences of Clostridium difficile-associated disease: waiting for new treatment options. PMID- 16477551 TI - Effectiveness of education and an antibiotic-control program in a tertiary care hospital in Thailand. AB - BACKGROUND: We conducted a study to evaluate the impact of education and an antibiotic-control program on antibiotic-prescribing practices, antibiotic consumption, antimicrobial resistance, and cost of antibiotics in a tertiary care hospital in Thailand. METHODS: A study of the year before and the year after the intervention was performed. Inpatient antibiotic prescriptions were prospectively observed. Demographic characteristics, hospital unit, indication for antibiotic prescription, appropriateness of antibiotic use, reasons for inappropriate antibiotic use, antibiotic consumption (i.e., the rate of antibiotic use), bacterial resistance, and antibiotic cost data were collected. Interventions included education, introduction of an antibiogram, use of antibiotic prescription forms, and prescribing controls. RESULTS: After the intervention, there was a 24% reduction in the rate of antibiotic prescription (640 vs. 400 prescriptions/1000 admissions; P<.001). The incidence of inappropriate antibiotic use was significantly reduced (42% vs. 20%; P<.001). A sustained reduction in antibiotic use was observed (R2=0.692; P<.001). Rates of use of third-generation cephalosporins (31 vs. 18 defined daily doses [DDDs]/1000 patient-days; P<.001) and glycopeptides (3.2 vs. 2.4 DDDs/1000 patient-days; P=.002) were significantly reduced. Rates of use of cefazolin (3.5 vs. 8.2 DDDs/1000 patient-days; P<.001) and fluoroquinolones (0.68 vs. 1.15 DDDs/1000 patient-days; P<.001) increased. There were no significant changes for other antibiotic classes. Significant reductions in the incidence of infections due to methicillin-resistant Staphylococcus aureus (48% vs. 33.5%; P<.001), extended-spectrum beta -lactamase producing Escherichia coli (33% vs. 21%; P<.001), extended-spectrum beta lactamase-producing Klebsiella pneumoniae (30% vs. 20%; P<.001), and third generation cephalosporin-resistant Acinetobacter baumanii (27% vs. 19%; P<.001) were also observed. Total costs saving were USD 32,231 during the study period. CONCLUSIONS: Education and an antibiotic-control program constituted an effective and cost-saving strategy to optimize antibiotic use in a tertiary care center in Thailand. PMID- 16477552 TI - Controlling antibiotic use and resistance. PMID- 16477553 TI - Fluoroquinolone use and methicillin-resistant Staphylococcus aureus isolation rates in hospitalized patients: a quasi experimental study. AB - BACKGROUND: We investigated the possible association between fluoroquinolone use and the rate of methicillin-resistant Staphylococcus aureus (MRSA) recovery from consecutive hospitalized patients. METHODS: We conducted a nonrandomized, prospective, controlled interventional "fluoroquinolone-free" study at 4 large teaching hospitals in northwest France, catering to a total of 5,882,600 persons. During the intervention period (January through December 2001), fluoroquinolone use was prohibited at 1 of the 4 hospitals (Caen Hospital), unless no effective alternative was available. Three university hospitals were used as controls because they had similar preintervention rates of MRSA. RESULTS: During the intervention period (2001), the annual rate of fluoroquinolone use decreased from 54 to 5 defined daily doses per 1000 patients per day at Caen Hospital and remained stable in the control hospitals. At the end of the intervention, the rate of MRSA isolation was significantly lower at Caen Hospital than at the control hospitals (353 [32.3%] of 1093 S. aureus isolates were MRSA, compared with 2495 [36.8%] of 6787 isolates; odds ratio, 0.82; 95% confidence interval, 0.69-0.99; P=.036), as determined on the basis of a marginal model that took into account within-hospital clustering. In a before-after time series analysis, compared with forecasted rates, there was a significant downward trend in observed monthly rates of MRSA isolation at Caen Hospital at the end of the intervention. CONCLUSION: This quasi experimental study confirms the association between fluoroquinolone use and MRSA isolation among hospitalized patients. PMID- 16477554 TI - Infection control or formulary control: what is the best tool to reduce nosocomial infections due to methicillin-resistant Staphylococcus aureus? PMID- 16477555 TI - The impact of penicillin resistance on short-term mortality in hospitalized adults with pneumococcal pneumonia: a systematic review and meta-analysis. AB - BACKGROUND: The clinical impact of penicillin resistance on the outcome of pneumococcal pneumonia has remained controversial. We performed a meta-analysis of prospective cohort studies to examine the association between penicillin resistance and short-term all-cause mortality for pneumococcal pneumonia. METHODS: We retrieved studies published in any language by a comprehensive search of the Medline, Current Contents, and Embase databases for all appropriate articles published up to January 2005. We also manually reviewed bibliographies of retrieved articles, recent national treatment guidelines, and review articles. We included prospective cohort studies that involved adult subjects, and we examined the association between penicillin resistance and short-term mortality for pneumococcal pneumonia. Two reviewers independently extracted data on crude and adjusted risk estimates of all-cause mortality for pneumococcal infections with different levels of penicillin resistance and assessed the methodological quality of selected studies. We also contacted authors to obtain additional information. We performed meta-analyses using a random-effect model. RESULTS: Of 1152 articles identified in the search, 10 studies that involved 3430 patients (most of whom were hospitalized) were included. The mortality rate was 19.4% in the penicillin-nonsusceptible Streptococcus pneumoniae group and 15.7% in the penicillin-susceptible S. pneumoniae group. The combined relative risks of all cause mortality for the penicillin-nonsusceptible, -intermediate, and -resistant S. pneumoniae groups, compared with the penicillin-susceptible S. pneumoniae group, were 1.31 (95% confidence interval [CI], 1.08-1.59), 1.34 (95% CI, 1.13 1.60), and 1.29 (95% CI, 1.01-1.66), respectively. The combined adjusted relative risks of mortality for penicillin-nonsusceptible versus penicillin-susceptible S. pneumoniae group was 1.29 (95% CI, 1.04-1.59) for the 6 studies that adjusted for age, comorbidities, and severity of illness. There was minimal between-study heterogeneity in these analyses. CONCLUSION: Penicillin resistance is associated with a higher mortality rate than is penicillin susceptibility in hospitalized patients with pneumococcal pneumonia. Additional efforts are needed to understand the mechanisms of this association. PMID- 16477556 TI - The clinical relevance of penicillin-resistant Streptococcus pneumoniae: a new perspective. PMID- 16477557 TI - Development of a genetic assay to distinguish between Leishmania viannia species on the basis of isoenzyme differences. AB - BACKGROUND: Tegumentary leishmaniasis in Latin America is caused mainly by Leishmania viannia braziliensis complex parasites. L. braziliensis and Leishmania viannia peruviana are the 2 predominant Leishmania species in Peru. L. braziliensis is more virulent, because it can cause mucocutaneous leishmaniasis, known as espundia, that results in severe facial destruction. Early identification of the species that causes the initial cutaneous infection would greatly help to prevent mucocutaneous leishmaniasis, because it would allow more aggressive treatment and follow-up. However, because of the close genetic similarity of L. braziliensis and L. peruviana, there currently exists no simple assay to distinguish between these species. METHODS: We cloned the mannose phosphate isomerase gene from both L. braziliensis and L. peruviana. It is the only known isoenzyme capable of differentiating between L. braziliensis and L. peruviana in multilocus enzyme electrophoresis. Interestingly, only a single nucleotide polymorphism was found between the mannose phosphate isomerase genes from L. braziliensis and L. peruviana, resulting in an amino acid change from threonine to arginine at amino acid 361. A polymerase chain reaction assay was developed to distinguish the single nucleotide polymorphism of the mannose phosphate isomerase gene to allow for the specific identification of L. braziliensis or L. peruviana. RESULTS: This assay was validated with 31 reference strains that were previously typed by multilocus enzyme electrophoresis, successfully applied to patient biopsy samples, and adapted to a real-time polymerase chain reaction assay. CONCLUSIONS: This innovative approach combines new genetic knowledge with traditional biochemical fundamentals of multilocus enzyme electrophoresis to better manage leishmaniasis in Latin America. PMID- 16477558 TI - A tale of 2 alpha-herpesviruses: lessons for vaccinologists. AB - Of the 8 known herpesviruses that affect human beings, we only have successful vaccines against varicella zoster virus. This brief review compares the pathogenesis of varicella zoster virus with that of the closely related alpha herpesviruses herpes simplex virus 1 and 2, for which we have no satisfactory vaccines. The main objective of this review is to learn lessons from the success of varicella zoster virus vaccine that could be exploited for the development of successful vaccines against herpes simplex virus and perhaps against other herpes viruses. PMID- 16477559 TI - Endocarditis after acute Q fever in patients with previously undiagnosed valvulopathies. AB - We describe 3 cases of endocarditis after acute Q fever in 3 patients with clinically silent, undiagnosed valvulopathies, including mitral valve prolapse, minimal valvular leak, and biscuspid aortic valve. We conclude that, to prevent endocarditis, these minor valvulopathies must be actively searched for with echocardiography after diagnosis of acute Q fever. PMID- 16477560 TI - Hospitalizations for endemic mycoses: a population-based national study. AB - We performed a retrospective cohort study, using the 2002 Nationwide Inpatient Sample, a national database of hospital inpatient stays, to describe the incidence and epidemiology of endemic mycoses requiring hospitalization. An estimated 332 pediatric and 6003 adult patients with endemic mycoses required hospitalization (4.6 and 28.7 cases per 1 million children and adults, respectively). Crude mortality rates were 5% and 7% among children and adults, respectively. PMID- 16477561 TI - Preventing transmission of multidrug-resistant bacteria in health care settings: a tale of 2 guidelines. AB - Two guidelines for the control of multidrug-resistant organisms in health care facilities have appeared during the past 3 years--one from the Society for Healthcare Epidemiology of America and one, in draft form, from the Healthcare Infection Control Practices Advisory Committee of the Centers for Disease Control and Prevention. These guidelines reflect universal concern in the infection control community about today's unprecedented levels of activity of multidrug resistant organisms and about inadequate or inconsistent application of potentially effective control measures. The 2 guidelines provide detailed reviews of pertinent issues and evidence-based, rated recommendations, which overlap considerably. Recommendations regarding indications for active surveillance cultures and the extent of their use constitute the major divergence. Although implementation of comprehensive control plans for multidrug-resistant organisms advocated by both guidelines will require health care facilities to confront difficult programmatic issues, aggressive and widespread adoption of control measures for multidrug-resistant organisms is urgently needed. PMID- 16477562 TI - Nutrition and HIV infection: review of weight loss and wasting in the era of highly active antiretroviral therapy from the nutrition for healthy living cohort. AB - Despite major advances in the treatment and survival of patients infected with human immunodeficiency virus (HIV), weight loss and wasting remain common problems. In the HIV-infected population, weight loss is associated with lower CD4+ cell counts and is an independent predictor of mortality. The etiology of weight loss and wasting is complex and multifactorial. We discuss, on the basis of a large longitudinal cohort that examined nutritional status in HIV infection, data on weight loss and wasting from the present clinical era. The definition, prevalence, and significance of HIV-associated weight loss and wasting are summarized. The etiology of weight loss is discussed for 2 main categories: inadequate nutrient intake and altered metabolism. Finally, studies of interventions to treat HIV-associated weight loss and wasting are discussed. This information is intended to raise awareness among health care providers of HIV infected patients that weight loss and wasting remain important acquired immunodeficiency syndrome-defining conditions, despite the advent of HAART. PMID- 16477564 TI - Are all HIV type 1 strains created equal? PMID- 16477566 TI - Long-term effects of highly active antiretroviral therapy in pretreated, vertically HIV type 1-infected children: 6 years of follow-up. AB - BACKGROUND: Several studies of children with human immunodeficiency virus (HIV) type 1 infection have demonstrated sustained increases in CD4+ cell count, even when virological failure has occurred after receipt of highly active antiretroviral therapy (HAART), but these studies were of limited duration. Moreover, the CD4+ cell count threshold at which antiretroviral treatment should be initiated is still unsettled. The aim of this study was to define the long term impact of HAART on CD4+ cell percentage and viral load according to CD4+ cell percentages before HAART was initiated. METHODS: We conducted a retrospective study of 113 pretreated HIV-1-infected children stratified by pre HAART CD4+ cell percentage (<5%, 5%-15%, 15%-25%, and >25%). The inclusion criteria were as follows: initiating HAART with a protease inhibitor, having 6 years of follow-up after starting HAART, having a CD4+ cell count or viral load recorded before initiation of HAART, and having received mono- or dual-nucleoside therapy before starting HAART. RESULTS: During the first 2 years of HAART, HIV-1 infected children experienced a significant increase in CD4+ cell percentage and a decrease in viral load (P<.05). During their last 4 years of receiving HAART, we found a significant decrease in viral load but not an increase in CD4+ cell percentage, because the CD4+ cell percentage reached a plateau after the second year of HAART. Moreover, children with CD4+ cell percentages of <5% at baseline did not achieve CD4+ cell percentages of >25% after 6 years of HAART. Children with CD4+ cell percentages of 5%-25% at baseline had a strong negative association with achieving CD4+ cell percentages of >30% for at least 6 and 12 months but not with achieving CD4+ cell percentages of >30% for at least 24 months. CONCLUSIONS: Long-term HAART allowed for restoration of CD4+ cell counts and control of viral loads in HIV-1-infected children. However, initiating HAART after severe immunosuppression has occurred is detrimental for the restoration of the CD4+ cell count. PMID- 16477565 TI - Effect of HIV infection on atypical squamous cells of undetermined significance. AB - BACKGROUND: Detection of atypical squamous cells of undetermined significance (ASCUS) is a cervical cytologic finding that is suggestive but not definitive of squamous intraepithelial lesions (SILs). METHODS: We examined the risk, characteristics, and progression of ASCUS in women with and without human immunodeficiency virus (HIV) infection. Cervical Papanicolou (Pap) test and colposcopy data were obtained at the first 10 semiannual visits for the HIV Epidemiology Research study of 774 HIV-infected and 480 demographically similar, HIV-uninfected women in the United States. Multiple logistic regression models and Cox proportional hazards models were utilized. RESULTS: ASCUS was more common among HIV-infected women (odds ratio [OR], 1.6 [95% confidence interval {CI}, 1.3 2.0] to 2.6 [95% CI, 1.9-3.6]) after adjustment for human papillomavirus (HPV) infection and other risk factors (e.g., race, condyloma, and prior Pap test result). Among women with normal Pap test results at enrollment, the cumulative incidence of ASCUS was 78% among HIV-infected women and 38% among HIV-uninfected women. HIV-infected and HIV-uninfected women with ASCUS did not differ by prevalence of indices of inflammation (inflammation on Pap test and leukocytes on cervical gram stain). HPV infection, including high risk types, was more common among HIV-infected women with ASCUS. Among women with ASCUS, 60% of HIV-infected and 25% of HIV-uninfected women developed SILs (P < .01). Compared with HIV infected women with higher CD4+ lymphocyte counts, HIV-infected women with CD4+ lymphocyte counts < 200 cells/microL were more likely to present subsequently with a SIL detected by Pap test (OR, 1.7; 95% CI, 0.8-3.6). CONCLUSIONS: Higher risk of SIL following the appearance of ASCUS among HIV-infected women, especially women with low CD4+ lymphocyte counts, supports the need for follow up with colposcopy and histologic examination, as indicated, to allow early detection and treatment of SIL. PMID- 16477567 TI - Week-12 response to therapy as a predictor of week 24, 48, and 96 outcome in patients receiving the HIV fusion inhibitor enfuvirtide in the T-20 versus Optimized Regimen Only (TORO) trials. AB - BACKGROUND: Early virological response to antiretroviral therapy is predictive of long-term treatment outcome in therapy-naive patients. In treatment-experienced patients, such correlations are less well defined, because initial responses may be less pronounced and transient because of accumulated cross-resistance to prior therapies. Our objectives were to explore how the virological and immunological status of treatment-experienced patients at an early time point (week 12) during enfuvirtide-based therapy predicted their responses at weeks 24, 48, and 96 in the T-20 versus Optimized Regimen Only (TORO) trials. METHODS: Post hoc, modified, on-treatment and intent-to-treat analyses were performed to determine whether the relationship between virological and immunological outcomes at weeks 24, 48, and 96 were predicted by the patients' week-12 responses to therapy. RESULTS: Using a modified on-treatment analysis for patients who, by week 12, achieved a decrease in their HIV-1 RNA load of > or =1 log10 copies/mL, 39.2% (95% CI, 33.6%-44.8%) and 59.5% (95% CI, 53.8%-65.1%) achieved a viral load of <50 copies/mL or <400 copies/mL at week 96, respectively, compared with 1.3% (95% CI, 0%-3.8%) and 2.6% (95% CI, 0%-6.1%) of patients, respectively, who did not achieve an early virological response. Using the same modified on-treatment analysis method for patients who, at week 12, achieved a CD4 cell count increase of > or =50 cells/mm3, 87.2% (95% CI, 82.6-91.8) maintained or improved this response through week 96, compared with 56.6% (95% CI, 47.5-65.8) of patients who did not achieve this early categorical immunological response. CONCLUSION: Enfuvirtide-based treatment regimens are associated with a rapid and durable response. Week-12 virological and immunological responses to treatment with enfuvirtide are predictive of subsequent outcomes in triple-class treatment experienced patients. PMID- 16477568 TI - Effect of maintaining highly active antiretroviral therapy on AIDS events among patients with late-stage HIV infection and inadequate response to therapy. AB - BACKGROUND: We evaluated the effect of maintaining highly active antiretroviral therapy (HAART) on the development of new acquired immunodeficiency syndrome (AIDS)-related events in patients with late-stage human immunodeficiency virus (HIV) infection who had suboptimal CD4+ cell count and viral load responses to HIV therapy. METHODS: In patients with pretreatment CD4+ cell counts of <200 cells/mm3, incidence rates of new AIDS-related events occurring during HIV treatment were calculated during period 1 (pre-HAART era, 1990-1995; 88 patients) and period 2 (HAART era, 1996-2004; 214 patients) according to CD4+ cell count responses while receiving treatment. Cox multivariate model was used to compare rates of AIDS-related events from period 2 with those from period 1 according to specific CD4+ cell count response categories and rates of AIDS-related events for various viral load ranges within CD4+ cell count categories during period 2. RESULTS: For period 2 patients with CD4+ cell counts <50 cells/mm3 and viral loads >100,000 copies/mL, the rate of AIDS-related events (39.3 events per 100 person-years) was significantly lower than that for period 1 patients with CD4+ cell counts <50 cells/mm3 (76.4 events per 100 person-years; P=.02). This held true for patients with CD4+ cell counts <100 cells/mm3; there were also significantly fewer AIDS-related events in period 2 (18 events per 100 person years) than in period 1 (65.2 events per 100 person-years; P=.001), including those events occurring among period 2 patients with viral loads >100,000 copies/mL (29.5 events per 100 person-years; P=.01). Similarly, for patients with CD4+ cell counts of 100-200 cells/mm3, there were fewer AIDS-related events in period 2 (7.8 events per 100 person-years) than in period 1 (34.5 events per 100 person-years; P=.001); even for patients in period 2 with viral loads >100,000 copies/mL (15.4 events per 100 person-years; P=.04). CONCLUSIONS: Our data suggest that, even among patients with late-stage HIV infection and inadequate CD4+ cell count and viral load responses to HIV therapy, maintaining HAART may reduce the incidence of AIDS-related events. PMID- 16477569 TI - Detection of immunoglobulin M antibody to hepatitis A virus in patients without acute hepatitis A: the usefulness of specific immunoglobulin G avidity. PMID- 16477570 TI - Where is the humility for the limitations of research? PMID- 16477563 TI - Different rates of disease progression of HIV type 1 infection in Tanzania based on infecting subtype. AB - BACKGROUND: Many different subtypes of human immunodeficiency virus (HIV) type 1 have been identified, particularly in sub-Saharan Africa. However, much remains unknown regarding the relative pathogenicity of these subtypes and their influence on the clinical progression of HIV infection. We examined prospectively the associations between HIV-1 subtypes A, C, and D and recombinant viruses, as well as the rates of disease progression in a cohort of seropositive women from Dar es Salaam, Tanzania. METHODS: A total of 428 pregnant mothers participating in a larger controlled trial of the effect of vitamin supplements were selected for DNA sequencing of their HIV-1 subtype. Plasma viral load was measured at baseline, and CD4+ cell counts was assessed at baseline and at regular intervals throughout the follow-up period. Proportional hazards regression (hazards ratio [HR]) analysis was used to measure the association between viral subtype and the rate of disease progression. RESULTS: Relative to patients with subtype A, patients with subtype D experienced the most rapid progression to death (HR, 2.27; 95% confidence interval [CI], 1.46-3.52) or to the World Health Organization stage 4 of illness (HR, 1.94; 95% CI, 1.20-3.14) and to a CD4+ cell count of <200 cells/mm3 (HR, 2.12; 95% CI, 1.42-3.17). After adjustment for viral load, CD4+ cell count, and other baseline covariates, the associations remained similar. CONCLUSIONS: We observed heterogeneity in the rates of disease progression of HIV-1 disease in infected persons, on the basis of the infecting subtype. Subtype D was associated with the most rapid progression of the disease, relative to the other 3 categories of viruses in our cohort. PMID- 16477572 TI - Unusual pathogens and multidrug-resistant bacteria in tsunami survivors. PMID- 16477574 TI - A case of Bergeyella zoohelcum bacteremia after ingestion of a dish prepared with goat blood. PMID- 16477576 TI - Rifampin and pyrazinamide for treatment of latent tuberculosis infection. PMID- 16477577 TI - Paradoxical reaction during tuberculosis treatment in HIV-seronegative patients. PMID- 16477578 TI - Glucose homeostasis abnormalities and gatifloxacin. PMID- 16477580 TI - The validity and reliability of the Turkish version of Alzheimer's Disease Assessment Scale-Cognitive Subscale (ADAS-Cog) in patients with mild and moderate Alzheimer's disease and normal subjects. AB - OBJECTIVES: The cognitive subscale of the Alzheimer's Disease Assesment Scale (ADAS-Cog) is the most widely used test in clinical trials dealing with Alzheimer's disease (AD). The aim of this study was to investigate the validity and reliability of the Turkish version of ADAS-Cog. METHODS: Twenty-nine patients with AD, fulfilling NINCDS-ADRDA criteria of probable AD, who were in stage 3-5 according to the Global Deterioration Scale (GDS), and 27 non-demented control subjects with similar age, gender and educational status were recruited for the study. The Turkish version of ADAS-Cog, Standardized Mini Mental Status Examination (MMSE) and Short Orientation-Memory-Concentration Test (SOMCT) were applied to both of the groups. Inter-rater reliability, internal consistency, test-retest reliability; face validity, differential validity and convergent validity were statistically analyzed. RESULTS: Both MMSE and ADAS-Cog have significantly differentiated patients with AD and control subjects (p < 0.001). A significant correlation was established between MMSE and ADAS-Cog scores in AD group (r: -0.739). ADAS-Cog was also highly significantly correlated with GDS (r: 0.720) and SOMCT (r: 0.738). For the group with AD, control and whole cohort coefficients of internal consistency, Cronbach's alpha: 0.800, 0.515, 0.873 were found respectively. Inter-rater reliability for total ADAS-Cog score was found as ICC: 0.99 and 0.98 and test-retest reliability was found as ICC: 0.91 and 0.95 for demented and nondemented subjects, respectively. CONCLUSION: The Turkish version of ADAS-Cog has been found to be highly reliable and valid in differentiating patients with mild and moderate AD from nondemented subjects. PMID- 16477579 TI - Resveratrol is efficiently glucuronidated by UDP-glucuronosyltransferases in the human gastrointestinal tract and in Caco-2 cells. AB - Resveratrol (3, 5, 4'-trihydroxy-trans-stilbene), a natural polyphenol present in grapes and peanuts, has been reported to exert a variety of potentially therapeutic effects. The aim of this study was to determine the contribution of the gastrointestinal (GI) tract to the glucuronidation of this compound and its cis-isomer, which also occurs naturally. For this purpose, glucuronidation of the two resveratrol isomers was investigated in human microsomes prepared from: stomach, duodenum, four segments of the remaining small intestine (S-1 to S-4) and colon, and from the human intestinal cell lines Caco-2 and PD-7. cis- and trans-Resveratrol were efficiently glucuronidated in the GI tract with the formation of both 3-O- and 4'-O-glucuronides, however, the two stereoisomers were glucuronidated at different rates depending on the donor and the segment considered. Microsomes prepared from Caco-2 and PD-7 cells also efficiently glucuronidated cis-resveratrol and, to a lesser extent, the trans-isomer, however, only the 3-O-glucuronide was formed. Among the UDP glucuronosyltransferases (UGT) that are known to be expressed in the GI tract, the isoforms UGT1A1, 1A6, 1A8, 1A9 and 1A10 were active in glucuronidating trans- and/or cis-resveratrol. The results demonstrate that the GI tract may contribute significantly to the first pass metabolism of these naturally occurring polyphenols. PMID- 16477581 TI - Idiopathic myelofibrosis: pathogenesis to treatment. AB - Idiopathic myelofibrosis (IMF) is the least common of the chronic myeloproliferative disorders and carries the worst prognosis with a median survival of 4 years. It is a clonal haematopoietic stem-cell disorder and, although the pathogenesis remains unclear, approximately 50% of cases are known to possess an activating JAK2 V617F mutation. In contrast, the characteristic stromal proliferation is a reactive, or secondary, event that results from the aberrant release of a variety of growth factors from megakaryocytes and monocytes. Treatment for most cases is supportive, although androgens, recombinant erythropoietin, steroids and thalidomide are effective modalities for the amelioration of anaemia. Myelosuppression, splenectomy and irradiation are valuable therapeutic modalities for specific clinical situations. Prognostic scores are available to aid the identification of cases for whom bone marrow transplantation should be considered. Recently, the use of reduced intensity conditioning has resulted in prolonged survival and lower transplant-related mortality. This review summarises the recent advances in the disease's pathogenesis and discusses the role of the various therapeutic options. PMID- 16477582 TI - Nursing home institutionalization: a source of eustress or distress for the elderly? AB - OBJECTIVE: To investigate why elderly persons moved to a nursing home and detect any change in their psychopathological and cognitive profile, dependency and perceived QOL. METHOD: We considered 100 elderly consecutively admitted to a nursing home, within 1 week of admission and 6 months later. Data were collected from medical and nursing records (medical and psychiatric history, ADL), interviews (MMSE, reasons for admission) and self-report instruments (GDS, BSI, WHOQOL-brief). RESULTS: At admission, 68 subjects were able to participate in the survey and agreed to take part. Only 5.9% (n = 4) had moved to the nursing home by their own choice, 26.5% (n = 18) because they were lonely and 36.7% (n = 25) because they had no caregiver available. The reasons for admission reported by the interviewed residents only partly corresponded with the ones noted in their records. Twenty-six subjects had an MMSE score >or= 18 and agreed to undertake complete assessment. The scores of 20 of the 26 subjects exceeded the cut-off on the GDS and five of the nine BSI subscales. During the follow-up period, 19 of the 68 originally assessed residents died (33 of the entire sample of 100) and one refused to continue to take part in the survey. The mean MMSE and ADL scores of the 48 survivors decreased from 16.87 (SD +/- 7.32) to 14.27 (SD +/- 7.24; t = 4.89, p < 0.000) and from 8.22 (SD +/- 3.55) to 6.39 (SD +/- 3.56; t = 5.34, p < 0.000), respectively. The scores achieved on GDI and BSI subscales worsened in 20 survivors with MMSE >or= 18. The score achieved on the QOL physical health domain also worsened. Comparison of the subjects who died and survived during the follow up period showed a significant difference in terms of ADL (6.60 SD +/- 4.221 vs 3.64 SD +/- 3.773 F = 11.639; p < 0.001). CONCLUSION: The 68 original participants expressed feelings of loneliness and marginalization, but these experiences were not noted in their personal records. Most subjects presented psychiatric symptoms and cognitive decline. After 6 months, the clinical condition of the sample, particularly the females, had worsened, with a 33% mortality rate. This suggests that moving to a nursing home did not bring about improvement or stabilization; rather, psychiatric symptoms worsened and quality of life was perceived more poorly. PMID- 16477583 TI - Validation of the Spanish version of the Addenbrooke's Cognitive Examination in a rural community in Spain. AB - BACKGROUND: The Addenbrooke's Cognitive Examination (ACE) is a brief cognitive test battery designed to detect and differentiate Alzheimer's disease (AD) and frontotemporal dementia (FTD). Translations of this instrument into French and Malayalam have been recently published OBJECTIVE: To adapt and validate the ACE into Spanish in a rural population of low-educational level. SUBJECTS: A clinical group, composed of 70 patients affected by dementia and 25 patients with memory complaints without dementia, was compared with 72 controls matched for gender, age and educational level METHOD: The clinical group was studied with standard neuropsychological instruments, all patients underwent neuroimaging [Computerized Tomography (CT) or Magnetic Resonance Imaging (MRI), and Single Photon Emission Tomography (SPECT) in all cases of suspected FTD], as well as routine neurological examination. Both groups were studied with the ACE and Clinical Dementia Rating scale (CDR). Sensitivity, specificity, area under curve, reliability and Verbal-Language/ Orientation-Memory (VLOM) ratio were calculated. Subsequently, the sample was stratified regarding educational level in two groups. Receiver Operating Characteristics (ROC) curves were calculated for these conditions. Different cut-off points were calculated addressing educational level. RESULTS: ROC curves demonstrated the superiority of the ACE in the sub sample of patients that finished school at over 14 years old. VLOM ratio confirmed its usefulness for differential diagnosis between AD and FTD CONCLUSIONS: The Spanish version of the ACE is a useful instrument for dementia diagnosis. In our sample VLOM ratio results were useful for differential diagnosis between AD and FTD. Different cut-off points must be used for different educational levels. PMID- 16477584 TI - Association of depression with Alzheimer's disease and vascular dementia in an elderly Arab population of Wadi-Ara, Israel. AB - OBJECTIVES: Because dementia and depression share common risk factors, we investigated risk factors for depression in Arab subjects with Alzheimer's disease (AD) and vascular dementia (VaD). METHODS: In a cross-sectional population-based study, we conducted a door-to-door survey of all adults over age 60 in an Arab community of rural Israel. We conducted interviews, gave questionnaires, and collected DNA blood specimens for determination of ApoE genotype. RESULTS: Of the 823 individuals in this naturalistic sample, 168 had dementia of Alzheimer's type (DAT) and 49 had VaD. Vascular risk factors, including the ApoE-epsilon4 allele, were more prevalent among VaD than DAT subjects. Depressive symptoms were present in 57% of DAT patients and 86% of VaD patients. Depressed DAT individuals had a greater history of ischemic cardiovascular or cerebrovascular (CV/CBV) disease than non-depressed DAT subjects, but depressed DAT subjects were less likely to have the ApoE-epsilon4 allele. Within the VaD group, there was no difference in the distribution of cardiovascular risk factors in individuals with and without depressive symptoms, and ApoE-epsilon4 was more prevalent among subjects with depressive symptoms. CONCLUSIONS: Depressive symptomatology is prevalent among subjects with dementias in this Arab community. History of CV/CBV is associated with the presence of depressive symptoms in DAT. Further studies are needed to clarify the role of ApoE in depression onset in different ethnic groups with DAT. PMID- 16477586 TI - Effect of antidepressant use on admissions to hospital among elderly bipolar patients. AB - OBJECTIVE: The goal of this study was to examine the association between antidepressant use and hospitalization rate for mania or bipolar depression in a large, community-based sample of elderly bipolar patients. METHOD: Population based retrospective cohort design. Administrative healthcare databases were linked for all individuals aged 66 years or older in the Province of Ontario, Canada. Bipolar subjects who received a prescription for an antidepressant medication (n = 1,072) between 1 April 1997 and 31 March 2001 comprised the antidepressant cohort. The control group (n = 3,000) consisted of randomly selected subjects from the eligible bipolar population who did not receive a prescription for an antidepressant medication during the same surveillance period. Primary outcomes were admission to hospital for a manic/mixed or depressive episode. RESULTS: During a total of 5135 person-years of follow up, 113 admissions for a manic/mixed episode and 28 admissions for a depressive episode were identified. Model based estimates adjusted for a number of covariates revealed that, as compared with the control group, the antidepressant cohort had significantly lower likelihood of admissions for manic/mixed (adjusted rate ratio [aRR] = 0.5, 95% CI = 0.3-0.8) but not depressive episodes (aRR = 0.7, 95% CI = 0.2-2.2). CONCLUSION: Antidepressant use among elderly bipolar patients was associated with decreased rates of hospitalization for manic/mixed episodes. This finding requires confirmation with further data of antidepressant use among elderly bipolars. PMID- 16477585 TI - Depressive symptom profile in Parkinson's disease: a comparison with depression in elderly patients without Parkinson's disease. AB - OBJECTIVE: Depression is a common neuropsychiatric syndrome in Parkinson's disease (PD), and may be etiologically related to the neurochemical changes accompanying this disease. It is still unclear whether the disturbances of neurotransmitter activities lead to a specific profile of depressive symptoms, that is characteristic for PD and differs from that in depressed patients without PD. METHOD: We compared the individual depressive symptoms of 145 non-demented depressed patients with PD and 100 depressed patients without PD by comparing item scores on the Montgomery-Asberg Depression Rating Scale by way of MANCOVA. RESULTS: The severity of depression and the level of cognitive functioning in depressed PD patients were comparable with that of depressed control subjects. However, patients with PD showed significant less reported sadness, less anhedonia, less feelings of guilt and, slightly less loss of energy, but more concentration problems than depressed control subjects. CONCLUSION: The profile of depressive symptoms in PD differs from that in depressed subjects without PD. This finding is important for the conceptualisation and clinical diagnosis of depression in PD. PMID- 16477587 TI - An open-label study of escitalopram (Lexapro) for the treatment of 'Depression of Alzheimer's disease' (dAD). AB - BACKGROUND: Depression is a frequent neuropsychiatric complication of Alzheimer's Disease. METHODS: This study investigated the safety and effectiveness of escitalopram (LEXAPRO) for depression in AD (dAD) as defined by the NIMH consensus criteria in an 8-week, open-label treatment study. CONCLUSION: Escitalopram was efficacious and safe for the treatment of dAD in this study. Larger, controlled studies are warranted to further assess the efficacy for mood and behavioral disturbances in this medically fragile population. PMID- 16477588 TI - Use of antipsychotic medications among elderly residents in long-term institutional care: a three-year follow-up. AB - OBJECTIVE: To analyse the use of antipsychotic medications, change over time and associated factors in a three-year follow-up among elderly residents in long-term institutional care. DESIGN: Retrospective study was designed with three identical cross-sectional samples originating from the same long-term care facilities, and collected 1 July to 31 December in 2001, 2002 and 2003, in Finland. These were extracted from the Resident Assessment Instrument (RAI) database, based on Minimum Data Set (MDS) assessments. SETTING: Of the data providers 16 were hospital-based institutions and 25 residential homes. PARTICIPANTS: Each of the data sets included 3,662-3,867 resident assessments. RESULTS: The prevalence use of one or more antipsychotic decreased from 42% in 2001 to 39% in 2003. The overall confounder-adjusted decrease in antipsychotic use was not statistically significant. However, the use of antipsychotics decreased among residents who had wandering as a behavioural problem (OR 0.79, 95% CI 0.63-0.99) and increased among residents with concomitant use of anxiolytic medications (OR 1.23, 95% CI 1.03-1.48). CONCLUSIONS: The use of antipsychotic medications among residents in long-term institutional care was common and the caring patterns were quite stable during the observation period. Adequate indications may not have been achieved in all cases. More attention should be paid to the appropriate use of antipsychotics in this frail population. PMID- 16477589 TI - Leg length, cognitive impairment and cognitive decline in an African-Caribbean population. AB - BACKGROUND: Shorter leg length is associated with an adverse environment in early childhood and has been found to be associated with a variety of disorders occurring in mid- to late-life, including dementia in a Korean population. In a community population of African-Caribbean elders, in whom leg length had been measured, we sought to compare associations with cognitive impairment at baseline and cognitive decline over a three-year follow-up period. METHODS: Of 290 African Caribbean residents in south London recruited at baseline, 216 (74%) were re interviewed after a three-year period and 203 had sufficient data for this analysis. Cognitive impairment was derived as a binary category from a battery of cognitive tests administered at baseline and cognitive decline was derived from change in performance on a subset of these tests. Leg length (iliac crest to lateral malleolus) was also measured. RESULTS: Shorter leg length was associated with female sex, lower occupational social class and reported hypertension and diabetes. Shorter leg length (lowest quartile) was significantly associated with cognitive impairment but there were no apparent associations with cognitive decline. The association with cognitive impairment was independent of age, sex and education. Social class appeared to be an important mediating factor. CONCLUSIONS: Shorter leg length may be a marker of early life stressors which result in reduced cognitive reserve. Interestingly this association was mediated more strongly by social class (previous occupational status) than by education in this population. PMID- 16477594 TI - Positive reinforcement training affects hematologic and serum chemistry values in captive chimpanzees (Pan troglodytes). AB - Positive reinforcement training (PRT) techniques have received considerable attention for their stress reduction potential in the behavioral management of captive nonhuman primates. However, few published empirical studies have provided physiological data to support this position. To address this issue, PRT techniques were used to train chimpanzees (Pan troglodytes) to voluntarily present a leg for an intramuscular (IM) injection of anesthetic. Hematology and serum chemistry profiles were collected from healthy chimpanzees (n=128) of both sexes and various ages during their routine annual physical examinations over a 7 year period. Specific variables potentially indicative of acute stress (i.e., total white blood cell (WBC) counts, absolute segmented neutrophils (SEG), glucose (GLU) levels, and hematocrit (HCT) levels) were analyzed to determine whether the method used to administer the anesthetic (voluntary present for injection vs. involuntary injection) affected the physiological parameters. Subjects that voluntarily presented for an anesthetic injection had significantly lower mean total WBC counts, SEG, and GLU levels than subjects that were involuntarily anesthetized by more traditional means. Within-subjects analyses revealed the same pattern of results. This is one of the first data sets to objectively demonstrate that PRT for voluntary presentation of IM injections of anesthetic can significantly affect some of the physiological measures correlated with stress responses to chemical restraint in captive chimpanzees. PMID- 16477595 TI - Living in nonbreeding groups: an alternative strategy for maturing gorillas. AB - The one-male reproductive strategy implies that maturing males are temporarily excluded from reproduction. In gorillas, these excluded males live either solitarily or in nonbreeding groups (NBGs) that are devoid of adult females. The dynamics of NBGs are not well known. In this study, which was conducted on a gorilla population (Gorilla gorilla gorilla) of 377 individuals that visited the Lokoue clearing in the Republic of Congo, we detail how the NBGs formed, and analyze their dynamics according to age-sex classes, the relatedness of members, and the origin and destination of transferring individuals. We discuss the potential benefits gained by individuals living in these groups. The NBGs included mainly immature males, most of which appeared to have migrated voluntarily from their natal groups. Some individuals (including juvenile females) came from disbanded breeding groups (BGs). Migrants preferentially joined NBGs that included a silverback male. Their dispersal patterns were not determined by their degree of relatedness, but they tended to associate with related silverbacks. In this way, the migrants could enhance their protection against predators and gain experience with different environmental conditions. By tolerating and protecting offspring, aging silverbacks could enhance their inclusive fitness. Finally, young and healthy silverbacks could increase their likelihood of forming a future BG when unrelated females joined them. PMID- 16477590 TI - Are the Mallory bodies and intracellular hyaline bodies in neoplastic and non neoplastic hepatocytes related? AB - Mallory bodies (MBs) and intracellular hyaline bodies (IHBs) are cytoplasmic hepatocellular inclusions that consist of aggregated proteins. MBs are characteristically associated with alcoholic and non-alcoholic steatohepatitis, but may also be found in chronic cholestatic and metabolic (eg copper intoxication) diseases and hepatocellular neoplasms, particularly hepatocellular carcinomas. IHBs have hitherto only been described in hepatocellular carcinoma cells. In the present study hepatocellular carcinomas (HCCs) and a case of idiopathic copper toxicosis were evaluated with respect to the presence and mutual relationship of MBs and IHBs. IHBs alone were present in 8.6%, MBs alone in 16.1% and both types of inclusion in 7.5% of HCCs. It is shown that IHBs may also occur in non-neoplastic hepatocytes in association with idiopathic copper toxicosis, together with MBs. In HCCs and idiopathic copper toxicosis, MBs and IHBs may be present within the same cell. Moreover, hybrid inclusions holding an intermediate position between MBs and IHBs regarding light microscopy, ultrastructure and composition exist. MBs and IHBs contain p62, a stress inducible adapter protein, as the major constituent. In MBs p62 is associated with keratins, whereas classical IHBs lack keratins. Light microscopic, electron microscopic and immunohistochemical data suggest a close pathogenetic relationship between MBs and IHBs. Both types of inclusion are the result of over expression and accumulation of the stress protein p62. If p62 is induced alone, or at least prevails, IHBs may arise by aggregation. However, if abnormal keratins are present in addition to p62, p62 associates and co-aggregates with keratins, finally leading to classical MBs. PMID- 16477596 TI - Seasonal variation of diet and food availability in a group of Sichuan snub-nosed monkeys in Shennongjia Nature Reserve, China. AB - We studied the diet and food availability of a group of Sichuan snub-nosed monkeys for 14 months (July 2003 to September 2004, except for February) in the Shennongjia Nature Reserve, China. This species is primarily a lichen eater, with lichens (Usneaceae) accounting for 43.28% of feeding records (n=3,452). Other food types in the diet were young leaves (28.71%), fruits or seeds (14.57%), buds (5.36%), mature leaves (3.51%), herbs (2.09%), bark (1.36%), and flowers (1.13%). The monkeys used 23 plant species. Their diet showed a complicated seasonal variation: the monthly diet varied from primarily lichens in November-April, to a mixture of leaves and lichens in May-July, to a mixture of fruits or seeds and lichens in August-October (the latter depended on annual fruit and seed availability). The proportion of fruits or seeds in the diet was negatively correlated with that of lichens, which suggests that the monkeys prefer fruits or seeds to lichens when all of these items are available. The fruit or seed availability varied greatly between the two study years. The proportion of lichens, young leaves, flowers, and fruits or seeds in the diet was positively associated with their availability. The monkeys appeared to be selective feeders. They preferred 10 tree species for plant parts, and nine tree species for lichens. The selection index of tree species for lichens was positively related to lichen coverage per branch on tree species, demonstrating that the monkeys preferred tree species with abundant lichens, as well as dead trees for lichens. The results suggest that dead-tree harvesting in the reserve could significantly reduce the quality of habitat for these monkeys, and should therefore be prohibited. Connus controversa, Cerasus discadenia, Salix willichiana, and Malus halliana should be conserved as top priority species because the monkeys preferred them for both their vegetative parts and the lichens that grow on them. PMID- 16477597 TI - Distribution and density of Callimico goeldii in the Department of Pando, Bolivia. AB - A survey of the distribution and density of Callimico goeldii was conducted at five sites across northwestern Bolivia, in the Department of Pando. C. goeldii was found at two sites north of the Manuripi River, with high densities at one site located along the Acre River. Estimates of habitat availability at these sites suggest that C. goeldii reaches high densities in areas with low human density and well established and extensive bamboo forests. These results, when reviewed with those of prior studies, indicate that the patchy distribution of C. goeldii in Pando is a result of both riverine barriers and the availability of bamboo habitat. PMID- 16477598 TI - Lemur responses to edge effects in the Vohibola III classified forest, Madagascar. AB - Forest edges are dynamic zones characterized by the penetration (to varying depths and intensities) of conditions from the surrounding environment (matrix) into the forest interior. Although edge effects influence many tropical organisms, they have not been studied directly in primates. Edge effects are particularly relevant to lemurs because of the highly fragmented forest landscapes found in Madagascar. In this study, data are presented regarding how the densities of six lemur species (Avahi laniger, Cheirogaleus major, Eulemur rubriventer, Hapalemur griseus griseus, Microcebus rufus, and Propithecus diadema edwardsi) varied between six 500-m interior transects and six 500-m edge transects in the Vohibola III Classified Forest in SE Madagascar. Diurnal (n = 433) and nocturnal (n = 128) lemur surveys were conducted during June-October 2003 and May-November 2004. A. laniger, E. rubriventer, and H. g. griseus exhibited a neutral edge response (no differences in densities between habitats). M. rufus and P. d. edwardsi had a positive edge response (higher densities in edge habitats), which may be related to edge-related variations in food abundance and quality. Positive edge responses by M. rufus and P. d. edwardsi may ultimately be detrimental due to edge-related anthropogenic factors (e.g., hunting by local people). The negative edge response exhibited by C. major (lower densities in edge habitats) may result from heightened ambient temperatures that inhibit torpor in edge habitats. PMID- 16477599 TI - Amino acid sequence predicts folding rate for middle-size two-state proteins. AB - The significant correlation between protein folding rates and the sequence predicted secondary structure suggests that folding rates are largely determined by the amino acid sequence. Here, we present a method for predicting the folding rates of proteins from sequences using the intrinsic properties of amino acids, which does not require any information on secondary structure prediction and structural topology. The contribution of residue to the folding rate is expressed by the residue's Omega value. For a given residue, its Omega depends on the amino acid properties (amino acid rigidity and dislike of amino acid for secondary structures). Our investigation achieves 82% correlation with folding rates determined experimentally for simple, two-state proteins studied until the present, suggesting that the amino acid sequence of a protein is an important determinant of the protein-folding rate and mechanism. PMID- 16477600 TI - Comparison of different enzyme-immunoassays for assessment of adrenocortical activity in primates based on fecal analysis. AB - Most studies published to date that used fecal glucocorticoid measurements to assess adrenocortical activity in primate (and many nonprimate) species applied a specific cortisol or corticosterone assay. However, since these native glucocorticoids are virtually absent in the feces of most vertebrates, including primates, the validity of this approach has recently been questioned. Therefore, the overall aim of the present study was to assess the validity of four enzyme immunoassays (EIAs) using antibodies raised against cortisol, corticosterone, and reduced cortisol metabolites (two group-specific antibodies) for assessing adrenocortical activity using fecal glucocorticoid metabolite (GCM) measurements in selected primate species (marmoset, long-tailed macaque, Barbary macaque, chimpanzee, and gorilla). Using physiological stimulation of the hypothalamo pituitary-adrenocortical (HPA) axis by administering exogenous ACTH or anesthesia, we demonstrated that at least two assays detected the predicted increase in fecal GCM levels in response to treatment in each species. However, the magnitude of response varied between assays and species, and no one assay was applicable to all species. While the corticosterone assay generally was of only limited suitability for assessing glucocorticoid output, the specific cortisol assay was valuable for those species that (according to high-performance liquid chromatography (HPLC) analysis data) excreted clearly detectable amounts of authentic cortisol into the feces. In contrast, in species in which cortisol was virtually absent in the feces, group-specific assays provided a much stronger signal, and these assays also performed well in the other primate species tested (except the marmoset). Collectively, the data suggest that the reliability of a given fecal glucocorticoid assay in reflecting activity of the HPA axis in primates clearly depends on the species in question. Although to date there is no single assay system that can be used successfully across species, our data suggest that group-specific assays have a high potential for cross-species application. Nevertheless, regardless of which GC antibody is chosen, our study clearly reinforces the necessity of appropriately validating the respective assay system before it is used. PMID- 16477601 TI - Nearest-neighbor effects and structural preferences in dipeptides are a function of the electronic properties of amino acid side-chains. AB - The electronic properties of amino acid side-chains are emerging as an important factor in the preference for secondary structure in proteins. These properties have not been fully characterized, nor has their role in the behavior of peptides been explored in any detail. The present studies sought to evaluate several possibilities: 1) that hydrophilicity can be expressed solely in electronic terms, 2) that substituent effects of side-chains extend across the peptide bond, and (3) nearest-neighbor effects in dipeptides correlate with secondary structural preferences. Quantum mechanics (QM) calculations were used to define the electronic properties of individual amino acids and dipeptides. It was found that the hydrophilicity of an amino acid side-chain can be accurately represented as a function of the electron densities of its component atoms. In addition, the nature of an amino acid in the second position of a dipeptide affects the electronic properties (Mulliken populations and electron densities) of the main chain atoms of the first residue. Certain electronic features of the dipeptides strongly correlated with propensity for secondary structure. Specifically, Mulliken population data at the Calpha atom and N atom predicted preference for alpha-helices versus coil and strand conformations, respectively. Analysis of dipeptides arrayed in either helical or extended structures revealed lengthening of main-chain bonds in the alpha-helical conformations. A thorough characterization of the electronic properties of amino acids and short peptide segments may provide a better understanding of the forces that determine secondary structure in proteins. PMID- 16477602 TI - Crystal structures of two putative phosphoheptose isomerases. PMID- 16477603 TI - Convergent evolution of sexual shape dimorphism in Diptera. AB - Several patterns of sexual shape dimorphism, such as male body elongation, eye stalks, or extensions of the exoskeleton, have evolved repeatedly in the true flies (Diptera). Although these dimorphisms may have evolved in response to sexual selection on male body shape, conserved genetic factors may have contributed to this convergent evolution, resulting in stronger phenotypic convergence than might be expected from functional requirements alone. I compared phenotypic variation in body shape in two distantly related species exhibiting sexually dimorphic body elongation: Prochyliza xanthostoma (Piophilidae) and Telostylinus angusticollis (Neriidae). Although sexual selection appears to act differently on male body shape in these species, they exhibited strikingly similar patterns of sexual dimorphism. Likewise, patterns of within-sex shape variation were similar in the two species, particularly in males: relative elongation of the male head capsule, antenna, and legs was associated with reduced head capsule width and wing length, but was nearly independent of variation in thorax length. However, the two species presented contrasting patterns of static allometry: male sexual traits exhibited elevated allometric slopes in T. angusticollis, but not in P. xanthostoma. These results suggest that a shared pattern of covariation among traits may have channeled the evolution of sexually dimorphic body elongation in these species. Nonetheless, static allometries may have been shaped by species-specific selection pressures or genetic architectures. PMID- 16477604 TI - Comparison of wing morphology in three birds of prey: correlations with differences in flight behavior. AB - Flight is the overriding characteristic of birds that has influenced most of their morphological, physiological, and behavioral features. Flight adaptations are essential for survival in the wide variety of environments that birds occupy. Therefore, locomotor structure, including skeletal and muscular characteristics, is adapted to reflect the flight style necessitated by different ecological niches. Red-tailed hawks (Buteo jamaicensis) soar to locate their prey, Cooper's hawks (Accipiter cooperii) actively chase down avian prey, and ospreys (Pandion haliaetus) soar and hover to locate fish. In this study, wing ratios, proportions of skeletal elements, and relative sizes of selected flight muscles were compared among these species. Oxidative and glycolytic enzyme activities of several muscles were also analyzed via assays for citrate synthase (CS) and for lactate dehydrogenase (LDH). It was found that structural characteristics of these three raptors differ in ways consistent with prevailing aerodynamic models. The similarity of enzymatic activities among different muscles of the three species shows low physiological differentiation and suggests that wing architecture may play a greater role in determining flight styles for these birds. PMID- 16477605 TI - Cystic metastasis from head and neck squamous cell cancer: a distinct disease variant? AB - BACKGROUND: Head and neck squamous cell carcinoma (HNSCC) commonly spreads to regional deep cervical nodes. In most cases, these metastases present as firm, solid masses in the designated lymph node chains. A distinct subset of metastatic nodes present as cystic masses, with most of the volume made up of a liquid center surrounded by a thin solid rim. It has been observed that certain squamous cell carcinoma (SCC) subsites are more likely to produce metastases that are cystic. These sites predominantly include primary tumors of tonsil tissue from Waldeyer's ring. In the past, these cystic cancers often have been erroneously diagnosed as branchiogenic carcinomas, that is, a branchial cleft cyst that has undergone malignant degeneration. Today, most authors have concluded that so called branchiogenic carcinomas are actually cystic metastases in the neck probably arising from an oropharyngeal primary SCC. The purpose of this work is to consider the phenomenon of cystic lymph node metastasis in head and neck cancer in depth. METHODS: A review of the relevant English-language literature linking cystic metastasis and head and neck cancer was performed. RESULTS: These studies indicate that lateral cystic masses in adults often represent an occult primary cancer originating in the epithelium within Waldeyer's ring. CONCLUSIONS: Adult patients who are initially seen with a lateral cystic neck mass must be presumed to have a cancer until proven otherwise. The mass should be biopsied by fine-needle aspiration (FNA). However, negative FNA findings may be misleading; therefore, an excisional biopsy and examination under anesthesia with directed biopsies of Waldeyer's ring and bilateral tonsillectomy should be considered a part of the diagnostic workup. PMID- 16477606 TI - Clinical curiosity: cribriform-morular variant of papillary thyroid carcinoma. AB - BACKGROUND: There is an increasing awareness of the association of papillary thyroid carcinoma and familial adenomatous polyposis (FAP). Although the incidence is rare, most tend to occur in women. Several authors have described a distinctive histologic variant of papillary thyroid carcinoma, the cribriform morular variant, which is associated with FAP but also may be encountered in patients with non-FAP. This diagnosis may precede the symptoms of colorectal polyposis. METHODS: A healthy 36-year-old woman was seen with a left thyroid nodule, and a 34-year-old woman with FAP was seen with a right thyroid nodule; both masses were suspicious for papillary thyroid carcinoma. Both patients underwent total thyroidectomy. RESULTS: Pathologic examination of both specimens revealed papillary thyroid carcinoma, cribriform-morular variant. The first patient subsequently underwent colonoscopy, which was negative for polyposis. CONCLUSIONS: Patients diagnosed with the cribriform-morular variant of papillary thyroid cancer should be screened for the presence of FAP. PMID- 16477607 TI - Black thyroid resulting from short-term doxycycline use: case report, review of the literature, and discussion of implications. AB - BACKGROUND: Black thyroid pigmentation has been considered pathognomonic for chronic minocycline ingestion for more than 30 years. Although never conclusively linked to overt disease, evidence clearly exists that minocycline is a competitive inhibitor of thyroid peroxidase in metabolically active thyroid tissue. This offers a potential mechanism of pigment accumulation, which can account for the occasional finding of hypopigmentation in thyroid carcinomas. To our knowledge, an association with tetracycline derivatives other than minocycline has not been documented. METHODS: Herein is a case report of a patient with gross black thyroid tissue containing a hypopigmented papillary thyroid carcinoma. Twelve days before surgery, the patient was placed on doxycycline, a tetracycline derivative, to optimize an in vitro fertilization regimen. RESULTS: The gross specimen was diffusely black in color with a 1.5-cm hypopigmented focus of papillary thyroid cancer. Hematoxylin-eosin staining, as well as electron micrographs, was consistent with findings associated with minocycline ingestion. CONCLUSIONS: Black thyroid is rare and only previously associated with chronic minocycline ingestion. This report documents a black thyroid in a patient after short-term exposure to doxycycline. Pigment accumulation in normal tissue is thought to occur by inhibition of thyroid peroxidase. Minocycline is a competitive inhibitor of this enzyme in its native configuration. Thyroid carcinomas are known to have abnormal thyroid peroxidase, which could account for reports of hypopigmented tumors within grossly darkened thyroid tissue. Hypopigmented foci within such "black thyroid" deserve through pathologic examination. PMID- 16477608 TI - Cognitive-behavioral profiles of females with the fragile X mutation. AB - The fragile X (FRAXA) mutation is typically manifested as either a full mutation (FM) or premutation (PM), and is often associated with some form of learning impairment. The study by Lachiewicz et al. in this issue suggests that females with the FM or PM exhibit a specific profile of strengths in verbal abilities and significant weaknesses in quantitative skills. We examined 17 females with either the FM or PM using a standard cognitive-behavioral battery consisting of the Stanford-Binet (4th Edition; SBFE) and the Vineland Adaptive Behavior Scale (VABS). Although we found the expected differences in composite IQ scores and adaptive behavior composite scores (DQ) between FM and PM females, we found no significant differences between verbal and quantitative reasoning in either group. PMID- 16477609 TI - Noradrenergic regulation of galanin expression in the supraoptic nucleus in the rat hypothalamus. An ex vivo study. AB - Galanin is coexpressed with vasopressin and oxytocin in magnocellular neurons of the rat neuroendocrine hypothalamus. Various physiological stimuli, such as osmotic stimulation or lactation, that affect vasopressin and oxytocin expression and release also modulate galanin expression. Magnocellular neurons are highly innervated by noradrenergic inputs from the brainstem. The noradrenergic system plays a critical excitatory role in the activation of vasopressin-expressing and oxytocin-expressing neurons. Here, we have evaluated the possible regulation of Gal expression by noradrenaline in the magnocellular neurons of supraoptic nucleus in an ex vivo acute model of rat hypothalamic slices. The slices containing the supraoptic nucleus were incubated with 10(-4) M noradrenaline for 1 or 4 hr. The levels of galanin and galanin mRNA were estimated by semiquantitative immunohistochemistry and in situ hybridization, respectively. Our results show that the amount of galanin-immunopositive material in the cell bodies of the magnocellular neurons increased significantly after incubation with noradrenaline compared with control slices at the same time point and that this effect was more pronounced after 4 hr than after 1 hr. In situ hybridization showed that radiolabeling of the supraoptic nucleus with a radioactive galanin probe increased slightly after 1 hr of incubation and increased considerably after 4 hr of incubation with noradrenaline. Our study shows that galanin may be a target in the regulation of the hypothalamic magnocellular-neurohypophysial system by noradrenaline. PMID- 16477610 TI - Semisynthetic sphingoglycolipid LIGA20 is neuroprotective against human immunodeficiency virus-gp120-mediated apoptosis. AB - Apoptosis and neuronal atrophy are commonly seen in patients infected with the human immunodeficiency virus type 1 (HIV-1) in the late phase of infection. The HIV-1 envelope glycoprotein gp120 has been suggested to be a causal agent of neuronal loss. Therefore, blocking gp120 neurotoxicity may be an effective way to reduce the neuronal degeneration seen in HIV patients. Brain-derived neurotrophic factor (BDNF) prevents gp120-mediated apoptosis in cerebellar granule cells. However, BDNF poorly crosses the blood-brain barrier and therefore may not be a suitable therapy for HIV patients. LIGA20 is a semisynthetic sphingoglycolipid that may be a valid alternative to BDNF. In fact, it has been shown that LIGA20 mimics the neuroprotective properties of BDNF. The present study was undertaken to characterize the relative potency of LIGA20 to antagonize gp120-mediated apoptosis. Cerebellar granule cells were exposed to gp120IIIB (5 nM) or stromal cell derived factor-1 (SDF), the natural ligand for the CXCR4 receptor to which gp120 binds, alone or in combination with LIGA20 (5 microM), and cell death/survival was determined 12 and 24 hr later by various markers of apoptosis. LIGA20 blocked the neurotoxic effect of gp120 and SDF. The neurotrophic effect of LIGA20 was reversed by K252a, a tyrosine kinase inhibitor used to block TrkB signaling, suggesting the involvement of TrkB activation. These findings provide the rationale for exploring the ability of compounds that mimic BDNF activity to reduce neuronal cell death in HIV-1-positive patients. PMID- 16477611 TI - Effect of maturation stage at cryopreservation on post-thaw cytoskeleton quality and fertilizability of equine oocytes. AB - Oocyte cryopreservation is a potentially valuable technique for salvaging the germ-line when a valuable mare dies, but facilities for in vitro embryo production or oocyte transfer are not immediately available. This study examined the influence of maturation stage and freezing technique on the cryopreservability of equine oocytes. Cumulus oocyte complexes were frozen at the immature stage (GV) or after maturation in vitro for 30 hr (MII), using either conventional slow freezing (CF) or open pulled straw vitrification (OPS); cryoprotectant-exposed and untreated nonfrozen oocytes served as controls. After thawing, GV oocytes were matured in vitro, and MII oocytes were incubated for 0 or 6 hr, before staining to examine meiotic spindle quality by confocal microscopy. To assess fertilizability, CF MII oocytes were subjected to intracytoplasmic sperm injection (ICSI) and cultured in vitro. At 12, 24, and 48 hr after ICSI, injected oocytes were fixed to examine their progression through fertilization. Both maturation stage and freezing technique affected oocyte survival. The meiosis resumption rate was higher for OPS than CF for GV oocytes (28% vs. 1.2%; P < 0.05), but still much lower than for controls (66%). Cryopreserving oocytes at either stage induced meiotic spindle disruption (37% 67% normal spindles vs. 99% in controls; P < 0.05). Among frozen oocytes, however, spindle quality was best for oocytes frozen by CF at the MII stage and incubated for 6 hr post-thaw (67% normal); since this combination of cryopreservation/IVM yielded the highest proportion of oocytes reaching MII with a normal spindle (35% compared to <20% for other groups), it was used when examining the effects of cryopreservation on fertilizability. In this respect, the rate of normal fertilization for CF MII oocytes after ICSI was much lower than for controls (total oocyte activation rate, 26% vs. 56%; cleavage rate at 48 hr, 8% vs. 42%: P < 0.05). Thus, although IVM followed by CF yields a respectable percentage of normal-looking MII oocytes (35%), their ability to support fertilization is severely compromised. PMID- 16477612 TI - Leukemia inhibitory factor is produced by myelin-reactive T cells from multiple sclerosis patients and protects against tumor necrosis factor-alpha-induced oligodendrocyte apoptosis. AB - In multiple sclerosis (MS), damage to oligodendrocytes is believed to be caused by an aberrant immune response initiated by autoreactive T cells. Increasing evidence indicates that these T cells are not exclusively detrimental but might also exert protective effects. We report for the first time that myelin-reactive T-cell clones from eight MS patients (6/19) and five healthy controls (4/11) produce leukemia inhibitory factor (LIF), a member of the neuropoietic family of neurotrophins. In addition, T-cell clones specific for tetanus toxoid, CD4(+) and CD8(+) T cells, and monocytes, but not B cells, secreted LIF. LIF-producing T lymphocytes and macrophages were also identified immunohistochemically in both active and chronic-active MS lesions. We further demonstrated dose-dependent protective effects of LIF on tumor necrosis factor-alpha-induced apoptosis of oligodendrocytes. In conclusion, our data demonstrate that peripheral and CNS infiltrating T cells from MS patients produce LIF, a protective factor for oligodendrocytes. This study emphasizes that secretion of LIF may contribute to the neuroprotective effects of autoreactive T cells. PMID- 16477613 TI - Femtosecond laser ablation elemental mass spectrometry. AB - Laser ablation mass spectrometry (LA-MS) has always been an interesting method for the elemental analysis of solid samples. Chemical analysis with a laser requires small amounts of material. Depending on the analytical detection system, subpicogram quantities may be sufficient. In addition, a focused laser beam permits the spatial characterization of heterogeneity in solid samples typically with micrometer resolution in terms of lateral and depth dimensions. With the advent of high-energy, ultra-short pulse lasers, new possibilities arise. The task of this review is to discuss the principle differences between the ablation process of short (>1 ps) and ultra-short (<1 ps) pulses. Based on the timescales and the energy balance of the process that underlies an ablation event, it will be shown that ultra-short pulses are less thermal and cause less collateral damages than longer pulses. The confinement of the pulse energy to the focal region guarantees a better spatial resolution in all dimensions and improves the analytical figures of merit (e.g., fractionation). Applications that demonstrate these features and that will be presented are in-depth profiling of multi-layer samples and the elemental analysis of biological materials. PMID- 16477614 TI - Trkb receptors modulation of glutamate release is limited to a subset of nerve terminals in the adult rat hippocampus. AB - Brain-derived neurotrophic factor (BDNF) modulates glutamatergic excitatory transmission in hippocampal primary cultures by acting at a presynaptic locus. Although it has been suggested that BDNF also modulates adult hippocampus glutamatergic transmission, this remains a matter of controversy. To clarify a putative role for this neurotrophin in the modulation of glutamate release we applied exogenous BDNF to isolated adult rat hippocampal nerve terminals. BDNF, at 100 ng/ml, potentiated by 25% the K(+)-evoked release of [(3)H]glutamate from hippocampal synaptosomes. The small effect of BDNF on [(3)H]glutamate release correlated with a modest increase in phospholipase Cgamma (PLCgamma) phosphorylation, and with the lack of effect of BDNF on extracellular-signal regulated kinase (ERK) and Akt phosphorylation. Immunocytochemistry studies demonstrated that only about one-third of glutamatergic synaptosomes were positive for TrkB immunoreactivity. Furthermore, biotinylation and subsynaptic fractionation studies showed that only one-fourth of total full-length TrkB was present at the plasma membrane, evenly distributed between the presynaptic active zone and the postsynaptic density. These results indicate that BDNF modulates synaptic transmission presynaptically in a small subset of hippocampal glutamatergic synapses that contain TrkB and that express the receptor on the plasma membrane. PMID- 16477615 TI - Neuropathies associated with excessive exposure to lead. AB - Exposure to lead is a ubiquitous problem of the modern era. The majority of cases of all forms of lead intoxication, especially lead neuropathy, result from industrial exposure. In the Western world meticulous monitoring in industry has reduced the risk of overt lead neuropathy. The classic form of lead neuropathy consists of weakness that primarily involves the wrist and finger extensors but which later spreads to other muscles. There is only minimal sensory involvement. Less commonly, there is a more typical toxic neuropathy with distally accentuated sensory and motor involvement. The motor neuropathy is more likely to develop following relatively short-term exposure to high lead concentrations and evolves in a subacute fashion. Prognosis for recovery is good as long as exposure is terminated promptly. The distal sensory and motor neuropathy develops after many years of exposure, evolves more slowly, and recovery is less certain. There is a generally weak relationship between the development of lead neuropathy and blood lead levels, at least for the subacute motor neuropathy, leading to speculation that the metabolic basis for the neuropathy is interference with porphyrin metabolism. Lead intoxication in humans causes axonal degeneration, but in some other species it causes a primarily demyelinating neuropathy. It should be possible to prevent lead neuropathy by good industrial hygiene. Close monitoring should identify excessive lead exposure before it causes overt neuropathy. If evidence of excessive exposure is found or if overt neuropathy develops, exposure must be terminated immediately. The role of chelation therapy in the treatment of lead neuropathy is controversial. PMID- 16477616 TI - S100B protects LAN-5 neuroblastoma cells against Abeta amyloid-induced neurotoxicity via RAGE engagement at low doses but increases Abeta amyloid neurotoxicity at high doses. AB - At the concentrations normally found in the brain extracellular space the glial derived protein, S100B, protects neurons against neurotoxic agents by interacting with the receptor for advanced glycation end products (RAGE). It is known that at relatively high concentrations S100B is neurotoxic causing neuronal death via excessive stimulation of RAGE. S100B is detected within senile plaques in Alzheimer's disease, where its role is unknown. The present study was undertaken to evaluate a putative neuroprotective role of S100B against Abeta amyloid induced neurotoxicity. We treated LAN-5 neuroblastoma cultures with toxic amounts of Abeta25-35 amyloid peptide. Our results show that at nanomolar concentrations S100B protects cells against Abeta-mediated cytotoxicity, as assessed by 3-(4,5 dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and terminal deoxynucleotidyl transferase-mediated dUTP-fluorescein isothiocyanate nick end labeling (TUNEL) experiments, by countering the Abeta-mediated decrease in the expression of the anti-apoptotic factor Bcl-2. This effect depends on S100B binding to RAGE because S100B is unable to contrast Abeta-mediated neurotoxicity in neurons overexpressing a signaling-deficient RAGE mutant lacking the cytosolic and transducing domain. Our data suggest that at nanomolar doses S100B counteracts Abeta peptide neurotoxicity in a RAGE-mediated manner. However, at micromolar doses S100B is toxic to LAN-5 cells and its toxicity adds to that of the Abeta peptide, suggesting that additional molecular mechanisms may be involved in the neurotoxic process. PMID- 16477617 TI - Sarcoplasmic reticulum: the dynamic calcium governor of muscle. AB - The sarcoplasmic reticulum (SR) provides feedback control required to balance the processes of calcium storage, release, and reuptake in skeletal muscle. This balance is achieved through the concerted action of three major classes of SR calcium-regulatory proteins: (1) luminal calcium-binding proteins (calsequestrin, histidine-rich calcium-binding protein, junctate, and sarcalumenin) for calcium storage; (2) SR calcium release channels (type 1 ryanodine receptor or RyR1 and IP3 receptors) for calcium release; and (3) sarco(endo)plasmic reticulum Ca2+ ATPase (SERCA) pumps for calcium reuptake. Proper calcium storage, release, and reuptake are essential for normal skeletal muscle function. We review SR structure and function during normal skeletal muscle activity, the proteins that orchestrate calcium storage, release, and reuptake, and how phenotypically distinct muscle diseases (e.g., malignant hyperthermia, central core disease, and Brody disease) can result from subtle alterations in the activity of several key components of the SR calcium-regulatory machinery. PMID- 16477618 TI - Pro-apoptotic role of c-Jun in NMDA-induced neurotoxicity in the rat retina. AB - We examined the role of c-Jun on N-methyl-D-aspartate (NMDA)-induced neurotoxicity in the rat retina. An increase in c-Jun mRNA, c-Jun protein and phosphorylated c-Jun (p-c-Jun) levels in the retina was detected 3 hr after intravitreal injection of NMDA (20 nmol). These levels peaked after 12 hr, and then returned to their control levels by 24 hr. c-Jun and p-c-Jun immunoreactivities were observed in the retinal ganglion cell layer (RGCL), especially in retinal ganglion cells (RGCs), and in the inner nuclear layer (INL) 12 hr after NMDA injection, and terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL)-positive cells were immunopositive for c-Jun and p-c Jun. A c-Jun antisense oligodeoxynucleotide (AS ODN), which was simultaneously injected with NMDA, penetrated the cells in the RGCL and the INL, suppressed the NMDA-induced increase in c-Jun and p-c-Jun protein levels and reduced the number of TUNEL-positive cells in the RGCL 12 hr after the injection. The protective effect of c-Jun AS ODN on the NMDA-treated retina was also shown by the RGCL cell count and measurement of the IPL thickness, as well as by quantitative real-time PCR analysis of Thy-1 mRNA 7 days after the injection. These results suggest that c-Jun synthesis and phosphorylation participate in NMDA-induced neuronal cell death. PMID- 16477619 TI - Early social enrichment augments adult hippocampal BDNF levels and survival of BrdU-positive cells while increasing anxiety- and "depression"-like behavior. AB - Early experiences affect brain function and behavior at adulthood. Being reared in a communal nest (CN), consisting of a single nest where three mothers keep their pups together and share care-giving behavior from birth to weaning (postnatal day [PND] 25), provides an highly socially stimulating environment to the developing pup. Communal nest characterizes the natural ecologic niche of many rodent species including the mouse. At adulthood, CN reared mice, compared to mice reared in standard nesting laboratory condition (SN), show an increase in BDNF protein levels and longer survival of BrdU-positive cells in the hippocampus. Open field and elevated plus maze results indicate that CN mice, although showing levels of exploratory and locomotor activity similar to those of SN mice, displayed increased anxiety-like behavior, performing more thigmotaxis in the open field and spending less time in the open arms of the plus maze. Furthermore, CN mice displayed higher levels of immobility behavior in the forced swim test. Overall, these findings show that CN, an highly stimulating early social environment, increases adult neuronal plasticity, as suggested by high BDNF levels and augmented number of newly generated cells in the hippocampus, which is associated to an increased anxiety- and "depression"-like behavior. These findings are discussed in the framework of the neurotrophin hypothesis of depression. PMID- 16477620 TI - Intranuclear nemaline rod myopathy. AB - The clinical, pathologic, and genetic findings of a boy with intranuclear nemaline rod myopathy are described. Serial muscle biopsies revealed myocyte nuclei containing inclusions that were immunoreactive for alpha-actinin and increased with age. Genetic analysis revealed a Val163Leu ACTA1 mutation previously associated with nemaline rod myopathy. Although initially delayed, he has reached all milestones and remains stable. These findings suggest intranuclear rods may increase with time and do not necessarily imply a poor prognosis. PMID- 16477622 TI - On the nature of cavities on protein surfaces: application to the identification of drug-binding sites. AB - In this article we introduce a new method for the identification and the accurate characterization of protein surface cavities. The method is encoded in the program SCREEN (Surface Cavity REcognition and EvaluatioN). As a first test of the utility of our approach we used SCREEN to locate and analyze the surface cavities of a nonredundant set of 99 proteins cocrystallized with drugs. We find that this set of proteins has on average about 14 distinct cavities per protein. In all cases, a drug is bound at one (and sometimes more than one) of these cavities. Using cavity size alone as a criterion for predicting drug-binding sites yields a high balanced error rate of 15.7%, with only 71.7% coverage. Here we characterize each surface cavity by computing a comprehensive set of 408 physicochemical, structural, and geometric attributes. By applying modern machine learning techniques (Random Forests) we were able to develop a classifier that can identify drug-binding cavities with a balanced error rate of 7.2% and coverage of 88.9%. Only 18 of the 408 cavity attributes had a statistically significant role in the prediction. Of these 18 important attributes, almost all involved size and shape rather than physicochemical properties of the surface cavity. The implications of these results are discussed. A SCREEN Web server is available at http://interface.bioc.columbia.edu/screen. PMID- 16477621 TI - Expression of functional CB1 cannabinoid receptors in retinoic acid differentiated P19 embryonal carcinoma cells. AB - Although primary neuronal cell cultures, usually obtained from embryonic or early postnatal rodents, have been used in vitro to study the neural cannabinoid signalling system, development of cell lines with neural properties exhibiting native expression of cannabinoid receptors is desirable. This study was undertaken to investigate the expression of CB1 and CB2 cannabinoid receptors in neurons that develop from retinoic acid (RA)-primed mouse P19 embryonal carcinoma cells. Both undifferentiated P19 cells and RA-treated P19 neurons were positive, by using reverse transcription-polymerase chain reaction (RT-PCR), for CB1 (but not CB2) mRNA. Neuronal differentiation increased the CB1 mRNA expression, and Western blotting with a CB1 receptor antibody showed a strong immunoreactive band at approximately 62 kDa in membranes from P19-derived neurons. The cannabinoid receptor agonists CP 55,940 and HU-210 produced concentration-dependent inhibition of forskolin-induced (3 microM) cyclic AMP production in the P19 derived neurons (29% at 1 microM CP 55,940 and 34% at 1 microM HU-210), which could be blocked by the CB1-selective receptor antagonist AM251, but not by the CB2-selective antagonist AM630. Furthermore, glutamate (100 microM) induced a sustained increase in [Ca2+]i in P19-derived neurons that could be concentration dependently blocked by the cannabinoid receptor agonists WIN 55,212-2. Thus, the protocol used provides an in vitro model system expressing CB1 cannabinoid receptors at the level of mRNA, protein, and AM251-sensitive agonist-induced inhibition of intracellular cyclic AMP accumulation, which may be useful to investigate the developmental regulation, expression and function of neuronal cannabinoid receptors. PMID- 16477624 TI - Empirical potential function for simplified protein models: combining contact and local sequence-structure descriptors. AB - An effective potential function is critical for protein structure prediction and folding simulation. Simplified protein models such as those requiring only Calpha or backbone atoms are attractive because they enable efficient search of the conformational space. We show residue-specific reduced discrete-state models can represent the backbone conformations of proteins with small RMSD values. However, no potential functions exist that are designed for such simplified protein models. In this study, we develop optimal potential functions by combining contact interaction descriptors and local sequence-structure descriptors. The form of the potential function is a weighted linear sum of all descriptors, and the optimal weight coefficients are obtained through optimization using both native and decoy structures. The performance of the potential function in a test of discriminating native protein structures from decoys is evaluated using several benchmark decoy sets. Our potential function requiring only backbone atoms or Calpha atoms have comparable or better performance than several residue based potential functions that require additional coordinates of side-chain centers or coordinates of all side-chain atoms. By reducing the residue alphabets down to size 10 for contact descriptors, the performance of the potential function can be further improved. Our results also suggest that local sequence structure correlation may play important role in reducing the entropic cost of protein folding. PMID- 16477623 TI - Chronically injured corticospinal axons do not cross large spinal lesion gaps after a multifactorial transplantation strategy using olfactory ensheathing cell/olfactory nerve fibroblast-biomatrix bridges. AB - Transplantation of mixed cultures containing olfactory ensheathing cell (OEC) and olfactory nerve fibroblasts (ONF) has been shown to stimulate regrowth of both acutely and chronically injured corticospinal (CS) axons across small spinal cord lesion gaps. Here, we used a multifactorial transplantation strategy to stimulate regrowth of chronically injured CS axons across large spinal cord lesion gaps. This strategy combined the transplantation of aligned OEC/ONF-biomatrix complexes, as described previously (Deumens et al. [2004] Neuroscience 125:591 604), within the lesion gap with additional OEC/ONF injections rostral and caudal to the lesion site. We show an enhanced presence of injured CS axons directly rostral to the lesion gap, with no effects on injured CS axons at or caudal to the lesion gap. Furthermore, injured CS axons did not penetrate the OEC/ONF biomatrix complex within the lesion gap. The enhanced presence of CS axons rostral to the lesion gap was not accompanied by any recovery of behavioral parameters assessed with the BBB locomotor rating scale or CatWalk gait analysis. We conclude that our multifactorial transplantation strategy should be optimized to create an OEC/ONF continuum in the injured spinal cord and thereby stimulate regrowth of injured CS axons across large spinal lesion gaps. PMID- 16477625 TI - Expression of Aurora kinases in human thyroid carcinoma cell lines and tissues. AB - The Aurora kinases are involved in the regulation of cell cycle progression, and alterations in their expression have been shown to associate with cell malignant transformation. In the present study, we demonstrated that human thyrocytes express all 3 Aurora kinases (A, B and C) at both protein and mRNA level and this expression is cell cycle-regulated. An increase in the protein level of the 3 kinases was found, with respect to normal human thyrocytes (HTU5), in the human cell lines derived from follicular (FTC-133), papillary (B-CPAP) and anaplastic (8305C) thyroid carcinomas, but not in cells derived from a follicular adenoma (HTU42). These observations were mirrored in RT-PCR experiments for Aurora-A and B. In contrast, Aurora-C mRNA levels were not significantly different among the different cell types analyzed, suggesting that posttranscriptional mechanism(s) modulate its expression. The expression at the protein level of all 3 Aurora kinases was significantly higher in 3 thyroid papillary carcinomas with respect to normal matched tissues obtained from the same patients. Similar modifications, at the mRNA level, could be observed in 7 papillary carcinoma tissues for Aurora A and B, but not for Aurora-C. In conclusion, we demonstrated that normal human thyrocytes express all 3 members of the Aurora kinase family, and their expression is amplified in malignant thyroid cell lines and tissues. These results suggest that the Aurora kinases may play a relevant role in malignant thyroid cancers, and may represent a putative therapeutic target for thyroid neoplasms. PMID- 16477626 TI - Enhanced antiangiogenic therapy with antibody-collagen XVIII NC1 domain fusion proteins engineered to exploit matrix remodeling events. AB - Antiangiogenic therapy is nowadays one of the most active fields in cancer research. The first strategies, aimed at inhibiting tumor vascularization, included upregulation of endogenous inhibitors and blocking of the signals delivered by angiogenic factors. But interaction between endothelial cells and their surrounding extracellular matrix also plays a critical role in the modulation of the angiogenic process. This study introduces a new concept to enhance the efficacy of antibody-based antiangiogenic cancer therapy strategies, taking advantage of a key molecular event occurring in the tumor context: the proteolysis of collagen XVIII, which releases the endogenous angiogenesis inhibitor endostatin. By fusing the collagen XVIII NC1 domain to an antiangiogenic single-chain antibody, a multispecific agent was generated, which was efficiently processed by tumor-associated proteinases to produce monomeric endostatin and fully functional trimeric antibody fragments. It was demonstrated that the combined production in the tumor area of complementary antiangiogenic agents from a single molecular entity secreted by gene-modified cells resulted in enhanced antitumor effects. These results indicate that tailoring recombinant antibodies with extracellular matrix-derived scaffolds is an effective approach to convert tumor progression associated processes into molecular clues for improving antibody-based therapies. PMID- 16477628 TI - Antiangiogenic combination tumor therapy blocking alpha(v)-integrins and VEGF receptor-2 increases therapeutic effects in vivo. AB - Anti-angiogenesis is a promising strategy for cancer therapy currently evaluated in clinical trials. The aim of the study was to investigate the effects of an antiangiogenic combination therapy inhibiting alpha(v)-integrins by a c(yclic)RGD peptide (EMD270179) and blocking VEGFR-2 by SU5416 on tumor angiogenesis and progression in vivo. Experiments were performed in dorsal skinfold chamber preparations of Syrian golden hamsters (60 +/- 5 g) bearing A-Mel-3 tumors. From day 3-10 after tumor-cell implantation, animals (n = 6 per group) were treated by monotherapies using the cRGD-peptide (114 mg/kg/day; i.p.), the VEGFR-2 antagonist (6 mg/kg/day; i.p.) or by the combination of both monotherapies. A control group received only the solvent DMSO. Using intravital microscopy parameters of intratumoral microcirculation were analyzed on day 5, 7 and 10. In separate experiments subcutaneous tumor growth and metastasis formation was evaluated starting therapy on day 0. Functional vessel density was significantly reduced by the combination therapy compared to that by all other groups on day 10. Although intratumoral red blood cell velocity and vessel diameters were less affected, blood flow in vessel segments and the microcirculatory perfusion index were lower after combined therapy compared to controls. In addition, we observed a significantly stronger inhibition of subcutaneous tumor growth and metastasis formation using the combination therapy. These data clearly support the concept of antiangiogenic combination therapy and demonstrate that it may especially be effective when scheduled as an early or prophylactic treatment regimen, thus avoiding angiogenesis-dependent tumor and metastasis initiation or tumor recurrence. PMID- 16477627 TI - Systemic Bcl-2 antisense oligodeoxynucleotide in combination with cisplatin cures EBV+ nasopharyngeal carcinoma xenografts in SCID mice. AB - Nasopharyngeal carcinoma (NPC) is causally linked to Epstein-Barr virus (EBV), and the EBV oncoprotein, latent membrane protein 1 (LMP-1), is expressed in the majority of NPCs. LMP-1 upregulates antiapoptotic genes, including bcl-2, and Bcl 2 protein is overexpressed in NPC. Given the antiapoptotic and chemoprotective effects of Bcl-2, it represents a rational therapeutic target in NPC. We have investigated the antitumor and chemosensitizing effects of the Bcl-2 antisense oligodeoxynucleotide G3139 (oblimersen, Genasense) in NPC. For these studies, we used the C666-1 line, a stably infected NPC-derived line that co-expresses LMP-1 and Bcl-2. We have shown that G3139 treatment of C666-1 in vitro caused sequence dependent suppression of Bcl-2 protein, inhibition of cell growth and enhanced sensitivity to cisplatin (CDDP), as measured by increased antiproliferative and apoptotic effects. In vivo, G3139 treatment (25 mg/kg every 3 days x 5 doses) delayed engraftment and significantly inhibited growth of established C666-1 xenografts in SCID mice compared to control oligo-treated animals. However, G3139 alone did not prevent engraftment or cure established tumors in any animals. In contrast, G3139 treatment (25 mg/kg every 3 days x 5 starting on day 7) in combination with CDDP (8 mg/kg on day 14) completely abrogated tumor engraftment in 80% of animals compared to CDDP (0%) or CDDP + control oligo (0%). When treatment was delayed until tumor was established, G3139 in combination with CDDP significantly inhibited tumor growth compared to CDDP or CDDP + control oligo, and cured 69% animals with established tumors. No animals treated with G3139, CDDP or CDDP + control oligo were cured. Tumor burden and response to treatment correlated with EBV DNA load in serum, measured by real-time PCR. Western blots of tumor extracts obtained during oligo treatment showed that Bcl-2 levels were significantly decreased in G3139-treated animals. Our studies have demonstrated that the Bcl-2 antisense oligodeoxynucleotide, G3139, has proapoptotic effects in C666-1, and in combination with CDDP, is curative in C666-1 NPC xenograft tumors in vivo. The sequence-dependency of these effects is consistent with an antisense mechanism. These studies suggest that Bcl-2 may represent a biologically relevant target for the development of novel combinatorial therapies for NPC. PMID- 16477630 TI - Smoking is associated with a decrease of O6-alkylguanine-DNA alkyltransferase activity in bronchial epithelial cells. AB - O6-alkylguanine-DNA alkyltransferase (MGMT) represents the first line of defense against the toxic, mutagenic and carcinogenic effects of O6-alkylguanine adducts in DNA. These adducts mediate the biological activity from a series of alkylating agents, such as the tobacco-specific nitrosamines, believed to contribute to the carcinogenicity of tobacco smoke. There have been conflicting reports on the effects of smoking on MGMT activity in lung and other tissues. Here, we investigate MGMT activity in peripheral blood mononuclear cells (PBMC) and lung bronchial epithelial cells (BEC), extracted by lung brushings, from smokers and nonsmokers attending a bronchoscopy clinic. MGMT activity was significantly lower in BECs (geometric mean; 95% confidence interval 1.02; 0.86-1.20 fmol/microg DNA) than in PBMCs (7.86; 6.70-9.59 fmol/microg DNA; p < 0.001), suggesting that bronchial epithelia may be particularly sensitive to alkylation damage. More importantly our results indicate that activity in BECs is significantly decreased in samples from current smokers (0.71; 0.54-0.93 fmol/microg DNA) compared to nonsmokers (1.25; 1.03-1.51 fmol/microg DNA; p = 0.002). This could represent an important contribution to the carcinogenicity of tobacco smoke. PMID- 16477629 TI - Predicting 5-fluorouracil chemosensitivity of liver metastases from colorectal cancer using primary tumor specimens: three-gene expression model predicts clinical response. AB - We identified genes related to 5-fluorouracil (5-FU) sensitivity in colorectal cancer and utilized these genes for predicting the 5-FU sensitivity of liver metastases. Eighty-one candidate genes involved in 5-FU resistance in gastric and colon cancer cell lines were previously identified using a cDNA microarray. In this study, the mRNA expression levels of these 81 selected genes and the genes of 5-FU-related enzymes, including thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD) and orotate phosphoribosyltransferase (OPRT), were measured using real-time quantitative RT-PCR assays of surgically resected materials from primary colorectal tumors in 22 patients. Clinical responses were estimated by evaluating the effects of 5-FU-based hepatic artery injection (HAI) chemotherapy for synchronous liver metastases. Four genes (TNFRSF1B, SLC35F5, NAG-1 and OPRT) had significantly different expression profiles in 5-FU-nonresponding and responding tumors (p < 0.05). A "Response Index" system using three genes (TNFRSF1B, SLC35F5 and OPRT) was then developed using a discriminate analysis; the results were well correlated with the individual chemosensitivities. Among the 11 cases with positive scores in our response index, 9 achieved a reduction in their liver metastases after 5-FU-based chemotherapy, whereas only 1 of the 11 cases with negative scores responded well to chemotherapy. Our "Response Index" system, consisting of TNFRSF1B, SLC35F5 and OPRT, has great potential for predicting the efficacy of 5-FU-based chemotherapy against liver metastases from colorectal cancer. PMID- 16477631 TI - Familial risk of colon and rectal cancer in Iceland: evidence for different etiologic factors? AB - The aim of this study was to characterize the familial risk of colon and rectal cancer using 2 population-based registries in Iceland, the Icelandic Cancer Registry and a genealogy database. The standardized incidence ratio (SIR) was used to estimate the risk among relatives of colorectal cancer index cases diagnosed in Iceland over a 46-year period (1955-2000). The 2,770 colorectal cancer patients had 23,272 first-degree relatives. Among first-degree relatives, there was an increased risk of both colon (SIR 1.47, 95% confidence interval (CI) 1.34-1.62) and rectal cancer (SIR 1.24, 95% CI 1.04-1.47). An increased risk of colon cancer was observed among siblings of colon cancer patients (SIR 2.03, 95% CI 1.76-2.33), whereas no such increase was observed for parents or offspring. Furthermore, the risk of rectal cancer was only increased among brothers (SIR 2.46 95% CI 1.46-3.89) of rectal cancer patients and not among their sisters (SIR 1.0 95% CI 0.40-2.06). The added risk of colon cancer among first-degree relatives was independent of site of colon cancer in the proband. Our results confirm that family history of colorectal cancer is a risk factor for the disease. However, family history has a different association with colon cancer than with rectal cancer, suggesting that the 2 cancer types may have different etiologic factors. Our results have implications for colon and rectal cancer screening programs. PMID- 16477632 TI - Analysis of fumarate hydratase mutations in a population-based series of early onset uterine leiomyosarcoma patients. AB - Germline mutations in fumarate hydratase (FH) gene at 1q43 predispose to hereditary leiomyomatosis and renal cell cancer (HLRCC) syndrome. In HLRCC, the most common clinical features are leiomyomas of the skin and uterus, and in a subset of the families, renal cell cancer (RCC) and uterine leiomyosarcoma (ULMS) occur frequently at young age. This study was conducted to evaluate the possible contribution of FH mutations in a population-based series of early onset (< or = 45 years) ULMSs. Eighty-one cases were identified through the national cancer registry, and samples from 67 cases (83%) were available for FH mutation screening and analysis of allelic imbalance (AI) at the FH locus. Seventeen percent of tumors showed AI. In the mutation analysis, a novel missense mutation K424R was found. The mutation was also found from the patient's normal tissue. To study whether this variant has functional consequences, FH enzyme activity assay was performed in a cell model. The activity of the mutated protein was significantly reduced as compared to wild type (p = 0.009). This study shows that FH germline mutations can occur in seemingly nonsyndromic cases of ULMS (1/67, 1.5%). It appears that on the population level hereditary FH defects do play a role in pathogenesis of sporadic early onset ULMSs, albeit rarely. PMID- 16477633 TI - Rapid induction of apoptosis in B-cell lymphoma by functionally isolated human antibodies. AB - Novel panning and screening methodology was devised to isolate high affinity human recombinant scFv antibody fragments with functionally associated properties in B lymphoma cells. The approach was used to generate a panel of apoptosis inducing antibodies specific for antigens differentially expressed in B lymphoma vs. T leukaemia cells. The selections resulted in an antibody pool with near perfect selectivity (>99%) for the B lymphoma target cells. Randomly picked clones (72) revealed 7 unique antibody genotypes. Six of these rapidly induced apoptosis in target cells. Following the conversion to full IgGs, the antibodies were shown to be specific for HLA-DR/DP, the B-cell receptor mu chain and for CD54/ICAM-1. The latter receptor was not previously associated with apoptotic properties in B-cell lymphomas. Anti-ICAM-1 IgG induced apoptosis in a broad range of B lymphoma cell lines and were shown by immunohistochemistry to bind strongly to B lymphoma tissue obtained from 5 different B lymphoma patients. The recombinant IgG antibodies had affinities in the subnanomolar (0.3 nM) to nanomolar (3 nM) range. The described technology is generally applicable for the rapid isolation of high affinity human antibodies with specificity for differentially expressed cell surface receptors with intrinsic negative or positive signalling properties from naive phage libraries. PMID- 16477634 TI - Time trend analysis of the skin melanoma incidence of Finland from 1953 through 2003 including 16,414 cases. AB - Site-specific analyses of the skin melanoma incidence show marked differences between men and women by site and over time. The aim of our study was to analyze long-term population-based incidence time trends of skin melanoma in Finland over a period of more than 50 years, with special emphasis on sex- and subsite specific changes over time. We analyzed incidence data of the Finnish Cancer Registry from 1953 through 2003 including overall 16,414 cases. We calculated age standardized incidence rates per 100,000 person years using the European Standard Population. From 1953 through 2003, the incidence of skin melanoma increased from 1.5 to 12.8 per 100,000 among men and from 1.8 to 10.4 per 100,000 among women. Incidence rates showed a constant increase from 1953 through the mid of the 1980s. Thereafter, the rate of increase leveled off. The highest relative incidence increases occurred on the trunk among men and on the legs and hips among women. Within the skin area of the head, melanoma of the ear showed the highest relative increase among both men and women. Subsite-specific sex differences in the early registration period tended to become more pronounced in the most recent period. The highest body surface adjusted incidence rates occurred on the head. Only skin melanoma of the head showed an exponential age specific incidence pattern and the aetiology of these skin melanomas may differ from skin melanoma on other subsites. PMID- 16477635 TI - Treatment of squamous carcinoma in mice with a vaccine enriched for cells that induce immunity to squamous carcinoma--a new vaccination strategy. AB - We report a new vaccination strategy for squamous cell carcinoma (SCC). The vaccine was prepared by transfer of unfractionated DNA-fragments (25 kb) from squamous carcinoma cells (KLN205, DBA/2 origin (H-2(d))) into LM mouse fibroblasts (C3H/He origin; H-2(k)), a highly immunogenic cell line. To enhance their nonspecific immunogenic properties, the fibroblasts were modified before DNA transfer to secrete IL-2 and to express additional allogeneic MHC class I determinants. As the transferred DNA integrates into the genome of the recipient cells, and is replicated as the cells divide, sufficient DNA to prepare the vaccine could be obtained from as few as 10(7) squamous carcinoma cells (4 mm tumor). Since only a small proportion of the transfected cell-population was expected to have incorporated genes specifying antigens associated with the squamous carcinoma cells (TAA), we devised a novel approach to enrich the vaccine for cells that induce immunity to the SCC. Aliquots of the transfected population were divided into 10 small pools (initial inoculums = 1 x 10(3)). We reasoned that if the starting inoculums were sufficiently small, then the distribution of highly immunogenic and weakly immunogenic cells in each pool would not be the same. Cells from individual pools were allowed to increase in number. A portion of the expanded cell populations were maintained frozen/viable for later recovery. The remaining portions were used to immunize naive DBA/2 mice. Pools containing greater numbers of immunogenic cells were identified by 2 independent assays. Frozen aliquots of cells from the pool that stimulated immunity to the squamous carcinoma to the greatest extent were recovered and subdivided for additional rounds of immune selection. Enhanced immunity to squamous carcinoma mediated by CD8+ T cells was induced in tumor-bearing mice treated solely by immunization with the enriched cell-population. PMID- 16477636 TI - The prognostic impact of O6-Methylguanine-DNA Methyltransferase (MGMT) promotor hypermethylation in esophageal adenocarcinoma. AB - Promotor hypermethylation is a common event in human cancer. O6-Methylguanine-DNA Methyltransferase (MGMT) is a gene involved in DNA repair, which is methylated in a variety of cancer types. In colorectal cancer and lung cancer, hypermethylation of MGMT has been correlated with p53 mutation. In the present study, 132 samples of esophageal adenocarcinoma and 58 samples of normal esophageal tissue were investigated for MGMT hypermethylation status by methylation-specific real-time PCR and results were correlated to clinicopathological parameters, patient's survival, p53 mutation and expression of p53 protein and MGMT protein. In the carcinomas, hypermethylation of MGMT was found in 63.6% of cases and loss of MGMT protein expression in 48.5% of cases. Furthermore, MGMT hypermethylation was found in 5.7% of normal esophageal smooth muscle tissue, in 20.0% of esophageal squamous epithelium and in 61.5% of nonneoplastic Barrett's mucosa. In the carcinomas, hypermethylation of the MGMT gene was correlated with loss MGMT protein expression (p < 0.0001) and with high tumor differentiation (p = 0.0079). In contrast, no correlation between MGMT hypermethylation, Lauren's classification, WHO classification, tumor size, gender, age, pT category and pN category, and p53 status was found. Neither MGMT hypermethylation nor loss of MGMT protein expression was correlated with patient's survival. In conclusion, MGMT hypermethylation in esophageal adenocarcinoma is a frequent event that is associated with loss of MGMT protein expression but not with patient's outcome. PMID- 16477637 TI - Estrogen and progesterone receptor gene polymorphisms and sporadic breast cancer risk: a Spanish case-control study. AB - Estrogens, and to a lesser extent progesterones, influence the proliferation, differentiation and physiology of breast tissue as well as the development and progression of breast cancer. Genetic variants in the steroid hormone receptor genes ESR1 and PGR (belonging to the nuclear receptor superfamily) could therefore modify sporadic breast cancer susceptibility. Two studies have shown a protective effect associated with variants in ESR1 in 2 distinct populations. We studied 4 single nucleotide polymorphisms (SNPs) in ESR1 and 4 in PGR in 550 consecutive and unrelated sporadic Spanish breast cancer patients and 564 healthy Spanish controls. We observed a dominant protective effect for the S10S variant in ESR1, with an estimated odds ratio (OR) of 0.75 (95% CI = 0.58-0.97; p = 0.03) although functional studies did not show changes in the RNA stability. A small subset of individuals carried a haplotype combination that corroborates this protection. No other SNP considered in either gene was found to be associated with sporadic breast cancer. Our results obtained in a European population confirm the protective role of the S10S variant in ESR1, previously reported in an Asian and a European-American population. PMID- 16477638 TI - Diet, lifestyle and risk of K-ras mutation-positive and -negative colorectal adenomas. AB - K-ras mutation-positive (K-ras+) and -negative (K-ras-) colorectal adenomas may differ clinically and pathologically. As environmental compounds may cause mutations in the growth-related K-ras oncogene or affect clonal selection depending on mutational status, we evaluated whether the aetiology of K-ras+ and K-ras- adenomas differs. K-ras mutations in codons 12 and 13 were assessed in colorectal adenoma tissue (K-ras+: n = 81, K-ras-: n = 453). Dietary and lifestyle data were collected through questionnaires that were also administered to 709 polyp-free controls. Multiple logistic regression analyses showed that intake of vitamin B2 and monounsaturated fat were differently associated with risk of K-ras+ and K-ras- adenomas; vitamin B2 was inversely associated with K ras- (highest vs. lowest tertile: odds ratio (OR) = 0.70, 95% confidence interval (CI) = 0.50-0.97, p trend = 0.020), but not with K-ras+ adenomas, and a positive association with monounsaturated fat was confined to K-ras- adenomas (OR = 1.57, 95% CI = 1.06-2.34, p trend = 0.029). Besides, potential, not statistically significant, differences in risk arose because red meat was distinctly positively associated with K-ras+ adenomas (OR = 1.70, 95% CI = 0.94-3.09, p trend = 0.061); total dietary and polyunsaturated fat tended to be inversely associated with risk of K-ras+ but not of K-ras- adenomas; inverse associations with dairy products, calcium, protein and tea were confined to K-ras- adenomas, and smoking was more markedly positively associated with K-ras- adenomas. No differences in risk of K ras+ and K-ras- adenomas could be detected for other factors. In conclusion, dietary and lifestyle factors may influence risk of K-ras+ and K-ras- adenomas differently. However, epidemiological literature on diet, lifestyle and colorectal K-ras mutations is inconsistent. PMID- 16477639 TI - Biomarker analysis on breast ductal lavage cells in women with and without breast cancer. AB - Recent studies show that morphology based analysis of ductal lavage specimens failed to detect many cancers in women with breast cancer. Such an observation raises doubts about the potential role of ductal lavage in an individual's risk assessment and early detection of breast cancer. We hypothesize that biomarker based analysis using markers of malignancy field defects including DNA 5c exceeding rate (DNA 5cER) and G-actin might provide a more reliable test for breast cancer risk. The study was performed in 2 phases, the training and validation phase. For the training phase, 36 Chinese women were recruited (13 with breast cancer, 8 with intraductal papilloma and 15 with benign breast diseases). The validation phase included 10 women with cancer and 7 women without cancer. Ductal lavage samples were processed by the ThinPrep technique and evaluated by morphology followed by biomarker analysis using laser scan cytometry (LSC) for G-actin and DNA5cER. In the training phase, biomarker analysis was performed on the 67% (24 of 36) of samples that had over 100 epithelial cells. The sensitivity of DNA5cER was 90% with a specificity of 100%, and G-actin was 100% and 93%, respectively. By contrast, the sensitivity and specificity obtained by cytology alone were 67% and 93%, respectively. Similar results were obtained from the small validation study. Quantitative analysis of biomarkers for G-actin and DNA5cER is feasible and useful in distinguishing benign from malignant breast disease on archived ductal lavage slides. Further studies are warranted to determine the value of these biomarkers in prospective trials. PMID- 16477640 TI - Enhanced antitumor efficacy of telomerase-selective oncolytic adenoviral agent OBP-401 with docetaxel: preclinical evaluation of chemovirotherapy. AB - Oncolytic adenoviruses are being developed as novel anticancer therapeutics and currently undergoing clinical trials. We previously demonstrated that telomerase specific replication-competent adenovirus (Telomelysin: OBP-301), in which the human telomerase reverse transcriptase (hTERT) promoter regulates viral replication, efficiently killed human tumor cells. We further constructed OBP-401 (Telomelysin-GFP) that expresses the green fluorescent protein (GFP) reporter gene under the control of the cytomegalovirus promoter in the E3 region to monitor viral distribution. Here, we examined the feasibility of a single-agent therapy with OBP-401 as well as of combining OBP-401 with chemotherapeutic agents. Infection of OBP-401 alone or followed by the treatment of a chemotherapeutic drug, docetaxel (Taxotere), resulted in a profound in vitro cytotoxicity and GFP expression in various human cancer cell lines originating from different organs (lung, colon, esophagus, stomach, liver and prostate), although the magnitude of antitumor effect varied among the cell types. Other chemotherapeutic drugs such as vinorelbine (Navelbine) and SN38 (the potent active metabolite of irinotecan) combined with OBP-401 also inhibited the growth of human cancer cells. Quantitative real-time PCR analysis demonstrated that docetaxel did not affect viral replication. For in vivo evaluation, nu/nu mice xenografted with H1299 human lung tumor received intratumoral injection of OBP 401 and intraperitoneal administration of docetaxel. Analysis of growth of implanted tumors showed a significant, therapeutic synergism, although OBP-401 alone and docetaxel alone showed modest inhibition of tumor growth. Thus, OBP-401 in combination with docetaxel efficiently enhances the antitumor efficacy both in vitro and in vivo, and the outcome has important implications for tumor-specific oncolytic chemovirotherapies for human cancers. PMID- 16477641 TI - In Drosophila, don juan and don juan like encode proteins of the spermatid nucleus and the flagellum and both are regulated at the transcriptional level by the TAF II80 cannonball while translational repression is achieved by distinct elements. AB - The genes don juan (dj) and don juan like (djl) encode basic proteins expressed in the male germline. Both proteins show a similar expression pattern being localized in the sperm heads during chromatin condensation and along the flagella. Prematurely expressed Don Juan-eGFP and Myc-Don Juan Like localize to the cytoplasm of spermatocytes and in mitochondrial derivatives from the nebenkern stage onward suggesting that both proteins associate with the mitochondria along the flagella in elongated spermatids. Premature expression of Myc-Don Juan Like does not impair spermatogenesis where-as Don Juan-eGFP when prematurely expressed causes male sterility as spermatids fail to individualize. In spite of the sequence identity of 72% on the nucleotide level and 42% on the protein level, the presumptive promoter regions and the untranslated regions of the mRNA are diverged. Our in vivo analysis revealed that don juan and don juan like are transcriptionally and translationally controlled by distinct short cis regulatory regions. Transcription of don juan and don juan like depends on the male germ line specific TAF(II)80, Cannonball (Can). Translational repression elements for both mRNAs are localized in the 5' UTR and are capable to form distinct secondary structures in close proximity to the translational initiation codon. PMID- 16477642 TI - Development of a high-throughput membrane-array method for molecular diagnosis of circulating tumor cells in patients with gastric cancers. AB - Recently several noninvasive methods have been employed to detect circulating tumor cells (CTCs) in cancer patients. In this study, we have developed a highly sensitive, high-throughput colorimetric membrane-array method that was designed to detect a panel of mRNA markers including human telomerase reverse transcriptase (hTERT), cytrokeratin-19 (CK-19), carcinoembryonic antigen (CEA) and mucin 1 (MUC1) mRNA for the presence of CTCs in the peripheral blood of patients with gastric cancer (GC). Digoxigenin-labeled cDNA targets synthesized following total RNA isolation from peripheral blood samples of 64 GC patients and 80 healthy individuals were subjected to membrane-array hybridization. The results showed that membrane array could positively detect 5 cancer cells/ml of peripheral blood in GC cell-dilution experiments. The sensitivity, specificity and diagnostic accuracy for hTERT, CK-19, CEA and MUC1 mRNA ranged from 78.1% to 82.8%, 76.3% to 85% and 81.3% to 83.3%, respectively. Both CEA and MUC1 mRNA expression was correlated significantly with all malignant biological properties of GC, such as macroscopic type, depth of tumor invasion, lymph-node metastasis, TNM stage and coexisting distant metastasis (all p < 0.05). Using these 4 markers in combination, the sensitivity, specificity and diagnostic accuracy of membrane array were raised to 89.1%, 91.3% and 90.3%, respectively. The expression of all 4 mRNA markers was an independent predictor for postoperative recurrence/metastasis. GC patients with the expression of all the 4 mRNA markers showed a poorer survival rate than those without the expression of any 1 mRNA marker (p = 0.0223). These findings demonstrated that our membrane-array method could detect CTCs in the circulation of GC patients with considerably high sensitivity and specificity. The identification of CTCs in the peripheral blood may be useful in the auxiliary cancer diagnostics or postoperative surveillance of GC patients for recurrence/metastasis. PMID- 16477643 TI - Chemical cross-linking and mass spectrometry to map three-dimensional protein structures and protein-protein interactions. AB - Closely related to studying the function of a protein is the analysis of its three-dimensional structure and the identification of interaction sites with its binding partners. An alternative approach to the high-resolution methods for three-dimensional protein structure analysis, such as X-ray crystallography and NMR spectroscopy, consists of covalently connecting two functional groups of the protein(s) under investigation. The location of the created cross-links imposes a distance constraint on the location of the respective side chains and allows one to draw conclusions on the three-dimensional structure of the protein or a protein complex. Recently, chemical cross-linking of proteins has been combined with a mass spectrometric analysis of the created cross-linked products. This review article describes the most popular cross-linking reagents for protein structure analysis and gives an overview of the different available strategies that employ chemical cross-linking and different mass spectrometric techniques. The challenges for mass spectrometry caused by the enormous complexity of the cross-linking reaction mixtures are emphasized. The various approaches described in the literature to facilitate the mass spectrometric detection of cross-linked products as well as computer software for data analyses are reviewed. PMID- 16477644 TI - Inhibins are the major activin ligands expressed during early thymocyte development. AB - Activins are members of the transforming growth factor-beta (TGFbeta) superfamily, which regulate cell differentiation processes. Here we report the first quantitative analysis of the expression of Activin/Inhibin ligands, type I and II receptors, as well as Smad proteins in fetal (E14-E16) and adult thymic subpopulations. Our data showed that Alk4, ActRIIA, ActRIIB, and Smads 2, 3, and 4, are expressed in fetal thymus (E14 > E15 > E16) and in thymocytes from adult mice (mostly in the double negative [DN] subpopulation). Ligand expression analysis showed that betaA, betaB, and alpha subunits were mainly detected in thymic stromal cells. Interestingly, alpha subunits were expressed at much higher levels compared to betaA and betaB subunits, demonstrating for the first time the potential role of Inhibins as important mediators during early T cell development. Our data indicate that Activin/Inhibin signaling could regulate the process of thymus organogenesis and early thymocyte differentiation, as it has been demonstrated for other members of the TGF-beta superfamily. PMID- 16477645 TI - JAK/STAT signalling in Drosophila controls cell motility during germ cell migration. AB - The gonad is formed from two populations of cells originating at different locations: the primordial germ cells (PGCs), giving rise to either sperm or oocytes, and the somatic gonadal mesoderm precursors (SGPs), which support development of the gametes. Following the PGCs' migration during gastrulation, these two populations meet, forming the immature gonad. We present evidence that during embryonic development, the PGCs require the canonical JAK/STAT signalling cascade to migrate efficiently towards the SGPs. Loss of function for any element of the JAK/STAT pathway causes frequent germ cell mislocalisation. We have found that wild-type germ cells produce filopodia while they migrate through the mesoderm towards the gonad. Our observations suggest that PGCs use filopodia to migrate and to keep contact with each other. Interestingly, activation of the JAK/STAT pathway is required for these filopodia to form, and ectopic JAK/STAT activation enhances their formation. PMID- 16477646 TI - Immunohistochemical study of the distribution of sodium-dependent vitamin C transporters in adult rat brain. AB - Sodium-dependent vitamin C transporters (SVCTs) is known to transport the reduced form of ascorbic acid into the cell, whereas the oxidized form of vitamin C (VC) is moved through a facilitative sugar transporter, such as glucose transporter (GLUT). With regard to the distribution of SVCT1 and -2 within the various organs, they were reported to be expressed in different types of cells. Especially in the central nervous system, only SVCT2 mRNA was expressed mainly in neurons and some types of neuroglial cells. However, data on the expression of SVCT proteins in the brain are scant. Therefore, we tried to develop comprehensive data on the distribution of SVCT proteins in adult rat brain by using immunohistochemical techniques for the first time. In our study, SVCT2 immunoreactivities (IRs) were intensely localized in the neurons of cerebral cortex, hippocampus, and Purkinje cells of cerebellum, and much weaker SVCT2 IRs were found in the other brain regions. Judging from double-immunohistochemical data, most of the cells expressing SVCT2 IRs were likely to be neurons or microglia, even though the cells in choroids plexus or ependymal cells around the ventricles also exhibited SVCT2 IRs. Complete mapping of the distribution of SVCT2 IRs was available by using a semiquantitative method. The subcellular localization of SVCT proteins is necessary for understanding the exact role of the protein, so the current overall mapping of SVCT IRs in the rat brain could be the basis for further studies on related subjects. PMID- 16477647 TI - Cloning and regulation of the vertebrate homologue of lin-41 that functions as a heterochronic gene in Caenorhabditis elegans. AB - The Caenorhabditis elegans gene lin-41 is the one of the heterochronic genes that regulate the timing of many developmental events. MicroRNA let-7 negatively regulates the expression of lin-41 through RNA-RNA interaction on its 3' untranslated region (UTR). Here, we report the isolation of chick and mouse homologues of lin-41 that encode the RBCC-NHL family protein and their expression patterns. C. elegans lin-41 is one of the RBCC-NHL families and the predicted amino acid sequences of isolated two genes encode the same family proteins. Chick and mouse lin-41 expression was also observed in developing limb buds, branchial arches, and tail buds. The 3'UTRs of the mouse and chick lin-41 genes contain multiple let-7 complementary sites. Using luciferase assay, we showed that lin-41 expression can be regulated through let-7 complementary sites. PMID- 16477649 TI - Evolutionary coupling of structural and functional sequence information in the intracellular lipid-binding protein family. AB - We have mined the evolutionary record for the large family of intracellular lipid binding proteins (iLBPs) by calculating the statistical coupling of residue variations in a multiple sequence alignment using methods developed by Ranganathan and coworkers (Lockless and Ranganathan, Science 1999:286;295-299). The 213 sequences analyzed have a wide range of ligand-binding functions as well as highly divergent phylogenetic origins, assuring broad sampling of sequence space. Emerging from this analysis were two major clusters of coupled residues, which when mapped onto the structure of a representative iLBP under study in our laboratory, cellular retinoic-acid binding protein I, are largely contiguous and provide useful points of comparison to available data for the folding of this protein. One cluster comprises a predominantly hydrophobic core away from the ligand-binding site and likely represents key structural information for the iLBP fold. The other cluster includes the portal region where ligand enters its binding site, regions of the ligand-binding cavity, and the region where the 10 stranded beta-barrel characteristic of this family closes (between strands 1' and 10). Linkages between these two clusters suggest that evolutionary pressures on this family constrain structural and functional sequence information in an interdependent fashion. The necessity of the structure to wrap around a hydrophobic ligand confounds the typical sequestration of hydrophobic side chains. Additionally, ligand entry and exit require these structures to have a capacity for specific conformational change during binding and release. We conclude that an essential and structurally apparent separation of local and global sequence information is conserved throughout the iLBP family. PMID- 16477648 TI - Developmental expression patterns of Tbx1, Tbx2, Tbx5, and Tbx20 in Xenopus tropicalis. AB - T-box genes have diverse functions during embryogenesis and are implicated in several human congenital disorders. Here, we report the identification, sequence analysis, and developmental expression patterns of four members of the T-box gene family in the diploid frog Xenopus tropicalis. These four genes-Tbx1, Tbx2, Tbx5, and Tbx20-have been shown to influence cardiac development in a variety of organisms, in addition to their individual roles in regulating other aspects of embryonic development. Our results highlight the high degree of evolutionary conservation between orthologs of these genes in X. tropicalis and other vertebrates, both at the molecular level and in their developmental expression patterns, and also identify novel features of their expression. Thus, X. tropicalis represents a potentially valuable vertebrate model in which to further investigate the functions of these genes through genetic approaches. PMID- 16477650 TI - Regulation by GD3 of the proinflammatory response of microglia mediated by interleukin-15. AB - The interleukin (IL)-15-dependent immune responses of murine microglia were strongly affected by low concentrations of the ganglioside GD3. The ganglioside binding to IL-15 inhibited the proinflammatory effects of the cytokine, reducing IL-15-dependent T-cell proliferation as well as mRNA expression for IL-15Ralpha, p65, and NFATc2 in the N13 murine microglial cell line. Treatment of primary murine microglial cultures with GD3 abolished IL-15 production, without affecting cellular viability, but decreased the production of nitric oxide, a direct sensor of inflammation and nuclear factor-kappaB activity. We conclude that low doses of GD3 could inhibit specific proinflammatory mechanisms and modulate the inflammatory environment, leading to a less reactive scene. Microglial cells are one of the main actors in the inflammatory events that follow CNS trauma or an autoimmune disease episode, modulating the internal production of cytokines, growth factors, and other homeostatic molecules that may determine the evolution and outcome of tissue damage. Proinflammatory cytokines have a relevant role in the initial events, and modulation of their activity by gangliosides could cut down their harmful effects and interfere with invasion of the CNS by peripheral immune cells. The antiinflammatory properties of GD3 could be significant in the treatment of pain subsequent to CNS damage. PMID- 16477651 TI - Identification and expression profiling of 10 novel spermatid expressed CYPT genes. AB - To identify candidate genes for poor sperm morphology, we have screened for genes expressed during spermiogenesis. We identified 10 new members of the cysteine rich perinuclear theca (CYPT) family showing that this family contains at least 15 members, which also includes the casein kinase II target genes. Based on similarity the CYPT sequences could be divided into two groups, Cypt1-10 and the novel members Cypt12-15. The 5'-end of the CYPT family is highly similar to exon1A and part of the first intron of Zfy2. Seven CYPT genes mapped to the X chromosome; six contained an intron and one was intron-less. One CYPT gene mapped to chromosome 3 and one mapped to chromosome 9 which were both intron-less. The upstream region of the CYPT family and Zfy2 genes is conserved. For some the conservation extended over a large region, however, only about 150 nucleotides is conserved among all CYPT members and Zfy2. Nevertheless, the short conserved promoter leads to essentially identical expression profiles for the CYPT family members and Zfy2, which was clearly different from the profile of Zfy1. Expression of the CYPT family and Zfy2 preceded the expression of other spermatid specific genes such as the transition proteins and the protamines. In situ hybridization revealed a low expression in pachytene spermatocytes from stages IX X followed by a strong upregulation in spermatids from stage VI with maximum expression in spermatids in stages VII-VIII. The CYPT family may function in the remodeling of the spermatid nucleus before condensation of the DNA. PMID- 16477653 TI - Lung inflammation in preterm infants with respiratory distress syndrome: effects of ventilation with different tidal volumes. AB - Ventilation with an inappropriate tidal volume (Vt) triggers lung inflammation, an important predisposing factor of bronchopulmonary dysplasia. It still remains uncertain what the appropriate starting target Vt should be during the acute phase of respiratory distress syndrome (RDS). Our aim was to evaluate lung inflammation in preterm infants undergoing synchronized intermittent positive pressure ventilation (SIPPV) with two different tidal volumes Vt during the acute phase of RDS. Thirty preterm infants (gestational age, 25-32 weeks) with acute RDS were randomly assigned to be ventilated with Vt = 5 ml/kg (n = 15) or Vt = 3 ml/kg (n = 15). Proinflammatory cytokines (interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor (TNF)-alpha) were determined in the tracheal aspirate on days 1, 3, and 7 of life. IL-8 and TNF-alpha levels collected on day 7 were significantly higher (P < 0.05), and mechanical ventilation lasted longer in the group with Vt = 3 ml/kg (16.8 +/- 4 vs. 9.2 +/- 4 days; P = 0.05). In conclusion, our data show significantly higher lung inflammation in preterm infants ventilated with Vt = 3 ml/kg, suggesting a role for Vt = 5 ml/kg in reducing both inflammatory response during the acute phase of RDS and the length of ventilation. Whether the use of this starting Vt prevents bronchopulmonary dysplasia requires further study. PMID- 16477652 TI - Regulation of human neural precursor cells by laminin and integrins. AB - Deciphering the factors that regulate human neural stem cells will greatly aid in their use as models of development and as therapeutic agents. The extracellular matrix (ECM) is a component of stem cell niches in vivo and regulates multiple functions in diverse cell types, yet little is known about its effects on human neural stem/precursor cells (NSPCs). We therefore plated human NSPCs on four different substrates (poly-L-ornithine, fibronectin, laminin, and matrigel) and compared their responses with those of mouse NSPCs. Compared with the other substrates, laminin matrices enhanced NSPC migration, expansion, differentiation into neurons and astrocytes, and elongation of neurites from NSPC-derived neurons. Laminin had a similar spectrum of effects on both human and mouse cells, highlighting the evolutionary conservation of NSPC regulation by this component of the ECM. Flow cytometry revealed that human NSPCs express on their cell surfaces the laminin-binding integrins alpha3, alpha6, alpha7, beta1, and beta4, and function-blocking antibodies to the alpha6 subunit confirmed a role for integrins in laminin-dependent migration of human NSPCs. These results define laminin and its integrin receptors as key regulators of human NSPCs. PMID- 16477654 TI - Mitochondrial dysfunction in Brooks-Wisniewski-Brown syndrome. AB - Brooks, Wisniewski, and Brown described a familial presentation of severe developmental retardation, speech delay, static encephalopathy with atrophic hydrocephalus, microcephaly, progressive spastic diplegia, a characteristic facial appearance, optic atrophy, and growth retardation associated with hypoplastic corpus callosum in one of the patients. The authors postulated a distinct X-linked mental retardation syndrome. Later on a similar phenotype was observed in three male siblings with an early lethal outcome. Here we describe three patients with several overlapping features and a progressive neurological picture presenting with a significantly compromised mitochondrial oxidative phosphorylation measured in a fresh muscle biopsy. Neurological deterioration is a commonly observed feature in mitochondrial disorders. Based on the unique combination of the clinical symptoms, we suggest that our patients have the Brooks-Wisniewski-Brown syndrome. PMID- 16477655 TI - Pediatric Churg-Strauss syndrome in Mexico. AB - Churg-Strauss syndrome (CSS) is one of the rarest forms of vasculitis, and very rarely presents in the pediatric population. We present two cases of childhood CSS, both with hepatic and cardiac involvement. To our knowledge, these are the first two cases of CSS in the pediatric population described in Mexico. PMID- 16477656 TI - Effect of body mass index on response to methacholine bronchial provocation in healthy and asthmatic adolescents. AB - A linkage between airway hyperreactivity and obesity could partly explain the prevalence of obesity in asthmatics. To test for such a linkage, we analyzed body mass index (BMI), pulmonary function, methacholine bronchial provocation, and asthma severity scores in 216 adolescents (aged 12-18 years), of whom 82 were healthy and 134 were asthmatic. Methacholine provocations in a subgroup of 36 subjects (healthy and asthmatic) enabled us to examine the effects of BMI on dynamic hyperinflation and ventilatory indices during induced bronchospasm. Age- and gender-specific BMI was higher in asthmatics (74 +/- 24%) compared to healthy subjects (61 +/- 28%, P < 0.002). General linear model analysis, in which baseline spirometric results were adjusted for gender, age, race, and height, showed opposing effects of BMI on expiratory flow in controls and asthmatics (P < 0.05), i.e., forced expired volume in 1 sec increased with BMI in controls (P < 0.02), but forced expiratory flow (FEF)(25-75%) decreased with BMI in asthmatics (P < 0.05). However, linear regression analysis showed no effect of BMI on the provocation dose for methacholine (PD(20)) in either controls or asthmatics, and there was no effect of BMI on asthma severity scores. Overweight (BMI >85th percentile) and nonoverweight subjects had similar degrees of dynamic hyperinflation during positive provocations, but overweight subjects had greater decreases in mean inspiratory flow (mean, 28% vs. 9%, P < 0.05). We conclude that our measurements support a relationship between overweight and baseline flow limitation, rather than a relationship between overweight and airways hyperreactivity, in the linkage between overweight and asthma during adolescence. PMID- 16477657 TI - Is significant cystic fibrosis-related liver disease a risk factor in the development of bone mineralization abnormalities? AB - In order to assess the effects of significant cystic fibrosis-related liver disease (CFLD) on bone health, we compared the bone mineral status of older children and adolescents with CFLD to those with cystic fibrosis (CF) alone. Thirteen children (age range, 10-19 years) from our clinical CF services were identified with significant CFLD (9 of these 13 patients had clinical and radiological evidence of portal hypertension). This cohort was then matched by age, gender, and anthropometric measurements with equal numbers of patients with CF alone. All patients had a dual-energy X-ray absorptiometry (DEXA) scan to determine bone mineral content (BMC), bone area (BA), bone mineral density (BMD), and bone mineral apparent density (BMAD) in the region of the lumbar spine. Blood was drawn to determine serum vitamin A, D, E, and K status and liver function tests. The best forced expired volume in 1 sec (FEV1) for each patient in the 12 months around the time of the scan was also documented. Patients with CFLD had slightly worse FEV1 (82 +/- 20% vs. 91 +/- 16%, P = 0.05) and significantly higher alanine aminotransferase (65.5 +/- 35 IU/l vs. 30 +/- 20 IU/l, P = 0.01) than those with CF alone. The mean lumbar spine BA, BMC, BMD, and BMAD were not different between children with CFLD and CF. In conclusion, the presence of significant liver disease in children with CF does not appear to be an additional risk factor for the development of abnormal bone mineralization. PMID- 16477658 TI - Factors influencing gender differences in the diagnosis and treatment of asthma in childhood: the Tucson Children's Respiratory Study. AB - Studies identified gender differences in diagnosed asthma, but the extent to which they can be attributed to differences in symptom experience and frequency rather than factors influencing diagnosis has not been established. We investigated prevalence of, and consultation for, asthma symptoms, as well as diagnosis and treatment in 533 boys and 556 girls enrolled in the Tucson Children's Respiratory Study, a population-based birth-cohort study. Questionnaires regarding respiratory symptoms and diagnoses were obtained at ages 2, 3, 6, 8, 11, 13, 16, and 18 years. Boys were significantly more likely than girls to experience both wheeze and frequent wheeze most years in the first decade of life. However, girls with symptoms were less likely than boys to see a physician (74.1% vs. 83.4%, P < 0.001) and to be labeled as having asthma (43.3% vs. 53.8%, P < 0.009), even after adjusting for symptom frequency. A difference in symptom presentation also appeared to influence diagnosis: nocturnal cough without frequent wheeze was more prevalent among girls, and was associated with reduced diagnosis of asthma. Among subjects who consulted a physician for wheeze, boys were significantly more likely than girls to have taken medication (81.5% vs. 73.5%, P < 0.01). The lag time between age at first wheeze and first use of medication among those consulting a physician for wheeze or asthma was greater for girls, especially among subjects with frequent wheeze (2.8 vs. 1.6 years, P < 0.005). These findings indicate that gender differences in the diagnosis and treatment of asthma cannot be explained completely by differences in symptom prevalence and frequency. PMID- 16477659 TI - Comparative study of biological nutrient removal (BNR) processes with sedimentation and membrane-based separation. AB - A membrane-enhanced biological phosphorus removal (MEBPR) process was operated in parallel with a conventional EBPR (CEBPR) process under challenging operating conditions to uncover fundamental differences in their ability to remove chemical oxygen demand (COD), nitrogen (N), and phosphorus (P) from municipal wastewater. Both systems exhibited the same potential to achieve excellent soluble-P removal when a favorable COD to P ratio was maintained in the influent. The MEBPR train generated a superior effluent quality when measured as total P. The CEBPR effluent contained significantly lower levels of nitrates due to the extra denitrification occurring in the sludge blanket of the secondary clarifier. The observed sludge yield in the MEBPR system was estimated to be between 0.23 and 0.28 g VSS/g COD, and this was 15% lower than the CEBPR sludge yield. When the influent volatile fatty acids (VFAs) became limiting, the CEBPR train exhibited better performance in the removal of soluble-P, due to the higher observed sludge yield and an overall greater denitrification activity that led to a more efficient use of VFAs in the anaerobic zone. After experiencing a severe deterioration of the biological P activity in both processes, the MEBPR train exhibited faster recovery than the CEBPR side. In this experimental work, it was demonstrated that an MEBPR process can sustain long-term satisfactory bio-P performance at HRTs as low as 7 h. However, the lower sludge yield and the reduced denitrification capacity are two important factors that impact the design of high rate membrane-assisted biological nutrient removal (BNR) processes. PMID- 16477660 TI - Cyclodextrins micrometric powders obtained by supercritical fluid processing. AB - Supercritical fluid technology offers the possibility to produce dry powder formulations of biocompatible materials, overcoming the drawbacks of classical micronization processes. In this work, Supercritical Assisted Atomization (SAA) has been used to micronize alpha-cyclodextrin (alpha-CD) and hydroxypropyl-beta cyclodextrin (HP-beta-CD). Some process parameters, such as precipitation temperature and solute concentration in the liquid solution, have been studied to evaluate their influence on morphology and size of precipitated particles. Cyclodextrins (CDs) micronization has been successful: well-defined spherical microparticles of alpha-CD and HP-beta-CD have been produced. Particle size analysis revealed that sharp distributions have been obtained: 95% of particles have diameters ranging between 0.1 and 5 microm for both CDs. X-ray and DSC analyses have been also performed to investigate CDs modifications induced by SAA processing: amorphous particles have been obtained in both cases, whereas raw alpha-CD was crystalline and raw HP-beta-CD was amorphous. PMID- 16477661 TI - Community structures and activities of nitrifying and denitrifying bacteria in industrial wastewater-treating biofilms. AB - The bacterial community structure, in situ spatial distributions and activities of nitrifying and denitrifying bacteria in biofilms treating industrial wastewater were investigated by combination of the 16S rRNA gene clone analysis, fluorescence in situ hybridization (FISH) and microelectrodes. These results were compared with the nitrogen removal capacity of the industrial wastewater treatment plant (IWTP). Both nitrification and denitrification occurred in the primary denitrification (PD) tank and denitrification occurred in the secondary denitrification (SD) tank. In contrast, nitrification and denitrification rates were very low in the nitrification (N) tank. 16S rRNA gene clone sequence analysis revealed that the bacteria affiliated with Alphaproteobacteria, followed by Betaproteobacteria, were numerically important microbial groups in three tanks. The many clones affiliated with Alphaproteobacteria were closely related to the denitrifying bacteria (e.g., Hyphomicrobium spp., Rhodopseudomonas palustris, and Rhodobacter spp.). In addition, Methylophilus leisingeri affiliated with Betaproteobacteria, which favorably utilized methanol, was detected only in the SD-tank to which methanol was added. Nitrosomonas europaea and Nitrosomonas marina were detected as the ammonia-oxidizing bacteria affiliated with Betaproteobacteria throughout this plant, although the dominant species of them was different among three tanks. Nitrifying bacteria were mainly detected in the upper parts of the PD-biofilm whereas their populations were low in the upper parts of the N-biofilm. The presence of denitrifying bacteria affiliated with Hyphomicrobium spp. in SD- and N-biofilms was verified by FISH analysis. Microelectrode measurements showed that the nitrifying bacteria present in the N- and PD-biofilms were active and the bacteria present in the SD-biofilm could denitrify. PMID- 16477662 TI - Optical method for long-term and large-scale monitoring of spatial biofilm development. AB - A method was developed that allows biofilm monitoring on the square centimeter scale over extended periods of time. The method is based on image acquisition using a desktop scanner and subsequent image analysis. It was shown that results from grey level analysis are highly correlated with physical properties of the biofilm like average biomass and biofilm thickness. The scanner method was applied to monitor overall biofilm growth, detachment, and surface roughness during two 3 and 4 week long experiments. Two significantly different growth dynamics during the biofilm development could be identified, depending on the biofilm history. Surface roughness on transects in flow direction was always higher than on transects perpendicular to the flow, reflecting the anisotropic characteristics of biofilms growing in a flow field. PMID- 16477663 TI - Towards the determination of the absolute configuration of complex molecular systems: matrix isolation vibrational circular dichroism study of (R)-2-amino-1 propanol. PMID- 16477665 TI - Understanding the failure of direct C-C coupling in the zeolite-catalyzed methanol-to-olefin process. PMID- 16477664 TI - Large-scale synthesis of H-antigen oligosaccharides by expressing Helicobacter pylori alpha1,2-fucosyltransferase in metabolically engineered Escherichia coli cells. PMID- 16477666 TI - Gold(I)-catalyzed intramolecular hydroamination of alkenyl carbamates. PMID- 16477667 TI - Atomic-force-microscopy imaging and molecular-recognition-force microscopy of recrystallized heterotetramers comprising an S-layer-streptavidin fusion protein. PMID- 16477668 TI - The properties of weak and strong dihydrogen-bonded D-H...H-A complexes. AB - The properties of six dihydrogen-bonded (DHB) dimers with the BeH2 molecule as a proton acceptor were calculated by MP2, CCSD(T) and B3LYP methods. The structural, energetic and spectroscopic parameters are presented and analyzed in terms of their possible correlation with the interaction energy and the intermolecular H...H separation. The symmetry-adapted perturbation theory (SAPT) calculations were performed to gain more insight into the nature of the H...H interactions. The studied complexes are divided into three groups based on the calculated intermolecular distances and the interaction energies which range from approximately -1 to -42 kJ mol(-1). The analysis of the interaction energy components indicates that, in contrast to conventional hydrogen bonds, the induction energy is the most important term in the BeH2NH4+ complex. On the other hand, there is no sharp boundary between the DHB complexes classified as hydrogen bonded and van der Waals systems. The complexation-induced changes in vibrational frequencies and in proton shielding constants show a relationship with the interaction energy. The values of the 2hJXH and 3hJBeX coupling constants correlate well with the interaction energy and with the intermolecular distance. PMID- 16477669 TI - Electron traps on oxide surfaces: (H+)(e-) pairs stabilized on the surface of 17O enriched CaO. AB - (H+)(e-) pairs generated at the surface of polycrystalline CaO are analyzed for the first time in terms of the interaction of the unpaired electron spin with the nuclear spin of the 17O anions of the surface. CaO crystals enriched in the 17O isotope are prepared and the corresponding hyperfine coupling constants are measured in electron paramagentic resonance (EPR) spectra. The results are analyzed on the basis of cluster model density functional theory calculations. The computed hyperfine coupling constants for (H+)(e-) pairs formed on the edge, corner, and reverse corner sites of the CaO surface allow a tentative assignment of two observed spectral features to specific morphological surface sites. PMID- 16477670 TI - Comparison of promoter region constructs for in vivo intramuscular expression. AB - BACKGROUND: High transgene expression is generally expected after gene transfer. However, different level, kinetics and localization of expression might be needed for relevant therapeutic applications. Former studies have compared various promoter regions driving gene expression leading to conflicting results. In the present work, two promoter families have been compared using the efficient in vivo intramuscular electrotransfer technique. METHODS: Three promoter regions were constructed by associating the strong ubiquitous cytomegalovirus (CMV) enhancer-promoter to its homologous intron A or to a heterologous intron, or to a hybrid intron. Promoter regions derived from the muscle creatine kinase (MCK) promoter were also studied. The expression of the same transgene (SeAP or neurotrophin-3) under control of these different promoters was compared after plasmid electrotransfer in mouse tibialis-cranialis skeletal muscle. RESULTS: Heterologous intron association to the CMV promoter did not modify gene expression kinetics nor increase gene expression level. Usefulness of intron A or hybrid intron association to the CMV promoter depended on the gene. The various MCK promoters drove efficient gene expression but lower than that obtained with the CMV promoter. Furthermore, peak value was reached earlier with MCK promoter regions (14 days). CONCLUSION: For applications of gene transfer restricted to skeletal muscle, the MCK promoter or a MCK promoter variant would be a promising alternative to the CMV promoter. Indeed, it has been demonstrated that the use of MCK promoter limits humoral and cell-mediated immune responses. Furthermore, the MCK promoter decreases the initial expression peak that may be detrimental, drives a sustained gene expression, and improves gene transfer safety. PMID- 16477671 TI - Revisiting the calculation of (13)C chemical shift tensors in cadmium acetate dihydrate with EIM and EIM/cluster methods. AB - The chemical shift tensors of the acetate anions in cadmium acetate dihydrate are calculated using a cluster approach, the embedded ion method (EIM), and a combination of the two in the EIM/cluster method. The results of these calculations are compared with those completed on the isolated acetate anion and show the need for the inclusion of intermolecular interactions. The RMS difference between experiment and theory improves from over 60 ppm when the calculation is completed on an isolated anion, to below 10 ppm when interactions to nearby atoms are included. The best cluster model includes three cadmium acetate dihydrate and gives an RMS result of 4.4 ppm. The EIM method, which uses point charges to account for the intermolecular effects, achieves an RMS of 7.7 ppm on individual anions alone. A combination of the two, the EIM/cluster method, shows that the only necessary atom to explicitly add is the nearest cadmium; this addition results in an RMS of 4.1 ppm. These results are also discussed in terms of the computational cost of the different calculations. PMID- 16477672 TI - Modeling the (13)C chemical-shift tensor in organic single crystals by quantum mechanical methods: finite basis set effects. AB - The influence of using finite basis sets to calculate (13)C magnetic shieldings were explored using the Hartree-Fock and the B3LYP hybrid density functional methods. The shielding values were compared in a linear least-squares fashion for a test group of 102 (13)C complete chemical-shift tensors determined from 14 organic single crystals. Pople's basis sets allow for the addition of polarization and diffuse functions in a straightforward way, allowing the examination of 81 combinations at the double and triple zeta level. Dunning's correlation-consistent basis sets were explored as well. The errors associated with predicting the shielding values were found to be largely systematic as revealed by the analysis of the determined regression parameters between calculated chemical shieldings and experimental chemical shifts. Expansion of the basis set leads to a convergence of these regression parameters to their ideal values. The random errors, however, do not decrease by employing larger basis sets; therefore, given the appropriate regression parameters, a small basis description such as 3-21G can be adequate in predicting the relative magnetic shielding values, i.e. the chemical shifts. Furthermore, in certain cases the inclusion of unbalanced diffuse and polarization functions can significantly degrade the predicted shielding rmsd. Unless employed carefully, these functions do not justify their computational expense. The chemical-shift distance is used to evaluate shielding predictions in individual tensor components. The analysis of the chemical-shift's distance between calculated and experimental data indicates an orientational dependence on the magnitude of errors and suggests the use of the shift anisotropy as a useful fiduciary mark to optimize model chemistries for magnetic-shielding calculations. PMID- 16477673 TI - Intermolecular shielding from molecular magnetic susceptibility. A new view of intermolecular ring current effects. AB - This paper presents calculations of the NICS (nuclear independent chemical shieldings) in a rectangular grid surrounding the molecules of benzene, naphthalene and coronene. Using the relationship between calculated NICS and the induced magnetic field, the calculated NICS are used to predict intermolecular effects due to molecular magnetic susceptibility or ring current effects. As expected from approximate ring current models, these intermolecular shielding effects are concentrated along the direction perpendicular to the molecular plane and they approach asymptotically to a dipolar functional dependence, i.e. (1-3 cos(2)theta)/r(3)). The deviations from the dipolar functional form require that the calculations of these intermolecular effects be done using a suitable interpolation scheme of the NICS calculated on the grid. The analysis of the NICS tensor components shows that these intermolecular shielding effects should be primarily expected on shielding components of the neighboring molecules nuclei, which are perpendicular to the molecular plane of the aromatic compound generating the induced field. The analysis of the calculated NICS along the series benzene, naphthalene and coronene shows that these intermolecular effects increase monotonically with the number of aromatic rings. PMID- 16477675 TI - Calculating protein structures directly from anisotropic spin interaction constraints. AB - Protein structure determination by solid-state NMR of aligned samples relies on the fundamental characteristics of the anisotropic nuclear spin interactions present in isotopically labeled proteins. Progress in the implementation of algorithms that calculate protein structures from the orientational constraints in the chemical shift and heteronuclear dipolar coupling interactions is described using both simulated and experimental data. PMID- 16477674 TI - Cognitive function, fatigue, and menopausal symptoms in breast cancer patients receiving adjuvant chemotherapy: evaluation with patient interview after formal assessment. AB - BACKGROUND: Women who receive adjuvant chemotherapy for breast cancer report fatigue, menopausal symptoms and cognitive problems. Here we compare assessment of these symptoms using self-report questionnaires and a researcher-administered screen of cognitive function with the experience of women as revealed in a semi structured interview. METHODS: Twenty-one women who were receiving adjuvant chemotherapy completed the Functional Assessment of Cancer Treatment-General (FACT-G) self-report questionnaire, and sub-scales for fatigue (FACT-F) and endocrine symptoms (FACT-ES). They were evaluated for cognitive dysfunction using the High Sensitivity Cognitive Screen (HSCS). They then completed a semi structured interview, which explored the nature and severity of these symptoms and their impact on daily function. RESULTS: All patients experienced fatigue and most had menopausal symptoms. There was reasonable correlation of findings in the interview with FACT-F and FACT-ES scores. The HSCS revealed fewer problems than were reported by patients, and correlated with patient experience only for the domain of memory. Most patients noted adverse changes in other cognitive domains, especially concentration, with substantial effects on every-day function. CONCLUSIONS: Women receiving adjuvant chemotherapy for breast cancer have substantial problems with fatigue, menopausal symptoms and cognitive changes. Formal tests such as the HSCS may fail to adequately capture the perceived impact of symptoms. PMID- 16477676 TI - Carbon-13 chemical shift anisotropy in DNA bases from field dependence of solution NMR relaxation rates. AB - Knowledge of (13)C chemical shift anisotropy (CSA) in nucleotide bases is important for the interpretation of solution-state NMR relaxation data in terms of local dynamic properties of DNA and RNA. Accurate knowledge of the CSA becomes particularly important at high magnetic fields, prerequisite for adequate spectral resolution in larger oligonucleotides. Measurement of (13)C relaxation rates of protonated carbons in the bases of the so-called Dickerson dodecamer, d(CGCGAATTCGCG)(2), at 500 and 800 MHz (1)H frequency, together with the previously characterized structure and diffusion tensor yields CSA values for C5 in C, C6 in C and T, C8 in A and G, and C2 in A that are closest to values previously reported on the basis of solid-state FIREMAT NMR measurements, and mostly larger than values obtained by in vacuo DFT calculations. Owing to the noncollinearity of dipolar and CSA interactions, interpretation of the NMR relaxation rates is particularly sensitive to anisotropy of rotational diffusion, and use of isotropic diffusion models can result in considerable errors. PMID- 16477678 TI - (1)H/(29)Si cross-polarization NMR experiments of silica-reinforced polydimethylsiloxane elastomers: probing the polymer-filler interface. AB - Polydimethylsiloxane (PDMS) elastomers reinforced with fumed silica exhibit unusual strength characteristics that are necessary for their designed applications. The microscopic details of the surface interaction between the polymer and silica are not well characterized. (1)H/(29)Si cross-polarization (cp) experiments are used to characterize cured and uncured samples of Dow Corning silastic 745. Changes to the cp dynamics upon curing are evident by the variation in peak intensities in the variable contact-time spectra of the two samples. Estimates of the cp relaxation parameters are reported for the cured sample. Additional information can be obtained by expanding the (1)H/(29)Si cp to a two-dimensional heteronuclear correlation experiment. Dramatic differences between the cured and uncured (1)H/(29)Si HetCor spectra are observed that are not visible in the 1D spectra. These changes can be rationalized as a dehydration of the silica surface and an increased hardening of the polymer after the curing process. Furthermore, isolation of the NMR signal corresponding to nuclei at or near the polymer-filler interface may be achieved in the 2D (1)H/(29)Si HetCor spectrum. PMID- 16477677 TI - Demographic and clinical predictors of preparatory grief in a sample of advanced cancer patients. AB - BACKGROUND: Preparatory grief encompasses grief for losses that have already occurred, are currently being experienced, and losses that will or might ensue in the future after the death, as a consequence of it. AIM: To examine the relative contribution of demographic and clinical variables in predicting cancer patients' preparatory grief as recorded from the Preparatory Grief in Advanced Cancer Patients (PGAC) scale. Moreover, researchers were interested in determining whether these dimensions were independently and uniquely associated with preparatory grief. METHODS: Two hundred advanced cancer patients treated in a Pain Relief and Palliative Care Unit completed the PGAC scale, while researchers recorded data on demographic characteristics, disease status and treatment regimen. RESULTS: The analyses showed that the most significant correlations were found between preparatory grief and age (r = -0.227, p = 0.001), gender (p = 0.006), family status (p = 0.019), performance status (p = 0.010), surgery (p = 0.029), opioids (p = 0.019), and diagnosis (p = 0.038). In the prediction of preparatory grief, the contribution of age, performance status, history of other surgery, gender and opioids is high. CONCLUSIONS: Awareness of the specific patients' demographic and medical characteristics, such as old age, poor performance status, history of other surgery, female gender, and strong opioids, contribute to the prediction of patients' preparatory grief. PMID- 16477679 TI - (1)H relaxation times of metabolites in biological samples obtained with nondestructive ex-vivo slow-MAS NMR. AB - Methods suitable for measuring (1)H relaxation times such as T(1), T(2) and T(1rho) of metabolites in small, intact biological objects including live cells, excised organs and tissues, oil seeds etc. are developed in this work. This was achieved by combining inversion-recovery, spin-echo, or a spin-lock segment with the phase-adjusted spinning sideband (PASS) technique, which was applied at low sample-spinning rates. Here, PASS was used to produce high-resolution (1)H spectra in a nondestructive way so that the relaxation parameters of individual metabolite could be determined. The methodologies were demonstrated by measuring (1)H T(1), T(2), and T(1rho) of metabolites in excised rat liver at a spinning rate of 40 Hz. PMID- 16477680 TI - Order parameters based on (13)C(1)H, (13)C(1)H(2) and (13)C(1)H(3) heteronuclear dipolar powder patterns: a comparison of MAS-based solid-state NMR sequences. AB - Order parameters describing conformational exchange processes on the nanosecond to microsecond timescale can be obtained from powder patterns in solid-state NMR (SSNMR) experiments. Extensions of these experiments to magic-angle spinning (MAS) based high-resolution experiments have been demonstrated, which show a great promise for site-specific probes of biopolymers. In this study, we present a detailed comparison of two pulse sequences, transverse Manfield-Rhim-Elleman Vaughn (T-MREV) and Lee-Goldburg cross-polarization (LGCP), using experimental and simulation tools to explore their utility in the study of order parameters. We discuss systematic errors due to passively coupled (13)C or (1)H nuclei, as well as due to B(1) inhomogeneity. Both pulse sequences can provide quantitative measurements of the order parameter, but the LGCP experiment is capable of greater accuracy provided that the B(1) field is highly homogeneous. The T-MREV experiment is far better compensated for B(1) inhomogeneity, and it also performs better in situations with limited signal. PMID- 16477681 TI - Comparison of the performance of round and rectangular wire in small solenoids for high-field NMR. AB - This paper considers the effects of conductor geometry on the performance of small solenoidal coils for high-field NMR. First, a simple analytical model is presented for investigating the effects of conductor geometry on the current distribution in such coils. The model was used to derive optimum parameters for coils constructed from wire with either rectangular or circular cross-sections as a function of the length-to-diameter ratio. Second, a commercial software package utilizing full three-dimensional finite-element solutions to Maxwell's equations was used to confirm the basic findings of the simple analytical model, and also to compare simulated S/N estimations with experimental NMR spectra acquired with 2.5 mm and 1.0 mm-diameter solenoid coils: reasonable agreement was found. Third, as a demonstration of the usefulness of such coils for mass-limited samples, multidimensional experiments were performed at 750 MHz on approximately 4.7 nmol (41 microg) of PF1061, a protein from Pyrococcus furiosus. PMID- 16477682 TI - Solid-state NMR and computational studies of 4-methyl-2-nitroacetanilide. AB - Studies on the solid-state structure of two polymorphs of 4-methyl-2 nitroacetanilide (MNA) were conducted using magic-angle spinning (13)C, (15)N and (1)H NMR spectroscopy, together with first-principles computations of NMR shielding (including use of a program that takes explicit account of the translational symmetry inherent in crystalline structures). The effects on (13)C chemical shifts of side-chain rotations have been explored. Information derived from these studies was then incorporated within a systematic space-search methodology for elucidation of trial crystallographic structures from powder XRD. PMID- 16477684 TI - Stability constants: comparative study of fitting methods. Determination of second-order complexation constants by (23)Na and (7)Li NMR chemical shift titration. AB - NMR chemical shift titration has been widely used as a method for the determination of stability constants. Systems involving metal-ligand complexation have been investigated using a number of methodologies. There are significant differences in the values reported for stability constants obtained by different experimental methods, such as calorimetry and ion selective electrode (ISE) titrations; nor has NMR chemical shift titration always yielded consistent results. Different researchers have obtained different results for the same system with results differing by as much as an order of magnitude. The chemical shift data are generally plotted against the concentration ratio of the metal and ligand for a set of solutions. A nonlinear least squares fitting method using an analytical solution of the cubic equation for the equilibrium concentration of the free ligand is used in this study and compared with methods used in the literature. Second-order association constants for the LiClO(4):12-crown-4 system in acetonitrile and the NaClO(4):12-crown-4 system in methanol are reported. Formation of both 1:1 and 1:2 metal-ligand complexes are considered. The LiClO(4):12-crown-4 acetonitrile system had been investigated previously by NMR titration but only 1:1 complexation was considered in that study. This study provides convincing evidence that both 1:1 and 1:2 complexes are important, at least, in the lithium system. A Monte Carlo investigation of the propagation of errors from the chemical shifts to the stability constants shows that the choice of data analysis methods may, in part, contribute to discrepancies and that the nonlinear nature of the model can dramatically affect the error limits on the stability constants. PMID- 16477685 TI - High-resolution magic-angle spinning (13)C spectroscopy of brain tissue at natural abundance. AB - High-resolution magic-angle spinning (MAS) (1)H and (13)C magnetic resonance spectroscopy (MRS) has recently been applied to study the metabolism in intact biological tissue samples. Because of the low natural abundance and the low gyromagnetic ratio of the (13)C nuclei, signal enhancement techniques such as cross-polarization (CP) and distortionless enhancement by polarization transfer (DEPT) are often employed in MAS (13)C MRS to improve the detection sensitivity. In this study, several sensitivity enhancement techniques commonly used in liquid and solid-state NMR, including CP, DEPT and nuclear Overhauser enhancement (NOE), were combined with MAS to acquire high-resolution (13)C spectra on intact rat brain tissue at natural abundance, and were compared for their performances. The results showed that different signal enhancement techniques are sensitive to different classes of molecules/metabolites, depending on their molecular weights and mobility. DEPT was found to enhance the signals of low-molecular weight metabolites exclusively, while the signals of lipids, which often are associated with membranes and have relatively lower mobility, were highly sensitive to CP enhancement. PMID- 16477686 TI - Detailed analysis of coupling constants and isotope effects in NMR spectra of isotopomers of (12)C(68) (13)C(2). AB - A preliminary study of the long-range (i.e. two-bond or longer) (13)C--(13)C coupling constants in natural abundance C(70) shows, consistent with recent theoretical calculations by Peralta et al. that the largest long-range J(CC) values for the polar and equatorial sites are clearly smaller than the largest long-range J(CC) values for the other three sites. The unusually large size of the (2)J(CC) couplings between inequivalent carbons in a nonpolar pentagon in C(70) has no analog among (2)J(CC) data reported for planar aromatic compounds. No long-range J(CC) values appear to have been reported for any curved aromatic compounds. In addition, much more precise (1)J(CC) values were obtained for C(70) than was possible about 15 years ago. Comparing the chemical shifts for each of the five isotopomers of C(70) containing only one (13)C nucleus and the frequencies of the satellites for each of the four isotopomers containing two adjacent and inequivalent (13)C nuclei indicates that replacing (12)C with (13)C shields the adjacent (13)C nucleus by 15 to 23 ppb, consistent with the limited (1)Delta(13)C((13/12)C) isotope effect data available on a few small aromatic molecules. Such measurements become possible with natural abundance C(70) only by using a (13)C cryoprobe and a high-field spectrometer (700 MHz). The additional information that could be obtained from a spectrum obtained under ultrahigh resolution conditions is discussed. Secure identification of the singlets arising from the four (12)C(68) (13)C(2) isotopomers with equivalent adjacent (13)C nuclei is necessary to allow the largest long-range J(CC) values to be precisely determined. The presence of numerous isotopomers containing two or more (13)C nuclei would present a great challenge in interpreting the various signals in a spectrum obtained under ultrahigh resolution conditions. PMID- 16477687 TI - On the use of pseudocontact shifts in the structure determination of metalloproteins. AB - The utility of pseudocontact shifts in the structure refinement of metalloproteins has been evaluated using a native, paramagnetic Cu(2+) metalloprotein, plastocyanin from Anabaena variabilis (A.v.), as a model protein. First, the possibility of detecting signals of nuclei spatially close to the paramagnetic metal ion is investigated using the WEFT pulse sequence in combination with the conventional TOCSY and (1)H-(15)N HSQC sequences. Second, the importance of the electrical charge of the metal ion for the determination of correct pseudocontact shifts from the obtained chemical shifts is evaluated. Thus, using both the Cu(+) plastocyanin and Cd(2+)-substituted plastocyanin as the diamagnetic references, it is found that the Cd(2+)-substituted protein with the same electrical charge of the metal ion as the paramagnetic Cu(2+) plastocyanin provides the most appropriate diamagnetic reference signals. Third, it is found that reliable pseudocontact shifts cannot be obtained from the chemical shifts of the (15)N nuclei in plastocyanin, most likely because these shifts are highly dependent on even minor differences in the structure of the paramagnetic and diamagnetic proteins. Finally, the quality of the obtained (1)H pseudocontact shifts, as well as the possibility of improving the accuracy of the obtained structure, is demonstrated by incorporating the shifts as restraints in a refinement of the solution structure of A.v. plastocyanin. It is found that incorporation of the pseudocontact shifts enhances the precision of the structure in regions with only few NOE restraints and improves the accuracy of the overall structure. PMID- 16477688 TI - Phase transition of L-Ser monohydrate crystal studied by (13)C solid-state NMR. AB - We used gravimetric analysis (GA) and (13)C solid-state nuclear magnetic resonance (NMR) to study solid-phase transition from the transparent single crystal of L-serine (L-Ser) monohydrate to a turbid powder. We found that L-Ser monohydrate loses water molecules and transforms into an anhydrate, thus experimentally demonstrating Frey's assumption. Application of a handmade cross polarization (CP) NMR probe with a saddle-type coil to the oriented crystal of the L-Ser monohydrate revealed the dehydration mechanism. Furthermore, the chemical shift tensor components of the carboxyl carbon in L-Ser monohydrate were determined. The difference in the tensor component of delta(22) between the monohydrate and anhydrate forms was more than 7 ppm, probably owing to differences in the hydrogen-bonding structure of each form. PMID- 16477689 TI - Solid-state (14)N MAS NMR of ammonium ions as a spy to structural insights for ammonium salts. AB - The high resolution offered by magic-angle spinning (MAS), when compared to the static condition in solid-state NMR of powders, has been used to full advantage in a (14)N MAS NMR study of some ammonium salts: CH(3)NH(3)Cl, (NH(4))(2)(COO)(2) x H(2)O, (CH(3))(3)(C(6)H(5)CH(2))NCl, (CH(3))(3)(C(6)H(5))NI, [(n C(4)H(9))(4)N](2)Mo(2)O(7), (NH(4))(2)HPO(4), and NH(4)H(2)PO(4). It is shown that the high-quality (14)N MAS NMR spectra, which can be obtained for these salts, allow determination of the (14)N quadrupole coupling parameters, i.e. C(Q) (the quadrupole coupling constant) and eta(Q) (the asymmetry parameter), with very high precision. In particular, it is shown that precise C(Q), eta(Q) parameters can be determined for at least two different (14)N sites in case the individual spinning-sideband (ssb) intensities arise from a single manifold of ssbs, i.e. the ssbs for the two sites cannot be resolved. This feature of (14)N MAS NMR, which is the first demonstration for manifolds of ssb in MAS NMR without the potential information from a central transition, becomes especially useful at the slow spinning frequencies (nu(r) = 1000-1500 Hz) applied to some of the ammonium salts studied here. The detection of the number of sites has been confirmed by the corresponding crystal structures determined from single-crystal X-ray diffraction (XRD), either in this work for the unknown structure of benzyl trimethylammonium chloride or from reports in the literature. The magnitudes of the (14)N quadrupole coupling constants for the ammonium salts studied here are in the range from C(Q) approximately 20 kHz to 1 MHz while the asymmetry parameters span the full range 0 < or = eta(Q) < or = 1. Clearly, the (14)N quadrupole coupling parameters (C(Q), eta(Q)) for ammonium ions appear highly sensitive toward crystal structure and therefore appreciably more informative for the characterization of ammonium salts in comparison to the isotropic (14)N (or (15)N) chemical shifts. PMID- 16477690 TI - Solid-state (207)Pb NMR studies of lead-group 16 and mixed transition-metal/lead group 16 element-containing materials. AB - (207)Pb solid-state NMR studies have been conducted on binary lead-group 16 and mixed transition-metal/lead group 16 materials, correlating the NMR chemical shifts of the materials with their structures. The experimental results show that the (207)Pb chemical shifts are strongly influenced by the local electronic structure. Data are reported for lead selenide, lead selenate, calcium plumbate, strontium plumbite, barium plumbite, lead borate, lead zirconate, lead tungstate, lead meta-tantalate, lead niobate, lead molybdate, lead meta-vanadate, lead sulfite, and lead sulfate. PMID- 16477691 TI - Solid-state NMR investigation of sodium nucleotide complexes. AB - Solid-state NMR has been used to analyze the chemical environments of sodium sites in powdered crystalline samples of sodium nucleotide complexes. Three of the studied complexes have been previously characterized structurally by crystallography (disodium deoxycytidine-5'-monophosphate heptahydrate, disodium deoxyuridine-5'-monophosphate pentahydrate and disodium adensoine-5'-triphosphate trihydrate). For these salts, the nuclear quadrupole coupling parameters measured by (23)Na multiple-quantum magic-angle-spinning NMR could be readily correlated with sodium ion coordination environments. Furthermore, two complexes that had not been previously characterized structurally, disodium uridine-3'-monophosphate and a disodium uridine-3'-monophosphate/disodium uridine-2'-monophosphate mix, were identified by solid-state NMR. A spectroscopic assignment of the four sites of an additional salt, disodium adensoine-5'-triphosphate trihydrate, is also presented and discussed within the context of creating a general approach for the spectroscopic assignment of multiple sites in sodium nucleotide complexes. PMID- 16477692 TI - Interactions between pairs of antimalarial drugs studied by experimental and ab initio (13)C NMR chemical shifts. AB - The interactions between the antimalarial drugs chloroquine (CQ) and amodiaquine (AQ), chloroquine and quinine (QN), and amodiaquine and quinine are studied by (13)C NMR. Experimental changes in chemical shift are compared to nucleus independent chemical shifts to determine the best structure of the complex formed by each drug pair in solution. Structures of the CQ-AQ and CQ-QN complexes are found to be similar to those found previously for the drug dimers. On the other hand, the best solution structure for the AQ-QN complex suggests that the quinoline rings of the two drugs are at an angle with respect to each other. PMID- 16477693 TI - Molecular reorientation of CD(4) in gas-phase mixtures. AB - Spin-lattice relaxation times were measured for the deuterons in CD(4) in pure gas and in mixtures with the following buffer gases: Ar, Kr, Xe, HCl, N(2), CO, CO(2), CF(4), and SF(6). Effective collision cross sections sigma(theta, 2) for the molecular reorientation of CD(4) in collisions with these ten molecules are obtained as a function of temperature. These cross sections are compared with the corresponding cross sections sigma(J) obtained from (1)H spin-rotation relaxation in mixtures of CH(4) with the same set of buffer gases. Various classical reorientation models typically applied in liquids predict different ratios of the reduced correlation times for the reorientation of spherical tops. The Langevin model comes closest to predicting the magnitude of the sigma(theta, 2)/sigma(J) ratio that we obtain for CD(4). PMID- 16477694 TI - Chemical shift imaging (CSI) by precise object displacement. AB - A mechanical device (NMR lift) has been built to displace vertically an object (typically an NMR sample tube) inside the NMR probe with an accuracy of 1 microm. A series of single pulse experiments are performed for incremented vertical positions of the sample. With a sufficiently spatially selective radio-frequency (r.f.) field, one obtains chemical shift information along the displacement direction (one-dimensional chemical shift imaging (CSI)). Knowing the vertical r.f. field profile (the amplitude of the r.f. field along the vertical direction), one can reconstruct the spectrum associated with all the slices corresponding to consecutive sample positions and improve the spatial resolution, which is simply related to the accuracy of the displacement device. Beside tests performed on phantoms, the method has been applied to solvent penetration in polymers and to benzene diffusion in a heterogeneous zeolite medium. PMID- 16477695 TI - Conformational analyses of 2,3-dihydroxypropanoic acid as a function of solvent and ionization state as determined by NMR spectroscopy. AB - Vicinal (1)H--(1)H coupling constants were used to determine the conformational preferences of 2,3-dihydroxypropanoic acid (1) (DL-glyceric acid) in various solvents and its different carboxyl ionization states. The stereospecific assignments of J(12) and J(13) were confirmed through the point-group substitution of the C-3 hydrogen with deuterium, yielding rac-(2SR,3RS)-[3-(2)H] 1, and the observation of only J(13) in the (1)H NMR spectra. While hydrogen bonding and steric strain may be expected to drive the conformational equilibrium, their role is overshadowed by a profound gauche effect between the vicinal hydroxyl groups that mimics other substituted ethanes, such as 1,2 ethanediol and 1,2-difluoroethane. At low pH, the conformational equilibrium is heavily weighted toward the gauche-hydroxyl rotamers with a range of 81% in DMSO d(6) to 92% in tert-butyl alcohol-d(10). At high pH, the equilibrium exhibits a larger dependence upon the polarity and solvating capability of the medium, although the gauche effect still dominates in D(2)O, 1,4-dioxane-d(8), methanol d(4), and ethanol-d(6) (96, 89, 85, and 83% gauche-hydroxyls respectively). The observed preference for the gauche-hydroxyl rotamers is believed to stem primarily from hyperconjugative sigma(C--H) --> sigma*(C--OH) interactions. PMID- 16477696 TI - Effect of rapidly relaxed electron spin anisotropically coupled to nearby nuclear partners. AB - In practice, many situations arise when a perturbed nuclear spin relies upon the aid of a rapidly relaxed spin neighbor in order to realize thermal equilibrium. Conventional treatments view the efficiently relaxed spin as part of the 'lattice', invoke the secular approximation, and consider the associated time correlation of the lattice variables, phenomenologically. Recently, an ab initio perturbative approach has been proposed for investigation of these spin systems. In this work, a similar formalism is applied to the scenario, in which nuclear spin relaxation is effected via an anisotropically coupled, efficiently relaxed, spin. Interesting conflicts with standard theory are revealed. Furthermore, although left mostly unexplored, this approach lends insight into numerous related aspects of magnetic relaxation including separation of timescales, distinction between spin and spatial averaging, paramagnetic relaxation, Curie spin relaxation, and the dynamic frequency shift. PMID- 16477697 TI - A comparative theoretical study of dipeptide solvation in water. AB - Molecular dynamics studies have been performed on the zwitterionic form of the dipeptide glycine-alanine in water, with focus on the solvation and electrostatic properties using a range of theoretical methods, from purely classical force fields, through mixed quantum mechanical/molecular mechanical simulations, to fully quantum mechanical Car-Parrinello calculations. The results of these studies show that the solvation pattern is similar for all methods used for most atoms in the dipeptide, but can differ substantially for some groups; namely the carboxy and aminoterminii, and the backbone amid NH group. This might have implications in other theoretical studies of peptides and proteins with charged NH(3) (+) and -CO(2) (-) side chains solvated in water. Hybrid quantum mechanical/molecular mechanical simulations successfully reproduce the solvation patterns from the fully quantum mechanical simulations (PACS numbers: 87.14.Ee, 87.15.Aa, 87.15.He, 71.15.Pd). PMID- 16477699 TI - Bulgarian hospital admits role in illegal transplants. PMID- 16477698 TI - Reply to: 'The predictive value of the sperm chromatin structure assay (SCSA)'--a response from the SCSA inventor. PMID- 16477700 TI - Canadian petrochemical plants blamed for gender imbalance. PMID- 16477701 TI - Spaniards sceptical of new smoking ban. PMID- 16477702 TI - Private sector's administration costs rival Medicare. PMID- 16477703 TI - Let's talk about sex. PMID- 16477704 TI - Round-up from the 12th annual Retrovirus Conference. Highlights of news items from the Retrovirus Conference in Boston this past February. PMID- 16477705 TI - 12th Retrovirus Conference. Better treatments and drugs in the pipeline. Promising research continues to move forward. PMID- 16477706 TI - 12th Retrovirus Conference. Super bug or super dud? CROI forum provides particulars of the New York case. PMID- 16477707 TI - 12th Retrovirus Conference. Viramune to prevent mother-to-child transmission. Moving from to consensus. PMID- 16477708 TI - It's the best time to get HIV, and the worst. While the virus finds its way around the drugs, newer treatments hold hope. PMID- 16477709 TI - Sexually-transmitted infections for HIV-positive people. Signs and symptoms of common STDs, warts and all. PMID- 16477710 TI - Surviving abuse--and taking care of yourself. A therapist gives strategies for healing. PMID- 16477712 TI - The phoenix rises again. 'One man's story' continues a year after his liver transplant. PMID- 16477711 TI - Sex and prevention. Learning from a group of men of color. PMID- 16477713 TI - Evidence that cyclic GMP regulates myosin interaction with the cytoskeleton during chemotaxis of Dictyostelium. AB - Amoebae of Dictyostelium discoideum respond to a chemotactic cyclic AMP stimulus within 10 s by the formation of an intracellular peak of cyclic GMP. In wild-type cells the cyclic GMP is rapidly degraded by a cyclic GMP-specific phosphodiesterase. In "streamer F" mutants this enzyme structural gene, and the cyclic GMP persists several times longer than the normal period, an effect that is correlated with a persistence in cell elongation during the chemotactic movement phase. In this study we have used the streamer mutants NP368 and NP377 and their parental strain XP55, to study changes in cytoskeletal proteins during the chemotactic response. We have studied three proteins that change their association with the cytoskeleton after stimulation of amoebae with the chemoattractant cyclic AMP: (1) actin, (2) a protein with an apparent Mr of 190 x 10(3) and (3) myosin heavy chain. Both actin and the 190 x 10(3) Mr protein were found to accumulate rapidly int he cytoskeleton after cyclic AMP stimulation, with a sharp peak at 5 s, and showed similar changes in the parental and streamer mutants. However, the cytoskeletal level of myosin heavy chain showed different pattern of changes, which also compared with the parental strain XP55. In XP55 myosin heavy chain showed an initial drop after cyclic AMP stimulation, with a trough at 3-10 s followed by a rapid rise to a sharp peak at 20-25 s. In contrast, the myosin heavy chain in the streamer mutants produced a broad peak that persisted several times longer than the parental strain. We conclude that in the streamer mutants the defect in cyclic GMP phosphodiesterase that produces the broad peak of cyclic GMP is casually correlated with the broad peak of cytoskeletal myosin, and we suggest that this is connected with the observed phenotype of prolonged cell elongation during chemotaxis in these mutants. PMID- 16477714 TI - Women's Medical Professional Corporation v. Taft. PMID- 16477715 TI - [The response to emergency. Epistaxis--hemostatic techniques]. PMID- 16477716 TI - Planned Parenthood of Southern Arizona v. Lawall. PMID- 16477717 TI - Lake v. Arnold. PMID- 16477718 TI - Women's Medical Center of Northwestern Houston v. Bell. PMID- 16477719 TI - [The response to emergency. Foreign bodies in respiratory system and esophagus]. PMID- 16477720 TI - Planned Parenthood of Southern Arizona v. Lawall. PMID- 16477722 TI - Oregon v. Ashcroft. PMID- 16477721 TI - Planned Parenthood of Rocky Mountains v. Owens. PMID- 16477723 TI - Britell v. United States. PMID- 16477725 TI - Northwestern Memorial Hospital v. Ashcroft. PMID- 16477724 TI - Planned Parenthood of Minnesota/South Dakota v. Rounds. PMID- 16477726 TI - Tucson Woman's Clinic v. Eden. PMID- 16477727 TI - Editorial note: Diversity and Bakke - an unfortunate and unnecessary consequence. PMID- 16477728 TI - Proceedings of the XXVII Annual Congress of the Spanish Society for Paediatric Endocrinology (SEEP). May 5-7, 2005. Oviedo, Spain. PMID- 16477729 TI - Vicarious liability of the employer of an apparent servant. PMID- 16477730 TI - [Trends in change of names for mental diseases in japan]. PMID- 16477731 TI - [Is lethal catatonia a non-drug induced malignant syndrome?]. PMID- 16477732 TI - In re Estate of Kolacy. PMID- 16477733 TI - Planned Parenthood of Central New Jersey v. Farmer. PMID- 16477734 TI - Health Insurance Portability and Accountability Act of 1996. Public Law 104-191. PMID- 16477735 TI - In re Duran. PMID- 16477737 TI - [The ideal in child psychiatry]. PMID- 16477736 TI - [International support from psychiatrists for Tsunami disaster in Asian developing countries]. PMID- 16477738 TI - [Response of the Japanese Health and Welfare Agency to the educational system for the specialty certification of child psychiatrists]. PMID- 16477739 TI - [Views of a child psychiatrist engaging in child counseling in Japan]. PMID- 16477740 TI - [Views of a child-adolescent psychiatrist in Japan]. PMID- 16477741 TI - [Views of a general psychiatrist on child-adolescent psychology in japan]. PMID- 16477742 TI - [Views of pediatric physicians on child mental health and child psychiatry in Japan]. PMID- 16477743 TI - [Why group therapy?]. PMID- 16477745 TI - Performance anxiety. PMID- 16477744 TI - [Developmental disorders viewed from child and adolescent psychiatry]. PMID- 16477746 TI - Drug-error detectives. PMID- 16477747 TI - Watching their backs. PMID- 16477748 TI - Fast-track lifesaving. PMID- 16477749 TI - Meeting of the minds. PMID- 16477750 TI - Navigating the quality waters. PMID- 16477751 TI - The bell curve. PMID- 16477752 TI - Deadly & deceptive. PMID- 16477753 TI - Lost in translation. PMID- 16477754 TI - From informed consent to informed coercion? PMID- 16477755 TI - Our obligation to report our performance to the public. PMID- 16477756 TI - Report cards could damage quality of care. PMID- 16477757 TI - Rupturing ascending aorta missed in the emergency room. Failure to properly evaluate chest pain in patients who subsequently die. AB - This article reports on 4 patients who were not properly evaluated in the emergency room when they presented with acute chest pain. All were discharged and eventually died from a ruptured ascending aorta. It calls attention to a serious problem that when mishandled can lead to a fatal outcome. PMID- 16477758 TI - Implementing evidence-based practices to improve quality. One medical group's experience. AB - HealthPartners Medical Group and Clinics uses evidence-based guidelines for diabetes and other chronic diseases. This article reviews how HealthPartners has redesigned its care processes and implements the diabetes guidelines. It also summarizes the lessons the organization has learned about how to choose guidelines and how to get the most from them. PMID- 16477759 TI - Adopting an electronic health record. Keys to success. AB - Stratis Health, through its capacity as Minnesota's Medicare Quality Improvement Organization, is working to promote successful implementation of electronic health records (EHRs) in primary care clinics in the state. This article focuses on lessons learned while helping primary care clinics adopt EHRs. It also includes observations on the current status of EHR adoption in Minnesota clinics. Sixty-eight percent of 341 clinics responding to a survey have implemented, are in the process of implementing, or plan to implement electronic health record systems during the next 2 years. PMID- 16477760 TI - The case of Mr. Bristow. PMID- 16477761 TI - Plasma folate levels in men with type 2 diabetes. AB - Limited data suggest that folate levels are higher in patients with type 2 diabetes than in subjects with normal glucose tolerance (NGT). We compared the fasting plasma folate, glucose (FPG), body mass index (BMI), and supplementary vitamin use among male subjects with NGT, those with impaired glucose tolerance (IGT), those with newly diagnosed type 2 diabetes, and those with previously diagnosed type 2 diabetes. Plasma folate of patients with newly diagnosed diabetes and that of patients with previously diagnosed diabetes was significantly higher than that of NGT subjects (p < 0.001). Prevalence of vitamin use was lower in newly diagnosed or previously diagnosed diabetic patients compared with non-diabetic subjects. Self-rated vegetable intake was similar among the four groups. FPG, BMI, triglycerides, and systolic blood pressure correlated with plasma folate levels independently of lifestyle factors studied. These results suggest that plasma folate levels are elevated in male diabetic patients independently of health-conscious behavior that is recommended for diabetic people. PMID- 16477762 TI - Nutrient intake adequacy in schoolchildren from a Mediterranean area (southern Spain). Influence of the use of the school canteen. AB - The aim of the present study was to compare dietary adequacy in 521 schoolchildren in Granada (Spain) with national recommendations and to determine possible differences regarding gender or the use of the school canteen. A food record was used for four consecutive days to evaluate the intake of nutrients and foods. With regard to boys' and girls' intake of the main groups of foods, we observed a tendency for the boys to consume greater quantities of dairy products, cereals, potatoes, and fruit, although the differences were not statistically significant. The children who lunched at school consumed fish, eggs, dairy products, vegetables, and cereals more frequently than those who did so at home. The mean daily intake of energy and nutrients indicates that the studied population exceeded the recommended limits in the proportion of energy derived from protein and fat, while carbohydrate intake was slightly deficient. The children who lunched at school showed a more favorable profile for energy and nutrient intake, with a significantly higher intake of energy, carbohydrates, total fat, polyunsaturated fatty acids (PUFA), monounsaturated fatty acids (MUFA), vitamins B, and E, and calcium, with respect to the children who had lunch at home. PMID- 16477763 TI - Effect of B1-, B6- and iron intake during pregnancy on neonatal behavior. AB - The aim of this study was to analyze how micronutrient intake during preconception and pregnancy affects neonatal behavior. A total of 66 healthy women volunteers were studied during preconception and in weeks 6, 10, 26, and 38 of pregnancy using the seven-day dietary record. The behavior of the newborn infant was assessed after three days of life using the Neonatal Behavioral Assessment Scale (NBAS). Multiple linear regression models were built and adjusted for the confounding variables in each of the periods studied and for each one of the various NBAS clusters. The intake of vitamins B1 and B6 in the sixth week of pregnancy and of iron in the 38th week of pregnancy have a positive and significant effect on the motor cluster of the NBAS independently of the body mass index (BMI), preconception age, gestational age, educational level, whether the mother is a smoker, the mother's personality dimensions, and the weight and sex of the newborn. The intake of B1, B6, and iron during pregnancy might contribute to the neuromotor maturity of the newborn. PMID- 16477764 TI - The effect of vitamin E supplementation on serum DHEA and neopterin levels in elderly subjects. AB - Contradictory results have been published on the immune-stimulating effects of vitamin E. Using a randomized placebo-controlled design, the effect of 15 month's daily supplementation with 200 mg vitamin E on two biomarkers of immunocompetence, i.e. serum DHEA sulfate ester (DHEA-S) and neopterin, was studied. Of the 100 relatively healthy Dutch elderly subjects included in the study, 50 were supplemented with vitamin E and 50 received placebo. As compared to placebo, vitamin E supplementation affected neither serum DHEA-S nor serum neopterin level. This corroborates with the finding that vitamin E supplementation did not affect infection-related severity measures, i.e. total number of days with respiratory infection, and total duration of the infections. It is concluded that vitamin E supplementation does not substantially alter the immunocompetence markers DHEA and neopterin in elderly subjects, and may explain our recently reported failure of vitamin E supplementation to afford protection against acute respiratory infections. PMID- 16477765 TI - Oxidative stress and antioxidant status in rat blood, liver and muscle: effect of dietary lipid, carnitine and exercise. AB - The purpose of this study was to determine the effect of dietary fat, carnitine supplementation, and exercise on oxidative damage and antioxidant status. Male Wistar rats (60 days old) were fed diets containing either hydrogenated fat (HF) or peanut oil (PO) with or without 0.5 mg % (of dry diet) carnitine. The rats were given exercise, i.e. swimming for 60 minutes, for 6 days/week for 6 months under each dietary condition. The blood malondialdehyde (MDA) level was higher in PO-fed rats, more so in exercising ones, while the same was not altered in carnitine-supplemented rats irrespective of the dietary fat or physical activity. The MDA level was significantly decreased in muscle, while increased in liver, of carnitine-fed rats. The blood glutathione (GSH) level also significantly increased in exercising rats as compared to sedentary ones, while carnitine supplementation elevated it in all the groups. Exercise and carnitine supplementation significantly lowered GSH levels in liver while increasing it in muscle. The glutathione peroxidase (GPX) activity was significantly increased in blood and muscle from PO-fed exercising rats as compared to sedentary ones, while carnitine supplementation elevated GPX activity in all the groups. The liver and muscle catalase (CAT) activities were significantly increased in PO-fed exercising rats, while carnitine did not have any effect. The pro-oxidative effect of the monounsaturated fatty acid (MUFA)-rich PO diet and prolonged regular exercise was less pronounced due to augmented antioxidant enzymes, GPX and CAT, induced by training to protect against the oxidative stress, while carnitine supplementation could help to counter lipid peroxidation due to exercise through redistribution of GSH from liver to blood and muscle. PMID- 16477766 TI - Long term effects of oral vitamin E supplement in type II diabetic patients. AB - This triple-blind, placebo-controlled clinical trial was conducted to determine the effect of the vitamin E on fasting blood sugar (FBS), serum insulin, and glycated hemoglobin (GHb) in type 11 diabetic patients (NIDDM). A total of 100 patients, with no complications, aged 20-60 years old were chosen from those consulting the Isfahan Social Security Service Diabetes Clinic and divided randomly into two treated and placebo groups, and matched for age, sex, level of education, and occupation. The treated and placebo groups were given vitamin E tablets (200 IU/day) and placebo respectively. Serum vitamin E, total cholesterol (TC), triglycerides (TG), FBS, insulin, and GHb were measured at the beginning and at the end of the study (a period of 27 weeks); FBS, GHb and insulin levels were also determined several times during the period. Blood lipids and FBS were measured using the ELAN 2000 autoanalyzer at the Isfahan Cardiovascular Research Center, while for measuring insulin the enzyme-linked immunosorbent assay (ELISA) method was used; GHb was determined calorimetrically (thiobarbituric acid), and for vitamin E measurements the Hansen and Warwick method was used, by which the vitamin E was determined fluorometrically. The findings of this study show no effect of vitamin E supplementation in the patients: GHb did not change appreciably, FBS was reduced nonsignificantly (-4.3% in the treated group vs. 14.0% in the placebo group, p < 0.05). In the case of insulin, no increase was seen; instead, a decrease was observed (slightly more than 17% in the two groups, p = 0.15). No changes were observed in the levels of blood lipids. It was concluded that a daily vitamin E supplement of 200 IU for a period of 27 weeks does not affect insulin, GHb, or FBS in type II diabetic patients. However, since this antioxidant vitamin is beneficial in other ways in these patients, it would seem justified to recommend its use. Certainly, more extensive research is necessary to draw definite conclusions. PMID- 16477767 TI - Evaluation of waist circumference to predict cardiova scular risk factors in an overweight Tehranian population: findings from Tehran Lipid and Glucose Study. AB - This study was conducted to evaluate the waist circumference (WC) cut-off points to predict cardiovascular risk factors in the overweight Tehranian population. Anthropometric measures, blood pressure, and biochemical analyses were evaluated for the 15,005 participants of the Tehran Lipid and Glucose Study. Three thousand sixty-five subjects aged 18-74 years with a body mass index of 25-29.9 were enrolled in this study. Abdominal obesity was defined as WC > or =102 cm for men and > or =88 cm for women. Sensitivity of WC > or =102 cm to detect various cardiovascular risk factors for men aged 35-54 years was between 5% and 14%, and for men aged 55-74 years, was between 12% and 19%. The specificity of this cut off point was between 93% and 98% and between 86% and 96% for corresponding age categories, respectively. WC > or =88 cm had a sensitivity of between 28% and 41 % for identifying cardiovascular risk factors in women aged 18-34 years. Sensitivity tended to increase with age and specificity tended to decrease with age in both genders. These cut-off points had the highest positive predictive value for the more prevalent risk factors in both genders. The negative predictive values were different for various risk factors among age groups. The classic cut-off points of WC failed to provide adequate evidence for the use of WC in detecting cardiovascular risk factors. Further studies should be conducted to determine optimal WC cut-off points for Iranians. PMID- 16477768 TI - Effects of increased dietary protein and energy on composition and functional capacities of blood mononuclear cells from vaccinated, neonatal calves. AB - Effects of increased protein and energy provided by an intensified milk replacer on the antigen-specific, cell-mediated immune response of the neonatal calf were examined. Calves were fed a standard (0.45 kg/day of a 20% crude protein, 20% fat milk replacer; n=11) or intensified (1.14 kg/day of a 28% crude protein, 20% fat milk replacer; n=11) diet from 0 to 6 weeks of age. All calves were vaccinated with Mycobacterium bovis bacillus Calmette-Guerin (BCG) at 1 week of age. The daily weight gain of intensified-diet calves (0.62 kg/day) was greater than the weight gain of standard-diet calves (0.29 kg/day). Liver, kidney, heart, thymus, and subcervical lymph nodes from intensified-diet calves were heavier than the same organs from standard-diet calves. Flow cytometric analysis of peripheral blood mononuclear cell (PBMC) populations indicated that CD4+ cells, gamma delta TCR+ cells, and monocyte percentages, although unaffected by diet during the first 5 weeks of the study, were higher in intensified-diet calves at week 6. The decline in gamma delta TCR+ cell percentages and increase in B cell percentages with increasing age seen in all calves are characteristic of the maturing immune system of the calf. CD8+ T cell or B cell percentages were not affected by diet. In intensified-diet calves, percentages of CD4+ expressing interleukin-2 receptor increased and percentages of gamma delta TCR+ cells expressing interleukin-2 receptor decreased with time. The same populations in standard-diet calves did not change with time. Percentages of CD4+ and CD8+ T cells, and B cells expressing MHC class II antigen, were unaffected by diet or age. Although mitogen induced interferon (IFN)-gamma and nitric oxide (NO) secretion increased with age for all calves, PBMC from intensified-diet calves produced less IFN-gamma and more NO than did cells from standard-diet calves at week 6 of the study. Antigen induced secretion of IFN-gamma and NO also increased with age but was unaffected by diet. Antigen-elicited delayed-type hypersensitivity was unaffected by diet, suggesting increased dietary protein and energy did not alter adaptive immunity in vivo. Overall, these results suggest that feeding calves a commercially available, intensified milk replacer affects minimally the composition and functional capacities of PBMC populations. Additional research is necessary to determine whether these subtle effects influence the calf's susceptibility to infectious disease. PMID- 16477769 TI - Case-based teaching and learning. PMID- 16477770 TI - Hospital protocols and evidence-based therapies: the importance of integrating aldosterone blockade into the management of patients with post-acute myocardial infarction heart failure. AB - Left ventricular systolic dysfunction (LVSD) and clinical heart failure are common complications of acute myocardial infarction (AMI) and result in substantially increased mortality and morbidity. Evidence-based cardiovascular protective therapies, including angiotensin-converting enzyme inhibitors, beta blockers, antiplatelet agents, and lipid-lowering medications, improve outcomes for these patients. However, this population is significantly undertreated with these guideline-recommended agents. Critical pathways have been demonstrated to improve the quality and consistency of treatment; as such, the new American College of Cardiology/American Heart Association (ACC/AHA) guidelines for the management of patients with ST-elevation myocardial infarction (STEMI) recommend that critical pathways be implemented for the management of these patients. The recent Eplerenone Post-acute Myocardial Infarction Heart Failure Efficacy and Survival Study (EPHESUS) demonstrates that eplerenone, a selective aldosterone blocker, has incremental benefit in decreasing mortality and morbidity when used with standard care therapies in patients post AMI with heart failure and LVSD. The clinical trial evidence coupled with the national guidelines provides a strong rationale for routine incorporation of aldosterone blockade into new or already established critical pathways for AMI complicated by LVSD and heart failure. PMID- 16477771 TI - Left ventricular apical ballooning: not an uncommon variant of acute myocardial infarction in women. AB - BACKGROUND: Left ventricular apical ballooning, a new syndrome recently described in Japan, is characterized by chest pain, electrocardiographic changes mimicking acute myocardial infarction, and transient apical dyskinesia with normal coronary arteries. Although several studies have defined the clinical characteristics, the prevalence of this syndrome remains unclear. HYPOTHESIS: This study sought to determine the prevalence of left ventricular apical ballooning syndrome. METHODS: From January 2002 to September 2004, clinical, echocardiographic, and angiographic data of hospitalization and follow-up were collected from 638 consecutive patients referred to our Heart Institute for primary percutaneous intervention. RESULTS: Thirteen patients (2%) were diagnosed with transient left ventricular apical ballooning. All but one patient were women, representing a 6% incidence for the female patients with acute myocardial infarction. A triggering factor was identified in eight. One patient died of cardiogenic shock. Left ventricular systolic function recovered completely within 4-5 weeks in the remaining 12 survivors. CONCLUSION: This syndrome is not uncommon and should be considered particularly in female patients presenting with acute myocardial infarction. PMID- 16477772 TI - Cardiac sequelae in Brooklyn after the September 11 terrorist attacks. AB - BACKGROUND: Terrorism, such as the attacks on the World Trade Center (WTC) on September 11, 2001, result in higher stress-related disorders, especially in those persons in close proximity. HYPOTHESIS: Cardiac events resulting from the September 11, 2001 tragedy have not been detailed near the WTC. METHODS: Patients admitted to the Telemetry and Coronary Care Units at New York Methodist Hospital 4 miles from the WTC 60 days prior to and after the September 11 attacks were analyzed. In all, 1,653 admissions were reviewed: 427 records pre 9/11/2001 and 422, 393, and 411 records in the post-9/11/2001, pre-9/11/2000, and post 9/11/2000 data sets, respectively. Patients were categorized based on diagnosis: acute myocardial infarction (MI), unstable angina (UA), tachyarrhythmia (TA), and others (including syncope and congestive heart failure). RESULTS: There was a significant difference in the proportion of the various cardiac diagnoses post 9/11/01 (p = 0.008 by chi-square analysis). Compared with pre 9/11/2001, there were significantly more patients with acute MI (15.5 vs. 11.2%) and TA (19.9 vs. 13.6%) but fewer with UA (39.6 vs. 47.3%) after the terrorist attacks. The distribution of cardiac events during a similar period of time in 2000 revealed no such pattern. CONCLUSIONS: There was a significant increase in acute MI and TA and a smaller increase in UA after the September 11 attacks. The difference did not appear to be due to temporal variation. It appears that stress likely contributed to an increase in TA by itself or with ischemia, resulting in progression of UA to acute MI. PMID- 16477773 TI - Effectiveness of nesiritide on dialysis or all-cause mortality in patients undergoing cardiothoracic surgery. AB - BACKGROUND: Natriuretic peptides have been shown to have favorable renal effects. However, recent evidence suggests potential renal side effects in patients with congestive heart failure. HYPOTHESIS: This study examined the effect of nesiritide (human B-type natriuretic peptide) on hemodialysis or death in patients undergoing cardiothoracic surgery. METHODS: This retrospective cohort study included patients (n = 940) undergoing nontransplant adult cardiothoracic surgery between July 2001 and February 2004. Patients receiving nesiritide within 3 days after and not before surgery (n = 151) were compared with those not given nesiritide (n = 789) for incidence of hemodialysis or in-hospital death by Day 21 (HD/death). Patients with preexisting dialysis and intraoperative deaths were excluded. Forward inclusion multiple logistic regression was used based on published risk factors for HD/death. RESULTS: Of 940 patients (318 coronary artery bypass graft, 348 valve, and 274 thoracic aorta), 36 required dialysis and 60 patients died (HD/death; n = 77). Adjusted for significant confounders (gender, age, procedure, intra-aortic balloon, baseline serum creatinine mg/dl [SCr], 1 day % SCr increase), nesiritide showed a statistically nonsignificant HD/death reduction (odds ratio [OR], 0.58; 95% confidence interval [CI], 0.29 1.17; p = 0.129) in the group as a whole. When stratified by baseline SCr, a significant benefit was noted in patients with SCr > 1.0 (OR, 0.35; 95% CI 0.14 0.87; p = 0.024), while no significant effect was found in patients with SCr < 1.0 (OR, 1.55; 95% CI 0.48-5.07, p = 0.465). CONCLUSIONS: Nesiritide appears promising in reducing the risk of dialysis or death in patients with SCr > 1.0 undergoing cardiothoracic surgery; however, no effect was noted with SCr < 1.0. This study provides strong rationale for a randomized trial. PMID- 16477774 TI - Relationship between true fasting plasma insulin level and angiographic characteristics of coronary atherosclerosis. AB - BACKGROUND: Reports about the relationships between specific insulin concentration and coronary heart disease risk are controversial. HYPOTHESIS: The objective of this study was to examine the association between insulin level and the severity of coronary atherosclerosis. METHODS: The study population consisted of 507 consecutive patients (376 men and 131 women) who underwent coronary angiography for suspected or known coronary atherosclerosis. The patients' habits of smoking and drinking were investigated, and anthropometric measurements including body mass index, systolic and diastolic blood pressures, as well as plasma measurements including lipids and blood glucose were taken. The true insulin level was measured using a highly sensitive two-site sandwich ELISA. The severity of coronary atherosclerosis was defined by the Gensini score system. The statistical methods including Kruskal-Wallis test, chi-square analysis, Spearman correlation analysis, and multivariate stepwise linear regression analysis were employed to explore the relationship between specific insulin level and coronary atherosclerosis. RESULTS: When the Gensini score was examined as a categorical variable classified by tertile values, subjects with a high Gensini score had significantly higher values of fasting plasma specific insulin level (p = 0.022). The Spearman correlation analysis suggest that the Gensini score correlated significantly with true insulin (mIU/l) (r = 0.095, p = 0.033). However, the results from the multivariate stepwise linear regression analysis show that the association between specific insulin level and severity of coronary atherosclerosis lost its significance. CONCLUSIONS: The level of plasma fasting specific insulin was associated significantly with the severity of coronary atherosclerosis, as measured by Gensini score, but hyperinsulinemia showed no association with the severity of coronary atherosclerosis in multivariate analyses. PMID- 16477775 TI - Peripheral edema due to heart disease: diagnosis and outcome. AB - BACKGROUND: The purpose of this study was to determine whether baseline physical examination and history are useful in identifying patients with cardiac edema as defined by echocardiography, and to compare survival for patients with cardiac and noncardiac causes of edema. HYPOTHESIS: Physical examination and history data can help to identify patients with edema who have significant cardiac disease. METHODS: We reviewed the medical records of 278 consecutive patients undergoing echocardiography for evaluation of peripheral edema. We classified cardiac edema as the presence of any of the following: left ventricular ejection fraction < 45%, systolic pulmonary artery pressure > 45 mmHg, reduced right ventricular function, enlarged right ventricle, and a dilated inferior vena cava. RESULTS: The mean age of the 243 included patients was 67 +/- 12 years and 92% were male. A cardiac cause of edema was found in 56 (23%). Independent predictors of a cardiac cause of edema included chronic obstructive pulmonary disease (COPD, odds ratio [OR] 1.74, 95% confidence interval [CI] 1.14-2.60) and crackles (OR 1.98, 95% CI 1.26-3.10). The specificity for a cardiac cause of edema was high (91% for COPD, 93% for crackles); however, the sensitivity was quite low (27% for COPD, for 24% crackles). Compared with patients without a cardiac cause of edema, those with a cardiac cause had increased mortality (25 vs. 8% at 2 years, p < 0.01), even after adjustment for other characteristics (hazard ratio 1.55, 95% CI 1.08 2.24). CONCLUSIONS: A cardiac cause of edema is difficult to predict based on history and examination and is associated with high mortality. PMID- 16477776 TI - Effect of statins and white blood cell count on mortality in patients with ischemic left ventricular dysfunction undergoing percutaneous coronary intervention. AB - BACKGROUND: While morbidity and mortality were shown to be increased in the setting of an elevated white blood cell (WBC) count for patients with acute coronary syndrome, the impact of statin therapy on mortality for patients with an elevated WBC count is unknown in high-risk patients with coronary artery disease. HYPOTHESIS: The goal of this study was to determine whether statin therapy improved survival in patients with elevated WBC count undergoing percutaneous coronary intervention (PCI) with preexisting left ventricular (LV) dysfunction, a population at high risk for adverse outcomes. METHODS: We retrospectively evaluated consecutive patient procedures performed at our institution from 1996 through 1999. Patients had a technically adequate angiographic left ventriculogram with a calculated ejection fraction (EF) < or = 50%. Patients with prior coronary artery bypass graft were excluded. Mortality data were retrieved using the U.S. Social Security Death Index. Follow-up ranged from 3.5 to 6.5 years. Means are provided with +/- standard deviation, and p values < 0.05 were considered significant. RESULTS: Of the study population of 238 patients (average EF 39 +/- 9.8%, mean age 57.5 +/- 12 years, 68% men) 61% underwent PCI for a recent myocardial infarction, 68% received stents, and 65% were discharged on statins. Mean WBC count was 9,000 +/- 3,100 cells/mm3, with 28% of patients having a WBC > or = 10,000 cells/mm3. During follow-up, 27% of our population died. Patients with a WBC > or = 10,000 had worse survival than patients with WBC < 10,000 (1-year survival: 86 vs. 96%, p < 0.05; 3-year survival: 79 vs. 89%, p < 0.05). Survival was significantly improved in patients on statin therapy regardless of WBC count, but the greatest benefit tended to be in patients with WBC > or = 10,000 (WBC > or = 10,000; odds ratio [OR] 5.14, 95% confidence interval [CI] 1.44-19.0, WBC < 10,000; OR 2.79,95% CI 1.13-7.1). Proportional hazard regression analysis demonstrated that both statin therapy and WBC count predicted mortality. CONCLUSION: Patients undergoing PCI with LV dysfunction discharged on statins had improved survival regardless of WBC count, with a trend for greater improvement in patients with elevated WBC counts. In addition, WBC count predicts mortality in this high-risk population with LV dysfunction undergoing PCI. PMID- 16477777 TI - Images in cardiology: congenitally corrected transposition of the great arteries with dextrocardia. PMID- 16477778 TI - Ivan Petrovitch Pavlov. PMID- 16477779 TI - Oncology nurses of today mentor the cancer caregivers of tomorrow. PMID- 16477780 TI - Share what you know, be a mentor. PMID- 16477781 TI - Georgia nurses welcome oncology nurse from Uruguay. PMID- 16477782 TI - Confused about Medicare Part D? Check out these web sites. PMID- 16477783 TI - 2005 closes with little federal funding to support nurses or people with cancer. PMID- 16477784 TI - Meet ONS director-at-large Ellyn E. Matthews. PMID- 16477785 TI - Budget process sometimes involves making tough decisions. PMID- 16477786 TI - The future of cancer nursing. PMID- 16477787 TI - In our blood. PMID- 16477788 TI - Weighting for Godot. PMID- 16477789 TI - The Popeye principle: selling child health in the first nutrition crisis. AB - The cartoon character Popeye the Sailor was capable of superhuman feats of strength after eating a can of spinach. Popeye ate spinach because the association of spinach with strength was a product of the first national nutrition crisis in the United States: the 1920s fight against child malnutrition. Spanning the first three decades of the twentieth century, the malnutrition crisis arose from the confluence of many different events including the invention of nutrition science and new standards for height and weight; international food crises created by world war; the rise of consumerism, advertising, and new forms of mass media; and Progressive reformers' conviction that education was a key component of any solution. The history of the malnutrition crisis presented in this essay synthesizes disparate histories concerning advertising, public health, education, consumerism, philanthropy, and Progressive Era reform with original analysis of a major nutrition education program sponsored by the Commonwealth Fund in the 1920s. Because the character of Popeye came to embody one of the nutritional norms advocated in the 1920s, I refer to the influence of culturally constructed social norms on children's beliefs about health and nutrition as the Popeye Principle. The history of the malnutrition crisis demonstrates the importance of understanding the cultural and economic conditions surrounding childhood nutrition, the use and influence of numerical norms, and the mutually reinforcing influences on children's nutritional norms from their parents, peers, teachers, and culture. PMID- 16477790 TI - Obesity, courts, and the new politics of public health. AB - Health care politics are changing. They increasingly focus not on avowedly public projects (such as building the health care infrastructure) but on regulating private behavior. Examples include tobacco, obesity, abortion, drug abuse, the right to die, and even a patient's relationship with his or her managed care organization. Regulating private behavior introduces a distinctive policy process; it alters the way we introduce (or frame) political issues and shifts many important decisions from the legislatures to the courts. In this article, we illustrate the politics of private regulation by following a dramatic case, obesity, through the political process. We describe how obesity evolved from a private matter to a political issue. We then assess how different political institutions have responded and conclude that courts will continue to take the leading role. PMID- 16477791 TI - Weighing both sides: morality, mortality, and framing contests over obesity. AB - Despite recent and growing media attention surrounding obesity in the United States, the so-called obesity epidemic remains a highly contested scientific and social fact. This article examines the contemporary obesity debate through systematic examination of the claims and claimants involved in the controversy. We argue that four primary groups-antiobesity researchers, antiobesity activists, fat acceptance researchers, and fat acceptance activists-are at the forefront of this controversy and that these groups are fundamentally engaged in framing contests over the nature and consequences of excess body weight. While members of the fat acceptance groups embrace a body diversity frame, presenting fatness as a natural and largely inevitable form of diversity, members of the antiobesity camp frame higher weights as risky behavior akin to smoking, implying that body weight is under personal control and that people have a moral and medical responsibility to manage their weight. Both groups sometimes frame obesity as an illness, which limits blame by suggesting that weight is biologically or genetically determined but simultaneously stigmatizes fat bodies as diseased. While the antiobesity camp frames obesity as an epidemic to increase public attention, fat acceptance activists argue that concern over obesity is distracting attention from a host of more important health issues for fat Americans. We examine the strategies claimants use to establish their own credibility or discredit their opponents, and explain how the fat acceptance movement has exploited structural opportunities and cultural resources created by AIDS activism and feminism to wield some influence over U.S. public health approaches. We conclude that notions of morality play a central role in the controversy over obesity, as in many medical disputes, and illustrate how medical arguments about body weight can be used to stymie rights claims and justify morality-based fears. PMID- 16477792 TI - Public opinion and the politics of obesity in America. AB - Health policy experts have recently sounded the warning about the severe health and economic consequences of America's growing rates of obesity. Despite this fact, obesity has only begun to enter America's political consciousness and we have little information about what average Americans think of obesity or whether they support obesity-related policies. Using unique survey data collected by the authors, this essay examines public attitudes toward obesity and obesity policy. We find that, contrary to the views of health experts, most Americans are not seriously concerned with obesity, express relatively low support for obesity targeted policies, and still view obesity as resulting from individual failure rather than environmental or genetic sources. Given the absence of elite discourse on this problem, we also find that typical determinants of policy preferences, such as ideology or partisanship, are not good predictors of attitudes on obesity policy. Rather, with a low-valence issue such as obesity, the public utilizes other attitudinal frameworks such as their opinions on smoking policy and the environmental culpability for obesity. The implications of these findings for obesity policy and research on health-related public opinion are discussed. PMID- 16477793 TI - The chronicling of obesity: growing awareness of its social, economic, and political contexts. PMID- 16477795 TI - Qui tam actions: best practices for relator's counsel. AB - The False Claims Act empowers the U.S. Government to identify and prosecute fraud. It does so in no small part by engaging qui tam relators who, with their attorneys, are deputized under the Act to help investigate and prosecute these cases. By combining the insight and industry-specific knowledge of the citizenry with the investigative and prosecutorial resources of the government, Congress attempted to facilitate a united front against pervasive fraud and abuse by government contractors, an aggressive effort that has borne fruit, recovering funds and protecting important federal programs. This Article proposes ways in which relators' counsel and government attorneys can work together more effectively in these cases to achieve the goals of the statute. PMID- 16477796 TI - Medicare and America's healthcare system in transition: from the death of managed care to the Medicare Modernization Act of 2003 and beyond. AB - This Article traces the transition--in Medicare, specifically, and in the American healthcare system, generally--from the aftermath of the Balanced Budget Act of 1997 to the passage of the Medicare Modernization Act of 2003. During this time, restrictive managed care died under an onslaught of resurgent cost pressures, legislative and legal attacks, and a vehement physician and consumer backlash. The subsequent reversion to more generous (and more expensive) health plans coincided with a recession in 2001 to trigger a return to rapidly escalating healthcare spending and yet another in the Nation's series of healthcare crises. Current trends suggest that future policymakers will have no choice but to confront the consequences of rapidly rising rates of healthcare spending. PMID- 16477797 TI - Death of an exception or much ado about nothing. AB - Hospitals often engage in physician recruitment in an effort to fulfill a community need for a particular medical specialty. In doing so, the hospital must comply with the regulatory requirements of the physician recruitment exception of the Stark law, which over the years has generated a great deal of discussion and perhaps confusion. The publication of the Stark II, Phase II regulations in March 2004 was supposed to provide guidance and clarity, but the new regulations have raised a number of new issues and concerns, particularly regarding the requirements imposed on recruiting arrangements involving group practices. This Article reviews the regulatory requirements of the new physician recruitment exception and addresses several of the concerns that have been raised. Specifically, it examines the new regulatory definition of the "geographic area served by the hospital," the restrictions on income guarantees when the recruited physician joins a group practice, and the prohibition on additional practice restrictions. The author concludes that, while some of these concerns are legitimate, others will have little practical implication and should not hinder the ability of hospitals to engage in reasonable, beneficial recruitment activities. PMID- 16477798 TI - Fiduciary duties of interlocking directors within a nonprofit health system. AB - In response to regulatory and financial pressures, entities participating in the healthcare industry have joined with others in order to maintain, even improve, their market positions. In the non-profit sector of the industry, partnerships, mergers, and acquisitions have included arrangements whereby some corporate partners have interlocking directors. After review of the fiduciary duties of care and loyalty owed by corporate board members, and their application to traditional performance and conflict of interest situations, the authors address two scenarios raising interlocking director issues. PMID- 16477799 TI - Balancing in a crisis? Bioterrorism, public health and privacy. AB - Post-September 11, the government has been rapidly funding public health initiatives to bolster the Nation's ability to respond to bioterrorist attacks. While the infusion of money into the public health system is laudable, the pressure to enact legislation quickly has resulted in laws and policies that ignore privacy and civil liberties and that favor anti-bioterror initiatives over more common public health concerns. A public health agenda that ignores privacy and civil liberties will undermine public trust, leading people to not fully participate in critical public health activities. Our Nation is far more likely to succeed in preventing and responding to a potential act of bioterrorism if we embrace the principle that advancing public health and preserving individual liberties are symbiotic and inextricable. PMID- 16477800 TI - EMTALA Compliance Checklist. PMID- 16477801 TI - The fate and intermediary metabolism of stearic acid. AB - Coming from the Greek for "hard fat," stearic acid represents one of the most abundant FA in the Western diet. Otherwise known as n-octadecanoic acid (18:0), stearate is either obtained in the diet or synthesized by the elongation of palmitate, the principal product of the FA synthase system in animal cells. Stearic acid has been shown to be a very poor substrate for TG synthesis, even as compared with other saturated fats such as myristate and palmitate, and in human studies stearic acid has been shown to generate a lower lipemic response than medium-chain saturated FA. Although it has been proposed that this may be due to less efficient absorption of stearic acid in the gut, such findings have not been consistent. Along with palmitate, stearate is the major substrate for the enzyme stearoyl-CoA desaturase, which catalyzes the conversion of stearate to oleate, the preferred substrate for the synthesis of TG and other complex lipids. In mice, targeted disruption of the stearoyl-CoA desaturase-1 (SCD1) gene results in the generation of a lean mouse that is resistant to diet-induced obesity and insulin resistance. SCD1 also has been shown to be a key target of the anorexigenic hormone leptin, thus underscoring the importance of this enzyme, and consequently the cellular stearate-to-oleate ratio, in lipid metabolism and potentially in the treatment of obesity and related disorders. PMID- 16477803 TI - Effects of stearic acid on plasma lipid and lipoproteins in humans. AB - More than 40 years ago, saturated FA with 12, 14, and 16 carbon atoms (lauric acid, myristic acid, and palmitic acid) were demonstrated to be "hypercholesterolemic saturated FA". It was further concluded that the serum total cholesterol level would hardly be changed by isocaloric replacement of stearic acid (18:0) by oleic acid (cis-18:1n-9) or carbohydrates. These earlier studies did not address the effects of the various FA on the serum lipoprotein profile. Later studies found that the hypercholesterolemic saturated FA increase serum total cholesterol levels by raising concentrations of both the atherogenic LDL and the antiatherogenic HDL. Consequently, the ratio of total to HDL cholesterol will hardly change when carbohydrates replace these saturated FA. Compared with other saturated FA, stearic acid lowers LDL cholesterol. Studies on the effects on HDL cholesterol are less conclusive. In some, the effects on HDL cholesterol were comparable to those of palmitic acid, oleic acid, and linoleic acid, whereas in others a decrease was observed. This may suggest that in this respect the source of stearic acid is of importance, which needs however further study. From all these studies, however, it can be concluded that stearic acid may decrease the ratio of total to HDL cholesterol slightly when compared with palmitic or myristic acid. Without doubt, the effects of stearic acid are more favorable than those of trans monounsaturated FA. PMID- 16477802 TI - Dietary stearic acid and risk of cardiovascular disease: intake, sources, digestion, and absorption. AB - Individual FA have diverse biological effects, some of which affect the risk of cardiovascular disease (CVD). In the context of food-based dietary guidance designed to reduce CVD risk, fat and FA recommendations focus on reducing saturated FA (SFA) and trans FA (TFA), and ensuring an adequate intake of unsaturated FA. Because stearic acid shares many physical properties with the other long-chain SFA but has different physiological effects, it is being evaluated as a substitute for TFA in food manufacturing. For stearic acid to become the primary replacement for TFA, it is essential that its physical properties and biological effects be well understood. PMID- 16477804 TI - Influence of dietary saturated fatty acids on the regulation of plasma cholesterol concentration. AB - The specific effects of individual fatty acids (FA) on plasma cholesterol levels, in the range habitually consumed by humans, on plasma cholesterol levels is not usually presented by the literature. Conclusions have been made regarding the cholesterolemic effect of individual FA, even though these FA cannot be tested individually. It appears that FA balance of the diet may be more important than individual FA intakes. Variation in plasma cholesterol response to diet is influenced by many factors, such as gene-nutrient interactions. The effect on human health of current processes used in the food industry that are certain to change dietary fat composition and TG structure is yet to be fully explored. Some of the relevant research regarding dietary fat and plasma cholesterol levels is reviewed. PMID- 16477806 TI - Influence of stearic acid on postprandial lipemia and hemostatic function. AB - It has been suggested that fats rich in stearic acid may result in exaggerated postprandial lipemia and have adverse effects on hemostatic function. The effects of test meals containing different saturated and monounsaturated FA were compared in healthy subjects in a series of studies to investigate this hypothesis. Stearic acid, when present as cocoa butter, resulted in similar postprandial lipemia and factor VII activation compared with a meal containing high-oleic sunflower oil. Stearic acid when presented as shea butter or as randomized stearate-rich TAG resulted in decreased postprandial lipemia and decreased postprandial activation of factor VII. Stearic acid-rich test meals did not result in impaired fibrinolytic activity compared with either a low-fat meal or a meal high in oleate. The difference in responses between the different stearic acid-rich fats appears to be due to varying solid fat contents of the fats at 37 degrees C. PMID- 16477805 TI - Overview of hemostatic factors involved in atherosclerotic cardiovascular disease. AB - Hemostatic factors associated with the development of cardiovascular disease (CVD) include fibrinogen, von Willebrand factor, tissue plasminogen activator (tPA) antigen, plasminogen activator inhibitor-1 (PAI-1), and factor VII. Each SD increment of these increases the association by 24-30%. Most hemostatic factors are intercorrelated with inflammatory markers [e.g., C-reactive protein (CRP)] and LDL cholesterol. Fibrinogen seems the most fundamental hemostatic risk factor for CVD. The Framingham Study reaffirms the significant linear risk factor trends across fibrinogen tertiles (P< 0.001) for age, body mass index, smoking, diabetes mellitus, total cholesterol, HDL cholesterol, and TG in both sexes. Fibrinogen may also directly increase CVD risk because of its role in platelet aggregation, plasma viscosity, and fibrin formation. Fibrinogen is also an acute-phase reactant that is elevated in inflammatory states. Fibrinogen mediates the thrombogenic effect of other risk factors. Fibrinogen levels increase with the number of cigarettes smoked and quickly fall after smoking cessation. This rapid fibrinogen decline may be a mechanism for CVD risk reduction after smoking cessation. Weight loss is accompanied by reduced fibrinogen. The correlation between fibrinogen and LDL cholesterol suggests that lipid-imposed CVD risk is mediated partly through fibrinogen. Hyperreactive platelets of diabetics may result in part from their increased fibrinogen. Elevated fibrinogen and CRP of unstable angina suggest an acute-phase reaction. Prevalence, case-control, angiographic, and echocardiogram investigations incriminate hemostatic and inflammatory markers as strong independent risk factors for initial and recurrent CVD. Framingham Study data indicate that each SD increase in fibrinogen imposes a 20% independent increment in risk. It may be concluded that fibrinogen and CRP determination may be useful screening tools to identify individuals at added risk for thrombotic complications of CVD. PMID- 16477807 TI - Influence of stearic acid on hemostatic risk factors in humans. AB - Stearic acid has been claimed to be prothrombotic. Elevated plasma factor VII coagulant activity (FVIIc) may raise the risk of coronary thrombosis in the event of plaque rupture. Fibrinogen, an acute-phase protein, is necessary for normal blood clotting; however, elevated levels of fibrinogen increase the risk of coronary heart disease (CHD). Here I report the results of three controlled, human dietary intervention studies, which used a randomized crossover design to investigate the hemostatic effects of stearic acid-rich test diets in healthy young men. A diet high in stearic acid (shea butter) resulted in a 13% lower fasting plasma FVIIc than a high palmitic acid diet, and was 18% lower than a diet high in myristic and lauric acids (P = 0.001) after 3 wk of intervention. The stearic acid-rich test fat increased plasma fibrinogen concentrations slightly compared with the myristic-lauric acid diet (P < 0.01). When investigating the acute effects of fatty meals, those high in stearic acid (synthesized test fat) resulted in a smaller postprandial increase in FVII than those high in trans and oleic FA, indicating a smaller increase in activated FVII after ingesting stearic acid compared with fats high in monounsaturated FA, probably caused by lower postprandial lipemia. Thus, the present investigations did not find dietary stearic acid to be more thrombogenic, in either fasting effects compared with other long-chain FA, or in acute effects compared with dietary unsaturated FA, including trans monounsaturated FA. The slightly increased effect on fasting plasma fibrinogen may be biologically insignificant, but it should be investigated further. PMID- 16477808 TI - Mitochondrial cholesterol transport: a possible target in the management of hyperlipidemia. AB - Sterol 27-hydroxylase (CYP27A1) may defend cells against accumulation of excess cholesterol, making this enzyme a possible target in the management of hyperlipidemia. The study objective was to analyze cholesterol homeostatic responses to increases in CYP27A1 activity in HepG2 cells and primary human hepatocytes. Increasing CYP27A1 activity by increasing enzyme expression led to significant increases in bile acid synthesis with compensatory increases in HMG CoA reductase (HMGR) activity/protein, LDL receptor (LDLR) mRNA, and LDLR mediated cholesterol uptake. Under these conditions, only a small increase in cellular 27-hydroxycholesterol (27OH-Chol) concentration was observed. No changes were detected in mature sterol regulatory element-binding proteins (SREBP) 1 or 2. Increasing CYP27A1 activity by increasing mitochondrial cholesterol transport (i.e., substrate availability) led to greater increases in bile acid synthesis with significant increases in cellular 27OH-Chol concentration. Mature SREBP 2 protein decreased significantly with compensatory decreases in HMGR protein. No change was detected in mature SREBP 1 protein. Despite increasing 27OH-Chol and lowering SREBP 2 protein concentrations, LDLR mRNA increased significantly, suggesting alternative mechanisms of LDLR transcriptional regulation. These findings suggest that regulation of liver mitochondrial cholesterol transport represents a potential therapeutic strategy in the treatment of hyperlipidemia and atherosclerosis. PMID- 16477810 TI - Analogs of squalene and oxidosqualene inhibit oxidosqualene cyclase of Trypanosoma cruzi expressed in Saccharomyces cerevisiae. AB - Recently, a number of inhibitors of the enzyme oxidosqualene cyclase (OSC; EC 5.4.99.7), a key enzyme in sterol biosynthesis, were shown to inhibit in mammalian cells the multiplication of Trypanosoma cruzi, the parasite agent of Chagas' disease. The gene coding for the OSC of T. cruzi has been cloned and expressed in Saccharomyces cerevisiae. The expression in yeast cells could be a safe and easy model for studying the activity and the selectivity of the potential inhibitors of T. cruzi OSC. Using a homogenate of S. cerevisiae cells expressing T. cruzi OSC, we have tested 19 inhibitors: aza, methylidene, vinyl sulfide, and conjugated vinyl sulfide derivatives of oxidosqualene and squalene, selected as representative of different classes of substrate analog inhibitors of OSC. The IC50 values of inhibition (the compound concentration at which the enzyme is inhibited by 50%) are compared with the values obtained using OSC of pig liver and S. cerevisiae. Many inhibitors of pig liver and S. cerevisiae OSC show comparable IC50 for T. cruzi OSC, but some phenylthiovinyl derivatives are 10-100 times more effective on the T. cruzi enzyme than on the pig or S. cerevisiae enzymes. The expression of proteins of pathogenic organisms in yeast seems very promising for preliminary screening of compounds that have potential therapeutic activity. PMID- 16477811 TI - Fatty acid and hydroxy acid adaptation in three gram-negative hydrocarbon degrading bacteria in relation to carbon source. AB - The lipids of three gram-negative bacteria, Acinetobacter calcoaceticus, Marinobacter aquaeolei, and Pseudomonas oleovorans grown on mineral media supplemented with ammonium acetate or hydrocarbons, were isolated, purified, and their structures determined. Three pools of lipids were isolated according to a sequential procedure: unbound lipids extracted with organic solvents, comprising metabolic lipids and the main part of membrane lipids, OH--labile lipids (mainly ester-bound in the lipopolysaccharides, LPS) and H+-labile lipids (mainly amide bound in the LPS). Unsaturated FA composition gave evidence for an aerobic desaturation pathway for the synthesis of these acids in A. calcoaceticus and M. aquaeolei, a nonclassic route in gram-negative bacteria. Surprisingly, both aerobic and anaerobic pathways are operating in the studied strain of P. oleovorans. The increase of the proportion of saturated FA observed for the strain of P. oleovorans grown on light hydrocarbons would increase the temperature transition of the lipids for maintaining the inner membrane fluidity. An opposite phenomenon occurs in A. calcoaceticus and M. aquaeolei grown on solid or highly viscous C19 hydrocarbons. The increases of FA < C18 when the bacteria were grown on n-nonadecane, or of iso-FA in cultures on isononadecane would decrease the transition temperature of the lipids, to maintain the fluidity of the inner membranes. Moreover, P. oleovorans grown on hydrocarbons greatly decreases the proportion of P-hydroxy acids of LPS, thus likely maintaining the physical properties of the outer membrane. By contrast, no dramatic change in hydroxy acid composition occurred in the other two bacteria. PMID- 16477809 TI - Effect of dietary n-6 and n-3 polyunsaturated fatty acids on peroxidizability of lipoproteins in steers. AB - The susceptibility of major plasma lipoproteins to lipoperoxidation was studied in relation to the FA composition of their neutral and polar lipids in steers given PUFA-rich diets. Two trials used, respectively, 18 ("sunflower" experiment, S) or 24 ("linseed" experiment, L) crossbred Salers x Charolais steers. Each involved three dietary treatments over a 70-d period: a control diet (CS or CL diets) consisting of hay and concentrate, or the same diet supplemented with oilseeds (4% diet dry matter) fed either as seeds (SS or LS diets) or continuously infused into the duodenum (ISO or ILO diets). Compared with control diets, ISO and ILO treatments tended to decrease the resistance time of LDL and HDL classes to peroxidation, mainly owing to the enrichment of their polar and neutral lipids with PUFA. With diets SS and LS, sensitivity of major lipoprotein classes (LDL, light and heavy HDL) was not affected because ruminal hydrogenation of dietary PUFA decreased their incorporation into lipoparticles. ISO and ILO treatments induced a more important production of conjugated dienes and hydroperoxides generated by peroxidation in the three lipoprotein classes due to the higher amounts of PUFA esterified in lipids of the core and the hydrophilic envelope of particles. The production of malondialdehyde (MDA) increased in steers fed linseed supplements, indicating that MDA production did not occur with linoleic acid provided by sunflower oil supplements. Thus, plasma peroxidation of PUFA generates toxic products in steers fed diets supplemented with PUFA and can be deleterious for the health of the animal during long-term treatment. PMID- 16477813 TI - Esophageal Doppler monitoring during organ donor resuscitation: new benefits of existing technology. PMID- 16477812 TI - Lipolysis of different oils using crude enzyme isolate from the intestinal tract of rainbow trout, Oncorhynchus mykiss. AB - Crude enzyme isolate was prepared from the intestine of rainbow trout. Positional specificity of the crude enzyme isolate was determined from both 1(3)- and 2-MAG products after in vitro lipolysis of radioactive-labeled triolein. The ratio of 2 MAG/1(3)-MAG was 2:1, suggesting that the overall lipase specificity of the enzyme isolate from rainbow trout tended to be 1,3-specific; however, activity against the sn-2 position also was shown. In vitro lipolysis of four different unlabeled oils was performed with the crude enzyme isolate. The oils were: structured lipid [SL; containing the medium-chain FA (MCFA) 8:0 in the sn-1,3 positions and long-chain FA (LCFA) in the sn-2 position], DAG oil (mainly 1,3 DAG), fish oil (FO), and triolein (TO). MCFA were rapidly hydrolyzed from the SL oil. LCFA including n-3 PUFA were, however, preserved in the sn-2 position and therefore found in higher amounts in 2-MAG of SL compared with 2-MAG of FO, DAG, and TO. Lipolysis of the DAG oil produced higher amounts of MAG than the TAG oils, and 1(3)-MAG mainly was observed after lipolysis of the DAG oil. The positional specificity determined and the results from the hydrolysis of the different oils suggest that n-3 very long-chain PUFA from structured oils may be used better by aquacultured fish than that from fish oils. PMID- 16477814 TI - Pedophiles and stalkers as transplant candidates: one program's experience. AB - Transplant assessment of pedophiles and stalkers presents a challenge for clinicians and requires the ability to separate the science from the hysteria. Although rarely seen in organ transplant clinics, the presence of psychosexual disorders can result in both criminal charges and harsh societal reactions. Clinicians must be able to assess and make informed recommendations about the suitability of the candidate and any potential risks for staff, other patients, and society at large. Six patients (5 pedophiles and 1 stalker) were assessed at our major regional multiorgan transplant program; each patient presented with unique and challenging assessment, treatment, and selection issues for the transplant team. The objectives of this article are to raise awareness of the issues associated with pedophiles and stalkers, and to provide the basis for better-informed discussions based on one program's struggles with its candidate selection and follow-up. PMID- 16477815 TI - Dollars and sense of successful rehabilitation. AB - Improving employment outcomes for transplant recipients can positively contribute to a patient's identity, self-esteem, and quality of life. Placing a dollar figure on the costs associated with advanced medical procedures is much easier than placing a dollar figure on the benefits resulting from being employed after transplantation. This article outlines the work of the Life Options Rehabilitation Advisory Council and how it has assisted people with kidney disease-those on dialysis and those who received transplants-to achieve life goals and provide long-term access to employer group health plans. A review of current work incentives includes the Ticket to Work and the Work Incentives Improvement Act, Medicaid Buy-in Programs, and other work incentives related to the Social Security system. Effective rehabilitation outcomes can be enhanced in the transplant population through knowledge of work incentives and awareness of obstacles that are perceived by rehabilitation agencies in working with this group. PMID- 16477816 TI - Differences in quality of life before transplantation among transplant recipients with respect to selected socioeconomic variables. AB - PURPOSE: To examine differences in quality of life before transplantation among transplant recipients with respect to a selected set of socioeconomic variables related to household/housing, family social and health history, and availability and use of community resources. METHODS: An exploratory-descriptive study in a US university's transplant clinic. Sample included 249 kidney, liver, and pancreas transplant recipients aged 18 years or older. Instruments were a socioeconomic data questionnaire, the Sickness Impact Profile, the Adult Self-Image Scale, the Quality of Life Index, and a general quality of life scale. Descriptive statistics and analysis of variance with Bonferroni adjustment were used with a significance level of .05. Effect sizes were estimated. RESULTS: Quality of life did not differ significantly for any variable except for the use of private insurance combined with public health insurance. Such coverage was associated with significantly better scores on the psychosocial and total dimensions of the Sickness Impact Profile, indicating better health before transplantation. CONCLUSION: Researchers must first explore why a mix of private and public health insurance is associated with a higher quality of life and second reexamine whether these results occur at other transplant centers. These results also provide direction for social work professionals as they counsel patients in an attempt to help patients achieve a better psychosocial and physical quality of life before transplantation. PMID- 16477817 TI - Liver transplant recipients' first year of posttransplant recovery: a longitudinal study. AB - A longitudinal study of 20 liver transplant recipients was conducted to investigate their posttransplant recovery experience. Data were collected using semistructured interviews at 6 weeks, 6 months, and 1 year after transplantation. Qualitative analysis of data revealed physical, psychological, social, economic, and spiritual dimensions of recovery. Findings reflect ongoing improvement of physical health and functionality for most recipients. Those with continuing health problems often suffered from preexisting health conditions. Psychological adjustment was uneven, with intermittent periods of fear, anxiety, and depression. Some recipients reported short-lived split identities and personality changes. Social support of family was critical in the hospital and at home. Economic issues became primary by the 1-year interview, with all recipients questioning whether they could afford ongoing healthcare and medicines. Spiritual needs were met in secular and nonsecular activities. Findings suggest that healthcare personnel should attend to the lived experience of liver transplant recipients. PMID- 16477818 TI - Social work interventions with living related liver donors: the implications for practice. AB - Living related liver donations are increasingly used to address the lack of organs available to patients faced with end-stage liver disease. A significant need exists for enhanced psychosocial intervention for donors. We sought to explore donor reactions by assessing psychosocial issues after donation. A descriptive review of donors was undertaken. The overall incidence of psychosocial issues after donation was examined by using a uniform interview questionnaire and by anecdotal case studies. Responses tend to indicate a general positive reaction to donation; yet, findings suggest a need for more preparation in the predonor stage, 100% social work follow-up after donation to provide needed support services, and inclusion of the donor as a critical partner in organ transplant treatment. A more extensive study drawing on the questionnaire may offer a standardized approach to the treatment of organ donors. PMID- 16477819 TI - Transfer versus transition: success in pediatric transplantation brings the welcome challenge of transition. AB - CONTEXT: Increasing success with solid organ transplantation in children has increased the numbers of adolescents and young adults who are at an age to transfer to adult healthcare. OBJECTIVE: To determine the nature of transfer/transition of adolescents and young adults to adult healthcare. DESIGN: Using a qualitative approach, 24 young adults provided answers to 12 questions about their transfer to adult healthcare. Responses were compared to identify themes. SETTING: Subjects had been pediatric patients at Children's Hospital of Pittsburgh. Most were transferred to the University of Pittsburgh Medical Center. PATIENTS: The subjects had received, or were on the list to receive, solid organ transplants and had been transferred within 18 months of the study. RESULTS: Twenty-four of 37 potential subjects completed interviews. Those who participated were enthusiastic about making a contribution to the transfer process. Most subjects cited difficulty leaving the pediatric system. Challenges included abrupt transfers, insufficient communication between pediatric and adult specialty providers, and becoming "lost to follow-up." However, there was a consistent, clear statement that subjects preferred being treated as adults. They liked healthcare staff addressing them instead of their parents, being spoken to in an adult manner, and being responsible for their own care. Suggestions for improvement included early introduction of the concept of transition to adult care, providing written material about the transition process, and use of pediatric/adult social work collaboration to provide support for young adults in transition. PMID- 16477820 TI - Informed consent. AB - In the last half century, informed consent has become a central tenet of all research involving human subjects. However, even after the worldwide adoption of regulations aimed at ensuring the protection of subjects, some abuses continue. The installation of oversight bodies, such as institutional review boards, has prevented the bulk of deliberate mistreatment; however, unintentional coercion is still a problem. Informed consent is not limited to obtaining a signature on a consent form, but a process of decision making designed to protect the rights of patients and subjects. It is a constant, ongoing exchange of information between researcher and subject. Although research is invaluable to the advancement of medicine and transplantation, researchers have the responsibility to protect their subjects' rights and autonomy. This paper provides an overview of informed consent, principles behind the process, and implications of current regulations to assist readers in their endeavor for sound and ethical research. PMID- 16477821 TI - Research to practice: a national consensus conference. AB - PURPOSE: To gain consensus on aspects of the process of gaining consent for organ donation that should be mainstream daily practice. METHODS: A 3-day consensus conference of transplant professionals that provided a forum for research and innovative ideas about gaining consent for organ donation. Four work groups were assembled to address issues of gaining consent from organ and tissue donors: (1) demystifying first-person consent (donor authorization): is it a matter of law?, (2) recovery coordinators: getting from green to great, (3) maximizing the process: old views and new, and (4) donor family support: mother or smother? RESULTS: Participants reached a consensus about major consent-related practices, identified areas of practice variance, and defined topics in need of further research. CONCLUSIONS: The conference participants agreed that (1) the primary role of the organ procurement organization is to recover organs for transplantation from deceased donors and to facilitate distribution of those organs to the appropriate recipients; (2) early referral of potential donors from the hospital to the organ procurement organization is desirable; (3) a score less than 5 on the Glasgow Coma Scale should initiate end-of-life discussions with family members; a procurement coordinator from the organ procurement organization who is physically housed within the hospital is the most successful model for accommodating early referral of potential donors and optimizing family support practices; and (3) a decedent's right to donate should take precedence in the donation process and such designation by the donor before death is irrevocable and does not require the consent or concurrence of any person after the donor's death. PMID- 16477822 TI - Kidney graft survival in Italy and factors influencing it. AB - PURPOSE: National registry data are often a suitable basis for examination of transplant outcomes. Using data supplied by the Italian National Transplant Registry, established in 1995, we performed the first nationwide analysis of this kind. METHODS: A retrospective analysis of 4893 recipients of cadaveric kidneys transplanted in all Italian centers from 1995 through 2000 was done to study 5 year graft survival. The association between some donor and recipient variables and outcomes in renal transplantation was analyzed. Graft survival was 93% at 3 months, 89% at 1 year, 82% at 3 years, and 80% at 5 years after transplantation. RESULTS: A significant association between graft survival and donor age (old vs young, relative risk [RR] = 1.62, 95% CI 1.27-2.06) and recipient age (old vs young, RR = 1.25, 95% CI 1.02-1.53). Graft survival was also associated with cold ischemia time (24-36 hours, RR= 1.39, 95% CI 1.05-1.85 and >36 hours, RR= 1.94, 95% CI 1.32-2.86 vs 0-24 hours) and donor/recipient sex mismatch (female/male vs male/male, RR= 1.50, 95% CI 1.17-1.93). CONCLUSION: The quality of kidney transplantation in Italy is satisfactory and is comparable to that in other developed countries. Furthermore, our experience confirms that both donor and recipient factors are major determinants of renal allograft function. PMID- 16477823 TI - An exploratory investigation of quality of life in adult liver transplant recipients. AB - Quality of life is an important outcome indicator of experiences associated with liver transplantation. Unfortunately, quality of life has been defined in many different ways, causing confusion and misconceptions among practitioners, researchers, policy makers, and patients. This exploratory qualitative study was initiated to gain greater understanding regarding the adult transplant recipients' experiences with liver transplantation in order to direct future studies with this population and to assist in selection of a relevant quality of life survey tool for quantitative investigation. Twelve informants (7 women, 5 men) shared their experiences in face-to-face interviews conducted by the researcher. Analysis of these interviews reveals the importance of physiologic, psychological, social, spiritual, family, and socioeconomic aspects of quality of life for liver transplant recipients. PMID- 16477824 TI - West Nile virus: a challenge for transplant programs. AB - West Nile virus is a new challenge for transplant programs worldwide. It is a mosquito-borne disease, which has become increasingly prevalent in North America since it was first recognized in New York in 1999. A review of a case study and the literature shows that the morbidity and mortality associated with West Nile virus infection and transplant recipients are alarmingly high. Treatment options are limited because of transplant programs' limited experience in working with this virus. Transplant programs must develop action plans for education on West Nile virus and its prevention to decrease the risk of infection among their transplant recipients. PMID- 16477825 TI - The Georgia Transplant Foundation. PMID- 16477826 TI - Optics of the average normal cornea from general and canonical representations of its surface topography. AB - Generally, the analysis of corneal topography involves fitting the raw data to a parametric geometric model that includes a regular basis surface, plus some sort of polynomial expansion to adjust the more irregular residual component. So far, these parametric models have been used in their canonical form, ignoring that the observation (keratometric) coordinate system is different from corneal axes of symmetry. Here we propose, instead, to use the canonical form when the topography is referenced to the intrinsic corneal system of coordinates, defined by its principal axes of symmetry. This idea is implemented using the general expression of an ellipsoid to fit the raw data given by the instrument. Then, the position and orientation of the three orthogonal semiaxes of the ellipsoid, which define the intrinsic Cartesian system of coordinates for normal corneas, can be identified by passing to the canonical form, by standard linear algebra. This model has been first validated experimentally obtaining significantly lower values for rms fitting error as compared with previous standard models: spherical, conical, and biconical. The fitting residual was then adjusted by a Zernike polynomial expansion. The topographies of 123 corneas were analyzed obtaining their radii of curvature, conic constants, Zernike coefficients, and the direction and position of the optical axis of the ellipsoid. The results were compared with those obtained using the standard models. The general ellipsoid model provides more negative values for the conic constants and lower apex radii (more prolate shapes) than the standard models applied to the same data. If the data are analyzed using standard models, the resulting mean shape of the cornea is consistent with previous studies, but when using the ellipsoid model we find new interesting features: The mean cornea is a more prolate ellipsoid (apical power 50 D), the direction of the optical axis is about 2.3 degrees nasal, and the residual term shows three Zernike coefficients significantly higher than zero (third-order trefoil and fourth- and sixth-order spherical). These three nonzero Zernike coefficients are responsible for most of the higher-order aberrations of the average cornea. Finally, we propose and implement a simple method for three dimensional registration of corneal topographies, passing from the general to the canonical form of the ellipsoid. PMID- 16477827 TI - Time course of brightness under transient glare condition. AB - It was shown that a peripheral glare source reduces the brightness of a foveal stimulus. We hypothesized that this brightness reduction is governed by an inhibitory effect of the glare source on the test. We reported the results of an investigation of the dynamic of brightness reduction of an incremental stimulus immediately after the onset of a glare source in the field of view. A magnitude comparison paradigm using constant stimuli was adopted to determine the luminance that appeared equal in brightness to the standard patch. The luminance of the standard stimulus was in the mesopic range (0.5 cd/m2), and the levels of glare were 15 and 60 lx. Results showed that the time course of brightness reduction followed the typical shape attributed to the Broca-Sulzer effect. Data were fitted with a model that first considers the response of a peripheral ganglion cell to glare and then its inhibitory effect on the test signals. We discussed the plausibility of a postretinal stage of processing. PMID- 16477828 TI - Binocular interactions in random chromatic changes at isoluminance. AB - To examine the type of chromatic interactions at isoluminance in the phenomenon of binocular vision, I have determined simple visual reaction times (VRT) under three observational conditions (monocular left, monocular right, and binocular) for different chromatic stimuli along random color axes at isoluminance (simultaneous L-, M-, and S-cone variations). Upper and lower boundaries of probability summation as well as the binocular capacity coefficient were estimated with observed distributions of reaction times. The results were not consistent with the notion of independent chromatic channels between eyes, suggesting the existence of excitatory and inhibitory binocular interactions at suprathreshold isoluminance conditions. PMID- 16477829 TI - Processing halftone color images by vector space methods. AB - The reproduction of color images by color halftoning can be characterized by the Neugebauer model/equation. However, the Neugebauer equation is not easy to solve because of the highly nonlinear relationship between the underlying Neugebauer primaries and the colorants. We attempt to solve the Neugebauer equation by vector space methods. The proposed method of solution is applicable to any number of colorants, although our experimental results are confined to the CMY and CMYK cases. Among the constraints we consider are those related to a bound on the permissible amount of total ink and a bound on the total cost of applying colorants to achieve a satisfactory level of color reproduction. Our results demonstrate that the vector space method is a feasible approach for solving for the required amounts of colorants in the constrained color halftoning problem. PMID- 16477830 TI - Image reconstruction: a unifying model for resolution enhancement and data extrapolation. Tutorial. AB - In reconstructing an object function F(r) from finitely many noisy linear functional values integral of F(r)Gn(r)dr we face the problem that finite data, noisy or not, are insufficient to specify F(r) uniquely. Estimates based on the finite data may succeed in recovering broad features of F(r), but may fail to resolve important detail. Linear and nonlinear, model-based data extrapolation procedures can be used to improve resolution, but at the cost of sensitivity to noise. To estimate linear-functional values of F(r) that have not been measured from those that have been, we need to employ prior information about the object F(r), such as support information or, more generally, estimates of the overall profile of F(r). One way to do this is through minimum-weighted-norm (MWN) estimation, with the prior information used to determine the weights. The MWN approach extends the Gerchberg-Papoulis band-limited extrapolation method and is closely related to matched-filter linear detection, the approximation of the Wiener filter, and to iterative Shannon-entropy-maximization algorithms. Non linear versions of the MWN method extend the noniterative, Burg, maximum-entropy spectral-estimation procedure. PMID- 16477832 TI - Methods to detect objects in photon-limited images. AB - We investigate the problem of detecting and localizing a known signal in a photon limited image, where Poisson noise is the dominant source of image degradation. For this purpose we developed and evaluated three new algorithms. The first two are based on the impulse restoration (IR) principle and the third is based on the generalized likelihood ratio test (GLRT). In the IR approach, the problem is formulated as one of restoring a delta function at the location of the desired object. In the GLRT approach, which is a well-known variation on the optimal likelihood ratio test, the problem is formulated as a hypothesis testing problem, in which the unknown background intensity of the image and the intensity scale of the object are obtained by maximum-likelihood estimation. We used Monte Carlo simulations and localization receiver operating characteristic (LROC) curves to evaluate the proposed algorithms quantitatively. LROC curves demonstrate the ability of an algorithm to detect and locate objects in a scene correctly. Our simulations demonstrate that the GLRT approach is superior to all other tested algorithms. PMID- 16477831 TI - Optical image processing using the photoinduced anisotropy of pyrrylfulgide. AB - A synthesized photochromic compound-pyrrylfulgide--is prepared as a thin film doped in a polymethylmethacrylate (PMMA) matrix. Under irradiation by UV light, the film converts from the bleached state into a colored state that has a maximum absorption at 635 nm and is thermally stable at room temperature. When the colored state is irradiated by a linearly polarized 650 nm laser, the film returns to the bleached state; photoinduced anisotropy is produced during this process. Application of optical image processing methods using the photoinduced anisotropy of the pyrrylfulgide/PMMA film is described. Examples in non-Fourier optical image processing, such as contrast reversal and image subtraction and summation, as well as in Fourier optical image processing, such as low-pass filtering and edge enhancement, are presented. PMID- 16477833 TI - Multiresolution phase retrieval in the fresnel region by use of wavelet transform. AB - A multiresolution (multiscale) analysis based on wavelet transform is applied to the problem of optical phase retrieval from the intensity measured in the in-line geometry (lens-free). The transport-of-intensity equation and the Fresnel diffraction integral are approximated in terms of a wavelet basis. A solution to the phase retrieval problem can be efficiently found in both cases using the multiresolution concept. Due to the hierarchical nature of wavelet spaces, wavelets are well suited to multiresolution methods that contain multigrid algorithms. Appropriate wavelet bases for the best solution approximation are discussed. The proposed approach reduces the computational complexity and accelerates the convergence of the solution. It is robust and reliable, and successful on both simulated and experimental images obtained with hard x rays. PMID- 16477834 TI - Wavefront reconstruction from its gradients. AB - Wavefronts reconstructed from measured gradients are composed of a straightforward integration of the measured data, plus a correction term that disappears when there are no measurement errors. For regions of any shape, this term is a solution of Poisson's equation with Dirichlet conditions (V = 0 on the boundaries). We show that for rectangular regions, the correct solution is not a periodic one, but one expressed with Fourier cosine series. The correct solution has a lower variance than the periodic Fourier transform solution. Similar formulas exist for a circular region with obscuration. We present a near-optimal solution that is much faster than fast-Fourier-transform methods. By use of diagonal multigrid methods, a single iteration brings the correction term to within a standard deviation of 0.08, two iterations, to within 0.0064, etc. PMID- 16477835 TI - Optical tomography of transparent objects with phase-shifting interferometry and stepwise-shifted Ronchi ruling. AB - An experimental setup for tomographic inspection of phase objects is presented. The system uses a common-path interferometer consisting of two windows in the input plane and a translating grating as its pupil. In the output, interference of the fields associated with replicated windows is achieved by a proper choice of the windows' spacing with respect to the grating period. With a rotating object in one window and a plane wave in the second one, the phase distribution of each projection is encoded as a corresponding digital image row, which, in turn, constructs a composite interferogram over the plane of a traditional sinogram. Phase stepping of composite interferograms can be achieved by a proper translation of the grating in order to obtain the unwrapped phase distribution as the corresponding sinogram. This sinogram allows tomographic reconstruction of phase slices by standard procedures. Composite interferograms and reconstructions for some transparent samples are shown. PMID- 16477836 TI - Diffraction of electromagnetic waves by periodic arrays of rectangular cylinders. AB - Reflection, transmission, and absorption of electromagnetic waves by periodic arrays of conducting or dielectric rectangular cylinders are studied by a finite difference time-domain technique. Truncated gratings made of lossless and lossy conducting and dielectric elements are considered. Results for surface current density, transmission, and reflection coefficients are calculated and compared with corresponding results in the literature, which are obtained by approximate or rigorous methods applicable only to idealized infinite models. An excellent agreement is observed in all cases, which demonstrates the accuracy and efficacy of our proposed analysis technique. Additionally, this numerical method easily analyzes practical gratings that contain a finite number of elements made of lossless, lossy, or even inhomogeneous materials. The results rapidly approach those for the idealized infinite arrays as the number of elements is increased. The method can also solve nested gratings, stacked gratings, and holographic gratings with little analytical or computational effort. PMID- 16477837 TI - Coherence solution for bidirectional reflectance distributions of surfaces with wavelength-scale statistics. AB - The scalar bidirectional reflectance distribution function (BRDF) due to a perfectly conducting surface with roughness and autocorrelation width comparable with the illumination wavelength is derived from coherence theory on the assumption of a random reflective phase screen and an expansion valid for large effective roughness. A general quadratic expansion of the two-dimensional isotropic surface autocorrelation function near the origin yields representative Cauchy and Gaussian BRDF solutions and an intermediate general solution as the sum of an incoherent component and a nonspecular coherent component proportional to an integral of the plasma dispersion function in the complex plane. Plots illustrate agreement of the derived general solution with original bistatic BRDF data due to a machined aluminum surface, and comparisons are drawn with previously published data in the examination of variations with incident angle, roughness, illumination wavelength, and autocorrelation coefficients in the bistatic and monostatic geometries. The general quadratic autocorrelation expansion provides a BRDF solution that smoothly interpolates between the well known results of the linear and parabolic approximations. PMID- 16477838 TI - Mode coupling approach to beam propagation in atmospheric turbulence. AB - A mode coupling approach based on the modal theory of coherence is suggested for the study of partially coherent beams in atmospheric turbulence. An approximate expression is derived for the mode power coupling coefficients, and some specific cases are studied using numerical methods. Several general results derived from the properties of the coupling coefficients are also presented. PMID- 16477839 TI - Exact solution to plane-wave scattering by an ideal "left-handed" wedge. AB - An exact analytical solution to the problem of plane-wave diffraction by a penetrable left-handed medium (LHM) epsilon = micro = -1 wedge of arbitrary angle (subject to valid physical constraints) is presented. Standard analysis involving discontinuous angular eigenfunctions and even/odd symmetry decomposition resulted in a discrete spectrum leading to a series solution resembling the traditional perfect electric conductor wedge solution but exhibiting the expected negative refraction phenomenology. Numerical results are presented, some of which seemed paradoxical but are explainable by classical means. A new type of illusory edge radiation is observed and explained. Also, a novel edge-launched interface standing wave is observed on the directly illuminated side. The exact analytical solution is verified by comparison with finite-difference time-domain simulation on causal LHM materials. PMID- 16477840 TI - Effective-medium theory for finite-size aggregates. AB - We propose an effective-medium theory for random aggregates of small spherical particles that accounts for the finite size of the embedding volume. The technique is based on the identification of the first two orders of the Born series within a finite volume for the coherent field and the effective field. Although the convergence of the Born series requires a finite volume, the effective constants that are derived through this identification are shown to admit of a large-scale limit. With this approach we recover successively, and in a simple manner, some classical homogenization formulas: the Maxwell Garnett mixing rule, the effective-field approximation, and a finite-size correction to the quasi-crystalline approximation (QCA). The last formula is shown to coincide with the usual low-frequency QCA in the limit of large volumes, while bringing substantial improvements when the dimension of the embedding medium is of the order of the probing wavelength. An application to composite spheres is discussed. PMID- 16477841 TI - Fast numerical method for electromagnetic scattering by rough layered interfaces: propagation-inside-layer expansion method. AB - Electromagnetic scattering from a stack of two one-dimensional rough surfaces separating homogeneous media is modeled with a rigorous integral formulation solved by the method of moments. We present an efficient numerical method for computing the field scattered by such rough layers, in reflection as well as in transmission. We call this method propagation-inside-layer expansion (PILE) due to its straightforward physical interpretation. To our knowledge, it is the first method able to handle problems for this configuration with a huge number of unknowns. We study the convergence of this method versus a coupling condition and validate it by comparison with results from the literature. PMID- 16477842 TI - Homogenization and scattering from heterogeneous media based on finite-difference time-domain Monte Carlo computations. AB - We present a method to study the scattering by heterogeneous media based on the two-dimensional (2D), finite-difference-time-domain method and a Monte Carlo algorithm. The inhomogeneities may reach wave-length size and their optical constants are in the visible and infrared domain. The algorithm is used to determine an effective propagation constant in a monodisperse medium from the observation of the energy decay in the medium. The result is compared over a large domain of volume fraction with the Keller and the Foldy-Twersky 2D models to determine the domain of their validity. Then the same approach is applied to homogenize the smallest particles in a bidisperse case and determine when such process is adequate. PMID- 16477843 TI - Analytical modeling of adaptive optics: foundations of the phase spatial power spectrum approach. AB - End-to-end simulation of adaptive optics (AO) systems allows high-fidelity modeling of system performance, but at the cost of long computation time. Analytical modeling, on the other hand, can provide much faster first-order performance estimates for a rapid exploration of the AO parameter space. In this paper, we present the foundations of a modeling method for the AO optical transfer function, based on an analytical description of the residual phase spatial power spectrum. The method has been implemented in an IDL-based code, PAOLA, and comparison with end-to-end simulations demonstrates the validity of the analytical approach. PMID- 16477844 TI - Gaussian beam weak scintillation: low-order turbulence effects and applicability of the Rytov method. AB - A generally applicable and computationally efficient description of random irradiance fluctuations induced by single scattering from distributed low-order turbulence (LOT) phase fluctuations is developed for Gaussian beams in the weak scintillation regime. The LOT solution describes irradiance statistics resulting from coarse beam irradiance fluctuations such as beam wander and beam breathing and will generally underestimate the true scintillation owing to the neglect of higher orders. For a subset of beam and turbulence settings that naturally result in non-log-normal irradiance behavior in the weak regime, the LOT solution closely approaches the exact solution and accurately describes the irradiance statistics for any point on the observation plane. For the same settings, beam wave scintillation theory derived from the Rytov perturbation method yields inaccurate predictions owing to an inherent confinement to log-normal behavior. Examples that naturally exhibit non-log-normal irradiance behavior include focused beams on horizontal paths and collimated beams on ground-to-space paths. The complementary nature of the two scintillation theories (LOT and Rytov) enables a hybrid combination that yields accurate and convenient scintillation predictions for any case exhibiting weak scintillation regardless of irradiance behavior. Comparison of hybrid model predictions with wave optics simulation data reveals excellent agreement. PMID- 16477845 TI - Sky coverage estimates for adaptive optics systems from computations in Zernike space. AB - A sky coverage model for laser guide star adaptive optics systems is proposed. The atmosphere is considered to consist of a finite number of phase screens, which are defined by Zernike basis polynomials, located at different altitudes. These phase screens are transformed to the aperture plane, where they are converted to laser and natural guide star wavefront sensing measurements. These transformations incorporate the cone effect due to guide stars at finite heights, anisoplanatism due to guide stars off axis with respect to the science object, and adaptive optics systems with multiple guide stars. The wavefront error is calculated tomographically with minimum variance estimators derived from the transformation matrices and the known statistical properties of the atmosphere. This sky coverage model provides fast Monte Carlo simulations over random natural guide star configurations, irrespective of telescope diameter. The Monte Carlo simulations outlined show that inclusion of a finite outer scale for the atmosphere significantly reduces the median wavefront error, that increasing the number of laser guide stars in the asterism reduces the median wavefront error, and that a larger natural guide star patrol field provides a smaller median wavefront error when there is a low star density in the field. PMID- 16477846 TI - Irradiance formations in hollow straight light pipes with square and circular shapes. AB - We investigated, both analytically and numerically, the irradiance formation of an asymmetrically located Lambertian light source in hollow straight light pipes with square and circular shapes. The uniform irradiance distribution in a square light pipe and hot-spot localization in a circular light pipe were examined and determined semianalytically. Typical factors of influence, such as light-pipe length, width, and source size, were identified with extensive simulation. When the ratio of light-pipe length and width was less than 0.5, the deviation from uniformity could be more than 20%. But once the source size was large enough (approximately half of the incident port), such that the Lambertian characteristics of the source dominated the irradiance distribution, the uniformity deviation was reduced. Furthermore, a quantity of root-mean-square circular differences was defined in order to identify the shape deformation of the light pipe; it was found that the peak value of the hot spot decreased exponentially with the deformation scale. The influence of nonperfect reflectivity of the pipe wall on irradiance formation was also examined for a square light pipe; when the reflectivity is larger than 90%, the difference in uniformity is less than 10% and uniform irradiance remains, provided that the ratio of light-pipe length and width is larger than 1; even the source is located asymmetrically. PMID- 16477847 TI - Carrier phase component removal: a generalized least-squares approach. AB - In fringe projection profilometry, an object shape is evaluated through phase distribution extracted from a projected fringe pattern. For parallel illumination geometry, the carrier phase component introduced by the fringes is spatially linear, whereas nonparallel illumination would lead to a nonlinear carrier. In this study, a general approach for the removal of a nonlinear-carrier phase component is proposed. A series expansion technique is used to approximate the carrier phase function, and a least-squares method is developed to estimate the unknown coefficients of the series. The theoretical analysis is given on the basis of a divergent illumination geometry with carrier fringes in the x direction. The method is also extended to include a curved surface-fitting approach, which is applicable to various measurement system geometries. PMID- 16477848 TI - Determination of the axial velocity component by a laser-Doppler velocity profile sensor. AB - We report about the determination of the axial velocity component by a laser Doppler velocity profile sensor that is based on two superposed fanlike interference fringe systems. Evaluation of the ratio of the Doppler frequencies obtained from each fringe system yields the lateral velocity component and the axial position inside the fringe system. Inclined particle trajectories result in chirped burst signals, where the change of the Doppler frequency in one burst signal is directly related to the axial velocity component. For one single tracer particle it is possible to determine (i) the lateral velocity component, (ii) the axial velocity component including the direction, and (iii) the axial position of the tracer trajectory. In this paper we present the measurement principle and report about results from simulation and experiments. An uncertainty of the axial velocity component of about 3% and a spatial resolution in the micrometer range were achieved. Possible applications of the sensor lie in three-component velocity measurements of flow fields where only one optical access is available. PMID- 16477849 TI - Operator expansions for polarization mode dispersion analysis and compensation. AB - We present analytic third- and fourth-order expansions of the Jones matrix as products of exponentials of individual matrices. In our first formalism, these are polarization mode dispersion (PMD) matrices of definite orders. We then discuss an alternative procedure that instead employs exponentials of general skew-Hermitian matrices with a low-order dependence on the deviation of the optical frequency from a central reference frequency. Our expressions correspond to PMD compensators formed from a succession of relatively simple optical components each of which has the frequency response of a single operator in the product. PMID- 16477850 TI - Integral equation analysis of scattering by a spherical microparticle coupled to a subwavelength-diameter wire waveguide. AB - A rigorous integral equation analysis of the coupling between a fiber waveguide and an adjacent spherical particle is developed. The solution is obtained by applying the entire-domain Galerkin technique, based on Mie-type spherical wave expansion of the field in the sphere and the use of dyadic Green's function of the fiber waveguide. The conversion between cylindrical and spherical wave functions is done through classic analytical formulas. The analysis is applied to numerically investigate transmission through silica wires of subwavelength diameter in the presence of particles of comparable size. The results show the possibility of sensing microparticles through the reduction of transmitted power, which is maximum for certain critical values of the fiber diameter. PMID- 16477851 TI - Pseudodielectric functions of uniaxial materials in certain symmetry directions. AB - The pseudodielectric function is often used to represent ellipsometric data and corresponds to the actual dielectric functions of materials when there is no surface overlayer and the material is isotropic. If a uniaxial material is oriented such that the optic axis is in the plane of incidence or is perpendicular to the plane of incidence, then the cross-polarization terms are zero and appropriate pseudodielectric functions can be determined from the ellipsometry data. We calculate the pseudodielectric functions for uniaxial crystals in three primary symmetry directions: (1) the optic axis is perpendicular to the plane of incidence, (2) the optic axis is in the plane of the sample surface and parallel to the plane of incidence, and (3) the optic axis is in the plane of the sample surface and perpendicular to the plane of incidence. These results are expanded in terms of the difference in the ordinary and extraordinary dielectric functions and compared with the approximation of Aspnes [J. Opt. Soc. Am. 70, 1275 (1980)]. Comparisons are made with experimental results on oriented crystals of rutile (TiO2), and a simple procedure is presented to determine the complex dielectric function from standard ellipsometry techniques. PMID- 16477852 TI - Geometric phase lens. AB - The design of a lens that modulates the geometric phase of an optical beam by manipulating its polarization is presented. To produce such a geometric phase element with a spatially varying phase function, one needs a wave plate with varying orientation. One can use subwavelength grooves to produce form birefringence, but the variation in orientation generally leads to branch points in the groove pattern. These branch points do not affect the phase of the traversing beam directly because the grooves are subwavelength. However, they do produce errors in the groove orientation, which indirectly leads to errors in the geometric phase function that is implemented. A design procedure is provided to compute the groove pattern for such a rotationally symmetric geometric phase element; and, with the aid of a numerical simulation, the effect of the branch points in the groove pattern on its performance is investigated. PMID- 16477853 TI - Computational studies of optical textures of twist disclination loops in liquid crystal films by using the finite-difference time-domain method. AB - Optical images of textured liquid-crystal films containing various types of twist disclination loops are computed using an approximate matrix method and a direct numerical simulation based on the finite-difference time-domain (FDTD) method. The selected defects introduce large multidirectional spatial gradients in the optic axis, mimicking the orientation textures that arise in the construction and use of biosensors based on liquid-crystal vision. It is shown that under these experimentally relevant conditions, the matrix method fails to capture important signatures in the transmitted light intensity under crossed polarizers. The differences between the predictions by the two methods are analyzed with respect to gradients in the optic axis. We show that the FDTD method is a useful tool to perform computational optics of textured liquid-crystal films. PMID- 16477854 TI - Ill-posed inverse problem in diffraction optics. tolerance analysis of diffractive lenses and gratings. AB - A method is proposed for the prediction of deviations of the phase polynomial coefficients of diffractive optical elements from their nominal values due to fabrication errors. This problem represents an ill-posed inverse problem in diffraction optics. Its solution is based on the regularization technique resulting in an approximate stable (rather than exact but unstable) solution of the inverse problem. The proposed method can become a useful tool, providing optical designers with a possibility of including these deviations into the tolerance analysis of optical devices. PMID- 16477855 TI - Arterial injuries in civilian practice in Lagos, Nigeria. AB - OBJECTIVE: This is a retrospective study of patients managed at the Lagos University Teaching Hospital for peripheral arterial injuries from January 1995 to April 2003. The aim was to study the pattern of peripheral arterial injuries in Lagos. Nigeria and to look at the outcome of management and see what improvements could be made in future. METHOD: Data was collated from case notes of patients, operation register in theatre and admission and discharge books from the surgical wards and the data bank of consultants involved in patients' management. RESULT: Forty-one patients. 37 males and 4 females. were treated within the study period (M:F ratio of 9:1). Twenty three patients (56.1%) suffered gunshot injuries during armed robbery attacks while 9 patients (22.0%) had stab injuries in civilian violence. Twenty-one patients (3 with gunshot injuries and 18 non gunshot penetrating injuries) were managed by direct suturing of vessels. Eight patients had prosthetic graft interposition while 2 patients had reversed saphenous vein grafts. Two patients had the superficial branches of their radial arteries tied up at the wrist. CONCLUSION: Gun shot injuries from armed robbery attack was the commonest cause of peripheral arterial injuries in this environment during the period of study. PMID- 16477856 TI - Genitourinary tuberculosis in Nigeria; a review of thirty-one cases. AB - OBJECTIVE: To report and discuss the characteristics of genitourinary tuberculosis as observed in a sub-Saharan African setting, where reports suggest its rarity despite reported high incidence and prevalence of pulmonary tuberculosis. PATIENTS AND METHOD: Thirty one consecutive patients with discharge diagnoses of genitourinary tuberculosis seen in a small community-based hospital over a five-year period were retrospectively studied. RESULT: There were 28 males and three females, with a mean age of 37.6 (SD 11.5) years. Forty- three sites were involved in seven organs: epididymis 25 (58.1%), kidney 7 (16.3%), testis 4 (9.3%), bladder 3 (7.0%), ureter 2 (4.7%), prostate 1 (2.3%), and cord of the testis 1 (2.3%). The commonest presenting features were scrotal/testicular mass with or without pain/tenderness (80.6%), fever/headache (51.6%), and dysuria (22.6%). Other common features were back, loin, or abdominal pain/tenderness. hydrocoele, scrotal abscess, and haematuria. 26.9% had evidence of concurrent or previous pulmonary tuberculosis, and 9.1% were positive on HIV 1 & 11 screening. CONCLUSION: With the prevailing conditions in sub-Saharan Africa and most of the developing world and the slightly different characteristics of the disease in our environment, diagnosis of genito-urinary tuberculosis may be difficult. It is advised that patients with unexplained symptoms in the urinary tract should be investigated for tuberculosis. PMID- 16477857 TI - Health care workers' knowledge on HIV and AIDS: universal precautions and attitude towards PLWHA in Benin-City, Nigeria. AB - OBJECTIVE: Health care workers are at risk of becoming infected with blood-borne pathogens, including HIV. The study was designed to test health care workers knowledge about HIV transmission, universal precautions and their attitude towards people living with HIV and AIDS. DESIGN: A cross-sectional study. SETTING: University of Benin Teaching Hospital, Benin-City, Nigeria. PARTICIPANTS: 120 Health Care Workers (HCWs) who were occupationally exposed to patient's blood and body fluids completed a self administered structured questionnaire between March and May 2004. The HCWs consisted of 50 doctors drawn from obstetrics and gynaecology (25) and surgery departments (25). 70 nurses from accident and emergency unit (23), labour ward (18), labour ward theatre (4), main surgical theatre (22) and family planning clinic (3). RESULTS: The mean age of the health care workers and duration of practice were 39.8 +/- 8.0 years and 14.0 +/- 8.2 years respectively. Though many of the respondents demonstrated good knowledge about HIV transmission, more than 25% of them thought that HIV could be transmitted through saliva, vomit, faeces and urine. They over estimated their risk of acquiring HIV infection following needle stick injury, exposure of mucocutaneous membrane and intact skin to infected blood and body fluids. There was poor adherence to universal precautions which was attributed to lack of knowledge and availability of materials in 48% and 60% of the workers respectively. Over 40% of the health care workers exhibited discriminatory attitude towards people living with HIV and AIDS. There was no statistical significant difference (p > 0.05) in the knowledge of HIV and AIDS transmission and infection prevention practices amongst the doctors and nurses. Similarly there was no significant difference in their discriminatory attitude towards PLWHA. CONCLUSION: We recommend that seminars, workshops should be organized on a continuous basis for health care workers on universal precautions, stigma and discrimination reduction. Those trained should train others on the job. The institution should also make available materials needed to protect workers against the risk of acquiring pathogenic infection in the course of providing health services to their patients. PMID- 16477858 TI - Prevalence of dermatophytosis in University of Nigeria Teaching Hospital, Enugu, Nigeria: any change in pattern? AB - A total of 344 specimens of skin, hair and nail clippings, were examined for the presence of dermatophytes during the period May 2003 - April 2004 at the UNTH.Enugu. Out of these, 65 were found positive for dermatophytes. T. soudanense was the most frequently isolated species representing 70.8% of isolates, followed by T. mentagrophytes 12.3%. Adults were more predominantly infected than children. 89.2% and 10.8% respectively. T. soudanense was the only dermatophyte that was recovered from all sites apart from the buttocks. Compared to an earlier study in 1975, there was a significant decrease in the prevalence of T. capitis (P < 0.05) and this explains the decrease in the prevalence of M. andounii, as this dermatophyte is the most common cause of T. capitis in Enugu. Nigeria. An improvement of Personal hygiene, better living conditions and improved socio economic conditions has led to a decrease of T. Capitis, as well as general awareness by the populace. PMID- 16477859 TI - A comparison of sphygmomanometric and oscillometric methods of blood pressure measurements in adult in-patients. AB - The study compared two non-invasive methods of blood pressure measurements used in the anaesthetic management of patients, the conventional sphygmomanometric and the oscillometric methods. One hundred adult in-patients were involved, and 400 blood pressure measurements were done with the two devices. The sphygmomanometric measurements were higher in systolic, diastolic and mean arterial pressure values. Correlation coefficient values between the two methods were r = 0.97, 0.81, 0.95 for systolic, diastolic and mean arterial pressures respectively, (p < 0.05). We conclude that there is a statistically significant difference in haemodynamic values determined by the two devices. This difference is however, not clinically significant to warrant a recommendation of adjustment when comparing values determined by the two devices. PMID- 16477860 TI - Antimicrobial susceptibility pattern of Klebsiella species from Ebonyi State University Teaching Hospital Abakaliki, Nigeria. AB - Klesiella specie isolated from clinical specimens from Ebonyi State University Teaching Hospital (EBSUTH). Abakakliki were studied to determine the antimicrobial susceptibility pattern. Between January, 2003 and September 2004 a total of 3.600 specimens processed in the routine Medical Microbiology laboratory of EBSUTH, of which 245(6.8%) yielded Klebsiella species, with 84 from out - patients and 161 from in - patients. The number of isolates from various samples were: Urine 126, Sputum 37 Endocervical swab 13, Aspirates 8, High Vaginal Swab 7, Blood 3, Eye Swab, Ear Swab and Cerebrospinal fluid were 2 samples each. Organisms were identified by conventional methods. Antimicrobial susceptibility was done by the disk diffusion methods. The antimicrobial disk used include: Ceftazidime, Cefuroxime, Cefotaxine, Augmentin, Pefloxacin (30ug), Doxycyline (25ug) Genticin (10 ug) Ciprofloacin and Ofloxacin (5ug) each and Erythromycin (15ug). All were Oxoid products. Results were interpreted according to NCCLS criteria. Klebsilla species were isolated mostly from urine specimens (51.4%) followed by wound swabs (18.4%). Antimicrobial susceptibility to various groups drugs used was generally poor. The most sensitive antimicrobial was Ciprofloxacin with 121(49.4%) isolates susceptible to it, followed by Gentamicin with 95 (38.8%) and Ceftazidime with 90(36.7%). Seventeen isolates were multiresistant to all the antimicrobial agents used. The result of this study will help in the empiric therapy of infection caused by Klebsiella species in Ebonyi State University Teaching Hospital, Abakaliki, Nigeria but continuous surverillance of antimicrobial resistance of the organnisn is very necessary in the formulation of a sound antibiotic policy in the hospital. PMID- 16477861 TI - The determinants of seizure severity in Nigerian epileptics. AB - BACKGROUND: Epilepsy is a chronic disease and the control of seizures is central to its management. While seizure frequency has been the traditional index of epilepsy control. severity of seizures is probably as important as seizure frequency in this regard. Seizure severity scales have therefore been developed to assess the impact of antiepileptic drugs on seizure control. The eight items of the national hospital seizure severity scale were applied in this study to Nigerian subjects with epilepsy to determine which aspects of seizure severity were considered relevant from the patients' perspective. MATERIALS AND METHODS: Twenty-eight confirmed subjects with epilepsy at the University College Hospital, Ibadan, were studied. The National Hospital seizure severity scale questionnaire was administered to all subjects. This assesses generalisation of seizures, falls, injuries, urinary incontinence, warning interval before loss of consciousness, automatisms and time of recovery on a graded scale. RESULTS: The most frequent indices of seizure severity in Nigerian epileptics is the generalisation of seizures in 85.7% of subjects, incontinence of urine in 78.6%, absence of protective warning time in 75% and occurrence of serious injuries in 71.5%. Occurrence of falls and disruptive automatisms were less frequent. Seizure severity scores were worse in subjects with longer duration of epilepsy and in those on monotherapy. CONCLUSIONS: The indices of seizure severity that occurred most frequently in Nigerian subjects with epilepsy were generalisation of seizures, incontinence of urine, absence of protective warning time and serious injuries. These may need to be addressed in the management of epilepsy patients. Subjects on monotherapy in this study had worse seizure severity scores and this may indicate the need to consider early rational polytherapy in order to improve seizure control. PMID- 16477862 TI - Knowledge and perceptions of HIV/AIDS and mother to child transmission among antenatal mothers at Nnamdi Azikiwe University Teaching hospital, Nnewi. AB - OBJECTIVE: Knowledge of HIV/AIDS by pregnant mothers is very important in the prevention of mother to child transmission. This study evaluates the knowledge and perceptions of HIV/AIDS and mother to child transmission among pregnant women attending antenatal clinic at a University Teaching Hospital. METHODOLOGY: Pre tested questionnaires were interviewer administered to 312 pregnant women randomly selected at the antenatal clinic of the Nnamdi Azikiwe University Teaching Hospital Nnewi. RESULTS: The level of awareness of HIV/AIDS among antenatal mothers was very high (99%) and the main sources of information were radio (44.7%), television (38.8%), and print media (34.0%). Though majority (94.2%) was aware HIV infection can coexist with pregnancy, only 76.9% were aware of mother to child transmission. Transplacental (46.1%), breastfeeding (31.7%), and vaginal delivery (16.3%) were the commonly identified routes of vertical transmission. Surprisingly, eighteen respondents (5.8%) indicated that caesarean section is a possible route of vertical transmission. CONCLUSION: Though the percentage of HIV/AIDS knowledge is high, the level of knowledge and perceptions of mother to child transmission is inadequate. This suggests the need to scale up health education about mother to child transmission in our health facilities. PMID- 16477863 TI - Knowledge, attitude and practice of aspects of laboratory safety in Pathology Laboratories at the University of Port Harcourt Teaching Hospital, Nigeria. AB - OBJECTIVE: To assess current knowledge, attitudes and practice of aspects of laboratory safety in pathology laboratories at the University of Port Harcourt Teaching Hospital in view of perceived inadequacies in safety practices in clinical laboratories in developing countries. MATERIALS AND METHODS: Sixty (60) self- administered questionnaires were distributed to all cadres of staff in four (4) different laboratories (Chemical Pathology, Haematology, Blood bank and Medical Microbiology) at the Hospital. RESULT: Gross deficiencies were found in the knowledge, attitudes and practice of laboratory safety by laboratory staff in areas of use of personal protective equipment, specimen collection and processing, centrifuge--related hazards, infective hazards waste disposal and provision and use of First Aid Kits. CONCLUSION: Issues pertaining to laboratory safety are not yet given adequate attention by both employers and employees in developing countries in this ear of resurgence of diseases such as HIV/AIDS and Hepatitis Band C, is emphasized. PMID- 16477864 TI - Increasing incidence of bacterial, resistance to antibiotics by isolates from the urinary tract. AB - This is a prospective study to determine the prevalence of antibiotic resistance among organisms causing urinary tract infections in a Teaching Hospital between August 2003 and July 2004 and to compare them with an earlier study in 1993. A total of 1,104 urine samples were collected in sterile universal containers from patients attending University of Nigeria Teaching Hospital and studied. All samples showing significant bacteriuna were studied and isolates identified using standard bacteriogical methods. Antibiotic sensitivity testing was performed on sensitivity test agar (Biotec, UK) using the disc diffusion method in accordance with the National Committee for Clinical Laboratory Standards (5). The results were compared with a previous study in 1993. A total of 3.36 urinary isolates were identified, with the coliforms being the most predominant (51.2%). followed by Staphylococcus aureus and Proteus species (28.9% and 7.1%) respectively. Compared to the earlier study, a significant increase in the resistance of the urinary pathogens to ofloxacin, ciprofloxacin and nalidixic acid was observed, however there was a decrease in the resistance to nitrofurantoin (p < 0.05) using chi-square test. The results of this study should now alert doctors about the increasing possibility of treatment failures, when ciprofloxacin, ofloxacin and nalidixic acid are used for the treatment of urinary tract infections without laboratory testing. A multi-faceted approach including continued and improved surveillance, a reduction in the unnecessary use of antibiotics and infection control are necessary. PMID- 16477865 TI - Histological review of melanocarcinoma in Port Harcourt. AB - BACKGROUND: Melanocarcinoma is a malignancy of melanocytes affecting more females; and commoner in the farming population. It is also associated with high morbidity and mortality. OBJECTIVE: To determine the pattern, outcome of melanocarcinoma and a survey of presentation of the tumor in Port Harcourt was undertaken. DESIGN: A retrospective descriptive study for 11 years (1st January 1990- 31st December 2000). SETTING: University of the Port Harcourt Teaching Hospital, Port Harcourt. METHOD: Histological slides previously stained with hematoxylin and eosin of 15 cases of melanocarcinoma were retrieved and re- evaluated for the study. Special stain like Mason Fontana stain was also used for proper diagnosis of some of the tumors. Tissue slides or blocks which could not be located and those with inadequate documentations were excluded from the study. The 15 cases were staged according to Clark's Staging System. RESULT: Melanocarcinoma is rare in this environment as it accounted for 0.8% of total malignancies for the period under review. The age ranged from 39-76 years. Majority of the cases were female with F:M ratio of 3:2. The peak (73.3%) frequency of occurrence was among the age group (51-70) years. Only the nodular and the acral leniginous types were seen, of which the nodular type was the commonest (80%). The feet and the legs were the commonest predilection sites (46.7%) while the head and neck as well as the knee were the least site of occurrence. There was no stage I and II melanocarcinoma in this study. The cancers were seen in stage V, IV and III in decreasing frequency. CONCLUSION: melanocarcinoma. thought rare in this environment, present at advanced stage of the disease. Public enlightenment and awareness campaign as to the physical characteristics of the tumor is needed to make patients present early for diagnosis of the disease and treatment. PMID- 16477866 TI - Facial neuralgias: analysis of the different types seen at Lagos University Teaching Hospital, (Luth). AB - OBJECTIVE: To highlight the presentations, characteristics. the difficulties in diagnosis, treatment and response to treatment types of facial neuralgias seen at Lagos University Teaching Hospital. METHODS: Twelve patients with facial neuralgias diagnosed and treated in dental clinic of the Lagos University Teaching Hospital were studies. Using strict for diagnosis, patients were categorized into: trigeminal, glosspharyngeal and post herpetic neuralgias. RESULTS: Eight patients had trigeminal neuralgia; three patients had post herpetic neuralgia and one patient had glossopharyeal neuralgia. In six patients with Trigeminal neuralgia. mandibular branch was affected, while in the two patients maxillary branch was affected. Six patients with Trigeminal neuralgia responded to carbamazepine alone and 2 had additional drugs. The only patients with glosspharyngeal neuralgia responded to carbamazepine. One patient with post herpetic neuralgia tested positive for HIV. All the post herpetic neuralgia responded poorly to carbamezepine. CONCLUSION: Facial neuralgias are uncommon and usually present in the dental clinic. They can easily be misdiagnosed with resulting inappropriate. Correct diagnosis and treatment with carbamezepine is beneficial in majority of patients. PMID- 16477867 TI - Tuberculosis: current trends in diagnosis and treatment. AB - Among communicable diseases, tuberculosis (TB) is the second leading cause of death worldwide, killing nearly 2 million people each year. It is estimated that about one-third of the world population are infected with TB (2 billion people) and about 10% of this figure will progress to disease state. Most cases are in the less-developed countries of the world. Tuberculosis incidence has been on the increase in Africa, mainly as a result of the burden of HIV infection. Definitive diagnosis of tuberculosis remains based on culture for Mycobacterium tuberculosis, but rapid diagnosis of infectious tuberculosis by simple sputum smear for acid fast bacilli remains an important tool, as more rapid molecular techniques are being developed. Treatment with several drugs for 6 months or more can cure more than 95% of patients. Direct observation of treatment, a component of the recommended five-element DOTS strategy, is judged to be the standard of care by most authorities. Currently only a third of cases worldwide are treated using this approach. There may be need to modify the treatment modalities especially with the choice of drugs and duration of therapy when TB infection occurs in special situation like pregnancy, liver disease, renal failure or even in coexistence with HlV/AIDS or the drug resistant state. PMID- 16477868 TI - Prosthetic management of an epileptic patient. AB - This case report illustrates the problems of tooth loss in an epileptic patient. The patient presented with a broken denture following a seizure. She gave a history of breaking and swallowing her dentures during seizures. Before presentation she had worn five upper removable partial dentures. An upper removable partial denture with increased thickness of the acrylic palatal was fabricated and fitted satisfactorily. The patient was taught how to insert and remove the prosthesis as quickly as possible. Epileptic patients can use dentures but run the risk of frequently breaking and swallowing them during seizures. The risk can be reduced if patients and relatives are taught how to remove the dentures prior to or during seizures. PMID- 16477869 TI - Heterotopic pregnancy in a natural conception cycle. A case report. AB - We report a case of combined intrauterine and tubal pregnancy in a 32 year old para. The patient presented at the Gynaecological emergency unit, with lower abdominal pain, and vaginal bleeding. A pelvic scan confirmed an intrauterine pregnancy. While being managed as a case of threatened abortion, her condition deteriorated and the likelihood of an ectopic pregnancy was entertained. A repeat scan revealed significant peritoneal collection. suggestive of haemoperitoneum, and laparotomy confirmed heterotopic pregnancy. She had a right salpingectomy without complications. The patient had a supervised antenatal care and safe delivery of the intrauterine pregnancy. PMID- 16477870 TI - An accessory flexor of the fifth toe. AB - The presence of accessory muscles and other organs on the lower limbs of some individuals have variously been reported in the literature. We report an unusual muscle located on the plantar surface of the left foot of a cadaver, which had not been previously described. This muscle originated from the tendon of tibialis posterior and inserted into the middle phalanx of the fifth toe. It differed from the 'expansions' of the tibialis posterior tendon, which usually pass from its insertion on the navicular bone to other tarsal bones and are ligamentous in nature. The muscle produced flexion of the fifth toe and is innervated by the medial plantar nerve. Awareness of this is important to Anatomists and surgeons, especially those working on the foot. PMID- 16477871 TI - Familial thyrotoxicocis in five Nigerians. AB - We describe the occurrence of thyrotoxicosis in four Nigerian families. Hitherto. the descriptions of familial thyrotoxicosis have been confined to the Caucasian population and only recently in a Chinese family. This is the first description of familial thyrotoxicosis in the Nigerian population. The mutation analysis of the genomic DNA of the TSH receptor of these patients is required to define the genetic mutations that caused the disease. We recommend that a high index of suspicion for familial thyrotoxicosis should be exercised in the clinical evaluation of patients who present with hyperthyroidism. PMID- 16477872 TI - Prevalence of gynaecological diseases in Nnewi, Nigeria. PMID- 16477873 TI - Histological examination in assessment of ultraviolet-induced suppression of contact hypersensitivity response. AB - The evaluation of contact hypersensitivity (CHS) reaction is one of the methods used in the assessment of the immune status of an organism after UV radiation. The aim of the study was to compare usefulness of visual scoring system and histological morphometry in the assessment of CHS response after exposure of humans to solar simulated radiation (SSR). The study included 140 healthy volunteers, 33 people were irradiated for 2 days, 34 - for 10 days and 33 - for 30 days with SSR. Forty non-irradiated individuals served as controls. All the volunteers were sensitized with diphenylocyclopropenone (DPCP) 24 h after final exposure. Statistical analysis comparing intensity of CHS reaction based on visual score between irradiated groups and non-irradiated group revealed no differences (p>0.05). We found a significant difference in epidermal thickness between healthy skin and irradiated groups (p<0.05) and a positive correlation between intensity of spongiosis and clinical score for CHS response at 3.2 DPCP site (p<0.000001). A negative correlation between time of irradiation and spongiosis score was revealed (R=-0.28; p<0.001). We conclude that histological examination of biopsies taken from one of the series of elicitation sites is a reliable and sensitive method in the evaluation of CHS response after UVR. PMID- 16477874 TI - Germline BRCA1 mutations and G/C polymorphism in the 5'-untranslated region of the RAD51 gene in Polish women with breast cancer. AB - Breast cancer is a significant cause of morbidity and mortality in Western countries. In the present work the distribution of genotypes and frequency of alleles of the RAD51 G/C polymorphism and the frequency of BRCA1 germ-line mutations in patients with breast cancer were investigated. One hundred breast cancer women provided blood for mutation analysis. Blood samples from age matched healthy individuals (n=106) served as control. The G/C polymorphism and BRCA1 mutations were determined by PCR-RFLP methods. The distribution of the genotypes of the G/C polymorphism RAD51 in both control and patients did not differ significantly (P>0.05) from those predicted by the Hardy-Weinberg distribution. There were no significant differences in the genotype distributions and allele frequencies between node-positive and node-negative patients. In the present study one Ex20insC mutation of BRCA1 gene was identified in women with breast cancer. Our study implies that the G/C polymorphism of the RAD51 gene may not be directly involved in the development and/or progression of breast cancer. The lack of detectable BRCA1 germ-line mutations in most cases suggests that there are probably additional, as yet unidentified genes predisposing to this disease. PMID- 16477875 TI - Why do we go on screening? AB - As healthcare rules are becoming better organized and as vaccination against Human Papillomavirus (HPV) infections is arising, the conditions of cytological screening for cervical carcinoma may have to be modified in the near future. After reminding the Quality Control constraints of the Pap test and the technical and epidemiological conditions for the HPV detection as a precursor of cervical carcinoma, the present paper wishes to discuss the possible role of vaccine on the screening strategy. PMID- 16477876 TI - DNA ploidy, cyclin D1, bcl-2 and lymphocytic infiltration of the tumor microenvironment as prognostic factors in laryngeal cancer patients. AB - The aim of this study was to evaluate correlations between DNA ploidy type, the immunostaining of cyclin D1, bcl-2 and the intensity of lymphocytic infiltration of the tumor microenvironment in relation to the histopathological G differentiation and pTNM classification. Thirty two patients were treated surgically for laryngeal cancer with total or partial laryngectomy in the Department of Otolaryngology Zabrze. The percentage of bcl-2 immunostaining was showed in 53% of the cases and was found to correlate with G differentiation and the patients' age. Cyclin D1 antigen stained positively in 24 cases (75%). Expression of cyclin D1 correlated with cancer stage. Cyclin D1 negative stain was found in T4-stage. DNA ploidy was examined in 19 cases. Aneuploidy was found in 5 cases only, while the rest were diploid. DNA ploidy value correlated with cyclin D1 expression. All paraffin sections were found to contain lymphocytic infiltrations of CD43 and CD45RO phenotype in the tumor front. Some cases showed high intensity of lymphocytic infiltrations of CD45RO phenotype. The intensity of CD43 lymphocytic infiltrations in the tumor front was related to the expression of cyclin D1 and bcl-2. PMID- 16477877 TI - Immunohistochemical expression of bcl-2 in Dukes' stage B and C colorectal carcinoma patients: correlation with p53 and ki-67 in evaluating prognostic significance. AB - The objective of this study was to evaluate the expression of bcl-2 in Dukes' stage B and stage C (AJCC/UICC stage I and III) colorectal adenocarcinomas and to examine its association with clinicopathological features, p53, ki-67 and long term outcome. Paraffin embedded specimens from 61 patients with Dukes' stage B (AJCC/UICC stage I) and 39 patients with Dukes' stage C (AJCC/UICC stage III) colorectal adenocarcinoma who were treated with surgery were assessed. We used immunohistochemistry to determine the expression of bcl-2, p53 and ki-67 with a five-year follow-up. Positive bcl-2 expression was seen in 27 cases (27%). Expression of bcl-2 protein was related to tumor stage (p=0.0117). There was very strong evidence of an association between bcl-2 staining and ki-67 score (p<0.001). There was a trend towards increased survival in patients whose tumors expressed bcl-2 protein (p=0.001). When entered into a multivariate analysis model, which also included p53 and stage, bcl-2 staining emerged as a prognostic indicator variable. Expression of bcl-2 appears to be useful in selecting a group of colorectal cancer patients with a better prognosis. PMID- 16477878 TI - Adequacy of trephine bone marrow biopsies: the doctor and the patient make a difference. AB - Reports of 1938 trephines submitted from five institutions over a 30-month period were analyzed looking for associations between the hospital of origin, operator, bone marrow pathology, patient's age and the biopsy quality. The arbitrary adequacy criteria (min 10 mm of interpretable marrow or min 10 intertrabecular spaces) were fulfilled by 61.9% of the biopsies. The performance of individual operators varied from 15.9% to 87.8% of adequate trephines. The group of doctors performing more than 100 biopsies in the study period had satisfactory results. The intermediate group (20-100 biopsies) was the least homogenous, and on the average had the poorest biopsy quality. The biopsy quality was influenced by diagnostic categories, correlated positively with bone marrow fibrosis and negatively with the patient's age. The trephine quality in practice may be lower than the published or declared standards. Ideally the procedure should be executed by the practitioners making more than one trephine a week. Prior to the biopsy it is possible to estimate the level of difficulty posed by an individual patient and use this information to minimize the risk of obtaining an inadequate core. PMID- 16477879 TI - Primary amyloid tumors of the lungs--six cases. AB - Over a period of nine years, the authors followed up six cases of primary amyloid tumors of the lungs in patients at the mean age of 58.5 years. All the patients were suspected of bronchial carcinoma and they were subjected to surgical treatment. The duration of follow-up varied, but their postoperative status was satisfactory. Immunohistochemical reactions showed deposits of AL amyloid in five cases; in one of these patients, pulmonary amyloid tumors were related to marginal zone lymphoma of the lung. In one case, the accumulated amyloid was transthyretin. PMID- 16477880 TI - Distribution of substance P nerve fibers in osteoarthritis knee joint. AB - The objective of the study was to evaluate the distribution of substance P immunoreactive nerve fibers in osteoarthritis knee and to determine whether a degenerative disease has any influence on the occurrence of neuropeptide containing fibers, positively stained for substance P. Twenty consecutive patients, 16 females and 4 males, with gonarthrosis participated in the study. For comparison we used the group of 20 patients, 14 females and 6 males, operated on because of traumatic lesion of the knee. The medial and lateral retinaculum, medial compartment synovium and infrapatellar fat pad of these two groups of patients were evaluated using monoclonal antibody to substance P (PEPA40, Serotec Ltd, UK). The slices were examined semi-quantitatively for nerve fibers showing substance P expression. The values were analyzed with ONE WAY ANOVA test, which was then corrected with LSD test, at the level of significance p<0.05. There were no statistical differences in distribution of substance P nerve fibers in the fat pad, lateral and medial retinaculum or synovium between both groups, as well as in the each study group (p<0.05). The results allow us to speculate that different biomechanical axial disturbances of the knee could have the same influence on substance P-positive mechanoreceptors of the soft tissues around the joint modulating the pain pathway in knee osteoarthrosis. PMID- 16477881 TI - Primary intrathoracic biphasic synovial sarcoma--a case report. AB - The authors present a rare case of a synovial sarcoma involving both pleural cavities in a 66-year old woman, confirmed by the t(X;18) translocation detected using the FISH method. PMID- 16477882 TI - Jellyfish species distinction has treatment implications. PMID- 16477883 TI - Dysmorphic findings in persons with fragile X syndrome. PMID- 16477884 TI - Medicare Part D: practical and policy implications for family physicians. PMID- 16477885 TI - Identifying effective alternative therapies for common conditions. PMID- 16477886 TI - Medicare Part D: who wins, who loses? AB - The Medicare Part D prescription drug benefit aims to relieve the burden of out of-pocket prescription drug costs for persons older than 65 years, but its effects will vary. Persons with low income and those without prior prescription coverage are projected to save the most, whereas those who lose employer-based coverage are predicted to pay more for their existing regimens. PMID- 16477887 TI - Out-of-pocket prescription costs a continuing burden under Medicare Part D. AB - Of 29 million expected Part D beneficiaries, 6.9 million are projected to have annual out-of-pocket medication expenses greater than dollar 750. Accounting for one fourth of all Part D enrollees, these beneficiaries also are most likely to have high aggregate health care costs, putting them at continued financial risk unless additional policy options are considered. PMID- 16477888 TI - Mind the gap: Medicare Part D's coverage gaps may affect patient adherence. AB - Medicare Part D will lower medication expenditures for many older patients. However, its complex design incorporates a staggered series of cost-sharing mechanisms that create gaps in coverage and may have a negative impact on medication adherence. PMID- 16477889 TI - Are alpha-glucosidase inhibitors effective for control of type 2 diabetes? PMID- 16477890 TI - Gabapentin for pain: balancing benefit and harm. PMID- 16477891 TI - Diagnosis and treatment of community-acquired pneumonia. AB - Patients with community-acquired pneumonia often present with cough, fever, chills, fatigue, dyspnea, rigors, and pleuritic chest pain. When a patient presents with suspected community-acquired pneumonia, the physician should first assess the need for hospitalization using a mortality prediction tool, such as the Pneumonia Severity Index, combined with clinical judgment. Consensus guidelines from several organizations recommend empiric therapy with macrolides, fluoroquinolones, or doxycycline. Patients who are hospitalized should be switched from parenteral antibiotics to oral antibiotics after their symptoms improve, they are afebrile, and they are able to tolerate oral medications. Clinical pathways are important tools to improve care and maximize cost effectiveness in hospitalized patients. PMID- 16477892 TI - Nonhormonal therapies for hot flashes in menopause. AB - Numerous reports in the medical literature and popular media have discussed the effectiveness of various nonhormonal agents in reducing menopausal hot flash symptoms. Data for these therapies are limited, and most of the studies have been conducted in women with a history of breast cancer. Selective serotonin reuptake inhibitors and venlafaxine have been shown to reduce hot flashes by 19 to 60 percent and were well tolerated by study participants. Soy isoflavones reduced hot flashes by 9 to 40 percent in some trials, but most trials showed no difference compared with placebo. Black cohosh and red clover also have had inconsistent results, with some trials showing benefit and some no difference compared with placebo. Soy isoflavones, black cohosh, and red clover were well tolerated in clinical trials. Other agents that have been used to alleviate hot flashes include belladonna/ergotamine tartrate/phenobarbital combination, dong quai, evening primrose oil, gabapentin, ginseng, mirtazapine, trazodone, vitamin E, and wild yam, but few data regarding their effectiveness have been published. Further randomized controlled trials are needed. PMID- 16477893 TI - Information from your family doctor. Nonhormonal options for hot flashes. PMID- 16477894 TI - Evaluation and treatment of constipation in infants and children. AB - Constipation in children usually is functional and the result of stool retention. However, family physicians must be alert for red flags that may indicate the presence of an uncommon but serious organic cause of constipation, such as Hirschsprung's disease (congenital aganglionic megacolon), pseudo-obstruction, spinal cord abnormality, hypothyroidism, diabetes insipidus, cystic fibrosis, gluten enteropathy, or congenital anorectal malformation. Treatment of functional constipation involves disimpaction using oral or rectal medication. Polyethylene glycol is effective and well tolerated, but a number of alternatives are available. After disimpaction, a maintenance program may be required for months to years because relapse of functional constipation is common. Maintenance medications include mineral oil, lactulose, milk of magnesia, polyethylene glycol powder, and sorbitol. Education of the family and, when possible, the child is instrumental in improving functional constipation. Behavioral education improves response to treatment; biofeedback training does not. Because cow's milk may promote constipation in some children, a trial of withholding milk may be considered. Adding fiber to the diet may improve constipation. Despite treatment, only 50 to 70 percent of children with functional constipation demonstrate long term improvement. PMID- 16477895 TI - Information from your family doctor. Chronic constipation in your child. PMID- 16477896 TI - Information from your family doctor. Help for your child's constipation. PMID- 16477897 TI - Pancreatic cancer: diagnosis and management. AB - Although only 32,000 new cases of adenocarcinoma of the pancreas occur in the United States each year, it is the fourth leading cause of cancer deaths in this country. The overall five-year survival rate is 4 percent, and localized, resectable disease has only a 17 percent survival rate. Risk factors include smoking, certain familial cancer syndromes, and familial chronic pancreatitis. The link between risk of pancreatic cancer and other factors (e.g., diabetes, obesity) is less clear. Most patients present with obstructive jaundice caused by compression of the bile duct in the head of the pancreas. Epigastric or back pain, vague abdominal symptoms, and weight loss also are characteristic of pancreatic cancer. More than one half of cases have distant metastasis at diagnosis. Computed tomography is the most useful diagnostic and staging tool. Ultrasonography, magnetic resonance imaging, and endoscopic retrograde cholangiopancreatography may provide additional information. The majority of tumors are not surgically resectable because of metastasis and invasion of the major vessels posterior to the pancreas. Resectable tumors are treated with the Whipple procedure or the pylorus-preserving Whipple procedure. Adjuvant fluorouracil-based chemotherapy may prolong survival. For nonresectable tumors, chemotherapy with gemcitabine prolongs survival. Other agents are being studied. Radiation combined with chemotherapy has slowed progression in locally advanced cancers. Throughout the illness and during end-of-life care, patients need comprehensive symptom control. PMID- 16477898 TI - Screening for peripheral arterial disease: recommendation statement. PMID- 16477899 TI - Vaginal misoprostol for cervical ripening in term pregnancy. PMID- 16477900 TI - Asymptomatic penile rash. PMID- 16477901 TI - At-home genetic tests. PMID- 16477902 TI - Prevention. We've come a long way baby..or have we? PMID- 16477903 TI - Challenges in evidence-based medicine. PMID- 16477904 TI - Updated classification of findings for evaluation of sexual assault in children. PMID- 16477905 TI - When patients begin to trust us. PMID- 16477906 TI - Toxoplasmosis during pregnancy. AB - QUESTION: One of my female patients was tested for Toxoplasma and found positive for immunoglobulin G (IgG). She is planning to get pregnant soon. What are the risks for her and her baby? ANSWER: Up to 25% of Canadians are IgG-positive for Toxoplasma due to past exposure. Immunoglobulin M (IgM) titres indicate current infection. Toxoplasma gondii is an obligate intracellular protozoan that has several forms during its life cycle: oocyte, tachyzoite, and cyst. PMID- 16477907 TI - Dermacase. Pyogenic granuloma. PMID- 16477908 TI - Evidence-based periodic health examination of adults. Memory aid for primary care physicians. AB - PROBLEM ADDRESSED: There is currently no peer-reviewed evidence-based memory aid that incorporates recommended prevention guidelines to direct family physicians during periodic health examination of adults. OBJECTIVE OF PROGRAM: To devise a memory aid to guide primary care physicians during periodic health examination of adults that incorporates the most current evidence-based recommendations of the Canadian Task Force on Preventive Health Care and of the United States Preventive Services Task Force. PROGRAM DESCRIPTION: This memory aid is a two-page easy-to use form that lists evidence-based maneuvers for adults aged 21 to 64 that should be carried out during periodic health examinations. This article describes the form and discusses the evidence currently available for the maneuvers mentioned on the form. To validate the memory aid, results of qualitative assessment in one academic and 15 community settings are presented. CONCLUSION: This user-friendly memory aid was developed to provide primary care physicians with rigorously evaluated guidelines in an accessible format for use during periodic health examination of adults. PMID- 16477911 TI - Family practice-friendly guidelines. Anti-infective guidelines (1994-2005). PMID- 16477909 TI - Preventive care checklist form. Evidence-based tool to improve preventive health care during complete health assessment of adults. AB - PROBLEM ADDRESSED: Preventive care is a cornerstone in the practice of family medicine, but is often difficult to provide because of a lack of time and logistic difficulties. OBJECTIVE OF PROGRAM: To develop an evidence-based practice-relevant preventive care checklist form to be used by family physicians during complete health assessment of adults. PROGRAM DESCRIPTION: Guidelines for preventive health care of adults at average risk from the Canadian Task Force on Preventive Health Care and from other Canadian sources where the Task Force guidelines were not up-to-date were reviewed. Checklist forms covering recommended preventive health care maneuvers were created. The forms incorporate evidence-based preventive care guidelines as well as non-evidence-based components that are a part of routine practice. The forms require few resources to implement, are cost-effective, and are easy to use. The forms list items needed to meet provincial billing requirements for complete health assessments and have space for physicians to make notes. The forms can be used electronically or printed off and photocopied for use in paper-based charts. CONCLUSION: The Preventive Care Checklist Form is a low-cost, easy-to-use tool that merges practice maneuvers with evidence-based recommendations. It could help improve preventive care practices in Canada. PMID- 16477910 TI - [The periodic health examination: a comparison of United States and Canadian recommendations]. AB - OBJECTIVE: To compare recommendations of the Canadian Task Force on Preventive Health Care with those of the United States Preventive Services Task Force for periodic health examinations (PHEs), following the usual sequence of a medical interview. QUALITY OF EVIDENCE: Each task force reviewed the literature exhaustively and created a system of classification that indicated the quality of the evidence. MAIN MESSAGE: Two tables sum up the conclusions of the task forces with respect to preventive measures to be included in PHEs: one for adults generally and one for women specifically. Three other tables show measures for which recommendations are different or conflicting, as well as measures that might be excluded. Several forms and other materials for PHEs based on these comparisons can be found at http://medecinefamiliale.com/umf/emc/emp_guide.htm. Many recommendations are similar; in spite of this, many physicians fail to include them in PHEs. Certain factors could explain the differences between the recommendations, including the challenge of arriving at a standard scientific process for reviewing data, the fact that formulating recommendations is a social as well as a scientific process, and the fact that the CTFPHC is seriously underfunded. CONCLUSION: A scientific review of the literature, even when performed by experts using strict criteria, is not easy to standardize. The differences that our comparison revealed, some of which are substantial, highlight the need to further examine how recommendations are formulated. More research in this field would be helpful. PMID- 16477912 TI - One month in India. An international medical elective. PMID- 16477913 TI - Payment by results: not what it seems. PMID- 16477914 TI - National survey of radiotherapy fractionation practice in 2003. AB - AIMS: To document UK practice in radiotherapy fractionation. METHODS: All radiotherapy centres in the UK participated in a 1-week audit from 29 September 2003. Fractionation data were collected for all patients starting external beam radiotherapy. This included 2498 patients who were prescribed 32 547 fractions. RESULTS: For the radical treatment of non-skin malignancy (n = 708), the prescribed dose ranged from a single fraction of 8 Gy for total-body irradiation to 75 Gy in 43 fractions for prostate cancer. Postoperative treatment for breast cancer was dominated by three regimens: 40 Gy in 15 fractions; 45 Gy in 20 fractions; and 50 Gy in 25 fractions. Palliative treatment was given in a single fraction to 393 patients (36%) with doses of up to 15 Gy. Three hundred and ninety patients (36%) received four to seven fractions delivering 20-25 Gy. Only 89 patients (8%) received more than 10 fractions with palliative intent but used 29% of such fractions. In the treatment of bone metastases, the most common prescriptions were 8-10 Gy in a single fraction and 20 Gy in five fractions. CONCLUSION: UK radiotherapy practice has become more uniform and moved closer to practice in North America and Europe over the past 15 years. For radical radiotherapy, 54% of prescriptions were for a fraction size of 1.8-2.0 Gy but the distribution was bi-modal and 20% of patients were prescribed fraction sizes of 2.7-3.0 Gy. Evidence-based practice now supports hypo-fractionated palliative treatment favouring single fractions for bone metastases and one or two fractions for many patients with advanced lung cancer. Two fractions are advised for some patients with brain metastasis. If these guidelines had been applied uniformly, then the number of treatments prescribed for palliation could have fallen by 36% from 5197 to 3313. This would have represented a 6% reduction in the overall radiotherapy workload. Not all patients are suitable for such hypo-fractionated treatments, but this is an area in which resource use can be improved. In the postoperative management of breast cancer, a change in practice to use 15 fractions uniformly would reduce overall radiotherapy workload by 4%. By contrast, a change to 25 fractions would increase overall workload by 7%. PMID- 16477915 TI - Radiotherapy treatment verification in the UK: an audit of practice in 2004. AB - AIMS: To audit current practice related to treatment verification undertaken in radiotherapy departments throughout the UK. MATERIALS AND METHODS: A questionnaire was circulated to the radiotherapy service managers of 62 radiotherapy centres in the UK. This looked in detail at the department demographics, imaging equipment, site-specific verification protocols, and training and competency assessment of staff responsible for verification. RESULTS: The response rate was 48% (30/62). All departments were using megavoltage imaging equipment in routine clinical practice. Twenty-four out of 29 (83%) departments that had electronic portal imaging capability were using image analysis software for verification. Twenty-nine out of 30 (97%) departments had site-specific written verification protocols. Twenty out of 30 (67%) treatment centres audited set-up errors within their department. Forty-three per cent of centres were using simulator image as the reference image of choice across all sites. Electronic portal imaging, alone or in combination with portal film, was being used for verification in 75% of the centres. Fifty-three per cent of centres used off-line correction strategies for measuring set-up errors across all sites. Radiographer-led interventions were primarily in the pelvis. CONCLUSION: Presently in the UK, verification strategies vary widely at individual treatment sites and between departments. Dedicated departmental verification teams, with input from radiographers, physicists and clinicians, may assist in the effective implementation of evidence-based verification. The inclusion of comprehensive verification protocols within multicentre radiotherapy trials encourages standardisation across treatment centres. PMID- 16477916 TI - Consent for investigating and treating adults with cancer. AB - The importance of the consenting process, as a key activity in patient care, has been recognised by the Department of Health with the production of a policy aimed at ensuring patient focused national standards. Cancer treatments are complex and multi-disciplinary encompassing difficult issues around outcomes and toxicity. This article looks at the process within the UK Cancer network and addresses some of the situations which occur in clinical practice. Examples of difficult scenarios are given to illustrate the application of the basic principles. PMID- 16477917 TI - Evaluation of late toxicities of patients with carcinoma of the cervix treated with radical radiotherapy: an audit from India. AB - AIMS: To evaluate the incidence of, and factors affecting, late toxicities of women with carcinoma of the cervix treated with radical radiotherapy. MATERIALS AND METHODS: Between 1996 and 2001, 1069 women with carcinoma of the cervix (stage I-IVA) were treated at our centre with external-beam radiotherapy (EBRT) and intra-cavitary radiotherapy (ICRT) (n = 871) or EBRT alone (n = 198). Median follow-up was 34 months. Median dose to point A was 81 Gy. RESULTS: Five-year actuarial incidence of overall (all grades) and severe (grade 3/4) late toxicities in the rectum, bladder, small intestine and subcutaneous tissue were 12.3% and 1.1%, 11.2% and 1.2%, 9.2% and 0.2%, and 23.1% and 1.2%, respectively. Vaginal adhesions were seen in 29.6% of cases and stenosis in 33.9% of cases. On multivariate analysis, factors adversely affecting overall incidence of proctitis were anterior-posterior (AP) separation of patient more than 18 cm and presence of comorbid diseases. Presence of comorbid diseases was the only factor affecting the incidence of severe proctitis (grade 3/4). AP separation more than 18 cm adversely affected the incidence of cystitis, both overall and severe. Late toxicities (all grades) in small bowel were increased in subsets, like women younger than 50 years and women with comorbid diseases, but no factor emerged as significant for incidence of severe toxicities. Subcutaneous fibrosis was significantly higher in patients with AP separation over 18 cm, those treated by cobalt machines and those who received EBRT only. Severe subcutaneous fibrosis was influenced by the use of EBRT alone. Overall incidence of vaginal toxicity was higher in women whose overall treatment time (OTT) was shorter and in women who received ICRT. Vaginal stenosis was higher in elderly women and in women who received ICRT by low dose rate. CONCLUSIONS: Even with telecobalt machines, impressive results with acceptable late toxicity can be achieved in the treatment of cancer of the cervix using an ideal combination of EBRT with ICRT. PMID- 16477918 TI - Improved survival after concurrent weekly cisplatin and radiotherapy for cervical carcinoma with assessment of acute and late side-effects. AB - AIMS: A recent meta-analysis has shown a survival advantage for the addition of concurrent chemotherapy to radiotherapy in the treatment of cervical carcinoma. Controversy persists about the most appropriate chemotherapy schedule and whether similar results for tumour control and toxicity may be achieved with optimally delivered radiotherapy. A single-centre experience of a concurrent chemotherapy regimen is presented. MATERIALS AND METHODS: All women treated with concurrent chemoradiotherapy at a university hospital from 1 January 1999 to 1 May 2002 were identified. Acute and late complications were scored using the National Cancer Institute Common Toxicity Criteria and RTOG/ EORTC system, respectively. Univariate and multivariate analyses were carried out to examine the relationship between demographics, stage, overall treatment time, radiotherapy dose, selectron insertion, number of chemotherapy cycles and occurrence of acute and late toxicity. RESULTS: Seventy-nine women received concurrent weekly cisplatin (40 mg/m2) with radiotherapy. Thirty-eight per cent had early tumours (FIGO IIA or less) and 62% had locally advanced tumours. Twenty-eight per cent of women had surgery as part of primary treatment. Radiation technique included external-beam pelvic radiotherapy (EBRT) (45-50.4 Gy in 25-28 fractions) and medium-dose rate brachytherapy single insertion (25-27 Gy to point A) or EBRT alone. Median overall treatment time was 49 days (range 23-91 days). Three-year survival rate was 87% (95% confidence interval [CI] 79-95%). Three-year, progression-free survival rate was 75% (95% CI 65-85%). At a median follow-up of 35 months: 27 (34%) women experienced 45 episodes of acute grade 3 or 4, and eight women (10%) experienced 12 late grade 3 or 4 complications. CONCLUSIONS: Weekly cisplatin 40 mg/m2 concurrent with radiotherapy is well tolerated when given to an unselected population of patients. Survival rates seem to be excellent, with both local control and overall survival being at least as good as those in published randomised trials. PMID- 16477919 TI - Results of external-beam radiotherapy alone in invasive cancer of the uterine cervix: a retrospective analysis. AB - AIMS: In this retrospective audit, we describe the results of external-beam radiotherapy (EBRT) alone in patients with invasive cancer of the cervix treated at our centre. MATERIAL AND METHODS: We included 146 patients with invasive cancer of the cervix who were treated with EBRT to a total dose of 60-66 Gy between January 1996 and December 2001. None of these patients were suitable for intracavitary radiotherapy (ICRT) after a median dose of 46 Gy. A boost dose of 14-20 Gy was given after a gap of 2-4 weeks. Most patients belonged to stage IIIB (n = 124). RESULTS: Follow-up of patients at risk ranged from 19 to 89 months (median 48 months). One hundred and thirty-six patients (93.2%) received EBRT to a dose of 66 Gy, and 10 patients (6.8%) received 60 Gy. Overall treatment time (OTT) ranged from 56 to 160 days (median 78 days). At completion of 46 Gy of EBRT, 63 patients achieved partial response and 83 patients had stable disease. Five-year overall survival, disease-free survival (DFS) and pelvic control were 15.1% (median 9 months), 11.6% (median 5 months) and 21.9% (median 6 months), respectively. Factors found to affect 5-year pelvic control in univariate analysis by Kaplan-Meier method were response to EBRT at 46 Gy (partial response 36.5% and stable disease 10.8%), age (> or = 50 years 28.8% and < 50 years 13.6%) and OTT (< 90 days 26.5% and > or = 90 days 12.5%). For DFS and overall survival, response to EBRT was the only factor that was significant in univariate analysis. In multivariate analysis by Cox's proportional hazard model, response to EBRT was the only factor to influence pelvic control (P = 0.007), DFS (P = 0.01) and overall survival (P < 0.001). CONCLUSIONS: Overall outcome of patients in whom ICRT was not given remains less than satisfactory. Response to EBRT emerged as the most important factor to predict all clinical outcomes. To improve upon the dismal results of EBRT alone, we will have to decrease the OTT and consider concurrent chemo-radiation with cisplatin. PMID- 16477920 TI - Radical radiotherapy for bladder cancer: retrospective analysis of a series of 459 patients treated in an Italian institution. AB - AIMS: To contribute to the available evidence about the efficacy of exclusive radiotherapy for bladder cancer through a retrospective analysis of a large series of patients consecutively treated in a single institution. MATERIALS AND METHODS: A total of 459 patients with UICC categories T1-T4, N0-Nx and M0 bladder cancer consecutively treated with radiotherapy alone with radical intent formed the clinical basis for this study. Many of them (and particularly the T1 cases) had poor medical conditions or were unfit for surgery. About half of the cases (54%) had a T2 tumour, and about 18% had T3-T4 disease. Eighty per cent of the cases received minimal doses in the target volume in the range 60-70 Gy; pelvic lymph nodes were treated in 34%. Simple radiotherapy techniques were used in most cases. Average follow-up for living patients was 4.4 years. Results were analysed according to number and type of relapses: overall survival, disease-specific survival, failure-free survival probability, acute and late toxicity (RTOG scale). RESULTS: Actuarial 5-year overall survival, disease-specific survival and failure-free survival rates at 5 years for the entire series were 36%, 56%, 33%, respectively. Age, T category (for all the end points) and tumour dose (only for failure-free survival) were significantly related to prognosis at multivariate survival analysis. Late enteric toxicity (6.1% of the cases) was significantly linked with the treated volumes (univariate analysis). Urinary late toxicity (23% of cases) was linked with age and T category (multivariate analysis). In both cases, toxicity was mostly Grade 1 or 2. CONCLUSIONS: The results of radiotherapy in this negatively selected series, accrued over a long period of time in patients treated with unsophisticated techniques, are reasonably good; they add to the evidence available to support the use of modern bladder-sparing programmes, including the association of chemo- and radiotherapy. PMID- 16477921 TI - Perioperative allogeneic blood transfusion, the related cytokine response and long-term survival after potentially curative resection of colorectal cancer. AB - AIMS: It is still debated whether perioperative blood transfusion alters the incidence of disease recurrence or otherwise affects the prognosis after curative resection of malignant tumours. We conducted a prospective observational study of patients with colorectal cancer to provide data on the effect of blood transfusion and the related perioperative cytokine response on long-term prognosis. MATERIALS AND METHODS: Perioperative blood samples were obtained from 117 patients with colorectal cancer undergoing potentially curative resection. Factors associated with perioperative blood transfusion were assessed, and their relationship with early postoperative systemic responses of tumour growth factors and long-term prognosis were evaluated. RESULTS: Independent factors associated with perioperative blood transfusion were preoperative anaemia, operative blood loss and the development of postoperative infectious complication. The patients receiving transfusions were subdivided according to the independent factors. Group A comprised 19 patients who received blood transfusions because of preoperative anaemia and Group B comprised 16 patients who received blood transfusions because of excessive operative blood loss. Group B patients showed exaggerated postoperative systemic induction of interleukin (IL)-6 and IL-6 triggered tumour growth factors, such as hepatocyte growth factor and vascular cell adhesion molecule-1. Intraoperative blood transfusion under intense surgical stress was associated with poor prognosis, whereas preoperative blood transfusion for correcting anaemia or intraoperative blood transfusion under less invasive surgery was not associated with survival. Multivariate analysis using the Cox proportional hazards method showed that a significant independent risk was demonstrated for blood transfusion, T stage, lymph-node metastasis and perioperative peak levels of IL-6. CONCLUSION: Blood transfusion and intense surgical stress might synergistically affect the long-term prognosis after curative resection of colorectal cancer. Postoperative exaggerated systemic inductions of IL-6 may indicate the critical situation that could lead to disease recurrence. PMID- 16477922 TI - Quality of life and symptom end points in palliative bone metastases trials. PMID- 16477923 TI - A pilot randomised comparison of dexamethasone 96 mg vs 16 mg per day for malignant spinal-cord compression treated by radiotherapy: TROG 01.05 Superdex study. AB - AIM: To test the viability of a full-scale randomised comparison of two steroid doses given with radiotherapy for malignant spinal-cord compression (MSCC), to test Internet randomisation and to compare different functional outcome measures. MATERIALS AND METHODS: A log of screened patients at eight recruiting centres was maintained. Patients were randomised via the Superdex website to either 96 mg or 16 mg daily of dexamethasone. Radiotherapy treatment was 30 Gy in 10 fractions. Outcomes assessed used ambulation, Barthel Index ambulation, Functional Independence Measure (FIM) ambulation and Functional Improvement Score (FIS) at 1 month. RESULTS: One hundred and thirty-one patients were screened. Ninety-three (71%) were ineligible, 65% of these were because duration of prior steroid use was greater than 12 h, failure to meet strict definition of magnetic resonance imaging, defined MSCC, multi-level disease or previous spinal-cord compression treatment. Twenty of the 38 eligible patients were randomised, including seven outside standard office hours. There was a high rate of serious adverse events (n = 9), but only one was considered likely to be related to study medication. At baseline, 75% were ambulant, 70% had FIM ambulation scores greater than 5 and 50% had Barthel Index ambulation scores greater than 2. At day 28, including all randomised patients (by scoring four dead patients as non-ambulant), ambulation scores by the various definitions were 60%, 45% and 40%, respectively. For the 16 patients evaluable at day 28, the mean FIS was -1.4. Median survival was 69 days and 1-year survival 13%. CONCLUSION: Web randomisation was successful; however, the high ineligibility rate precludes a full-scale dexamethasone dose trial in Australia. Choice of measure of ambulation has potentially significant effects on outcomes and implications for the design of any future MSCC trials. Referral delays are of concern. PMID- 16477924 TI - Carboplatin and hypofractionated accelerated radiotherapy: a dose escalation study of an outpatient chemoradiation schedule for squamous cell carcinoma of the head and neck. AB - AIM: To examine the feasibility and determine the maximum tolerated dose of outpatient carboplatin given with synchronous hypofractionated accelerated radiotherapy for squamous cell carcinoma of the head and neck (SCCHN). MATERIALS AND METHODS: Patients with stages II-IV SCCHN and unresected primary tumour were treated with synchronous carboplatin given in an outpatient setting on day 1 and day 21 in cohorts of three to six patients with incremental area under curve (AUC) factors commencing at 3.5. Grade 3 mucositis persisting for 4 weeks in two patients in a cohort was considered dose limiting. RESULTS: A total of 19 patients were enrolled and assessable for toxicity. All 19 patients completed 55 Gy of radiotherapy and were assessable for response. Grade 3 mucositis lasting 4 weeks or more was seen in three patients, two of them received AUC 5 carboplatin. A complete response was seen in 16 patients, with a further patient having a partial response, giving a response rate of 89%. With a median follow-up of 24 months (range 11-30 months), 13 patients were alive with no evidence of recurrent disease. Local recurrence had occurred in four patients with distant spread in three patients. CONCLUSION: Carboplatin with concurrent hypofractionated accelerated radiotherapy is feasible for patients with advanced SCCHN and good performance status. The recommended phase II dose of carboplatin given in week 1 and week 4 with 55 Gy in 20 fractions is AUC 4.5. PMID- 16477925 TI - Radical radiation therapy in a patient with head and neck cancer and severe Parkinson's disease. AB - We report the case of a 61-year-old man with severe Parkinson's disease, who was successfully treated with postoperative radiation therapy for a T4N2cM0 transglottic squamous cell carcinoma of the larynx, with the aid of a deep brain stimulator. PMID- 16477926 TI - Gemcitabine-induced radiation necrosis of the pectoralis major muscle. PMID- 16477927 TI - The volume of brain metastases may be of prognostic significance in patients with non-small-cell lung cancer classified as RTOG-RPA classes 2 and 3. PMID- 16477928 TI - Chemotherapy and radiotherapy causing radiation myelopathy: what is to blame? PMID- 16477929 TI - Response to thalidomide in chemotherapy-resistant cutaneous T-cell lymphoma. PMID- 16477930 TI - Multiple positron-emission tomography false positives in a patient with malignant melanoma. PMID- 16477931 TI - Capecitabine plus rofecoxib show no activity in patients with metastatic neuroendocrine tumours. PMID- 16477932 TI - Subdural haematoma secondary to carcinoma of the uterine cervix. PMID- 16477933 TI - Toxic epidermal necrolysis after PCV combination chemotherapy for relapsed B-cell lymphoma. PMID- 16477934 TI - Gemcitabine-induced chronic systemic capillary leak syndrome: a life-threatening disease. PMID- 16477935 TI - Uncertainty of measurement: approaches and open problems. AB - Despite of exhaustive literature information, analysts are still fronting serious problems in estimating the uncertainty of measurement (UOM), mainly because of the intrinsic difficulties in choosing an appropriate approach and/or of an inadequate background in statistics and metrology fundamentals. Sometimes, additional problems, at present under debate, make their efforts even more stressful. In this paper, attention is mainly paid to intra-laboratory estimation of UOM, the most frequent situation. After a short re-examination of the existing UOM approaches, some applicative problems are discussed in the light of the most representative literature information. PMID- 16477936 TI - Partitioning of metals between the aqueous phase and suspended insoluble material in fog droplets. AB - This paper discusses the partitioning of metals (K, Na, Ca, Mg, Al, Cu, Fe, Pb and Zn) between the aqueous phase and the suspended insoluble material in fog samples collected in the Po Valley during two extensive fields campaigns. Metals represent on average 11% of the mass of suspended insoluble matter, while the main component is carbon (both organic carbon, OC = 35%, and black carbon, BC = 8%). The unaccounted suspended matter mass is very high, on average 46%, and is attributable to non metallic species, such as O and N and of Si. The principal metals in the insoluble suspended fraction are Fe and Al (2-5%), while the contributions of other metals (Na, Mg, Cu, Pb and Zn) are lower than 1%. Ca and K exhibited high blank values and could not be detected above blank detection limit threshold. The main components in the aqueous phase are NO3- (34%), WSOC (23%), SO4(2-) (18%) and NH4+ (19%), while trace metals and remaining cations and anions accounted for less than 1% of solute mass. The main dissolved trace metals in fog droplets are Zn, Al and Fe, while the main metallic cations are Na and Ca. Fe and Al are the only metals preferentially distributed in the suspended insoluble matter of fog droplets (partitioning ratio respectively 37% and 33%). All other metals are mostly dissolved in the aqueous phase (mean partitioning ratios of Mg, Pb, Zn, Cu and Na are 69%, 70%, 77%, 81% and 87%). These findings are in agreement with literature data on metal speciation in cloud and rain samples. The dependence of partitioning ratios on pH is investigated for the different metals, with only Al showing a clear partitioning ratio decrease with increasing pH. Conversely, the other metals show no dependence or a complex and highly variable behaviour. The partitioning ratio of iron (mean 37%) observed in the Po Valley fog samples is much higher than the water extractable iron in aerosol particles (typically 1-2 %): this fact can be explained by differences in the aerosol sources and composition among sites and by chemical processes in the aqueous phase, such as complexation and redox reactions involving organic ligands (oxalate, or other organic acids as humic-like organic matter) which may promote Fe solubility. PMID- 16477937 TI - Prospects of second generation artificial intelligence tools in calibration of chemical sensors. AB - Multivariate data driven calibration models with neural networks (NNs) are developed for binary (Cu++ and Ca++) and quaternary (K+, Ca++, NO3- and Cl-) ion selective electrode (ISE) data. The response profiles of ISEs with concentrations are non-linear and sub-Nernstian. This task represents function approximation of multi-variate, multi-response, correlated, non-linear data with unknown noise structure i.e. multi-component calibration/prediction in chemometric parlance. Radial distribution function (RBF) and Fuzzy-ARTMAP-NN models implemented in the software packages, TRAJAN and Professional II, are employed for the calibration. The optimum NN models reported are based on residuals in concentration space. Being a data driven information technology, NN does not require a model, prior- or posterior- distribution of data or noise structure. Missing information, spikes or newer trends in different concentration ranges can be modeled through novelty detection. Two simulated data sets generated from mathematical functions are modeled as a function of number of data points and network parameters like number of neurons and nearest neighbors. The success of RBF and Fuzzy-ARTMAP-NNs to develop adequate calibration models for experimental data and function approximation models for more complex simulated data sets ensures AI2 (artificial intelligence, 2nd generation) as a promising technology in quantitation. PMID- 16477938 TI - Fluoride wastewater treatment by adsorption onto metallurgical grade alumina. AB - The adsorption of fluoride onto metallurgical grade alumina (to produce Al) was investigated under different conditions: pH, contact time and adsorbent concentration. Data were evaluated with the aim of developing an alternative treatment technology for washing wastewater arising from an Aluminum can production plant. Kinetics and adsorption isotherms data have been also produced Sorption is greatly affected by pH and the best condition for fluoride removal are obtained at pH 5-6 and alkalinity competes successfully with fluoride ions for the exchange sites. Experiments with fixed beds indicate that fluoride is removed from wastewater by metallurgical-grade alumina with a capacity of 12.21 mg of F per gram and adsorption increases of about 25% at appropriate pH. The Mass Transfer Zone at 5% of the breakthrough occupies 70.6% of the total column length. The optimization of aluminum precipitation by pH adjustment and with different precipitant agents has been done. PMID- 16477939 TI - On the hydrolysis of the dioxouranium(VI) ion in oxalate solutions. AB - The complex formation between the dioxouranium (VI) and the oxalate ions has been investigated by measuring the potential of a glass electrode, at 25.00 degrees C, in 1 and 3 M NaClO4, at lower acidities than 10(-4.5) M, in order to favour the formation of (mixed) ternary species. The upper limits of concentration of all the analytical parameters have been imposed by the modest solubility of Na2C2O4 in the ionic media. The experimental measurements at different ionic strengths have been treated by means of the computerised least square programme LETAGROP - ETITR. Ternary complexes of general composition (p, q, r) are formed according to reaction (1), in addition to the already reported binary complexes. pUO2(2+) + qH2O + rC2O4(2-) <==> (UO2)p(OH)q(C2O4)r(2p-q-2r)(+) + qH+ (1). The stoichiometric compositions of the ternary species are (1, 1, 1), (2, 4, 2), (2, 2, 4). Their formation constants, expressed in molality, obtained in the two ionic media, are listed below. [table: see text]. For reasons discussed in the present work in the last column only the value of the constant in 1 M ionic medium is reported for the species (2, 4, 2). Additional evidence on the stoichiometric composition of the species formed is afforded by the mass spectrometric measurements, collected in solutions of known composition. PMID- 16477940 TI - Slurry sampling techniques for the determination of lead in Bangladeshi fish samples by electrothermal atomic absorption spectrometry with a metal tube atomizer. AB - Ultrasonic slurry sampling electrothermal atomic absorption spectrometry with a metal tube atomizer has been applied to the determination of lead in Bangladeshi fish samples. The slurry sampling conditions, such as slurry stabilizing agent, slurry concentration, pyrolysis temperature for the slurried fish samples, particle size and ultrasonic agitation time, were optimized for electrothermal atomic absorption spectrometry with the Mo tube atomizer. Thiourea was used as the chemical modifier for the interference of matrix elements. The detection limit was 53 fg (3S/N). The determined amount of lead in Bangladeshi fish samples was consistent with those measured in the dissolved acid-digested samples. The advantages of the proposed methods are easy calibration, simplicity, low cost and rapid analysis. PMID- 16477941 TI - Photocatalytic degradation of high ammonia concentration water solutions by TiO2. AB - Recent studies reported the use of suspended TiO2 powder as photo catalyst in the degradation process of several organic and inorganic substances in water, like chlorinated hydrocarbons, alkyl- and alkanoloammines and ammonia. In order to evaluate the possibility of using this method for a further removal of ammonia from wastewater after a biological nitrification/denitrification treatment, a kinetic study of photo oxidation over TiO2 of NH3/NH4 in the high concentration range of 26 - 214 mg/I, has been carried out. Effects of pH, TiO2, ammonium and dissolved O2 concentration on the degradation efficiency and reaction products distribution (NO2-, NO3- e N2) were studied. The results have shown that, at a catalyst concentration of 0.012% (p/v), the photo catalytic oxidation of ammonia can be described with the Langumuir- Hinshelwood kinetic model, as reported by several authors. PMID- 16477942 TI - Validated spectrophotometric and fluorimetric methods for analysis of clozapine in tablets and urine. AB - Five spectrophotometric methods and one fluorimetric method have been developed and validated for the analysis of clozapine. The spectrophotometric methods were based on the charge-transfer complexation reaction between clozapine as electron donor and each of iodine as sigma-acceptor or 7,7,8,8-tetracyanoquinondimethane (TCNQ), 2,3-dichloro-5,6-dicyano-1,4-benzo-quinone (DDQ), tetracyanoethane (TCNE), and p-chloranilic acid (pCA) as pi-acceptors. The obtained complexes were measured spectrophotometrically at 365, 843, 460, 414, and 520 nm for iodine, TCNQ, DDQ, TCNE, and pCA, respectively. The fluorimetric method was based on the oxidation of clozapine in the presence of perchloric acid by cerium (IV), and subsequent measuring the fluorescence of the produced cerium (III) fluorimetrically at lambda(excitation) 260 and lambda(emission) 355 nm. Under the optimum assay conditions, Beer's law was obeyed at concentrations ranged from 4 200 microg mL(-1) for the spectrophotometric methods and from 24-250 ng mL(-1) for the fluorimetric method. The limits of detection for the spectrophotometric methods were 1.12, 1.76, 2.22, 0.95, and 13.26 microg mL(-1) for iodine, TCNQ, DDQ, TCNE, and pCA, respectively. The limit of detection for the fluorimetric method was 6.69 ng mL(-1). The proposed methods were successfully applied to the analysis of clozapine in tablets with good recoveries. The fluorimetric method could also be applied to the analysis of clozapine in spiked urine samples. The molar ratios and the reaction mechanisms were investigated. PMID- 16477943 TI - Evaluation of groundwater quality within a typical Egyptian village, North of Cairo, Egypt. AB - Groundwater is the second most abundant water resource in Egypt, 2/3 of which is consumed within the Nile Delta Region and the valley. However, with recent intensification of agricultural and urbanizing activities over the past 40 years, the water quality in the Nile basin aquifer has significantly deteriorated. The objective of this study is to survey the variation in the groundwater quality tapped within a typical Egyptian village located in the valley, north of Cairo, from January until April 2000. Results revealed that locations close to agricultural lands reflected an increase in the TDS, ammonia, nitrates, chlorides and sulfates contents as a implication of vertical and side infiltration of multiple pollutants. In addition, locations within residential quarters recorded significant BOD and COD concentrations, reflecting a significant organic matter intrusion at these wells. Multivariate analysis for the January data indicated that agricultural activities had a greater impact on these waters causing an overall increase in TDS and EC contents. However, with the increase in Nile surface water flow during April, there was an increase in fresh water infiltration within this area, which helped to reduce the TDS, EC and other inherent ionic constituents of these waters. PMID- 16477944 TI - Determination of diphenamide, napropamide and metolachlor in tobacco by gel permeation chromatographic clean-up and high performance liquid chromatography. AB - Diphenamide, napropamide and metolachlor (FIG. 1) are selective, pre-emergence arylamide herbicides used to control the growth of annual grasses and broadleaf weeds in a variety of fields, e.g. fruit trees, nuts, corns, green crops, etc. They possess high activity and moderate toxicity. For food and environment safety, the detailed investigations on their residues and metabolism are very important. Diphenamide, napropamide and metolachlor in the pesticide products, serum, urine, soil, environmental water, fruits and wine have been widely analyzed by ELISA, fluorescence, phosphorescence, capillary electrophoresis, high performance liquid chromatography (HPLC), gas chromatography(GC) and GC mass spectrometry (GC-MS). However, to our knowledge, simultaneous residue analysis of diphenamide, napropamide and metolachlor in tobacco samples has not been extensively documented. Tobacco is greatly consumed by smokers throughout the world. The pesticide residue in tobaccos might be potentially harmful to smokers' health. With this in mind the residue determination and control of diphenamide, napropamide and metolachlor in the tobacco leaves are very important for tobacco products and consumers. For these three herbicides, the tolerable maximum residue limits (MRLs) have been limited ranging from 0.05 (for tobacco products) to 5 mg/kg (for tobacco leaves) in different European countries. For the complex tobacco samples, the GC and HPLC with UV detection suffer from matrix interference making quantification and identification of these herbicides difficult. In such cases the removal of the matrix effects and identification of the target compounds are of great importance. The present work reports the extraction and clean up procedures, as well as, the chromatographic conditions developed for the simultaneous determination of diphenamide, napropamide and metolachlor residues in the fluecured tobacco leaves, from the different sources using HPLC-UV method. PMID- 16477946 TI - Modelling risks in disease mapping. AB - In this article, we propose a strategy of analysis of mortality data with the aim of providing a guideline for epidemiologists and public health researchers to choose a reasonable model for estimating mortality (or incidence) risks. Maps displaying the crude mortality rates or ratios are usually misleading because of the instability of the estimators in low populated areas. As an alternative, many smoothing methods have been presented in the literature based on Poisson inference. They account for the extra-Poisson variation (overdispersion), frequently present in the homogeneous Poisson model, by incorporating random effects. Here, we recommend to test for the potential sources of extra-Poisson variation because, depending on them, the models which fit better the data may be different. Overdispersion can be mainly due to spatial autocorrelation, unstructured heterogeneity or to a combination of these two, and also, when studying very rare diseases, it can be due to an excess of zeros in the data. In this article, different situations the analyst may encounter are detailed and appropriate procedures for each case are presented. The alternative models are illustrated using mortality data provided by the Statistical Institute of Navarra, Spain. PMID- 16477945 TI - Comparison of various statistical methods for identifying differential gene expression in replicated microarray data. AB - DNA microarray is a new tool in biotechnology, which allows the simultaneous monitoring of thousands of gene expression in cells. The goal of differential gene expression analysis is to identify those genes whose expression levels change significantly by the experimental conditions. Although various statistical methods have been suggested to confirm differential gene expression, only a few studies compared the performance of the statistical tests. In our study, we extensively compared three types of parametric methods such as T-test, B statistic and Bayes T-test and three types of non-parametric methods such as samroc, significance analysis of microarray and a modified mixture model using both the simulated datasets and the three real microarray experiments. PMID- 16477947 TI - Weighted confidence interval construction for binomial parameters. AB - Confidence intervals, in general, have become an important aspect of reporting statistical results. In particular, interval estimators for binomial proportions have been studied extensively in recent literature. The large-sample Wald intervals are known to perform poorly, but the Wilson intervals have been shown to perform well in a variety of situations. One criticism is the relative difficulty of computing the Wilson or quadratic intervals in comparison to the Wald intervals. We offer a computational formula for the Wilson intervals that is a weighted estimator of the observed proportion, p, and that based on an uninformative prior, 1/2. This contribution enhances our understanding of the coverage behavior of the Wilson intervals. In addition, we contrast the Wilson intervals with other well-known intervals for the case of zero successes. PMID- 16477948 TI - Multi-level zero-inflated poisson regression modelling of correlated count data with excess zeros. AB - Count data with excess zeros relative to a Poisson distribution are common in many biomedical applications. A popular approach to the analysis of such data is to use a zero-inflated Poisson (ZIP) regression model. Often, because of the hierarchical study design or the data collection procedure, zero-inflation and lack of independence may occur simultaneously, which render the standard ZIP model inadequate. To account for the preponderance of zero counts and the inherent correlation of observations, a class of multi-level ZIP regression model with random effects is presented. Model fitting is facilitated using an expectation-maximization algorithm, whereas variance components are estimated via residual maximum likelihood estimating equations. A score test for zero-inflation is also presented. The multi-level ZIP model is then generalized to cope with a more complex correlation structure. Application to the analysis of correlated count data from a longitudinal infant feeding study illustrates the usefulness of the approach. PMID- 16477949 TI - Testing non-additivity (interaction) in two-way ANOVA tables with no replication. AB - Testing for any significant interaction between two variables depends on the number of replicates in each cell of the two-way table and structure of the interaction. If there is interaction between two factors model of observations include interaction term and is called 'non-additive model' which makes interaction and non-additivity equivalent in terms of meaning. When there are several observations taken at each level combination of two variables, testing non-additivity can easily be done by usual two-way ANOVA method which cannot be used when there is only one observation per cell. For the cases with only one observation per cell, some methods have been developed starting with Tukey's one degree-of-freedom test in which interaction is supposed to be the product of two factor's effects. There are other methods which are used for different structures of interaction when there is only one observation. In this paper, we review some of these tests. After presenting general methodology for the two-factor linear model with interaction effect and the general two-way ANOVA method when there are n > 1 observations per cell, we present some methods for testing non-additivity when there is only one observation per cell. Finally, we illustrate these methods on examples. PMID- 16477950 TI - Stimulants and narcolepsy. PMID- 16477951 TI - The case for trait determinants of arousal and sleepiness. PMID- 16477952 TI - Risks of high-dose stimulants in the treatment of disorders of excessive somnolence: a case-control study. AB - STUDY OBJECTIVES: To ascertain complications associated with high-dose stimulant therapy in patients with narcolepsy or idiopathic hypersomnia. DESIGN: Case control, retrospective chart review. SETTING: Sleep center in an academic hospital. PATIENTS: 116 patients with narcolepsy or idiopathic hypersomnia were individually matched by sex, diagnosis, age of onset, and duration of follow-up from both onset and diagnosis. Members of the high-dose group (n = 58) had received at least 1 stimulant at a dosage > or = 120% of the maximum recommended by the American Academy of Sleep Medicine Standards of Practice Committee. The standard-dose control group (n = 58) had received stimulants at a dosage < or = 100% of the American Academy of Sleep Medicine guidelines. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: The prevalence of psychosis (odds ratio = 12.0 [1.6 92.0]), alcohol or polysubstance misuse (odds ratio = 4.3 [1.2-15.2]), and psychiatric hospitalization (odds ratio = 3.2 [1.1-10.0]) was significantly increased in the high-dose group. More high-dose patients also experienced tachyarrhythmias (odds ratio = 3.3 [0.92-12.1] and anorexia or weight loss (odds ratio = 11.0 [1.4-85.2]). The frequency of physician-diagnosed depression, drug seeking and suicide-related behaviors, hypertension, and cardiovascular disease did not differ significantly between the groups. CONCLUSIONS: This study demonstrated a significantly higher occurrence of psychosis, substance misuse, and psychiatric hospitalizations in patients using high-dose stimulants compared to those using standard doses. Tachyarrhythmias and anorexia or weight loss were also more common in this group as compared with controls. Clinicians should be very cautious in prescribing dosages that exceed maximum guidelines. PMID- 16477953 TI - Dose-response effects of tiagabine on the sleep of older adults. AB - STUDY OBJECTIVES: To evaluate the dose- response effects of tiagabine on sleep and safety measures in healthy older adults. DESIGN: Randomized, double-blind, Latin-square design. SETTING: Sleep laboratory. PARTICIPANTS: Twenty-four healthy older adults (11 men, 13 women; mean age 68.0 +/- 6.2 years) INTERVENTIONS: Tiagabine 2, 4, or 8 mg, or placebo, each given on two consecutive nights. MEASUREMENTS AND RESULTS: Polysomnography revealed that compared with placebo, tiagabine 4 mg increased total sleep time, reduced wake after sleep onset, and increased minutes of slow-wave sleep. Tiagabine 8 mg decreased wake after sleep onset, increased slow-wave sleep, and improved a sleep-continuity index. No differences were seen between the 2-mg dose and placebo. Subjective ratings indicated fewer awakenings with the 8-mg dose. Central nervous system adverse events were somewhat higher in the 8-mg condition only. Measures of morning performance were minimally affected. CONCLUSIONS: Research with tiagabine at dosages of 8 mg or less appears warranted in elderly clinical populations. PMID- 16477954 TI - Acute intravenous administration of morphine perturbs sleep architecture in healthy pain-free young adults: a preliminary study. AB - STUDY OBJECTIVES: Pain is a leading cause of sleep disturbances in medical illness. Providing effective analgesia is considered an important intervention to reduce these sleep disturbances. Opioids remain the treatment of choice to relieve postoperative pain in hospitalized patients. However, their effects on sleep in pain patients or normal subjects remain unclear, as previous studies have been conducted mainly with former opioid addicts. The purpose of this investigation was to evaluate and describe the effects of acute clinical doses of morphine on sleep in healthy pain-free subjects. DESIGN: Subjects were randomly assigned to untreated (baseline), morphine (intravenous injections of 0.1 [DOSAGE ERROR CORRECTED] mg/kg), and placebo (intravenous injections of 0.9% NaCI) conditions. SETTING: Sleep laboratory. PARTICIPANTS: Seven healthy pain-free, nonaddicts (5 women, 2 men; mean age = 25 +/- 1.6 years). MEASUREMENTS AND RESULTS: Standard polysomnographic sleep and respiratory variables were measured during 3 experimental conditions. The treatment effect was analyzed with a Latin square cross-over design followed, when appropriate, by Tukey contrasts. Morphine altered sleep architecture by reducing slow-wave sleep (non-rapid eye movement stages 3-4) and rapid eye movement sleep, and by increasing non-rapid eye movement stage 2 sleep. Results did not reveal any statistical differences for other sleep and respiratory variables. CONCLUSIONS: Similar to earlier findings in animals, nondependent opiate addicts, and postoperative patients, morphine was found to reduce duration of slow-wave sleep. Unlike previous reports, however, its acute administration produced a moderate reduction in rapid eye movement sleep and did not increase correlates of arousal (ie, awakenings, electroencephalogram arousals, wake after sleep onset). Future studies should correlate these findings in patients with pain and evaluate whether optimal pain relief with opioid therapy can improve sleep disturbances in pain patients. PMID- 16477955 TI - Performance and cardiovascular measures in normal adults with extreme MSLT scores and subjective sleepiness levels. AB - STUDY OBJECTIVES: The purpose of this study was to determine the relationship of subjective and objective sleepiness across several nights. Extreme groups were chosen based upon both Multiple Sleep Latency Test (MSLT) findings and report of characteristic subjective sleepiness, and groups were compared across sleep, demographic, performance, and physiologic variables. DESIGN AND SETTING: Subjects spent 3 baseline nights and the following days in the laboratory. Standard polysomnographic recordings were made on each night. On each day, subjects had an MSLT, performance testing, and metabolic and heart rate observation periods. PARTICIPANTS: Participants were 50 adult normal sleepers. INTERVENTIONS: None. MEASUREMENT AND RESULTS: Those subjects with sleep latencies on the MSLT of more than 10 minutes following the adaptation night (Alert) were compared with 2 groups of subjects with sleep latencies on the MSLT of less than 7 minutes following the adaptation night. Subjects with MSLT < 7 were divided into those who reported subjective sleepiness during the day (subjective sleepiness > 1 SD above the mean for the entire group-Sleepy-Sleepy) and those who did not report subjective sleepiness (subjective sleepiness < 1 SD above the mean for the entire group--Sleepy-Alert). The Alert group maintained longer sleep latencies than the other groups and had improved performance on vigilance compared to the Sleepy Sleepy group on all days and on some days compared to the Sleepy-Alert group. Vigilance was improved in the Sleepy-Alert group compared with the Sleepy-Sleepy group on all days. The Alert group had higher heart rate and increased low/high spectral heart rate power compared to both sleepy groups, and the Sleepy-Alert group had higher heart rate and increased low/high spectral heart rate power compared to the Sleepy-Sleepy group at some points. CONCLUSIONS: It was concluded that normal adults with short MSLT latencies differ from those with longer latencies on both cardiac and performance variables. Also, those individuals with short latencies can be divided into subgroups claiming subjective sleepiness or denying sleepiness. Those denying sleepiness have improved vigilance performance and greater heart rate and low/high spectral heart rate power compared to those with subjective sleepiness. Both the MSLT group differences and the subjective group differences imply that ability to maintain wakefulness and performance in sedentary situations may be related to innate ability to maintain physiologic arousal. PMID- 16477956 TI - Cold exposure and sleep in the rat: effects on sleep architecture and the electroencephalogram. AB - STUDY OBJECTIVES: Acute exposure to low ambient temperature modifies the wake sleep cycle due to stage-dependent changes in the capacity to regulate body temperature. This study was carried out to make a systematic analysis of sleep parameters during the exposure to different low ambient temperatures and during the following recoveries at ambient temperature 24 degrees C. DESIGN: Electroencephalographic activity, hypothalamic temperature, and motor activity were studied during a 24-hour exposure to ambient temperatures ranging from 10 degrees C to -10 degrees C and for 4 days during the recovery. SETTING: Laboratory of Physiological Regulation during the Wake-Sleep Cycle, Department of Human and General Physiology, Alma Mater Studiorum-University of Bologna. SUBJECTS: Twenty-four male albino rats. INTERVENTIONS: Animals were implanted with electrodes for electroencephalographic recording and a thermistor for measuring hypothalamic temperature. MEASUREMENTS AND RESULTS: Wake-sleep stage duration and the electroencephalographic spectral analysis performed by fast Fourier transform were compared among baseline, exposure, and recovery conditions. The amount of non-rapid eye movement sleep was slightly depressed by cold exposure, but no rebound was observed during the recovery period. Delta power during non-rapid eye movement sleep was decreased in animals exposed to the lowest ambient temperatures and increased during the first day of the recovery. In contrast, rapid eye movement sleep was greatly depressed by cold exposure and showed an increase during the recovery. Both of these effects were dependent on the ambient temperature of the exposure. Moreover, theta power was increased during rapid eye movement sleep in both the exposure and the first day of the recovery. CONCLUSION: These findings show that sleep-stage duration and electroencephalogram power are simultaneously affected by cold exposure. The effects on rapid eye movement sleep appear mainly as changes in the duration, whereas those on non-rapid eye movement sleep are shown by changes in delta power. These effects are temperature dependent, and the decrease of both parameters during the exposure is reciprocated by an increase in the subsequent recovery. PMID- 16477957 TI - Impairment of error monitoring following sleep deprivation. AB - STUDY OBJECTIVES: To verify if error monitoring, involving detection and remedial actions, is affected by sleep deprivation. DESIGN: Event-related brain potentials (ERPs) and electroencephalogram spectrum during performance of Flanker task were obtained in a within-subject, counter-balanced, repeated-measures design. SETTING: Sleep deprivation and data collection were conducted in a laboratory setting. PARTICIPANTS: Sixteen young healthy adults (7 women, 18-23 years old) INTERVENTIONS: Performance and electroencephalogram data were collected after normal sleep and sleep deprivation. MEASUREMENTS AND RESULTS: Compared to normal sleep, 1 night of sleep deprivation resulted in slower and more varied reaction times, more response errors and omissions, and impaired posterror adjustments to response accuracy. Concomitantly, 2 error-related ERPs, error-related negativity and Pe, showed reduced amplitude measurements after sleep deprivation. Conversely, conflict monitoring as expressed behaviorally and by the N2 component of the ERP was not attenuated by sleep deprivation. Ten of the sixteen participants maintained similar accuracy levels under both sleep conditions, although they still showed reduced error-related negativity and error positivity amplitude measurements and impaired error remedial actions for accuracy. Electroencephalogram spectral activity at beta and theta frequency bands was related to response correctness on subsequent trials but not related to that of preceding trials under both sleep conditions. CONCLUSIONS: One night of sleep deprivation impaired both the error detection and error remedial actions and highlighted the inability to avoid making errors again after erroneous responses were already made. The results showed that a vicious cycle occurred between performance deterioration and impairment of error-remedial mechanisms that inevitably led to making more successive errors. PMID- 16477958 TI - Physician and patient determinants of pharmacologic treatment of sleep difficulties in outpatient settings in the United States. AB - STUDY OBJECTIVES: This study analyzed patient and physician characteristics associated with physicians' choice of medication treatment for sleep difficulties in a nationally representative sample of outpatient physician visits in the United States. DESIGN: Cross-sectional study of nationally representative survey data. SETTING: Outpatient settings in the United States. PATIENTS OR PARTICIPANTS: Patients aged 18 years or older with sleep difficulty. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: The National Ambulatory Medical Care Survey data from 1996 to 2001 were utilized. The study found that approximately 4.8 billion visits were made to outpatient physician offices in the United States, and 94.6 million of these were visits related to sleep difficulties. Visits by established patients, compared with visits by new patients, were associated with an increased prescription of some type of pharmacotherapy (odds ratio: 1.92, 95% confidence interval: 1.35-2.73). Patient visits with public insurance, as compared with private insurance, as a primary payer source were associated with increased benzodiazepine prescriptions among patients receiving pharmacotherapy (odds ratio: 1.66, 95% confidence interval: 1.13-2.45). Visits by patients aged 65 years and older, as compared with those by patients aged 18 to 34 (reference group)., had almost two times increased odds of being associated with a benzodiazepine prescription (odds ratio: 1.89, 95% confidence interval: 1.13-3.14) among patient visits receiving pharmacotherapy. CONCLUSIONS: The results of this study indicate that several patient and physician characteristics influence physician prescribing of pharmacologic treatments for sleep difficulties. The study also finds variations in pharmaceutical treatment for sleep difficulties in outpatient settings in the United States. PMID- 16477959 TI - A 2-week efficacy and safety study of eszopiclone in elderly patients with primary insomnia. AB - STUDY OBJECTIVES: Evaluate the efficacy of eszopiclone in primary insomnia. DESIGN/SETTING: Randomized, double-blind, placebo-controlled multicenter in outpatient setting with weekly visits. PARTICIPANTS: Two-hundred thirty one men and women aged 65 to 85 years (mean age 72.3 years) with primary insomnia, as defined by the Diagnostic and Statistical Manual of Mental Disorders-Fourth Edition. INTERVENTIONS: Eszopiclone 1 mg (n = 72), eszopiclone 2 mg (n = 79), or placebo (n = 80) nightly for 2 weeks. MEASUREMENTS/RESULTS: Efficacy was assessed using an interactive voice response system. Following the predefined hierarchical testing strategy, the eszopiclone 2-mg group had a significantly shorter sleep latency compared with placebo over the double-blind period (P = .0034). The eszopiclone 2-mg group had significantly longer total sleep time (P = .0003) and eszopiclone 1-mg group had significantly shorter sleep latency (P < or = .012) compared with placebo. The eszopiclone 1-mg group was not significantly different from placebo on total sleep time or any other secondary efficacy endpoint. Secondary analyses indicated that the eszopiclone 2-mg group had significantly less wake after sleep onset; significantly fewer and shorter in duration daytime naps; and significantly higher ratings of sleep quality and depth, daytime alertness, and sense of physical well-being compared with placebo (P < .05). Eszopiclone was well tolerated. The most frequent treatment-related adverse event was unpleasant taste. CONCLUSION: Nightly treatment with eszopiclone 1 mg effectively induced sleep, while the 2-mg dose was effective in inducing and maintaining sleep. Eszopiclone was well tolerated in elderly patients with primary insomnia, and the sleep efficacy was accompanied by significantly less napping and significantly higher ratings of daytime alertness, sense of physical well-being, and several quality-of-life parameters at the higher dose. PMID- 16477960 TI - Relationships of occupational stress to insomnia and short sleep in Japanese workers. AB - STUDY OBJECTIVE: The aims of this study were to (a) examine the association between occupational stress and insomnia and short sleep in Japanese workers and (b) demonstrate the difference between 2 occupational stress models-Effort Reward Imbalance and the Demand Control Model. DESIGN: All data were obtained via self administrated questionnaires and annual health checkups. Insomnia was evaluated by the Athens Insomnia Scale, and short sleep was defined as less than 6 hours sleep per day. SETTING: Employees at local governments and a transit company who had annual health checkups during the period from April 2003 to March 2004. PARTICIPANTS: After excluding participants without complete data, data from 6,997 men and 1,773 women were analyzed. INTERVENTION: N/A. MEASUREMENTS AND RESULTS: In men, high occupational stresses were significantly associated with insomnia, especially a high level of Effort Reward Imbalance (defined as the presence of high effort and low reward), had a remarkably higher odds ratio. In women, high occupational stresses were significantly associated with insomnia as well. High occupational stresses were significantly associated with short sleep in men. However, in women, only Effort Reward Imbalance showed a significant association with short sleep. CONCLUSIONS: This study suggested that occupational stress is a possible risk factor for insomnia and short sleep. PMID- 16477961 TI - Severity of sleep-disordered breathing improves following parturition. AB - STUDY OBJECTIVE: Changes in sleep-disordered breathing associated with late pregnancy have not previously been systematically investigated; however, a number of case reports indicate exacerbation of obstructive sleep apnea in late pregnancy, often in association with maternal hypertension. We aimed to compare the severity of sleep-disordered breathing and associated maternal blood-pressure responses in late pregnancy with the nonpregnant state. DESIGN: Case-controlled, longitudinal study of sleep-disordered breathing during late pregnancy and postpartum. STUDY PATIENTS: Ten women referred for suspected sleep-disordered breathing during the third trimester of pregnancy. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Full overnight polysomnography and continuous systemic blood pressure were measured during the third trimester of pregnancy and 3 months following delivery. Parameters of sleep-disordered breathing, including apnea hypopnea index and minimum overnight arterial oxyhemoglobin saturation, were compared between antenatal and postnatal studies. An improvement in both apnea hypopnea index and minimum arterial oxyhemoglobin saturation occurred consistently in all subjects postnatally. In non-rapid eye movement sleep, mean apnea-hypopnea index was reduced from 63 +/- 15 per hour antenatally to 18 +/- 4 per hour postnatally (P = .03), and in rapid eye movement sleep, from 64 +/- 11 per hour to 22 +/- 4 per hour (P = .002). Minimum arterial oxyhemoglobin saturation was increased from 86% +/- 2% antenatally to 91% +/- 1% postnatally (P = .01). Arterial blood-pressure responses to apnea peaked at 170 to 180 mm Hg antenatally, while they only peaked at 130 to 140 mm Hg postnatally. CONCLUSION: This study indicates that late pregnancy may be associated with increased severity of sleep-disordered breathing and associated blood-pressure responses. PMID- 16477962 TI - Autonomic dysfunction in children with sleep-disordered breathing. AB - STUDY OBJECTIVES: To measure sympathetic responses in children with and without sleep-disordered breathing. DESIGN: Prospective, observational study. SETTING: Kosair Children's Hospital Sleep Medicine and Apnea Center. PARTICIPANTS: Subjects were prospectively recruited from children undergoing overnight polysomnographic assessments and were retrospectively grouped according to the results of the polysomnogram. Sleep-disordered breathing was defined as an apnea hypopnea index >5 and children were assigned to the control group if their apnea hypopnea index was < 1. INTERVENTION: N/A. MEASUREMENTS AND RESULTS: During quiet wakefulness, pulse arterial tonometry was used to assess changes in sympathetic activity following vital capacity sighs in 28 children with sleep-disordered breathing and 29 controls. Each child underwent a series of 3 sighs, and the average maximal pulse arterial tonometry signal attenuation was calculated. Further, a cold pressor test was conducted in a subset of 14 children with sleep disordered breathing and 14 controls. The left hand was immersed in ice cold water for 30 seconds while right-hand pulse arterial tonometry signal was continuously monitored during immersion and 20-minute recovery periods. Signal amplitude changes were expressed as percentage change from corresponding baseline. RESULTS: The magnitude of sympathetic discharge-induced attenuation of pulse arterial tonometry signal was significantly increased in children with sleep-disordered breathing during sigh maneuvers (74.1% +/- 10.7% change compared with 59.2% +/- 13.2% change in controls; P<.0001) and the cold pressor test (83.5% +/- 7.3% change compared with 74.1% +/- 11.4% change in controls; P=.039). Further, recovery kinetics in control children were faster than those of children with sleep-disordered breathing. CONCLUSION: Children with sleep-disordered breathing have altered autonomic nervous system regulation as evidenced by increased sympathetic vascular reactivity during wakefulness. PMID- 16477963 TI - Emerging therapies in narcolepsy-cataplexy. AB - In the past, narcolepsy was primarily treated using amphetamine-like stimulants and tricyclic antidepressants. Newer and novel agents, such as the wake-promoting compound modafinil and more selective reuptake inhibitors targeting the adrenergic, dopaminergic, and/or serotoninergic reuptake sites (ie, venlafaxine, atomoxetine) are better-tolerated available alternatives. The development of these agents, together with sodium oxybate (a slow-wave sleep-enhancing agent that consolidates nocturnal sleep, reduces cataplexy, and improves sleepiness), has led to improved functioning and quality of life for many patients with the disorder. However, these treatments are all symptomatically based and do not target hypocretin, a major neurotransmitter involved in the pathophysiology of narcolepsy. In this review, we discuss emerging therapies in the area of narcolepsy. These include novel antidepressant or anticataplectic, wake promoting, and hypnotic compounds. We also report on novel strategies designed to compensate for hypocretin deficiency and on the use of immunosupression at the time of narcolepsy onset. PMID- 16477964 TI - Mathematical properties of the sleep pressure score. PMID- 16477965 TI - An integrated electro-chemical and natural treatment system for industrial water pollution control. AB - Experiments were conducted to test the feasibility of applying an integrated electro-chemical (EC) and natural treatment system for treatment of some industrial wastewaters. The EC process was found to be very effective in removing lead, a model heavy metal from some wastewaters. Within 20 minutes of operation time, 5 to 10 A of electric current and specific surface area of electrode of 46.51 m2/m3, the lead concentrations in the wastewaters were reduced from 35-100 mg/l to less than 1 mg/l. Based on a kinetic model developed from the experimental data, the important parameters for the EC process were found to be electric current, specific surface area of electrode, and operation time. From scanning electron microscopic and X-ray diffractometric (XRD) analysis, the EC sludge samples were found to compose mainly of maghemite (Fe2O3), magnetite (Fe3O4), and laurionite (PbClOH), suitable for disposal to secure landfills. Two pilot-scale constructed wetlands (CW) in series, a model natural treatment system, were employed to treat wastewaters of an industrial estate in Thailand. At organic loading rates of 57-140 kg BOD/hectare-year, these constructed wetlands were able to reduce BOD from 90 to 4 mg/l, while suspended solids, total nitrogen and total phosphorus were reduced from 100 to 10 mg/l, 24 to 4.6 mg/l and 7 to 1.5 mg/l, respectively, during the summer season. These results demonstrated technical feasibility of CW in removing organic and other pollutants contained in this industrial wastewater. PMID- 16477966 TI - Bioflocculent algal-bacterial biomass improves low-cost wastewater treatment. AB - An innovative technology for the biological treatment of wastewater in regions with sufficient solar radiation based on the simultaneous growth and degradation processes of algal and bacterial biomass is presented. The aim of the work is the improvement of pond technology through the formation of stable algae-bacteria aggregates, which a) permit a simple separation of the algal biomass by gravity sedimentation, b) enable a high removal efficiency for organic carbon and nutrients, and c) are independent in terms of oxygen provision through algal photosynthesis. Algae-bacteria aggregates could be developed with a suitable algal species (Chlorella vulgaris, Strain Hamburg) as a 'model organism' in a wastewater environment. The morphology of algal-bacterial flocs is similar to activated sludge flocs. They are stable and settle quickly. Floc size ranged between 400 and 800 microm. Results of our experiments with an artificially irradiated lab-scale system, operated in continuous flow mode, revealed that even at a relatively short hydraulic detention time of two days, a high elimination capacity of 9.96 g N m(-2) d(-1) and 0.87g Pm(-2) d(-1) can be achieved. Recent investigations confirmed that floc formation of unicellular algae and wastewater bacteria also could be developed and maintained in a pilot-scale system with a water depth of 0.5 m. PMID- 16477967 TI - A sustainable approach towards rural development: dry toilets in Nepal. AB - Existing inadequate sewerage systems and direct disposal of household waste into water courses has tremendously increased water pollution. Dry toilets are feasible in rural and peri-urban areas to reduce the consumption of costlier water that is required for flushing. As conventional treatment technologies require high investment, and operation and maintenance costs, dry toilets are the only suitable option left for sanitation in the 21st century when working with limited financial resources. To reduce environmental degradation and overcome this problem, the dry toilet is only the realistic option in Nepal. Two reactors, one exposed to sunrays and the other without sunrays, were constructed. In the model with sunrays, it was found that in 48 days of observation faecal coliform presence depleted to 610 cells per gm from the initial value of 7 x 10(10) and volatile organic matter came down from 98.09% to 70.18%. Similarly, in the other model, the destruction of faecal coliform in 65 days was found to be 920 cells/gm while the destruction of organic matter took 75 days. Also, observing from 313 people in a cluster on the pilot project, the annually recovered value of N, P and K was found to be 1565 kg, 125 kg, 344 kg, respectively. This paper deals with the different types of dry ecological toilet, their performance and feasibility study in Nepal, with the full involvement of local people, based on complete laboratory analysis and regular monitoring. Using dry toilets will save 14 LPCD, which is equivalent to 14 MLD and the resulting demand will become only 80 MLD for the urban area of Kathmandu. The result advocates the implementation of ecological dry toilets to save valuable water wasted in flushing, as well as saving the resources needed to treat the waste. PMID- 16477968 TI - Sustainable sewage solutions for small agglomerations. AB - In a significant number of European countries, the need for providing appropriate treatment for the effluents of small rural communities is still especially relevant. In fact, in countries like Portugal, Spain, and many others, significant amounts of investment will be addressed in the next few years to the construction of small Wastewater Treatment Plants (WWTP). The problems faced when constructing and operating WWTP serving small communities may be relevant when energy and labour costs are relatively high, or when the visual impact on the surrounding areas is especially negative. Sustainable treatment solutions require the selection of appropriate technologies using fewer resources. In this paper, information is presented about sustainability indicators of twenty-one small secondary wastewater treatment plants, including conventional (trickling filters and extended aeration plants) and non-conventional treatment systems (constructed wetlands). The data refer to allocated areas per inhabitant, amounts of concrete per inhabitant, power per inhabitant, and construction and installation costs per inhabitant. The data seem to show that for different reasons, constructed wetlands are promising treatment solutions for application to rural areas in particular because of the relatively low power requirements and relatively low construction costs for served populations below 500 inhabitants. PMID- 16477969 TI - Preparing public participation at the catchment level: comparison of three methodologies applied to the Herault river basin. AB - Public participation in water resources management has increasingly been recommended to increase the effectiveness and the acceptability of proposed water management projects and plans. Different levels of public participation are possible depending on the governance context. This paper reports on the practical implementation of three different methodologies in the Herault river catchment (France). The analysis of public view points, carried out using a quantitative opinion survey and focus groups, reveals important information needs which have to be fulfilled for any further consultation to be efficient. A complementary analysis of stakeholders' opinions, carried out through semi-structured interviews also highlights the need to construct a common knowledge base between stakeholders, expert and scientists. This is seen as a prerequisite to implementing more elaborated forms of participation. PMID- 16477970 TI - Urban water tariff structure and cost recovery opportunities in India. AB - Given the high level of capital investment and the history of government subsidized services, full cost pricing of water services has yet to take hold in India. As a result, it remains broadly underpriced leading to public perception that water is "free" The current tariff levels in India are too low to cover even operating costs. This paper examines the existing Indian urban water tariff models (fixed tariff, volumetric tariff, increasing block tariff etc.), their relevance and problems. It was found that none of the tariff structures could satisfy all the design objectives (cost recovery, economic efficiency, equity, affordability etc.). Also subsidies are not explicit and well targeted for poor population. There are several studies and issues that do demonstrate the opportunities for tariff increase and improved cost recovery. This paper highlights the results of such studies and brings out issues needing consideration. Improved cost recovery would lead to improved financial status of the water utilities. Also, subsidies, if designed suitably and well targeted, would serve the concerns of the economically weaker sections. Such reform process would eventually lead to socio-economic sustainability. PMID- 16477971 TI - Risk management in waste water treatment. AB - With the continuous restructuring of the water market due to liberalisation, privatisation and internationalisation processes, the requirements on waste water disposal companies have grown. Increasing competition requires a target-oriented and clearly structured procedure. At the same time it is necessary to meet the environment-relevant legal requirements and to design the processes to be environment-oriented. The implementation of risk management and the integration of such a management instrument in an existing system in addition to the use of modern technologies and procedures can help to make the operation of the waste water treatment safer and consequently strengthen market position. The risk management process consists of three phases, risk identification, risk analysis/risk assessment and risk handling, which are based on each other, as well as of the risk managing. To achieve an identification of the risks as complete as possible, a subdivision of the kind of risks (e.g. legal, financial, market, operational) is suggested. One possibility to assess risks is the portfolio method which offers clear representation. It allows a division of the risks into classes showing which areas need handling. The determination of the appropriate measures to handle a risk (e.g. avoidance, reduction, shift) is included in the concluding third phase. Different strategies can be applied here. On the one hand, the cause-oriented strategy, aiming at preventive measures which aim to reduce the probability of occurrence of a risk (e.g. creation of redundancy, systems with low susceptibility to malfunction). On the other hand, the effect-oriented strategy, aiming to minimise the level of damage in case of an undesired occurrence (e.g. use of alarm systems, insurance cover). PMID- 16477972 TI - Planning, modelling and assessing source control concepts on catchment scale. AB - Today's water management practice was identified often not to be consistently structured. Usually the decision space is not clearly depicted and then systematically explored. Often stakeholders are involved too late and many objectives are neglected. An adaptive DSS is being developed to help overcoming these problems. Core of the DSS is a "decision matrix", which has been implemented as a web based application (www.wsm300.de). The management objectives are represented by the indicators labelling the rows. They will be the result of a discussion of the objectives and problems in the specific sub-basin, which is supported by the catalogue of indicators. The columns of the matrix are specified by the scenarios whose development is supported by the database of measures and a Geographical Information System (GIS). The matrix, once the labels are defined, serves as a plot for the planning process, defining clearly which objectives have to be considered and which indicator-values have to be calculated. The DSS further includes a concept and tools for the combination of existing software components and supports the processing of model-outputs to indicator-values. Filled with the indicator-values the matrix allows a comparison of the scenarios and provides a good base for a decision. Multi-criteria decision aid methods can (if desired) further help to find the optimal scenario and to mediate between stakeholders. Finding the optimal scenario will likely be an iterative process. PMID- 16477973 TI - Integrated modelling of conventional pollutants and organic contaminant fate in rivers: a microcosm study. AB - A new conceptual dynamic integrated model is presented which can be used to describe both conventional pollutants and organic contaminant fate in rivers. The model is designed to assess the short-term fate of organic contaminants in two compartments (bulk water and benthic sediment), taking into account the effect of nutrient dynamics. The biodegradation submodel was refined using a microcosm (artificial river) study and Linear Alkylbenzene Sulphonate (LAS) as an example. Based on data generated during the microcosm study, the model was calibrated and validated in both steady state (continuous constant load) and dynamic (pulse load) conditions. The results show that the simulated data set agrees well with the measured data set. Furthermore, thorough investigation of the model output sensitivity to the model inputs was made, and the results show that the fate of LAS is sensitive to the following model input variables: ammonia nitrogen, dissolved oxygen, microbial biomass and readily biodegradable soluble COD, and the model parameters mainly related to the biodegradation submodel. The model provides good understanding of the interaction between conventional pollutants and organic contaminants fate in rivers. PMID- 16477974 TI - Development of water quality models for supporting NH3-N control in a dam regulated river. AB - A seasonal occurrence of high ammonia nitrogen (NH3-N) concentrations has hampered chemical treatment processes of a water plant in Geum river of Korea. Monthly flow allocation from upstream dam is important for downstream NH3-N control. In this study, water quality models based on multiple regression (MR) and artificial neural network (ANN) methods were developed to support dam operations through providing forecasted NH3-N concentrations. The models were calibrated with 7 years of monthly data, and verified with another 3 years of independent data. In the models, the NH3-N concentration for next time step is dependent on dam outflow, river water quality such as alkalinity, temperature, and NH3-N of previous time step. During the calibration phase, the ANN models compared to MR models showed a better agreement with observed data as indicated with small RMSE and high R2 values. However, in the verification phase, the performance of ANN models was decreased and showed a little difference to MR models. From the model comparisons, it is recommended for both ANN and MR models to include the autocorrelation of NH3-N concentrations up to 3-lag months to avoid overestimation during low flow season. PMID- 16477975 TI - UVQ: a tool for assessing the water and contaminant balance impacts of urban development scenarios. AB - This paper presents the water and contaminant daily simulation model of the total water cycle, called UVQ. The model has been developed to provide a means for rapidly assessing conventional and nonconventional approaches to providing water supply, stormwater and wastewater services to urban allotments, neighbourhoods and study areas. The model is placed the context of other such models developed internationally through a brief literature review. This is followed by a description of the model and output examples, which is used to illustrate the utility of the model. UVQ is an effective preliminary assessment tool for determining the impacts of urban development options on the total water cycle, as well as the performance of a wide range of non-conventional demand and supply side management techniques. It compliments other aspects of an environmental assessment of options, along with more traditional aspects such as infrastructure costing. PMID- 16477976 TI - Computational intelligence-based optimisation of wastewater treatment plants. AB - Methods of computational intelligence (CI), especially fuzzy control and neuronal networks, are used for controlling and optimising of wastewater treatment plants. Areas of application are the control of sludge water dosage, of phosphate elimination by optimal precipitant dosage as well as an optimal aeration in the nitrification zone. In two municipal wastewater treatment plants with 60,000 and 12,600 person equivalents the controllers have been installed and optimised and they have been in operation for several years. Results of operation of the plants are presented in comparison to previously used classical control. Performance increased significantly and the outflow values could be kept securely below the government requirements without increase of the energy consumption. Peak loads in the inflow were eliminated in the plant and did not increase outflow concentrations. Results of operation for more than three years clearly show that the CI controller is a cost-efficient method for a sustainable rise of performance in municipal wastewater treatment plants. PMID- 16477977 TI - Granulation in an upflow anaerobic sequencing batch reactor treating disintegrated waste activated sludge. AB - An upflow anaerobic reactor operated with a sequencing batch mode to enhance high rate digestion of raw and thermally disintegrated waste activated sludge with formation of granules. The gas production rate doubled when disintegrated waste activated sludge was introduced. Gradual granulation took place and the dispersed particles become coarse granulation as the operation continued. The granular sludge showed relatively higher specific methanogenic activity than the dispersed sludge. Bacterial morphology by a scanning electron microscope showed diversity of bacteria such as filamentous, rod and spherical shape in the section of granules. Filamentous bacteria, which might support the frame of a granule, were observed as long chains at the outer surface. Meanwhile, rod and spherical bacteria, which might play a role in the initial stage of granule formation, were observed from the inner surface of the granule. High rate digestion of sludge along with efficient liquid-solids separation was achieved due mainly to development of sludge granules within the upflow reactor. PMID- 16477978 TI - A benchmark study of controlled emptying of equalization basins. AB - Storm tanks, or equalization basins, have been used for many years in sewer or wastewater treatment systems to reduce the amount of combined sewer overflows. In this paper, a systematic control of an equalization basin is used not only to dampen or reduce the hydraulic load, but also to systematically improve the effluent quality in a nitrogen removal plant. It is demonstrated that the effluent ammonia concentration is the key component. The Benchmark Simulation Model 1, implemented in MATLAB/Simulink, has been used to perform the evaluations. It has been extended with systematic rainfall generations. Intuitively, it is apparent that the equalization basin should be emptied before a rainfall. This requires a prediction capacity. It is shown that the choice of prediction time is by no means trivial and it is not true that a long prediction horizon is always advantageous. PMID- 16477979 TI - Setting-up a cost recovery system for the largest wastewater treatment plant in South-East Asia. AB - A tariff system has been set up for the largest wastewater treatment plant in South-East Asia, the Samut Prakarn Wastewater Treatment Plant south of Bangkok, which is currently under completion. Fully functional the plant will have a design capacity for 500,000 m3 per day and will service a combined residential and industrial area with approximately 600,000 residents and 2,300 factories. The tariff system, which includes a tariff model, is based on water consumption and BOD load. As background for setting the tariffs a comprehensive monitoring system including an industrial permitting system has been developed. The paper presents the background and rationale for setting up the system as well as the objective, scope and content of the tariff system and the industrial permit system. Further, the feasibility of introducing cost recovery systems, which is widely accepted in developing economies on the conceptual level and to some extent implemented at the legal and regulatory level, but has yet to be implemented at large, is discussed. PMID- 16477980 TI - Technical-economical evaluation of the operation of oxidation ditches. AB - The optimisation of the economic performance is a central aspect in the management of wastewater treatment plants. A model-based procedure was developed that could provide a clearly enunciated and concise way of reporting to the stakeholders on how well the assets are performing and can perform, separating the firm inefficiency from the effect of the treatment. The applied modelling method is conventional considering current modelling research practice, but very good from practical point of view in that it is applicable utilising existing plant data, i.e. without the use of additional measuring campaigns. This paper illustrates the validity of the developed procedures through the evaluation of the performance of oxidation ditches. PMID- 16477981 TI - Potential of OUR and OTR measurements for identification of activated sludge removal processes in aerated basins. AB - In order to develop a process control scheme to reduce energy costs for aeration in activated sludge systems with biological P removal, pre-denitrification and nitrification stages, the spatial distribution of carbon oxidation and nitrification was evaluated over a long full-scale plug flow aeration basin using an externally measured specific oxygen uptake rate (sOUR) and in basin measurement of the actual specific oxygen transfer rate (sOTR) with off-gas testing as well as with the calculated oxygen demand from NH4-N concentrations (sOTR(N)). Using a simple static model, a gas phase balance on oxygen and carbon dioxide, sOTR(N) values were also calculated from off-gas testing. Comparison of sOTR(N) to sOTR and sOUR for carbon oxidation (sOUR(C)) to nitrification (sOUR(N)) at different loading conditions allowed the oxidation processes to be followed over the three zones of the aeration basin. As expected, the distribution depended on the dissolved oxygen concentration (DO) in the basin. However, the major change was in the C-oxidation rate and not the nitrification rate. At a low DO, and when NH4-N was present in the zone, the amount of oxygen transferred for nitrification was nearly the same, but the overall sOTR was lower. The externally measured sOUR was only useful when it was differentiated into sOUR(N) and sOUR(C). sOUR(N) could be used to predict the nitrification rate in the basin. With further refinement, the gas phase balance model has potential to be used to monitor the degree of nitrification over the basin length. This can be integrated into a control scheme to reduce aeration costs by adjusting the DO setpoint according to loading conditions in the PMID- 16477982 TI - Experience from 10 years of full-scale operation with enhanced biological phosphorus removal at Oresundsverket. AB - Ten years of full-scale experience with enhanced biological phosphorus removal (EBPR) has been evaluated. During the start-up period lack of carbon source was the main operational problem and a higher level of volatile fatty acids was secured by introducing a primary sludge hydrolysis. Acidic thermal sludge hydrolysis was used as the sludge treatment method at the plant during about three years. One effluent stream, rich in carbon and precipitant, was brought back to the process leading to an improvement of the phosphorus removal both by an improved biological process and chemical precipitation. A quite stable process of EBPR was developed with low levels of effluent phosphorus concentration. Stringent effluent discharge limits during short evaluation periods necessitated a continued work for improvement of the short-term stability. During periods with lack of carbon, such as industrial holiday or rainy periods, both simultaneous precipitation and reduced aeration have been successfully tested as strategies for securing low levels of effluent phosphorus. PMID- 16477984 TI - Systems analysis of urban wastewater systems--two systematic approaches to analyse a complex system. AB - This work was aimed at performing an analysis of the integrated urban wastewater system (catchment area, sewer, WWTP, receiving water). It focused on analysing the substance fluxes going through the system to identify critical pathways of pollution, as well as assessing the effectiveness of energy consumption and operational/capital costs. Two different approaches were adopted in the study to analyse urban wastewater systems of diverse characteristics. In the first approach a wide ranged analysis of a system at river basin scale is applied. The Nete river basin in Belgium, a tributary of the Schelde, was analysed through the 29 sewer catchments constituting the basin. In the second approach a more detailed methodology was developed to separately analyse two urban wastewater systems situated within the Ruhr basin (Germany) on a river stretch scale. The paper mainly focuses on the description of the method applied. Only the most important results are presented. The main outcomes of these studies are: the identification of stressors on the receiving water bodies, an extensive benchmarking of wastewater systems, and the evidence of the scale dependency of results in such studies. PMID- 16477983 TI - Fuzzy control of nitrogen removal in predenitrification process using ORP. AB - In order to meet increasingly stringent discharge standards, new applications and control strategies for the sustainable removal of nitrogen from wastewater have to be implemented. In the past years, numerous studies have been carried out dealing with the application of fuzzy logic to improve the control of the activated sludge process. In this paper, fuzzy control strategies of predenitrification systems are presented that could lead to better effluent quality and, in parallel, to a reduction of chemicals consumption. Extensive experimental investigations on lab scale plant studies have shown that there was excellent correlation between nitrate concentration and ORP value at the end of the anoxic zone. Results indicated that ORP could be used as an on-line fuzzy control parameter of nitrate recirculation and external carbon addition. The optimal value of ORP to control nitrate recirculation and external carbon addition was - 86 +/- 2 mV and - 90 +/- 2 mV, respectively. The results obtained with real wastewater also showed the good performance and stability of the fuzzy controllers independently from external disturbances. The integrated control structure of nitrate recirculation and external carbon addition in the predenitrification system is also presented. PMID- 16477985 TI - Current state and development of the real-time control of the Berlin sewage system. AB - Since the 1970s, we have known about real-time control of urban drainage systems. However, global real-time control strategies still show a lack of implementation for large drainage systems of high complexity. In Berlin, Germany, a city of 3.5 million inhabitants covering an area of around 900 km2, the demand for enhanced protection of the environment and growing economic pressure have led to an increasing application of control assets and concepts within the sewage system. In the framework of the project "Integrated Sewage Management", the possibilities of a global and integrated control strategy for the Berlin system are examined. The paper is focused on the historical concept and design of the sewerage and the further improvement towards an environment-oriented system that builds the basis for today's considerations. The operational method and functionality of local regulators that have already been implemented are described. Further-more, the model-based methodology for the analysis of the system and the development of global control concepts, as well as the results of system analysis, are stated. On the basis of model simulations, it is shown that a global coordination of pump stations can lead to a reduction of sewer overflows, and consequently to an enhanced water protection. PMID- 16477986 TI - Sustainable management of leakage from wastewater pipelines. AB - Wastewater pipeline leakage is an emerging concern in Europe, especially with regards to the potential effect of leaking effluent on groundwater contamination and the effects infiltration has on the management of sewer reticulation systems. This paper describes efforts by Australia, in association with several European partners, towards the development of decision support tools to prioritize proactive rehabilitation of wastewater pipe networks to account for leakage. In the fundamental models for the decision support system, leakage is viewed as a function of pipeline system deterioration. The models rely on soil type identification across the service area to determine the aggressiveness of the pipe environment and for division of the area into zones based on pipe properties and operational conditions. By understanding the interaction between pipe materials, operating conditions, and the pipe environment in the mechanisms leading to pipe deterioration, the models allow the prediction of leakage rates in different zones across a network. The decision support system utilizes these models to predict the condition of pipes in individual zones, and to optimize the utilization of rehabilitation resources by targeting the areas with the highest leakage rates. PMID- 16477987 TI - Benchmarking management of sewer systems: more to learn than cost effectiveness. AB - Thirty-nine municipalities in the Netherlands conducted a pilot study to develop and try out a methodology to compare the quality of their sewerage management. The participants chose a multidimensional benchmarking with an emphasis on the aim of improving the working processes within sewerage management. A second goal was accountability to the stakeholders. The benchmarking methodology was based as well on analysing data within a "balanced-score-card" system as on intensive exchange of knowledge and experiences. The pilot resulted in a state of the art overview of the quality of sewerage management in the Netherlands. However, above all, it resulted in the shocking fact that the work is carried out in many different ways which cannot be explained by technical reasons or local circumstances. To pinpoint best practices and actually implement these improvements the learning process must continue after the analysis and presentation of the data. A start has been made to form regional specialist networks for further discussion and exchange of experiences. PMID- 16477988 TI - The hydraulic capacity of deteriorating sewer systems. AB - Sewer and wastewater systems suffer from insufficient capacity, construction flaws and pipe deterioration. Consequences are structural failures, local floods, surface erosion and pollution of receiving waters bodies. European cities spend in the order of five billion Euro per year for wastewater network rehabilitation. This amount is estimated to increase due to network ageing. The project CARE-S (Computer Aided RE-habilitation of Sewer Networks) deals with sewer and storm water networks. The final project goal is to develop integrated software, which provides the most cost-efficient system of maintenance, repair and rehabilitation of sewer networks. Decisions on investments in rehabilitation often have to be made with uncertain information about the structural condition and the hydraulic performance of a sewer system. Because of this, decision-making involves considerable risks. This paper presents the results of research focused on the study of hydraulic effects caused by failures due to temporal decline of sewer systems. Hydraulic simulations are usually carried out by running commercial models that apply, as input, default values of parameters that strongly influence results. Using CCTV inspections information as dataset to catalogue principal types of failures affecting pipes, a 3D model was used to evaluate their hydraulic consequences. The translation of failures effects in parameters values producing the same hydraulic conditions caused by failures was carried out through the comparison of laboratory experiences and 3D simulations results. Those parameters could be the input of 1D commercial models instead of the default values commonly inserted. PMID- 16477989 TI - Continuous monitoring in sewer networks an approach for quantification of pollution loads from CSOs into surface water bodies. AB - Different approaches for quantification of pollution loads discharged from combined sewer networks into surface water bodies have been observed over the last few years and decades, but a large number of unresolved problems still remain. Many monitoring campaigns have been based on manual or automated spot sampling - with the long known limitations of this method such as sampling errors and errors due to sample conservation, transport and preparation. On the other hand, only recently have sensors became available which are suitable for continuous application in sewer networks. A large number of practical problems still have to be solved before continuous monitoring in sewer networks will be successful. Additionally, most of the applicable sensors are based on surrogate methods which results in a considerable effort for reference measurements for sensor calibration. Finally, it has to be considered that, depending on the sewer network topography, deposition and remobilisation of pollutants varies considerably, which limits the generality of monitoring results and, subsequently, their applicability as a base for the design of storm water tanks or combined sewer overflows (CSO). A monitoring station for continuous monitoring of load discharges from a CSO has been installed and operated for more than one year. The design and equipment of the measurement station, operational experiences and results are given in this paper. PMID- 16477990 TI - A new installation for treatment of road runoff: up-flow filtration by porous polypropylene media. AB - We installed a new device on a paved road to treat runoff from a roadway surface. All the stormwater runoff was transferred into the device and the runoff equivalent to 10 mm/hr or less was treated. The treatment method consists of sedimentation and up-flow filtration with porous polypropylene (PPL) processes. The treated runoff was discharged into the existing storm drainage pipe. The average removal efficiency of the initial runoff at the beginning of rainfall which has high pollution intensity was about 90% for SS, about 70% for COD, about 40% for total phosphorus (T-P), about 80% for Pb and Cd, about 70% for Zn, Cu, Mn and Cr, and about 60% for polycyclic aromatic hydrocarbons (PAHs). The overall removal efficiencies of the experiment that ran for four months remained > 60% of SS, > 40% of COD, > 60% of heavy metals, and > 40% of PAHs. The PPL is excellent for removing smaller size particulates of suspended solids, which originate basically from diesel exhaust, as well as larger size particulates from automobile tires, asphalt roads, and other accumulated source(s) of clay and sand, etc. PMID- 16477991 TI - Dissolved organic matter from agricultural fields in the irrigation period. AB - The aim of this study was to quantify and characterize the dissolved organic carbon (DOC) of paddy fields and crop fields in Tottori, Japan. Dissolved organic carbon (DOC) and ultraviolet (UV) absorbance was measured for the filtrated water of each samples. DOC concentration and SUVA (specific UV absorbance) of biodegradation analysis samples were determined around 50 days after the incubation. In the Fukui paddy fields, DOC concentration varied seasonally from 1.1 to 10.1 mg.Cl(-1), showing higher concentration in heavy runoff of non agriculture period in April. However, DOC concentration variation did not always correspond to rainfall. The Obadake paddy fields also showed a similar pattern with Fukui paddy fields. The daily DOC discharge per area in Fukui (up), Fukui (down), Obadake (south), Obadake (north) paddy fields influent from paddy fields were 0.02, 0.0161, 0.0135 and 0.0027 kg.a(-1).day(-1), respectively. These differences resulted from differences in agricultural types and customs of farmers according to paddy fields and fields. Also, SUVA (an indirect means to evaluate humic substances (hydrophobic fraction)) of the studied influent waters from paddy fields were generally lower than the influent waters from crop fields. The non-biodegradable DOC accounted for 50.2-98%, 46.8-85.5% of the total DOC in the paddy fields and crop fields. PMID- 16477992 TI - Landfill leachate treatment by an experimental subsurface flow constructed wetland in tropical climate countries. AB - Municipal leachate was treated in an experimental unit of constructed wetlands of subsurface flow type. The parameters studied were organics (BOD and COD), solids and heavy metals (Zn, Ni, Cu, Cr and Pb). Using two types of emergent plants of Scirpus globulosus and Eriocaulon sexangulare, more than 80% removal was achieved for all the parameters. E. sexangulare removed organics and heavy metals better than Scirpus globulosus. A higher concentration of heavy metals in the influent did not change the removal efficiency. PMID- 16477993 TI - Natural recharge to sustainable yield from the barind aquifer: a tool in preparing effective management plan of groundwater resources. AB - This paper presents the results of water balance study and aquifer simulation modeling for preliminary estimation of the recharge rate and sustainable yield for the semi arid Barind Tract region of Bangladesh. The outcomes of the study are likely to be useful for planning purposes. It is found from detailed water balance study for the area that natural recharge rates in the Barind Tract vary widely year to year. It may have resulted from the method used for the calculation. If the considered time interval had been smaller than the monthly rainfall, the results could have been different. Aquifer Simulation Modeling (ASM) for the Barind aquifer is used to estimate long-term sustainable yield of the groundwater considering limiting drawdown from the standpoint of economic pumping cost. In managing a groundwater basin efficiently and effectively, evaluation of the maximum annual groundwater yield of the basin that can be withdrawn and used without producing any undesirable effect is one of the most important issues. In investigating such recharge rate, introduction of certain terms such as sustainable yield and safe yield has been accompanied. Development of this area involves proper utilization of this vast land, which is possible only through ensured irrigation for agriculture. The Government of Bangladesh has a plan to develop irrigation facilities by optimum utilization of available ground and surface water. It is believed that the groundwater table is lowering rapidly and the whole region is in an acute state of deforestation. Indiscriminate groundwater development may accelerate deforestation trend. In this context estimation of actual natural recharge rate to the aquifer and determination of sustainable yield will assist in proper management and planning of environmentally viable abstraction schemes. It is revealed from the study that the sustainable yield of ground water (204 mm/y) is somewhat higher than the long term annual average recharge (152.7 mm) to the groundwater reservoir. The reason behind this is that the rivers within and around the Barind Tract might have played the role of influent rivers. PMID- 16477994 TI - Development of a geographical information system for pesticide assessment on an Ecuadorian watershed. AB - Banana production is very important for the Ecuadorian national income, but it is also a potential source of river pollution in the country. Current research is focusing on the use of GIS to perform pesticide assessment of a river basin that includes banana as its main agricultural activity. The Agricultural Non Point Source Model (AGNPS) is being used to perform pesticide assessment in the basin. The model structure and type of needed data was identified. By applying GIS methodology, a complete set of cartographical information was obtained to supply input data to the pesticide model. In addition, ready-to-print maps were useful by-products for farmers. PMID- 16477995 TI - Flood impact on water quality of small urban streams. AB - Flood situation (August 2002) in small urban streams radically changed status of water ecosystem, which had been created naturally and anthropogenically for a long time. The aim of the after-flood research has been to record development of a creek recovery in positive as well as negative sense: recovery of benthic colonisation and reloading of contaminants (heavy metals) in bottom sediment. The Botic creek (its one section) was chosen as an experimental stream. This section of the creak is influenced by combine sewer system (two overflows - CSOs) as well as storm sewer system (one outlet - SSO). Heavy metals concentrations in water, sediment and body tissue of benthic organisms of the Botic creek and quality of benthic communities were monitored long term before flood. Comparison of results before and after flood can better assess impact of extreme water stage on a small stream. PMID- 16477996 TI - Development of on-site fish exposure system placed in water quality monitoring stations along a river. AB - Estrogen-like substances have been suspected to cause feminization of wild fish in rivers in Japan. To elucidate the influence of estrogen-like substances on fish in river, we have started to develop the on-site continuous fish exposure system using medaka Oryzias latipes that were placed in water quality monitoring stations along a river. Adult male medaka were exposed to the river water in a glass exposure tank placed in the monitoring stations. Flow rate of water and water temperature were controlled at 30 L/hour and 26 degrees C respectively, and a light: dark cycle was maintained 16:8 hours. A commercial diet free from phytoestrogens was fed 4 times in a day using automatic feeder. After 2-week exposure, hepatic vitellogenin concentration of each male medaka was measured. The exposure tests were repeatedly performed at both the upstream and the downstream of sewage treatment plants along the River Tama which is a representative urbanized river in Japan. At the control site Haijimabashi monitoring station, vitellogenin was not detected in male medaka. On the other hand, at the Ishihara monitoring station which is the most downstream in this test area, every male medaka were produced vitellogenin in the test performed in the spring of 2004. As the results of the water quality analysis, it could be inferred that the estrone derived from effluents of sewage treatment plants caused the feminization of male medaka. The reason why the concentrations of the estrone and the estrogenic activity using DNA recombinant yeast varied in proportion to the electric conductivity of river water measured at the water quality monitoring station. Furthermore, after continuous 2-week exposure, the vitellogenin production of male medaka was reduced similar to the decrease of the concentrations of the estrone and the estrogenic activity of river water. PMID- 16477997 TI - Sewage outfall plume dispersion observations with an autonomous underwater vehicle. AB - This work represents one of the first successful applications of Autonomous Underwater Vehicles (AUVs) for interdisciplinary coastal research. A monitoring mission to study the shape and estimate the initial dilution of the S. Jacinto sewage outfall plume using an AUV was performed on July 2002. An efficient sampling strategy enabling greater improvements in spatial and temporal range of detection demonstrated that the sewage effluent plume can be clearly traced using naturally occurring tracers in the wastewater. The outfall plume was found at the surface highly influenced by the weak stratification and low currents. Dilution varying with distance downstream was estimated from the plume rise over the outfall diffuser until a nearly constant value of 130:1, 60 m from the diffuser, indicating the near field end. Our results demonstrate that AUVs can provide high quality measurements of physical properties of effluent plumes in a very effective manner and valuable considerations about the initial mixing processes under real oceanic conditions can be further investigated. PMID- 16477998 TI - Testing of conductivity/calcium and rubidium/strontium ratios as indicators of the chemical stability of a river: comparison with a biological indicator. AB - It is customary to detect pollution in a water flow by monitoring the increase of sensitive elements concentrations (NH4+, PO4(3-), NO3-...). However, concentrations are dependent on the flow rate and these compounds are not conservative, implying a concentration decrease downstream leading to false negative diagnosis of pollution impact. The use of elemental ratios of conservative compounds should diminish these pitfalls. We then thought of the chi/Ca (conductivity/calcium) and Rb/Sr (rubidium/strontium) ratios as water chemical stability indicators to clearly identify and discriminate point from diffuse pollutions. This hypothesis has been tested on 12 brooks located in the basin of Lake Geneva, during 2 hydrological years. The results were compared to the observed land use of the watershed and a biological indicator: the Pollution Sensitivity Index (PSI). The PSI is calculated from diatom taxonomy and evaluates biological quality with a grade ranging from 0 to 20 (bad to excellent). The main results of the research can be summarized as follows. The pollution signal is observable far downstream of the pollution site. Both chi/Ca and Rb/Sr ratios are water quality indicators expressing the stability of water chemistry. They can both be used to detect diffuse and point pollution impact. These indicators provide complementary information: chi/Ca variations increase in case of point pollution; Rb/Sr variations increase when diffuse pollutions occur. The results obtained with the indicators chi/Ca and Rb/Sr agree with biological indicator and observation of the land use. chi/Ca and Rb/Sr ratios represent important tools to identify and discriminate point source pollution from diffuse pollution. PMID- 16477999 TI - In vitro interactions with the Cytochrome P450 system, toxicity, and local anaesthetic effects of Fomocaine Alkylmorpholine derivatives in rats. AB - Fomocaine (CAS 56583-43-8) is a basic ether-type local anaesthetic used in dermatological practice for surface anaesthesia. For many years, modifications of the fomocaine molecule have been pursued, e.g. to improve its affinity to the sodium channel and also in view of possible new (systemic) applications. In the present study fomocaine and eight fomocaine derivatives with an additional alkyl chain in 2- or 3-position of different length (C1 up to C4), or with a branched C3 chain in 3-position, respectively, at the morpholine ring were evaluated in vitro for possible structure-activity relationships with respect to the interactions with cytochrome P450 (CYP) mediated monooxygenase and oxidase functions using rat liver 9000 g supernatants or microsomes. Results were compared to in vivo data from rats on toxicity (LD50), paresis of the N. ischiadicus and surface and conduction anaesthesia (cornea, N. ischiadicus). In general, the influence of the derivatives on the CYP system was less than that of fomocaine, showing a further decline with enlarging chain length. Toxicity of the derivatives was comparable to that of fomocaine and lower only with the compound with a C4 alkyl chain in 2-position. The derivatives caused a stronger surface anaesthesia than fomocaine, exhibiting an additional increase with enlarging chain length. No clear-cut structure-activity relationships were observed with respect to paresis of the N. ischiadicus and to conduction anaesthesia. Especially the derivatives having a C2 or C4 chain in 2- or a C3 chain in 3 position, respectively, may be of interest for further investigations. In comparison to fomocaine they caused a stronger surface anaesthesia combined with a lower interaction capacity with the CYP system. PMID- 16478000 TI - Effects of amlodipine orotate on hypertension-related complications in spontaneously hypertensive rats. AB - Hypertension is a common problem in elderly patients, which usually requires chronic therapy under various physiological conditions including low gastric acidity (hypo- or anacidity). This study investigated a new salt type of amlodipine (CAS 88150-42-9) on blood pressure and hypertension-related complications in stroke-prone spontaneously hypertensive rats (SHR-SP). Amlodipine orotate was prepared by reacting orotic acid and amlodipine to increase the dissolution rate at higher gastric pH conditions. Twelve-week-old SHR-SP were randomly divided into five groups to receive either amlodipine orotate or amlodipine besylate (CAS 111470-99-6) at the doses of 3 and 10 mg/kg/day orally for four weeks. The age-matched normotensive Wistar Kyoto rats (WKY) served as the normal positive control group. The systolic blood pressure was reduced in the amlodipine treated SHR-SP in a dose-dependent manner with a similar potency irrespective of the salt type. Both amlodipines also reduced the left ventricular hypertrophy at high doses and concentration-dependently inhibited the Ca2+ induced contraction with a similar potency. Furthermore, semi quantitative analysis of a cerebral injury revealed that the two salts of amlodipine reduced the stroke-re-lated lesions to a similar degree. These results suggest that the amlodipine orotate is effective in terms of its effects on hypertension, cardiac hypertrophy and stroke-related cerebral damage in SHR-SP. PMID- 16478001 TI - Cardiovascular effects of KUR-1246, a new tetrahydronaphthalen derivative beta2 adrenoceptor agonist and a selective uterine relaxant. AB - The aim of this study was to assess the cardiovascular effects of KUR-1246 (CAS 194785-31-4, (-)-bis(2-{[(2S)-2-({(2R)-2-hydroxy-2-[4-hydroxy-3-(2-hydroxyethyl) phenyl] ethyl}amino)-1,2,3,4-tetrahydronaphthalen-7-yl]oxy}-N,N dimethylacetamide)monosulfate), a new beta2-adrenoceptor agonist tocolytic agent. In conscious dogs, the intravenous administration of KUR-1246 at 0.1 and 1 microg/kg had no effects on blood pressure, heart rate or femoral artery blood flow. KUR-1246 at 10 and 100 microg/kg significantly decreased blood pressure and increased heart rate. In the electrocardiograms, KUR-1246 did not affect QT intervals or QTc. In addition, the cardiac effects of KUR-1246 were evaluated in in vitro electrophysiological studies. KUR-1246 at 10 micromol/L did not affect action potential parameters (the maximal upstroke velocity, resting membrane potential, action potential amplitude and action potential durations) in isolated papillary muscles of guinea pigs or in the human ether-a-go-go related gene (HERG) tail current recorded from stably transfected human embryonic kidney (HEK) 293 cells. On the basis of these results, the effects of KUR-1246 in conscious dogs on the cardiovascular system appear to be limited to changes in blood pressure and heart rate. Therefore, KUR-1246 is unlikely to provoke ventricular arrhythmias by delaying the ventricular repolarization. PMID- 16478002 TI - Protein- and lipid modification of natural bovine surfactant. Effects in experimental lung failure with special consideration of the response in neonates. AB - Surfactant therapy does not lead to a uniform, optimal and sustained effect on gas exchange in neonatal respiratory distress syndrome (RDS) and acute respiratory distress syndrome (ARDS). The aim of the present study therefore was to compare the effects of a lipid-enriched and protein-modified natural surfactant preparations with a licensed, clinically used bovine surfactant preparation - SF-Ril (Alveofact). METHODS: SF-Ril was enriched with phosphatidylglycerol, sphingomyeline, phosphatidylethanolamine plasmalogen, phosphatidylethanolamine. Furthermore, SF-Ril was modified with increased surfactant protein B (SP-B)/surfactant protein C (SP-C) content and finally a mercaptoethanol (ME) treated preparation for breaking the sulfhydril bondage of SP-B/SP-C by chemical reduction in methylene chloride using ME was developed. Finally ME was removed by vacuum extraction. These modified surfactants were tested at a dosage of 100 mg/ kg each in a model of respiratory failure induced by lung lavage in male adult rats and compared with SF-Ril at an identical dosage. Mechanical ventilation was standardised with fraction of inspiratory oxygen (FiO2) 1.0 and time-cycled pressure limited ventilation 16/0.8 cmH2O before and 26/6 cmH2O (peak inspiratory pressure/positive endexpiratory pressure) after lung lavage (target arterial oxygen pressure [pa02] < 100 mmHg), respiratory rate 36/min, inspiration/expiration time ratio 1:2. RESULTS: During the observation period of 120 min, the sphingomyeline substituted and protein modified (ME reduced) surfactant preparations exerted improved and sustained oxygenation compared with SF-Ril. Similar effects were observed for tidal volumes. All other preparations were equal or inferior to SF-Ril in our model. CONCLUSION: For the development of surfactant preparations less prone for inactivation the above mentioned data may provide useful information, provided they can be confirmed in further investigations employing other alternative models. PMID- 16478003 TI - Pharmacokinetics of tenatoprazole, a newly synthesized proton pump inhibitor, in healthy male Caucasian volunteers. AB - The pharmacokinetics of tenatoprazole (CAS 113712-98-4), a newly synthesized proton pump inhibitor, and its metabolites TU-501 (sulfide form) and TU-502 (sulfone form) were investigated in an ascending-dose parallel-group study at the dose levels of 10, 20, 40, 80 and 120 mg. A total of 30 healthy Caucasian male volunteers (6 in each dose group) received a single dose at Day 1 (fasted state) and repeated doses from Day 14 to Day 20. CYP2C19 genotype status was determined in all subjects. Concentrations of tenatoprazole, TU-501 and TU-502 in plasma and urine were measured by a validated HPLC/MS/MS method. The single and multiple dose study provided reliable tolerance. After the single administrations, plasma concentrations reached a maximum between 2.5 and 4.3 h post dose, and thereafter decreased according to a terminal half live (T1/2) ranging from 4.8 to 7.7 h. Similar T1/2 were obtained on first and the last administration, and the steady state was reached after 5 days. Cmax and AUC increased linearly between 10 to 80 mg. However, with the 120 mg dose, the observed Cmax was higher than expected, especially at steady state. For TU-501 and TU-502 metabolites, Cmax and AUC increased linearly after repeated administration between 40 and 120 mg. PMID- 16478004 TI - Synthesis, antimicrobial, and anti-inflammatory activities of novel 2-[3-(1 adamantyl)-4-substituted-5-thioxo-1,2,4-triazolin-1-yl] acetic acids, 2-[3-(1 adamantyl)-4-substituted-5-thioxo-1,2,4-triazolin-1-yl]propionic acids and related derivatives. AB - The reaction of 3-(1-adamantyl)-4-substituted-1,2,4-triazoline-5-thiones 3a-g with sodium chloroacetate, in ethanolic sodium hydroxide yielded the corresponding N1-acetic acid derivatives 4a-g. The interaction of 3a-g with ethyl 2-bromopropionate in acetone, in the presence of potassium carbonate, yielded the corresponding N1-ethyl propionate derivatives 5a-g, which upon hydrolysis with aqueous sodium hydroxide afforded the corresponding propionic acid derivatives 6a g. Similarly, the reaction of 3-(1-adamantyl)-4-amino-1,2,4-triazoline-5-thione 7 with sodium chloroacetate in ethanolic sodium hydroxide yielded the corresponding N1-acetic acid derivative 8. On the other hand, the reaction of 2-(1-adamantyl) 1,3,4-oxadiazoline-5-thione 9 with sodium chloroacetate yielded the corresponding S-acetic acid derivative 10. Compounds 4a-g, 5b, 5c, 5g, 6a-g, 8 and 10 were tested for in vitro activities against a panel of Gram-positive and Gram-negative bacteria and the yeast-like pathogenic fungus Candida albicans. Several derivatives produced good or moderate activities particularly against Bacillus subtilis. In addition, the in vivo anti-inflammatory activities of these compounds were determined using the carrageenin-induced paw oedema method in rats. Compounds 4a, 4b, 4e, 4f, 6f, 6g and 10 produced good dose-dependent anti inflammatory activities. PMID- 16478006 TI - [Design and method of clinical trials of drugs used in the therapy of stable peripheral arterial occlusive disease]. AB - To prove the efficacy of the treatment of atherosclerotic stable peripheral arterial occlusive disease with drugs certain criteria are to be considered in the context of the design and performance of clinical trials. These criteria refer to the design of the study, randomisation, blinding, inclusion and exclusion criteria, definition of outcomes, methods of examination, sample size estimation, and evaluation of the results. If these criteria and conditions are not taken into consideration, false positive and negative results can arise. In the present study, the criteria to prove the efficacy of vasoactive substances in the treatment of atherosclerotic peripheral occlusive arterial diseases are presented. PMID- 16478005 TI - Bioequivalence study of two amoxicillin formulations. AB - A randomized single-dose crossover study was conducted in 24 healthy male volunteers to compare the bioavailability of two amoxicillin (CAS 26787-78-0) formulations, Glomox tablet (test) and a commercially available original preparation, amoxicillin capsule (reference). One thousand milligram of each formulation were administered after an overnight fast with a washout period of three days. Sixteen blood samples were collected over 10 h, amoxicillin concentrations in deproteinized serum were determined by a high performance liquid chromatographic (HPLC) assay, and pharmacokinetic parameters were analyzed by the standard non-compartmental method. Mean +/- SD maximum concentration (Cmax), time to reach maximum concentration (Tmax), area under the curve (AUC0- >t and AUC0-->infinity), and elimination half-life (t1/2) were 13.30 +/- 4.52 and 12.99 +/- 3.56 microg/ml, 1.92 +/- 0.76 and 2.02 +/- 0.62 h, 42.50 +/- 13.62 and 42.24 +/- 12.35 microg x h/ml, 46.31 +/- 13.23 and 46.08 +/- 12.14 microg x h/ml, and 1.54 +/- 0.39 and 1.48 +/- 0.48 h for the test and reference formulation, respectively. The parametric 90 % confidence intervals of the mean of the difference (test-reference) between log-transformed values of the two formulations were 92.61% to 109.50%, 92.83% to 109.12%, and 93.11% to 109.41% for AUC0-->t, AUC0-->infinity, and Cmax, respectively. The results indicate that the two formulations can be considered equivalent with regard to the rate and extent of absorption under fasting conditions. PMID- 16478007 TI - Helping people to feel better about themselves and their environment is an essential aspect of health promotion. PMID- 16478008 TI - Society meeting in Scotland. PMID- 16478009 TI - Aunty state. PMID- 16478010 TI - World-wide rises in obesity: minimal hopes of control. PMID- 16478011 TI - Improving the mental and emotional well-being of looked after children: connecting research, policy and practice. AB - The relationships between research, policy and practice in improving the mental and emotional well-being of looked after children are complex. There is a growing body of research that demonstrates that children in state care (or 'looked after children') are particularly vulnerable in terms of poor health and mental health outcomes. The current health and social care policy context is in the midst of tremendous change. Within this 'modernisation agenda' the mental health of looked after children has received welcome attention, and this in turn has stimulated a major expansion in specialist mental health services for looked after children across the UK. However, despite what we know about some of the problems looked after children face, we know very little about what is effective. Although the available evidence from research now gives a stronger indication about prevalence and identifiable risk factors for morbidity (or co-morbidity), it does little to assist in identifying what interventions are effective in improving mental health with this client group. Efforts now need to be made by practitioners, policy makers and researchers to collectively concentrate on filling some of the gaps in our knowledge. This includes making better use of the evaluations of current services to inform the development of these services. PMID- 16478012 TI - Periodontitis and pregnancy. AB - There is an increasing amount of evidence suggesting a systemic link between periodontal disease in the pregnant mother and pre-term low birth weight (PLBW). Severe periodontitis affects at least 10% of the general population. The aetiology of periodontitis is essentially a bacterially induced inflammatory reaction within the attachment surrounding the teeth. Maternal infection has been linked with pre-term delivery. Normal pregnancy itself is associated with inflammatory changes very similar to those found in sepsis. Because the infected periodontal tissues can act as a reservoir for both bacterial products and inflammatory cytokines, it may be possible that periodontal infection and the resultant inflammation could be linked with PLBW. Current understanding suggests that prostaglandins and proinflammatory cytokines play a pivotal role in the initiation process because of the close relationship of inflammation and infection. High levels of maternally or fetally derived cytokines such as tumour necrosis factor alpha (TNFalpha) may enhance amniochorionic and decidual interleukin six (IL-6) expression. Prostaglandin E2 (PGE2) has also been associated with periodontitis and PLBW. Periodontitis is a possible risk factor for PLBW with an odds ratio of 2.30. For the majority of individuals affected with periodontitis, the condition is symptom-free until the disease is more advanced. Therefore there is the need for medical carers of pregnant patients to increase the awareness among pregnant women themselves. Although there is plenty of evidence associating periodontitis with PLBW, interventional studies for the treatment of periodontitis measuring the impact on PLBW are few in number. Therefore more good quality clinical trials are required to address this issue. PMID- 16478013 TI - Caregiver perspectives in oral healthcare in an institutionalised elderly population without access to dental services: a pilot study. AB - AIM: The provision of dental services to elderly populations is a complicated area. When clients are cognitively impaired, a new set of variables are introduced as care can be influenced by designated family members, caregivers or administrators, who are responsible for the oral health care of the client and may influence the types of treatment the client will receive. Differences in attitudes and perceptions, with respect to oral health, may lead to better or worse access to care. The goals of this study are: (a) to gain a better understanding of the perceptions which may govern access and barriers to care within the institutionalised elderly population; and (b) to improve information on where and how to target educational and service resources, in order to reduce barriers to care. METHOD: A convenience (non-randomised) sample of 100 residents with caregivers was selected from nursing homes without organised access to oral healthcare services. A structured interview was conducted with 40 family members. Questions explored the importance and priorities of dental services for elderly people in institutions. RESULTS: Services which family members deemed of highest importance were dentures (both to replace teeth and adjustments on existing dentures), pain relief, emergency dentistry, fillings, cleanings, check-ups, tooth removal and treatment for advanced gum disease. Services that family members felt were less important were complex restorations, cleaning instruction to caregivers and seniors, and services by a specialist. Services considered to be of least importance were root canal treatment and implants. The top service priorities identified by the sample of family members included: cleanings to prevent mouth disease; check-ups including X-rays; fillings; dentures to replace teeth; and dental treatment to relieve pain. CONCLUSION: This study found that family members and caregivers would like a basic dental service including check ups and preventive care, with restorative, denture and surgical intervention when required. PMID- 16478014 TI - The assessment and implementation of mouth care in palliative care: a review. AB - AIM: From a review of the literature it is evident that the importance and maintenance of oral health for patients with cancer is recognised as an integral part of basic nursing care and yet from practical observation in the hospital environment together with reviewing the published literature it is clear that there appears to be limited evidence of regular assessment of the mouth or implementation of oral protocols on the ward. Furthermore there is a lack of adequate training in the assessment and implementation of mouth care by nurses during their general nurse training. The aim of the present study was to review existing literature published between 1995 and 1999 to determine whether mouth care was effectively assessed and implemented in the palliative care setting. METHOD AND RESULTS: Relevant articles selected from the 1995-1999 period highlighted the lack of published research in this very important aspect of care. Results from these published articles raised concern about the lack of training and education among nurses in the assessment of mouth care as well as the need to rationalise the assessment tools used on the ward or hospice. Furthermore, while it was recognised in palliative care that a multiprofessional approach was beneficial, in practice this did not appear to apply to mouth care with a few notable exceptions. CONCLUSION: This review has highlighted a number of inconsistencies in both the knowledge of mouth care and its implementation by nursing staff. The importance of establishing protocols and setting standards of care was also indicated in this review. It is clear that without effective assessment of the mouth, the appropriate implementation of care will not be delivered. The implementation of mouth care should not be anecdotal in nature but based on research (evidence-based) and this in turn will enable nurses to embrace an evidence-based practice (which will benefit their patients) that can be effectively audited. PMID- 16478015 TI - A survey of the physical health screening of all in-patients in a residential rehabilitation population. AB - AIM: It has been noted for many years that people with severe and enduring mental illness frequently have poor physical health. Their physical health needs often go unrecognised by the mental health team. This study aims to investigate the physical health status of all in-patients in a residential rehabilitation setting. METHODS: All the clients in a rehabilitation centre in Doncaster, England, were included in the health screening survey. The physical health of patients was assessed following the guidelines from 'Physical Health of the Severe and Enduring Mentally 111. A Training Pack for GP Educators'. RESULTS: Over one-third (35%) of the patients studied were found to be obese and more than one-half (53%) were heavy smokers. These compare with figures of 24% for obesity and 25% for smoking in the general population. CONCLUSIONS: The physical health needs of people residing in rehabilitation units should be a high priority for the mental health staff and regular surveys should be considered. If we are successful at encouraging the mental health team to be involved in annual physical health screening of patients with severe mental illness in the rehabilitation unit, we might improve their physical health and reduce mortality. PMID- 16478016 TI - Body mass index of Kuwaiti children aged 3-9 years: reference percentiles and curves. AB - AIM: The suitability of using the standards for body mass index (BMI), produced in the U.S. by the National Center for Health Statistics, for assessing overweight and obesity among children in Kuwait and other Arabian Gulf countries has not been examined. These standards were obtained from better-nourished and genetically different populations to those found in Kuwait and in other Gulf region countries. The purpose of this study was to develop BMI reference percentiles and curves appropriate for children aged 3-9 in these countries. METHOD: Attempts were made to include all healthy Kuwaiti kindergarten and elementary education children in this study The total sample was 113,013, comprising 55,053 males and 57,960 females. The children were measured for weight and height from which the BMI was calculated. Appropriate polynomial regression smoothing techniques were used to obtain the best-fitting percentile curves. RESULTS: At percentiles < or =25th, the BMI of boys exceeded that of girls. At the 50th percentile, boys' BMI was mostly higher than or equal to that of the girls except at age nine where it was lower At the 75th percentile, the BMI of both genders was similar, with exceptions at age six and nine years. At the 85th and 95th percentiles, girls' BMI was consistently higher than males. At the lowest percentile, the BMI of US children was higher than Kuwaiti, Saudi (starting at six) and Iranian children. The BMI of Kuwaiti children at higher percentiles was higher than that of Saudi, Iranian (except at age < four years) and US children. CONCLUSION: BMI curves for Kuwaiti children follow almost the same pattern as their US counterparts but with noticeable variations especially at the lower and higher percentiles. This study may reflect that western standards may not be directly applicable to assess the level of BMI in Kuwait and possibly in the neighbouring Gulf countries, since they may overestimate the levels of overweight, obesity and underweight. PMID- 16478018 TI - Polar satellite freeze. PMID- 16478017 TI - To banish a cancer. PMID- 16478019 TI - Bouncy proteins. PMID- 16478021 TI - Pinching out sulfur. PMID- 16478020 TI - Talking up enlightenment. PMID- 16478023 TI - It's dogged as does it. Retracing Darwin's footsteps in the Galapagos shatters a myth but reveals how revolutions in science actually evolve. PMID- 16478022 TI - Sizing up. PMID- 16478024 TI - Teach the science. PMID- 16478025 TI - Plasma accelerators. PMID- 16478026 TI - Intrigue at the immune synapse. PMID- 16478028 TI - Protecting New Orleans. PMID- 16478027 TI - Thwarting nuclear terrorism. PMID- 16478029 TI - Miniaturized power. PMID- 16478030 TI - Owning the stuff of life. PMID- 16478031 TI - Putting a face on the first president. PMID- 16478032 TI - Into the breach. PMID- 16478033 TI - My virtual war. PMID- 16478034 TI - What causes a fever? PMID- 16478035 TI - A great masquerader learns a new trick. PMID- 16478037 TI - Diagnosing and managing posttraumatic stress disorder. AB - In addition to being associated with combat, posttraumatic stress disorder (PTSD) also occurs in civilians exposed to severe trauma or serious illness. Manifestations of PTSD are varied and commonly include nonspecific physical symptoms, sleep disturbances, and psychological problems. Treatment with selective serotonin reuptake inhibitors (SSRIs) is usually effective. PMID- 16478036 TI - A hypertensive emergency in an obese young woman. PMID- 16478038 TI - Evaluating and managing hypogammaglobulinemia. AB - If a patient has frequent or recurrent bronchopulmonary or sinus infections, they may be due to low levels of immunoglobulins. This article describes common primary (idiopathic) and secondary forms of hypogammaglobulinemia and how to evaluate and manage them. PMID- 16478039 TI - Natriuretic peptide testing: a window into the diagnosis and prognosis of heart failure. AB - B-type natriuretic peptide (BNP) and N-terminal pro-BNP (NT-proBNP) levels can indicate a variety of heart problems, as well as general critical illness. BNP and NT-proBNP assays are useful for evaluating patients with acute dyspnea, as a low level of natriuretic peptide can help rule out congestive heart failure (CHF) and reduce reliance on echocardiography. Conversely, these assays can be particularly useful in recognizing CHF in a patient with acute dyspnea and a history of chronic obstructive pulmonary disease. However, clinical judgment must always be part of the evaluation of BNP or NT-proBNP assay results. PMID- 16478040 TI - Improved outcomes in nephrotic syndrome. AB - Nephrotic syndrome can now be treated effectively in most cases. All patients should be treated with a low-salt diet, diuretics to reduce edema, and statins to normalize serum lipid concentrations. Patients with nephrotic syndrome are prone to deep vein thrombophlebitis, renal vein thrombosis, and pulmonary emboli. Depending on the condition, additional treatment may include corticosteroids, angiotensin-converting enzyme (ACE) inhibitors, angiotensin II receptor blockers (ARBs), cyclosporine, cytotoxic agents, or mycophenolate. PMID- 16478041 TI - The clinical picture. An expanding skin lesion in a patient on immunosuppressive therapy. PMID- 16478042 TI - Multiple sclerosis: treating symptoms, and other general medical issues. AB - Multiple sclerosis (MS) has protean manifestations, and the care of people with MS presents many unique challenges. Clinicians can have an important impact on health, quality of life, and daily functioning by participating in open dialogue, tailoring focused treatment plans, and anticipating general medical needs. PMID- 16478043 TI - Clostridium difficile-associated disease: new challenges from an established pathogen. AB - Clostridium difficile-associated disease (CDAD) can range from uncomplicated diarrhea to sepsis and even death. CDAD rates and severity are increasing, possibly due to a new strain. Transmission of C difficile occurs primarily in health care facilities via the fecal-oral route following transient contamination of the hands of health care workers and patients; contamination of the patient care environment also plays an important role. PMID- 16478045 TI - Membrane release and destabilization of Arabidopsis RIN4 following cleavage by Pseudomonas syringae AvrRpt2. AB - The Arabidopsis RIN4 protein mediates interaction between the Pseudomonas syringae type III effector proteins AvrB, AvrRpm1, and AvrRpt2 and the Arabidopsis disease-resistance proteins RPM1 and RPS2. Confocal laser-scanning fluorescence microscopy following particle bombardment of tobacco leaf epidermal cells was used to examine the subcellular localization of fusions between GFP and RIN4 or several of its homologs and to examine the effects of cobombardment with AvrRpt2 or AvrRpml. This study showed that RIN4 was attached to the plasma membrane at its carboxyl terminus and that a carboxyl-terminal CCCFxFxxx prenylation or acylation (typically palmitoylation) motif, or both, was essential for this attachment. RIN4 was cleaved by AvrRpt2 at two PxFGxW motifs, one releasing a large portion of RIN4 from the plasma membrane and both exposing amino-terminal residues that destabilized the carboxyl-terminal cleavage products by targeting them for N-end ubiquitylation and proteasomal degradation. Plasma membrane localization of RIN4 was not affected by AvrRpml. RIN4 was found to be part of a protein family comprising two full-length homologs and at least 11 short carboxyl-terminal homologs. Representatives of this family, comprising a full-length RIN4 homolog and two short carboxyl-terminal RIN4 homologs, were also attached to the plasma membrane and cleaved near their amino termini by AvrRpt2, but in contrast to RIN4, the major portions of these proteins remained on the plasma membrane. N-end degradation may play a minor role in RIN4 degradation but probably plays a major role in the degradation of RIN4 homologs and is, therefore, a major pathogenic consequence of AvrRpt2 cleavage. PMID- 16478044 TI - Nematode-induced changes of transporter gene expression in Arabidopsis roots. AB - Root-knot plant-parasitic nematodes (Meloidogyne spp.) account for much of the damage inflicted to plants by nematodes. The feeding sites of these nematodes consist of "giant" cells, which have characteristics of transfer cells found in other parts of plants. Increased transport activity across the plasma membrane is a hallmark of transfer cells, and giant cells provide nutrition for nematodes; therefore, we initiated a study to identify the transport processes that contribute to the development and function of nematode-induced feeding sites. The study was conducted over a 4-week period, during which time the large changes in the development of giant cells were documented. The Arabidopsis ATH1 GeneChip was used to identify the many transporter genes that were regulated by nematode infestation. Expression of 50 transporter genes from 18 different gene families was significantly changed upon nematode infestation. Sixteen transporter genes were studied in more detail using real-time reverse-transcriptase polymerase chain reaction to determine transcript abundance in nematode-induced galls that contain giant cells and uninfested regions of the root. Certain genes were expressed primarily in galls whereas others were expressed primarily in the uninfested regions of the root, and a third group was expressed evenly throughout the root. Multiple transport processes are regulated and these may play important roles in nematode feeding-site establishment and maintenance. PMID- 16478046 TI - MtENOD11 gene activation during rhizobial infection and mycorrhizal arbuscule development requires a common AT-rich-containing regulatory sequence. AB - The MtENOD11 gene from the model legume Medicago truncatula is transcriptionally activated both in response to Sinorhizobium meliloti Nod factors and throughout infection of root tissues by the nitrogen-fixing microsymbiont. To identify the regulatory sequences involved in symbiosis-related MtENOD11 expression, a series of promoter deletions driving the beta-glucuronidase reporter gene were analyzed in transgenic M. truncatula roots. These studies have revealed that distinct regulatory regions are involved in infection-related MtENOD11 expression compared with preinfection (Nod factor-mediated) expression. In particular, the 257-bp promoter sequence immediately upstream from the start codon is sufficient for infection-related expression, but is unable to drive gene transcription in response to the Nod factor elicitor. This truncated promoter is also sufficient to confer MtENOD11 expression during both the arbuscular mycorrhizal (AM) association and the parasitic interaction with root-knot nematodes. Site-directed mutagenesis further showed that a previously identified nodule-specific AT-rich motif is required for high-level MtENOD11 expression during S. meliloti infection as well as during AM colonization. However, mutation of this motif does not affect gene expression associated with nematode-feeding sites. Taken together, these results suggest a close link between regulatory mechanisms controlling transcriptional early nodulin gene activation during both rhizobial and AM root endosymbioses. PMID- 16478047 TI - In planta secretion of a calreticulin by migratory and sedentary stages of root knot nematode. AB - Esophageal secretions from endoparasitic sedentary nematodes are thought to play key roles throughout plant parasitism, in particular during the invasion of the root tissue and the initiation and maintenance of the nematode feeding site (NFS) essential for nematode development. The secretion in planta of esophageal cell wall-degrading enzymes by migratory juveniles has been shown, suggesting a role for these enzymes in the invasion phase. Nevertheless, the secretion of an esophageal gland protein into the NFS by nematode sedentary stages has never been demonstrated. The calreticulin Mi-CRT is a protein synthesized in the esophageal glands of the root-knot nematode Meloidogyne incognita. After three-dimensional modeling of the Mi-CRT protein, a surface peptide was selected to raise specific antibodies. In planta immunolocalization showed that Mi-CRT is secreted by migratory and sedentary stage nematodes, suggesting a role for Mi-CRT throughout parasitism. During the maintenance of the NFS, the secreted Mi-CRT was localized outside the nematode at the tip of the stylet. In addition, Mi-CRT accumulation was observed along the cell wall of the giant cells that compose the feeding site, providing evidence for a nematode esophageal protein secretion into the NFS. PMID- 16478048 TI - GmCOI1, a soybean F-box protein gene, shows ability to mediate jasmonate regulated plant defense and fertility in Arabidopsis. AB - The F-box protein gene COI1 from Arabidopsis plays a fundamental role in response to jasmonates, which regulate plant root growth, pollen fertility, wounding and healing, and defense against pathogens and insects. Null mutations in COI1 were previously found to abolish all the jasmonate responses, and the Arabidopsis coil 1 mutant is male sterile and susceptible to pathogen infection. In this study, we isolated an F-box protein gene from soybean, which shares significant homology with the Arabidopsis COI1 and similarly contains an F-box motif and leucine rich repeats (LRR), here designated GmCOI1 (Glycine max L. (Merr.) COI1). To test whether the sequence homology and structural similarity are indicative of functional conservation, we expressed GmCOI1 in the Arabidopsis coil-1 mutant. The transgenic coil-1 plants with expression of the GmCOI1 gene were found to exhibit normal jasmonate responses, including jasmonate-regulated plant defense and fertility. In addition, the chimerical proteins with swapped domain of the F box motif or LRR between GmCOI1 and COI1 were shown to functionally complement the coil-1 mutation. Furthermore, GmCOI1 was found to assemble into the Skpl Cullin-F-box (SCF) complexes, similar to the formation of the Arabidopsis SCF(COO1). These data demonstrate the soybean F-box protein gene GmCOI1 is able to mediate jasmonate-regulated plant defense and fertility in Arabidopsis, which implies a generic jasmonate pathway with conserved signal components in different plant species. PMID- 16478049 TI - Pyramiding unmarked deletions in Ralstonia solanacearum shows that secreted proteins in addition to plant cell-wall-degrading enzymes contribute to virulence. AB - Ralstonia solanacearum, like many phytopathogenic bacteria, makes multiple extracellular plant cell-wall-degrading enzymes (CWDE), some of which contribute to its ability to cause wilt disease. CWDE and many other proteins are secreted to the milieu via the highly conserved type II protein secretion system (T2SS). R. solanacearum with a defective T2SS is weakly virulent, but it is not known whether this is due to absence of all the CWDE or the loss of other secreted proteins that contribute to disease. These alternatives were investigated by creating mutants of wild-type strain GMI1000 lacking either the T2SS or up to six CWDE and comparing them for virulence on tomato plants. To create unmarked deletions, genomic regions flanking the target gene were polymerase chain reaction (PCR)-amplified, were fused using splice overlap extension PCR, were cloned into a suicide plasmid harboring the sacB counter-selectable marker, and then, were site-specifically introduced into the genome. Various combinations of five deletions (delta pehA, delta pehB, delta B, PehC, and Pme) was not statistically different from GMI1000, but all the mutants lacking one or both cellulolytic enzymes (Egl or CbhA) wilted plants significantly more slowly than did the wild type. The GMI-6 mutant that lacks all six CWDE was more virulent than the mutant lacking only its two cellulolytic enzymes, and both were significantly more virulent than the T2SS mutant (GMI-D). Very similar results were observed in wounded-petiole inoculation assays, so GMI-6 and GMI-D appear to be less capable of colonizing tomato tissues after invasion. Because the T2SS mutant was much less virulent than the sixfold CWDE mutant, we conclude that other secreted proteins contribute substantially to the ability of R. solanacearum GMI1000 to systemically colonize tomato plants. PMID- 16478050 TI - Mutagenesis of all eight avr genes in Xanthomonas campestris pv. campestris had no detected effect on pathogenicity, but one avr gene affected race specificity. AB - Suppression subtractive hybridization (SSH) was used to identify genes present in the systemic crucifer black rot pathogen Xanthomonas campestris pv. campestris 528T but missing from the nonsystemic crucifer leaf spot pathogen, X. campestris pv. armoraciae 417. Among the DNA fragments unique to 528T was Xcc2109, one of eight putative avr genes identified in the published 528T genome (NC_003902). Individual and sequential deletion, insertion mutations, or both of all eight 528T avr gene loci were made, but no change in pathogenicity was observed with any combination of avr mutations, including a strain with all eight avr genes deleted. However, insertion or deletion mutants affecting the Xcc2109 locus lost avirulence (i.e., became virulent) on Florida Mustard, an X. campestris pv. campestris race-determining, differential host. The Xcc2109 open reading frame as annotated was cloned and found to be nonfunctional. A longer gene, encompassing Xcc2109 and here designated avrXccFM, was cloned and found to complement the Xcc2109 mutants and to confer avirulence to two additional wild-type X. campestris pv. campestris strains, thereby changing their races. Resistance in Florida Mustard to 528T strains carrying avrXccFM occurred without a typical hypersensitive response (HR) on leaves, although a vascular HR was observed in seedlings. PMID- 16478051 TI - The ability to detoxify the mycotoxin deoxynivalenol colocalizes with a major quantitative trait locus for Fusarium head blight resistance in wheat. AB - We investigated the hypothesis that resistance to deoxynivalenol (DON) is a major resistance factor in the Fusarium head blight (FHB) resistance complex of wheat. Ninety-six double haploid lines from a cross between 'CM-82036' and 'Remus' were examined. The lines were tested for DON resistance after application of the toxin in the ear, and for resistances to initial infection and spread of FHB after artificial inoculation with Fusarium spp. Toxin application to flowering ears induced typical FHB symptoms. Quantitative trait locus (QTL) analyses detected one locus with a major effect on DON resistance (logarithm of odds = 53.1, R2 = 92.6). The DON resistance phenotype was closely associated with an important FHB resistance QTL, Qfhs.ndsu-3BS, which previously was identified as governing resistance to spread of symptoms in the ear. Resistance to the toxin was correlated with resistance to spread of FHB (r = 0.74, P < 0.001). In resistant wheat lines, the applied toxin was converted to DON-3-O-glucoside as the detoxification product. There was a close relation between the DON-3 glucoside/DON ratio and DON resistance in the toxin-treated ears (R2 = 0.84). We conclude that resistance to DON is important in the FHB resistance complex and hypothesize that Qfhs.ndsu-3BS either encodes a DON-glucosyl-transferase or regulates the expression of such an enzyme. PMID- 16478052 TI - The coexistence of symbiosis and pathogenicity-determining genes in Rhizobium rhizogenes strains enables them to induce nodules and tumors or hairy roots in plants. AB - Bacteria belonging to the family Rhizobiaceae may establish beneficial or harmful relationships with plants. The legume endosymbionts contain nod and nif genes responsible for nodule formation and nitrogen fixation, respectively, whereas the pathogenic strains carry vir genes responsible for the formation of tumors or hairy roots. The symbiotic and pathogenic strains currently belong to different species of the genus Rhizobium and, until now, no strains able to establish symbiosis with legumes and also to induce tumors or hairy roots in plants have been reported. Here, we report for the first time the occurrence of two rhizobial strains (163C and ATCC11325T) belonging to Rhizobium rhizogenes able to induce hairy roots or tumors in plants and also to nodulate Phaseolus vulgaris under natural environmental conditions. Symbiotic plasmids (pSym) containing nod and nif genes and pTi- or pRi-type plasmids containing vir genes were found in these strains. The nodD and nifH genes of the strains from this study are phylogenetically related to those of Sinorhizobium strains nodulating P. vulgaris. The virA and virB4 genes from strain 163C are phylogenetically related to those of R. tumefaciens C58, whereas the same genes from strain ATCC 11325T are related to those of hairy root-inducing strains. These findings may be of high relevance for the better understanding of plant-microbe interactions and knowledge of rhizobial phylogenetic history. PMID- 16478053 TI - Amylopectin induces fumonisin B1 production by Fusarium verticillioides during colonization of maize kernels. AB - Fusarium verticillioides, a fungal pathogen of maize, produces fumonisin mycotoxins that adversely affect human and animal health. Basic questions remain unanswered regarding the interactions between the host plant and the fungus that lead to the accumulation of fumonisins in maize kernels. In this study, we evaluated the role of kernel endosperm composition in regulating fumonisin B1 (FB1) biosynthesis. We found that kernels lacking starch due to physiological immaturity did not accumulate FB1. Quantitative polymerase chain reaction analysis indicated that kernel development also affected the expression of fungal genes involved in FB1 biosynthesis, starch metabolism, and nitrogen regulation. A mutant strain of F. verticillioides with a disrupted a-amylase gene was impaired in its ability to produce FB1 on starchy kernels, and both the wild-type and mutant strains produced significantly less FB1 on a high-amylose kernel mutant of maize. When grown on a defined medium with amylose as the sole carbon source, the wild-type strain produced only trace amounts of FB1, but it produced large amounts of FB1 when grown on amylopectin or dextrin, a product of amylopectin hydrolysis. We conclude that amylopectin induces FB1 production in F. verticillioides. This study provides new insight regarding the interaction between the fungus and maize kernel during pathogenesis and highlights important areas that need further study. PMID- 16478054 TI - Functional genomic analysis of global regulator NolR in Sinorhizobium meliloti. AB - NolR is a regulator of nodulation genes present in species belonging to the genera Rhizobium and Sinorhizobium. The expression of the nolR gene in Sinorhizobium meliloti AK631 was investigated in relation to stage of growth, availability of nutrients, and different environmental stimuli using the nolR::lacZ fusion report system. It has been shown that the nolR gene is regulated in a population-density-dependent fashion and influenced by a number of environmental stimuli, including nutrients, pH, and oxygen. Exploration of the physiological functions of NolR under various laboratory conditions has shown that NolR is required for the optimal growth of the bacteria on solid media, optimal survival of the bacteria in carbon-starved minimal medium, and after heat shock challenge. NolR also is involved in recipient-induced conjugative transfer of a plasmid. Proteome analysis of strain AK631 and its Tn5-induced nolR deficient mutant EK698 revealed that a functional NolR induced significant differences in the accumulation of 20 polypeptides in peptide mass fingerprinting early-log-phase cultures and 48 polypeptides in stationary-phase cultures. NolR acted mainly as a repressor in the early-log-phase cultures, whereas it acted as both repressor and activator in the stationary-phase cultures. The NolR protein and 59 NolR-associated proteins have been identified by peptide mass fingerprinting. The NolR protein was differentially expressed only in the NolR+ wild-type strain AK631 but not in its NolR- derivative EK698, confirming that no functional NolR was produced in the mutant. The NolR-associated proteins have diverse functions in amino acid metabolism, carbohydrate metabolism, lipid metabolism, nucleotide metabolism, energy metabolism, metabolism of Co-factors, and cellular adaptation and transportation. These results further support our previous proposal that the NolR is a global regulatory protein which is required for the optimization of nodulation, bacterial growth and survival, and conjugative transfer of a plasmid. PMID- 16478055 TI - Accommodation with higher-order monochromatic aberrations corrected with adaptive optics. AB - Higher-order monochromatic aberrations in the human eye cause a difference in the appearance of stimuli at distances nearer and farther from best focus that could serve as a signed error signal for accommodation. We explored whether higher order monochromatic aberrations affect the accommodative response to 0.5 D step changes in vergence in experiments in which these aberrations were either present as they normally are or removed with adaptive optics. Of six subjects, one could not accommodate at all for steps in either condition. One subject clearly required higher-order aberrations to accommodate at all. The remaining four subjects could accommodate in the correct direction even when higher-order aberrations were removed. No subjects improved their accommodation when higher order aberrations were corrected, indicating that the corresponding decrease in the depth of field of the eye did not improve the accommodative response. These results are consistent with previous findings of large individual differences in the ability to accommodate in impoverished conditions. These results suggest that at least some subjects can use monochromatic higher-order aberrations to guide accommodation. They also show that some subjects can accommodate correctly when higher-order monochromatic aberrations as well as established cues to accommodation are greatly reduced. PMID- 16478056 TI - Analysis of dense-medium light scattering with applications to corneal tissue: experiments and Monte Carlo simulations. AB - Dense-medium scattering is explored in the context of providing a quantitative measurement of turbidity, with specific application to corneal haze. A multiple wavelength scattering technique is proposed to make use of two-color scattering response ratios, thereby providing a means for data normalization. A combination of measurements and simulations are reported to assess this technique, including light-scattering experiments for a range of polystyrene suspensions. Monte Carlo (MC) simulations were performed using a multiple-scattering algorithm based on full Mie scattering theory. The simulations were in excellent agreement with the polystyrene suspension experiments, thereby validating the MC model. The MC model was then used to simulate multiwavelength scattering in a corneal tissue model. Overall, the proposed multiwavelength scattering technique appears to be a feasible approach to quantify dense-medium scattering such as the manifestation of corneal haze, although more complex modeling of keratocyte scattering, and animal studies, are necessary. PMID- 16478057 TI - Gloss as an aspect of the measurement of appearance. AB - The perception and measurement of gloss is just one part of the concept of appearance measurement. We describe the comparison between data derived by 20 observers who each scaled the perceived gloss of a set of 84 neutral and colored test samples and measurements of those samples made using a glossmeter and a sphere-based spectrophotometer. With a glossmeter, the relationship between the visual-scaled data and the measured gloss values could be described by a three part linear fit or, with a higher correlation, a cubic function. For the sphere based spectrophotometer, the difference in the luminance factor, deltaY, between specular-included and specular-excluded measurements, was found to give a better linear correlation with the observer-scaled data. The color of the samples did not appear to significantly change the perceived gloss of the surface. PMID- 16478058 TI - Effects of incident illumination on in-line phase-contrast imaging. AB - Effects of incident illumination on phase-contrast images obtained by means of free-space propagation are investigated under the "transport-of-intensity" approximation. Analytical expressions for image intensity distribution are derived in the cases of coherent quasi-plane and quasi-spherical incident waves, as well as for spatially incoherent and quasi-homogeneous sources and some other types of sources. Practical methods for measuring the relevant parameters of the incident radiation are discussed together with formulas allowing one to calculate the effect of these parameters on the image intensity distribution. The results are expected to be useful in quantitative in-line imaging, phase retrieval, and tomography with polychromatic and spatially partially coherent radiation. As an application we present a method for simultaneous "automatic" phase retrieval and spatial deconvolution in in-line imaging of homogeneous objects using extended polychromatic x-ray sources. PMID- 16478059 TI - Elliptic Laguerre-Gaussian beams. AB - An analytical expression for the diffraction of an elliptic Laguerre-Gaussian (LG) beam is derived and analyzed. We show that a beam with even singularity order has nonzero axial intensity for any degree of ellipticity and at any finite distance z from the initial plane, whereas at z = 0 and z = infinity the axial intensity is zero. We show that for a beam with a small degree of ellipticity and even order of singularity, two isolated intensity zeroes appear in the Fresnel zone on a straight line at an angle of 45 deg or -45 deg, depending whether the beam's spin is right or left. The theoretical conclusions are confirmed by numerical simulation and physical experiments. PMID- 16478060 TI - Reconstruction of long-period fiber gratings from their core-to-core transmission function. AB - In order to reconstruct the structure of a long-period grating, both the complex core-to-core transmission function and the complex core-to-cladding transmission function should be known. However, in practice, only the core-to-core transmission function of the grating can be measured. We demonstrate theoretically the reconstruction of long-period gratings from only the core-to core transmission function. The reconstruction is performed by extracting the complex core-to-cladding transmission function of the grating from its core-to core transmission function. Generally, the extraction is not unique; however, we show that by writing an additional grating in cascade to the interrogated grating, a unique reconstruction can be obtained. In weak long-period gratings, only the amplitude of the core-to-core transmission function is needed to reconstruct the grating. The results of our work can enable the experimental reconstruction of long-period gratings from their transmission function as well as the development of novel distributed sensors. PMID- 16478061 TI - Study of the differential theory of lamellar gratings made of highly conducting materials. AB - Differential theory is said to be difficult to apply to surface-relief gratings made of metals with very high conductivity even though the formulation follows Li's Fourier factorization rules. Recently, Popov et al. [J. Opt. Soc. Am. 21, 199 (2004)] pointed out this difficulty and explained that its origin is related to the inversion of Toeplitz matrices constructed by the permittivity distribution inside the groove region. The current paper provides information about the differential theory for highly conducting gratings and considers the numerical instability problems. A stable calculation for lossless gratings is described, based on the extrapolation technique with the assumption of small losses. PMID- 16478062 TI - Statistics of specular points at a randomly rough surface. AB - Statistical parameters are obtained for an ensemble of specular points at a randomly rough Gaussian statistically isotropic surface at normal incidence. The joint probability density functions (PDFs) of specular point heights and total curvatures are derived separately for maxima, minima, and saddle points. The joint PDFs of brightness and surface elevations of specular points of different types are obtained analytically in an explicit form. PMID- 16478063 TI - Electromagnetic spatial coherence wavelets. AB - The recently introduced concept of spatial coherence wavelets is generalized to describe the propagation of electromagnetic fields in the free space. For this aim, the spatial coherence wavelet tensor is introduced as an elementary amount, in terms of which the formerly known quantities for this domain can be expressed. It allows for the analysis of the relationship between the spatial coherence properties and the polarization state of the electromagnetic wave. This approach is completely consistent with the recently introduced unified theory of coherence and polarization for random electromagnetic beams, but it provides further insight about the causal relationship between the polarization states at different planes along the propagation path. PMID- 16478064 TI - Modified two-flux approximation for identification of radiative properties of absorbing and scattering media from directional-hemispherical measurements. AB - A modified two-flux approximation is suggested for calculating the hemispherical transmittance and reflectance of a refracting, absorbing, and scattering medium in the case of collimated irradiation of the sample along the normal to the interface. The Fresnel reflection is taken into account in this approach. It is shown that the new approximation is rather accurate for the model transport scattering function. For an arbitrary scattering medium, the error of the modified two-flux approximation is estimated by comparison with the exact numerical calculations for the Henyey-Greenstein scattering function in a wide range of albedos and optical thicknesses. Possible applications of the derived analytical solution to identification problems are discussed. PMID- 16478065 TI - Reflectance optimization of inhomogeneous coatings with continuous variation of the complex refractive index. AB - A model is derived for the reflectance optimization of an inhomogeneous coating made of absorbing materials. The model is applicable mainly for spectral regions where no transparent materials are available, such as in the extreme ultraviolet. The complex refractive index is assumed to take values within a given continuous domain and in a given sequence. The coating design is generated through a series of layer elements with a small refractive-index contrast across interfaces; the thickness of the element is calculated in terms of the refractive-index increment at the interface. The coating is optimized element by element starting from the substrate. When the refractive index varies both continuously and smoothly, the thickness element is of first order in the refractive-index increment. Suggestions are given on how to optimize a more general coating that alternates continuous and smooth refractive-index domains along with discrete indices, which results in a succession of inhomogeneous coatings and finite layers. An example is given to illustrate the model. A new material selection rule is obtained to discriminate whether the addition of a material on top of a partly grown coating will increase or decrease the reflectance of the coating. As a consequence, the model, which is highlighted toward the maximization of reflectance, can be used analogously for reflectance minimization such as for anti-reflection coatings. PMID- 16478066 TI - Simulation of surface plasmon resonance of metallic nanoparticles by the boundary element method. AB - A set of new surface integral equations (Fredholm equations of the second kind) has been systematically derived from the Stratton-Chu formulation of Maxwell's equations for a two-dimensional TM mode to investigate the interactions of an incident electromagnetic wave with nanostructures, especially metals. With these equations, the surface components (the tangential magnetic field, the normal displacement, and the tangential electric field) on the boundary are solved simultaneously by the boundary-element method numerically. For nanometer-sized structures (e.g., dimension of 10 nm), our numerical results show that surface plasmon resonance causes a strong near-field enhancement of the electric field within a shallow region close to the interface of metal and dielectric. In addition, the corresponding pattern of the far-field scattering cross section is like a dipole. For the submicrometer-sized cases (dimension of several hundreds of nanometers), the numerical results indicate the existence of a standing wave on the backside surface of metals. This phenomenon could be caused by two surface plasmon waves that creep along the contour of metals clockwise and counterclockwise, respectively, and interfere with each other. PMID- 16478067 TI - New approach to determine the effects of polarization mode dispersion and chromatic dispersion on pulse and RF signals. AB - We present a novel analytical expression relating the output state of polarization (SOP) and the polarization mode dispersion (PMD) vector, including polarization-dependent chromatic dispersion (PCD), in terms of the angle of precession of the output SOP around the PMD vector. We derive a number of new expressions incorporating for the first time this angle of precession. First, a general relation to study the effect of differential group delay, PCD, and chromatic dispersion on pulses of arbitrary shapes is given. From this general relation, we derive expressions for pulse broadening and power penalty for Gaussian pulses. Moreover, a new expression for PMD-induced power fading for single-sideband modulated radio frequency signals is also derived. Measured experimental results are presented to support the derived expressions. PMID- 16478068 TI - Invariant polarimetric contrast parameters of light with Gaussian fluctuations in three dimensions. AB - We propose a rigorous definition of the minimal set of parameters that characterize the difference between two partially polarized states of light whose electric fields vary in three dimensions with Gaussian fluctuations. Although two such states are a priori defined by eighteen parameters, we demonstrate that the performance of processing tasks such as detection, localization, or segmentation of spatial or temporal polarization variations is uniquely determined by three scalar functions of these parameters. These functions define a "polarimetric contrast" that simplifies the analysis and the specification of processing techniques on polarimetric signals and images. This result can also be used to analyze the definition of the degree of polarization of a three-dimensional state of light with Gaussian fluctuations in comparison, with respect to its polarimetric contrast parameters, with a totally depolarized light. We show that these contrast parameters are a simple function of the degrees of polarization previously proposed by Barakat [Opt. Acta 30, 1171 (1983)] and Setala et al. [Phys. Rev. Lett. 88, 123902 (2002)]. Finally, we analyze the dimension of the set of contrast parameters in different particular situations. PMID- 16478069 TI - Single-scale spectroscopy of structurally colored butterflies: measurements of quantified reflectance and transmittance. AB - Butterfly scales generally have very elaborate structures in submicrometer size, and some of them show distinctive optical effects through interaction with light. We describe two methods to quantitatively characterize the optical properties of the individual scales in those structurally colored butterflies. Owing to the small dimensions of the scale and to the fact that the reflection and transmission are very diffuse, it is generally difficult to accurately measure the reflectance and transmittance. To overcome these difficulties, we have carefully constructed an optical system including an integrating sphere and investigated variously colored nine kinds of scale. It is shown that the obtained spectra clearly characterize the optical differences among those structurally colored scales and also the differences between structural and pigmentary colors. Further, we have performed the angle-resolved measurement of the reflected light to characterize the spatial pattern of reflection, which is closely related to the mechanism of reflection. PMID- 16478070 TI - Observations on "nonradiating surface sources": comment. AB - The proof, established in a recent paper [A. J. Devaney, "Nonradiating surface sources," J. Opt. Soc. Am. A 21, 2216 (2004)], of the existence of nonradiating surface sources formed by singlet-plus-doublet components whose generated fields vanish in either of the regions separated by a closed or infinite surface where the source resides is corroborated by means of an equivalent but slightly different formalism based on treatment of partial differential operators in a weak derivative or distributional sense. This approach yields a construction procedure applicable to a broad class of singular nonradiating sources. A fundamental question raised in that paper concerning the nonexistence of nontrivial nonradiating infinite planar sources that generate vanishing fields at both associated half-spaces is re-examined, with the conclusion that it is actually possible to mathematically construct such singular nonradiating sources as long as one allows for higher-order singularities such as certain combinations of singlet and triplet components. PMID- 16478071 TI - Influence of total reflection on the imaging quality of optical systems: comment. AB - A recent study of the influence of the Goos-Hanchen shift on the imaging properties of planar optical systems is commented on. It is shown that the model chosen for a description of the Goos-Hanchen shift must be modified considerably and that there is one more shift in total reflection that will also influence the imaging quality. PMID- 16478072 TI - Multilayer resonant subwavelength gratings: effects of waveguide modes and real groove profiles. AB - The boundary integral equation code PCGrate-S(X) is used to analyze diffraction on Hubble Space Telescope Cosmic Origins Spectrograph gratings at different boundary shapes and layer thicknesses. An effect of resonance anomalies excited in nonconformal dielectric layers overcoated on the surface of metallic grating on the efficiency is studied for the first time to our knowledge. Refractive indices (RIs) for bulk MgF2 taken from well-known references are found to be not suitable for thin optical layers at wavelengths between 115 and 170 nm. A method based on scale fitting of calculated and measured grating efficiencies is outlined for derivation of thin-film optical constants at hard to measure wavelengths. The calculated efficiency based on real boundary profiles and derived RIs of the G185M subwavelength grating is shown to fit within 9.6% or better to the measured data. PMID- 16478073 TI - Phase constraint for the waves diffracted by lossless symmetrical gratings at Littrow mount. AB - The energy conservation of grating diffraction is analyzed in a particular condition of incidence in which two incident waves reach a symmetrical grating from the two sides of the grating normal at the first-order Littrow mounting. In such a situation the incident waves generate an interference pattern with the same period as the grating. Thus in each direction of diffraction, interference occurs between two consecutive diffractive orders of the symmetrical incident waves. By applying only energy conservation and the geometrical symmetry of the grating profile to this problem it is possible to establish a general constraint for the phases and amplitudes of the diffracted orders of the same incident wave. Experimental and theoretical results are presented confirming the obtained relations. PMID- 16478074 TI - Asphere testing with a Fizeau interferometer based on a combined computer generated hologram. AB - Fizeau interferometers with an additional diffractive optical element are frequently used for measuring spherical and aspherical surfaces. We present a new (to our knowledge) optical test method, in which the Fizeau principle is now perfectly fulfilled by generating reference and measuring wavefront on the last optical surface, which carries a diffractive optical element. This method has been examined experimentally by testing a reference f/0.68 spherical mirror and can be applied identically for testing aspheres. Several advantages of this method are discussed and proved experimentally. PMID- 16478075 TI - Apodized multilevel diffractive lenses that produce desired diffraction-limited focal spots. AB - An apodized, multilevel diffractive lens that can produce a desired diffraction limited focal spot is proposed for many applications, such as focusing, imaging, optical storage, and optical trapping. The three key points for the design are the innovative idea of complex conjugate subzones, the use of Babinet's principle, and the equivalent-pupil (or aperture) function theory of diffractive focusing elements composed of concentric transparent rings. As a concrete example, we numerically design a mixed multilevel diffractive lens (the highest phase level is 8) to produce a diffraction-limited Gaussian focal spot. Some related problems, such as the validity range and the combination with high numerical-aperture refractive lenses, are also discussed. PMID- 16478076 TI - Designing Talbot array illuminators with phase-space optics. AB - The problem of designing Talbot array illuminators is revisited in the context of phase-space optics. It is shown that for Talbot array illuminators with optimum compression ratio the construction of phase-only grating profiles can be simplified significantly by using phase-space representations of optical signals. Based on the Wigner distribution function a graphical procedure is derived for obtaining the complete design of the array generator for a given compression ratio. The application of phase-space optics to other classes of Talbot array illuminators, and its use as part of numerical optimization algorithms, is considered as well. PMID- 16478077 TI - Analysis of the effects of bias phase and wavelength choice on the design of dual wavelength diffractive optical elements. AB - Diffractive optical elements (DOEs) are often used in pattern formation for display purposes. Constructing these images from two or more colors greatly enhances their visual effect. To achieve this with DOEs is not simple, as they are inherently wavelength specific. We discuss an algorithm for designing quantized elements that produce distinct intensity patterns in the far field for two wavelengths. The benefits of applying bias phase to the dual-wavelength problem are investigated. The difference between the best and the worst choice of bias phase is shown to produce a variation of up to 2% in the efficiency. The mean square error can vary by up to a factor of 2 between the best and the worst case. It is also critically important to understand how the values of the two wavelengths affect the result. We present an analysis of how choosing different pairs of wavelengths in the design process affects the quality of our results. PMID- 16478078 TI - Fitting-based determination of an effective medium of a metallic periodic structure and application to photonic crystals. AB - Effective permittivities of a metallic periodic structure for which the second order effective-medium theory does not yield correct results were obtained by numerically fitting to rigorous-coupled-wave analysis (RCWA). The calculated effective medium showed good agreement with RCWA and minimal deviation in the long-wavelength limit with variation in angle of incidence, grating depth, superstrate, and fill factor. In terms of the standard deviation, the effective medium was least affected by the change in grating depths. The calculated effective permittivities were applied to a three-dimensional metallic photonic crystal structure and produced a photonic bandgap that is consistent with published experimental data. PMID- 16478079 TI - Never underestimate the importance of asking "why?". PMID- 16478080 TI - Results of the 2005 AORN salary survey--trends for perioperative nursing. AB - AORN conducted its annual compensation survey for perioperative nurses in August 2005. A multiple regression model was used to examine how a variety of variables, including job title, education level, certification, experience, and geographic region, affect nursing compensation. This survey also examines the effect of other forms of compensation (eg, on-call compensation, overtime, bonuses, shift differential) on average base compensation rates. PMID- 16478081 TI - Arthroscopic hip surgery. AB - Many pathological entities that previously could be accessed only through large, open incisions now can be treated arthroscopically. Furthermore, arthroscopy has allowed certain conditions to be diagnosed that previously were unrecognized. Arthroscopic hip surgery has become an accepted surgical procedure with well defined indications and expected outcomes, primarily because of recent advances in surgical instrumentation and techniques. Hip arthroscopy is performed as a same-day procedure using a fracture table to apply distraction to the hip joint. Coordination among surgeons and perioperative nursing staff members is essential for achieving consistent results and minimizing complications during this technically demanding procedure. PMID- 16478082 TI - Future trends in minimally invasive surgery. AB - Health care costs in the United States are climbing annually at a staggering rate. Technology often is blamed for rising health care costs; however, some medical advances, such as minimally invasive surgery (MIS), are critical to improving health care efficiency, enhancing the quality of care provided, and decreasing overall expenses. Savings or profits relating to technology may be difficult to measure quantitatively, but when surgeons and perioperative personnel have reached the peak of the learning curve for performing MIS procedures, complication rates drop significantly. Patients are experiencing the benefits of MIS (eg, decreased postoperative pain, shorter hospital stays and often no need for an overnight stay, ability to return to work sooner), all of which encourage technologically savvy consumers to seek MIS options. PMID- 16478083 TI - Necrotizing fasciitis--the importance of early diagnosis and debridement. AB - Necrotizing fasciitis (NF) is a potentially life-threatening bacterial infection of the skin, deep subcutaneous tissue, and fascia. Early symptoms may be misdiagnosed as cellulitis. A hallmark symptom that distinguishes NF from cellulitis is severe local pain that is out of proportion to the size and type of the wound present. Early diagnosis and treatment of NF is imperative for a patient's survival. This article describes the pathophysiologic mechanisms, clinical manifestations, and treatment of NF, as well as implications for perioperative nursing. PMID- 16478084 TI - Improving specimen practices to reduce errors. PMID- 16478085 TI - The MEDiC Act of 2005: a new approach to safety. PMID- 16478086 TI - Second-order peak detection for multicomponent high-resolution LC/MS data. AB - The first step when analyzing multicomponent LC/MS data from complex samples such as biofluid metabolic profiles is to separate the data into information and noise via, for example, peak detection. Due to the complex nature of this type of data, with problems such as alternating backgrounds and differing peak shapes, this can be a very complex task. This paper presents and evaluates a two-dimensional peak detection algorithm based on raw vector-represented LC/MS data. The algorithm exploits the fact that in high-resolution centroid data chromatographic peaks emerge flanked with data voids in the corresponding mass axis. According to the proposed method, only 4 per thousand of the total amount of data from a urine sample is defined as chromatographic peaks; however, 94% of the raw data variance is captured within these peaks. Compared to bucketed data, results show that essentially the same features that an experienced analyst would define as peaks can automatically be extracted with a minimum of noise and background. The method is simple and requires a priori knowledge of only the minimum chromatographic peak width-a system-dependent parameter that is easily assessed. Additional meta parameters are estimated from the data themselves. The result is well-defined chromatographic peaks that are consistently arranged in a matrix at their corresponding m/z values. In the context of automated analysis, the method thus provides an alternative to the traditional approach of bucketing the data followed by denoising and/or one-dimensional peak detection. The software implementation of the proposed algorithm is available at http://www.anchem.su.se/peakd as compiled code for Matlab. PMID- 16478087 TI - Pressure-modulated differential scanning calorimetry. An approach to the continuous, simultaneous determination of heat capacities and expansion coefficients. AB - A new method is described that permits the continuous and synchronous determination of heat capacity and expansibility data. We refer to it as pressure modulated differential scanning calorimetry (PMDSC), as it involves a standard DSC temperature scan and superimposes on it a pressure modulation of preselected format. The power of the method is demonstrated using salt solutions for which the most accurate heat capacity and expansibility data exist in the literature. As the PMDSC measurements could reproduce the parameters with high accuracy and precision, we applied the method also to an aqueous suspension of multilamellar DSPC vesicles for which no expansibility data had been reported previously for the transition region. Excellent agreement was obtained between data from PMDSC and values from independent direct differential scanning densimetry measurements. The basic theoretical background of the method when using sawtooth-like pressure ramps is given under Supporting Information, and a complete statistical thermodynamic derivation of the general equations is presented in the accompanying paper. PMID- 16478088 TI - Pressure-modulated differential scanning calorimetry: theoretical background. AB - We demonstrate in this work and in the accompanying paper that it is possible to measure simultaneously heat capacity and expansibility of biomolecules in a single DSC experiment. In this study, we provide the theoretical basis for this new method based on rigorous statistical thermodynamics. The theoretical treatment presented here demonstrates that there are two additive contributions to the heat capacity at variable pressure, viz. (1) the isobaric heat capacity and (2) an expansibility term, and that these contributions can be experimentally separated to obtain simultaneously both heat capacity and expansibility in continuous DSC temperature scans preformed under pressure modulation. Equations that describe the mixed heat capacity and expansibility signal are derived, and experimental strategies as well as data extraction procedures are discussed. PMID- 16478089 TI - Construction of single chain variable fragment (ScFv) and BiscFv-alkaline phosphatase fusion protein for detection of Bacillus anthracis. AB - This paper describes an attempt for convenient and sensitive detection of Bacillus anthracis with single chain variable fragment (scFv)-based protein chip. Phage display technology was employed to generate scFv by using the protective antigen (PA) of B. anthracis for immunization. V(H) and V(L) genes of the scFv were amplified separately by reverse transcriptase-PCR from mRNA of immunized mice and then assembled into scFv gene with a linker DNA sequence. The scFv gene was inserted into a phagemid vector pCANTAB-5E and then transformed into Escherichia coli TG1 to yield recombinant phages after infection with helper phage M13KO7. After six rounds of panning with PA, the phage clones displaying scFv fragments of the antibody were selected by ELISA. One phage clone scFv-6w10 showing the strongest positive signal in ELISA was selected. To enhance the affinity of the scFv-6w10, a recombinant bivalent single-chain Fv antibody (biscFv-6w10) directed against PA was constructed and tested in functional assays. The affinity of the biscFv-6w10 was much higher than that of scFv-6w10 and reached 6.5 x 10(9) M(-1). An expression system was constructed for the production of E. coli alkaline phosphatase (EAP) labeled biscFv-6w10 (biscFv-6w10 EAP) in E. coli cells. The expressed fusion protein retained both antigen specific binding and enzymatic activity and thus directly served as an enzyme labeled antibody. Detections of PA and bacterial cells of B. anthracis using biscFv-6w10-EAP and Cy3-labeled biscFv-6w10 were performed on a protein chip. The fusion protein (biscFv-6w10-EAP) chip could detect 10 pg of PA and 500-1000 bacterial cells in approximately 2 h, while the sensitivity of Cy3-labeled protein chip reached 1 pg of PA and 50-100 cells within 2 h. PMID- 16478090 TI - Probing protein ligand interactions by automated hydrogen/deuterium exchange mass spectrometry. AB - Amide hydrogen/deuterium exchange is a powerful biophysical technique for probing changes in protein dynamics induced by ligand interaction. The inherent low throughput of the technology has limited its impact on drug screening and lead optimization. Automation increases the throughput of H/D exchange to make it compatible with drug discovery efforts. Here we describe the first fully automated H/D exchange system that provides highly reproducible H/D exchange kinetics from 130 ms to 24 h. Throughput is maximized by parallel sample processing, and the system can run H/D exchange assays in triplicate without user intervention. We demonstrate the utility of this system to differentiate structural perturbations in the ligand-binding domain (LBD) of the nuclear receptor PPARgamma induced upon binding a full agonist and a partial agonist. PPARgamma is the target of glitazones, drugs used for treatment of insulin resistance associated with type II diabetes. Recently it has been shown that partial agonists of PPARgamma have insulin sensitization properties while lacking several adverse effects associated with full agonist drugs. To further examine the mechanism of partial agonist activation of PPARgamma, we extended our studies to the analysis of ligand interactions with the heterodimeric complex of PPARgamma/RXRalpha LBDs. To facilitate analysis of H/D exchange of large protein complexes, we performed the experiment with a 14.5-T Fourier transform ion cyclotron resonance mass spectrometer capable of measuring mass with accuracy in the ppb range. PMID- 16478091 TI - Analysis of glycosyl bond cleavage and related isotope effects in collision induced dissociation quadrupole-time-of-flight mass spectrometry of isomeric trehaloses. AB - The configuration isomers alpha,alpha-, alpha,beta-, and beta,beta-trehalose are distinguishable by a relative ion abundance analysis using collision-induced dissociation MS/MS measurements in electrospray ionization quadrupole-time-of flight mass spectrometry. The relative abundance of the Y-type fragment ion of alpha,alpha-trehalose is the highest and that of beta,beta-trehalose is the lowest, indicating that alpha-glycosyl bonds cleave more easily than beta glycosyl bonds. The relative ion abundance depends on both the alpha- and beta glycosyl linkage type and the number of alpha-glycosyl bonds. The reaction path of glycosyl bond cleavage is calculated computationally using the molecular orbital method in the form of Hartree-Fock theory in conjunction with the 6 31G(d) basis set. The results are consistent with the experimental data. Isotope effects on the fragmentation of the glycosyl bonds are detected in the experiments of the H2O/D2O solvent systems. Furthermore, the isotope effect regarding beta,beta-trehalose is larger than those of alpha,alpha- and alpha,beta trehalose, indicating that the isotope effect on the beta-glycosyl bond cleavage is larger than that on the alpha-glycosyl bond cleavage. The thermal energy increase in trehalose-d8 molecules over the corresponding trehalose molecules is calculated from the vibrational modes. PMID- 16478092 TI - Fiber-optic microsphere-based arrays for multiplexed biological warfare agent detection. AB - We report a multiplexed high-density DNA array capable of rapid, sensitive, and reliable identification of potential biological warfare agents. An optical fiber bundle containing 6000 individual 3.1-mum-diameter fibers was chemically etched to yield microwells and used as the substrate for the array. Eighteen different 50-mer single-stranded DNA probes were covalently attached to 3.1-mum microspheres. Probe sequences were designed for Bacillus anthracis, Yersinia pestis, Francisella tularensis, Brucella melitensis, Clostridium botulinum, Vaccinia virus, and one biological warfare agent (BWA) simulant, Bacillus thuringiensis kurstaki. The microspheres were distributed into the microwells to form a randomized multiplexed high-density DNA array. A detection limit of 10 fM in a 50-microL sample volume was achieved within 30 min of hybridization for B. anthracis, Y. pestis, Vaccinia virus, and B. thuringiensis kurstaki. We used both specific responses of probes upon hybridization to complementary targets as well as response patterns of the multiplexed array to identify BWAs with high accuracy. We demonstrated the application of this multiplexed high-density DNA array for parallel identification of target BWAs in spiked sewage samples after PCR amplification. The array's miniaturized feature size, fabrication flexibility, reusability, and high reproducibility may enable this array platform to be integrated into a highly sensitive, specific, and reliable portable instrument for in situ BWA detection. PMID- 16478093 TI - Chemical gas sensor application of open-pore mesoporous thin films based on integrated optical polarimetric interferometry. AB - Chemical gas sensors that employ integrated optical polarimetric interferometry were fabricated by the sol-gel synthesis of transparent mesoporous thin films of TiO2-P2O5 nanocomposite on tapered layers of TiO2 sputtered on tin-diffused glass waveguides. Atomic force microscopy images of the mesoporous thin film clearly show the open pore mouths on the film surface that favor rapid diffusion and adsorption of gas-phase analytes within the entire film. Adsorption of gas and vapor induces changes (Deltan) in the refractive index of the mesoporous thin film that lead to shifts in the phase difference between the fundamental transverse electric and magnetic modes simultaneously excited in the glass waveguide via single-beam incidence. Upon exposure to NH3 gas at concentrations as low as 100 ppb in dry air at room temperature, the sensor exhibits a reversible change in the phase difference with the response and recovery times of less than 60 and 90 s, respectively. It is unexpected that the sensor is unresponsive to either NO2 or C6H6 vapor, leading to a somewhat selective sensitivity to NH3. Determination of Deltan was carried out with a combination of the experimental results and the theoretical calculations. The sensor design represents a novel, effective, and easily accessible approach to mesoporous thin film-based integrated optical chemical sensors. PMID- 16478094 TI - Design and characterization of poly(dimethylsiloxane)-based valves for interfacing continuous-flow sampling to microchip electrophoresis. AB - This work describes the fabrication and evaluation of a poly(dimethyl)siloxane (PDMS)-based device that enables the discrete injection of a sample plug from a continuous-flow stream into a microchannel for subsequent analysis by electrophoresis. Devices were fabricated by aligning valving and flow channel layers followed by plasma sealing the combined layers onto a glass plate that contained fittings for the introduction of liquid sample and nitrogen gas. The design incorporates a reduced-volume pneumatic valve that actuates (on the order of hundreds of milliseconds) to allow analyte from a continuously flowing sampling channel to be injected into a separation channel for electrophoresis. The injector design was optimized to include a pushback channel to flush away stagnant sample associated with the injector dead volume. The effect of the valve actuation time, the pushback voltage, and the sampling stream flow rate on the performance of the device was characterized. Using the optimized design and an injection frequency of 0.64 Hz showed that the injection process is reproducible (RSD of 1.77%, n = 15). Concentration change experiments using fluorescein as the analyte showed that the device could achieve a lag time as small as 14 s. Finally, to demonstrate the potential uses of this device, the microchip was coupled to a microdialysis probe to monitor a concentration change and sample a fluorescein dye mixture. PMID- 16478095 TI - Integrated description of electrode/electrolyte interfaces based on equivalent circuits and its verification using impedance measurements. AB - An integrated theory describing both faradaic and nonfaradaic currents obtained upon potential step at an electrified electrode/electrolyte interface has been developed based on equivalent circuits that had been used to explain electrochemical reactions and experimentally verified. The faradaic current is shown to consist of the mass transport-dependent and -independent parts, which is in general agreement with the expression previously derived from the diffusion equations. The decay of the capacitive current is determined by the time constant represented by the product of the resistance obtained from the parallel connection of the solution and polarization resistances and the double layer capacitance; this is not consistent with the current understanding of the capacitive current decay, which takes into account the double layer capacitance and the solution resistance only. Many insights into the electron-transfer reactions are discussed based on the interpretation of impedance representation of the system, which would not have been possible without the present theory. PMID- 16478096 TI - Counting single native biomolecules and intact viruses with color-coded nanoparticles. AB - Nanometer-sized particles such as semiconductor quantum dots and energy-transfer nanoparticles have novel optical properties such as tunable light emission, signal brightness, and multicolor excitation that are not available from traditional organic dyes and fluorescent proteins. Here we report the use of color-coded nanoparticles and dual-color fluorescence coincidence for real-time detection of single native biomolecules and viruses in a microfluidic channel. Using green and red nanoparticles to simultaneously recognize two binding sites on a single target, we demonstrate that individual molecules of genes, proteins, and intact viruses can be detected and identified in complex mixtures without target amplification or probe/target separation. Real-time coincidence analysis of single-photon events allows rapid detection of bound targets and efficient discrimination of excess unbound probes. Quantitative studies indicate that the counting results are remarkably precise when the total numbers of counted molecules are more than 10. The use of bioconjugated nanoparticle probes for single-molecule detection is expected to have important applications in ultrasensitive molecular diagnostics, bioterrorism agent detection, and real-time imaging and tracking of single-molecule processes inside living cells. PMID- 16478097 TI - Improving sensitivity in shotgun proteomics using a peptide-centric database with reduced complexity: protease cleavage and SCX elution rules from data mining of MS/MS spectra. AB - Correct identification of a peptide sequence from MS/MS data is still a challenging research problem, particularly in proteomic analyses of higher eukaryotes where protein databases are large. The scoring methods of search programs often generate cases where incorrect peptide sequences score higher than correct peptide sequences (referred to as distraction). Because smaller databases yield less distraction and better discrimination between correct and incorrect assignments, we developed a method for editing a peptide-centric database (PC-DB) to remove unlikely sequences and strategies for enabling search programs to utilize this peptide database. Rules for unlikely missed cleavage and nontryptic proteolysis products were identified by data mining 11 849 high-confidence peptide assignments. We also evaluated ion exchange chromatographic behavior as an editing criterion to generate subset databases. When used to search a well annotated test data set of MS/MS spectra, we found no loss of critical information using PC-DBs, validating the methods for generating and searching against the databases. On the other hand, improved confidence in peptide assignments was achieved for tryptic peptides, measured by changes in DeltaCN and RSP. Decreased distraction was also achieved, consistent with the 3-9-fold decrease in database size. Data mining identified a major class of common nonspecific proteolytic products corresponding to leucine aminopeptidase (LAP) cleavages. Large improvements in identifying LAP products were achieved using the PC-DB approach when compared with conventional searches against protein databases. These results demonstrate that peptide properties can be used to reduce database size, yielding improved accuracy and information capture due to reduced distraction, but with little loss of information compared to conventional protein database searches. PMID- 16478098 TI - Combined approach to the analysis of recombinant protein drugs using hollow-fiber flow field-flow fractionation, mass spectrometry, and chemiluminescence detection. AB - The impurities present in recombinant protein drugs produced by large-scale refolding processes can not only affect the product safety but also interact with the expressed protein. To relate the impurity profile to conformation and functionality of the protein drug, analytical methods able not to degrade the sample components should be preferred. In this work, an urate oxidase (uricase) drug from Aspergillus flavus expressed in Saccharomyces cerevisiae, and a reagent grade uricase from Candida sphaerica expressed in Escherichia coli, are analyzed by combining hollow-fiber flow field-flow fractionation with matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI/TOFMS) and with chemiluminescence enzyme activity assay. Preliminary detection and identification of sample impurities is performed by means of conventional methods such as RP HPLC with electrospray ionization quadrupole-TOF MS and MALDI/TOFMS with SDS PAGE and 2D SDS PAGE. Results show that the recombinant uricase samples obtained from different microorganisms have different impurities and different enzymatic activity and that different uricase oligomers are present in solution. PMID- 16478099 TI - Determination and characterization of site-specific N-glycosylation using MALDI Qq-TOF tandem mass spectrometry: case study with a plant protease. AB - MALDI tandem mass spectrometry analysis on a hybrid quadrupole-quadrupole time-of flight (Qq-TOF) instrument was used in combination with two-dimensional gel electrophoresis, proteolytic digestion, and liquid chromatography for identification and structural characterization of glycosylation in a novel glycoprotein, pathogenesis-related subtilisin-like proteinase P69B from tomato. Glycopeptide fractions from microcolumn reversed-phase HPLC deposited on MALDI targets were identified from MS by their specific m/z spacing patterns (203, 162, 146 u) between glycoforms. In most cases, MS/MS spectra of [M + H]+ ions of glycopeptides featured peaks useful for determining sugar compositions, peptide sequences, and thus probable glycosylation sites. Furthermore, peptide-related product ions could readily be used in database search procedures to identify the glycoprotein. Four out of five predicted glycosylation sites were biologically relevant and occupied by five N-linked glycan side chains each. In addition, the fragmentation efficiency allowed detection of further modification of methionine containing glycoforms with either oxidized or iodoacetamide alkylated methionine. The high resolution furnished by MALDI-Qq-TOF allowed rapid and sensitive structural characterization of site-specific N-glycosylation from a limited quantity of material and revealed heterogeneity at different levels, including different glycan side-chain modifications, and heterogeneity of oligosaccharide structures on the same glycosylation site. PMID- 16478100 TI - Fluoroimmunoassay for antigen based on fluorescence quenching signal of gold nanoparticles. AB - A unique, sensitive, and highly specific fluoroimmunoassay system for antigen detection using gold and magnetic nanoparticles has been developed. The assay is based on the fluorescence quenching of fluorescein isothiocyanate caused by gold nanoparticles coated with monoclonal antibody. To demonstrate its analytical capabilities, the magnetic nanoparticles were coated with anti-alpha-fetoprotein polyclonal antibodies, which specifically bound with alpha-fetoprotein. Gold nanoparticles coated with anti-alpha-fetoprotein monoclonal antibodies could sandwich the alpha-fetoprotein captured by the magnetic nanoparticle probes. The sandwich-type immunocomplex was formed on the surface of magnetic nanoparticles and could be separated by a magnetic field. The supernatant liquid, which contained the unbound gold nanoparticle probes, was used to quench the fluorescence, and the fluorescence intensity of fluorescein isothiocyanate at 516 nm was proportional to the alpha-fetoprotein concentration. The result showed that the limit of detection of alpha-fetoprotein was 0.17 nM. This new system can be extended to detect target molecules with matched antibodies and has broad potential applications in immunoassay and disease diagnosis. PMID- 16478101 TI - Biosensor recognition of thyroid-disrupting chemicals using transport proteins. AB - Novel surface plasmon resonance-based biosensor assays for the bioeffect-related screening of chemicals with thyroid-disrupting activity are described. Two thyroid transport proteins (TPs), thyroxine binding globulin (TBG) and recombinant transthyretin (rTTR), were applied in an inhibition assay format in a Biacore 3000 using CM5 biosensor chips coated with l-thyroxine (T4), the main hormone of the thyroid system. Assay conditions were optimized for the natural thyroid hormones, and known thyroid disruptors and structurally related compounds were selected as model compounds to be tested in both assays for their relative potency (RP) compared to T4. The chosen compounds were halogenated phenols, halogenated bisphenols, bisphenol A, 3,5-dichlorobiphenyl, and its hydroxylated metabolite 4-hydroxy-3,5-dichlorobiphenyl (4-OH PCB 14). The TBG-based assay was highly specific for T4, and the rTTR-based assay was sensitive toward several compounds, the highest sensitivity (RP = 4.4) being obtained with 4-OH PCB 14, followed by tetrabromobisphenol A (RP = 1.5) and tetrachlorobisphenol A (RP = 0.75). For the bioeffect-related screening of known and identification of possible new thyroid disruptors, the TPs-based biosensor assays were more sensitive (IC(50) of 13.7 +/- 1.3 and 8.6 +/- 0.7 nM for the rTTR and the TBG based assay, respectively), easier to perform, and faster alternatives (10 min/sample) than the currently used methods such as radioligand binding assays and immunoprecipitation-HPLC. PMID- 16478102 TI - Electrochemical immunosensor for cholera toxin using liposomes and poly(3,4 ethylenedioxythiophene)-coated carbon nanotubes. AB - A sensitive method for the detection of cholera toxin (CT) using an electrochemical immunosensor with liposomic magnification followed by adsorptive square-wave stripping voltammetry is described. Potassium ferrocyanide encapsulated and ganglioside (GM1)-functionalized liposomes act as highly specific recognition labels for the amplified detection of cholera toxin. The sensing interface consists of monoclonal antibody against the B subunit of CT that is linked to poly(3,4-ethylenedioxythiophene) coated on Nafion-supported multiwalled carbon nanotube caste film on a glassy carbon electrode. The CT is detected by a "sandwich-type" assay on the electronic transducers, where the toxin is first bound to the anti-CT antibody and then to the GM1-functionalized liposome. The potassium ferrocyanide molecules are released from the bounded liposomes on the electrode by lyses with methanolic solution of Triton X-100. The released electroactive marker is measured by adsorptive square-wave stripping voltammetry. The sandwich assay provides the amplification route for the detection of the CT present in ultratrace levels. The calibration curve for CT had a linear range of 10(-14)-10(-7)g mL(-1). The detection limit of this immunosensor was 10(-16) g of cholera toxin (equivalent to 100 microL of 10(-15) g mL(-1)). PMID- 16478103 TI - Small-volume detection of Plasmodium falciparum CSP gene using a 50-microm diameter cavity with self-contained electrochemistry. AB - An electrochemical enzyme-linked immobilized DNA-hybridization assay for the detection of Plasmodium falciparum has been developed. The target molecule was a segment of the repeat sequence of the gene coding for the circumsporozoite (CSP) protein from the AF54087 gene. This analyte offers the possibility of specifically detecting P. falciparum. The assay involves attachment of a biotinylated primary DNA probe via its 5'-amine-terminus to the streptavidin coated surface of microwells in a 96-well plate. The primary DNA probe (1(0)P, which was of two different sequences we call 1(0)P(a) and 1(0)P(b)) was used to capture the target (T, which was of two different sequences, T1 sequence 481-590 and T2 sequence 472-590 of AF54087 gene for the CSP gene) by hybridization to a complementary sequence on the target. On 1(0)P(a), 47 bases were complementary to T1 and T2 at 543-590, while on 1(0)P(b), 35 bases were complementary to T1 and T2 at 555-590. A secondary DNA probe that contained 36 bases with alkaline phosphatase (2(0)P-AP) label on the 3' end was hybridized to a complementary base sequence on the 5' end of the target. p-Aminophenol, which is enzymatically generated by the immobilized AP from p-aminophenyl phosphate (PAPP), is detected using electrochemistry. The peak current of cyclic voltammograms from a PAPP solution incubated inside the microwells modified with the complete assembly of the assay components gives a linear relationship with the concentration of the target (2-50 ng/mL, where P1 (P1a and P1b) and P2-AP concentrations are 50 ng/mL). A detection limit of 1.4 ng/mL (or 46 pM) of the DNA target was obtained. The signals of the assays were not significantly affected when performed in the presence of human hepatocytes, pig liver, or chicken serum indicating the viability of this assay in real clinical samples. PMID- 16478104 TI - Fabrication of porous polymer monoliths in polymeric microfluidic chips as an electrospray emitter for direct coupling to mass spectrometry. AB - Coupling of polymeric microfluidic devices to mass spectrometry is reported using porous polymer monoliths (PPM) as nanoelectrospray emitters. Lauryl acrylate-co ethylene dimethacrylate porous polymer monolith was photopatterned for 5 mm at the end of the channel of microfluidic devices fabricated from three different polymeric substrate materials, including the following: poly(dimethylsiloxane) (PDMS), poly(methyl methacrylate) (PMMA), and cyclic olefin copolymer (COC). Spraying directly from the end of the chip removes any dead volume associated with inserted emitters or transfer lines, and the presence of multiple pathways in the PPM prevents the clogging of the channels, which is a common problem in conventional nanospray emitters. Spraying from a microfluidic channel having a PPM emitter produced a substantial increase in TIC stability and increased sensitivity by as much as 70x compared to spraying from an open end chip with no PPM. The performance of PPM emitter in COC, PMMA, and PDMS chips was compared in terms of stability and reproducibility of the electrospray. COC chips showed the highest reproducibility in terms of chip-to-chip performance, which can be attributed to the ease and reproducibility of the PPM formation due to the favorable optical and chemical properties of COC. We have further tested the performance of the COC chips by constant infusion of poly(propylene glycol) solution at organic content ranging from 10 to 90% methanol and at flow rates ranging from 50 to 1000 nL/min, showing optimum spraying conditions (RSD < 5%) at 50-70% organic content and at flow rates from 100 to 500 nL/min. The PPM sprayer was also used for protein preconcentration and desalting prior to mass spectrometric detection, and results were comparable with a chip spraying from an electrospray tip. PMID- 16478105 TI - Label-free DNA detection based on modified conducting polypyrrole films at microelectrodes. AB - A label-free electrochemical detection method for DNA hybridization based on electrostatic modulation of the ion-exchange kinetics of a polypyrrole film deposited at microelectrodes is reported. Synthetic single-stranded 27-mer oligonucleotides (probe) have been immobilized at 2,5-bis(2-thienyl)-N-(3 phosphorylpropyl)pyrrole film formed by electropolymerization on the previously formed polypyrrole layer. The 27- or 18-mer target oligonucleotides were monitored via the electrochemically driven anion exchange of the inner polypyrrole film. The performance of the miniaturized DNA biosensor system was studied in respect to selectivity, sensitivity, reproducibility, and regeneration of the sensor. Control experiments were performed with a noncomplementary target of 27-mer DNA and 12 base-pair mismatched 18-mer sequences, respectively, and did not show any unspecific binding. Under optimized experimental conditions, the label-free electrochemical biosensor enabled the detection limits of 0.16 and 3.5 fmol for the 18- and 27-mer DNA strand, respectively. Furthermore, we demonstrate reusability of the electrochemical DNA biosensor after successful recovery of up to 100% of the original signal by regenerating the DNA "label-free" electrode with 50 mM HCl at room temperature. PMID- 16478106 TI - Comprehensive analysis of the substituent distribution in the glucosyl units and along the polymer chain of hydroxyethylmethyl celluloses and statistical evaluation. AB - Hydroxyethylmethyl celluloses (HEMC, DS(Me) 1.46-1.66, DS(HE) = 0.14-0.17) have been analyzed with respect to their methyl and hydroxyethyl pattern in the glucosyl units and along the polymer chain. Methyl groups were located by GLC/MS after direct hydrolysis, reduction, and acetylation, and the distribution of hydroxyethyl residues in the glucosyl units could be determined with enhanced sensitivity after permethylation to unify a certain HE pattern occurring in combination with various methyl patterns in a single peak. To get insight into the distribution of Me and HE along the cellulose chain, a method was developed which overcomes the strong discrimination of relative ion intensities caused by hydroxyalkyl groups and enables quantitative determination of the oligomer composition after random degradation for the first time. This comprises perdeuteriomethylation; partial acid hydrolysis; reductive amination with propylamine; and, finally, permethylation to yield completely O- and N-alkylated, permanently charged oligosaccharides. Although the methyl pattern can be determined by electrospray ionization ion-trap mass spectrometry (ESI-IT-MS) and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), as well, only MALDI-TOF-MS produced representative data for a quantitative evaluation of the HE pattern. Distribution of HE groups matches with a random distribution calculated from the monomer composition, whereas the methyl pattern was heterogeneous to a different extent. PMID- 16478107 TI - Quartz crystal microbalance studies of polymer gels and solutions in liquid environments. AB - Spreading of liquids and soft solids on a rigid surface in a surrounding liquid medium is investigated by utilizing the lateral sensitivity of the quartz crystal microbalance (QCM). While the QCM has been used extensively to study systems with spatial variations in the direction normal to the crystal's electrodes, few studies have exploited the QCM's ability to sense changes in loading in the plane of the electrodes. We propose equations to describe the predicted response of the QCM to a generalized viscoelastic material spreading at the QCM surface at the expense of the surrounding liquid medium. Several experimental examples are given in order to support the validity of the proposed equations, including situations where the spreading material is a Newtonian liquid, a viscoelastic liquid, or one of two viscoelastic solids. The first viscoelastic solid is a physically cross linked gel based on a styrene/ethylene-butene/styrene triblock copolymer in mineral oil, and the second is a cross-linked poly(ethylene glycol) hydrogel. PMID- 16478108 TI - Analytical evidence for the monolayer-protected cluster Au225[(S(CH2)5CH3)]75. AB - The Brust synthesis of thiolate-protected gold clusters has been modified to produce identifiable proportions of a hexanethiolate-protected Au225 core nanoparticle that display quantized double layer charging voltammetry consistent with a Au225 core dimension. Transmission electron microscopy (TEM) and thermogravimetric results indicate an average nanoparticle formula of Au225[(S(CH2)5CH3)]75. A simulated pulse voltammogram that accounts for the TEM nanoparticle dispersity matches reasonably well with that of the polydisperse synthetic sample containing the Au225 component. In confirmation of the size determination, an HPLC analysis using ratiometric absorbance and electrochemical detectors gives a core radius of 1.0 nm for the Au225 nanoparticle. PMID- 16478109 TI - In vitro and in situ tracking of choline-phospholipid biogenesis by MALDI TOF-MS. AB - The quantitative monitoring of newly synthesized species of phosphatidylcholines (PCs) and sphingomyelins (SMs) has been achieved in cultured human lens epithelial cells, both in situ and in vitro, with the use of MALDI TOF-MS. As the cells were cultured with deuterated choline-d(9), new peaks that differed from the hydrogenated species by 9.06 Da appeared in the mass spectra. The initial rates of appearance of all deuterated species of PCs were comparable and 4 times higher than those for SMs. After 12 h, those rates began to decrease for PCs but not for deuterated SMs, whose relative contents continued to increase throughout the 72 h of the experiment. The differences in initial rates are consistent with the reported initial generation of PCs, their subsequent degradation, and transfer of their headgroup, phosphorylcholine, to SMs. To further test the ability of MALDI TOF-MS to quantify changes in phospholipid (PL) metabolic pathways, myriocin, an inhibitor of SM synthesis, was added to the cells. In vitro and in situ results revealed a decrease in SMs and an unexpected increase in some PCs. With the use of other deuterated precursors and in combination with postsource decay or tandem MS/MS, this approach could allow the simultaneous tracking of the biosynthesis of multiple PL classes while providing details on their acyl chains. PMID- 16478110 TI - Method for characterizing sulfated glycoproteins in a glycosylation site-specific fashion, using ion pairing and tandem mass spectrometry. AB - Structural analysis of sulfated glycans is essential in understanding their biological significance. Here, we present a new approach to characterize sulfated glycans present on glycoproteins. The analysis is performed on glycopeptides, so information about the sulfated species is obtained in a glycosylation site specific manner. This method employs an ion-pairing reagent to stabilize the SO3 group of the glycopeptide, allowing useful information to be obtained during MS/MS experiments. The amount of structural information obtained from (+)ESI MS/MS of the ion-pair complexes for sulfated glycopeptides of equine thyroid stimulating hormone is compared with information obtained by (-)ESI-MS/MS of the underivatized, sulfated glycopeptides. The results indicate that this new method provides detailed insights into the sequence, branching, and type of N-glycans present, compared to analysis via (-)ESI-MS/MS. PMID- 16478111 TI - Comparison of atmospheric pressure photoionization, atmospheric pressure chemical ionization, and electrospray ionization mass spectrometry for analysis of lipids. AB - In this work, we compare the quantitative accuracy and sensitivity of analyzing lipids by atmospheric pressure photoionization (APPI), atmospheric pressure chemical ionization (APCI), and electrospray ionization (ESI) LC/MS. The target analytes include free fatty acids and their esters, monoglyceride, diglyceride, and triglyceride. The results demonstrate the benefits of using LC/APPI-MS for lipid analysis. Analyses were performed on a Waters ZQ LC/MS. Normal-phase solvent systems were used due to low solubility of these compounds in aqueous reversed-phase solvent systems. By comparison, APPI offers lower detection limits, generally highest signal intensities, and the highest S/N ratio. APPI is 2-4 times more sensitive than APCI and much more sensitive than ESI without mobile-phase modifiers. APPI and APCI offer comparable linear range (i.e., 4-5 decades). ESI sensitivity is dramatically enhanced by use of mobile phase modifiers (i.e., ammonium formate or sodium acetate); however, these ESI adduct signals are less stable and either are nonlinear or have dramatically reduced linear ranges. Analysis of fish oils by APPI shows significantly enhanced target analyte intensities in comparison with APCI and ESI. PMID- 16478112 TI - Cathodic detection of H2O2 using iodide-modified gold electrode in alkaline media. AB - Oxidative chemisorption and cathodic stripping reductive desorption of iodide have been studied at a smooth polycrystalline gold (Au (poly)) electrode. Potential-dependent surface coverage of iodide has been controlled on the basis of its reductive desoprtion in 0.1 M KOH alkaline media and its quantitative oxidation to aqueous iodates in acidic media. The Au (poly) electrode surface catalyzes the decomposition of H2O2 to O2. Specific adsorption of iodide on the Au electrode inhibits fully the catalytic decomposition and electrochemical oxidation of H2O2 as well as the adsorption of unknown impurities and the oxidative degradation of the electrode surface by H2O2. A quantitative characterization/detection of H2O2 at the iodide-modified Au (poly) electrode in the alkaline media has, thus, been achieved. Performance of the electrode toward the detection of H2O2 with respect to response time and sensitivity as well as operational stability has been evaluated. It has a sensitivity of 0.272 mA cm(-2) mM(-1) in amperometric measurements with a detection limit of 1.0 x 10(-5) M H2O2, and the response time to achieve 95% of the steady-state current is <20 s. The effect of O2 in the air-saturated solution can be minimized by subtracting the additional current for the O2 reduction. Experimental measurements were based upon cyclic voltametric and amperometric techniques. PMID- 16478113 TI - Development of a compound-specific carbon isotope analysis method for atmospheric formaldehyde via NaHSO3 and cysteamine derivatization. AB - A novel method has been developed for the compound-specific carbon isotope analysis of atmospheric formaldehyde using gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS). The method allows the determination of the delta13C value for atmospheric formaldehyde at nanogram levels with higher precision and lower detection limit. In the present work, atmospheric formaldehyde was collected using NaHSO3-coated Sep-Pak silica gel cartridges, washed out by water, then derivatized by cysteamine of known delta13C value, and the delta13C value of its derivative (thiazolidine) determined by GC/C/IRMS. Finally, the delta13C value of atmospheric formaldehyde could be calculated by a simple mass balance equation between formaldehyde, cysteamine, and thiazolidine. Using three formaldehydes with different delta13C values, calibration experiments were carried out over large ranges of formaldehyde concentrations. The carbon isotope analysis method achieved excellent reproducibility and high accuracy. There was no carbon isotopic fractionation throughout the derivatization processes. The differences in the carbon isotopic compositions of thiazolidine between the measured and predicted values were always <0.5 per thousand, within the specifications of the GC/C/IRMS system. The present method was also compared with the previous 2,4-dinitrophenylhydrazine derivatization method, and this method could be performed with lower analytical error and detection limit. Using this method, four 6-h ambient atmospheric formaldehyde samples were consecutively collected from 8 to 9 March 2005. The results showed that the delta13C values of atmospheric formaldehyde were different during the daytime and nighttime. This method proved suitable for the routine operation and may provide additional insight on sources and sinks of atmospheric formaldehyde. PMID- 16478114 TI - Supercritical fluid chromatography-tandem mass spectrometry for the enantioselective determination of propranolol and pindolol in mouse blood by serial sampling. AB - Packed-column supercritical fluid chromatography (pSFC) coupled to an atmospheric pressure chemical ionization (APCI) source and a tandem mass spectrometer (MS/MS) with minimum sample pretreatment was explored for the rapid and enantioselective determination of (R,S)-propranolol in mouse blood. Serial bleeding of mice is advantageous for the reduction of animal usage, dosing errors, and animal-to animal variation. The effects of the eluent flow rate and composition as well as the nebulizer temperatures on the ionization efficiency of racemic propranolol and pindolol as model compounds in the positive ion mode under pSFC conditions were studied. The fundamental parameters on the proposed hyphenated system such as matrix ionization suppression and chromatographic performances were investigated in improving sensitivity and enantiomeric separation for the detection of the analytes. The proposed chiral pSFC-APCI/MS/MS approach requiring approximately 3 min/sample for the determination of (R,S)-propranolol at a low nanogram per milliliter region was partially validated with respect to specificity, linearity, reproducibility, and accuracy and was applied to support a pharmacokinetic study. PMID- 16478115 TI - Ion trap versus low-energy beam-type collision-induced dissociation of protonated ubiquitin ions. AB - The beam-type and ion trap collision-induced dissociation (CID) behaviors of protonated bovine ubiquitin ions were studied for charge states ranging from +6 to +12 on a modified triple quadrupole/linear ion trap tandem mass spectrometer. Both beam-type CID and ion trap CID were conducted in a high-pressure linear ion trap, followed by proton-transfer ion/ion reactions to reduce the charge states of product ions mostly to +1. The product ions observed under each activation condition were predominantly b- and y-type ions. Fragmentation patterns showed a much stronger dependence on parent ion charge state with ion trap CID than with beam-type CID using nitrogen as the collision gas, with preferential cleavages C terminal to aspartic acid at relatively low charge states, nonspecific fragmentation at moderate charge states, and favored cleavages N-terminal to proline residues at high charge states. In the beam-type CID case, extensive cleavage along the protein backbone was noted, which yielded richer sequence information (77% of backbone amide bond cleavages) than did ion trap CID (52% of backbone amide bond cleavages). Collision gas identity and collision energy were also evaluated in terms of their effects on the beam-type CID spectrum. The use of helium as collision gas, as opposed to nitrogen, resulted in CID behavior that was sensitive to changes in collision energy. At low collision energies, the beam type CID data resembled the ion trap CID data with preferential cleavages predominant, while at high collision energies, nonspecific fragmentation was observed with increased contributions from sequential fragmentation. PMID- 16478116 TI - Surface plasmon resonance study of g protein/receptor coupling in a lipid bilayer free system. AB - Surface plasmon resonance (SPR) spectroscopy is a technique to study protein protein interactions in real time; however, application of SPR spectroscopy for investigations of membrane receptors is difficult with respect to functional and uniform immobilization of receptors on a biosensor surface. In the current study, we developed a simple, direct, biosensor-based approach to monitor the molecular interactions between G protein transducin (Gt) and rhodopsin (Rho), a prototypical G protein-coupled receptor (GPCR). Detergent-solubilized dark adapted Rho was captured onto a biosensor surface via lectin interaction, enabling site-directed immobilization of the receptor that made its cytoplasmic surface accessible to a coupling G protein. The system resembled the natural system with respect to receptor density, binding of Gt following flash or constant light application, fast GTP-dependent dissociation of Gt from Rho, regeneration of Rho, and dependence of Gt binding on light intensity and on concentration of Gt. The apparent KD of the Gt/Rho interaction was 13.6 nM. Our results validate the use of SPR spectroscopy as a tool to study G protein activation in GPCR systems and could be extended for application to other interaction partners of GPCRs. PMID- 16478117 TI - SearchXLinks. A program for the identification of disulfide bonds in proteins from mass spectra. AB - We present the computer program SearchXLinks that analyzes mass spectra with the aim of identifying disulfide bonds and other modifications in proteins of known amino acid sequence. Disulfide bonds can be intra- or intermolecular. To decrease the number of false positives, the analysis of in-source decay and tandem mass spectra are coupled into the program. The steps taken during a SearchXLinks run are outlined, and the computational costs are discussed. The application of the program is illustrated by the analysis of data from recent studies on bovine ribonuclease A and bovine serum albumin. The software can be used free of charge on the Internet at http://www.searchxlinks.de. PMID- 16478118 TI - Chromatography with two mobile phases. AB - Experimental results for the investigation of chromatographic columns containing two mobile phases are presented. The eluent was composed of mixtures of methanol and carbon dioxide. The column was an uncoated fused-silica-lined stainless steel capillary column. At certain experimental conditions, the eluent divided into two phases, both of which moved through the column. The predominant component of the liquid phase was methanol whereas the gas phase was composed of at least 93 mol % CO2. The columns were studied over a range of feed compositions (45-95 mol % CO2), pressures (61-101 bar), and temperatures (30-100 degrees C). The compositions and densities of each phase were calculated from the Peng-Robinson equation of state. The residence times of the two mobile phases were determined by tracer pulse chromatography. The partition coefficients of a probe solute, benzene, were measured along with the retention times of neon and the total volume of the chromatographic column as a function of temperature, pressure, and stoichiometric feed composition. The calculated column volumes, that is the volume of the liquid and gas, were constant over the full range of feed composition. The partition coefficient of benzene was constant at fixed pressure and temperature, varied logarithmically with density at fixed temperature and feed composition, and displayed a maximum at intermediate temperatures at fixed pressure and feed composition. The measured retention times of neon were consistently equivalent to the calculated residence times of the gas phase, indicating that neon did not dissolve in the liquid phase and could thus serve as an accurate dead time marker. The implementation of chromatography with two mobile phases produces a chromatographic "window". There is a lower limit for the retention volume of all solutes, viz., the residence time of the gas phase, exactly the same as normal chromatography. However, elimination of the stationary phase produces an upper limit to the retention volumes of solutes. This upper limit is the residence time of the liquid phase, so there is a retention window such that tG < or = ti < or = tL for all solutes. PMID- 16478119 TI - Phosphate group-driven fragmentation of multiply charged phosphopeptide anions. Improved recognition of peptides phosphorylated at serine, threonine, or tyrosine by negative ion electrospray tandem mass spectrometry. AB - The nanoelectrospray product ion spectra of multiply charged phosphopeptide anions reveal the occurrence of phosphate-specific high-mass fragment ions of the type [M - nH - 79](n-1)-. These so far unrecognized fragments, which are observed for phosphoserine-, phosphothreonine-, and phosphotyrosine-containing peptides, are the counterparts of the established inorganic phosphopeptide marker ion found at m/z 79 = [PO3]-. The high-mass marker ions are formed with high efficiency at moderate collision offset values and are particularly useful for sensitive recognition of pSer-, pThr-, and pTyr-peptides due to the low background level in MS/MS spectra at m/z values above those of the precursor ions. By virtue of this feature, the detection of the new phosphorylation-specific fragment ions appears to be more sensitive than the detection of the low-mass phosphate marker ion at m/z 79, where a higher interference by nonspecific background signals is generally observed. The number of phosphate groups within a phosphopeptide can also be estimated on the basis of the [M - nH - 79](n-1)- ions, since these exhibit an effective, sequential neutral loss of H3PO4 of the residing phosphate groups. A mechanistic explanation for the formation of the [M - nH - 79](n-1)- ions from multiply charged phosphopeptides is given. The high-mass marker ions are proposed to originate from phosphopeptide anions, which carry two negative charges located at the phosphate group. A new search tool denominated "variable m/z gain analysis", which utilizes these newly recognized high-mass fragments for spotting of phosphopeptides in a negative ion parent scan, is proposed. The findings strengthen the value of negative ion ESI-MS/MS for analysis of protein phosphorylation. PMID- 16478120 TI - On-line back-extraction field-amplified sample injection method for directly analyzing cocaine and thebaine in the extractants by solvent microextraction. AB - This paper describes a novel method that applies on-line back-extraction field amplified sample injection (OLBE-FASI) to the extractants by solvent microextraction (SME) in capillary zone electrophoresis (CZE). To our knowledge, it provides the first report that the water-immiscible solvent samples were directly analyzed by CZE. The water-immiscible solvent sample, sealed with a water plug in the sample vial, was used for direct electroinjection. The water plug with a moderate content of organic solvent, low-conductivity, and the presence of a small amount of H+ provided the highest sensitivity for analyzing positively chargeable compounds, such as cocaine and thebaine. The linear range was at least 2 orders of magnitude, with the square of the correlation coefficient (r2) > 0.9999, and a separate calibration over the range 0.016-10 microg/mL showed the linear range to be approaching 3 orders of magnitude. Detection limits were in the range of 2-10 ng/mL. Because the need to perform solvent exchange (from organic to aqueous solution) was eliminated, the OLBE-FASI method could be conveniently coupled with SME. In the present work, SME-OLBE-FASI CE was validated for quantitative purposes, and applications to human urine were demonstrated. PMID- 16478121 TI - Detecting deamidation products in proteins by electron capture dissociation. AB - A nonenzymatic posttranslational modification of proteins and peptides is the spontaneous deamidation of asparaginyl residues via a succinimide intermediate to form a varying mixture of aspartyl and isoaspartyl residues. The isoaspartyl residue is generally difficult to detect particularly using mass spectrometry because isoaspartic acid is isomeric with aspartic acid so that there is no mass difference. However, electron capture dissociation has demonstrated the ability to differentiate the two isoforms in synthetic peptides using unique diagnostic ions for each form; the cr. + 58 and z(l-r) - 57 fragment ions for the isoAsp form and the Asp side chain loss ((M + nH)(n-1)+. - 60) for the Asp form. Shown here are three examples of isoaspartyl detection in peptides from proteins; a deamidated tryptic peptide of cytochrome c, a tryptic peptide from unfolded and deamidated ribonuclease A, and a tryptic peptide from calmodulin deamidated in its native state. In all cases, the cr. + 58 and z(l-r) - 57 ions allowed the detection and localization of isoaspartyl residues to positions previously occupied by asparaginyl residues. The (M + nH)(n-1)+. - 60 ions were also detected, indicating the presence of aspartyl residues. Observation of these diagnostic ions in peptides from proteins shows that the method is applicable to defining the isomerization state of deamidated proteins. PMID- 16478122 TI - Microbial metabolomics with gas chromatography/mass spectrometry. AB - An analytical method was set up suitable for the analysis of microbial metabolomes, consisting of an oximation and silylation derivatization reaction and subsequent analysis by gas chromatography coupled to mass spectrometry. Microbial matrixes contain many compounds that potentially interfere with either the derivatization procedure or analysis, such as high concentrations of salts, complex media or buffer components, or extremely high substrate and product concentrations. The developed method was extensively validated using different microorganisms, i.e., Bacillus subtilis, Propionibacterium freudenreichii, and Escherichia coli. Many metabolite classes could be analyzed with the method: alcohols, aldehydes, amino acids, amines, fatty acids, (phospho-) organic acids, sugars, sugar acids, (acyl-) sugar amines, sugar phosphate, purines, pyrimidines, and aromatic compounds. The derivatization reaction proved to be efficient (>50% transferred to derivatized form) and repeatable (relative standard deviations <10%). Linearity for most metabolites was satisfactory with regression coefficients better than 0.996. Quantification limits were 40-500 pg on-column or 0.1-0.7 mmol/g of microbial cells (dry weight). Generally, intrabatch precision (repeatability) and interbatch precision (reproducibility) for the analysis of metabolites in cell extracts was better than 10 and 15%, respectively. Notwithstanding the nontargeted character of the method and complex microbial matrix, analytical performance for most metabolites fit the requirements for target analysis in bioanalysis. The suitability of the method was demonstrated by analysis of E. coli samples harvested at different growth phases. PMID- 16478124 TI - Method for ultra-low-level analysis of gold in rocks. AB - A new method for analyzing gold at ultralow concentrations (<10 pg/g) in geological samples has been developed involving HF-aqua regia acid digestion, chromatographic separation of Au from matrix elements using DIBK supported on an inert resin, and analysis by inductively coupled plasma-mass spectrometry (ICPMS). This method has an analytical detection limit of 2 parts per trillion (pg/g), significantly lower than most routinely used methods developed for analysis of ore samples with Au concentrations considerably higher than average crustal abundance ( approximately 2 ng/g). Such methods commonly have detection limits in the low nanogram per gram range. Many areas of geological research including ore genesis, crustal mobility and redistribution, planetary differentiation, and plume volcanism require quantitative analysis of geological materials with much lower Au concentrations. We present a rapid, easy to use method where Au is separated from matrix elements onto extractant primed chromatographic resin and analyzed by quadrupole ICPMS. The method is suitable for the relatively rapid analysis of a large number of samples and is reliable over a wide range of concentrations from picogram to microgram per gram level. Analysis of four different geostandards, GXR1, GXR4, CH-3, and SARM 7, yields concentrations within error of the published concentrations with accuracies of >95%. PMID- 16478123 TI - Identification and characterization of isomeric intermediates in a catalyst formation reaction by means of speciation analysis using HPLC-ICPMS and HPLC-ESI MS. AB - Information on chemical speciation is much needed in mechanistic and kinetic studies on catalyst formation processes in pharmaceutical research. Speciation analysis was applied to the identification and quantification of various rhodium species involved in a ligand exchange process leading to formation of catalyst dirhodium(II) tetrakis[methyl 2-oxopyrrolidin-5(S)-carboxylate]. Inductively coupled plasma mass spectrometry (ICPMS) was used as an element-specific detector following species separation by reversed-phase high-performance liquid chromatography (RP-HPLC), and electrospray ionization mass spectrometry (ESI-MS) was used for species identification and confirmation. A novel interface between the HPLC and ICPMS, which consisted of an eluent splitter, a desolvation unit, and the ICPMS built-in peristaltic pump, enabled the use of RP-HPLC with gradient elution and up to 100% organic components in the LC eluent without organic loading in the plasma. A variety of reaction intermediates were identified and quantified along the pathway to formation of the desired product, including isomeric di-, tri-, and tetrasubstituted species previously believed to be absent. This has provided new insights into the mechanism and kinetics of the reaction. The combination of HPLC-ICPMS and HPLC-ESI-MS has proven to be a valuable tool for the investigation of species evolution in catalyst formation process. PMID- 16478125 TI - Metabonomics and biomarker discovery: LC-MS metabolic profiling and constant neutral loss scanning combined with multivariate data analysis for mercapturic acid analysis. AB - In the field of metabonomics, 1H NMR and full scan mass spectrometry methods have usually been combined with principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) to detect patterns in biofluids that correspond to specific effects, usually a toxic site effect of a compound. Confounders together with great interindividual variation complicate such analysis in humans, and therefore, metabonomic data are almost restricted to animals. In our study, a constant neutral loss (CNL) scan on a linear ion trap demonstrated increased sensitivity and specificity compared to a full scan approach and was performed to detect mercapturic acids (MA), a class of effect markers. The method was applied to human volunteers administered 50 and 500 mg of acetaminophen (AAP), a model compound known to form MAs. Using a new algorithm to prepare the CNL data for chemometrics, discrimination of control and postdose samples could be performed using PCA and PLS-DA. The loadings plots clearly revealed AAP-MA as a marker, even at low-dose levels. Orthogonal signal correction (OSC) was carried out to investigate background information that is not due to exposure. Surprisingly, the OSC data provided a classification of male and female subjects showing the performance of the new approach. PMID- 16478126 TI - Molecular adsorption to LiMo3Se3 nanowire film chemiresistors. AB - Thin films of metallic nanowire bundles derived from the Chevrel compound LiMo3Se3 undergo reversible increases of their electrical resistance (up to 70%) upon exposure to vapors of organic solvents (Qi, X. B.; Osterloh, F. E. J. Am. Chem. Soc. 2005, 127 (21), 7666-7667). Using quartz crystal microbalance measurements with four analytes, we demonstrate here that the temporal and steady state resistance changes of the films depend on the time following the adsorption and on the number of molecules that adsorb to the nanowire films at a given pressure. The adsorption ability of the films and the corresponding film resistance increase in the row: hexane < THF < ethanol < DMSO, closely following the polarities of the solvents. On average, approximately 10(5) analyte molecules per LiMo3Se3 unit are required to produce a measurable electrical response. Atomic force microscopy scans on nanowire films reveal that analytes deposit on top of the nanowire bundles and cause the films to swell by approximately 6% in volume. The temporal and steady-state resistance data of the LiMo3Se3 chemiresistors can be explained by assuming that coating of the nanowire bundles with analyte molecules reduces the interwire charge transport in the films. PMID- 16478127 TI - Acetonitrile covalent adduct chemical ionization mass spectrometry for double bond localization in non-methylene-interrupted polyene fatty acid methyl esters. AB - Covalent adduct chemical ionization (CACI) using a product of acetonitrile self reaction, (1-methyleneimino)-1-ethenylium (MIE; CH2=C=N+=CH2), has been investigated as a method for localizing double bonds in a series of 16 non methylene-interrupted fatty acid methyl esters (NMI-FAME) of polyenes with three and more double bonds. As with polyunsaturated homoallylic (methylene interrupted) FAME and conjugated dienes, MIE (m/z 54) reacts across double bonds to yield molecular ions 54 mass units above the parent analyte. [M + 54]+ ions of several 20- and 22-carbon FAME that include one double bond in the C2-C3 position separated by two to five methylene units from a three, four, or five C homoallylic system dissociated according to rules for the homoallylic system, with an additional fragment corresponding to cleavage between the lone double bond and the carboxyl group and defining the position of the lone double bond. Triene FAME with both methylene and ethylene interruption yielded characteristic fragments distinguishable from homoallylic trienes. Fragmentation of fully conjugated trienes in the MS-1 spectra yields ratios of [M + 54]+/[M + 54 - 32]+ (loss of methanol) near unity, which distinguishes them from homoallylic FAME having a ratio of 8 or more; collisionally activated dissociation of [M + 54]+ yields a series of ions, including some rearrangement products, indicative of double bond position. Unlike conjugated dienes, fully conjugated triene diagnostic ion signal ratios did not follow any pattern based on double bond geometry. Partially conjugated trienes behave similarly to monoenes and conjugated dienes, yielding [M + 54]+/[M + 54 - 32]+ of 2-3 and, permitting them to be assigned as partially conjugated FAME using the MS-1 spectrum. They yield unique MS/MS spectra with weaker but assignable fragment ions, along with a diagnostic fragment that locates the lone double bond and permits 6,10,12 octatrienoate to be distinguished from 6,8,12-octatrienoate. The presence of a triple bond did not affect fragment formation in a methylene-interrupted yne-ene but did change fragments in a conjugated yne-ene. These data extend the principle of double bond localization by acetonitrile CACI-MS/MS to double bond structure in complex FAME found in nature. PMID- 16478128 TI - Solid contact potentiometric sensors for trace level measurements. AB - A simple procedure for the development of a range of polymeric ion-selective electrodes (ISEs) with low detection limits is presented. The electrodes were prepared by using a plasticizer-free methyl methacrylate-decyl methacrylate copolymer as membrane matrix and poly(3-octylthiophene) as intermediate layer deposited by solvent casting on gold sputtered copper electrodes as a solid inner contact. Five different electrodes were developed for Ag+, Pb2+, Ca2+, K+, and I , with detection limits mostly in the nanomolar range. In this work, the lowest detection limits reported thus far with solid contact ISEs for the detection of silver (2.0 x 10(-9) M), potassium (10(-7) M), and iodide (10(-8) M) are presented. The developed electrodes exhibited a good response time and excellent reproducibility. PMID- 16478129 TI - Fabrication of internally tapered capillaries for capillary electrochromatography electrospray ionization mass spectrometry. AB - In this study, we report a novel procedure for fabricating internally tapered capillary columns suitable for the coupling of capillary electrochromatography (CEC) to electrospray mass spectrometry (ESI-MS). The internal tapers were prepared by slowly heating the capillary end in a methane/O2 flame. Due to continuous self-shrinking of the inner channel of the capillary, the inside diameter of the opening was reduced to 7-10 microm. The procedure is easy to handle, with no requirement for expensive equipment as well as elimination of problematic grinding of the tip. Several advantages of these new internal tapers, as compared to using externally tapered columns, are described. First, the problems of poor durability and tip breakage associated with external tapering were successfully overcome with the internal taper. A comparison of the online CEC/ESI-MS between external versus internal tapers showed that the latter provides enhanced electrospray stability, resulting in significantly lower short term noise and very short-term noise values. In turn, the more rugged design of internal tapers allows performing CEC/MS utilizing a harsh polar organic mobile phase, which was not previously successful using an external taper due to higher operating current and electrospray arcing. Next, data on the reproducibility of the internally tapered CEC/MS column using warfarin and beta-blockers as model analytes are presented. For example, when comparing the reproducibility for separation of warfarin under reversed-phase conditions, the internal taper demonstrated superior intraday % RSD (1.6-3.4) as compared to the external taper intraday % RSD (5-6). Last, the applicability of performing quantitative CEC/MS with internally tapered capillaries is demonstrated for simultaneous enantioseparation of beta-blockers. Impressive quantitative results include good linearity of calibration curves (e.g., R2 = 0.9940-0.9988) and limit of detection as low as 30 nM. The sensitive detection of a minor impurity of one enantiomer at the 0.1% level in a major chiral entity buttresses the suitability of compliance with FDA guidelines. PMID- 16478130 TI - Method for internal standard introduction for quantitative analysis using on-line solid-phase extraction LC-MS/MS. AB - A novel approach for on-line introduction of internal standard (IS) for quantitative analysis using LC-MS/MS has been developed. In this approach, analyte and IS are introduced into the sample injection loop in different steps. Analyte is introduced into the injection loop using a conventional autosampler (injector) needle pickup from a sample vial. IS is introduced into the sample injection loop on-line from a microreservoir containing the IS solution using the autosampler. As a result, both analyte and IS are contained in the sample loop prior to the injection into the column. Methodology allowed to reliably introduce IS and demonstrated injection accuracy and precision comparable to those obtained using off-line IS introduction (i.e., IS and analyte are premixed before injection) while maintaining chromatographic parameters (i.e., analyte and IS elution time and peak width). This new technique was applied for direct analysis of model compounds in rat plasma using on-line solid-phase extraction (SPE) LC MS/MS quantification. In combination with on-line SPE, IS serves as a surrogate IS and compensates for signal variations attributed to sample preparation and instrumentation factors including signal suppression. The assays yielded accuracy (85-119%), precision (2-16%), and analyte recovery comparable to those obtained using off-line IS introduction. Furthermore, on-line IS introduction allows for nonvolumetric sample (plasma) collection and direct analysis without the need of measuring and aliquoting a fixed sample volume prior to the on-line SPE LC-MS/MS analysis. Therefore, this methodology enables direct sample (plasma) analysis without any sample manipulation and preparation. PMID- 16478131 TI - Quantitative comparison of proteomic data quality between a 2D and 3D quadrupole ion trap. AB - A 2D ion trap has a greater ion trapping efficiency, greater ion capacity before observing space-charging effects, and a faster ion ejection rate than a traditional 3D ion trap mass spectrometer. These hardware improvements should result in a significant increase in protein identifications from complex mixtures analyzed using shotgun proteomics. In this study, we compare the quality and quantity of peptide identifications using data-dependent acquisition of tandem mass spectra of peptides between two commercially available ion trap mass spectrometers (an LTQ and an LCQ XP Max). We demonstrate that the increased trapping efficiency, increased ion capacity, and faster ion ejection rate of the LTQ results in greater than 5-fold more protein identifications, better identification of low-abundance proteins, and higher confidence protein identifications when compared with a LCQ XP Max. PMID- 16478132 TI - Application of ceramic carbon materials for solid-phase extraction of organic compounds. AB - Ceramic carbon materials were developed as new sorbents for solid-phase extraction of organic compounds using chlorpromazine as a representative. The macroporosity and heterogeneity of ceramic carbon materials allow extracting a large amount of chlorpromazine over a short time. Thus, the highly sensitive and selective determination of chlorpromazine in urine sample was achieved by differential pulse voltammograms after only 1-min extraction. The total analysis time was less than 3 min. In comparison with other electrochemical and electrochemiluminescent methods following 1-min extraction, the proposed method improved sensitivity by about 2 and 1 order of magnitude, respectively. The fast extraction, diversity, and conductivity of ceramic carbon materials make them promising sorbents for various solid-phase extractions, such as solid-phase microextraction, thin-film microextraction, and electrochemically controlled solid-phase extraction. The preliminary applications of ceramic carbon materials in chromatography were also studied. PMID- 16478133 TI - Chiral ionic liquid that functions as both solvent and chiral selector for the determination of enantiomeric compositions of pharmaceutical products. AB - We have successfully synthesized both enantiomers of a novel chiral ionic liquid, (R)- and (S)-[(3-chloro-2-hydroxypropyl) trimethylammonium][bis((trifluoromethyl)sulfonyl)amide] ((R)- and (S) [CHTA]+[Tf2N]-) in optically pure form by a simple ion exchange reaction from corresponding chloride salts that are commercially available. In addition to the ease of preparation, this chiral IL has relatively high thermal stability (up to 300 degrees C), is liquid at room temperature (glass transition temperature of 58.4 degrees C), and exhibits strong enantiomeric recognition. The high solubility power and strong enantiomeric recognition ability make it possible to use this chiral IL to solubilize an analyte and to induce diastereomeric interactions for the determination of enantiomeric purity. In fact, we have successfully developed a novel method based on the near-infrared technique with this chiral IL serving both as solvent and as a chiral selector for the determination of enantiomeric purity. Enantiomeric compositions of a variety of pharmaceutical products and amino acids with different shape, size, and functional groups can be sensitively (milligram concentration) and accurately (enantiomeric excess as low as 0.6%) determined by use of this method. PMID- 16478134 TI - Microfluidic particle sorter employing flow splitting and recombining. AB - This paper describes an improved microfluidic device that enables hydrodynamic particle concentration and size-dependent separation to be carried out in a continuous manner. In our previous study, a method for hydrodynamic filtration and sorting of particles was proposed using a microchannel having multiple branch points and side channels, and it was applied for continuous concentration and separation of polymer particles and cells. In the current study, the efficiency of particle sorting was dramatically improved by geometrically splitting fluid flow from a main stream and recombining. With these operations, particles with diameters larger than a specific value move toward one sidewall in the mainstream. This control of particle positions is followed by the perfect particle alignment onto the sidewall, which increases the selectivity and recovery rates without using a liquid that does not contain particles. In this study, a microchannel having one inlet and five outlets was designed and fabricated. By simply introducing particle suspension into the device, concentrations of 2.1-3.0-microm particles were increased 60-80-fold, and they were collected independently from each outlet. In addition, it was demonstrated that the measured flow rates distributed into each side channel corresponded well to the theoretical values when regarding the microchannel network as a resistive circuit. PMID- 16478137 TI - Hydrophobic and hydrophilic yoctowells as receptors in water. AB - Hydrophilic yoctowells (volume = 10-24 L) with OEG walls are introduced, which entrap tobramycin, a tetraamino trisaccharide, in water with a binding constant of 107 M-1 but do not interact with cellobiose. This is in contrast to corresponding hydrophobic yoctowells, which kinetically immobilize cellobiose in water, but do not entrap tobramycin. PMID- 16478135 TI - Surface-enhanced Raman optical activity on adenine in silver colloidal solution. AB - We report the collection of Raman optical activity (ROA) spectra of adenine in silver colloidal solution, that is, surface-enhanced Raman optical activity (SEROA) using considerably shorter data acquisition times, reduced excitation power, and lower concentration, as compared to classical ROA measurements on molecules of biological interest so far reported in the literature. These improvements in experimental parameters for ROA measurements can be explained by enhanced Raman signals in the local optical fields of the silver nanoparticles and by at least 1 order of magnitude higher values for circular intensity differences (CIDs), as compared to classical ROA that has been suggested before and theoretically discussed in terms of large field gradients near a metal surface. The measured ROA effect for adenine can be understood in terms of adsorption-induced chirality in the prochiral molecules on the silver nanoparticles. Surface-enhanced Raman optical activity offers potential capabilities for sensitive, rapid, stereochemical characterization of basic building blocks of biopolymers, such as amino acids and nucleosides, as well as biologically active molecules, in particular, also for probing organization and self-assembling of such molecules on metal surfaces. PMID- 16478138 TI - Atomic-level description of amyloid beta-dimer formation. AB - All-atom simulations have been carried out on a monomer and dimer of the aggregation-prone fragment (16-22) of amyloid beta peptide, which is implicated in Alzheimer's disease. The replica exchange molecular dynamics method, which has been successfully applied to peptide folding, is utilized as a means to sample the configurational space with proper Boltzmann weighting so that the structural, motional, and thermodynamic description of self-assembly can be obtained. The free energy landscape showing the delicate balance between different monomer and dimer conformations is mapped along carefully chosen reaction coordinates. The canonical ensembles at 38 different temperatures are used to describe the thermodynamics and the relative stabilities of at least six different dimer conformations including that of parallel and antiparallel orientations. We also delineate the nature of the molecular forces that activate and stabilize these different dimer conformations as a function of temperature, especially as related to secondary structural propensity of monomer. We identify parallel loop dimer conformations that are stabilized due to specific interactions with water molecules. PMID- 16478139 TI - NMR distinction of single- and multiple-mode binding of small-molecule protein ligands. AB - Identifying and characterizing small-molecule inhibitors of protein-protein interactions is of high interest for drug discovery and for chemical genetics studies of biological pathways. Very often, initial hits or first-generation compounds have low micromolar dissociation constants and cause line broadening in NMR spectra. It is very important for subsequent structure-based compound optimization to know if this line broadening is caused by intermediate exchange of the dissociation kinetics only or in addition by multiple binding modes. Here, we present an approach of how to distinguish these two situations and demonstrate its experimental application. Two very similar small-molecule ligands of Bcl-xL are considered that cause both severe line broadening of interface residues. We show that one compound exhibits single-mode binding, and broadening is just due to dissociation kinetics in the intermediate exchange regime, and the line broadening can be overcome by providing excess ligand. In the other case, line broadening is due to dissociation kinetics and exchange between multiple bound conformations, and broadening cannot be overcome by providing excess ligand. The procedures used are very general and can also be applied to characterizing protein-protein and protein-nucleic acid interactions. PMID- 16478140 TI - Cytochrome display on amyloid fibrils. AB - Protein amyloid fibrils can be functionalized by coating the core protofilament with high concentrations of proteins and enzymes. This can be done elegantly by appending a functional domain to an amyloidogenic protein monomer, then assembling the monomers into a fibril. To display an array of biologically functional porphyrins on the surface of protein fibrils, we have fused the sequence of the small, soluble cytochrome b562 to an SH3 dimer sequence that can form classical amyloid fibrils rapidly under well-defined conditions. The resulting fusion protein also forms amyloid fibrils and, in addition, binds metalloporphyrins, at half of the porphyrin binding sites as shown by UV-vis and NMR spectroscopies. Once metalloporphyrins are bound to the fibrils, the resulting holo-cytochrome domains are spectroscopically identical to the wild type cytochrome. The concentration of metalloporphyrins on a saturated fibril is estimated to be of the order of approximately 20 mM, suggesting that they could be interesting systems for applications in nanotechnology. PMID- 16478141 TI - Paramagnetic active site models for the molybdenum-copper carbon monoxide dehydrogenase. AB - New paramagnetic, heterobimetallic Mo/Cu complexes featuring the Mo(=O)(mu-S)Cu core of O. carboxidovorans carbon monoxide dehydrogenase have been synthesized and structurally and spectroscopically characterized. The complexes exhibit EPR spectra (left graphic) indicative of extensive electron delocalization across the Mo-S-Cu core, in agreement with computational studies identifying the singly occupied molecular orbital (right graphic). PMID- 16478142 TI - Eight-membered ring construction by [4 + 2 + 2] annulation involving beta-carbon elimination. AB - Cyclobutanones underwent a formal [4 + 2 + 2] annulation reaction with 1,6- and 1,7-diynes in the presence of nickel(0) catalysts to provide bicyclic eight membered ring ketones. The annulation reaction proceeds through a ring-expansion of oxanickelacycloheptadiene via beta-carbon elimination to form a nine-membered nickelacycle. This reaction employing cyclobutanones as a C4 unit constructs cyclooctadienone cores in one synthetic step. PMID- 16478143 TI - Formation of nanostructured materials via coalescence of amphiphilic hollow particles. AB - A new, simplified route to amphiphilic core-shell nanotubes, microfibers, and microrods has been developed that does not involve the traditional utilization of well-defined block copolymers. Thus, amphiphilic graft copolymers (PEI-g-PMMA) are prepared by an aqueous free radical polymerization that self-assemble in situ to form uniform core-shell nanoparticles. The hydrophobic homopolymer (PMMA) that is also formed is incorporated in the cores. Slight cross-linking of the shells followed by extraction of the homopolymer results in hollow nanoparticles that coalesce to form nanotubes. When the shells are not cross-linked, the hollow particles coalesce to form microrods and microfibers. The sizes and shapes of the micromaterials can be controlled by varying the experimental conditions. PMID- 16478144 TI - 15N HYSCORE characterization of the fully deprotonated, reduced form of the archaeal Rieske [2Fe-2S] center. AB - The hyperfine couplings for strongly and weakly coupled 15N nuclei around a reduced Rieske [2Fe-2S] center of uniformly 15N-labeled, hyperthermostable archaeal Rieske protein at pH 13.3 were determined by hyperfine sublevel correlation (HYSCORE) spectroscopy and compared with those at physiological pH. Significant changes in the hyperfine couplings of the terminal histidine Ndelta ligands and Nepsilon nuclei were observed between them, which can be explained by not only the redistribution of the unpaired electron spin density over the ligands but also the difference in the mixed-valence state of the fully deprotonated, reduced cluster. These quantitative data can be used in theoretical analysis for the selection of an appropriate model of the mixed-valence state of the reduced Rieske center at very alkaline pH. PMID- 16478145 TI - MercuryII-mediated formation of thymine-HgII-thymine base pairs in DNA duplexes. AB - The very specific binding of the HgII ion unexpectedly and significantly stabilizes naturally occurring thymine-thymine base mispairing in DNA duplexes. Following this finding, we prepared DNA duplexes containing metal-mediated base pairs at the desired sites, as well as novel double helical architectures consisting only of thymine-HgII-thymine pairs. PMID- 16478146 TI - Enantioselective 1,3-dipolar cycloaddition reaction between diazoacetates and alpha-substituted acroleins: total synthesis of manzacidin A. AB - A titanium BINOLate catalyzed asymmetric 1,3-dipolar cycloaddition reaction between alpha-substituted acroleins and alkyl diazoacetates has been developed. With this methodology in hand, chiral 2-pyrazolines containing a quaternary stereogenic center were obtained in high to excellent enantioselectivities (up to 95% ee). The synthetic utility of the optically enriched 2-pyrazoline thus obtained was demonstrated in the short synthesis of manzacidin A. PMID- 16478147 TI - Stoichiometric hydrosilylation of nitriles with hydrido(hydrosilylene)tungsten complexes: formation of W-Si-N three-membered ring complexes and their unique thermal behaviors. AB - Reactions of hydrido(hydrosilylene)tungsten complexes, Cp'(CO)2(H)W=Si(H)[C(SiMe3)3], with nitriles (MeCN, tBuCN) at 60 degrees C gave hydrosilylation products, Cp'(CO)2W[kappa2(N,Si)-Si(H)(N=CHR'){C(SiMe3)3}] (R' = Me, tBu), with a novel W-Si-N three-membered ring structure. The product of the hydrosilylation of tBuCN underwent reversible rearrangement at 70 degrees C to a silylene complex, Cp'(CO)2(H)W=Si(N=CHtBu)[C(SiMe3)3], which was a major component in equilibrium. A reaction mechanism for the hydrosilylation involving coordination of nitriles to the silylene ligand and subsequent migration of the hydrido ligand to the nitrile carbon was proposed. PMID- 16478148 TI - Asymmetric synthesis of 4,5,6- and 3,4,5,6-substituted azepanes by a highly diastereoselective and enantioselective lithiation-conjugate addition sequence. AB - Asymmetric syntheses of 4,5,6- and 3,4,5,6-substituted azepanes have been achieved by highly diastereoselective and enantioselective (-)-sparteine-mediated asymmetric lithiation-conjugate additions of N-Boc-N-(p-methoxyphenyl)-2,3 substituted allylamines to a beta-aryl alpha,beta-unsaturated ester followed by hydrolysis, cyclization, and reduction. Access to the enantiomeric adduct is provided by an invertive lithiation-stannylation-lithiation sequence. PMID- 16478149 TI - A general and long-lived catalyst for the palladium-catalyzed coupling of aryl halides with thiols. AB - A general catalytic system for the coupling of aryl halides and sulfonates with thiols based on the use of the CyPF-t-Bu ligand (1) is reported. The reactions catalyzed by complexes of 1 occur in excellent yields with broad scope and exhibit extraordinary turnover numbers and high tolerance of functional groups. Turnover numbers usually exceed those of previous catalysts by 2 or 3 orders of magnitude. In addition, the reactions of aryl tosylates with alkane thiols to form aryl sulfides are reported for the first time. Finally, the synthesis of a diarylsulfide from two bromoarenes was accomplished using a hydrogen sulfide surrogate. PMID- 16478150 TI - Discovery of EGFR selective 4,6-disubstituted pyrimidines from a combinatorial kinase-directed heterocycle library. AB - The epidermal growth factor receptor (EGFR) tyrosine kinase was one of the first receptor tyrosine kinases to be targeted for drug development by the pharmaceutical industry due to its ubiquitous overexpression in a variety of tumors. Despite the validation of several quinazoline-based scaffolds in the clinic, there is a dearth of alternative chemical structure classes that are capable of inhibiting EGFR kinase activity selectively. Here we describe the discovery of potent and highly selective 4,6-disubstituted pyrimidine inhibitors of enzymatic and cellular EGFR activity and provide an explanation for their exceptional degree of kinase selectivity. PMID- 16478151 TI - Characterization of proton transport across a waveguide-supported lipid bilayer. AB - Cellular energy transduction processes are often driven by transmembrane ion gradients, and numerous artificial biomembrane systems have been developed that allow for chemically or light-induced charge transport into/out of liposomes. Liposomal architectures, however, are not readily interfaced to a solid-state transducer. Formation of an ion gradient across a planar-supported membrane, "wired" to a substrate electrode, may ultimately allow utilization of the potential energy to drive other electrochemical processes. Described here is a novel conductive polymer/planar waveguide assembly that provides for highly sensitive transduction of proton transport across a planar-supported lipid bilayer (PSLB). A quinone proton shuttle is embedded in the PSLB, which is coupled to the planar optical waveguide electrode through a pH-sensitive, self assembled conductive polymer film. Interfacial potential and absorbance changes in the conductive polymer film provide for sensitive characterization of transmembrane proton transport. The general and flexible nature of this architecture makes it adaptable to many different types of transmembrane transport chemistries, particularly light-activated systems. PMID- 16478152 TI - Catalytic "click" rotaxanes: a substoichiometric metal-template pathway to mechanically interlocked architectures. AB - A route to mechanically interlocked architectures that requires only a catalytic quantity of template is described. The strategy utilizes the Cu(I)-catalyzed 1,3 cycloaddition of azides with terminal alkynes. Chelating the Cu(I) to an endotopic-binding macrocycle means that the metal atom binds to the alkyne and azide in such a way that the metal-mediated bond-forming reaction occurs through the cavity of the macrocycle, forming a rotaxane. Addition of pyridine to the reaction mixture enables the Cu(I) to turn over during the reaction, permitting substoichiometric amounts of the metal to be used. The yields are very high for a rotaxane-forming reaction (up to 94% with stoichiometric Cu(I); 82% with 20 mol % of Cu(I)), and the procedure is practically simple to do (no requirement for an inert atmosphere nor dried or distilled solvents). PMID- 16478153 TI - Network single-walled carbon nanotube-field effect transistors (SWNT-FETs) with increased Schottky contact area for highly sensitive biosensor applications. AB - Highly sensitive single-walled carbon nanotube-field effect transistor (SWNT-FET) devices, which detect protein adsorptions and specific protein-protein interactions at 1 pM concentrations, have been achieved. The detection limit has been improved 104-fold compared to the devices fabricated by photolithography. The substantially increased sensitivity is mainly due to the increased Schottky contact area which accommodates relatively more numbers of proteins even at very low concentration. The augmented number of proteins adsorbed on a device induces instant modulation of the work function of metal contact electrodes, which eventually changes the conductance of the device. Such devices have been attained by addressing metal electrodes on network-type SWNTs using a shadow mask on a tilted angle sample stage. The shadow mask allows metals to penetrate underneath the mask efficiently, therefore forming a thin and wide Schottky contact area on SWNT channels. PMID- 16478154 TI - A comprehensive theoretical view of the bonding in actinide molecular complexes. AB - While usual atomic population methods give a consistent view of trivalent lanthanide or a uranium-ligand bond, the description of the bonding of americium(III), which is a key element for nuclear fuel processes, is a challenge. Neither experimental data nor theoretical calculations have been able so far to evidence covalency effects in the americium-ligand bond. We show herein that the use of more sophisticated methods based on a topological approach (AIM and ELF) gives a consistent view of such an interaction for the first time, showing a weak covalent back-bonding interaction with the CO ligand. PMID- 16478155 TI - PC bond cleavage of (silox)3NbPMe3 (silox = tBu3SiO) under dihydrogen leads to (silox)3Nb=CH2, (silox)3Nb=PH or (silox)3NbP(H)Nb(silox)3, and CH4. AB - Photolysis of the equilibrium mixture (silox)3NbPMe3 (1) + H2 (1-3 atm) right arrow over left arrow (silox)3Nb(Heq)2 (2e, tbp)/(silox)3Nb(Ht)2 (2t, pseudo-Td) + PMe3 causes PC bond cleavage. Depending on conditions, various amounts of (silox)3Nb=CH2 (3), (silox)3Nb=PH (5-H), (silox)3Nb=PMe (5-Me), (silox)3Nb=P(H)Nb(silox)3 (9, precipitated if N2 is present; X-ray), (silox)3NbH (4, active only through equilibration with 2e,t), and CH4 are produced. Addition of PH3 to 1 provides an independent route to 5-H; its deprotonation gives [(silox)3NbP]Li (6), whose methylation yields 5-Me. Early conversion 3:5-H ratios of approximately 3:1 suggest that initial PC bond activation is slow relative to subsequent PC bond cleavages. Addition of HPMe2 and H2PMe to 1 generates (silox)3HNbPMe2 (7) and (silox)3HNbPHMe (8), respectively, and both degrade faster than PMe3. A mechanism based around sequential PC or CH oxidative addition, followed by 1,2-elimination events, is proposed. The limiting step in the decomposition of all PMe3 is a slow hydrogenation of 3 to regenerate 2e,t and produces CH4. Hydrides 2e,t are likely to be the photolytically active species. PMID- 16478156 TI - Aggregation-induced folding of a de novo designed polypeptide immobilized on gold nanoparticles. AB - This communication reports the first steps in the construction of a novel, nanoparticle-based hybrid material for biomimetic and biosensor applications. Gold nanoparticles were modified with synthetic polypeptides to enable control of the particle aggregation state in a switchable manner, and particle aggregation was, in turn, found to induce folding of the immobilized peptides. PMID- 16478157 TI - Random coils, beta-sheet ribbons, and alpha-helical fibers: one peptide adopting three different secondary structures at will. AB - To potentially cure neurodegenerative diseases, we need to understand on a molecular level what triggers the complex folding mechanisms and shifts the equilibrium from functional to pathological isoforms of proteins. The development of small peptide models that can serve as tools for such studies is of paramount importance. We describe the de novo design and characterization of an alpha helical coiled coil based model peptide that contains structural elements of both alpha-helical folding and beta-sheet formation. Three distinct secondary structures can be induced at will by adjustment of pH or concentration. Low concentrations at pH 4.0 yield globular particles of the unfolded peptide, while at the same pH, but at higher concentration, defined beta-sheet ribbons are formed. In contrast, at high concentrations and pH 7.4, the peptide forms highly ordered alpha-helical fibers. Thus, this system allows one to systematically study now the consequences of the interplay between peptide and protein primary structure and environmental factors for peptide and protein folding on a molecular level. PMID- 16478158 TI - 4f-5f heterotrimetallic complexes exhibiting electrochemical and magnetic communication. AB - Novel 4f-5f complexes of U(IV) and Th(IV) with Yb have been prepared using a terpyridyl-functionalized ketimide linking group. These 4f-5f heterotrimetallic complexes (C5Me5)2An[-N=C(CH2C6H5)(tpyYb(C5Me5)2)]2 (where An = Th, U) exist in Yb(II/III) valence equilibria and exhibit rich electrochemical behavior consistent with electronic coupling between the actinide and Yb(II/III)tpy*- moieties. The magnetic response of the uranium complex, studied qualitatively using a subtraction method, is indicative of a coupled magnetic state between the U(IV) and Yb(III)tpy*- groups at low temperatures. Both the electrochemical and magnetic data are in agreement and are consistent with differences in participation of the actinide valence orbitals in the ketimide bonding. PMID- 16478159 TI - Hot spots in silver nanowire bundles for surface-enhanced Raman spectroscopy. AB - We report a simple strategy for placing analyte molecules in hot spots between closely spaced nanowires leading to intense SERS enhancement. The results are highly reproducible from experiment to experiment likely because of the regularity of the SERS substrate, which consists of highly ordered and regular silver nanowires fabricated in porous aluminum oxide. Because the silver nanowires are sealed in the pores of PAO, this system is potentially immune to contamination until it is ready for use, at which point the alumina matrix is etched, thereby allowing the silver nanowires to collapse into bundles and form hot spots in the region of close contact between the nanowires, trapping the analyte in those junctions. PMID- 16478160 TI - Expanding the thioglycoligase strategy to the synthesis of alpha-linked thioglycosides allows structural investigation of the parent enzyme/substrate complex. AB - For the first time, the thioglycoligase strategy has been successfully applied to alpha-glycosidases. The alpha-thioglycoligases derived from the family 31 glycosidases, alpha-xylosidase from E. coli (YicI) and alpha-glucosidase from Sulfolobus solfataricus, catalyze thioglycoligase reactions using alpha-glycosyl fluorides and deoxythioglycosides as donors and acceptors, respectively, in yields up to 86%. In addition, we describe the Michaelis complex of YicI using one of the thioglycosides as a nonhydrolyzable substrate analogue and discuss the structural insights this yields into the specificity and mechanism of family 31 alpha-glycosidases and the molecular basis of an associated genetic disease. PMID- 16478161 TI - The ionothermal synthesis of cobalt aluminophosphate zeolite frameworks. AB - Ionothermal synthesis, the use of an ionic liquid as both solvent and structure directing agent, has been used to synthesize three different cobalt aluminophosphate zeolites. Two of the materials are isostructural with solids prepared previously and have the AEI and SOD framework types. SIZ-7 is a novel zeolite structures that is closely related to the family of 8-ring zeolites consisting of the MER, GIS, and PHI frameworks. Single-crystal X-ray diffraction of SIZ-7 indicates that the distribution of cobalt among the four possible tetrahedral sites is unequal. PMID- 16478162 TI - Four-membered group 13 metal(I) N-heterocyclic carbene analogues: synthesis, characterization, and theoretical studies. AB - The synthesis, spectroscopic and structural characterization of the monomeric, four-membered group 13 metal(I) heterocycles ([:M{eta2-N,N'-(Ar)NC(NCy2)N(Ar)}], M = Ga or In, Ar = C6H3Pri2-2,6) and an isomeric thallium complex are reported. Theoretical studies on these complexes, which are analogues of four-membered N heterocyclic carbenes, suggest they should act as good sigma-donor ligands. PMID- 16478164 TI - Palladium-catalyzed stereo- and regiospecific allylation of aryl halides with homoallyl alcohols via retro-allylation: selective generation and use of sigma allylpalladium. AB - Treatment of tertiary homoallyl alcohol with aryl halide under palladium catalysis resulted in the transfer of the allyl moiety of the homoallyl alcohol to aryl halide and yielded the corresponding cross-coupling product stereo- and regiospecifically. The transfer process includes retro-allylation, which proceeds via a conformationally regulated six-membered transition state. The retro allylation can be regarded as a method for the stereo- and regiospecific preparation of sigma-allylpalladium. PMID- 16478163 TI - A screening method for chiral selectors that does not require covalent attachment. AB - A high-throughput screening protocol is proposed for chiral selector discovery. It is modeled after the protocol for biological screening of candidate drugs from chemical libraries. The procedure works based on target distribution between an aqueous phase and an organic phase. The target may be a racemate or separate enantiomers. Screening for noncovalent intermolecular association between target and candidate selectors is carried out by partitioning experiments in the presence and absence of the candidate chiral selectors in the organic phase (plasticized poly(vinyl chloride)). The partition ratio measurement uses 96-well plates for high throughput. The feasibility of this approach is validated by working with a known target/chiral selector pair, N-(3,5-dinitrobenzoyl)-alpha phenylglycine and 2,2,2-trifluoro-1-(9-anthryl)ethanol. The validated protocol is applied to a small library of 12 cyclopropyl dipeptide isosteres. Eight bind the racemic target, econazole. Among them, one has measurable chiral selectivity. The advantage of the method is that it does not require the covalent attachment of either the analyte or the selector, and the required amount of the potential chiral selector is about 100 mug. PMID- 16478165 TI - Photogenerated hole mobility in DNA measured by time-resolved microwave conductivity. AB - We report the first direct observation of charge transport dynamics using time resolved microwave conductivity and transient absorption spectroscopy techniques on the photolysis of an anthraquinone-bound DNA complex. PMID- 16478166 TI - Kinase-catalyzed modification of gold nanoparticles: a new approach to colorimetric kinase activity screening. AB - Peptide-stabilized gold nanoparticles have been enzymatically biotinylated by a kinase-catalyzed reaction using biotin-ATP as a cosubstrate. Upon mixing with avidin-modified particles, solutions of biotinylated particles change color from red to blue, indicating aggregation of particles. On the basis of this reaction, we have developed a simple colorimetric test to monitor kinase inhibitor activity. PMID- 16478167 TI - Electronic control of amide cis-trans isomerism via the aromatic-prolyl interaction. AB - The cis-trans isomerization of prolyl amide bonds results in large structural and functional changes in proteins and is a rate-determining step in protein folding. We describe a novel electronic strategy to control cis-trans isomerization, based on the demonstration that interactions between aromatic residues and proline are tunable by aromatic electronics. A series of peptides of sequence TXPN, X = Trp, pyridylalanine, pentafluorophenylalanine, or 4-Z-phenylalanine derivatives (Z = electron-donating, electron-withdrawing, or electron-neutral substituents), was synthesized and Ktrans/cis analyzed by NMR. Electron-rich aromatic residues stabilized cis amide bond formation, while electron-poor aromatics relatively favored trans amide bond formation. A Hammett correlation between aromatic electronics and cis-trans isomerization was observed. These results indicate that the interaction between aromatic residues and proline, which is observed to stabilize cis amide bonds and is also a general stabilizing interaction ubiquitous in proteins and protein-protein complexes, is not stabilized exclusively by a classical hydrophobic effect. To a large extent, the aromatic prolyl interaction is driven and controllable by an electronic effect between the aromatic ring pi-electrons and the proline ring, consistent with a C-H-pi interaction as the key stabilizing force. The aromatic-prolyl interaction is electronically tunable by 0.9 kcal/mol and is enthalpic in nature. In addition, by combining aromatic ring electronics and stereoelectronic effects using 4 fluoroprolines, we demonstrate broad tuning (2.0 kcal/mol) of cis-trans isomerism in tetrapeptides. We demonstrate a simple tetrapeptide, TWflpN, that exhibits 60% cis amide bond and adopts a type VIa1 beta-turn conformation. PMID- 16478168 TI - Dendritic folate rosettes as ion channels in lipid bilayers. AB - The self-assembly of folate dendrimers into pi-stacked supramolecular rosettes is shown to produce ion channels in planar and spherical lipid bilayer membranes. The found ion channels are small, quite homogeneous, long-lived, ohmic, cation selective (Eisenman I), and blockable by the permeant cation. PMID- 16478169 TI - Formation of ordered mesoporous films from in situ structure inversion of azo polymer colloidal arrays. AB - This work shows that mesoporous polymeric films with spherical and elliptical pores can be obtained by in situ structure inversion of the azo polymer colloid arrays through selective interaction with solvent. The epoxy-based azo polymer contained both the pseudo-stilbene-type azo chromophores and the hydrophilic carboxyl groups. The colloidal spheres of the azo polymer were prepared by gradual hydrophobic aggregation of the polymeric chains in THF-H2O media, induced by a steady increase in the water content. Ordered 2D arrays of the hexagonally close-packed colloidal spheres were obtained by the vertical deposition method. After the solvent (THF) annealing, the ordered 2D arrays were directly transformed to mesoporous films through the sphere-pore inversion. Under the same condition, the 2D arrays composed of the ellipsoidal colloids, which were obtained by the irradiation of a polarized Ar+ laser beam on the colloidal sphere arrays, could be transformed to films with ordered elliptical pores. To our knowledge, this is the first example to demonstrate that mesoporous structures can be directly formed from the colloidal arrays of a homopolymer through structure inversion. This observation can shed new light on the nature of self assembly processes and provide a feasible approach to fabricate mesoporous structures without the infiltration-removal step. By exploring the photoresponsive properties of the materials, mesoporous film with special pore structure and properties can be expected. PMID- 16478170 TI - Tris(pyrone) chelates of Gd(III) as high solubility MRI-CA. AB - Two tripodal, hexadentate pyrone-based chelators have been prepared. These ligands form stable, soluble complexes with gadolinium(III). The complexes show aqueous stability comparable to that of [Gd(DTPA)]2- at pH 7.4. Evaluation by relaxometry shows that these complexes have two inner-sphere water molecules and a very fast water exchange rate. The solution behavior of these complexes suggests that they may be attractive as high relaxivity, next-generation magnetic resonance imaging contrast agents. PMID- 16478171 TI - A chemo-enzymatic route to enantiomerically pure cyclic tertiary amines. AB - Deracemization of racemic chiral tertiary amines has been achieved by combination of an enantioselective amine oxidase, obtained through directed evolution, and ammonia borane in a one-pot process. PMID- 16478172 TI - Contrast viscosity changes upon photoirradiation for mixtures of poly(acrylic acid)-based alpha-cyclodextrin and azobenzene polymers. AB - Polymer-polymer interactions were investigated for mixtures of a poly(acrylic acid) (pAA) carrying azobenzene (pC12Azo) and two kinds of pAA carrying alpha cyclodextrin (CD), in which CDs are attached to the main chain through the 3- and 6-positions in CD (p3alphaCD and p6alphaCD, respectively), using several techniques, such as viscosity and NMR measurements. Viscosity data exhibited contrast changes upon UV irradiation: thinning (p3alphaCD/pC12Azo) and thickening (p6alphaCD/pC12Azo). NOESY spectra confirmed that the contrast viscosity changes were ascribable to differences in how CD moieties interact with pC12Azo after photoisomerization of azobenzene moieties from trans to cis: dissociation of inclusion complexes (p3alphaCD/pC12Azo) and formation of interlocked complexes (p6alphaCD/pC12Azo). PMID- 16478175 TI - Ene reaction of arynes with alkynes. AB - Arynes, generated in situ from ortho-silylaryl triflates, undergo ene reaction with alkynes possessing propargylic hydrogen in the presence of KF/18-crown-6 in THF at room temperature to give substituted phenylallenes. Various terminal and internal alkynes as well as different arynes can be used to give the corresponding phenylallenes in good to moderate yields. The reaction of alkyne without propargylic hydrogen gave an acetylenic C-H addition product (a phenylalkyne) and a dehydro Diels-Alder product (a phenanthrene). PMID- 16478173 TI - Quantum-dot/aptamer-based ultrasensitive multi-analyte electrochemical biosensor. AB - The coupling of aptamers with the coding and amplification features of inorganic nanocrystals is shown for the first time to offer a highly sensitive and selective simultaneous bioelectronic detection of several protein targets. This is accomplished in a single-step displacement assay in connection to a self assembled monolayer of several thiolated aptamers conjugated to proteins carrying different inorganic nanocrystals. Electrochemical stripping detection of the nondisplaced nanocrystal tracers results in a remarkably low (attomole) detection limit, that is, significantly lower than those of existing aptamer biosensors. The new device offers great promise for measuring a large panel of disease markers present at ultralow levels during early stages of the disease progress. PMID- 16478174 TI - Radiosensitization by a modified nucleotide that produces DNA interstrand cross links under hypoxic conditions. AB - This paper describes the reactivity of a molecule that combines two desirable chemical processes into one molecule for the first time. Interstrand cross-links (ISCs) are an effective family of lesions produced by cytotoxic agents that target DNA. For instance, ISCs are the source of mitomycin C's cytotoxicity. Radiosensitizing agents are molecules that enhance DNA damage produced by ionizing radiation, especially under O2-deficient conditions. Phenyl selenide 1 is the first example of a modified nucleotide that can be incorporated in DNA by polymerases, which produces ISCs when DNA containing it is exposed to gamma radiolysis under O2-deficient conditions. These experiments suggest that 1 could be useful as a novel type of radiosensitizing agent. PMID- 16478176 TI - High-throughput enzymatic method for enantiomeric excess determination of O acetylated cyanohydrins. AB - The reaction yield and enantiomeric excess of O-acetylated cyanohydrin reaction products from a library of chiral catalysts can be analyzed by a three-step screening method. Alcohol dehydrogenase and NADH are used to analyze unreacted substrate. A lipase with absolute specificity converts one enantiomer to a quantifiable product before the remaining enantiomer is hydrolyzed with an unspecific esterase and quantified. PMID- 16478177 TI - NMR chemical shift powder pattern recoupling at high spinning speed and theoretical tensor evaluation applied to silk fibroin. AB - The NMR pulse sequence RAI (recoupling of anisotropy information) has been improved to obtain powder patterns at high MAS spinning speeds. The 2D iso-aniso experiment displays the static chemical shift spectra on the indirect dimension and the MAS spectra on the direct dimension; hence overlapping chemical shift tensor patterns can be well resolved. This efficient technique is applicable to compounds containing (13)C sp(3) (C(alpha), C(beta)) and sp(2) (C=O) sites with higher chemical shift (CS) anisotropy (CSA), and the reliability of the method was tested here on the (13)C chemical shift tensors of polycrystalline glycine, alanine, and serine. Subsequently, the same experiment was applied to the native silk protein fibroin from Bombyx mori, which consists mainly of these three amino acids. Molecular dynamics (MD) simulations of the silk II crystal structure of Takahashi et al. (Takahashi et al. Int. J. Biol. Macromol. 1999, 24, 127-138) were carried out to study the influence of motions on the chemical shift tensors. The (13)C chemical shift tensors were calculated using the bond polarization theory BPT on 200 structures created by an MD simulation. Very good agreement of the theoretical chemical shift anisotropy values with the experimental NMR results was obtained. The tensor orientations in the protein structure could thus be reliably derived. PMID- 16478178 TI - Total synthesis and structural elucidation of azaspiracid-1. Construction of key building blocks for originally proposed structure. AB - Syntheses of the three key building blocks (65, 98, and 100) required for the total synthesis of the proposed structure of azaspiracid-1 (1a) are described. Key steps include a TMSOTf-induced ring-closing cascade to form the ABC rings of tetracycle 65, a neodymium-catalyzed internal aminal formation for the construction of intermediate 98, and a Nozaki-Hiyama-Kishi coupling to assemble the required carbon chain of fragment 100. The synthesized fragments, obtained stereoselectively in both their enantiomeric forms, were expected to allow for the construction of all four stereoisomers proposed as possible structures of azaspiracid-1 (1a-d), thus allowing the determination of both the relative and absolute stereochemistry of the natural product. PMID- 16478179 TI - Total synthesis and structural elucidation of azaspiracid-1. Synthesis-based analysis of originally proposed structures and indication of their non-identity to the natural product. AB - The key building blocks (6, 7, and 8) for the intended construction of the originally proposed structures of azaspiracid-1, a potent marine-derived neurotoxin, were coupled and the products elaborated to the targeted compounds (1a,b) and their C-20 epimers (2 and 3). The assembly of the three intermediates was accomplished by a dithiane-based coupling reaction that united the C(1)-C(20) (7) and C(21)-C(27) (8) fragments, followed by a Stille-type coupling which allowed the incorporation of the C(28)-C(40) fragment (6) into the growing substrate. Neither of the final products (1a,b) matched the natural substance by TLC or (1)H NMR spectroscopic analysis, suggesting one or more errors in the originally proposed structure for this notorious biotoxin. PMID- 16478180 TI - Photoactivated h/d exchange in tyrosine: involvement of a radical anion intermediate. AB - The aromatic hydrogen nuclei of tyrosine are photochemically labile and exchange with deuterons in neutral D(2)O solution. The site meta to the ring hydroxyl substituent is preferentially deuterated, exhibiting a meta/ortho deuteration rate of approximately 4:1. In contrast with acid-catalyzed H/D exchange and with nearly all of the reported photoactivated H/D exchange studies, the UV-induced H/D exchange of tyrosine is optimal at pH 9 and is effectively quenched at acid pH. Photochemical H/D exchange is strongly stimulated by the alpha-amino group (the aromatic hydrogens of p-cresol are far less subject to exchange) and by imidazole or phosphate buffers. On the basis of the results obtained here and on the previously identified cyclohexadienyl radical (Bussandri, A.; van Willigen, H. J. Phys. Chem. A 2002, 106, 1524-1532), we conclude that the exchange reaction involves a radical intermediate and results from two distinct roles of tyrosine: (1) as a phototransducer of light energy into solvated electrons (e(aq)(-)), and (2) as an acceptor of an electron to create a radical anion intermediate which is rapidly protonated, yielding a neutral cyclohexadienyl radical. Regeneration of the tyrosine can occur via a bimolecular redox reaction of the cyclohexadienyl and phenoxyl radicals to yield a carbocation/phenoxide pair, followed by deprotonation of the carbocation. The oxidation step is pH dependent, requiring the deprotonated form of the cyclohexadienyl radical. The H/D exchange thus results from a cyclic one-electron (Birch) reduction/protonation/reoxidation (by phenoxyl radical)/deprotonation cycle. Consistent with these mechanistic conclusions, the aromatic hydrogens of tyrosine O-methyl ether are photochemically inert, but become labile in the presence of tyrosine at high pH. The deuteration rate of O-methyl tyrosine is lower than that of tyrosine and shows a preference for the ortho positions. This difference is proposed to result from a variation in the oxidation step, characterized by a preferential oxidation of a cyclohexadienyl resonance structure with the unpaired electron localized on the oxygen substituent. PMID- 16478181 TI - Monoliths of aligned silica-polypeptide hexagonal platelets. AB - Water soluble alpha-helical polypeptides were used to prepare silica coated hexagonal single crystal platelets in concentrated solutions. To our knowledge, there is no other instance where polymer single crystals, typically formed under high dilution, can be grown in a bulk material. This unprecedented self-assembly process relies on complex cooperative interactions where silica condensation mediates the growth of polypeptide crystals, which in turn template silica overgrowth. The helices were also used to align samples giving monoliths composed of highly oriented layers of platelets. Overall, this procedure allows preparation of composites with good structural order and complexity via a simple biomimetic process. PMID- 16478182 TI - Rewritable DNA microarrays. AB - Thiol-terminated single-stranded deoxyribonucleic acids (ssDNA) can be immobilized onto pulsed plasma deposited poly(allylmercaptan) surfaces via disulfide bridge chemistry and are found to readily undergo nucleic acid hybridization. Unlike other methods for oligonucleotide attachment to solid surfaces, this approach is shown to be independent of substrate material or geometry, and amenable to highly efficient rewriting. PMID- 16478183 TI - Stereochemistry of imine reduction by a hydroxycyclopentadienyl ruthenium hydride. AB - The stereochemistry of hydrogen transfer from [2,5-Ph(2)-3,4-Tol(2)(eta(5) C(4)COD)]Ru(CO)(2)D to N-aryl imines to give amine complexes was shown to be mostly trans stereospecific. Stereospecific hydrogen transfer is proposed to generate an amine and a coordinatively unsaturated ruthenium intermediate in close proximity. Coordination of the amine is proposed to occur faster than lone pair inversion of the amine. In contrast, hydrogen transfer to N-alkyl imines is stereorandom. It is proposed that stereochemistry is lost in part due to the reversibility of the hydrogen transfer being faster than amine coordination. PMID- 16478184 TI - A europium complex that selectively stains nucleoli of cells. AB - A europium complex selectively staining the nucleolus of NIH 3T3, HeLa, and HDF cells is reported. This complex possesses not only the advantage of the long lifetime of europium emission (0.3 ms), but also a chromophore that allows excitation at a relatively long wavelength (lambda(max) = 384 nm) and gives rise to an acceptable quantum yield (9%). The complex can be used both in live cell and fixed cell imaging, giving an average intracellular concentration on the order of 0.5 microM. Strong binding to serum albumin has been demonstrated by examination of the analogous gadolinium complex, studying relaxivity changes with increasing protein concentration. The intracellular speciation of the complex has been examined by circularly polarized emission spectroscopy and is consistent with the presence of more than one europium species, possibly protein bound. PMID- 16478185 TI - The 2,6-diisocyanoazulene motif: synthesis and efficient mono- and heterobimetallic complexation with controlled orientation of the azulenic dipole. AB - Synthesis of the remarkably air- and thermally stable 2,6-diisocyano-1,3 diethoxycarbonylazulene linker from 2-amino-1,3-diethoxycarbonylazulene in 57% cumulative yield was developed. Incorporation of the ester "arms" in the design of this first diisocyanoazulene bridge permitted fully controlled stepwise installation and complexation of its isocyano junction groups. The -CO(2)Et arms in 2,6-diformamido-1,3-diethoxycarbonylazulene effectively suppress the rate of dehydration of its 2-NHCHO end relative to that of the 6-NHCHO end leading to practically exclusive formation of 6-isocyano-2-formamido-1,3 diethoxycarbonylazulene upon treatment of the above diformamide with an equimolar amount of POCl(3). This crystallographically characterized 6-isocyano-2 formamidoazulene derivative was employed to access mono- and heterobimetallic complexes of the 2,6-diisocyanoazulene scaffold with controlled orientation of the azulenic dipole. A complete series of monometallic, homobimetallic, and isomeric heterobimetallic ([M] = M(CO)(5), M = Cr and/or W) complexes of the 2,6 diisocyanoazulene motif was isolated and studied by a variety of techniques, including X-ray crystallography. The metal-to-bridge charge transfer in mono- and dinuclear adducts of 2,6-diisocyanoazulene, the assignment of which was corroborated by time-dependent density functional theory calculations, occurs at a dramatically lower energy as compared to the analogous systems featuring the 1,4-diisocyanobenzene scaffold. Moreover, the metal-to-diisocyanide charge transfer exhibits a substantially greater red shift upon binucleation of the mononuclear [M(CO)(5)] adducts of the nonbenzenoid 2,6-diisocyanoazulene linker versus the 1,4-diisocyanobenzene bridge. PMID- 16478186 TI - One-dimensional transition metal-benzene sandwich polymers: possible ideal conductors for spin transport. AB - We investigate the electronic and magnetic properties of the proposed one dimensional transition metal (TM = Sc, Ti, V, Cr, and Mn) -benzene (Bz) sandwich polymers by means of density functional calculations. [V(Bz)](infinity) is found to be a quasi-half-metallic ferromagnet, and half-metallic ferromagnetism is predicted for [Mn(Bz)](infinity). Moreover, we show that stretching the [TM(Bz)](infinity) polymers could have dramatic effects on their electronic and magnetic properties. The elongated [V(Bz)](infinity) displays half-metallic behavior, and [Mn(Bz)](infinity) stretched to a certain degree becomes an antiferromagnetic insulator. The possibilities to stabilize the ferromagnetic order in [V(Bz)](infinity) and [Mn(Bz)](infinity) polymers at finite temperatures are discussed. We suggest that the hexagonal bundles composed by these polymers might display intrachain ferromagnetic order at finite temperatures by introducing interchain exchange coupling. PMID- 16478187 TI - CNT-CdTe versatile donor-acceptor nanohybrids. AB - Single wall carbon nanotubes (SWNT) and multiwall carbon nanotubes (MWNT) were linked to thioglycolic acid (TGA)-capped CdTe nanoparticles (NP) through electrostatic interactions producing photoactive superstructures. The novel nanohybrids were characterized both in the ground and excited states with specific accent on electron-transfer chemistry. In fact, both assays provide kinetic and spectroscopic evidence that support a partial transfer of charge density, with rapid formation of microsecond-lived radical ion pair states. Since nanotubes provide a quick transportation route of charge carriers to the electrode, we took this remarkable finding further and constructed photoelectrochemical cells. Photocurrents were generated through the implementation of CdTe and SWNT or MWNT, which serve as excited-state electron donor components and electron acceptors, respectively. PMID- 16478188 TI - Solution state structure determination of silicate oligomers by 29SI NMR spectroscopy and molecular modeling. AB - Evidence for nine new solution state silicate oligomers has been discovered by (29)Si NMR homonuclear correlation experiments of (29)Si-enriched samples. In addition to enhancing signal sensitivity, the isotopic enrichment increases the probability of the (29)Si-(29)Si two-bond scalar couplings that are necessary for the observation of internuclear correlations in 2-D experiments. The proposed assignments are validated by comparisons of experimental and simulated cross peaks obtained with high digital resolution. The internuclear connectivity indicated by the NMR data suggests that several of these oligomers can have multiple stereoisomers, including conformers and/or diastereomers. The stabilities of these oligomers and their possible stereoisomers have been investigated by electronic structure calculations. PMID- 16478189 TI - Large area liquid crystal monodomain field-effect transistors. AB - Butyl, hexyl, and decyl derivatives of the liquid-crystalline organic semiconductor 5,5' '-bis(5-alkyl-2-thienylethynyl)-2,2':5',2' '-terthiophene were synthesized and studied with respect to their structural, optical, and electrical properties. By means of an optimized thermal annealing scheme the hexyl and decyl compounds could be processed into self-assembled monodomain films of up to 150 mm in diameter. These were investigated with X-ray diffractometry, which revealed a clearly single-crystalline monoclinic morphology with lamellae parallel to the substrate. Within the lamellae the molecules were found to arrange with a tilt of about 50 degrees with the rubbing direction of the polyimide alignment layer. The resulting, close side-to-side packing was confirmed by measurements of the UV/vis absorption, which showed a dichroic ratio of 19 and indicated H-aggregation. AFM analyses revealed self-affinity in the surface roughness of the monodomain. The compounds showed bipolar charge transport in TOF measurements, with hole mobilities reaching up to 0.02 cm(2)/Vs and maximum electron mobilities around 0.002 cm(2)/Vs. The hexyl derivative was processed into large-area monodomain top gate field-effect transistors, which were stable for months and showed anisotropic hole mobilities of up to 0.02 cm(2)/Vs. Compared to multidomain bottom-gate transistors the monodomain formation allowed for a mobility increase by 1 order of magnitude. PMID- 16478190 TI - Solvent isotope effects on interfacial protein electron transfer in crystals and electrode films. AB - D(2)O-grown crystals of yeast zinc porphyrin substituted cytochrome c peroxidase (ZnCcP) in complex with yeast iso-1-cytochrome c (yCc) diffract to higher resolution (1.7 A) and pack differently than H(2)O-grown crystals (2.4-3.0 A). Two ZnCcP's bind the same yCc (porphyrin-to-porphyrin separations of 19 and 29 A), with one ZnCcP interacting through the same interface found in the H(2)O crystals. The triplet excited-state of at least one of the two unique ZnCcP's is quenched by electron transfer (ET) to Fe(III)yCc (k(e) = 220 s(-1)). Measurement of thermal recombination ET between Fe(II)yCc and ZnCcP+ in the D(2)O-treated crystals has both slow and fast components that differ by 2 orders of magnitude (k(eb)(1) = 2200 s(-1), k(eb)(2) = 30 s(-1)). Back ET in H(2)O-grown crystals is too fast for observation, but soaking H(2)O-grown crystals in D(2)O for hours generates slower back ET, with kinetics similar to those of the D(2)O-grown crystals (k(eb)(1) = 7000 s(-1), k(eb)(2) = 100 s(-1)). Protein-film voltammetry of yCc adsorbed to mixed alkanethiol monolayers on gold electrodes shows slower ET for D(2)O-grown yCc films than for H(2)O-grown films (k(H) = 800 s(-1); k(D) = 540 s(-1) at 20 degrees C). Soaking H(2)O- or D(2)O-grown films in the counter solvent produces an immediate inverse isotope effect that diminishes over hours until the ET rate reaches that found in the counter solvent. Thus, D(2)O substitution perturbs interactions and ET between yCc and either CcP or electrode films. The effects derive from slow exchanging protons or solvent molecules that in the crystal produce only small structural changes. PMID- 16478191 TI - Low-temperature protein dynamics: a simulation analysis of interprotein vibrations and the boson peak at 150 k. AB - An understanding of low-frequency, collective protein dynamics at low temperatures can furnish valuable information on functional protein energy landscapes, on the origins of the protein glass transition and on protein-protein interactions. Here, molecular dynamics (MD) simulations and normal-mode analyses are performed on various models of crystalline myoglobin in order to characterize intra- and interprotein vibrations at 150 K. Principal component analysis of the MD trajectories indicates that the Boson peak, a broad peak in the dynamic structure factor centered at about approximately 2-2.5 meV, originates from approximately 10(2) collective, harmonic vibrations. An accurate description of the environment is found to be essential in reproducing the experimental Boson peak form and position. At lower energies other strong peaks are found in the calculated dynamic structure factor. Characterization of these peaks shows that they arise from harmonic vibrations of proteins relative to each other. These vibrations are likely to furnish valuable information on the physical nature of protein-protein interactions. PMID- 16478192 TI - Specific and stable fluorescence labeling of histidine-tagged proteins for dissecting multi-protein complex formation. AB - Labeling of proteins with fluorescent dyes offers powerful means for monitoring protein interactions in vitro and in live cells. Only a few techniques for noncovalent fluorescence labeling with well-defined localization of the attached dye are currently available. Here, we present an efficient method for site specific and stable noncovalent fluorescence labeling of histidine-tagged proteins. Different fluorophores were conjugated to a chemical recognition unit bearing three NTA moieties (tris-NTA). In contrast to the transient binding of conventional mono-NTA, the multivalent interaction of tris-NTA conjugated fluorophores with oligohistidine-tagged proteins resulted in complex lifetimes of more than an hour. The high selectivity of tris-NTA toward cumulated histidines enabled selective labeling of proteins in cell lysates and on the surface of live cells. Fluorescence labeling by tris-NTA conjugates was applied for the analysis of a ternary protein complex in solution and on surfaces. Formation of the complex and its stoichiometry was studied by analytical size exclusion chromatography and fluorescence quenching. The individual interactions were dissected on solid supports by using simultaneous mass-sensitive and multicolor fluorescence detection. Using these techniques, formation of a 1:1:1 stoichiometry by independent interactions of the receptor subunits with the ligand was shown. The incorporation of transition metal ions into the labeled proteins upon labeling with tris-NTA fluorophore conjugates provided an additional sensitive spectroscopic reporter for detecting and monitoring protein protein interactions in real time. A broad application of these fluorescence conjugates for protein interaction analysis can be envisaged. PMID- 16478193 TI - Structural evolution in a hydrothermal reaction between Nb2O5 and NaOH solution: from Nb2O5 grains to microporous Na2Nb2O6.2/3H2O fibers and NaNbO3 cubes. AB - Niobium pentoxide reacts actively with concentrate NaOH solution under hydrothermal conditions at as low as 120 degrees C. The reaction ruptures the corner-sharing of NbO(7) decahedra and NbO(6) octahedra in the reactant Nb(2)O(5), yielding various niobates, and the structure and composition of the niobates depend on the reaction temperature and time. The morphological evolution of the solid products in the reaction at 180 degrees C is monitored via SEM: the fine Nb(2)O(5) powder aggregates first to irregular bars, and then niobate fibers with an aspect ratio of hundreds form. The fibers are microporous molecular sieve with a monoclinic lattice, Na(2)Nb(2)O(6).(2)/(3)H(2)O. The fibers are a metastable intermediate of this reaction, and they completely convert to the final product NaNbO(3) cubes in the prolonged reaction of 1 h. This study demonstrates that by carefully optimizing the reaction condition, we can selectively fabricate niobate structures of high purity, including the delicate microporous fibers, through a direct reaction between concentrated NaOH solution and Nb(2)O(5). This synthesis route is simple and suitable for the large-scale production of the fibers. The reaction first yields poorly crystallized niobates consisting of edge-sharing NbO(6) octahedra, and then the microporous fibers crystallize and grow by assembling NbO(6) octahedra or clusters of NbO(6) octahedra and NaO(6) units. Thus, the selection of the fibril or cubic product is achieved by control of reaction kinetics. Finally, niobates with different structures exhibit remarkable differences in light absorption and photoluminescence properties. Therefore, this study is of importance for developing new functional materials by the wet-chemistry process. PMID- 16478194 TI - Quantum dot solar cells. harvesting light energy with CdSe nanocrystals molecularly linked to mesoscopic TiO2 films. AB - By using bifunctional surface modifiers (SH-R-COOH), CdSe quantum dots (QDs) have been assembled onto mesoscopic TiO(2) films. Upon visible light excitation, CdSe QDs inject electrons into TiO(2) nanocrystallites. Femtosecond transient absorption as well as emission quenching experiments confirm the injection from the excited state of CdSe QDs into TiO(2) nanoparticles. Electron transfer from the thermally relaxed s-state occurs over a wide range of rate constant values between 7.3 x 10(9) and 1.95 x 10(11) s(-1). The injected charge carriers in a CdSe-modified TiO(2) film can be collected at a conducting electrode to generate a photocurrent. The TiO(2)-CdSe composite, when employed as a photoanode in a photoelectrochemical cell, exhibits a photon-to-charge carrier generation efficiency of 12%. Significant loss of electrons occurs due to scattering as well as charge recombination at TiO(2)/CdSe interfaces and internal TiO(2) grain boundaries. PMID- 16478195 TI - Reactivity and selectivity in the Wittig reaction: a computational study. AB - The salt-free Wittig reaction of non-, semi-, and stabilized ylides has been investigated on realistic systems using density functional theory (DFT) calculations, including continuum solvation. Our results provide unequivocal support for the generally accepted mechanism and are in very good agreement with experimental selectivities. This study shows that E/Z selectivity of non- and semi-stabilized ylides cannot be fully understood without considering the energy of the elimination TS. The influence of ylide stabilization and the nature of phosphorus substituents on reversibility of oxaphosphetane formation is clarified. Unexpectedly, the puckering ability of addition TSs is shown not to depend on ylide stabilization, but the geometry of the TS is decided by an interplay of 1,2; 1,3; and C-H...O interactions in the case of non- and semi stabilized ylides, whereas a dipole-dipole interaction governs the addition TS structures for stabilized ylides. The well-known influence of ylide stabilization on selectivity of PPh(3) derivatives is explained as follows: in non- and semi stabilized ylides reactions, cis and trans addition TSs have, respectively, puckered and planar geometries, and selectivity is governed by an interplay of 1,2 and 1,3 interactions. For stabilized ylides, the high E selectivity is due to a strong dipole-dipole interaction at the addition TS. The influence of the nature of phosphorus substituents on selectivity is also detailed, the different behavior of (MeO)(3)PCHCO(2)Me ylides being explained by their lower dipole. This novel picture of the factors determining TS structures and selectivity provides a sound basis for the design of new ylides. PMID- 16478196 TI - Oxazolidin-2-one-containing pseudopeptides that fold into beta-bend ribbon spirals. AB - Three sets of oligomers containing the 4-carboxy-5-methyloxazolidin-2-one (Oxd) moiety have been synthesized with the aim of checking whether these molecules are able to fold in ordered structures: A set [Boc-(L-Ala-L-Oxd)(n)-OR], B set [Boc (L-Ala-D-Oxd)(n)-OR], and C set [Boc-(Aib-L-Oxd)(n)-OR] preferential conformations have been analyzed with IR absorption, NMR, and CD. We have noticed that in these oligomers three stabilizing effects are active: (i) the rigid Oxd CO-N(CH<)-CO- moiety, which always tend to assume a trans conformation; (ii) the formation of Oxd C=O...H-(alpha)C intramolecolar H-bonds; (iii) the alternate formation of 1 <-- 4 intramolecular C=O...H-N H-bonds. Through the analysis of the experimental data, we could demonstrate that only the oligomers of the B set are able to meet all three requirements listed above. By a deeper insight into the CD spectra, we gathered that the secondary structure adopted by the B set oligomers is a beta-bend ribbon spiral, which is a subtype of the 3(10)-helix. PMID- 16478197 TI - Pattern recognition of proteins based on an array of functionalized porphyrins. AB - A practical protein-detecting array is desirable for its potential application in proteomics, medical diagnostics, and pathogen detection. Here, we report a novel protein-detecting array based on porphyrins containing peripheral amino acids as protein surface receptors. The array of porphyrin receptors showed a unique pattern of fluorescence change upon interaction with certain protein samples. Both metal and nonmetal-containing proteins and mixtures of proteins gave distinct patterns, allowing their unambiguous identification. The composite pattern for each sample was subjected to principal component analysis (PCA) to generate a clustering map for more practical visualization. Increasing the number of porphyrin receptors from eight to sixteen gave improved resolution, suggesting that this array is expandable to give satisfactory resolution for any given sample system by carefully maximizing the chemical diversity of the receptors. PMID- 16478198 TI - Ultrafast cooling of photoexcited electrons in gold nanoparticle-thiolated DNA conjugates involves the dissociation of the gold-thiol bond. AB - Using UV-visible extinction spectroscopy and femtosecond pump-probe transient absorption spectroscopy, we have studied the effect of femtosecond laser heating on gold nanoparticles attached to DNA ligands via thiol groups. It is found that femtosecond pulse excitation of the DNA-modified nanoparticles at a wavelength of 400 nm leads to desorption of the thiolated DNA strands from the nanoparticle surface by the dissociation of the gold-sulfur bond. The laser-initiated gold sulfur bond-breaking process is a new pathway for nonradiative relaxation of the optically excited electrons within the DNA-modified gold nanoparticles, as manifested by a faster decay rate of the excited electronic distribution at progressively higher laser pulse energies. The experimental results favor a bond dissociation mechanism involving the coupling between the photoexcited electrons of the nanoparticles and the gold-sulfur bond vibrations over one involving the conventional phonon-phonon thermal heating processes. The latter processes have been observed previously by our group to be effective in the selective photothermal destruction of cancer cells bound to anti-epidermal growth factor receptor-conjugated gold nanoparticles. PMID- 16478199 TI - Mechanism of the gold(I)-catalyzed Rautenstrauch rearrangement: a center-to-helix to-center chirality transfer. AB - The mechanism of the stereospecific gold(I)-catalyzed Rautenstrauch rearrangement of (E)-1-ethynyl-2-methyl-but-2-en-yl acetate to 3,4-dimethyl-cyclopent-2-enone has been computationally addressed using DFT (B3LYP/6-31G, SDD for Au). Our results indicate that the bond formation event follows the Au(I)-induced acetyl transfer to the vicinal alkyne and that it is the helicity of the pentadienyl cation intermediate which keeps memory of the chiral information. The fidelity of the center-to-helix-to-center chirality transfer requires that the rates of helix interconversion and pivaloyl rotation are slower than the cyclization, as calculations predict. PMID- 16478200 TI - Demonstration of the ring conformation in polyproline by the Raman optical activity. AB - Raman and Raman optical activity (ROA) spectra of poly-L-proline were recorded in a wide frequency range and analyzed with respect to the proline side chain conformation. The analysis was based on comparison to ab initio simulations of spectral band positions and intensities. The presence of two conformer states of the five-member ring was found, approximately equally populated in the polypeptide. Additionally, Raman and ROA spectral shapes indicated that the peptide adopts the polyproline II helical conformation, in both aqueous and TFE solutions. The helix, however, is perturbed by fluctuations, which affects the vibrational coupling among amino acid residues and broadens the ROA bands. Contributions of the side and main peptide chains to the polyproline ROA intensities have comparable magnitudes. Thus understanding of the origins of both signals is important for determination of the peptide structure by ROA. PMID- 16478201 TI - Tetrathiafulvalene-calix[4]pyrroles: synthesis, anion binding, and electrochemical properties. AB - The syntheses of monotetrathiafulvalene-calix[4]pyrrole 5 and bistetrathiafulvalene-calix[4]pyrrole 6, prepared from the acid-catalyzed condensation of monopyrrolo[3,4-d]tetrathiafulvalene (MPTTF, 7) with acetone in the presence of tripyrrane 8 and dipyrromethane 9, respectively, are described. Compound 5 and the previously reported tetrathiafulvalene-calix[4]pyrrole 4 both adopt a 1,3-alternative conformation in the solid state, as determined from X-ray crystallographic analysis. The anion binding properties of the tetrathiafulvalene calix[4]pyrroles 5 and 6, as well as those of the parent meso octamethylcalix[4]pyrrole (1), were investigated in acetone using (1)H NMR spectroscopic and isothermal titration calorimetry (ITC) techniques and, within the error limits of the methods, were generally found to give concordant results. On the basis of the results of the ITC studies carried out in 1,2-dichloroethane, increasing the number of tetrathiafulvalene units annulated to the calix[4]pyrrole system serves to enhance the anion binding affinities substantially but at the price of lowered selectivity. Cyclic voltammetry (CV) studies, carried out in 1,2-dichloroethane, provided evidence of an anion dependent electrochemical response with Cl(-) and Br(-) ions. This response was particularly dramatic in the case of the monotetrathiafulvalene-calix[4]pyrrole 5, with a DeltaE(max) of -145 mV being seen after the addition of approximately 1 equiv of Cl(-) ion. PMID- 16478202 TI - Experimental and computational studies on the mechanism of N-heterocycle C-H activation by Rh(I). AB - Evidence is presented for a proposed mechanism of C-H activation of 3-methyl-3,4 dihydroquinazoline (1) by (PCy(3))(2)RhCl. One intermediate (3), a coordination complex of 1 with (PCy(3))(2)RhCl, was identified along the path to the Rh-N heterocyclic carbene product of this reaction (2). Isotopic labeling and reaction rate studies were used to demonstrate that C-H activation takes place intramolecularly on the reaction coordinate between 3 and 2. Computational studies corroborate the proposed mechanism and suggest that the rate-limiting step is oxidative addition of the C-H bond to the metal center. The consequences of this mechanism for coupling reactions of N-heterocycles that occur via Rh catalyzed C-H bond activation are discussed. PMID- 16478203 TI - Di-organoiron mixed valent complexes featuring "(eta2-dppe)(eta5-C5Me5)Fe" endgroups: smooth class-III to class-II transition induced by successive insertion of 1,4-phenylene units in a butadiyne-diyl bridge. AB - The synthesis and study of a new redox family of symmetric dinuclear iron(II/III) complexes featuring "(eta(2)-dppe)(eta(5)-C(5)Me(5))Fe(CC)" endgroups connected by a bis(diethynyl)-4,4'-biphenyl spacer are reported. The solid-state structures were determined (X-rays) for the homovalent Fe(II)/Fe(II) and Fe(III)/Fe(III) parents. In contrast, the mixed valent (MV) complex 5[PF(6)] has a low thermodynamic stability (Kc around 10) and cannot be isolated in a pure form, but was studied in solution. According to the Robin and Day classification, it constitutes a remarkable example of well-behaved weakly coupled class-II organometallic MV compound. The photodriven metal-metal electron-transfer process takes place over ca. 16 A and corresponds to an electronic coupling of ca. 150 cm(-1) with a reorganization energy of ca. 6250 cm(-1) in dichloromethane. A similar investigation was also conducted in the near-IR range for the known and much more stable MV analogue 3[PF(6)] featuring the 1,4-phenyl unit instead of the 4,4'-biphenyl one (K(c) = 2.6 10(4)). The latter also exhibits a localized valency, but presents a very intense intervalence charge-transfer band (IVCT) with a cutoff on the low-energy side. A much stronger electronic coupling is derived (ca. 1700 cm(-1)) from the band shape for this MV complex in the frame of the two-level model. Although slowed, the electron exchange is not disrupted by insertion of an additional para-phenylene moiety into a 1,4-diethynylaryl bridge. Thus, starting from a compound with a butadiyne-diyl spacer, stepwise para phenylene insertions in the bridge produce a smooth Class-III to Class-II transition for the corresponding MV complexes. PMID- 16478204 TI - Directed evolution and substrate specificity profile of homing endonuclease I SceI. AB - The laboratory evolution of enzymes with tailor-made DNA cleavage specificities would represent new tools for manipulating genomes and may enhance our understanding of sequence-specific DNA recognition by nucleases. Below we describe the development and successful application of an efficient in vivo positive and negative selection system that applies evolutionary pressure either to favor the cleavage of a desired target sequence or to disfavor the cleavage of nontarget sequences. We also applied a previously described in vitro selection method to reveal the comprehensive substrate specificity profile of the wild-type I-SceI homing endonuclease. Together these tools were used to successfully evolve mutant I-SceI homing endonucleases with altered DNA cleavage specificities. The most highly evolved enzyme cleaves the target mutant DNA sequence with a selectivity that is comparable to wild-type I-SceI's preference for its cognate substrate. PMID- 16478205 TI - Combined C-H activation/cope rearrangement as a strategic reaction in organic synthesis: total synthesis of (-)-colombiasin a and (-)-elisapterosin B. AB - The total synthesis of (-)-colombiasin A (2) and (-)-elisapterosin B (3) has been achieved. The key step is a C-H functionalization process, the combined C-H activation/Cope rearrangement, between methyl (E)-2-diazo-3-pentenoate and 1 methyl-1,2-dihydronaphthalenes. When the reaction is catalyzed by dirhodium tetrakis((R)-(N-dodecylbenzenesulfonyl)prolinate), Rh(2)(R-DOSP)(4), an enantiomer differentiation step occurs where one enantiomer of the dihydronaphthalene undergoes the combined C-H activation/Cope rearrangement while the other undergoes cyclopropanation. This sequence controls the three key stereocenters in the natural products such that the remainder of the synthesis is feasible using standard chemistry. PMID- 16478206 TI - Addition of cyclopropyl alkynes to a Brook silene: definitive evidence for a biradical intermediate. AB - The addition of three newly developed mechanistic probes, (trans-2 phenylcyclopropyl)ethyne, (trans,trans-2-methoxy-3-phenylcyclopropyl)ethyne, and (trans,trans-2-methoxy-1-methyl-3-phenylcyclopropyl)ethyne, 1a-c, to a Brook silene, 2-tert-butyl-2-trimethylsiloxy-1,1-bis(trimethylsilyl)-1-silene, 10, was examined. When alkyne 1a was added to silene 10 products derived from a formal ene reaction were obtained. When alkynes 1b-c were added to silene 10, in addition to the typical silacyclobutenes, a variety of silacycloheptenes were obtained in which the cyclopropyl ring had clearly opened. Formal ene-addition products were also produced from the addition of 1b to 10. Based on the relative positions of the phenyl and methoxy substituents within the seven-membered ring of the silacycloheptenes and the known behavior of the alkyne probes under both radical and ionic conditions, it was concluded that a biradical intermediate was formed during the addition of alkynes 1b-c to silene 10. In the addition of alkynes 1a-b to silene 10, the ene products are most likely formed by a competitive pericyclic reaction. We also present a straightforward method for the unambiguous determination of the regiochemistry of silacyclobutenes derived from the cycloaddition of terminal alkynes to silenes. PMID- 16478207 TI - Chemical ecology in wheat plant-pest interactions. how the use of modern techniques and a multidisciplinary approach can throw new light on a well-known phenomenon: allelopathy. AB - A new holistic approach on research into allelopathy was launched in the FATEALLCHEM project. The project was financed by the European Commission in the 5th Framework Programme and involved agronomists, biologists, analytical chemists, organic chemists, environmental chemists, ecotoxicologists, and modelers. Benzoxazinones from wheat (Triticum aestivum L.) were the main group of compounds studied in the project. The project showed that future assessments of an extensive use of allelopathic crops must include the development of validated analytical methods, considerations of relevant concentrations, studies on soil transformation, ecotoxicological studies on individual compounds and mixtures, evaluation on human and mammal toxicity, and joint effect studies on weeds, insects, and pathogens. The project results clearly showed the relevance of optimizing the exploitation of cereal benzoxazinones. Crop rotation is a very traditional practice, which was given less importance for decades but is now regaining its importance in agricultural practice as a means of controlling weed seed banks and soilborne diseases and pests. When using cereals as catch crops and green manure, the allelopathic properties of the cereals could now be much more extensively exploited, choosing varieties with optimal production of benzoxazinones and optimizing the time of sowing in relation to the formation of bioactive metabolites. PMID- 16478208 TI - Isolation and synthesis of allelochemicals from gramineae: benzoxazinones and related compounds. AB - Compounds with a (2H)-1,4-benzoxazin-3(4H)-one skeleton have attracted the attention of phytochemistry researchers since 2,4-dihydroxy-(2H)-1,4-benzoxazin 3(4H)-one (DIBOA) and 2,4-dihydroxy-7-methoxy-(2H)-1,4-benzoxazin-3(4H)-one (DIMBOA) were isolated from plants belonging to the Poaceae family. These compounds exhibit interesting biological properties, such as phytotoxic, antimicrobial, antifeedant, antifungal, and insecticidal properties. These chemicals, in addition to a wide variety of related compounds involved in their metabolism, detoxification mechanisms, and degradation on crop soils and other systems, have high interest and in some cases potential agronomic utility. This paper presents a complete review of the methods employed for their synthetic obtention in addition to some of the authors' own contributions to their chemistry. The degradation and phytotoxicity experiments carried out in ongoing research into the potential agronomic utility of these compounds required large amounts of them, which were obtained from natural sources. This paper presents a modified methodology to access DIMBOA from Zea mays cv. Apache and to obtain 2-O beta-D-glucopyranosyl-2,4-dihydroxy-(2H)-1,4-benzoxazin-3(4H)-one (DIBOA-Glc) and DIBOA from Secale cereale L. New synthetic methodologies were employed for the obtention of the lactams 2-hydroxy-(2H)-1,4-benzoxazin-3(4H)-one and 2-hydroxy-7 methoxy-(2H)-1,4-benzoxazin-3(4H)-one and the malonamic acids N-(2 hydroxyphenyl)malonamic acid and N-(2-hydroxy-7-methoxyphenyl)malonamic acid. The aminophenoxazines 2-amino-7-methoxyphenoxazin-3-one and 2-acetamido-7 methoxyphenoxazin-3-one have been synthesized in the authors' laboratory by novel procedures. All of the methodologies employed allowed the desired compounds to be obtained in high yield and in an easy-to-scale manner. PMID- 16478209 TI - Determination of benzoxazinone derivatives in plants by combining pressurized liquid extraction-solid-phase extraction followed by liquid chromatography electrospray mass spectrometry. AB - A new analytical method based on the use of pressurized liquid extraction (PLE) followed by solid-phase extraction with LiChrolut RP C18 cartridges was evaluated for the sample preparation, extraction, and cleanup of eight naturally occurring benzoxazinone derivatives, 2-beta-D-glucopyranosyloxy-4-hydroxy-1,4-benzoxazin-3 one, 2-beta-D-glucopyranosyloxy-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one, 2,4 dihydroxy-1,4-benzoxazin-3-one (DIBOA), 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3 one, 2-hydroxy-1,4-benzoxazin-3-one, 2-hydroxy-7-methoxy-1,4-benzoxazin-3-one, benzoxazolin-2-one, and 6-methoxybenzoxazolin-2-one in plant samples. Afterward, liquid chromatography-electrospray mass spectrometry, using the selected ion monitoring mode and internal standard (2-MeO-DIBOA, indoxyl-beta-D-glucoside, and quercetin-3-O-rutinoside) quantification method was performed. This paper demonstrates the effectiveness of the PLE method, in conjunction with sensitive and specific mass spectrometric detection, for the quantitative recovery of compounds of the benzoxazinone class from plants. The recoveries of the analytes ranged from 66 to 110% with coefficients of variation ranging from 1 to 14%. This method gave detection limits between 1 and 27 microg/g. The method was applied to foliage and roots of three different wheat cultivars, and the analytes were detected in the range of 11-3261 microg/g of dry weight. PMID- 16478210 TI - Benzoxazinoid allelochemicals in wheat: distribution among foliage, roots, and seeds. AB - In this study, the distribution of eight allelochemicals among the foliage, roots, and seeds of different wheat varieties is reported for two different sampling campaigns, corresponding to two consecutive years. The determination of benzoxazinoid derivatives was performed by combining pressurized liquid extraction--solid-phase extraction followed by liquid chromatography- electrospray mass spectrometry. To the authors' knowledge, there are no previous works about the content of allelochemicals in seed tissue of germinated wheat seedlings. Allelochemicals found in seeds were detected at levels similar to those found in foliage and roots. The results showed that the type of metabolites detected depends strongly on the working up procedure of the plant material, as well as of plant growth stage. A general decrease of the total amount of allelochemical content in the plants was observed with plant age. There was a significant difference in the total amount of benzoxazinoid derivatives in the different wheat varieties analyzed. PMID- 16478211 TI - Concentration of benzoxazinoids in roots of field-grown wheat (Triticum aestivum L.) varieties. AB - Benzoxazinones are naturally occurring secondary metabolites of some Gramineae plants, responsible for their resistance to some pathogenic fungi and for their allelopathic action. Six varieties of winter wheat grown in fields under organic or conventional systems and 11 old accessions were tested for two consecutive seasons and three plant development stages for the concentration in their roots of cyclic hydroxamic acids and their degradation products. This is the first report of six benzoxazinones analyzed in plants grown in the field. An analytical technique employing LC-DAD was used for determination. It was shown that 2,4 dihydroxy-7-methoxy-1,4-benzoxazin-3-one, its degradation product 6 methoxybenzoxazolin-2-one, and the lactam 2-hydroxy-7-methoxy-1,4-benzoxazin-2 one were predominant compounds in all tested samples. Their concentrations significantly differed with plant development stage and season, but no significant differences were found between varieties and between plant cultivation systems. The concentrations of 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA) and its degradation product benzoxazolin-2-one (BOA) were much lower, ranging from 60 to 430 mg/kg of dry matter, depending on accession, stage of development, and season. There was no significant difference found between plants grown in different cultivation systems, but there were significant differences between old and new varieties; concentrations of DIBOA and its derivatives were significantly lower in old accessions. It was concluded that the concentrations of DIBOA and BOA, which are precursors of highly fungicidal 2-aminophenol, 2 amino-3H-phenoxazin-3-one, and 2-acetylamino-3H-phenoxazin-3-one, are theoretically high enough to protect plants against some soilborne pathogens. PMID- 16478212 TI - Quantification of benzoxazinone derivatives in wheat (Triticum aestivum) varieties grown under contrasting conditions in Denmark. AB - Three varieties of winter wheat (Triticum aestivum) were grown in both conventional and organic farming systems. The contents of the benzoxazinone derivatives 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), 2-beta-d glucopyranosyloxy-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA-Glc), 6 methoxybenzoxazolin-2-one (MBOA), 2-hydroxy-7-methoxy-1,4-benzoxazin-3-one (HMBOA), benzoxazolin-2-one (BOA), and 2-hydroxy-1,4-benzoxazin-3-one (HBOA) were analyzed at five growth stages (BBCH 9-10, 12, 21, 31, and 53). Major differences were found between the varieties, with Stakado exhibiting the highest contents. In contrast, only minor and erratic differences were found between the two farming systems, suggesting that the inherent differences in the content of benzoxazinone derivatives of the varieties were not significantly affected by the use of pesticides and synthetic fertilizers. The concentration of benzoxazinone derivatives in the foliage was considerably higher at the early growth stages than later in the growing season, with DIMBOA being the most abundant of the benzoxazinone derivatives. An increase in the concentration was observed in early spring compared to late autumn, suggesting that plants synthesized benzoxazinone derivatives at the commencement of growth in early spring. The concentrations in the roots were considerably lower than in the foliage at the early growth stages but remained relatively constant over time, resulting in a higher concentration than in the foliage at the late growth stages. The results are discussed in relation to previous findings that predominantly originate from experiments done under controlled conditions in either growth cabinets or greenhouses. PMID- 16478213 TI - Effect of 6-methoxybenzoxazolin-2-one (MBOA) on the reproduction rate of the grain aphid (Sitobion avenae F.). AB - Partial host-plant resistance could make a substantial contribution to reducing the damage caused by economically important grain aphids and, therefore, to reduced insecticide use. Naturally occurring hydroxamic acids, in particular 2,4 dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), have been shown to be involved in the resistance of cereals to insects. DIMBOA is unstable in aqueous solutions and has been reported to decompose to 6-methoxybenzoxazolin-2-one (MBOA). MBOA was tested on grain aphids living on artificial diet incorporated with MBOA. From 0 to 0.1 mM, the intrinsic rate of increase (rm) increased. From 0.1 to 0.3 it decreased by 73%. The rm was calculated to be 0 at 1.0 mM. Consequently, even low concentrations of MBOA may reduce the aphid multiplication to a level below the economic damage threshold. Therefore, it is possible to breed wheat varieties with a sufficiently high content of DIMBOA to decrease grain aphid populations. PMID- 16478214 TI - Effects of some benzoxazinoids on in vitro growth of Cephalosporium gramineum and other fungi pathogenic to cereals and on Cephalosporium stripe of winter wheat. AB - The benzoxazolinones benzoxazolin-2(3H)-one (BOA) and 6-methoxybenzoxazolin-2(3H) one (MBOA) and selected degradation products of these compounds were examined for their in vitro antifungal activity against Cephalosporium gramineum, Gaeumannomyces graminis var. graminis, and Fusarium culmorum. BOA was also applied to the soil-incorporated inoculum of C. gramineum to test its capability of reducing Cephalosporium stripe disease in winter wheat. MBOA reduced the mycelial growth of G. graminis var. tritici, C. gramineum, and F. culmorum by 50% (EC50) at the concentrations of 77, 134, and 271 microg/mL of corn meal agar, respectively, and the corresponding BOA EC50 values for the fungi were 11, 189, and 456 microg/mL. BOA degradation products 2-amino-3H-phenoxazin-3-one (APO), 2 acetylamino-3H-phenoxazin-3-one (AAPO), and o-aminophenol (o-AP) were much more inhibitory to the growth of C. gramineum and G. graminis var. tritici than the parent compounds. APO, AAPO, and o-AP EC50 values were found to be as low as 0.58, 4.57, and 1.4 microg/mL, respectively, for C. gramineum and 0.78, 2.18, and 0.80 microg/mL for G. graminis var. tritici. These compounds applied at the corresponding concentrations did not significantly affect the mycelial growth of F. culmorum. The treatment of C. gramineum inoculum with a 1% water solution of BOA resulted in a significant reduction infection of winter wheat with C. gramineum as compared to the control with the untreated inoculum,but this treatment was not as effective as the application of a commercial fungicide. PMID- 16478215 TI - Structure-activity relationship (SAR) studies of benzoxazinones, their degradation products, and analogues. Phytotoxicity on problematic weeds Avena fatua L. and Lolium rigidum Gaud. AB - Avena fatua L. (wild oat) and Lolium rigidum Gaud. (rigid ryegrass) are highly problematic weeds affecting a wide variety of cereal crops worldwide. The fact that both of these weeds have developed resistance to several herbicide groups made them optimal candidates as target organisms for ongoing research about the potential application of allelochemicals and analogue compounds as natural herbicide models. Benzoxazinones, a family of natural allelochemicals present in corn, wheat, and rye, including 2,4-dihydroxy-(2H)-1,4-benzoxazin-3(4H)-one and 2,4-dihydroxy-7-methoxy-(2H)-1,4-benzoxazin-3(4H)-one, together with some degradation products, found in crop soils as well as in other systems, and some synthetic analogues of them were tested on wild oat and rigid ryegrass seeds; the results were statistically treated, and some structure-activity relationships, useful in further development of natural herbicide models, were elucidated. The most active compounds were the synthetic benzoxazinone 2-acetoxy-(2H)-1,4 benzoxazin-3(4H)-one and the degradation product 2-aminophenoxazin-3-one, with highly significant inhibition on the development of both weeds. The ecological role of these compounds is discussed by considering both degradability and phytotoxicity. The bioactivity of aminophenoxazines has been correlated by their aqueous solubility-lipophilicity predicted by means of computational methods. PMID- 16478216 TI - Joint action of benzoxazinone derivatives and phenolic acids. AB - The joint action of binary and ternary mixtures of benzoxazinone derivatives and phenolic acids was studied using the additive dose model (ADM) as reference model. The activity of fixed-ratio mixtures of phenolic acids [ferulic acid (FA), p-coumaric acid (CA), vanillic acid (VA), and p-hydroxybenzoic acid (HBA)] and benzoxazinone derivatives [2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), 6-methoxybenzoxazolin-2-one (MBOA), benzoxazolin-2-one (BOA), 2-aminophenol (AP), and N-(2-hydroxyphenyl)acetamide (HPAA)] on Lolium perenne and Myosotis arvensis root growth was assessed in Petri dishes. Root length was recorded 6 days after seeding, and EC(50) and EC(90) values were estimated using nonlinear regression analyses. The benzoxazinone derivatives were found to be more phytotoxic than the phenolic acids, particularly on M. arvensis. Binary mixtures of phenolic acids responded predominantly additively on both plant species. Deviations from additivity were species-specific with antagonistic responses on L. perenne and synergistic responses on M. arvensis. Similarly, binary mixtures of benzoxazinone derivatives also followed the ADM, although synergistic responses were observed for BOA + AP and BOA + HPAA. Binary and ternary mixtures of benzoxazinone derivatives and phenolic acids responded primarily antagonistically; however, a significant synergistic performance was observed with DIMBOA + FA and DIMBOA + VA on L. perenne. These results do not support the assumption that allelopathic effects of wheat can be attributed to synergistic effects of otherwise weakly active allelopathic compounds, and it is suggested that future research be directed toward identifying and studying the effects of other potential allelochemicals including the degradation products of the most abundant wheat allelochemicals. PMID- 16478217 TI - Herbicidal effects of soil-incorporated wheat. AB - The hydroxamic acid 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) and the benzoxazolinones benzoxazolin-2-one (BOA) and 6-methoxybenzoxazolin-2-one (MBOA) have been identified as important allelochemicals in wheat. This study examines the possibility of exploiting the allelopathic properties of wheat as a weed control strategy by cultivating wheat as a precrop and incorporating plant residues into the soil before the next crop is sown. Different wheat varieties were cultivated in field plots during two seasons in both conventional and organic farming systems. Plants were sampled at various growth stages, and their contents of DIMBOA, MBOA, and BOA were determined by chemical analyses. The wheat samples were incorporated into soil, and the effect on germination and growth of 12 different weed species was examined in pot experiments under controlled conditions. In some cases significant effects were obtained, but the results were inconsistent and the effects were not correlated to the content of DIMBOA, MBOA, and BOA in the incorporated wheat plants. ED50 doses of the pure compounds were estimated in dose-response experiments in Petri dishes, and these turned out to be much higher than the predicted maximum concentrations of DIMBOA, MBOA, and BOA in the soil water following incorporation. The study shows that a prerequisite for exploiting the incorporation of wheat residues as a weed control strategy is the development of wheat varieties with an increased content of allelochemicals. PMID- 16478218 TI - Fate of benzoxazinone allelochemicals in soil after incorporation of wheat and rye sprouts. AB - Growing cereals (especially rye), which are incorporated into the soil to increase soil fertility or organic matter content, is a common practice in crop rotation. The additional sanitizing effect of this incorporation has often been appreciated and is said to be due to leaching of benzoxazinones and subsequent formation of benzoxazolinones. In this study wheat (Stakado) and rye (Hacada) sprouts were incorporated into soil in amounts that simulated agricultural practice. By extraction and subsequent LC-MS analysis the disappearance and appearance of benzoxazinones, benzoxazolinones, and phenoxazinones in soil were followed. In the wheat experiments 6-methoxybenzoxazolin-2-one (MBOA) was detected as the main compound. 2-Hydroxy-7-methoxy-1,4-benzoxazin-3-one (HMBOA) and 2-hydroxy-1,4-benzoxazin-3-one (HBOA) were detected as well. No phenoxazinones were detected. For the rye experiment the picture was more complex. In the first 2 days of incubation MBOA and 2,4-dihydroxy-1,4-benzoxazin 3-one (DIBOA) were detected as the main allelochemicals along with HBOA, HMBOA, and benzoxazolin-2-one (BOA), in decreasing order. Later in the incubation period some 2-amino-3H-phenoxazin-3-one (APO) was detected and the amount of HBOA increased considerably and decreased again. The profiling of the benzoxazinone metabolites and their derivates in soil was dynamic and time-dependent. The highest concentrations of most of the compounds were seen at day 1 after incorporation. A maximum concentration was reached at day 4 for a few of the compounds. This study is the first of its kind that shows the dynamic pattern of biologically active benzoxazinone derivates in soil after incorporation of wheat and rye sprouts. Methods for organic synthesis of HBOA and HMBOA were developed as part of the study. PMID- 16478219 TI - Elucidating the transformation pattern of the cereal allelochemical 6-methoxy-2 benzoxazolinone (MBOA) and the trideuteriomethoxy analogue [D3]-MBOA in soil. AB - To deduce the structure of the large array of compounds arising from the transformation pathway of 6-methoxybenzoxazolin-2-one (MBOA), the combination of isotopic substitution and liquid chromatography analysis with mass spectrometry detection was used as a powerful tool. MBOA is formed in soil when the cereal allelochemical 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) is exuded from plant material to soil. Degradation experiments were performed in concentrations of 400 microg of benzoxazolinone/g of soil for MBOA and its isotopomer 6-trideuteriomethoxybenzoxazolin-2-one ([D3]-MBOA). Previously identified metabolites 2-amino-7-methoxyphenoxazin-3-one (AMPO) and 2-acetylamino 7-methoxyphenoxazin-3-one (AAMPO) were detected. Furthermore, several novel compounds were detected and provisionally characterized. The environmental impact of these compounds and their long-range effects are yet to be discovered. This is imperative due to the enhanced interest in exploiting the allelopathic properties of cereals as a means of reducing the use of synthetic pesticides. PMID- 16478220 TI - Transformation of benzoxazinones and derivatives and microbial activity in the test environment of soil ecotoxicological tests on Poecilus cupreus and Folsomia candida. AB - Benzoxazinones, such as 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) and 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA), and benzoxazolinones, such as 6 methoxy-2-benzoxazolinone (MBOA) and 2-benzoxazolinone (BOA), are biologically active secondary metabolites found in cereals. Because these compounds could be exploited as part of a strategy for reducing the use of synthetic pesticides, ecotoxicological tests were performed recently. In this paper, the transformation of the compounds in the test environment of the ecotoxicological tests was studied. DIMBOA was degraded and partly transformed to MBOA during the period of ecotoxicological testing of the compounds. During testing of MBOA on Poecilus cupreus test media the analysis showed that at the initial concentrations of 2 and 10 mg kg(-1) no MBOA was left after 45 days of testing, but the metabolite 2 amino-phenoxazin-3-one (AMPO) was formed. During testing of BOA on both Folsomia candida and Poecilus cupreus the more biologically active compound 2-amino phenoxazin-3-one (APO) was formed. Thus, the ecotoxicological test results on MBOA and BOA were partly due to the microbial transformation of the compounds during the time of testing. PMID- 16478222 TI - Preliminary analysis of toxicity of benzoxazinones and their metabolites for folsomia Candida. AB - The overall objective of this study was to explore the toxicity of benzoxazinone allelochemicals and their metabolites to Folsomia candida (Collembola: Isotomidae) (Willem, 1902). Experimental tests showed transformation products to have more pronounced toxicity than parent compounds. The underlying relationship between the chemical structure and toxicity was then studied using three dimensional QSAR approaches, and results highlighted the role of the steric contribution. PMID- 16478221 TI - Influence of the soil composition on the effects of benzoxazinoid allelochemicals on two soil nontarget organisms. AB - Seven selected benzoxazinoid allelochemicals and synthetic reference compounds were tested for their lethal and sublethal effects in different field soils and standard soil on Folsomia candida and Poecilus cupreus by applying standard laboratory test procedures. The higher microbial activity in the field soils was most probably responsible for the reduced effects of test compounds on F. candida in the majority of all tests, whereas the higher organic carbon content in field soils was likely the reason for the reduced effects of test substances on P. cupreus. PMID- 16478223 TI - Ecotoxicological effects of benzoxazinone allelochemicals and their metabolites on aquatic nontarget organisms. AB - Before natural plant allelochemicals can be exploited as biological pesticides against weeds and for disease control, more than the effect on target organisms needs to be known. This study presents results of aquatic biotests using four organisms, namely, a water flea, a freshwater alga, a soil alga, and a luminescent bacterium. The tested substances were 10 benzoxazinone derivatives, 3 of them known to be wheat allelochemicals, benzoxazolin-2(3H)-one (BOA), 6 methoxybenzoxazolin-2(3H)-one (MBOA), and 2,4-dihydroxy-7-methoxy-2H-1,4 benzoxazin-3-one (DIMBOA), and 7 identified degradation intermediates and metabolites. For comparison, two commercial pesticide formulations (BAS, Betanal) were tested by applying the same set of biotests. The data set produced could be seen as an ecotoxicological evaluation for effects of allelochemicals against nontarget organisms and as a base for further risk assessment. PMID- 16478224 TI - QSAR models for Daphnia magna toxicity prediction of benzoxazinone allelochemicals and their transformation products. AB - The overall objective of this study is the ecotoxicological characterization of the benzoxazinone 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), the benzoxazolinones benzoxazolin-2-one (BOA) and 6-methoxybenzoxazolin-2-one (MBOA), and their transformation products: phenoxazinones 2-acetylamino-7-methoxy-3H phenoxazin-3-one (AAMPO), 2-acetylamino-3H-phenoxazin-3-one (AAPO), 2-amino-7 methoxy-3H-phenoxazin-3-one (AMPO), and 2-amino-3H-phenoxazin-3-one (APO); aminophenol 2-aminophenol AP); acetamide N-(2-hydroxyphenyl)acetamide (HPAA); and malonamic acid amide N-(2-hydroxyphenyl)malonamic acid (HPMA). A comparison between empirical results and theoretical ones using rules-based prediction of toxicity was done, and it can be concluded that only the degradation metabolites exhibited significant ecotoxic effect. Using synthetic pesticides knowledge, several QSAR models were trained with various approaches and descriptors. The models generated exhibited good internal predictive ability (R(cv)2 > 0.6) and were used to predict the toxicity of the natural compounds studied. PMID- 16478225 TI - Aroma production by tissue cultures. AB - Although plant tissue cultures have been in use for the past hundred years, adapting them for the production of aroma compounds started only in the 1970s. The use of tissue cultures in aroma production has its advantages, because plant cells, unlike whole plants, are not limited to geographic locations or the seasons. Cell mass can be doubled relatively rapidly and can be induced for the production of compounds in a coordinated manner. Compounds can be isolated from cells or the medium with relative ease. Therefore, it would seem to be ideal to use plant cell cultures for the production of aroma compounds. Cell cultures, however, also have some problems. The production of aroma compounds or their precursors is in relatively low amounts, and thus this production method is expensive. Additional expenses are the cost of the medium and the purification of the compounds for food use. Also, cell cultures can only be used effectively in systems for which the biochemical pathway of the aroma compounds is known. In this paper the results of experiments for the use of tissue cultures in the production of vanilla, raspberry, strawberry garlic, and onion aromas is discussed. PMID- 16478226 TI - Qualitative and quantitative polymerase chain reaction analysis for genetically modified maize MON863. AB - Qualitative and quantitative analytical methods were developed for the new event of genetically modified (GM) maize, MON863. One specific primer pair was designed for the qualitative polymerase chain reaction (PCR) method. The specificity and sensitivity of the designed primers were confirmed. PCR was performed on genomic DNAs extracted from MON863, other GM events, and cereal crops. Single PCR product was obtained from MON863 by the designed primer pair. Eight test samples including GM maize MON863 were prepared at 0.01 approximately 10% levels and analyzed by PCR. Limit of detection of the method was 0.01% for GM maize MON863. On the other hand, another specific primer pair and probe were also designed for quantitative method using a real-time polymerase chain reaction. As a reference molecule, a plasmid was constructed from a taxon-specific DNA sequence for maize, a universal sequence for a cauliflower mosaic virus (CaMV) 35S promoter used in most genetically modified organisms, and a construct-specific DNA sequence for the MON863 event. Six test samples of 0.1, 0.5, 1.0, 3.0, 5.0 and 10.0% of GM maize MON863 were quantitated for the validation of this method. At the 3.0% level, the bias (mean vs true value) for MON863 was 3.0%, and its relative standard deviation was 5.5%. Limit of quantitation of the method was 0.5%. These results show that the developed PCR methods can be used to qualitatively and quantitatively detect GM maize MON863. PMID- 16478227 TI - Quantitative analysis and detection of adulteration in crab meat using visible and near-infrared spectroscopy. AB - Visible and near-infrared spectroscopy (VIS/NIR) has been used to detect economic adulteration of crab meat samples. Atlantic blue and blue swimmer crab meat samples were adulterated with surimi-based imitation crab meat in 10% increments. Waveform evaluation revealed that the main features seen in the spectral data arise from water absorptions with a decrease in sample absorbance with increasing adulteration level. Prediction and quantitative analysis was done using raw data, a 15-point smoothing average, a first derivative, a second derivative, and 150 wavelength spectral data gathered from a correlogram. Regression analysis included partial least squares (PLS) and principal component analysis (PCR). Both models were able to perform similarly in predicting crab meat adulteration. The best model for both PLS and PCR used the first derivative spectral data gathered from the correlogram, with a standard error of prediction (SEP) of 0.252 and 0.244, respectively. The results suggest that VIS/NIR technology can be successfully used to detect adulteration in crab meat samples adulterated with surimi-based imitation crab meat. PMID- 16478228 TI - Ion chromatographic determination of perchlorate in foods by on-line enrichment and suppressed conductivity detection. AB - Systemic uptake of perchlorate anion, a rocket fuel component and potential thyroid function disruptor, by leafy vegetables and other crops grown in contaminated waters is a public health concern. A column-switching anion-exchange chromatographic method with suppressed conductivity detection, described in this paper, achieved a 3-6 microg/kg method limit of quantitation in analysis of the wet weight edible portion of cantaloupe, carrots, lettuce, and spinach samples with field-incurred perchlorate. A test portion was blended with dilute nitric acid, and the extract was filtered under vacuum. A portion of the measured filtrate was acidified to pH approximately 2 by addition of cation-exchange resin, 4 mL was passed through a graphitized carbon cleanup column, and an aliquot of a collected fraction was pushed through a short precolumn for anion extraction, enrichment, and injection onto the analytical column. Statistical comparison with determination by tandem mass spectrometry-ion chromatography analysis of untreated filtrate revealed that the difference between means was not significant at the 95% confidence level (P value > or = 0.12) for crops tested. In addition, the method was applied to cooked vegetables processed as baby food. PMID- 16478229 TI - PCR-RFLP authentication of meats from red deer (Cervus elaphus), fallow deer (Dama dama), roe deer (Capreolus capreolus), cattle (Bos taurus), sheep (Ovis aries), and goat (Capra hircus). AB - PCR-RFLP analysis has been applied to the identification of meats from red deer (Cervus elaphus), fallow deer (Dama dama), roe deer (Capreolus capreolus), cattle (Bos taurus), sheep (Ovis aries), and goat (Capra hircus). PCR amplification was carried out using a set of primers flanking a conserved region of approximately 712 base pairs from the mitochondrial 12S rRNA gene. Restriction site analysis based on sequence data from this DNA fragment permitted the selection of MseI, MboII, BslI, and ApoI endonucleases for species identification. The restriction profiles obtained when amplicons were digested with the chosen enzymes allowed the unequivocal identification of all domestic and game meat species analyzed in the present work. PMID- 16478230 TI - Modified 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (abts) method to measure antioxidant capacity of Selected small fruits and comparison to ferric reducing antioxidant power (FRAP) and 2,2'-diphenyl-1-picrylhydrazyl (DPPH) methods. AB - The measurement of antioxidant capacity in fruits differs from that of other biological samples due to their low pH and very low lipophilic antioxidant capacity. In this report, we present a modified 2,2-azino-bis-3 ethylbenzothiazoline-6-sulfonic acid (ABTS) method for fruits and compare its performance with the other commonly used antioxidant methods of 2,2'-diphenyl-1 picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP). The antioxidant capacity and reaction kinetics of four phenolic compounds, two antioxidant standards, and five fruits were also investigated. The modified ABTS method prepared at a pH of 4.5 with sodium acetate buffer is highly stable and easily applied to fruit samples as compared to the standard (pH 7.4) version. The measured antioxidant capacity of samples varied with the assay method used, pH, and time of reaction. Traditional antioxidant standards (trolox, ascorbic acid) displayed stable, simple reaction kinetics, which allowed end point analysis with all of assays. Of the phenolic compounds examined, chlorogenic and caffeic acids exhibited the most complex reaction kinetics and reaction rates that precluded end point analysis while gallic acid and quercetin reached stable end points. All fruit extracts exhibited complex and varied kinetics and required long reaction times to approach an end point. Because the antioxidant capacity of fruit extracts is a function of the array of individual antioxidants present, accurate comparisons among fruit samples require that reaction times be standardized and of sufficient length to reach steady state conditions and that more than one assay be used to describe the total antioxidant activity of fruit samples. PMID- 16478231 TI - Real-time polymerase chain reaction (PCR) quantitative detection of Brassica napus using a locked nucleic acid TaqMan probe. AB - Several countries have introduced mandatory labeling requirements on foods derived from genetically modified organisms. Real-time quantitative Polymerase Chain Reaction (PCR) has quickly become the method of choice in support of these regulations and requires the development of separate PCR assays targeting the transgenic sequence as well as a specific endogenous gene sequence. To develop a Brassica napus-specific PCR assay, partial sequences of the acetyl-CoA carboxylase BnACCg8 gene from B. napus and the closely related Brassica rapa were determined and compared, and a region of unique nucleotide sequence was identified. Universal amplification primers were designed to either side of this region, and a locked nucleic acid TaqMan probe was designed to the B. napus specific sequence. Evaluation of this primer/probe combination indicated a high level of specificity to B. napus: no amplification signal was observed with any other species tested, including five closely related Brassica species. The method was assayed with 14 different B. napus cultivars, and comparable amplification curves were consistently obtained for all. The assay was highly sensitive, with a limit of detection between 1 and 10 haploid copies. Practically, the method was demonstrated to be effective for the detection of processed food samples and for the quantification of Roundup Ready canola content in mixed samples. PMID- 16478232 TI - Morphological and molecular characterization of different Echinochloa spp. and Oryza sativa populations. AB - Echinochloa P. Beauv. is an important genus because many of its species are weeds infesting most paddy fields, which can reduce the rice grain production by up to 80%. A controversy exists about the taxonomy of the genus due to the high level of morphological variations found in these species. Cyhalofop-butyl, an aryloxyphenoxy-propionate herbicide, is used to control Echinochloa spp. in paddy fields, although differences in susceptibility were found between different Echinochloa species. E. colona was highly susceptible [ED50= 34 g of active ingredient (ai) ha(-1)]; very similar results were obtained with the remaining species. By contrast, E. oryzicola (170 g of ai ha(-1)) was less sensitive, with the herbicide symptoms appearing later. Because of this differential susceptibility, morphological and molecular studies were carried out. A morphological study, using 21 characters both quantitative and qualitative of spikelets and seedlings, was capable of clearly distinguishing closely related E. crus-galli plants (two populations), E. muricata and E. crus-pavonis, and E. oryzicola, E. utilis, and E. colona species. The resolution of Echinochloa species at the molecular level, based on RAPD analyses, was fairly consistent with morphological analysis results. Among the 60 primers screened, 21 primers exhibited polymorphic bands and produced a total of 136 RAPD markers. Of all the amplified fragments, 90 were found to be polymorphic. E. oryzicola and E. colona were clearly separated, and the RAPD analyses showed that both E. crus-galli populations were 100% related and 51% related to E. utilis, whereas E. crus pavonis and E. muricata (73% similarity) appeared as being clearly separated from this group. PMID- 16478233 TI - Monolithic supports for the characterization of commercial maize products based on their chromatographic profile. Application of experimental design and classification techniques. AB - Modern analytical techniques based on the use of RP-HPLC with monolithic stationary phases and the application of experimental design and classification tools have been applied to the analysis of maize proteins. Solubilization conditions of maize proteins and separation conditions (temperature, detection wavelength, type and concentration of ion-pairing agent, and gradient) were optimized. The elution gradient was optimized by the application of experimental design techniques. The optimized method consisted of a linear binary gradient of water/acetonitrile/0.1% trifluoroacetic acid in three steps at a flow rate of 3 mL/min with a column temperature of 35 degrees C and UV detection at 280 nm. The developed method enabled the separation of maize proteins in an analysis time close to 8 min. Moreover, this is the first time that commercial maize products have been characterized by the use of multivariate classification techniques. PMID- 16478234 TI - Evaluation of liquid chromatographic behavior of cephalosporin antibiotics using restricted access medium columns for on-line sample cleanup of bovine milk. AB - Microsample deproteinization of bovine milk was carried out on-line using a series of restricted access medium (RAM) bovine serum albumin (BSA) columns: C8, C18, phenyl, and cyano. The four different columns prepared showed a high percentage of protein exclusion using water as the mobile phase and provided an appropriate retention profile for a series of five cephalosporin antibiotics (cefoperazone, cephacetril, cephalexin, cephapirin, and ceftiofur). Chromatographic conditions such as washing time, buffer pH, and type and percentage of organic modifier were fully evaluated with respect to the protein elution profile and retention of the antibiotic by RAM column. One of these columns was chosen to develop and validate a method for the determination of cefoperazone in bovine milk. The system used in this work was composed of a RAM BSA phenyl column coupled to a C18 analytical column. The standard curve was linear over the range 0.100-2.50 microg/mL. The limits of quantification and detection were 0.100 and 0.050 microg/mL, respectively. The developed method showed high intermediate precision (CV of 2.37-2.63%) and accuracy (90.7-94.3%) with adequate sensitivity for drug monitoring in bovine milk samples. PMID- 16478235 TI - Analytical study of trichlorfon residues in Kaki fruit and cauliflower samples by liquid chromatography-electrospray tandem mass spectrometry. AB - A detailed analytical study on trichlorfon residues in selected vegetables samples has been carried out, focused on the reliable quantification and confirmation of this compound, and on stability of residues under storage. As a consequence, a rapid and sensitive LC-ESI-MS/MS method has been developed for the determination of residues of this insecticide in kaki fruit (flesh and peel) and cauliflower samples. Extraction was performed with acetonitrile using a high speed blender. After 4-fold dilution of the extract with water, 20 microL was directly injected in the LC-ESI-MS/MS system (triple quadrupole), using matrix matched standards calibration for quantification. Under optimized MS/MS conditions, limit of detections between 0.006 and 0.013 mg/kg were reached, and a limit of quantification of 0.05 mg/kg was established, with a runtime of only 15 min. Recoveries from spiked blank samples at 0.05 and 0.5 mg/kg were in the range 83-101% with relative standard deviations lower than 10%. The method was applied to treated and untreated samples collected from field residues trials, using quality control samples analysis for the evaluation of the method. Despite the acquisition of two MS/MS transitions in selected reaction monitoring mode, the analysis of treated samples revealed the presence of a chromatographic peak close to the analyte that corresponded to a trichlorfon isobaric compound that shared the same MS/MS transitions. This unusual situation in LC-MS/MS-based procedures required the application of an efficient chromatographic separation to avoid this interference. All experiments have been made in compliance with the principles of Good Laboratory Practices (GLP) and following the European SANCO guidelines for pesticides residue analysis (PRA). PMID- 16478236 TI - Separation of Delta5- and Delta7-phytosterols by adsorption chromatography and semipreparative reversed phase high-performance liquid chromatography for quantitative analysis of phytosterols in foods. AB - A method for the separation, isolation, and identification of phytosterols was developed. A commercial phytosterols mixture, Generol 95S, was fractionated first by adsorption silica gel column chromatography and then separated by means of a semipreparative reverse phase high-performance liquid chromatography fitted with a Polaris C8-A column (250 mm x 10 mm i.d., 5 microm) using isocratic acetonitrile:2-propanol:water (2:1:1, v/v/v) as the mobile phase. Milligram scales of six individual phytosterols, including citrostadienol, campesterol, beta-sitosterol, Delta7-avenasterol, Delta7-campesterol, and Delta7-sitosterol, were obtained. Purities of these isolated sterols were 85-98%. Relative response factors (RRF) of these phytosterols were calculated against cholestanol as an authentic commercial standard. These RRF values were used to quantify by gas chromatography-mass spectrometry (GC-MS) the phytosterols content in a reference material, oils, and chocolates. PMID- 16478237 TI - Counteraction of adriamycin-induced oxidative damage in rat heart by selenium dietary supplementation. AB - Many reports indicate that dietary selenium, potentially increasing the activity of glutathione peroxidase, could offer protection against free-radical-induced damage. The effects of diets moderately enriched in selenium, as sodium selenite or as a lyophilized selenium-rich food, were studied in rats. Adriamycin, an anticancer drug causing a free-radical-mediated cardiotoxicity, was administered intraperitoneally to some rats. The onset of an oxidative damage was indicated by the increase in the plasma level of reactive oxygen metabolites coupled to a decrease in the total antioxidant activity but without modification of glutathione peroxidase activity, which were observed in all rats, independent of the dietary treatment. On the contrary, in the heart, selenium supplementation caused an increase in the total antioxidant activity, glutathione concentration, and glutathione peroxidase and catalase activities leading to a decreased generation of reactive oxygen metabolites. These results clearly indicate that a moderate Se dietary supplementation counteracts adriamycin-induced cardiotoxicity by preservation of endogenous antioxidants. PMID- 16478238 TI - Isolation of an in vitro and ex vivo antiradical melanoidin from roasted barley. AB - The antiradical properties of water-soluble components of both natural and roasted barley were determined in vitro, by means of DPPH* assay and the linoleic acid-beta-carotene system, and ex vivo, in rat liver hepatocyte microsomes against lipid peroxidation induced by CCl4. The results show the occurrence in natural barley of weak antioxidant components. These are able to react against low reactive peroxyl radicals, but offer little protection against stable DPPH radicals deriving from peroxidation in microsomal lipids. Conversely, roasted barley yielded strong antioxidant components that are able to efficiently scavenge free radicals in any system used. The results show that the barley grain roasting process induces the formation of soluble Maillard reaction products with powerful antiradical activity. From roasted barley solution (barley coffee) was isolated a brown high molecular mass melanoidinic component, resistant to acidic hydrolysis, that is responsible for most of the barley coffee antioxidant activity in the biosystem. PMID- 16478239 TI - Pachypodol from Croton ciliatoglanduliferus Ort. as water-splitting enzyme inhibitor on thylakoids. AB - A bioactivity-guided chemical study of aerial parts of Croton ciliatoglanduliferus Ort. led to the isolation for the first time of the flavonoids retusin (5-hydroxy-3,7,3',4'-tetramethoxyflavone) (1) and pachypodol (5,4'-dihydroxy-3,7,3'-trimethoxyflavone) (2) from the n-hexane extract. Compounds 1 and 2 were separated by preparative thin-layer chromatography. Compound 2 was the most active compound on ATP synthesis inhibition. The I50 value was 51 microM. Pachypodol behaves as a Hill reaction inhibitor. It inhibited the uncoupled electron flow on photosystem II partial reaction from water to dichlorophenol indophenol (DCPIP) and from water to sodium silicomolybdate. However, the uncoupled partial reaction from diphenylcarbazide to DCPIP and the uncoupled photosystem I from DCPIPred to MV were not inhibited by 2. These results were corroborated by fluorescence decay data. Therefore, pachypodol inhibits the water-splitting enzyme activity. Compound 1 with a 4' methoxy group was a weak inhibitor, indicating that the 4' free -OH group is important for strong inhibition. PMID- 16478240 TI - Acai (Euterpe oleracea Mart.) polyphenolics in their glycoside and aglycone forms induce apoptosis of HL-60 leukemia cells. AB - The effects of acai polyphenolics on the antiproliferation and induction of apoptosis in HL-60 human leukemia cells were investigated. Interactions between anthocyanins and non-anthocyanin-polyphenolics in both their glycosidic and their aglycone forms were also investigated to determine additive or nonadditive responses. Polyphenolic fractions at 0.17-10.7 microM were found to reduce cell proliferation from 56 to 86% likely due to caspase-3 activation (apoptosis). Anthocyanin and polyphenolic fractions were nonadditive in their contribution to the cell antiproliferation activity. At equimolar concentrations, the glycosidic forms of phenolic acids and flavonoids induced a higher magnitude of change in cell parameters (proliferation and apoptosis) than their respective aglycone forms, while the opposite trend was observed for anthocyanin aglycones. This study demonstrated that acai offers a rich source of bioactive polyphenolics and confirmed the importance of investigating whole food systems when evaluating the potential health benefits of individual phytochemical compounds. PMID- 16478241 TI - Synthesis, structure analyses, and characterization of novel epigallocatechin gallate (EGCG) glycosides using the glucansucrase from Leuconostoc mesenteroides B-1299CB. AB - In this study, three epigallocatechin gallate glycosides were synthesized by the acceptor reaction of a glucansucrase produced by Leuconostoc mesenteroides B 1299CB with epigallocatechin gallate (EGCG) and sucrose. Each of these glycosides was then purified, and the structures were assigned as follows: epigallocatechin gallate 7-O-alpha-D-glucopyranoside (EGCG-G1); epigallocatechin gallate 4'-O alpha-D-glucopyranoside (EGCG-G1'); and epigallocatechin gallate 7,4'-O-alpha-D glucopyranoside (EGCG-G2). One of these compounds (EGCG-G1) was a novel compound. The EGCG glycosides exhibited similar or slower antioxidant effects, depending on their structures (EGCG > or = EGCG-G1 > EGCG-G1' > EGCG-G2), and also manifested a higher degree of browning resistance than was previously noted in EGCG. Also, EGCG-G1, EGCG-G1', and EGCG-G2 were 49, 55, and 114 times as water soluble, respectively, as EGCG. PMID- 16478242 TI - Prolyl endopeptidase inhibitory activity of unsaturated fatty acids. AB - Prolyl endopeptidase (PEP, EC 3.4.21.26) is widely distributed in various organs, particularly in the brains of amnestic patients. Evaluation of PEP levels in postmortem brains of Alzheimer's disease patients revealed significant increases in PEP activity, suggesting that a specific PEP inhibitor can be a good candidate for an antiamnestic drug. In this study, mono- and polyunsaturated fatty acids were investigated to determine their role as PEP inhibitors. Oleic, linoleic, and arachidonic acids, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) showed PEP inhibitory activities (IC50 values of 23.6 +/- 0.4, 43.8 +/- 1.8, 53.4 +/- 1.2, 99.4 +/- 1.2, and 46.2 +/- 1.0 microM, respectively), indicating that they were effective PEP inhibitors, with inhibition constant (Ki) values of 26.7 +/- 0.3, 51.0 +/- 0.7, 91.3 +/- 3.1, 247.5 +/- 2.6, and 89.0 +/- 2.3 microM, respectively. Oleic acid showed the highest PEP inhibitory activity. Dixon plots of PEP inhibition showed oleic, linoleic, and arachidonic acids, EPA, and DHA are noncompetitive inhibitors; despite higher IC50 values of these unsaturated fatty acids than strong natural inhibitors, they may have potential use in preventing memory loss. PMID- 16478243 TI - Antioxidant activity and inhibition of alpha-glucosidase by trans-resveratrol, piceid, and a novel trans-stilbene from the roots of Israeli Rumex bucephalophorus L. AB - The roots of Rumex bucephalophorus, collected in Israel, were analyzed for trans stilbenes. Two stilbene-O-glycosyl derivatives were identified, in addition to 3,5,4'-trihydroxystilbene (1) (resveratrol). The stilbene-O-glycosyl derivatives were 5,4'-dihydroxystilbene-3-O-beta-d-glucopyranoside (2) (piceid) and the new 5,4'-dihydroxystilbene-3-O-alpha-arabinopyranoside (3), which is being named rumexoid. The structure of rumexoid was elucidated by using spectroscopic data. The antioxidant capacities of stilbenoids 1-3 were determined and expressed as trolox equivalent antioxidant capacity (TEAC). TEAC value for trans-resveratrol was highest (2.7) and for rumexoid lowest (1.5). In vitro, trans-resveratrol and rumexoid demonstrated a potent inhibitory effect on alpha-glucosidase activity (IC50 < 0.1 and < 0.5 mM, respectively). The commercial antidiabetic agent acarbose was shown to inhibit only 35% of the enzyme activity at 0.5 mM. The addition of piceid to the reaction mixture did not inhibit alpha-glucosidase in vitro in the range of concentrations used. These findings extend the range of reported beneficial effects of stilbene derivatives, and demonstrate the multifaceted activities that dietary polyphenols may exert in the intestine, where their concentrations are highest in the body. PMID- 16478244 TI - Antioxidant levels and inhibition of cancer cell proliferation in vitro by extracts from organically and conventionally cultivated strawberries. AB - The effects of extracts from five cultivars of strawberries on the proliferation of colon cancer cells HT29 and breast cancer cells MCF-7 were investigated, and possible correlations with the levels of several antioxidants were analyzed. In addition, the effects of organic cultivation compared to conventional cultivation on the content of antioxidants in the strawberries and strawberry extracts on the cancer cell proliferation were investigated. The ratio of ascorbate to dehydroascorbate was significantly higher in the organically cultivated strawberries. The strawberry extracts decreased the proliferation of both HT29 cells and MCF-7 cells in a dose-dependent way. The inhibitory effect for the highest concentration of the extracts was in the range of 41-63% (average 53%) inhibition compared to controls for the HT29 cells and 26-56% (average 43%) for MCF-7 cells. The extracts from organically grown strawberries had a higher antiproliferative activity for both cell types at the highest concentration than the conventionally grown, and this might indicate a higher content of secondary metabolites with anticarcinogenic properties in the organically grown strawberries. For HT29 cells, there was a negative correlation at the highest extract concentration between the content of ascorbate or vitamin C and cancer cell proliferation, whereas for MCF-7 cells, a high ratio of ascorbate to dehydroascorbate correlated with a higher inhibition of cell proliferation at the second highest concentration. The significance of the effect of ascorbate on cancer cell proliferation might lie in a synergistic action with other compounds. PMID- 16478245 TI - Importance of insoluble-bound phenolics to antioxidant properties of wheat. AB - Two commercial samples of soft (70% Canadian Eastern soft red spring and 30% Canadian Eastern soft white winter) and hard (90% Canadian western hard red spring and 10% Canadian Eastern hard red winter) wheats were used to obtain different milling fractions. Phenolics extracted belonged to free, soluble esters and insoluble-bound fractions. Soluble esters of phenolics and insoluble-bound phenolics were extracted into diethyl ether after alkaline hydrolysis of samples. The content of phenolics was determined using Folin-Ciocalteu's reagent and expressed as ferulic acid equivalents (FAE). The antioxidant activity of phenolic fractions was evaluated using Trolox equivalent antioxidant capacity, 2,2 diphenyl-1-picrylhydrazyl radical scavenging, reducing power, oxygen radical absorbance capacity, inhibition of oxidation of human low-density lipoprotein cholesterol and DNA, Rancimat, inhibition of photochemilumenescence, and iron(II) chelation activity. The bound phenolic content in the bran fraction was 11.3 +/- 0.13 and 12.2 +/- 0.15 mg FAE/g defatted material for hard and soft wheats, respectively. The corresponding values for flour were 0.33 +/- 0.01 and 0.46 +/- 0.02 mg FAE/g defatted sample. The bound phenolic content of hard and soft whole wheats was 2.1 (+/-0.004 or +/-0.005) mg FAE/g defatted material. The free phenolic content ranged from 0.14 +/- 0.004 to 0.98 +/- 0.05 mg FAE/g defatted milling fractions of hard and soft wheats examined. The contribution of bound phenolics to the total phenolic content was significantly higher than that of free and esterified fractions. In wheat, phenolic compounds were concentrated mainly in the bran tissues. In the numerous in vitro antioxidant assays carried out, the bound phenolic fraction demonstrated a significantly higher antioxidant capacity than free and esterified phenolics. Thus, inclusion of bound phenolics in studies related to quantification and antioxidant activity evaluation of grains and cereals is essential. PMID- 16478246 TI - Genotype and environmental variation in phenolic content, phenolic acid composition, and antioxidant activity of hard spring wheat. AB - The health-promoting effects of whole-grain wheat likely derive from phenolic compounds and other antioxidants that also make wheat a potential source of functional food ingredients. The objective of this study was to determine the effects of genotype and growing environment on the phenolic contents and antioxidant activities of alcohol-soluble extracts from commercial wheat cultivars. Total phenolic contents (TPCs), antioxidant activities (AOAs), and concentrations of six phenolic acids were measured in six red- and white-grained hard spring wheat genotypes grown at four diverse locations in Western Canada during the 2003 crop year. There were significant differences among genotypes and environments for TPC, AOA, and concentrations of all the phenolic acids measured. The predominant indicators of antioxidant potential, i.e., TPC, AOA, and ferulic acid (FA) concentration were highly intercorrelated (r > 0.72). For these indices, the Canada Western (CW) Red Spring wheat cultivars Neepawa and AC Elsa had the highest levels, whereas an analogous CW hard white spring wheat cultivar, AC Snowbird, had the lowest levels. Grain color did not appear to be a factor in the expression of antioxidant-related parameters. For both TPC and AOA, as well as for vanillic acid, syringic acid, and ferulic acid, environmental effects were considerably larger than genotype effects. Neither growing temperature nor rainfall from anthesis to maturity appeared to be related to the environmental variation that was observed. Genotype x environment interaction was small for all parameters compared with genotype and location effects and was significant only for TPC. Genotype variation for antioxidant properties indicates that it would be possible to select for these quantitative traits in a breeding program. However, the significant environmental variation observed would delay and/or complicate this process. PMID- 16478247 TI - Polyphenol composition and antioxidant activity of Kei-apple (Dovyalis caffra) juice. AB - The polyphenolic and ascorbate (ASC) components as well as the antioxidant capacity of Kei-apple (Dovyalis caffra) juice were analyzed and compared to three other fruit juices. The Kei-apple juice had significantly the highest total polyphenolic concentrations (1013 mg gallic acid equivalent/L), and solid phase (C(18)) fractionation identified the majority of these polyphenols to be phenolic acids. The Kei-apple juice also had significantly the highest ASC concentrations (658 mg/L), which showed exceptional heat stability with very little conversion to dehydroascorbate (DHA). Antioxidant capacities of both the unfractionated fruit juices and their solid phase-extracted fractions, as determined by oxygen radical absorbance capacity and ferric reducing antioxidant power analyses, correlated well to the polyphenol concentrations. Gas chromatography-mass spectrometry analyses showed caffeic acid as the most abundant polyphenol present (128.7 mg/L) in the Kei-apple juice; it contributed to 63% of the total antioxidant capacity (of all of the individual compounds identified). Other notable polyphenols identified in higher concentrations included p-coumaric acid, p-hydroxyphenylacetic acid, and protocatechuic acid. Our results therefore support the putative high antioxidant value linked to this fruit and better define this potential in terms of the major antioxidants that exist in the Kei apple. PMID- 16478248 TI - Seasonal variation of red clover (Trifolium pratense L., Fabaceae) isoflavones and estrogenic activity. AB - Red clover (Trifolium pratense L., Fabaceae) dietary supplements are currently used to treat menopausal symptoms because of their high content of the mildly estrogenic isoflavones daidzein, genistein, formononetin, and biochanin A. These compounds are estrogenic in vitro and in vivo, but little information exists on the best time to harvest red clover fields to maximize content of the isoflavones and thus make an optimal product. Samples of cultivated red clover above-ground parts and flower heads were collected in parallel over one growing season in northeastern Illinois. Generally, autohydrolytic extracts of above-ground parts contained more isoflavones and had more estrogenic activity in Ishikawa endometrial cells as compared with extracts of flower heads. Daidzein and genistein contents peaked around June to July, while formononetin and biochanin A contents peaked in early September. Flower head and total above-ground parts extracts exhibited differential estrogenic activity in an Ishikawa (endometrial) cell-based alkaline phosphatase induction assay, whereas nondifferential activity was observed for most extracts tested in an MCF-7 (breast) cell proliferation assay when tested at the same final concentrations. Ishikawa assay results could be mapped onto the extracts' content of individual isoflavones, but MCF-7 results did not show such a pattern. These results suggest that significant metabolism of isoflavones may occur in MCF-7 cells but not in Ishikawa cells; therefore, caution is advised in the choice of bioassay used for the biological standardization of botanical dietary supplements. PMID- 16478249 TI - Properties of aged montmorillonite-wheat gluten composite films. AB - The properties of new and aged glycerol-plasticized vital wheat gluten films containing < or =4.5 wt % natural or quaternary ammonium salt modified montmorillonite clay were investigated. The films were cast from pH 4 or pH 11 ethanol/water solutions. The films, aged for < or =120 days, were characterized by tensile testing, X-ray diffraction, and transmission electron microscopy. In addition, water vapor permeability (11% relative humidity) and the content of volatile components were measured. The large reduction in the water vapor permeability with respect to the pristine polymer suggests that the clay platelets were evenly distributed within the films and oriented preferably with the platelet long axis parallel to the film surface. The film prepared from pH 11 solution containing natural clay was, as revealed by transmission electron microscopy and X-ray diffraction, almost completely exfoliated. This film was consequently also the strongest, the stiffest, and the most brittle and, together with the pH 11 film containing modified clay, it also showed the greatest decrease in water vapor permeability. The large blocking effect of the clay had no effect on the aging kinetics of the films. During aging, the pH 4 and pH 11 film strength and the pH 4 film stiffness increased and the pH 4 film ductility decreased at the same rate with or without clay. This suggests that the aging was not diffusion rate limited, that is, that the loss of volatile components or the migration of glycerol or glycerol/wheat gluten phase separation was not limited by diffusion kinetics. The aging rate seemed to be determined by slow structural changes, possibly involving protein denaturation and aggregation processes. PMID- 16478250 TI - Gene expression profiles of Drosophila melanogaster exposed to an insecticidal extract of Piper nigrum. AB - Black pepper, Piper nigrum L. (Piperaceae), has insecticidal properties and could potentially be utilized as an alternative to synthetic insecticides. Piperine extracted from P. nigrum has a biphasic effect upon cytochrome P450 monooxygenase activity with an initial suppression followed by induction. In this study, an ethyl acetate extract of P. nigrum seeds was tested for insecticidal activity toward adult Musca domestica and Drosophila melanogaster. The effect of this same P. nigrum extract upon differential gene expression in D. melanogaster was investigated using cDNA microarray analysis of 7380 genes. Treatment of D. melanogaster with P. nigrum extract led to a greater than 2-fold upregulation of transcription of the cytochrome P450 phase I metabolism genes Cyp 6a8, Cyp 9b2, and Cyp 12d1 as well as the glutathione-S-transferase phase II metabolism gene Gst-S1. These data suggests a complex effect of P. nigrum upon toxin metabolism. PMID- 16478251 TI - Enzymatic degradation of oligosaccharides in pinto bean flour. AB - The use of dry edible beans is limited due to the presence of flatulence factors, the raffinose oligosaccharides. Our objective was to investigate the process for the removal of oligosaccharides from pinto bean using enzymatic treatment and to compare it to removal by soaking and cooking methods. Crude enzyme preparation was produced by six fungal species on wheat bran- and okara-based substrates with soy tofu whey. The loss of raffinose oligosaccharides after soaking pinto beans for 16 h at the room temperature was 10%, after cooking for 90 min was 52%, and after autoclaving for 30 min was 58%. On the other hand, the treatment using crude alpha-galactosidase (60 U mL(-1)) produced by Aspergillus awamori NRRL 4869 from wheat bran-based substrate with soy tofu whey on pinto bean flour for 2 h completely hydrolyzed raffinose oligosaccharides. These results supported that the enzymatic treatment was the most effective among various processing methods tested for removing the raffinose oligosaccharides, and hence, crude alpha galactosidases from fungi have potential use in the food industry. PMID- 16478252 TI - Modeling bleaching of tomato derivatives at subzero temperatures. AB - This work was addressed to obtain a predictive model of the rate of bleaching in tomato derivatives at subzero temperatures. To this aim, a tomato puree was freeze-dried and equilibrated at increasing solid fractions. The bleaching rate was assessed by measuring tomato color during storage for up to 18 months at temperatures from -30 to 0 degrees C. The temperature dependence of the tomato bleaching rate was neither predictable using the Arrhenius equation nor simply related to tomato physical state. The lack of a clear Arrhenius relation was attributed to the occurrence of temperature-dependent phenomena, such as ice crystallization and oxygen solubility modifications, which strongly changed the local concentration of reactants. A modified Arrhenius equation predicting the tomato-bleaching rate in the entire temperature range was proposed. Tomato concentration, and hence its physical state, affected the temperature dependence of bleaching, modifying apparent activation energy and frequency factor of the modified Arrhenius equation. In light of these considerations, a mathematical model was set up and validated to accurately predict the tomato-bleaching rate on the basis of only its concentration and storage temperature. PMID- 16478253 TI - Distribution profiles of isoflavone isomers in black bean kojis prepared with various filamentous fungi. AB - This study was conducted to compare the transformation of both isoflavone derivatives (aglycones, beta-glucosides, and acetyl and malonyl gluclucosides) and beta-glucosidase activity in kojis fermented with various generally recognized as safe filamentous fungi including Aspergillus awamori, Aspergillus oryzae, Aspergillus sojae, Rhizopus azygosporus, and Rhizopus sp. no. 2. Solid fermentation was performed to prepare the kojis by inoculating the steamed black beans with starter organism and culturing at 30 degrees C for 3 days. Results revealed that fermentation caused a marked increase in the content of aglycone (daidzein, glycitein, and genistein), the bioactive isoflavone, and a significant reduction in the content of beta-glucoside isoflavone (daidzin, glycitin, and genistin), compared with the unfermented steamed black bean. The extent of increased aglycone and reduced beta-glucoside isoflavone content varied with the starter organism used. Among the various black bean kojis prepared, the Rhizopus sp. no. 2 koji showed the highest level of enhancement in aglycone content. In the Rhizopus sp. no. 2 koji, the percentage of aglycone to total isoflavone increased from an initial approximately 2.9 to approximately 58.9% after fermentation. In comparison, the percentages found in kojis prepared with other starter organisms ranged from 18.9 to 38.9% after fermentation. Further preparations of black bean kojis with A. awamori at different cultivation temperatures (25, 30, and 35 degrees C) and various fermentation periods (1-5 days) revealed that koji prepared at 30 degrees C for 4 days showed the highest content of aglycones, with 7.7-, 5.7-, and 4.8-fold increases in the content of daidzein, genistein, and glycitein, respectively. In addition, the increase of aglycone content and the increase of beta-glucosidase activity during the fermentation of this koji showed a similar trend. PMID- 16478254 TI - Effects of roasting on hazelnut lipids. AB - The effect of roasting on some nutritional characteristics of hazelnut lipidic fraction was investigated. Hazelnuts (Corylus avellana L.) were submitted to several different thermal treatments, comprising different temperatures (125-200 degrees C) and times of exposure (5, 15, and 30 min) and analyzed for their moisture and crude fat. Raw and roasted hazelnuts were also analyzed for their compositions in phytosterols and fatty acids (including trans isomers) by GC-FID, triacylglycerols by HPLC-ELSD, and tocopherols and tocotrienols by HPLC DAD/fluorescence spectroscopy. Minor changes occurred in the fatty acid and triacylglycerol compositions. As temperatures and roasting periods increased, generally, a modest increase of oleic and saturated fatty acids and a decrease of linoleic acid, expressed as relative percentages, occurred. Similarly, an increase of triacylglycerols containing oleic acid moieties and a decrease of those containing linoleic acid moieties were found in the roasted samples. Roasting caused a modest decrease of the beneficial phytosterols (maximum 14.4%) and vitamin E homologues (maximum 10.0%) and a negligible increase of the trans fatty acids. PMID- 16478255 TI - Combined enzymatic and high-pressure processing affect cell wall polysaccharides in berries. AB - The effect of high-pressure processing (HPP) on cell wall polysaccharides in berries was investigated. HPP decreased the degree of methyl esterification (DM), probably by activation of pectin methyl esterase (PME), and improved the extractability of pectins. When commercial enzyme mixtures were added to mashed berries, a synergistic effect was observed between treatment with commercial enzymes and HPP. Compared to treatment at atmospheric pressure, pectic polysaccharides were degraded to a larger extent when HPP was used. In contrast, hemicelluloses were hardly affected by the added enzymes when HPP was included, although they were degraded during similar treatment at atmospheric pressure. Additionally, the activity of rhamnose-releasing enzymes present in minor quantities might be enhanced after HPP, resulting in a decrease of rhamnose in the polymeric cell wall material. These results exploring the effect of HPP at representative conditions clearly point out the potential of HPP for polysaccharide modification. PMID- 16478256 TI - Tocopherol and tocotrienol content of hazelnut cultivars grown in portugal. AB - Hazelnuts from 19 cultivars collected during 3 consecutive-year crops, in 2 different geographical localities, for a total of 79 samples, were evaluated for their contents in tocopherols and tocotrienols by normal-phase high-performance liquid chromatography coupled to a series arrangement of a diode array and a fluorescence detector. Seven compounds were identified and quantified. All samples presented alpha-, beta-, gamma-, and delta- tocopherols and beta tocotrienol; alpha- and gamma-tocotrienols were detected in some of the studied samples. alpha-Tocopherol was the major compound in all samples, ranging from 105.9 to 226.8 mg/kg of hazelnut. Considering the generality of the obtained results, an identical qualitative and quantitative pattern was found, which can define a chemical fingerprint that may be helpful in the assessment of identity and quality of hazelnut oils. Statistical analyses were carried out in order to check for differences among cultivars, year crops, and geographical localities. Although some differences were observed when different-year crops and geographical localities were considered, neither of these factors seemed to produce considerable differences in terms of tocopherol and tocotrienol contents. Some minor differences were observed among cultivars. PMID- 16478257 TI - Characterization of volatile compounds of Mezcal, an ethnic alcoholic beverage obtained from Agave salmiana. AB - Commercial mezcals (white, white with worm, rested, rested with worm, and aged) produced from Agave salmiana were analyzed by solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS). Thirty-seven compounds were identified, and nine of them were classified as major compounds of mezcal (MCM). Saturated alcohols, ethyl acetate, ethyl 2-hydroxypropanoate, and acetic acid form the MCM group. Minor compounds of mezcal group include other alcohols, aldehydes, ketones, large chain ethyl esters, organic acids, furans, terpenes, alkenes, and alkynes. Most of the compounds found in mezcals in this study are similar to those present in tequilas and other alcoholic beverages. However, mezcals contain unique compounds such as limonene and pentyl butanoate, which can be used as markers for the authenticity of mezcal produced from A. salmiana. PMID- 16478258 TI - HPLC-DAD/MS characterization of flavonoids and hydroxycinnamic derivatives in turnip tops (Brassica rapa L. Subsp. sylvestris L.). AB - Flavonoids and hydroxycinnamic derivatives of turnip tops (Brassica rapa L. subsp. sylvestris L.) were characterized for the first time in four samples from different origins. Turnip tops exhibit a high polyphenols content (ranging from 107 to 191 mg/100 g, fresh weight) and a good antiradical activity, determined with the DPPH* test. After a liquid-liquid extraction and fractionation procedures, most flavonoids (isorhamnetin, kaempferol, and quercetin glycosides) and hydroxycinnamic derivatives were identified by means of HPLC-DAD/MS techniques. Isorhamnetin glycosides were the main flavonoid derivatives, differing from that found in the vegetables belonging to the Brassica oleracea group. PMID- 16478259 TI - The biological and chemical variability of yacon. AB - This paper focuses on the biological and chemical variability of four yacon (Smallanthus sonchifolius) accessions cultivated under field conditions. Significant variations in tuber shape, weight, content of oligofructans, as well as in leaf isozymes, phenolics, and relative DNA contents were found. Accessions 6 and 88 were the most productive (up to 3.01 and 3.74 kg/plant); accession 48 was the most balanced from the yield aspect in three vegetative periods. A significantly higher content of beta-(2-->1) oligofructans was noted in accessions 48 and 88 as compared to 6 and 60. No difference in sucrose, glucose, and fructose level was observed. Only accession 6 exhibited separate acid phosphatase and esterase isoforms. Accessions 6 and 60 had the highest content of phenolics, and accession 88 had the lowest relative DNA content. Large yacon intraspecific variation may be useful in future detailed research as a good background for breeding, growing, and utilization in industrial processing. PMID- 16478260 TI - The effect of fungicidal treatment on selected quality parameters of barley and malt. AB - Protection of barley grain against contamination by fungi such as Fusarium spp., particularly by those producing mycotoxins, secondary metabolites with adverse health effects, is of principal importance. Fungicides applied immediately after full heading of spring barley is one method of direct protection. In this work, extensive two-year field experiments combined with a detailed chemical laboratory analysis (barley and malt) were performed with the aim to study the effect of previous crops, different fungicides, and other conditions on the selected barley and malt quality parameters (content of beta-glucans, pentosans, oxalic acid, deoxynivalenol, and gushing), while the main task was to follow the effect of the fungicide (used as a treatment to protect against pathogens, mostly Fusarium) on changes of the chemical composition in barley and malt, and gushing. It was found that the relationship between the studied factors and the parameters usually applied to the evaluation of barley and malt quality is quite complex and not straightforward. The responses show typical features of a multifactorial influence with both positive and negative correlations resulting in a decrease or increase in grain quality (concentrations of beta-glucans, pentosans, deoxynivalenol, and other studied parameters). The role of previous crops was also found to be important. The fungicides should be applied at the time of heading but not at the very beginning of this period. PMID- 16478261 TI - Synthesis and structure-activity relationships of 1-phenyl-1H-1,2,3-triazoles as selective insect GABA receptor antagonists. AB - To study the interaction of phenylheterocycles with gamma-aminobutyric acid (GABA) receptors, 4- or 5-alkyl(or phenyl)-1-phenyl-1H-1,2,3-triazoles were synthesized and examined for their ability to inhibit the specific binding of [3H]-4'-ethynyl-4-n-propylbicycloorthobenzoate (EBOB), a noncompetitive antagonist, to the housefly and rat GABA receptors, as well as to the beta3 subunit homo-oligomer of the human GABA receptor investigated as a model receptor. 4-Substituted 1-phenyl-1H-1,2,3-triazoles were found to be more potent competitive inhibitors than the 5-substituted regioisomers in the case of all receptors. The 4-tert-butyl or 4-n-propyl analogue of 1-(2,6-dichloro-4 trifluoromethylphenyl)-1H-1,2,3-triazole exhibited the highest level of inhibition of [3H]EBOB binding to all receptors. Most of the synthesized analogues were more active in terms of the inhibition of EBOB binding to the housefly and human beta3 GABA receptors than to the rat receptor. The 4 cyclohexyl analogue showed the highest (185-fold) housefly versus rat receptor selectivity. A three-dimensional quantitative structure-activity relationship (3D QSAR) analysis demonstrated that both the 4-trifluoromethyl-2,6-dichloro substitution on the phenyl ring and a small, bulky, hydrophobic substituent at the 4-position of the triazole ring played significant roles in conferring high potency in cases involving the housefly and human beta3 receptors. The human beta3 receptor resembled the housefly receptor in terms of their recognition of phenyltriazoles, whereas 3D-QSAR analysis revealed a slight difference between the two receptors in terms of their mechanisms of recognition of the para substituent on the phenyl moiety. Some of the triazoles synthesized here exhibited insecticidal activity, which was correlated with their ability to inhibit [3H]EBOB binding to the housefly receptor. Thus, 1-phenyl-1H-1,2,3 triazoles with the appropriate substituents exert insecticidal activity by selectively acting at the site for noncompetitive antagonism of insect GABA receptors. PMID- 16478262 TI - Influence of soil aging on sorption and bioavailability of simazine. AB - Characterization of pesticide bioavailability, particularly in aged soils, is of continued interest because this information is necessary for environmental risk assessment. However, pesticide bioavailability in aged soils has been characterized by a variety of methods with limited success, due in part to methodological limitations. The objective of this study was to use solvent extraction methods to correlate simazine residue bioavailability in aged soils to simazine mineralization using a simazine-mineralizing bacterium. Soils from Brazil, Hawaii, and the midwestern United States were treated with UL-ring labeled [14C]simazine and incubated for up to 8 weeks. At the end of each incubation period, soils were either incubated further, extracted with 0.01 M CaCl2, or extracted with aqueous methanol (80:20 v/v methanol/water). In a parallel experiment, after each incubation period, soils were inoculated with the bacterium Pseudomonas sp. strain ADP, which is capable of rapidly mineralizing simazine, and 14CO2 was determined. The inoculated soil samples were then extracted with 0.01 N CaCl2 and with aqueous methanol. This allowed for the evaluation of the bioavailability of aged simazine residues, without the contribution of simazine desorption from soil. Results of these studies indicated that simazine sorption to soil increased with aging and that amounts of simazine in aged soils extracted by 0.01 M CaCl2 and aqueous methanol were highly correlated to amounts of simazine mineralized by Pseudomonas sp. strain ADP. Consequently, 0.01 M CaCl2/methanol-extractable simazine in aged soils can be used to estimate bioavailable residues. This technique may be useful in determining the bioavailability of other s-triazine compounds in soils. PMID- 16478263 TI - Chromatographic determination of Fe chelated by ethylenediamine-N-(o hydroxyphenylacetic)-N'-(p-hydroxyphenylacetic) acid in commercial EDDHA/Fe3+ fertilizers. AB - EDDHA/Fe3+ chelates are the most common fertilizers used to solve Fe chlorosis in established crops. Commercial products contain two regioisomers, ethylenediamine N,N'-bis(o-hydroxyphenylacetic) acid (o,o-EDDHA)/Fe3+ and ethylenediamine-N-(o hydroxyphenylacetic)-N'-(p-hydroxyphenylacetic) acid (o,p-EDDHA)/Fe3+. Although several chromatographic methods exist for the determination of Fe3+ chelated by the o,o-EDDHA isomer, no method has been described for the quantification of Fe3+ chelated by o,p-EDDHA. In this work, factors that affect the behavior of o,p EDDHA/Fe3+ in ion pair chromatography are reviewed: pH, ion pair reagent, and organic modifier. The best chromatographic performance was obtained with an aqueous mobile phase at pH 6.0 containing 35% acetonitrile and 5 mM tetrabutylammonium hydroxide under isocratic elution conditions. This method was applied to the quantification of commercial samples. PMID- 16478264 TI - Fe uptake from meso and D,L-racemic Fe(o,o-EDDHA) isomers by strategy I and II plants. AB - One of the most efficient fertilizers to correct Fe deficiency in calcareous soils and waters with high bicarbonate content is based on ferric ethylenediamine N,N'-bis(o-hydroxyphenylacetic) acid [Fe(o,o-EDDHA)]. Fe(o,o-EDDHA) forms two groups of geometric isomers known as meso and D,L-racemic. To determine the Fe uptake from meso and D,L-racemic Fe(o,o-EDDHA), four iron-efficient plants, two plants representative of strategy I (tomato and pepper) and two plants representative of strategy II (wheat and oats), were grown in hydroponic culture. Results indicated that strategy II plants took up iron from both Fe(o,o-EDDHA) isomers equally. However, strategy I plants took mainly the iron associated with the meso form (the lowest stability isomer). PMID- 16478265 TI - Preparation and properties of a double-coated slow-release and water-retention urea fertilizer. AB - A double-coated, slow-release, and water-retention urea fertilizer (DSWU) was prepared by cross-linked poly(acrylic acid)-containing urea (PAAU) (the outer coating), polystyrene (PS) (the inner coating), and urea granule (the core). Elemental analysis results showed that the nitrogen content of the product was 33.6 wt %. The outer coating (PAAU) regulated the nitrogen release rate and protected the inner coating from damage. The slow-release property of the product was investigated in water and in soil. The possible mechanism of nitrogen release was proposed. The influences of PS coating percentage, temperature, water absorbency, and pH on the release of nitrogen were also investigated. It was found that PS coating percentage, temperature, and water absorbency had a significant influence on the release of nitrogen. However, the pH had no effect. The water-retention property of the product was also investigated. The results showed that the product not only had a good slow-release property but also excellent water-retention capacity, which could effectively improve the utilization of fertilizer and water resources. The results of the present work indicated that the DSWU would find good application in agriculture and horticulture, especially in drought-prone areas where the availability of water is insufficient. PMID- 16478266 TI - Thermospectroscopic study of the adsorption mechanism of the hydroxamic siderophore ferrioxamine B by calcium montmorillonite. AB - The behavior of iron-chelating agents in soils is highly affected by interactions with the solid phase. Still this aspect is frequently ignored. In this research the adsorption of the siderophore ferrioxamine B by Ca-montmorillonite, as a free ligand (desferrioxamine B, DFOB) and as a complex with Fe3+ (ferrioxamine B, FOB), was studied, using thermo X-ray diffraction (thermo-XRD) in the temperature range 25-360 degrees C and thermo-FTIR spectroscopy in the temperature range 25 170 degrees C. The effect of pH (4-7.5) on the adsorption was examined. Extensive use of curve-fitting analysis was required due to significant overlapping of the characteristic absorption bands of the various functional groups. Thermo-XRD analysis showed that both DFOB and FOB penetrated into the interlayer space of Ca montmorillonite. FTIR results indicated strong interactions of DFOB within the interlayer, which involved all functional groups (NH3+, secondary amide groups, and hydroxamate groups). In contrast, the folded Fe complex of FOB retained its molecular configuration upon adsorption, and the basal spacing of the clay increased correspondingly. FOB interacted in the interlayer space of the clay, mainly through the NH of the secondary amide groups and NH3+, while the functional groups bound to the central Fe cation remained unchanged. The suspension pH had no significant effect on both DFOB and FOB adsorption at the examined range. Adsorption protected the adsorbates from thermal degradation compared to the nonadsorbed samples up to 105 degrees C. At 170 degrees C both DFOB and FOB were already partially degraded, but to a lesser extent than the nonadsorbed samples. Degradation of the molecules occurred mainly through the hydroxamic groups, which constitute the Fe-chelating center in the hydroxamic siderophore. PMID- 16478267 TI - Identification of a stale-beer-like odorant in extracts of naturally aged beer. AB - For a long time, beer staling has been a prime concern in brewery research. Yet, to improve flavor stability, better knowledge of all chemicals involved is still needed. From our aroma extract dilu-tion analyses (AEDA) applied to naturally aged lager beers emerged an old-beer-like odorant at RICP-SIL 5 CB = 1532 and RIFFAP = 2809, with a FD value close to that of trans-2-nonenal (the well-known cardboard off-flavor found in aged beers). Specific phenol extraction, GC cold trapping, and mass spectrometry (electron impact and chemical ionization) enabled us to identify it as 4-vinylsyringol. Although already mentioned in some fresh beers, this compound had never been highlighted as involved in the aging process of lager beers. PMID- 16478268 TI - Essential oil composition of diploid and tetraploid clones of ginger (Zingiber officinale Roscoe) grown in Australia. AB - Ginger oil, obtained by steam distillation of the rhizome of Zingiber officinale Roscoe, is used in the beverage and fragrance industries. Ginger oil displays considerable compositional diversity, but is typically characterized by a high content of sesquiterpene hydrocarbons, including zingiberene, ar-curcumene, beta bisabolene, and beta-sesquiphellandrene. Australian ginger oil has a reputation for possessing a particular "lemony" aroma, due to its high content of the isomers neral and geranial, often collectively referred to as citral. Fresh rhizomes of 17 clones of Australian ginger, including commercial cultivars and experimental tetraploid clones, were steam distilled 7 weeks post-harvest, and the resulting oils were analyzed by GC-MS. The essential oils of 16 of the 17 clones, including the tetraploid clones and their parent cultivar, were found to be of substantially similar composition. These oils were characterized by very high citral levels (51-71%) and relatively low levels of the sesquiterpene hydrocarbons typical of ginger oil. The citral levels of most of these oils exceeded those previously reported for ginger oils. The neral-to-geranial ratio was shown to be remarkably constant (0.61 +/- 0.01) across all 17 clones. One clone, the cultivar "Jamaican", yielded oil with a substantially different composition, lower citral content and higher levels of sesquiterpene hydrocarbons. Because this cultivar also contains significantly higher concentrations of pungent gingerols, it possesses unique aroma and flavor characteristics, which should be of commercial interest. PMID- 16478269 TI - Chemical composition of volatiles in Sardinian myrtle (Myrtus communis L.) alcoholic extracts and essential oils. AB - The chemical composition of the volatile fraction of myrtle (Myrtus communis L.) alcoholic extracts and essential oils from leaves and berries collected in different places in Sardinia (Italy) was studied. A simple and rapid liquid liquid extraction method was used to isolate volatile compounds from myrtle alcoholic extracts followed by GC and GC-MS analysis allowing the detection of 24 compounds. The volatile fraction was characterized by the terpenes fraction corresponding to that of the essential oils and by a fatty acid ethyl esters fraction. The variation during extraction of the volatile fraction in alcoholic extracts of berries and leaves was evaluated. Essential oils were obtained by hydrodistillation, and the yields were on average 0.52 +/- 0.03% (v/w dried weight) and 0.02 +/- 0.00% for leaves and berries, respectively. The essential oils were analyzed by GC and GC-MS, and a total of 27 components were detected, accounting for 90.6-98.7% of the total essential oil composition. Strong chemical variability depending on the origin of the samples was observed. The major compounds in the essential oils were alpha-pinene (30.0 and 28.5%), 1,8-cineole (28.8 and 15.3%), and limonene (17.5 and 24.1%) in leaves and berries, respectively, and were characterized by the lack of myrtenyl acetate. PMID- 16478270 TI - Application of electron spin resonance spectroscopy and spin probes to investigate the effect of ingredients on changes in wheat dough during heating. AB - The change in microviscosity of the aqueous and lipid phases of wheat flour dough, during heating and subsequent cooling, has been measured using novel spin probes based on the isoindolin-yloxyl structure. The spin probes, water and/or lipid soluble, were used with combinations of dough ingredients: diacetyl tartaric acid ester of monoglycerides (DATEM), salt, yeast, and sodium ascorbate. The lipid soluble probe showed that DATEM does not produce a homogeneous phase with endogenous lipids but is found in a separate, less mobile phase. Also, the lipids were shown not to be involved in the baking process, although DATEM may be incorporated into the gelled starch matrix. The water soluble probe enabled starch gelatinization to be investigated in detail and showed that gelatinization produces a reduction of dielectric constant. The technique is appropriate for the detailed examination of the behavior of different ingredients during baking and also potentially to examine interactions between ingredients and flour components in dough. PMID- 16478271 TI - Stabilizing behavior of soy soluble polysaccharide or high methoxyl pectin in soy protein isolate emulsions at low pH. AB - The stability of emulsions prepared with soy protein isolates was investigated as a function of pH in the presence of two negatively charged polysaccharides: high methoxyl pectin (HMP) and soy soluble polysaccharide (SSPS). Both polysaccharides are composed of a backbone which contains galacturonic acid but, when added to soy protein isolate-stabilized emulsions, SSPS showed a different behavior than that of HMP. At neutral pH and above a critical concentration of stabilizer (0.05%), HMP caused flocculation of the emulsion droplets via a depletion mechanism. On the other hand, the emulsions containing a similar amount of SSPS did not show creaming or flocculation. At acidic pH (<4.0) the addition of pectin caused extensive droplet aggregation, while no aggregation was observed with the addition of SSPS. The differences in the stabilization behavior between the two polysaccharides can be attributed to their differences in charge, neutral sugars side chains, and molecular weight. PMID- 16478272 TI - Biosynthesis of trans-2-hexenal in response to wounding in strawberry fruit. AB - Wounded strawberry fruit produces a diverse group of volatile compounds including aldehydes, alcohols, and esters derived from the lipoxygenase (LOX) and hydroperoxide lyase (HPL) pathways. Because the wound volatiles may play an important role in plant-fungal interaction, the goal of this study was to develop a greater understanding about the biosynthesis of the major wound volatile, trans 2-hexenal (t-2-H), produced by strawberry fruit upon wounding. To that end, composition and quantity of total and free fatty acids of control and wounded strawberry fruit were analyzed. In addition, activities of the key enzymes, LOX and HPL, and production of C6 aldehydes were determined. Intact strawberry fruit did not produce detectable t-2-H which is derived from alpha-linolenic acid (18:3). However, in response to wounding by bruising, strawberry fruit emitted t 2-H and its precursor cis-3-hexenal (c-3-H). The level of total lipid 18:3 in the fruit increased 2-fold in response to wounding, whereas free 18:3 declined slightly ( approximately 30%). At 10 min following wounding, fruit exhibited a 25% increase in LOX activity, which leads to the production of 13 hydroperoxyoctadecatrienoic acid (13-HPOT) from 18:3. The activity of HPL, which catalyzes formation of cis-3-hexenal from 13-HPOT, increased 2-fold by 10 min after wounding. Thus, during a 15 min period after wounding, free 18:3 substrate availability and the activity of two key enzymes, LOX and HPL, changed in a manner consistent with increased c-3-H and t-2-H biosynthesis. PMID- 16478273 TI - Antigenic stability of pecan [Carya illinoinensis (Wangenh.) K. Koch] proteins: effects of thermal treatments and in vitro digestion. AB - Rabbit polyclonal antibody-based inhibition ELISA as well as immunoblotting analyses of proteins extracted from variously processed pecans (cv. Desirable) indicate that pecan proteins are antigenically stable. Pecan antigens were more sensitive to moist heat than dry heat processing treatments. SDS-PAGE and immunoblotting analysis of the native and heat-denatured proteins that were previously subjected to in vitro simulated gastric fluid digestions indicate that stable antigenic peptides were produced. Both enzyme-to-substrate ratio and digestion time were influential in determining the stability of pecan polypeptides. The stable antigenic polypeptides may serve as useful markers in developing assays suitable for the detection of trace amounts of pecans in foods. PMID- 16478274 TI - Speciation of phytate ion in aqueous solution. Sequestering ability toward mercury(II) cation in NaClaq at different ionic strengths. AB - As a contribution to understanding the speciation of mercury in the environment and to the study of the sequestering ability of phytate (Phy) toward heavy metal and organometal cations, this paper describes the results of an investigation (at t = 25 degrees C by potentiometry, ISE-H+ glass electrode) of its interactions with mercury(II) cation in NaCl aqueous solutions at different ionic strengths (I = 0.15 and 1.0 mol L(-1)), in the pH range 2.5 < or = pH < or = 9.5 and considering metal-to-ligand ratios of 1:1 < or = Hg/Phy < or = 4:1. The formation of 11 HgiHjPhy(12-2i-j)(- species with i = 1 and 0 < or = j < or = 7 and i = 2 and 0 < or = j < or = 2 was observed. Their complex formation constant values proved to be fairly dependent on ionic strength. The speciation of phytic acid and mercury(II) is also dependent on the metal-to-ligand ratio; the dependence of the stability of phytate-mercury(II) species on the phytate protonation step was modeled, and an empirical predictive relationship was proposed. From the results obtained, phytate has very good sequestering ability toward Hg2+, even in the presence of considerable excesses of chloride ion, that is, another ligand strongly interacting with mercury; this supports future studies both on the use of plants that naturally synthesize it for phytoremediation purposes and on its direct application in remediation techniques. PMID- 16478275 TI - Urinary excretion of black raspberry (Rubus occidentalis) anthocyanins and their metabolites. AB - Anthocyanins are the most abundant phenolic compounds, widely distributed in fruits and vegetables, and exhibit potent antioxidant capacity. Humans ingest a significant amount of anthocyanins in the daily diet. The objective of the current study was to examine human absorption and metabolism of black raspberry anthocyanins when administered at high doses (2.69 +/- 0.085 g/day). Ten healthy men consumed 45 g of freeze-dried black raspberries daily for 1 week. Urine samples were collected over a 12 h period in 4 h intervals at day 1 and day 7. Urinary anthocyanins were analyzed by high-performance liquid chromatography coupled to a photodiode array detector and a tandem mass spectrometer using precursor ion and product ion analyses. Anthocyanins were excreted in intact forms and metabolized into methylated derivatives in human urine. The urinary excretion of anthocyanins reached a maximum concentration (1091.8 +/- 1081.3 pmol/L, n = 10) during the 4-8 h period after black raspberry ingestion. As compared to the anthocyanin distribution in black raspberries, urinary cyanidin 3 xylosylrutinoside was detected at a higher concentration than that of cyanidin-3 rutinoside. PMID- 16478276 TI - Biosynthesis of mono- and sesquiterpenes in strawberry fruits and foliage: 2H labeling studies. AB - The biosynthesis of the monoterpene (S)-linalool and the sesquiterpene trans-(S) nerolidol in fruits of Fragaria x ananassa Duch. cv. Eros and Florence and of the monoterpene (-)-alpha-pinene in Fragaria vesca was investigated by in vivo feeding experiments with [5,5-2H2]mevalonic acid lactone (d2-MVL) and [5,5-2H2]-1 deoxy-d-xylulose (d2-DOX). The feeding experiments indicate that (S)-linalool and trans-(S)-nerolidol in Fragaria x ananassa Duch. and (-)-alpha-pinene in F. vesca are exclusively synthesized via the cytosolic mevalonic acid pathway without any contribution from the plastidial 1-deoxy-D-xylulose/2-C-methyl-D-erythritol 4 phosphate (DOXP/MEP) route. Inhibition experiments revealed that even the presence of mevastatin, an export of plastid-derived isopentyl diphosphate/dimethylallyl diphosphate, cannot be induced. However, the enantioselective analysis shows that in Fragaria x ananassa Duch. cv. Eros and Florence both linalool enantiomers are present and that only (S)-linalool is labeled after administration of d2-MVL. Therefore, the origin of (R)-linalool in these fruits remains unknown. Contrarily, in Fragaria x ananassa Duch. foliage (R)-linalool is the dominant enantiomer. Feeding experiments revealed an incorporation of d2-MVL and d2-DOX at equal rates exclusively into (S)-linalool. Only in F. vesca foliage, where (R)-linalool is present at high enantiomeric purity (ee > 90%), is a de novo biosynthesis of the (R)-enantiomer via the DOXP/MEP pathway detectable. These results demonstrate a complex intraplant variation of (R)- and (S)-linalool biosynthesis via the cytosolic and plastidial route. PMID- 16478277 TI - Two-phase flavonoid formation in developing strawberry (Fragaria x ananassa) fruit. AB - Flavonoids are important secondary metabolites in strawberry as they fulfill a wide variety of physiological functions. In addition, they are beneficial for human health. Previous studies have shown for selected enzymes from the flavonoid pathway that flavonoid biosynthesis shows two peaks during fruit development. We provide optimized protocols for the determination of the activities of the key flavonoid enzymes: phenylalanine ammonia lyase, chalcone synthase/chalcone isomerase, flavanone 3-hydroxylase, dihydroflavonol 4-reductase, flavonol synthase, flavonoid 3-O-glucosyltransferase, and flavonoid 7-O glucosyltransferase. Using these protocols we were able to demonstrate two distinct activity peaks during fruit ripening at early and late developmental stages for all enzymes with the exception of flavonol synthase. The first activity peak corresponds to the formation of flavanols, while the second peak is clearly related to anthocyanin and flavonol accumulation. The results indicate that flavonoid 3-O-glucosyltransferase activity is not essential for redirection from flavanol to anthocyanin formation in strawberry. PMID- 16478278 TI - Kinetic model for studying the effect of quercetin on cholesterol oxidation during heating. AB - Inhibition of the heat-induced cholesterol oxidation at 150 degrees C by incorporation of quercetin was kinetically studied. Results showed that without quercetin, the cholesterol oxidation products (COPs) concentration increased with increasing heating time. A low amount (0.002%, w/w) of quercetin was effective in inhibiting the formation of COPs during the initial heating period (< or =30 min) at 150 degrees C. However, after prolonged heating (30-120 min), a low antioxidant activity was observed because of the degradation of quercetin. When using nonlinear regression models for kinetic study of cholesterol oxidation in the absence of quercetin, the epoxidation showed the highest rate constant (h(-1) = 683.1), followed by free radical chain reaction (h(-1) = 453.5), reduction (h( 1) = 290.3), dehydration (h(-1) = 155.5), triol dehydrogenation (h(-1) = 5.35), dehydrogenation (h(-1) = 0.68), thermal degradation (h(-1) = 0.66), and triol formation (h(-1) = 0.38). However, in the presence of quercetin, the reaction rate constants (h(-1)) for epoxidation (551.4), free radical chain reaction (111.7), and thermal degradation (0.28) were reduced greatly. The kinetic model developed in this study can be used to predict the inhibition of COPs by quercetin during the heating of cholesterol. PMID- 16478279 TI - Amylose-lipid complexes as controlled lipid release agents during starch gelatinization and pasting. AB - The effect of amylose-lipid (AM-L) complexes consisting of amylose populations with different peak degrees of polymerization (DP) and complexed with glyceryl monostearate (GMS) or docosanoic acid (C22) on the pasting properties of wheat and rice starches was evaluated with a rapid visco analyzer (RVA). AM-L complexes were formed by both (i) addition of lipids to amylose fractions with peak DP 20, 60, 400, or 950 at 60 degrees C or (ii) potato phosphorylase-catalyzed amylose synthesis in the presence of lipids. All AM-L complexes affected pasting properties in line with their dissociation characteristics. AM-L complexes therefore have potential as "controlled lipid release agents" with effects markedly different from those observable with emulsifier addition in starch pasting. More in particular, short chain AM-L complexes resulted in a starch pasting behavior comparable to that of cross-linked starch, as evidenced by reduced granule swelling, good viscosity stability in conditions of high temperature and shear, and a stable cold paste viscosity. PMID- 16478280 TI - Enterocyte and M-cell transport of native and heat-denatured bovine beta lactoglobulin: significance of heat denaturation. AB - The three-dimensional structure, digestibility, and immunological properties of bovine beta-lactoglobulin (beta-lg) are modified by heat treatments used in processing of liquid milk products. Because it is not known if such treatments also modify the intestinal transport properties of beta-lg, the transport of native and heat-denatured bovine beta-lg was investigated in experimental cell models using Caco-2 cells and M cells. Transport of beta-lg labeled with a fluorescent marker was followed with fluorometric measurements, electrophoretic analyses, and fluorescence microscopy. The data show that both cell types transported native beta-lg more efficiently than they did heat-denatured beta-lg. In addition, M cells transported native beta-lg more than Caco-2 cells. Transport of native and heat-denatured beta-lg was transcellular. The electrophoretic data also suggest that heat-denatured beta-lg may have degraded more than native beta lg during the transport. PMID- 16478281 TI - Heat markers and quality indexes of industrially heat-treated [15N] milk protein measured in rats. AB - To determine the bioavailability of industrially heat-treated milk proteins, male Wistar rats were given [15N]-labeled meals containing either nonheated-micellar casein (CAS), milk soluble protein isolate (MSPI), and microfiltered milk (MF)-or heated products-"high temperature short time" pasteurized (HTST), "higher temperature, shorter time" pasteurized (HHST), ultrahigh temperature-treated (UHT), and spray-dried (SPRAY) milks. The postprandial distribution of dietary nitrogen was measured in the splanchnic area and urea. Digestibility was around 96% except for SPRAY (94%) and MSPI (98%). Ingested nitrogen recovered in the splanchnic bed was 19.3% for SPRAY, 16.7% for MF, and around 14-15% for other products. Deamination of dietary nitrogen reached 21.2, 20.6, and 18.2% of ingested nitrogen for MSPI, SPRAY, and CAS, respectively, and around 14-16% for other products. In our model, only spray drying led to a significant increase of splanchnic extraction. Moreover, the biological value of purified protein fractions appeared to be lower than that seen in products containing total milk protein. PMID- 16478282 TI - Iron bioavailability of hemoglobin from soy root nodules using a Caco-2 cell culture model. AB - Heme iron has been identified in many plant sources-most commonly in the root nodules of leguminous plants, such as soy. Our objective was to test the effectiveness of soy root nodule (SRN) and purified soy hemoglobin (LHb) in improving iron bioavailability using an in vitro Caco-2 cell model, with ferritin response as the bioavailability index. We assessed bioavailability of iron from LHb (either partially purified (LHbA) or purified (LHbD)) with and without food matrix and compared it with that from bovine hemoglobin (BHb), ferrous sulfate (FeSO4), or SRN. Bioavailability of each treatment was normalized to 100% of the FeSO4 treatment. When iron sources were tested alone (100 ug iron/mL), ferritin synthesis by LHbD and BHb were 19% (P > 0.05) and 113% (P < 0.001) higher than FeSO4, respectively. However, when iron sources were used for fortification of maize tortillas (50 ppm), LHbA and BHb showed similar bioavailability, being 27% (P < 0.05) and 33% (P < 0.05) higher than FeSO4. Heat treatment had no effect on heme iron but had a significant reduction on FeSO4 bioavailability. Adding heme (LHbA) iron with nonheme (FeSO4) had no enhancement on nonheme iron absorption. Our data suggest that heme iron from plant sources may be a novel value-added product that can provide highly bioavailable iron as a food fortificant. PMID- 16478283 TI - Verification of compliance with organic meat production standards by detection of permitted and nonpermitted uses of veterinary medicines (tetracycline antibiotics). AB - In the production of "organic" meat, one of the controlled processes is the use of veterinary drugs. Strict standards are in place as to when and how such drugs may be used. Therefore, the aim of this project was to determine whether it was possible to distinguish between a single therapeutic dose of a tetracycline (permitted under the standards) and both multiple therapeutic dosing and prophylactic dosing (not permitted). This comprised an evaluation of (i) pigs that were treated with oxytetracycline and (ii) chickens dosed with two different tetracycline antibiotics (oxytetracycline and chlortetracycline). The methodology described, using bone sectioning and examination under ultraviolet illumination (either direct observation or fluorescent microscopy), allows samples from animals that have been treated with different dosing regimes (a single therapeutic dose, two successive therapeutic doses, and long-term, low-level "prophylactic" dosing) to be assessed for compliance with organic farming regulations. Validation of the methodology by blind checks of unknown samples by a second operator has been successfully performed, and validation results are presented. The developed methodology has been shown to be applicable to a variety of species and a selection of tetracycline drugs. PMID- 16478284 TI - Production of 7-epi-pectenotoxin-2 seco acid and assessment of its acute toxicity to mice. AB - Pectenotoxins (PTXs) accumulate in shellfish feeding on dinoflagellates of the genus Dinophysis, so that humans can be exposed to these toxins through shellfish consumption. Some PTXs are toxic to experimental animals, whereas others are of much lower toxicity. Pectenotoxin-2, the most abundant PTX from most Dinophysis spp., is rapidly metabolized by most shellfish to a mixture of pectenotoxin-2 seco acid (2) and 7-epi-pectenotoxin-2 seco acid (1). A mixture of 1 and 2 was produced during purification of an extract from in vitro enzymatic hydrolysis of pectenotoxin-2. These were separated by preparative HPLC, and the structure of 1 was confirmed by one- and two-dimensional 1H and 13C NMR spectroscopy and LC-MS3 analyses. No toxic changes were recorded in mice injected intraperitoneally with 1 or 2 at a dose of 5000 microg/kg. PTX seco acids are therefore unlikely to be of consequence to human consumers at the concentrations found in contaminated shellfish. PMID- 16478286 TI - Media portrayal of mental illness and its treatments: what effect does it have on people with mental illness? AB - This article reviews dominant media portrayals of mental illness, the mentally ill and mental health interventions, and examines what social, emotional and treatment-related effects these may have. Studies consistently show that both entertainment and news media provide overwhelmingly dramatic and distorted images of mental illness that emphasise dangerousness, criminality and unpredictability. They also model negative reactions to the mentally ill, including fear, rejection, derision and ridicule. The consequences of negative media images for people who have a mental illness are profound. They impair self-esteem, help seeking behaviours, medication adherence and overall recovery. Mental health advocates blame the media for promoting stigma and discrimination toward people with a mental illness. However, the media may also be an important ally in challenging public prejudices, initiating public debate, and projecting positive, human interest stories about people who live with mental illness. Media lobbying and press liaison should take on a central role for mental health professionals, not only as a way of speaking out for patients who may not be able to speak out for themselves, but as a means of improving public education and awareness. Also, given the consistency of research findings in this field, it may now be time to shift attention away from further cataloguing of media representations of mental illness to the more challenging prospect of how to use the media to improve the life chances and recovery possibilities for the one in four people living with mental disorders. PMID- 16478285 TI - Is there a future for cyclo-oxygenase inhibitors in Alzheimer's disease? AB - Several epidemiological studies have indicated that the long-term use of NSAIDs, most of which are cyclo-oxygenase (COX) inhibitors, may reduce the risk of Alzheimer's disease. For this reason, anti-inflammatory COX-inhibiting NSAIDs have received increased attention in experimental and therapeutic trials for Alzheimer's disease. However, several recent efforts attempting to demonstrate a therapeutic effect of NSAIDs in Alzheimer's disease have largely failed. Clinicians and scientists currently believe that this lack of success may be attributable to two key problems: (i) clinical trials of NSAIDs have been conducted in patients with late-stage Alzheimer's disease, wherein advanced neurodegeneration may be refractory to anti-inflammatory drug treatment; and (ii) it is not known which of the large family of NSAIDs (i.e. COX-1, COX-2 or mixed inhibitors) is most efficacious in preventing Alzheimer's disease. The wide list of putative functions for COX in the brain, and the significant functional heterogeneity of NSAIDs, which appear to influence the beta-amyloid (Abeta) neuropathology associated with Alzheimer's disease via both COX-dependent and COX independent pathways, complicate the interpretation of the mechanisms through which COX-inhibiting NSAIDs may beneficially influence Alzheimer's disease. As discussed in this review, for patients at high risk of developing Alzheimer's disease (e.g. those with mild cognitive impairment), preventative treatment with COX-inhibiting NSAIDs may ultimately represent a viable strategy in the management of clinical Alzheimer's disease. However, the recent evidence showing an increased risk of major cardiovascular events among patients treated with certain COX-1 and COX-2 inhibitors leaves many questions unanswered. We suggest that further investigation into the physiological role(s) of COXs in normal health and in disease conditions, and the identification of safer and better tolerated COX inhibitors, will provide renewed impetus to the application of anti inflammatory strategies for the prevention and treatment of Alzheimer's disease. PMID- 16478289 TI - SSRIs and upper gastrointestinal bleeding: what is known and how should it influence prescribing? AB - SSRIs have achieved a high usage rate in the treatment of depression because of a similar efficacy to TCAs and a favourable safety and tolerability profile. However, SSRI use has been associated with bleeding. We reviewed the epidemiological evidence on the association between SSRI use alone and the risk of upper gastrointestinal bleeding, and on synergistic effects reported with other commonly used drugs that can also cause bleeding.A literature search identified four studies of SSRI use and risk for upper gastrointestinal bleeding and a further two studies of SSRI use and bleeding in general, including upper gastrointestinal bleeding. The available evidence indicates quite convincingly that SSRI use may play a causal role in upper gastrointestinal bleeding and that these drugs may act synergistically with other bleeding risk-increasing medications such as NSAIDs or low-dose aspirin. Assuming a causal role of SSRIs, reported excess gastrointestinal bleedings attributable to SSRI use was reported to be 3.1 per 1000 treatment years, 4.1 per 1000 treatment years among octogenarians and 11.7 per 1000 treatment years among persons with prior upper gastrointestinal bleeding. These non-negligible risks warrant that prescribing doctors consider strategies on the individual level to reduce the likelihood of an upper gastrointestinal adverse event. Patients at particular risk include those with previous ulcers or gastrointestinal bleeding, the elderly and those with certain concurrent illnesses and/or high-risk comedications. Suggested strategies include alternatives to SSRI use, prescribing of less gastrotoxic NSAIDs or co-prescribing of gastroprotective drugs. Patients should be informed about the likelihood of possible upper gastrointestinal bleeding and high-risk patients should be followed closely. PMID- 16478288 TI - Hormone-related headache: pathophysiology and treatment. AB - Epidemiological data suggest a link between migraine and the female sex hormones. Indeed, it is known that estrogen affects various brain functions, including pain perception. The prevalence of migraine is similar in boys and girls before puberty, but is 3-fold higher in postpubertal females compared with males. Migraine attacks in women are more likely to occur in the perimenstrual period and occur exclusively so in some women. The acute treatment of menstrual migraine is similar to that of non-menstrually related attacks, but the response to treatment may be less favourable. Perimenstrual prophylaxis, with NSAIDs, triptans or estradiol, is effective in decreasing attack frequency and severity. The use of oral contraceptives (OCs) may change migraine frequency and severity. Since both migraine and hormonal contraceptive use are risk factors for ischaemic stroke, the use of OCs in women who experience migraine should be made only after consideration of the benefit-risk ratio. Migraine typically, but not invariably, improves during the last two trimesters of pregnancy, and may worsen in the postpartum period. When using drugs to treat migraine during pregnancy, potential risks to the mother and fetus should be considered. The prevalence of migraine decreases with advancing age and it improves in many, but not all, women after the menopause. However, in the perimenopausal period, migraine may worsen as a result of fluctuations in estrogen levels. Reducing the estrogen dose and changing the estrogen type or the route of administration of hormone replacement therapy (HRT) from oral to transdermal may reduce headache. Migraine is not a risk factor for stroke in postmenopausal women. When considering symptomatic HRT for postmenopausal migraneurs, the usual indications and contraindications should be applied. HRT may also exacerbate migraine. PMID- 16478287 TI - Attention-deficit hyperactivity disorder in girls: epidemiology and management. AB - Attention-deficit hyperactivity disorder (ADHD) in girls is a topic of growing research and clinical interest. For many years, girls with ADHD have been ignored and overshadowed by hyperkinetic and impulsive boys, but they are now attracting interest in an effort to understand the similarities and differences in the prevalence, symptoms, familial risk, comorbidities and treatment of ADHD in the two sexes. A review of past and current literature finds that the symptoms of ADHD are not sex specific, but that identification of girls with ADHD is hampered by parental and teacher bias, and confusion. Girls are more likely to be inattentive without being hyperactive or impulsive, compared with boys. Girls and boys share the same familial risk patterns, as well as similar, although not identical, comorbidity or impairment patterns. The risk of non-treatment is as great in girls as it is in boys; up to 70-80% of identified children will have persistent symptoms and impairment that extends into adolescence and adulthood. Treatment modalities are equally effective in girls and boys. Stimulants, non stimulants and behavioural modalities are the mainstays of effective treatment. PMID- 16478291 TI - Spotlight on escitalopram in the management of major depressive disorder. AB - Escitalopram (Cipralex), Lexapro), the active S-enantiomer of the racemic SSRI citalopram (RS-citalopram), is a highly selective inhibitor of the serotonin transporter protein. It possesses a rapid onset of antidepressant activity, and is an effective and generally well tolerated treatment for moderate-to-severe major depressive disorder (MDD). Pooled analyses from an extensive clinical trial database suggest that escitalopram is consistently more effective than citalopram in moderate-to-severe MDD. Preliminary studies suggest that escitalopram is as effective as other SSRIs and the extended-release (XR) formulation of the serotonin/noradrenaline (norepinephrine) reuptake inhibitor venlafaxine, and may have cost-effectiveness and cost-utility advantages. However, additional longer term, comparative studies evaluating specific efficacy, tolerability, health related quality of life and economic indices would be helpful in definitively positioning escitalopram relative to these other agents in the treatment of MDD. Nevertheless, available clinical and pharmacoeconomic data indicate that escitalopram is an effective first-line option in the management of patients with MDD. PMID- 16478292 TI - Augmenting strategies in clozapine-resistant schizophrenia. PMID- 16478290 TI - Biological markers of age-related memory deficits: treatment of senescent physiology. AB - In humans, age-related memory impairments begin in mid-life and cognitive weakening continues with advancing age. An important aspect of defining memory decline is the distinction between dementia as a result of neurological diseases, such as Alzheimer's disease, and memory loss not specifically associated with disease. Within the population of elderly without dementia, there is considerable variability in memory. This variability is likely to be a result of the interaction of genetic make-up and environment, which influences several processes for cell maintenance and repair including oxidative damage and cholesterol metabolism, leading to disruption of Ca(2+) homeostasis, and ultimately Ca(2+)-dependent processes that underlie memory. In humans, several methods have been employed to distinguish biological markers of aging that may predict cognitive decline. Memory deficits associated with normal aging and Alzheimer's disease have been linked to a decrease in the volume of brain structures, such as the hippocampus and to genetic markers, such as apolipoprotein E. In this regard, examination of CSF for biomarkers of disease can help in differentiating normal aging from Alzheimer's disease. Measures of oxidative stress and cholesterol in plasma correlate with memory deficits; research suggests that treatments that reduce oxidative stress or cholesterol through exercise, diet or the use of antioxidant vitamins may delay cognitive decline.Nevertheless, to date, very little treatment is available to reverse memory deficits in later life. In this regard it is important to identify individuals at risk for memory deficits in order to discriminate different mechanisms of brain aging and develop treatments. Considerable effort is driving research to develop accurate biological markers of brain aging. In turn, these markers will provide information on mechanisms of aging and cognitive decline and point to potential treatments. Accordingly, the effectiveness of treatment needs to be verified for both cognitive changes and biological markers that are specific for age-related memory deficits. PMID- 16478293 TI - Nicotinamide protects against ethanol-induced apoptotic neurodegeneration in the developing mouse brain. AB - BACKGROUND: Exposure to alcohol during brain development may cause a neurological syndrome called fetal alcohol syndrome (FAS). Ethanol induces apoptotic neuronal death at specific developmental stages, particularly during the brain-growth spurt, which occurs from the beginning of third trimester of gestation and continues for several years after birth in humans, whilst occurring in the first two postnatal weeks in mice. Administration of a single dose of ethanol in 7-d postnatal (P7) mice triggers activation of caspase-3 and widespread apoptotic neuronal death in the forebrain, providing a possible explanation for the microencephaly observed in human FAS. The present study was aimed at determining whether nicotinamide may prevent ethanol-induced neurodegeneration. METHODS AND FINDINGS: P7 mice were treated with a single dose of ethanol (5 g/kg), and nicotinamide was administered from 0 h to 8 h after ethanol exposure. The effects of nicotinamide on ethanol-induced activation of caspase-3 and release of cytochrome-c from the mitochondria were analyzed by Western blot (n = 4-7/group). Density of Fluoro-Jade B-positive cells and NeuN-positive cells was determined in the cingulated cortex, CA1 region of the hippocampus, and lateral dorsal nucleus of the thalamus (n = 5-6/group). Open field, plus maze, and fear conditioning tests were used to study the behavior in adult mice (n = 31-34/group). Nicotinamide reduced the activation of caspase-3 (85.14 +/- 4.1%) and the release of cytochrome-c (80.78 +/- 4.39%) in postnatal mouse forebrain, too. Nicotinamide prevented also the ethanol-induced increase of apoptosis. We demonstrated that ethanol-exposed mice showed impaired performance in the fear conditioning test and increased activity in the open field and in the plus maze. Administration of nicotinamide prevented all these behavioral abnormalities in ethanol-exposed mice. CONCLUSIONS: Our findings indicate that nicotinamide can prevent some of the deleterious effects of ethanol on the developing mouse brain when given shortly after ethanol exposure. These results suggest that nicotinamide, which has been used in humans for the treatment of diabetes and bullous pemphigoid, may hold promise as a preventive therapy of FAS. PMID- 16478294 TI - Evidence-based prescribing of statins: a developing world perspective. PMID- 16478297 TI - Ultraviolet radiation attenuates thrombospondin 1 expression via PI3K-Akt activation in human keratinocytes. AB - Thrombospondin 1 (TSP1) is an extracellular glycoprotein and a recognized inhibitor of angiogenesis. Recent studies have demonstrated that UV radiation induces an angiogenic switch, by which it alters the balance between pro- and anti-angiogenic factors in the skin. Here we describe the effects of acute UV exposure on TSP1 expression in human skin epidermis, primary keratinocytes and the epidermal cell line HaCaT. We found that protein and mRNA expressions of TSP1 are significantly reduced in human skin in vivo and in keratinocytes in vitro by a single UV exposure. In human skin and keratinocytes, UV exposure induced the phosphorylation of Akt, a downstream target of the PI3K pathways. Specific inhibitors of PI3K, wortmannin and LY294002, completely blocked Akt activation and UV-induced TSP1 downregulation in keratinocytes. We showed that a specific Akt phosphorylation inhibitor and small interfering RNA-mediated Akt depletion were also blocked by UV-induced TSP1 downregulation in keratinocytes. In conclusion, our findings demonstrate that acute UV exposure downregulates TSP1 expression via PI3K-Akt activation in human keratinocytes. These novel findings may help us understand the regulatory mechanisms of UV-induced skin angiogenesis. PMID- 16478295 TI - Enrolling women into HIV preventive vaccine trials: an ethical imperative but a logistical challenge. PMID- 16478298 TI - Rate measurement of the reaction of CF2Cl radicals with O2. AB - We have studied the association reaction of the CF(2)Cl radicals with O(2) in presence of N(2). The infrared multiple photon dissociation (IRMPD) technique with a homemade TEA CO(2) laser was used for the CF(2)Cl radical generation and the vibrational chemiluminiscence technique was set up for the study of the reaction kinetics. The time-resolved IR fluorescence of the vibrationally excited CF(2)O photoproduct was used to measure the disappearance rate of these radicals. A kinetic mechanism is presented to account for the rate of production of CF(2)O(*). The CF(2)Cl radical association reaction rate with O(2), evidence of a direct channel of photoproduct formation and its reaction rate, and the CF(2)O(*) collisional deactivation rate have been obtained. PMID- 16478299 TI - Tracing the identity of objects. AB - This article considers how people judge the identity of objects (e.g., how people decide that a description of an object at one time, t(0), belongs to the same object as a description of it at another time, t(1)). The authors propose a causal continuer model for these judgments, based on an earlier theory by Nozick (1981). According to this model, the 2 descriptions belong to the same object if (a) the object at t(1) is among those that are causally close enough to be genuine continuers of the original and (b) it is the closest of these close enough contenders. A quantitative version of the model makes accurate predictions about judgments of which a pair of objects is identical to an original (Experiments 1 and 2). The model makes correct qualitative predictions about identity across radical disassembly (Experiment 1) as well as more ordinary transformations (Experiments 2 and 3). PMID- 16478300 TI - Memory hazard functions: a vehicle for theory development and test. AB - A framework is developed to rigorously test an entire class of memory retention functions by examining hazard properties. Evidence is provided that the memory hazard function is not monotonically decreasing. Yet most of the proposals for retention functions, which have emerged from the psychological literature, imply that memory hazard is monotonically decreasing over the entire temporal range. Furthermore, the few remaining proposals, that do not have monotonically decreasing hazard, have difficulty fitting data over both short-term and long term intervals. A new 2-trace hazard model is developed that successfully circumvents these difficulties. This new model is used to account for the size of memory span and the time course of proactive and retroactive interference effects. The model can fit the retention characteristics of H. M., the famous amnesic patient, as well as normal experimental participants. The model is also used to account for the time course of the misinformation effect. PMID- 16478301 TI - Global model analysis by parameter space partitioning. AB - To model behavior, scientists need to know how models behave. This means learning what other behaviors a model can produce besides the one generated by participants in an experiment. This is a difficult problem because of the complexity of psychological models (e.g., their many parameters) and because the behavioral precision of models (e.g., interval-scale performance) often mismatches their testable precision in experiments, where qualitative, ordinal predictions are the norm. Parameter space partitioning is a solution that evaluates model performance at a qualitative level. There exists a partition on the model's parameter space that divides it into regions that correspond to each data pattern. Three application examples demonstrate its potential and versatility for studying the global behavior of psychological models. PMID- 16478302 TI - Groups as epistemic providers: need for closure and the unfolding of group centrism. AB - Theory and research are presented relating the need for cognitive closure to major facets of group behavior. It is suggested that a high need for closure, whether it is based on members' disposition or the situation, contributes to the emergence of a behavioral syndrome describable as group-centrism--a pattern that includes pressures to opinion uniformity, encouragement of autocratic leadership, in-group favoritism, rejection of deviates, resistance to change, conservatism, and the perpetuation of group norms. These theoretical predictions are borne out by laboratory and field research in diverse settings. PMID- 16478303 TI - Toward a comprehensive model of antisocial development: a dynamic systems approach. AB - The purpose of this article is to develop a preliminary comprehensive model of antisocial development based on dynamic systems principles. The model is built on the foundations of behavioral research on coercion theory. First, the authors focus on the principles of multistability, feedback, and nonlinear causality to reconceptualize real-time parent-child and peer processes. Second, they model the mechanisms by which these real-time processes give rise to negative developmental outcomes, which in turn feed back to determine real-time interactions. Third, they examine mechanisms of change and stability in early- and late-onset antisocial trajectories. Finally, novel clinical designs and predictions are introduced. The authors highlight new predictions and present studies that have tested aspects of the model PMID- 16478304 TI - The causal asymmetry. AB - It is hypothesized that there is a pervasive and fundamental bias in humans' understanding of physical causation: Once the roles of cause and effect are assigned to objects in interactions, people tend to overestimate the strength and importance of the causal object and underestimate that of the effect object in bringing about the outcome. This bias is termed the causal asymmetry. Evidence for this bias is reviewed in several domains, including visual impressions of causal relations, reasoning about Newton's third law in naive physics problems, concepts underlying linguistic expressions of causality, and research in causal judgment from contingency information. Although there might be an equivalent to the causal asymmetry in the domain of social causality, there are too many uncertainties in the evidence for conclusions to be drawn. PMID- 16478305 TI - The race model inequality: interpreting a geometric measure of the amount of violation. AB - An inequality by J. O. Miller (1982) has become the standard tool to test the race model for redundant signals reaction times (RTs), as an alternative to a neural summation mechanism. It stipulates that the RT distribution function to redundant stimuli is never larger than the sum of the distribution functions for 2 single stimuli. When many different experimental conditions are to be compared, a numerical index of violation is very desirable. Widespread practice is to take a certain area with contours defined by the distribution functions for single and redundant stimuli. Here this area is shown to equal the difference between 2 mean RT values. This result provides an intuitive interpretation of the index and makes it amenable to simple statistical testing. An extension of this approach to 3 redundant signals is presented. PMID- 16478306 TI - Tests of multiplicative models in psychology: a case study using the unified theory of implicit attitudes, stereotypes, self-esteem, and self-concept. AB - Theories that posit multiplicative relationships between variables are common in psychology. A. G. Greenwald et al. recently presented a theory that explicated relationships between group identification, group attitudes, and self-esteem. Their theory posits a multiplicative relationship between concepts when predicting a criterion variable. Greenwald et al. suggested analytic strategies to test their multiplicative model that researchers might assume are appropriate for testing multiplicative models more generally. The theory and analytic strategies of Greenwald et al. are used as a case study to show the strong measurement assumptions that underlie certain tests of multiplicative models. It is shown that the approach used by Greenwald et al. can lead to declarations of theoretical support when the theory is wrong as well as rejection of the theory when the theory is correct. A simple strategy for testing multiplicative models that makes weaker measurement assumptions than the strategy proposed by Greenwald et al. is suggested and discussed. PMID- 16478310 TI - Parallel distributed processing and lexical-semantic effects in visual word recognition: are a few stages necessary? AB - D. C. Plaut and J. R. Booth (2000) presented a parallel distributed processing model that purports to simulate human lexical decision performance. This model (and D. C. Plaut, 1995) offers a single mechanism account of the pattern of factor effects on reaction time (RT) between semantic priming, word frequency, and stimulus quality without requiring a stages-of-processing account of additive effects. Three problems are discussed. First, no evidence is provided that this model can discriminate between words and nonwords with the same orthographic structure and still produce the pattern of factor effects on RT it currently claims to produce. Second, the level of representation used by the model to make a lexical decision is inconsistent with what is known about how skilled readers with damage to their semantic system make word/nonword discriminations. Finally, there are a number of results that are difficult to reconcile with the single mechanism account. The authors' preference is to retain the stages-of-processing account. PMID- 16478313 TI - Seeing, acting, understanding: motor resonance in language comprehension. AB - Observing actions and understanding sentences about actions activates corresponding motor processes in the observer-comprehender. In 5 experiments, the authors addressed 2 novel questions regarding language-based motor resonance. The 1st question asks whether visual motion that is associated with an action produces motor resonance in sentence comprehension. The 2nd question asks whether motor resonance is modulated during sentence comprehension. The authors' experiments provide an affirmative response to both questions. A rotating visual stimulus affects both actual manual rotation and the comprehension of manual rotation sentences. Motor resonance is modulated by the linguistic input and is a rather immediate and localized phenomenon. The results are discussed in the context of theories of action observation and mental simulation. PMID- 16478314 TI - Tracking the mind during reading: the influence of past, present, and future words on fixation durations. AB - Reading requires the orchestration of visual, attentional, language-related, and oculomotor processing constraints. This study replicates previous effects of frequency, predictability, and length of fixated words on fixation durations in natural reading and demonstrates new effects of these variables related to 144 sentences. Such evidence for distributed processing of words across fixation durations challenges psycholinguistic immediacy-of-processing and eye-mind assumptions. Most of the time the mind processes several words in parallel at different perceptual and cognitive levels. Eye movements can help to unravel these processes. PMID- 16478315 TI - The intricate relationships between monitoring and control in metacognition: lessons for the cause-and-effect relation between subjective experience and behavior. AB - Do we run away because we are frightened, or are we frightened because we run away? The authors address this issue with respect to the relation between metacognitive monitoring and metacognitive control. When self-regulation is goal driven, monitoring effects control processes so that increased processing effort should enhance feelings of competence and feelings of knowing. In contrast, when self-regulation is data driven, such feelings may be based themselves on the feedback from control processes, in which case they should decrease with increasing effort. Evidence for both monitoring-based control and control-based monitoring occurring even in the same situation is presented. The results are discussed with regard to the issue of the cause-and-effect relation between subjective experience and behavior. PMID- 16478317 TI - Stuck in a rut: perseverative response tendencies and the neuroticism-distress relationship. AB - Clinical views of neuroticism-linked distress often make reference to the perseverative sorts of mental processes that reinforce such experiences. The goal of the present 7 studies, involving 488 undergraduate participants, was to directly examine such perseverative processes. Individual differences in response perseveration were operationalized in terms of choice reaction time difficulties switching (vs. repeating) responses across consecutive trials. Response perseveration interacted with neuroticism in predicting negative emotion, dissatisfaction with life, and displays of negative emotion (Studies 1-4). Specifically, neuroticism-outcome relations were quite a bit stronger at high levels of perseveration. Additional studies (Studies 5-7) provided support for the convergent and discriminant validity of response perseveration. Overall, the results highlight the manner in which response perseveration reinforces experiences of negative emotion. PMID- 16478316 TI - Linking automatic evaluation to mood and information processing style: consequences for experienced affect, impression formation, and stereotyping. AB - According to the feelings-as-information account, a person's mood state signals to him or her the valence of the current environment (N. Schwarz & G. Clore, 1983). However, the ways in which the environment automatically influences mood in the first place remain to be explored. The authors propose that one mechanism by which the environment influences affect is automatic evaluation, the nonconscious evaluation of environmental stimuli as good or bad. A first experiment demonstrated that repeated brief exposure to positive or negative stimuli (which leads to automatic evaluation) induces a corresponding mood in participants. In 3 additional studies, the authors showed that automatic evaluation affects information processing style. Experiment 4 showed that participants' mood mediates the effect of valenced brief primes on information processing. PMID- 16478318 TI - Reasoning rats: forward blocking in Pavlovian animal conditioning is sensitive to constraints of causal inference. AB - Forward blocking is one of the best-documented phenomena in Pavlovian animal conditioning. According to contemporary associative learning theories, forward blocking arises directly from the hardwired basic learning rules that govern the acquisition or expression of associations. Contrary to this view, here the authors demonstrate that blocking in rats is flexible and sensitive to constraints of causal inference, such as violation of additivity and ceiling considerations. This suggests that complex cognitive processes akin to causal inferential reasoning are involved in a well-established Pavlovian animal conditioning phenomenon commonly attributed to the operation of basic associative processes. PMID- 16478319 TI - Interruption of the Tower of London task: support for a goal-activation approach. AB - Unexpected interruptions introduced during the execution phase of simple Tower of London problems incurred a time cost when the interrupted goal was retrieved, and this cost was exacerbated the longer the goal was suspended. Furthermore, time taken to retrieve goals was greater following a more complex interruption, indicating the processing limitations may be as important as time-based limitations in determining the ease of goal retrieval. Such findings cannot simply be attributed to task-switching costs and are evaluated in relation to current models of goal memory (E. M. Altmann & G. J. Trafton, 2002; J. R. Anderson & S. Douglass, 2001), which provide a useful basis for the investigation and interpretation of interruption effects. PMID- 16478320 TI - Perceptual encoding efficiency in visual search. AB - The authors present 10 experiments that challenge some central assumptions of the dominant theories of visual search. Their results reveal that the complexity (or redundancy) of nontarget items is a crucial but overlooked determinant of search efficiency. The authors offer a new theoretical outline that emphasizes the importance of nontarget encoding efficiency, and they test this proposal using dot pattern stimuli adapted from W. R. Garner and D. E. Clement (1963). The results provide converging support for the importance of nontarget encoding efficiency in accounting for visual search performance. PMID- 16478322 TI - Higher order pattern structure influences auditory representational momentum. AB - Representational momentum refers to the phenomenon that observers tend to incorrectly remember an event undergoing real or implied motion as shifted beyond its actual final position. This has been demonstrated in both visual and auditory domains. In 5 pitch discrimination experiments, listeners heard tone sequences that implied either linear, periodic, or null motions in pitch space. Their task was to judge whether the pitch of a probe tone following each sequence was the same or different from the final sequence tone. Results suggested that listeners made errors consistent with extrapolation of coherent pitch patterns (linear, periodic) but not with incoherent (null) ones. Hypotheses associated with internalized physical principles and pattern-based expectations are discussed. PMID- 16478323 TI - Syllables as processing units in handwriting production. AB - This research focused on the syllable as a processing unit in handwriting. Participants wrote, in uppercase letters, words that had been visually presented. The interletter intervals provide information on the timing of motor production. In Experiment 1, French participants wrote words that shared the initial letters but had different syllable boundaries. In Experiment 2, French- and Spanish speaking participants wrote cognates and pseudowords with a letter sequence that was always intrasyllabic in French and intersyllabic in Spanish. In Experiment 3, French-Spanish bilinguals wrote the cognates and pseudowords with the same type of sequences. In the 3 experiments, the critical interletter intervals were longer between syllables than within syllables, indicating that word syllable structure constrains motor production both in French and Spanish. PMID- 16478324 TI - Numbers and space: a computational model of the SNARC effect. AB - The SNARC (spatial numerical associations of response codes) effect reflects the tendency to respond faster with the left hand to relatively small numbers and with the right hand to relatively large numbers (S. Dehaene, S. Bossini, & P. Giraux, 1993). Using computational modeling, the present article aims to provide a framework for conceptualizing the SNARC effect. In line with models of spatial stimulus-response congruency, the authors modeled the SNARC effect as the result of parallel activation of preexisting links between magnitude and spatial representation and short-term links created on the basis of task instructions. This basic dual-route model simulated all characteristics associated with the SNARC effect. In addition, 2 experiments tested and confirmed new predictions derived from the model. PMID- 16478325 TI - Performance-related activity in medial rostral prefrontal cortex (area 10) during low-demand tasks. AB - Neuroimaging studies have frequently observed relatively high activity in medial rostral prefrontal cortex (PFC) during rest or baseline conditions. Some accounts have attributed this high activity to the occurrence of unconstrained stimulus independent and task-unrelated thought processes during baseline conditions. Here, the authors investigated the alternative possibility that medial rostral PFC supports attention toward the external environment during low-demand conditions. Participants performed a baseline simple reaction time (RT) task, along with 3 other tasks that differed in the requirement to attend to external stimuli versus stimulus-independent thought. Medial rostral PFC activation was observed in the baseline task and in a condition requiring strong engagement with external stimuli, relative to 2 conditions with a greater requirement for stimulus-independent thought. An important finding was that activity in this region was associated with faster RTs in the baseline task, ruling out an explanation in terms of task-unrelated thought processes during this condition. Thus, at least under certain circumstances, medial rostral PFC appears to support attention toward the external environment, facilitating performance in situations that do not require extensive processing of experimental stimuli. PMID- 16478326 TI - When is search for a static target among dynamic distractors efficient? AB - Intuitively, dynamic visual stimuli, such as moving objects or flashing lights, attract attention. Visual search tasks have revealed that dynamic targets among static distractors can indeed efficiently guide attention. The present study shows that the reverse case, a static target among dynamic distractors, allows for relatively efficient selection in certain but not all cases. A static target was relatively efficiently found among distractors that featured apparent motion, corroborating earlier findings. The important new finding was that static targets were equally easily found among distractors that blinked on and off continuously, even when each individual item blinked at a random rate. However, search for a static target was less efficient when distractors abruptly varied in luminance but did not completely disappear. The authors suggest that the division into the parvocellular pathway dealing with static visual information, on the one hand, and the magnocellular pathway common to motion and new object onset detection, on the other hand, allows for efficient filtering of dynamic and static information. PMID- 16478327 TI - Turning configural processing upside down: part and whole body postures. AB - Like faces, body postures are susceptible to an inversion effect in untrained viewers. The inversion effect may be indicative of configural processing, but what kind of configural processing is used for the recognition of body postures must be specified. The information available in the body stimulus was manipulated. The presence and magnitude of inversion effects were compared for body parts, scrambled bodies, and body halves relative to whole bodies and to corresponding conditions for faces and houses. Results suggest that configural body posture recognition relies on the structural hierarchy of body parts, not the parts themselves or a complete template match. Configural recognition of body postures based on information about the structural hierarchy of parts defines an important point on the configural processing continuum, between recognition based on first-order spatial relations and recognition based on holistic undifferentiated template matching. PMID- 16478328 TI - Judgments of path, not heading, guide locomotion. AB - To steer a course through the world, people are almost entirely dependent on visual information, of which a key component is optic flow. In many models of locomotion, heading is described as the fundamental control variable; however, it has also been shown that fixating points along or near one's future path could be the basis of an efficient control solution. Here, the authors aim to establish how well observers can pinpoint instantaneous heading and path, by measuring their accuracy when looking at these features while traveling along straight and curved paths. The results showed that observers could identify both heading and path accurately (approximately 3 degrees ) when traveling along straight paths, but on curved paths they were more accurate at identifying a point on their future path (approximately 5 degrees ) than indicating their instantaneous heading (approximately 13 degrees ). Furthermore, whereas participants could track changes in the tightness of their path, they were unable to accurately track the rate of change of heading. In light of these results, the authors suggest it is unlikely that heading is primarily used by the visual system to support active steering. PMID- 16478329 TI - The influence of linguistic experience on the cognitive processing of pitch in speech and nonspeech sounds. AB - In the present experiment, the authors tested Mandarin and English listeners on a range of auditory tasks to investigate whether long-term linguistic experience influences the cognitive processing of nonspeech sounds. As expected, Mandarin listeners identified Mandarin tones significantly more accurately than English listeners; however, performance did not differ across the listener groups on a pitch discrimination task requiring fine-grained discrimination of simple nonspeech sounds. The crucial finding was that cross-language differences emerged on a nonspeech pitch contour identification task: The Mandarin listeners more often misidentified flat and falling pitch contours than the English listeners in a manner that could be related to specific features of the sound structure of Mandarin, which suggests that the effect of linguistic experience extends to nonspeech processing under certain stimulus and task conditions. PMID- 16478330 TI - Frequency effects in spoken and visual word recognition: evidence from dual-task methodologies. AB - The authors report 3 dual-task experiments concerning the locus of frequency effects in word recognition. In all experiments, Task 1 entailed a simple perceptual choice and Task 2 involved lexical decision. In Experiment 1, an underadditive effect of word frequency arose for spoken words. Experiment 2 also showed underadditivity for visual lexical decision. It was concluded that word frequency exerts an influence prior to any dual-task bottleneck. A related finding in similar dual-task experiments is Task 2 response postponement at short stimulus onset asynchronies. This was explored in Experiment 3, and it was shown that response postponement was equivalent for both spoken and visual word recognition. These results imply that frequency-sensitive processes operate early and automatically. PMID- 16478331 TI - Representational specificity of within-category phonetic variation in the long term mental lexicon. AB - This study examines the potential encoding in long-term memory of subphonemic, within-category variation in voice onset time (VOT) and the degree to which this encoding of subtle variation is mediated by lexical competition. In 4 long-term repetition-priming experiments, magnitude of priming was examined as a function of variation in VOT in words with voiced counterparts (cape-gape) and without (cow-*gow) and words whose counterparts were high frequency (pest-best) or low frequency (pile-bile). The results showed that within-category variation was indeed encoded in memory and could have demonstrable effects on priming. However, there were also robust effects of prototypical representations on priming. Encoding of within-category variation was also affected by the presence of lexical counterparts and by the frequency of counterparts. PMID- 16478332 TI - The generalized optic acceleration cancellation theory of catching. AB - The generalized optic acceleration cancellation (GOAC) theory of catching proposes that the path of a fielder running to catch a ball is determined by the attempt to satisfy 2 independent constraints. The 1st is to keep the angle of elevation of gaze to the ball increasing at a decreasing rate. The 2nd is to control the rate of horizontal rotation necessary to maintain fixation on the ball. Depending on the lateral velocity of the ball relative to the fielder, this rate may be zero or constant at a negative or positive value. The authors show that a simulated fielder implementing the GOAC strategy follows a path indistinguishable from that of real fielders running to catch balls thrown on the same trajectories. PMID- 16478333 TI - Backward response-level crosstalk in the psychological refractory period paradigm. AB - Bottleneck models of psychological refractory period (PRP) tasks suggest that a Task 1 response should be unaffected by the Task 2 response in the same trial, because selection of the former finishes before selection of the latter begins. Contrary to this conception, the authors found backward response-level crosstalk effects in which Task 2 response force requirements influenced the force-time dynamics of Task 1 responses. Specifically, Task 2 required a hard or soft keypress response. Task 1 responses were harder when the upcoming Task 2 response was to press hard rather than soft, suggesting some activation of Task 2 response parameters before Task 1 processing reached the final ballistic motor output stage. A 3rd experiment using a flankers paradigm showed that this effect did not arise from automatic activation of responses by the stimuli associated with hard and soft responses. This backward response-level crosstalk extends previous findings of backward crosstalk in the PRP paradigm by showing that crosstalk can affect motor output as well as response time and can arise even when the 2 tasks being performed are not semantically related. PMID- 16478334 TI - Hands up: attentional prioritization of space near the hand. AB - This study explored whether hand location affected spatial attention. The authors used a visual covert-orienting paradigm to examine whether spatial attention mechanisms--location prioritization and shifting attention--were supported by bimodal, hand-centered representations of space. Placing 1 hand next to a target location, participants detected visual targets following highly predictive visual cues. There was no a priori reason for the hand to influence task performance unless hand presence influenced attention. Results showed that target detection near the hand was facilitated relative to detection away from the hand, regardless of cue validity. Similar facilitation was found with only proprioceptive or visual hand location information but not with arbitrary visual anchors or distant targets. Hand presence affected attentional prioritization of space, not the shifting of attention. PMID- 16478335 TI - The effect of voice onset time differences on lexical access in Dutch. AB - Effects on spoken-word recognition of prevoicing differences in Dutch initial voiced plosives were examined. In 2 cross-modal identity-priming experiments, participants heard prime words and nonwords beginning with voiced plosives with 12, 6, or 0 periods of prevoicing or matched items beginning with voiceless plosives and made lexical decisions to visual tokens of those items. Six-period primes had the same effect as 12-period primes. Zero-period primes had a different effect, but only when their voiceless counterparts were real words. Listeners could nevertheless discriminate the 6-period primes from the 12- and 0 period primes. Phonetic detail appears to influence lexical access only to the extent that it is useful: In Dutch, presence versus absence of prevoicing is more informative than amount of prevoicing. PMID- 16478336 TI - Eye movements to pictures reveal transient semantic activation during spoken word recognition. AB - Two experiments explore the activation of semantic information during spoken word recognition. Experiment 1 shows that as the name of an object unfolds (e.g., lock), eye movements are drawn to pictorial representations of both the named object and semantically related objects (e.g., key). Experiment 2 shows that objects semantically related to an uttered word's onset competitors become active enough to draw visual attention (e.g., if the uttered word is logs, participants fixate on key because of partial activation of lock), despite that the onset competitor itself is not present in the visual display. Together, these experiments provide detailed information about the activation of semantic information associated with a spoken word and its phonological competitors and demonstrate that transient semantic activation is sufficient to impact visual attention. PMID- 16478337 TI - Consequences of lexical stress on learning an artificial lexicon. AB - Four experiments examined effects of lexical stress on lexical access for recently learned words. Participants learned artificial lexicons (48 words) containing phonologically similar items and were tested on their knowledge in a 4 alternative forced-choice (4AFC) referent-selection task. Lexical stress differences did not reduce confusions between cohort items: KAdazu and kaDAzeI were confused with one another in a 4AFC task and in gaze fixations as often as BOsapeI and BOsapaI. However, lexical stress did affect the relative likelihood of stress-initial confusions when words were embedded in running nonsense speech. Words with medial stress, regardless of initial vowel quality, were more prone to confusions than words with initial stress. The authors concluded that non-initial stress, particularly when wor segmentation is difficult, may serve as "noise" that alters lexical learning and lexical access. PMID- 16478338 TI - Part-list cuing can be transient and lasting: the role of encoding. AB - The presentation of a subset of learned items as retrieval cues can have detrimental effects on recall of the remaining items. For 2 types of encoding conditions, the authors examined in 3 experiments whether such part-list cuing is a transient or a lasting phenomenon. Across the experiments, the detrimental effect of part-list cues was consistently found to be transient with a high degree of interim associations and lasting with a low degree. These results indicate that the persistence of part-list cuing depends on encoding, thus challenging both strategy disruption and retrieval inhibition as general accounts of part-list cuing. A 2-mechanism account is provided according to which the 2 mechanisms mediate the effect in different encoding conditions. PMID- 16478339 TI - Episodic inhibition. AB - Six experiments examined the proposal that an item of long-term knowledge can be simultaneously inhibited and activated. In 2 directed forgetting experiments items to-be-forgotten were found to be inhibited in list-cued recall but activated in lexical decision tasks. In 3 retrieval practice experiments, unpracticed items from practiced categories were found to be inhibited in category-cued recall but were primed in lexical decision. If, however, the primes and targets in lexical decision were taken directly from the study list, inhibition was observed. Finally, it was found that when items highly associated with a study list were processed in between study and test, no inhibition in recall was present. These, and a broad range of other findings, can be explained by the concept of "episodic inhibition," which proposes that episodic memories retain copies of semantic knowledge structures that preserve patterns of activation/inhibition originally generated in those structures during encoding. ((c) 2006 APA, all rights reserved). PMID- 16478340 TI - Scene and position specificity in visual memory for objects. AB - This study investigated whether and how visual representations of individual objects are bound in memory to scene context. Participants viewed a series of naturalistic scenes, and memory for the visual form of a target object in each scene was examined in a 2-alternative forced-choice test, with the distractor object either a different object token or the target object rotated in depth. In Experiments 1 and 2, object memory performance was more accurate when the test object alternatives were displayed within the original scene than when they were displayed in isolation, demonstrating object-to-scene binding. Experiment 3 tested the hypothesis that episodic scene representations are formed through the binding of object representations to scene locations. Consistent with this hypothesis, memory performance was more accurate when the test alternatives were displayed within the scene at the same position originally occupied by the target than when they were displayed at a different position. PMID- 16478341 TI - Conjunction errors, recollection-based rejections, and forgetting in a continuous recognition task. AB - Six experiments investigated conjunction memory errors (e.g., falsely remembering blackbird after studying parent words blackmail and jailbird) in a continuous recognition procedure with a parent-conjunction lag manipulation. In 4 experiments (1A, 1B, 2A, and 2B) "recollect" judgments, which indexed recall of parent words, showed that participants can use recollection to prevent conjunction errors. "Recollect" judgments, as well as overt recall of parent words (in Experiments 2A and 2B), dropped sharply from a lag of 0 to 1 word, then stabilized from a lag of 1 to 20 words. Thus, the "recollect" responses and overt recall demonstrate a step function of forgetting over short intervals. These data generalized to cued recall in Experiments 3A and 3B with the first morpheme (e.g., black) as the cue, though recall conjunction errors occurred infrequently relative to recognition conjunction errors. Overall, the results support the idea that automatic and controlled processes contribute to memory performance. PMID- 16478342 TI - Does emotion help or hinder immediate memory? Arousal versus priority-binding mechanisms. AB - People recall taboo words better than neutral words in many experimental contexts. The present rapid serial visual presentation (RSVP) experiments demonstrated this taboo-superiority effect for immediate recall of mixed lists containing taboo and neutral words matched for familiarity, length, and category coherence. Under binding theory (MacKay et al., 2004), taboo superiority reflects an interference effect: Because the emotional reaction system prioritizes binding mechanisms for linking the source of an emotion to its context, taboo words capture the mechanisms for encoding list context in mixed lists, impairing the encoding of adjacent neutral words when RSVP rates are sufficiently rapid. However, for pure or unmixed lists, binding theory predicted no better recall of taboo-only than of neutral-only lists at fast or slow rates. Present results supported this prediction, suggesting that taboo superiority in immediate recall reflects context-specific binding processes, rather than context-free arousal effects, or emotion-linked differences in rehearsal, processing time, output interference, time-based decay, or guessing biases. PMID- 16478343 TI - Binding, relational memory, and recall of naturalistic events: a developmental perspective. AB - This research was an investigation of children's performance on a task that requires memory binding. In Experiments 1 and 2, 4-year-olds, 6-year-olds, and adults viewed complex pictures and were tested on memory for isolated parts in the pictures and on the part combinations (combination condition). The results suggested improvement in memory for the combinations between the ages of 4 and 6 years but not in memory for the isolated parts. In Experiments 2 and 3, the authors also examined the developmental relationship between performance in the combination condition and free recall of a naturalistic event, finding preliminary evidence that performance on a memory task that requires binding is positively related to performance in episodic memory. PMID- 16478345 TI - Interactions between encoding and retrieval in the domain of sequence-learning. AB - In this article, the authors propose to characterize sequence learning in terms of automatic versus non-automatic processing and to apply this contrast independently to knowledge acquisition and retrieval. In several experiments of sequence learning, automaticity of both the acquisition and retrieval of the acquired knowledge was independently assessed. It was found that the sequence learning order can be demonstrated under all combinations of knowledge acquisition and retrieval. In particular, at least in the simple sequences the authors used, this applies in cases in which both the acquisition and the retrieval of knowledge are strictly automatic--that is, when neither is required for the task nor beneficial to deliberate behavior. The proposed framework has implications for the notion of sequence learning and the investigation of learning in general. PMID- 16478346 TI - Game relativity: how context influences strategic decision making. AB - Existing models of strategic decision making typically assume that only the attributes of the currently played game need be considered when reaching a decision. The results presented in this article demonstrate that the so-called "co-operativeness" of the previously played prisoner's dilemma games influence choices and predictions in the current prisoner's dilemma game, which suggests that games are not considered independently. These effects involved reinforcement based assimilation to the previous choices and also a perceptual contrast of the present game with preceding games, depending on the range and the rank of their co-operativeness. A. Parducci's (1965) range frequency theory and H. Helson's (1964) adaptation level theory are plausible theories of relative judgment of magnitude information, which could provide an account of these context effects. PMID- 16478344 TI - A diffusion model analysis of adult age differences in episodic and semantic long term memory retrieval. AB - Two experiments investigated adult age differences in episodic and semantic long term memory tasks, as a test of the hypothesis of specific age-related decline in context memory. Older adults were slower and exhibited lower episodic accuracy than younger adults. Fits of the diffusion model (R. Ratcliff, 1978) revealed age related increases in non-decisional reaction time for both episodic and semantic retrieval. In Experiment 2, an age difference in boundary separation also indicated an age-related increase in conservative criterion setting. For episodic old-new recognition (Experiment 1) and source memory (Experiment 2), there was an age-related decrease in the quality of decision-driving information (drift rate). As predicted by the context-memory deficit hypothesis, there was no corresponding age-related decline in semantic drift rate. PMID- 16478347 TI - Recognition is used as one cue among others in judgment and decision making. AB - Three experiments with paired comparisons were conducted to test the non compensatory character of the recognition heuristic (D. G. Goldstein & G. Gigerenzer, 2002) in judgment and decision making. Recognition and knowledge about the recognized alternative were manipulated. In Experiment 1, participants were presented pairs of animal names where the task was to select the animal with the larger population. In Experiment 2, participants chose the safer 1 out of 2 airlines, and 3 knowledge cues were varied simultaneously. Recognition effects were partly compensated by task-relevant knowledge. The compensatory effects were additive. Decisions were slower when recognition and knowledge were incongruent. In Experiment 3, compensatory effects of knowledge and recognition were found for the city-size task which had originally been used to demonstrate the non compensatory character of the recognition heuristic. These results suggest that recognition information is not used in an all-or-none fashion but is integrated with other types of knowledge in judgment and decision making. PMID- 16478348 TI - On the role of causal intervention in multiple-cue judgment: positive and negative effects on learning. AB - Previous studies have suggested better learning when people actively intervene rather than when they passively observe the stimuli in a judgment task. In 4 experiments, the authors investigated the hypothesis that this improvement is associated with a shift from exemplar memory to cue abstraction. In a multiple cue judgment task with continuous cues, the data replicated the improvement with intervention and participants who experimented more actively produced more accurate judgments. In a multiple-cue judgment task with binary cues, intervention produced poorer accuracy and participants who experimented more actively produced poorer judgments. These results provide no support for a representational shift but suggest that the improvement with active intervention may be limited to certain tasks and environments. PMID- 16478350 TI - Coping responses of Asian, Black, and Latino/Latina New York City residents following the September 11, 2001 terrorist attacks against the United States. AB - This study examined mechanisms for coping with adversity in a sample of 24 Asian, Black, and Latino/Latina residents of New York City following the September 11, 2001 World Trade Center (WTC) terrorist attacks. Using consensual qualitative research methodology (C. E. Hill, B. J. Thompson, & E. N. Williams, 1997), the authors identified 7 broad coping domains used by the participants: (a) sought additional information about the WTC tragedy, (b) expressed a range of emotions, (c) sought or gave support, (d) engaged in religious or spiritual activities, (e) avoidance, (f) forbearance, and (g) used indigenous healing techniques. Although there were similarities across racial or cultural groups and genders with regard to the coping responses used, there also were unique coping strategies by racial or cultural background and gender. PMID- 16478351 TI - Adolescent alcohol use and suicide indicators among adolescents in Hawaii. AB - Research on suicide has focused on gender, age, ethnicity, and psychiatric profiles. However, few studies have examined alcohol use and its relationship to suicide among Native Hawaiians and other Asian American/Pacific Islanders. This study analyzes data from the 1997 and 1999 Hawaii Youth Risk Behavior Survey to examine whether alcohol problems increase the risk for suicide indicators (as evidenced by responses to questions asking whether an individual has considered, planned, attempted, or required treatment for a suicide attempt). Drinking pattern was the best predictor for all suicide indicators. School and community based programs can help to increase an adolescent's knowledge about the consequences of alcohol use and prevention of suicide. PMID- 16478352 TI - Self-concept in Mexican American girls and boys: validating the Self-Description Questionnaire-I. AB - This study examined the factor structure and psychometric properties of the Self Description Questionnaire-I (SDQ-I) in a sample of Mexican American children. Findings provide support for both the classic multidimensional and hierarchical SDQ factor structure and a nonhierarchical model that incorporates a dimension not included in the classic model (i.e., General-Self, a global measure of self concept). Results of a multiple-group confirmatory factor analysis demonstrate the nonhierarchical model's overall invariance across gender. Gender differences were found, however, on mean SDQ-I subscale scores: Boys reported higher self concept than girls on the Physical Abilities and Physical Appearance subscales, but girls had higher self-concept than boys on the Reading subscale. PMID- 16478353 TI - Culture-based perceptions of academic achievement among low-income elementary students. AB - This study examined the influence of culture on students' perceptions of academic success. Students read scenarios depicting hypothetical classmates achieving success through the cultural themes of individualism, competition, communalism, or verve. Students reported their social endorsement for the hypothetical classmates. A 2x4 repeated measures analysis, examining the effects of cultural group and cultural theme on students' endorsement, revealed an interaction between the two variables. African American students were significantly more accepting of communal and vervistic high-achieving peers than European American students. European American students endorsed individualistic and competitive high achievers significantly more than African American students. These and other findings suggest that the value students attach to academic success should not be understood in the absence of cultural considerations. PMID- 16478354 TI - Clinician race, situational attributions, and diagnoses of mood versus schizophrenia disorders. AB - This research examined clinicians' consideration of situational factors in diagnostic decisions of mood vs. schizophrenia disorders among psychiatric inpatients from a low-income, African American community. Clinicians completed questionnaires describing their diagnostic decisions. Responses reflecting the usage of situational information were investigated. African American clinicians used situational information more than non-African American clinicians. However, this increased attention to situational information was not uniquely associated with a particular diagnostic decision for African American clinicians. In contrast, consideration of situational attributions by non-African American clinicians did increase the probability of a mood diagnosis. Logistic regression analyses suggested differential application of a diagnostic standard among African American and non-African American clinicians. Implications for enhancing the cultural sensitivity of diagnosis practices are discussed. PMID- 16478355 TI - Child-care effect sizes for the NICHD Study of Early Child Care and Youth Development. AB - This report summarizes findings from the National Institute of Child Health and Human Development Study of Early Child Care and Youth Development as effect sizes for exclusive maternal care and--for children in child care--type, quality, and quantity of care. Children (n = 1,261) were recruited at birth and assessed at 15, 24, 36, and 54 months. Exclusive maternal care did not predict child outcomes, but multiple features of child-care experience were modestly to moderately predictive. Higher quality child care was related to advanced cognitive, language, and preacademic outcomes at every age and better socioemotional and peer outcomes at some ages. More child-care hours predicted more behavior problems and conflict, according to care providers. More center care time was related to higher cognitive and language scores and more problem and fewer prosocial behaviors, according to care providers. Child-care effect sizes are discussed from 3 perspectives: (a) absolute effect sizes, reflecting established guidelines; (b) relative effect sizes, comparing child-care and parenting effects; and (c) possible individual and collective implications for the large numbers of children experiencing child care. PMID- 16478356 TI - Minority students of color and the psychology graduate pipeline: disquieting and encouraging trends, 1989-2003. AB - Trends since 1989 in the minority graduate pipeline in psychology are examined, with special focus on trends in recent years. Encouraging trends generally outweigh troubling ones at lower levels of the pipeline. However, in recent years disquieting trends dominate at the higher pipeline levels. Promising trends include a rise in the percentage (to nearly 25%) of minority psychology students receiving the bachelor's degree and a rise to more than 20% receiving the master's degree. Troubling trends include the stalling of growth in minority PhD degree receipt since 1999 and the lack of growth in the percentage of African American and Hispanic/Latino(a) students entering PhD departments. Given the mixed findings, one of the highest priorities for psychology must be continued and persistent efforts to develop practices and policies that enhance recruitment, high levels of achievement, and degree receipt for students of color. PMID- 16478358 TI - Exemplary efforts in psychology to recruit and retain graduate students of color. AB - Many psychology departments are striving for a greater representation of students of color within their graduate preparation programs with the aim of producing a more diverse pool of psychological service providers, scientists, and educators. To help improve the minority pipeline in psychology, the authors identify and describe recruitment and retention strategies used at 11 departments and programs considered to be making exemplary efforts to attract and retain minority students of color. The strategies most consistently used included engaging current minority faculty and students in recruitment activities, offering attractive financial aid packages, having faculty members make personal contacts with prospective students, creating linkages with historical institutions of color, having (or approached having) a critical mass of faculty and students of color, offering a diversity issues course, and engaging students in diversity issues research. Despite the similarities, the programs and departments were each distinctive and innovative in their overall approaches to student recruitment and retention. Highlighting the strategies used at successful institutions may help others develop plans for improving the minority pipeline within their own departments and programs. PMID- 16478357 TI - Increasing the number of psychologists of color: public policy issues for affirmative diversity. AB - This article identifies the key issues involved in the debate about affirmative action. The June 2003 Supreme Court decisions allowing consideration of race to ensure that there is a "critical mass" of African American, Latino/Latina, and Native American applicants to higher education are addressed. Social psychologists have identified key myths and provided clarifications about the need for and consequences of strategies used to promote equal opportunity for persons of color and women. A brief history of affirmative action and of the problems it was designed to solve is provided. The accomplishments, benefits, and compelling interest of diversity and affirmative action are described, as well as the concerns and counterpoints. The lack of a substantial applicant pool in psychology hinders progress toward diversity. Alternative strategies for remedying this lack beyond affirmative admissions policies in psychology are briefly discussed. PMID- 16478359 TI - Personal reflections: barriers and strategies in increasing diversity in psychology. AB - In this article, six faculty and students of color who participated in a panel discussion at a symposium during the National Multicultural Conference and Summit of 2003 talk about the barriers they encountered and continue to encounter in their graduate training and places of employment. They also discuss strategies they found to be effective, enhancing, and positive and suggest other possibilities. The contributors describe their relationships with dominant-group and minority peers and talk about how issues of social class, disability, and sexual orientation as well as color have been part of their experience. PMID- 16478360 TI - Urie Bronfenbrenner (1917-2005). AB - Presents an obituary for Urie Bronfenbrenner (1917-2005). Before Urie Bronfenbrenner, child psychologists studied the child, sociologists the family, anthropologists the society, economists the economic framework, and political scientists the structure. As the result of Urie's extension of the concept of the ecology of human development, these environments--from the family to economic and political structures--are viewed as part of the life course, embracing both childhood and adulthood. Bronfenbrenner, widely regarded as one of the world's leading scholars in developmental psychology, child rearing, and human ecology- the interdisciplinary domain he helped popularize--died at his home in Ithaca, New York, on September 25, 2005, at the age of 88. He was the Jacob Gould Sherman Professor Emeritus of Human Development and of Psychology at Cornell University, where he spent most of his professional career. A brief biography of Bronfenbrenner is followed by an overview of his published work, his theories and other influential accomplishments. PMID- 16478361 TI - Theodore X. Barber (1927-2005). AB - Presents an obituary for Theodore Xenophon Barber (1927-2005), one of the most prolific and influential researchers in the field of hypnosis. At the time of his death he was an active scholar in his private research enterprise, the Interdisciplinary Science Research Institute. A brief biography of Barber is followed by an overview of his published work, his theories and other influential accomplishments. Although hypnosis was the main focus of Barber's research, his interests and research encompassed other topics, including the phenomenon of investigator bias, psychical phenomena, and even comparative psychology. PMID- 16478362 TI - Is the evidence on ethnicity and intelligence conclusive? PMID- 16478364 TI - Examining unproven assumptions of Galton's nature-nurture paradigm. PMID- 16478366 TI - On the complexity of race. PMID- 16478363 TI - Race--social, biological, or lemonade? PMID- 16478368 TI - Managing oncology costs. AB - This monograph will review the burden of illness in oncology, suggest a framework for evaluating oncology costs and consequences, identify economic modeling formats in cancer care, and explore methods of cost control for cancer care. PMID- 16478369 TI - Electromyographic study of motor learning for a voice production task. AB - PURPOSE: This study's broad objective was to examine the effectiveness of surface electromyographic (EMG) biofeedback for motor learning in the voice production domain. The specific objective was to examine whether concurrent or terminal biofeedback would facilitate learning for a relaxed laryngeal musculature task during spoken reading. METHOD: Twenty-two healthy adult speakers were randomly assigned to 1 of 2 groups. One group received real-time EMG waveform displays of muscle activation from bilateral thyrohyoid sites during reading trials (concurrent feedback group). The other group received static terminal EMG waveform displays about activation levels for the same sites on completion of successive trials (terminal feedback group). All participants were instructed to minimize EMG amplitudes from the thyrohyoid sites during phonation in an oral reading task. Signals were also collected from control, orofacial sites, but participants received neither instructions nor feedback for those sites. RESULTS: The pooled data (2 feedback groups x 2 electrode sites) showed that, overall, muscle activation levels did decrease across baseline, training, and no-feedback test phases. However, no clear evidence was seen of reliable changes in the targeted laryngeal muscle activation levels across the phases, for either the concurrent or the terminal feedback groups. Paradoxically, and entirely unanticipated, reliable decreases were seen in muscle activation for the orofacial, no-feedback control sites. Those decreases were equivalent across concurrent and terminal feedback groups. CONCLUSIONS: The unanticipated findings indicate that the provision of biofeedback for a target muscle group facilitated incidental learning in another, untargeted muscle group. Discussion focuses on the possible role of locus of attention in motor learning. Building on literature from other domains, the hypothesis is advanced that attention to muscular contractile force during training trials may suppress intentional learning for attended target sites but may benefit incidental learning for nearby, unattended sites. PMID- 16478370 TI - Comparison of clinician judgments and measurements of swallow response time: a preliminary report. AB - Practicing clinicians frequently offer judgments about aspects of swallowing physiology rather than performing actual measurements. Little is known about the accuracy of those judgments. The purpose of this preliminary study was to explore agreement of clinicians' judgments of pharyngeal swallow response time (PSRT) with temporal measurements of PSRT. In preparation for a larger study, PSRT was measured from the first 3 ml liquid bolus swallow that appeared in each of 20 videofluorographic swallowing evaluations. The same 20 swallows were then shown to 3 clinicians who were instructed to subjectively rate PSRT. The reliability of the PSRT measurements was strong (r > .95). Intrajudge and interjudge agreement was better than chance in all but 1 interjudge comparison. Percentage agreement between clinicians' judgments and the measurements, when the measurements were categorized as either not delayed or delayed, ranged from 60% to 95%. Chi-square and Phi statistics comparing the outcomes of clinicians' ratings with the measurement outcomes were significant, supporting the agreement of the judgments with the measurements. Although the results of tests of agreement were found to be acceptable, clinician experience and training remain important issues whenever clinical judgments are involved. Larger studies are needed to establish the accuracy and importance of clinicians' judgments of PSRT and observations of swallowing physiology. PMID- 16478372 TI - Breath-group intelligibility in dysarthria: characteristics and underlying correlates. AB - PURPOSE: This study was designed to determine whether within-speaker fluctuations in speech intelligibility occurred among speakers with dysarthria who produced a reading passage, and, if they did, whether selected linguistic and acoustic variables predicted the variations in speech intelligibility. METHOD: Participants with dysarthria included a total of 10 persons with Parkinson's disease and amyotrophic lateral sclerosis; a control group of 10 neurologically normal speakers was also studied. Each participant read a passage that was subsequently separated into consecutive breath groups for estimates of individual breath group intelligibility. Sixty listeners participated in 2 perceptual experiments, generating intelligibility scores across speakers and for each breath group produced by speakers with dysarthria. RESULTS: Individual participants with dysarthria had fluctuations in intelligibility across breath groups. Breath groups of participants with dysarthria had fewer average words and reduced interquartile ranges for the 2nd formant, the latter a global measure of articulatory mobility. Regression analyses with intelligibility measures as the criterion variable and linguistic and acoustic measures as predictor variables produced significant functions both within and across speakers, but the solutions were not the same. CONCLUSIONS: Linguistic or acoustic variables that predict across-speaker variations in speech intelligibility may not function in the same way when within-speaker variations in intelligibility are considered. PMID- 16478371 TI - Utility of clinical swallowing examination measures for detecting aspiration post stroke. AB - The purpose of this investigation was to determine the utility of clinical swallowing examination (CSE) measures for detecting aspiration as defined by videofluoroscopic swallowing examination (VFSE). This study, involving 165 participants, is a follow-up to a previously published investigation of 60 participants. Findings are compared with that investigation as well as with other research on CSEs. The results suggest that clinicians can make an accurate judgment of the occurrence of aspiration in most post-stroke patients. However, ruling out aspiration when it is absent appears more problematic. More work needs to be done if data collected from non-instrumented examinations are to be strongly predictive of the presence and absence of aspiration on VFSE. At present, there are no data to suggest that CSEs can be used to quantify aspiration or make adequate recommendations regarding patient care. PMID- 16478373 TI - Velopharyngeal port status during classical singing. AB - PURPOSE: This investigation was undertaken to examine the status of the velopharyngeal (VP) port during classical singing. METHOD: Using aeromechanical instrumentation, nasal airflow (mL/s), oral pressure (cm H2O), and VP orifice area estimates (cm2) were studied in 10 classically trained sopranos during singing and speaking. Each participant sang and spoke 3 nonsense words-/hampa/, /himpi/, and /humpu/-at 3 loudness levels (loud vs. comfortable vs. soft) and 3 pitches (high vs. comfortable vs. low), using a within-subject experimental design including all possible combinations. RESULTS: In general, nasal airflow, oral pressure, and VP area estimates were significantly greater for singing as compared to speech, and nasal airflow was observed during non-nasal sounds in all participants. Anticipatory nasal airflow was observed in 9 of 10 participants for singing and speaking and was significantly greater during the first vowel in /hampa/ versus /himpi/ and /humpu/. The effect of vowel height on nasal airflow was also significantly influenced by loudness and pitch. CONCLUSIONS: The results from this investigation indicate that at least some trained singers experience regular VP opening during classical singing. Vowel height seems to influence this effect. Future research should consider the effects of voice type, gender, experience level, performance ability, and singing style on VP valving in singers. PMID- 16478374 TI - Examination of strength training and detraining effects in expiratory muscles. AB - PURPOSE: The purpose of this study was to determine strength gains following expiratory muscle strength training (EMST) and to determine detraining effects when the training stimulus is removed. METHOD: Thirty-two healthy participants were enrolled in an EMST program. Sixteen participants trained for 4 weeks (Group 1) and 16 participants trained for 8 weeks (Group 2). All 32 participants were detrained for 8 weeks. Maximum expiratory pressure (MEP) was used to document change in expiratory muscle strength throughout the study. RESULTS: Group 1 had a 41% increase and Group 2 had a 51% increase in MEP following the training. Mean MEP, for both groups, was significantly greater than baseline at the end of the training period (p = .0001), at the 4th week of detraining (p = .0001), and at the 8th week of detraining (p = .0001). The results also indicated that there was no significant difference in mean MEP between the groups at baseline, end of training, or throughout the detraining period (p = .960). DISCUSSION: The results suggest that expiratory muscle strength gains following a 4- and 8-week EMST program do not differ significantly. Additionally, detraining rates do not appear to be dependent on length of training time. PMID- 16478376 TI - The onset of tense marking in children at risk for specific language impairment. AB - PURPOSE: The purpose of this study was to develop and evaluate measures reflecting the onset of tense marking for children between the ages of 2;0 (years;months) and 3;0. METHOD: The validity of 4 cumulative measures of tense marker emergence and productivity was evaluated relative to existing measures of early grammatical development in a sample of 20 children followed longitudinally. Fourteen children were at risk for specific language impairment (AR-SLI group), and 6 children had low average language abilities (LA group). RESULTS: All measures of onset were highly correlated with the traditional measures; however, children's progress toward mastery of grammatical tense marking was best explained by the productivity of their tense marking systems. Finally, the onset measures imposing productivity requirements best differentiated children in the LA group from those in the AR-SLI group. CONCLUSIONS: The clinical implications for using the late onset of tense marking to improve the early identification of SLI are discussed. PMID- 16478375 TI - Control of voice-onset time in the absence of hearing: a review. AB - The relation between partial or absent hearing and control of the voicing contrast has long been of interest to investigators, in part because speakers who are born deaf characteristically have great difficulty mastering the contrast and in part for the light it can cast on the role of hearing in the acquisition and maintenance of phonological contrasts in general. One of the phonetic characteristics that distinguish voiced from voiceless plosives in English (p/b, t/d, k/g) is voice onset time (VOT): the interval from plosive release to the onset of voicing of the following vowel. This article first reviews research on VOT anomalies in the speech production of prelingually and postlingually deaf speakers. Then it turns to studies of the mechanisms in speech breathing, phonation and articulation that underlie those anomalies. In both populations of speakers, there is a tendency for the difference between voiced and voiceless VOT to be reduced, to the point for many speakers that there is in effect a substitution of the voiced for the voiceless cognate. The separation of the cognate VOTs can be enhanced when some hearing is restored with a cochlear implant. Both populations also present anomalies in speech breathing that can hinder the development of intraoral pressures and transglottal pressure drops that are required for the production of the VOT contrast. Its successful management further requires critical timing among phonatory and articulatory gestures, most of which are not visible, rendering the VOT contrast a particular challenge in the absence of hearing. PMID- 16478377 TI - The development of expressive elaboration in fictional narratives. AB - PURPOSE: This study analyzed the development of expressive elaboration in fictional narratives for school-age children. METHOD: The analysis was derived from high-point analysis, but it was tailored to capture the artful aspects of fictional storytelling. Narratives were elicited with a short picture sequence of a likely life event from 293 children whose ages ranged from 5 to 12 years. RESULTS: Results showed a significant age effect for expressive elaboration with narrative length controlled. For three age clusters (5-6 years, 7-9 years, and 10 12 years), the 13 types of expressive elaboration showed diverse patterns of acquisition in terms of presence, frequency, and developmental change. Appendages (introducer, abstract, theme, coda, ender) were lowest in both presence and frequency, and increased in presence with age. Orientations (names, relations, personality) were more common and increased in presence with age. Evaluations (modifiers, expressions, repetition, internal states, dialogue) were most frequent and showed age changes in both presence and frequency. CLINICAL IMPLICATIONS: This study provides an additional window on narrative competence. The analysis and results can guide narrative assessment and intervention. PMID- 16478379 TI - Verb learning in children with SLI: frequency and spacing effects. AB - PURPOSE: This study explored the effect of frequency (number of presentations), and spacing (period between presentations) on verb learning in children with specific language impairment (SLI). Children learn words more efficiently when presentations are frequent and appropriately spaced, and this study investigated whether children with SLI likewise benefit. Given that these children demonstrate greater frequency dependence and rapid forgetting of recently acquired words, an investigation of frequency and spacing in this population is especially warranted. METHOD: Twenty-four children with SLI (mean age 5;6 [years;months]) and 24 language-matched control children (mean age 3;4) were taught novel verbs during play sessions. In a repeated measures design, 4 experimental conditions combined frequency (12 or 18 presentations) and spacing (all presentations in 1 session, or spread over 4 days). Comprehension and production probes were administered after the final session and 1 week later. RESULTS: Although the children with SLI benefited significantly from frequent and widely spaced presentations, there were no significant effect in the control group. The language-impaired children showed rapid forgetting. CONCLUSIONS: The frequency and spacing of presentations crucially affect the verb learning of children with SLI. A training regimen characterized by appropriately spaced intervals and moderate repetition will optimally benefit lexical learning. PMID- 16478380 TI - An application of Rasch analysis to the measurement of communicative functioning. AB - PURPOSE: The purposes of this investigation were to examine the construct dimensionality and range of ability effectively measured by 28 assessment items obtained from 3 different patient-reported scales of communicative functioning, and to provide a demonstration of how the Rasch approach to measurement may contribute to the definition of latent constructs and the development of instruments to measure them. METHOD: Item responses obtained from 421 stroke survivors with and without communication disorders were examined using the Rasch partial credit model. The dimensionality of the item pool was evaluated by (a) examining correlations of Rasch person ability scores obtained separately from each of the 3 scales, (b) iteratively excluding items exceeding mean square model fit criteria, and (c) using principal-components analysis of Rasch model residuals. The range of ability effectively measured by the item pool was examined by comparing item difficulty and category threshold calibrations to the distribution of person ability scores and by plotting the modeled standard error of person ability estimates as a function of person ability level. RESULTS: The results indicate that most assessment items fit a unidimensional measurement model, with the notable exception of items relating to the use of written communication. The results also suggest that the range of ability that could be reliably measured by the current item pool was restricted relative to the range of ability observed in the patient sample. CONCLUSIONS: It is concluded that (a) a mature understanding of communicative functioning as a measurement construct will require further research, (b) patients with stroke-related communication disorders will be better served by the development of instruments measuring a wider range of communicative functioning ability, and (c) the theoretical and methodological tools provided by the Rasch family of measurement models may be productively applied to these efforts. PMID- 16478378 TI - Are specific language impairment and dyslexia distinct disorders? AB - PURPOSE: The purpose of this study was to determine whether specific language impairment (SLI) and dyslexia are distinct developmental disorders. METHOD: Study 1 investigated the overlap between SLI identified in kindergarten and dyslexia identified in 2nd, 4th, or 8th grades in a representative sample of 527 children. Study 2 examined phonological processing in a subsample of participants, including 21 children with dyslexia only, 43 children with SLI only, 18 children with SLI and dyslexia, and 165 children with typical language/reading development. Measures of phonological awareness and nonword repetition were considered. RESULTS: Study 1 showed limited but statistically significant overlap between SLI and dyslexia. Study 2 found that children with dyslexia or a combination of dyslexia and SLI performed significantly less well on measures of phonological processing than did children with SLI only and those with typical development. Children with SLI only showed only mild deficits in phonological processing compared with typical children. CONCLUSIONS: These results support the view that SLI and dyslexia are distinct but potentially comorbid developmental language disorders. A deficit in phonological processing is closely associated with dyslexia but not with SLI when it occurs in the absence of dyslexia. PMID- 16478381 TI - Looking while listening and speaking: eye-to-face gaze in adolescents with and without traumatic brain injury. AB - PURPOSE: The purpose of this study was to address the lack of quantitative data on eye-to-face gaze (also known as eye contact) in the literature on pragmatic communication. The study focused on adolescents and young adults with traumatic brain injury (TBI), as gaze often is included in social skills intervention in this population. METHOD: Gaze times were calculated for participants with TBI (n = 16) and their typically developing (TD) peers (n = 16) engaged in 3-min extemporaneous conversations. RESULTS: The TD group members looked at the face of their conversation partner an average of 62% of the time while listening and 43% of the time while speaking, versus 67% and 51%, respectively, for the TBI group. There were no significant between-groups differences in average gaze times, but the within-group variability was significantly greater in the TBI group. IMPLICATIONS: As there was no evidence of a uniform trend in gaze times among participants with TBI, general intervention to increase eye contact does not appear warranted. Instead, goals must consider that gaze is a highly complex behavior, not necessarily indicative of attention to one's partner, and that there are potential reasons for gaze aversion in individuals with cognitive limitations. PMID- 16478382 TI - The processing of morphology in old age: evidence from Hebrew. AB - PURPOSE: Taking advantage of the rich morphological structure of Hebrew, the current article aims to examine whether age affects the processing of morphological forms through an investigation of 2 systematic morphological paradigms. METHOD: Forty-eight young and 48 old Hebrew speakers completed 2 experiments: the 1st investigated sensitivity to subject-verb gender incongruity in a reading task, and the 2nd examined parsing of pseudoverbs containing existing and nonexisting consonantal roots in a lexical-decision task. RESULTS: Older adults were slower relative to the young, but both groups were slower on incongruent relative to congruent targets and on a pseudoverb with a real root relative to a pseudoverb with a nonexistent root. In both experiments the interaction between condition and age was statistically significant. CONCLUSIONS: While older adults demonstrate preserved morphological parsing abilities, possible explanations for the interaction effect include cognitive slowing or deficient inhibitory control. PMID- 16478383 TI - Word learning by preschoolers with specific language impairment: effect of phonological or semantic cues. AB - PURPOSE: This study investigated whether phonological or semantic encoding cues promoted better word learning for children with specific language impairment (SLI) and whether this treatment differentially affected children with SLI and normal language (NL). METHOD: Twenty-four preschoolers ages 4;0 (years;months) to 5;11 with SLI and 24 age- and gender-matched children with NL participated. The between-group factor was language group (NL, SLI) and within-group factors were language modality (comprehension, recognition, production) and treatment condition (phonological, semantic). Word learning was assessed during fast mapping, word learning, and post-testing with trials to criterion calculated for the number of words learned. A drawing task assessed the change in semantic representation of words. RESULTS: The SLI group comprehended more words in the semantic condition and produced more words in the phonological condition, but the NL group performed similarly in both. The NL group required significantly fewer trials than the SLI group to comprehend words in the semantic and phonological conditions and to produce words in the semantic condition, but between-group differences for production were not significant for the phonological condition. CONCLUSIONS: The results suggest that preschoolers with SLI may benefit from cues that highlight the phonological or semantic properties of words but that different cues may aid different aspects of word learning. PMID- 16478384 TI - The effect of semantic representation on toddlers' word retrieval. AB - PURPOSE: This study tested the hypothesis that depth of semantic representation influences toddlers' word retrieval. METHOD: Nineteen toddlers participated under 3 word learning conditions in this longitudinal study. Gestures cued attention to object shape (SHP) or function (FNC) in the experimental conditions. No semantic cue was provided under a control condition (CTL). Word learning conditions occurred on each of 3 days. On the 4th day, word retrieval was assessed across 3 levels of scaffolding (uncued picture naming, cued picture naming, picture recognition). Evidence of semantic representation was provided at fast and slow mapping intervals. RESULTS: Less scaffolding was necessary for word retrieval (uncued and cued naming) under experimental conditions than under the CTL condition. However, more SHP than FNC condition targets were retrieved for uncued picture naming. This latter difference may be related to the superior fast mapping of targets under the SHP condition. Toddlers stated object functions (slow mapping) comparably in the experimental conditions, but this was superior to CTL condition performance. CONCLUSIONS: Word retrieval is a continuous behavior that is positively influenced by semantic representation. Semantic knowledge of objects can be enriched by shape or function gestures, thereby improving toddlers' object word productions. Shape cues appear to be more effective for this purpose. PMID- 16478385 TI - Nonword repetition and language development in 4-year-old children with and without a history of early language delay. AB - PURPOSE: This study examined the usefulness of the Nonword Repetition Test (NRT; C. Dollaghan & T. F. Campbell, 1998) with 4-year-old children and the relationship among the NRT, language, and other aspects of mental processing. METHOD: The NRT was administered to 64 children at 4 years of age; 44 had a history of typical language development (HTD), and 20 had a history of language delay (HLD) at 16 months of age. Study 1 compared methods of scoring phoneme errors to determine whether the NRT was appropriate for this age group. Study 2 examined whether the NRT differentiated HTD from HLD. Study 3 examined the relations among scores on the NRT and standardized tests of language and mental processing. RESULTS: The NRT was found to be appropriate for 4-year-old children. Although all children had normal language abilities at the time of the study, the NRT (and several aspects of language and mental processing) differentiated between HTD and HLD. Relations among the NRT and other measures of language and mental processing were different from those previously reported, an unexpected finding that is inconsistent with traditional accounts of working memory and its relation to language development. Potential explanations are explored, and some directions for future research are suggested. PMID- 16478386 TI - Perceiving nonnative vowels: the effect of context on perception as evidenced by event-related brain potentials. AB - In 2 experiments, the authors examined the electrophysiological auditory responses of monolingual French listeners to American English vowel contrasts as a function of the surrounding vowel context. The context was determined on the basis of behavioral results (C. Meunier, C. Frenck-Mestre, T. Lelekov-Boissard, & M. Le Besnaris, 2003, 2004). In the 1st experiment, where the vowel /I/ was placed in a context in which it could easily be discriminated from the surrounding vowels (82% /i/ and 3% /ae/), the electrophysiological response to this vowel showed both acoustic and phonemic responses in line with behavioral results. In the 2nd experiment, where the same vowel /i/ was placed in a difficult context (82% /epsilon/ and 3% /ae/), the electrophysiological response of French participants to this vowel revealed a greatly reduced phonemic response, showing assimilation of the vowel to the surrounding context, again in line with behavioral results. The results of a 3rd control experiment with American participants showed both an acoustic and a phonemic response to the vowel /i/ in the difficult context (82% /epsilon/ and 3% /ae/). This pattern demonstrates the fluctuations in perception as a function of context, and hints at a supple system that may be modified through experience. PMID- 16478387 TI - Phonological awareness in deaf children who use cochlear implants. AB - A short-term longitudinal study was conducted to investigate possible benefits of cochlear implant (CI) use on the development of phonological awareness in deaf children. Nineteen CI users were tested on 2 occasions. Two groups of deaf children using hearing aids were tested once: 11 profoundly deaf and 10 severely deaf children. A battery of tests was designed to investigate syllable, rhyme, and phoneme awareness. Syllable awareness in the CI users was equivalent to that of the severely deaf group, and rhyme and phoneme awareness was similar to that of the profoundly deaf children using hearing aids. CI use affords some benefit to the development of phonological awareness. The results from this study indicate that this enhancement is first observable at the syllable level. PMID- 16478388 TI - Visual attention in deaf and normal hearing adults: effects of stimulus compatibility. AB - Visual perceptual skills of deaf and normal hearing adults were measured using the Eriksen flanker task. Participants were seated in front of a computer screen while a series of target letters flanked by similar or dissimilar letters was flashed in front of them. Participants were instructed to press one button when they saw an H, and another button when they saw an N. Targets H and N were flashed with flanking letters that were either H or N, creating response compatible and response-incompatible arrays. Flankers were presented at different distances from the targets and reaction times were measured. In the present study, reaction times were significantly faster for the hearing group than for the deaf group. However, the hearing group had significantly more errors on this task than the deaf group, suggesting that the deaf participants may have been more deliberate in their responses. In addition, the deaf group revealed a significantly greater interference effect than the hearing group at a parafoveal (i.e., 1.0 degrees ) eccentricity. These findings suggest that deaf individuals may allocate their visual resources over a wider range than those with normal hearing. PMID- 16478392 TI - Short communication: low prevalence of genotypic drug resistance mutations among antiretroviral-naive HIV type 1 patients in Malaysia. AB - To assess the prevalence of mutations associated with drug resistance in antiretroviral-naive patients in Kuala Lumpur, Malaysia, genotypic resistance testing was conducted among drug-naive HIV-1 patients attending the University Malaya Medical Center (UMMC) between July 2003 and June 2004. Reverse transcriptase (RT) and protease genes of plasma virions were sequenced from 100 individuals. The majority of the patients were recently diagnosed. Codons 20-255 of the RT and 1-96 of the protease gene were examined for major and minor mutations associated with antiretroviral resistance reported by the International AIDS Society- USA (IAS-USA) Drug Resistance Mutations Group. The prevalence of patients with at least one major mutation conferring drug resistance was 1%, with only one patient having a Y181C amino acid substitution in the RT gene that confers high-level resistance to nevirapine and delavirdine. Minor mutations were detected in high prevalence in the protease gene. Amino acid substitutions I13V, E35D, and M36I were associated with CRF01_AE while L63P, V77I, and I93L were associated with subtype B. Baseline prevalence of major mutations associated with resistance to antiretroviral drugs was low among antiretroviral-naive HIV-1 patients, suggesting that routine drug resistance testing may be unnecessary for all individuals newly diagnosed with HIV or all patients beginning antiretroviral therapy. PMID- 16478394 TI - Predictive factors of hyperlipidemia in HIV-infected subjects receiving lopinavir/ritonavir. AB - We studied 382 multiexperienced HIV-infected patients followed up for > or =3 months after starting lopinavir/ritonavir (LPV/r) to identify the factors predicting hypertriglyceridemia and high non-HDL cholesterol levels (triglycerides > or =200 mg/dl and/or non-HDL cholesterol > or =190 mg/dl) after 6 and 12 months of LPV/r exposure. The predictors of hypertriglyceridemia were higher baseline triglyceride levels [OR: 2.28 (95% CI: 1.67-3.12) for each additional 100 mg/dl; p = 0.001], the total duration of antiretroviral treatment [OR: 1.26 (95% CI: 1.12-1.41) for each additional year; p = 0.01], CDC stage C (OR: 2.06; 95% CI: 1.24-3.88; p = 0.02), and male gender (OR: 2.52; 95% CI: 1.42 4.74; p = 0.02); intravenous drug abusers seem less likely to develop the event (OR: 0.52; 95% CI: 0.37-0.92; p = 0.03). The predictors of high non-HDL cholesterol levels were higher baseline levels [OR: 3.92 (95% CI: 1.92-6.24) for each additional 100 mg/dl; p = 0.001) and the combination of NRTIs and NNRTIs with LPV/r (OR: 1.83; 95% CI: 1.10-3.69; p = 0.03). The 75 patients stopping LPV/r showed a significant reduction in median triglyceride and non-HDL cholesterol levels after 3 months of 39 mg/dl and 20 mg/dl (p = 0.01 for both), respectively. Patients with high triglyceride and non- HDL cholesterol levels at the start of LPV/r treatment are at higher risk of developing hyperlipidemia. PMID- 16478393 TI - Ethnic variations in the prevalence of metabolic bone disease among HIV-positive patients with lipodystrophy. AB - A high prevalence of metabolic bone disease and osteonecrosis among HIV+ patients on highly active antiretroviral therapy (HAART) has been reported in predominantly white cohorts. We examined bone health in an ethnically diverse cohort of 23 African-Americans and 21 non-African-Americans who were mean (standard deviation) age 45 (7) years old, 66% male, and on HAART for 34 (28) months. Non-African-Americans were more likely to have osteopenia or osteoporosis (59%) compared to African-Americans (26%) (p = 0.09). The prevalence of vitamin D insufficiency (< 34 ng/ml) and elevated i-PTH (>65 pg/ml) was 79% and 20%, respectively. Higher mean urinary N-telopeptide levels were found in non-African Americans [58 (34) nmol BCE/mmol] compared to African-Americans [41 (18) nmol BCE/mmol] (p = 0.09). Magnetic resonance imaging identified one African-American subject (3%) with bilateral asymptomatic hip osteonecrosis. Our findings suggest that the burden of metabolic bone disease in HIV+ patients with HAART-associated lipodystrophy may be greater in whites than in African-Americans. Studies to examine ethnic variations in bone metabolism are necessary to devise optimal interventions. PMID- 16478395 TI - Liver enzyme elevation in hepatitis C virus (HCV)-HIV-coinfected patients prior to and after initiating HAART: role of HCV genotypes. AB - Transaminase elevation is frequently seen in hepatitis C virus (HCV)-HIV coinfected patients receiving antiretroviral therapy (ART), representing an increase in the immune response against HCV and being one of the mechanisms proposed to be involved. There is a report claiming that HCV genotype 3 is an independent risk factor. Our objectives were to assess the incidence of liver toxicity in an HIV-HCV-coinfected population with relatively preserved cellular immunity, and the role of HCV genotypes in the elevation of liver enzymes, both at baseline and after initiating ART. All HIV(+) patients with positive anti-HCV serology and CD4(+) cell counts above 100/mm(3) who began triple ART were identified, and their HCV-RNA levels and HCV genotype were determined. Liver enzymes were determined at baseline and bimonthly during follow-up. Of anti-HCV patients 147 were included, 128 (87.1%) of whom had detectable plasma HCV-RNA. HCV-1 and HCV-4 genotypes were found to confer an increased probability of having at baseline transaminases within normal limits over the other genotypes. Severe transaminase elevations (grades 3 and 4) occurred in 5/124 patients (4.0%), all with high pre-HAART ALT and positive HCV-RNA levels. Multivariate analysis showed that patients with genotype HCV-3 had a 3.27 times higher risk of developing HAART-related transaminase elevations of any grade. In conclusion, subjects with the HCV-1 genotype more often had transaminases within normal limits at baseline. The incidence of severe transaminase elevation after initiating ART was very low (4%) in this HIV(+) population with relatively preserved cellular immunity. HCV genotype 3 was identified as a risk factor for the development of transaminase elevation of any grade. PMID- 16478396 TI - CD8+ cell noncytotoxic anti-HIV response: restoration by HAART in the late stage of infection. AB - Highly active antiretroviral therapy (HAART) is currently the best HIV infection management strategy. However, its effects on the CD8+ T cell noncytotoxic anti HIV response (CNAR) are not well known. We investigated if HAART has different effects on CNAR in patients at the intermediate and late stages of HIV infection. Untreated healthy HIV-infected subjects with a mean CD4+ T cell count of 606 cells/microl were examined as a reference group. Plasma viral load, CD4+ T cell count, and CNAR activity were measured at baseline and regular intervals for at least 48 weeks following initiation of HAART. Baseline CNAR activity in all subjects correlated inversely with viral load and directly with CD4 T+ cell counts. The level of CNAR in the latestage group was significantly lower than in the intermediate-stage and the healthy reference group (p < 0.01). Following initiation of HAART, substantial increases in CD4+ T cell counts and decreases in viral loads were observed in both groups, indicating treatment success. CNAR activity was found to be increased significantly during HAART, but only in the late-stage group (p < 0.01). This increase in CD8+ cell function was seen within 4 weeks of treatment initiation and resulted in levels of CNAR activity almost equal to those observed in the healthy reference subjects. Our findings suggest a beneficial effect on CNAR in those individuals with reduced activity, typically in late-stage infection. PMID- 16478397 TI - Analysis of CCR5, CCR2, CX3CR1, and SDF1 polymorphisms in HIV-positive treated patients: impact on response to HAART and on peripheral T lymphocyte counts. AB - Although polymorphisms of chemokine genes (SDF1, stromal cell-derived factor-1 and RANTES, regulated on activation, normal T cell expressed and secreted) and chemokine-receptor genes (CCR5, CCR2, CX(3)CR1) were shown to be associated with sensitivity to HIV infection and untreated HIV disease progression, their association with the response to highly active antiretroviral therapy (HAART) remains unclear. To explore the possible influence of such polymorphisms on the evolution of AIDS in treated patients, we have studied SDF1-3'A, CCR5Delta32, CCR2-64I, CX(3)CR1-249I, and CX(3)CR1-280M polymorphisms in HIV-infected patients under HAART (n = 169). We studied the evolution of plasma virus load and peripheral T lymphocyte counts in these patients up to 3 years after the initiation of HAART. We observed that some of the genetic polymorphisms studied had an impact on the evolution of these two parameters. After 1 year of HAART, patients with a virological response (undetectable plasma HIV-1 RNA) have a higher frequency of the homozygous SDF1-3'A genotype than other patients (p = 0.005). Similarly, patients with a CD4 increase of over 200/mm(3) from baseline after 1 year of HAART display higher frequencies of homozygous SDF1-3'A (p = 0.035) and homozygous CX(3)CR1-280M genotypes (p = 0.04) than other patients. Moreover, we showed that the CX(3)CR1- 280M allele was associated with higher peripheral CD4+ T cell counts not only in HIV+ patients but also in healthy controls (p = 0.003). PMID- 16478400 TI - Env gp120 sequence analysis of HIV type 1 strains from diverse areas of the brain shows preponderance of CCR5 usage. AB - In this study diverse areas of the autopsied brain of 12 HIV-infected patients with and without dementia were analyzed. All brain samples were obtained at autopsy through prior consent. Env C2-V5 region was PCR amplified and sequenced and compared between different brain regions within the same patient and also between patients to find changes, which can discriminate between patients with and without dementia and also identify motifs responsible for coreceptor-mediated entry of HIV into the CNS. For this, the Env gp120 hypervariable V3 region (35 amino acid residues) was subjected to position scoring matrix analyses (PSSM) for predicting coreceptor usage in the brain. These predictions based on the V3 loop sequence were absolutely consistent with the biologically determined viral phenotype at least for the samples, which were successful for virus culture. These data clearly show that the PSSM correlates can be unambiguously applied in determining viral phenotype for entry. The most notable observation is that of 69 V3 region sequences analyzed from 12 patients from diverse brain regions, 64 showed CCR5 usage (93%) as opposed to only five using CXCR4. Comparison of the V3 loop charge failed to show any correlation between charge and coreceptor usage. Given that cells of macrophage lineage predominate in the CNS and also facilitate HIV entry into the CNS, the preponderance of CCR5 usage in brain-derived HIV strains from patients with and without dementia may have important clinical implications. PMID- 16478399 TI - Full-length genome analysis of human immunodeficiency virus type 1 subtype C in Brazil. AB - The most prevalent HIV-1 clade in the global epidemics is C, and this clade is also becoming important in the Brazilian epidemics. In this study, we characterized HIV-1 subtype C variants by sequencing their near full-length genomes. DNA was extracted from six samples previously classified in our laboratory as subtype C on the basis of partial genome sequencing. Amplification was carried out by overlapping PCR followed by direct sequencing. Phylogenetic analysis of full length genomes confirmed that all isolates belonged to subtype C, which formed a highly supported monophyletic cluster and showed a nucleotide distance of 5.4%. The core promoter of all isolates contained three NF-kappaB binding motifs. Our results suggest that subtype C viruses circulating in Brazil were likely introduced recently from a unique point source. The independent clustering of Brazilian subtype C on the phylogenetic tree suggests the profile of an ideal local candidate for the development of a single subtype vaccine. PMID- 16478398 TI - Thymic function in severely immunodeficient HIV type 1-infected patients receiving stable and effective antiretroviral therapy. AB - The role of the thymus in long-term immune reconstitution has not been addressed in HIV patients who were severely immunodeficient prior to successful treatment with combination antiretroviral therapy (ART). Adult HIV-1 patients (n = 78) with nadir CD4+ T cell counts <100 T cells/microl, at least 12 months on ART and 6 months of complete viral suppression (<50 HIV RNA copies/ml) were selected from a patient database. The cohort was divided according to current CD4+ T cell counts and patients from the lowest (n = 15) and highest (n = 12) tertiles were studied. Thymic volume was assessed by spiral computed tomography. Naive (CD45RA+CD62L+) and replicating (Ki67+) T cells were quantitated by flow cytometry, T cell receptor excision circles (TREC) were assessed by real-time PCR, and serum IL-7 and testosterone by immunoassay. Patients with low CD4+ T cell counts had smaller thymuses [0(0-5.3) vs. 3.5(0-15.6) cm(3), p = 0.04] and were more likely to have no detectable thymus. They had similar proportions of replicating cells, but fewer naive CD4+ and CD8+ T cells and less TREC in CD4+ and CD8+ T cells/ml of blood than patients with high CD4+ T cell counts. However, some patients with no detectable thymus had high numbers of naive and TREC-bearing T cells. Thus, the recovery of CD4+ T cells in severely immunodeficient HIV patients with a virological response to ART is probably limited by thymic function. However, the data are consistent with extrathymic T cell production contributing to the naive T cell pool in some patients. PMID- 16478401 TI - Identification and molecular characterization of subsubtype A4 in central Africa. AB - Phylogenetic analysis on partial env sequences of HIV-1-positive samples obtained from sentinel population groups in HIV serosurveillance studies in 1997 and 2002 in the Democratic Republic of Congo (DRC) revealed a high genetic diversity in the number of cocirculating subtypes/CRFs as well as intrasubtype diversity. A cluster of three strains that formed a distinct and well-separated group within the subtype A radiation was observed. To determine a more precise structure of these viruses, the full-length genomes were sequenced. Phylogenetic tree and bootscan analysis showed that all three newly characterized viruses formed a new nonrecombinant lineage that was more closely related to subtype A. The SUDI (Subtype Distance) program showed that the distances of the newly derived HIV-1 sequences to subtype A references fell in the range of distances previously characterized for subsubtypes. According to current nomenclature rules, we have thus designated this new lineage as subsubtype A4. Analysis of viral sequences from other African countries suggests that the A4 strains seem restricted to DRC where they already circulated at the onset of the HIV/AIDS epidemic. PMID- 16478402 TI - Individual HIV type 1 envelope-specific T cell responses and epitopes do not segregate by virus subtype. AB - HIV-1 vaccines are often designed to target one or several virus subtype(s). They therefore include antigens (e.g., env or env/gag/pol) from each targeted subtype to elicit subtype-directed immunity. To determine if individual T cells respond to HIV-1 antigens in a subtype-directed manner, we selected four T cell hybridomas, each representative of a different immunodominant response toward a subtype B envelope. Hybridomas were tested for responses toward 20 subtype B envelope proteins and one protein each from subtypes A, C, and D. None of the hybridomas cross-reacted with all subtype B envelopes, yet three responded to a non-B protein. Core epitopes and flanking regions affected responsiveness. This lack of subtype-directed activity was corroborated by analyses of the Los Alamos database; like immune responses, epitope distributions were not dictated by subtype. Results highlight the difficulty of predicting immune responses based on subtype alone and encourage considerations of antigenic disparity in addition to subtype disparity during HIV-1 vaccine design. PMID- 16478403 TI - Emergence of a three codon deletion in gag p17 in HIV type 1 subtype C long-term survivors, and general population spread. AB - In a population-based study in northern Malawi we investigated HIV-1 subtype C gag and env gene sequences associated with long-term survival. DNA samples were available from 31 individuals surviving between population surveys carried out in the 1980s and 1990s. Most survivors with paired sequences dating from the 1980s and the 1990s had a three codon deletion in the gag p17 region of the sequence retrieved from the sample collected in the 1990s that was not present in the sequence from the same individual dating from the 1980s. This deletion was also not present in any other 1980s sequences from Malawi, but was common in samples collected in Malawi in the 1990s. The deletion is equivalent to the loss of three amino acids in the D helix region of the gag protein, and may be associated with longer survival and onward transmission. PMID- 16478404 TI - HIV type 1 pol gene diversity and antiretroviral drug resistance mutations in the Democratic Republic of Congo (DRC). AB - To study recombination and the natural polymorphism in pol of HIV-1 strains in the Democratic Republic of Congo (DRC) we sequenced the protease and RT genes for 70 HIV-1 strains previously characterized in the env V3-V5 region from a sentinel surveillance study in 2002. For 41 of the 70 (58.6%) strains, the same subtype/ CRF designations were observed in pol and env. Twenty-three (32.9%) of 70 pol sequences were complex recombinants involving two to five subtypes as well as fragments that could not be classified into any of the known subtypes. All subtypes were involved in recombination events. Unclassified (U) and env subtype H strains were very likely to be recombinant strains. Overall, many minor mutations were identified in the protease sequences. Although at the time of our study ARV use was not yet widespread in DRC, three strains were identified with one major mutation associated with drug resistance: L90M and M46L in protease and K103N in RT. PMID- 16478405 TI - Phylogenetic analysis of Brazilian HIV type 1 subtype D strains: tracing the origin of this subtype in Brazil. AB - HIV-1 Subtype D occurs mainly in East and Central African countries, especially Uganda, where the prevalence of HIV-1 infection is among the highest in the world. We present the phylogenetic analysis of one nonautochthonous and four autochthonous (including a near full-length genome) Brazilian HIV-1 subtype D strains identified in Rio de Janeiro State, where subtypes B, F1, and BF1 recombinants predominate. Phylogenetic inferences using maximum likelihood were applied on a near-full length genome and on concatenated gag, protease, reverse transcriptase, integrase, C2V3/env, gp41, and nef segments. Sequences from an Angolan immigrant showed close genetic similarity with a strain described in Finland, from an HIV patient of African origin, whereas all four autochthonous Brazilian sequences clustered with South African strains, where subtype D occurs only in isolated cases. Our results suggest the successful introduction and circulation in Brazil of closely related HIV-1 subtype D strains, possibly of South African origin. PMID- 16478408 TI - Norwegian Centre for Telemedicine. PMID- 16478409 TI - Differential resource utilization benefits with Internet-based care coordination in elderly veterans with chronic diseases associated with high resource utilization. AB - Our objective was to evaluate in a demonstration project whether our T-Care Program, telecare management via an Internet-based home-messaging device, reduces resource utilization by patients with congestive heart failure (CHF), diabetes mellitus (DM), and chronic obstructive pulmonary disease (COPD). Study participants were drawn from a group of high resource-utilizing veterans over age 60 and included 19 patients with CHF, 23 with DM, and 17 with COPD. Data were gathered on hospital admissions, bed days of care (BDOC), outpatient admissions, and emergency department visits 6 months before and 6 months after enrollment in the telecare program. Nonparametric tests examined pre- and postintervention effects. For patients with CHF, significant decreases were found with T-Care in total emergency department visits (30 to 10, p = 0.03) and hospital admissions (20 to 8, p = 0.03). The decrease in BDOC (179 to 53) was not significant (p = 0.07). Outpatient visits were unchanged (71 to 83, p = 0.38). There were no significant changes for patients with COPD: the apparent BDOC decrease (115 to 46) was not significant (p = 0.24). The outpatient visits by patients with DM decreased significantly (199 to 143, p = 0.03), but no significance was found for changes in their emergency department visits, hospital admissions, and BDOC. The apparent BDOC increase (38 to 198, p = 0.23) was related to two patients with extended stays for an amputation and mitral valve surgery. We found that telecare models may reduce resource utilization in elderly patients with chronic diseases, especially in patients with CHF. Because of the small sample size and lack of controls, larger and more carefully designed follow-up trials are needed to determine cost efficiency for different chronic diseases, and the relative value of the interpersonal contact versus the technological components of this care coordination model. PMID- 16478410 TI - Applying heuristic evaluation to improve the usability of a telemedicine system. AB - The development of a telemedicine system should not only take advantage of technological advances but also pay close attention to users and the human issues involved. In this paper we examine the utility of heuristic evaluation in improving the usability of a digital emergency medical services (EMS) system equipped on an ambulance. The digital EMS system used advanced communication technologies to help remotely located trauma specialists gain access to patient data in real-time and direct life-saving measures in a timely fashion. To improve its usability, three experts inspected prototypes of the system according to 14 software usability heuristics. The analyses revealed information on the prevalence, severity, and nature of heuristic violations in the user interface design. The results were subsequently utilized to guide the iterative software design process. A comparison between two consecutive prototypes showed that the second design had only half as many usability violations as the first prototype and had considerable improvement in a number of usability heuristic categories. The validity of heuristic evaluation was examined in an ethnographic study of paramedics using a prototype of the system in their work environment. Users' task performances partially verified heuristic evaluation results. However, they also revealed problems that were not identified in heuristic evaluation but only became prominent during field observation. In conclusion, we argue that usability should be given high priority in the development of a telemedicine system, and that heuristic evaluation can be an effective and efficient way to identify usability problems in the early stage of software development. PMID- 16478411 TI - A two-period assessment of changes in specialist contact in a high-risk pregnancy telemedical program. AB - The purpose was to examine the organizational impact of a state-wide high-risk pregnancy telemedical system, Antenatal and Neonatal Guidelines, Education and Learning System (ANGELS), after the first year of its roll out. The focus is on several aspects of system organization, including the volume and diversity of patient-based telemedical consultations and weekly telemedical case discussions, telephone consultations, and changes in the pattern of birth-related patient transports. Individual data on patient transports and associated hospital days, provider-specialist telephone calls, and telemedical consultations were collected for two time periods: December 2002-May 2003 (prior to initiation of ANGELS), and December 2003-May 2004 (postinitiation of ANGELS). Different statistical tests were constructed to compare the two periods as appropriate. Significant increases were observed in the volume and geographic diversity of telemedical consultations and the volume of telephone consultations. There was a moderate, but nonsignificant decrease in the number of maternal transports to University of Arkansas School of Medical Sciences (UAMS), and the average length of stay decreased. The type of specialist-provider and specialist-patient contact has changed as the ANGELS high-risk pregnancy telemedical system has evolved over the first year. We conclude that the rollout of the ANGELS program is changing the shape of high-risk patient care in Arkansas, and we attribute that to an evolving collegial network between specialists and generalists. PMID- 16478412 TI - Evaluation of a hands-free wireless communication device in the perioperative environment. AB - The objective of this study was to evaluate the efficiency and reliability of a hands-free voice over Internet protocol (VOIP) communication system in the perioperative environment. Two surveys were administered to anesthesiologists and operating room (OR) nurses working at an academic medical center. Providers were queried by alphanumeric pages or VOIP queries during OR work shifts to measure communication response times. Providers, responding to the query, were asked to verbally complete a system performance survey to capture information regarding their workload and work environment at the time of the query. A user feedback survey was independently administered in writing to a convenience sample of OR providers to obtain information regarding provider communication preferences, concerns, and recommendations. OR providers responded to communication queries four times faster when using VOIP compared to alphanumeric pagers. Providers found VOIP to be much less reliable than conventional pager-telephone systems. Dead spots in the 802.11b network and errors in speaker recognition were frequently cited as sources of system failures. Providers also expressed concern in maintaining confidentiality of patient data or other clinical data communicated using this system. The results of this study suggest that VOIP is still a developing technology but one that is currently viable in the clinical setting. The technology can be used efficiently and securely in health care if users are given the proper training its functions and capabilities. PMID- 16478413 TI - A Web-based telemedicine system for diabetic retinopathy screening using digital fundus photography. AB - The purpose was to design and implement a Web-based telemedicine system for diabetic retinopathy screening using digital fundus cameras and to make the software publicly available through Open Source release. The process of retinal imaging and case reviewing was modeled to optimize workflow and implement use of computer system. The Web-based system was built on Java Servlet and Java Server Pages (JSP) technologies. Apache Tomcat was chosen as the JSP engine, while MySQL was used as the main database and Laboratory of Neuro Imaging (LONI) Image Storage Architecture, from the LONI-UCLA, as the platform for image storage. For security, all data transmissions were carried over encrypted Internet connections such as Secure Socket Layer (SSL) and HyperText Transfer Protocol over SSL (HTTPS). User logins were required and access to patient data was logged for auditing. The system was deployed at Hubert H. Humphrey Comprehensive Health Center and Martin Luther King/Drew Medical Center of Los Angeles County Department of Health Services. Within 4 months, 1500 images of more than 650 patients were taken at Humphrey's Eye Clinic and successfully transferred to King/Drew's Department of Ophthalmology. This study demonstrates an effective architecture for remote diabetic retinopathy screening. PMID- 16478414 TI - Cardiovascular disease prevention for underserved patients using the Internet: bridging the digital divide. AB - For underserved populations, telemedicine can address the high prevalence and suboptimal control of cardiovascular disease (CVD) risk factors. However, Internet access issues may limit the successful application of telemedicine. We tested the hypothesis that computer skills, and not access per se, was the main obstacle to using the Internet for health care. After informed consent, 44 participants with little or no computer experience received 2 hours of training covering 14 basic computer use skills, Internet access, and our telemedicine system. The telemedicine system enables reporting blood pressure, weight, physical activity, cigarette use, provider feedback, personal medication information, and educational information about CVD risk factors. The patient population included 12 males and 32 females. Of this total were 23 African Americans. The average patient age was 60.4 +/- 3 years, and 64% had annual family incomes under 25,000 dollars. Eighty-two percent of the participants averaged 4 or higher (on a scale of 1 to 5) on basic computer skills. Only 11% had an average score below 3. Thirty-seven of 44 participants reported on their health status from a local Internet access site within 10 days. Participants' successful use of the telemedicine system was not correlated with age, gender, education level, or ownership of a computer. Computer skill score had a positive effect on system use. Underserved populations without computer experience or skills and at increased risk for CVD can be educated to use an Internet telemedicine system to communicate health status to their health care providers. Ownership of a computer was not a factor that predicted system use. PMID- 16478415 TI - Computer-based real-time analysis in mobile ocular screening. AB - Mobile ocular telemedicine is potentially an effective method to provide service in medically underserved areas and to screen large populations for abnormalities. Currently, digital images are acquired, stored, and transferred to readers for evaluation, after which the results are provided to the subjects. The transfer of large image files and the timeliness of the subsequent reading of images are significant factors for practical implementation of effective telemedicine screening. This work examines the feasibility of in situ real-time computer analysis of digital images to determine and classify the image results as normal and abnormal. This retrospective study used a photoscreening database of 360 patients ranging in ages from 6 months to 18 years. Computer analysis automatically classified the binocular photorefraction (PR) images, and these PR results were compared to those of the subjective clinical eye examinations provided. With an average processing time of approximately 15 seconds per examinee, the analysis found that the PR results can be categorized as: a positive group that requires referral (186 cases) with a predictive value of 98.9% (2 false-positives); a negative group (144 cases) with a predictive value of 89.6% (15 false-negatives); and an uncertain group (30 cases or 8.3%) that required resolution by readers. The real-time analysis code reduces by approximately 92% the manpower for image grading and electronic transmission at this stage of ocular evaluation. These results indicate the feasibility of this approach. PMID- 16478416 TI - Telementoring in endocrine surgery: preliminary Indian experience. AB - Telemedicine is starting to play an important role in the health field in India. In this case report we describe the successful use of telementoring to remove a parathyroid tumor in a patient with residual hyperparathyroidism after two previous unsuccessful attempts in tumor excision. A 21-yr-old patient crippled with advanced hyperparathyroidism was taken up for third-time exploration at Amrita Institute of Medical Sciences (AIMS), Kochi, with guidance from the Sanjay Gandhi Postgraduate Institute of Medical Sciences (SGPGIMS), Lucknow, using telemedicine technology. These two centers are located 2,500 km apart, and telementoring from the more experienced endocrine surgeons at SGPGIMS resulted in successful tumor localization and removal. For this session both the institutions were provided with a dedicated 512 Kbps very small aperture terminal (VSAT) link and two-way video-audio connectivity. Even though two previous explorations were unsuccessful, with the help of telemedicine technology the same surgeon was successful in locating and removing the tumor. The video and audio quality was of good enough quality for the expert at SGPGIMS to guide the team at AIMS satisfactorily. The patient benefited since he did not have to travel to a far off specialized center for surgery. This case report testifies to the usefulness of telemedicine in the field of surgery, especially in developing countries, which have few medical experts in certain specialized areas. PMID- 16478418 TI - Sees additional factors affecting milk production. PMID- 16478419 TI - A punch list for changing veterinary medicine's public image in the 21st century. PMID- 16478420 TI - What is your diagnosis? Craniodorsal luxation of the atlas from its articulation with the occipital condyles. PMID- 16478421 TI - What is your diagnosis? Abscess developed as a result of scrotal and testicular lesions. PMID- 16478422 TI - ECG of the month. Sinus arrhythmia. PMID- 16478423 TI - Perceived importance and integration of the human-animal bond in private veterinary practice. AB - OBJECTIVE: To determine perceptions of the human-animal bond (HAB) among veterinarians in private practice and evaluate how these veterinarians incorporate the HAB in their practices. DESIGN: Survey. SAMPLE POPULATION: 1,602 veterinarians in private practice in Washington state. PROCEDURE: Participants were contacted and asked to complete a survey. RESULTS: Response rate was 26% (415/1,602). Most respondents agreed that veterinarians will be more successful if they recognize and facilitate the HAB, that facilitating the HAB was important to their practices, that they actively evaluated the degree of bonding between clients and their animals, and that the bonding between a client and his or her animal affected the way they practiced medicine. However, > 50% of respondents did not train veterinary technicians and front office staff members in the HAB or encourage veterinary technicians or front office staff members to learn about the HAB. Fifty-one percent of respondents offered few or no HAB resources to clients. When asked to quantify the importance of 10 nontechnical skills associated with private veterinary practice, respondents ranked communication skills, ethical reasoning, and business management first, second, and third; the HAB was ranked fifth. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that for veterinarians in private practice in Washington state, there is a dichotomy between how important they consider the HAB to be in their practice and the degree to which they facilitate the HAB with regard to communication, training, and client resources. More research on the HAB is necessary to better understand what the HAB encompasses and its implications for private practitioners. PMID- 16478424 TI - Evaluation of veterinary public practice education programs. AB - OBJECTIVE: To describe the number and types of veterinary professional degree and certificate programs providing education in the area of public practice to veterinarians and determine the availability of these programs via distance learning. PROCEDURES: Web-based internet searches were performed for programs for veterinary public practice or public health, population medicine, or Master's degree in Epidemiology. The information reviewed was derived from individual school and program Web sites and from personal e-mail correspondence with school administrators. RESULTS: 17 professional degree and 4 certificate programs were available to provide education and training in the areas of public practice and population medicine to veterinarians. Twelve of these programs have begun since 1998. Of the 17 professional degree programs, 7 are located in the United States and 10 are located in other countries. Nine of the professional degree programs provide education through traditional teaching methods, and 8 provide education and training through distance learning. CONCLUSIONS: During the preceding 5 years, the number of programs available to educate and train veterinarians in the areas of public practice and population medicine has increased. Distance learning is being used to increase capacity and reach a broader audience of veterinarians. With the increase in programs has come an increase in capacity to educate and train veterinarians in the fields of population medicine and public practice. The impact and sustainability of this increased capacity have not been evaluated. PMID- 16478427 TI - Unilateral uveitis in a dog with uveodermatologic syndrome. AB - CASE DESCRIPTION: A 7-year-old Siberian Husky-type dog with heterochromia irides was evaluated because of signs of pain associated with the right eye. CLINICAL FINDINGS: Unilateral panuveitis, iris bombe, and secondary glaucoma were detected in the right eye. Tear production was low bilaterally. Facial and truncal poliosis and vitiligo were also evident; skin biopsy specimens were obtained from the nasal planum. Uveodermatologic syndrome was diagnosed on the basis of histopathologic findings of a lichenoid interface dermatitis and pigmentary incontinence within the dermis. Immunohistochemical analysis was performed on skin samples retrospectively, and findings were inconclusive. TREATMENT AND OUTCOME: Treatment involved topical (ocular) and oral administration of corticosteroids, oral administration of azathioprine, and topical (ocular) administration of a carbonic anhydrase inhibitor and a lacrimostimulant. The secondary glaucoma was refractory to treatment, and the right eye was enucleated. Uveodermatologic syndrome was confirmed via histologic examination of ocular tissues. The left eye remained free of inflammation 16 months after the initial diagnosis. The periocular skin and skin of the nose partially regained pigment, but the hair did not. CLINICAL RELEVANCE: Some breeds in which uveodermatologic syndrome has been reported (eg, Siberian Huskies, Old English Sheepdogs, Australian Shepherds, and Shetland Sheepdogs) often have heterochromia irides. This case highlights the fact that dogs with asymmetric uveal pigmentation may have unilateral ocular changes; therefore, uveodermatologic syndrome should not be excluded as a differential diagnosis on the basis of unilateral clinical signs. PMID- 16478425 TI - Evaluation of bacterial and protozoal contamination of commercially available raw meat diets for dogs. AB - OBJECTIVE: To evaluate bacterial and protozoal contamination of commercially available raw meat diets for dogs. DESIGN: Prospective longitudinal study. SAMPLE POPULATION: 240 samples from 20 raw meat diets for dogs (containing beef, lamb, chicken, or turkey), 24 samples from 2 dry dog foods, and 24 samples from 2 canned dog foods. PROCEDURE: Each product was purchased commercially on 4 dates approximately 2 months apart. Three samples from each product at each sampling period were evaluated via bacterial culture for non-type-specific Escherichia coli (NTSEC), Salmonella enterica, and Campylobacter spp. Antimicrobial susceptibility testing was performed on selected isolates. Polymerase chain reaction assays were used to detect DNA from Cryptosporidium spp, Neospora spp, and Toxoplasma spp in samples obtained in the third and fourth sampling periods. RESULTS: One hundred fifty-three of 288 (53%) samples were contaminated with NTSEC. Both raw and prepared foods contained NTSEC during at least 1 culture period. Salmonella enterica was recovered from 17 (5.9%) samples, all of which were raw meat products. Campylobacter spp was not isolated from any samples. In 91 of 288 (31.6%) samples, there was no gram-negative bacterial growth before enrichment and in 48 of 288 (16.7%) samples, there was no aerobic bacterial growth before enrichment. Susceptibility phenotypes were variable. Cryptosporidium spp DNA was detected in 3 samples. CONCLUSIONS AND CLINICAL RELEVANCE: Bacterial contamination is common in commercially available raw meat diets, suggesting that there is a risk of foodborne illness in dogs fed these diets as well possible risk for humans associated with the dogs or their environments. PMID- 16478428 TI - Evaluation of carbon dioxide laser ablation combined with mitoxantrone and piroxicam treatment in dogs with transitional cell carcinoma. AB - CASE DESCRIPTION: 8 dogs that underwent carbon dioxide (CO2) laser ablation of transitional cell carcinoma in the bladder trigone and proximal portion of the urethra and were also treated with mitotranxone and piroxicam. CLINICAL FINDINGS: Transitional cell carcinoma of the bladder frequently involves the trigone and urethra and can be difficult to manage surgically. Dogs underwent laser ablation of the primary tumor and were treated with mitoxantrone at a dosage of 5 mg/m2)every 3 weeks for 4 treatments. Piroxicam was given at a dosage of 0.3 mg/kg (0.14 mg/lb) once daily for the remaining life of the dog. TREATMENT AND OUTCOME: Median and mean disease-free intervals were 200 and 280 days, respectively. Median and mean survival times were 299 and 411 days, respectively. Adverse treatment effects were observed in 2 dogs; signs included mild, self limiting inappetance and lethargy. The procedure appeared to be well tolerated; all treated dogs had rapid resolution of clinical signs of disease of the lower portion of the urinary tract. CLINICAL RELEVANCE: Although survival times achieved with CO2 laser ablation and treatment with mitoxantrone and piroxicam were similar to survival times associated with chemotherapy alone, resolution of clinical signs was better with the combined treatment. PMID- 16478430 TI - Investigation of antimicrobial use and the impact of antimicrobial use guidelines in a small animal veterinary teaching hospital: 1995-2004. AB - OBJECTIVE: To evaluate patterns of antimicrobial use and the impact of antimicrobial use guidelines at a small animal veterinary teaching hospital. DESIGN: Retrospective study. SAMPLE POPULATION: All antimicrobial prescriptions for dogs and cats admitted to the hospital in 1995 through 2004. PROCEDURE: Pharmacy records were reviewed, and antimicrobial prescriptions for all dogs and cats admitted during the study period were recorded. Amounts of individual drugs dispensed directly to the intensive care and surgical units were determined. Changes in antimicrobial use during the study period were assessed; changes in antimicrobial use patterns in 2000 through 2004 were evaluated to assess the impact of implementation of antimicrobial use guidelines in 2001. RESULTS: There was a significant decrease in prescriptions/1,000 admissions during the study period. From 1995 to 2004, the use of first-generation cephalosporins, fluoroquinolones, penicillins, and trimethoprim-sulfonamides decreased, whereas the use of metronidazole increased. The use of first-generation cephalosporins, fluoroquinolones, and penicillins decreased from 2000 to 2004. First-line drugs accounted for 90.7% of prescriptions during the study period. The use of third line drugs decreased from 2000 to 2004. CONCLUSIONS AND CLINICAL RELEVANCE: Characterization of antimicrobial use is an important step in defining and evaluating the prudent use of antimicrobials. Whereas the true effect of antimicrobial use guidelines is unclear, these results suggest that the guidelines may have had an effect on antimicrobial prescription patterns in this small animal veterinary teaching hospital. Analysis of objective data regarding antimicrobial use and changes in antimicrobial use patterns over time is important in veterinary practices. PMID- 16478432 TI - Survival times for cats with hyperthyroidism treated with iodine 131, methimazole, or both: 167 cases (1996-2003). AB - OBJECTIVE: To compare survival times for cats with hyperthyroidism treated with iodine 131, methimazole, or both and identify factors associated with survival time. DESIGN: Retrospective case series. ANIMALS: 167 cats. PROCEDURE: Medical records of cats in which hyperthyroidism had been confirmed on the basis of high serum thyroxine concentration, results of thyroid scintigraphy, or both were reviewed. RESULTS: 55 (33%) cats were treated with 131I alone, 65 (39%) were treated with methimazole followed by 131I, and 47 (28%) were treated with methimazole alone. Twenty-four of 166 (14%) cats had preexisting renal disease, and 115 (69%) had preexisting hepatic disease. Age was positively correlated (r = 0.4) with survival time, with older cats more likely to live longer. Cats with preexisting renal disease had significantly shorter survival times than did cats without preexisting renal disease. When cats with preexisting renal disease were excluded, median survival time for cats treated with methimazole alone (2.0 years; interquartile range [IQR], 1 to 3.9 years) was significantly shorter than median survival time for cats treated with 131I alone (4.0 years; IQR, 3.0 to 4.8 years) or methimazole followed by 131I (5.3 years; IQR, 2.2 to 6.5 years). CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that age, preexisting renal disease, and treatment type were associated with survival time in cats undergoing medical treatment of hyperthyroidism. PMID- 16478433 TI - Characteristics of cisternal cerebrospinal fluid associated with intracranial meningiomas in dogs: 56 cases (1985-2004). AB - OBJECTIVE: To determine CSF characteristics associated with intracranial meningiomas in dogs. DESIGN: Retrospective case series. ANIMALS: 56 dogs with intracranial meningiomas. PROCEDURES: Medical records of dogs with a histopathologic diagnosis of intracranial meningioma, in which CSF analysis had been performed, were reviewed. Information concerning total nucleated cell counts (TNCCs) and differential nucleated cell counts, RBC counts, and total protein concentration in CSF; seizure history and glucocorticoid administration; and location of meningiomas was recorded. RESULTS: TNCCs < 5 cells/microL were detected in 41 of 56 (73%) dogs; 5 of 56 (9%) dogs had TNCCs > 50 cells/microL. Analysis of CSF revealed predominantly neutrophilic pleocytosis in < 20% of dogs. There was a significant association between meningioma location (caudal portion of the cranial fossa or middle and rostral portion of the cranial fossae) and increased TNCCs (> or = 5 cells/microL). CONCLUSIONS AND CLINICAL RELEVANCE: Results were significantly different from those routinely reported in the veterinary literature. Neutrophilic pleocytosis, especially with TNCCs > 50 cells/microL, was not typical in CSF samples from dogs with intracranial meningiomas. Neutrophilic pleocytosis may not be detected in CSF samples from dogs with meningiomas located within the middle or rostral portion of the cranial fossae. PMID- 16478435 TI - Cytauxzoon felis infection in cats in the mid-Atlantic states: 34 cases (1998 2004). AB - OBJECTIVE: To describe the demographic and clinical characteristics of feline cytauxzoonosis in the mid-Atlantic states and compare the Cytauxzoon felis 18S rRNA gene sequences from affected cats with sequences reported from affected cats in other regions. DESIGN: Retrospective case series. ANIMALS: 34 cats with C. felis infection. PROCEDURE: Medical records of cats in which C. felis infection was diagnosed from May 1998 through June 2004 were reviewed; data collected included signalment, month of diagnosis, geographic location, clinicopathologic abnormalities, medical treatments, outcome, and necropsy findings when applicable. Cytauxzoon felis DNA was amplified, cloned, and sequenced from 4 of these cats and compared with previously reported C. felis DNA sequences. RESULTS: Of 34 C. felis-infected cats, 28 resided in North Carolina, 3 resided in South Carolina, and 3 resided in Virginia; in 32 cats, a diagnosis of C. felis infection was made in April through September. Pancytopenia and icterus were the most common clinicopathologic abnormalities. Thirty-two cats either died or were euthanatized, and 2 cats survived. At 5 veterinary hospitals, multiple cases were identified, and 4 multicat households had > 1 cat infected with C. felis. The 18S rRNA gene sequences characterized in organisms obtained from 4 cats were nearly identical to C. felis DNA sequences reported from other US regions. CONCLUSIONS AND CLINICAL RELEVANCE: Data indicate that veterinarians in the mid-Atlantic region of the United States should consider C. felis infection in cats that become ill with fever, icterus, and pancytopenia or bicytopenia, especially in the spring and summer months. PMID- 16478436 TI - Risk factors associated with renal insufficiency in horses with primary gastrointestinal disease: 26 cases (2000-2003). AB - OBJECTIVE: To identify factors associated with renal insufficiency in colic- or colitis-affected horses with high serum creatinine (SCr) concentrations evaluated at a referral hospital. DESIGN: Retrospective case series. ANIMALS: 167 colic- or colitis-affected horses (88 represented a random sample [hospital population], and 79 had high SCr concentration at initial evaluation [study population]). PROCEDURE: Medical records were reviewed. Data collected included signalment; physical examination, clinicopathologic, and diagnostic findings; and outcome. The study population was categorized on the basis of whether SCr concentration did (AR group; n = 53) or did not (PA group; 26) normalize within 72 hours of fluid therapy. Characteristics of the study and hospital populations were compared. RESULTS: Males and Quarter Horses were significantly overrepresented in the study population. Compared with the hospital population, study-population horses were significantly more likely to have colitis, gastric reflux, and diarrhea at initial evaluation. Initial mean SCr concentration in the PA group was significantly higher than the AR group; identification of gastric reflux, abnormal rectal examination findings, and hypochloremia were significantly associated with persistent azotemia after 72 hours of fluid therapy. Compared with the AR group, PA group horses were 3 times as likely to die or be euthanized. CONCLUSIONS AND CLINICAL RELEVANCE: In colic- or colitis-affected horses, factors associated with renal insufficiency included gastric reflux, abnormal rectal examination findings, or hypochloremia initially; prognosis for horses in which azotemia resolves within 72 hours of treatment appears to be better than for horses with persistent azotemia. PMID- 16478438 TI - Evaluation of diagnostic tests used for detection of bovine viral diarrhea virus and prevalence of subtypes 1a, 1b, and 2a in persistently infected cattle entering a feedlot. AB - OBJECTIVE: To evaluate diagnostic tests used for detection of bovine viral diarrhea virus (BVDV) and determine the prevalence of BVDV subtypes 1a, 1b, and 2a in persistently infected (PI) cattle entering a feedlot. DESIGN: Prospective study. ANIMALS: 21,743 calves. PROCEDURES: Samples were obtained from calves initially testing positive via antigen capture ELISA (ACE) performed on fresh skin (ear notch) specimens, and ACE was repeated. Additionally, immunohistochemistry (IHC) was performed on skin specimens fixed in neutral buffered 10% formalin, and reverse transcriptase PCR (RT-PCR) assay and virus isolation were performed on serum samples. Virus was subtyped via sequencing of the 5' untranslated region of the viral genome. RESULTS: Initial ACE results were positive for BVDV in 88 calves. After subsequent testing, results of ACE, IHC, RT PCR assay, and viral isolation were positive in 86 of 88 calves; results of all subsequent tests were negative in 2 calves. Those 2 calves had false-positive test results. On the basis of IHC results, 86 of 21,743 calves were PI with BVDV, resulting in a prevalence of 0.4%. Distribution of BVDV subtypes was BVDV1b (77.9%), BVDV1a (11.6%), and BVDV2a (10.5%). CONCLUSIONS AND CLINICAL RELEVANCE: Rapid tests such as ACE permit identification and segregation of PI cattle pending results of further tests, thus reducing their contact with the rest of the feedlot population. Although vaccines with BVDV1a and 2a components are given to cattle entering feedlots, these vaccines may not provide adequate protection against BVDV1b. PMID- 16478439 TI - Long-term persistence of multi-drug-resistant Salmonella enterica serovar Newport in two dairy herds. AB - OBJECTIVE: To evaluate the association between maintaining joint hospital and maternity pens and persistence of multi-drug-resistant (MDR) Salmonella enterica serovar Newport on 2 dairy farms. DESIGN: Observational study. SAMPLE POPULATION: Feces and environmental samples from 2 dairy herds. PROCEDURE: Herds were monitored for fecal shedding of S enterica Newport after outbreaks of clinical disease. Fecal and environmental samples were collected approximately monthly from pens housing sick cows and calving cows and from pens containing lactating cows. Cattle shedding the organism were tested serially on subsequent visits to determine carrier status. One farm was resampled after initiation of interventional procedures, including separation of hospital and maternity pens. Isolates were characterized via serotyping, determination of antimicrobial resistance phenotype, detection of the CMY-2 gene, and DNA fingerprinting. RESULTS: The prevalence (32.4% and 33.3% on farms A and B, respectively) of isolating Salmonella from samples from joint hospital-maternity pens was significantly higher than the prevalence in samples from pens housing preparturient cows (0.8%, both farms) and postparturient cows on Farm B (8.8%). Multi-drug-resistant Salmonella Newport was isolated in high numbers from bedding material, feed refusals, lagoon slurry, and milk filters. One cow excreted the organism for 190 days. Interventional procedures yielded significant reductions in the prevalences of isolating the organism from fecal and environmental samples. Most isolates were of the C2 serogroup and were resistant to third generation cephalosporins. CONCLUSIONS AND CLINICAL RELEVANCE: Management practices may be effective at reducing the persistence of MDR Salmonella spp in dairy herds, thus mitigating animal and public health risk. PMID- 16478443 TI - Comparative organization of follicle, accessory cells and spawning anlagen in dynamic semelparous clutch manipulators, the urochordate Oikopleuridae. AB - BACKGROUND INFORMATION: The urochordate appendicularians play a key trophic role in marine ecosystems and are the second largest component of zooplankton after copepods. Part of their success is due to their ability to undergo rapid population blooms in response to changes in primary productivity. Nonetheless, the reproductive biology of this important group remains poorly understood. RESULTS: In the present study, we investigated the organization of male and female germ and accessory somatic cells in the Oikopleuridae. We found that the structure of the ovary had been previously misconstrued as consisting of germ and accessory 'cells' interspersed together, whereas, in fact, the germline exists as a giant transparent syncytium. Somatic follicle cells, integral to regulation of the temporal progression of gametogenesis, could be classified into three types in females and two in males, and we characterized functional gap junctions between follicle cells and the germline syncytium in both sexes. The number of follicle cells per oocyte produced was much reduced in comparison with many commonly studied model organisms. We further identified a novel anlagen that permits spawning of the animal via rupture of the gonad wall, which is obligatory for the release of oocytes, but optional for the release of sperm that usually occurs via the spermiduct. CONCLUSIONS: The organization of the female germline in the Oikopleuridae shares some features of meroistic oogenesis with the arthropod Drosophila, but the process of synchronous oogenesis in these semelparous organisms remains quite distinctive with respect to that previously characterized in the animal kingdom and certainly within the chordate phylum. PMID- 16478441 TI - RanBPM associates with CD39 and modulates ecto-nucleotidase activity. AB - CD39/ecto-NTPDase 1 (nucleoside triphosphate diphosphohydrolase 1) is an ecto nucleotidase that influences P2 receptor activation to regulate vascular and immune cell adhesion and signalling events pivotal in inflammation. Whether CD39 interacts with other membrane or cytoplasmic proteins has not been established to date. Using the yeast two-hybrid system, we note that the N-terminus of CD39 binds to RanBPM (Ran binding protein M; also known as RanBP9), a multi-adaptor scaffolding membrane protein originally characterized as a binding protein for the small GTPase Ran. We confirm formation of complexes between CD39 and RanBPM in transfected mammalian cells by co-immunoprecipitation studies. Endogenous CD39 and RanBPM are also found to be co-expressed and abundant in cell membranes of B lymphocytes. NTPDase activity of recombinant CD39, but not of N-terminus-deleted CD39 mutant, is substantially diminished by RanBPM co-expression in COS-7 cells. The conserved SPRY [repeats in splA and RyR (ryanodine receptor)] moiety of RanBPM is insufficient alone for complete physical and functional interactions with CD39. We conclude that CD39 associations with RanBPM have the potential to regulate NTPDase catalytic activity. This intermolecular interaction may have important implications for the regulation of extracellular nucleotide-mediated signalling. PMID- 16478442 TI - Modulation of Kv2.1 channel gating and TEA sensitivity by distinct domains of SNAP-25. AB - Distinct domains within the SNARE (soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor) proteins, STX1A (syntaxin 1A) and SNAP-25 (synaptosome-associated protein-25 kDa), regulate hormone secretion by their actions on the cell's exocytotic machinery, as well as voltage-gated Ca2+ and K+ channels. We examined the action of distinct domains within SNAP-25 on Kv2.1 (voltage gated K+ 2.1) channel gating. Dialysis of N-terminal SNAP-25 domains, S197 (SNAP-25(1-197)) and S180 (SNAP-25(1-180)), but not S206 (full-length SNAP 25(1-206)) increased the rate of Kv2.1 channel activation and slowed channel inactivation. Remarkably, these N-terminal SNAP-25 domains, acting on the Kv2.1 cytoplasmic N-terminus, potentiated the external TEA (tetraethylammonium) mediated block of Kv2.1. To further examine whether these are effects of the channel pore domain, internal K+ was replaced with Na+ and external K+ was decreased from 4 to 1 mM, which decreased the IC50 of the TEA block from 6.8+/ 0.9 mM to >100 mM. Under these conditions S180 completely restored TEA sensitivity (7.9+/-1.5 mM). SNAP-25 C-terminal domains, SNAP-25(198-206) and SNAP 25(181-197), had no effect on Kv2.1 gating kinetics. We conclude that different domains within SNAP-25 can form distinct complexes with Kv2.1 to execute a fine allosteric regulation of channel gating and the architecture of the outer pore structure in order to modulate cell excitability. PMID- 16478444 TI - Exocellular electron transfer in anaerobic microbial communities. AB - Exocellular electron transfer plays an important role in anaerobic microbial communities that degrade organic matter. Interspecies hydrogen transfer between microorganisms is the driving force for complete biodegradation in methanogenic environments. Many organic compounds are degraded by obligatory syntrophic consortia of proton-reducing acetogenic bacteria and hydrogen-consuming methanogenic archaea. Anaerobic microorganisms that use insoluble electron acceptors for growth, such as iron- and manganese-oxide as well as inert graphite electrodes in microbial fuel cells, also transfer electrons exocellularly. Soluble compounds, like humic substances, quinones, phenazines and riboflavin, can function as exocellular electron mediators enhancing this type of anaerobic respiration. However, direct electron transfer by cell-cell contact is important as well. This review addresses the mechanisms of exocellular electron transfer in anaerobic microbial communities. There are fundamental differences but also similarities between electron transfer to another microorganism or to an insoluble electron acceptor. The physical separation of the electron donor and electron acceptor metabolism allows energy conservation in compounds as methane and hydrogen or as electricity. Furthermore, this separation is essential in the donation or acceptance of electrons in some environmental technological processes, e.g. soil remediation, wastewater purification and corrosion. PMID- 16478445 TI - Structural diversity of bacterial communities in a heavy metal mineralized granite outcrop. AB - This laboratory study of a variably mineralized and hydrothermally altered granite outcrop investigated the influences of rock-surface chemistry and heavy metal content on resident bacterial populations. Results indicated that elevated heavy metal concentrations had a profound impact on bacterial community structure, with strong relationships found between certain ribotypes and particular chemical/heavy metal elements. Automated ribosomal intergenic sequence analysis (ARISA) was used to assess the nature and extent of bacterial diversity, and this was combined with chemical analysis and multivariate statistics to identify the main geochemical factors influencing bacterial community structure. A randomization test revealed significant changes in bacterial structure between samples, while canonical correspondence analysis (CCA) related each individual ARISA profile to linear combinations of the chemical variables (mineralogy, major element and heavy metal concentrations) revealing the geochemical factors that correlated with changes in the ARISA data. anova was performed to further explore interactions between individual ribotypes and chemical/heavy metal composition, and revealed that a high proportion of ribotypes correlated significantly with heavy metals. PMID- 16478446 TI - Diversity and ubiquity of thermophilic methanogenic archaea in temperate anoxic soils. AB - Temperate rice field soil from Vercelli (Italy) contains moderately thermophilic methanogens of the yet uncultivated rice cluster I (RC-I), which become prevalent upon incubation at temperatures of 45-50 degrees C. We studied whether such thermophilic methanogens were ubiquitously present in anoxic soils. Incubation of different rice field soils (from Italy, China and the Philippines) and flooded riparian soils (from the Netherlands) at 45 degrees C resulted in vigorous CH(4) production after a lag phase of about 10 days. The archaeal community structure in the soils was analysed by terminal restriction fragment length polymorphism (T RFLP) targeting the SSU rRNA genes retrieved from the soil, and by cloning and sequencing. Clones of RC-I methanogens mostly exhibited T-RF of 393 bp, but also terminal restriction fragment (T-RF) of 158 and 258 bp length, indicating a larger diversity than previously assumed. No RC-I methanogens were initially found in flooded riparian soils. However, these archaea became abundant upon incubation of the soil at 45 degrees C. Thermophilic RC-I methanogens were also found in the rice field soils from Pavia, Pila and Gapan. However, the archaeal communities in these soils also contained other methanogenic archaea at high temperature. Rice field soil from Buggalon, on the other hand, only contained thermophilic Methanomicrobiales rather than RC-I methanogens, and rice field soil from Jurong mostly Methanomicrobiales and only a few RC-I methanogens. The archaeal community of rice field soil from Zhenjiang almost exclusively consisted of Methanosarcinaceae when incubated at high temperature. Our results show that moderately thermophilic methanogens are common in temperate soils. However, RC-I methanogens are not always dominating or ubiquitous. PMID- 16478447 TI - Selective enrichment and molecular characterization of a previously uncultured Nitrospira-like bacterium from activated sludge. AB - Previously uncultured nitrite-oxidizing bacteria affiliated to the genus Nitrospira have for the first time been successfully enriched from activated sludge from a municipal wastewater treatment plant. During the enrichment procedure, the abundance of the Nitrospira-like bacteria increased to approximately 86% of the total bacterial population. This high degree of purification was achieved by a novel enrichment protocol, which exploits physiological features of Nitrospira-like bacteria and includes the selective repression of coexisting Nitrobacter cells and heterotrophic contaminants by application of ampicillin in a final concentration of 50 microg ml(-1). The enrichment process was monitored by electron microscopy, fluorescence in situ hybridization (FISH) with rRNA-targeted probes and fatty acid profiling. Phylogenetic analysis of 16S rRNA gene sequences revealed that the enriched bacteria represent a novel Nitrospira species closely related to uncultured Nitrospira-like bacteria previously found in wastewater treatment plants and nitrifying bioreactors. The enriched strain is provisionally classified as 'Candidatus Nitrospira defluvii'. PMID- 16478449 TI - Design of Shewanella-specific 16S rRNA primers and application to analysis of Shewanella in a minerotrophic wetland. AB - In this study, an existing probe was used as a polymerase chain reaction (PCR) primer to study iron-reducing members of the genus Shewanella in a minerotrophic wetland where iron reduction had previously been implicated. The probe was found to be non-specific and a new set of PCR primers were developed that were specific for Shewanella. These primers were used to analyse the wetland iron-reducing communities by characterizing 16S rRNA genes amplified from DNA extracted from peat. Polymerase chain reaction clone libraries were screened using restriction fragment length polymorphism and diagnostic operational taxonomic units for Shewanella species were identified. A statistical method was used to determine the coverage of the clone libraries, which was found to be between 83% and 97%. The dominant species in the wetland samples at two geochemically distinct zones were phylogenetically related to the iron-reducing microorganism Shewanella oneidensis. PMID- 16478448 TI - The 'pH optimum anomaly' of intracellular enzymes of Ferroplasma acidiphilum. AB - A wide range of microorganisms, the so-called acidophiles, inhabit acidic environments and grow optimally at pH values between 0 and 3. The intracellular pH of these organisms is, however, close to neutrality or slightly acidic. It is to be expected that enzymatic activities dedicated to extracellular functions would be adapted to the prevailing low pH of the environment (0-3), whereas intracellular enzymes would be optimally active at the near-neutral pH of the cytoplasm (4.6-7.0). The genes of several intracellular or cell-bound enzymes, a carboxylesterase and three alpha-glucosidases, from Ferroplasma acidiphilum, a cell wall-lacking acidophilic archaeon with a growth optimum at pH 1.7, were cloned and expressed in Escherichia coli, and their products purified and characterized. The Ferroplasmaalpha-glucosidases exhibited no sequence similarity to known glycosyl hydrolases. All enzymes functioned and were stable in vitro in the pH range 1.7-4.0, and had pH optima much lower than the mean intracellular pH of 5.6. This 'pH optimum anomaly' suggests the existence of yet-undetected cellular compartmentalization providing cytoplasmic pH patchiness and low pH environments for the enzymes we have analysed. PMID- 16478451 TI - Measurement of Prochlorococcus ecotypes using real-time polymerase chain reaction reveals different abundances of genotypes with similar light physiologies. AB - Prochlorococcus is a marine cyanobacterium which is found at high abundances in world's tropical and subtropical oligotrophic oceans. The genus Prochlorococcus can be divided into two major groups based on light physiology. Both of these groups can be further subdivided into genetically distinct lineages, or ecotypes. Real-time polymerase chain reaction (PCR) assays based on sequence differences in the 16S-23S rDNA internal transcribed spacer or the 23S rDNA were developed to examine the distribution of each ecotype in the field. The real-time PCR assays enabled linear quantification of concentrations ranging from 10 to 4 x 10(5) cells ml(-1). These assays were applied to a stratified water column in the Sargasso Sea. The majority of Prochlorococcus cells above 110 m belonged to the one of the low chlorophyll b/a ratio (high-light adapted) ecotypes, while two types of high chlorophyll b/a ratio (low-light adapted) cells dominated below 110 m. The other three types were found at significantly lower numbers or not detected at all. Differences in the abundance of ecotypes within the major light physiology groupings suggest that other factors, such as nutrient utilization and differential mortality, are driving their relative distributions. Real-time PCR assays will enable further exploration of these factors and temporal and geographic variability in ecotype abundance. PMID- 16478450 TI - Identifying pioneer bacterial species responsible for biofouling membrane bioreactors. AB - More effective control of membrane biofouling in membrane bioreactors (MBRs) lies in the fundamental understanding of the pioneer microorganisms responsible for surface colonization that leads to biofilm formation. In this study, the composition of the planktonic and sessile microbial communities inhabiting four laboratory-scale MBR systems were compared using amplified ribosomal DNA restriction analysis (ARDRA) and 16S ribosomal DNA gene sequencing. The ARDRA results suggest that the microbial communities on membrane surfaces could be very different from the ones in the suspended biomass. Phylogenetic analysis based on the 16S rRNA gene sequences provided a list of bacteria that might be the pioneers of surface colonization on microfiltration membranes. The results further suggested that research on the mechanisms of cell attachment in such an engineering environment could be critical for future development of appropriate biofouling control strategies. PMID- 16478452 TI - Pseudomonas, the dominant polycyclic aromatic hydrocarbon-degrading bacteria isolated from Antarctic soils and the role of large plasmids in horizontal gene transfer. AB - Twenty-two polycyclic aromatic hydrocarbon (PAH)-degrading bacterial strains were isolated from Antarctic soils with naphthalene or phenanthrene as a sole carbon source, while no degrader was obtained from an unpolluted sampling site. Phylogenetic analysis showed that all belonged to the genus Pseudomonas except one that was identified as the genus of Rahnella. Some of them were closely related to previously reported cold-tolerant species, while some were separated in deeply rooted branches and represent new strains. All these strains showed a high efficiency to degrade naphthalene at 4 degrees C, and some additionally degraded phenanthrene. Using degenerate primers and polymerase chain reaction (PCR) amplification, ndo gene encoding naphthalene dioxygenase (NDO) was detected from all the isolates. Phylogenetic analysis grouped these genes into two clusters which shared 94% similarity to each other, and showed about 97% similarity within a cluster. However, no obvious difference was observed with mesophilic ndo genes; this indicates that the host cell is pivotal in cold adaptation. In addition, the mismatch between 16S rRNA and NDO phylogenetic trees strongly indicates horizontal gene transfer among these isolates and may have happened in situ. Further, Southern hybridization and plasmid curing confirmed that ndo genes were located on a large self-transmissible plasmid, which can be transferred to a mesophilic strains. The transconjugants acquired the ability to utilize naphthalene and phenanthrene. Results of this article imply that Pseudomonas plays an important role in PAH biodegradation in Antarctic soils, and the related genes might be originally transferred from outside Antarctica and spread among indigenous species. PMID- 16478453 TI - Analysis of the proteome of Pseudomonas putida KT2440 grown on different sources of carbon and energy. AB - Using 2D electrophoresis the protein expression pattern during growth on carbon sources with different impact on carbon catabolite repression of phenol degradation was analysed in a derivative of Pseudomonas putida KT2440. The cytosolic protein pattern of cells growing on phenol or the non-repressive substrate pyruvate was almost identical, but showed significant differences to that of cells growing with the repressive substrates succinate or glucose. Proteins, which were mainly expressed in the presence of phenol or pyruvate, could be assigned to the functional groups of transport, detoxification, stress response, amino acid, energy, carbohydrate and nucleotide metabolism. The addition of succinate to cells growing with phenol ('shift-up') resulted in the inhibition of the synthesis of these proteins. Proteins with enhanced expression at growth with succinate or glucose were proteins for de novo synthesis of nucleotides, amino acids and enzymes of the TCA cycle. The synthesis of proteins, necessary for phenol catabolism was regulated in different manners following the addition of succinate. Whereas the synthesis of Phl-proteins (subunits of the phenolhydroxylase) only decreased slowly, was the translation of the Cat-proteins (catechol 1,2-dioxygenase, cis,cis-muconate cycloisomerase and muconolactone isomerase) repressed immediately and the synthesis of the Pca-proteins (beta ketoadipate enolactone hydrolase, beta-ketoadipate succinyl-CoA transferase and beta-ketoadipyl CoA thiolase) remained unaffected. PMID- 16478454 TI - Ecophysiology of a group of uncultured Gammaproteobacterial glycogen-accumulating organisms in full-scale enhanced biological phosphorus removal wastewater treatment plants. AB - The presence of glycogen-accumulating organisms (GAOs) in enhanced biological phosphorus removal (EBPR) plants can seriously deteriorate the biological P removal by out-competing the polyphosphate-accumulating organisms (PAOs). In this study, uncultured putative GAOs (the GB group, belonging to the Gammaproteobacteria) were investigated in detail in 12 full-scale EBPR plants. Fluorescence in situ hybridization (FISH) revealed that the biovolume of the GB bacteria constituted 2-6% of total bacterial biovolume. At least six different subgroups of the GB bacteria were found, and the number of dominant subgroups present in each plant varied between one and five. Ecophysiological investigations using microautoradiography in combination with FISH showed that, under aerobic or anaerobic conditions, all subgroups of the GB bacteria could take up acetate, pyruvate, propionate and some amino acids, while some subgroups in addition could take up formate and thymidine. Glucose, ethanol, butyrate and several other organic substrates were not taken up. Glycolysis was essential for the anaerobic uptake of organic substrates. Polyhydroxyalkanoates (PHA) but not polyphosphate (polyP) granules were detected in all GB bacterial cells. Polyhydroxyalkanoate formation after anaerobic uptake of acetate was confirmed by measuring the increase in fluorescence intensity of PHA granules inside GB bacterial cells after Nile blue staining. One GB subgroup was possibly able to denitrify, and several others were able to reduce nitrate to nitrite. PAOs were also enumerated by FISH in the same treatment plants. Rhodocyclus-related PAOs and Actinobacteria-related PAOs constituted up to 7% and 29% of total bacterial biovolume respectively. Rhodocyclus-related PAOs always coexisted with the GB bacteria and showed many physiological similarities. Factors of importance for the competition between the three groups of important bacteria in EBPR plants are discussed. PMID- 16478455 TI - Quantitative, longitudinal profiling of the primate fecal microbiota reveals idiosyncratic, dynamic communities. AB - We used slot blot hybridization, quantitative polymerase chain reaction (qPCR), and flow cytometry microarrays to quantify specific 16S rDNAs in weekly fecal specimens from four monkeys housed in a research vivarium for periods ranging from five to 8 months. Even in these uniformly housed and fed animals the gut microbiota is idiosyncratic, very dynamic on short timescales, and shows significant positive and negative correlations among some bacteria as well as responses to heavy metal exposure. The relative quantification (fmol targets per total fmol bacterial 16S rDNA) afforded by flow cytometry microarrays agreed well with the absolute quantification (nanogram of target DNA per nanogram of fecal DNA) afforded by slot blots and qPCR. We also noted strengths and weaknesses in inter-method comparisons for DNA-based quantification of these complex bacterial communities. PMID- 16478456 TI - Population structure, persistence, and seasonality of autochthonous Escherichia coli in temperate, coastal forest soil from a Great Lakes watershed. AB - The common occurrence of Escherichia coli in temperate soils has previously been reported, however, there are few studies to date to characterize its source, distribution, persistent capability and genetic diversity. In this study, undisturbed, forest soils within six randomly selected 0.5 m2 exclosure plots (covered by netting of 2.3 mm2 mesh size) were monitored from March to October 2003 for E. coli in order to describe its numerical and population characteristics. Culturable E. coli occurred in 88% of the samples collected, with overall mean counts of 16 MPN g(-1), ranging from < 1 to 1657 (n = 66). Escherichia coli counts did not correlate with substrate moisture content, air, or soil temperatures, suggesting that seasonality were not a strong factor in population density control. Mean E. coli counts in soil samples (n = 60) were significantly higher inside than immediately outside the exclosures; E. coli distribution within the exclosures was patchy. Repetitive extragenic palindromic polymerase chain reaction (Rep-PCR) demonstrated genetic heterogeneity of E. coli within and among exclosure sites, and the soil strains were genetically distinct from animal (E. coli) strains tested (i.e. gulls, terns, deer and most geese). These results suggest that E. coli can occur and persist for extended periods in undisturbed temperate forest soils independent of recent allochthonous input and season, and that the soil E. coli populations formed a cohesive phylogenetic group in comparison to the set of fecal strains with which they were compared. Thus, in assessing E. coli sources within a stream, it is important to differentiate background soil loadings from inputs derived from animal and human fecal contamination. PMID- 16478457 TI - The desert of Tataouine: an extreme environment that hosts a wide diversity of microorganisms and radiotolerant bacteria. AB - The phylogenetic diversity of prokaryotic communities exposed to arid conditions in the hot desert of Tataouine (south Tunisia) was estimated with a combination of a culture and - molecular-based analysis. Thirty-one isolates, representative of each dominant morphotypes, were affiliated to Actinobacteria, Firmicutes, Proteobacteria and the CFB group while none related to Archaea. Analysis of 16S rRNA gene libraries revealed the presence of species related to Bacteria and Archaea. Sequences related to Archaea were all affiliated to the non-thermophilic Crenarchaeota subgroup. Bacterial sequences were dominated by Proteobacteria, Actinobacteria and Acidobacteria; a few sequences were distributed among eight others phyla, including Thermus/Deinococcus relatives. A correlation between tolerance to desiccation and to radiation has been demonstrated for the radiotolerant bacteria Deinococcus radiodurans. Because bacteria living in the hot desert of Tataouine are one way or another tolerant to desiccation, we investigate whether they could also be tolerant to radiation. Exposition of soil samples to intense gamma radiation yields Bacillus, Thermus/Deinococcus and alpha Proteobacteria relatives. Four of these strains correspond to radiotolerant species as revealed by evaluation of the resistance levels of the individual cultures. A detailed analysis of the resistance levels for two Thermus/Deinococcus and two alpha-Proteobacteria relatives revealed that they correspond to new radiotolerant species. PMID- 16478458 TI - Molecular analysis of the rstR and orfU genes of the CTX prophages integrated in the small chromosomes of environmental Vibrio cholerae non-O1, non-O139 strains. AB - The ctxAB genes encoding cholera toxin, reside in the genome of a filamentous bacteriophage CTXphi. The presence of CTX prophage in non-epidemic environmental Vibrio cholerae strains is rare. The CTX prophage, the lysogenic form of CTXphi in V. cholerae, is comprised of the 'RS2' and the 'Core'. Analysis of the rstR gene present in the RS2 region of the CTX prophage revealed the presence of new alleles of the prophages in four environmental non-O1, non-O139 strains VCE22 (O36), VCE228 (O27), VCE232 (O4) and VCE233 (O27), and the CTX prophages are located in the small chromosomes. Phylogenetic analysis based on the nucleotide sequences of the rstR and orfU (present in the core) genes of these prophages placed them in a single unique cluster, which is distally located compared with that of epidemic V. cholerae O1 strains. Further analysis indicated that the genome of the prophage present in the strain VCE22 is devoid of the ctxAB genes, called pre-CTX prophage and the strain also possess the toxin-coregulated pilus protein coding gene tcpA of classical type, another important pathogenicity determining locus of the epidemic V. cholerae strains. Comparative analysis of the nucleotide sequences of the rstR and orfU genes indicated that the pre-CTX prophage of VCE22 might be the progenitor of new alleles of the CTX prophages present in these environmental strains. PMID- 16478459 TI - Microbial degradation of street dust polycyclic aromatic hydrocarbons in microcosms simulating diffuse pollution of urban soil. AB - Diffuse pollution with polycyclic aromatic hydrocarbons (PAHs) of topsoil in urban regions has caused increasing concerns in recent years. We simulated diffuse pollution of soil in microcosms by spiking sandy topsoil (A-horizon) and coarse, mineral subsoil (C-horizon) with street dust (PM63) isolated from municipal street sweepings from central Copenhagen. The microbial communities adapted to PAH degradation in microcosms spiked with street dust in both A horizon and C-horizon soils, in spite of low PAH-concentrations. The increased potential for PAH degradation was demonstrated on several levels: by slowly diminishing PAH-concentrations, increased mineralization of 14C-PAHs, increasing numbers of PAH degraders and increased prevalence of nah and pdo1 PAH degradation genes, i.e. the microbial communities quickly adapted to PAH degradation. Three- and 4-ring PAHs from the street dust were biodegraded to some extent (10-20%), but 5- and 6-ring PAHs were not biodegraded in spite of frequent soil mixing and high PAH degradation potentials. In addition to biodegradation, leaching of 2-, 3 and 4-ring PAHs from the A-horizon to the C-horizon seems to reduce PAH-levels in surface soil. Over time, levels of 2-, 3- and 4-ring PAHs in surface soil may reach equilibrium between input and the combination of biodegradation and leaching. However, levels of the environmentally critical 5- and 6-ring PAHs will probably continue to rise. We presume that sorption to black carbon particles is responsible for the persistence and low bioaccessibility of 5- and 6-ring PAHs in diffusely polluted soil. PMID- 16478460 TI - Surveying biotransformations with a la carte genetic traps: translating dehydrochlorination of lindane (gamma-hexachlorocyclohexane) into lacZ-based phenotypes. AB - The ability of the product of a desired reaction to activate a bacterial transcriptional regulator was exploited to develop genetic traps that render the catalytic activity born by a DNA clone into a selectable/scorable phenotype. We established this strategy with a system to expose the activity of dehydrochlorinases acting upon gamma-hexachlorocyclohexane (gamma-HCH or lindane). To this end, the effector-binding protein, XylR, was evolved by gene shuffling plus mutagenic polymerase chain reaction to be optimally responsive to the major product of gamma-HCH dehydrochlorination, 1,2,4-trichlorobenzene (TCB). We then derived Escherichia coli strains that constitutively expressed the modified XylR variant (named XylR5) and had lacZ under control of the Pu promoter, which is activated by XylR. A robotic beta-galactosidase assay indicated that when the resulting strain was transformed with a linA+ clone (expressing a gamma-HCH dehydrochlorinase from Sphingomonas paucimobilis UT26), it had levels of beta-galactosidase that were dependent on the gamma-HCH concentration. This a la carte host thus translated the conversion of gamma-HCH to TCB into upregulation of lacZ. An alternate host additionally expressing LacY grew efficiently on lactose only when LacZ was upregulated in a fashion dependent on TCB or other effectors of XylR5. These results demonstrated the power of deriving a host for the genetic scrutiny, rather than enzymatic screening, of clones expressing a given catabolic enzyme. PMID- 16478461 TI - Analysis of the transmission of Salmonella spp. through generations of pet snakes. AB - Besides the 'classical' animals known as reservoirs for Salmonella spp., like poultry and cattle, reptiles have emerged as a significant source of human salmonellae infections during the last years. Reptile-associated salmonellae frequently cause severe clinical courses including fatalities due to septicaemia and meningitis. Therefore, it is of major priority to develop measures which may help preventing cases of reptile-associated salmonellae. However, as a first step the epidemiology of salmonellae in reptiles must be understood. Therefore, in this study a population analysis of the salmonellae of two female snakes was performed and the pattern of inheritance of salmonellae to their offspring was investigated. It is demonstrated that adult snakes usually harbour a population of concurrent salmonellae serovars. Colonization of their offspring during pregnancy and birth is a significant way of transmission causing 65% of the newborn to be positive for salmonellae. The effectiveness of the transmission does not seem to be due only to the frequency of a certain serovar, because the most prevailing strain of one female snake was not detectable in any of her offspring. PMID- 16478462 TI - Utilization of capsaicin and vanillylamine as growth substrates by Capsicum (hot pepper)-associated bacteria. AB - Capsaicin contributes to the organoleptic attributes of hot peppers. Here, we show that capsaicin is utilized as a growth nutrient by certain bacteria. Enrichment cultures utilizing capsaicin were successfully initiated using Capsicum-derived plant material or leaves of tomato (a related Solanaceae) as inocula. No other sources of inoculum examined yielded positive enrichments. Of 25 isolates obtained from enrichments: all utilized 8-methylnonanoic acid; nine were found capable of degrading capsaicin as sole carbon and energy source; 11 were found capable of utilizing vanillylamine; but only two strains could use either of these latter two compounds as sole nitrogen source. Phylogenetic analysis of capsaicin degraders revealed them to be strains of Variovorax and Ralstonia, whereas the vanillylamine degraders were strains of Pseudomonas and Variovorax. Neither of the two strains isolated from one enrichment culture originally inoculated with dried pepper fruit was capable of using capsaicin as sole carbon and nitrogen source. However, good growth was achieved under such conditions when the two isolates, a strain of Variovorax paradoxusThat degraded capsaicin when provided with ammonium, and a vanillylamine degrading strain of Pseudomonas putida, were cultured together. A cross-feeding of capsaicin-derived carbon and nitrogen between members of pepper-associated consortia is proposed. PMID- 16478464 TI - Theodor Bucher Lecture. Metabolomics, modelling and machine learning in systems biology - towards an understanding of the languages of cells. Delivered on 3 July 2005 at the 30th FEBS Congress and the 9th IUBMB conference in Budapest. AB - The newly emerging field of systems biology involves a judicious interplay between high-throughput 'wet' experimentation, computational modelling and technology development, coupled to the world of ideas and theory. This interplay involves iterative cycles, such that systems biology is not at all confined to hypothesis-dependent studies, with intelligent, principled, hypothesis-generating studies being of high importance and consequently very far from aimless fishing expeditions. I seek to illustrate each of these facets. Novel technology development in metabolomics can increase substantially the dynamic range and number of metabolites that one can detect, and these can be exploited as disease markers and in the consequent and principled generation of hypotheses that are consistent with the data and achieve this in a value-free manner. Much of classical biochemistry and signalling pathway analysis has concentrated on the analyses of changes in the concentrations of intermediates, with 'local' equations - such as that of Michaelis and Menten v=(Vmax x S)/(S+K m) - that describe individual steps being based solely on the instantaneous values of these concentrations. Recent work using single cells (that are not subject to the intellectually unsupportable averaging of the variable displayed by heterogeneous cells possessing nonlinear kinetics) has led to the recognition that some protein signalling pathways may encode their signals not (just) as concentrations (AM or amplitude-modulated in a radio analogy) but via changes in the dynamics of those concentrations (the signals are FM or frequency-modulated). This contributes in principle to a straightforward solution of the crosstalk problem, leads to a profound reassessment of how to understand the downstream effects of dynamic changes in the concentrations of elements in these pathways, and stresses the role of signal processing (and not merely the intermediates) in biological signalling. It is this signal processing that lies at the heart of understanding the languages of cells. The resolution of many of the modern and postgenomic problems of biochemistry requires the development of a myriad of new technologies (and maybe a new culture), and thus regular input from the physical sciences, engineering, mathematics and computer science. One solution, that we are adopting in the Manchester Interdisciplinary Biocentre (http://www.mib.ac.uk/) and the Manchester Centre for Integrative Systems Biology (http://www.mcisb.org/), is thus to colocate individuals with the necessary combinations of skills. Novel disciplines that require such an integrative approach continue to emerge. These include fields such as chemical genomics, synthetic biology, distributed computational environments for biological data and modelling, single cell diagnostics/bionanotechnology, and computational linguistics/text mining. PMID- 16478465 TI - Effects of sequestration on signal transduction cascades. AB - The building blocks of most signal transduction pathways are pairs of enzymes, such as kinases and phosphatases, that control the activity of protein targets by covalent modification. It has previously been shown [Goldbeter A & Koshland DE (1981) Proc Natl Acad Sci USA 78, 6840-6844] that these systems can be highly sensitive to changes in stimuli if their catalysing enzymes are saturated with their target protein substrates. This mechanism, termed zero-order ultrasensitivity, may set thresholds that filter out subthreshold stimuli. Experimental data on protein abundance suggest that the enzymes and their target proteins are present in comparable concentrations. Under these conditions a large fraction of the target protein may be sequestrated by the enzymes. This causes a reduction in ultrasensitivity so that the proposed mechanism is unlikely to account for ultrasensitivity under the conditions present in most in vivo signalling cascades. Furthermore, we show that sequestration changes the dynamics of a covalent modification cycle and may account for signal termination and a sign-sensitive delay. Finally, we analyse the effect of sequestration on the dynamics of a complex signal transduction cascade: the mitogen-activated protein kinase (MAPK) cascade with negative feedback. We show that sequestration limits ultrasensitivity in this cascade and may thereby abolish the potential for oscillations induced by negative feedback. PMID- 16478466 TI - Cloning and characterization of CBL-CIPK signalling components from a legume (Pisum sativum). AB - The studies on calcium sensor calcineurin B-like protein (CBL) and CBL interacting protein kinases (CIPK) are limited to Arabidopsis and rice and their functional role is only beginning to emerge. Here, we present cloning and characterization of a protein kinase (PsCIPK) from a legume, pea, with novel properties. The PsCIPK gene is intronless and encodes a protein that showed partial homology to the members of CIPK family. The recombinant PsCIPK protein was autophosphorylated at Thr residue(s). Immunoprecipitation and yeast two hybrid analysis showed direct interaction of PsCIPK with PsCBL, whose cDNA and genomic DNA were also cloned in this study. PsCBL showed homology to AtCBL3 and contained calcium-binding activity. We demonstrate for the first time that PsCBL is phosphorylated at its Thr residue(s) by PsCIPK. Immunofluorescence/confocal microscopy showed that PsCBL is exclusively localized in the cytosol, whereas PsCIPK is localized in the cytosol and the outer membrane. The exposure of plants to NaCl, cold and wounding co-ordinately upregulated the expression of PsCBL and PsCIPK genes. The transcript levels of both genes were also coordinately stimulated in response to calcium and salicylic acid. However, drought and abscisic acid had no effect on the expression of these genes. These studies show the ubiquitous presence of CBL/CIPK in higher plants and enhance our understanding of their role in abiotic and biotic stress signalling. PMID- 16478467 TI - Structure and influence on stability and activity of the N-terminal propeptide part of lung surfactant protein C. AB - Mature lung surfactant protein C (SP-C) corresponds to residues 24-58 of the 21 kDa proSP-C. A late processing intermediate, SP-Ci, corresponding to residues 12 58 of proSP-C, lacks the surface activity of SP-C, and the SP-Ci alpha-helical structure does not unfold in contrast to the metastable nature of the SP-C helix. The NMR structure of an analogue of SP-Ci, SP-Ci(1-31), with two palmitoylCys replaced by Phe and four Val replaced by Leu, in dodecylphosphocholine micelles and in ethanol shows that its alpha-helix vs. that of SP-C is extended N terminally. The Arg-Phe part in SP-Ci that is cleaved to generate SP-C is localized in a turn structure, which is followed by a short segment in extended conformation. Circular dichroism spectroscopy of SP-Ci(1-31) in microsomal or surfactant lipids shows a mixture of helical and extended conformation at pH 6, and a shift to more unordered structure at pH 5. Replacement of the N-terminal hexapeptide segment SPPDYS (known to constitute a signal in intracellular targeting) of SP-Ci with AAAAAA results in a peptide that is mainly unstructured, independent of pH, in microsomal and surfactant lipids. Addition of a synthetic dodecapeptide, corresponding to the propeptide part of SP-Ci, to mature SP-C results in slower aggregation kinetics and altered amyloid fibril formation, and reduces the surface activity of phospholipid-bound SP-C. These data suggest that the propeptide part of SP-Ci prevents unfolding by locking the N-terminal part of the helix, and that acidic pH results in structural disordering of the region that is proteolytically cleaved to generate SP-C. PMID- 16478468 TI - Cytochrome P450 Cyp4x1 is a major P450 protein in mouse brain. AB - A novel cytochrome P450, CYP4x1, was identified in EST databases on the basis of similarity to a conserved region in the C-helix of the CYP4A family. The human and mouse CYP4x1 cDNAs were cloned and found to encode putative cytochrome P450 proteins. Molecular modelling of CYP4x1 predicted an unusual substrate binding channel for the CYP4 family. Expression of human CYP4x1 was detected in brain by EST analysis, and in aorta by northern blotting. The mouse cDNA was used to demonstrate that the Cyp4x RNA was expressed principally in brain, and at much lower levels in liver; hepatic levels of the Cyp4x1 RNA were not affected by treatment with the inducing agents phenobarbital, dioxin, dexamethasone or ciprofibrate, nor were the levels affected in PPARalpha-/- mice. A specific antibody for Cyp4x1 was developed, and shown to detect Cyp4x1 in brain; quantitation of the Cyp4x1 protein in brain demonstrated approximately 10 ng of Cyp4x1 protein.mg(-1) microsomal protein, showing that Cyp4x1 is a major brain P450. Immunohistochemical localization of the Cyp4x1 protein in brain showed specific staining of neurons, choroids epithelial cells and vascular endothelial cells. These data suggest an important role for Cyp4x1 in the brain. PMID- 16478469 TI - Identification of beta-amyrin and sophoradiol 24-hydroxylase by expressed sequence tag mining and functional expression assay. AB - Triterpenes exhibit a wide range of structural diversity produced by a sequence of biosynthetic reactions. Cyclization of oxidosqualene is the initial origin of structural diversity of skeletons in their biosynthesis, and subsequent regio- and stereospecific hydroxylation of the triterpene skeleton produces further structural diversity. The enzymes responsible for this hydroxylation were thought to be cytochrome P450-dependent monooxygenase, although their cloning has not been reported. To mine these hydroxylases from cytochrome P450 genes, five genes (CYP71D8, CYP82A2, CYP82A3, CYP82A4 and CYP93E1) reported to be elicitor inducible genes in Glycine max expressed sequence tags (EST), were amplified by PCR, and screened for their ability to hydroxylate triterpenes (beta-amyrin or sophoradiol) by heterologous expression in the yeast Saccharomyces cerevisiae. Among them, CYP93E1 transformant showed hydroxylating activity on both substrates. The products were identified as olean-12-ene-3beta,24-diol and soyasapogenol B, respectively, by GC-MS. Co-expression of CYP93E1 and beta-amyrin synthase in S. cerevisiae yielded olean-12-ene-3beta,24-diol. This is the first identification of triterpene hydroxylase cDNA from any plant species. Successful identification of a beta-amyrin and sophoradiol 24-hydroxylase from the inducible family of cytochrome P450 genes suggests that other triterpene hydroxylases belong to this family. In addition, substrate specificity with the obtained P450 hydroxylase indicates the two possible biosynthetic routes from triterpene-monool to triterpene-triol. PMID- 16478470 TI - Activation of the Torpedo nicotinic acetylcholine receptor. The contribution of residues alphaArg55 and gammaGlu93. AB - The Torpedo nicotinic acetylcholine receptor is a heteropentamer (alpha2betagammadelta) in which structurally homologous subunits assemble to form a central ion pore. Viewed from the synaptic cleft, the likely arrangement of these subunits is alpha-gamma-alpha-delta-beta lying in an anticlockwise orientation. High affinity binding sites for agonists and competitive antagonists have been localized to the alpha-gamma and alpha-delta subunit interfaces. We investigated the involvement of amino acids lying at an adjacent interface (gamma alpha) in receptor properties. Recombinant Torpedo receptors, expressed in Xenopus oocytes, were used to investigate the consequences of mutating alphaArg55 and gammaGlu93, residues that are conserved in most species of the peripheral nicotinic receptors. Based on homology modeling, these residues are predicted to lie in close proximity to one another and it has been suggested that they may form a salt bridge in the receptor's three-dimensional structure (Sine et al. 2002 J Biol Chem277, 29 210-29 223). Although substitution of alphaR55 by phenylalanine or tryptophan resulted in approximately a six-fold increase in the EC50 value for acetylcholine activation, the charge reversal mutation (alphaR55E) had no significant effect. In contrast, the replacement of gammaE93 by an arginine conferred an eight-fold increase in the potency for acetylcholine induced receptor activation. In the receptor carrying the double mutations, alphaR55E-gammaE93R or alphaR55F-gammaE93R, the potency for acetylcholine activation was partially restored to that of the wild-type. The results suggest that, although individually these residues influence receptor activation, direct interactions between them are unlikely to play a major role in the stabilization of different conformational states of the receptor. PMID- 16478471 TI - Peptides corresponding to helices 5 and 6 of Bax can independently form large lipid pores. AB - Proteins of the B-cell lymphoma protein 2 (Bcl2) family are key regulators of the apoptotic cascade, controlling the release of apoptotic factors from the mitochondrial intermembrane space. A helical hairpin found in the core of water soluble folds of these proteins has been reported to be the pore-forming domain. Here we show that peptides including any of the two alpha-helix fragments of the hairpin of Bcl2 associated protein X (Bax) can independently induce release of large labelled dextrans from synthetic lipid vesicles. The permeability promoted by these peptides is influenced by intrinsic monolayer curvature and accompanied by fast transbilayer redistribution of lipids, supporting a toroidal pore mechanism as in the case of the full-length protein. However, compared with the pores made by complete Bax, the pores made by the Bax peptides are smaller and do not need the concerted action of tBid. These data indicate that the sequences of both fragments of the hairpin contain the principal physicochemical requirements for pore formation, showing a parallel between the permeabilization mechanism of a complex regulated protein system, such as Bax, and the much simpler pore forming antibiotic peptides. PMID- 16478472 TI - The enzyme-binding region of human GM2-activator protein. AB - The GM2-activator protein (GM2AP) is an essential cofactor for the lysosomal degradation of ganglioside GM2 by beta-hexosaminidase A (HexA). It mediates the interaction between the water-soluble exohydrolase and its membrane-embedded glycolipid substrate at the lipid-water interface. Functional deficiencies in this protein result in a fatal neurological storage disorder, the AB variant of GM2 gangliosidosis. In order to elucidate this cofactor's mode of action and identify the surface region of GM2AP responsible for binding to HexA, we designed several variant forms of this protein and evaluated the consequences of these mutations for lipid- and enzyme-binding properties using a variety of biophysical and functional studies. The point mutants D113K, M117V and E123K showed a drastically decreased capacity to stimulate HexA-catalysed GM2 degradation. However, surface plasmon resonance (SPR) spectroscopy showed that the binding of these variants to immobilized lipid bilayers and their ability to solubilize lipids from anionic vesicles were the same as for the wild-type protein. In addition, a fluorescence resonance energy transfer (FRET)-based assay system showed that these variants had the same capacity as wild-type GM2AP for intervesicular lipid transfer from donor to acceptor liposomes. The concentration dependent effect of these variants on hydrolysis of the synthetic substrate 4 methylumbelliferyl-2-acetamido-2-deoxy-6-sulfo-beta-D-glucopyranoside (MUGS) indicated a weakened association with the enzyme's alpha subunit. This identifies the protein region affected by these mutations, the single short alpha helix of GM2AP, as the major determinant for the interaction with the enzyme. These results further confirm that the function of GM2AP is not restricted to a biological detergent that simply disrupts the membrane structure or lifts the substrate out of the lipid plane. In contrast, our data argue in favour of the critical importance of distinct activator-hexosaminidase interactions for GM2 degradation, and corroborate the view that the activator/lipid complex represents the true substrate for the degrading enzyme. PMID- 16478473 TI - Insulin-dependent phosphorylation of DPP IV in liver. Evidence for a role of compartmentalized c-Src. AB - Dipeptidyl peptidase IV (DPP IV, CD26, EC 3.4.14.5) serves as a model aimed at elucidating protein sorting signals. We identify here, by MS, several tyrosine phosphorylated proteins in a rat liver Golgi/endosome (G/E) fraction including DPP IV. We show that a pool of DPP IV is tyrosine-phosphorylated. Maximal phosphorylation was observed after 2 min following intravenous insulin injection. DPP IV coimmunoprecipitated with the cellular tyrosine kinase Src (c-Src) with maximal association also observed after 2 min following insulin injection. DPP IV was found phosphorylated after incubation of nonsolubilized G/E membranes with [gamma-32P]ATP. The c-Src inhibitor PP2 inhibited DPP IV phosphorylation. Oriented proteolysis experiments indicate that a large pool of c-Src is protected in G/E fractions. Following injection of the protein-tyrosine phosphatase inhibitor bpV(phen), DPP IV levels markedly decreased by 40% both in plasma membrane and G/E fractions. In the fraction designated Lh, DPP IV levels decreased by 50% 15 min following insulin injection. Therefore, a pool of DPP IV is tyrosine-phosphorylated in an insulin-dependent manner. The results suggest the presence of a yet to be characterized signalling mechanism whereby DPP IV has access to c-Src-containing signalling platforms. PMID- 16478474 TI - The essential tyrosine-containing loop conformation and the role of the C terminal multi-helix region in eukaryotic phenylalanine ammonia-lyases. AB - Besides the post-translationally cyclizing catalytic Ala-Ser-Gly triad, Tyr110 and its equivalents are of the most conserved residues in the active site of phenylalanine ammonia-lyase (PAL, EC 4.3.1.5), histidine ammonia-lyase (HAL, EC 4.3.1.3) and other related enzymes. The Tyr110Phe mutation results in the most pronounced inactivation of PAL indicating the importance of this residue. The recently published X-ray structures of PAL revealed that the Tyr110-loop was either missing (for Rhodospridium toruloides) or far from the active site (for Petroselinum crispum). In bacterial HAL ( approximately 500 amino acids) and plant and fungal PALs ( approximately 710 amino acids), a core PAL/HAL domain ( approximately 480 amino acids) with >or= 30% sequence identity along the different species is common. In plant and fungal PAL a approximately 100-residue long C-terminal multi-helix domain is present. The ancestor bacterial HAL is thermostable and, in all of its known X-ray structures, a Tyr83-loop-in arrangement has been found. Based on the HAL structures, a Tyr110-loop-in conformation of the P. crispum PAL structure was constructed by partial homology modeling, and the static and dynamic behavior of the loop-in/loop-out structures were compared. To study the role of the C-terminal multi-helix domain, Tyr-loop in/loop-out model structures of two bacterial PALs (Streptomyces maritimus, 523 amino acids and Photorhabdus luminescens, 532 amino acids) lacking this C terminal domain were also built. Molecular dynamics studies indicated that the Tyr-loop-in conformation was more rigid without the C-terminal multi-helix domain. On this basis it is hypothesized that a role of this C-terminal extension is to decrease the lifetime of eukaryotic PAL by destabilization, which might be important for the rapid responses in the regulation of phenylpropanoid biosynthesis. PMID- 16478475 TI - Structural and functional roles for beta-strand 7 in the alpha-crystallin domain of p26, a polydisperse small heat shock protein from Artemia franciscana. AB - Oviparous development in the extremophile crustacean, Artemia franciscana, generates encysted embryos which enter a profound state of dormancy, termed diapause. Encystment is marked by the synthesis of p26, a polydisperse small heat shock protein thought to protect embryos from stress. In order to elucidate structural/functional relationships within p26 and other polydisperse small heat shock proteins, and to better define the protein's role during diapause, amino acid substitutions R110G, F112R, R114A and Y116D were generated within the p26 alpha-crystallin domain by site-directed mutagenesis. These residues were chosen because they are highly conserved across species boundaries, and molecular modelling indicates that they are part of a key structural interface between dimers. The F112R mutation, which had the greatest impact on oligomerization, placed two charged residues at the p26 dimer-dimer interface, demonstrating the importance of beta-strand 7 in tetramer formation. All mutated versions of p26 were less able than wild-type p26 to confer thermotolerance on transformed bacteria and they exhibited diminished chaperone action in three in vitro assays; however, all variants retained protective activity. This apparent stability of p26 may, by prolonging effective chaperone life in vivo, enhance embryo stress resistance. All substitutions modified p26 intrinsic fluorescence, surface hydrophobicity and secondary structure, and the pronounced changes in variant R114A, as indicated by these physical measurements, correlated with the greatest loss of function. Although mutation R114A had the greatest effect on p26 chaperoning, it had the least on oligomerization. These results demonstrate that in contrast to many other small heat shock proteins, p26 effectiveness as a chaperone is independent of oligomerization. The results also reinforce the idea, occasioned by modelling, that R114 is removed slightly from dimer-dimer interfaces. Moreover, beta-strand 7 is shown to have an important role in oligomerization of p26, a function first proposed for this structural element upon crystallization of wheat Hsp16.9, a small heat shock protein with different quaternary structure. PMID- 16478477 TI - Stage specific expression of poly(malic acid)-affiliated genes in the life cycle of Physarum polycephalum. Spherulin 3b and polymalatase. AB - Polymalic acid is receiving interest as a unique biopolymer of the plasmodia of mycetozoa and recently as a biogenic matrix for the synthesis of devices for drug delivery. The acellular slime mold Physarum polycephalum is characterized by two distinctive growth phases: uninucleated amoebae and multinucleated plasmodia. In adverse conditions, plasmodia reversibly transform into spherules. Only plasmodia synthesize poly(malic acid) (PMLA) and PMLA-hydrolase (polymalatase). We have performed suppression subtractive hybridization (SSH) of cDNA from amoebae and plasmodia to identify plasmodium-specific genes involved in PMLA metabolism. We found cDNA encoding a plasmodium-specific, spherulin 3a-like polypeptide, NKA48 (spherulin 3b), but no evidence for a PMLA-synthetase encoding transcript. Inhibitory RNA (RNAi)-induced knockdown of NKA48-cDNA generated a severe reduction in the level of PMLA suggesting that spherulin 3b functioned in regulating the level of PMLA. Unexpectedly, cDNA of polymalatase was not SSH selected, suggesting its presence also in amoebae. Quantitative PCR then revealed low levels of mRNA in amoebae, high levels in plasmodia, and also low levels in spherules, in agreement with the expression under transcriptional regulation in these cells. PMID- 16478476 TI - The catalysis of the SARS 3C-like protease is under extensive regulation by its extra domain. AB - The 3C-like protease of the severe acute respiratory syndrome (SARS) coronavirus has a C-terminal extra domain in addition to the chymotrypsin-fold adopted by picornavirus 3C proteases hosting the complete catalytic machinery. Previously we identified the extra domain to be involved in enzyme dimerization which has been considered essential for the catalytic activity. In an initial attempt to map out the extra-domain residues critical for dimerization, we have systematically generated 15 point mutations, five deletions and one triple mutation and subsequently characterized them by enzymatic assay, dynamic light scattering, CD and NMR spectroscopy. The results led to identification of four regions critical for enzyme dimerization. Interestingly, Asn214Ala mutant with a significant tendency to form a monomer still retained approximately 30% activity, indicating that the relationship between the activity and dimerization might be very complex. Very surprisingly, two regions (one over Ser284-Thr285-Ile286 and another around Phe291) were discovered on which Ala-mutations significantly increased the enzymatic activities. Based on this, a super-active triple-mutant STI/A with a 3.7-fold activity enhancement was thus engineered by mutating residues Ser284, Thr285 and Ile286 to Ala. The dynamic light scattering, CD and NMR characterizations indicate that the wild-type (WT) and STI/A mutant share similar structural and dimerization properties, thus implying that in addition to dimerization, the extra domain might have other mechanisms to regulate the catalytic machinery. We rationalized these results based on the enzyme structure and consequently observed an interesting picture: the majority of the dimerization-critical residues plus Ser284-Thr285-Ile286 and Phe291 are clustered together to form a nano-scale channel passing through the central region of the enzyme. We therefore speculate that this channel might play a role in relaying regulatory effects from the extra domain to the catalytic machinery. PMID- 16478478 TI - Identification and characterization of two dipeptidyl-peptidase III isoforms in Drosophila melanogaster. AB - Dipeptidyl-peptidase III (DPP III) hydrolyses small peptides with a broad substrate specificity. It is thought to be involved in a major degradation pathway of the insect neuropeptide proctolin. We report the purification and characterization of a soluble DPP III from 40 g Drosophila melanogaster. Western blot analysis with anti-(DPP III) serum revealed the purification of two proteins of molecular mass 89 and 82 kDa. MS/MS analysis of these proteins resulted in the sequencing of 45 and 41 peptide fragments, respectively, confirming approximately 60% of both annotated D. melanogaster DPP III isoforms (CG7415-PC and CG7415-PB) predicted at 89 and 82 kDa. Sequencing also revealed the specific catalytic domain HELLGH in both isoforms, indicating that they are both effective in degrading small peptides. In addition, with a probe specific for D. melanogaster DPP III, northern blot analysis of fruit fly total RNA showed two transcripts at approximately 2.6 and 2.3 kb, consistent with the translation of 89-kDa and 82 kDa DPP III proteins. Moreover, the purified enzyme hydrolyzed the insect neuropeptide proctolin (Km approximately 4 microm) at the second N-terminal peptide bound, and was inhibited by the specific DPP III inhibitor tynorphin. Finally, anti-(DPP III) immunoreactivity was observed in the central nervous system of D. melanogaster larva, supporting a functional role for DPP III in proctolin degradation. This study shows that DPP III is in actuality synthesized in D. melanogaster as 89-kDa and 82-kDa isoforms, representing two native proteins translated from two alternative mRNA transcripts. PMID- 16478479 TI - The crystal structure of a plant 2C-methyl-D-erythritol 4-phosphate cytidylyltransferase exhibits a distinct quaternary structure compared to bacterial homologues and a possible role in feedback regulation for cytidine monophosphate. AB - The homodimeric 2C-methyl-D-erythritol 4-phosphate cytidylyltransferase contributes to the nonmevalonate pathway of isoprenoid biosynthesis. The crystal structure of the catalytic domain of the recombinant enzyme derived from the plant Arabidopsis thaliana has been solved by molecular replacement and refined to 2.0 A resolution. The structure contains cytidine monophosphate bound in the active site, a ligand that has been acquired from the bacterial expression system, and this observation suggests a mechanism for feedback regulation of enzyme activity. Comparisons with bacterial enzyme structures, in particular the enzyme from Escherichia coli, indicate that whilst individual subunits overlay well, the arrangement of subunits in each functional dimer is different. That distinct quaternary structures are available, in conjunction with the observation that the protein structure contains localized areas of disorder, suggests that conformational flexibility may contribute to the function of this enzyme. PMID- 16478480 TI - Inhibition of human ether a go-go potassium channels by Ca2+/calmodulin binding to the cytosolic N- and C-termini. AB - Human ether a go-go potassium channels (hEAG1) open in response to membrane depolarization and they are inhibited by Ca2+/calmodulin (CaM), presumably binding to the C-terminal domain of the channel subunits. Deletion of the cytosolic N-terminal domain resulted in complete abolition of Ca2+/CaM sensitivity suggesting the existence of further CaM binding sites. A peptide array-based screen of the entire cytosolic protein of hEAG1 identified three putative CaM-binding domains, two in the C-terminus (BD-C1: 674-683, BD-C2: 711 721) and one in the N-terminus (BD-N: 151-165). Binding of GST-fusion proteins to Ca2+/CaM was assayed with fluorescence correlation spectroscopy, surface plasmon resonance spectroscopy and precipitation assays. In the presence of Ca2+, BD-N and BD-C2 provided dissociation constants in the nanomolar range, BD-C1 bound with lower affinity. Mutations in the binding domains reduced inhibition of the functional channels by Ca2+/CaM. Employment of CaM-EF-hand mutants showed that CaM binding to the N- and C-terminus are primarily dependent on EF-hand motifs 3 and 4. Hence, closure of EAG channels presumably requires the binding of multiple CaM molecules in a manner more complex than previously assumed. PMID- 16478481 TI - BSE risk assessments in the UK: a risk tradeoff? AB - Risk assessments for bovine spongiform encephalopathy (BSE) should be based on the group risk and not the median individual risk. The group risk is calculated from the arithmetic mean risk, which in the case of dorsal root ganglia, is a factor of 50-fold higher than the median. For environmental routes, the arithmetic mean exposure is sufficient for risk assessment, while for food-borne routes failure to accommodate the variation in exposures to individuals across the UK population could overestimate the group risk considerably. Ignoring prion destruction by cooking could overestimate the food-borne risks still further. The recent estimate for the arithmetic mean cow-to-man species barrier of 4000 does not take into accounts either of these factors and thus may be too high. Until evidence for a threshold dose is demonstrated, public health scientists should avoid assessing safety on the basis of a 'minimum infective dose'. The incubation period observed in cattle-feeding studies, when completed, would continue to increase with decreasing dose below the ID50if there is a threshold or co operative effect. The question is raised of whether fears over BSE in drinking water contributed to the spread of foot-and-mouth disease across the UK in 2001; a risk tradeoff. PMID- 16478482 TI - Biosynthetic requirements for the repair of sublethal membrane damage in Escherichia coli cells after pulsed electric fields. AB - AIMS: The aim was to evaluate the biosynthetic requirements for the repair of sublethal membrane damages in Escherichia coli cells after exposure to pulsed electric fields (PEF). METHODS AND RESULTS: The partial loss of the barrier and homeostatic functions of the cytoplasmic membrane was examined by adding sodium chloride to the recovery media. More than 4 log10 cycles of survivors were sublethally injured after PEF. Repair of such sublethal membrane damages occurred when survivors to PEF were incubated in peptone water for 2 h. Two different types of sublethally injured cells were detected. Whereas a small proportion (<5%) repaired after PEF in less than 2 min, the repair of the remaining 95% injured cells lasted 2 h and was dependent on biosynthetic requirements. The addition of inhibitors such as chloramphenicol, cerulenin, penicillin G, rifampicin and sodium azide to the liquid repair medium showed that the repair required energy and lipid synthesis, and was not dependent on protein, peptidoglican or RNA synthesis. CONCLUSIONS: Cell survival after PEF is dependent on the repair of the cytoplasmic membrane. Requirement of lipid synthesis for the repair of sublethally injured cells confirms that the cytoplasmic membrane is a target directly involved in the mechanism of inactivation by PEF. SIGNIFICANCE AND IMPACT OF THE STUDY: Knowledge about the damages inflicted by PEF might help in the design of more efficient treatments. PMID- 16478483 TI - Genetic diversity of Acacia tortilis ssp. raddiana rhizobia in Tunisia assessed by 16S and 16S-23S rDNA genes analysis. AB - AIMS: In order to understand the genetic diversity of Acacia tortilis ssp. raddiana-rhizobia in Tunisia, isolates from nine geographical locations were obtained and analysed. METHODS AND RESULTS: Characterization using restriction fragment length polymorphism analysis (RFLP) of PCR-amplified 16S rRNA gene and the intergenic spacer (IGS) between the 16S and 23S rRNA genes was undertaken. Symbiotic efficiency of the strains was also estimated. Analysis of the 16S rRNA by PCR-RFLP showed that the isolates were phylogenetically related to Ensifer ssp., Rhizobium tropicii-IIA, and Rhizobium tumefaciens species. Analysis of 16S 23S spacer by PCR-RFLP showed a high diversity of these rhizobia and revealed eleven additional groups, which indicates that these strains are genetically very diverse. Full 16S rRNA gene-sequencing showed that the majority of strains form a new subdivion inside the genera Ensifer, with Ensifer meliloti being its nearest neighbour. Nodulation test performed on the plant host demonstrated differences in the infectivity among the strains. CONCLUSION: Rhizobial populations that nodulate specifically and efficiently Acacia tortilis ssp. raddiana in representative soils of Tunisia is dominated by E. meliloti-like genomospecies. SIGNIFICANCE AND IMPACT OF THE STUDY: This paper provides the first clear characterization and symbiotic efficiency data of rhizobia strains nodulating A. tortilis in Tunisia. PMID- 16478484 TI - Sequence, organization, transcription and regulation of lactose and galactose operons in Lactobacillus rhamnosus TCELL-1. AB - AIMS: Understanding the metabolism of lactose and galactose and their regulation in Lactobacillus rhamnosus. METHODS AND RESULTS: A gene cluster containing nine open reading frames (ORFs) involved in the metabolism of lactose and galactose in Lact. rhamnosus TCELL-1 was sequenced and characterized. The order of the ORFs was lacTEGF and galKETRM. Northern blotting experiments revealed that the gene cluster could be transcribed as one lacTEGF-galKETRM mRNA though three major transcripts (lacTEGF, galKETRM and galETRM) were detected for the gene cluster. The transcription of the lac or gal operon was independently induced in the presence of lactose or galactose. Northern blotting and primer extension experiments found the presence of four putative promoters upstream from the ORFs lacT (lacTp), galK (galKp1 and galKp2) and galE (galEp). The measurements of enzymatic activities of GalK, GalE and GalT suggested that the expression of the gal operon was subjected to a galactose activation and glucose repression mechanism. CONCLUSIONS: In Lact. rhamnosus TCELL-1, the galactose moiety of lactose could be metabolized by two alternative pathways (the Leloir and the tagatose 6-phosphate pathways) whereas galactose metabolism could be mediated by the Leloir pathway. SIGNIFICANCE AND IMPACT OF THE STUDY: This work provides important information about sugar metabolism in Lact. rhamnosus. PMID- 16478485 TI - Molecular characterization of the bacteria adherent to human colorectal mucosa. AB - AIMS: To study large intestinal mucosal bacterial communities by Denaturing Gradient Gel Electrophoresis (DGGE) profiling and sequencing of 16S rRNA gene polymerase chain reaction (PCR) products amplified from DNA extracted from colorectal biopsies taken from healthy individuals. The specific aims were to determine how similar the mucosa-associated bacterial communities are within and between individuals and also to characterize the phylogenetic origin of isolated DGGE bands. METHODS AND RESULTS: Human colorectal biopsies were taken at routine colonoscopy from 33 patients with normal looking mucosa. The DNA was extracted directly from single biopsies and the bacterial 16S rDNA PCR amplified. The PCR products were profiled using DGGE to generate a fingerprint of the dominant members of the bacterial community associated with the biopsy. The reproducibility of this method was high (>98%). Washed and unwashed biopsies gave similar DGGE banding patterns (Median Similarity Coefficient - MSC 96%, InterQuartile Range - IQR 3.0%, n = 5). Adjacent biopsies sampled from the same patient using different forceps gave similar DGGE profiles (MSC 94%, n = 2). Two colorectal biopsies sampled at locations 2-5 cm apart, from each of 18 patients, resulted in very similar profiles (MSC 100%, IQR 2.8%). Biopsies sampled from different locations within the large intestine of the same patient also gave similar DGGE profiles (MSC 98% IQR 3.3%n = 6). Although all patients (n = 33) gave different DGGE profiles, some similarity (c. 34%) was observed between profiles obtained from 15 patients arbitrarily selected. 35 DGGE bands were excised and sequenced. Many were found to be most closely related to uncultured bacterial sequence entries in the Genbank database. Others belonged to typical gut bacterial genera including Bacteroides, Ruminococcus, Faecalibacterium and Clostridium. CONCLUSIONS: Bacterial communities adherent to colorectal mucosa within a normal patient show little variation; in contrast, mucosal bacterial communities sampled from different patients with normal colorectal mucosa show a high degree of variation. SIGNIFICANCE AND IMPACT OF THE STUDY: This research demonstrates that DGGE profiling of 16S rRNA gene PCR products amplified from DNA extracted directly from mucosal samples offers fresh insight into the bacterial communities that are adherent to colorectal mucosa. These findings are important with respect to further studies on the gastrointestinal tract in health and disease. PMID- 16478486 TI - Interference in adhesion of bacteria and yeasts isolated from explanted voice prostheses to silicone rubber by rhamnolipid biosurfactants. AB - AIMS: The effects and extent of adhesion of four different bacterial and two yeast strains isolated from explanted voice prostheses to silicone rubber with and without an adsorbed rhamnolipid biosurfactant layer obtained from Pseudomonasaeruginosa DS10-129 was studied. METHODS AND RESULTS: The ability of rhamnolipid biosurfactant to inhibit adhesion of micro-organisms to silicone rubber was investigated in a parallel-plate flow chamber. The anti-adhesive activity of the biosurfactant at different concentrations was significant against all the strains and depended on the micro-organism tested. The results showed an effective reduction in the initial deposition rates, and the number of bacterial cells adhering after 4 h, for all micro-organisms tested at the 4 g l(-1) undiluted rhamnolipid solution. Maximum initial reduction of adhesion rate (an average of 66%) occurred for Streptococcus salivarius GB 24/9 and Candida tropicalis GB 9/9. The number of cells adhering after 4 h on silicone rubber conditioned with biosurfactant was reduced to 48% for Staphylococcus epidermidis GB 9/6, Strep. salivarius GB 24/9, Staphylococcus aureus GB 2/1 and C. tropicalis GB 9/9 in comparison to controls. Perfusing the flow chamber with biosurfactant containing solution followed by the passage of a liquid-air interface, to investigate detachment of micro-organisms adhering to silicone rubber, produced high detachment (96%) of adhered cells for all micro-organisms studied, except for Staph. aureus GB 2/1 (67%). SIGNIFICANCE AND IMPACT OF THE STUDY: It is concluded that biosurfactant represent suitable compounds that should be considered in developing future strategies to prevent the microbial colonization of silicone rubber voice prostheses. PMID- 16478487 TI - Changes to the phenotypic profile of Vibrio harveyi when infected with the Vibrio harveyi myovirus-like (VHML) bacteriophage. AB - AIMS: To determine if infection of Vibrio harveyi with the V. harveyi myovirus like (VHML) bacteriophage causes a change to the phenotypic profile of this species. METHODS AND RESULTS: Using 46 biochemical and metabolic tests, phenotypic profiles for noninfected V. harveyi and VHML infected V. harveyi were developed. Comparison of the infected and bacteriophage-infected strains of V. harveyi 645, 20 and 45 were found to have different test results for d-gluconate utilization, gamma-glutamyl transpeptidase and sulfatase activity, respectively. Using probabilistic identification, VHML infected and noninfected strains were identified as V. harveyi and had similar Willcox probability scores though the modal likelihood scores were reduced for VHML infected strains. One VHML infected strain, 642b, was misidentified as V. campbellii by phenotyping but not by PCR. It would appear that the phenotype of V. harveyi strains infected with VHML, are sufficiently altered that they occur at the margins of the known range of strain variation for V. harveyi. CONCLUSION: Infection of V. harveyi with VHML causes the phenotypic profile of the bacterium to change. This change reduces the modal likelihood score resulting in a poorer level of assurance for an identification of V. harveyi, especially in the natural host, strain 642. The bacteriophage VHML integrates into different sites in different strains of V. harveyi. SIGNIFICANCE AND IMPACT OF THE STUDY: The identification of V. harveyi as the causative agent of mortality in aquatic organisms is predominantly achieved through phenotyping. Since bacteriophages alter virulence in V. harveyi, understanding the effect they have on phenotype is important. PMID- 16478488 TI - Linking bacteriophage infection to quorum sensing signalling and bioluminescent bioreporter monitoring for direct detection of bacterial agents. AB - AIM: To incorporate into the lambda phage genome, a luxI-based acyl-homoserine lactone (AHL) synthase genetic construct and exploit the autoamplified power of quorum sensing to translate a phage infection event into a chemical signature detectable by a lux-based bioluminescent bioreporter, with focus towards facile detection of microbial pathogens. METHODS AND RESULTS: The luxI gene from Vibrio fischeri was inserted into the lambda phage genome to construct a model phage based biosensor system for the general detection of Escherichia coli. The AHL signalling molecules synthesized upon phage infection are detected by an AHL specific bioluminescent bioreporter based on the luxCDABE gene cassette of V. fischeri. The assay generates target-specific visible light signals with no requisite addition of extraneous substrate. This binary reporter system was able to autonomously respond to lambda phage infection events at target E. coli concentrations ranging from 1 x 10(8) to 1 CFU ml(-1) within 1.5-10.3 h, respectively, in pure culture. When assayed against artificially contaminated lettuce leaf washings, detection within an E. coli inoculum range from 1 x 10(8) to 130 CFU ml(-1) was achieved within 2.6-22.4 h, respectively. CONCLUSIONS: The initial feasibility of binary phage-based reporter assays indicates that quorum sensing can be used to translate a phage infection event into an autoamplified chemical signature. SIGNIFICANCE AND IMPACT OF STUDY: With further modification, binary phage-based reporter assays may be capable of rapidly and cost effectively detecting pathogenic agents at very low population densities. PMID- 16478489 TI - Determination of the amino acid residues required for the activity of the anti rhizobial peptide antibiotic trifolitoxin. AB - AIMS: The first aim was to determine those amino acid residues required for the biological activity of the potent peptide antibiotic, trifolitoxin (TFX). The second aim was to determine the concentrations of TFX1 and TFX2 that cause 50% inhibition of bacterial growth (Ki), the two predominant isomeric forms of TFX made by Rhizobium. METHODS AND RESULTS: Site-directed mutagenesis of tfxA was used to produce strains that made mutant TFX peptides. The mutant tfxA genes were placed on a vector and inserted in Rhizobium leguminosarum b. trifolii Tn54A112, a tfxA mutant of strain T24 that lacks trifolitoxin activity. Our standard bioassay was used to assess the activity of these mutants. TFX1 and TFX2 were purified by reverse phase chromatography. Several concentrations of each peptide were assayed for biological activity to determine Ki. The unmodified TFX peptide (DIGGSRQGCVA) was synthesized and was found to lack any biological activity. Four of the 11 amino acid residues in ribosomally synthesized, post-translationally modified peptide were required for TFX activity. These required amino acids include arginine (R37), glutamine (Q38), glycine (G39) and cysteine (C40). S36T and S36Y mutants showed reduced TFX activity. The numbering system is based on the 42-amino acid TfxA peptide that is post-translationally modified to form the active TFX peptide. The Ki of TFX2 was determined to be 10-fold lower than TFX1. CONCLUSIONS: The post-translational modifications of the TfxA peptide are required for biological activity. TFX2 is far more active than TFX1. SIGNIFICANCE AND IMPACT OF THE STUDY: The sequence of the TfxA peptide appears to have been optimized for maximum activity through the course of evolution. Even conservative changes to any of the amino acid residues required for activity results in a complete loss of activity. The understanding of the action of this peptide is critical for its proposed action as a control agent for crown gall disease. PMID- 16478490 TI - Combination of antagonistic yeasts with two food additives for control of brown rot caused by Monilinia fructicola on sweet cherry fruit. AB - AIMS: To evaluate beneficial effect of two food additives, ammonium molybdate (NH4-Mo) and sodium bicarbonate (NaBi), on antagonistic yeasts for control of brown rot caused by Monilinia fructicola in sweet cherry fruit under various storage conditions. The mechanisms of action by which food additives enhance the efficacy of antagonistic yeasts were also evaluated. METHODS AND RESULTS: Biocontrol activity of Pichia membranefaciens and Cryptococcus laurentii against brown rot in sweet cherry fruit was improved by addition of 5 mmol l(-1) NH4-Mo or 2% NaBi when stored in air at 20 and 0 degrees C, and in controlled atmosphere (CA) storage with 10% O2 + 10% CO2 at 0 degrees C. Population dynamics of P. membranefaciens in the wounds of fruit were inhibited by NH4-Mo at 20 degrees C after 1 day of incubation and growth of C. laurentii was inhibited by NH4-Mo at 0 degrees C in CA storage after 60 days. In contrast, NaBi did not significantly influence growth of the two yeasts in fruit wounds under various storage conditions except that the growth of P. membranefaciens was stimulated after storage for 45 days at 0 degrees C in CA storage. When used alone, the two additives showed effective control of brown rot in sweet cherry fruit and the efficacy was closely correlated with the concentrations used. The result of in vitro indicated that growth of M. fructicola was significantly inhibited by NH4 Mo and NaBi. CONCLUSION: Application of additives improved biocontrol of brown rot on sweet cherry fruit under various storage conditions. It is postulated that the enhancement of disease control is directly because of the inhibitory effects of additives on pathogen growth, and indirectly because of the relatively little influence of additives on the growth of antagonistic yeasts. SIGNIFICANCE AND IMPACT OF THE STUDY: The results obtained in this study suggest that an integration of NH4-Mo or NaBi with biocontrol agents has great potential in commercial management of postharvest diseases of fruit. PMID- 16478492 TI - Occurrence of foodborne pathogenic bacteria in retail prepackaged portions of marine fish in Spain. AB - AIMS: To survey the presence of indigenous and nonindigenous foodborne bacterial pathogens in displayed prepacked portions of fresh marine fish. METHODS AND RESULTS: A survey of 50 different samples of fresh marine fish (conger, swordfish, sole, grouper and whiting) was conducted over a period of 5 months. Trays of fillets and steaks were obtained at retail level and tested for foodborne bacterial pathogens. Vibrio cholerae and Salmonella were not detected. Two samples (4%) yielded Vibrio strains carrying a DNA fragment specific for Vibrio parahaemolyticus, but resulted negative to PCR amplification of the virulence-related tdh gene. Levels of motile Aeromonas ranging from 2.29 to 7.20 log CFU g(-1) were found in 31 (62%) samples. All fish portions were positive for the Aeromonas hlyA gene and 38 for both aerA and hlyA genes, which may contribute to diarrhoea-related virulence. The incidence of Listeria monocytogenes was 10%. Levels of Staphylococcus aureus lower than 2 log CFU g(-1) were found in 15 (30%) samples. Numbers of presumptive Clostridium perfringens ranging from 1.82 +/- 0.22 to 4.26 +/- 1.25 log CFU g(-1) were detected in 42 (84%) samples. Edwardsiella tarda was detected in two samples of grouper fillets. CONCLUSIONS: Displayed portions of raw fish carried bacteria that can cause foodborne disease. The risk posed by fresh fish when properly cooked is low, but high when destined to be consumed raw, undercooked or very lightly processed. SIGNIFICANCE AND IMPACT OF THE STUDY: This study revealed that raw fish sold in Spain could be a source of foodborne bacterial pathogens. Improvements in handling and processing are needed to minimize the prevalence of pathogenic bacteria. PMID- 16478491 TI - Genetic characterization of strains of Saccharomycescerevisiae responsible for 'refermentation' in Botrytis-affected wines. AB - AIMS: Saccharomyces cerevisiae is responsible for alcoholic fermentation of wines. However, some strains can also spoil sweet Botrytis-affected wines. Three 'refermentation' strains were isolated during maturation. Characterization of those strains in regards to their fingerprint, rDNA sequence and resistance to SO2, which constituted the main source of stress in Botrytis-affected wines, was carried out. METHODS AND RESULTS: Refermentation strains could be clearly discriminated by interdelta fingerprinting. However, they exhibited close relationships by karyotyping. A part of RDN1 locus sequence was examined by using PCR-RFLP and PCR-DGGE. The resistance of refermentation strains to SO2 was performed by using real time quantitative PCR focusing on SSU1 gene. CONCLUSIONS: Results suggested that refermentation strains were heterozygote in 26S rDNA and their ITS1-5.8S rDNA-ITS2 region sequence revealed relationships with 'flor' strains. As described in the literature for flor strain, two out of three refermentation strains constitutively developed a higher level of SSU1 expression than the reference strains, improving their putative tolerance to SO2. Therefore, refermentation strains of S. cerevisiae had developed many strategies to survive during maturing sweet wines. SIGNIFICANCE AND IMPACT OF THE STUDY: Singularities in rDNA sequence and SSU1 overexpression revealed a natural adaptation. Moreover, genomic relationship between flor and refermentation strains suggested that stress sources could induced selection of survivor strains. PMID- 16478493 TI - Factors influencing the composition of bacterial communities found at abandoned copper-tailings dumps. AB - AIMS: To study the effect that copper residues exert on bacterial communities and the ability of bacteria to colonize different microhabitats in abandoned tailing dumps. METHODS AND RESULTS: We used the terminal-restriction fragment length polymorphism technique, a culture-independent molecular approach based on PCR amplification of ribosomal genes, to compare the structure of the bacterial communities from samples taken at two nearby located abandoned tailing dumps found in the Mediterranean-climate area of central Chile. Our results show that elevated available copper content in tailings has a strong effect on the bacterial community composition, but that other factors like pH and organic matter content also play an important role in the structure of these communities. We also found that the number of abundant bacteria in these samples was significantly lower than in soils not exposed to metal pollution. CONCLUSIONS: In addition to bioavailable copper, bacterial communities found in copper-tailings dumps are also affected by several other environmental factors. SIGNIFICANCE AND IMPACT OF THE STUDY: This first report on environmental factors influencing microbial communities in copper-tailings dumps will help to devise appropriate restoration procedures in this type of polluted habitat. PMID- 16478494 TI - A novel bacteriocin, thuricin 17, produced by plant growth promoting rhizobacteria strain Bacillus thuringiensis NEB17: isolation and classification. AB - AIMS: The aim of this study was to identify and characterize a compound produced by the plant growth promoting bacterium, Bacillus thuringiensis non Bradyrhizobium Endophytic Bacterium 17. METHODS AND RESULTS: The bacterial peptide was analysed and purified via HPLC. Using the disk diffusion assay this peptide inhibited the growth of 16/19 B. thuringiensis strains, 4/4 Bacillus cereus strains, among others, as well as a Gram-negative strain Escherichia coli MM294 (pBS42). Both bactericidal and bacteristatic effects were observed on B. cereus ATCC 14579 and bactericidal effects were observed on B. thuringiensis ssp. thuringiensis Bt1267. The molecular weight of the peptide was estimated via SDS PAGE and confirmed with Matrix Assisted Laser Desorption Ionization Quadrapole Time of Flight mass spectrometry; its weight is 3162 Da. The peptide is biologically active after exposure to 100 degrees C for 15 min, and within the pH range 1.00-9.25. Its activity disappeared when treated with proteinase K and protease, but not with alpha-amylase or catalase. CONCLUSIONS: We conclude that this is the first report of a bacteriocin produced by a plant growth promoting rhizobacteria (B. thuringiensis) species and have named the bacteriocin thuricin 17. SIGNIFICANCE AND IMPACT OF THE STUDY: Our work has characterized a bacteriocin produced by a plant growth promoting bacterium. This strain is previously reported to increase soya bean nodulation. PMID- 16478495 TI - Effect of iron(III), humic acids and anthraquinone-2,6-disulfonate on biodegradation of cyclic nitramines by Clostridium sp. EDB2. AB - AIMS: To determine the biodegradation of cyclic nitramines by an anaerobic marine bacterium, Clostridium sp. EDB2, in the presence of Fe(III), humic acids (HA) and anthraquinone-2,6-disulfonate (AQDS). METHODS AND RESULTS: An obligate anaerobic bacterium, Clostridium sp. EDB2, degraded RDX and HMX, and produced similar product distribution including nitrite, methylenedinitramine, nitrous oxide, ammonium, formaldehyde, formic acid and carbon dioxide. Carbon (C) and nitrogen (N) mass balance for RDX products were 87% and 82%, respectively, and for HMX were 88% and 74%, respectively. Bacterial growth and biodegradation of RDX and HMX were stimulated in the presence of Fe(III), HA and AQDS suggesting that strain EDB2 utilized Fe(III), HA and AQDS as redox mediators to transfer electrons to cyclic nitramines. CONCLUSIONS: Strain EDB2 demonstrated a multidimensional approach to degrade RDX and HMX: first, direct degradation of the chemicals; second, indirect degradation by reducing Fe(III) to produce reactive-Fe(II); third, indirect degradation by reducing HA and AQDS which act as electron shuttles to transfer electrons to the cyclic nitramines. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study could be helpful in determining the fate of cyclic nitramine energetic chemicals in the environments rich in Fe(III) and HA. PMID- 16478496 TI - Detection of norovirus capsid proteins in faecal and food samples by a real time immuno-PCR method. AB - AIMS: To develop a sensitive real time immuno-polymerase chain reaction (rtI-PCR) method for detecting norovirus (NV) capsid protein in food samples. METHODS AND RESULTS: The viral antigens were captured by two polyclonal antisera against recombinant Norwalk viral-like particles (rNVLPs). Biotin-conjugated antibodies, avidin and biotin-conjugated DNA reporter were used to convert the protein signals into DNA signals. The reporter DNA was then amplified by addition of primers and PCR. A real time PCR method was used in order to perform a quantitative post-PCR analysis. One hundred rNVLPs (10 fg) and a NV sample containing 660 rNVLPs equivalent particle units (66 fg) could be detected by this method. CONCLUSION: The PCR inhibitors present in the food samples had minimal effect on antigen capture and were removed by multiple wash steps during the rtI PCR procedure. The sensitivity of rtI-PCR was >1000-fold higher than the standard enzyme-linked immunosorbent assay and approximately 10 times higher than reverse transcription PCR in detection of NV capsid protein in stool and food samples. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of a rtI-PCR method to detect NV in contaminated food samples without concentration or purification of the virus. PMID- 16478497 TI - Cyclosporine A from a nonpathogenic Fusarium oxysporum suppressing Sclerotinia sclerotiorum. AB - AIMS: To evaluate the antagonistic activity of Fusarium oxysporum nonpathogenic fungal strain S6 against the phytopathogenic fungus Sclerotinia sclerotiorum and to identify the antifungal compounds involved. METHODS AND RESULTS: The antagonistic activity of Fusarium oxysporum strain S6 was determined in vitro by dual cultures. The metabolite responsible for the activity was isolated by chromatographic techniques, purified and identified by spectroscopic methods as cyclosporine A. The antifungal activity against the pathogen was correlated with the presence of this metabolite by a dilution assay and then quantified. Cyclosporine A caused both growth inhibition and suppression of sclerotia formation. In a greenhouse assay, a significant increase in the number of surviving soybean (Glycine max) plants was observed when S. sclerotiorum and F. oxysporum (S6) were inoculated together when compared with plants inoculated with S. sclerotiorum alone. CONCLUSION: Fusarium oxysporum (S6) may be a good fungal biological control agent for S. sclerotiorum and cyclosporine A is the responsible metabolite involved in its antagonistic activity in vitro. SIGNIFICANCE AND IMPACT OF THE STUDY: Cyclosporine A has not been previously described as an inhibitor of S. sclerotiorum. Its minimum inhibitory concentration (MIC) of 0.1 microg disc(-1) makes it suitable to use as a biofungicide. In vivo experiments showed that F. oxysporum (S6) is a good candidate for the biocontrol of S. sclerotiorum in soybean. PMID- 16478498 TI - Effectiveness of autochthonous bacterium and mycorrhizal fungus on Trifolium growth, symbiotic development and soil enzymatic activities in Zn contaminated soil. AB - AIMS: This study investigates how autochthonous micro-organisms [bacterium and/or arbuscular mycorrhizal (AM) fungi] affected plant tolerance to Zn contamination. METHODS AND RESULTS: Zinc-adapted and -nonadapted Glomus mosseae strains protected the host plant against the detrimental effect of Zn (600 microg g(-1)). Zn-adapted bacteria increased root growth and N, P nutrition in plants colonized by adapted G. mosseae and decreased the specific absorption rate (SAR) of Cd, Cu, Mo or Fe in plants colonized by Zn-nonadapted G. mosseae. Symbiotic structures (nodule number and extraradical mycelium) were best developed in plants colonized by those Zn-adapted isolates that were the most effective in increasing plant Zn tolerance. The bacterium also increased the quantity and quality (metabolic characteristics) of mycorrhizal colonization, with the highest improvement for arbuscular vitality and activity. Inocula also enhanced soil enzymatic activities (dehydrogenase, beta-glucosidase and phosphatase) and indol acetic acid (IAA) accumulation, particularly in the rhizosphere of plants inoculated with Zn adapted isolates. CONCLUSIONS: Glomus mosseae strains have a different inherent potential for improving plant growth and nutrition in Zn-contaminated soil. The bacterium increased the potential of mycorrhizal mycelium as inoculum. SIGNIFICANCE AND IMPACT OF THE STUDY: Mycorrhizal performance, particularly that of the autochthonous strain, was increased by the bacterium and both contributed to better plant growth and establishment in Zn-contaminated soils. PMID- 16478499 TI - Classical and molecular analyses to characterize commercial dry yeasts used in wine fermentations. AB - AIMS: The aim of the work was to apply PCR-temperature gradient gel electrophoresis (PCR-TGGE) and restriction enzyme analysis (RE) assays to identify commercially available starters of Saccharomyces cerevisiae sensu stricto complex. METHODS AND RESULTS: To characterize an analysed pool of 62 active dry yeasts of different brands used in wine fermentation practices, classical microbiological tests were also performed as well as evaluation of contamination with lactic acid bacteria and non-Saccharomyces yeasts. PCR-TGGE and RE were used in order to provide fast and reliable methods to identify and differentiate enological yeasts. Proposed molecular methods enabled to identify particular strains within 36 h after colony isolation and directly from dry yeast suspension. CONCLUSIONS: The methods are highly recommended to obtain reliable results on yeast strain differentiation in a significantly shorter time if compared to classical fermentation tests. SIGNIFICANCE AND IMPACT OF THE STUDY: The obtaining of yeast strain differentiation in a short time and without plating is a good tool for a rapid discrimination among enological strains used as starters in enological practices. PMID- 16478500 TI - Proposal of a method for the genetic transformation of Gordonia jacobaea. AB - AIMS: Gordonia jacobaea is a recently isolated bacterial species with potential industrial application on account of its ability to store large quantities of trans-canthaxanthin. Its genetic manipulation is, however, difficult and cumbersome owing to the presence of mycolic acids in the cell wall and, especially, because of current lack of knowledge about its basic genetics. The present work describes a method for the genetic transformation of G. jacobaea. METHODS AND RESULTS: Gordonia jacobaea was grown in media supplemented with different glycine, penicillin G and isoniazid concentrations. The temperature, carbon source, growth phase and ultrasounds were analyzed for improving the method efficiency. The cells were finally transformed by electroporation. Finally, the method was applied to Brevibacteriumlactofermentum and Gordonia bronchialis. CONCLUSIONS: The growth of G. jacobaea in the presence of glycine and isoniazid is essential for obtaining electrocompetents cells. The temperature, growth phase and ultrasounds appeared as the main factors for increasing the transformation efficiency. The use of shuttle plasmids became necessary. The method described can be used with other Corynebacteria species. SIGNIFICANCE AND IMPACT OF THE STUDY: Because of the importance of the CNM group (Corynebacteria, Nocardia and Mycobacteria genera) in different areas such as industry, bioremediation improve the knowledge of their molecular mechanisms are becoming essential. The method described here improves the genetic manipulation of this group of bacteria. PMID- 16478501 TI - Pore formers promoted release of an antifungal drug from functionalized polyurethanes to inhibit Candida colonization. AB - AIMS: As a preventive strategy to inhibit fungal biofilm formation on medical devices, we planned experiments based on polyurethane loading with fluconazole plus pore-former agents in order to obtain a promoted release of the antifungal drug. METHODS AND RESULTS: Different functional groups including carboxyl, hydroxyl, primary and tertiary amino groups, were introduced in polyurethanes. Fluconazole was adsorbed in higher amounts by the most hydrophilic polymers and its release was influenced by the degree of polymer swelling in water. The entrapping in the polymer of polyethylenglycol as a pore former significantly improved the fluconazole release while the entrapping of the higher molecular weight porogen albumin resulted in a controlled drug release and in an improved antifungal activity over time. CONCLUSIONS: Among the tested in vitro models, best results were achieved with an hydrophobic polymer impregnated with 25% (w/w) albumin and fluconazole which inhibited Candida albicans growth and biofilm formation on polymeric surfaces up to 8 days. SIGNIFICANCE AND IMPACT OF THE STUDY: The combined entrapping in polymers of pore formers and an antifungal drug and the consequent controlled release over time is a novel, promising approach in the development of medical devices refractory to fungal colonization. PMID- 16478502 TI - Propionibacterium acnes. AB - Propionibacterium acnes, a common skin organism, is most notably recognized for its role in acne vulgaris. It also causes postoperative and device-related infections and has been associated with a number of other conditions such as sarcoidosis and synovitis, acne, pustulosis, hyperostosis and osteitis (SAPHO), although its precise role as a causative agent remains to be determined. Propionibacterium acnes produces a number of virulence factors and is well known for its inflammatory and immunomodulatory properties. Recent publication of the P. acnes genome should provide further insights into the pathogenic capabilities of the organism and potentially lead to the development of new therapies. PMID- 16478503 TI - Determination of survival, identity and stress resistance of probiotic bifidobacteria in bio-yoghurts. AB - AIMS: To determine the level of bifidobacteria in bio-yoghurts in the UK, identify the species, and compare the resistance of common Bifidobacterium spp. to acidity and oxidative stress. METHODS AND RESULTS: A storage trial of bio yoghurts was carried out to determine the level and survival of bifidobacteria. The 16S rRNA gene targeted PCR was used to identify the species. Acid tolerance was determined by introducing the organisms to pH-adjusted skimmed milk and enumerating during storage at 4 degrees C. Oxidative stress resistance was determined using the H(2)O(2) disc diffusion assay technique. Nine of 10 bio yoghurts contained bifidobacteria at levels >10(6) CFU g(-1) at the time of purchase. The viability of the organism decreased during storage and on expiry only five products retained viability >10(6) CFU g(-1) while two others were very close to the target population. Bifidobacterium animalis ssp. lactis showed superior survival abilities and stress tolerance compared with Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium bifidum, Bifidobacterium adolescentis and Bifidobacterium longum biotype infantis. CONCLUSIONS: Bifidobacterium animalis ssp. lactis, the only Bifidobacterium spp. found in bio yoghurts, had the greatest resistance to acidity and oxidative stress. SIGNIFICANCE AND IMPACT OF THE STUDY: The technological properties of B. animalis ssp. lactis make it suitable for inclusion in bio-yoghurts although its putative health benefits need further investigation. PMID- 16478504 TI - OmpC and OmpF are required for growth under hyperosmotic stress above pH 8 in Escherichia coli. AB - AIMS: To investigate the requirement of outer membrane porins for osmotic adaptation at alkaline pH in Escherichia coli. METHODS AND RESULTS: Escherichia coli mutants deficient in ompC, ompF and both genes were constructed and the growth of these mutants was observed at alkaline pH. The growth rate of the mutant deficient in both ompC and ompF was slower than that of the wild type and mutants deficient in one of these genes under hyperosmotic stress at pHs above 8.0. The decreased rate was recovered when a cloned ompC was introduced to the mutant, but the growth recovery with a cloned ompF was partial. Such growth diminution was not observed at pHs below 8.0. CONCLUSION: OmpC and OmpF were shown to participate in hyperosmotic adaptation at alkaline pH in E. coli. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first report to demonstrate that OmpC and OmpF are required for hyperosmotic adaptation at pHs above 8.0, but not below 8.0. PMID- 16478505 TI - New convenient defined media for [(35)S]methionine labelling and proteomic analyses of probiotic lactobacilli. AB - AIMS: To develop experimental conditions for efficient protein radiolabelling and two-dimensional (2D) polyacrylamide gel electrophoresis for investigation of stress proteomes of probiotic Lactobacillus spp. METHODS AND RESULTS: Three chemically defined media (CDM) optimized from a commercial medium supported rapid growth of the probiotic Lactobacillus rhamnosus E97800, Lactobacillus brevis ATCC 8287 and Lactobacillus reuteri E97849, and a broad range of other lactic acid bacteria. These CDM allowed efficient protein radiolabelling, requiring as little as 200 mul of logarithmic culture and pulse-chase labelling of 20 min to detect c. 300 distinct protein spots in a mini-scale 2D-gel. Proteins including DnaK, GroEL and ClpATPases were identified from the 2D-gels by immunoblotting. CONCLUSIONS: Radiolabelling coupled with 2D gel electrophoresis provides a sensitive means to monitor changes in protein synthesis rates in probistic lactobacilli. SIGNIFICANCE AND IMPACT OF THE STUDY: Efficient tools for proteomic analyses of probiotic Lactobacillus were developed and applied for stress response studies. PMID- 16478506 TI - Expression of a thermostable xylanase gene from Bacillus coagulans ST-6 in Lactococcus lactis. AB - AIMS: The aim of the study is to evaluate whether xylanase can be used as a potential reporter gene for cloning and expression studies in Lactococcus. METHODS AND RESULTS: The 750 bp xylanase gene was amplified and subcloned into the unique NheI restriction enzyme site of pMG36e and subsequently transformed into competent Escherichia coli XLI-blue MRF cells and Lactococcus lactis cells. Bacterial culture containing pMG36e-Xy has an enzyme activity of 390 microg xylose ml(-1) culture 30 min(-1), respectively, when compared with 40 microg xylose ml(-1) culture 30 min(-1) for the negative control (plasmidless strain). CONCLUSIONS: The thermostable xylanase gene was successfully expressed in both E. coli and L. lactis. The activity of xylanase can be easily detected by the formation of visible clearing zones around the transformed colonies on Remazol Brilliant Blue-Xylan (RBB-Xylan) agar media. However, there were some significant differences in the optimum growth temperature and plasmid stability in the new clones. SIGNIFICANCE AND IMPACT OF THE STUDY: The constructed reporter vector has the potential to be used as a reporter system for Lactococcus as well as E. coli, and it is an addition to the pool of lactococcal vector systems. PMID- 16478508 TI - 16S rDNA directed PCR primers and detection of methanogens in the bovine rumen. AB - AIMS: To assess the diversity of ruminal methanogens in a grazing cow, and develop PCR primers targeting the predominant methanogens. METHODS AND RESULTS: DNA was extracted from rumen contents collected from a cow grazing pasture. Archaeal 16S rRNA genes were amplified by PCR using two pairs of archaea-specific primers, and clone libraries prepared. Selected clones were sequenced. Phylogenetic analysis revealed that for one primer pair, most sequences clustered with Methanobrevibacter spp. whereas with the other primer pair most clustered with Methanosphaera stadtmanae. One sequence belonged to the Crenarcheota. PCR primers were designed to detect Msp. stadtmanae and differentiate between Mbb. ruminantium and Mbb. smithii and successfully tested. CONCLUSIONS: The ruminal methanogens included Mbb. ruminantium, Mbb. smithii, Mbb. thaueri and methanogens similar to Msp.stadtmanae. The study showed that apparent methanogen diversity can be affected by selectivity from the archaea-specific primers used to create clone libraries. SIGNIFICANCE AND IMPACT OF THE STUDY: This study revealed a greater diversity of ruminal methanogens in grazing cows than previously recognized. It also shows the need for care in interpreting methanogen diversity using PCR-based analyses. The new PCR primers will enable more information to be obtained on Msp. stadtmanae and Methanobrevibacter spp. in the rumen. PMID- 16478507 TI - Bacteriocins as alternative agents for control of multiresistant staphylococcal strains. AB - AIMS: To investigate the activity of seven staphylococcins, bacteriocins produced by staphylococci, against multiresistant Staphylococcus aureus and coagulase negative staphylococci (CNS) involved in human infections. METHODS AND RESULTS: Four bacteriocins produced by Staph. epidermidis (Pep5, epidermin, epilancin K7 and epicidin 280) and three produced by Staph. aureus (aureocins A70, A53 and 215FN) were tested. Sixteen Staph. aureus strains, including a representative strain of the endemic Brazilian methicillin-resistant clone (MRSA), and 57 CNS strains were used as indicators. Among the staphylococcins used, Pep5 was able to inhibit 77.2% of the CNS strains and 87.5% of the Staph. aureus strains tested, including the Brazilian MRSA endemic clone, responsible for a large number of hospital-acquired infections in Brazil. On the other hand, aureocin A53 and epidermin presented a high antagonistic activity only against the Staph. aureus strains, being able to inhibit, respectively, 87.5% and 81.3% of them, including also the Brazilian MRSA endemic clone. The remaining bacteriocins inhibited only a low percentage of the nosocomial staphylococcal strains tested. CONCLUSIONS: Aureocin A53 and epidermin have potential applications against MRSA, whereas Pep5 seems to be an attractive agent against both MRSA and CNS, including mupirocin resistant strains and the Brazilian endemic clone of MRSA, which is also found disseminated in other countries. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacteriocins may represent alternative agents to control important nosocomial pathogens. PMID- 16478509 TI - Differentiation of outer membrane proteins from Salmonellaenterica serotypes using Fourier transform infrared spectroscopy and chemometrics. AB - AIMS: To differentiate between outer membrane proteins (OMPs) from six Salmonellaenterica serotypes using a Fourier transform infrared (FTIR) spectroscopy method and chemometrics. METHODS AND RESULTS: The OMPs from Salmonella serotypes (Typhimurium, Enteritidis, Thomasville, Hadar, Seftenberg and Brandenburg) were isolated using a sarcosyl extraction method. OMP profiles on SDS-PAGE exhibited two or three bands between 48 and 54 kDa. Spectra of 10 microl of OMP preparations (5 mg ml(-1)) dried on a gold reflective slide were collected using 128 scans at 4 cm(-1) resolution and units of log (1/R) and analyzed using canonical variate analysis (CVA) and linear discriminant analysis (LDA). The CVA of Salmonella OMP spectra in the 1800-1500 cm(-1) region separated the serotypes and LDA provided a 100% correct classification. CONCLUSIONS: The use of a FTIR method combined with chemometrics provided better differentiation of Salmonella OMPs than the OMP pattern analysis by SDS-PAGE. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study to demonstrate that spectra of OMP extracts from Salmonella serotypes can be used for 100% correct classification of the serotypes studied. PMID- 16478510 TI - Phenotypical and genotypical characteristics of root-nodulating bacteria isolated from annual Medicago spp. in Soummam Valley (Algeria). AB - AIMS: In the framework of agro-pastoral system management using local annual medics coupled with their native root-nodulating bacteria to extend pasture zones, increase forage yields and improve ovine and bovine breeding in Algeria, we investigated diversity of rhizobia from annual Medicago spp. (Medicago arabica, Medicago polymorpha, Medicago minima and Medicago orbicularis). METHODS AND RESULTS: Ten nodulating-isolates were characterized by morphological, cultural, physiological and biochemical features, SDS-PAGE analysis and PCR-RFLP of 16S rDNA. The results show some degree of genetic diversity among the isolates; three can be affiliated to Sinorhizobium meliloti, one to Rhizobium galegae and six were separate. CONCLUSIONS: Local annual medics would have a high degree of specificity in their symbiotic interaction. Furthermore, our results support the presence of Rh. galegae in the Mediterranean region. SIGNIFICANCE AND IMPACT OF THE STUDY: This work is a preliminary step towards selection of efficient symbiotic Medicago-rhizobia to develop inoculants for management of agro-pastoral systems using local annual medics in Algeria. PMID- 16478511 TI - Purification and molecular characterization of subtilisin-like alkaline protease BPP-A from Bacillus pumilus strain MS-1. AB - AIMS: The present study was conducted by screening zein-degrading bacteria in an attempt to obtain zein-degrading protease. METHODS AND RESULTS: Soil bacteria were screened by formation of a clear zone on zein plates. Characterization of a zein-degrading bacterium indicated a taxonomic affiliation to Bacillus pumilus, and was named MS-1 strain. The strain produced two different types of extracellular proteases, BPP-A and BPP-B. In this study, we purified and characterized BPP-A because it exhibited a higher ability to hydrolyze zein than BPP-B. When casein was used as the substrate, the optimal pH for BPP-A was 11.0. In BPP-A, zein was better substrate than casein at pH 13.0, whereas casein was better one than zein at pH 11.0. The bppA gene encoded a 383-amino acid pre-pro form of BPP-A, and mature BPP-A contained 275 amino acid residues. It was concluded that BPP-A belonged to the subtilisin family. CONCLUSION: A zein degrading bacterium assigned to B. pumilus produced two different types of extracellular proteases, BPP-A and BPP-B. BPP-A exhibited an ability to hydrolyze zein in an extreme alkaline condition. SIGNIFICANCE AND IMPACT OF THE STUDY: This is a first report on screening for zein-degrading micro-organisms. The subtilisin like protease BPP-A is possible to utilize as an industrial enzyme for the production of zein hydrolysates. PMID- 16478512 TI - Survival of inoculated Saccharomyces cerevisiae strain on wine grapes during two vintages. AB - AIMS: To investigate the influence of a specific ecological niche, the wine grape, on the survival and development of Saccharomyces cerevisiae. METHODS AND RESULTS: A strain with a rare phenotype was sprayed onto the grape surfaces and monitored through two vintages using a specific indicative medium and analysing the internal transcribed spacer regions in the 5.8S rDNA. During the ripening process, there was a progressive colonization of the surface of the undamaged and damaged grapes by epiphytic yeasts, up to the time of harvest. The damaged wine grapes showed a much greater epiphytic yeast population. However, the inoculated S. cerevisiae strain showed a scarce persistence on both undamaged and damaged wine grapes, and the damaged grapes did not appear to improve the grape surface colonization of this strain. CONCLUSIONS: Results indicated that wine grape is not a favourable ecological niche for the development and colonization of S. cerevisiae species. SIGNIFICANCE AND IMPACT OF THE STUDY: Results of this work are further evidence that S. cerevisiae is not specifically associated with natural environments such as damaged and undamaged wine grapes. PMID- 16478513 TI - Plasmids from Lactobacillus helveticus: distribution and diversity among natural isolates. AB - AIMS: To investigate the distribution and the level of diversity of extrachromosomal molecules in Lactobacillus helveticus strains in relation to their different ecological niches. METHODS AND RESULTS: The plasmid profile of 22 Lact. helveticus strains, isolated from five different Italian cheeses, was determined. Among the tested strains, there was a variable presence of plasmids: eight plasmid-free strains and the remaining with several plasmids that could be differentiated on the basis of number and molecular weight. The profiles showed between one and five plasmid bands, which size ranged between 2.3 and 31 kb. Four of these plasmids were further analyzed by restriction digestion and compared with the plasmids from Lact. helveticus ATCC 15009(T). Analyses and comparison of their primary structures and hybridization experiments revealed the presence of different DNA homology groups. CONCLUSIONS: This study indicates that within Lact. helveticus species, there is a high degree of variability in relation to the presence of plasmid molecules. Moreover, the structural diversity found among some of these plasmids allows to hypothesize the presence of different evolutionary lineages. SIGNIFICANCE AND IMPACT OF THE STUDY: Studies on plasmid distribution and diversity should be considered as an essential component in a continuing effort to explore microbial diversity as well as to understand the real role of plasmids in the flow of genetic information in natural bacterial communities. PMID- 16478515 TI - Application of an Escherichia coli green fluorescent protein-based lysine biosensor under nonsterile conditions and autofluorescence background. AB - AIMS: To examine the utility of an Escherichia coli green fluorescent protein (GFP) containing biosensor for quantification of bioavailable lysine in selected feed samples under nonsterile conditions and to estimate the background fluorescence of analyzed feed samples and evaluate the risk of confounding GFP emission from the lysine assay organism. METHODS AND RESULTS: Escherichia coli lysine auxotroph GFP based biosensor was used to determine the percentage of bioavailable lysine in two samples of soybean-, cottonseed-, and meat and bone meal under nonsterile conditions. The fluorescence emitted by GFP was successfully measured using a spectrofluorimeter to monitor bacterial growth response to protein-derived lysine and lysine containing small peptides. The autofluorescence of analyzed feed samples at different concentrations could also be estimated. CONCLUSIONS: When feed protein concentrations are decreased, autofluorescence interference can be avoided. SIGNIFICANCE: The E. coli lysine auxotroph GFP-based biosensor can successfully be used for the determination of bioavailable lysine in these selected animal feed proteins under nonsterile conditions. IMPACT OF THE STUDY: E. coli GFP biosensor for lysine has potential for routine application in animal feeds. PMID- 16478514 TI - Isolation, characterization of bacteriophages specific to Microlunatus phosphovorus and their application for rapid host detection. AB - AIMS: To isolate and characterize lytic-bacteriophages specific to Microlunatus phosphovorus, and prepare fluorescently labelled phages (FLPs) for the rapid detection of the host bacterium in activated sludge. METHODS AND RESULTS: Isolation of bacteriophages lytic to M. phosphovorus was attempted by applying supernatants of activated sludge processes on the lawn of M. phosphovorus JCM9379 for plaque formation. Thirteen bacteriophage isolates were obtained. The restriction fragment length polymorphism analysis distinguished them into two different bacteriophages designated as phiMP1 and phiMP2. They were found to possess double-stranded DNA and host specificity. Morphological observations were done by electron microscopy. The bacteriophage particles stained by SYBR Green I was shown to be applicable to detect their host bacterial cells mixed with activated sludge. CONCLUSIONS: Two M. phosphovorus-specific bacteriophages were isolated and classified as Siphoviridae. FLPs of them were prepared, and successfully applied to detect the host bacterium added into the activated sludge. SIGNIFICANCE AND IMPACT OF THE STUDY: At least some of bacteria in activated sludge are susceptible to their related bacteriophages. Bacteriophages lytic to activated sludge bacteria could be affecting the bacterial population in activated sludge. The FLPs could be used for the easy-rapid detection of their host bacterium in activated sludge. PMID- 16478516 TI - Effect of carbohydrates on the survival of Lactobacillus helveticus during vacuum drying. AB - AIMS: To assess four carbohydrates for the protective effect against Lactobacillus helveticus cells inactivation during vacuum drying, and to study the effect of selected carbohydrate on changes of inactivation kinetics. METHODS AND RESULTS: Early stationary phase L. helveticus cells grown in MRS media were recovered from fermentation broth, washed with PBS buffer (pH 7.0), and then mixed with different concentrations of four carbohydrates, namely lactose, sorbitol, inulin, and xanthan gum. Cells were dried in a vacuum drier at 100 mbar, 43 degrees C for 12 h. Only cells with 1% sorbitol addition showed higher survival (18%) over cells without added carbohydrate (8%). Using in situ microbalance technique whereby cell weight during vacuum drying was continuously monitored via precision balances integrated into the vacuum chamber, drying and inactivation kinetics of cells and cells mixed with sorbitol were established. CONCLUSION: Survival of L. helveticus during the vacuum drying could be improved by the addition of optimal concentration of 1% sorbitol. Addition of sorbitol did not cause drastic changes in drying rate, water content and water activity of samples. The protection mechanisms of sorbitol seemed not to be due to a direct physical effect, which could be related to drying rate. SIGNIFICANCE AND IMPACT OF THE STUDY: The increase in survival of cells after vacuum drying by the addition of a protective carbohydrate may provide an alternative mean to preserve starter cultures at a higher level of activity. PMID- 16478517 TI - Characteristics of bacteria showing high denitrification activity in saline wastewater. AB - AIMS: Denitrification efficiency at 10% salinity was compared with that at 2% salinity. The characteristics of bacterial strains isolated from the denitrification system, where an improvement of denitrification efficiency was observed at a high salinity were investigated. METHODS AND RESULTS: Two continuous feeding denitrification systems for saline solutions of 2% and 10% salinity, were operated. Denitrification efficiency at 10% salinity was higher than that at 2% salinity. The bacterial strains were isolated using the trypticase soy agar (TSA) medium at 30 degrees C. The phylogenetic analysis of 16S rRNA gene sequences of isolates indicated that halophilic species were predominant at 10% salinity. CONCLUSIONS: The improvement of denitrification efficiency at a high salinity was demonstrated. The strains isolated from the denitrifying system with 10% salinity were halophilic bacteria, Halomonas sp. and Marinobacter sp., suggesting that these bacteria show a high denitrifying activity at 10% salinity. SIGNIFICANCE AND IMPACT OF THE STUDY: The long-term acclimated sludge used in this study resulted in high denitrification performance at a high salinity, indicating that the design of a high-performance denitrification system for saline wastewater will be possible. PMID- 16478519 TI - Evidence of an antilisterial factor induced by wounding of iceberg lettuce tissues. AB - AIMS: To examine the influence of wound-associated reactions in cut iceberg lettuce (Lactuca sativa L.) tissues on the fate of Listeria monocytogenes. METHODS AND RESULTS: Aqueous extracts prepared from shredded iceberg lettuce before and after storage in high oxygen permeability film were inoculated with L. monocytogenes. Listeria monocytogenes grew in extracts prepared from fresh lettuce. In contrast, inhibition ranging from arrested growth to a decline in cell viability was observed in extracts prepared from samples stored for 1-3 days. Similar behaviour was evident in lettuce shreds inoculated with 10(5) CFU g(-1)L. monocytogenes immediately after processing or after 3 days in storage. Heat treatment of the cut tissues at 47 degrees C for 3 min before storage diminished the inhibitory effect. CONCLUSIONS: The results provided evidence that an antilisterial factor or factors are released by wounded iceberg lettuce tissues. Antilisterial activity was mitigated by heat treatment of the lettuce. SIGNIFICANCE AND IMPACT OF STUDY: This study indicates that intrinsic factors associated with plant metabolism could play a significant role in the ecology of human pathogens in packaged horticultural products. PMID- 16478518 TI - Development of a multiplex-PCR for rapid detection of the enteric pathogens Lawsonia intracellularis, Brachyspira hyodysenteriae, and Brachyspira pilosicoli in porcine faeces. AB - AIMS: To develop an assay to simultaneously detect Lawsonia intracellularis, Brachyspira hyodysenteriae and Brachyspira pilosicoli in pig faeces. METHODS AND RESULTS: A multiplex-polymerase chain reaction (M-PCR) was designed to amplify a 655-base pair (bp) portion of the L. intracellularis 16S rRNA gene, a 354-bp portion of the B. hyodysenteriae NADH oxidase gene, and a 823-bp portion of the B. pilosicoli 16S rRNA gene. Specificity was assessed using 80 strains of Brachyspira spp. and 30 other enteric bacteria. Bacterial DNA was extracted from faeces using the QIAamp DNA Stool Mini Kit. The M-PCR was tested in parallel with culture and/or PCR on 192 faecal samples from eight piggeries. Faeces also were seeded with known cell concentrations of the three pathogenic species, and the limits of detection of the M-PCR tested. The M-PCR was specific, with limits of detection of 10(2)-10(3) cells of the respective species per gram of faeces. CONCLUSIONS: The M-PCR is a rapid, sensitive and specific test for detecting three important enteric bacterial pathogens of pigs. SIGNIFICANCE AND IMPACT OF THE STUDY: The availability of a new diagnostic M-PCR will allow rapid detection and control of three key porcine enteric pathogens. PMID- 16478520 TI - Bioavailable cadmium during the bioremediation of phenanthrene-contaminated soils using the diffusive gradients in thin-film technique. AB - AIMS: To study the impact of fungal bioremediation of phenanthrene on trace cadmium solid-solution fluxes and solution phase concentration. METHODS AND RESULTS: The bioremediation of phenanthrene in soils was performed using the fungus Penicillium frequentans. Metal behaviour was evaluated by the techniques of diffusive gradient in thin-films (DGT) and filtration. Fluxes of cadmium (Cd) show a significant (P < 0.002) increase after the start of bioremediation, indicating that the bioremediation process itself releases significant amount of Cd into solution from the soil solid-phase. Unlike DGT devices, the solution concentration from filtration shows a clear bimodal distribution. We postulate that the initial action of the fungi is most likely to breakdown the surface of the solid phase to smaller, 'solution-phase' material (<0.45 microm) leading to a peak in Cd concentration in solution. CONCLUSIONS: Phenanthrene removal from soils by bioremediation ironically results in the mobilization of another toxic pollutant (Cd). SIGNIFICANCE AND IMPACT OF THE STUDY: Bioremediation of organic pollutants in contaminated soil will likely lead to large increases in the mobilization of toxic metals, increasing metal bio-uptake and incorporation into the wider food chain. Bioremediation strategies need to account for this behaviour and further research is required both to understand the generality of this behaviour and the operative mechanisms. PMID- 16478521 TI - Plasmid-mediated bacteriocin production by Shigella flexneri isolated from dysenteric diarrhoea and their transformation into Escherichia coli. AB - AIMS: To determine the production of bacteriocin by Shigella flexneri strains, to relate their production to the presence of dysenteric diarrhoea and to asses the genetic determination of the bacteriocin. METHODS AND RESULTS: One hundred and sixteen strains of Sh. flexneri were isolated from patients with diarrhoea and 49 of them produced bacteriocin active against several Escherichia coli and abacteriocinogenic Sh. flexneri strains. The extrachromosomal DNA isolated from bacteriocinogenic Sh. flexneri strains were used as a substrate to transform E. coli HB-101 cells by means of electroporation. CONCLUSIONS: Only the Sh. flexneri strains isolated from dysenteric diarrhoea produced bacteriocin. It was demonstrated that a plasmid of approx. 3 kb was responsible for the genetic determination of these anti-bacterial substances. SIGNIFICANCE AND IMPACT OF THE STUDY: A 3-kb plasmid that harboured information for the production of bacteriocin by Sh. flexneri strains was described. The production of this bacteriocin may be related to dysenteric diarrhoea produced by these bacterial strains. PMID- 16478523 TI - The BH3-only protein Puma is both necessary and sufficient for neuronal apoptosis induced by DNA damage in sympathetic neurons. AB - DNA damage activates apoptosis in several neuronal populations and is an important component of neuropathological conditions. While it is well established that neuronal apoptosis, induced by DNA damage, is dependent on the key cell death regulators p53 and Bax, it is unknown which proteins link the p53 signal to Bax. Using rat sympathetic neurons as an in vitro model of neuronal apoptosis, we show that cytosine arabinoside is a DNA damaging drug that induces the expression of the BH3-only pro-apoptotic genes Noxa, Puma and Bim. Increased expression occurred after p53 activation, measured by its phosphorylation at serine 15, but prior to the conformational change of Bax at the mitochondria, cytochrome c (cyt c) release and apoptosis. Hence Noxa, Puma and Bim could potentially link p53 to Bax. We directly tested this hypothesis by the use of nullizygous mice. We show that Puma, but not Bim or Noxa, is a crucial mediator of DNA damage-induced neuronal apoptosis. Despite the powerful pro-apoptotic effects of overexpressed Puma in Bax-expressing neurons, Bax nullizygous neurons were resistant to Puma induced death. Therefore, Puma provides the critical link between p53 and Bax, and is both necessary and sufficient to mediate DNA damage-induced apoptosis of sympathetic neurons. PMID- 16478524 TI - A role for mixed lineage kinases in granule cell apoptosis induced by cytoskeletal disruption. AB - Microtubule disruption by colchicine induces apoptosis in selected neuronal populations. However, little is known about the upstream death signalling events mediating the neurotoxicity. We investigated first whether colchicine-induced granule cell apoptosis activates the c-Jun N-terminal kinase (JNK) pathway. Cultured murine cerebellar granule cells were exposed to 1 microm colchicine for 24 h. Activation of the JNK pathway was detected by western blotting as well as immunocytochemistry using antibodies against phospho-c-Jun (p-c-Jun). Next, adult male rats were injected intracerebroventricularly with colchicine (10 microg), and JNK pathway activation in dentate granule cells (DGCs) was detected by antibodies against p-c-Jun. The second part of the study tested the involvement of mixed lineage kinases (MLK) as upstream activators of the JNK pathway in colchicine toxicity, using CEP-1347, a potent MLK inhibitor. In vitro, significant inhibition of the JNK pathway, activated by colchicine, was achieved by 100-300 nm CEP-1347, which blocked both activation of cell death proteases and apoptosis. Moreover, CEP-1347 markedly delayed neurite fragmentation and cell degeneration. In vivo, CEP-1347 (1 mg/kg) significantly prevented p-c-jun increase following injection of colchicine, and enhanced survival of DGCs. We conclude that colchicine-induced neuronal apoptosis involves the JNK/MLK pathway, and that protection of granule cells can be achieved by MLK inhibition. PMID- 16478526 TI - The interaction between dopamine D2-like and beta-adrenergic receptors in the prefrontal cortex is altered by mood-stabilizing agents. AB - Several studies have suggested the involvement of biogenic monoaminergic neurotransmission in bipolar disorder and in the therapy for this disease. In this study, the effects of the mood-stabilizing drugs lithium, carbamazepine or valproate on the dopaminergic and adrenergic systems, particularly on D2-like and beta-adrenergic receptors, were studied both in cultured rat cortical neurones and in rat prefrontal cortex. In vitro and in vivo data showed that stimulation of beta-adrenergic receptors with isoproterenol increased cyclic adenosine monophosphate (cAMP) levels and this effect was significantly inhibited by lithium, carbamazepine or valproate. The activation of dopamine D2-like receptors with quinpirole decreased the isoproterenol-induced rise in cAMP in control conditions. This inhibition was observed in vivo after chronic treatment of the rats with carbamazepine or valproate, but not after treatment with lithium or in cultured rat cortical neurones after 48 h exposure to the three mood stabilizers. Dopamine D2 and beta1-adrenergic receptors were found to be co-localized in prefrontal cortical cells, as determined by immunohistochemistry, but western blot experiments revealed that receptor levels were differentially affected by treatment with the three mood stabilizers. These data show that mood stabilizers affect D2 receptor-mediated regulation of beta-adrenergic signalling and that each drug acts by a unique mechanism. PMID- 16478525 TI - Mutations in amyloid precursor protein and presenilin-1 genes increase the basal oxidative stress in murine neuronal cells and lead to increased sensitivity to oxidative stress mediated by amyloid beta-peptide (1-42), HO and kainic acid: implications for Alzheimer's disease. AB - Oxidative stress is observed in Alzheimer's disease (AD) brain, including protein oxidation and lipid peroxidation. One of the major pathological hallmarks of AD is the brain deposition of amyloid beta-peptide (Abeta). This 42-mer peptide is derived from the beta-amyloid precursor protein (APP) and is associated with oxidative stress in vitro and in vivo. Mutations in the PS-1 and APP genes, which increase production of the highly amyloidogenic amyloid beta-peptide (Abeta42), are the major causes of early onset familial AD. Several lines of evidence suggest that enhanced oxidative stress, inflammation, and apoptosis play important roles in the pathogenesis of AD. In the present study, primary neuronal cultures from knock-in mice expressing mutant human PS-1 and APP were compared with those from wild-type mice, in the presence or absence of various oxidizing agents, viz, Abeta(1-42), H2O2 and kainic acid (KA). APP/PS-1 double mutant neurons displayed a significant basal increase in oxidative stress as measured by protein oxidation, lipid peroxidation, and 3-nitrotyrosine when compared with the wild-type neurons (p < 0.0005). Elevated levels of human APP, PS-1 and Abeta(1 42) were found in APP/PS-1 cultures compared with wild-type neurons. APP/PS-1 double mutant neuron cultures exhibited increased vulnerability to oxidative stress, mitochondrial dysfunction and apoptosis induced by Abeta(1-42), H2O2 and KA compared with wild-type neuronal cultures. The results are consonant with the hypothesis that Abeta(1-42)-associated oxidative stress and increased vulnerability to oxidative stress may contribute significantly to neuronal apoptosis and death in familial early onset AD. PMID- 16478527 TI - Neural mitochondrial Ca2+ capacity impairment precedes the onset of motor symptoms in G93A Cu/Zn-superoxide dismutase mutant mice. AB - Mitochondrial respiratory chain dysfunction, impaired intracellular Ca2+ homeostasis and activation of the mitochondrial apoptotic pathway are pathological hallmarks in animal and cellular models of familial amyotrophic lateral sclerosis associated with Cu/Zn-superoxide dismutase mutations. Although intracellular Ca2+ homeostasis is thought to be intimately associated with mitochondrial functions, the temporal and causal correlation between mitochondrial Ca2+ uptake dysfunction and motor neuron death in familial amyotrophic lateral sclerosis remains to be established. We investigated mitochondrial Ca2+ handling in isolated brain, spinal cord and liver of mutant Cu/Zn-superoxide dismutase transgenic mice at different disease stages. In G93A mutant transgenic mice, we found a significant decrease in mitochondrial Ca2+ loading capacity in brain and spinal cord, as compared with age-matched controls, very early on in the course of the disease, long before the onset of motor weakness and massive neuronal death. Ca2+ loading capacity was not significantly changed in liver G93A mitochondria. We also confirmed Ca2+ capacity impairment in spinal cord mitochondria from a different line of mice expressing G85R mutant Cu/Zn-superoxide dismutase. In excitable cells, such as motor neurons, mitochondria play an important role in handling rapid cytosolic Ca2+ transients. Thus, mitochondrial dysfunction and Ca2+-mediated excitotoxicity are likely to be interconnected mechanisms that contribute to neuronal degeneration in familial amyotrophic lateral sclerosis. PMID- 16478529 TI - Histamine protects against NMDA-induced necrosis in cultured cortical neurons through H receptor/cyclic AMP/protein kinase A and H receptor/GABA release pathways. AB - Using histamine and the H3 receptor antagonist thioperamide, the roles of histamine receptors in NMDA-induced necrosis were investigated in rat cultured cortical neurons. Within 3 h of intense NMDA insult, most neurons died by necrosis. Histamine reversed the neurotoxicity in a concentration-dependent manner and showed peak protection at a concentration of 10(-7) m. This protection was antagonized by the H2 receptor antagonists cimetidine and zolantidine but not by the H1 receptor antagonists pyrilamine and diphenhydramine. In addition, the selective H2 receptor agonist amthamine mimicked the protection by histamine. This action was prevented by cimetidine but not by pyrilamine. 8-Bromo-cAMP also mimicked the effect of histamine. In contrast, both the adenylyl cyclase inhibitor 9-(tetrahydro-2-furanyl)-9H-purine-6-amine and the cAMP-dependent protein kinase inhibitor N-[2-(p-bromocinnamylamino) ethyl]-5 isoquinolinesulfonamide reversed the protection by histamine. Thioperamide also attenuated NMDA-induced excitotoxicity, which was reversed by the H3 receptor agonist (R)-alpha-methylhistamine but not by pyrilamine and cimetidine. In addition, the protection by thioperamide was inhibited by the GABA(A) receptor antagonists picrotoxin and bicuculline. Further study demonstrated that the protection by thioperamide was due to increased GABA release in NMDA-stimulated samples. These results indicate that not only the H2 receptor/cAMP/cAMP-dependent protein kinase pathway but also the H3 receptor/GABA release pathway can attenuate NMDA-induced neurotoxicity. PMID- 16478530 TI - Shift in the ratio of three-repeat tau and four-repeat tau mRNAs in individual cholinergic basal forebrain neurons in mild cognitive impairment and Alzheimer's disease. AB - Molecular mechanisms underlying tauopathy remain undetermined. In the current study, single cell gene expression profiling was coupled with custom-designed cDNA array analysis to evaluate tau expression and other cytoskeletal elements within individual neuronal populations in patients with no cognitive impairment (NCI), mild cognitive impairment (MCI), and Alzheimer's disease (AD). Results revealed a shift in the ratio of three-repeat tau (3Rtau) to four-repeat tau (4Rtau) mRNAs within individual human cholinergic basal forebrain (CBF) neurons within nucleus basalis (NB) and CA1 hippocampal neurons during the progression of AD, but not during normal aging. A shift in 3Rtau to 4Rtau may precipitate a cascade of events in the selective vulnerability of neurons, ultimately leading to frank neurofibrillary tangle (NFT) formation in tauopathies including AD. PMID- 16478528 TI - Regulation of taurine transport in rat hippocampal neurons by hypo-osmotic swelling. AB - Taurine, an important mediator of cellular volume regulation in the central nervous system, is accumulated into neurons and glia by means of a highly specific sodium-dependent membrane transporter. During hyperosmotic cell shrinkage, net cellular taurine content increases as taurine transporter activity is enhanced via elevated gene expression of the transporter protein. In hypo osmotic conditions, taurine is rapidly lost from cells by means of taurine conducting membrane channels. We reasoned that changes in taurine transporter activity also might accompany cell swelling to minimize re-accumulation of taurine from the extracellular space. Thus, we determined the kinetic and pharmacological characteristics of neuronal taurine transport and the response to osmotic swelling. Accumulation of radioactive taurine is strongly temperature dependent and occurs via saturable and non-saturable pathways. At concentrations of taurine expected in extracellular fluid in vivo, 98% of taurine accumulation would occur via the saturable pathway. This pathway obeys Michaelis-Menten kinetics with a Km of 30.0 +/- 8.8 microm (mean +/- SE) and Jmax of 2.1 +/- 0.2 nmol/mg protein min. The saturable pathway is dependent on extracellular sodium with an effective binding constant of 80.0 +/- 3.1 mm and a Hill coefficient of 2.1 +/- 0.1. This pathway is inhibited by structural analogues of taurine and by the anion channel inhibitors, 4,4'-diisothiocyanostilbene-2, 2'-disulfonic acid (DIDS) and 5-nitro-2-(3 phenylpropylamino) benzoic acid (NPPB). NPPB, but not DIDS, also reduces the ATP content of the cell cultures. Osmotic swelling at constant extracellular sodium concentration reduces the Jmax of the saturable transport pathway by approximately 48%, increases Kdiff for the non-saturable pathway by 77%, but has no effect on cellular ATP content. These changes in taurine transport occurring in swollen neurons in vivo would contribute to net reduction of taurine content and resulting volume regulation. PMID- 16478531 TI - Gene expression profiling reveals that peripheral nerve regeneration is a consequence of both novel injury-dependent and reactivated developmental processes. AB - One of the most striking features of the injured mature peripheral nervous system is the ability to regenerate. The lesioned peripheral nervous system displays stereotypic histopathological reactions indicating the activation of a co ordinated lesion-induced gene expression programme. Previous research has already identified molecular components of this axonal switch from a mature transmitting to a regenerative growth mode. The observed alterations in gene expression within the lesioned distal nerve stump were largely attributed to recapitulated developmental processes. However, to our knowledge, this hypothesis has not been proven systematically. Most of the stereotypic molecular and cellular reactions during nerve development and repair can be assigned to specific time windows. Consequently, we have compared gene expression profiles of both paradigms at six different time-points each by means of cDNA array hybridization. Our data identified injury-specific molecular reactions and revealed to what extent developmental mechanisms are reactivated in response to nerve lesion. Ninety-one genes (47% of the regeneration-associated genes) were found to be significantly regulated in both paradigms, suggesting that regeneration only partially recapitulates development and that approximately half of the regulated genes are part of a regeneration-dependent programme. Interestingly, mainly genes encoding signal transducers or factors involved in processes such as cell death, immune response, transport and transcriptional regulation showed injury-specific gene expression. PMID- 16478532 TI - Absence of synapsin I and II is accompanied by decreases in vesicular transport of specific neurotransmitters. AB - Studies of synapsin-deficient mice have shown decreases in the number of synaptic vesicles but knowledge about the consequences of this decrease, and which classes of vesicles are being affected, has been lacking. In this study, glutamatergic, GABAergic and dopaminergic transport has been analysed in animals where the genes encoding synapsin I and II were inactivated. The levels of the vesicular glutamate transporter (VGLUT) 1, VGLUT2 and the vesicular GABA transporter (VGAT) were decreased by approximately 40% in adult forebrain from mice devoid of synapsin I and II, while vesicular monoamine transporter (VMAT) 2 and VGLUT3 were present in unchanged amounts compared with wild-type mice. Functional studies on synaptic vesicles showed that the vesicular uptake of glutamate and GABA was decreased by 41 and 23%, respectively, while uptake of dopamine was unaffected by the lack of synapsin I and II. Double-labelling studies showed that VGLUT1 and VGLUT2 colocalized fully with synapsin I and/or II in the hippocampus and neostriatum, respectively. VGAT showed partial colocalization, while VGLUT3 and VMAT2 did not colocalize with either synapsin I or II in the brain areas studied. In conclusion, distinct vesicular transporters show a variable degree of colocalization with synapsin proteins and, hence, distinct sensitivities to inactivation of the genes encoding synapsin I and II. PMID- 16478534 TI - Zebrafish transgenic Enhancer TRAP line database (ZETRAP). AB - BACKGROUND: The zebrafish, Danio rerio, is used as a model organism to study vertebrate genetics and development. An effective enhancer trap (ET) in zebrafish using the Tol2 transposon has been demonstrated. This approach could be used to study embryogenesis of a vertebrate species in real time and with high resolution. DESCRIPTION: The information gathered during the course of systematic investigation of many ET transgenic lines have been collected and compiled in the form of an online database--the Zebrafish Enhancer TRAP lines database (ZETRAP). CONCLUSION: ZETRAP is a web-based system that provides data and information to the scientific community about the developmental, genetic and genomic aspects of transgenic zebrafish lines obtained using Tol2 transposon-mediated transgenesis. The current version (version 1.0) contains description of 27 ET lines that express EGFP in various organs and tissues, for example, heart, brain, notochord, gut, etc. It also includes information on insertion sites of the Tol2 transposon in these lines. PMID- 16478535 TI - Advising overweight persons about diet and physical activity in primary health care: Lithuanian health behaviour monitoring study. AB - BACKGROUND: Obesity is a globally spreading health problem. Behavioural interventions aimed at modifying dietary habits and physical activity patterns are essential in prevention and management of obesity. General practitioners (GP) have a unique opportunity to counsel overweight patients on weight control. The purpose of the study was to assess the level of giving advice on diet and physical activity by GPs using the data of Lithuanian health behaviour monitoring among adult population. METHODS: Data from cross-sectional postal surveys of 2000, 2002 and 2004 were analysed. Nationally representative random samples were drawn from the population register. Each sample consisted of 3000 persons aged 20 64 years. The response rates were 74.4% in 2000, 63.4% in 2002 and 61.7% in 2004. Self-reported body weight and height were used to calculate body mass index (BMI). Information on advising in primary health care was obtained asking whether GP advised overweight patients to change dietary habits and to increase physical activity. The odds of receiving advice on diet and physical activity were calculated using multiple logistic regression analyses according to a range of sociodemographic variables, perceived health, number of visits to GPs and body weight status. RESULTS: Almost a half of respondents were overweight or obese. Only one fourth of respondents reported that they were advised to change diet. The proportion of persons who received advice on physical activity was even lower. The odds of receiving advice increased with age. A strong association was found between perceived health and receiving advice. The likelihood of receiving advice was related to BMI. GPs were more likely to give advice when BMI was high. More than a half of obese respondents (63.3%) reported that they had tried to lose weight. The association between receiving advice and self-reported attempt to lose weight was found. CONCLUSION: The low rate of dietary and physical activity advice reported by overweight patients implies that more lifestyle counselling should be provided in primary health care. There is an obvious need for improved training and education of GPs in counselling of overweight patients focusing on methods of giving dietary and physical activity advice. PMID- 16478533 TI - Ethanol inhibits L1 cell adhesion molecule activation of mitogen-activated protein kinases. AB - Inhibition of the functions of L1 cell adhesion molecule (L1) by ethanol has been implicated in the pathogenesis of the neurodevelopmental aspects of the fetal alcohol syndrome (FAS). Ethanol at pharmacological concentrations has been shown to inhibit L1-mediated neurite outgrowth of rat post-natal day 6 cerebellar granule cells (CGN). Extracellular signal-related kinases (ERK) 1/2 activation occurs following L1 clustering. Reduction in phosphoERK1/2 by inhibition of mitogen-activated protein kinase kinase (MEK) reduces neurite outgrowth of cerebellar neurons. Here, we examine the effects of ethanol on L1 activation of ERK1/2, and whether this activation occurs via activation of fibroblast growth factor receptor 1 (FGFR1). Ethanol at 25 mm markedly inhibited ERK1/2 activation by both clustering L1 with cross-linked monoclonal antibodies, or by L1-Fc chimeric proteins. Clustering L1 with subsequent ERK1/2 activation did not result in tyrosine phosphorylation of the FGFR1. In addition, inhibition of FGFR1 tyrosine kinase blocked basic fibroblast growth factor (bFGF) activation of ERK1/2, but did not affect activation of ERK1/2 by clustered L1. We conclude that ethanol disrupts the signaling pathway between L1 clustering and ERK1/2 activation, and that this occurs independently of the FGFR1 pathway in cerebellar granule cells. PMID- 16478536 TI - CoryneRegNet: an ontology-based data warehouse of corynebacterial transcription factors and regulatory networks. AB - BACKGROUND: The application of DNA microarray technology in post-genomic analysis of bacterial genome sequences has allowed the generation of huge amounts of data related to regulatory networks. This data along with literature-derived knowledge on regulation of gene expression has opened the way for genome-wide reconstruction of transcriptional regulatory networks. These large-scale reconstructions can be converted into in silico models of bacterial cells that allow a systematic analysis of network behavior in response to changing environmental conditions. DESCRIPTION: CoryneRegNet was designed to facilitate the genome-wide reconstruction of transcriptional regulatory networks of corynebacteria relevant in biotechnology and human medicine. During the import and integration process of data derived from experimental studies or literature knowledge CoryneRegNet generates links to genome annotations, to identified transcription factors and to the corresponding cis-regulatory elements. CoryneRegNet is based on a multi-layered, hierarchical and modular concept of transcriptional regulation and was implemented by using the relational database management system MySQL and an ontology-based data structure. Reconstructed regulatory networks can be visualized by using the yFiles JAVA graph library. As an application example of CoryneRegNet, we have reconstructed the global transcriptional regulation of a cellular module involved in SOS and stress response of corynebacteria. CONCLUSION: CoryneRegNet is an ontology-based data warehouse that allows a pertinent data management of regulatory interactions along with the genome-scale reconstruction of transcriptional regulatory networks. These models can further be combined with metabolic networks to build integrated models of cellular function including both metabolism and its transcriptional regulation. PMID- 16478537 TI - Nosocomial bloodstream infections caused by Klebsiella pneumoniae: impact of extended-spectrum beta-lactamase (ESBL) production on clinical outcome in a hospital with high ESBL prevalence. AB - BACKGROUND: The frequency of ESBL producing Klebsiella pneumoniae bloodstream infections (BSI) is high in Brazilian hospitals, however little is known regarding what role, if any, resistance plays in the expected outcome in hospitals with a high prevalence of these pathogens. METHODS: From 1996 to 2001, hospital acquired K. pneumoniae BSI were evaluated retrospectively. Each patient was included only once at the time of BSI. ESBL producing strains were identified using the E-test method. The association of variables with the mortality related to bacteremia was included in a stepwise logistic regression model. RESULTS: One hundred and eight hospital acquired K. pneumoniae BSI met criteria for inclusion. Fifty two percent were due to ESBL producing strains. The overall in-hospital mortality was 40.8%. Variables independently predicting death by multivariate analysis were the following: mechanical ventilation (p = 0.001), number of comorbidities (p = 0.003), antimicrobials prescribed before bacteremia (p = 0.01) and fatal underlying disease (p = 0.025). CONCLUSION: Bacteremia due to ESBL producing K. pneumoniae strains was not an independent predictor for death in patients with BSI. An increased mortality in hospital-acquired BSI by K. pneumoniae was related to the requirement for mechanical ventilation, more than two comorbidities, the previous use of two or more antibiotics, and the presence of a rapidly fatal disease. PMID- 16478539 TI - Aluminum sulfate significantly reduces the skin test response to common allergens in sensitized patients. AB - BACKGROUND: Avoidance of allergens is still recommended as the first and best way to prevent allergic illnesses and their comorbid diseases. Despite a variety of attempts there has been very limited success in the area of environmental control of allergic disease. Our objective was to identify a non-invasive, non pharmacological method to reduce indoor allergen loads in atopic persons' homes and public environments. We employed a novel in vivo approach to examine the possibility of using aluminum sulfate to control environmental allergens. METHODS: Fifty skin test reactive patients were simultaneously skin tested with conventional test materials and the actions of the protein/glycoprotein modifier, aluminum sulfate. Common allergens, dog, cat, dust mite, Alternaria, and cockroach were used in the study. RESULTS: Skin test reactivity was significantly reduced by the modifier aluminum sulfate. Our studies demonstrate that the effects of histamine were not affected by the presence of aluminum sulfate. In fact, skin test reactivity was reduced independent of whether aluminum sulfate was present in the allergen test material or removed prior to testing, indicating that the allergens had in some way been inactivated. CONCLUSION: Aluminum sulfate was found to reduce the in vivo allergic reaction cascade induced by skin testing with common allergens. The exact mechanism is not clear but appears to involve the alteration of IgE-binding epitopes on the allergen. Our results indicate that it may be possible to diminish the allergenicity of an environment by application of the active agent aluminum sulfate, thus producing environmental control without complete removal of the allergen. PMID- 16478540 TI - Algebraic comparison of metabolic networks, phylogenetic inference, and metabolic innovation. AB - BACKGROUND: Comparison of metabolic networks is typically performed based on the organisms' enzyme contents. This approach disregards functional replacements as well as orthologies that are misannotated. Direct comparison of the structure of metabolic networks can circumvent these problems. RESULTS: Metabolic networks are naturally represented as directed hypergraphs in such a way that metabolites are nodes and enzyme-catalyzed reactions form (hyper)edges. The familiar operations from set algebra (union, intersection, and difference) form a natural basis for both the pairwise comparison of networks and identification of distinct metabolic features of a set of algorithms. We report here on an implementation of this approach and its application to the procaryotes. CONCLUSION: We demonstrate that metabolic networks contain valuable phylogenetic information by comparing phylogenies obtained from network comparisons with 16S RNA phylogenies. The algebraic approach to metabolic networks is suitable to study metabolic innovations in two sets of organisms, free living microbes and Pyrococci, as well as obligate intracellular pathogens. PMID- 16478538 TI - Phosphorylation-facilitated sumoylation of MEF2C negatively regulates its transcriptional activity. AB - BACKGROUND: Sumoylation has emerged as an important posttranslational regulatory mechanism for transcription factors and cofactors. Sumoylation of many transcription factors represses their transcriptional activities. The myocyte enhancer factor 2 (MEF2) family of transcription factors plays an important role in regulating gene expression during myogenesis and has been recently shown to be sumoylated. RESULTS: Consistent with earlier reports, we show that sumoylation of MEF2C at K391 inhibits its transcriptional activity. Sumoylation of MEF2C does not block its DNA-binding activity. A small C-terminal fragment of MEF2C containing K391, referred to as delta-N2-MEF2C, is efficiently sumoylated and, when targeted to DNA, represses transcription at neighbouring promoters. Because delta-N2-MEF2C lacks the binding site for class II histone deacetylases (HDACs), this result suggests that sumoylation of MEF2C may help to recruit transcriptional repressors other than these HDACs. Intriguingly, we show that phosphorylation of S396 in MEF2C, a residue in close proximity to the major sumoylation site (K391) and known to be phosphorylated in vivo, enhances sumoylation of delta- N2-MEF2C in vitro. The S396A mutation reduces sumoylation of MEF2C in vivo and enhances the transcription activity of MEF2C in reporter assays. CONCLUSION: We propose that phosphorylation of MEF2C at S396 facilitates its sumoylation at K391, which in turn recruits yet unidentified co-repressors to inhibit transcription. Our studies further suggest that sumoylation motifs containing a phosphorylated serine or an acidic residue at the +5 position might be more efficiently sumoylated. PMID- 16478541 TI - The Chronic Pain Coping Inventory: confirmatory factor analysis of the French version. AB - BACKGROUND: Coping strategies are among the psychosocial factors hypothesized to contribute to the development of chronic musculoskeletal disability. The Chronic Pain Coping Inventory (CPCI) was developed to assess eight behavioral coping strategies targeted in multidisciplinary pain treatment (Guarding, Resting, Asking for Assistance, Task Persistence, Relaxation, Exercise/Stretch, Coping Self-Statements and Seeking Social Support). The present study had two objectives. First, it aimed at measuring the internal consistency and the construct validity of the French version of the CPCI. Second, it aimed to verify if, as suggested by the CPCI authors, the scales of this instrument can be grouped according to the following coping families: Illness-focused coping and Wellness-focused coping. METHOD: The CPCI was translated into French with the forward and backward translation procedure. To evaluate internal consistency, Cronbach's alphas were computed. Construct validity of the inventory was estimated through confirmatory factor analysis (CFA) in two samples: a group of 439 Quebecois workers on sick leave in the sub-acute stage of low back pain (less than 84 days after the work accident) and a group of 388 French chronic pain patients seen in a pain clinic. A CFA was also performed to evaluate if the CPCI scales were grouped into two coping families (i.e. Wellness-focused and Illness focused coping). RESULTS: The French version of the CPCI had adequate internal consistency in both samples. The CFA confirmed the eight-scale structure of the CPCI. A series of second-order CFA confirmed the composition of the Illness focused family of coping (Guarding, Resting and Asking for Assistance). However, the composition of the Wellness-focused family of coping (Relaxation, Exercise/Stretch, Coping Self-Statements and Seeking Social Support) was different than the one proposed by the authors of the CPCI. Also, a positive correlation was observed between Illness and Wellness coping families. CONCLUSION: The present study indicates that the internal consistency and construct validity of the French version of the CPCI were adequate, but the grouping and labeling of the CPCI families of coping are debatable and deserve further analysis in the context of musculoskeletal and pain rehabilitation. PMID- 16478543 TI - Na(v) 1.8-null mice show stimulus-dependent deficits in spinal neuronal activity. AB - BACKGROUND: The voltage gated sodium channel Na(v) 1.8 has a highly restricted expression pattern to predominantly nociceptive peripheral sensory neurones. Behaviourally Na(v) 1.8-null mice show an increased acute pain threshold to noxious mechanical pressure and also deficits in inflammatory and visceral, but not neuropathic pain. Here we have made in vivo electrophysiology recordings of dorsal horn neurones in intact anaesthetised Na(v) 1.8-null mice, in response to a wide range of stimuli to further the understanding of the functional roles of Na(v) 1.8 in pain transmission from the periphery to the spinal cord. RESULTS: Na(v) 1.8-null mice showed marked deficits in the coding by dorsal horn neurones to mechanical, but not thermal, -evoked responses over the non-noxious and noxious range compared to littermate controls. Additionally, responses evoked to other stimulus modalities were also significantly reduced in Na(v) 1.8-null mice where the reduction observed to pinch > brush. The occurrence of ongoing spontaneous neuronal activity was significantly less in mice lacking Na(v) 1.8 compared to control. No difference was observed between groups in the evoked activity to electrical activity of the peripheral receptive field. CONCLUSION: This study demonstrates that deletion of the sodium channel Na(v) 1.8 results in stimulus-dependent deficits in the dorsal horn neuronal coding to mechanical, but not thermal stimuli applied to the neuronal peripheral receptive field. This implies that Na(v) 1.8 is either responsible for, or associated with proteins involved in mechanosensation. PMID- 16478544 TI - Cytokine responses of bovine macrophages to diverse clinical Mycobacterium avium subspecies paratuberculosis strains. AB - BACKGROUND: Mycobacterium avium subsp. paratuberculosis (MAP), the causative agent of Johne's disease (JD) persistently infects and survives within the host macrophages. While it is established that substantial genotypic variation exists among MAP, evidence for the correlates that associate specific MAP genotypes with clinical or sub-clinical disease phenotypes is presently unknown. Thus we studied strain differences in intracellular MAP survival and host responses in a bovine monocyte derived macrophage (MDM) system. RESULTS: Intracellular survival studies showed that a bovine MAP isolate (B1018) and a human MAP isolate (Hu6) persisted in relatively higher numbers when compared with a sheep MAP isolate (S7565) at 24 hr, 48-hr and 96-hr post infection (PI). MDMs stimulated with B1018 up-regulated IL-10 at the transcript level and down-regulated TNFalpha at the protein and transcript levels compared with stimulations by the S7565 and Hu6. MDMs infected with Hu6 showed a down regulatory pattern of IL-10 and TNFalpha compared to stimulations by S7565. Cells stimulated with B1018 and Hu6 had low levels of matrix metalloprotease-3 (MMP3) and high levels of tissue inhibitor of metalloprotease-1 (TIMP1) at 96-hr PI relative to MDMs stimulated by S7565. CONCLUSION: Taken together, results suggest that the bovine (B1018) and the human (Hu6) MAP isolates lead to anti-inflammatory and anti-invasive pathways in the macrophage environment whereas the sheep (S7565) MAP isolate induces a pro inflammatory pathway. Thus the infecting strain genotype may play a role in polarizing the host immune responses and dictate the clinicopathological outcomes in this economically important disease. PMID- 16478545 TI - Prediction of protein continuum secondary structure with probabilistic models based on NMR solved structures. AB - BACKGROUND: The structure of proteins may change as a result of the inherent flexibility of some protein regions. We develop and explore probabilistic machine learning methods for predicting a continuum secondary structure, i.e. assigning probabilities to the conformational states of a residue. We train our methods using data derived from high-quality NMR models. RESULTS: Several probabilistic models not only successfully estimate the continuum secondary structure, but also provide a categorical output on par with models directly trained on categorical data. Importantly, models trained on the continuum secondary structure are also better than their categorical counterparts at identifying the conformational state for structurally ambivalent residues. CONCLUSION: Cascaded probabilistic neural networks trained on the continuum secondary structure exhibit better accuracy in structurally ambivalent regions of proteins, while sustaining an overall classification accuracy on par with standard, categorical prediction methods. PMID- 16478546 TI - How much attention is needed towards men who sell sex to men for HIV prevention in India? AB - BACKGROUND: HIV prevention in India has mostly focussed on heterosexual transmission. Data on homosexual transmission are not readily available from India. We therefore assessed the probability of acquiring and transmitting HIV for men who sell sex to men and compared this with women who sell sex in India. METHODS: Sexual behaviour characteristics of 6661 men who have sex with men and 6648 women who sell sex were obtained in the Indian state of Andhra Pradesh through confidential interviews. These, along with estimates of HIV rates among them and risk of HIV transmission per unprotected sex act from other sources, were used to calculate their annual probability of acquiring and transmitting HIV. RESULTS: Of 6661 men who have sex with men in this sample, 1776 (26.7%) had sold sex to men. For every 1000 men who sell sex to men, annually 146 (95% confidence interval [CI] 116-179) would acquire HIV and HIV would be transmitted to 55 (95% CI 42-71) men who do not sell sex or women. These estimates were higher by 6.7 (95% CI 4.9-9.2) times for acquiring HIV and 2.5 (95% CI 2.0-3.2) times for transmitting HIV to sex partners outside their group, as compared with similar estimates for women who sell sex. In this sample, the average annual probability of acquiring HIV was higher among men who have sex with men but do not sell sex as compared with women who sell sex. CONCLUSION: These data indicate that men who sell sex to men are at much higher risk of acquiring and transmitting HIV than women who sell sex. Therefore, men who sell sex to men and their clients warrant substantial attention for comprehensive HIV prevention in India. PMID- 16478547 TI - In search of functional association from time-series microarray data based on the change trend and level of gene expression. AB - BACKGROUND: The increasing availability of time-series expression data opens up new possibilities to study functional linkages of genes. Present methods used to infer functional linkages between genes from expression data are mainly based on a point-to-point comparison. Change trends between consecutive time points in time-series data have been so far not well explored. RESULTS: In this work we present a new method based on extracting main features of the change trend and level of gene expression between consecutive time points. The method, termed as trend correlation (TC), includes two major steps: 1, calculating a maximal local alignment of change trend score by dynamic programming and a change trend correlation coefficient between the maximal matched change levels of each gene pair; 2, inferring relationships of gene pairs based on two statistical extraction procedures. The new method considers time shifts and inverted relationships in a similar way as the local clustering (LC) method but the latter is merely based on a point-to-point comparison. The TC method is demonstrated with data from yeast cell cycle and compared with the LC method and the widely used Pearson correlation coefficient (PCC) based clustering method. The biological significance of the gene pairs is examined with several large-scale yeast databases. Although the TC method predicts an overall lower number of gene pairs than the other two methods at a same p-value threshold, the additional number of gene pairs inferred by the TC method is considerable: e.g. 20.5% compared with the LC method and 49.6% with the PCC method for a p-value threshold of 2.7E-3. Moreover, the percentage of the inferred gene pairs consistent with databases by our method is generally higher than the LC method and similar to the PCC method. A significant number of the gene pairs only inferred by the TC method are process-identity or function-similarity pairs or have well-documented biological interactions, including 443 known protein interactions and some known cell cycle related regulatory interactions. It should be emphasized that the overlapping of gene pairs detected by the three methods is normally not very high, indicating a necessity of combining the different methods in search of functional association of genes from time-series data. For a p-value threshold of 1E-5 the percentage of process-identity and function-similarity gene pairs among the shared part of the three methods reaches 60.2% and 55.6% respectively, building a good basis for further experimental and functional study. Furthermore, the combined use of methods is important to infer more complete regulatory circuits and network as exemplified in this study. CONCLUSION: The TC method can significantly augment the current major methods to infer functional linkages and biological network and is well suitable for exploring temporal relationships of gene expression in time-series data. PMID- 16478548 TI - Molecular epidemiology and emergence of worldwide epidemic clones of Neisseria meningitidis in Taiwan. AB - BACKGROUND: Meningococcal disease is infrequently found in Taiwan, a country with 23 million people. Between 1996 and 2002, 17 to 81 clinical cases of the disease were reported annually. Reported cases dramatically increased in 2001-2002. Our record shows that only serogroup B and W135 meningococci have been isolated from patients with meningococcal disease until 2000. However, serogroup A, C and Y meningococci were detected for the first time in 2001 and continued to cause disease through 2002. Most of serogroup Y meningococcus infections localized in Central Taiwan in 2001, indicating that a small-scale outbreak of meningococcal disease had occurred. The occurrence of a meningococcal disease outbreak and the emergence of new meningococcal strains are of public health concern. METHODS: Neisseria meningitidis isolates from patients with meningococcal disease from 1996 to 2002 were collected and characterized by serogrouping, pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The genetic relatedness and clonal relationship between the isolates were analyzed by using the PFGE patterns and the allelic profiles of the sequence types (STs). RESULTS: Serogroups A, B, C, W135, Y, and non-serogroupable Neisseria meningitidis were, respectively, responsible for 2%, 50%, 2%, 35%, 9%, and 2% of 158 culture confirmed cases of meningococcal disease in 1996-2002. Among 100 N. meningitidis isolates available for PFGE and MLST analyses, 51 different PFGE patterns and 30 STs were identified with discriminatory indices of 0.95 and 0.87, respectively. Of the 30 STs, 21 were newly identified and of which 19 were found in serogroup B isolates. A total of 40 PFGE patterns were identified in 52 serogroup B isolates with the patterns distributed over several distinct clusters. In contrast, the isolates within each of the serogroups A, C, W135, and Y shared high levels of PFGE pattern similarity. Analysis of the allelic profile of the 30 STs suggested the serogroup B isolates be assigned into 5 clonally related groups/ clonal complexes and 7 unique clones. The ST-41/44 complex/Lineage 3, and the ST-3439 and ST-3200 groups represented 79% of the serogroup B meningococci. In contrast, isolates within serogroups A, serogroup W135 (and C), and serogroup Y, respectively, simply belonged to ST-7, ST-11, and ST-23 clones. CONCLUSION: Our data suggested that serogroup B isolates were derived from several distinct lineages, most of which could either be indigenous or were introduced into Taiwan a long time ago. The serogroup A, W135 (and C), and Y isolates, respectively, belonged to the ST-7, ST-11, and ST-23, and the represented clones that are currently the major circulating clones in the world and are introduced into Taiwan more recently. The emergence of serogroup A, C and Y strains contributed partly to the increase in cases of meningococcal disease in 2001-2002. PMID- 16478550 TI - Infant and maternal behavior moderate reactivity to novelty to predict anxious behavior at 2.5 years. AB - The degree to which infant regulatory behaviors, together with infant reactivity to novelty, predicted anxious behavior at 2.5 years, and the moderating effect of maternal behavior were tested. Sixty-four low-risk mothers and infants participated. Mothers rated infant negative reactivity and anxious behavior; infant and maternal behaviors were observed at 6 months postpartum. Based on results of hierarchical, multiple regressions, infant regulatory behaviors (i.e., attention control, withdrawal) moderated associations between reactivity to novelty and later anxious behavior, but predictions depended also on maternal behavior. High reactivity to novelty, in conjunction with withdrawal and with poor attention control, predicted anxious behavior only when mothers were less engaged or less sensitive, suggesting that maternal behavior alters developmental trajectories associated with infant temperament. PMID- 16478549 TI - Maternal depressive symptoms and child sleep: models of mutual influence over time. AB - The aim of the research was to test hypotheses concerning the associations between maternal depressive symptoms and child sleep using longitudinal data to examine possible predictive pathways. Data from the National Institute of Child Health and Human Development Study of Early Child Care with 1222 children studied from 1 to 36 months of age were analyzed to examine: effects on trajectories over time, and phase-specific effects over three defined age periods (6 to 15, 15 to 24, and 24 to 36 months). Child sleep was found to influence maternal depressive symptoms only for the 15- to 24-month age period, where, contrary to expectation, longer duration of child awakenings predicted decreased maternal depressive symptoms. Maternal depressive symptoms were found to predict an increased frequency of child awakenings across the 15- to 24-month age period only. In contrast, maternal depressive symptoms were found to significantly predict increased duration of child awakenings both for the 3-year trajectory and across the 15- to 24- and 24- to 36-month age periods. Additional research is needed to clarify the mechanisms by which maternal depressive symptoms predict increased duration of child awakenings. PMID- 16478551 TI - Developmental trajectories of externalizing and internalizing behaviors: factors underlying resilience in physically abused children. AB - Using a multisite community sample of 585 children, this study examined how protective and vulnerability factors alter trajectories of teacher-reported externalizing and internalizing behavior from kindergarten through Grade 8 for children who were and were not physically abused during the first 5 years of life. Early lifetime history of physical abuse (11.8% of sample) was determined through interviews with mothers during the prekindergarten period; mothers and children provided data on vulnerability and protective factors. Regardless of whether the child was abused, being African American; being male; having low early social competence, low early socioeconomic status (SES), and low adolescent SES; and experiencing adolescent harsh discipline, low monitoring, and low parental knowledge were related to higher levels of externalizing problems over time. Having low early social competence, low early SES, low adolescent SES, and low proactive parenting were related to higher levels of internalizing problems over time. Furthermore, resilience effects, defined as significant interaction effects, were found for unilateral parental decision making (lower levels are protective of externalizing outcomes for abused children), early stress (lower levels are protective of internalizing outcomes for abused children), adolescent stress (lower levels are protective of internalizing outcomes for abused children), and hostile attributions (higher levels are protective of internalizing outcomes for abused children). The findings provide a great deal of support for an additive or main effect perspective on vulnerability and protective factors and some support for an interactive perspective. It appears that some protective and vulnerability factors do not have stronger effects for physically abused children, but instead are equally beneficial or harmful to children regardless of their abuse status. PMID- 16478542 TI - The cancer stem cell: evidence for its origin as an injured autoreactive T cell. AB - This review explores similarities between lymphocytes and cancer cells, and proposes a new model for the genesis of human cancer. We suggest that the development of cancer requires infection(s) during which antigenic determinants from pathogens mimicking self-antigens are co-presented to the immune system, leading to breaking T cell tolerance. Some level of autoimmunity is normal and necessary for effective pathogen eradication. However, autoreactive T cells must be eliminated by apoptosis when the immune response is terminated. Apoptosis can be deficient in the event of a weakened immune system, the causes of which are multifactorial. Some autoreactive T cells suffer genomic damage in this process, but manage to survive. The resulting cancer stem cell still retains some functions of an inflammatory T cell, so it seeks out sites of inflammation inside the body. Due to its defective constitutive production of inflammatory cytokines and other growth factors, a stroma is built at the site of inflammation similar to the temporary stroma built during wound healing. The cancer cells grow inside this stroma, forming a tumor that provides their vascular supply and protects them from cellular immune response. As cancer stem cells have plasticity comparable to normal stem cells, interactions with surrounding normal tissues cause them to give rise to all the various types of cancers, resembling differentiated tissue types. Metastases form at an advanced stage of the disease, with the proliferation of sites of inflammation inside the body following a similar mechanism. Immunosuppressive cancer therapies inadvertently re-invigorate pathogenic microorganisms and parasitic infections common to cancer, leading to a vicious circle of infection, autoimmunity and malignancy that ultimately dooms cancer patients. Based on this new understanding, we recommend a systemic approach to the development of cancer therapies that supports rather than antagonizes the immune system. PMID- 16478553 TI - Multiple cognitive capabilities/deficits in children with an autism spectrum disorder: "weak" central coherence and its relationship to theory of mind and executive control. AB - This study examined the validity of "weak" central coherence (CC) in the context of multiple cognitive capabilities/deficits in autism. Children with an autism spectrum disorder (ASD) and matched typically developing children were administered tasks tapping visuospatial coherence, false-belief understanding and aspects of executive control. Significant group differences were found in all three cognitive domains. Evidence of local processing on coherence tasks was widespread in the ASD group, but difficulties in attributing false beliefs and in components of executive functioning were present in fewer of the children with ASD. This cognitive profile was generally similar for younger and older children with ASD. Furthermore, weak CC was unrelated to false-belief understanding, but aspects of coherence (related to integration) were associated with aspects of executive control. Few associations were found between cognitive variables and indices of autistic symptomatology. Implications for CC theory are discussed. PMID- 16478552 TI - The impact of foster care on development. AB - Foster care is a protective intervention designed to provide out of home placement to children living in at-risk home environments. This study employs prospective longitudinal data (N = 189) to investigate the effects of foster care on the development of child behavior and psychological functioning taking into account baseline adaptation prior to placement and socioeconomic status at the time of placement. Comparisons were made among three groups: children who experienced foster care, those who were maltreated but remained in the home, and children who had not experienced foster care or maltreatment despite their similarly at-risk demographic characteristics. In the current sample, children placed in out of home care exhibited significant behavior problems in comparison to children who received adequate care, and using the same pre- and postplacement measure of adaptation, foster care children showed elevated levels of behavior problems following release from care. Similarly, children placed into unfamiliar foster care showed higher levels of internalizing problems compared with children reared by maltreating caregivers, children in familiar care, and children who received adequate caregiving. Findings suggest that outcomes related to foster care may vary with type of care and beyond the effects associated with maltreatment history, baseline adaptation, and socioeconomic status. PMID- 16478554 TI - Meanings in motion and faces: developmental associations between the processing of intention from geometrical animations and gaze detection accuracy. AB - Aspects of face processing, on the one hand, and theory of mind (ToM) tasks, on the other hand, show specific impairment in autism. We aimed to discover whether a correlation between tasks tapping these abilities was evident in typically developing children at two developmental stages. One hundred fifty-four normal children (6-8 years and 16-18 years) and 13 high-IQ autistic children (11-17 years) were tested on a range of face-processing and IQ tasks, and a ToM test based on the attribution of intentional movement to abstract shapes in a cartoon. By midchildhood, the ability accurately and spontaneously to infer the locus of attention of a face with direct or averted gaze was specifically associated with the ability to describe geometrical animations using mental state terms. Other face-processing and animation descriptions failed to show the association. Autistic adolescents were impaired at both gaze processing and ToM descriptions, using these tests. Mentalizing and gaze perception accuracy are associated in typically developing children and adolescents. The findings are congruent with the possibility that common neural circuitry underlies, at least in part, processing implicated in these tasks. They are also congruent with the possibility that autism may lie at one end of a developmental continuum with respect to these skills, and to the factor(s) underpinning them. PMID- 16478555 TI - Peer victimization in early adolescence: association between physical and relational victimization and drug use, aggression, and delinquent behaviors among urban middle school students. AB - This study examined associations between two forms of peer victimization, physical and relational, and externalizing behaviors including drug use, aggression, and delinquent behaviors among a sample of 276 predominantly African American eighth graders attending middle school in an urban public school system. Regression analyses indicated that physical victimization was significantly related to cigarette and alcohol use but not to advanced alcohol and marijuana use; relational victimization contributed uniquely to all categories of drug use after controlling for physical victimization. Physical victimization was also significantly related to physical and relational aggression and delinquent behaviors, and relational victimization made a unique contribution in the concurrent prediction of these behaviors. Physical victimization was more strongly related to both categories of alcohol use, aggression, and to delinquent behaviors among boys than among girls. In contrast, relational victimization was more strongly related to physical aggression and marijuana use among girls than among boys, but more strongly related to relational aggression among boys than among girls. These findings provide information about the generalizability of prior research and have important implications for intervention efforts. PMID- 16478556 TI - The contribution of peers to monthly variation in adolescent depressed mood: a short-term longitudinal study with time-varying predictors. AB - This study examined peer predictors of variation and growth in depressed mood among high-risk adolescents, using child and parent reports of monthly symptoms. One hundred seventy-six parents and their 10- to 14-year-old children separately took part in a series of up to nine monthly interviews. Multilevel growth models examined both time-varying peer predictors of parent and child reports of the child's depressive symptoms, controlling for age, gender, and treatment status. Deviant peer affiliation significantly predicted elevated depressive symptoms in the monthly child-report of depressed mood, especially for younger adolescents. Children's level of delinquency was significantly related to parent-reported depressive symptoms, and to child-reported symptoms in older adolescents only. As expected, depressed mood was higher for girls and more prevalent among older adolescents. The results suggest that peer processes may be linked in time to the development of depression, especially among high-risk adolescents. PMID- 16478557 TI - Leaders and followers in adolescent close friendships: susceptibility to peer influence as a predictor of risky behavior, friendship instability, and depression. AB - Adolescents' susceptibility to peer influence was examined as a marker of difficulties in the general process of autonomy development that was likely to be related to deficits across multiple domains of psychosocial functioning. A laboratory-based assessment of susceptibility to peer influence in interactions with a close friend was developed and examined in relation to corollary reports obtained from adolescents, their mothers, and close peers at ages 13 and 14. As hypothesized, observed susceptibility to peer influence with a close friend predicted future responses to negative peer pressure, but it was also related to broader markers of problems in functioning, including decreases in popularity, and increasing levels of depressive symptoms, over time. Susceptibility to peer influence was also linked to higher concurrent levels of substance use, externalizing behavior, and sexual activity. Results are interpreted as reflecting the central role of establishing autonomy with peers in psychosocial development. PMID- 16478559 TI - Psychological profiles and adolescent adjustment: a person-centered approach. AB - The association between young adolescents' psychological profiles and their subsequent adjustment was examined in a sample of 606 adolescents (ages 12-13) drawn from the mother-child data set of the National Longitudinal Survey of Youth. Cluster analysis was used to identify distinct groups of youth based on self-regulation, proneness to risk, self-worth, and perceived academic competence. Five replicable clusters were identified corresponding to optimal, average, behavioral risk, low self-regulation, and emotional risk groups. These clusters were associated with distinct patterns of adjustment 4 years later. At ages 16-17, youth in the optimal group tended to report better academic performance, less problem behavior, and less depression than youth in the three risk groups; however, their functioning did not differ significantly from youth in the average group. The three risk groups differed in self-reported depression symptoms and academic performance but not in levels of problem behavior. Differences among the five groups persisted when demographic and contextual variables were controlled. These results support the existence of different groups of youth who follow distinct developmental trajectories and may experience different patterns of adjustment. PMID- 16478558 TI - Maternal and environmental factors influence the hypothalamic-pituitary-adrenal axis response to corticotropin-releasing hormone infusion in offspring of mothers with or without mood disorders. AB - Individuals with melancholic major depression exhibit basal hypercortisolism and an attenuated ACTH response to exogenous corticotropin-releasing hormone (CRH) infusion. Given the greater incidence of depression in children of depressed parents, we examined the ACTH and cortisol responses to ovine CRH (oCRH) infusion in 63 adolescent offspring of mothers with major depression, bipolar illness, or no psychiatric illness. Psychiatric and observational assessments of these families had been conducted over the course of 10 years preceding this study. We examined the children's responses to CRH in relation to maternal characteristics and family environment and found the following: (a) cortisol responses were negatively related to chronic family stress and (b) offspring of depressed mothers with an avoidant personality disorder showed an exaggerated ACTH response. In addition, adolescents in late puberty (Tanner 4 and 5) had lower ACTH and cortisol responses to oCRH infusion than those in early puberty. Further, offspring with early histories of mood problems, and those who developed major depressive disorder as young adults, did not exhibit basal hypercortisolism but did show an attenuated ACTH response to CRH. Our results add to the growing body of literature showing the influence of maternal characteristics and environmental factors on hypothalamic-pituitary-adrenal axis patterns in children. PMID- 16478560 TI - The mediating role of alienation in the development of maladjustment in youth exposed to community violence. AB - A study was conducted to determine predictors of high risk/delinquent behavior and psychoemotional maladjustment in 1,478 sixth and eighth graders who took part in a survey of high-risk and adaptive behaviors. The first goal was to determine the extent to which two distinct dimensions of alienation, normlessness, and isolation/self-estrangement mediated the emergence of maladjustment in youth exposed to community violence. The second goal was to distinguish between violence witnessing and violence victimization, and determine whether these subtypes of violence exposure may differentially affect the proposed mediational factors. The results indicated that normlessness partially mediated the relationship between violence exposure and high-risk/delinquent behavior and between violence exposure and psychoemotional maladjustment. Isolation/self estrangement partially mediated the relation between violence exposure and psychoemotional maladjustment. Although witnessing of, and victimization by, violence were both significant predictors of normlessness, only violent victimization was significantly linked to isolation/self-estrangement. These results elucidate the mechanisms by which community violence exposure may exert negative influences on youth and suggest that there are distinct patterns of risk associated with different forms of exposure. PMID- 16478561 TI - Dissociation following traumatic medical treatment procedures in childhood: a longitudinal follow-up. AB - Chronic illnesses often involve repeated hospitalization and invasive treatment procedures that can have a traumatic impact on child development. To explore possible consequences of treatment procedures, three groups of patients with congenital anomalies were examined longitudinally. At first admission, adolescents (ages 10-20, mean 15) with anorectal anomalies (n = 14), adolescents with Hirschsprung disease (n = 14), and hospitalized controls (n = 14) were assessed for treatment procedures, somatic function, mental health, and dissociative experiences. The assessment included the Adolescent Dissociative Experiences Scale (A-DES). At 10-year follow-up, the patients completed the Dissociative Experiences Scale (DES) and the Somatoform Dissociative Questionnaire (SDQ-20). Anal dilatation, an invasive medical treatment procedure performed daily by the parents the first 4 years, was correlated with the frequency and severity of persisting dissociative symptomatology. The procedure was the only significant predictor of A-DES and SDQ-20 scores, and one of two significant predictors of DES scores. This "experiment of nature" permitted a specific and unique opportunity to examine the impact of early traumatic exposure on child development in the absence of parental malevolence, and on later dissociative outcome in adolescence and adulthood. The findings might be valuable theoretically to our understanding of the development of psychopathology, and may lend itself for comparison with data on sexually abused children. PMID- 16478562 TI - Trajectories of depressive symptoms and stressful life events among male and female adolescents in divorced and nondivorced families. AB - This 11-year longitudinal study models the trajectories of depressive symptoms among approximately 550 females and males raised in divorced and nondivorced families in the rural Midwest. Using multilevel analyses, we demonstrate that, first, depressive symptoms changed according to a curvilinear pattern, especially for females; they increased during early to midadolescence and then declined in late adolescence to young adulthood. Second, compared with males, females experienced a greater number of depressive symptoms in adolescence and early adulthood. Third, children who experienced parental divorce by age 15 manifested a sharper increase in depressive symptoms compared to those from nondivorced families. Fourth, stressful life events children experienced shortly after parental divorce mediated the effect of parental divorce on depressive symptoms. Fifth and finally, time-varying stressful life events, particularly those related to relationship and personal loss, were significantly associated with the trajectories of depressive symptoms. PMID- 16478563 TI - Gender differences in the transition to early parenthood. AB - Data gathered over the course of a 25-year longitudinal study of 1,055 young people was used to examine gender differences in the onset of early parenthood and the developmental processes that place males and females at risk of becoming a young parent. Results revealed clear gender differences in the timing of early parenthood, with females being twice as likely as males to become a parent between the ages of 16 and 25 years. In contrast, the risk factors and life course processes that placed males and females at risk of an early transition to parenthood were very similar. Two exceptions were a gender-specific effect for maternal age and exposure to parental change, suggesting that having been raised by a younger mother and having experienced parental changes in your family of origin increased risks of early parenthood for females but not males. These findings contribute to our understanding of the effects of gender on life course development. PMID- 16478564 TI - Early parental loss and salivary cortisol in young adulthood: the moderating role of family environment. AB - Early family life adversity has been linked with negative physical and psychological health consequences in adulthood, possibly due to alterations in neuroendocrine activity. Young adults from families characterized by parental loss (N = 45) and control participants (N = 43) completed self-report measures of prior abuse and family conflict, and performed a stressful speech task designed to elicit neuroendocrine responses. Higher reported abuse and conflict were associated with increased cortisol for the loss group, but were unrelated to cortisol in the control group. Results indicate alterations in neuroendocrine functioning associated with early parental loss, which are moderated by the quality of the family environment. PMID- 16478565 TI - Immunological and pathological responses in BALB/c mice induced by genetic administration of Tc 13 Tul antigen of Trypanosoma cruzi. AB - Tc13 is a trans-sialidase family protein of Trypanosoma cruzi, the aetiological agent of Chagas' disease. Recently, in vitro studies had suggested that Tc13 might participate in the pathogenesis of the disease. In order to study the role of Tc13 antigens in an in vivo model, we administered plasmid DNA encoding a Tc13 antigen from the Tulahuen strain (Tc13 Tul) to BALB/c mice and evaluated the immunological and pathological manifestations as well as the capacity of this antigen to confer protection against T. cruzi infection. Tc13 Tul immunization did not elicit a detectable humoral immune response but induced specific memory T cells with no capacity to produce IFN-gamma. Five months after DNA-immunization with Tc13 Tul, signs of hepatotoxicity and reactive changes in the heart, liver and spleen were observed in 40-80% of mice. When Tc13 Tul DNA-immunized animals were challenged with trypomastigotes, a significant decrease in parasitaemia in early and late acute phase was observed without modification in the survival rate. Surprisingly, Tc13 Tul-immunized mice chronically infected with T. cruzi showed a decrease in the severity of heart damage. We conclude that, in BALB/c mice, genetic immunization with Tc13 Tul mainly induces immune responses associated with pathology. PMID- 16478566 TI - Two-photon fluorescence excitation and related techniques in biological microscopy. AB - This review is concerned with two-photon excited fluorescence microscopy (2PE) and related techniques, which are probably the most important advance in optical microscopy of biological specimens since the introduction of confocal imaging. The advent of 2PE on the scene allowed the design and performance of many unimaginable biological studies from the single cell to the tissue level, and even to whole animals, at a resolution ranging from the classical hundreds of nanometres to the single molecule size. Moreover, 2PE enabled long-term imaging of in vivo biological specimens, image generation from deeper tissue depth, and higher signal-to-noise images compared to wide-field and confocal schemes. However, due to the fact that up to this time 2PE can only be considered to be in its infancy, the advantages over other techniques are still being evaluated. Here, after a brief historical introduction, we focus on the basic principles of 2PE including fluorescence correlation spectroscopy. The major advantages and drawbacks of 2PE-based experimental approaches are discussed and compared to the conventional single-photon excitation cases. In particular we deal with the fluorescence brightness of most used dyes and proteins under 2PE conditions, on the optical consequences of 2PE, and the saturation effects in 2PE that mostly limit the fluorescence output. A complete section is devoted to the discussion of 2PE of fluorescent probes. We then offer a description of the central experimental issues, namely: choice of microscope objectives, two-photon excitable dyes and fluorescent proteins, choice of laser sources, and effect of the optics on 2PE sensitivity. An inevitably partial, but vast, overview of the applications and a large and up-to-date bibliography terminate the review. As a conclusive comment, we believe that 2PE and related techniques can be considered as a mainstay of the modern biophysical research milieu and a bright perspective in optical microscopy. PMID- 16478567 TI - White matter and subcortical gray matter lesion volume changes and late-life depression outcome: a 4-year magnetic resonance imaging study. AB - BACKGROUND: Cross-sectional studies have shown that late-onset depression is associated with larger deep white matter lesions (WMLs) and subcortical gray matter lesions (GMLs). In a longitudinal analysis, we examined changes in deep WMLs and subcortical GMLs in older depressed and nondepressed subjects over a 4 year period. METHODS: Brain magnetic resonance imaging (MRI) scans were obtained on 164 depressed and 126 healthy subjects aged 60 years or older at baseline, and 2 and 4 years after recruitment. WMLs and GMLs were measured using a semiautomated technique. We used repeated-measures analysis of covariance to determine cross-sectional lesion volume differences, whether lesion volume changes differed between patients and controls, and the effect of lesion volume changes on outcome in late-life depression. RESULTS: Mean volumes of lesions for the depressive group were 6.51, 8.18 and 7.75 cm2 for WMLs and 0.23, 0.30 and 0.34 cm2 for GMLs at baseline, 2-year and 4-year follow-up, respectively. Mean volumes of lesions for the control group were 4.83, 6.22 and 6.45 cm2 for WMLs and 0.17, 0.25 and 0.23 cm2 for GMLs at baseline, 2-year and 4-year follow-up, respectively. Cross-sectional between-group mean lesion volumes were significantly different for each measure. However, the pattern of WML and GML volume changes over time was not significantly different between groups. Treatment outcome was associated with changes in total and white matter lesion volume over time. CONCLUSIONS: Lesion volume progression is associated with aging and the pathological condition of late-life depression. The mechanisms that produce these progressive lesion changes remain unclear. Treatments aimed at arresting lesion progression may play a role in the management of late-life depression. PMID- 16478568 TI - Successful aging in Shanghai, China: definition, distribution and related factors. AB - OBJECTIVE: There are few studies of successful aging in China. This study was designed to investigate the distribution, and related factors, of successful aging in an elderly Chinese population. METHODS: A cross-sectional, community dwelling elderly population was surveyed in Shanghai, China. We defined successful aging based on a multi-dimensional model. Correlates of successful aging were explored through the Shanghai Successful Aging Project Questionnaire, which includes sociodemographic questions, and a battery of standardized instruments, including the Chinese version of the Mini-mental State Examination, activities of daily living, and the Life Satisfaction Index A (LSIA). RESULTS: The rate of successful aging was 46.2% [95% confidence interval (CI) 43.6-48.7] among people aged 65 or above, and the rate for males was higher than that for females. The rate was much lower for those aged 85 years or over (9.4%). Logistic regression analysis suggested that female gender and older age were unfavorable factors for successful aging. A higher score on the LSIA, more leisure activities and being currently married related to successful aging. CONCLUSION: The rate of successful aging in Shanghai, China is similar to that found in studies from western countries. There are some potentially modifiable factors that may relate to successful aging. PMID- 16478570 TI - Donepezil for negative signs in elderly patients with schizophrenia: an add-on, double-blind, crossover, placebo-controlled study. AB - OBJECTIVE: Cognitive impairment and negative signs are common in patients with schizophrenia. Up to 35% of elderly patients with schizophrenia fulfill the diagnostic criteria of dementia. Donepezil inhibits cholinesterase, thus enhancing cholinergic neurotransmission. We tested the efficacy of donepezil in elderly patients with chronic schizophrenia and severe cognitive impairment. METHOD: Following baseline assessment, patients were randomly assigned to receive either donepezil or placebo. The dose was 5 mg daily for the first week and 10 mg for an additional 11 weeks. The procedure was repeated using the crossover compound. The Positive and Negative Symptom Scale (PANSS), Clinical Global Impression Scale (CGI) and Alzheimer Disease Assessment Scale - Cognitive subscale (ADAS-Cog) were used to assess the severity of symptoms, cognitive status and intervention effects. RESULTS: Twenty subjects were enrolled (15 females, five males), mean age 70.2 years (SD 6.5) and mean duration of disease 38.5 years (SD 9.3). A modest treatment effect was found for both placebo and donepezil treatment periods. No crossover effect was found. No statistical differences were demonstrated between the two treatment groups (CGI p = 0.37, PANSS p = 0.71, ADAS-Cog p = 0.86). Two patients died during the study period due to unrelated causes and one patient discontinued participation due to increased agitation. CONCLUSION: Donepezil does not seem to improve negative signs and cognitive impairment in elderly patients with chronic schizophrenia. PMID- 16478571 TI - Differential item functioning related to education and age in the Italian version of the Mini-mental State Examination. AB - BACKGROUND: Differential item functioning (DIF) exists when test item responses by members of different demographic groups are statistically different when controlling for ability. DIF may indicate item bias. Our objective was to determine whether items from the Italian Mini-mental State Examination (MMSE) had DIF related to educational attainment, age, gender and occupation. We were also interested in exploring the significance of DIF in screening tests. METHODS: In a two-stage study from Granarolo, Italy, residents over age 61 (n = 495) were evaluated with the Italian MMSE. Those with MMSE scores of 28 or lower were further evaluated for dementia. MMSE results were coded in 10 item bundles. We used ordinal logistic regression to determine whether item bundles had DIF. RESULTS: Six of the 10 MMSE item bundles had DIF in educational attainment subgroups. Four of these six bundles also had DIF related to age. Items that required literacy were much harder for those with lower educational attainment. CONCLUSIONS: DIF related to education appeared at as few as 3 years of formal schooling. These findings suggest cautious interpretation of data from studies using the Italian MMSE in populations with heterogeneous educational backgrounds. DIF is especially troublesome for two-stage studies that use screening tests. PMID- 16478572 TI - Is the GDS-30 better than the GHQ-12 for screening depression in elderly people in the community? The Bambui Health Aging Study (BHAS). AB - BACKGROUND: Two-phase diagnostic surveys are popular in psychiatric epidemiology. The Geriatric Depression Scale (GDS) and the General Health Questionnaire (GHQ) are commonly used to screen in older and younger populations, respectively. METHOD: In Phase I, in this Brazilian population-based study, we screened 392 participants aged >or= 75 years. In Phase II, half of those scoring >or= 11 in the GDS and >or= 4 in the GHQ and 20% of others were selected for detailed evaluation with ICD-10 diagnoses assessed by the Schedules for Clinical Assessment in Neuropsychiatry (SCAN) (n = 126). RESULTS: Internal consistencies were good for all scales. At the optimal cut-off points GDS-30 (14/15) yielded 73% sensitivity and 65% specificity, while classical GHQ (4/5), Likert (15/16) and c-GHQ (5/6) yielded 66-75% sensitivity and 57-62% specificity. Receiver operating characteristic (ROC) curve analysis against SCAN indicated only moderate screening potential [area under the ROC curve (AUROC): GDS = 0.76; classical GHQ = 0.74; Likert = 0.76; c-GHQ = 0.73], with no statistically significant differences. All measures were biased by disability and self-reported health. CONCLUSIONS: In this study, neither screen was sufficiently predictive of SCAN ICD-10 diagnosis to recommend their use in two-phase surveys. Despite its theoretical advantages, the GDS-30 performed no better than the GHQ-12, and was biased in similar ways. PMID- 16478573 TI - Cloning and cheating. PMID- 16478575 TI - Weaknesses in the case against prepatterning in the mouse. AB - Serious weaknesses are exposed regarding recent claims to have demonstrated that there is no prepatterning of axes in the mouse. That the orientation of the axis of polarity of the blastocyst is dictated by the shape of the zona pellucida is contradicted by systematic observations on living conceptuses. Moreover, this 'mechanical constraint' hypothesis fails to account for key findings on which the case for prepatterning is based. PMID- 16478576 TI - Fatal flaws in the case for prepatterning in the mouse egg. AB - The presence or absence of predetermination and polarity in the mouse preimplantation embryo is still controversial. The question is if the mechanisms underlying early mammalian development is comparable to those operating in non mammalian 'model' organisms. In a recent article by Gardner in this journal, the author criticizes two of our recent publications. However, in order to resolve this controversy it is essential to read relevant reports carefully without bias and to provide data on which a particular claim is based. PMID- 16478577 TI - Oocyte and embryo donation 2006: reviewing two decades of innovation and controversy. AB - For over 21 years, oocyte and embryo donation have been used to treat infertility caused by a variety of conditions affecting the ovary. Many disorders, including premature ovarian failure, advanced reproductive age, unexplained recurrent implantation failure and inherited conditions, are amenable to gamete donation, with high pregnancy rates and good obstetrical outcomes observed in recipients. Protocols for the medical screening of recipients and donors, as well as infectious disease and genetic testing, have become relatively uniform and well accepted. Established guidelines allow synchronization of the menstrual cycles of both women to ensure that embryos are transferred to a receptive endometrium. The high demand for donor services has led to escalating costs and long waiting lists. American programmes bid against each other to secure the participation of young women often motivated as much by financial reward as altruism. In the United States, where the majority of oocyte donation is practised, more than 100,000 treatment cycles have occurred. However, to date no meaningful longitudinal studies detailing the long term effects of treatment on donors, recipients, children born, or families created have been published. Throughout its history, oocyte and embryo donation has proven to be both efficacious and clinically innovative, yet remains highly controversial. PMID- 16478578 TI - HMG versus rFSH for ovulation induction in developing countries: a cost effectiveness analysis based on the results of a recent meta-analysis. AB - Both cost and effectiveness should be considered conjointly to aid judgments about drug choice. Therefore, based on the results of a recent published meta analysis, a Markov model was developed to conduct a cost-effectiveness analysis for estimation of the cost of an ongoing pregnancy in IVF/intracytoplasmic sperm injection (ICSI) cycles. In addition, Monte Carlo micro-simulation was used to examine the potential impact of assumptions and other uncertainties represented in the model. The results of the study reveal that the estimated average cost of an ongoing pregnancy is 13,946 Egyptian pounds (EGP), and 18,721 EGP for a human menopausal gonadotrophin (HMG) and rFSH cycle respectively. On performing a sensitivity analysis on cycle costs, it was demonstrated that the rFSH price should be 0.61 EGP/IU to be as cost-effective as HMG at the price of 0.64 EGP/IU (i.e. around 60% reduction in its current price). The difference in cost between HMG and rFSH in over 100,000 cycles would result in an additional 4565 ongoing pregnancies if HMG was used. Therefore, HMG was clearly more cost-effective than rFSH. The decision to adopt a more expensive, cost-ineffective treatment could result in a lower number of cycles of IVF/ICSI treatment undertaken, especially in the case of most developing countries. PMID- 16478580 TI - Conception using vaginal administration of gonadotrophins in IVF: a case report. AB - A method has been developed of injecting gonadotrophins vaginally every 3-4 days, to reduce both the dose and frequency of injections. A couple with primary infertility for 5 years with a previous failed IVF cycle was recruited. A total dose of 1200 IU gonadotrophins was administered in three injections. Ten mature oocytes were recovered and six fertilized two-pronuclear stage zygotes resulted. The patient conceived after replacement of three good quality embryos. PMID- 16478583 TI - Use of ovarian reserve tests for the prediction of ongoing pregnancy in couples with unexplained or mild male infertility. AB - The chance of infertile patients conceiving is related to factors like female age and duration of infertility. This prospective observational study evaluated whether the results of ovarian reserve tests, including the novel marker serum anti-Mullerian hormone (AMH), were of additional value in predicting ongoing pregnancy. Two hundred and twenty-two patients diagnosed with unexplained infertility or mild male factor (total motile count>10x10(6)) on the basis of the infertility work-up were prospectively included. Antral follicle count, AMH, inhibin B, FSH and oestradiol concentrations were determined during the early follicular phase. Outcome measures were treatment-dependent and treatment independent ongoing pregnancy and time to ongoing pregnancy. There were 159 ongoing pregnancies, 52 of which occurred spontaneously. Pregnant patients were significantly younger than those who did not become pregnant (median age 32.4 versus 34.9 years, P<0.001) and FSH concentrations were higher in non-pregnant patients (median 6.8 versus 7.6 IU/l, P=0.04). Only age (hazard ratio 0.93, 95% CI 0.90-0.97) and whether or not the patient was undergoing treatment (hazard ratio 8.10, 95% CI 5.66-11.61) were significantly associated with time to ongoing pregnancy. Ovarian reserve tests, other than chronological age, seem of limited value in predicting (time to) ongoing pregnancy in patients with unexplained and mild male infertility. PMID- 16478582 TI - Menstrual effluent in endometriosis shows no difference in volume, VEGF-A, MMP2 and MMP9 or sFLT. AB - Since retrograde menstruation is considered a key event in the aetiology of endometriosis, this study sought to determine whether the menstrual effluent of women with this condition is different from that of those with a normal pelvis. As the amount of blood lost during menstruation is thought to be higher in this group, measured objective menstrual blood loss (MBL) was measured. In addition, factors enhancing both ectopic implantation of endometrium and its subsequent growth (by establishing a neo-vasculature) were chosen for study. Our hypothesis was that they are increased in the menstrual effluent of women with endometriosis. The study showed that at the time of menstruation, there is no difference in MBL or in the volume of menstrual effluent between women with endometriosis and those with a normal pelvis at laparoscopy. In addition, vascular endothelial growth factor-A (VEGF-A) message and protein, soluble truncated receptor sVEGF-R1 (sFLT), matrix metalloproteinase (MMP) 2 and MMP9 activities were also shown to be similar between the two groups. It is concluded that the enhanced expression of VEGF-A and MMP in the peritoneal fluid and ectopic lesions of endometriotic patients may be a secondary event, resulting from an innate difference in peritoneal and systemic factors rather than in the endometrium, causing an abnormal peritoneal response to menstrual debris and facilitating its ectopic implantation. PMID- 16478584 TI - Firm embryo transfer catheters for assisted reproduction: a systematic review and meta-analysis using direct and adjusted indirect comparisons. AB - A systematic review of published evidence on firm embryo transfer catheters is presented. Extensive searches were conducted for full-text manuscripts, abstracts, ongoing and unpublished trials. Direct and adjusted indirect comparisons were undertaken, where appropriate. Twenty-six randomized controlled trials comparing embryo transfer catheters were identified. Only two trials (314 transfers) compared different firm embryo catheters. Using direct comparison, both the Tom Cat and Tefcat catheters demonstrated statistically significant increased chances of clinical pregnancy compared with the Tight Difficult Transfer (TDT) catheter (P=0.007; OR=3.67, 95% CI=1.48-9.10 and P<0.0001; OR=4.71, 95% CI=2.34-9.48 respectively). The implantation rates were also higher with the Tom Cat and Tefcat catheters than the TDT catheter (P=0.005; OR=3.67, 95% CI=1.48-9.10 and P<0.00001; OR=4.29, 95% CI=2.45-7.50 respectively). Using adjusted indirect comparison, Tom Cat and Tefcat catheters were compared, and shown to have similar pregnancy and implantation rates (OR=0.99; 95% CI=-0.87 1.79 and OR=0.86; 95% CI=-0.77-1.35). In conclusion, both Tom Cat and Tefcat catheters give better outcomes than the TDT catheter, but are similar to each other. PMID- 16478585 TI - Endometrial polyps smaller than 1.5 cm do not affect ICSI outcome. AB - This study aimed to determine whether the presence of endometrial polyps discovered during ovarian stimulation affects the outcomes of intracytoplasmic sperm injection (ICSI) cycles. This retrospective descriptive study was conducted in a private assisted reproductive technology unit. Medical records of ICSI cycles performed between January 2003 and December 2004 were reviewed. Patients were divided into three groups: patients with endometrial polyps discovered during ovarian stimulation (group 1, n=15), patients who underwent hysteroscopic polyp resection prior to their ICSI cycle (group 2, n=40) and patients without polyps (group 3, n=956). Main outcome measures were clinical pregnancy rates and implantation rates. Age of the patients, age of the husbands, body mass index, total amount of gonadotrophins used, length of stimulation, peak oestradiol concentrations, peak endometrial thickness and number of embryos replaced were not significantly different between the groups, nor were the pregnancy and implantation rates. Only one patient (12.5%) from the first group experienced miscarriage within 12 weeks of pregnancy. In conclusion, endometrial polyps discovered during ovarian stimulation do not negatively affect pregnancy and implantation outcomes in ICSI cycles. PMID- 16478587 TI - Congenital partial atresia of the Fallopian tube. AB - Partial tubal atresia is a rare anomaly of unknown pathogenesis. It can occur unilaterally or bilaterally, mainly including the proximal isthmic portion or the proximal ampullary segment ('mid-portion'), and is especially associated with uterine anomalies. Any more extensive pelvic surgery, including preoperative diagnosis of associated urological anomalies, must be well planned in such patients. Salpingostomy and fimbrial approximation is a realistic option in patients with congenital ampullary atresia who wish to have a child. A MEDLINE search was performed and 18 patients found with partial atresia of different tubal portions. This report describes a further patient and reviews the available literature. PMID- 16478589 TI - Relationship between epidemiological features and aetiology of male infertility as diagnosed by a comprehensive infertility service provider. AB - This study examined the relationship between demographic features and aetiological causes of male infertility. Primary infertility was the presentation in 78% of patients. The incidence of varicocele was the highest (31%), whereas only 4.6% had vasectomy reversal and 7.4% of men were diagnosed with idiopathic infertility. Using the chi-squared test, there was no significant difference in the incidence of different causes of infertility among different ethnic groups (White, African-American, Asian, Hispanic, and other). Furthermore, there was no increased incidence of infertility aetiology with any particular occupation, race, religion, smoking or alcohol intake. In this study population, there was no association between the various risk factors (occupation, smoking, alcohol intake, and race) and the aetiologies of infertility. The proportion of patients diagnosed with idiopathic infertility was significantly less than reported in the literature. PMID- 16478590 TI - First pregnancy and life after preimplantation genetic diagnosis by polar body analysis for mucopolysaccharidosis type I. AB - Preimplantation genetic diagnosis (PGD) may help couples at risk to avoid pregnancies with known genetic diseases. In Germany, the only option to perform PGD is the analysis of polar bodies (PB). Mucopolysaccharidosis type I (MPS I) is an autosomal recessive lysosomal storage disorder. Q70X is one of the frequent diseases causing mutations of alpha-L-iduronidase (IDUA), leading to a severe phenotype with mental retardation and various somatic abnormalities, and making a request for PGD is understandable. Using five polymorphic DNA markers from the vicinity of IDUA, PGD on first PB was performed for a consanguineous couple, both heterozygotes of the Q70X mutation of IDUA. Sixteen first PB were obtained by laser assisted hatching of the zona pellucida. Genotyping led to the conclusion that 3/16 oocytes carried wild-type IDUA alleles. Only one of these oocytes showed pronucleus formation after intracytoplasmic sperm injection and was transferred on day 2 after oocyte retrieval. A singleton pregnancy was established. Prenatal diagnosis showed a fetus heterozygous for Q70X. For MPS I, PB analysis is a feasible way to perform PGD and it may be an acceptable alternative for couples with moral objections to embryo selection, or for countries in which genetic testing of the embryo is prohibited. PMID- 16478592 TI - Chromosome abnormalities and their relationship to morphology and development of human embryos. AB - This review covers the relationship between chromosome abnormalities, morphological abnormalities and embryonic development. The baseline of chromosome abnormalities in human embryos produced by assisted reproduction is higher than 50%, regardless of maternal age. While aneuploidy increases with maternal age, abnormalities arising post-meiotically, such as mosaicism, chaoticism, polyploidy and haploidy, have similar incidence in all age groups (about 33%). Post-meiotic abnormalities do increase with dysmorphism. The most common dysmorphisms found in cleavage-stage embryos are multinucleation, fragmentation and uneven cells, among others. All dysmorphisms are associated with an increase in post-meiotic chromosome abnormalities and a decreased implantation potential. Similarly, embryos developing slowly or with arrested development have higher incidence of post-meiotic abnormalities than normally developing ones. Chromosome studies in blastocysts indicate that mosaicism is the most common abnormality but that the load of abnormal cells decreases with increasing blastocyst quality. Regardless of blastocyst quality, more than 40% of mosaics are still chromosomally abnormal and will not implant or will spontaneously abort. Because aneuploidy is not related to cleavage stage dysmorphism and trisomies can reach blastocyst stage and beyond, morphological analysis is not enough to select against chromosome abnormalities, and thus preimplantation genetic diagnosis should be recommended in patients 35 and older. PMID- 16478591 TI - Exploiting LH in ovarian stimulation. AB - During intermediate-late phases of human folliculogenesis, LH plays a key role in promoting steroidogenesis and growth of the leading follicle. Ovarian stimulation for assisted reproduction techniques usually consists of administering exogenous FSH in a low LH environment. Although an impairment in LH-dependent paracrine activities would be expected, multiple follicular growth is efficiently achieved in almost all patients. Thus, there appears to be a discrepancy between classical folliculogenesis models and data from IVF. This study examines the 'interface' between basic endocrinological and clinical evidence, in an attempt to answer two questions: is there an LH therapeutic window, and if there is, how can this be exploited in the practice of assisted reproduction? It also reviews the evidence that specific subgroups of women may benefit from LH supplementation during ovarian stimulation. PMID- 16478594 TI - Rediscovering Boveri's centrosome in Ascaris (1888): its impact on human fertility and development. AB - We rediscover and review the brilliant work of Theodore Boveri, over a 100 years ago, on the centrosome of the round worm Ascaris and show how it impacts on our understanding of human fertilization and embryogenesis. Boveri was able to make fundamental predictions on the mechanics of fertilization and the dominant role of the sperm centrosome (Boveri's rule), which is now applicable to most animals. Using advanced digital imaging by light and electron microscopy, we explore centrosomal dynamics during Ascaris fertilization and the first cell cycle during cleavage. Twenty figures are presented in this visual publication. Humans follow Boveri's rule, as do most mammals excluding some rodents, and there is a remarkable similarity of the events of fertilization and cleavage in Ascaris and humans, the latter of which has been documented since 1991. The role of the sperm centrosome (centriole) in egg activation, polarity, embryogenesis, infertility and cancer is discussed. An attempt is made to portray the images Boveri may have visualized, in his painstaking drawings presented in his thesis in 1888. We now know the origins of the centrosome in human somatic cells--predominantly from the sperm cell. The impact of Boveri's work on human development is highlighted in this age of assisted reproductive technology. PMID- 16478596 TI - Dry storage of sperm: applications in primates and domestic animals. AB - Cryopreservation of spermatozoa, oocytes and embryos, as well as somatic cells or cell lines for cloning from cells, are all options for the long-term storage of unique genotypes and endangered species. Spermatozoal cryopreservation and storage currently require liquid nitrogen or ultralow refrigeration-based methods for long- or short-term storage, which requires routine maintenance and extensive space requirements. The preservation of stem cells also has strict requirements for long-term storage to maintain genetic integrity. Dessicated (lyopreserved) sperm and stem cells will provide an unprecedented type of long-term storage without the need for expensive and burdensome cryogenic conditions. Experiments were conducted to determine an effective intracellular concentration of the lyoprotectant trehalose. High-pressure liquid chromatography studies revealed that trehalose can be incorporated into mature sperm cells as well as spermatogonial stem cells from rhesus monkeys. In addition, using fourier transform infrared spectroscopy, we determined that thermotropic phase transitions for fresh ejaculates from rhesus monkey and stallion sperm occurred at 10-15, 33-37 and 55-59 degrees C. Preliminary studies in our laboratory have indicated that spermatogonial stem cells can be dried to <3 g g(-1) water and maintain viability following rehydration. Studies in our laboratory have provided preliminary results suggesting that the desiccated storage of sperm and spermatogonial stem cells may be a viable alternative to conventional cryopreservation. PMID- 16478597 TI - Spermatogonia: origin, physiology and prospects for conservation and manipulation of the male germ line. AB - In recent years, the scientific community has become increasingly interested in spermatogonia. Methodological breakthroughs, such as germ cell transplantation and spermatogonial culture combined with novel germ line transfection strategies, have provided interesting new opportunities for studying the physiology of spermatogonial stem cells and their interaction with the stem cell niche. Furthermore, intense research into pluripotent and adult stem cells has generated new insight into the differentiation pathway of germ line stem cells and has opened new perspectives for stem cell technologies. The present review briefly introduces the physiology of spermatogonial stem cells and discusses future directions of basic research and practical approaches applicable to livestock maintenance and animal reproduction. PMID- 16478598 TI - Male germ cell transplantation in livestock. AB - Male germ cell transplantation is a powerful approach to study the control of spermatogenesis with the ultimate goal to enhance or suppress male fertility. In livestock animals, applications can be expanded to provide an alternative method of transgenesis and an alternative means of artificial insemination (AI). The transplantation technique uses testis stem cells, harvested from the donor animal. These donor stem cells are injected into seminiferous tubules, migrate from the lumen to relocate to the basement membrane and, amazingly, they can retain the capability to produce donor sperm in their new host. Adaptation of the mouse technique for livestock is progressing, with gradual gains in efficiency. Germ cell transfer in goats has produced offspring, but not yet in cattle and pigs. In goats and pigs, the applications of germ cell transplantation are mainly in facilitating transgenic animal production. In cattle, successful male germ cell transfer could create an alternative to AI in areas where it is impractical. Large-scale culture of testis stem cells would enhance the use of elite bulls by providing a renewable source of stem cells for transfer. Although still in a developmental state, germ cell transplantation is an emerging technology with the potential to create new opportunities in livestock production. PMID- 16478600 TI - Moving to the beat: a review of mammalian sperm motility regulation. AB - Because it is generally accepted that a high percentage of poorly motile or immotile sperm will adversely affect male fertility, analysis of sperm motility is a central part of the evaluation of male fertility. In spite of its importance to fertility, poor sperm motility remains only a description of a pathology whose underlying cause is typically poorly understood. The present review is designed to bring the clinician up to date with the most current understanding of the mechanisms that regulate sperm motility and to raise questions about how aberrations in these mechanisms could be the underlying causes of this pathology. PMID- 16478599 TI - Sperm-mediated gene transfer. AB - Since 1989, a new method for the production of transgenic animals has been available, namely sperm-mediated gene transfer (SMGT), based on the intrinsic ability of sperm cells to bind and internalise exogenous DNA molecules and to transfer them into the oocyte at fertilisation. We first described the SMGT procedure in a small animal model, with high efficiency reported in the mouse. In addition, we successfully adapted and optimised the technique for use in large animals; it was, in fact, highly efficient in the generation of human decay accelerating factor transgenic pig lines, as well as multigene transgenic pigs in which three different reporter genes, namely enhanced green fluorescent protein, enhanced blue fluorescent protein and red fluorescent protein, were introduced. The major benefits of the SMGT technique were found to be its high efficiency, low cost and ease of use compared with other methods. Furthermore, SMGT does not require embryo handling or expensive equipment. Sperm-mediated gene transfer could also be used to generate multigene transgenic pigs that would be of benefit as large animal models for medical research, for agricultural and pharmaceutical applications and, in particular, for xenotransplantation, which requires extensive genetic manipulation of donor pigs to make them suitable for grafting to humans. PMID- 16478601 TI - Intracytoplasmic sperm injection in the bovine induces abnormal [Ca2+]i responses and oocyte activation. AB - Fertilisation by intracytoplasmic sperm injection (ICSI), a technique that bypasses the membrane fusion of the gametes, has been widely used to produce offspring in humans and mice. Success with this technique has lent support to the hypothesis that in mammalian fertilisation, a factor from the sperm, the so called sperm factor, is responsible for oocyte activation and that the fusion process is not involved in the generation of the hallmark [Ca2+]i signalling seen following fertilisation. However, the success of ICSI has largely eluded large domestic species, such as the bovine, porcine and equine, casting doubt on the current model of oocyte activation at fertilisation in these species. Using Ca2+ imagery and a series of treatments to manipulate the chemical structure of the sperm, we have investigated the early events of oocyte activation in response to ICSI in the bovine. Our results demonstrate, for the first time, that following ICSI, the majority of bovine oocytes are unable to mount [Ca2+]i oscillations, although, in few cases, the initiation of [Ca2+]i oscillations can occur in a manner indistinguishable from in vitro fertilisation. We also show that bull sperm possess a full complement of sperm factor. However, either the release and/or activation of the sperm factor are compromised after ICSI, leading to the delivery of a defective Ca2+ stimulus, which results in premature termination of embryo development. PMID- 16478602 TI - Mammalian membrane block to polyspermy: new insights into how mammalian eggs prevent fertilisation by multiple sperm. AB - To inhibit fertilisation by more than one sperm (a condition known as polyspermy), eggs have developed preventative mechanisms known as blocks to polyspermy. The block at the level of the egg extracellular coat (the zona pellucida in mammals, the vitelline envelope in non-mammals) has been well characterised in many different animal species and the block at the level of the egg plasma membrane is understood in some non-mammalian species. However, virtually nothing is known about the membrane block to polyspermy in mammalian eggs, despite data dating back 50-90 years that provide evidence for its existence. In the present review, we will discuss the background on blocks to polyspermy used by animal eggs and then focus on the membrane block to polyspermy in mammalian eggs. This will include a summary of classical studies that provide evidence for this block in mammalian eggs, assays used to study the mammalian membrane block and what has been elucidated from recent experimental studies about the cellular signalling events that lead to membrane block establishment and the mechanism of how the membrane block may prevent additional fertilisation. PMID- 16478603 TI - Gamete imprinting: setting epigenetic patterns for the next generation. AB - The acquisition of genomic DNA methylation patterns, including those important for development, begins in the germ line. In particular, imprinted genes are differentially marked in the developing male and female germ cells to ensure parent-of-origin-specific expression in the offspring. Abnormalities in imprints are associated with perturbations in growth, placental function, neurobehavioural processes and carcinogenesis. Based, for the most part, on data from the well characterised mouse model, the present review will describe recent studies on the timing and mechanisms underlying the acquisition and maintenance of DNA methylation patterns in gametes and early embryos, as well as the consequences of altering these patterns. PMID- 16478604 TI - Placental function in development and disease. AB - The placenta is an organ that clinicians and embryologists would all agree is important for pregnancy success. Unfortunately, however, they too often ignore it when they are exploring causes for embryonic, fetal and perinatal complications. The core function of the placenta is to mediate the transport of nutrients between the maternal and fetal circulation, but it also has critical endocrine functions that alter different maternal physiological systems in order to sustain pregnancy. Both its development and ongoing functions can be dynamically regulated by environmental factors, including nutrient status and tissue oxygenation. In recent years, mainstream attention has begun to shift onto the placenta and it is now becoming clear that placental pathology is associated with several complications in human and animal pregnancies, including embryonic lethality, fetal growth restriction, pre-eclampsia and the high rates of fetal deaths observed after nuclear transfer (cloning). PMID- 16478606 TI - Equine cloning: applications and outcomes. AB - Cloning is one of several new assisted reproductive techniques being developed for clinical use in the equine industry. Potential uses of equine cloning include: (1) the preservation of genetics from individual animals that would otherwise not be able to reproduce, such as geldings; (2) the preservation of genetic material of endangered and/or exotic species, such as the Mongolian wild horse (Przewalski's horse); and (3) because of the companion animal role that horses fill for some individuals, it is likely that some horse owners will have individual animals cloned for emotional fulfillment. Although equine cloning has been successful, like other species, it remains a very inefficient process (<3% success). In most species, the inefficiency of cloning results from a high incidence of embryonic, fetal and/or placental developmental abnormalities that contribute to extremely high rates of embryonic loss, abortion and stillbirths throughout gestation and compromised neonatal health after birth. The present review describes some of the ultrasonographic, endocrinological and histopathological characteristics of successful (produced viable offspring) and unsuccessful (resulted in pregnancy failure) cloned equine (mule and horse) pregnancies we have produced. A total of 21 cloned mule pregnancies were established using fetal fibroblast cells, whereas a total of seven cloned horse pregnancies were established using adult cumulus cells. Three of the cloned mule conceptuses were carried to term, resulting in the birth of three healthy clones. This information adds to an accumulating body of knowledge about the outcome of cloned equine pregnancies, which will help to establish when, and perhaps why, many cloned equine pregnancies fail. PMID- 16478608 TI - [Benign anorectal pathology]. PMID- 16478605 TI - Applications of emerging technologies to the study and conservation of threatened and endangered species. AB - Sustaining viable populations of all wildlife species requires the maintenance of habitat, as well as an understanding of the behaviour and physiology of individual species. Despite substantial efforts, there are thousands of species threatened by extinction, often because of complex factors related to politics, social and environmental conditions and economic needs. When species become critically endangered, ex situ recovery programmes that include reproductive scientists are the usual first line of defence. Despite the potential of reproductive technologies for rapidly increasing numbers in such small populations, there are few examples of success. This is not the result of a failure on the part of the technologies per se, but rather is due to a lack of knowledge about the fundamental biology of the species in question, information essential for allowing reproductive technologies to be effective in the production of offspring. In addition, modern conservation concepts correctly emphasise the importance of maintaining heterozygosity to sustain genetic vigour, thereby limiting the practical usefulness of some procedures (such as nuclear transfer). However, because of the goal of maintaining all extant gene diversity and because, inevitably, many species are (or will become) 'critically endangered', it is necessary to explore every avenue for a potential contributory role. There are many 'emerging technologies' emanating from the study of livestock and laboratory animals. We predict that a subset of these may have application to the rescue of valuable genes from individual endangered species and eventually to the genetic management of entire populations or species. The present paper reviews the potential candidate techniques and their potential value (and limitations) to the study and conservation of rare wildlife species. PMID- 16478609 TI - [Anatomy and physiology of continence and defecation]. AB - Continence and defecation are two essential functions in humans. Any alteration resulting in anal incontinence and/or constipation can severely impair the patient's quality of life. This study analyzes the anatomical structures and physiologic mechanisms accepted as factors involved in the correct development of both functions, while recognizing that there are still many unclear issues within this complex and sometimes paradoxical structure/function of the human body. PMID- 16478607 TI - Mammalian diversity: gametes, embryos and reproduction. AB - The class Mammalia is composed of approximately 4800 extant species. These mammalian species are divided into three subclasses that include the monotremes, marsupials and eutherians. Monotremes are remarkable because these mammals are born from eggs laid outside of the mother's body. Marsupial mammals have relatively short gestation periods and give birth to highly altricial young that continue a significant amount of 'fetal' development after birth, supported by a highly sophisticated lactation. Less than 10% of mammalian species are monotremes or marsupials, so the great majority of mammals are grouped into the subclass Eutheria, including mouse and human. Mammals exhibit great variety in morphology, physiology and reproduction. In the present article, we highlight some of this remarkable diversity relative to the mouse, one of the most widely used mammalian model organisms, and human. This diversity creates challenges and opportunities for gamete and embryo collection, culture and transfer technologies. PMID- 16478610 TI - [Current utility of endoanal ultrasound in the diagnosis of benign proctological disease]. AB - Endoanal ultrasound is undoubtedly one of the major advances that has taken place in the evaluation of anorectal disease and pelvic floor disorders in the last decade. The main indications for endoanal ultrasound are evaluation of the morphology of the sphincteric apparatus in patients with fecal incontinence, the localization of perianal abscesses and fistulas, the staging of anal cancer and follow-up of squamous cell carcinoma after conservative treatment, and the study and morphological confirmation of lateral internal sphincterotomy in patients with fissure-in-ano, amongst others. In this article we review the main indications and use of endoanal ultrasound in the diagnosis of benign proctological disease. PMID- 16478611 TI - [The surgical treatment of hemorrhoids]. AB - Surgery is the most effective treatment in patients with symptomatic grade III-IV hemorrhoids who have not responded to outpatient treatment, when there is associated abnormalities (anal fissure, anal fistula, skin tags) and in thrombosed hemorrhoids. Hemorrhoidectomy is currently the "gold standard" treatment. Randomized controlled trials comparing open with closed hemorrhoidectomy show no significant differences in pain scores. Stapled hemorrhoidectomy produces less postoperative pain than hemorrhoidectomy but is less effective in terms of symptom control. No treatment is superior to others in reducing postoperative pain except the use of drugs and anesthetic techniques. In patients with prolapsed internal hemorrhoids and thrombosed hemorrhoids, treatment may initially consist of an urgent hemorrhoidectomy with the same results as those obtained with elective surgery. PMID- 16478612 TI - [Treatment of chronic anal fissure]. AB - Current treatment of chronic anal fissure continues to be based on conventional conservative measures in a high percentage of cases. What is known as chemical sphincterotomy aims to achieve a temporary decrease of anal pressures that allows fissures to heal. There are various alternatives such as nitroglycerine or diltiazem ointment and botulinum toxin injections. However, because of collateral effects and recurrences in the medium term, the definitive role of these treatments remains to be elucidated. Nevertheless, chemical sphincterotomy should be the first option in patients with a high risk of incontinence. "Open" or "closed" lateral internal sphincterotomy performed in the ambulatory setting with local anesthesia can currently be considered the ideal treatment of chronic anal fissure refractory to conservative measures so long as the patient is informed about the risk of minor incontinence. This procedure provides rapid and permanent recovery in more than 95% of patients. There is evidence demonstrating that the incontinence rate is related to the extent of the lateral internal sphincterotomy and consequently the extent of this procedure should be reduced to the length of the fissure. PMID- 16478613 TI - [Anal fissure of cryptoglandular origin. Therapeutic options]. AB - Anal fistula is a frequent condition. The most commonly accepted origin is infectious. The most widely used classification is based on cryptoglandular theory and on the position of the fistulous tract in relation to the anal sphincter. Physical examination will help to identify the type of fistula and allow its treatment to be planned. The most widely used complementary tests are endoanal ultrasound and magnetic resonance imaging. We review the various therapeutic options and their results, especially fistulotomy, endorectal advancement flap, use of sedal, anodermal advancement flap, sphincterorrhaphy with sphincter repair, and fibrin glue. PMID- 16478614 TI - [Fecal incontinence. Patient assessment and classical treatments]. AB - Fecal incontinence is a very common disease but its exact prevalence is largely unknown. The condition gives rise to personal and social stigma with severe repercussions for the patient. It can be caused by a large number of physiopathologic disorders and consequently there is a wide variety of treatments. In this article we review the incidence and etiology of fecal incontinence, physical and instrumental examinations (with description of the components of the anorectal laboratory), traditional medical and surgical treatments and, lastly, the results of sphincteroplasties. PMID- 16478615 TI - [Novel techniques in the treatment of anal incontinence]. AB - Fecal incontinence can negatively affect the patient's occupational and social life. Until recently, most patients with severe anal incontinence unresponsive to conservative medical and/or surgical treatments underwent colostomy. Currently, these patients can benefit from one of the innovative techniques that have recently been developed. Thus, the artificial anal sphincter and dynamic graciloplasty are now available, each with specific indications. Both procedures achieve good functional results but complication and reintervention rates are not inconsiderable. Sacral neuromodulation represents an important advance due to its relative simplicity and because, through a period of test stimulation, patients who can definitively benefit from its application can be identified. Other techniques, such as injectable bulking agents or radiofrequency ablation are so recent that experience is limited and their role remains to be defined. Since these techniques are so novel and their economic cost is high, their use should be restricted to study groups with an anorectal physiology laboratory and within the context of clinical trials until experience shows whether or not their application can become widespread. PMID- 16478616 TI - [Rectal prolapse. Abdominal or perineal approach? Current situation]. AB - Rectal prolapse is a major challenge for the surgeon who has to resolve the anatomical problem and the functional disturbances in the same procedure. Abdominal procedures are the most appropriate in young patients, and the most common technique is rectopexia with or without resection. The use of mesh or sutures provides the same results and the choice depends on the surgeon's preference. Laparoscopic surgery has been demonstrated to have similar efficacy to conventional surgery and may become the option of the future. The perineal approach is the best option in elderly patients and in those with associated morbidity; the Delorme technique is simple to carry out, but rectosigmoidectomy provides better results. PMID- 16478617 TI - [Obstructive defecation. Diagnostic methods and treatment]. AB - Obstructive defecation is observed in approximately half of all patients with functional constipation. Functional constipation has been related to alterations in intestinal motility (slow transit constipation) and to pelvic floor disorders leading to obstructive defecation associated with anatomical alterations of the pelvic floor (rectocele, posterior perineal hernia, enterocele and sigmoidocele, internal rectal intussusception, occult mucosal prolapse, solitary rectal ulcer and descending perineum syndrome), or obstructive defecation without anatomical alterations (pelvic floor dyssynergy or anismus). The diagnostic methods used (history and physical examination, colonic transit time, balloon expulsion test, proctography, anorectal manometry and electromyography) are reviewed. Conservative medical treatment and the indications for surgical treatment and its results are also discussed. PMID- 16478618 TI - [Surgical treatment of rectocele and enterocele: an integrated view of disorders of the posterior vaginal compartment]. AB - Genital prolapse, whether associated or not with urinary, anal or sexual dysfunction, should be evaluated globally to select the appropriate treatment. Rectocele and enterocele are defects of the posterior vaginal compartment, although they can be secondary to abnormalities of the central compartment, since lesions of the perineal raphe and rectovaginal septum can occur in isolation or accompanied by others that also affect the tissues involved in pelvic support. The various surgical approaches to rectocele alone or associated with other defects are reviewed. Likewise, the distinct pathogenic types of enterocele are discussed. Laparoscopic sacrocolpoperineopexy is a promising intervention for the simultaneous correction of defects of the posterior and central compartments. New and better designed studies are required to evaluate the distinct surgical approaches and interventions for genital prolapse. PMID- 16478619 TI - Epidemiological risk for Trypanosoma cruzi transmission by species of Phyllosoma complex in the occidental part of Mexico. AB - Domestic and peridomestic triatomine populations were collected in three rural Mexican communities of Jalisco, Nayarit and Zacatecas states. Triatoma longipennis and T. picturata (Phyllosoma complex) were the principal species unequally distributed in the villages: T. longipennis was the main species in two communities and T. picturata in the third one. Peridomestic infestation and colonization indexes were remarkably high ranging from 26.1% to 50% and from 58.3% to 85.7%, respectively. Moreover, domestic (indoor) infestation was observed in only one of the communities infested by T. longipennis. The preliminary study of temporal variation indicates increasing trend of the triatomine population and infestation rates during the dry season. Triatomine infection rates ranged from 41.2% to 60.2% and all the flagellate isolates were assigned to T. cruzi I. The majority of the dwellings were built with modern building materials and the sanitary conditions were generally good. High peridomestic infestations must be considered as a risk factor of Chagas disease transmission and further studies are needed to better understand the peridomestic conditions favoring the establishment of the triatomines. The contribution of such study to enlarger knowledge of epidemiological features of Chagas disease in Mexico is considered. PMID- 16478620 TI - Development of real-time bioradiographic system for functional and metabolic imaging in living brain tissue. AB - We have developed a novel imaging system "real-time bioradiography", which is able to estimate the dynamic changes of physiological function and metabolism in living tissues using positron emitter-labeled tracers and chemiluminescence probes. The apparatus is comprised of a photon-counting camera, image-controller, culturing chamber, reflexible solid scintillator and temperature-controlled imaging chamber. The image distribution of radioactivity and chemiluminescence was acquirable with the reflexible solid scintillator and without, respectively. The reflexible solid scintillator is effective to exclude the affect of intra objective different light reflectivity on radiation detection and to improve the efficiency of radiation detection. To test and to demonstrate the efficacy of this system, we examined the glucose metabolism and superoxide formation during hypoxia-reoxygenation in living brain tissues using 2-[18F]fluoro-2-deoxy-D glucose (FDG) and Lucigenin, respectively. FDG uptake and chemiluminescence images were obtained at time frames of every 15 min. Glucose metabolism was enhanced during the hypoxic treatment, but the superoxide formation was enhanced during reoxygenation. The enhanced glucose metabolism during hypoxia might cause the increase in superoxide formation during reoxygenation. Thus, this new method would open up possibilities to approach simultaneous biological monitoring of a variety of biochemical events with various combinations of positron emitter labeled tracers and chemiluminescence probes in living tissues. PMID- 16478621 TI - The influence of inattention on the neural correlates of scene segmentation. AB - Numerous experiments show that people are unable to report about unattended information. It is also clear that there is extensive processing in the absence of attention. Here, we study, by using an 'inattentional blindness' paradigm while measuring BOLD responses or MEG to texture displays, to what level of scene segmentation visual information is processed when subjects are not capable of reporting that segregating textures were present. We presented non-segregating and occasionally segregating textures in two different conditions: 1. a condition where subjects were not informed about the presence of the segregating textures while at the same time engaged in a foveal task, resulting in prolonged inattentional blindness and 2. a condition with similar task demands, in which, however, the subjects perceived the segregating textures. BOLD responses from early visual areas (V1, V2, V3, V4) and MEG responses up to 240 ms showed a significant difference between segregating and not segregating textures in both conditions and did not differ from each other, indicating that scene segmentation processes proceed normally during inattention. A difference between the two conditions, and hence an influence of attention, was signaled in area V3a and more parietal MEG sensors around 400 ms. PMID- 16478622 TI - The anorectic effect of the selective dopamine D1-receptor agonist A-77636 determined by meal pattern analysis in free-feeding rats. AB - Free-feeding rats meet much of their daily energy requirements by consuming food in meals during the nocturnal phase of the night/day cycle. Meal pattern analysis methodology has been developed to record the patterns of meal taken over a 24-h period, and to provide detailed information on a number of meal-related parameters. Previous work indicates that selective dopamine D1-receptor agonists reduce food intake in short-term feeding tests under the control of homeostatic or hedonic factors. In the present study, our aim was to investigate the effects of the dopamine D1-receptor agonist, A-77636 (0.1-1.0 mg/kg, s.c.), administered just prior to the start of the night period, on the free-feeding and drinking patterns of rats maintained on a standard ad libitum diet. The results indicate that A-77636 exerted a suppressant effect on food intake, due principally to a reduction in meal size and duration. We conclude that there is a dopamine D1 receptor involvement in the normal controls of meal size, and that selective D1 receptor agonists may act to limit meal size. PMID- 16478623 TI - BDNF heterozygous mice demonstrate age-related changes in striatal and nigral gene expression. AB - TrkB receptors mediate the effects of BDNF on striatal medium spiny neurons and mesencephalic dopamine neurons. The effect of partial BDNF gene deletion on locomotor activity and the gene expression of these neurons was evaluated at 3, 12, and 24 months of age in BDNF heterozygous (BDNF(LacZ/neo+)) and wildtype mice. BDNF(LacZ/neo+) mice displayed less spontaneous horizontal activity than wildtypes at 3 and 24 months of age. Whereas striatal preproenkephalin and preprodynorphin mRNA and mesencephalic tyrosine hydroxylase mRNA levels were significantly lower at all ages in BDNF(LacZ/neo+) mice, GAD67 mRNA was only lower at 24 months. In contrast, BDNF(LacZ/neo+) mice expressed more trkB mRNA in the striatum at 3 months and less at 24 months of age than wildtypes. Total striatal cell number in the two genotypes was not different at 12 months of age, whereas Golgi staining revealed that the spine density on distal dendrites of medium spiny neurons was less in BDNF(LacZ/neo+) mice than in wildtypes at 24 months of age. These data indicate that endogenous BDNF is required to maintain the normal phenotype and functioning of striatal projection neurons and mesencephalic dopamine neurons and that exaggerated dysfunction of these neurons and a concomitant decline in locomotor behavior occurs during aging. PMID- 16478624 TI - Increases in the activated forms of ERK 1/2, p38 MAPK, and CREB are correlated with the expression of at-level mechanical allodynia following spinal cord injury. AB - Rats given moderate spinal cord injury (SCI) display increases in the expression of the activated form of the transcription factor cyclic AMP responsive element binding protein (CREB) in spinal segments of dermatomes corresponding to permanent mechanical allodynia, a model of chronic central neuropathic pain (CNP; (Crown, E.D., Ye, Z., Johnson, K.M., Xu, G.Y., McAdoo, D.J., Westlund, K.N., Hulsebosch, C.E., 2005. Upregulation of the phosphorylated form of CREB in spinothalamic tract cells following spinal cord injury: relation to central neuropathic pain. Neurosci. Lett. 384, 139-144)). Given that not all rats that receive moderate SCI develop CNP, the current study was designed to further analyze changes in persistent CREB activation and in the activation state of upstream intracellular signaling cascades (e.g., mitogen-activated protein kinases [MAPKs]) in populations of rats that receive SCI and weeks later develop CNP and rats that receive SCI but do not develop CNP. The results indicate that activated kinases such as pERK 1/2, p-p38 MAPK, but not pJNK, are upregulated in injured rats that develop CNP as compared to injured rats that fail to develop CNP. In addition, the current results replicated our previous finding that activated CREB is upregulated following SCI, however, only in SCI rats that developed CNP. Taken together, these results indicate that activation of intracellular signaling cascades traditionally associated with long-term potentiation and memory is associated with the expression of chronic CNP following SCI. PMID- 16478625 TI - On-line sequential preconcentration of inorganic anions by anion-selective exhaustive injection and base-stacking in capillary zone electrophoresis. AB - A sequential electrostacking method based on anion-selective exhaustive injection (ASEI) and base-stacking (BS) is presented for the preconcentration and determination of inorganic anions by capillary zone electrophoresis (CZE) in this paper. Tetradecyltrimethylammonium bromide as an electroosmotic flow (EOF) modifier was added into the buffer to suppress EOF of the capillary. Firstly, a water plug was hydrodynamically injected into the capillary. During ASEI under negative high voltage, the sample anions migrated quickly towards the boundary between the water plug and buffer in the capillary. Then an alkaline zone was injected electrokinetically to concentrate the anions further. With the sequential electrostacking method, the preconcentration factor of (0.8-1.3) x 10(5) was obtained compared with the conventionally electrokinetic injection and the relative standard deviation of peak area was 3.3-5.3% (n = 5). The detection limits of ASEI-BS-CZE for six inorganic anions were 6-14 ng/L. The proposed method has been adopted to analyze six anions in cigarette samples successfully. PMID- 16478626 TI - Transcranial magnetic stimulation screening for cord compression in cervical spondylosis. AB - OBJECTIVE: Cervical spondylosis (CS) often results in various degrees of cord compression, which can be evaluated functionally with transcranial magnetic stimulation (TMS). We investigate the use of TMS as a screening tool for myelopathy in CS. METHODS: We prospectively studied 231 patients classified into Groups 1 to 4 based on MRI grading of severity of cord compromise. TMS elicited central motor conduction times and motor evoked potential (MEP) amplitudes in all 4 limbs. The results were compared with those from 45 healthy controls. RESULTS: TMS showed 98% sensitivity and 98% specificity for cord abnormality using MRI as reference standard. CONCLUSIONS: MEP abnormalities are useful for electrophysiological evaluation of cord compression in CS. While TMS is not a substitute for MRI, it is of value as a rapid, inexpensive and non-invasive technique for screening patients before MRI studies. PMID- 16478627 TI - Effect of intrauterine position on sex differences in the gabaergic system and behavior of rats. AB - In multiparous species such as the rat (in this case the albino Wistar strain), steroid influence during fetal growth is affected by the relative intrauterine position of male and female fetuses and is stronger when the potential effects of contiguity and caudal position are combined. The effect of intrauterine position on gonadal steroid levels in neonatal and adult animals was examined using radioimmunoassay techniques. Since the organizing effect of prenatal steroids may influence the postnatal GABA content, HPLC was used to determine the gabaergic content in several hypothalamic and limbic areas in the adult rat. The effects of intrauterine position on adaptive behavior were examined by recording exploratory behavior (using the corridor and hole board tests) and intraspecific aggression (induced by isolation). Female pups influenced by males during development produced more testosterone. In adult males, those that developed closer to the cervix (and with no influence from other fetuses) produced more testosterone and less estradiol. The same animals also produced more hypothalamic GABA and showed greater exploratory capacity. No significant differences were seen between any experimental groups with respect to aggression. These results show increased variability between males with respect to adult exploratory behavior, and in the neurochemical and endocrine systems involved, due to intrauterine position during development. The effect of this physiological phenomenon on the structure of rodent populations is discussed. PMID- 16478628 TI - Combined administration of barbourin--albumin and hirudin--albumin fusion proteins limits fibrin(ogen) deposition on the rabbit balloon-injured aorta. AB - INTRODUCTION: There are continuing needs for new antithrombotic agents and procedures. We hypothesized that the slowly cleared recombinant fusion proteins barbourin--albumin (BLAH6) and hirudin--albumin (HLAH6) would be effective in limiting fibrin(ogen) and/or platelet deposition in a rabbit model of arterial injury. MATERIALS AND METHODS: Recombinant fusion proteins were expressed in Pichia pastoris fermenter cultures and purified by nickel-chelate affinity chromatography. They were injected intravenously into rabbits prior to blood sampling and platelet aggregometry, assessment of deposition of 125I-fibrin(ogen) and 51Cr-platelet onto the balloon-injured thoracic aorta, electron microscopy (EM) and immunohistochemistry of aortic sections, and determination of bleeding time following a standardized ear incision. RESULTS AND CONCLUSIONS: BLAH6 administration elicited a dose- and time-dependent inhibition of platelet aggregation in post-injection whole blood samples, and reduced both fibrin(ogen) and platelet deposition on the injured aorta, although the former effect was both more durable and more significant than the latter. In contrast, HLAH6 injection reduced fibrin(ogen) but not platelet deposition. Doses of the two proteins ineffective in preventing fibrin(ogen) deposition when given alone were effective when combined, suggesting at least additive effects. Immunohistochemistry and EM supported the radioactive deposition studies, while bleeding times were decreased with combined BLAH6 and HLAH6 administration compared to HLAH6 alone in a rabbit ear bleeding model. The data show that these fusion proteins exert an antithrombotic effect in vivo and may indicate that combined low-dose administration of antiplatelet and antithrombin agents could offer safety advantages in the treatment of thrombosis. PMID- 16478629 TI - Intraarterial cisplatin/nedaplatin and intravenous 5-fluorouracil with concurrent radiation therapy for patients with high-risk uterine cervical cancer. AB - OBJECTIVE: The purpose of this study was to determine the effectiveness of the combination of intraarterial and intravenous concurrent chemoradiation therapy (CIAIV-CCRT) for the treatment of high-risk uterine cervical cancer. METHODS: Between January 2000 and November 2004, we reviewed 45 cervical cancer patients treated by CIAIV-CCRT. The numbers of patients with stage IB2, IIA, IIB, IIIA, IIIB, and IVA were 3, 6, 14, 1, 17, and 4, respectively. Patients with stage III and IVA or patients with tumors >3 cm in diameter were enrolled in this study. Two sessions of CCRT were administered every 3 weeks using a combination of 70 mg/m2 x h(-1) cisplatin or 50 mg/m2 x h(-1) nedaplatin via the bilateral uterine artery and 2800 mg/m2 x 96 h(-1) 5-fluorouracil intravenously. Patients concurrently received external beam radiation therapy and brachytherapy. A nonrandomized control group of 47 patients who underwent radiation therapy alone between 1993 and 2000 was used for comparison. RESULTS: Of the 45 patients, 28 (62%) exhibited complete response and 16 (36%) exhibited partial response. One IIIB patient (2%) did not show any response. The 5-year overall survival (OAS) rates in the CCRT group and control group were 80.6% and 54.9%, respectively. With regard to late toxicities, no statistically significant differences were observed between the two groups. In uni- and multivariate analyses, positive pelvic lymph node showed a statistically significant influence on the OAS in the CIAIV-CCRT group (P = 0.049). CONCLUSION: These preliminary results suggest that CIAIV-CCRT can improve the prognosis of patients with high-risk cervical cancer. PMID- 16478630 TI - Evaluation of pegylated liposomal doxorubicin (Doxil) as second-line chemotherapy of squamous cell carcinoma of the cervix: a phase II study of the Gynecologic Oncology Group. AB - OBJECTIVE: Doxorubicin has reported activity in advanced and recurrent cervical cancer but hematologic toxicity has limited its use in some combinations. To determine the level of activity and potential for use in future combinations, a phase II trial of pegylated liposomal doxorubicin as second-line therapy in advanced and recurrent cervical cancer was performed. METHODS: Eligible patients had squamous cell carcinoma of the cervix, measurable disease, one prior chemotherapy regimen which did not include an anthracycline, absolute neutrophil count (ANC) > 1500/microl, platelet count > 100,000/microl, and adequate hepatic function. Pegylated liposomal doxorubicin 40 mg/m(2) was administered intravenously over 1 h every 4 weeks. RESULTS: Twenty-seven patients were entered on this study. All patients were evaluable for toxicity and 26 were evaluable for response. A median of 2 courses of therapy (range 1-10) was given. No grade 4 toxicities were noted. Three patients (11.1%) had partial responses. CONCLUSION: Liposomal doxorubicin has limited activity, at the dose and schedule employed in previously-treated cervical cancer. PMID- 16478631 TI - Triapine and cytarabine is an active combination in patients with acute leukemia or myelodysplastic syndrome. AB - Triapine, an iron chelator and a potent inhibitor of ribonucleotide reductase, has significant anti-leukemia activity. A phase I study of Triapine in combination with ara-C was conducted in 32 patients with refractory acute leukemia and high-risk MDS. Triapine (105 mg/m2/day 6-h infusion) was followed immediately by ara-C [100 (n=4), 200 (n=6), 400 (n=7), or 800 (n=8)mg/m2/day] as an 18-h infusion for 5 consecutive days. Dose-limiting toxicities (DLTs) were observed at the 800 mg/m2 ara-C dose level (one patient each with grade 4 mucositis; grade 4 neutropenic colitis, sepsis; grade 4 neuropathy; and grade 4 hyperbilirubinemia). Therefore, the study was amended to include an ara-C dose level of 600 mg/m2/day, no DLTs occurred in seven patients treated at this dose level. Mean Triapine C(max) and AUC were 1.13 microg/mL and 251.5 minmicrog/mL. Of 31 evaluable patients, 4 (13%) (3 AML, 1 Ph+ALL) achieved a CR (1 at a dose of 800 mg/m2; 2 at 600 mg/m2; 1 at 200mg/m2). The recommended phase II regimen is Triapine 105 mg/m2/day followed by ara-C 600 mg/m2/day for 5 consecutive days every 3-6 weeks. PMID- 16478632 TI - Cloning and expression analysis of interferon-gamma-inducible-lysosomal thiol reductase gene in large yellow croaker (Pseudosciaena crocea). AB - In mammals, interferon-gamma-inducible-lysosomal thiol reductase (GILT) has been demonstrated to play a key role in the processing and presentation of MHC class II-restricted antigen (Ag) by catalyzing disulfide bond reduction, thus unfolding native protein Ag and facilitating subsequent cleavage by proteases. Here, we reported the cloning of a GILT gene homologue from the spleen of large yellow croaker, a marine fish (LycGILT). The full-length cDNA of LycGILT gene is 1033 nucleotides (nt) encoding a protein of 256 amino acids (aa), with a putative molecular weight of 28.9 kDa. The deduced protein is highly homologous to that of mammalian and zebrafish GILTs and shares 54.1% sequence identity to that of zebrafish and 43.2-39.2% sequence identity to that of various mammals. The deduced LycGILT possesses the typical structural feature of mammalian GILT, including an active-site CXXC motif, a GILT signature sequence CQHGX2ECX2NX4C, and other six cysteines responsible for the formation of disulfide bonds in the C terminus. Genomic analysis revealed that LycGILT gene, spanning a 3159nt fragment, contained seven exons interrupted by six introns and exhibited a similar exon-intron organization to human and mouse GILT genes except for a slightly more compact intron arrangement. The LycGILT expression is obviously up regulated in spleen and kidney after immunization with inactivated trivalent bacterial vaccine consisting of Vibrio alginolyticus, V. paraphaemolyticus, and Aeromonas hydrophila although it also is constitutively expressed in liver, gills, brain, and heart, suggesting that LycGILT may be involved in the immune response to bacterial challenge in large yellow croaker. A search of NCBI sequence data with LycGILT cDNA identified a pufferfish (fugu rubrides) GILT homologue cDNA and its genomic DNA sequence, where two putative interferon-gamma activation sites (GAS) were found within the promoter region. This provided evidence that a fish GILT homologue like mammalian GILT, may also be regulated by interferon-gamma (IFN-gamma) through the JAK-STAT signal pathway. These results indicate that the bony fish GILT is a functional homologue of mammalian GILT. PMID- 16478633 TI - Psychosocial functioning of adolescents with and without paediatric bipolar disorder. AB - BACKGROUND: While there has been a growing interest in the presentation of Bipolar Disorder (BD) in children and adolescents, few studies have investigated the psychosocial functioning of these individuals and its relationship to trauma and suicidal ideation. METHODS: 63 adolescents aged 13-17 participated: 39 controls and 24 with Bipolar Disorder (BD). Group allocation and histories of trauma and suicidal ideation were obtained using the K-SADS-PL and WASH-U-KSADS. Adolescents completed questionnaires covering negative life events, self-esteem, hopelessness, regulation of anger, locus of control and coping. RESULTS: More traumatic events and negative life experiences were reported by the BD group with over 50% of the BD sample indicating a history of trauma compared with 10% of the controls. The BD group reported lower self-esteem, more hopelessness, more negative life events, a more external locus of control and greater difficulties regulating emotion in anger-provoking situations. They were also found to have poorer coping strategies than the controls. Histories of trauma did not differentiate those with and without psychosocial problems. Further, hopelessness was found to be the best predictor of those BD adolescents reporting suicidal ideation. Comorbidity could not account for the differences found. LIMITATIONS: The sample was small and therefore disallowed comparisons among subtypes of BD. Cross-sectional design limited the ability to investigate causal relationships. CONCLUSIONS: This is the first study to document the widespread psychosocial difficulties facing youth with BD, highlighting these issues as important ones to explore during assessment and treatment, particularly in management of affective storms and suicidal risk. PMID- 16478634 TI - Construction of hybrib Autographa californica nuclear polyhedrosis bacmid by modification of p143 helicase. AB - We developed a new hybrid nuclear polyhedrosis virus (NPV) bacmid capable of infecting Spodoptera frugiperda, Tricoplusia ni, and Bombyx mori, and B. mori cell lines for producing hybrid recombinant baculovirus that can carry a gene of interest and express it in a broad range of hosts. A GFP(uv)-beta1,3-N acetylglucosaminyltransferase 2 fusion gene was expressed successfully in silkworm larvae using this hybrid bacmid. The hybrid NPV bacmid provides an altogether simple and realistically feasible method for large-scale applications using silkworm larvae. It can be easily managed in E. coli, which has no biohazard safety concerns, in addition to the baculovirus-based expression system. PMID- 16478635 TI - Hepatocyte nuclear structure and subcellular distribution of copper in zebrafish Brachydanio rerio and roach Rutilus rutilus (Teleostei, Cyprinidae) exposed to copper sulphate. AB - Copper is a trace element essential to life, but also a heavy metal with toxic effect clearly demonstrated. Cu induced perturbations in fish liver are well documented but the variability of the reported results is large. In this study two cyprinids, zebrafish and roach, were exposed to copper. Reported histocytological changes are either adaptative or degenerative depending on fish species, concentration of metal, and duration of exposure. Hepatic subcellular distribution of copper was determined by X-ray microanalysis in control and Cu exposed roach and zebrafish. Sublethal copper sulphate contamination induced the development of a particular nucleolar alteration forming a network or honeycomb like structure in liver. This perturbation is observable in almost all the hepatocytes of zebrafish and roach exposed to copper for a minimum of 4 days of exposure. It seemed to concern more precisely the pars fibrosa. X-ray microanalysis showed that the appearance of network nucleolus was in relation to a Cu accumulation. Cu deposit was well located in the network as pars granulosa and nucloplasm showed very lower metal concentrations. The origin and consequence of network structure in nucleolus was discussed. PMID- 16478636 TI - The effect of slaughter operations on the contamination of chicken carcasses with thermotolerant Campylobacter. AB - To evaluate the effect of specific slaughter operations on the contamination of broiler carcasses with naturally occurring thermotolerant Campylobacter, experiments were carried out in two Danish commercial slaughter plants (Plant I and Plant II). Six broiler flocks determined Campylobacter positive prior to slaughter were investigated at four sampling locations within each slaughter plant. Quantification of thermotolerant Campylobacter in 30 neck skin samples per flock per sampling location showed that the evisceration operation in Plant I led to a significant increase in the Campylobacter concentration of 0.5 log(10) cfu/g in average, whereas no significant changes were observed during this operation in Plant II. Air chilling (Plant I) and water chilling (Plant II), both including a carcass wash prior to the chilling operation, caused similar, but significant reductions of 0.83 and 0.97 log(10) cfu/g, respectively. In packed frozen chickens (Plant II) an additional reduction of 1.38 log(10) cfu/g in average was obtained due to the freezing operation. In packed chilled chickens (Plant I), however, the number of thermotolerant Campylobacter per gram remained at the same level as after air chilling. Enumeration of thermotolerant Campylobacter in 30 intestinal samples per flock showed that in two of the six flocks examined the within flock colonization was very low (<3% and 27% positive samples). The remaining four flocks were colonized at percentages of 100 (three flocks) and 97 (one flock) and had intestinal mean counts ranging from 6.65 to 8.20 log(10) cfu/g. A correlation between Campylobacter concentrations in intestinal content and on chicken carcasses after the defeathering operation was documented. This finding indicates that a reduction in the Campylobacter concentration on chicken carcasses may also be obtained by interventions aimed at reducing the concentration of Campylobacter in the intestines of the living birds. PMID- 16478637 TI - Viability of commercial probiotic cultures (L. acidophilus, Bifidobacterium sp., L. casei, L. paracasei and L. rhamnosus) in cheddar cheese. AB - Six batches of cheddar cheese were manufactured containing different combinations of commercially available probiotic cultures from three suppliers. Duplicate cheeses contained the organisms of each supplier, a Bifidobacterium spp. (each supplier), a Lactobacillus acidophilus (2 suppliers), and either Lactobacillus casei, Lactobacillus paracasei, or Lactobacillus rhamnosus. Using selective media, the different strains were assessed for viability during cheddar cheese maturation over 32 weeks. The Bifidobacterium sp. remained at high numbers with the three strains being present in cheese at 4 x 10(7), 1.4 x 10(8), and 5 x 10(8) CFU/g after 32 weeks. Similarly the L. casei (2 x 10(7) CFU/g), L. paracasei (1.6 x 10(7) CFU/g), and L. rhamnosus (9 x 10(8) CFU/g) strains survived well; however, the L. acidophilus strains performed poorly with both decreasing in a similar manner to be present at 3.6 x 10(3) CFU/g and 4.9 x 10(3) CFU/g after 32 weeks. This study indicates that cheddar cheese is a good vehicle for a variety of commercial probiotics but survival of L. acidophilus strains will need to be improved. PMID- 16478638 TI - Effect of acacia (Robinia pseudo-acacia L.) honey on the characteristic microflora of yogurt during refrigerated storage. AB - The primary purpose of this research was to monitor the influence of acacia honey addition to yogurt milk on survival of the microbial flora of yogurt during refrigerated storage for 6 wk. Results showed that the presence of honey at 1.0% to 5.0% (w/v) did not significantly influence (P>0.05) the viability of characteristic microorganisms (i.e., Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus) in yogurt during storage at 4 degrees C. Similarly, honey had no effect on pH and lactic acid levels of the final products. Despite these findings, enrichment of yogurt with honey is recommended because honey is a natural sweetener that possesses a wide range of beneficial nutritional properties. In addition, at a rate of approximately 3.0% (w/v), it highly improves the sensory quality of the finished product without having a detrimental effect on characteristic lactic acid bacteria. PMID- 16478639 TI - Application of gold nanoparticles to microencapsulation of thioridazine. AB - Thioridazine-containing ethyl cellulose (EC) microcapsules were prepared in the presence of gold nanoparticles via the W/O/W emulsification solvent-evaporation method. The gold nanoparticles have been verified as human safe and the nondestructive physisorption of thioridazine on gold nanoparticles was corroborated with the time-of-flight second ion mass spectrometry measurements. The morphology of the formed microcapsules (ETA, containing EC, Thioridazine and Au) changed substantially because of the presence of gold nanoparticles. In addition to a prolonged controlled release, these ETA microcapsules had an enhanced thioridazine encapsulation with an efficiency over one and half times that of the microcapsules (ET) containing no nanogold particles. While data of the release kinetics for ET microcapsules fitted the apparent first-order model, corresponding data for ETA microcapsules agreed better with the Higuchi model indicating a uniform distribution of thioridazine in the monolithic-type microcapsules. PMID- 16478640 TI - Induction of cancer cell-specific apoptosis by folate-labeled cationic liposomes. AB - We have previously reported that cationic liposomes themselves can induce apoptosis in macrophages and lymphocytes. In this paper, we attempted the cancer cell-specific delivery of cationic liposomes and the induction of apoptosis utilizing this characteristic. Cationic liposomes composed of stearylamine (SA liposomes) induced apoptosis in human nasopharyngeal epidermoid carcinoma cells (KB cells) overexpressing the folate receptor and human fibroblasts (WI-38 cells) with no folate receptor, without showing selectivity. To recruit liposomes to cancer cells and induce apoptosis, we focused on the folate receptor and prepared folic acid-labeled liposomes using polyethyleneglycol (PEG) (folate-PEG liposomes). Folate-PEG-liposomes showed selectivity and induced apoptosis in KB cells, but not WI-38 cells. The apoptosis occurred in a dose-dependent manner. Furthermore, folate-PEG-liposomes appear to associate with KB cells via the folate receptor, whereas SA-liposomes may associate with cells through electrostatic interactions. To confirm the contribution of the folate receptor to apoptosis of KB cells induced by folate-PEG-liposomes, the effect of folic acid on the apoptosis was examined. The addition of free folic acid drastically suppressed the apoptosis of KB cells and the percentage of cells with hypodiploid nuclei returned to the control level. Taken together, cationic liposomes labeled with folate bound to KB cells via folate receptors and, interestingly, the cationic liposomes themselves could cause apoptosis in cancer cells. PMID- 16478641 TI - Eve-3: a liver enriched suppressor of Ras/MAPK signaling. AB - BACKGROUND/AIMS: The developed liver is able to tightly control cellular proliferation, rapidly switching from quiescence to growth in response to specific stimuli. This suggests that growth inhibitors may be involved in the control of liver growth. We analyzed the role of the Spred-family of growth inhibitors in the liver. METHODS: We screened human EST databases for Spred related sequences. Clones were isolated, sequenced, epitope-tagged and expressed. Subcellular localization of clones were determined and their effects on cellular signaling pathways analysed using specific antibodies. Cell cycle progression assays and protein interaction studies were initiated. Organ distribution of transcripts and their expression throughout liver development and in primary hepatocytes were recorded. RESULTS: We have identified a new, liver-restricted protein, Eve-3, containing a single Ena Vasp homology (EVH1) domain that can potently block activation of the Ras/MAPK pathway. Eve-3 is specific in inhibiting the Ras/MAPK pathway. Eve-3 can block serum-mediated cell cycle progression and its expression is highly regulated during liver development. CONCLUSIONS: The liver is the only organ that can regulate its growth and mass. Eve-3 may act as an inhibitor of proliferation pathways in the mature liver and be involved in modulating the unique regenerative capacity of this organ. PMID- 16478642 TI - Analysis of EEG tracings in frequency and time domain in hepatic encephalopathy. AB - Spectral EEG analysis in hepatic encephalopathy (HE) is usually performed disregarding the effect of epoch length, statistical errors and equipment noise. A study on these items was carried out. In addition, spectral analysis and a new analysis, performed in time domain, were compared in the assessment of HE. The EEG tracings of 73 cirrhotic patients with HE were analyzed. Artifact-free periods of about 1 min were selected. Equipment noise was measured by short circuiting all the electrodes. The equipment noise was notable below 1.5 Hz; the best epoch length was 4s and the statistical errors were minimal for the band with the highest relative power. Nineteen per cent of the tracings were unstable. The spectral values were found to be related to liver function and to the degree of HE, whereas the relationship with psychometric variables was poor. The indexes computed by time-domain analysis were found to be better related to psychometric findings. We have provided information on the optimisation of spectral EEG analysis and presented a time-domain analysis giving results related to psychometric tests and liver function. PMID- 16478643 TI - Should chemotherapy combinations for advanced non-small cell lung cancer be platinum-based? A meta-analysis of phase III randomized trials. AB - BACKGROUND: Non-platinum regimens have been proposed as an alternative to the platinum-based combinations for treatment of advanced non-small cell lung cancer. However, conflicting results were reported. METHODS: Meta-analysis of phase III trials randomizing platinum-based versus non-platinum combinations as first-line chemotherapy with 1-year survival rate as a primary endpoint. Fourteen trials have been identified. Experimental arms were gemcitabine/vinorelbine (n=4), gemcitabine/taxane (n=7), gemcitabine/epirubicin (n=1), paclitaxel/vinorelbine (n=1), and gemcitabine/ifosfamide (n=1). The comparator was a doublet of a platinum compound plus a third generation agent for all but two studies (triplets). Updated data were available for 13 studies. The Peto and Yusuf method was used to generate odds ratios (OR). All tests are two-sided. RESULTS: A statistical heterogeneity was detected when the 13 studies were analyzed. Considering that current guidelines recommend platinum-based doublets as standard therapy we therefore limited the meta-analysis to the set of 11 phase III studies which used a platinum-based doublet (2298 and 2304 patients in platinum-based and non-platinum arms, respectively). No significant heterogeneity was detected in this consistent group of studies. Patients treated with a platinum-based regimen benefited from a statically significant reduction in the risk of death at 1 year (OR: 0.88, 95% CI 0.78-0.99; p=0.044) and a lower risk of being refractory to chemotherapy (OR: 0.87, 0.73-0.99; p=0.049). Forty-four (1.9%) and 29 (1.3%) toxic-related deaths were reported for platinum-based and non-platinum regimens, respectively (OR: 1.53; 0.96-2.49, p=0.08). An increased risk of grade 3-4 gastro intestinal and hematological toxicity for patients receiving platinum-based chemotherapy was statistically demonstrated. There was no statically significant increase in risk of febrile neutropenia, OR=1.23 (0.94-1.60, p=0.063). CONCLUSION: A platinum-based doublet induced a statically significant reduction in the risk of death when compared with non-platinum chemotherapy without inducing an unacceptable increase in toxicity. PMID- 16478644 TI - Sensitivity of bacterial biofilms and planktonic cells to a new antimicrobial agent, Oxsil 320N. AB - The effective concentrations of disinfectants were determined for planktonic bacteria using the norms EN 1040 and NF T 72-150. This concentration corresponds to biocide efficacy after 5 min of contact, followed by neutralization. However, micro-organisms often colonize a substratum and form microcolonies or biofilms where they are enclosed in exopolymer matrices. Biofilms are commonly resistant to a broad range of antimicrobial agents, and resistance mechanisms involve exopolymer matrices, changes in gene expression and metabolic alterations. Due to these different resistance mechanisms, it is difficult to select and titrate antimicrobial agents to be effective against biofilms. In this context, SODIFRA developed a new disinfectant, Oxsil 320N (French patent 94 15 193). Oxsil 320N is an association of three active principles: hydrogen peroxide, acetic acid/peracetic acid and silver. This biocide was tested on planktonic bacteria and on 24-h biofilms formed on AISI 304 stainless steel surfaces. The effective concentration of Oxsil 320N was also determined on biofilms using SODIFRA recommendations (without neutralization of the biocide). Data showed that the antimicrobial efficacy measured on planktonic bacteria is not a reliable indicator of performance when biofilm is present. When biofilms were exposed to Oxsil 320N, the concentration needed to achieve a 10(5)-fold decrease in concentration was 10 times higher than that for bacterial suspensions (0.313% Oxsil 320N). An effective concentration of Oxsil 320N of 3.13% was required. PMID- 16478645 TI - Handwashing: a simple, economical and effective method for preventing nosocomial infections in intensive care units. AB - As most nosocomial infections are thought to be transmitted by the hands of healthcare workers, handwashing is considered to be the single most important intervention to prevent nosocomial infections. However, studies have shown that handwashing practices are poor, especially among medical personnel. This review gives an overview of handwashing in health care and in the community, including some aspects that have attracted little attention, such as hand drying and cultural issues determining hand hygiene behaviour. Hand hygiene is the most effective measure for interrupting the transmission of micro-organisms which cause infection, both in the community and in the healthcare setting. Using hand hygiene as a sole measure to reduce infection is unlikely to be successful when other factors in infection control, such as environmental hygiene, crowding, staffing levels and education, are inadequate. Hand hygiene must be part of an integrated approach to infection control. Compliance with hand hygiene recommendations is poor worldwide. While the techniques involved in hand hygiene are simple, the complex interdependence of factors that determine hand hygiene behaviour makes the study of hand hygiene complex. It is now recognized that improving compliance with hand hygiene recommendations depends on altering human behaviour. Input from behavioural and social sciences is essential when designing studies to investigate compliance. Interventions to increase compliance with hand hygiene practices must be appropriate for different cultural and social needs. PMID- 16478646 TI - Disruption of transforming growth factor beta-Smad signaling pathway in head and neck squamous cell carcinoma as evidenced by mutations of SMAD2 and SMAD4. AB - The role of the TGF-beta-Smad signaling pathway in the carcinogenesis of head and neck cancer has not been fully evaluated genetically. In this study, we screened for mutation in the five main members of the TGF-beta -Smad signaling pathway, TGF-beta type I receptor (TGFBRI), TGF-beta type II receptor (TGFBRII), SMAD2, SMAD3 and SMAD4, in eight human head and neck squamous cell carcinoma (HNSCC) cell lines. Two mutations with presumed loss of heterozygosity (LOH) were identified. A novel missense mutation of SMAD2, located in exon 8 at codon 276 TCG (ser) -->TTG (leu), was identified in cell line SCC-15. This is the first report of a biallelic mutation of the SMAD2 gene in HNSCC. A nonsense mutation of the SMAD4 gene in exon 5 codon 245 CAG (glut) -->TAG (stop) was found in cell line CAL27. Western blotting verified that this nonsense mutation gives rise to the complete loss of the Smad4 protein in the cells. While the down-regulation and loss of expressions of the TGF-beta-Smad signaling pathway have been described frequently in HNSCC, here we offer further genetic evidence that the pathway is directly targeted for mutation during the HNSCC tumorigenesis. PMID- 16478647 TI - Soluble TRAIL gene and actinomycin D synergistically suppressed multiple metastasis of TRAIL-resistant colon cancer in the liver. AB - Metastatic liver tumors are highly malignant and refractory to conventional therapies. TRAIL-resistant CT-26 cells underwent apoptosis in vitro in the presence of both recombinant TRAIL (rTRAIL) and a suboptimal dose of actinomycin D (ACD). Co-administration of soluble TRAIL (sTRAIL) gene and ACD suppressed the metastatic liver tumors of CT-26, significantly inducing apoptosis in the tumors, while such effects were not demonstrated in mice that received either the sTRAIL gene or ACD alone. The gene therapy of sTRAIL with a suboptimal dose of an anticancer drug is a new strategy for treatment of multiple liver metastasis. PMID- 16478648 TI - A simple procedure for calculating condensed phase heat of formation of nitroaromatic energetic materials. AB - A simple procedure has been introduced for calculating solid and liquid heats of formation of nitroaromatic energetic compounds. This model assumes that the heat of formation of a nitroaromatic compound of composition C(a)H(b)N(c)O(d) can be expressed as a new correlation which depends on elemental composition and various structural and special functional group parameters. Condensed phase heats of formation predicted using the method described herein and complicated quantum mechanical computations [B.M. Rice, J. Hare, Thermochem. Acta 384 (2002) 377] have a root mean square (rms) deviation of 5.9 and 11.1 kcal/mol for 19 well known organic nitroaromatic compounds. Predicted heats of formation for 29 polycyclic nitroaromatic energetic compounds have a rms deviation from experiment of 10.6 kcal/mol. The results show that the present method gives comparable prediction respect to the other methods such as complex quantum mechanical computation. PMID- 16478649 TI - Galectin-3: an open-ended story. AB - Galectins, an ancient lectin family, are characterized by specific binding of beta-galactosides through evolutionary conserved sequence elements of carbohydrate-recognition domain (CRD). A structurally unique member of the family is galectin-3; in addition to the CRD it contains a proline- and glycine-rich N terminal domain (ND) through which is able to form oligomers. Galectin-3 is widely spread among different types of cells and tissues, found intracellularly in nucleus and cytoplasm or secreted via non-classical pathway outside of cell, thus being found on the cell surface or in the extracellular space. Through specific interactions with a variety of intra- and extracellular proteins galectin-3 affects numerous biological processes and seems to be involved in different physiological and pathophysiological conditions, such as development, immune reactions, and neoplastic transformation and metastasis. The review attempts to summarize the existing information on structural, biochemical and intriguing functional properties of galectin-3. PMID- 16478650 TI - Molecular and bioinformatic analysis of the FB-NOF transposable element. AB - The Drosophila melanogaster transposable element FB-NOF is known to play a role in genome plasticity through the generation of all sort of genomic rearrangements. Moreover, several insertional mutants due to FB mobilizations have been reported. Its structure and sequence, however, have been poorly studied mainly as a consequence of the long, complex and repetitive sequence of FB inverted repeats. This repetitive region is composed of several 154 bp blocks, each with five almost identical repeats. In this paper, we report the sequencing process of 2 kb long FB inverted repeats of a complete FB-NOF element, with high precision and reliability. This achievement has been possible using a new map of the FB repetitive region, which identifies unambiguously each repeat with new features that can be used as landmarks. With this new vision of the element, a list of FB-NOF in the D. melanogaster genomic clones has been done, improving previous works that used only bioinformatic algorithms. The availability of many FB and FB-NOF sequences allowed an analysis of the FB insertion sequences that showed no sequence specificity, but a preference for A/T rich sequences. The position of NOF into FB is also studied, revealing that it is always located after a second repeat in a random block. With the results of this analysis, we propose a model of transposition in which NOF jumps from FB to FB, using an unidentified transposase enzyme that should specifically recognize the second repeat end of the FB blocks. PMID- 16478652 TI - Satoyoshi syndrome: a rare multisystemic disorder requiring systemic and symptomatic treatment. AB - Satoyoshi syndrome is a rare multisystemic disorder with assumed autoimmune pathogenesis. Typical clinical features are progressive painful muscle spasms, alopecia, diarrhoea, and skeletal and endocrine abnormalities often resulting in early invalidism and death. Patients have been treated with immunoglobulins and glucocorticoids with varying outcome. We report on a 19-year-old German adolescent who has been successfully treated with a new combination of carbamazepine to reduce the severity and frequency of painful nocturnal muscle spasms, prednisolone, methotrexate and sex-steroids. Prednisolone treatment alone was not successful. After introduction of low-dose of methotrexate to the therapy the patient recovered from muscle spasms, alopecia and diarrhoea. Initiation of sex-steroid treatment resulted in pubertal development, regular menstrual cycles and improved quality of life. PMID- 16478651 TI - Higher activity of polymorphic NAD(P)H:quinone oxidoreductase in liver cytosols from blacks compared to whites. AB - In human liver, the two-electron reduction of quinone compounds, such as menadione is catalyzed by cytosolic carbonyl reductase (CBR) and NAD(P)H:quinone oxidoreductase (NQO1) activities. We assessed the relative contributions of CBR and NQO1 activities to the total menadione reducing capacity in liver cytosols from black (n=31) and white donors (n=63). Maximal menadione reductase activities did not differ between black (13.0+/-5.0 nmol/min mg), and white donors (11.4+/ 6.6 nmol/min mg; p=0.208). In addition, both groups presented similar levels of CBR activities (CBR(blacks)=10.9+/-4.1 nmol/min mg) versus CBR(whites)=10.5+/-5.8 nmol/min mg; p=0.708). In contrast, blacks showed higher NQO1 activities (two fold) than whites (NQO1(blacks)=2.1+/-3.0 nmol/min mg versus NQO1(whites)=0.9+/ 1.6 nmol/min mg, p<0.01). To further explore this disparity, we tested whether NQO1 activity was associated with the common NQO1(*)2 genetic polymorphism by using paired DNA samples for genotyping. Cytosolic NQO1 activities differed significantly by NQO1 genotype status in whites (NQO1(whites[NQO1*1/*1])=1.3+/ 1.7 nmol/min mg versus NQO1(whites[NQO1*1/*2+NQO1*2/*2])=0.5+/-0.7 nmol/min mg, p<0.01), but not in blacks (NQO1(blacks[NQO1*1/*1])=2.6+/-3.4 nmol/min mg versus NQO1(blacks[NQO1*1/*2])=1.1+/-1.2 nmol/min mg, p=0.134). Our findings pinpoint the presence of significant interethnic differences in polymorphic hepatic NQO1 activity. PMID- 16478653 TI - Two Japanese families with hyperekplexia who have a Arg271Gln mutation in the glycine receptor alpha 1 subunit gene. AB - We report two Japanese patients from two families with hyperekplexia who have a Arg271Gln mutation in the glycine receptor alpha 1 subunit gene. The clinical course of both patients was typical for hyperekplexia, characterized by neonatal hypertonia and exaggerated startle response, and which improved gradually with age. One was associated with umbilical hernia and hip dislocation, diagnosed at 11 months, while the other was diagnosed at 1 month. Both showed positive head retraction reflex. Four Japanese families have been reported as having hyperekplexia including our cases, of which three have shown the same missense Arg271Gln mutation, most frequently found in patients from Northern Europe and the United States. PMID- 16478654 TI - Acute respiratory distress syndrome in children with severe motor and intellectual disabilities. PMID- 16478655 TI - Lymphadenectomy for papillary thyroid cancer: changes in practice over four decades. AB - AIMS: Lymphadenectomy in the management of papillary thyroid cancer (PTC) has evolved. The aim of this study was to examine the changing role of neck dissection as reflected in the practice of a large thyroid unit over four decades. METHODS: A retrospective cohort study of patients that underwent primary thyroid surgery for papillary cancer in a single unit in the period 1958-2002. Nine 5-year periods were considered and the data relevant to the treatment of the regional lymph nodes reviewed. RESULTS: Nine hundred patients with PTC underwent surgery between 1958 and 2002 of whom 32.7% underwent lymph node dissection (LND). The use of lymphadenectomy increased from 21.4% in 1958-1962 to 48.1% in 1998-2002 of which 84% underwent a selective lymph node dissection (SLND)-a dissection where the LND is determined by the extent of the disease encountered. The mean number of nodes removed during SLND was 12.6 (range 1-56) of which a mean of 3.1 (24.8%) (0-19) were involved by the disease. Cervical levels 6 and level 4 were those most frequently dissected. There was no statistically significant difference in the complication rates in patients undergoing neck dissection and those not. CONCLUSION: The four decade experience reflects a move away from modified radical neck dissection and cherry picking towards SLND. Growing evidence suggests that lymphadenopathy in adult PTC is an adverse prognostic factor. SLND, a lymphadenectomy tailored to the extent of the disease process, is the coherent treatment for PTC since it serves the dual purpose of staging as well as control of local disease. This can be achieved with little morbidity when performed in a specialist centre. PMID- 16478656 TI - Correlation between copy number of mitochondrial DNA and clinico-pathologic parameters of hepatocellular carcinoma. AB - AIMS: In the current study, we investigated possible correlations of the mtDNA copy number in hepatocellular carcinoma (HCC) with the pathological findings and prognosis. METHODS: We studied 31 HCC specimens using quantitative real-time polymerase chain reaction analysis, and the correlation between the mtDNA copy number and the clinicopathologic parameters and mutations in the D-loop region of the mitochondrial genome. RESULTS: The mtDNA copy number was reduced in HCCs compared with the corresponding non-cancerous liver tissues (p=0.002), and significantly correlated with tumour size (p=0.014) and cirrhosis (p=0.048). Patients with a low mtDNA copy number tended to show shorter 5-year survival rates than patients with a high mtDNA copy number when assessed by Kaplan-Meier curves, but not a significant (overall survival rate, 63 vs 83%; p=0.19). The copy number of HCC with mtDNA D-loop mutation or deletion was lower, but not significantly so (p=0.656, p=0.590, respectively). CONCLUSIONS: Our results indicated that a reduced copy number of mtDNA is correlated with HCC and associated with malignant potential. PMID- 16478657 TI - Cytotoxic agents in the treatment of laryngeal sarcoidosis: a case report and review of the literature. AB - SUMMARY: Sarcoidosis is a multisystem disease with various clinical manifestations. It is characterized primarily on a histopathologic basis by the presence of noncaseating granulomata. Laryngeal involvement reportedly occurs in 3-5% of cases, and it is typically localized to the supraglottic region. Patients often present with hoarseness, dysphagia, stridor, or dyspnea. Laryngoscopy typically demonstrates a pale, edematous epiglottis studded with nodules. Tissue biopsy reveals the classic noncaseating granuloma; however, the finding is not diagnostic. Sarcoidosis remains a diagnosis of exclusion to be entertained once other verifiable etiologies for granulomatous disease of the larynx, such as TB, syphilis, fungal infection, berylliosis, or Wegener's granulomatosis, have been ruled out. Systemic corticosteroids remain the mainstay of treatment; however, new steroid-sparing therapies that target the inflammatory response of sarcoidosis are currently being investigated. The case history of a patient with laryngeal sarcoidosis who was managed with the immunosuppressant azathioprine (Imuran) is summarized along with a discussion of other treatment options. PMID- 16478658 TI - Predicting mutational change in the speaking voice of boys. AB - SUMMARY: The authors investigated whether acoustic speaking voice analyses can be used to predict the beginning of mutation in 21 male members of a professional boys' choir. Over a period of 3 years before mutation, children were examined every 3 months by ear, nose, and throat (ENT) and phoniatric specialists. At the same time, the voice was evaluated acoustically using analysis features of the Goettingen Hoarseness Diagram (GHD). Irregularity component and noise component, jitter, shimmer, mean waveform correlation coefficient, and fundamental frequency were determined from recordings of the speaking voice. Significant changes of acoustic features appeared 7 and 5 months before mutation onset, which indicates that vocal function is already restricted 6 months before mutation onset. This acoustic voice analysis is therefore suitable to support the care of the professional singing voice. PMID- 16478659 TI - Videostroboscopic characteristics of young adult female smokers vs. nonsmokers. AB - OBJECTIVES: The purpose of this study was to provide a foundation of information concerning the early effects of smoking on the laryngeal structures and phonatory physiology of young adult women using videostroboscopy. STUDY DESIGN: Cross sectional, observational. METHODS: Subjects were 34 female nonsmokers and 30 female smokers (Total N=64) between the ages of 18 and 30 years of age. For the purposes of this study, a smoker was defined as any subject who, at the time of this study, had smoked at least two cigarettes per day for at least 1 year. All subjects in the nonsmoker category were those who, at the time of this study, did not smoke and who had not smoked for at least 5 years before the study. Videostroboscopic evaluations were rated by experienced judges on various parameters describing the structure and function of the laryngeal mechanism during comfortable pitch and loudness phonations. RESULTS: Results showed significant relationships between smoking and the increased presence of edema, erythema, and abnormal laryngeal color. In addition, smokers were observed to have significantly more occurrences of abnormal phase symmetry and increased glottal gap size during the closed phase of phonation. Glottal gap size was significantly correlated with the number of cigarettes smoked in the lifetime. CONCLUSIONS: The findings of increased erythema and edema in this study are consistent with previous descriptions of the effects of smoking on laryngeal structures, whereas our observations regarding glottal gap size and phase symmetry are unique contributions derived from the stroboscopic method. The results of this study indicate that observable signs of laryngeal irritation and disturbed phonatory physiology occur even in young adults with relatively brief smoking habits. PMID- 16478660 TI - Comparison of two inverse filtering methods in parameterization of the glottal closing phase characteristics in different phonation types. AB - SUMMARY: Inverse filtering (IF) is a common method used to estimate the source of voiced speech, the glottal flow. This investigation aims to compare two IF methods: one manual and the other semiautomatic. Glottal flows were estimated from speech pressure waveforms of six female and seven male subjects producing sustained vole /a/ in breathy, normal, and pressed phonation. The closing phase characteristics of the glottal pulse were parameterized using two time-based parameters: the closing quotient (C1Q) and the normalized amplitude quotient (NAQ). The information given by these two parameters indicates a strong correlation between the two IF methods. The results are encouraging in showing that the parameterization of the voice source in different speech sounds can be performed independently of the technique used for inverse filtering. PMID- 16478661 TI - A bio-basis function neural network for protein peptide cleavage activity characterisation. AB - This paper presents a novel neural learning algorithm for analysing protein peptides which comprise amino acids as non-numerical attributes. The algorithm is derived from the radial basis function neural networks (RBFNNs) and is referred to as a bio-basis function neural network (BBFNN). The basic principle is to replace the radial basis function used by RBFNNs with a bio-basis function. Each basis in BBFNN is supported by a peptide. The bases collectively form a feature space, in which each basis represents a feature dimension. A linear classifier is constructed in the feature space for characterising a protein peptide in terms of functional status. The theoretical basis of BBFNN is that peptides, which perform the same function will have similar compositions of amino acids. Because of this, the similarity between peptides can have statistical significance for modelling while the proposed bio-basis function can well code this information from data. The application to two real cases shows that BBFNN outperformed multi-layer perceptrons and support vector machines. PMID- 16478662 TI - EGFR signaling leads to downregulation of PTP-LAR via TACE-mediated proteolytic processing. AB - Proteolytic processing and ectodomain shedding have been described for a broad spectrum of transmembrane proteins under both normal and pathophysiological conditions and has been suggested as one mechanism to regulate a protein's function. It has also been documented for the receptor-like protein tyrosine phosphatase PTP-LAR, induced by treating cells with the tumor promoter TPA or the calcium ionophor A23187. Here we identified the epidermal growth factor receptor (EGFR) as both an association partner of PTP-LAR, that mediates phosphorylation of the latter, as well as an inducer of LAR-cleavage. Both overexpression of this kinase and stimulation of endogenous EGFR in various tumor cell lines were shown to induce proteolytic processing of the catalytic LAR-P-subunit. In contrast to TPA-induced shedding of PTP-LAR, EGFR-mediated cleavage did not require PKC activity. For both stimuli, however, processing of the P-subunit turned out to be dependent on the activation of the MAP kinases ERK1 and ERK2, and was completely abrogated upon pre-treating cells with Batimastat, indicating the involvement of a metalloproteinase in this pathway. Being strongly impaired in fibroblasts derived from ADAM-17/TACE-knockout-mice or tumor cells that express a dominant negative mutant of ADAM-17/TACE, cleavage of PTP-LAR is suggested to be mediated by this metalloproteinase. Paralleled by rapid reduction of cell surface localized LAR-E-subunit, EGFR-induced cleavage could be shown to lead to degradation of the catalytic LAR-P-subunit, thereby resulting in a significantly reduced overall cellular phosphatase activity of PTP-LAR. These results for the first time identify a protein tyrosine phosphatase as a potential substrate of TACE and describe proteolytic processing of PTP-LAR as a means of regulating phosphatase activity downstream and thus under the control of EGFR-mediated signaling pathways. PMID- 16478663 TI - Gradients in the self-organization of the mitotic spindle. AB - Recent evidence points at a role of protein interaction gradients around chromatin in mitotic spindle morphogenesis in large eukaryotic cells. Here, we explain how gradients can arise over distances of tens of microns around supramolecular structures from mixtures of soluble molecules. We discuss how coupled sets of such reaction diffusion processes generate the spatial information that determines the local dynamics of microtubules required to form a bipolar spindle. We argue that such reaction diffusion processes are involved in the self-organization of supramolecular structures in the cell. PMID- 16478665 TI - Diffusion tensor imaging in primary brain tumors: reproducible quantitative analysis of corpus callosum infiltration and contralateral involvement using a probabilistic mixture model. AB - Diffusion tensor imaging (DTI) has been advocated as a promising tool for delineation of the extent of tumor infiltration by primary brain tumors. First reports show conflicting results mainly due to difficulties in reproducible determination of DTI-derived parameters. A novel method based on probabilistic voxel classification for a user-independent analysis of DTI-derived parameters is presented and tested in healthy controls and patients with primary brain tumors. The proposed quantification method proved to be highly reproducible both in healthy controls and patients. Fiber integrity in the corpus callosum (CC) was measured using this quantification method, and the profiles of fractional anisotropy (FA) provided additional information of the possible extent of infiltration of primary brain tumors when compared to conventional imaging. This yielded additional information on the nature of ambiguous contralateral lesions in patients with primary brain tumors. The results show that DTI-derived parameters can be determined reproducibly and may have a strong impact on evaluation of contralateral extent of primary brain tumors. PMID- 16478664 TI - Effects of gap junction blockers on human neocortical synchronization. AB - Field potentials and intracellular recordings were obtained from human neocortical slices to study the role of gap junctions (GJ) in neuronal network synchronization. First, we examined the effects of GJ blockers (i.e., carbenoxolone, octanol, quinine, and quinidine) on the spontaneous synchronous events (duration = 0.2-1.1 s; intervals of occurrence = 3-27 s) generated by neocortical slices obtained from temporal lobe epileptic patients during application of 4-aminopyridine (4AP, 50 muM) and glutamatergic receptor antagonists. The synchronicity of these potentials (recorded at distances up to 5 mm) was decreased by GJ blockers within 20 min of application, while prolonged GJ blockers treatment at higher doses made them disappear with different time courses. Second, we found that slices from patients with focal cortical dysplasia (FCD) could generate in normal medium spontaneous synchronous discharges (duration = 0.4-8 s; intervals of occurrence = 0.5-90 s) that were (i) abolished by NMDA receptor antagonists and (ii) slowed down by carbenoxolone. Finally, octanol or carbenoxolone blocked 4AP-induced ictal-like discharges (duration = up to 35 s) in FCD slices. These data indicate that GJ play a role in synchronizing human neocortical networks and may implement epileptiform activity in FCD. PMID- 16478666 TI - Non-invasive detection of fluorescence from exogenous chromophores in the adult human brain. AB - This is the first report on results proving that fluorescence of exogenous dyes inside the human brain can be excited and detected non-invasively at the surface of the adult head. Boli of indocyanine green (ICG) were intravenously applied to healthy volunteers, and the passage of the contrast agent in the brain was monitored by detecting the corresponding fluorescence signal following pulsed laser excitation at 780 nm. Our hypothesis that the observed fluorescence signal contains a considerable cortical fraction was corroborated by performing measurements with picosecond temporal resolution and analyzing distributions of times of arrival of photons, hence taking advantage of the well-known depth selectivity of that method. Our experimental findings are explained by Monte Carlo simulations modeling the head as a layered medium and taking into account realistic bolus kinetics within the extra- and intracerebral compartment. Although a particular non-specific dye (ICG) was used, the results clearly demonstrate that fluorescence-mediated imaging of the adult human brain is generally feasible. In particular, we will discuss how these results serve as proof of concept for non-invasive fluorescence brain imaging and may thus open the door towards optical molecular imaging of the human brain. PMID- 16478667 TI - Cortical mechanisms of person representation: recognition of famous and personally familiar names. AB - Personally familiar people are likely to be represented more richly in episodic, emotional, and behavioral contexts than famous people, who are usually represented predominantly in semantic context. To reveal cortical mechanisms supporting this differential person representation, we compared cortical activation during name recognition tasks between personally familiar and famous names, using an event-related functional magnetic resonance imaging (fMRI). Normal subjects performed familiar- or unfamiliar-name detection tasks during visual presentation of personally familiar (Personal), famous (Famous), and unfamiliar (Unfamiliar) names. The bilateral temporal poles and anterolateral temporal cortices, as well as the left temporoparietal junction, were activated in the contrasts Personal-Unfamiliar and Famous-Unfamiliar to a similar extent. The bilateral occipitotemporoparietal junctions, precuneus, and posterior cingulate cortex showed activation in the contrasts Personal-Unfamiliar and Personal-Famous. Together with previous findings, differential activation in the occipitotemporoparietal junction, precuneus, and posterior cingulate cortex between personally familiar and famous names is considered to reflect differential person representation. The similar extent of activation for personally familiar and famous names in the temporal pole and anterolateral temporal cortex is consistent with the associative role of the anterior temporal cortex in person identification, which has been conceptualized as a person identity node in many models of person identification. The left temporoparietal junction was considered to process familiar written names. The results illustrated the neural correlates of the person representation as a network of discrete regions in the bilateral posterior cortices, with the anterior temporal cortices having a unique associative role. PMID- 16478668 TI - Viscoelastic properties of zonal articular chondrocytes measured by atomic force microscopy. AB - OBJECTIVE: Articular chondrocytes respond to chemical and mechanical signals depending on their zone of origin with respect to distance from the tissue surface. However, little is known of the zonal variations in cellular mechanical properties in cartilage. The goal of this study was to determine the zonal variations in the elastic and viscoelastic properties of porcine chondrocytes using atomic force microscopy (AFM), and to validate this method against micropipette aspiration. METHODS: A theoretical solution for stress relaxation of a viscoelastic, incompressible, isotropic surface indented with a hard, spherical indenter (5 microm diameter) was derived and fit to experimental stress relaxation data for AFM indentation of chondrocytes isolated from the superficial or middle/deep zones of cartilage. RESULTS: The instantaneous moduli of chondrocytes were 0.55+/-0.23 kPa for superficial cells (S) and 0.29+/-0.14 kPa for middle/deep cells (M/D) (P<0.0001), and the relaxed moduli were 0.31+/-0.15 kPa (S) and 0.17+/-0.09 kPa (M/D) (P<0.0001). The apparent viscosities were 1.15+/-0.66 kPas (S) and 0.61+/-0.69 kPa-s (M/D) (P<0.0001). Results from the micropipette aspiration test showed similar cell moduli but higher apparent viscosities, indicating that mechanical properties measured by these two techniques are similar. CONCLUSION: Our findings suggest that chondrocyte biomechanical properties differ significantly with the zone of origin, consistent with previous studies showing zonal differences in chondrocyte biosynthetic activity and gene expression. Given the versatility and dynamic testing capabilities of AFM, the ability to conduct stress-relaxation measurements using this technique may provide further insight into the viscoelastic properties of isolated cells. PMID- 16478669 TI - Biomechanical characterization and in vitro mechanical injury of elderly human femoral head cartilage: comparison to adult bovine humeral head cartilage. AB - OBJECTIVES: In vitro mechanical injury of articular cartilage is useful to identify events associated with development of post-traumatic osteoarthritis (OA). To date, many in vitro injury models have used animal cartilage despite the greater clinical relevance of human cartilage. We aimed to characterize a new in vitro injury model using elderly human femoral head cartilage and compare its behavior to that of an existing model with adult bovine humeral head cartilage. DESIGN: Mechanical properties of human and bovine cartilage disks were characterized by elastic modulus and hydraulic permeability in radially confined axial compression, and by Young's modulus, Poisson's ratio, and direction dependent radial strain in unconfined compression. Biochemical composition was assessed in terms of tissue water, solid, and glycosaminoglycan (GAG) contents. Responses to mechanical injury were assessed by observation of macroscopic superficial tissue cracks and histological measurements of cell viability following single injurious ramp loads at 7 or 70%/s strain rate to 3 or 14 MPa peak stress. RESULTS: Confined compression moduli and Young's moduli were greater in elderly human femoral cartilage vs adult bovine humeral cartilage whereas hydraulic permeability was less. Radial deformations of axially compressed explant disks were more anisotropic (direction-dependent) for the human cartilage. In both cartilage sources, tissue cracking and associated cell death during injurious loading was common for 14 MPa peak stress at both strain rates. CONCLUSION: Despite differences in mechanical properties, acute damage induced by injurious loading was similar in both elderly human femoral cartilage and adult bovine humeral cartilage, supporting the clinical relevance of animal-based cartilage injury models. However, inherent structural differences such as cell density may influence subsequent cell-mediated responses to injurious loading and affect the development of OA. PMID- 16478670 TI - Strategies for predicting and treating access induced ischemic steal syndrome. AB - Access induced ischemia is an uncommon but devastating complication for patients maintained on hemodialysis. A number of clinical risk factors have been identified to select patients at risk. Intraoperative measurement of the digital brachial index may further distinguish at-risk patients when the DBI is <0.45. Once clinically significant steal has developed, surgical strategies to treat this problem should ideally reverse the ischemia while maintaining uninterrupted access for hemodialysis. To date, the distal revascularization-interval ligation or DRIL procedure has been the most consistently successful tactic in achieving these dual objectives. A number of alternative strategies have recently been proposed and will be discussed. PMID- 16478671 TI - Prophylactic endovascular repair of small abdominal aortic aneurysm. PMID- 16478672 TI - The safety and efficacy of angioseal in therapeutic endovascular interventions. AB - OBJECTIVE: The aim of this study was to evaluate the safety and efficacy of the closure device Angioseal. MATERIAL AND METHODS: All consecutive patients, who underwent a therapeutic radiological intervention using the femoral artery approach from January 2001 to January 2005 in the Service of Vascular Surgery, Henri Mondor Hospital, Creteil, France, were prospectively included in the study. The efficacy of Angioseal was defined by the ability of the device to cover the puncture site and stop bleeding. The safety was defined by the rate of complications. RESULTS: A total of 79 Angioseal devices were employed on 77 patients. There were 62 (78.5%) males and 17 (21.5%) females. The mean age of the patients was 65.2+/-11.6 years (mean+/-SD). All Angioseal devices were deployed successfully. There were 62 (78.4%) 6F and 17 (21.6%) 8F sheaths employed during the procedures. There were two minor (2.5%) hematomas, one (1.2%) major hematoma and one (1.2%) pseudoaneurysm after the procedure. The mean time of discharge from the hospital was 2.1+/-1.8 (mean+/-SD) days. The patients were followed up for a mean of 9.0+/-9.3 (1-60 months) months. CONCLUSIONS: Angioseal provides a safe and effective way of closing the femoral artery puncture site with acceptable morbidity rates. PMID- 16478673 TI - Surgical treatment for congenital arteriovenous malformation: 10 years' experience. AB - OBJECTIVES: We report our 10 years experience of the surgical treatment of congenital arteriovenous malformation (AVM). METHODS: We retrospectively reviewed the medical records of 145 patients with AVM who visited Samsung Medical Center in Korea from 1994 to 2003. Among the 145 patients, 21 patients were operated on. Preoperative embolo/sclerotherapy was done in 20 out of the 21 patients. RESULTS: The surgically treated AVMs were 13 cases of head and neck lesions, four cases of upper extremity lesions, one case each of back lesion, uterus lesion, lower extremity lesion and multiple site lesions. There were 10 patients with the extratruncular infiltrating type, nine patients with the extratruncular limited type, one patient with a truncular superficial AV fistula and one patient with a mixed type. Fourteen cases were operated on for cosmetic reasons and since they had localized lesions, and five cases were operated on for tissue necrosis. Fourteen cases were cured by a single operation, yet seven cases needed several sessions of operation to cure the AVM or to promote wound healing after surgery. CONCLUSION: The surgical treatment of AVM is a challenging issue for vascular surgeons. To minimise the complications related to surgery, a multidisciplinary team approach should be considered. PMID- 16478674 TI - Free radicals, lipid peroxidation and the antioxidant system in the blood of cows and newborn calves around calving. AB - The oxidative stress of birth in cattle (Bos taurus) was evaluated by measuring steady state concentration of free radicals in whole blood, rate of lipid peroxidation and activity of antioxidant enzymes in erythrocytes, antioxidant capacity of blood plasma in 14 calves at birth and four times after birth until 3 weeks of age and also in their mothers at calving. The same parameters were also measured in 58 dairy cows before calving, at parturition and after calving. Free radical concentration in the blood of newborn calves was higher than in cows confirming that birth means oxidative stress for calves. Red blood cell malondialdehyde in calves was the highest at birth and following the first solid feed intake at the third week. Superoxide dismutase activity increased in calves during the first three weeks of life. Ferric reducing ability of plasma was higher in calves at birth than in cows and decreased thereafter. Higher superoxide dismutase activity in red blood cells and lower ferric reducing ability of plasma in dairy cows was found at calving compared to the average of all pre- and post-calving results. We conclude that the blood of newborn calves is well prepared to deal with the oxidative stress of birth, and that such a stress is present even when some fingerprint markers of redox imbalance show no apparent alterations. Stress of calving has minor effects on the antioxidant system of cows. PMID- 16478675 TI - Knowledge barriers to PACS adoption and implementation in hospitals. AB - PURPOSE: Drawing on the classical theory of diffusion of innovations advanced by Rogers [E.M. Rogers, Diffusion of Innovations, 4th ed., Free Press, New York, NY, 1995] and on the theory of barriers to innovation [P. Attewell, Technology diffusion and organizational learning: the case of business computing. Organ. Sci. 3 (1992) 1-19; H. Tanriverdi, C.S. Iacono, Knowledge barriers to diffusion of telemedicine. Proceedings of the 20th International Conference on Information Systems, Charlotte, NC, 1999, pp. 39-50; S. Nambisan, Y.-M. Wang, Roadblocks to web technology adoption? Commun. ACM, 42 (1) (1999) 98-101], this study seeks a better understanding of challenges faced in PACS implementations in hospitals and of the strategies required to ensure their success. METHODS: To attain this objective, we describe and analyze the process used to adopt and implement PACS at two Canadian hospitals. RESULTS: Our findings clearly demonstrate the importance of treating any PACS deployment not simply as a rollout of new technology but as a project that will transform the organization. Proponents of these projects must not lose sight of the fact that, even if technological complexity represents a significant issue, it must not garner all the project team's attention. This situation is even more dangerous, inasmuch as the greatest risk to the implementation often lies elsewhere. It would also appear to be crucial to anticipate and address organizational and behavioral challenges from the very first phase of the innovation process, in order to ensure that all participants will be committed to the project. CONCLUSIONS: In order to maximize the likelihood of PACS success, it appears crucial to adopt a proactive implementation strategy, one that takes into consideration all the technical, economic, organizational, and human factors, and does so from the first phase of the innovation process. PMID- 16478676 TI - A discriminant score based on four routine laboratory blood tests for accurate diagnosis of severe fibrosis and/or liver cirrhosis in Egyptian patients with chronic hepatitis C. AB - Liver biopsy is still recommended in most patients with chronic hepatitis C (CHC). Due to its limitations and risks, the use of non-invasive blood biomarkers has been suggested for predicting liver cirrhosis in these patients. Here, we analyzed a panel of routine blood biochemical and hematological markers of 455 Egyptians (272 males and 183 females aged 26-67 years; mean age of 47.25 years) with clinically confirmed CHC. The multivariate discriminant analysis (MDA) selected a function based on absolute values of the four routine biomarkers; score=[albumin (g/L)x0.3+platelet count (10(9)/L)x0.05]-[alkaline phosphatase (IU/L)x0.014+AST/ALT ratiox6+14]. The MDA function correctly classified 98% of the cirrhotic patients at a discriminant cut-off score=0 (i.e. less than 0 indicated liver cirrhosis and greater than 0 indicated CHC without cirrhosis) with high degrees of specificity (97%), positive predictive value (99%) and negative predictive value (92%). The MDA of the absolute values of a combination of four routine tests can efficiently indicate liver cirrhosis in CHC patients. Based on individual patient MDA score value, each patient can be simply and efficiently classified into a cirrhotic or a non-cirrhotic liver patient. PMID- 16478677 TI - Biosynthesis of novel carotenoid families based on unnatural carbon backbones: a model for diversification of natural product pathways. AB - We show that the C40 carotenoid desaturase CrtI from Pantoea ananatis (Erwinia uredovora) is capable of desaturating unnaturally long C45 and C50 carotenoid backbones in recombinant E. coli. Desaturation step number in these pathways is not very specific, and at least ten new C45 and C50 carotenoids were synthesized. We also present evidence for a novel asymmetric C40 backbone formed by the condensation of farnesyl diphosphate (C15PP) with farnesylgeranyl diphosphate (C25PP), and the subsequent desaturation of this backbone by CrtI in an atypical manner. Under some conditions, the C40, C45, and C50 carotenoid backbones synthesized in E. coli were monohydroxylated; their desaturation by CrtI in vitro led to yet more novel carotenoids. Challenging CrtI with larger-than-natural substrates in vivo has allowed us to show that this enzyme regulates desaturation step number by sensing the end groups of its substrate. Analysis of the mechanisms by which chemical diversity is generated and propagated through the nascent pathways provides insight into how natural product diversification occurs in nature. PMID- 16478678 TI - Apolipoprotein CI causes hypertriglyceridemia independent of the very-low-density lipoprotein receptor and apolipoprotein CIII in mice. AB - We have recently shown that the predominant hypertriglyceridemia in human apolipoprotein C1 (APOC1) transgenic mice is mainly explained by apoCI-mediated inhibition of the lipoprotein lipase (LPL)-dependent triglyceride (TG)-hydrolysis pathway. Since the very-low-density lipoprotein receptor (VLDLr) and apoCIII are potent modifiers of LPL activity, our current aim was to study whether the lipolysis-inhibiting action of apoCI would be dependent on the presence of the VLDLr and apoCIII in vivo. Hereto, we employed liver-specific expression of human apoCI by using a novel recombinant adenovirus (AdAPOC1). In wild-type mice, moderate apoCI expression leading to plasma human apoCI levels of 12-33 mg/dl dose-dependently and specifically increased plasma TG (up to 6.6-fold, P < 0.001), yielding the same hypertriglyceridemic phenotype as observed in human APOC1 transgenic mice. AdAPOC1 still increased plasma TG in vldlr(-/-) mice (4.1 fold, P < 0.001) and in apoc3(-/-) mice (6.8-fold, P < 0.001) that were also deficient for the low-density lipoprotein receptor (LDLr) and LDLr-related protein (LRP) or apoE, respectively. Thus, irrespective of receptor-mediated remnant clearance by the liver, liver-specific expression of human apoCI causes hypertriglyceridemia in the absence of the VLDLr and apoCIII. We conclude that apoCI is a powerful and direct inhibitor of LPL activity independent of the VLDLr and apoCIII. PMID- 16478680 TI - Phenotypic discordance in three siblings affected by atypical cystic fibrosis with the F508del/D614G genotype. AB - We report an example of atypical CF, i.e., a family in which three siblings were affected by late-diagnosed mild CF, and showed discordant pulmonary and pancreatic phenotypes. Sibling no. 1 (male), showed a severe pulmonary involvement and pancreatic sufficiency; sibling no. 2 (female) showed a mild pulmonary disease with pancreatic sufficiency; sibling no. 3 (male) had a very mild pulmonary expression and pancreatic insufficiency. The sweat test was altered in all three siblings, and all had intestinal occlusion in young age. The whole scanning of CFTR revealed the rare F508del/D614G genotype. The discordance of clinical expression within the same family reinforces the putative role of modifier genes of CF phenotype. PMID- 16478679 TI - Intestinal levels of anandamide and oleoylethanolamide in food-deprived rats are regulated through their precursors. AB - The anorectic lipid oleoylethanolamide and the orexigenic lipid anandamide both belong to the group of N-acylethanolamines that are generated by the enzyme N acylphosphatidylethanolamine-hydrolyzing phospholipase D. The levels of the two bioactive lipids were investigated in rat intestines after 24 h of starvation as well as after 1 and 4 h of re-feeding. Total levels of precursor phospholipids and N-acylethanolamines were decreased upon food-deprivation whereas the level of the anandamide precursor molecule was significantly increased. The level of 2 arachidonoyl-glycerol was unchanged as was the activity of N-acyltransferase, N acylphosphatidylethanolamine-hydrolyzing phospholipase D, and fatty acid amide hydrolase upon starvation and re-feeding. It is concluded that remodeling of the amide-linked fatty acids of N-acylphosphatidylethanolamine is responsible for the opposite effects on levels of anandamide and oleoylethanolamide in intestines of food-deprived rats and not an alternative biochemical route for anandamide synthesis. Furthermore, linoleoylethanolamide, which accounted for more than 50 mol% of the endogenous pool of N-acylethanolamines, was found not to have the same inhibitory effect on food intake, as did oleoylethanolamide following oral administration. PMID- 16478681 TI - Understanding high pressure stability of helical conformation of oligopeptides and helix bundle protein high pressure FT-IR and RISM theoretical studies. AB - The pressure effect on conformational equilibria of simple organic compounds and the pressure denaturation of proteins have been well investigated by using vibrational spectroscopy. However, there was no systematic investigation of the pressure effect on conformational equilibria of oligopeptides, which are located between the simple organic compounds and proteins. Here, we review the recent vibrational spectroscopic and theoretical studies of the pressure effect on conformational equilibria of model oligopeptides and helix bundle protein in aqueous solution. PMID- 16478683 TI - High pressure macromolecular crystallography: the 140-MPa crystal structure at 2.3 A resolution of urate oxidase, a 135-kDa tetrameric assembly. AB - We report the three-dimensional structure determined by high-pressure macromolecular crystallography (HPMX) of a 135-kDa homo-tetrameric enzyme, urate oxidase from Aspergillus flavus complexed with its potent inhibitor 8-azaxanthin. Urate oxidase crystals are quite sensitive to pressure, as three-dimensional order is lost at about 180 MPa. A highly complete 2.3 A resolution data set was collected at 140 MPa, close to the critical pressure. Crystal structures at atmospheric pressure and at high pressure were refined in the orthorhombic space group I222 with final crystallographic R factors 14.1% and 16.1%, respectively. The effect of pressure on temperature factors, ordered water molecules, hydrogen bond lengths, contacts, buried surface areas as well as cavity volume was investigated. Results suggest that the onset of disruption of the tetrameric assembly by pressure has been captured in the crystalline state. PMID- 16478684 TI - Pharmacogenomics: an in-house advantage? PMID- 16478682 TI - Synergetic effects of pressure and chemical denaturant on protein unfolding: stability of a serine-type carboxyl protease, kumamolisin. AB - Kumamolisin, a serine carboxyl proteinase, is very stable and hardly denatured by single perturbation of a chemical denaturant (urea), pressure (<500 MPa) or temperature (<65 degrees C). In order to investigate the cooperative effects of these three denaturing agents, DSC, CD, intrinsic fluorescence, and fourth derivative UV absorbance were measured under various conditions. By application of pressure to kumamolisin in 8 M urea solution, substantial red-shift in the center of fluorescence emission spectral mass was observed, and the corresponding blue-shift was observed for two major peaks in fourth derivative UV absorbance, under the similar urea-containing conditions. The denaturation curves were analyzed on the basis of a simple two-state model in order to obtain thermodynamic parameters (DeltaV, DeltaG, and m values), and the combined effects of denaturing agents are discussed, with the special interest in the large cavity and neighboring Trp residue in kumamolisin. PMID- 16478685 TI - Using epidemiology and archaeology to unearth new drug targets for rheumatoid arthritis therapy. AB - Epidemiological and archaeological evidence suggests that RA could be a consequence of enhanced immunity to Mycobacterium tuberculosis, and that by understanding this connection, new RA drug targets may be uncovered. PMID- 16478686 TI - The role of neuroinflammation in neuropathic pain: mechanisms and therapeutic targets. AB - Neuroinflammation is a proinflammatory cytokine-mediated process that can be provoked by systemic tissue injury but it is most often associated with direct injury to the nervous system. It involves neural-immune interactions that activate immune cells, glial cells and neurons and can lead to the debilitating pain state known as neuropathic pain. It occurs most commonly with injury to peripheral nerves and involves axonal injury with Wallerian degeneration mediated by hematogenous macrophages. Therapy is problematic but new trials with anti cytokine agents, cytokine receptor antibodies, cytokine-signaling inhibitors, and glial and neuron stabilizers provide hope for future success in treating neuropathic pain. PMID- 16478687 TI - New drug candidates and therapeutic targets for tuberculosis therapy. AB - Despite advances in chemotherapy and the BCG (Bacillus Calmette-Guerin) vaccine, tuberculosis remains a significant infectious disease. Although it can be cured, the therapy takes at least 6-9 months, and the laborious and lengthy treatment brings with it dangers of noncompliance, significant toxicity and drug resistance. The increasing emergence of drug resistance and the problem of mycobacterial persistence highlight the need to develop novel TB drugs that are active against drug resistant bacteria but, more importantly, kill persistent bacteria and shorten the length of treatment. Recent new and exciting developments in tuberculosis drug discovery show good promise of a possible revolution in the chemotherapy of tuberculosis. PMID- 16478688 TI - The potential impact of structural genomics on tuberculosis drug discovery. AB - Mycobacterium tuberculosis, the causative agent of tuberculosis (TB) in humans, is a devastating infectious organism that kills approximately two million people annually. The current suite of antibiotics used to treat TB faces two main difficulties: (i) the emergence of multidrug-resistant (MDR) strains of M. tuberculosis, and (ii) the persistent state of the bacterium, which is less susceptible to antibiotics and causes very long antibiotic treatment regimes. The complete genome sequences of a laboratory strain (H37Rv) and a clinical strain (CDC1551) of M. tuberculosis and the concurrent identification of all the open reading frames that encode proteins within this organism, present structural biologists with a wide array of protein targets for structure determination. Comparative genomics of the species that make up the M. tuberculosis complex has also added an array of genomic information to our understanding of these organisms. In response to this, structural genomics consortia have been established for targeting proteins from M. tuberculosis. This review looks at the progress of these major initiatives and the potential impact of large scale structure determination efforts on the development of inhibitors to many proteins. Increasing sophistication in structure-based drug design approaches, in combination with increasing numbers of protein structures and inhibitors for TB proteins, will have a significant impact on the downstream development of TB antibiotics. PMID- 16478689 TI - Mining chemical structural information from the drug literature. AB - It is easier to find too many documents on a life science topic than to find the right information inside these documents. With the application of text data mining to biological documents, it is no surprise that researchers are starting to look at applications that mine out chemical information. The mining of chemical entities--names and structures--brings with it some unique challenges, which commercial and academic efforts are beginning to address. Ultimately, life science text data mining applications need to focus on the marriage of biological and chemical information. PMID- 16478690 TI - Computational chemistry-driven decision making in lead generation. AB - Novel starting points for drug discovery projects are generally found either by screening large collections of compounds or smaller more-focused libraries. Ideally, hundreds or even thousands of actives are initially found, and these need to be reduced to a handful of promising lead series. In several sequential steps, many actives are dropped and only some are followed up. Computational chemistry tools are used in this context to predict properties, cluster hits, design focused libraries and search for close analogues to explore the potential of hit series. At the end of hit-to-lead, the project must commit to one, or preferably a few, lead series that will be refined during lead optimization and hopefully produce a drug candidate. Striving for the best possible decision is crucial because choosing the wrong series is a costly one-way street. PMID- 16478691 TI - Airborne infectious disease and the suppression of pulmonary bioaerosols. AB - The current understanding of airborne pathogen spread in relation to the new methods of suppressing exhaled bioaerosols using safe surface-active materials, such as isotonic saline, is reviewed here. We discuss the physics of bioaerosol generation in the lungs, what is currently known about the relationship between expired bioaerosols and airborne infectious disease and current methods of airborne infectious disease containment. We conclude by reviewing recent experiments that suggest the delivery of isotonic saline can significantly diminish exhaled aerosol--generated from airway lining fluid in the course of natural breathing. We also discuss these implications in relation to airborne infectious disease control. PMID- 16478692 TI - MRI-guided immunotherapy development for multiple sclerosis in a primate. AB - Multiple sclerosis is a serious neurological disease that affects 1 in 1000 young adults in Europe and the USA. The development of an effective therapy for this enigmatic disease is plagued by the failure of many treatments to reproduce in patients the promising effects observed in animal models. This review describes a new preclinical model in a non-human primate that might help to bridge the gap between currently used animal models and the patients. PMID- 16478693 TI - Harnessing in vivo siRNA delivery for drug discovery and therapeutic development. AB - The use of RNA interference (RNAi) is spreading rapidly to nearly every aspect of biomedical research. The gene silencing capability of RNAi is being used to study individual gene's biological function and role in biochemical pathways. However, the efficacy of RNAi depends upon efficient delivery of the intermediates of RNAi, short interfering RNA (siRNA) and short hairpin RNA (shRNA) oligonucleotides. The delivery challenge is even greater when the aim is to inhibit the expression of target genes in animal models. Although i n vivo delivery of siRNA is complicated and challenging, recent results are encouraging. In this review, the latest developments of in vivo delivery of siRNA and the crucial issues related to this effort are addressed. PMID- 16478694 TI - Human glial cell culture models of inflammation in the central nervous system. AB - Research into human central nervous system (CNS) disorders has traditionally focused on interconnecting neurons, thought to be the most important functional elements in the CNS. Consequently, animal models have developed as the central paradigm in CNS drug development. However, evidence is accumulating that suggests glial cells play a much more important role in health and disease in the CNS than has been previously acknowledged. Brain development, neurotransmission, inflammatory and neuroprotective pathways and blood-brain barrier functions rely on glial cells. It is also the case that human glial cell cultures adequately mimic in vivo glial cell behaviour, providing a novel and valuable tool for CNS drug discovery and development. PMID- 16478695 TI - Elimination mechanisms of therapeutic monoclonal antibodies. AB - Targeted therapies using monoclonal antibodies have achieved important therapeutic applications in the treatment of various human diseases. Understanding the factors that impact the pharmacokinetics of monoclonal antibodies is of high importance for effective therapy. Many factors related to the target antigen, antibody and patients can affect antibody elimination. Evaluation of these factors will facilitate the understanding of the processes involved in antibody elimination. PMID- 16478697 TI - Staging of non-small cell lung cancer (NSCLC). AB - Staging of non-small lung cancer (NSCLC) uses the TNM classification and is undertaken to identify those patients who are surgical candidates, either initially or after chemo-radiotherapy, and to differentiate patients who will be treated radically from those requiring palliation and to plan radiotherapy fields. Computed tomography and magnetic resonance imaging (MRI) are used in staging and provide anatomical information but have well known limitations in differentiating reactive from malignant nodes, fibrosis from active disease and in defining the extent of invasion. MRI, with its superior soft tissue contrast provides optimal information on brachial plexus and central nervous system involvement. Functional imaging using [2-(18F)]fluorodeoxyglucose positron emission tomography is increasingly being used to provide unique information and when combined with anatomic imaging will provide better staging information for both local disease and the extent of metastases. PMID- 16478698 TI - Contrast enhanced ultrasound of breast cancer. AB - The importance of ultrasound examination in the diagnosis of breast cancer has been widely demonstrated. During the last few years, the introduction of ultrasound contrast media has been considered a promising tool for studying the vascular pattern of focal lesions within the breast. Our purpose was to assess whether contrast-enhanced (CE) ultrasound examination, performed using specific contrast imaging modes, can be helpful for detection and characterization of breast lesions, and for prediction of the response of breast cancer to therapy. PMID- 16478699 TI - [Scorpion stings: a public health problem in Morelos (Mexico)]. AB - Scorpion stings represent a major public health problem in Mexico. Their annual incidence is estimated at 150,000 cases; 800-1,000 people die from them each year, 72.5% of whom are children younger than 5 years old. The states most affected are Aguascalientes, Colima, Durango, Guanajuato, Guerrero, Jalisco, Michoacan, Morelos, Oaxaca, Puebla, Sinaloa, and Zacatecas. Morelos is an endemic zone, and scorpion stings are relatively frequent, but the indigenous population underestimates the risks. In this locality, scorpion stings lead to high morbidity, with an average of 10,219 cases each year, and a prevalence varying from 584.86 per 100,000 inhabitants in 1994 to 2043.3 per 100,000 inhabitants in 2003. Because of this upsurge, systematic medical supervision and serum therapy are necessary if scorpion poisoning is suspected in a child. The indigenous community as well as tourists visiting Mexico must be informed about the substantial risk of scorpion poisoning. PMID- 16478700 TI - [Mass antibiotic prophylaxis against cholera in the New Bell central prison in Douala during the 2004 epidemic]. AB - In early January 2004, cases of severe watery diarrhea were reported in Douala, the economic capital of Cameroon (estimated population: 2.4 million inhabitants). Three stool samples examined at the Cameroon National Reference Laboratory grew Vibrio cholerae serogroup O1, later identified by the Pasteur Institute in Paris, France, as serotype Inaba. On 19 January, the provincial health authorities declared an outbreak of cholera. The epidemic lasted until that September. In all, 5,020 cholera cases were reported, and 69 deaths from cholera among hospital patients. The overall attack rate for Douala was 209 cases per 100,000 inhabitants, with a case-fatality ratio of 1.37%. New Bell Central Prison, the sole penitentiary facility for Douala, is situated in the center of the town, near the biggest market. It was originally built in 1930 for 700 prisoners but now houses an average of roughly 3,100 inmates. Living and sanitary conditions in the prison are deplorable. Half of the cells house more than 150 inmates with a surface area of less than 0.20 m2 per inmate. Approximately 400 people--visitors, new admissions, and discharged prisoners--enter and leave the prison. In February 2004, five suspected cases of cholera were reported in the prison's hospital ward. Immediate measures were taken to prevent an explosive spread of cholera within the prison: a) suspected cases were treated with rehydration therapy, antibiotics, and isolation; and b) preventive antibiotic treatment, consisting of a single 300-mg dose of doxycycline, was administered to all 3,036 prisoners and 164 prison staff members. No significant side effects were observed. Despite a reinforced surveillance system, no new cholera cases were reported except two suspected cases in June 2004 (four months later), and their diagnoses could not be confirmed. A cholera epidemic in an urban area mandates rigorous epidemiological surveillance system and provisions for safety stocks of therapeutic and prophylactic drugs in closed at-risk settings, such as prisons. Collective single-dose prophylaxis with an antibiotic the efficacy of which is confirmed by in vitro antibiotic susceptibility testing can provide immediate protection not available by other preventive measures (education, sanitation, immunization) against a cholera epidemic in a prison. PMID- 16478701 TI - [Drug prescription practices in public medical centres in Gabon]. AB - CONTEXT OF THE PROBLEM: Inappropriate drug use constitutes a major problem of health programs in developing countries. The objective of this study was to describe and assess the quality of drug prescription practices in public medical centres in Gabon. METHOD: Using the Drug Use Prescribing Indicators developed collaboratively by the World Health Organization (WHO) and the International Network for Rational Use of Drugs (INRUD), we retrospectively evaluated the drugs prescribed during 1,200 randomly selected outpatient consultations that took place in 20 randomly selected public medical centres during the period between July 1, 1997 and June 30, 1998. RESULTS: A total of 3,054 drugs were prescribed during the 1,200 consultations under study, for an average of 2.6 (Standard Deviation: +/- 1.2) drugs per consultation. Of all the drugs prescribed, 13.5% (95% Confidence Interval (CI): 12.3-14.7) were prescribed under their international nonproprietary name (INN), while 57.1% (CI: 55.3-58.9) appeared on the national list of essential drugs. 64.5% (CI: 61.8-67.2) consultations resulted in the prescription of an antibiotic, while 21.8 % (CI: 19.5-24.1) led to the prescription of at least one injection. On average, six key drugs out of eleven were available in the medical centres visited. CONCLUSION: The results of this study suggest that drug prescriptions are far from optimal in the public medical centers in Gabon. Among other things, the prescription of drugs under their INN is very low. These findings highlight the need to develop and implement programs that seek to improve the quality of drug prescriptions in Gabon. PMID- 16478702 TI - [Community participation in health agencies in an urban health district of Bukavu (Democratic Republic of the Congo)]. AB - We sought to evaluate the level of community participation in health agencies in the Bukavu urban health district, an alternative approach to assessing primary health care that can be especially useful in combating poverty. Accordingly we conducted a transversal survey of a population including 113 members of 12 functional health committees and the chief of the Bukavu Urban Health District, a physician. Subjects were most often 35 to 65 years of age, male, salaried employees, married, and Roman Catholic, and had not completed secondary school. Means were compared with Anova (analysis of variance). The level of participation by these health committee members is poor (29.0%), and very similar regardless of the health sector: Bagira (26.5%), Ibanda (25.9%), and Kadutu (32.6%). Relevant factors include the weak commitment of district health authorities, the poor participation by committee members in performing their roles, and their low level of motivation to participate. The results showed no significant difference between the three health sectors in the motivating power of roles and attributions. The authors recommend that health district authorities commit themselves to the selection, training and supervision of health committees. PMID- 16478703 TI - [Ocular disorders causing blindness in working-age outpatients: a prospective study at the African Institute of Tropical Ophthalmology (IOTA, Bamako, Mali)]. AB - The objective of this study was to determine the different groups of ocular disorders causing low vision or blindness among working-age patients consulting at the African Institute of Tropical Ophthalmology (IOTA) in Bamako, Mali. METHODS: This prospective study took place from 1 February through 8 April, 2003, at IOTA. It included all new patients aged between 18 and 50 years, regardless of whether they worked, but excluding students. RESULTS: 828 patients were recruited: 384 men (46.4%) and 444 women (53.6%). The average age was 35.7 years. The rate of bilateral blindness was 5.8%, of low vision 8.5%, and of blindness in one eye 11.5%. Men and women did not differ significantly in rates of bilateral blindness (p = 0.77) or low vision (p = 0.24), but blindness in a single eye affected men (67.4%) more often than women (32.6%, p = 0.01). While the frequency of bilateral blindness and low vision increased with age, blindness in one eye affected all ages. We observed no significant differences according to ethnic group in the prevalence of bilateral blindness and low vision, both observed most frequently among housewives, farmers and low-income workers, most illiterate. The major causes of bilateral blindness were cataracts 29.2%, glaucoma 22.9%, trachoma 14.6%, ocular manifestations linked to HIV/AIDS 8.3%, and non glaucomatous optical neuropathies 8.3%. Half the cases (50.5%) of blindness in one eye resulted from ocular trauma, and five cases from HIV/AIDS (5.3%). Low vision resulted from impaired refraction 22.9%, cataract 17.2%, trachoma 15.7%, glaucoma 10% and corneal opacities 5.7%. DISCUSSIONS: The high rate of blindness and low vision are associated with the study site: a reference center whose population was composed completely of patients with vision problems. The causes of blindness and low vision have not changed over time except for the ocular manifestations of HIV/AIDS, which is among the most important current causes of blindness, because of the lack of antiretroviral treatments in sub-Saharan Africa. If nothing stops this pandemic, it will soon modify epidemiological data about blindness. CONCLUSION: Blindness and visual deficiencies, already a public health problem, will become an important socioeconomic burden in sub-Saharan Africa and may even stop progress among low-income countries. PMID- 16478704 TI - [Multinodular diving goiters: 100 cases in Morocco]. AB - OBJECTIVE: To assess 100 cases of multinodular diving goiters, the authors review the literature to compare the epidemiology, clinical pictures, additional required work-up, treatments, complications, and sequelae. METHOD: Records of 100 cases of multinodular diving goiters were collected in the surgical department of the Military Hospital of Marrakesh in Morocco from 1991 through 2004. They accounted for 6% of all goiters. The sex ratio was clearly female, and the mean age 50 years. The clinical symptoms of diving goiters involves mainly signs of compression, with dyspnea seen in 50% of cases. Thyroid dysfunction was found in only 25% of our patients. A diagnosis of diving goiter must be suspected when there are signs of mediastinal compression and a palpable cervical goiter, as seen in all our patients. The diagnosis can often be confirmed with thoracic radiography and thyroid scintigraphy. Treatment is mainly surgical and depends on disease course. Cervicotomy was performed in 97% of our patients and was sufficient to extract even most voluminous goiters and those deepest in the mediastinum. RESULTS: Immediate operative results were satisfactory. More long term results were also generally satisfactory, except for 4 cases of recurrent paralysis and 5 cases of hypoparathyroidism. Both have been reported by several authors. CONCLUSION: Surgical management of multinodular diving goiters is necessary. In general, cervicotomy is sufficient, and the results generally satisfactory, except some complications and neoplasms. PMID- 16478705 TI - [HIV infection and modification of social relationships: study of 188 HIV infected persons in Ouagadougou (Burkina Faso)]. AB - HIV infection creates diverse representations among sick people, their families, the community and healthcare workers, representations that modify social relationships. To analyze the interpersonal relationships in families of people living with HIV, we began a cross-sectional descriptive study in 2001 of a population of HIV patients treated at an outpatient center in Ouagadougou. Our objectives were to: 1 degree) describe the changes in the social relationships between the subjects and members of their families; 2 degrees) itemize the difficulties they met within their families and identify the different types of discrimination they may suffer there. The systematic inclusion of HIV patients at the treatment center made it possible to question 188 subjects, including 122 women and 66 men aged 19 to 55 years. With the subjects' consent, 66 family members were also interviewed. According to these patients, the principal significant events experienced after disclosing their HIV status were absence of sexual intercourse with the partner (16.0%), rejection by their family (9.6%), taunting, criticism and contempt (6.4%). This conduct came from siblings (25%), other family members (20.8%), neighbours (20.8%), in-laws (16.7%), and the spouse (4.2%). Among our subjects, 74.5% informed their families that they were HIV positive. Of the family members questioned, 82.1% considered the subject to be a victim and 17.9% considered him or her guilty. After receiving this information 21.5% of the families modified their relationships with the subjects. Despite information and awareness campaigns, HIV infection is laden with emotion and prejudice and subjects its victims to discriminatory behavior. Further public education is necessary to improve acceptance of people with this disease by their family and friends. PMID- 16478706 TI - [Psychiatric consultations in school settings: 170 cases at a school and university health center (CSSU) from 1995 through 2001]. AB - This retrospective study of 170 files from the School and University Health Center in the municipality of Plateau shows that psychiatric disorders affected students at all levels, with 72.4% of the cases involving secondary school students. Depressive disorders were most common, accounting for 49% of the cases. Almost no students (5.3%) suspended their studies during the school year. This observation contrasts with the common belief that pupils with mental disorders leave school permanently. PMID- 16478707 TI - [Child malnutrition and maternal overweight in same households in poor urban areas of Benin]. AB - The coexistence of child malnutrition and maternal overweight in the same households typifies rapid nutrition transition in developing countries (DCs). It is reportedly less common in Africa than in Latin America or Asia, but the phenomenon is still little documented. The purpose of our study in poor urban neighbourhoods of the capital city of Benin (West Africa) was to assess the magnitude of the overlap of child protein-energy malnutrition (PEM) and maternal overweight within households, and to compare these households with other nutritional phenotypes in terms of socio-economic circumstances and diet quality. Our hypothesis was that both child PEM and maternal overweight could stem from poor socio-economic conditions, including lack of sanitation, and poor diet quality. Food diversity was used as an index of diet quality, or the qualitative dimension of food security. A random sample of 148 households that included a least the biological mother, one child between 6 and 59 months of age and a second one between 5 and 11 years was selected in two poor neighbourhoods of the capital city of Cotonou to assess the prevalence of "double burden" households and of other nutritional phenotypes of households: with PEM only in at least one child; with maternal overweight only; and without PEM or overweight. Body weights and heights of mothers and of the two targeted children were measured. As long as one child had low weight-for-height or height-for-age (z-score <-2.0), the household came under the "PEM" type. In mothers, overweight was defined as body mass index (BMI) > or = 25, obesity > or = 30 and chronic energy deficiency <18.5. We retained 126 households for interviews with mothers on socio-economic circumstances and food diversity. A socio-economic status (SES) score was constructed on the basis of household amenities and maternal education. Food insecurity was based on reported shortage of food in the last year in the household. Based on the frequency of consumption of 13 different food groups in the previous week, a food diversity score was computed. Overall 35.5% of children were malnourished, and school-age children had a worse nutritional status than under-5 children: 41% and 30% PEM (chronic or acute or both), respectively. The rate of maternal overweight was 39.1% including 15.5% of obesity. Child PEM coexisted with maternal overweight or obesity in 16.2% of the households; 27.7% of households had PEM only, 23% overweight only, 20.3% showed no malnutrition or overweight, and 12.8% had an underweight mother. Maternal BMI status was significantly associated with both children's weight-for-height z-score, particularly the elder one. The rate of child malnutrition, particularly wasting, was significantly higher among underweight mothers and lower in overweight mothers . Underweight mothers were merged with mothers with BMI < or = 25 for the remaining analyses. Households with overweight mothers tended to enjoy relatively better socio-economic conditions--higher SES, higher maternal education, less food insecurity, better household sanitation; they also tended to have a more diversified diet. This is in contrast with PEM households. Dual burden households shared several socio-economic features with the PEM households, except for a higher (not significant) SES score. Dual burden households also had the lowest food diversity score of all household types. Logistical regression models revealed that a relatively higher SES level was associated with a higher likelihood of maternal overweight in PEM households, whereas poor household sanitation increased the odds of PEM among maternal overweight households. Food diversity appeared significantly associated with a lower likelihood of dual burden in all types of households. The study highlights the importance of addressing the double burden of malnutrition and overweight even in poor areas of low income countries of West Africa. It suggests that prevention efforts should be aimed at improving diet quality and sanitation in poor urban households. PMID- 16478708 TI - [Advances in the diagnosis and management of gastrointestinal tumours: 5 cases]. AB - INTRODUCTION: Gastrointestinal stromal tumours (GIST) are the most frequent mesenchymatous tumours of the digestive tract. Options for diagnosis and treatment have developed rapidly in recent years. The authors report five cases of GIST. CASES: The study concerns 5 patients: 3 men and 2 women, with a mean age at diagnosis of 39.8 years. We describe the circumstances of discovery and the clinical and morphological characteristics of these tumours. They were located in the oesophagus (1 case), stomach (2 cases), small intestine (1 case) and mesentery (1 case). Tumour size ranged from 4-20 cm. Liver metastasis was identified at initial diagnosis for one patient. Immunohistochemical analysis identified expression of CD34, CD117 and S-100 proteins in all cases and smooth muscle actin in 2 cases. All the patients underwent surgical resection for GIST, and one received chemotherapy. No patient received imatinib treatment. After a mean follow-up of 40 months: one patient presented peritoneal carcinosis at one year, and 3 patients liver metastasis at 2 years; one of the latter died. DISCUSSION: The cases reported here are original by the rarity of their localization (especially the oesophagus and the mesentery). Diagnosis and treatment options for patients are described in a discussion of the recent advances in the field. CONCLUSION: GIST are the most frequently mesenchymatous tumours of the digestive tract. They must be recognized, especially in view of their frequency. Optimal management today involves complete tumour resection and imatinib. PMID- 16478709 TI - Atomic-level simulations of seeman DNA nanostructures: the paranemic crossover in salt solution. AB - We use molecular dynamics (MD) simulations to understand the structure and stability of various paranemic crossover (PX) DNA molecules, synthesized recently by Seeman and co-workers at New York University. These studies include all atoms of the PX structures with an explicit description of solvent and ions. The average dynamics structures over the last 1 ns of the 3-ns simulation preserve the Watson-Crick hydrogen bonding as well as the helical structure. The root mean square deviation in coordinates with respect to the MD averaged structure converges to 2-3 A for PX55, PX65, and PX85, but for PX75 and PX95 the root mean square deviation in coordinates exhibits large fluctuations, indicating an intrinsic instability. The PX structures are structurally more rigid compared to the canonical B-DNA without crossover. We have developed a strain energy analysis method based on the nearest-neighbor interaction and computed the strain energy for the PX molecules compared to the B-DNA molecules of the same length and sequence. PX65 has the lowest calculated strain energy (approximately -0.77 kcal/mol/bp), and the strain increases dramatically for PX75, PX85, and PX95. PX55 has the highest strain energy (approximately 1.85 kcal/mol/bp) making it unstable, which is in accordance with the experimental results. We find that PX65 has helical twist and other helical structural parameters close to the values for normal B-DNA of similar length and sequence. Vibrational mode analysis shows that compared to other PX motifs, PX65 has the smallest population of the low frequency modes that are dominant contributors for the conformational entropy of the PX DNA structures. All these results indicate that PX65 is structurally more stable compared to other PX motifs, in agreement with experiments. These results should aid in designing optimized DNA structures for use in nanoscale components and devices. PMID- 16478710 TI - Diels-Alder cycloadditions in water for the straightforward preparation of peptide-oligonucleotide conjugates. AB - The Diels-Alder reaction between diene-modified oligonucleotides and maleimide derivatized peptides afforded peptide-oligonucleotide conjugates with high purity and yield. Synthesis of the reagents was easily accomplished by on-column derivatization of the corresponding peptides and oligonucleotides. The cycloaddition reaction was carried out in mild conditions, in aqueous solution at 37 degrees C. The speed of the reaction was found to vary depending on the size of the reagents, but it can be completed in 8-10 h by reacting the diene oligonucleotide with a small excess of maleimide-peptide. PMID- 16478711 TI - High-throughput trapping of secretory pathway genes in mouse embryonic stem cells. AB - High-throughput gene trapping is a random approach for inducing insertional mutations across the mouse genome. This approach uses gene trap vectors that simultaneously inactivate and report the expression of the trapped gene at the insertion site, and provide a DNA tag for the rapid identification of the disrupted gene. Gene trapping has been used by both public and private institutions to produce libraries of embryonic stem (ES) cells harboring mutations in single genes. Presently, approximately 66% of the protein coding genes in the mouse genome have been disrupted by gene trap insertions. Among these, however, genes encoding signal peptides or transmembrane domains (secretory genes) are underrepresented because they are not susceptible to conventional trapping methods. Here, we describe a high-throughput gene trapping strategy that effectively targets secretory genes. We used this strategy to assemble a library of ES cells harboring mutations in 716 unique secretory genes, of which 61% were not trapped by conventional trapping, indicating that the two strategies are complementary. The trapped ES cell lines, which can be ordered from the International Gene Trap Consortium (http://www.genetrap.org), are freely available to the scientific community. PMID- 16478712 TI - On-chip non-equilibrium dissociation curves and dissociation rate constants as methods to assess specificity of oligonucleotide probes. AB - Nucleic acid hybridization serves as backbone for many high-throughput systems for detection, expression analysis, comparative genomics and re-sequencing. Specificity of hybridization between probes and intended targets is always critical. Approaches to ensure and evaluate specificity include use of mismatch probes, obtaining dissociation curves rather than single temperature hybridizations, and comparative hybridizations. In this study, we quantify effects of mismatch type and position on intensity of hybridization signals and provide a new approach based on dissociation rate constants to evaluate specificity of hybridized signals in complex target mixtures. Using an extensive set of 18mer oligonucleotide probes on an in situ synthesized biochip platform, we demonstrate that mismatches in the center of the probe are more discriminating than mismatches toward the extremities of the probe and mismatches toward the attached end are less discriminating than those toward the loose end. The observed destabilizing effect of a mismatch type agreed in general with predictions using the nearest neighbor model. Use of a new parameter, specific dissociation temperature (T(d-w), temperature of maximum specific dissociation rate constant), obtained from probe-target duplex dissociation profiles considerably improved the evaluation of specificity. These results have broad implications for hybridization data obtained from complex mixtures of nucleic acids. PMID- 16478714 TI - Genotyping pooled DNA using 100K SNP microarrays: a step towards genomewide association scans. AB - The identification of quantitative trait loci (QTLs) of small effect size that underlie complex traits poses a particular challenge for geneticists due to the large sample sizes and large numbers of genetic markers required for genomewide association scans. An efficient solution for screening purposes is to combine single nucleotide polymorphism (SNP) microarrays and DNA pooling (SNP-MaP), an approach that has been shown to be valid, reliable and accurate in deriving relative allele frequency estimates from pooled DNA for groups such as cases and controls for 10K SNP microarrays. However, in order to conduct a genomewide association study many more SNP markers are needed. To this end, we assessed the validity and reliability of the SNP-MaP method using Affymetrix GeneChip Mapping 100K Array set. Interpretable results emerged for 95% of the SNPs (nearly 110,000 SNPs). We found that SNP-MaP allele frequency estimates correlated 0.939 with allele frequencies for 97 605 SNPs that were genotyped individually in an independent population; the correlation was 0.971 for 26 SNPs that were genotyped individually for the 1028 individuals used to construct the DNA pools. We conclude that extending the SNP-MaP method to the Affymetrix GeneChip Mapping 100K Array set provides a useful screen of >100,000 SNP markers for QTL association scans. PMID- 16478713 TI - Affinity selection of DNA-binding protein complexes using mRNA display. AB - Comprehensive analysis of DNA-protein interactions is important for mapping transcriptional regulatory networks on a genome-wide level. Here we present a new application of mRNA display for in vitro selection of DNA-binding protein heterodimeric complexes. Under improved selection conditions using a TPA responsive element (TRE) as a bait DNA, known interactors c-fos and c-jun were simultaneously enriched about 100-fold from a model library (a 1:1:20 000 mixture of c-fos, c-jun and gst genes) after one round of selection. Furthermore, almost all kinds of the AP-1 family genes including c-jun, c-fos, junD, junB, atf2 and b atf were successfully selected from an mRNA display library constructed from a mouse brain poly A(+) RNA after six rounds of selection. These results indicate that the mRNA display selection system can identify a variety of DNA-binding protein complexes in a single experiment. Since almost all transcription factors form heterooligomeric complexes to bind with their target DNA, this method should be most useful to search for DNA-binding transcription factor complexes. PMID- 16478715 TI - Nucleic acid visualization with UCSF Chimera. AB - With the increase in the number of large, 3D, high-resolution nucleic acid structures, particularly of the 30S and 50S ribosomal subunits and the intact bacterial ribosome, advancements in the visualization of nucleic acid structural features are essential. Large molecular structures are complicated and detailed, and one goal of visualization software is to allow the user to simplify the display of some features and accent others. We describe an extension to the UCSF Chimera molecular visualization system for the purpose of displaying and highlighting nucleic acid characteristics, including a new representation of sugar pucker, several options for abstraction of base geometries that emphasize stacking and base pairing, and an adaptation of the ribbon backbone to accommodate the nucleic acid backbone. Molecules are displayed and manipulated interactively, allowing the user to change the representations as desired for small molecules, proteins and nucleic acids. This software is available as part of the UCSF Chimera molecular visualization system and thus is integrated with a suite of existing tools for molecular graphics. PMID- 16478717 TI - Top-down protein sequencing and MS3 on a hybrid linear quadrupole ion trap orbitrap mass spectrometer. AB - Top-down proteomics, the analysis of intact proteins (instead of first digesting them to peptides), has the potential to become a powerful tool for mass spectrometric protein characterization. Requirements for extremely high mass resolution, accuracy, and ability to efficiently fragment large ions have often limited top-down analyses to custom built FT-ICR mass analyzers. Here we explore the hybrid linear ion trap (LTQ)-Orbitrap, a novel, high performance, and compact mass spectrometric analyzer, for top-down proteomics. Protein standards from 10 to 25 kDa were electrosprayed into the instrument using a nanoelectrospray chip. Resolving power of 60,000 was ample for isotope resolution of all protein charge states. We achieved absolute mass accuracies for intact proteins between 0.92 and 2.8 ppm using the "lock mass" mode of operation. Fifty femtomole of cytochrome c applied to the chip resulted in spectra with excellent signal-to-noise ratio and only low attomole sample consumption. Different protein charge states were dissociated in the LTQ, and the sensitivity of the orbitrap allowed routine, high resolution, and high mass accuracy fragment detection. This resulted in unambiguous charge state determination of fragment ions and identification of unmodified and modified proteins by database searching. Protein fragments were further isolated and fragmented in the LTQ followed by analysis of MS(3) fragments in the orbitrap, localizing modifications to part of the sequence and helping to identify the protein with these small peptide-like fragments. Given the ready availability and ease of operation of the LTQ-Orbitrap, it may have significant impact on top-down proteomics. PMID- 16478716 TI - Role of the silkworm argonaute2 homolog gene in double-strand break repair of extrachromosomal DNA. AB - The argonaute protein family provides central components for RNA interference (RNAi) and related phenomena in a wide variety of organisms. Here, we isolated, from a Bombyx mori cell, a cDNA clone named BmAGO2, which is homologous to Drosophila ARGONAUTE2, the gene encoding a repressive factor for the recombination repair of extrachromosomal double-strand breaks (DSBs). RNAi mediated silencing of the BmAGO2 sequence markedly increased homologous recombination (HR) repair of DSBs in episomal DNA, but had no effect on that in chromosomes. Moreover, we found that RNAi for BmAGO2 enhanced the integration of linearized DNA into a silkworm chromosome via HR. These results suggested that BmAgo2 protein plays an indispensable role in the repression of extrachromosomal DSB repair. PMID- 16478718 TI - Interaction of PDZRhoGEF with microtubule-associated protein 1 light chains: link between microtubules, actin cytoskeleton, and neuronal polarity. AB - Rat (r) PDZRhoGEF, initially identified as a glutamate transporter EAAT4 associated protein, is a member of a novel RhoGEF subfamily. The N terminus of the protein contains a PDZ and a proline-rich domain, two motifs known to be involved in protein-protein interactions. By using the yeast two-hybrid approach, we screened for proteins that interact with the N terminus of rPDZRhoGEF. The light chain 2 of microtubule-associated protein 1 (LC2) was the only protein identified from the screen that does not contain a type I PDZ-binding motif at its extreme C terminus (-(S/T)Xphi-COOH, where phi is a hydrophobic amino acid). However, the C terminus does conform to a type II-binding motif (-phiXphi). We report here that rPDZRhoGEF interacts with LC2 via the PDZ domain, and the interaction is abolished by mutations in the carboxylate-binding loop. The specificity of the interaction was confirmed using GST fusion protein pull-down assays and coimmunoprecipitations. Expression of rPDZRhoGEF mutants that are unable to interact with proteins via the carboxylate-binding loop induced changes in cell morphology and actin organization. These mutants alter the activation of RhoGTPases, and coexpression of dominant-negative RhoGTPases prevent the morphological changes. Furthermore, in cells expressing wild type rPDZRhoGEF, drug-induced microtubule depolymerization produces changes in cell morphology that are similar to those induced by rPDZRhoGEF mutants. These results indicate that modulation of the guanine nucleotide exchange activity of rPDZRhoGEF through interaction with microtubule-associated protein light chains may coordinate microtubule integrity and the reorganization of actin cytoskeleton. This coordinated action of the actin and microtubular cytoskeletons is essential for the development and maintenance of neuronal polarity. PMID- 16478719 TI - Vitamin D receptor agonists specifically modulate the volume of the ligand binding pocket. AB - Existing crystal structure data has indicated that 1alpha,25-dihydroxyvitamin D(3) (1alpha,25(OH)(2) D(3)) and its analogues bind the ligand-binding pocket (LBP) of the human vitamin D receptor in a very similar fashion. Because docking of a ligand into the LBP is a more flexible process than crystallography can monitor, we analyzed 1alpha,25(OH)(2)D(3), its 20-epi derivative MC1288, the two side-chain analogues Gemini and Ro43-83582 (a hexafluoro-derivative) by molecular dynamics simulations in a complex with the vitamin D receptor ligand-binding domain and a co-activator peptide. Superimposition of the structures showed that the side chain of MC1288, the first side chain of the conformation II of Gemini, the second side chain of Ro43-83582 in conformation I and the first side chain of Ro43-83582 in conformation II take the same agonistic position as the side chain of 1alpha,25(OH)(2)D(3). Compared with the LBP of the natural hormone MC1288 reduced the volume by 17%, and Gemini expanded it by 19%. The shrinking of the LBP of MC1288 and its expansion to accommodate the second side chain of Gemini or Ro43-83582 is the combined result of minor movements of more than 30 residues and major movements of a few critical amino acids. The agonist-selective recognition of anchoring OH groups by the conformational flexible residues Ala-303, Leu-309, and His-397 was confirmed by in vitro assays. In summary, variations in the volume of agonists lead to adaptations in the volume of the LBP and alternative contacts of anchoring OH-groups. PMID- 16478720 TI - Dysfunctions of cellular oxidative metabolism in patients with mutations in the NDUFS1 and NDUFS4 genes of complex I. AB - The pathogenic mechanism of a G44A nonsense mutation in the NDUFS4 gene and a C1564A mutation in the NDUFS1 gene of respiratory chain complex I was investigated in fibroblasts from human patients. As previously observed the NDUFS4 mutation prevented complete assembly of the complex and caused full suppression of the activity. The mutation (Q522K replacement) in NDUFS1 gene, coding for the 75-kDa Fe-S subunit of the complex, was associated with (a) reduced level of the mature complex, (b) marked, albeit not complete, inhibition of the activity, (c) accumulation of H(2)O(2) and O(2)(.-) in mitochondria, (d) decreased cellular content of glutathione, (e) enhanced expression and activity of glutathione peroxidase, and (f) decrease of the mitochondrial potential and enhanced mitochondrial susceptibility to reactive oxygen species (ROS) damage. No ROS increase was observed in the NDUFS4 mutation. Exposure of the NDUFS1 mutant fibroblasts to dibutyryl-cAMP stimulated the residual NADH-ubiquinone oxidoreductase activity, induced disappearance of ROS, and restored the mitochondrial potential. These are relevant observations for a possible therapeutical strategy in NDUFS1 mutant patients. PMID- 16478721 TI - Dystrophin and utrophin bind actin through distinct modes of contact. AB - This study was designed to define the molecular epitopes of dystrophin-actin interaction and to directly compare the actin binding properties of dystrophin and utrophin. According to our data, dystrophin and utrophin both bound alongside actin filaments with submicromolar affinities. However, the molecular epitopes involved in actin binding differed between the two proteins. In utrophin, the amino-terminal domain and an adjacent string of the first 10 spectrin-like repeats more fully recapitulated the activities measured for full-length protein. The homologous region of dystrophin bound actin with low affinity and near 1:1 stoichiometry as previously measured for the isolated amino-terminal, tandem (CH) domain. In contrast, a dystrophin construct including a cluster of basic spectrin like repeats and spanning from the amino terminus through repeat 17, bound actin with properties most similar to full-length dystrophin. Dystrophin and utrophin both stabilized preformed actin filaments from forced depolymerization with similar efficacies but did not appear to compete for binding sites on actin. We also found that dystrophin binding to F-actin was markedly sensitive to increasing ionic strength, although utrophin binding was unaffected. Although dystrophin and utrophin are functionally homologous actin-binding proteins, these results indicate that their respective modes of contact with actin filaments are markedly different. Finally, we reassessed the abundance of dystrophin in striated muscle using full-length protein as the standard and measured greater than 10-fold higher values than previously reported. PMID- 16478722 TI - Phospholipid transfer activity of microsomal triacylglycerol transfer protein is sufficient for the assembly and secretion of apolipoprotein B lipoproteins. AB - Human microsomal triacylglycerol transfer protein (hMTP) is essential for apolipoprotein B (apoB)-lipoprotein assembly and secretion and is known to transfer triacylglycerols, cholesterol esters, and phospholipids. To understand the relative importance of each lipid transfer activity, we compared the ability of hMTP and its Drosophila ortholog (dMTP) to assemble apoB lipoproteins and to transfer various lipids. apoB48 secretion was induced when co-expressed with either hMTP or dMTP in COS cells, and oleic acid supplementation further augmented secretion without altering particle density. C-terminal epitope-tagged dMTP (dMTP-FLAG) facilitated the secretion of apoB polypeptides in the range of apoB48 to apoB72 but was approximately 50% as efficient as hMTP-FLAG. Comparison of lipid transfer activities revealed that although phospholipid transfer was similar in both orthologs, dMTP was unable to transfer neutral lipids. We conclude that the phospholipid transfer activity of MTP is sufficient for the assembly and secretion of primordial apoB lipoproteins and may represent its earliest function evolved for the mobilization of lipid in invertebrates. Identification of MTP inhibitors, which selectively affect transfer of a specific lipid class, may have therapeutic potential. PMID- 16478723 TI - Functional IsK/KvLQT1 potassium channel in a new corticosteroid-sensitive cell line derived from the inner ear. AB - Endolymph, a high K(+)/low Na(+) fluid, participates in mechanoelectrical transduction in inner ear. Molecular mechanisms controlling endolymph ion homeostasis remain elusive, hampered by the lack of appropriate cellular models. We established an inner ear cell line by targeted oncogenesis. The expression of SV40 T antigen was driven by the proximal promoter of the human mineralocorticoid receptor (MR) gene, a receptor expressed in the inner ear. The EC5v cell line, microdissected from the semicircular canal, grew as a monolayer of immortalized epithelial cells forming domes. EC5v cells exhibited on filters of high transepithelial resistance and promoted K(+) secretion and Na(+) absorption. Functional MR and the 11beta-hydroxysteroid dehydrogenase type 2, a key enzyme responsible for MR selectivity were identified. Expression of the epithelial sodium channel and serum glucocorticoid-regulated kinase 1 was shown to be up regulated by aldosterone, indicating that EC5v represents a novel corticosteroid sensitive cell line. Ionic measurements and (86)Rb transport assays revealed an apical secretion of K(+) at least in part through the I(sK)/KvLQT1 potassium channel under standard culture conditions. However, when cells were exposed to high apically K(+)/low Na(+) fluid, mimicking endolymph exposure, I(sK)/KvLQT1 actually functioned as a strict apical to basolateral K(+) channel inhibited by clofilium. Quantitative reverse transcriptase-PCR further demonstrated that expression of KvLQT1 but not of I(sK) was down-regulated by high K(+) concentration. This first vestibular cellular model thus constitutes a valuable system to further investigate the molecular mechanisms controlling ionic transports in the inner ear and the pathophysiological consequences of their dysfunctions in vertigo and hearing loss. PMID- 16478724 TI - Neutralization of the aspartic acid residue Asp-367, but not Asp-454, inhibits binding of Na+ to the glutamate-free form and cycling of the glutamate transporter EAAC1. AB - Substrate transport by the plasma membrane glutamate transporter EAAC1 is coupled to cotransport of three sodium ions. One of these Na(+) ions binds to the transporter already in the absence of glutamate. Here, we have investigated the possible involvement of two conserved aspartic acid residues in transmembrane segments 7 and 8 of EAAC1, Asp-367 and Asp-454, in Na(+) cotransport. To test the effect of charge neutralization mutations in these positions on Na(+) binding to the glutamate-free transporter, we recorded the Na(+)-induced anion leak current to determine the K(m) of EAAC1 for Na(+). For EAAC1(WT), this K(m) was determined as 120 mm. When the negative charge of Asp-367 was neutralized by mutagenesis to asparagine, Na(+) activated the anion leak current with a K(m) of about 2 m, indicating dramatically impaired Na(+) binding to the mutant transporter. In contrast, the Na(+) affinity of EAAC1(D454N) was virtually unchanged compared with the wild type transporter (K(m) = 90 mm). The reduced occupancy of the Na(+) binding site of EAAC1(D367N) resulted in a dramatic reduction in glutamate affinity (K(m) = 3.6 mm, 140 mm [Na(+)]), which could be partially overcome by increasing extracellular [Na(+)]. In addition to impairing Na(+) binding, the D367N mutation slowed glutamate transport, as shown by pre-steady-state kinetic analysis of transport currents, by strongly decreasing the rate of a reaction step associated with glutamate translocation. Our data are consistent with a model in which Asp-367, but not Asp-454, is involved in coordinating the bound Na(+) in the glutamate-free transporter form. PMID- 16478725 TI - Interrelated roles for Mcl-1 and BIM in regulation of TRAIL-mediated mitochondrial apoptosis. AB - The current study demonstrates a novel cross-talk mechanism between the TRAIL receptor death signaling pathway and the mitochondria. This newly identified pathway is regulated at the mitochondrial outer membrane by a complex between the prosurvival Bcl-2 member, Mcl-1 and the BH3-only protein, Bim. Under non apoptotic conditions, Bim is sequestered by Mcl-1. Direct degradation of Mcl-1 by TRAIL-activated caspase-8 or caspase-3 produces Mcl-1-free Bim that mediates a Bax-dependent apoptotic cascade. Using Mcl-1 or Bim RNAi, we demonstrate that a loss in Mcl-1 expression significantly enhances the mitochondrial apoptotic response to TRAIL that is now mediated by freed Bim. Whereas overexpression of Mcl-1 contributes to the preservation of the mitochondrial membrane potential, Mcl-1 knockdown facilitates the Bim-mediated dissipation of this potential. Loss of Mcl-1 contributes to an increased level of caspase activity downstream of the mitochondrial response to TRAIL. Furthermore, the Mcl-1 expression level at the mitochondrial outer membrane determines the release efficiency for the apoptogenic proteins cytochrome c, Smac, and HtrA2 in response to Bim. These are the first findings to demonstrate the involvement of Bim in the TRAIL-mediated mitochondrial cascade. They also suggest that Mcl-1 may serve as a direct substrate for TRAIL-activated caspases implying the existence of a novel TRAIL/caspase-8/Mcl-1/Bim communication mechanism between the extrinsic and the intrinsic apoptotic pathways. PMID- 16478727 TI - Unique substrate recognition by botulinum neurotoxins serotypes A and E. AB - Botulinum neurotoxins (BoNTs) are zinc proteases that cleave SNARE proteins to elicit flaccid paralysis by inhibiting the fusion of neurotransmitter-carrying vesicles to the plasma membrane of peripheral neurons. There are seven serotypes of BoNT, termed A-G. BoNT serotype A and serotype E cleave SNAP25 at residues 197 198 and 180-181, respectively. Unlike other zinc proteases, the BoNTs recognize extended regions of SNAP25 for cleavage. The basis for this extended substrate recognition and specificity is unclear. Saturation mutagenesis and deletion mapping identified residues 156-202 of SNAP25 as the optimal cleavage domain for BoNT/A, whereas the optimal cleavage domain for BoNT/E was shorter, comprising residues 167-186 of SNAP25. Two sub-sites were resolved within each optimal cleavage domain, which included a recognition or active site (AS) domain that contained the site of cleavage and a binding (B) domain, which contributed to substrate affinity. Within the AS domains, the P1', P3, and P5 sites of SNAP25 contributed to scissile bond cleavage by LC/A, whereas the P1' and P2 sites of SNAP25 contributed to scissile bond cleavage by LC/E. These studies provide insight into the development of strategies for small molecule inhibitors of the BoNTs. PMID- 16478726 TI - TEDS site phosphorylation of the yeast myosins I is required for ligand-induced but not for constitutive endocytosis of the G protein-coupled receptor Ste2p. AB - The yeast myosins I Myo3p and Myo5p have well established functions in the polarization of the actin cytoskeleton and in the endocytic uptake of the G protein-coupled receptor Ste2p. A number of results suggest that phosphorylation of the conserved TEDS serine of the myosin I motor head by the Cdc42p activated p21-activated kinases Ste20p and Cla4p is required for the organization of the actin cytoskeleton. However, the role of this signaling cascade in the endocytic uptake has not been investigated. Interestingly, we find that Myo5p TEDS site phosphorylation is not required for slow, constitutive endocytosis of Ste2p, but it is essential for rapid, ligand-induced internalization of the receptor. Our results strongly suggest that a kinase activates the myosins I to sustain fast endocytic uptake. Surprisingly, however, despite the fact that only p21-activated kinases are known to phosphorylate the conserved TEDS site, we find that these kinases are not essential for ligand-induced internalization of Ste2p. Our observations indicate that a different signaling cascade, involving the yeast homologues of the mammalian PDK1 (3-phosphoinositide-dependent-protein kinase-1), Phk1p and Pkh2p, and serum and glucocorticoid-induced kinase, Ypk1p and Ypk2p, activate Myo3p and Myo5p for their endocytic function. PMID- 16478728 TI - The properties of the positively charged loop region in PSI-G are essential for its "spontaneous" insertion into thylakoids and rapid assembly into the photosystem I complex. AB - The PSI-G subunit of photosystem I (PSI) is an 11-kDa membrane protein that plays an important role in electron transport between plastocyanin and PSI and is involved in the stability of the PSI complex. Within the complex, the PSI-G subunit is bound to PSI-B and is in contact with Lhca1. PSI-G has two transmembrane spans connected by a positively charged stromal loop. The loop is inaccessible to proteases, indicating a tightly bound location within the PSI complex. Here, we have studied the insertion mechanism and assembly of PSI-G. We show that the protein inserts into thylakoids by a direct or "spontaneous" pathway that does not involve the activities of any known chloroplast protein targeting machinery. Surprisingly, the positively charged stromal loop region plays a major role in this process. Mutagenesis or deletions within this region almost invariably lead to a marked lowering of insertion efficiency, strongly indicating a critical role for the loop in the organization of the transmembrane regions prior to or during membrane insertion. Finally, we have examined the assembly of newly inserted PSI-G into the PSI complex, since very little is known of the assembly pathway for this large multimeric complex. Interestingly, we find that inserted PSI-G can be found within the full PSI complex within the import assay time frame after insertion into thylakoids, strongly suggesting that PSI-G normally associates at the end of the assembly process. This is consistent with its location on the periphery of the complex. PMID- 16478729 TI - MarA-mediated transcriptional repression of the rob promoter. AB - The Escherichia coli transcriptional regulator MarA affects functions that include antibiotic resistance, persistence, and survival. MarA functions as an activator or repressor of transcription utilizing similar degenerate DNA sequences (marboxes) with three different binding site configurations with respect to the RNA polymerase-binding sites. We demonstrate that MarA down regulates rob transcripts both in vivo and in vitro via a MarA-binding site within the rob promoter that is positioned between the -10 and -35 hexamers. As for the hdeA and purA promoters, which are repressed by MarA, the rob marbox is also in the "backward" orientation. Protein-DNA interactions show that SoxS and Rob, like MarA, bind the same marbox in the rob promoter. Electrophoretic mobility shift analyses with a MarA-specific antibody demonstrate that MarA and RNA polymerase form a ternary complex with the rob promoter DNA. Transcription experiments in vitro and potassium permanganate footprinting analysis show that MarA affects the RNA polymerase-mediated closed to open complex formation at the rob promoter. PMID- 16478731 TI - The crystal structure of H-2D(b) complexed with a partial peptide epitope suggests a major histocompatibility complex class I assembly intermediate. AB - In the absence of bound peptide ligands, major histocompatibility complex (MHC) class I molecules are unstable. In an attempt to determine the minimum requirement for peptide-dependent MHC class I stabilization, we have used short synthetic peptides derived from the Sendai virus nucleoprotein epitope (residues 324-332, 1FAPGNYPAL9) to promote its folding in vitro of H-2D(b). We found that H 2D(b) can be stabilized by the pentapeptide 5NYPAL9, which is equivalent to the C terminal portion of the optimal nonapeptide and includes both the P5 and P9 anchor residues. We have crystallized the complex of the H-2D(b) molecule with the pentamer and determined the structure to show how a quasi-stable MHC class I molecule can be formed by occupancy of a single binding pocket in the peptide binding groove. PMID- 16478730 TI - Prion proteins with insertion mutations have altered N-terminal conformation and increased ligand binding activity and are more susceptible to oxidative attack. AB - We compared the biochemical properties of a wild type recombinant normal human cellular prion protein, rPrP(c), with a recombinant mutant human prion protein that has three additional octapeptide repeats, rPrP(8OR). Monoclonal antibodies that are specific for the N terminus of rPrP(c) react much better with rPrP(8OR) than rPrP(c), suggesting that the N terminus of rPrP(8OR) is more exposed and hence more available for antibody binding. The N terminus of PrP(c) contains a glycosaminoglycan binding motif. Accordingly, rPrP(8OR) also binds more glycosaminoglycan than rPrP(c). In addition, the divalent cation copper modulates the conformations of rPrP(c) and rPrP(8OR) differently. When compared with rPrP(c), rPrP(8OR) is also more susceptible to oxidative damage. Furthermore, the abnormalities associated with rPrP(8OR) are recapitulated, but even more profoundly, in another insertion mutant, which has five extra octapeptide repeats, rPrP(10OR). Therefore, insertion mutants appear to share common features, and the degree of abnormality is proportional to the number of insertions. Any of these anomalies may contribute to the pathogenesis of inherited human prion disease. PMID- 16478732 TI - The VP2/VP3 minor capsid protein of simian virus 40 promotes the in vitro assembly of the major capsid protein VP1 into particles. AB - The SV40 capsid is composed primarily of 72 pentamers of the VP1 major capsid protein. Although the capsid also contains the minor capsid protein VP2 and its amino-terminally truncated form VP3, their roles in capsid assembly remain unknown. An in vitro assembly system was used to investigate the role of VP2 in the assembly of recombinant VP1 pentamers. Under physiological salt and pH conditions, VP1 alone remained dissociated, and at pH 5.0, it assembled into tubular structures. A stoichiometric amount of VP2 allowed the assembly of VP1 pentamers into spherical particles in a pH range of 7.0 to 4.0. Electron microscopy observation, sucrose gradient sedimentation analysis, and antibody accessibility tests showed that VP2 is incorporated into VP1 particles. The functional domains of VP2 important for VP1 binding and for enhancing VP1 assembly were further explored with a series of VP2 deletion mutants. VP3 also enhanced VP1 assembly, and a region common to VP2 and VP3 (amino acids 119-272) was required to promote VP1 pentamer assembly. These results are relevant for controlling recombinant capsid formation in vitro, which is potentially useful for the in vitro development of SV40 virus vectors. PMID- 16478733 TI - Polo box domain of Plk3 functions as a centrosome localization signal, overexpression of which causes mitotic arrest, cytokinesis defects, and apoptosis. AB - Polo-like kinase 3 (Plk3), an immediate early response gene product, plays an important role in the regulation of mitosis, DNA damage checkpoint activation, and Golgi dynamics. Similar to other members of the Plk family, Plk3 has a conserved kinase domain at the N terminus and a Polo box domain consisting of two Polo boxes at the C terminus. In this study, we demonstrate that the Polo box domain of Plk3 is sufficient for subcellular localization of this kinase to the centrosomes, the spindle poles, and the midbody when ectopically expressed in HeLa and U2OS cells. Both Polo boxes are required for the subcellular localization. Overexpression of the Polo box domain, not the kinase domain, of Plk3 causes significant cell cycle arrest and cytokinesis defects, eventually leading to mitotic catastrophe/apoptosis. Interestingly, the Polo box domain of Plk3 is more potent in inhibiting cell proliferation and inducing apoptosis than that of Plk1, suggesting that this domain can provide an additional structural basis for discovery of new anticancer drugs given the current emphasis on Plk1 as a therapeutic target. PMID- 16478734 TI - The promise of personalized medicine: a conversation with Michael Svinte. Interview by Michael Millenson. AB - In early 2004, IBM combined its Healthcare unit, which focused on the technology needs of providers, with its Life Sciences unit, which catered to research scientists. Out of that union was born an "emerging business opportunity" called information-based medicine, in which IBM has already invested tens of millions in the expectation of reaping billions of dollars in revenues. Michael Svinte describes his mission as providing the information technology infrastructure that will enable technologies such as proteomics and molecular imaging to progress from the bench to the bedside, thereby resulting in predictive and personalized health care. PMID- 16478735 TI - Redox modulation of chemotherapy-induced tumor cell killing and normal tissue toxicity. PMID- 16478736 TI - Vitamin E in the prevention of prostate cancer: where are we today? PMID- 16478738 TI - Changes proposed for reporting chemical spills. PMID- 16478737 TI - Researchers exploring faster alternatives to 2-year test for carcinogenicity. PMID- 16478739 TI - Micronutrients: to supplement, or not to supplement? PMID- 16478740 TI - Criticism of tumor response criteria raises trial design questions. PMID- 16478741 TI - Stat bite: Percent of adults at healthy weight, overweight, or obese, 1971-2002. PMID- 16478742 TI - Improvement of the therapeutic index of anticancer drugs by the superoxide dismutase mimic mangafodipir. AB - BACKGROUND: Anticancer drugs act by increasing intracellular hydrogen peroxide levels. Mangafodipir, a superoxide dismutase (SOD) mimic with catalase and glutathione reductase activities, protects normal cells from apoptosis induced by H2O2. We investigated its and other oxidative stress modulators' effects on anticancer drug activity in vitro and in vivo. METHODS: Cell lysis and intracellular reactive oxygen species levels were assessed in vitro in human leukocytes from healthy subjects and in murine CT26 colon cancer cells. Cells were exposed to the chemotherapeutic agents paclitaxel, oxaliplatin, or 5 fluorouracil, either in the presence or absence of mangafodipir and other oxidative stress modulators. Cell viability was evaluated by the methylthiazoletetrazolium assay. The effects of mangafodipir and other oxidative stress modulators on peripheral blood counts and on tumor growth were studied in BALB/c mice that were implanted with CT26 tumors and treated with 20 mg/kg paclitaxel. Survival of BALB/c mice infected with Staphylococcus aureus was also examined by treatment group. Statistical tests were two-sided. RESULTS: In vitro lysis of leukocytes exposed to paclitaxel, oxaliplatin, or 5-fluorouracil in combination with mangafodipir was decreased by 46% (95% confidence interval [CI] = 44% to 48%), 30.5% (95% CI = 29% to 32%), and 15% (95% CI = 10% to 20%), compared with lysis of cells treated with anticancer agent alone. Mangafodipir also statistically significantly enhanced in vitro anticancer drug cytotoxicity toward CT26 cancer cells. In vivo, mangafodipir protected mice against paclitaxel induced leukopenia. Moreover, the survival rate of mice infected with S. aureus and treated with paclitaxel was higher when mangafodipir was also administered (survival: 3 of 17 versus 14 of 17, P < .001). In addition, mangafodipir amplified the inhibitory effect of paclitaxel on CT26 tumor growth in mice. CONCLUSIONS: Mangafodipir decreased hematotoxicity and enhanced cytotoxicity of anticancer agents. PMID- 16478743 TI - Supplemental and dietary vitamin E, beta-carotene, and vitamin C intakes and prostate cancer risk. AB - BACKGROUND: Vitamin E, beta-carotene, and vitamin C are micronutrient antioxidants that protect cells from oxidative damage involved in prostate carcinogenesis. In separate trials, supplemental vitamin E was associated with a decreased risk of prostate cancer among smokers and supplemental beta-carotene was associated with a decreased risk of prostate cancer among men with low baseline plasma beta-carotene levels. METHODS: We evaluated the association between intake of these micronutrient antioxidants from foods and supplements and the risk of prostate cancer among men in the screening arm of the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial. At baseline, trial participants completed a 137-item food frequency questionnaire that included detailed questions on 12 individual supplements. Cox proportional hazards models were used to estimate relative risks (RRs) and 95% confidence intervals (CIs). All statistical tests were two-sided. RESULTS: We identified 1338 cases of prostate cancer among 29 361 men during up to 8 years of follow-up. Overall, there was no association between prostate cancer risk and dietary or supplemental intake of vitamin E, beta-carotene, or vitamin C. However, among current and recent (i.e., within the previous 10 years) smokers, decreasing risks of advanced prostate cancer (i.e., Gleason score > or = 7 or stage III or IV) were associated with increasing dose (RR for > 400 IU/day versus none = 0.29, 95% CI = 0.12 to 0.68; Ptrend = .01) and duration (RR for > or = 10 years of use versus none = 0.30, 95% CI = 0.09 to 0.96; Ptrend = .01) of supplemental vitamin E use. Supplemental beta carotene intake at a dose level of at least 2000 microg/day was associated with decreased prostate cancer risk in men with low (below the median of 4129 microg/day) dietary beta-carotene intake (RR = 0.52, 95% CI = 0.33 to 0.81). Among smokers, the age-adjusted rate of advanced prostate cancer was 492 per 100,000 person-years in those who did not take supplemental vitamin E, 153 per 100,000 person-years in those who took more than 400 IU/day of supplemental vitamin E, and 157 per 100,000 person-years in those who took supplemental vitamin E for 10 or more years. Among men with low dietary beta-carotene intake, the age-adjusted rate of prostate cancer was 1122 per 100,000 person-years in those who did not take supplemental beta-carotene, and 623 per 100,000 person years in those who took at least 2000 microg/day of supplemental beta-carotene. CONCLUSIONS: Our results do not provide strong support for population-wide implementation of high-dose antioxidant supplementation for the prevention of prostate cancer. However, vitamin E supplementation in male smokers and beta carotene supplementation in men with low dietary beta-carotene intakes were associated with reduced risk of this disease. PMID- 16478744 TI - Gene promoter methylation in prostate tumor-associated stromal cells. AB - BACKGROUND: Gene expression can be silenced through the methylation of specific sites in the promoter region. This mechanism of gene silencing has an important role in the carcinogenesis of prostate and other cancers. Although tumor associated stromal cells also exhibit changes in gene expression, promoter methylation has not been described in these cells. METHODS: Tumor epithelia, tumor-associated stroma and normal epithelia, and stroma adjacent to tumor tissues were isolated from whole-mount prostatectomy specimens (two per patient) of patients (n = 5) with localized prostate cancer and from normal epithelia and stroma from benign prostate hyperplasia specimens (two per patient) from men (n = 5) without prostate cancer by using laser capture microdissection or expression microdissection. The methylation status of three genes important in prostate carcinogenesis, GSTP1, RARbeta2, and CD44, were evaluated using quantitative methylation-sensitive polymerase chain reaction. RESULTS: GSTP1 and RARbeta2 were methylated in the tumor epithelium of all five prostate cancer patients and in the tumor-associated stroma in four of the five patients. CD44 was methylated in the tumor epithelium from four of the five patients but not in the tumor stroma. GSTP1 and RARbeta2 were methylated in normal epithelium of two and four patients, respectively, and in normal stroma of one and two patients, respectively, that were isolated from regions adjacent to the tumors and may have resulted from a tumor-field effect; CD44 methylation was not observed in normal epithelium or stroma. In contrast, normal epithelia and stroma from benign prostate hyperplasia specimens showed no promoter methylation in GSTP1, RARbeta2, or CD44. CONCLUSIONS: The observation of promoter methylation in the non-neoplastic cells of the prostate tumor microenvironment may advance our understanding of prostate cancer development and progression and lead to new diagnostic and prognostic markers and therapeutic targets. PMID- 16478745 TI - Gene expression profiling in breast cancer: understanding the molecular basis of histologic grade to improve prognosis. AB - BACKGROUND: Histologic grade in breast cancer provides clinically important prognostic information. However, 30%-60% of tumors are classified as histologic grade 2. This grade is associated with an intermediate risk of recurrence and is thus not informative for clinical decision making. We examined whether histologic grade was associated with gene expression profiles of breast cancers and whether such profiles could be used to improve histologic grading. METHODS: We analyzed microarray data from 189 invasive breast carcinomas and from three published gene expression datasets from breast carcinomas. We identified differentially expressed genes in a training set of 64 estrogen receptor (ER)-positive tumor samples by comparing expression profiles between histologic grade 3 tumors and histologic grade 1 tumors and used the expression of these genes to define the gene expression grade index. Data from 597 independent tumors were used to evaluate the association between relapse-free survival and the gene expression grade index in a Kaplan-Meier analysis. All statistical tests were two-sided. RESULTS: We identified 97 genes in our training set that were associated with histologic grade; most of these genes were involved in cell cycle regulation and proliferation. In validation datasets, the gene expression grade index was strongly associated with histologic grade 1 and 3 status; however, among histologic grade 2 tumors, the index spanned the values for histologic grade 1-3 tumors. Among patients with histologic grade 2 tumors, a high gene expression grade index was associated with a higher risk of recurrence than a low gene expression grade index (hazard ratio = 3.61, 95% confidence interval = 2.25 to 5.78; P < .001, log-rank test). CONCLUSIONS: Gene expression grade index appeared to reclassify patients with histologic grade 2 tumors into two groups with high versus low risks of recurrence. This approach may improve the accuracy of tumor grading and thus its prognostic value. PMID- 16478746 TI - Randomized, double-blind, active-controlled trial of every-3-week darbepoetin alfa for the treatment of chemotherapy-induced anemia. AB - BACKGROUND: In the United States, darbepoetin alfa (Aranesp) is often used to treat patients with chemotherapy-induced anemia using weekly or every-2-week administration schedules. In Europe, darbepoetin alfa is used either weekly or in every-3-week dosing. The every-3-week schedule can be synchronized with many chemotherapy regimens, resulting in fewer visits and reducing burden to patients, but the safety and efficacy of this regimen have not been clear. METHODS: A randomized, double-blind, double-dummy, active-controlled phase 3 trial was performed in 110 European centers. Eligible patients (age > or = 18 years) were anemic (hemoglobin level < 11 g/dL), had a nonmyeloid malignancy, and were to receive at least 12 weeks of chemotherapy. Patients were randomly assigned 1:1 to darbepoetin alfa treatment every 3 weeks (500-microg dose) or weekly (2.25 microg/kg) for 15 weeks. We compared red blood cell transfusion incidence among the two arms from week 5 to the end of the treatment phase using a noninferiority study design. Noninferiority was determined if the upper limit of the 95% confidence interval (CI) for the difference in blood transfusions between groups, calculated using Kaplan-Meier methods, did not exceed 12.5%, a margin based on previous placebo-controlled studies. RESULTS: A total of 705 patients were randomly assigned, and 672 remained in the study at week 5. Fewer patients in the every-3-week arm than in the weekly arm received blood transfusions from week 5 to the end of the treatment phase (unadjusted Kaplan-Meier estimates = 23% versus 30%, difference = -6.8%; 95% CI = -13.6 to 0.1). Percentages of patients achieving the target hemoglobin level (> or = 11 g/dL, consistent with evidence based practice guidelines) were 84% (every 3 weeks) and 77% (weekly). The frequency of cardiovascular/thromboembolic adverse events was 8% in both groups, and safety was comparable. CONCLUSIONS: Patients with chemotherapy-induced anemia can safely and effectively be treated with 500 microg of darbepoetin alfa every 3 weeks. PMID- 16478747 TI - Association between MDM2-SNP309 and age at colorectal cancer diagnosis according to p53 mutation status. AB - A single-nucleotide polymorphism (SNP) in the promoter of the MDM2 gene, SNP309 (a T-->G change), was recently implicated in the early onset of cancer in individuals with Li-Fraumeni syndrome and of sporadic soft-tissue sarcoma. SNP309 induces an increase in the level of Mdm2 protein, which causes attenuation of the p53 pathway. To investigate the effect of this polymorphism in colorectal cancer pathogenesis, we genotyped 153 colorectal cancer patients who were randomly selected from among 330 consecutive patients stratified according to p53 mutation status and age at diagnosis, for alleles of MDM2-SNP309. Among the 77 patients with p53 wild-type tumors, the median age at colorectal cancer diagnosis was 71.5 years for patients with the T/T genotype and 61.0 years for patients with SNP309 (T/G or G/G genotype) (estimated difference between medians [Hodges-Lehmann method] = 8.0 years, 95% confidence interval = 1.0 to 16.0 years; P = .03 [two sided Wilcoxon rank sum test]). Our data indicate that MDM2-SNP309 is a modifier of the age at colorectal cancer onset for patients whose tumors have a wild-type p53 gene. PMID- 16478748 TI - Beta-blockers and heart failure in older people. PMID- 16478749 TI - Incidence of recognized and unrecognized myocardial infarction in men and women aged 55 and older: the Rotterdam Study. AB - AIMS: Contemporary data on the incidence of unrecognized myocardial infarction (MI) among subjects aged 55 and older are limited. METHODS AND RESULTS: We studied the incidence of recognized and unrecognized MI in the Rotterdam Study, a population-based cohort of men and women aged 55 and older. The baseline examination was performed during 1990-93, with follow-up examinations during 1994 95, and 1997-2000. Baseline and follow-up 12-lead ECGs were analysed by the Modular ECG Analysis System. The 5148 participants who had no evidence of prevalent infarction were the subjects for analysis. Incident recognized infarction was defined as the occurrence of a fatal or non-fatal event coded as I21 according to the International Classification of Diseases, 10th edition. A repeat ECG was available in 4187 subjects. An unrecognized infarction was considered to have occurred if there was electrocardiographic evidence in the absence of a clinically recognized event. During a median follow-up of 6.4 years, 141 incident recognized infarctions occurred and the incidence rate of this event was 5.0 per 1000 person years. The incidence was higher in men (8.4) than in women (3.1). The incidence rate of unrecognized infarction was 3.8 per 1000 person years. Men (4.2) and women (3.6) had approximately similar incidence. Hence, the proportion of unrecognized infarction was lower in men (33%) than in women (54%). This difference in proportion of unrecognized infarctions was independent of age. CONCLUSION: A high proportion of incident MIs remains clinically unrecognized. As a history of MI is associated with an increased risk of repeat cardiovascular complications, our data suggest a need for periodical electrocardiographic screening to recognize (prevalent) infarctions and to install effective preventive treatment in those aged 55 and older. PMID- 16478750 TI - Role of the SPIRAL1 gene family in anisotropic growth of Arabidopsis thaliana. AB - Arabidopsis spiral1 (spr1) mutants show a right-handed helical growth phenotype in roots and etiolated hypocotyls due to impaired directional growth of rapidly expanding cells. SPR1 encodes a small protein with as yet unknown biochemical functions, though its localization to cortical microtubules (MTs) suggests that SPR1 maintains directional cell expansion by regulating cortical MT functions. The Arabidopsis genome contains five SPR1-LIKE (SP1L) genes that share high sequence identity in N- and C-terminal regions. Overexpression of SP1Ls rescued the helical growth phenotype of spr1, indicating that SPR1 and SP1L proteins share the same biochemical functions. Expression analyses revealed that SPR1 and SP1L genes are transcribed in partially overlapping tissues. A combination of spr1 and sp1l mutations resulted in randomly oriented cortical MT arrays and isotropic expansion of epidermal cells. These observations suggest that SPR1 and SP1Ls act redundantly in maintaining the cortical MT organization essential for anisotropic cell growth, and that the helical growth phenotype of spr1 results from a partially compromised state of cortical MTs. Additionally, inflorescence stems of spr1 sp1l multiple mutants showed a right-handed tendril-like twining growth, indicating that a directional winding response may be conferred to the non-directional nutational movement by modulating the expression of SPR1 homologs. PMID- 16478751 TI - Enhanced stability of recombinant keratinocyte growth factor by mutagenesis. AB - Native sequence keratinocyte growth factor (KGF) is fairly unstable, as manifested by the loss of the monomeric native protein accompanied by the accumulation of aggregated species during storage at moderate temperatures. Several different types of analogs were generated and the storage stability of the protein assessed. In the first type of analog one or more of the five cysteinyl residues in KGF were replaced; in the second class the N-terminal residues that included the first disulfide bond were deleted. Both of these types of analogs involved removal of the disulfide bond between cysteines 1 and 15. The third group involved mutating one of the basic amino acids located in a cluster of positive charges (involved in heparin binding) around Arg144 to a neutral or acidic amino acyl residue. Among the cysteine replacement analogs, the double mutation of Cys1 and 15 to Ser resulted in significantly increased stability without compromising the mitogenic activity, while Cys to Ser mutations at other positions were either destabilizing or had no effect. Deletion of the 15, 23 or 27 N-terminal amino acyl residues also increased the stability of the protein. The activity of the analogs was not affected by the deletion of 15 or 23 amino acids, but it was significantly decreased upon removal of the 27 N-terminal amino acyl residues. Much greater stability was achieved by mutation of the basic amino acids, especially Arg144, to Glu or Gln, but this increase in stability was accompanied by large decrease in activity. The analog with the 23 N-terminal amino acyl residues deleted represents one of the best compromises between increased stability and retention of activity. PMID- 16478752 TI - The impact of experimental design on assessing mechanism-based inactivation of CYP2D6 by MDMA (Ecstasy). AB - MDMA (3-4-methylenedioxymethamphetamine, commonly known as Ecstasy) is a potent mechanism-based inhibitor (MBI) of cytochrome P450 2D6 (CYP2D6), causing quasi irreversible inhibition of the enzyme in vitro. An evaluation of the in vivo implications of this phenomenon depends on the accuracy of the estimates of the parameters that define the inhibition in vitro, namely k(inact) (the maximal inhibition rate) and KI (the inactivation constant). These values are determined in two steps, pre-incubation of the enzyme with the inhibitor (enzyme inactivation), followed by dilution and further incubation to measure residual enzyme activity with a probe substrate. The aim of this study was to assess the impact of different dilutions and probe substrate concentrations on the estimates of k(inact) and KI using recombinantly expressed CYP2D6. Enzyme activity was measured by the conversion of dextromethorphan (DEX) to dextrorphan (DOR). Dilution factors of 1.25, 2, 5, 10, 25 and 50 (DEX at 30 microM) gave mean (+/ SE) values of k(inact) (min-1) of 0.20+/-0.06, 0.21+/-0.05, 0.31+/-0.06, 0.37+/ 0.11, 0.51+/-0.10 and 0.58+/-0.08, respectively, and KI (microM) values (after correction for non-specific microsomal binding) of 2.22+/-1.90, 2.80+/-1.34, 5.78+/-2.07, 6.36+/-2.93, 3.99+/-1.57 and 4.86+/-1.37, respectively. Accordingly, high (e.g. 50 fold) and low (e.g. 1.25 fold) dilutions were associated with statistically significant differences in kinetic values (p <0.05). Varying DEX concentration (10-100 microM) was not associated with significant changes in k(inact) and KI values when a five-fold dilution was used (with the exception of a lower KI at 10 microM DEX). High dilution was also shown to reduce non-specific microsomal binding of MDMA. The changes in the two kinetic parameters were dependent on the experimental procedure and shown to be unlikely to have a material influence on the maximum inhibition of CYP2D6 expected in vivo after typical recreational doses of MDMA (50-100 mg), since the potency of inhibition was high. The different values of the kinetic parameters were predicted to have a marginal influence on the time for recovery of enzyme activity following re synthesis of CYP2D6. PMID- 16478753 TI - The influence of the CYP2D6 polymorphism on psychopathological and extrapyramidal symptoms in the patients on long-term antipsychotic treatment. AB - Poor response to antipsychotics treatment and extrapyramidal side effects (EPS) are the most challenging problems in the treatment of schizophrenia. Several studies were investigating the impact of polymorphic cytochrome P450 2D6 gene (CYP2D6) on EPS but the results were conflicting. There are practically no clinical studies of long-term treatment of schizophrenia and CYP2D6 polymorphism. Our aim was to evaluate the influence of CYP2D6 genotype on psychopathological symptoms and the occurrence of EPS in Slovenian outpatients with schizophrenia or schizoaffective disorder in stable remission, receiving long-term maintenance antipsychotic treatment. In total 131 outpatients meeting the DSM IV criteria for schizophrenia or schizoaffective disorder and receiving maintenance therapy with haloperidol, fluphenazine, zuclopethixole or risperidone were genotyped for 14 polymorphic CYP2D6 alleles. Psychopathological symptoms were assessed with the Positive and Negative Symptom Scale for Schizophrenia (PANSS). EPS were assessed with the Simpson Angus Scale (SAS), the Barnes Akathisia Scale and the Abnormal Involuntary Movement Scale (AIMS). Six patients (4.6%) were genotyped as poor metabolizers (PMs). PMs scored significantly higher on the negative subscale for PANSS. There were no statistically significant differences between the group of PMs and the group of patients with at least one functional CYP2D6 allele in view of patient's characteristics or any of the items of the AIMS, the SAS or the Barnes Akathisia Scale. CYP2D6 genotype may not be the major factor that determines the susceptibility to antipsychotic-induced EPS in Slovenian patients in stable remission and on maintenance therapy with antipsychotics that are mainly CYP2D6 substrates. However, CYP2D6 genotype might be a factor contributing to the persistent negative symptoms of schizophrenia. PMID- 16478755 TI - No effects of l-dopa and bromocriptine on psychophysiological parameters of human selective attention. AB - Patients with schizophrenia exhibit diverse cognitive deficits, one of which is a loss of the ability to focus attention. According to the revised dopamine hypothesis of schizophrenia both an increased mesolimbic and a decreased prefrontal dopaminergic activity is suggested to be involved in schizophrenia. The current study was designed to explore the relationship between dopamine and two psychophysiological parameters of selective attention, i.e. P300 amplitude and processing negativity (PN) in healthy volunteers. In two separate experiments, with a double-blind, balanced and placebo-controlled crossover design, 18 healthy male volunteers were orally administered either 300 mg l-dopa (precursor of dopamine) or placebo (experiment I), or 1.25mg bromocriptine (D2 agonist) or placebo (experiment II). Following this treatment they were tested in an auditory, dichotic selective attention paradigm. An increase in P300 amplitude was found following deviant stimuli when compared to standard stimuli and following attended stimuli when compared to unattended stimuli, regardless of treatment. Similarly, PN was found regardless of treatment. Neither l-dopa nor bromocriptine affected task performance or the amplitudes of PN or P300. In the present study neither l-dopa nor bromocriptine affected PN, P300 amplitude or task performance in healthy controls, phenomena which are usually found to be disrupted in schizophrenia. This indicates that P300 amplitude and PN are neither affected by a global (l-dopa) increased dopaminergic activity, nor by a more selectively towards striatal areas targeted (bromocriptine) increase in dopaminergic activity. PMID- 16478754 TI - Selective increases in the cytokine, TNFalpha, in the prefrontal cortex of PCP treated rats and human schizophrenic subjects: influence of antipsychotic drugs. AB - The psychotomimetic drug phencyclidine (PCP) induces symptoms closely related to those of schizophrenia in humans. In order to test the hypothesis that cytokines may be involved in the aetiology and treatment of schizophrenia, this study investigated the levels of cytokine mRNAs in rat brain after acute and chronic administration of PCP, in the presence and absence of antipsychotic drugs. The levels of the mRNAs encoding TNF, IL-2, IL-6, TGF 1, 2, 3, IL-3 and GM-CSF were measured in the prefrontal cortex, cortex, hippocampus, ventral and dorsal striatum regions of male hooded Long Evans rats after acute drug administration. Antipsychotic drugs and PCP significantly reduced the levels of TNF in the prefrontal cortex compared to vehicle-treated animals, whilst other cytokines remained unchanged. In addition, significant reductions in the levels of TNF mRNA in the prefrontal cortex still occurred 24h after acute PCP administration. However, levels of TNF mRNA were restored to control values after chronic PCP treatment, whereas increased expression was detected in animals co-administered with haloperidol. Levels of TNF mRNA were also found to be significantly increased in the prefrontal cortex of schizophrenic subjects. The relationship between TNF levels and schizophrenia are discussed. PMID- 16478756 TI - The adult psychiatrist's dilemma: psychostimulant use in attention deficit/hyperactivity disorder. AB - Children who have taken psychostimulants long term for attention deficit/hyperactivity disorder (ADHD) are now presenting at adult psychiatry clinics at an age at which bipolar disorder and schizophrenia may first appear and at which pregnancy is a possibility. Doctors who may have little experience in the management of ADHD or use of psychostimulants are faced with decisions on whether to continue or withdraw medication. A literature search conducted to clarify these issues revealed that, although psychostimulants are generally efficacious in ADHD, the risks of withdrawal reactions and of possible long-term effects such as drug addiction, psychosis and depression, or effects in pregnancy, have not been adequately investigated. In particular, there is little information on the effects of withdrawal and no firm guidelines on methods of psychostimulant discontinuation. There is a need for further research to clarify the pharmacological issues involved in the drug treatment of ADHD and for overlapping arrangements between child and adult psychiatry clinics in the management of ADHD. PMID- 16478757 TI - Cadmium-induced apoptosis in rat kidney epithelial cells involves decrease in nuclear factor-kappa B activity. AB - Renal epithelial cells undergo apoptosis upon exposure to cadmium (Cd). Transcription factors, such as nuclear factor-kappa B (NF-kappaB), mediate the expression of a number of genes involved in apoptosis. The present study was designed to examine the involvement of this transcription factor in Cd-induced apoptosis. Rat kidney proximal tubular epithelial cells, NRK-52E, were incubated with up to 20 microM CdCl2 in serum-free medium for 5 h followed by incubation in serum-containing medium (without Cd) for an additional 12 h. The cells accumulated 582 +/- 19 ng Cd/mg protein after 5-h exposure to 20 microM Cd. As a result of Cd exposure, the DNA-binding activity of the p65 subunit of NF-kappaB was decreased in a concentration- and time-dependent manner. The activity of tumor necrosis factor-alpha-induced inhibitor of kappa B (IkappaB) kinase alpha was also inhibited by Cd. In addition, the phosphorylation of IkappaB-alpha and NF-kappaB p65, as well as the levels of NF-kappaB target gene products, cIAP-1 and cIAP-2, were reduced. Pretreatment of the cells with the antioxidant U83836E or butylated hydroxytoluene preserved the DNA-binding activity and blocked the Cd induced decease in IkappaB-alpha phosphorylation. Cd exposure caused the activation of caspase-3, -7, and -9 and DNA fragmentation. By flow cytometry, 14.6 and 30.5% apoptosis was detected at 6 and 12 h after stopping the Cd exposure. Overexpression of NF-kappaB p65 by transient transfection protected the cells from the Cd-induced apoptosis. Conversely, attenuation of NF-kappaB activity by pretreatment with SN50, an NF-kappaB nuclear translocation inhibitor, potentiated apoptosis. These results suggest that Cd-induced apoptosis involves suppression of NF-kappaB activity which may be mediated by oxidative stress. PMID- 16478758 TI - Attributable risk function in the proportional hazards model for censored time-to event. AB - Time-to-event endpoints are often used in clinical and epidemiological studies to evaluate disease association with hazardous exposures. In the statistical literature of time-to-event analysis, such association is usually measured by the hazard ratio in the proportional hazards model. In public health, it is also of important interest to assess the excess risk attributable to an exposure in a given population. In this article, we extend the notion of 'population attributable fraction' for the binary outcomes to the attributable risk function for the event times in prospective studies. A simple estimator of the time varying attributable risk function is proposed under the proportional hazards model. Its inference procedures are established. Monte-Carlo simulation studies are conducted to evaluate its validity and performance. The proposed methodology is motivated and demonstrated by the data collected in a multicenter acquired immunodeficiency syndrome (AIDS) cohort study to estimate the attributable risk of human immunodeficiency virus type 1 (HIV-1) infections due to several potential risk factors. PMID- 16478759 TI - Cytotrophoblast stem cell lines derived from human embryonic stem cells and their capacity to mimic invasive implantation events. AB - BACKGROUND: An effective embryonic-maternal interaction is crucial for successful human pregnancy. Failure of this process is a major cause of infertility and can lead to placental dysfunction resulting in recurrent miscarriage, fetal retardation and pre-eclampsia. Research is severely constrained by ethical and practical considerations; therefore, we aimed to generate cytotrophoblast stem (CTBS) cell lines from human embryonic stem cells (HESCs). METHOD: Beta-HCG was used as a marker of viable trophoblast cells. In defined culture, embryoid bodies were generated from HESCs and selected for trophoblast enrichment by rounds of cellular aggregation and disaggregation. Distinct CTBS cell lines were isolated and characterized. Spheroid cytotrophoblast bodies were generated and their interaction with luteal-phase endometrial stroma was analysed by real-time image analysis. RESULTS: Three CTBS cell lines were derived, which were maintained in the absence of residual HESCs, fibroblast feeder cells or extracellular matrix. CTBS cells displayed typical cytotrophoblast and syncytiotrophoblast characteristics and exhibited further differentiation to invasive endovascular cell phenotype. One cell line was generated with constitutive expression of enhanced green fluorescent protein (eGFP). Spheroid trophoblast bodies mimicked closely the early invasive stages of implantation when incubated with human endometrial stromal preparations in vitro. CONCLUSION: These human CTBS cell lines are a significant new model for investigating human placentation and may have considerable potential in cell therapy applications. PMID- 16478760 TI - Estrogen receptor 1 haplotype does not regulate oral contraceptive-induced changes in haemostasis and inflammation risk factors for venous and arterial thrombosis. AB - BACKGROUND: Administration of oral contraceptives (OCs) has profound effects on the plasma levels of haemostasis and inflammation variables, resulting in an increased thrombosis risk. Individuals show large differences in the response of these variables to OCs. Polymorphism in the estrogen receptor-1 (ER1) gene may explain part of this inter-individual response. METHODS: We investigated the relationship between variants (c.454-397T>C and c.454-351A>G polymorphisms and the combined haplotype) in the ER1 gene in relation to changes in haemostasis and inflammation variables that are known risk factors for thrombosis in 507 healthy, nonsmoking, nulliparous women receiving six cycles of monophasic OCs with 20, 30 or 50 microg/day estrogen. RESULTS: A significant relationship was observed between the ER1 haplotype and changes in tissue-type plasminogen activator activity (P = 0.006), but no clear interaction pattern between the genotypes or between the estrogen doses was seen. No relationships were observed for the other variables, neither in the haplotype nor in the single polymorphism analysis. CONCLUSION: The ER1 haplotype does not have a strong effect on the estrogen induced changes in haemostasis and inflammation risk markers for arterial and venous thrombosis. PMID- 16478761 TI - Chlamydia trachomatis and chlamydial heat shock protein 60-specific antibody and cell-mediated responses predict tubal factor infertility. AB - BACKGROUND: To evaluate the role of Chlamydia trachomatis-induced humoral and cell-mediated immune (CMI) responses in predicting tubal factor infertility (TFI). METHODS: Blood samples were taken from 88 women with TFI and 163 control women. C. trachomatis and chlamydial heat shock protein 60 (CHSP60)-specific immunoglobulin G (IgG) antibodies were analysed using enzyme-linked immunosorbent assay (ELISA) kits. Proliferative reactivity of peripheral blood mononuclear cells was studied in vitro against Chlamydia elementary body (EB) and recombinant CHSP60 antigens. RESULTS: C. trachomatis-specific IgG antibodies were found more frequently (43.2 versus 13.5%), and the antibody levels were higher in the TFI cases than in the controls (P < 0.001). C. trachomatis EB-induced lymphocyte responses were positive in 81.8% of the TFI cases and 58.9% of the controls (P < 0.001). Similarly, CHSP60-induced lymphocyte responses were found in 45.5% of the TFI cases and 30.7% of the controls (P < 0.001). CHSP60 antibody test was the best single test predicting TFI. Compared to cases with all four markers negative, the estimated risk for TFI was 4.1 (95% CI 1.4-11.9) among those with one positive marker and 19.9 (95% CI 6.9-57.4) among those with three to four positive markers. CONCLUSION: Our results show that TFI prediction model can be improved by combining tests for humoral and CMI response to chlamydial antigens. PMID- 16478762 TI - Effect of local lignocaine gel application for pain relief during suction termination of first-trimester pregnancy: a randomized controlled trial. AB - BACKGROUND: This study evaluated the effectiveness of local lignocaine gel application in pain control during first-trimester suction termination of pregnancy (STOP). METHODS: In this prospective randomized placebo-controlled double-blind trial, 131 women undergoing STOP between 7 and 10 weeks of gestation were studied. They were computer-randomized to receive 2% lignocaine gel or placebo (KY Jelly) locally applied to the cervix 1 min before cervical manipulation/dilatation. They all had cervical priming with misoprostol and premedication with diazepam and pethidine. Pain scores on a verbal analogue scale preoperative, at cervical manipulation/dilatation, intraoperative and 1 h post operative, as well as the patients' satisfaction level towards pain control, were compared. RESULTS: The lignocaine gel group had significantly reduced overall intraoperative pain score compared with placebo group (P = 0.021). No significant difference in pain score between the two groups was demonstrated at other time points. Subgroup analysis revealed that the difference in overall intraoperative pain scores between the two groups was evident in the multiparous (P = 0.015) but not the nulliparous subjects. CONCLUSION: The use of local lignocaine gel application reduces overall intraoperative pain in multiparous women undergoing first-trimester STOP preceded by misoprostol cervical priming and premedication for conscious sedation. PMID- 16478763 TI - Baseline dimensions of the human vagina. AB - BACKGROUND: Vaginal anatomy has been poorly studied. This study aimed to measure baseline dimensions of the undistended vagina of women of reproductive age. METHODS: We combined baseline information collected from five clinical trials using magnetic resonance imaging (MRI) to quantify distribution of a vaginal gel. Seventy-seven MRI scans were performed on 28 women before gel application to establish baseline vaginal measurements. Average dimensions were calculated for each woman and for the population. The influence of potential covariates (age, height, weight and parity) on these dimensions was assessed. RESULTS: MRI measurements are reproducible. The SD surrounding the mean at each anatomical site, and with summary measurements, was significantly smaller with each subject compared with the population. Mean vaginal length from cervix to introitus was 62.7 mm. Vaginal width was largest in the proximal vagina (32.5 mm), decreased as it passed through the pelvic diaphragm (27.8 mm) and smallest at the introitus (26.2 mm). Significant positive associations were parity with vaginal fornix length, age with pelvic flexure width and the height with width at the pelvic flexure. CONCLUSION: No one description characterized the shape of the human vagina. Although there is variation among women, variables such as parity, age and height are positively associated with differences in baseline dimensions. PMID- 16478764 TI - Efficacy of combined metformin-letrozole in comparison with metformin-clomiphene citrate in clomiphene-resistant infertile women with polycystic ovarian disease. AB - BACKGROUND: Adding metformin to clomiphene citrate in clomiphene-resistant polycystic ovary syndrome (PCOS) patients increases ovulatory response. However, because of anti-estrogenic effects of clomiphene it may be associated with lower pregnancy rate, offsetting the ovulation rate benefit. Letrozole is an aromatase inhibitor which induces ovulation without anti-estrogenic effects. METHODS: Infertile women with PCOS were randomly divided into metformin-letrozole (29 patients) and metformin-clomiphene groups (30 patients). After an initial 6-8 weeks of metformin, they received either letrozole (2.5 mg) or clomiphene (100 mg) from day 3-7 of their menstrual cycle. Estradiol (E2) levels, number of follicles, pregnancy rates and endometrial thickness were measured on the day of HCG administration. RESULTS: Mean total E2 and E2 per mature follicle were significantly higher in clomiphene group without a difference in mean number of mature follicles >18 mm and ovulation rate. Endometrial thickness was significantly higher in letrozole group. The pregnancy rate in letrozole group (10 patients, 34.50%) as compared with clomiphene group (5 patients, 16.67%) did not show significant difference, whereas full-term pregnancies were higher in letrozole group [10 patients (34.50%) versus 3 patients (10%)]. CONCLUSION: In clomiphene-resistant PCOS patients, the combination of letrozole and metformin leads to higher full-term pregnancies. PMID- 16478765 TI - Stomatal oscillations in orange trees under natural climatic conditions. AB - BACKGROUND AND AIMS: Stomatal oscillations have been reported in many plant species, but they are usually induced by sudden step changes in the environment when plants are grown under constant conditions. This study shows that in navel orange trees (Citrus sinensis) pronounced stomatal oscillations occur and persist under natural climatic conditions. METHODS: Oscillations in stomatal conductance were measured, and related to simultaneous measurements of leaf water potential, and flow rate of sap in the stems of young, potted plants. Cycling was also observed in soil-grown, mature orchard trees, as indicated by sap flow in stem and branches. KEY RESULTS: Oscillations in stomatal conductance were caused by the rapid propagation and synchronization of changes in xylem water potential throughout the tree, without rapid changes in atmospheric conditions. CONCLUSIONS: The results show marked stomatal oscillations persisting under natural climatic conditions and underscore the need to discover why this phenomenon is so pronounced in orange trees. PMID- 16478766 TI - Antisense inhibition of Rubisco activase increases Rubisco content and alters the proportion of Rubisco activase in stroma and thylakoids in chloroplasts of rice leaves. AB - BACKGROUND AND AIMS: Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activase (RCA) is a nuclear-encoded chloroplast protein that modifies the conformation of Rubisco, releases inhibitors from active sites, and increases enzymatic activity. It appears to have other functions, e.g. in gibberellin signalling and as a molecular chaperone, which are related to its distribution within the chloroplast. The aim of this research was to resolve uncertainty about the localization of RCA, and to determine whether the distributions of Rubisco and RCA were altered when RCA content was reduced. The monocotyledon, Oryza sativa was used as a model species. METHODS: Gas exchange and Rubisco were measured, and the sub-cellular locations of Rubisco and RCA were determined using immunogold-labelling electron microscopy, in wild-type and antisense rca rice plants. KEY RESULTS: In antisense rca plants, net photosynthetic rate and the initial Rubisco activity decreased much less than RCA content. Immunocytolocalization showed that Rubisco in wild-type and antisense plants was localized in the stroma of chloroplasts. However, the amount of Rubisco in the antisense rca plants was greater than in the wild-type plants. RCA was detected in both the chloroplast stroma and in the thylakoid membranes of wild-type plants. The percentage of RCA labelling in the thylakoid membrane was shown to be substantially decreased, while the fraction in the stroma was increased, by the antisense rca treatment. CONCLUSIONS: From the changes in RCA distribution and alterations in Rubisco activity, RCA in the stroma of the chloroplast probably contributes to the activation of Rubisco, and RCA in thylakoids compensates for the reduction of RCA in the stroma, allowing steady-state photosynthesis to be maintained when RCA is depleted. RCA may also have a second role in protecting membranes against environmental stresses as a chaperone. PMID- 16478767 TI - Sporophyte and gametophyte generations differ in their thermotolerance response in the moss Microbryum. AB - BACKGROUND AND AIMS: Actively growing post-embryonic sporophytes of desert mosses are restricted to the cooler, wetter months. However, most desert mosses have perennial gametophytes. It is hypothesized that these life history patterns are due in part to a reduced thermotolerance for sporophytes relative to gametophytes. METHODS: Gametophytes with attached embryonic sporophytes of Microbryum starckeanum were exposed whilst desiccated to thermal episodes of 35 degrees C (1 hr), 55 degrees C (1 hr), 75 degrees C (1 hr) and 75 degrees C (3 hr), then moistened and allowed to recover for 35 d in a growth chamber. KEY RESULTS: All of the gametophytes survived the thermal exposures and produced protonemata, with the majority also producing shoot buds. Symptoms of gametophytic stress (leaf burning and discoloration of entire shoots) were present in lower frequencies in the 55 degrees C exposure. Sporophyte resumption of growth and maturation to meiosis were significantly negatively affected by thermal treatment. Not a single sporophyte exposed to the two higher thermal treatments (75 degrees C for 1 h and 75 degrees C for 3 h) survived to meiosis, and those sporophytes exposed to 75 degrees C that survived to the post-embryonic phenophase took significantly longer to reach this phase. Furthermore, among the thermal treatments where some capsules reached maturity (35 degrees C and 55 degrees C), maternal shoots that produced a meiotic capsule took longer to regenerate through protonemata than maternal shoots aborting their sporophyte, suggestive of a resource trade-off between generations. CONCLUSIONS: Either (1) the inherent sporophyte thermotolerance is quite low even in this desert moss, and/or (2) a gametophytic thermal stress response controls sporophyte viability. PMID- 16478768 TI - c-jun N-terminal kinase hyperphosphorylates R406W tau at the PHF-1 site during mitosis. AB - Tauopathies such as Alzheimer disease (AD) probably involve a type of phosphorylation imbalance causing the accumulation of abnormally hyperphosphorylated tau in neurons and/or glias. Investigation of R406W tau mutation may provide insight into such abnormal tau hyperphosphorylation, since this mutation causes AD-like dementia and tauopathy in humans and because it has the unique ability to reduce tau phosphorylation in vitro and in cultured cells. Here we show that R406W mutation primarily disrupts tau phosphorylation at Ser404, a priming phosphorylation site of glycogen synthase kinase-3beta (GSK 3beta), thereby reducing subsequent GSK-3beta-mediated phosphorylation at the PHF 1 site (mostly Ser396). In contrast, c-jun N-terminal kinase (JNK) as activated in the mitotic phase directly hyperphosphorylates R406W tau at the PHF-1 site. This was confirmed by PHF-1 hyperphosphorylation of R406W tau in mitotic cells, its association with cytoplasmic JNK activation, and its inhibition by a JNK inhibitor, SP600125. These data unveil the unknown mechanisms of physiological tau phosphorylation at the PHF-1 site and suggest that cytoplasmic JNK activation may play an important role in the abnormal tau hyperphosphorylation associated with R406W tau mutation and in AD. PMID- 16478769 TI - Promoter hypomethylation of Toll-like receptor-2 gene is associated with increased proinflammatory response toward bacterial peptidoglycan in cystic fibrosis bronchial epithelial cells. AB - Cystic fibrosis (CF) is the most common lethal inherited disorder caused by mutation in the gene encoding CF transmembrane regulator (CFTR). The clinical course of CF is characterized by recurrent pulmonary infections and chronic inflammation. Here, we show that toll-like receptor-2 (TLR2) expression and response were strongly enhanced in the human CF bronchial epithelial cell line, CFBE41o-. Treatment of the cells with 5-azacytidine decreased the promoter methylation within TLR2 proximal promoter and increased endogenous expression of TLR2 in non-CF 16HBE14o- cells, suggesting that TLR2 expression is epigenetically regulated by CpG methylation. Moreover, bisulfite sequence analysis revealed that TLR2 promoters were highly demethylated in CFBE41o- cells, implying that decreased methylation of the TLR2 promoter is responsible for CF-related up regulation of TLR2. Finally, stable expression of WT-CFTR in CFBE41o- cells (CFBE41o-/WT-CFTR cells) reduced TLR2 expression and the response to its ligand peptidoglycan (PGN), implying a causal relationship between CFTR dysfunction and TLR2 up-regulation. Consistent with reduced expression of TLR2 in CFBE41o-/WT CFTR cells, CpG methylation was increased in CFBE41o-/WT-CFTR cells. Taken together, our results demonstrate that TLR2 expression is epigenetically up regulated in CF bronchial epithelial cells and suggest that TLR2 overexpression or prolonged activation of TLR2 signaling might be critical in CF pathogenesis. PMID- 16478770 TI - Glycogen branches out: new perspectives on the role of glycogen metabolism in the integration of metabolic pathways. AB - Glycogen is the storage form of carbohydrate for virtually every organism from yeast to primates. Most mammalian tissues store glucose as glycogen, with the major depots located in muscle and liver. The French physiologist Claude Bernard first identified a starch-like substance in liver and muscle and coined the term glycogen, or "sugar former," in the 1850s. During the 150 years since its identification, researchers in the field of glycogen metabolism have made numerous discoveries that are now recognized as significant milestones in biochemistry and cell signaling. Even so, more questions remain, and studies continue to demonstrate the complexity of the regulation of glycogen metabolism. Under classical definitions, the functions of glycogen seem clear: muscle glycogen is degraded to generate ATP during increased energy demand, whereas hepatic glycogen is broken down for release of glucose into the bloodstream to supply other tissues. However, recent findings demonstrate that the roles of glycogen metabolism in energy sensing, integration of metabolic pathways, and coordination of cellular responses to hormonal stimuli are far more complex. PMID- 16478771 TI - Chronic selective glycogen synthase kinase-3 inhibition enhances glucose disposal and muscle insulin action in prediabetic obese Zucker rats. AB - Increasing evidence supports a negative role of glycogen synthase kinase-3 (GSK 3) in regulation of skeletal muscle glucose transport. We assessed the effects of chronic treatment of insulin-resistant, prediabetic obese Zucker (fa/fa) rats with a highly selective GSK-3 inhibitor (CT118637) on glucose tolerance, whole body insulin sensitivity, plasma lipids, skeletal muscle insulin signaling, and in vitro skeletal muscle glucose transport activity. Obese Zucker rats were treated with either vehicle or CT118637 (30 mg/kg body wt) twice per day for 10 days. Fasting plasma insulin and free fatty acid levels were reduced by 14 and 23% (P < 0.05), respectively, in GSK-3 inhibitor-treated animals compared with vehicle-treated controls. The glucose response during an oral glucose tolerance test was reduced by 18% (P < 0.05), and whole body insulin sensitivity was increased by 28% (P < 0.05). In vivo insulin receptor substrate-1 (IRS-1) tyrosine phosphorylation (50%) and IRS-1-associated phosphatidylinositol-3' kinase (79%) relative to fasting plasma insulin levels were significantly elevated (P < 0.05) in plantaris muscles of GSK-3 inhibitor-treated animals. Whereas basal glucose transport in isolated soleus and epitrochlearis muscles was unaffected by chronic GSK-3 treatments, insulin stimulation of glucose transport above basal was significantly enhanced (32-60%, P < 0.05). In summary, chronic treatment of insulin-resistant, prediabetic obese Zucker rats with a specific GSK 3 inhibitor enhances oral glucose tolerance and whole body insulin sensitivity and is associated with an amelioration of dyslipidemia and an improvement in IRS 1-dependent insulin signaling in skeletal muscle. These results provide further evidence that selective targeting of GSK-3 in muscle may be an effective intervention for the treatment of obesity-associated insulin resistance. PMID- 16478772 TI - Liver steatosis coexists with myocardial insulin resistance and coronary dysfunction in patients with type 2 diabetes. AB - Nonalcoholic fatty liver (NAFL) is a common comorbidity in patients with type 2 diabetes and links to the risk of coronary syndromes. The aim was to determine the manifestations of metabolic syndrome in different organs in patients with liver steatosis. We studied 55 type 2 diabetic patients with coronary artery disease using positron emission tomography. Myocardial perfusion was measured with [15O]H2O and myocardial and skeletal muscle glucose uptake with 2-deoxy-2 [18F]fluoro-D-glucose during hyperinsulinemic euglycemia. Liver fat content was determined by magnetic resonance proton spectroscopy. Patients were divided on the basis of their median (8%) into two groups with low (4.6 +/- 2.0%) and high (17.4 +/- 8.0%) liver fat content. The groups were well matched for age, BMI, and fasting plasma glucose. In addition to insulin resistance at the whole body level (P = 0.012) and muscle (P = 0.002), the high liver fat group had lower insulin stimulated myocardial glucose uptake (P = 0.040) and glucose extraction rate (P = 0.0006) compared with the low liver fat group. In multiple regression analysis, liver fat content was the most significant explanatory variable for myocardial insulin resistance. In addition, the high liver fat group had increased concentrations of high sensitivity C-reactive protein, soluble forms of E selectin, vascular adhesion protein-1, and intercellular adhesion molecule-1 (P < 0.05) and lower coronary flow reserve (P = 0.02) compared with the low liver fat group. In conclusion, in patients with type 2 diabetes and coronary artery disease, liver fat content is a novel independent indicator of myocardial insulin resistance and reduced coronary functional capacity. Further studies will reveal the effect of hepatic fat reduction on myocardial metabolism and coronary function. PMID- 16478773 TI - Modulation of muscle protein synthesis by insulin is maintained during neonatal endotoxemia. AB - Sepsis promotes insulin resistance and reduces protein synthesis in skeletal muscle of adults. The effect of sepsis on insulin-stimulated muscle protein synthesis has not been determined in neonates, a highly anabolic population that is uniquely sensitive to insulin. Overnight fasted neonatal pigs were infused for 8 h with endotoxin [lipopolysaccharide (LPS), 0 and 10 mug.kg(-1).h(-1)]. Glucose and amino acids were maintained at fasting levels, insulin was clamped at either fasting or fed (2 or 10 muU/ml) levels, and fractional protein synthesis rates were determined at the end of the infusion. LPS infusion induced a septic-like state, as indicated by a sustained elevation in body temperature, heart rate, and cortisol. At fasting insulin levels, LPS reduced fractional protein synthesis rates in gastrocnemius muscle (-26%) but had no effect on the masseter and heart. By contrast, LPS stimulated liver protein synthesis (+28%). Increasing insulin to fed levels accelerated protein synthesis rates in gastrocnemius (controls by +38%, LPS by +60%), masseter (controls by +50%, LPS by +43%), heart (controls by +34%, LPS by +40%), and diaphragm (controls by +54%, LPS by +29%), and the response to insulin was similar in LPS and controls. Insulin did not alter protein synthesis in liver, kidney, or jejunum in either group. These findings suggest that acute endotoxemia lowers basal fasting muscle protein synthesis in neonates but does not alter the response of protein synthesis to insulin. PMID- 16478774 TI - Infusion of ACTH stimulates expression of adrenal ACTH receptor and steroidogenic acute regulatory protein mRNA in fetal sheep. AB - The late-gestation plasma cortisol surge in the sheep fetus is critical for stimulating organ development and parturition. Increased adrenal responsiveness is one of the key reasons for the surge; however, the underlying mechanisms are not fully understood. Our recent studies suggest that ACTH-mediated increased expression of ACTH receptor (ACTH-R) and steroid acute regulatory protein (StAR) may play a role in enhancing responsiveness. Hence, we examined effects of ACTH infusion in fetal sheep on mRNA expression of these two mediators of adrenal responsiveness and assessed the functional consequences of this treatment in vitro. Fetuses of approximately 118 and 138 days of gestational age (dGA) were infused with ACTH-(1-24) for 24 h. Controls received saline infusion. Arterial blood was sampled throughout the infusion. Adrenals were isolated and analyzed for ACTH-R and StAR mRNA, or cells were cultured for 48 h. Cells were stimulated with ACTH, and medium was collected for cortisol measurement. Fetal plasma ACTH and cortisol concentrations increased over the infusion period in both groups. ACTH-R mRNA levels were significantly higher in ACTH-infused fetuses in both the 118 and 138 dGA groups. StAR mRNA increased significantly in both the 118 and 138 dGA groups. Adrenal cells from ACTH-infused fetuses were significantly more responsive to ACTH stimulation in terms of cortisol secretion than those from saline-infused controls. These findings demonstrate that increases in circulating ACTH levels promote increased expression of ACTH-R and StAR mRNA and are coupled to heightened adrenal responsiveness. PMID- 16478775 TI - Impact of incretin hormones on beta-cell function in subjects with normal or impaired glucose tolerance. AB - The mechanisms by which the enteroinsular axis influences beta-cell function have not been investigated in detail. We performed oral and isoglycemic intravenous (IV) glucose administration in subjects with normal (NGT; n = 11) or impaired glucose tolerance (IGT; n = 10), using C-peptide deconvolution to calculate insulin secretion rates and mathematical modeling to quantitate beta-cell function. The incretin effect was taken to be the ratio of oral to IV responses. In NGT, incretin-mediated insulin release [oral glucose tolerance test (OGTT)/IV ratio = 1.59 +/- 0.18, P = 0.004] amounted to 18 +/- 2 nmol/m(2) (32 +/- 4% of oral response), and its time course matched that of total insulin secretion. The beta-cell glucose sensitivity (OGTT/IV ratio = 1.52 +/- 0.26, P = 0.02), rate sensitivity (response to glucose rate of change, OGTT/IV ratio = 2.22 +/- 0.37, P = 0.06), and glucose-independent potentiation were markedly higher with oral than IV glucose. In IGT, beta-cell glucose sensitivity (75 +/- 14 vs. 156 +/- 28 pmol.min(-1).m(-2).mM(-1) of NGT, P = 0.01) and potentiation were impaired on the OGTT. The incretin effect was not significantly different from NGT in terms of plasma glucagon-like peptide 1 and glucose-dependent insulinotropic polypeptide responses, total insulin secretion, and enhancement of beta-cell glucose sensitivity (OGTT/IV ratio = 1.73 +/- 0.24, P = NS vs. NGT). However, the time courses of incretin-mediated insulin secretion and potentiation were altered, with a predominance of glucose-induced vs. incretin-mediated stimulation. We conclude that, under physiological circumstances, incretin-mediated stimulation of insulin secretion results from an enhancement of all dynamic aspects of beta cell function, particularly beta-cell glucose sensitivity. In IGT, beta-cell function is inherently impaired, whereas the incretin effect is only partially affected. PMID- 16478777 TI - Characterization of L-arginine transport in adrenal cells: effect of ACTH. AB - Nitric oxide synthesis depends on the availability of its precursor L-arginine, which could be regulated by the presence of a specific uptake system. In the present report, the characterization of the L-arginine transport system in mouse adrenal Y1 cells was performed. L-arginine transport was mediated by the cationic/neutral amino acid transport system y+L and the cationic amino acid transporter (CAT) y+ in Y1 cells. These Na+-independent transporters were identified by their selectivity for neutral amino acids in both the presence and absence of Na+ and by the effect of N-ethylmaleimide. Transport data correlated to expression of genes encoding for CAT-1, CAT-2, CD-98, and y+LAT-2. A similar expression profile was detected in rat adrenal zona fasciculata. In addition, cationic amino acid uptake in Y1 cells was upregulated by ACTH and/or cAMP with a concomitant increase in nitric oxide (NO) production. PMID- 16478776 TI - Expression of TPO and ThOXs in human thyrocytes is downregulated by IL-1alpha/IFN gamma, an effect partially mediated by nitric oxide. AB - Morphological and functional alterations in Hashimoto's thyroiditis (HT) are predominantly mediated by Th1 cytokines through apoptotic cell death. This ultimate step could be preceded by functional injuries in thyroid hormone synthesis. The action of two Th1 cytokines (IL-1alpha/IFN-gamma) on thyroperoxidase (TPO) and thyroid oxidase (ThOXs) expression was tested in human thyrocytes isolated from normal tissues, Graves' disease (GD) tissues, and autonomous toxic nodules. There was no evidence of cell death. Nitric oxide (NO) release was induced by cytokines but was absent when NG-nitro-L-arginine methyl ester (L-NAME) was coincubated. When thyrotropin (TSH)-incubated normal and GD thyrocytes were treated with IL-1alpha/IFN-gamma, TPO and ThOXs protein and mRNA expression dropped, a decrease partially prevented by L-NAME, suggesting that NO acts as a mediator of Th1 effects. In thyrocytes from autonomous toxic nodules, the high level of TPO and ThOXs protein expression was not influenced by TSH or by cytokines, a finding partially reproduced when normal thyrocytes were treated with increasing concentrations of TSH. In conclusion, incubation of normal or GD thyrocytes with Th1 cytokines induces a significant reduction in TSH-increased expression of both TPO and ThOXs, an effect partially mediated by NO. The thyroid cell function can therefore be severely affected in HT, even when cells remain viable. In autonomous toxic nodules, cells become partially insensitive to exogenous Th1 cytokines. PMID- 16478778 TI - Evaluation of nonlinear regression approaches to estimation of insulin sensitivity by the minimal model with reference to Bayesian hierarchical analysis. AB - Minimal model analysis of intravenous glucose tolerance test (IVGTT) glucose and insulin concentrations offers a validated approach to measuring insulin sensitivity, but model identification is not always successful. Improvements may be achieved by using alternative settings in the modeling process, although results may differ according to setting, and care must be exercised in combining results. IVGTT data (12 samples, regular test) from 533 men without diabetes was modeled by the traditional nonlinear regression (NLR) approach, using five different permutations of settings. Results were evaluated with reference to the more robust Bayesian hierarchical (BH) approach to model identification and to the proportion of variance they explained in known correlates of insulin sensitivity (age, BMI, blood pressure, fasting glucose and insulin, serum triglyceride, HDL cholesterol, and uric acid concentration). BH analysis was successful in all cases. With NLR analysis, between 17 and 35 IVGTTs were associated with parameter coefficients of variation (PCVs) for minimal model parameters S(I) (insulin sensitivity) and S(G) (glucose effectiveness) of >100%. Systematic use of each different approach in combination reduced this number to five. Mean (interquartile range) S(I)(NLR) was then 3.14 (2.29-4.63) min(-1).mU( 1).l x 10(-4) and 2.56 (1.74-3.83) min(-1).mU(-1).l x 10(-4) for S(I)(BH) (correlation 0.86, P < 0.0001). S(I)(NLR) explained, on average, 10.6% of the variance in known correlates of insulin sensitivity, whereas S(I)(BH) explained 8.5%. In a large body of data, which BH analysis demonstrated could be fully identified, use of alternative modeling settings in NLR analysis could substantially reduce the number of analyses with PCVs >100%. S(I)(NLR) compared favorably with S(I)(BH) in the proportion of variance explained in known correlates of insulin sensitivity. PMID- 16478779 TI - Elevated resistin levels in cirrhosis are associated with the proinflammatory state and altered hepatic glucose metabolism but not with insulin resistance. AB - The adipokine resistin has been implicated in obesity and insulin resistance. Liver cirrhosis is associated with decreased body fat mass and insulin resistance. We determined plasma resistin levels in 57 patients with cirrhosis, 13 after liver transplantation, and 30 controls and correlated these with hemodynamic as well as hepatic and systemic metabolic parameters. Patients with cirrhosis had, dependent on the clinical stage, an overall 86% increase in resistin levels (P < 0.001) with hepatic venous resistin being higher than arterial levels (P < 0.001). Circulating resistin was significantly correlated with plasma TNF-alpha levels (r = 0.62, P < 0.001). No correlation was observed between resistin and hepatic hemodynamics, body fat mass, systemic energy metabolism, and the degree of insulin resistance. However, plasma resistin in cirrhosis was negatively associated with hepatic glucose production (r = -0.47, P < 0.01) and positively with circulating free fatty acids (FFA; r = 0.40, P < 0.01) and ketone bodies (r = 0.48, P < 0.001) as well as hepatic ketone body production (r = 0.40, P < 0.01). After liver transplantation, plasma resistin levels remained unchanged, whereas insulin resistance was significantly improved (P < 0.01). These data provide novel insights into the role of resistin in the pathophysiological background of a catabolic disease in humans and also indicate that resistin inhibition may not represent a suitable therapeutic strategy for the treatment of insulin resistance and diabetes in patients with liver cirrhosis. PMID- 16478780 TI - Metformin counters the insulin-induced suppression of fatty acid oxidation and stimulation of triacylglycerol storage in rodent skeletal muscle. AB - The present study examined the acute effects of metformin on fatty acid (FA) metabolism in oxidative soleus (SOL) and glycolytic epitrochlearis (EPT) rodent muscle. SOL and EPT were incubated for either 30 or 180 min in the absence or presence of 2 mM metformin and with or without insulin (10 mU/ml). Metformin did not alter basal FA metabolism but countered the effects of insulin on FA oxidation and incorporation into triacylglyerol (TAG). Specifically, metformin prevented the insulin-induced suppression of FA oxidation in SOL but did not alter FA incorporation into lipid pools. In contrast, in EPT metformin blunted the incorporation of FA into TAG when insulin was present but did not alter FA oxidation. In SOL, metformin resulted in a 50% increase in AMP-activated protein kinase alpha2 activity and prevented the insulin-induced increase in malonyl-CoA content. In both fiber types, basal and insulin-stimulated glucose oxidation were not significantly altered by metformin. All effects were similar regardless of whether they were measured after 30 or 180 min. Because increased muscle lipid storage and impaired FA oxidation have been associated with insulin resistance in this tissue, the ability of metformin to reverse these abnormalities in muscle FA metabolism may be a part of the mechanism by which metformin improves glucose clearance and insulin sensitivity. The present data also suggest that increased glucose clearance is not due to its enhanced subsequent oxidation. Additional studies are warranted to determine whether chronic metformin treatment has similar effects on muscle FA metabolism. PMID- 16478781 TI - Localized accumulation of angiotensin II and production of angiotensin-(1-7) in rat luteal cells and effects on steroidogenesis. AB - These studies aim to investigate subcellular distribution of angiotensin II (ANG II) in rat luteal cells, identify other bioactive angiotensin peptides, and investigate a role for angiotensin peptides in luteal steroidogenesis. Confocal microscopy showed ANG II distributed within the cytoplasm and nuclei of luteal cells. HPLC analysis showed peaks that eluted with the same retention times as ANG-(1-7), ANG II, and ANG III. Their relative concentrations were ANG II >or= ANG-(1-7) > ANG III, and accumulation was modulated by quinapril, an inhibitor of angiotensin-converting enzyme (ACE), Z-proprolinal (ZPP), an inhibitor of prolyl endopeptidase (PEP), and parachloromercurylsulfonic acid (PCMS), an inhibitor of sulfhydryl protease. Phenylmethylsulfonyl fluoride (PMSF), a serine protease inhibitor, did not affect peptide accumulation. Quinapril, ZPP, PCMS, and PMSF, as well as losartan and PD-123319, the angiotensin receptor type 1 (AT1) and type 2 (AT2) receptor antagonists, were used in progesterone production studies. ZPP significantly reduced luteinizing hormone (LH)-dependent progesterone production (P < 0.05). Quinapril plus ZPP had a greater inhibitory effect on LH-stimulated progesterone than either inhibitor alone, but this was not reversed by exogenous ANG II or ANG-(1-7). Both PCMS and PMSF acutely blocked LH-stimulated progesterone, and PCMS blocked LH-sensitive cAMP accumulation. Losartan inhibited progesterone production in permeabilized but not intact luteal cells and was reversed by ANG II. PD-123319 had no significant effect on luteal progesterone production in either intact or permeabilized cells. These data suggest that steroidogenesis may be modulated by angiotensin peptides that act in part through intracellular AT1 receptors. PMID- 16478782 TI - Regulation of dishevelled and beta-catenin in rat skeletal muscle: an alternative exercise-induced GSK-3beta signaling pathway. AB - beta-catenin is a multifunctional protein involved in cell-cell adhesion and the Wnt signaling pathway. beta-Catenin is activated upon its dephosphorylation, an event triggered by Dishevelled (Dvl)-mediated phosphorylation and deactivation of glycogen synthase kinase-3beta (GSK-3beta). In skeletal muscle, both insulin and exercise decrease GSK-3beta activity, and we tested the hypothesis that these two stimuli regulate beta-catenin. Immunoblotting demonstrated that Dvl, Axin, GSK 3beta, and beta-catenin proteins are expressed in rat red and white gastrocnemius muscles. Treadmill running exercise in vivo significantly decreased beta-catenin phosphorylation in both muscle types, with complete dephosphorylation being elicited by maximal exercise. beta-Catenin dephosphorylation was intensity dependent, as dephosphorylation was highly correlated with muscle glycogen depletion during exercise (r(2) = 0.84, P < 0.001). beta-Catenin dephosphorylation was accompanied by increases in GSK-3beta Ser(9) phosphorylation and Dvl-GSK-3beta association. In contrast to exercise, maximal insulin treatment (1 U/kg body wt) had no effect on skeletal muscle beta-catenin phosphorylation or Dvl-GSK-3beta interaction. In conclusion, exercise in vivo, but not insulin, increases the association between Dvl and GSK-3beta in skeletal muscle, an event paralleled by beta-catenin dephosphorylation. PMID- 16478783 TI - The polarity-establishment component Bem1p interacts with the exocyst complex through the Sec15p subunit. AB - Spatial regulation of the secretory machinery is essential for the formation of a new bud in Saccharomyces cerevisiae. Yet, the mechanisms underlying cross-talk between the secretory and the cell-polarity-establishment machineries have not been fully elucidated. Here, we report that Sec15p, a subunit of the exocyst complex, might provide one line of communication. Not only is Sec15p an effector of the rab protein Sec4p, the master regulator of post-Golgi trafficking, but it also interacts with components of the polarity-establishment machinery. We have demonstrated a direct physical interaction between Sec15p and Bem1p, a protein involved in the Cdc42p-mediated polarity-establishment pathway, confirming a prior two-hybrid study. When this interaction is compromised, as in the case of cells lacking the N-terminal 138 residues of Bem1p, including the first Src homology 3 (SH3) domain, the localization of green fluorescent protein (GFP) tagged Sec15 is affected, especially in the early stage of bud growth. In addition, Sec15-1p, which is defective in Bem1p binding, mislocalizes along with Sec8p, another exocyst subunit. Overall, our evidence suggests that the interaction of Sec15p with Bem1p is important for Sec15p localization at the early stage of bud growth and, through this interaction, Sec15p might play a crucial role in integrating the signals between Sec4p and the components of the early-polarity-establishment machinery. This, in turn, helps to coordinate the secretory pathway and polarized bud growth. PMID- 16478784 TI - Ca2+ stimulation of adenylyl cyclase generates dynamic oscillations in cyclic AMP. AB - The spatial and temporal complexity of Ca2+ signalling is central to the regulation of a diverse range of cellular processes. The decoding of dynamic Ca2+ signals is, in part, mediated by the ability of Ca2+ to regulate other second messengers, including cyclic AMP (cAMP). A number of kinetic models (including our own) predict that interdependent Ca2+ and cAMP oscillations can be generated. A previous study in Xenopus neurons illustrated prolonged, low-frequency cAMP oscillations during bursts of Ca2+ transients. However, the detection of more dynamic Ca2+ driven changes in cAMP has, until recently, been limited by the availability of suitable cAMP probes with high temporal resolution. We have used a newly developed FRET-based cAMP indicator comprised of the cAMP binding domain of Epac-1 to examine interplay between Ca2+ and cAMP dynamics. This probe was recently used in excitable cells to reveal an inverse relationship between cAMP and Ca2+ oscillations as a consequence of Ca2+-dependent activation of phosphodiesterase 1 (PDE1). Here, we have used human embryonic kidney (HEK293) cells expressing the type 8 adenylyl cyclase (AC8) to examine whether dynamic Ca2+ changes can mediate phasic cAMP oscillations as a consequence of Ca2+ stimulated AC activity. During artificial or agonist-induced Ca2+ oscillations we detected fast, periodic changes in cAMP that depended upon Ca2+ stimulation of AC8 with subsequent PKA-mediated phosphodiesterase 4 (PDE4) activity. Carbachol (10 microM) evoked cAMP transients with a peak frequency of approximately 3 minute(-1), demonstrating phasic oscillations in cAMP and Ca2+ in response to physiological stimuli. Furthermore, by imposing a range of Ca2+-oscillation frequencies, we demonstrate that AC8 acts as a low-pass filter for high-frequency Ca2+ events, enhancing the regulatory options available to this signalling pathway. PMID- 16478785 TI - Acidic clusters target transmembrane proteins to the contractile vacuole in Dictyostelium cells. AB - The mechanisms responsible for the targeting of transmembrane integral proteins to the contractile vacuole (CV) network in Dictyostelium discoideum are unknown. Here we show that the transfer of the cytoplasmic domain of a CV-resident protein (Rh50) to a reporter transmembrane protein (CsA) is sufficient to address the chimera (CsA-Rh50) to the CV. We identified two clusters of acidic residues responsible for this targeting, and these motifs interacted with the gamma adaptin AP-1 subunit in a yeast protein-protein interaction assay. For the first time we report the existence of an indirect transport pathway from the plasma membrane to the CV via endosomes. Upon internalization, the small fraction of CsA Rh50 present at the cell surface was first concentrated in endosomes distinct from early and late p80-positive endosomes and then slowly transported to the CV. Together our results suggest the existence of an AP-1-dependent selective transport to the contractile vacuole in Dictyostelium. PMID- 16478786 TI - Silencing of the hydra serine protease inhibitor Kazal1 gene mimics the human SPINK1 pancreatic phenotype. AB - In hydra, the endodermal epithelial cells carry out the digestive function together with the gland cells that produce zymogens and express the evolutionarily conserved gene Kazal1. To assess the hydra Kazal1 function, we silenced gene expression through double-stranded RNA feeding. A progressive Kazal1 silencing affected homeostatic conditions as evidenced by the low budding rate and the induced animal death. Concomitantly, a dramatic disorganization followed by a massive death of gland cells was observed, whereas the cytoplasm of digestive cells became highly vacuolated. The presence of mitochondria and late endosomes within those vacuoles assigned them as autophagosomes. The enhanced Kazal1 expression in regenerating tips was strongly diminished in Kazal1(-) hydra, and the amputation stress led to an immediate disorganization of the gland cells, vacuolization of the digestive cells and death after prolonged silencing. This first cellular phenotype resulting from a gene knock-down in cnidarians suggests that the Kazal1 serine-protease-inhibitor activity is required to prevent excessive autophagy in intact hydra and to exert a cytoprotective function to survive the amputation stress. Interestingly, these functions parallel the pancreatic autophagy phenotype observed upon mutation within the Kazal domain of the SPINK1 and SPINK3 genes in human and mice, respectively. PMID- 16478787 TI - Characterization of a neural-specific splicing form of the human neuregulin 3 gene involved in oligodendrocyte survival. AB - Neuregulins are a family of genes involved in key aspects of neural biology. Neuregulins 1, 2 and 3 (NRG1, NRG2 and NRG3) are expressed in the mammalian nervous system. It is well established that NRG1, with fifteen different splicing forms, is central for brain development and function. However, the biological relevance of NRG2 and NRG3 remains elusive. Here, we report the identification of a new isoform of NRG3 that is specifically expressed in the human embryonic central nervous system. Sequence alignment with the human genome suggests that this transcript is produced by alternative promoter usage. The encoded polypeptide is a type-I-glycosylated plasma membrane protein, which is shed into the extracellular space where it activates erbB4, a pivotal receptor for brain development. In addition, we show that the protein has a signal sequence that is cleaved after membrane insertion. Proteasome inhibition with Lactacystin enhances the expression of the protein, whereas impairment of ubiquitylation in the conditional mutant cell line ts20 protects the protein from degradation. These observations imply that the ubiquitin/proteasome pathway regulates biogenesis of the protein. We also show that recombinant neuregulin 3 acts as an oligodendrocyte survival factor by activating the phosphoinositide 3-kinase signalling pathway. Therefore, we report a new post-translationally regulated isoform of neuregulin 3 expressed in the developing human central nervous system with a role in oligodendrocyte survival. PMID- 16478788 TI - Molecular mapping of tyrosine-phosphorylated proteins in focal adhesions using fluorescence resonance energy transfer. AB - Microscopy-based fluorescence resonance energy transfer (FRET) provides an opportunity to monitor molecular processes in the natural environment in live cells. Here we studied molecular interactions and tyrosine phosphorylation of paxillin, Crk-associated substrate (CAS), and focal adhesion kinase (FAK) in focal adhesions. For that purpose, these focal adhesion phosphoproteins, fused to cyan or yellow fluorescent proteins (CFP or YFP) were expressed in cultured fibroblasts. To assess the dynamics of tyrosine phosphorylation we used YFP- or CFP-tagged SH2 domain of pp60(src) (dSH2), which specifically binds to phosphotyrosine residues. FRET measurements, combined with immunolabeling with phosphospecific antibodies revealed that FAK, CAS and paxillin are tyrosine phosphorylated in early matrix adhesions and that FAK is in FRET proximity to CAS and paxillin in focal complexes and focal adhesions. Data suggest that paxillin incorporation into nascent focal complexes precedes its tyrosine phosphorylation, which then gradually increases. In cells treated with Rho-kinase inhibitors or expressing constitutively active Rac, focal complexes showed similar levels of paxillin tyrosine phosphorylation as seen in mature focal adhesions. Dynamic FRET based examination indicated that paxillin phosphorylation occurs in specific areas (hotspots) within focal adhesions, whereas FAK phosphorylation is broadly distributed. PMID- 16478789 TI - PKBalpha is required for adipose differentiation of mouse embryonic fibroblasts. AB - Protein kinase Balpha (PKBalpha) is a key regulator of metabolism, proliferation and differentiation. We have explored the role of PKBalpha in adipogenesis using wild-type and PKBalpha-knockout mouse embryonic fibroblasts (MEFs) and show that lack of PKBalpha prevents MEF differentiation into adipocytes. Expression of ectopic PKBalpha in PKBalpha-deficient cells restores adipogenesis. We identified 80 genes whose expression was upregulated in wild-type MEFs during adipogenesis but whose expression was significantly reduced in PKBalpha-deficient MEFs under the same conditions. Significantly, the regulator of adipogenesis Kruppel-like transcription factor 15 gene expression was downregulated in PKBalpha-deficient MEFs but could be restored by expressing an active PKBalpha in the deficient cells. The level of lipocalin 2, renin 1 and receptor-activity-modifying protein 3 genes expressed by adipose cells was also decreased in PKBalpha-deficient MEFs, and are inhibited by LY294002 treatment during early adipocyte differentiation of 3T3-L1 cells. The results underscore an essential role for PKBalpha in the transcriptional program required for adipogenesis. PMID- 16478791 TI - APC inhibits ERK pathway activation and cellular proliferation induced by RAS. AB - Inactivating mutations in the adenomatous polyposis coli gene (APC), and activating mutations in RAS, occur in a majority of colorectal carcinomas. However, the relationship between these changes and tumorigenesis is poorly understood. RAS-induced activation of the ERK pathway was reduced by overexpressing APC in DLD-1 colorectal cancer cells. ERK activity was increased by Cre-virus-induced Apc knockout in primary Apc(flox/flox) mouse embryonic fibroblasts, indicating that APC inhibits ERK activity. ERK activity was increased by overexpression and decreased by knock down of beta-catenin. The activation of Raf1, MEK and ERK kinases by beta-catenin was reduced by co expression of APC. These results indicate that APC inhibits the ERK pathway by an action on beta-catenin. RAS-induced activation of the ERK pathway was reduced by the dominant negative form of TCF4, indicating that the ERK pathway regulation by APC/beta-catenin signaling is, at least, partly caused by effects on beta catenin/TCF4-mediated gene expression. The GTP loading and the protein level of mutated RAS were decreased in cells with reduced ERK activity as a result of APC overexpression, indicating that APC regulates RAS-induced ERK activation at least partly by reduction of the RAS protein level. APC regulates cellular proliferation and transformation induced by activation of both RAS and beta catenin signaling. PMID- 16478790 TI - A role for Sec8 in oligodendrocyte morphological differentiation. AB - In the central nervous system, oligodendrocytes synthesize vast amounts of myelin, a multilamellar membrane wrapped around axons that dramatically enhances nerve transmission. A complex apparatus appears to coordinate trafficking of proteins and lipids during myelin synthesis, but the molecular interactions involved are not well understood. We demonstrate that oligodendrocytes express several key molecules necessary for the targeting of transport vesicles to areas of rapid membrane growth, including the exocyst components Sec8 and Sec6 and the multidomain scaffolding proteins CASK and Mint1. Sec8 overexpression significantly promotes oligodendrocyte morphological differentiation and myelin like membrane formation in vitro; conversely, siRNA-mediated interference with Sec8 expression inhibits this process, and anti-Sec8 antibody induces a reduction in oligodendrocyte areas. In addition, Sec8 colocalizes, coimmunoprecipitates and cofractionates with the major myelin protein OSP/Claudin11 and with CASK in oligodendrocytes. These results suggest that Sec8 plays a central role in oligodendrocyte membrane formation by regulating the recruitment of vesicles that transport myelin proteins such as OSP/Claudin11 to sites of membrane growth. PMID- 16478792 TI - Possible involvement of hyperlipidemia in increasing risk of colorectal tumor development in human familial adenomatous polyposis. AB - BACKGROUND: Familial adenomatous polyposis (FAP) results from germline adenomatous polyposis coli (APC) gene mutations and many affected patients die from colorectal cancers which arise from colorectal polyps. We previously reported that two strains of Apc gene-deficient mice developing multiple intestinal polyps exhibit a hyperlipidemic state. The triglyceride (TG) levels were approximately 10-fold higher than the levels observed in wild-type mice. METHODS: To examine whether a positive relationship might exist between hyperlipidemia and colorectal tumor development in FAP patients, as with Apc gene deficient mice, a pilot experiment was performed using readily available clinical data such as ages, serum lipid levels, number of colorectal polyps and cancer development in 28 FAP patients from the National Cancer Center Hospital, Japan. RESULTS: The overall prevalence of hyperlipidemia in FAP cases was 58%. Average TG levels in the 40-60 year age groups of FAP patients were > or =150 mg/dl (the defined threshold level of hyperlipidemia). Moreover, there was a tendency for higher serum TG levels in patients who developed colorectal cancer, as compared with those without colorectal cancer. CONCLUSIONS: These results show that a hyperlipidemic state occurs in FAP patients. Although it is weaker than that in Apc gene-deficient mice, it may be linked to colon tumor development. These data warrant further studies for wider populations of FAP patients. PMID- 16478793 TI - Extended right hemihepatectomy as a salvage operation for recurrent bile duct cancer 3 years after pancreatoduodenectomy. AB - Salvage surgery for recurrent bile duct cancer is generally impractical due to local invasion of surrounding major vascular structures or distant metastases. We describe a case of a relapsed tumor in the right hepatic duct 3 years after pancreatoduodenectomy for middle to distal bile duct cancer. The recurrent tumor, measuring 25 x 12 x 12 mm, was mostly confined within the right hepatic duct. It displayed an intraductal superficial extension rather than transmural invasive growth to the hepatic hilum. An extended right hemihepatectomy was successfully performed with a histologically negative margin. The patient is currently doing well without any signs of local recurrence or distant metastasis 8 months after the second operation. Precise pathological examination revealed that the lesion had originated from multicentric foci in the right hepatic duct, not as a result of anastomotic recurrence. These results raised the consideration of a potentially more indolent subgroup of bile duct cancer. This is a detailed report of a successfully resected recurrent bile duct cancer, for which the patient underwent major hepatectomy as a salvage procedure after pancreatoduodenectomy for the primary tumor. An aggressive surgical approach will be a rational treatment of choice for recurrent disease when metachronous multicentric tumor development in the bile duct is suspected and curative resection can be safely performed. PMID- 16478794 TI - MDR1 polymorphisms predict the response to etoposide-cisplatin combination chemotherapy in small cell lung cancer. AB - BACKGROUND: The MDR1 gene encodes P-glycoprotein (PGP), which plays an important role in mediating multidrug resistance to chemotherapeutic agents. Polymorphisms in the MDR1 gene may have an impact on the expression and function of PGP, thereby influencing the response to chemotherapy. METHODS: We investigated the potential association of MDR1 polymorphisms (2677G>T at exon 21 and 3435C>T at exon 26) and their haplotypes with chemotherapy response in 54 small cell lung cancer (SCLC) patients who received a combination chemotherapy of etoposide cisplatin. RESULTS: The 3435 CC genotype was associated with a significantly better chemotherapy response compared with the combined 3435 CT and TT genotype (P = 0.025). The 2677 GG genotype was also associated with a better chemotherapy response compared with the combined 2677 GT and TT genotype, although it was not statistically significant. Consistent with the results of genotyping analyses, patients harboring the 2677G-3435C haplotype had a statistically significant better response to chemotherapy compared with those with the other haplotypes combined (P = 0.015). CONCLUSIONS: Our findings suggest that the MDR1 2677G>T and 3435C>T polymorphisms can be used for predicting treatment response to etoposide cisplatin chemotherapy in SCLC patients. PMID- 16478795 TI - Rapid-access chest pain clinics and the traditional cardiology out-patient clinic. AB - BACKGROUND: The National Service Framework for coronary heart disease recommends rapid-access chest pain clinics (RACPCs) for cardiological assessment of new onset chest pain within 2 weeks of referral. AIM: To measure the extent to which an RACPC successfully substituted for an out-patient cardiology clinic (OPCC) at a general hospital, in assessing new-onset chest pain referrals. METHODS: Prospective measurement of attendance and waiting times for consecutive patients at the RACPC and OPCC, and multivariate analysis of factors associated with referral for angiography. RESULTS: From September 2002 to August 2004, 1382 patients with chest pain attended the RACPC, and 228 patients, the OPCC. All RACPC patients were seen within 24 h of referral, except those referred on Friday afternoons, or the day before national holidays. The mean +/- SD waiting time for OPCC appointments was 97 +/- 43 days. Of 208 OPCC patients, 30 (14%) fulfilled the RACPC referral criterion of recent onset chest pain (<4 weeks duration) vs. 926/1382 (67%) RACPC patients. Thus the RACPC substituted for the OPCC in 926/956 (97%) new chest pain referrals. Patients from the OPCC were 3.82 (95%CI 1.85 7.90) more likely to be referred for a coronary angiogram. compared to those attending the RACPC. DISCUSSION: The RACPC has provided efficient and effective substitution for the OPCC in the assessment of new chest pain referrals according to pre-defined referral criteria. Broadening the referral criterion of the RACPC to patients with chest pain of >4 weeks duration would result in more referrals. PMID- 16478796 TI - Central adiposity, regional fat distribution, and the risk of cholecystectomy in women. AB - PURPOSE: Whether central adiposity contributes independently of total adiposity to the risk for gall stones is inconclusive. We examined prospectively indicators of central adiposity in relation to the occurrence of gall stone disease. METHODS: We evaluated the relationship between abdominal circumference and waist to hip ratio and risk of cholecystectomy in a cohort of women who had no history of gall stone disease. As part of the Nurses' Health Study, the women reported on questionnaires their weights, heights, and waist and hip circumferences, and the occurrence of cholecystectomy. A total of 42,312 women, aged 39-66 years in 1986, who were free of prior gall stone disease, provided complete waist and hip circumference measurements in 1986. RESULTS: We documented 3197 cases of cholecystectomy during 514,283 person years of follow up. After adjusting simultaneously for regional (waist circumference or waist to hip ratio) and total adiposity (body mass index) measures as well as for other risk factors of gall stone disease, women with a height adjusted waist circumference of 36 inches or larger had a relative risk (RR) of 1.96 (95% confidence interval (CI) 1.53-2.51; p trend < 0.0001) compared with women with a height adjusted waist circumference of less than 26 inches. Waist to hip ratio was directly associated with the risk, with an RR of 1.39 (95% CI 1.16-1.66; p trend < 0.0001) for women with a waist to hip ratio of 0.86 or higher compared with women with a waist to hip ratio of less than 0.70. CONCLUSION: Abdominal circumference and waist to hip ratio were associated with an increased risk of cholecystectomy, independently of body mass index in women. PMID- 16478797 TI - Heterogeneity of metallo-beta-lactamases in clinical isolates of Chryseobacterium meningosepticum from Hangzhou, China. AB - OBJECTIVES: To determine the distribution and heterogeneity of metallo-beta lactamases (MBLs) responsible for imipenem resistance in Chryseobacterium meningosepticum. METHODS: Clinical C. meningosepticum isolates (n = 170) were collected from hospitals in Hangzhou, China. Production of MBLs was investigated by determination of imipenem MICs, and by using both a three-dimensional test and a 2-mercaptopropionic acid inhibitory test. Genes encoding BlaB and GOB MBLs were amplified by PCR, sequenced and compared with genes in GenBank. RESULTS: More than 95% of the 170 isolates showed high (MIC > 16 mg/L) or intermediate resistance to imipenem, but only 94 isolates (55%) were shown phenotypically to produce MBLs (imipenem MIC range, 8-256 mg/L), with MBL genes detected in 93 of these. Among them, 83 isolates had blaB alleles and 65 isolates had bla(GOB) alleles; 38 isolates possessed one MBL gene and 55 isolates contained two genes. The major blaB alleles encoded BlaB-2, -3 and -11, while the major bla(GOB) alleles encoded GOB-2, -4, -8 and -10. MBLs or their genes were not detected in 76 (45%) isolates, including many that were highly resistant to imipenem. CONCLUSIONS: High levels and rates of imipenem resistance in C. meningosepticum from Hangzhou often result from the presence of heterogeneous BlaB and/or GOB MBLs, although undefined carbapenem resistance mechanisms also exist. Susceptibility testing and screening for MBLs should be conducted in order to inform effective treatment for C. meningosepticum infections. PMID- 16478799 TI - Commentary: games people play--birthweight. PMID- 16478798 TI - Nuclear envelope dystrophies show a transcriptional fingerprint suggesting disruption of Rb-MyoD pathways in muscle regeneration. AB - Mutations of lamin A/C (LMNA) cause a wide range of human disorders, including progeria, lipodystrophy, neuropathies and autosomal dominant Emery-Dreifuss muscular dystrophy (EDMD). EDMD is also caused by X-linked recessive loss-of function mutations of emerin, another component of the inner nuclear lamina that directly interacts with LMNA. One model for disease pathogenesis of LMNA and emerin mutations is cell-specific perturbations of the mRNA transcriptome in terminally differentiated cells. To test this model, we studied 125 human muscle biopsies from 13 diagnostic groups (125 U133A, 125 U133B microarrays), including EDMD patients with LMNA and emerin mutations. A Visual and Statistical Data Analyzer (VISDA) algorithm was used to statistically model cluster hierarchy, resulting in a tree of phenotypic classifications. Validations of the diagnostic tree included permutations of U133A and U133B arrays, and use of two probe set algorithms (MAS5.0 and MBEI). This showed that the two nuclear envelope defects (EDMD LMNA, EDMD emerin) were highly related disorders and were also related to fascioscapulohumeral muscular dystrophy (FSHD). FSHD has recently been hypothesized to involve abnormal interactions of chromatin with the nuclear envelope. To identify disease-specific transcripts for EDMD, we applied a leave one-out (LOO) cross-validation approach using LMNA patient muscle as a test data set, with reverse transcription-polymerase chain reaction (RT-PCR) validations in both LMNA and emerin patient muscle. A high proportion of top-ranked and validated transcripts were components of the same transcriptional regulatory pathway involving Rb1 and MyoD during muscle regeneration (CRI-1, CREBBP, Nap1L1, ECREBBP/p300), where each was specifically upregulated in EDMD. Using a muscle regeneration time series (27 time points) we develop a transcriptional model for downstream consequences of LMNA and emerin mutations. We propose that key interactions between the nuclear envelope and Rb and MyoD fail in EDMD at the point of myoblast exit from the cell cycle, leading to poorly coordinated phosphorylation and acetylation steps. Our data is consistent with mutations of nuclear lamina components leading to destabilization of the transcriptome in differentiated cells. PMID- 16478800 TI - Advancing glycomics: implementation strategies at the consortium for functional glycomics. AB - Glycomics-an integrated approach to study structure-function relationships of complex carbohydrates (or glycans)-is an emerging field in this age of post genomics. Realizing the importance of glycomics, many large scale research initiatives have been established to generate novel resources and technologies to advance glycomics. These initiatives are generating and cataloging diverse data sets necessitating the development of bioinformatic platforms to acquire, integrate, and disseminate these data sets in a meaningful fashion. With the consortium for functional glycomics (CFG) as the model system, this review discusses databases and the bioinformatics platform developed by this consortium to advance glycomics. PMID- 16478801 TI - Glycolipid activation of invariant T cell receptor+ NK T cells is sufficient to induce airway hyperreactivity independent of conventional CD4+ T cells. AB - Asthma is an inflammatory lung disease, in which conventional CD4+ T cells producing IL-4/IL-13 appear to play an obligatory pathogenic role. Here we show, in a mouse model of asthma, that activation of pulmonary IL-4/IL-13 producing invariant TCR+ CD1d-restricted natural killer T (NKT) cells is sufficient for the development of airway hyperreactivity (AHR), a cardinal feature of asthma, in the absence of conventional CD4+ T cells and adaptive immunity. Respiratory administration of glycolipid antigens that specifically activate NKT cells (alpha GalactosylCeramide and a Sphingomonas bacterial glycolipid) rapidly induced AHR and inflammation typically associated with protein allergen administration. Naive MHC class II-deficient mice, which lack conventional CD4+ T but have NKT cells, showed exaggerated baseline AHR and, when challenged with alpha GalactosylCeramide, demonstrated even greater AHR. These studies demonstrate an expanded role for NKT cells, in which NKT cells not only produce cytokines that influence adaptive immunity but also function as critical effector cells that can induce AHR. These results suggest that NKT cells responding to glycolipid antigens, as well as conventional CD4+ T cells responding to peptide antigens, may be synergistic in the induction of AHR, although in some cases, each may independently induce AHR. PMID- 16478802 TI - Optimization of ionic conductivity in doped ceria. AB - Oxides with the cubic fluorite structure, e.g., ceria (CeO2), are known to be good solid electrolytes when they are doped with cations of lower valence than the host cations. The high ionic conductivity of doped ceria makes it an attractive electrolyte for solid oxide fuel cells, whose prospects as an environmentally friendly power source are very promising. In these electrolytes, the current is carried by oxygen ions that are transported by oxygen vacancies, present to compensate for the lower charge of the dopant cations. Ionic conductivity in ceria is closely related to oxygen-vacancy formation and migration properties. A clear physical picture of the connection between the choice of a dopant and the improvement of ionic conductivity in ceria is still lacking. Here we present a quantum-mechanical first-principles study of the influence of different trivalent impurities on these properties. Our results reveal a remarkable correspondence between vacancy properties at the atomic level and the macroscopic ionic conductivity. The key parameters comprise migration barriers for bulk diffusion and vacancy-dopant interactions, represented by association (binding) energies of vacancy-dopant clusters. The interactions can be divided into repulsive elastic and attractive electronic parts. In the optimal electrolyte, these parts should balance. This finding offers a simple and clear way to narrow the search for superior dopants and combinations of dopants. The ideal dopant should have an effective atomic number between 61 (Pm) and 62 (Sm), and we elaborate that combinations of Nd/Sm and Pr/Gd show enhanced ionic conductivity, as compared with that for each element separately. PMID- 16478803 TI - On the origin and highly likely completeness of single-domain protein structures. AB - The size and origin of the protein fold universe is of fundamental and practical importance. Analyzing randomly generated, compact sticky homopolypeptide conformations constructed in generic simplified and all-atom protein models, all have similar folds in the library of solved structures, the Protein Data Bank, and conversely, all compact, single-domain protein structures in the Protein Data Bank have structural analogues in the compact model set. Thus, both sets are highly likely complete, with the protein fold universe arising from compact conformations of hydrogen-bonded, secondary structures. Because side chains are represented by their Cbeta atoms, these results also suggest that the observed protein folds are insensitive to the details of side-chain packing. Sequence specificity enters both in fine-tuning the structure and thermodynamically stabilizing a given fold with respect to the set of alternatives. Scanning the models against a three-dimensional active-site library, close geometric matches are frequently found. Thus, the presence of active-site-like geometries also seems to be a consequence of the packing of compact, secondary structural elements. These results have significant implications for the evolution of protein structure and function. PMID- 16478804 TI - Regulation of apoptosis by the p8/prothymosin alpha complex. AB - p8 is a small-stress protein involved in several cellular functions including apoptosis. To identify its putative partners, we screened a HeLa cDNA library by using the two-hybrid technique and found that p8 binds the antiapoptotic protein prothymosin alpha (ProTalpha). Fluorescence spectroscopy, circular dichroism, and NMR spectroscopy showed that p8 and ProTalpha formed a complex. Binding resulted in important changes in the secondary and tertiary structures of the proteins. Because p8 and ProTalpha form a complex, they could act in concert to regulate the apoptotic cascade. We induced apoptosis in HeLa cells by staurosporine treatment and monitored the effects of knocking down p8 and/or ProTalpha or overexpressing p8 and/or ProTalpha on caspase 3/7 and 9 activities and on cell death. Transfecting ProTalpha or p8 small interfering RNAs increased the activities of both caspases and the number of apoptotic nuclei. However, transfecting both small interfering RNAs resulted in no further increase. Overexpressing p8 or ProTalpha did not alter caspase activities, whereas overexpressing both resulted in a significant reduction of caspase activities. These results strongly suggest that the antiapoptotic response of HeLa cells upon staurosporine treatment requires expression of both p8 and ProTalpha. PMID- 16478806 TI - Paediatric utilization of an emergency department in Italy. AB - BACKGROUND: The purpose of this study was to determine the frequency and characteristics of paediatric attendance as a source of medically non-urgent problems at an accident and emergency department (ED) of a public non-teaching hospital in Crotone (Italy). METHODS: For each patient aged 16 years or younger, there were collected information on demographics and socioeconomic characteristics, medical history, route of referral, clinical complaints that they presented at the moment of their presentation at the ED, duration of presenting problems prior to arrival, hour of arrival, day of the week of arrival, and reason for attending the ED. Data about the consultation process and the final decision made were also recorded. RESULTS: Of a total of 980 patients included in the study, 27.6% had conditions that were definitely non-urgent. Multiple logistic regression analysis showed that the visit was non-urgent in younger population, in females, and in those attending the ED on the weekend. The results of the second multivariable regression analysis model indicate that patients who did not receive medical or surgical examination at the ED, with problems of longer duration prior to arrival at the ED, with non-traumatic injuries, and who did not require inpatient hospital admission were more likely to use the ED as a source of non-urgent care. The most frequent presenting problems for patient visits to ED were injury, respiratory diseases, and digestive symptoms. CONCLUSION: A closer cooperation within the health care organization system to provide a service responsive to the real needs of patients is essential. PMID- 16478807 TI - Mortality from external causes among ethnic German immigrants from former Soviet Union countries, in Germany. AB - BACKGROUND: Diaspora migration flows from the former Soviet Union to Western Europe and Israel have increased since the late 1980s. Risk factors responsible for the East-West mortality gap and post-migration factors may lead to higher mortality from external causes of death like suicide amongst such Diaspora immigrants. We investigated whether ethnic German immigrants from the former Soviet Union had increased mortality from external causes compared to native Germans. METHODS: We conducted a retrospective cohort study of 34 393 adults, so called Aussiedler who arrived in Germany's largest federal state between 1990 and 2001. We ascertained vital status and causes of death from registry data. Standardized mortality ratios (SMRs) were calculated using the native German population as comparison. Multivariate effects were assessed using Poisson regression. RESULTS: 1 657 members (4.8%) died, 88 from external causes. Overall SMR was 1.29 (95% confidence intervals 1.05-1.61). Males had a 39% higher mortality from all external causes and accidents, and a 30% higher mortality from suicide than German males. Females had slightly higher mortality from accidents but comparable mortality from all external causes. Aussiedler aged <65 years had rate ratios above one for external causes and accidents in multivariate models. SMRs for suicide and all external causes decreased with length of stay. CONCLUSION: While ethnic German immigrants have a mortality disadvantage compared to the NRW population, it is on a much lower scale than expected if they were representative of their source populations in former Soviet Union countries. PMID- 16478805 TI - Cytoprotective gene bi-1 is required for intrinsic protection from endoplasmic reticulum stress and ischemia-reperfusion injury. AB - Ischemia-reperfusion (IR) injury induces endoplasmic reticulum (ER) stress and cell death. Bax Inhibitor-1 (BI-1) is an evolutionarily conserved ER protein that suppresses cell death and that is abundantly expressed in both liver and kidney. We explored the role of BI-1 in protection from ER stress and IR injury by using bi-1 knockout mice, employing models of transient hepatic or renal artery occlusion. Compared to wild-type bi-1 mice, bi-1 knockout mice subjected to hepatic IR injury exhibited these characteristics: (i) increased histological injury; (ii) increased serum transaminases, indicative of more hepatocyte death; (iii) increased percentages of TUNEL-positive hepatocytes; (iv) greater elevations in caspase activity; and (v) more activation of ER stress proteins inositol-requiring enzyme 1 and activating transcription factor 6 and greater increases in expression of ER stress proteins C/EBP homologous protein and spliced XBP-1 protein. Moreover, hepatic IR injury induced elevations in bi-1 mRNA in wild-type liver, suggesting a need for bi-1 gene induction to limit tissue injury. Similar sensitization of kidney to ER stress and IR injury was observed in bi-1(-/-) mice. We conclude that bi-1 provides endogenous protection of liver and kidney from ER stress and IR injury. Analysis of components of the bi-1-dependent pathway for protection from IR injury may therefore reveal new strategies for organ preservation. PMID- 16478808 TI - Frequency of serum creatinine monitoring during allopurinol therapy in ambulatory patients. AB - BACKGROUND: Allopurinol dosage reduction is recommended in patients with renal dysfunction because drug toxicity risk is increased. Little information is available about serum creatinine (SCr) monitoring in ambulatory patients taking allopurinol. OBJECTIVE: To evaluate SCr monitoring among patients prescribed allopurinol, identify associated factors, and evaluate administrative data in assessing monitoring. METHODS: Information for this retrospective cohort study was drawn from a dataset of 2 020 037 individuals; approximately 200 000 members from each of 10 organizations. Study patients had received at least one year of ongoing allopurinol prescription dispensings. Patient variables analyzed included age, gender, chronic diseases, outpatient visits, hospitalizations, gout diagnosis, and SCr monitoring. A random sample of medical records was reviewed to assess the accuracy of the automated data. Statistical analysis included descriptive and logistic regression techniques. RESULTS: Overall, 1139 (26%) of 4357 patients did not have SCr monitoring. For individuals without recent hospitalization, factors protective against lack of monitoring were increasing age (OR 0.77 per 10 y; 95% CI 0.74 to 0.79), more chronic diseases (OR 0.81; 95% CI 0.78 to 0.83), more outpatient visits (OR 0.87 per 5 visits; 95% CI 0.83 to 0.91), and gout diagnosis (OR 0.74; 95% CI 0.65 to 0.85). The sensitivity and specificity of administrative data compared with medical records for SCr monitoring were 92% and 65%, respectively. CONCLUSIONS: More than one-fourth of patients dispensed allopurinol did not have SCr monitoring during one year of therapy. Lack of monitoring and lack of subsequent possible dosage adjustment put patients at increased risk of allopurinol toxicity. PMID- 16478810 TI - The development of pharmacodynamics as a pharmaceutical science: a personal perspective. PMID- 16478809 TI - Tolerance of vancomycin for surgical prophylaxis in patients undergoing cardiac surgery and incidence of vancomycin-resistant enterococcus colonization. AB - BACKGROUND: In 2001, vancomycin replaced cefuroxime for antibiotic prophylaxis in patients undergoing cardiac surgery at our institution due to high rates of surgical site infections caused by methicillin-resistant Staphylococcus spp. However, few data supported the use of vancomycin for surgical prophylaxis. OBJECTIVE: To determine the tolerance of vancomycin for antibiotic prophylaxis and incidence of vancomycin-resistant Enterococcus (VRE) in cardiac surgery patients. METHODS: In 2 separate studies, we assessed the adverse effects in patients given perioperative vancomycin (study 1) and the incidence of VRE in patients given perioperative vancomycin (study 2). Study 1 was a prospective cohort study of patients undergoing coronary artery bypass graft (CABG) or valve replacement surgery given vancomycin (1 dose preoperatively/2 doses postoperatively) for antibiotic prophylaxis between October 2003 and December 2004. Patients were assessed for tolerance to the antibiotic regimen. In study 2, cardiac surgery patients receiving perioperative vancomycin were screened for VRE before therapy and at day 7 of hospitalization. VRE was detected using standard microbiologic procedures. RESULTS: In study 1, 1161 patients (CABG = 75%; valve = 19%; both = 6%) were evaluated. All patients but one (99.9%) were prescribed preoperative vancomycin. Therapy was changed for 34 (2.9%) patients, of which 20 changes were due to physician preference for another antibiotic. The only toxicity that required a change in the vancomycin regimen was red man's syndrome, which was experienced by 9 (0.8%) patients. Four patients did not receive a second postoperative dose due to prior renal insufficiency. Patients were most commonly switched to cefuroxime (n = 26), linezolid (n = 2), cefepime (n = 2), gatifloxacin, cefazolin, levofloxacin, or ceftriaxone (n = 1, each). In study 2, 100 patients were screened for the emergence of VRE colonization. No patient was VRE positive at baseline and 4 (4%) were positive at day 7. CONCLUSIONS: Surgical antibiotic prophylaxis with vancomycin was reasonably well tolerated in CABG and valve replacement surgery, with a 4% incidence of VRE colonization. PMID- 16478811 TI - Genetic predisposition to neuroleptic malignant syndrome in siblings. PMID- 16478812 TI - Quetiapine for insomnia associated with refractory depression exacerbated by phenelzine. AB - OBJECTIVE: To report the successful treatment of phenelzine-associated insomnia with low-dose quetiapine in a patient with refractory depression. CASE SUMMARY: A 42-year-old white man with severe major depression unresponsive to selective serotonin-reuptake inhibitors, bupropion, and tricyclic antidepressants improved following treatment with the monoamine oxidase inhibitor (MAOI) phenelzine. Insomnia, present to a moderate degree prior to antidepressant therapy, worsened markedly following phenelzine treatment and failed to respond to diphenhydramine, temazepam, triazolam, clonazepam, zolpidem, or trazodone given at high therapeutic doses. Sleep disturbance resolved with low-dose (50 mg) adjunctive quetiapine, with no adverse effects. DISCUSSION: Major depression refractory to standard therapy is a common and serious condition. Some cases respond to MAOIs; however, orthostatic hypotension and insomnia frequently occur. Potentially serious MAOI interactions with psychotropic drugs have raised concerns about combining these agents. In this case, a failure of a number of other medications known to treat MAOI-associated insomnia safely prompted a trial of quetiapine. Despite the possibility that enhanced serotonergic activity might have resulted in serotonin syndrome, no adverse interactions between phenelzine and quetiapine were noted. The use of low-dose, once-daily quetiapine, along with its unique binding properties, may account for its increased safety in combination with phenelzine. CONCLUSIONS: This case illustrates that low-dose quetiapine may be an alternative treatment for phenelzine-associated insomnia. Further case reports are needed to establish the safety and effectiveness of combining these agents. PMID- 16478813 TI - Pharmacoeconomics: historical perspective. PMID- 16478814 TI - Effects of testosterone replacement in androgen-deficient women with hypopituitarism: a randomized, double-blind, placebo-controlled study. AB - CONTEXT: Hypopituitarism in women is characterized by profound androgen deficiency due to a loss of adrenal and/or ovarian function. The effects of testosterone replacement in this population have not been reported. OBJECTIVE: The objective of the study was to determine whether physiologic testosterone replacement improves bone density, body composition, and/or neurobehavioral function in women with severe androgen deficiency secondary to hypopituitarism. DESIGN: This was a 12-month randomized, placebo-controlled study. SETTING: The study was conducted at a general clinical research center. STUDY PARTICIPANTS: Fifty-one women of reproductive age with androgen deficiency due to hypopituitarism participated. INTERVENTION: Physiologic testosterone administration using a patch that delivers 300 microg daily or placebo was administered. MAIN OUTCOME MEASURES: Bone density, fat-free mass, and fat mass were measured by dual x-ray absorptiometry. Thigh muscle and abdominal cross sectional area were measured by computed tomography scan. Mood, sexual function, quality of life, and cognitive function were assessed using self-administered questionnaires. RESULTS: Mean free testosterone increased into the normal range during testosterone administration. Mean hip (P = 0.023) and radius (P = 0.007), but not posteroanterior spine, bone mineral density increased in the group receiving testosterone, compared with placebo, as did mean fat-free mass (P = 0.040) and thigh muscle area (P = 0.038), but there was no change in fat mass. Mood (P = 0.029) and sexual function (P = 0.044) improved, as did some aspects of quality of life, but not cognitive function. Testosterone at physiologic replacement levels was well tolerated, with few side effects. CONCLUSIONS: This is the first randomized, double-blind, placebo-controlled study to show a positive effect of testosterone on bone density, body composition, and neurobehavioral function in women with severe androgen deficiency due to hypopituitarism. PMID- 16478815 TI - Clinical review: The rationale for banning human chorionic gonadotropin and estrogen blockers in sport. AB - CONTEXT: The objective of the study was to review the rationale underlying the banning of human chorionic gonadotropin (hCG) and estrogen blockers (antiestrogens, specific estrogen receptor modulators, aromatase inhibitors) in sports for male and female athletes in the light of gender differences in regulation of reproductive physiology. EVIDENCE ACQUISITION: We reviewed well controlled clinical studies of exogenous testosterone effects on human muscle size and strength in men and all available evidence relevant to the effects of hCG and estrogen blockers on blood testosterone in men and women. EVIDENCE SYNTHESIS: Well-designed placebo-controlled clinical studies in men with suppressed pituitary-testicular axis establish a strong case that, across a wide range from sub- to supraphysiological doses, muscle growth and strength is proportional to exogenous testosterone dose and resulting blood testosterone concentrations. In men, there is unequivocal evidence that hCG and estrogen blockers cause consistent and sustained rise in blood testosterone concentrations. In women, although there has been no direct testing of ergogenic or myotrophic properties of exogenous testosterone in healthy women, either hCG or estrogen blockers do not produce any consistent or biologically significant increase blood testosterone concentrations. CONCLUSIONS: In men undergoing potential stimulation of endogenous blood testosterone concentrations, blood testosterone concentration is a reasonable surrogate measure for muscle growth and increased strength in men. Because hCG and estrogen blockers produce marked increase in blood testosterone concentration in men, this provides strong evidence to support the banning of hCG and estrogen blockers in men. In women, however, the negligible effect on blood testosterone suggests that drug-induced performance enhancement by hCG or estrogen blockers is highly unlikely. Furthermore, routine urinary hCG testing in young women risks invasion of privacy by detecting unrecognized pregnancy. These considerations suggest that prohibition of hCG and estrogen blockers should be restricted to men in which they are well justified. PMID- 16478816 TI - Disordered adrenocorticotropin secretion in women with major depression. AB - CONTEXT: Major depression is accompanied by activation of the hypothalamic pituitary axis and evidence of abnormalities in circadian and ultradian hormone rhythms. In addition, diminished negative feedback of cortisol on ACTH has been found. OBJECTIVE: The objective of the study was to compare ACTH and cortisol hormonal patterns in women with major depression with normal control women. DESIGN: This was a case control study. SETTING: The study was conducted at a general clinical research center. PATIENTS AND OTHER PARTICIPANTS: Healthy, drug free, premenopausal women with major depression and age and menstrual cycle day matched healthy control women were included in the study. MAIN OUTCOME MEASURE: ACTH and cortisol data were measured every 10 min for 24 h analyzed with approximate entropy and cross-approximate entropy to determine orderliness of hormone secretion and relationship between ACTH and cortisol in terms of feedforward and feedback synchrony. RESULTS: Depressed women manifested increased approximate entropy, indicating more disorderly secretion, of ACTH and elevated forward cross-approximate entropy of ACTH on cortisol, denoting unopposed ACTH drive. CONCLUSIONS: These data support other evidence of hormonal rhythm abnormalities in depression and are compatible with accentuated feedforward drive by ACTH. PMID- 16478817 TI - Clinical and histological characteristics of papillary thyroid microcarcinoma: results of a retrospective study in 243 patients. AB - CONTEXT: The recognition of thyroid microcarcinoma has increased due to the widespread use of ultrasound-guided fine-needle aspiration biopsies. OBJECTIVE: The objective of this study was to describe histological and clinical characteristics of papillary thyroid microcarcinoma (PTMC) less than or equal to 1 cm. DESIGN: This study was a retrospective cohort. SETTING: This study was conducted at a university hospital endocrine clinic. PATIENTS: Over a 9-yr period, 243 consecutive patients with PTMC were studied. RESULTS: PTMC was an incidental finding at surgery in 21.4% of the PTMC cases. There were no differences in the clinical characteristics between those with incidental PTMC and those with suspected thyroid carcinoma. None of the patients with a cancer less than 8 mm had distant metastases, whereas distant metastases were observed in patients with cancers >/= 8 mm (P or = 8 mm Hg, vasopressors targeting mean arterial pressure > or = 65 mm Hg, and (if necessary) packed RBCs (PRBCs) and/or dobutamine targeting central venous oxygen saturation > or = 70%. A retrospective analysis was performed of emergency department (ED) patients with persistent sepsis-induced hypotension (systolic BP < 90 mm Hg despite 1.5 L of IVF) treated with EGDT during the first year of the initiative. Primary outcome measures included successful achievement of EGDT end points and time to achievement. A secondary analysis was performed comparing EGDT cases to historical control cases (nonprotocolized control subjects without invasive monitoring). RESULTS: All end points were achieved in 20 of 22 cases (91%). The median time to reach each end point was < or = 6 h. In the secondary analysis, patients (n = 38; EGDT, n = 22; pre-EGDT, n = 16) had similar age, do-not-resuscitate status, severity scores, hypotension duration, and vasopressor requirement (p = not significant). In the ED, EGDT used more IVF and included PRBC/dobutamine utilization, without any impact on the overall use of these therapies through the first 24 h in the ICU. EGDT was associated with decreased ICU pulmonary artery catheter (PAC) utilization (9.1% vs 43.7%, p = 0.01). CONCLUSIONS: With effective emergency medicine/critical care collaboration, we demonstrate that EGDT end points can reliably be achieved in real-world sepsis resuscitation. ED-based EGDT appears to decrease ICU PAC utilization. PMID- 16478836 TI - Serum cardiovascular risk factors in obstructive sleep apnea. AB - BACKGROUND: Obstructive sleep apnea (OSA) patients have increased cardiovascular morbidity and mortality. The cardiovascular markers associated with OSA are currently not defined. OBJECTIVES: The aims of this study were to determine whether OSA is associated with serum cardiac risk markers and to investigate the relationship between them. METHODS: Sixty-two male patients were classified into two groups with respect to apnea-hypopnea index (AHI): group 1, sleep apnea (n = 30), with AHI > 5; and group 2 (n = 32), with AHI < 5. We compared cardiovascular risk factors in both groups with control subjects (n = 30) without OSA (AHI < 1). Serum cholesterol, triglyceride, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), apolipoprotein A-I, apolipoprotein B, lipoprotein (a), C-reactive protein (CRP), and homocysteine were measured. Statistical significance was assessed with analysis of variance at p < 0.05. In correlation analysis, Pearson correlation was used. RESULTS: There was no significant difference between group 1 and group 2 in total cholesterol, LDL-C, HDL-C, triglyceride, apolipoprotein A-I, apolipoprotein B, and lipoprotein (a). All of the M-mode echocardiographic parameters were in the normal reference range. Serum homocysteine and CRP levels were significantly increased in group 1 compared to group 2 (p < 0.05). Serum CRP values were increased in both group 1 and group 2 when compared with control subjects (p < 0.05). Serum homocysteine values were higher in group 1 than in control subjects (p < 0.05). CONCLUSIONS: Our results show that OSA syndrome is associated not only with slight hyperhomocysteinemia but also with increased CRP concentrations. Increased plasma concentrations of homocysteine and CRP can be useful in clinical practice to be predictor of long-term prognosis for cardiovascular disease and the treatment of OSA. PMID- 16478837 TI - Improvement in nocturnal disordered breathing after first-ever ischemic stroke: role of dysphagia. AB - STUDY OBJECTIVE: The aim of this study was to analyze the role of dysphagia as a model of pharyngeal muscle dysfunction in the time course of nocturnal disordered breathing (NDB) in patients who experienced a first-ever ischemic stroke. DESIGN: Prospective study. PATIENTS AND INTERVENTIONS: Fifty-nine consecutive patients (mean age, 73.2 years; SD, 12.8 years) were studied. Clinical sleep and neurologic data and vascular risk factors were recorded. Two nocturnal studies using a portable autotitration device (AutoSet Portable Plus II system; ResMed; Sydney, NSW, Australia) were performed in both the acute phase (mean duration, 1.23 days; SD, 0.7 day) and the stable phase (mean duration 65.9 days; SD, 12.5 days) of the neurologic event in all patients. RESULTS: The mean total apnea hypopnea index (AHI) measured with the autotitration device in the acute phase was 34.9 (SD, 25.2) vs 20.1 (SD, 21.7) in the stable phase, both with predominance of obstructive apnea. Patients with dysphagia (n = 30) showed the largest number of obstructive apneic episodes (OAIs) in the acute phase (AHI, 40 episodes; OAI, 30.4 episodes), with a significant reduction in this type of apnea during the stable phase of stroke (AHI, 24.7 episodes; OAI, 17.7 episodes), coinciding with the recovery of pharyngeal muscle function. In contrast, nondysphagic patients (n = 29) showed no significant changes in NDB from the acute to the stable phase of stroke. Logistic regression analysis found dysphagia to be the best independent predictor of AHI reduction of > 50% from baseline (odds ratio, 13.4; 95% confidence interval, 3.3 to 39.6; p = 0.001). CONCLUSION: The present study shows significant improvement in the number obstructive apneic events occurring in the stable phase of a first-ever ischemic stroke in patients with transient pharyngeal muscle alterations secondary to the neurologic lesion. PMID- 16478838 TI - Budesonide/formoterol in a single inhaler for maintenance and relief in mild-to moderate asthma: a randomized, double-blind trial. AB - STUDY OBJECTIVE: To compare a novel asthma management strategy- budesonide/formoterol in a single inhaler for both maintenance therapy and symptom relief--with a higher dose of budesonide plus as-needed terbutaline. METHODS: This was a 6-month, randomized, double-blind, parallel-group study in patients with mild-to-moderate asthma (n = 697; mean age, 38 years [range, 11 to 79 years]; mean baseline FEV1, 75% of predicted; mean inhaled corticosteroid [ICS] dosage, 348 microg/d). Following a 2-week run-in period, all patients received two blinded, dry powder inhalers, one containing maintenance medication and one containing medication to be used as needed for the relief of symptoms. Patients were randomized to receive either budesonide/formoterol (80 microg/4.5 microg, two inhalations qd) for maintenance plus additional inhalations as needed for symptom relief, or budesonide (160 microg, two inhalations qd) for maintenance medication plus terbutaline (0.4 mg) as needed. The primary efficacy variable was morning peak expiratory flow (PEF). RESULTS: Patients receiving budesonide/formoterol showed greater improvements in morning PEF than patients receiving budesonide (increases of 34.5 L/min vs 9.5 L/min, respectively; p < 0.001). The risk of having a severe exacerbation (hospitalization/emergency department [ED] treatment, oral steroids for asthma, or a > or = 30% decrease from baseline in morning PEF on 2 consecutive days) was 54% lower with budesonide/formoterol vs budesonide (p = 0.0011). Budesonide/formoterol patients experienced 90% fewer hospitalizations/ED treatments due to asthma than budesonide patients (1 vs 10, respectively; p = 0.026). The increased efficacy with budesonide/formoterol was achieved with less ICS than was used in the budesonide group (mean dose, 240 microg/d vs 320 microg/d, respectively) and with 77% fewer oral steroid treatment days vs budesonide (114 days vs 498 days, respectively). Both treatments were well tolerated. CONCLUSIONS: Budesonide/formoterol for both maintenance and relief improves asthma control with a lower steroid load compared with a higher dose of budesonide plus terbutaline. PMID- 16478840 TI - A comparative study of two angiogenic factors: vascular endothelial growth factor and angiogenin in induced sputum from asthmatic children in acute attack. AB - BACKGROUND: Angiogenesis is a prerequisite for airway remodeling in bronchial asthma. Several factors may play important roles in inflammation and angiogenesis through effects on inflammatory cell infiltration or neovascularization. OBJECTIVES: (1) To determine the levels of vascular endothelial growth factor (VEGF) and angiogenin in sputum supernatants of asthmatic children during the acute attack and 6 weeks after start of therapy; and (2) to correlate their levels with the degree of asthma severity. SUBJECTS AND METHODS: Twenty asthmatic children with acute attack (mean age, 9.6 +/- 3.5 years [+/- SD]) and 12 sex- and age-matched healthy control children were enrolled in the study. Sputum supernatants were collected for determination of VEGF and angiogenin levels. Serum samples were withdrawn for IgE measurement. The above tests were performed using an enzyme-linked immunosorbent assay. The FEV1 was measured using spirometry. VEGF, angiogenin, and FEV1 estimations were repeated for asthmatic children 6 weeks after start of therapy. RESULTS: During the acute attack, asthmatic children had significantly higher levels of VEGF and angiogenin than in healthy control children (p < 0.001). VEGF and angiogenin levels showed more elevation with increase in asthma severity (p < 0.001). A significant positive correlation existed between both angiogenic factors (r = 0.98, p < 0.001). A negative significant correlation was found between FEV1 percentage of predicted and both VEGF (r = -0.99, p < 0.001) and angiogenin (r = -0.97, p < 0.001). A nonsignificant correlation was found between serum IgE and sputum VEGF (r = 0.09, p > 0.05). Although there was a significant decrease in the levels of both VEGF and angiogenin after 6 weeks of treatment with corticosteroid inhalation therapy, the levels did not reach normal control levels (p < 0.001 and p < 0.05, respectively). CONCLUSIONS: Our results show that both VEGF and angiogenin levels were elevated in children with acute asthma. The study also suggests that increased severity of bronchial asthma in children is associated with the expression of both angiogenic factors, which are implicated in asthma pathogenesis. After 6 weeks of therapy, the levels of both angiogenic factors showed significant decrease. PMID- 16478839 TI - Randomized controlled trial of emergency department interventions to improve primary care follow-up for patients with acute asthma. AB - OBJECTIVE: Emergency department (ED) visits for asthma are frequent and may indicate increased morbidity and poor primary care access. Our objective was to compare the effect of two interventions on primary care follow-up after ED treatment for asthma exacerbations. METHODS: We performed a randomized controlled trial of patients 2 to 54 years old who were judged safe for discharge receiving prednisone, and who were available for contact at 2 days and 30 days. Patients were excluded if they were previously enrolled or did not speak English. Patients received usual discharge care (group A); free prednisone, vouchers for transport to and from a primary care visit, and either a telephone reminder to schedule a visit (group B); or a prior scheduled appointment (group C). Follow-up with a primary care provider for asthma within 30 days was the main outcome. Secondary outcomes were recurrent ED visits, subsequent hospitalizations, quality of life, and use of inhaled corticosteroids 1 year later. RESULTS: Three hundred eighty four patients were enrolled. Baseline demographics, chronic asthma severity, and access to care were similar across groups. Primary care follow-up was higher in group C (65%) vs group A (42%) or group B (48%) [p = 0.002]. Group C intervention remained significant (odds ratio, 2.8; 95% confidence interval, 1.5 to 5.1) when adjusted for other factors influencing follow-up (prior primary care relationship, insurance status). There were no differences in ED, hospitalizations, quality of life, or inhaled corticosteroid use at 1 year after the index ED visit. CONCLUSION: An intervention including free medication, transportation vouchers, and appointment assistance significantly increased the likelihood that discharged asthma patients obtained primary care follow-up but did not impact long-term outcomes. PMID- 16478841 TI - Percentage fall in FVC at the provocative concentration of methacholine causing a 20% fall in FEV1 in symptomatic asthma and clinical remission during adolescence. AB - BACKGROUND: Many children with asthma go into long-term clinical remission at adolescence, but bronchial hyperresponsiveness (BHR) persists in approximately one half of these subjects. BHR is usually assessed by measuring the provocative concentration of methacholine causing a 20% fall in FEV1 (PC20). The percentage fall in FVC at the PC20 (deltaFVC) has been suggested to be a more useful index of disease severity in asthma than PC20. STUDY OBJECTIVE: The aim of this study was to determine whether deltaFVC is higher in adolescents with symptomatic asthma than in those with clinical remission. PATIENTS AND METHODS: Forty adolescents with symptomatic asthma and 80 adolescents with asthma remission underwent methacholine challenge testing. DeltaFVC and PC20 were measured on the methacholine dose-response curve. RESULTS: The mean (95% confidence interval [CI]) deltaFVC (15.5% [95% CI, 14.1 to 16.9%]) in the symptomatic group (n = 40) was significantly higher (p = 0.017) than that (12.8% [95% CI, 11.5 to 14.1%]) in the BHR-positive (PC20 < 16 mg/mL) remission group (n = 44) or that (11.5% [95% CI, 10.2 to 12.8%]) of the BHR-negative remission group (n = 36), with no difference between the two latter groups (p = 0.581). No significant correlation was found between deltaFVC and PC20 in the symptomatic group (r = -0.156, p = 0.336) or in the whole remission group (r = -0.187, p = 0.097). CONCLUSIONS: Adolescents with symptomatic asthma had a higher deltaFVC than those with clinical remission, irrespective of the presence of BHR in the latter group. This finding suggests that deltaFVC may serve as an adjunct marker for differentiating between asthma persistence and remission during adolescence. PMID- 16478842 TI - Heart rate variability biofeedback: effects of age on heart rate variability, baroreflex gain, and asthma. AB - OBJECTIVES: To present additional analysis of data from a previously published study showing that biofeedback training to increase heart rate variability (HRV) can be an effective component in asthma treatment. HRV and intervention-related changes in HRV are negatively correlated with age. Here we assess the effects of age on biofeedback effects for asthma. DESIGN: Ten sessions of HRV biofeedback were administered to 45 adults with asthma. Medication was prescribed by blinded physicians according to National Heart, Lung, and Blood Institute criteria. Medication needs were reassessed biweekly. RESULTS: Decreases in need for controller medication were independent of age. There were larger acute decreases in forced oscillation frequency dependence in the older group but larger increases in HRV variables in the younger group. Differences between age groups were smaller among subjects trained in pursed-lips abdominal breathing as well as biofeedback, than among those receiving only biofeedback. CONCLUSIONS: Age related attenuation of biofeedback effects on cardiovascular variability does not diminish the usefulness of the method for treating asthma among older patients. Additional training in pursed-lips abdominal breathing obliterates the effects of age on HRV changes during biofeedback. PMID- 16478843 TI - Coexistent chronic conditions and asthma quality of life: a population-based study. AB - OBJECTIVE: Reports of the prevalence and impact of comorbid conditions among people with asthma have been limited to certain population groups or convenience samples. Our aim was to examine the prevalence of major comorbidity in asthma and associations with quality of life and functional status in the general population. STUDY DESIGN/SETTING: The WANTS Health and Well-being Survey is a cross-sectional representative population household telephone interview survey in three Australian states. PARTICIPANTS: Representative sample of noninstitutionalized adults in three Australian states. MEASUREMENT AND RESULTS: From the available sample of 10,080 patients, 7,619 interviews were completed (participation rate, 74.8%), with 834 people reporting current doctor-diagnosed asthma (11.2%). People with asthma were more likely to report one of the selected comorbid conditions: diabetes, arthritis, heart disease, stroke, cancer, osteoporosis (adjusted odds ratio, 1.9; 95% confidence interval, 1.5 to 2.2). Among people with asthma, there were statistically and clinically significant decreases in usual activity levels and in Short Form-12 physical component summary scores when another chronic condition was also present. For those with any of the chronic conditions, the additional presence of asthma was associated with significant further impairment in quality of life in those aged > 35 years but not in younger adults. CONCLUSION: The significant reduction in quality of life associated with comorbidity in asthma has implications for disease management and organization of care, as well as for the design and external validity of single-disease clinical trials. PMID- 16478844 TI - An interdisciplinary intervention for undertreated pediatric asthma. AB - STUDY OBJECTIVES: To examine the effectiveness of an interdisciplinary intervention for pediatric asthma. DESIGN: Randomized, controlled study. SETTING: Urban tertiary-referral pediatric hospital. PARTICIPANTS: One hundred seventy five patients with asthma lacking written treatment plans and presenting with asthma-related emergency department visits (two or more) and/or hospitalizations (one or more) in the past year were randomized to a comparison group receiving medical care alone (n = 86) or to an interdisciplinary intervention group receiving medical care, asthma education, and problem-solving therapy (n = 89) INTERVENTION: All participants received written asthma management plans, peak flow meters, and spacer devices. The intervention group also received asthma education, an asthma risk profile assessment, brief problem-solving therapy, and access to a 24-h nurse advice line. The primary outcome measure was change in asthma symptoms, and secondary outcomes included health-care utilization and asthma-related quality of life. RESULTS: Both groups demonstrated significant reductions in asthma symptoms and improvements in quality of life without any between-group differences identified over the course of follow-up. In contrast, the intervention group demonstrated less frequent health-care utilization than the comparison group, with 28% of the intervention group requiring emergency department or inpatient services for asthma compared to 41% of the comparison group (adjusted odds ratio, 1.92; 95% confidence interval, 1.00 to 3.69) over the 12-month follow-up period. CONCLUSIONS: This study examined the effectiveness of an interdisciplinary intervention for undertreated asthma. The intervention did not result in improvements in asthma symptoms, but accomplished modest reductions in the utilization of acute medical care. PMID- 16478845 TI - Home exposures, parental atopy, and occurrence of asthma symptoms in adulthood in southern Taiwan. AB - OBJECTIVE: Parental atopy and environmental exposures at home have been recognized risk factors for adulthood asthma. However, the relative contributions of specific risk factors and the overall contributions of heredity or home exposure remain unexplored. The purpose of this study was to identify predictors and estimate the population attributable risk (PAR) of each exposure for typical asthma symptoms among 26- to 50-year-old Taiwanese. We also investigated whether an interactive effect existed between parental atopy and home exposures on the occurrence of asthma symptoms in adulthood. DESIGN: A cross-sectional study with retrospective components. SETTING: Elementary and middle schools in Southern Taiwan. SUBJECTS: Between March and October 2004, we conducted a cross-sectional survey among schoolchildren's parents from 94 elementary and middle schools in Southern Taiwan. The main outcome measure was typical asthma-like symptoms occurring within the preceding 5 years. Information on hereditary and home exposures was collected by using a self-administered questionnaire. RESULTS: After excluding unqualified questionnaires, data from 24,784 subjects were left for analysis. New-onset asthma was reported for 0.83% of male (n = 80 of 9,662) and 1.36% of female subjects (n = 206 of 15,122). Besides parental atopic factors, environmental tobacco smoke or pet avoidance and visible mold on walls at home were independently associated with the occurrence of asthma symptoms. Mutually adjusted models produced statistically significant associations between any home exposure (odds ratio [OR], 1.80; 95% confidence interval [CI], 1.08 to 3.23; PAR, 28.04%), parental atopy (OR, 4.47; 95% CI, 3.47 to 5.75; PAR, 31.38%), and new-onset asthma. However, there was no interaction between parental atopy and home exposures. CONCLUSIONS: Home exposures and parental atopy both increased the risks of new-onset asthma in adulthood but did not show an interactive effect. These two exposure categories approximately contributed equally to the adulthood asthma. PMID- 16478846 TI - Risk factors for onset of asthma: a 12-year prospective follow-up study. AB - BACKGROUND: Asthma development and prognosis have been studied extensively in at risk populations, but knowledge of the natural history of asthma in the general population is limited. OBJECTIVE: To describe the incidence and remission of asthma and its predictors, data from a 12-year follow-up study of a random population sample (n = 291; age range, 7 to 17 years) at enrollment, were analyzed. METHODS: The sample was examined in 1986 and in 1998. A case history, including data on asthma, allergic diseases, and lifestyle factors, was obtained by questionnaire and interview. Airway hyperresponsiveness (AHR) to histamine, lung function, and skin-prick test reactivity to a standard panel of 10 aeroallergens were measured. RESULTS: The point prevalence of asthma increased from 4.1% at the first survey to 11.7% at follow-up, at which point 19.6% of the sample had ever experienced asthma symptoms. Of the subjects with ever-asthma, 40% had remitted at follow-up. Asthma developed in 45 subjects (16.1%) during the follow-up period, which was predicted by the following factors: wheezing in childhood (odds ratio [OR], 3.61; 95% confidence interval [CI], 1.34 to 9.75), AHR (OR, 4.94; 95% CI, 2.42 to 10.08), allergic sensitization to house dust mites (OR, 3.23; 95% CI, 1.00 to 10.40), and dermatitis (OR, 2.94; 95% CI, 1.22 to 7.11). The simultaneous presence of more than one of these risk factors was associated with a high probability of developing asthma at follow-up (61.5%). In subjects without any risk factors, such as AHR, allergic sensitization, rhinitis, dermatitis or wheezing in childhood; paternal/maternal allergy, or asthma, asthma developed in only 4% during follow-up. CONCLUSION: The presence of AHR and concomitant atopic manifestations in childhood increase the risk of developing asthma in adulthood, and should be recognized as markers of prognostic significance, whereas the absence of these manifestations predicts a very low risk of future asthma. PMID- 16478847 TI - Effect of interactions between lower airway bacterial and rhinoviral infection in exacerbations of COPD. AB - STUDY OBJECTIVES: The inflammatory responses and associated clinical severity of COPD exacerbations are greatly variable, and the determinants of these factors are poorly understood. We examined the hypothesis that bacteria and viruses may modulate this heterogeneity and that interactions between bacterial and viral infection may affect changes in airway bacterial load and the clinical features and inflammatory responses of exacerbations in patients with COPD. DESIGN: Prospective cohort study. SETTING: Outpatient Department, London Chest Hospital, London, UK. PATIENTS: Thirty-nine patients with COPD. MEASUREMENTS: We prospectively studied 56 COPD exacerbations, obtaining clinical data and paired sputum and serum samples at baseline and exacerbation. Qualitative and quantitative microbiology, polymerase chain reaction detection for rhinovirus, and estimation of cytokine levels by enzyme-linked immunosorbent assay were performed. RESULTS: A total of 69.6% of exacerbations were associated with a bacterial pathogen, most commonly Haemophilus influenzae. Rhinovirus was identified in 19.6% of exacerbations. The rise in bacterial load at exacerbation correlated with the rise in sputum interleukin (IL)-8 (r = 0.37, p = 0.022) and fall in FEV1 (r = 0.35, p = 0.048). Exacerbations with both rhinovirus and H. influenzae had higher bacterial loads (10(8.56) cfu/mL vs 10(8.05)cfu/mL, p = 0.018) and serum IL-6 (13.75 pg/mL vs 6.29 pg/mL, p = 0.028) than exacerbations without both pathogens. In exacerbations with both cold symptoms (a marker of putative viral infection) and a bacterial pathogen, the FEV1 fall was greater (20.3% vs 3.6%, p = 0.026) and symptom count was higher (p = 0.019) than those with a bacterial pathogen alone. CONCLUSIONS: The clinical severity and inflammatory responses in COPD exacerbations are modulated by the nature of the infecting organism: bacterial and viral pathogens interact to cause additional rises in inflammatory markers and greater exacerbation severity. PMID- 16478848 TI - Response to one-legged cycling in patients with COPD. AB - BACKGROUND: In patients with COPD, exercise intensity is often limited by the ventilatory system. We hypothesized that by exercising with a smaller muscle mass, ventilatory-limited patients would perform more high-intensity, muscle specific work. The study objectives were as follows: (1) to determine the limitations of exercising with a smaller muscle mass, compared with conventional two-legged exercise; and (2) to determine the endurance time, using the same muscle-specific intensity, during one-legged vs two-legged exercise. METHODS: Nine patients (mean +/- SD FEV1, 36 +/- 13% of predicted) completed incremental exercise, and nine other patients (mean FEV1, 42 +/- 16% of predicted) completed constant-power exercise. Nine healthy subjects (FEV1, 104 +/- 14% of predicted) completed both tests. All subjects completed tests using two-legged and one legged pedaling. RESULTS: Peak oxygen uptake (VO2peak) was similar during one legged and two-legged incremental exercise among patients (difference, 0.03 L/min; 95% confidence interval [CI], -0.10 to 0.16 L/min; p = 0.60), as were ventilation and dyspnea scores. VO2peak was lower during one-legged vs two-legged exercise (-0.57 mL/min; 95% CI, -0.81 to -0.32 mL/min; p < 0.001) among healthy subjects with substantial ventilatory and heart rate reserve. Patients endured one-legged pedaling at a constant power longer than two-legged pedaling (16.97 min; 95% CI, 9.98 to 23.96 min; p < 0.001), resulting in greater work (12.48 kilojoules [kJ]; 95% CI, 2.58 to 22.39 kJ; p = 0.02). Healthy subjects completed similar work (-4.02 kJ; 95% CI, -18.59 to 10.55 kJ; p = 0.54) with one-legged vs two-legged pedaling. CONCLUSION: These observations demonstrate the effectiveness of using one-legged exercise at the same muscle-specific intensity in extending the duration of exercise among patients with COPD. This has important implications for training approaches designed to enhance exercise function among ventilatory-limited patients. PMID- 16478849 TI - Pulmonary cryptococcosis: comparison of clinical and radiographic characteristics in immunocompetent and immunocompromised patients. AB - STUDY OBJECTIVES: We compared the clinical characteristics and imaging findings between immunocompetent and immunocompromised patients in whom pulmonary cryptococcosis had been diagnosed to define the role of serum cryptococcal antigen (sCRAG) and radiographs during a follow-up period of up to 1 year. DESIGN: Retrospective cohort study. SETTING: University hospital. PATIENTS: The clinical records, chest radiographs, and CT scan findings of 13 immunocompetent and 16 immunocompromised patients with a diagnosis based on cerebrospinal fluid (CSF) culture, sCRAG titers, and cytologic or histologic confirmation of the presence of pulmonary cryptococcosis were reviewed during the course of the study. Two thoracic radiologists reviewed chest radiographs and CT scans for morphologic characteristics and the distribution of parenchymal abnormalities, and a final reading was reached by consensus. The correlation between serial radiographs and sCRAG titers was examined in 9 immunocompetent and 10 immunocompromised patients. MEASUREMENTS: Serum or CSF cryptococcal antigen. RESULTS: The most common clinical symptom was cough, which was present in 24 patients (82.8%). Pulmonary nodules were the most frequent radiologic abnormality. Cavitation within nodules and parenchymal consolidation were significantly less common in immunocompetent patients compared to immunocompromised patients (p = 0.02 and p = 0.05, respectively). Immunocompromised patients tended to have a larger extent of pulmonary involvement than immunocompetent patients, the changes seen on their serial radiographs were more variable, and their corresponding sCRAG titers were higher (> 1:256). In the immunocompetent patients, the radiographic characteristics of lesions usually improved with a corresponding decrease in sCRAG titers over time. CONCLUSIONS: Our study suggests that pulmonary cryptococcosis usually follows a benign clinical course in immunocompetent patients. Immunocompromised patients often undergo an evolution to cavitary lesions that represent a more aggressive disease nature. Serial radiographic changes and changes in sCRAG titers reliably reflect disease progression and the response to therapy. PMID- 16478850 TI - Clinical significance of nontuberculous mycobacteria isolated from respiratory specimens in Korea. AB - STUDY OBJECTIVES: Precise epidemiologic data regarding nontuberculous mycobacterial (NTM) lung infection in many Asian countries have been relatively unavailable. In order to determine the clinical significance of NTM isolated from respiratory specimens, we reviewed medical records from all patients from whom NTM isolates were recovered within a 2-year period. MATERIALS AND METHODS: We identified all NTM isolates from respiratory specimens at the Samsung Medical Center (Seoul, South Korea) obtained from January 2002 to December 2003. We then reviewed the clinical and radiologic characteristics of the patients from whom NTM was isolated. Patients were classified as having either definite, probable, or unlikely NTM lung disease, as defined by the guidelines of both the American Thoracic Society and the British Thoracic Society. RESULTS: During the study period, 1,548 NTM isolates were recovered from 794 patients. Of these 794 patients, 131 patients (17%) were determined to have definite NTM lung disease, and 64 patients (8%) were designated as probable disease candidates. The most commonly involved organisms in the definite and probable NTM lung disease cases were Mycobacterium avium complex (n = 94, 48%) and Mycobacterium abscessus (n = 64, 33%). In 195 patients with NTM lung disease, 82 patients (42%) manifested the upper lobe cavitary form, 101 patients (52%) exhibited the nodular bronchiectatic form, and 12 patients (6%) exhibited the unclassifiable form. CONCLUSIONS: About one fourth of the patients in whom NTM was isolated from respiratory specimens were found to have clinically significant NTM lung infections. The spectrum of organisms responsible for the NTM lung disease in these Korean patients differed from those reported in other regions of the world. However, the estimates of clinical significance in this study may be underestimates due to the retrospective analysis. Some of the patients might have "true" NTM lung disease that could be diagnosed with continued evaluation and follow-up. PMID- 16478851 TI - Endobronchial ultrasonography for the quantitative assessment of bronchial mural structures in lung transplant recipients. AB - BACKGROUND: Endobronchial ultrasonography (EBUS) has been shown to be an adequate tool to visualize the laminar structure of the bronchial wall. The purpose of this study was to investigate the potential of EBUS to identify and quantitatively assess bronchial wall structures in lung transplant recipients. METHODS: EBUS was performed with a radial 20-MHz ultrasonic miniprobe in lung transplant recipients undergoing surveillance bronchoscopies. Sequential pictures were taken of the main bronchus (proximal of anastomosis) and proximal right intermedius bronchus or proximal left lower lobe bronchus (distal of anastomosis), respectively. From every localization, five slides were chosen. The quantitative assessment of the digitized pictures was done with the aid of image analysis software. In addition to the comparison of the different layers between patients with and without infection and rejection, respectively, the intraclass correlation coefficients (ICCs) of the different measurements were calculated. RESULTS: From 20 EBUS examinations performed in 10 lung transplant recipients, 200 slides were selected for quantitative assessment. A five-layer composition could be identified in all selected slides. The relative area of layer two (hypoechoic submucosal tissue) of the autologous part was significantly smaller in patients with graft rejection (p = 0.04) compared to patients without rejection, and significantly larger in patients with graft infection (p = 0.02) compared to patients without graft infection. The ICC values were calculated in 50 different slides in a subset of five consecutive patients (0.91, 0.95, 0.88, and 0.91 for layers 1, 2, 3, and 5 of the autologous and 0.70, 0.92, 0.88, and 0.84 for the allogeneic parts, respectively). CONCLUSIONS: EBUS enables to discriminate different layers of the bronchi in humans and to measure the thickness of these layers in a reproducible fashion. Therefore, EBUS may be used to investigate and quantify inflammatory alterations of bronchial wall structures in vivo. PMID- 16478852 TI - Systemic inflammatory reaction after thoracoscopic talc poudrage. AB - BACKGROUND: Recent studies have reported fever as a side effect of talc poudrage during thoracoscopic pleurodesis. However, thoracoscopy itself is likely to induce systemic inflammatory reaction, as it is an interventional procedure. The aim of the study was to investigate whether systemic inflammatory response is due to talc poudrage or to thoracoscopy. METHODS: We prospectively studied two groups of patients. The first group (18 patients) underwent thoracoscopic talc poudrage, and the second group (17 patients) underwent only diagnostic thoracoscopy. We measured body temperature, as well as WBC count and C-reactive protein (CRP) levels before the procedure (baseline), and at 24 and 48 h after the procedure. No antiinflammatory medication was permitted to be used before, during, or after the procedure. All patients had a 3-month follow-up. RESULTS: The baseline patient characteristics were similar in both groups. Temperature increased significantly in the thoracoscopic talc poudrage group (overall comparison, p = 0.005) especially at 9, 12, and 24 h after the procedure. Overall, the WBC count (p = 0.004), percentage of neutrophils (p = 0.03), and CRP levels (p < 0.0001) were significantly increased in the group of patients who underwent thoracoscopic talc poudrage. On the contrary, lymphocytes were significantly decreased (overall comparison, p = 0.01) in the thoracoscopic talc poudrage group during the same period. Mild side effects, such as pain during and after thoracoscopy and subcutaneous emphysema, were noted. No severe complication, such as infection or acute respiratory failure, was noted in either group during the hospitalization or during the follow-up period. CONCLUSION: According to our results, fever and systemic inflammatory reaction is due to talc poudrage and not to thoracoscopy. PMID- 16478853 TI - Single-center experience with 250 tunnelled pleural catheter insertions for malignant pleural effusion. AB - INTRODUCTION: Malignant pleural effusions (MPEs) are a common cause of dyspnea in patients with advanced cancer. Tunnelled pleural catheters (TPCs) can be used in patients with this condition, but the published experience with them is limited. OBJECTIVE: To describe the use of TPCs in the management of MPE in a large group of patients in a clinical setting. METHODS: Retrospective analysis of 250 sequential TPC insertions in patients with MPEs in a single center. RESULTS: Two hundred fifty TPC procedures for MPE were performed in 223 patients (19 contralateral procedures and 8 repeat ipsilateral procedures) during a 3-year period. Symptom control was complete following 97 procedures (38.8%), was partial in 125 procedures (50%), and was absent in 9 procedures (3.6%); in addition, there were 10 failed TPC insertions (4.0%) and 9 TPC insertions (3.6%) without assessment of symptoms at the 2-week follow-up visit. Spontaneous pleurodesis occurred following 103 of the 240 successful TPC procedures (42.9%) and was more frequent when < or = 20% of the hemithorax contained fluid at the 2-week follow up visit (57.2% vs 25.3%, respectively; p < 0.001). Catheters stayed in place for a median duration of 56 days. Following successful TPC placement, no further ipsilateral pleural procedures were required in 90.1% of cases. The overall median survival time following TPC insertion was 144 days. Complication rates were low and compared favorably with those seen with other treatment options. CONCLUSIONS: TPC placement is an effective method of palliation for MPE that allows outpatient management and low complication rates. The insertion of a TPC should be considered as a first-line treatment option in the management of patients with MPE. PMID- 16478854 TI - Discriminating measures and normal values for expiratory obstruction. AB - OBJECTIVES: To develop mean and 95% confidence limits for the lower limit of normal (LLN) values for forced expiratory volume in 3 s (FEV3)/FVC ratio for Latin, black, and white adults; to ascertain comparative variability of the FEV1/FVC ratio, the FEV3/FVC ratio, and forced expiratory flow, midexpiratory phase (FEF(25-75)) in never-smoking adults; to evaluate their utility in measuring the effect of smoking on airflow limitation; and to develop and use the fraction of the FVC that had not been expired during the first 3 s of the FVC (1 FEV3/FVC) to identify the growing fraction of long-time-constant lung units. DESIGN: Analysis of the Third National Health and Nutrition Examination Survey (NHANES III) database of never-smokers and current smokers. PARTICIPANTS: A total of 5,938 adult never-smokers and 3,570 current smokers from NHANES III with spirometric data meeting American Thoracic Society standards. MEASUREMENTS AND RESULTS: After establishing new databases for never-smokers and current smokers, we quantified the mean and LLN values of FEV3/FVC in never-smokers, and identified spirometric abnormalities in current smokers. When associated with older age, FEV3/FVC decreases and 1 - FEV3/FVC increases as FEV1/FVC decreases. On average, using these measurements, the condition of current smokers worsened about 20 years faster than that of never-smokers by middle age. If < 80% of the mean predicted FEF(25-75) was used to identify abnormality, over one quarter of all never-smokers would have been falsely identified as being abnormal. Using 95% confidence limits, 42% of 683 smokers with reduced FEV1/FVC and/or FEV3/FVC would have been judged as normal by FEF(25-75). CONCLUSIONS: FEV1/FVC, FEV3/FVC, and 1 FEV3/FVC characterize expiratory obstruction well. In contrast, FEF(25-75) measurements can be misleading and can cause an unacceptably large number of probable false-negative results and probable false-positive results. PMID- 16478855 TI - Diffusing capacity for nitric oxide and carbon monoxide in patients with diffuse parenchymal lung disease and pulmonary arterial hypertension. AB - BACKGROUND: The passage of carbon monoxide (CO) through the alveolocapillary membrane and into the plasma and intraerythrocytic compartments determines the diffusing capacity of the lung for CO (DLCO) as defined by the Roughton and Forster equation. On the other hand, the single-breath diffusing capacity of the lung for nitric oxide (DLNO) is thought to represent the true membrane diffusing capacity because of its very high affinity for hemoglobin (Hb) and its independence from pulmonary capillary blood volume. Therefore, the DLNO/DLCO ratio can be used to differentiate between thickened alveolocapillary membranes (both DLNO and DLCO are decreased, and the DLNO/DLCO ratio is normal) and decreased perfusion of ventilated alveoli (the DLNO less decreased than the DLCO; therefore, the DLNO/DLCO ratio is high) in patients with pulmonary disease. STUDY DESIGN: We measured the combined values of DLCO and DLNO in 41 patients with diffuse parenchymal lung disease (DPLD), 26 patients with pulmonary arterial hypertension (PAH), and 71 healthy subjects. RESULTS: The DLCO (corrected to the standard Hb value) was lowered in the DPLD group (64% of predicted) and in the PAH group (64% of predicted), and was normal in the control group (105% of predicted). The DLNO/DLCO ratio in patients with PAH (4.98) was significantly higher than that in patients with DPLD (4.56) and in healthy subjects (4.36). CONCLUSION: The DLNO/DLCO ratio is significantly higher in patients with PAH than in healthy subjects, although this ratio cannot be applied as a screening test to discriminate between patients with DPLD and PAH as the overlap between these groups is too large. PMID- 16478856 TI - Updated spirometric reference values for adult Chinese in Hong Kong and implications on clinical utilization. AB - STUDY OBJECTIVES: The accuracy of reference values of lung function is important for assessment of severity and functional impairment of respiratory diseases. The aim of the study was to establish updated prediction formulae of spirometric parameters for Hong Kong Chinese and to compare the reference values with those derived from other studies in white and Chinese subjects. DESIGN: Cross-sectional multicenter study. SETTING: Lung function laboratories of eight regional hospitals in Hong Kong. PARTICIPANTS: Subjects were recruited by random-digit dialing. One thousand one hundred seventy-six subjects who fulfilled recruitment criteria underwent spirometry. MEASUREMENTS: Spirometry was performed according to American Thoracic Society recommendations, and the technique was standardized among the eight participating lung function laboratories. RESULTS: Evaluable data of 1,089 (494 men and 595 women) healthy nonsmokers aged 18 to 80 years were analyzed. Age and height were found to be the major determinants of FEV1 and FVC, with a linear decline of height-adjusted values with age in both sexes. Spirometric values of this population have increased compared to Chinese populations of similar sex, age, and height two decades ago. Reference values derived from white populations were higher than our values by 5 to 19%, and the degree of overestimation varied with age, sex, and lung function parameter. We also demonstrated that the blanket application of correction factors for Asian populations may not be appropriate. In this study cohort, the distribution-free estimation of age-related centiles was more appropriate for the determination of lower limits of normal. CONCLUSIONS: Our findings underscore the need to use reference values based on updated data derived from local populations or those matched for ethnicity and other sociodemographic characteristics. PMID- 16478857 TI - Evaluation of thoracic tumors with 18F-fluorothymidine and 18F-fluorodeoxyglucose positron emission tomography. AB - STUDY OBJECTIVES: 18F-fluorodeoxyglucose (FDG) is the most widely used positron emission tomography (PET) imaging probe used for the diagnosis, staging, restaging, and monitoring therapy response of cancer. However, its specificity is less than ideal. A new molecular imaging probe (18F-deoxyfluorothymidine [FLT]) has been developed that might afford more specific tumor imaging. The aims of this study were as follows: (1) to compare the use of FDG-PET and FLT-PET for tumor staging, (2) to compare the degree of FDG and FLT uptake in lung lesions, and (3) to determine the correlation between PET uptake intensity and tumor cell proliferation. DESIGN: FDG-PET and FLT-PET scans were performed in 11 patients with solitary pulmonary nodules and another 11 patients with known non-small cell lung cancer (NSCLC). Tracer uptake was assessed quantitatively by standardized uptake values (SUVs). Histologic evaluation of tissue samples obtained from biopsy specimens or surgical resections served as the "gold standard." Tumor cell proliferation was assessed by Ki-67 staining. RESULTS: Pathology verification was available from 99 tissue samples in the 22 patients (29 pulmonary lesions, 66 lymph node stations, and 4 extrapulmonary lesions). Thirty-three samples (33.3%) were positive for tumor tissue (22 pulmonary, 9 lymph node stations, and 2 extrapulmonary). FDG-PET findings were false-positive in three pulmonary lesions, while FLT-PET findings were false-positive in one lesion. There were two false negative findings by FDG-PET and six false-negative findings by FLT-PET. FDG uptake of the malignant lesions was significantly higher than FLT (maximum SUV, 3.1 +/- 2.6 vs 1.6 +/- 1.2 [mean +/- SD]; p < 0.05). A significant correlation was observed between FLT uptake of pulmonary lesions and Ki-67 labeling index (r = 0.60, p = 0.02) but not for FDG uptake (r = 0.27, p = not significant). CONCLUSIONS: Compared to FDG-PET, detection of primary and metastatic NSCLC by FLT-PET is limited by the relatively low FLT uptake of the tumor tissue. Thus, FLT-PET is unlikely to provide more accurate staging information or better characterization of pulmonary nodules than FDG-PET. Nevertheless, the correlation between FLT uptake and cellular proliferation suggests that future studies should evaluate the use of FLT-PET for monitoring treatment with cytostatic anticancer drugs. PMID- 16478858 TI - Characteristics and disease activity of early interstitial lung disease in subjects with true parenchymal abnormalities in the posterior subpleural aspect of the lung. AB - STUDY OBJECTIVES: To evaluate characteristics or disease activity of early interstitial lung disease (ILD) in subjects with true parenchymal abnormalities in the posterior subpleural aspect of the lung. PATIENTS AND METHODS: This study enrolled 14 subjects with dependent densities that disappeared on helical CT obtained with the subject prone (control group) and 7 subjects with true parenchymal abnormalities that remained unchanged on prone CT image but were not detectable on chest radiographs (true abnormalities group). Pulmonary function tests and serum markers for idiopathic lung fibrosis as KL-6, surfactant protein D (SP-D), and surfactant protein A (SP-A) in the two groups were evaluated. RESULTS: In the true abnormalities group, curvilinear subpleural lines or thickened interlobular and intralobular lines were observed more frequently in the lower lung fields. Diffusing capacities of the lung for carbon monoxide (15.3 +/- 3.5 mL/min/mm Hg vs 18.8 +/- 3.7 mL/min/mm Hg, p = 0.0493) were lower, and KL 6 (607 +/- 297 U/mL vs 318 +/- 143 U/mL, p = 0.0090), SP-A (59 +/- 24 ng/mL vs 34 +/- 12 ng/mL, p = 0.0207), and SP-D (112 +/- 54 ng/mL vs 42 +/- 24 ng/mL, p = 0.0028) were higher in the true abnormalities group than in the control group (+/ SD). CONCLUSION: True parenchymal abnormalities in the posterior subpleural aspect of the lung may indicate early ILD activity. PMID- 16478859 TI - Increased serum vascular endothelial growth factor level in Churg-Strauss syndrome. AB - BACKGROUND: Churg-Strauss syndrome (CSS) is a rare form of systemic vasculitis occurring in patients with asthma and hypereosinophilia. For optimal treatment, prompt distinction of CSS from asthma is necessary; however, there are few serologic screening markers for this purpose. Vascular endothelial growth factor (VEGF), a vascular permeability factor, has been associated with other systemic vasculitis such as Wegener granulomatosis and giant-cell arteritis. OBJECTIVE: The aim of this study was to clarify the clinical value of the measurement of serum VEGF for the distinction of CSS from asthma. METHODS: We investigated serum VEGF levels in 18 CSS patients, 19 asthma patients, and 12 acute bronchitis patients. We also performed immunohistochemical analysis for VEGF. RESULTS: The serum VEGF levels of CSS patients were significantly higher than those of asthma patients and acute bronchitis patients. The sensitivity and specificity to distinguish CSS from asthma were 93.3% and 81.8%, respectively (cutoff, 600 pg/mL). Infiltrating eosinophils stained intensely positive for VEGF, and serum VEGF levels showed a significant correlation with peripheral eosinophil counts. Serum VEGF levels decreased significantly after therapy (p < 0.001). The infiltrating eosinophils in the CSS lesion stained positive for VEGF in the immunohistochemical analysis. CONCLUSION: VEGF is one of the useful screening markers for the distinction of CSS from asthma. We suggest that VEGF might be associated with the pathogenesis of CSS. PMID- 16478860 TI - Increased thromboembolic events after lung transplantation. AB - BACKGROUND: Lung transplantation is a good therapeutic option for end-stage lung disease. Data on thromboembolic complications following lung transplantation are scarce. STUDY OBJECTIVES: To evaluate the incidence of thromboembolic events following lung transplantation, and to determine their possible association with hypercoagulable state. DESIGN: Retrospective study in a single, tertiary-care, university-affiliated referral center. SUBJECTS AND METHOD: The records of 70 patients who underwent lung transplantation in our institution between September 1997 and September 2003 were reviewed for thromboembolic complications. Parameters pertaining to risk of thrombophilia were measured in the patients with thromboembolic complications. RESULTS: Thromboembolic complications developed in 6 of the 70 patients (8.6%) at 4 to 24 months after transplantation: deep vein thrombosis (DVT) in 2 patients, pulmonary embolism (PE) in 1 patient, both DVT and PE in 1 patient, and retinal vein thrombosis in 2 patients. The fibrinogen level was elevated in all six patients, and factor VIII, IX, and/or XI levels were elevated in five patients. Heterozygosity for 5 10-methylene tetrahydrofolate reductase was documented in two patients, and mutation for factor II or factor V-Leiden mutation was found in one patient. Levels of protein C and protein S and activated protein C resistance were within normal range in all patients. Four patients had mildly elevated levels of at least one antiphospholipid antibody; none had a positive lupus anticoagulant test result. Overall, all patients demonstrated abnormalities on hypercoagulability tests. CONCLUSIONS: Thromboembolic complications occur at a high rate (8.6%) in lung transplant recipients and are associated with abnormalities in hypercoagulability. The cause remains unclear. Our results should prompt a high index of suspicion for these potentially fatal complications, which would lead to early diagnosis and successful treatment. PMID- 16478861 TI - Pulmonary hemodynamic responses to brain natriuretic peptide and sildenafil in patients with pulmonary arterial hypertension. AB - STUDY OBJECTIVES: Brain natriuretic peptide (BNP) blunts hypoxic pulmonary hypertension in animal models, but its acute hemodynamic effects in patients with pulmonary arterial hypertension (PAH) are not known. The aim of this study was to determine if human B-type natriuretic peptide is a safe and efficacious pulmonary vasodilator in patients with PAH and if the pulmonary hemodynamic effects are potentiated by phosphodiesterase inhibition. DESIGN: Open-label study. SETTING: Medical ICUs of three tertiary care hospitals in New England. PATIENTS: Thirteen consecutive adult patients undergoing right-heart catheterization and a pulmonary vasodilator trial for the initial evaluation of PAH. INTERVENTIONS: Patients were administered inhaled nitric oxide (iNO), i.v. epoprostenol, and a 3-h infusion of BNP alone and 1 h after an oral dose of the phosphodiesterase-5 inhibitor sildenafil. RESULTS: iNO and sildenafil alone decreased mean pulmonary artery pressure (mPAP) without a significant fall in pulmonary vascular resistance (PVR). Epoprostenol decreased both mPAP and PVR. BNP alone had no significant effect on pulmonary hemodynamics, but the combination of sildenafil plus BNP decreased mPAP and PVR for up to 6 h after stopping BNP. The decrease in mPAP with sildenafil plus BNP (+/- SE) was greater than after 1 h of sildenafil alone (44.6 +/- 3.8 to 40.6 +/- 3.9 mm Hg, p = 0.027). An acute vasodilator response, defined as a decrease in mPAP > 10 mm Hg and end mPAP < 40 mm Hg, was seen in 0 of 8 patients with iNO, 1 of 13 patients with epoprostenol, 0 of 13 patients with BNP, and 4 of 12 patients with sildenafil plus BNP. BNP decreased mean systemic arterial pressure (5.6 +/- 2.8 mm Hg) but had no effect on cardiac output or systemic vascular resistance. CONCLUSIONS: A 3-h BNP infusion does not significantly improve pulmonary hemodynamics in most patients with PAH but is well tolerated and augments the acute pulmonary vasodilator effect of sildenafil. PMID- 16478862 TI - Normative data for pH of exhaled breath condensate. AB - INTRODUCTION: Measurement of pH is one of the simplest and most technically validated biomarkers studied in exhaled breath condensate (EBC). The pH of EBC has been found to be lower than controls in many respiratory disorders. Published data from normal control subjects have been reasonably consistent, but the data sets are not large. This study was undertaken to establish normative EBC pH reference values. PARTICIPANTS: Four hundred four healthy subjects of all ages were enrolled. INTERVENTIONS: Each participant provided a single EBC sample using a disposable collector at modest temperature so that EBC was collected as a liquid. MEASUREMENTS AND RESULTS: Samples of EBC were bubbled with argon gas to standardize for carbon dioxide, and pH was recorded with a calibrated and validated glass microelectrode on stabilization. The median EBC pH was 8.0 with interquartile (25 to 75%) range of 7.8 to 8.1. There were no differences based on age, sex, or race. The distribution is skewed, with 6.4% of EBC samples having a pH range < 7.4. CONCLUSIONS: An extensive normal data set now exists that reveals EBC pH is maintained in a modestly alkaline and tight range in subjects who consider themselves healthy. PMID- 16478863 TI - Oxidative changes of bronchoalveolar proteins in cystic fibrosis. AB - Chronic bacterial infection and severe, polymorphonuclear neutrophil-dominated endobronchial inflammation are characteristic hallmarks of cystic fibrosis (CF) lung disease. The free radicals generated can be deleterious for structure and function of many proteins. The goal of this study was to investigate the degree of oxidation of pulmonary epithelial lining fluid proteins. BAL fluid (BALF) from 55 children with CF and from 11 patients in a control group were investigated by dot-blot assay for content and by two-dimensional electrophoresis and Western blotting for the pattern of distribution of oxidized proteins. The highest level of oxidative stress, as assessed by the level of protein carbonyls, was found in patients with FEV1 < 80% of predicted or with highly elevated neutrophil counts. Compared to control subjects without lung disease, CF patients with normal lung function and CF patients with a normal neutrophil count in their BALF had significantly higher protein carbonyl levels. The extent of protein oxidation was directly related to the neutrophil granulocyte count and inversely to lung function. Our data support the hypothesis that oxidative damage of pulmonary proteins during chronic and excessive neutrophilic endobronchial inflammation may contribute to the decline of lung function in CF patients. PMID- 16478865 TI - Applied medical informatics for the chest physician: information you can use! AB - The world of applied medical informatics is changing rapidly. This is the first of a three-part series of articles on applied medical informatics that will bring the practicing chest physician up to date on the structure, function, benefits, and drawbacks of the electronic medical record and all of its components, including the virtual ICU and the daily practice of medicine. PMID- 16478864 TI - The oxyhemoglobin dissociation curve in liver cirrhosis. AB - STUDY OBJECTIVES: To trace the entire oxyhemoglobin dissociation curve (ODC) in a cohort of cirrhotic patients in stable condition who were candidates for orthotopic liver transplantation (OLT). DESIGN: Prospective cohort study. SETTING: A large academic hospital. PATIENTS AND METHODS: We traced the entire ODC in whole blood in standard conditions (pH 7.4; PCO2, 40 mm Hg; temperature, 37 degrees C) for 50 cirrhotic candidates for OLT (27 men and 23 women) and 50 age- and height-matched healthy subjects (27 men and 23 women). All subjects were nonsmokers or ex-smokers for at least 5 years. We also measured 2,3 diphosphoglycerate (2,3 DPG) in RBCs, plasma ions, and arterial blood gases in all subjects according to standard methods. Mixed venous blood was also obtained from the 50 cirrhotic patients. RESULTS: Mean ODC was the same in the two groups. However, for the cirrhotic patients, the dispersion of the PO2 values of oxygen saturation percentage (SO2%) from 20 to 80% was significantly larger (p < 0.01 to p < 0.0001). In the cirrhotic patients, the mean PO2 for half-saturation of hemoglobin (P50) was 7.11 + 0.14 mEq/L chloride (p < 0.001) plus 0.36 mEq/L inorganic phosphate (p < 0.05) plus 0.25 micromol/gram of hemoglobin (gHb) 2,3 DPG (p < 0.00002) in absolute numerical values. Sodium, potassium, and calcium, three plasma ions disturbed in cirrhotic patients, did not contribute to determine the mean P50. DISCUSSION: In patients with cirrhosis, increased dispersion of PO2 values for a given level of SO2% may be related to four factors: (1) an observed alteration of the enzymes controlling the phosphoglycerate shunt; (2) hypothyroidism, which may affect 7 to 20% of patients with primary biliary cirrhosis; (3) the type of ongoing treatment, eg, diuretics and/or propranolol; and (4) plasma ion disturbances. CONCLUSIONS: We describe the ODC by three indexes: shape, position, and an index of dispersion of the PO2 values for a given level of SO2%. In addition, when the latter is increased, we suggest that other factors than pH, temperature, carbon dioxide, and inorganic phosphates are acting on the position of the ODC. PMID- 16478866 TI - Update in the diagnosis and management of pulmonary vasculitis. AB - The term vasculitis encompasses a number of distinct clinicopathologic disease entities, each of which is characterized pathologically by cellular inflammation and destruction of the blood vessel wall, and clinically by the types and locations of the affected vessels. While multiple classification schemes have been proposed to categorize and simplify the approach to these diseases, ultimately their diagnosis rests on the identification of particular patterns of clinical, radiologic, laboratory, and pathologic features. While lung involvement is most commonly seen with the primary idiopathic, small-vessel or antineutrophil cytoplasmic antibody (ANCA)-associated vasculitides of Wegener granulomatosis, microscopic polyangiitis, and Churg-Strauss syndrome, one should remember that medium-vessel vasculitis (ie, classic polyarteritis nodosa), large-vessel vasculitis (ie, Takayasu arteritis), primary immune complex-mediated vasculitis (ie, Goodpasture syndrome), and secondary vasculitis (ie, systemic lupus erythematosus) can all affect the lung. However, for the purpose of this review, we will focus on the ANCA-associated vasculitides. PMID- 16478867 TI - Incorporating anti-IgE (omalizumab) therapy into pulmonary medicine practice: practice management implications. AB - Despite aggressive therapy, many asthma patients cannot achieve optimal control, and it is recognized that a small number of patients, generally those with severe persistent asthma, are the most difficult to control and are responsible for a large segment of the costs of asthma. These patients demonstrate a need for additional therapeutic options to achieve enhanced asthma control. Omalizumab should be considered a second-line therapy for patients with moderate-to-severe persistent allergic asthma not fully controlled on standard therapy. This article should not be considered a complete guide to incorporating this therapy into practice but serve as an introduction and a basic review of the practice management aspects of therapy for physicians practicing in the United States. PMID- 16478868 TI - Cavitating invasive pulmonary aspergillosis visualized and diagnosed by ultrathin bronchoscopy. AB - A definitive diagnosis of invasive pulmonary aspergillosis (IPA), which usually occurs in immunocompromised patients, is often difficult. We report two cases of cavitating IPA in a peripheral pulmonary region in patients who were receiving corticosteroids, in whom the cavity was successfully visualized and sampled during ultrathin bronchoscopy. Ultrathin bronchoscopy provides a new option for definitive diagnosis of cavitating IPA. PMID- 16478869 TI - Closure of a bronchopleural fistula using bronchoscopic placement of an endobronchial valve designed for the treatment of emphysema. AB - Pneumothoraces are sometimes complicated by a persistent air leak or bronchopleural fistula requiring prolonged chest tube drainage. Non-surgical treatment of persistent bronchopleural fistulas is often performed in patients who are poor surgical candidates, but the ideal method of closure is not known. Here we report closure of a persistent distal bronchopleural fistula using a one way endobronchial valve designed for the treatment of emphysema. PMID- 16478870 TI - A 35-year-old man with fever, dyspnea, and diffuse reticular opacities. PMID- 16478871 TI - A 55-year-old man with fever together with submandibular and lung masses. PMID- 16478872 TI - Correcting data for body size may confound results. PMID- 16478873 TI - CO2 can be good for you! PMID- 16478874 TI - Smoking in movies. PMID- 16478875 TI - Lung resection in the elderly. PMID- 16478876 TI - It's in the definition. PMID- 16478877 TI - It's Pneumocystis jiroveci not Pneumocystis carinii. PMID- 16478878 TI - Chest radiography and catheterization. PMID- 16478879 TI - Expression of Jak2V617F causes a polycythemia vera-like disease with associated myelofibrosis in a murine bone marrow transplant model. AB - An acquired somatic mutation, Jak2V617F, was recently discovered in most patients with polycythemia vera (PV), chronic idiopathic myelofibrosis (CIMF), and essential thrombocythemia (ET). To investigate the role of this mutation in vivo, we transplanted bone marrow (BM) transduced with a retrovirus expressing either Jak2 wild-type (wt) or Jak2V617F into lethally irradiated syngeneic recipient mice. Expression of Jak2V617F, but not Jak2wt, resulted in clinicopathologic features that closely resembled PV in humans. These included striking elevation in hemoglobin level/hematocrit, leukocytosis, megakaryocyte hyperplasia, extramedullary hematopoiesis resulting in splenomegaly, and reticulin fibrosis in the bone marrow. Histopathologic and flow cytometric analyses showed an increase in maturing myeloid lineage progenitors, although megakaryocytes showed decreased polyploidization and staining for acetylcholinesterase. In vitro analysis of primary cells showed constitutive activation of Stat5 and cytokine-independent growth of erythroid colony-forming unit (CFU-E) and erythropoietin hypersensitivity, and Southern blot analysis for retroviral integration indicated that the disease was oligoclonal. Furthermore, we observed strain-specific differences in phenotype, with Balb/c mice demonstrating markedly elevated leukocyte counts, splenomegaly, and reticulin fibrosis compared with C57Bl/6 mice. We conclude that Jak2V617F expression in bone marrow progenitors results in a PV-like syndrome with myelofibrosis and that there are strain-specific modifiers that may in part explain phenotypic pleiotropy of Jak2V617F-associated myeloproliferative disease in humans. PMID- 16478880 TI - Infants with acute lymphoblastic leukemia and a germline MLL gene are highly curable with use of chemotherapy alone: results from the Japan Infant Leukemia Study Group. AB - Although infants with acute lymphoblastic leukemia (ALL) and a germline MLL gene have a better prognosis than comparable infants with a rearranged MLL gene, their optimal therapy is controversial. In 2 consecutive studies, conducted between 1996 and 2002, we treated 22 cases of infant ALL with germline MLL using chemotherapy alone. The 5-year event-free survival rate was 95.5% with a 95% confidence interval of 86.9 to 100%. All 21 infants with precursor B-cell ALL have been in first complete remission for 3.5 to 8.8 years. Most treatment related toxicities were predictable and well tolerated, and neither secondary malignancies nor physical growth impairments have been observed. These results indicate that chemotherapy of the type described here is both safe and highly effective against infant precursor B-cell ALL with MLL in the germline configuration. PMID- 16478881 TI - Rho-mediated regulation of tight junctions during monocyte migration across the blood-brain barrier in HIV-1 encephalitis (HIVE). AB - The blood-brain barrier (BBB) is compromised during progressive HIV-1 infection, but how this occurs is incompletely understood. We studied the integrity of tight junctions (TJs) of brain microvascular endothelial cells (BMVECs) in an in vitro BBB system and in human brain tissues with HIV-1 encephalitis (HIVE). A downregulation of TJ proteins, claudin-5 and occludin, paralleled monocyte migration into the brain during HIVE. Because small G proteins (such as Rho) can play a role in BMVEC TJ assembly, an artificial BBB system explored the relationship among TJs, Rho/Rho kinase (RhoK) activation, and transendothelial monocyte migration. Coculture of monocytes with endothelial cells led to Rho activation and phosphorylation of TJ proteins. Rho and RhoK inhibitors blocked migration of infected and uninfected monocytes. The RhoK inhibitor protected BBB integrity and reversed occludin/claudin-5 phosphorylation associated with monocyte migration. BMVEC transfection with a constitutively active mutant of RhoK led to dislocation of occludin from the membrane and loss of BMVEC cell contacts. When dominant-negative RhoK-transfected BMVECs were used in BBB constructs, monocyte migration was reduced by 84%. Thus, loss of TJ integrity was associated with Rho activation caused by monocyte brain migration, suggesting that Rho/RhoK activation in BMVECs could be an underlying cause of BBB impairment during HIVE. PMID- 16478882 TI - Long-term results and competing risk analysis of the H89 trial in patients with advanced-stage Hodgkin lymphoma: a study by the Groupe d'Etude des Lymphomes de l'Adulte (GELA). AB - From 1989 to 1996, 533 eligible patients with stage IIIB/IV Hodgkin lymphoma (HL) were randomly assigned to receive 6 cycles of hybrid MOPP/ABV (mechlorethamine, vincristine, procarbazine, prednisone/Adriamycin [doxorubicin], bleomycin, vinblastine; n = 266) or ABVPP (doxorubicin, bleomycin, vinblastine, procarbazine, prednisone; n = 267). Patients in complete remission (CR) or partial response of at least 75% after 6 cycles received 2 cycles of consolidation chemotherapy (CT) (n = 208) or subtotal nodal irradiation (RT) (n = 210). A better survival probability was observed after ABVPP alone: the 10-year overall survival (OS) estimates were 90% for ABVPP x 8, 78% for MOPP/ABV x 8, 82% for MOPP/ABV with RT, and 77% for ABVPP x 6 with RT (P = .03); and the 10-year disease-free survival (DFS) estimates were 70%, 76%, 79%, and 76%, respectively (P = .09). The 10-year DFS estimates for patients treated with consolidation CT or RT were 73% and 78% (P = .07), and OS estimates were 84% and 79%, respectively (P = .29). These results showed that RT was not superior to consolidation CT after a doxorubicin-induced CR in patients with advanced HL. An analysis of competing risks identified age more than 45 years as a significant risk factor for death, relapse, and second cancers. Prospective evaluation of late adverse events may improve the management of patients with HL. PMID- 16478884 TI - Interferon-lambda-treated dendritic cells specifically induce proliferation of FOXP3-expressing suppressor T cells. AB - The lambda interferons (IFN-lambdas), also known as IL-28 and IL-29, are coexpressed with IFN-beta after Toll-like-receptor (TLR) stimulation in human monocyte-derived dendritic cells (DCs). IFN-lambda shares with type I IFNs an intracellular signaling pathway that drives the expression of a common set of genes. However, IFN-lambda signaling is initiated through a membrane receptor system distinct from that of type I IFNs. Because IFNs produced by DCs in response to TLR stimulation are critical in the differentiation and maturation of DCs, we sought to investigate whether IFN-lambda exhibits specific effects on DC differentiation. In this work, we show that DCs acquire IFN-lambda responsiveness through the expression of the specific IFN-lambda receptor chain during their differentiation from monocytes. IFN-lambda-treated DCs express high levels of major histocompatibility complex class I (MHC class I) and MHC class II but low levels of costimulatory molecules. However, they express CCR7 and acquire the ability to migrate to lymph nodes when intravenously injected into SCID/Bg mice. In mixed lymphocyte reaction (MLR) cultures, IFN-lambda-treated DCs specifically induced IL-2-dependent proliferation of a CD4(+)CD25(+)Foxp3(+) T-cell subset with contact-dependent suppressive activity on T-cell proliferation initiated by fully mature DCs. IFN-lambdas are thus able to generate tolerogenic DCs, an activity that could thwart IFN-beta functions. PMID- 16478883 TI - Allogeneic T cells induce rapid CD34+ cell differentiation into CD11c+CD86+ cells with direct and indirect antigen-presenting function. AB - Dendritic cells (DCs) derive from CD34+ cells or monocytes and stimulate alloimmune responses in transplantation. We hypothesized that the interaction between CD34+ cells and allogeneic T cells would influence the function of hematopoietic stem cells (HSCs). Cord blood (CB) CD34+ cells proliferated briskly in response to allogeneic, but not autologous, T cells when mixed with irradiated T cells for 6 days in vitro. This proliferation was significantly inhibited by an anti-HLA class II monoclonal antibody (mAb), by an anti-TNFalpha mAb, or by CTLA4 Ig. Allogeneic T cells induced the differentiation of CD34+ progenitors into cells with the morphology of dendritic monocytic precursors and characterized by the expression of HLA-DR, CD86, CD40, CD14, and CD11c, due to an endogenous release of TNFalpha. Cotransplantation of CD34+ cells with allogeneic T cells into nonobese diabetic-severe combined immunodeficiency (NOD/SCID) mice resulted in a greater engraftment of myeloid CD1c+ dendritic cells compared with cotransplantation with autologous T cells. In vitro, CD34+ cell-derived antigen presenting cells (APCs) were functionally capable of both direct and indirect presentation of alloantigens. Based on these findings, we hypothesize that in HSC transplantation the initial cross talk between allogeneic T cells and CD34+ cells may result in the increased generation of APCs that can present host alloantigens and possibly contribute to the development of graft-versus-host disease. PMID- 16478885 TI - Foxp3+CD4+CD25+ T cells control virus-specific memory T cells in chimpanzees that recovered from hepatitis C. AB - Hepatitis C virus (HCV) poses a global health problem because it readily establishes persistent infection and a vaccine is not available. CD4(+)CD25(+) T cells have been implicated in HCV persistence because their frequency is increased in the blood of HCV-infected patients and their in vitro depletion results in increased IFN-gamma production by HCV-specific T cells. Studying a well-characterized cohort of 16 chimpanzees, the sole animal model for HCV infection, we here demonstrate that the frequency of Foxp3(+)CD4(+)CD25(+) regulatory T cells (T(Regs)) and the extent of suppression was as high in spontaneously HCV-recovered chimpanzees as in persistently HCV-infected chimpanzees. Foxp3(+)CD4(+)CD25(+) T(Regs) suppressed IFN-gamma production, expansion, and activation-induced cell death of HCV-specific T cells after recovery from HCV infection and in persistent HCV infection. Thus, T(Reg) cells control HCV-specific T cells not only in persistent infection but also after recovery, where they may regulate memory T-cell responses by controlling their activation and preventing apoptosis. However, Foxp3(+)CD4(+)CD25(+) T(Reg) cells of both HCV-recovered and HCV-infected chimpanzees differed from Foxp3(+)CD4(+)CD25(+)T(Reg) cells of HCV-naive chimpanzees in increased IL-2 responsiveness and lower T-cell receptor excision circle content, implying a history of in vivo proliferation. This result suggests that HCV infection alters the population of Foxp3(+)CD4(+)CD25(+) T(Reg) cells. PMID- 16478886 TI - Phenotypic correction of von Willebrand disease type 3 blood-derived endothelial cells with lentiviral vectors expressing von Willebrand factor. AB - Von Willebrand disease (VWD) is an inherited bleeding disorder, caused by quantitative (type 1 and 3) or qualitative (type 2) defects in von Willebrand factor (VWF). Gene therapy is an appealing strategy for treatment of VWD because it is caused by a single gene defect and because VWF is secreted into the circulation, obviating the need for targeting specific organs or tissues. However, development of gene therapy for VWD has been hampered by the considerable length of the VWF cDNA (8.4 kb [kilobase]) and the inherent complexity of the VWF protein that requires extensive posttranslational processing. In this study, a gene-based approach for VWD was developed using lentiviral transduction of blood-outgrowth endothelial cells (BOECs) to express functional VWF. A lentiviral vector encoding complete human VWF was used to transduce BOECs isolated from type 3 VWD dogs resulting in high-transduction efficiencies (95.6% +/- 2.2%). Transduced VWD BOECs efficiently expressed functional vector-encoded VWF (4.6 +/- 0.4 U/24 hour per 10(6) cells), with normal binding to GPIbalpha and collagen and synthesis of a broad range of multimers resulting in phenotypic correction of these cells. These results indicate for the first time that gene therapy of type 3 VWD is feasible and that BOECs are attractive target cells for this purpose. PMID- 16478889 TI - The cover. The Cotton Exchange. PMID- 16478887 TI - Plasminogen inhibits TNFalpha-induced apoptosis in monocytes. AB - Monocytes are major mediators of inflammation, and apoptosis provides a mechanism for regulating the inflammatory response by eliminating activated macrophages. Furthermore, as a consequence of apoptosis, plasminogen binding is markedly increased on monocytoid cells. Therefore, we investigated the ability of plasminogen to modulate monocyte apoptosis. Apoptosis of monocytoid cells (human monocytes and U937 cells) was induced with either TNFalpha or cycloheximide. When apoptosis was induced in the presence of increasing concentrations of plasminogen, apoptosis was inhibited in a dose-dependent manner with full inhibition achieved at 2 microM plasminogen. Plasminogen treatment also markedly reduced internucleosomal DNA fragmentation and reduced levels of active caspase 3, caspase 8, and caspase 9 induced by TNFalpha or by cycloheximide. We examined the requirement for plasmin proteolytic activity in the cytoprotective function of plasminogen. A plasminogen active site mutant, [D(646)E]-Plg, failed to recapitulate the cytoprotective effect of wild-type plasminogen. Furthermore, antibodies against PAR1 blocked the antiapoptotic effect of plasminogen. Our results suggest that plasminogen inhibits monocyte apoptosis. The cytoprotective effect of plasminogen requires plasmin proteolytic activity and requires PAR1. Because apoptosis of monocytes plays a key role in inflammation and atherosclerosis, these results provide insight into a novel role of plasminogen in these processes. PMID- 16478888 TI - Wegener autoantigen induces maturation of dendritic cells and licenses them for Th1 priming via the protease-activated receptor-2 pathway. AB - Autoantibodies to proteinase 3 (PR3) are involved in the pathogenesis of autoimmune-mediated vasculitis in Wegener granulomatosis (WG). To address the question how the autoantigen PR3 becomes a target of adaptive immunity, we investigated the effect of PR3 on immature dendritic cells (iDCs) in patients with WG, healthy blood donors, and patients with Crohn disease (CD), another granulomatous disease. PR3 induces phenotypic and functional maturation of a fraction of blood monocyte-derived iDCs. PR3-treated DCs express high levels of CD83, a DC-restricted marker of maturation, CD80 and CD86, and HLA-DR. Furthermore, the DCs become fully competent antigen-presenting cells and can induce stimulation of PR3-specific CD4(+) T cells, which produce IFN-gamma. PR3 maturated DCs derived from WG patients induce a higher IFN-gamma response of PR3 specific CD4(+) T cells compared with patients with CD and healthy controls. The maturation of DCs mediated through PR3 was inhibited by a serine protease inhibitor, by antibodies directed against the protease-activated receptor-2 (PAR 2), and by inhibition of phospholipase C, suggesting that the interactions of PR3 with PAR-2 are involved in the induction of DC maturation. Wegener autoantigen interacts with a "gateway" receptor (PAR-2) on iDCs in vitro triggering their maturation and licenses them for a T helper 1 (Th1)-type response potentially favoring granuloma formation in WG. PMID- 16478890 TI - A piece of my mind. Errands. PMID- 16478891 TI - New oral anticoagulants show promise. PMID- 16478892 TI - Radiologists adjusting to revolution in CT. PMID- 16478893 TI - New guidelines released for managing cough. PMID- 16478894 TI - Phytoestrogens and risk of lung cancer. PMID- 16478895 TI - Beta2-adrenergic receptor genotype and survival after acute coronary syndrome. PMID- 16478896 TI - Beta2-adrenergic receptor genotype and survival after acute coronary syndrome. PMID- 16478897 TI - Internet-based continuing medical education. PMID- 16478898 TI - Pediatric research and the federal minimal risk standard. PMID- 16478899 TI - Effect of rimonabant, a cannabinoid-1 receptor blocker, on weight and cardiometabolic risk factors in overweight or obese patients: RIO-North America: a randomized controlled trial. AB - CONTEXT: Rimonabant, a selective cannabinoid-1 receptor blocker, may reduce body weight and improve cardiometabolic risk factors in patients who are overweight or obese. OBJECTIVE: To compare the efficacy and safety of rimonabant with placebo each in conjunction with diet and exercise for sustained changes in weight and cardiometabolic risk factors over 2 years. DESIGN, SETTING, AND PARTICIPANTS: Randomized, double-blind, placebo-controlled trial of 3045 obese (body mass index > or =30) or overweight (body mass index >27 and treated or untreated hypertension or dyslipidemia) adult patients at 64 US and 8 Canadian clinical research centers from August 2001 to April 2004. INTERVENTION: After a 4-week single-blind placebo plus diet (600 kcal/d deficit) run-in period, patients were randomized to receive placebo, 5 mg/d of rimonabant, or 20 mg/d of rimonabant for 1 year. Rimonabant-treated patients were rerandomized to receive placebo or continued to receive the same rimonabant dose while the placebo group continued to receive placebo during year 2. MAIN OUTCOME MEASURES: Body weight change over year 1 and prevention of weight regain during year 2. Additional efficacy measures included changes in waist circumference, plasma lipid levels, and other cardiometabolic risk factors. RESULTS: At year 1, the completion rate was 309 (51%) patients in the placebo group, 620 (51%) patients in the 5 mg of rimonabant group, and 673 (55%) patients in the 20 mg of rimonabant group. Compared with the placebo group, the 20 mg of rimonabant group produced greater mean (SEM) reductions in weight (-6.3 [0.2] kg vs -1.6 [0.2] kg; P<.001), waist circumference (-6.1 [0.2] cm vs -2.5 [0.3] cm; P<.001), and level of triglycerides (percentage change, -5.3 [1.2] vs 7.9 [2.0]; P<.001) and a greater increase in level of high-density lipoprotein cholesterol (percentage change, 12.6 [0.5] vs 5.4 [0.7]; P<.001). Patients who were switched from the 20 mg of rimonabant group to the placebo group during year 2 experienced weight regain while those who continued to receive 20 mg of rimonabant maintained their weight loss and favorable changes in cardiometabolic risk factors. Use of different imputation methods to account for the high rate of dropouts in all 3 groups yielded similar results. Rimonabant was generally well tolerated; the most common drug-related adverse event was nausea (11.2% for the 20 mg of rimonabant group vs 5.8% for the placebo group). CONCLUSIONS: In this multicenter trial, treatment with 20 mg/d of rimonabant plus diet for 2 years promoted modest but sustained reductions in weight and waist circumference and favorable changes in cardiometabolic risk factors. However, the trial was limited by a high drop-out rate and longer-term effects of the drug require further study. Clinical Trials Registration ClinicalTrials.gov Identifier: NCT00029861. PMID- 16478900 TI - Body weight and mortality among men and women in China. AB - CONTEXT: The effect of underweight and obesity on mortality has not been well characterized in Asian populations. OBJECTIVE: To examine the relationship between body mass index (BMI) and mortality in Chinese adults. DESIGN, SETTING, AND PARTICIPANTS: A prospective cohort study in a nationally representative sample of 169,871 Chinese men and women aged 40 years or older. Data on body weight and covariables were obtained at a baseline examination in 1991 using a standard protocol. Follow-up evaluation was conducted in 1999-2000, with a response rate of 93.4% (n = 158,666). MAIN OUTCOME MEASURES: Body mass index and all-cause mortality. RESULTS: After excluding those participants with missing body weight or height values, 154,736 adults were included in the analysis. After adjustment for age, sex, cigarette smoking, alcohol consumption, physical activity, education, geographic region (north vs south), and urbanization (urban vs rural), a U-shaped association between BMI and all-cause mortality was observed (P<.001). Using those participants with a BMI of 24.0 to 24.9 as the reference group, the relative risks of all-cause mortality across categories of BMI were 1.65 (95% confidence interval [CI], 1.54-1.77) for BMI less than 18.5, 1.31 (95% CI, 1.22-1.41) for BMI 18.5 to 19.9, 1.20 (95% CI, 1.11-1.29) for BMI 20.0 to 20.9, 1.12 (95% CI, 1.04-1.21) for BMI 21.0 to 21.9, 1.11 (95% CI, 1.03 1.20) for BMI 22.0 to 22.9, 1.09 (95% CI, 1.01-1.19) for BMI 23.0 to 23.9, 1.00 (95% CI, 0.92-1.08) for BMI 25.0 to 26.9, 1.15 (95% CI, 1.06-1.24) for BMI 27.0 to 29.9, and 1.29 (95% CI, 1.16-1.42) for BMI 30.0 or more. The U-shaped association existed even after excluding participants who were current or former smokers, heavy alcohol drinkers, or who had prevalent chronic illness at the baseline examination, or who died during the first 3 years of follow-up. A similar association was observed between BMI and mortality from cardiovascular disease, cancer, and other causes. CONCLUSIONS: Our results indicate that both underweight and obesity were associated with increased mortality in the Chinese adult population. Furthermore, our findings support the use of a single common recommendation for defining overweight and obesity among all racial and ethnic groups. PMID- 16478901 TI - Association of socioeconomic status with functional capacity, heart rate recovery, and all-cause mortality. AB - CONTEXT: Lower socioeconomic status (SES) confers heightened cardiovascular risk and mortality, although the mediating pathways are unclear. OBJECTIVE: To evaluate the extent to which exercise physiologic characteristics account for the association between lower SES and mortality. DESIGN, SETTING, AND PARTICIPANTS: Prospective cohort study of 30 043 consecutive patients living in 7 counties in northeast Ohio referred between 1990 and 2002 for symptom-limited stress testing for evaluation of known or suspected coronary artery disease. Follow-up for mortality continued through February 2004. MAIN OUTCOME MEASURES: Estimated functional capacity in metabolic equivalents and heart rate recovery, physiologic characteristics that are determined directly from exercise; testing and all-cause mortality during a median follow-up of 6.5 years. RESULTS: Multivariable models adjusting for demographics, insurance status, smoking status, and clinical confounders demonstrated a strong association between a composite SES score based on census block data and functional capacity (adjusted odds ratio comparing 25th with 75th percentile values, 1.72; 95% confidence interval [CI], 1.56-1.89; P<.001) as well as heart rate recovery (adjusted odds ratio comparing 25th with 75th percentile values, 1.18; 95% CI, 1.07-1.30; P<.001). There were 2174 deaths, with mortality risk increasing from 5% to 10% as SES decreased by quartile (P<.001). Cox proportional hazards models that included all confounding variables except exercise physiologic characteristics demonstrated increased mortality as SES decreased (adjusted hazard ratio comparing 25th with 75th percentile values, 1.32; 95% CI, 1.22-1.42; P<.001). After further adding functional capacity and heart rate recovery, the magnitude of this relationship was reduced (comparing 25th with 75th percentile values; adjusted hazard ratio, 1.17; 95% CI, 1.08-1.26; P<.001), with these variables explaining 47% of the association. CONCLUSIONS: Impaired functional capacity and abnormal heart rate recovery were strongly associated with lower SES and accounted for a major proportion of the correlation between SES and mortality. Efforts to modify these clinical features among patients with low SES may narrow disparities in mortality. PMID- 16478902 TI - Intravenous morphine and topical tetracaine for treatment of pain in [corrected] neonates undergoing central line placement. AB - CONTEXT: There is limited evidence of the analgesic effectiveness of opioid analgesia or topical anesthesia during central line placement in neonates, and there are no previous studies of their relative effectiveness. OBJECTIVE: To determine the effectiveness and safety of topical tetracaine, intravenous morphine, or tetracaine plus morphine for alleviating pain in ventilated neonates during central line placement. DESIGN, SETTING, AND PARTICIPANTS: Randomized, double-blind, controlled trial enrolling 132 ventilated neonates (mean gestational age, 30.6 [SD, 4.6] weeks at study entry) and conducted between October 2000 and July 2005 in 2 neonatal intensive care units in Toronto, Ontario. INTERVENTIONS: Prior to central line insertion, neonates were randomly assigned to receive tetracaine (n = 42), morphine (n = 38), or both (n = 31); a separate nonrandomized group of 21 neonates receiving neither tetracaine nor morphine was used as a control group. MAIN OUTCOME MEASURES: The primary outcome measure was a pain score for the proportion of time neonates displayed facial grimacing (brow bulge) during different phases of the procedure (skin preparation, needle puncture, and recovery). In randomized neonates, safety assessments included blood pressure, ventilatory support, and local skin reactions. RESULTS: Compared with no treatment, pain scores were lower in the morphine and tetracaine-morphine groups during skin preparation (mean difference, -0.22; 95% confidence interval [CI], -0.4 to -0.04; P = .02 and -0.29; 95% CI, 0.49 to -0.09; P = .01, respectively), and needle puncture (mean difference, 0.35; 95% CI, -0.57 to -0.13; P = .003 and -0.47; 95% CI, -0.71 to -0.24; P<.001, respectively), but pain scores did not differ statistically for tetracaine alone vs no treatment. Pain scores were lower for morphine and tetracaine-morphine vs tetracaine during the skin preparation phase and for tetracaine-morphine vs tetracaine during needle puncture. Compared with neonates without morphine, morphine-treated neonates required larger increases in ventilation rate in the first 12 hours (mean difference, 3.9/min; 95% CI, 1.3-6.5/min; P = .003). Local skin reactions occurred in 30% of neonates given tetracaine vs 0% for morphine (risk difference, 0.30; 95% CI, 0.19-0.41; P<.001). CONCLUSION: In this study of ventilated neonates undergoing central line placement, morphine and tetracaine plus morphine provided superior analgesia to tetracaine; however, morphine caused respiratory depression and tetracaine caused erythema.Clinical Trials Registration ClinicalTrials.gov Identifier: NCT00213200. PMID- 16478903 TI - Development and validation of a prognostic index for 4-year mortality in older adults. AB - CONTEXT: Both comorbid conditions and functional measures predict mortality in older adults, but few prognostic indexes combine both classes of predictors. Combining easily obtained measures into an accurate predictive model could be useful to clinicians advising patients, as well as policy makers and epidemiologists interested in risk adjustment. OBJECTIVE: To develop and validate a prognostic index for 4-year mortality using information that can be obtained from patient report. DESIGN, SETTING, AND PARTICIPANTS: Using the 1998 wave of the Health and Retirement Study (HRS), a population-based study of community dwelling US adults older than 50 years, we developed the prognostic index from 11,701 individuals and validated the index with 8009. Individuals were asked about their demographic characteristics, whether they had specific diseases, and whether they had difficulty with a series of functional measures. We identified variables independently associated with mortality and weighted the variables to create a risk index. MAIN OUTCOME MEASURE: Death by December 31, 2002. RESULTS: The overall response rate was 81%. During the 4-year follow-up, there were 1361 deaths (12%) in the development cohort and 1072 deaths (13%) in the validation cohort. Twelve independent predictors of mortality were identified: 2 demographic variables (age: 60-64 years, 1 point; 65-69 years, 2 points; 70-74 years, 3 points; 75-79 years, 4 points; 80-84 years, 5 points, >85 years, 7 points and male sex, 2 points), 6 comorbid conditions (diabetes, 1 point; cancer, 2 points; lung disease, 2 points; heart failure, 2 points; current tobacco use, 2 points; and body mass index <25, 1 point), and difficulty with 4 functional variables (bathing, 2 points; walking several blocks, 2 points; managing money, 2 points, and pushing large objects, 1 point. Scores on the risk index were strongly associated with 4-year mortality in the validation cohort, with 0 to 5 points predicting a less than 4% risk, 6 to 9 points predicting a 15% risk, 10 to 13 points predicting a 42% risk, and 14 or more points predicting a 64% risk. The risk index showed excellent discrimination with a cstatistic of 0.84 in the development cohort and 0.82 in the validation cohort. CONCLUSION: This prognostic index, incorporating age, sex, self-reported comorbid conditions, and functional measures, accurately stratifies community-dwelling older adults into groups at varying risk of mortality. PMID- 16478904 TI - Implantable cardioverter-defibrillators: expanding indications and technologies. AB - CONTEXT: Sudden cardiac death (SCD) is a major challenge facing contemporary cardiology. For an increasing number of patients, the current standard of care for the treatment and prevention of SCD is the implantable cardioverter defibrillator (ICD). Since its introduction, there have been numerous advances in ICD technology, and indications for its use have expanded greatly in the past year. OBJECTIVE: To highlight the evolving indications for and the numerous advances in ICD technology, with emphasis on primary and secondary prophylaxis of SCD. EVIDENCE ACQUISITION: Electronic literature search of the Pubmed and MEDLINE databases from January 1996 to July 2005, using the Medical Subject Heading implantable defibrillator. Abstracts and titles were reviewed to identify English language randomized controlled trials that included an ICD group and a non-ICD group and that had end points of all-cause mortality, cardiac death, and/or arrhythmic mortality as the main outcome. A further MEDLINE search was conducted to identify randomized controlled trials of cardiac resynchronization therapy (CRT) with a CRT and a non-CRT group (including both mortality and other end points). Other studies were included that clarify aspects of device function and other relevant issues. A total of 22 trials were identified. EVIDENCE SYNTHESIS: ICD implantation improves survival in patients with a history of life-threatening ventricular arrhythmia. More recent evidence shows that ICD implantation also improves survival as primary prophylaxis against SCD in patients at high risk for ventricular arrhythmias, including those with left ventricular ejection fraction (LVEF) of 35% or less and New York Heart Association class II or III heart failure and those with a history of myocardial infarction and LVEF of 30% or less. Cardiac resynchronization improves symptoms, quality of life, and survival for patients with advanced heart failure and intraventricular conduction delays and ventricular dyssynchrony. CONCLUSIONS: ICDs have been shown to improve survival as both primary and secondary prophylaxis in an expanding population of patients. Ongoing ICD research may continue to delineate groups with survival benefit from ICDs, and the use and indications of these devices in clinical practice will continue to expand. PMID- 16478905 TI - Law enforcement interviews of hospital patients: a conundrum for clinicians. PMID- 16478906 TI - Obesity research--limitations of methods, measurements, and medications. PMID- 16478914 TI - JAMA patient page. The metabolic syndrome. PMID- 16478915 TI - Chemokines, chemokine receptors, and cancer metastasis. AB - It is clear from large clinical studies that selected chemokine receptors are often up-regulated in a large number of common human cancers, including those of the breast, lung, prostate, colon, and melanoma. Chemokine receptors and their corresponding chemokine ligands have been demonstrated to play a number of nonredundant roles in cancer metastasis to vital organs as well as regional lymph nodes, the most frequent site of cancer metastasis. Chemokine receptors may potentially facilitate tumor dissemination at several key steps of metastasis, including adherence of tumor cells to endothelium, extravasation from blood vessels, metastatic colonization, angiogenesis, proliferation, and protection from the host response via activation of key survival pathways such as phosphatidylinositol-3 kinase and Akt. It is interesting that many of these roles are reminiscent of their functions in leukocyte and stem cell trafficking. Lastly, we discuss therapeutic applications for chemokine receptor antagonists in cancer therapy. PMID- 16478916 TI - Tissue-specific expression of estrogen receptors and their role in the regulation of neutrophil infiltration in various organs following trauma-hemorrhage. AB - Although 17beta-estradiol (E2) administration after trauma-hemorrhage (T-H) reduces tissue neutrophil sequestration in male rodents, it remains unknown which of the estrogen receptor (ER) subtypes mediates this effect and whether the same ER subtype is involved in all the tissues. We hypothesized that the salutary effects of E2 on attenuation of neutrophil accumulation following T-H are tissue and receptor subtype-specific. Male Sprague-Dawley rats underwent sham operation or T-H (mean blood pressure, 40 mmHg for 90 min and then resuscitation). E2 (50 microg/kg), ER-alpha agonist propyl pyrazole triol (PPT; 5 microg/kg), ER-beta agonist diarylpropiolnitrile (DPN; 5 microg/kg), or vehicle (10% dimethyl sulfoxide) was administered subcutaneously during resuscitation. Twenty-four hours thereafter, tissue myeloperoxidase (MPO) activity (a marker of neutrophil sequestration), cytokine-induced neutrophil chemoattractant (CINC)-1, CINC-3, and intercellular adhesion molecule (ICAM)-1 levels in the liver, intestine, and lung were measured (n = 6 rats/group). ER-alpha and ER-beta mRNA levels in sham operated rats were also determined. T-H increased MPO activity, CINC-1, CINC-3, and ICAM-1 levels in the liver, intestine, and lung. These parameters were improved significantly in rats receiving E2 after T-H. Administration of the ER alpha agonist PPT but not the ER-beta agonist DPN improved the measured parameters in the liver. In contrast, DPN but not PPT significantly improved these parameters in the lung. In the intestine, ER subtype specificity was not observed. ER-alpha mRNA expression was highest in the liver, whereas ER-beta mRNA expression was greatest in the lung. Thus, the salutary effects of E2 administration on tissue neutrophil sequestration following T-H are receptor subtype and tissue-specific. PMID- 16478917 TI - The long pentraxin PTX3 as a link among innate immunity, inflammation, and female fertility. AB - The long pentraxin 3 (PTX3) is member of a complex superfamily of multifunctional proteins characterized by a cyclic multimeric structure. PTX3 is highly conserved in evolution and is produced by innate-immunity cells in response to proinflammatory signals and Toll-like receptor engagement. PTX3 plays complex, nonredundant functions in vivo, acting as a predecessor of antibodies, recognizing microbes, activating complement, facilitating pathogen recognition by phagocytes, and hence, playing a nonredundant role in resistance against selected pathogens. In addition, PTX3 is essential in female fertility by acting as a nodal point for the assembly of the cumulus oophorus hyaluronan-rich extracellular matrix. Thus, the prototypic long pentraxin PTX3 is a multifunctional, soluble pattern recognition receptor acting as a nonredundant component of the humoral arm of innate immunity and involved in matrix deposition and female fertility. PMID- 16478918 TI - Regulation of phagocyte lifespan in the lung during bacterial infection. AB - The innate-immune response to infection is critically dependent on the antimicrobial actions of macrophages and neutrophils. Host and pathogen have evolved strategies to regulate immune-cell antimicrobial functions via alterations in cell death. Modulation of phagocyte death by bacteria is an important pathogenic mechanism. Host benefits of phagocyte apoptosis also exist, and understanding the mechanisms and consequences of apoptosis is essential before we can devise strategies to modulate this element of the innate-immune response to the host's benefit. This is of particular importance in an organ such as the lung, in which the balance between the need to recruit phagocytes to maintain bacterial sterility and the requirement to clear recruited cells from the alveolar units to preserve physiologic gas exchange must be finely tuned to ensure survival during bacterial infection. Apoptosis clearly plays a critical role in reconciling these physiological requirements. PMID- 16478919 TI - Matrix metalloproteinase-9 modulates intestinal injury in rats with transmural colitis. AB - Proteolysis and degradation of extracellular matrix by metalloproteinases (MMPs) may contribute to intestinal injury in inflammatory bowel disease. In the present study, we investigated the pathogenic role of gelatinases (MMP-9 and MMP-2) on transmural colonic injury in a rat model of chronic colitis, which was induced by intracolonic instillation of trinitrobenzene sulfonic acid (TNBS). The activity and expression of MMP-2 and MMP-9 were measured in colonic tissue and peripheral neutrophils by fluorescence, zymography, Western blot, or immunohistochemistry at different time-points. Furthermore, myeloperoxidase content in colonic homogenates was analyzed to evaluate inflammation. Finally, morphological changes were assessed following early or delayed administration of CGS-27023-A, a synthetic inhibitor of MMPs. We found that the induction of colitis led to a significant up-regulation in tissue gelatinase concentration, whereas no changes in collagenase activity were observed. In addition, up-regulation of pro-MMP-9, but not of pro-MMP-2, was found on Days 7 and 10 following the induction of colitis. Furthermore, transmural MMP-9 was detected by immunofluorescent staining in the inflamed tissue. Consistent with tissue samples, neutrophils from colitic rats showed a significantly increased activity of pro-MMP-9. Finally, early but not delayed treatment with CGS-27023-A attenuated colonic mucosal injury in rats with TNBS-induced colitis. In conclusion, up-regulation of MMP-9 in peripheral and colonic neutrophils modulates transmural colonic injury in rats with TNBS induced colitis. PMID- 16478920 TI - Ethanol affects the generation, cosignaling molecule expression, and function of plasmacytoid and myeloid dendritic cell subsets in vitro and in vivo. AB - The influence of ethanol (EtOH) on multiple dendritic cell (DC) subsets, in the steady state or following their mobilization in vivo, has not been characterized. Herein, generation of mouse bone marrow-derived DC (BMDC) in response to fms-like tyrosine kinase 3 ligand was inhibited by physiologically relevant concentrations of EtOH with selective suppression of plasmacytoid (p)DC. EtOH reduced surface expression of costimulatory molecules (CD40, CD80, CD86) but not that of coinhibitory CD274 (B7-H1) on resting or CpG-stimulated DC subsets. Interleukin (IL)-12p70 production by activated DC was impaired. Consistent with these findings, EtOH-exposed BMDC exhibited a reduced capacity to induce naive, allogeneic T cell proliferation and impaired ability to prime T cells in vivo. DC subsets freshly isolated from EtOH-fed mice were also examined. Liver DC, inherently immature and resistant to maturation, exhibited little change in their low surface cosignaling molecule expression, whereas splenic DC showed reduced expression of surface costimulatory molecules in response to CpG stimulation in vivo. These splenic DC elicited reduced naive, allogeneic T cell proliferation in vitro, and the stimulatory capacity of resting but not CpG-activated liver DC was reduced by chronic EtOH administration. T cells from animals primed with EtOH exposed DC produced elevated levels of IL-10 following ex vivo challenge with donor alloantigen. Thus, EtOH impairs cytokine-driven differentiation and function of myeloid DC and pDC in vitro. Hepatic DC from chronic EtOH-fed mice are less affected than splenic DC, which exhibit impaired functional maturation following CpG stimulation. These results indicate a potential mechanism by which alcohol consumption is associated with immunosuppression. PMID- 16478921 TI - Clonal restriction of the expansion of antigen-specific CD8+ memory T cells by transforming growth factor-{beta}. AB - Recent evidence showed that transforming growth factor-beta (TGF-beta) regulates the global expansion of CD8+ T cells, which are CD44hi, a marker for memory cells. However, it is not clear whether this regulatory mechanism also applies to the antigen-specific CD8+ memory cells. By using a murine mixed lymphocyte culture (MLC) model, we examined the effect of TGF-beta on antigen-specific CD8+ memory cells [cytotoxic T lymphocyte (CTL)]. We found that the secondary CTL response in CD8+ memory cells from untreated MLC was not affected by TGF-beta but augmented by interleukin (IL)-2, whereas the CD8+ memory cells from TGF-beta pretreated MLC (MLC-TGF-beta) failed to mount a significant, secondary CTL response, even when IL-2 was added. In exploring this dichotomy, in combination with flow cytometry analysis, we found that prolonged exposure to TGF-beta reduces the CTL activity in CD8+ memory cells. The increase by IL-2 and the reduction by TGF-beta of the CTL responses were clonal-specific. TGF-beta did not affect the CTL response to a third-party antigen or polyclonal T cell activation. Experiments performed with transgenic 2C cells gave similar results. Cell-cycle study performed with adoptive transfer of the cell tracker-labeled MLC cells revealed that the in vivo expansion of CD8+ memory cells from MLC-TGF-beta was restricted severely, and the restriction was clonal-specific, thus offering direct evidence to show that TGF-beta induces clonal restriction of CD8+ memory cell expansion. PMID- 16478922 TI - Conditional up-regulation of IL-2 production by p38 MAPK inactivation is mediated by increased Erk1/2 activity. AB - The p38 mitogen-activated protein kinase regulates many cellular processes in almost all eukaryotic cell types. In T cells, p38 was shown to regulate thymic development and cytokine production. Here, the role of p38 on interleukin-2 (IL 2) production by human peripheral blood CD4+ T cells was examined. When T cells were stimulated under weak stimulation conditions, pharmaceutical and molecular p38 inhibitors induced a dramatic increase of IL-2 production. In contrast, IL-2 levels were not affected significantly when strong stimulation was provided to T cells. The increase in IL-2 production, following p38 inhibition, was associated with a strong up-regulation of extracellular signal-regulated kinase (Erk)1/2 activity. Furthermore the Erk inhibitor U0126 was able to counteract the effect of p38 inhibition on IL-2 production, supporting the conclusion that p38 mediates its effect through Erk. These results suggest that the p38 kinase, through its ability to control Erk activation levels, acts as a gatekeeper, which prevents inappropriate IL-2 production. Also, the finding that p38 acts in a strength-of stimulation-dependent way provides an explanation for previously reported, contradictory results regarding the role of this kinase in IL-2 expression. PMID- 16478924 TI - Why dentists should be called oral physicians now. PMID- 16478925 TI - A framework for service-learning in dental education. AB - Service-learning has become an important component of higher education. Integrating service-learning into dental and dental hygiene curricula will foster graduates who are better prepared to work effectively among diverse populations and to function dynamically in the health policy arena. Although the phrase is familiar to dental educators, there is not a consistent understanding of what comprises this pedagogy. This article offers a framework for service-learning in dental education and describes ten components that characterize true service learning. This framework can provide a common understanding of this form of experiential education that brings community engagement and educational objectives together. More effective programs can be built around a shared understanding of the characteristics and goals of service-learning in dental education. PMID- 16478926 TI - Students' perceptions of effective learning experiences in dental school: a qualitative study using a critical incident technique. AB - Students' views of their educational experience can be an important source of information for curriculum assessment. Although quantitative methods, particularly surveys, are frequently used to gather such data, fewer studies have employed qualitative methods to examine students' dental education experiences. The purpose of this study is to explore characteristics of effective learning experiences in dental school using a qualitative method. All third-year (seventy) and fourth-year (seventy) dental students enrolled in one midwestern dental school were invited to participate. Fifty-three dental students (thirty-five male and eighteen female; thirty-two third-year and twenty-one fourth-year) were interviewed using a critical incident interview technique. Each student was asked to describe a specific, particularly effective learning incident that he or she had experienced in dental school and a specific, particularly ineffective learning incident, for comparison. Each interview was audiotaped. Students were assured that only the interviewer and one additional researcher would have access to the tapes. Data analysis resulted in identification of key themes in the data describing characteristics of effective learning experiences. The following characteristics of effective learning experiences were identified: 1) instructor characteristics (personal qualities, "checking-in" with students, and an interactive style); 2) characteristics of the learning process (focus on the "big picture," modeling and demonstrations, opportunities to apply new knowledge, high quality feedback, focus, specificity and relevance, and peer interactions); and 3) learning environment (culture of the learning environment, technology). Common themes emerged across a wide variety of learning incidents. Although additional research is needed, the characteristics of effective learning experiences identified in this study may have implications for individual course design and for the dental school curriculum as a whole. PMID- 16478928 TI - Estimating transfer of learning for self-instructional packages across dental schools. AB - The most common topic of research in dental education is assessing the effectiveness of self-instructional units in various formats compared to lectures covering the same material. Generally, these studies are of high methodological quality and reveal mixed results or results slightly favoring self-instruction. All such studies, save one, have been conducted in the context of various single schools, thus confounding the effects of self-instructional format with factors particular to schools and their students. A reanalysis, using Cronbach's generalizability analysis, was performed on a study in the literature that was conducted at six schools and measured student aptitude. The reanalysis found that the largest source of variance on immediate post-test quizzes for knowledge following a three-hour unit on disturbances in tooth development was the school at which the study was conducted (24 percent), followed by student aptitude measured by DAT score (20 percent). Difference in format among lecture, booklet, and audiotape presentations accounted for 5 percent of the variance. This reanalysis demonstrated that statistically significant results from rigorous experimental designs can overrepresent what is revealed by such research. The context-specificity of educational innovations may be underestimated because few studies are replicated across schools. Studies conducted as single schools, regardless of their methodological rigor, fail to address issues associated with potential transfer of findings to other schools. PMID- 16478927 TI - A chairside aid for shared decision making in dentistry: a randomized controlled trial. AB - The concept of shared decision making (SDM) is an important emerging trend in clinical medicine but has received little or no attention in the dental literature. Decision aids can play a useful role in SDM by helping patients and clinicians choose among reasonable alternative treatment options. The purpose of this study was to develop and test an Endodontic Decision Board (EndoDB) for chairside use to help clarify treatment alternatives, benefits, risks, prognosis, and costs when root canal therapy or extraction of a tooth was indicated. The hypothesis was that the use of the EndoDB would lead to improved patient knowledge, greater satisfaction with the decision-making process, and no difference in anxiety when compared to the standard discussion and informed consent process (usual care). The EndoDB was tested in a randomized controlled trial in a postgraduate endodontics clinic. After treatment discussion, a brief questionnaire was completed by the patient to measure knowledge, satisfaction, and anxiety. Patients in the EndoDB group (n=32) demonstrated a small, but statistically significant, increase in knowledge (t-test; difference=+0.37; p=0.03) compared to the usual care group (n=35). There was no difference between groups in the measures of satisfaction or anxiety (Mann-Whitney U-test; p>0.05). Decision aids may emerge as a useful tool to facilitate SDM and evidence-based clinical practice. PMID- 16478929 TI - Accuracy and consistency of radiographic interpretation among clinical instructors using two viewing systems. AB - Accurate and consistent radiographic interpretation among clinical instructors is needed for assessment of teaching, student performance, and patient care. The purpose of this investigation was to determine if the method of radiographic viewing affects accuracy and consistency of instructors' determinations of bone loss. Forty-one clinicians who provide instruction in a dental school clinical teaching program (including periodontists, general dentists, periodontal graduate students, and dental hygienists) quantified bone loss for up to twenty-five teeth into four descriptive categories using a view box for plain film viewing or a projection system for digitized image viewing. Ratings were compared to the correct category as determined by direct measurement using the Schei ruler. Agreement with the correct choice for the view box and projection system was 70.2 percent and 64.5 percent, respectively. The mean difference was better for a projection system due to small rater error by graduate students. Projection system ratings were slightly less consistent than view box ratings. Dental hygiene faculty ratings were the most consistent but least accurate. Although the projection system resulted in slightly reduced accuracy and consistency among instructors, training sessions utilizing a single method for projecting digitized radiographic images have their advantages and may positively influence dental education and patient care by enhancing accuracy and consistency of radiographic interpretation among instructors. PMID- 16478930 TI - Longitudinal trends in the use of individualized radiographic examinations at dental schools in the United States and Canada. AB - In the spring of 2002, a mail survey was conducted to determine the use of individualized radiographic examinations (selective radiography) for comprehensive care patients at all sixty-four U.S. and Canadian dental schools. Results from previous surveys were used to evaluate the long-term longitudinal trends. Among sixty-two schools (97 percent response rate), selective radiography was used by 34 percent of schools for dentulous adult patients, by 100 percent for edentulous adults, and by 28 percent for children. Having a credentialed chief of service increased the likelihood that selective radiography would be used for dentulous adults (odd ratio[OR]=2.36) and for children (OR=2.33). Selective radiography for dentulous adults increased from 2 percent of schools in 1977 to 36 percent in 1997 and leveled off thereafter. Between 1987 and 2002, selective radiography for edentulous adults was used at nearly all schools (96 100 percent) and for children at about a quarter of dental schools (22-28 percent). Among the sixty-one schools for which there are complete data since 1987, the continuous use of routine radiography was most common (39 percent of schools) for dentulous adult patients, whereas the continuous use of selective radiography was uncommon (7 percent). PMID- 16478931 TI - Sociodemographic backgrounds and career decisions of Australian and New Zealand dental students. AB - This article describes the sociodemographic and career selection characteristics of dental students in Australia and New Zealand. A total of 672 dental students participated in the study. The survey covered age, language proficiency, type of school attended, place of residence, parental occupation, and level of education. The respondents had an average age of twenty-two years, with a range of eighteen to fifty. Fifty-six percent of respondents were female, and approximately half had completed secondary education in private schools with 44.3 percent having finished in public schools. The majority of students lived with their parents, with only a few respondents reporting a rural home address (6.8 percent). The majority of students (65.3 percent) had placed dentistry as their first career choice and had most likely made the decision after leaving high school or near the end of high school (81.4 percent), with self-motivation being the major influence on their decision. This study provides a description of the sociodemographic profile of Australian and New Zealand dental students and provides a better understanding of career decision issues. It also highlights areas for further investigation and management by educational institutions and public policy. PMID- 16478932 TI - Oral health attitudes, knowledge, and behavior among school children in North Jordan. AB - The aim of this study was to assess the knowledge, attitude, and behavior of school children towards oral health and dental care as well as to evaluate the factors that determine these variables. School children (n=557) of an average age of 13.5 years attending public schools in North Jordan were recruited into this study. The subjects completed a questionnaire that aimed to evaluate young school children's behavior, knowledge, and perception of their oral health and dental treatment. The participants' oral hygiene habits (such as tooth brushing) were found to be irregular, and parents' role in the oral hygiene habits of their children was limited. The study population showed higher awareness of caries than periodontal conditions. Irregular visits to the dentist were found to be common, and toothache was the major driving factor for dental visits. Children had positive attitudes toward their dentists; nevertheless, they indicated that they feared dental treatment. The children in this study also recognized the importance of oral health to the well-being of the rest of the body. Parents were not proactive in making sure that their children received regular dental care. Parents' knowledge and attitudes about the importance of oral health care and their fears about dental treatment influenced their children's dental care. The results of this study indicate that children's and parents' attitudes toward oral health and dental care need to be improved. Comprehensive oral health educational programs for both children and their parents are required to achieve this goal. PMID- 16478933 TI - Dental school vacant budgeted faculty positions: academic year 2004-05. AB - The number of vacant budgeted faculty positions in dental schools has continued to decline, dropping from 296 in 2003-04 to 275 in 2004-05. The number of lost positions declined to twenty-five, from 147 lost positions reported in 2003-04. While the average number of vacancies per dental school was just under five, three-quarters of these vacancies were considered usual and normal to the operation of the dental school. Based on ADEA's annual survey of dental educators, there was approximately a 9 percent faculty turnover between 2003-04 and 2004-05, and according to the 2003-04 and 2004-05 surveys of vacant faculty positions, it is taking longer to fill vacant positions. The greatest challenges influencing the ability of a school to fill a vacancy were salary/budget limitations and lack of response to a position announcement. Still, between 2003 04 and 2004-05, the number of dental school faculty increased from 11,348 to 11,715, including 4,736 full-time, 5,097 part-time, and 1,791 volunteer faculty members. Employment status was not reported for ninety-one individuals. Private practice remains the primary reason for faculty separations and the source of new faculty. In addition, nearly one in four new faculty members entered dental education directly following graduation from a dental or postdoctoral education program. While it may take longer to fill positions and it has become more difficult to fill some vacancies, overall, dental school deans indicated that the number of vacancies was not affecting the quality of dental education. However, between anticipated faculty retirements and current levels of faculty turnover, continued support for and development of faculty recruitment and retention programs remains essential to maintaining a quality dental education workforce. PMID- 16478934 TI - Continued vigilance: enhancing diversity in dental education. PMID- 16478935 TI - Arabidopsis jasmonate signaling pathway. AB - Jasmonates control defense gene expression and male fertility in the model plant Arabidopsis thaliana. In both cases, the involvement of the jasmonate pathway is complex, involving large-scale transcriptional reprogramming. Additionally, jasmonate signaling is hard-wired into the auxin, ethylene, and salicylate signal networks, all of which are under intense investigation in Arabidopsis. In male fertility, jasmonic acid (JA) is the essential signal intervening both at the level of anther elongation and in pollen dehiscense. A number of genes potentially involved in jasmonate-dependent anther elongation have recently been discovered. In the case of defense, at least two jasmonates, JA and its precursor 12-oxo-phytodienoic acid (OPDA), are necessary for the fine-tuning of defense gene expression in response to various microbial pathogens and arthropod herbivores. However, only OPDA is required for full resistance to some insects and fungi. Other jasmonates probably affect yet more physiological responses. A series of breakthroughs have identified the SKP/CULLIN/F-BOX (SCF), CORONATINE INSENSITIVE (COI1) complex, acting together with the CONSTITUTIVE PHOTOMORPHOGENIC 9 (COP9) signalosome, as central regulatory components of jasmonate signaling in Arabidopsis. The studies, mostly involving mutational approaches, have paved the way for suppressor screens that are expected to further extend our knowledge of jasmonate signaling. When these and other new mutants affecting jasmonate signaling are characterized, new nodes will be added to the Arabidopsis Jasmonate Signaling Pathway Connections Map, and the lists of target genes regulated by jasmonates in Arabidopsis will be expanded. PMID- 16478936 TI - Jasmonate signaling pathway. AB - Jasmonates in plants are cyclic fatty acid-derived regulators structurally similar to prostaglandins in metazoans. These chemicals mediate many of plants' transcriptional responses to wounding and pathogenesis by acting as potent regulators for the expression of numerous frontline immune response genes, including those for defensins and antifungal proteins. Additionally, the pathway is critical for fertility. Ongoing genetic screens and protein-protein interaction assays are identifying components of the canonical jasmonate signaling pathway. A massive molecular machine, based on two multiprotein complexes, SCF(COI1) and the COP9 signalosome (CNS), plays a central role in jasmonate signaling. This machine functions in vivo as a ubiquitin ligase complex, probably targeting regulatory proteins, some of which are expected to be transcriptional repressors. Some defense-related mediators, notably salicylic acid, antagonize jasmonates in controlling the expression of many genes. In Arabidopsis, NONEXPRESSOR OF PR GENES (NPR1) mediates part of this interaction, with another layer of control provided further downstream by the mitogen activated protein kinase (MAPK) homolog MPK4. Numerous other interpathway connections influence the jasmonate pathway. Insights from Arabidopsis have shown that an allele of the auxin signaling gene AXR1, for example, reduces the sensitivity of plants to jasmonate. APETALA2 (AP2)-domain transcription factors, such as ETHYLENE RESPONSE FACTOR 1 (ERF1), link the jasmonate pathway to the ethylene signaling pathway. As progress in characterizing several new mutants (some of which are hypersensitive to jasmonic acid) augments our understanding of jasmonate signaling, the Connections Map will be updated to include this new information. PMID- 16478937 TI - Jasmonate biochemical pathway. AB - Plants possess an interrelated family of potent fatty acid-derived regulators-the jasmonates. These compounds, which play roles in both defense and development, are derived from tri-unsaturated fatty acids [alpha-linolenic acid (18:3) or 7Z,10Z,13Z-hexadecatrienoic acid (16:3)]. The lipoxygenase-catalyzed addition of molecular oxygen to alpha-linolenic acid initiates jasmonate synthesis by providing a 13-hydroperoxide substrate for the formation of an unstable allene oxide that is then subject to enzyme-guided cyclization to produce 12-oxo phytodienoic acid (OPDA). OPDA has several fates, including esterification into plastid lipids or transformation into the 12-carbon co-regulator jasmonic acid (JA). JA, the best-characterized member of the family, regulates both male and female fertility (depending on the plant species) and is an important mediator of defense gene expression. JA is itself a substrate for further diverse modifications. Genetic dissection of the pathway is revealing how the different jasmonates modulate different physiological processes. Each new family member that is discovered provides another key to understanding the fine control of gene expression in immune responses, in the initiation and maintenance of long distance signal transfer in response to wounding, in the regulation of fertility, and in the turnover, inactivation, and sequestration of jasmonates, among other processes. The Jasmonate Biochemical Pathway provides an overview of the growing jasmonate family, and new members will be included in future versions of the Connections Map. PMID- 16478938 TI - Focus issue: plant communication. AB - This week's issues of Science and Science's STKE focus on plant signaling with an emphasis on volatile organic compounds. Science emphasizes the chemistry of and information encoded by plant volatiles and how plants, plant pathogens, or humans utilize this aromatic information. Articles and resources at STKE highlight the evolution of olfactory signal transduction, describe the biosynthetic and signaling pathways of the plant hormone jasmonate, and provide an animated overview of how the plant hormone auxin regulates gene expression. PMID- 16478939 TI - How sensitive is a nose? AB - Odor sensitivity may not be due to odor-receptor (OR) binding but rather may be due to emergent properties of transduction pathways and the anatomical convergence of olfactory neurons. A recent study suggests that odor-OR interactions are brief and infrequently activate heterotrimeric GTP-binding proteins (G proteins); in contrast, visual receptors have long-lasting activation states and activate many G proteins. These differences may reflect strategies that evolved to accommodate very different signals, and the mechanisms described might be applicable for receptors across phyla. However, whereas visual receptors (rhodopsin) appeared before protostome-deuterostome separation, ORs may be independently derived in different phyla. Alternatively, phylum-distinct ORs may share common ancestry but be influenced by diversifying selection. Phylum distinct ORs may imply phylum-specific OR mechanisms, whereas common ancestry may imply common mechanisms. Nonetheless, most animals detect a similar repertoire of olfactory signals, and OR mechanisms may be convergent on those signals independent of receptor relatedness. Thus, recent insights into the molecular characteristics of odor perception in frogs may well be relevant to such processes as how mosquitoes detect host odors for a malaria-transmitting blood meal. PMID- 16478940 TI - Teaching resources. Model of the TIR1 pathway for auxin-mediated gene expression. AB - Auxin mediates numerous plant responses, some of which have been shown to require transcriptional regulation. One auxin response pathway, which depends on the relief of transcriptional repression, is mediated by TIR1 (transport inhibitor response protein 1). TIR1 is an auxin receptor and also a subunit of an SCF-type ubiquitin ligase. In the presence of a low concentration of auxin in the nucleus, members of the Aux/IAA family of transcriptional repressors bind to ARF proteins and inhibit the transcription of specific auxin response genes. Increased nuclear concentrations of auxin promote auxin binding to TIR1, causing the Aux/IAA proteins to associate with TIR1 and leading to their degradation by a proteasome mediated pathway. This decreases the concentration of Aux/IAA proteins in the nucleus and thereby enables the expression of certain auxin response genes. PMID- 16478942 TI - Cloning of pig parotid secretory protein gene upstream promoter and the establishment of a transgenic mouse model expressing bacterial phytase for agricultural phosphorus pollution control. AB - This study examined the feasibility of using the promoter of the pig parotid secretory protein (PSP) gene for expression of the phytase transgene in mouse models. The pig parotid secretory protein gene is specifically expressed at high levels in the salivary glands. The 10-kb upstream promoter region of the gene necessary for tissue-specific expression has been identified. We have constructed phytase transgenes composed of the appA phytase gene from Escherichia coli driven by the upstream promoter region of the pig PSP gene with a 3' tail of either bovine growth hormone or the pig PSP gene polyadenylation signal. Transgenic mouse models with the construct showed that the upstream region of the pig PSP gene is sufficient for directing the expression of phytase transgenes in the saliva. Expression of salivary phytase reduced fecal phytate by 8.5 and 12.5% in 2 transgenic mouse lines, respectively. These results suggest that the expression of phytase in salivary glands of monogastric animals offers a promising biological approach to relieve the requirement for dietary phosphate supplements and to reduce phosphorus pollution from animal agriculture. PMID- 16478943 TI - Effects of calpastatin and micro-calpain markers in beef cattle on tenderness traits. AB - The objective of this study was to assess the association of single nucleotide polymorphisms (SNP) developed at the calpastatin (CAST) and mu-calpain (CAPN1) genes with meat tenderness and palatability traits in populations with diverse genetic backgrounds. Three populations were used in the study. One population consisted of Bos taurus that included crossbred animals derived from Hereford, Angus, Red Angus, Limousin, Charolais, Gelbvieh, and Simmental (GPE7; n = 539). Another population consisted of Bos taurus with Bos indicus influence, including crossbred animals from Hereford, Angus, Brangus, Beefmaster, Bonsmara, and Romosinuano (GPE8; n = 580). The third population was Bos indicus and consisted of purebred Brahman (STARS; n = 444). Traits evaluated were meat tenderness measured as Warner-Bratzler shear force (WBSF; kg) at 14 d postmortem, and traits evaluated by trained sensory panels that included tenderness score, juiciness, and flavor intensity. A SNP at the CAST gene had a significant (P < 0.003) effect on WBSF and tenderness score in the GPE7 and GPE8 populations. Animals inheriting the TT genotype at CAST had meat that was more tender than those inheriting the CC genotype. The marker at the CAPN1 gene was significant (P < 0.03) for tenderness score in GPE7 and GPE8. Animals inheriting the CC genotype at CAPN1 had meat that was more tender than those inheriting the TT genotype. Markers at the CAST and CAPN1 genes were associated with flavor intensity in the GPE8 population. Animals inheriting the CC genotype at CAST and the TT genotype at CAPN1 produced steaks with an intense flavor when compared with the other genotypes. An interaction between CAST and CAPN1 was detected (P < 0.05) for WBSF on GPE8. The statistical significance of the interaction is questionable because of the limited number of observations in some cells. Markers developed at the CAST and CAPN1 genes are suitable for use in identifying animals with the genetic potential to produce meat that is more tender. PMID- 16478944 TI - Effects of quantitative trait loci on chromosomes 1, 2, 4, and 7 on growth, carcass, and meat quality traits in backcross Meishan x Large White pigs. AB - The aim of this work was to estimate whether genetic dissection of QTL on chromosomes 1, 2, 4, and 7, detected in an F2 Meishan x Large White population, can be achieved with a recombinant back-cross progeny test approach. For this purpose, a first generation of backcross (BC1) was produced by using frozen semen of F1 Large White x Meishan boars with Large White females. Four BC1 boars were selected because of their heterozygosity for at least 1 of the 4 regions. The BC1 boars were crossed with Large White sows, and the resulting BC2 offspring were measured for several growth and body composition traits. Contrary to the F2 animals, BC2 animals were also measured for meat quality traits in adductor, gluteus superficialis (GS), longissimus dorsi, and biceps femoris (BF) muscles. Each BC1 boar was tested for a total of 39 traits and for the 4 regions with statistical interval mapping analyses. The QTL effects obtained in BC1 families showed some differences compared with those described in F1 families. However, we confirmed QTL effects for growth in the SW1301-SW2512 markers interval on chromosome 1 and also for body composition in the SW1828-SW2512 markers interval on chromosome 1, in the SW2443-SWR783 markers interval on chromosome 2, and in the SW1369-SW632 markers interval on chromosome 7. In addition, we detected new QTL for growth traits on chromosome 2 and for meat quality traits on chromosomes 1 and 2. Growth of animals from weaning to the end of the test was influenced by the IGF2 gene region on chromosome 2. Concerning meat quality, ultimate pH of adductor, longissimus dorsi, and BF were affected by the interval delimited by UMNP3000 and SW2512 markers on chromosome 1, and a* of GS, L* of BF, and water holding capacity of GS were affected by QTL located between marker loci SW2443 and SWR783 on chromosome 2. Recombinant progeny testing appeared to be a suitable strategy for the genetic dissection of the QTL investigated. PMID- 16478945 TI - Genetic parameters for various random regression models to describe total sperm cells per ejaculate over the reproductive lifetime of boars. AB - The objective of this study was to model the variances and covariances of total sperm cells per ejaculate (TSC) over the reproductive lifetime of AI boars. Data from boars (n = 834) selected for AI were provided by Smithfield Premium Genetics. The total numbers of records and animals were 19,629 and 1,736, respectively. Parameters were estimated for TSC by age of boar classification with a random regression model using the Simplex method and DxMRR procedures. The model included breed, collector, and year-season as fixed effects. Random effects were additive genetic, permanent environmental effect of boar, and residual. Observations were removed when the number of data at a given age of boar classification was < 10 records. Preliminary evaluations showed the best fit with fifth-order polynomials, indicating that the best model would have fifth-order fixed regression and fifth-order random regressions for animal and permanent environmental effects. Random regression models were fitted to evaluate all combinations of first- through seventh-order polynomial covariance functions. Goodness of fit for the models was tested using Akaike's Information Criterion and the Schwarz Criterion. The maximum log likelihood value was observed for sixth-, fifth-, and seventh-order polynomials for fixed, additive genetic, and permanent environmental effects, respectively. However, the best fit as determined by Akaike's Information Criterion and the Schwarz Criterion was by fitting sixth-, fourth-, and seventh-order polynomials; and fourth-, second-, and seventh-order polynomials for fixed, additive genetic, and permanent environmental effects, respectively. Heritability estimates for TSC ranged from 0.27 to 0.48 across age of boar classifications. In addition, heritability for TSC tended to increase with age of boar classification. PMID- 16478946 TI - Genetic correlations between live yearling bull and steer carcass traits adjusted to different slaughter end points. 1. Carcass lean percentage. AB - We studied genetic relationships between age-constant live yearling beef bull growth and ultrasound traits and steer carcass traits with dissected steer carcass lean percentage adjusted to slaughter age-, HCW-, fat depth-, and marbling score-constant end points. Three measures of steer carcass lean percentage were used. Blue Tag lean percentage (BTLean) was predicted from HCW, fat depth, and LM area measurements. Ruler lean percentage (RulerLean) was predicted from carcass fat depth and LM depth and width measurements. Dissected lean percentage (DissLean) was based on dissection of the 10-11-12th rib section. Both BTLean (h2 = 0.30 to 0.44) and DissLean (h2 = 0.34 to 0.39) were more heritable than RulerLean (h2 = 0.05 to 0.14) at all end points. Genetic correlations among DissLean and RulerLean (rg = 0.61 to 0.70), DissLean and BTLean (rg = 0.56 to 0.72), and BTLean and RulerLean (rg = 0.59 to 0.90) indicated that these traits were not genetically identical. Adjusting Diss-Lean to different end points changed the magnitude, but generally not the direction, of genetic correlations with indicator traits. Ultrasound scan-age-constant live yearling bull lean percentage estimates were heritable (h2 = 0.26 to 0.42) and genetically correlated with each other (rg = 0.68 to 0.99) but had greater correlations with DissLean at slaughter age (rg = 0.24 to 0.48) and HCW (rg = 0.16 to 0.40) end points than at fat depth (rg = -0.08 to 0.13) and marbling score (rg = 0.02 to 0.11) end points. Scan-age-constant yearling bull ultrasound fat depth also had stronger correlations with DissLean at slaughter age (rg = 0.34) and HCW (rg = -0.25) than at fat depth (rg = -0.02) and marbling score (rg = -0.03) end points. Yearling bull scan-age-constant ultrasound LM area was positively correlated with DissLean at all endpoints (rg = 0.11 to 0.23). Genetic correlations between yearling bull LM method 1 width (rg = 0.38 to 0.56) and method 2 depth (rg = -0.17 to -0.38) measurements with DissLean suggested that LM shape may be a valuable addition to genetic improvement programs for carcass lean percentage at slaughter age, HCW, and fat depth constant end points. At all end points, steer carcass fat depth (rg = -0.60 to -0.64) and LM area (rg = 0.48 to 0.59) had stronger associations with DissLean than did corresponding live yearling bull measurements. Improved methods that combine live ultrasound and carcass traits would be beneficial for evaluating carcass lean percentage at fat depth or marbling score end points. PMID- 16478947 TI - Genetic correlations between live yearling bull and steer carcass traits adjusted to different slaughter end points. 2. Carcass fat partitioning. AB - Partial carcass dissection data from 1,031 finished crossbred beef steers were used to calculate heritabilities and genetic correlations among subcutaneous, intermuscular, and body cavity fat percentage and marbling score adjusted to slaughter age-, HCW-, fat depth-, and marbling score-constant endpoints. Genetic correlations were also calculated among these fat partitions with live growth and ultrasound traits evaluated in yearling beef bulls (n = 2,172) and steer carcass measurements. Heritabilities of the different fat partitions ranged from 0.22 (marbling score-constant body cavity fat) to 0.46 (HCW-constant marbling score). Genetic correlations between subcutaneous fat and intermuscular fat (rg = 0.16 to 0.32) and between intermuscular fat and body cavity fat (rg = 0.38 to 0.50) were more highly associated than subcutaneous fat and body cavity fat (rg = -0.08 to 0.05), indicating that fat depots are not under identical genetic control. Adjusting fat depots to different end points affected the magnitude but usually not the sign of the genetic correlations. Bull postweaning gain was associated with intermuscular (-0.24 to -0.35), body cavity (-0.24 to -0.29), and marbling fat (-0.24 to -0.39) in steers. Bull hip height was associated with body cavity ( 0.20 to -0.29) and marbling fat (-0.20 to -0.47) in steers. Bull ultrasound fat depth was associated with subcutaneous (0.11 to 0.29), intermuscular (0.05 to 0.36), body cavity (0.27 to 0.49), and marbling fat (0.27 to 0.73) in steers. Bull ultrasound intramuscular fat percentage was associated with subcutaneous ( 0.22 to -0.44) and intermuscular fat (-0.06 to 0.31) in steers. Bull ultrasound LM area was associated with body cavity (-0.25 to -0.31) and marbling fat (-0.25 to -0.30) in steers. Ultrasound LM width measurements were negatively correlated with subcutaneous fat (rg = -0.09 to -0.18), intermuscular fat (rg = -0.53 to 0.61), body cavity fat (rg = -0.63 to -0.69), and marbling score (rg = -0.75 to 0.87) at slaughter age-, HCW-, and fat depth-constant endpoints; correlations were generally lower at a marbling score-constant end point (rg = 0.07 to -0.49). Ultrasound indicator traits measured in seedstock may be useful in altering fat partitioning in commercial beef carcasses. PMID- 16478949 TI - Characterization of mammary gland development in pregnant gilts. AB - The purpose of this study was to quantify mammary gland (MG) growth during pregnancy in gilts and to determine the effect of anatomical location on gland growth. Size, composition, and histomorphology of MG were determined during gestation in 29 primigravid gilts. Gilts were allotted randomly to 6 slaughter groups: d 45 (n = 6), 60 (n = 4), 75 (n = 5), 90 (n = 4), 102 (n = 5), and 112 (n = 5) of gestation. Mammary glands were obtained at slaughter, and skin and extraneous fat pad were removed to obtain parenchymal MG tissue. Mammary glands were further separated into individual MG, and their locations were recorded. Individual MG were weighed and bisected in an approximate midsagittal section to measure cross-sectional area. Mammary glands were ground individually and pooled according to anatomical region: the first and second pairs of MG = anterior MG; the third, fourth, and fifth pairs of MG = middle MG; the sixth, seventh, and eighth pairs of MG = posterior MG. Contents of DM, CP, ether extract, and crude ash were measured. Wet weight, DM, CP, and ash content of total and individual MG increased (P < 0.01) between d 45 and 112 of gestation. Cross-sectional area of individual MG increased (P < 0.01) as gestation progressed. Percentage of CP and ash increased (P < 0.01), whereas percentage of ether extract decreased (P < 0.01) as gestation progressed. This inverse relationship between percentages of CP and ether extract (r = -0.999; P < 0.0001) was consistent with the histological shift from primarily an adipose tissue in early gestation to one containing extensive lobuloalveolar tissue in late gestation. Wet weight of middle MG was greater (P < 0.05) than that of posterior MG at d 102 and 112 of gestation, and amount of CP in middle MG was greater (P < 0.05) than that in anterior and posterior MG at d 102 and 112 of gestation, indicating that middle MG grow faster than other MG during late gestation. Rates of wet weight gain and protein accretion were accelerated (P < 0.01) after d 74 and 75 of gestation, respectively, indicating the importance of MG growth during the last trimester of gestation. The increase in rate of protein accretion after d 75 indicates a greater protein requirement for MG growth during later gestation. PMID- 16478948 TI - Ontogeny of T lymphocytes and intestinal morphological characteristics in neonatal pigs at different ages in the postnatal period. AB - To evaluate morphological characteristics and development of the immune system at different ages in neonatal pigs, 4 piglets were euthanized at 7, 14, and 18 d of age for collection of blood, bile, and intestinal tissue for morphological measurements. Blood was collected for differential cell counts, lymphocyte blastogenesis, immunoglobulin (Ig) concentrations, cytokine concentrations, and flow cytometric analysis. Bile was collected for quantification of Ig-A and Ig-M. Villus width and crypt depth from duodenum sections, as well as ileum crypt depth, were reduced (P < or = 0.08) in 18-d-old pigs compared with 7-d-old pigs. No age-related differences (P > or = 0.11) were observed in the number of goblet cells with neutral and acidic mucins, serum or enteric Ig concentrations, IL-2, IL-4, spontaneous lymphocyte proliferation, or leukocyte concentrations. When measured as counts per minute (cpm) and as a stimulation index (SI), lymphocyte proliferation responses to phytohaemagglutinin increased (P = 0.05) between 7 and 14 d of age; no changes (P = 0.10) occurred at 18 d of age. No age-related changes (P = 0.39) were observed in response to pokeweed mitogen (PWM) when measured as cpm; however, the SI from PWM-induced lymphocytes decreased (P = 0.04) 4-fold between 7 and 18 d of age. The CD4+:CD8+ and populations of lymphocytes expressing CD2+CD4+CD8- (T helper cells) and CD25+CD4+CD8- (activated T helper cells) were greater (P > or = 0.04) at 7 d of age than at 14 and 18 d. Populations of T lymphocytes, cytotoxic T cells (CD2+CD4-CD8+), activated lymphocytes (CD25+), and activated cytotoxic T cells (CD25+CD4-CD8+) were greater (P > or = 0.02) in 18-d-old pigs compared with 7-d-old pigs, whereas CD2+CD4-CD8- [double negative cells] were lower (P = 0.08) in 18-d-old pigs compared with 14-d old pigs. The percentage of CD2+ T cells was 8.4% at 7 d of age, and by the time the pigs reached 18 d of age, the percentage of CD2+ T cells was 33.8%. Moreover, the percentage of gammadelta T cells was greater (P = 0.02) in 18-d-old pigs than in 7-d-old pigs (74.8 vs. 46.1%, respectively). Results indicate that the porcine immune system and gut are continuously changing as the young pig matures. Changes occurred in lymphocyte phenotypic expression and functional capabilities, as well as morphology and mucin production, and their role may be to further protect the neonate from antigenic challenge as protection from passive immunity declines. PMID- 16478950 TI - Impacts of chronic stress and social status on various physiological and performance measures in pigs of different breeds. AB - Pigs typically experience various environmental stressors, which can negatively affect performance. Cortisol concentrations and various immune and performance measures are influenced by breed, but few data exist describing the impact of breed on stress responsiveness in pigs. The objective of this experiment was to determine if certain physiological responses to chronic stressors differed among 3 breeds and 2 commercial lines of pigs. The pigs were Landrace (n = 36), Meishan (n = 30), Yorkshire (n = 32), or 1 of 2 commercial lines (Line-A and Line-B; both n = 36). All pigs were weaned at 17 to 21 d and kept in a common nursery. At 49 d of age, pigs were assigned to 1 of 2 treatments: stress (heat, crowding, and mixing) or control (no stress treatment). Pigs were allocated to groups of 3 pigs per pen of the same sex. Control pigs were kept with their littermates. At the onset of the experiment, stressed pigs were mixed with 2 unfamiliar pigs once, and heat and crowding stressors were implemented simultaneously for 14 d. Pigs allocated to the stress treatment were video-recorded for 24 h following initiation of mixing to determine social status: dominant, intermediate, or submissive. Blood samples were taken at d 0 (baseline), 1, 7, and 14 to assess cortisol concentrations and immune measures. Breed and treatment affected cortisol, immune, and performance measures, but no significant breed x treatment interactions were found. In general, pigs subjected to the chronic stressor had lower (P < 0.001) BW and ADG (P < 0.001) than did control pigs. Plasma cortisol was lower (P < 0.001) among stressed pigs at d 7 and 14. Regardless of breed, lipopolysaccharide-induced proliferation (P < 0.01) and natural killer (NK; P < 0.005) cytotoxicity were greater in stressed pigs compared with controls. Furthermore, among stressed pigs, dominant pigs had a greater total white blood cell count (P < 0.005), NK (P < 0.05), and phagocytosis (P < 0.05) than the subordinate pigs. The results indicate that pig breed did not influence the physiological responses to the chronic concurrent stressors imposed for 14 d in this study, but social status did influence the immune responsiveness of these pigs to heat, crowding, and mixing. PMID- 16478941 TI - A global assembly of cotton ESTs. AB - Approximately 185,000 Gossypium EST sequences comprising >94,800,000 nucleotides were amassed from 30 cDNA libraries constructed from a variety of tissues and organs under a range of conditions, including drought stress and pathogen challenges. These libraries were derived from allopolyploid cotton (Gossypium hirsutum; A(T) and D(T) genomes) as well as its two diploid progenitors, Gossypium arboreum (A genome) and Gossypium raimondii (D genome). ESTs were assembled using the Program for Assembling and Viewing ESTs (PAVE), resulting in 22,030 contigs and 29,077 singletons (51,107 unigenes). Further comparisons among the singletons and contigs led to recognition of 33,665 exemplar sequences that represent a nonredundant set of putative Gossypium genes containing partial or full-length coding regions and usually one or two UTRs. The assembly, along with their UniProt BLASTX hits, GO annotation, and Pfam analysis results, are freely accessible as a public resource for cotton genomics. Because ESTs from diploid and allotetraploid Gossypium were combined in a single assembly, we were in many cases able to bioinformatically distinguish duplicated genes in allotetraploid cotton and assign them to either the A or D genome. The assembly and associated information provide a framework for future investigation of cotton functional and evolutionary genomics. PMID- 16478951 TI - The digestive fate of Escherichia coli glutamate dehydrogenase deoxyribonucleic acid from transgenic corn in diets fed to weanling pigs. AB - Corn containing genetically engineered plasmid DNA encoding an Escherichia coli glutamate dehydrogenase (gdhA) was fed to 19-d-old weanling swine to trace the digestive fate of the transgenic DNA. Eight pens of 8 pigs were fed a commercial (nongdhA) starter for 2 wk. One pig was randomly selected from each pen for 0-h control samples. The remaining 56 pigs were transitioned onto a corn-soybean meal diet and fed a diet containing 58% gdhA corn for approximately 1 wk; immediately thereafter, liver, 10th rib muscle, white blood cells, and plasma from the hepatic portal vein and ingesta from the stomach, distal ileum, and large intestine were collected. The DNA was extracted and the concentration determined via spectrophotometry. Polymerase chain reaction and gel electrophoresis were performed with primers designed to amplify 490 bp that included the plasmid's ligation site between the maize ubiquitin and the gdhA genes. The gdhA corn derived DNA and diet served as positive assay controls, and conventional corn DNA and distilled water acted as negative assay controls. Detection limits were 0.99 fg of target DNA confounded with 500 ng of conventional corn DNA per each 20 &L reaction. Transgenic DNA was detected in 71.43% of the stomach and 1.79% of the ileal ingesta samples from treatment animals but was not detected in the large intestine, white blood cells, plasma, liver, or muscle samples. Transgenic DNA was not detected in any sample from 0-h control animals. Stomach and ileal ingesta samples were further analyzed using real-time PCR. With an estimated limit of detection of 1.049 ag/microL, 89.29% of the stomach ingesta samples were positive (average 1.56 fg target DNA). The proportion of transgenic DNA to total DNA differed between diet and stomach ingesta samples (P < 0.001). Despite the greater sensitivity of real-time PCR, target DNA was detected in only 1.79% of ileal ingesta. These data suggest that the gdhA transgene began degradation in the stomach and was nondetectable in the large intestine. PMID- 16478952 TI - Performance, diarrhea incidence, and occurrence of Escherichia coli virulence genes during long-term administration of a probiotic Enterococcus faecium strain to sows and piglets. AB - As part of an interdisciplinary research project, the performance response of sows and their litters to the probiotic strain Enterococcus faecium NCIMB 10415, as well as some health characteristics of the piglets, were studied. Gestating sows (n = 26) were randomly allotted into 2 groups. The probiotic was administered by dietary supplementation to 1 group of sows and their respective litters (probiotic group), whereas the second group (control group) received no probiotic supplementation. The duration of the treatment was nearly 17 wk for sows (d 90 ante partum until d 28 postpartum) and 6 wk for piglets (d 15 to 56). Body weight and feed consumption were recorded weekly. The frequency of 4 toxin and 5 adhesion genes of putative pathogenic Escherichia coli was monitored weekly (d 7 to 35) by multiplex PCR assays, and fecal consistency of weaned piglets was studied daily. Probiotic treatment of lactating sows led to an overall pre weaning mortality of 16.2% compared with 22.3% in the control group (P = 0.44). Animal losses during the first 3 d of the suckling period were decreased in the probiotic group (P = 0.09). For piglets (n = 153), which were weaned at 28 d, there were no overall treatment differences in BW gain, feed intake, or feed efficiency. Probiotic supplementation, however, led to nearly a 40% reduction (P = 0.012). The actual percentage of piglets with postweaning diarrhea in the probiotic group was 21% compared with 38% in the control group (P = 0.05). The study on virulence factors of dominant fecal E. coli isolates revealed a high diversity with varying frequency and distribution of each single pathogenicity gene. The 440 isolates carried 29 different pathogenicity gene combinations as well as each of the 9 pathogenicity genes alone. Altogether, isolates with more than 2 pathogenicity genes were quite rare (< or = 10%), and up until d 28 isolates without any pathogenicity gene occurred most frequently. Depending on the time of sampling, one-third or more of all isolates contained est2 or est1b as single gene or in combination with other pathogenicity genes. PMID- 16478953 TI - Estimation of endogenous phosphorus loss in growing and finishing pigs fed semi purified diets. AB - Thirty-six barrows were used in a series of 3 P-balance experiments in which growing and finishing pigs were fed highly digestible, semi-purified diets at or below the dietary available P requirement to estimate the effect of BW on endogenous P loss. Experiments 1, 2, and 3 were conducted with pigs averaging 27, 59, and 98 kg of BW, respectively. In each experiment, pigs were placed in metabolism crates and allotted by weight and litter to 3 dietary treatments. The basal diet consisted of sucrose, dextrose, cornstarch, and casein fortified with minerals (except P) and vitamins. Diets 1, 2, and 3 in Exp. 1 were the basal diet with 0, 0.078, or 0.157% added P, respectively, from monosodium phosphate. In Exp. 2 and 3, diets 1, 2, and 3 were the basal diet with 0, 0.067, and 0.134% added P, respectively, from monosodium phosphate. Within replicate, pigs were fed equal amounts of feed twice daily. Pigs were adjusted to treatments for 7 d before a 6-d, marker-to-marker collection of feces and urine. Phosphorus intakes for pigs fed the 3 diets ranged from 1.73 to 3.91 g/d in Exp. 1, from 2.18 to 5.32 g/d in Exp. 2, and from 1.96 to 6.26 g/d in Exp. 3. Fecal P excretion and P absorption increased linearly (P < 0.05) with increasing P intake. In the 3 experiments, urinary P excretion (g/d) was low for pigs fed diet 1 (0.010, 0.011, 0.019) and diet 2 (0.013, 0.058, 0.084) and was low for pigs fed diet 3 in Exp. 1 (0.037); however, urinary P was greater in pigs fed diet 3 in Exp. 2 and 3 (0.550 and 0.486, respectively). When P absorption (Y, g/d) was regressed on P intake (X, g/d) in Exp. 1, 2, and 3, the relationships were linear (P < 0.01): Y = 0.110 + 0.971X (R2 = 0.999), Y = -0.156 + 0.939X (R2 = 0.998), and Y = -0.226 + 0.8919X (R2 = 0.982), respectively. Thus, our estimates of endogenous P loss at zero P intake were 110, 156, and 226 mg/d for 27-, 59-, and 98-kg pigs, respectively. When these Y-intercepts were regressed on BW, the relationship was Y = 63.06 + 1.632X (R2 = 0.996), where Y = endogenous P loss in mg/d and X = BW in kg. Based on these data, we estimate the endogenous P loss of pigs fed highly digestible, semi-purified diets to increase by approximately 1.632 mg for each 1 kg increase in BW from 25 to 100 kg. PMID- 16478955 TI - Influence of cobalt concentration on vitamin B12 production and fermentation of mixed ruminal microorganisms grown in continuous culture flow-through fermentors. AB - An experiment was conducted to determine the effects of dietary concentrations of Co on vitamin B12 production and fermentation of mixed ruminal microbes grown in continuous culture fermentors. Four fermentors were fed 14 g of DM/d. The DM consisted of a corn and cottonseed hull-based diet with Co supplemented as CoCO3. Dietary treatments were 1) control (containing 0.05 mg of Co/kg of DM), 2) 0.05 mg of supplemental Co/kg of DM, 3) 0.10 mg of supplemental Co/kg of DM, and 4) 1.0 mg of supplemental Co/kg of DM. After a 3-d adjustment period, fermentors were sampled over a 3-d sampling period. This process was repeated 2 additional times for a total of 3 runs. Ruminal fluid vitamin B12 concentrations were affected by Co supplementation (P < 0.01), and there was a treatment x day interaction (P < 0.01). By sampling d 3, cultures fed the basal diet supplemented with 0.10 mg of Co/kg had greater (P < 0.05) vitamin B12 concentrations than those supplemented with 0.05 mg of Co/kg of DM, and increasing supplemental Co from 0.10 to 1.0 mg/kg of DM increased (P < 0.01) ruminal fluid vitamin B12 concentration. Ruminal fluid succinate also was affected (P < 0.10) by a treatment x day interaction. Cobalt supplementation to the control diet greatly decreased (P < 0.05) succinate in ruminal cultures on sampling d 3 but not on d 1 or 2. Molar proportions of acetate, propionate, and isobutyrate, and acetate:propionate were not affected by the addition of supplemental Co to the basal diet. However, molar proportions of butyrate, valerate, and isovalerate increased (P < 0.05) in response to supplemental Co. The majority of long-chain fatty acids observed in this study were not affected by Co supplementation. However, percentages of C18:0 fatty acids in ruminal cultures tended (P < 0.10) to be greater for Co-supplemented diets relative to the control. Methane, ammonia, and pH were not greatly affected by Co supplementation. The results indicate that a total (diet plus supplemental) Co concentration of 0.10 to 0.15 mg/kg of dietary DM resulted in adequate vitamin B12 production to meet the requirements of ruminal microorganisms fed a high-concentrate diet in continuous flow fermentors. PMID- 16478954 TI - Estimation of true phosphorus digestibility and endogenous phosphorus loss in growing pigs fed conventional and low-phytate soybean meals. AB - This study reevaluated the method of regressing of total P output against dietary P intake to simultaneously estimate true P digestibility and endogenous P loss in growing pigs fed either conventional or low-phytate soybean meal (SBM). Four isocaloric diets were formulated to contain increasing concentrations of each type of SBM (8 diets total), and therefore contained increasing concentrations of dietary P. Dietary P and Ca concentrations were deficient because they were supplied solely by SBM, and Ca:total P ratios were less than 1:1. Sixteen barrows (initial BW 17.7 +/- 1.8 kg) were surgically fitted with a simple T-cannula at the distal ileum, randomly assigned to metabolism crates, and fed the experimental diets in a replicated 8 x 8 Latin square design. Feed was provided at 90 g/kg of BW(0.75) and fed in 2 equally sized meals at 0800 and 2000, with diets containing Cr sesquioxide (3 g/kg) as an indigestible marker. As the P concentration increased from 0.9 to 3.9 g/kg of DM, the apparent prececal P digestibility increased for conventional SBM (P < 0.05), but no relationship was observed for low-phytate SBM. The output of total P [mg/(kg of BW(0.75).d)], either prececal or total tract, exhibited a linear relationship (P < 0.01) with increasing P intake. However, a quadratic response (P = 0.02) was also detected for total tract P output from pigs fed low-phytate SBM. True P digestibility was not different between prececal and total tract collection sites (P > 0.10), but was greater (P < 0.01) for low-phytate SBM (62.6%) compared with conventional SBM (44.5%). Endogenous P estimates were not different between the SBM varieties and averaged 4.83 mg/(kg of BW(0.75).d). However, endogenous P estimates were highly variable between individual animals and, therefore, were not significantly different from zero. In this study, estimates of endogenous P loss from pigs were relatively low compared with previously reported values, and evidence of nonlinearity in P output was observed. These results suggest that the difference in true P digestibility between conventional SBM and low-phytate SBM is influenced by dietary phytate content when growing pigs are fed P-deficient diets. PMID- 16478956 TI - Increased puroindoline levels slow ruminal digestion of wheat (Triticum aestivum L.) starch by cattle. AB - Starch is the primary nutrient in ruminant diets used to promote high levels of performance. The site of starch digestion alters the nature of digestive end products (VFA in the rumen vs. glucose in the small intestine) and the efficiency of use. Cereal grain endosperm texture plays a major role in the rate and extent of starch degradation in ruminants. Wheat grain texture is regulated by the starch surface protein complex friabilin that consists primarily of puroindoline (PIN) A and B. Soft kernel texture in wheat is a result of both PIN genes being in the wild type active form and bound to starch. The objective of this study was to investigate the effect of varying PIN content in wheat on the rate of starch digestion in the rumen of beef cattle. In Exp. 1, 6 transgenic soft pin a/b isolines created in a hard wheat background, and 2 hard wheat controls were milled to yield a wide range of mean particle sizes across all lines. Milled samples were incubated in situ for 3 h. Increased expression of both PINA and PINB decreased DM digestibility (DMD) by 29.2% (P < 0.05) and decreased starch digestibility by 30.8% (P < 0.05). Experiment 2 separated the effects of particle size and total PIN content on digestion by milling the hardest and softest lines such that the mean particle size was nearly identical. Increased PIN decreased DMD by 21.7% (P < 0.05) and starch digestibility by 19.9% (P < 0.05) across particle sizes smaller than whole kernel. Experiment 3 addressed the time course of PIN effects in the rumen by observing ground samples of the hardest and softest lines over a 12-h in situ period. Increased PIN decreased DMD by 10.4% (P < 0.05) and starch digestibility by 11.0% (P < 0.05) across all time points. Dry matter and starch digestibility results demonstrated that increased expression of PIN was associated with a decreased rate of ruminal digestion independent of particle size. Puroindolines seem to aid in the protection of starch molecules from microbial digestion in the rumen, potentially increasing the amount of starch entering the small intestine. PMID- 16478957 TI - Estimation of undegradable intake protein in forages using neutral detergent insoluble nitrogen at a single in situ incubation time point. AB - Two experiments were conducted to evaluate the use of neutral detergent insoluble nitrogen (NDIN) at a single in situ incubation time point to estimate the undegradable intake protein (UIP) in forages as well as to compare rates of NDIN degradation. Forage samples in Exp. 1 comprised diet samples collected from range and meadow pastures monthly from May through September. In Exp. 2, clipped samples of alfalfa, birdsfoot trefoil, kura clover, and smooth bromegrass, and diet samples of the mixed legume-grass and smooth bromegrass were evaluated. Forage samples were incubated in situ for their mean retention time (MRT) estimated from IVDMD plus a 10-h passage lag to yield the total MRT (TMRT). Samples were also incubated for 0 h, 10 h, 75% TMRT, and 96 h. Undegradable intake protein was measured at 75% TMRT and TMRT, and calculated using fractional rates of degradation and passage with a 10-h passage lag. Rates of ruminal NDIN degradation were calculated using the slope of the regression of the natural logarithm of the potentially degradable NDIN remaining (96-h undegradable fraction subtracted) against time. The estimated UIP values obtained using 75% TMRT were highly correlated with those obtained using fractional rates of degradation and passage plus accounting for a 10-h passage lag in Exp. 1 (R2 = 0.95) and Exp. 2 (R2 = 0.98). Rates of NDIN degradation of range and meadow samples in Exp. 1 were slower (P < 0.05) from 0 to 10 h in May and June compared with rates from 10 h to 75% TMRT, but rates of degradation were not different (P = 0.34 to 0.71) for the rest of the collection periods. Rates of degradation were not different from 0 to 10 h and 10 h to 75% TMRT in Exp. 2 for diet (P = 0.82) or clipped samples (P = 0.86). The UIP of the forages in these experiments was accurately estimated using NDIN at a single in situ incubation time point equivalent to 75% of the TMRT, and rates of protein degradation can be obtained at this time point when 0- and 96-h incubations are included. PMID- 16478959 TI - Ruminal in situ disappearance kinetics of nitrogen and neutral detergent insoluble nitrogen from common crabgrass forages sampled on seven dates in northern Arkansas. AB - Southern crabgrass (Digitaria ciliaris [Retz.] Koel.) is often an undesirable species in field and forage crops, but visual observations suggest that livestock prefer it to many other summer forages. The objectives of this study were to assess the nutritive value of crabgrass sampled weekly between July 11 and August 22, 2001 and then to determine ruminal in situ disappearance kinetics of N and neutral detergent insoluble N (NDIN) for these forages. A secondary objective was to compare these kinetic estimates for crabgrass with those of alfalfa (Medicago sativa L.), bermudagrass (Cynodon dactylon [L.] Pers.), and or-chardgrass (Dactylis glomerata L.) as control hays. All kinetic evaluations were conducted with 5 ruminally cannulated Gelbvieh x Angus x Brangus steers (383 +/- 22.7 kg). Concentrations of N for crabgrass decreased linearly (P < or = 0.002) across sampling dates for leaf, stem, and whole-plant tissues. Conversely, percentages of the total N pool within NDIN and ADIN fractions generally increased over sampling dates in mostly linear patterns. For crabgrass, the immediately soluble portion of the total N pool (fraction A; overall mean = 54.6% of N) was greater (P < 0.001) than for all control hays. Crabgrass exhibited a more rapid N disappearance rate (overall mean = 0.093/h; expressed as a proportion disappearing/h) than that of bermudagrass (0.046/h; P < 0.001), but the disappearance rate for alfalfa N (0.223/h) was considerably faster (P < 0.001) than for crabgrass. The effective ruminal disappearance of N was greater (P < 0.001) for crabgrass (overall mean = 85.4%) than for the alfalfa (83.3%), bermudagrass (72.3%), or orchardgrass (76.0%) control hays. For alfalfa, the ruminal disappearance rate of NDIN (0.150/h) was more rapid (P < 0.001) than for crabgrass (overall mean = 0.110/h); however, the disappearance rate for crabgrass was faster than that for bermudagrass (0.072/h; P < 0.001) or for orchardgrass (0.098/h; P = 0.010). Effective ruminal disappearance of NDIN was greater (P < 0.001) for crabgrass (overall mean = 72.0%) than for the bermudagrass (69.0%) or alfalfa hays (50.5%), but there was no difference (P = 0.865) between crabgrass and orchardgrass (72.1%). Although crabgrass forages exhibited concentrations of total N that were comparable with those of alfalfa and rates of ruminal N disappearance that were < 50% of those for the alfalfa hay control, improvements in N use efficiency relative to alfalfa are questionable because of the excessively large Fraction A for crabgrass. PMID- 16478958 TI - Tolerance of inorganic selenium by range-type ewes during gestation and lactation. AB - The objectives of this 72-wk study were to evaluate and compare the effects of 6 dietary levels of inorganic Se on serum, whole blood, wool, and tissue Se concentrations and to determine the maximum tolerable level of Se for mature ewes during lamb production. Forty-one, 4-yr-old, range-type ewes (57.4 +/- 5.7 kg) were used in a completely randomized design with 6 dietary treatments. Sodium selenite was added to a corn and soybean meal-based diet to provide 0.2 (control), 4, 8, 12, 16, or 20 mg of dietary Se/kg to ewes during lamb production. Serum Se and ewe BW were measured at 4-wk intervals; whole blood Se and wool Se were measured every 12 wk; and samples of brain, diaphragm, heart, hoof, kidney, liver, and psoas major were collected at the termination of the experiment. Dietary Se did not affect ewe BW during the study (P = 0.69), and there was no treatment x time interaction. Serum Se increased linearly as dietary Se level increased (P < 0.001) and responded cubically (P = 0.02) over time. Selenium in whole blood increased linearly (P < 0.001) as supplemental Se increased. Wool Se increased linearly (P < 0.001) as dietary Se increased, and the response over time was quadratic (P < 0.001). Brain, diaphragm, heart, and psoas major Se increased (P < 0.05) linearly as dietary Se increased, liver Se responded quadratically (P < 0.05), and hoof and kidney Se increased cubicically (P < 0.05) as supplemental Se increased. In general, serum, whole blood, and tissue Se concentrations of ewes receiving 12, 16, or 20 mg of dietary Se/kg were greater (P < 0.05) than those of controls and ewes receiving less dietary Se. Although they were elevated in ewes receiving increased dietary Se, at no time did serum, whole blood, or wool Se concentrations reach levels previously reported as toxic, nor were clinical signs of Se toxicosis observed. Histopathological evaluation of liver, kidney, diaphragm, heart, and psoas major did not reveal evidence of Se toxicosis in ewes at any dietary Se level. Ewes under our experimental conditions and during the stresses of production were able to tolerate up to 20 mg of dietary Se/kg as sodium selenite for 72 wk. These findings suggest that the maximum tolerable level of inorganic Se for sheep is much greater than 2 mg/kg as was suggested previously. Experiments of longer duration and utilizing greater dietary Se concentrations are necessary to clearly define the maximum tolerable level. PMID- 16478960 TI - Influence of soybean hull supplementation on rumen fermentation and digestibility in steers consuming freshly clipped, endophyte-infected tall fescue. AB - Six steers (288.6 +/- 2.1 kg of BW) fitted with rumen and duodenal cannulas were used in a crossover design to evaluate intake, rumen fermentation, and site of nutrient digestion of freshly clipped, endophyte-infected (E+) Kentucky 31 tall fescue with or without soybean hull (SH) supplementation at 0.60% of BW (OM basis). Steers were placed in metabolism units within an environmentally controlled room and provided with free-choice access to fresh forage, water, and a vitamin/mineral supplement. The spring growth of E+ tall fescue was harvested daily during the experiment. Supplement was fed at 0700 with approximately 65% of the estimated daily forage. To maintain a fresh forage supply, additional forage was stored in a cooler and fed at 1900. Periods were 21 d with 14 d of adaptation and 7 d of digesta sample collection. Chromic oxide was used as a marker of duodenal digesta flow. Duodenal samples were taken 4 times daily with times shifting by 1 h each day to represent all 24 h of a day. Treatments were considered significant at P < 0.05. Supplementation of SH decreased forage OM intake from 1.64 to 1.41% of BW but increased total OM intake from 1.64 to 2.01% of BW. Apparent percentages (53.1%) and quantities (2,786 g/d) of rumen OM disappearance were not affected by supplementation. Percentages of total tract OM disappearance were not different (70.8%). Percentages of apparent rumen NDF disappearance also were not different (65.6%). Percentages of N disappearance were not different. Supplementation of SH resulted in increased total N (34.1 g/d) and microbial N (17.1 g/d) flowing to the duodenum. Rumen pH (6.5) was not affected, and rumen ammonia concentrations exhibited a time x treatment interaction in which SH decreased ammonia for 12 h after supplementation. Total VFA concentrations (103.9 mM) were unaffected. Liquid dilution rate (12.7%/h) and rumen OM fill (4.3 kg) were not different between treatments. Supplementation of SH at a rate of 0.60% of BW (OM basis) to calves consuming fresh E+ tall fescue decreased forage consumption but resulted in greater total intake, greater flow of N to the duodenum, and increased total tract OM disappearance. PMID- 16478961 TI - Relationship among GeneSTAR marbling marker, intramuscular fat deposition, and expected progeny differences in early weaned Simmental steers. AB - Research has demonstrated that triiodothyronine and thyroxin are correlated with marbling (MARB) deposition in Wagyu cattle. Polymorphisms in the 5' region of the thyroglobulin gene have been associated with an improvement in overall fattening and could be used as a gene marker for MARB. The commercially available GeneSTAR MARB test measures the specific thyroglobulin gene polymorphism and identifies cattle as having 0, 1, or 2 copies of the allele; these are identified as 0-STAR, 1-STAR, or 2-STAR, respectfully. Early weaned Simmental steers (n = 192) of known genetics were individually fed over a repeated 4-yr trial period to determine the correlations between GeneSTAR MARB test [Genetic Solutions/Bovigen Pty. Ltd. (Australia) in conjunction with Frontier Beef Systems, LLC (Louisville, CO)] results and intramuscular fat (IMF) deposition. Yearling weight, MARB, percent retail cuts, and carcass weight EPD were calculated for each steer. Steers were weaned at 88.0 +/- 1.1 d, pen-fed a high-concentrate diet for 84.5 +/- 0.4 d before allotment, and subsequently individually fed a 90% concentrate diet composed primarily of cracked corn and corn silage for 249.7 +/- 0.7 d. Steers were slaughtered at 423.3 +/- 1.4 d. Deoxyribonucleic acid samples were used by Genetic Solutions/Bovigen (Australia) for GeneSTAR MARB analysis. Steers with allele types of 0-STAR (n = 47), 1-STAR (n = 95), and 2-STAR (n = 33) had no effect (P > 0.10) on MARB score, chemically determined IMF percentage, quality grade, or percent low Choice and better. There were no differences (P > 0.10) in performance or other carcass parameters among the allele types. GeneSTAR results were not associated with MARB (P > 0.10). Conversely, MARB EPD was correlated (P < 0.01) with MARB score (r = 0.44) and IMF percentage (r = 0.27). Thus, in this management system, MARB EPD is an accurate predictor of IMF deposition. These data suggest that the GeneSTAR MARB marker was not an efficacious predictor of IMF deposition in early weaned Simmental steers fed a high-energy diet. PMID- 16478963 TI - Eating and drinking activity of newly weaned piglets: effects of individual characteristics, social mixing, and addition of extra zinc to the feed. AB - In production systems, piglets usually fast for a period after weaning, thereby increasing the risk of diarrhea and a reduction in growth. The low level of eating may relate to insufficient drinking activity, as solid feed intake must be accompanied by water intake. Mixing of newly weaned piglets is a well-known stressor and a common procedure in pig production. The effect of mixing on the temporal development of eating and drinking activity in newly weaned piglets has not been elucidated. High concentrations of zinc (Zn) in the feed improve the health and performance of piglets after weaning, but the underlying mechanisms are still obscure. One possibility is that Zn affects eating and drinking behavior. The effects of mixing 4 littermates from each of 2 litters and adding zinc oxide (ZnO; 2,500 ppm of Zn) to the feed were studied in a 2 x 2 factorial experiment using 123 piglets weaned at 27 d of age. Individual eating and drinking times during the initial 48 h after weaning were analyzed on 2 levels of aggregation, day and hour. The piglets spent less time eating on the first day after weaning compared with the second day (20 +/- 5 vs. 98 +/- 10 min, respectively; P < 0.001), whereas they spent more time drinking on the first day compared with the second day (13 +/- 1 vs. 9 +/- 0.5 min, respectively; P < 0.001). Eating and drinking times were positively associated (P < 0.001). Females ate for longer than males (61 +/- 8 vs. 44 +/- 7 min/24 h, respectively, P = 0.002), whereas sex did not affect drinking time. Drinking time increased (P = 0.003) and eating time decreased (P = 0.001) with increasing preweaning growth rate and weaning weight. Neither mixing nor addition of ZnO affected the daily eating time. However, nonmixed piglets given 2,500 ppm of Zn as ZnO in the feed spent more time drinking per day (12 +/- 1 min) than did nonmixed piglets offered 100 ppm of Zn as ZnO (10 +/- 1 min; P = 0.002). Mixing also affected the hourly distribution of the drinking activity (P < 0.05). In conclusion, the drinking behavior of newly weaned piglets was more affected by the external factors, mixing and addition of ZnO to the feed, than the eating behavior. As eating and drinking are strongly associated, more focus should be paid to the water intake and the interplay between eating and drinking behavior in future studies aiming to reduce weaning problems. PMID- 16478962 TI - Effects of polyvinyl chloride overwrap film, high-oxygen modified atmosphere packaging, or ultra-low-oxygen modified atmosphere packaging on bone marrow discoloration in beef humerus, rib, thoracic vertebra, and scapula. AB - Meat retailers have reported bone marrow discoloration to be a problem, especially in modified atmosphere packages (MAP). Therefore, it is important to determine the prevalence and cause(s) of bone marrow discoloration in different beef bones and packaging systems. Thirty-six beef humeri, ribs, scapulas, and thoracic vertebrae from USDA Select and Choice carcasses were obtained from a commercial abattoir, cut into 2.54-cm-thick sections at 4 d postmortem, and packaged into 1 of 3 systems: 1) polyvinyl chloride film (PVC) overwrap; 2) high oxygen (80% O2, 20% CO2) MAP; and 3) ultra-low-oxygen (70% N2, 30% CO2) MAP. Instrumental reflectance and visual color scores were taken on d 0, 2, and 4, and on d 0 to 4 of display, respectively. Bone marrow was extracted from humeri, ribs, and thoracic vertebrae for analysis but not from scapulas. Ribs, scapulas, and thoracic vertebrae packaged in PVC and high-oxygen MAP developed undesirable gray or black discoloration. In ultra-low-oxygen MAP, mean visual color scores were acceptable throughout the entire display period. Discoloration (darkening) was more extensive for ribs, scapulas, and thoracic vertebrae than for humeri, especially for bones packaged in PVC and high-oxygen MAP. Humeri had lower (P < 0.05) a* values (larger positive a* values indicate a redder color) than the other bones. The a* values for ribs, scapulas, and thoracic vertebrae decreased (P < 0.05) over time. Chroma showed that bone marrow discolored during display, but graying was dramatically less for all bones packaged in ultra-low-oxygen MAP and for humeri in PVC and high-oxygen MAP. Humeri marrow had lower (P < 0.05) 2 thiobarbituric acid reactive substances (TBARS) than did ribs and thoracic vertebrae marrow. Ultra-low-oxygen MAP resulted in the least amount of change in TBARS from d 0 to 4, whereas thoracic vertebrae marrow had greater (P < 0.05) TBARS values at d 4 of display than at d 0 in PVC and high-oxygen MAP. Humeri marrow had dramatically less total Fe and hemoglobin than did that of ribs and thoracic vertebrae for all packaging systems. Myoglobin was undetectable in humeri marrow. The much larger amounts of Fe and hemoglobin in ribs and thoracic vertebrae likely contribute to marrow discoloration. Bone marrow discoloration was distinct in ribs, scapulas, and thoracic vertebrae packaged in PVC or high oxygen MAP. Bones packaged in ultra-low-oxygen MAP had minimal discoloration. PMID- 16478964 TI - Environmental factors influencing heat stress in feedlot cattle. AB - Data from 3 summer feedlot studies were utilized to determine the environmental factors that influence heat stress in cattle and also to determine wind speed (WSPD; m.s(-1)) and solar radiation (RAD; W.m(-2)) adjustments to the temperature humidity index (THI). Visual assessments of heat stress, based on panting scores (0 = no panting to 4 = severe panting), were collected from 1400 to 1700. Mean daily WSPD, black globe temperature at 1500, and minimums for nighttime WSPD, nighttime black globe THI, and daily relative humidity were found to have the greatest influence on panting score from 1400 to 1700 (R2 = 0.61). From hourly values for THI, WSPD, and RAD, panting score was determined to equal -7.563 + (0.121 x THI) - (0.241 x WSPD) + (0.00082 x RAD) (R2 = 0.49). Using the ratio of WSPD to THI and RAD to THI (- 1.992 and 0.0068 for WSPD and RAD, respectively), adjustments to the THI were derived for WSPD and RAD. On the basis of these ratios and the average hourly data for 1400 to 1700, the THI, adjusted for WSPD and RAD, equals [4.51 + THI - (1.992 x WSPD) + (0.0068 x RAD)]. Four separate cattle studies, comparable in size, type of cattle, and number of observations to the 3 original studies, were utilized to evaluate the accuracy of the THI equation adjusted for WSPD and RAD, and the relationship between the adjusted THI and panting score. Mean panting score derived from individual observations of black-hided cattle in these 4 studies were 1.22, 0.94, 1.32, and 2.00 vs. the predicted panting scores of 1.15, 1.17, 1.30, and 1.96, respectively. Correlations between THI and panting score in these studies ranged from r = 0.47 to 0.87. Correlations between the adjusted THI and mean panting score ranged from r = 0.64 to 0.80. These adjustments would be most appropriate to use, within a day, to predict THI during the afternoon hours using hourly data or current conditions. In addition to afternoon conditions, nighttime conditions, including minimum WSPD, minimum black globe THI, and minimum THI, were also found to influence heat stress experienced by cattle. Although knowledge of THI alone is beneficial in determining the potential for heat stress, WSPD and RAD adjustments to the THI more accurately assess animal discomfort. PMID- 16478965 TI - Evaluation of the effects of dietary fat, conjugated linoleic acid, and ractopamine on growth performance, pork quality, and fatty acid profiles in genetically lean gilts. AB - An 8-wk study of the effects of CLA, rendered animal fats, and ractopamine, and their interactive effects on growth, fatty acid composition, and carcass quality of genetically lean pigs was conducted. Gilts (n = 228; initial BW of 59.1 kg) were assigned to a 2 x 2 x 3 factorial arrangement consisting of CLA, ractopamine, and fat treatments. The CLA treatment consisted of 1% CLA oil (CLA 60) or 1% soybean oil. Ractopamine levels were either 0 or 10 ppm. Fat treatments consisted of 0% added fat, 5% choice white grease (CWG), or 5% beef tallow (BT). The CLA and fat treatments were initiated at 59.1 kg of BW, 4 wk before the ractopamine treatments. The ractopamine treatments were imposed when the gilts reached a BW of 85.7 kg and lasted for the duration of the final 4 wk until carcass data were collected. Lipids from the belly, outer and inner layers of backfat, and LM were extracted and analyzed for fatty acid composition from 6 pigs per treatment at wk 4 and 8. Feeding CLA increased (P < 0.02) G:F during the final 4 wk. Pigs fed added fat as either CWG or BT exhibited decreased (P < 0.05) ADFI and increased (P < 0.01) G:F. Adding ractopamine to the diet increased (P < 0.01) ADG, G:F, and final BW. The predicted carcass lean percentage was increased (P < 0.05) in pigs fed CLA or ractopamine. Feeding either 5% fat or ractopamine increased (P < 0.05) carcass weight. Adding fat to the diets increased (P < 0.05) the 10th rib backfat depth but did not affect predicted percent lean. Bellies of gilts fed CLA were subjectively and objectively firmer (P < 0.01). Dietary CLA increased (P < 0.01) the concentration of saturated fatty acids and decreased (P < 0.01) the concentration of unsaturated fatty acids of the belly fat, both layers of backfat, and LM. Ractopamine decreased (P < 0.01) the i.m. fat content of the LM but had relatively little effect on the fatty acid profiles of the tissues compared with CLA. These results indicate that CLA, added fat, and ractopamine work mainly in an additive fashion to enhance pig growth and carcass quality. Furthermore, these results indicate that CLA results in more saturated fat throughout the carcass. PMID- 16478966 TI - Plasmid-mediated growth hormone-releasing hormone efficacy in reducing disease associated with Mycoplasma hyopneumoniae and porcine reproductive and respiratory syndrome virus infection. AB - The purpose of this study was to determine the effects of plasmid-mediated growth hormone releasing hormone (GHRH) supplementation on the clinical outcomes of pigs vaccinated against and challenged with either Mycoplasma hyopneumonia (M. hyo) and/or with porcine reproductive and respiratory syndrome (PRRS) virus. Before the first vaccination, pigs received a single i.m. injection of 0.625 mg of a porcine GHRH-expressing plasmid followed by electroporation of the injection site. Pigs were vaccinated at 2-wk intervals, challenged with either M. hyo and/or PRRS virus 2-wk after the second vaccination, and necropsied at 17 and 36 d after challenge. Clinical parameters associated with M. hyo challenge were improved with the GHRH treatment. Average daily gain between challenge and necropsy was improved (P = 0.04). Respiratory scores for M. hyo-challenged pigs tended to be lower in GHRH-treated animals compared to controls, and coughing scores were improved by the treatment (P = 0.01). Macroscopic lesions associated with M. hyo infection pneumonia were fewer in the group that received the GHRH expressing plasmid. No differences between treatment groups in the macroscopic pneumonia associated with PRRS virus were observed. No differences in serum antibodies to M. hyo or PRRS virus were observed with GHRH treatment. Nevertheless, IgG in the bronchioalveolar lavage was increased by the GHRH treatment in M. hyo-challenged animals (P < 0.03). The results of this study suggest that GHRH supplementation before vaccination may enhance the protection against M. hyo-induced pneumonia and that a single dose of GHRH-expressing plasmid was sufficient to elicit an improved clinical outcome in this disease challenge model. PMID- 16478967 TI - Effect of weaning age and commingling after the nursery phase of pigs in a wean to-finish facility on growth, and humoral and behavioral indicators of well being. AB - Pigs from one farrowing group in which gilts were bred to farrow pigs that would be either 14 or 21 d of age at weaning, were divided into older and younger age groups (108 pigs per group) and penned 12 pigs per pen in a wean-to-finish facility. At the end of the nursery phase, half the pigs in each age group were removed, rerandomized, and commingled for the finishing phase. The other half remained in their original pens. Pigs were fed common Phase 1 (d 0 to 14) and Phase 2 (d 14 to 35) nursery diets, and a common 4-phase program diet during growing/finishing, with transitions at 45, 68, and 90 kg of BW. The study ended when the lightest weight block averaged 107 kg. Blood was obtained on d 0, 2, 10, 27, 37, 44, and 65 after weaning to determine leukocyte concentrations. In addition, behavior was monitored during the nursery period at weaning (d 0), on d 7, 14, and 27 after weaning, and during the growing/finishing phase on d 35 (after commingling following the nursery phase), 38, 44, and 65 after weaning. Older pigs were heavier (P < 0.001) throughout the nursery period, and the BW difference between younger and older pigs increased from 2 to 6.5 kg at the start and end of the nursery period, respectively. Older pigs had a greater concentration of white blood cells (P < 0.05) and lymphocytes (P < 0.10) on d 0, 2, and 10 after weaning than younger pigs. Younger pigs spent less (P < 0.05) time resting on the day of weaning, and more (P < 0.05) time active during the overall nursery phase. During Phase 3 and in the overall finishing phase, younger pigs had greater (P < 0.01) ADG and G:F than older pigs. Moreover, during Phase 3, ADFI (as fed) decreased (P < 0.05) when older pigs were commingled compared with older pigs that were not commingled. There was no difference in ADFI of younger pigs, regardless of commingling (interaction; P < 0.10). Results of this study indicate that weaning age affects growth performance in a wean-to-finish facility, as well as behavioral and immunological responses to weaning and commingling after the nursery phase. Management strategies should be further explored to optimize these benefits without the detrimental effects on health observed during the nursery period in this study. PMID- 16478968 TI - Postnatal behavioral and physiological responses of piglets from gilts housed individually or in groups during gestation. AB - Gestational housing of sows remains a controversial issue that may affect the well-being of both sows and piglets. Therefore, 2 types of gestational housing were used to evaluate the stress imposed on pregnant gilts by each system and the effects on the offspring by comparing production, physiology, and behavioral measures of the piglets. Forty-eight Landrace x Yorkshire gilts were randomly assigned to groups (G) of 4 per pen (n = 8 pens; 3.9 m x 2.4 m) or to individual stalls (S; n = 16 stalls; 2.21 m x 0.61 m). Gilts were moved into individual farrowing crates 5 d before the expected farrowing date. Piglets were weighed at birth, d 14, and d 35. Two barrows from each litter were weaned at d 14 (early weaning) and housed together in pens. Maintenance behaviors (head in feeder, drinking, lying, eating mash) were videotaped and observed for the first 3 d after weaning using a 10-min interval scan sampling. Belly nosing and play/fight interactions were recorded from video observations for 3 d postweaning. An isolation test (30-min duration) was performed on one piglet from each pen of barrows on d 35. Time spent lying, the number of jumps against test box walls, and grunts and squeals were recorded in real time. Salivary cortisol was collected at 30-min intervals from baseline, and 0, 30, 60, and 90 min posttest. Jugular blood was collected from 2 barrows from each litter on d 1, 7, 14, 17, 21, and 28. Plasma TNF-alpha was analyzed by ELISA, and haptoglobin, alpha1-acid glycoprotein, and immunoglobulin G were analyzed by radial immunodiffusion. More piglets from the S treatment needed to be fed a liquid feed at weaning and drank more frequently on d 2 postweaning (P < 0.05). Additionally, by d 35 piglets from S gilts had a lighter BW (10.3 kg) than G piglets (12.8 kg; P < 0.01). Piglets from S gilts also grunted more during the 30-min isolation test (number of grunts = 356) than G piglets (number of grunts = 138; P < 0.01). Salivary cortisol and immune measures were not different. These data show some behavioral and production differences between piglets from individually stalled gilts and group housed gilts. Therefore, there may be production advantages to housing first parity gilts in groups. PMID- 16478969 TI - Identifying differences in feed efficiency among group-fed cattle. AB - Identification of efficient animals in the postweaning growth phase for use in selection for improved feed efficiency is important to improve the economic and environmental sustainability of the beef cattle industry. Progeny testing using group-fed animals in commercial feedlots is the most common and practical method used to evaluate postweaning growth on large numbers of animals. We developed the Cornell Value Discovery System (CVDS) to dynamically predict growth rate, accumulated weight, days required to reach target body composition, carcass weight, and composition of individual beef cattle fed in group pens. Observed BW, ADG, BW at 28% empty body fat (EBF), breed type, environmental conditions, and dietary ME concentration are used by the CVDS to predict, for each animal in a pen, the feed DM required for maintenance (FFM), the feed DM required for gain, and the total DM required for maintenance and gain (DMR). The CVDS then computes DMR-to-ADG ratio (DMR:ADG), which is a feed conversion measure, and ADG-to-DMR ratio (ADG:DMR), which is a feed efficiency measure, for each animal. This study used the observed F:G ratio of 362 individually fed steers to evaluate CVDS predicted indicators of feed efficiency and the Kleiber ratio. A subset of 37 data points was used to evaluate residual feed intake (RFI) as an indicator of feed efficiency. The database included 4 published studies, each with detailed individual animal description, environment, diet, and body composition information. The CVDS-predicted DMR:ADG accounted for 84% of the variation in the actual F:G ratio with a mean bias of 1.94% (P = 0.20). The predicted FFM to actual DMI ratio had a high correlation with actual ADG (R2 = 0.76), and indicated a decay-type nonlinear dilution of FFM as ADG increased. The CVDS predicted ADG:DMR and the Kleiber ratio had a significant (R2 = 0.88) logarithmic relationship. In an analysis of a contemporary group within the database, RFI was highly correlated with the F:G ratio (r = 0.71). There was a positive relationship between RFI and EBF. The RFIM (DMI - DMR) was moderately correlated with DMI and ADG (0.37 and -0.38; respectively), suggesting that selecting for low RFI(M) would decrease DMI and increase ADG in this database. We conclude that the CVDS model can be used to identify differences in the F:G and G:F ratios by predicting DMR for individual growing cattle fed in groups. PMID- 16478972 TI - Dynamics of mobilization and homing of endothelial progenitor cells after acute renal ischemia: modulation by ischemic preconditioning. AB - Endothelial progenitor cells (EPCs) have been shown to participate in tissue repair under diverse physiological and pathological conditions. It is unknown whether EPCs are mobilized in response to acute renal injury. The aim of this study was to characterize EPC mobilization and homing in the course of acute renal ischemia. Mice were subjected to unilateral renal artery clamping (UC) for 25 min. At 10 min, 3, 6, 24 h, and 7 days after UC, the pool of circulating and splenic CD34+/Flk-1+ cells within the monocytic population was detected by flow cytometry. For ischemic preconditioning (IPC), the first UC was performed 7 days before the repeated ischemic episode. For EPC detection in the kidney, cryosections were stained for c-Kit+/Tie-2+ cells. The number of circulating EPCs was not significantly affected at any time after UC compared with sham-operated or control mice. IPC did not significantly change the circulating pool of EPCs. Splenectomy performed before UC resulted in a surge of circulating EPCs. Accordingly, splenic EPCs were significantly increased after UC at 3 and 6 h, but not at later times. EPC homing to the spleen was absent in IPC animals. Immunohistochemical analysis of the kidneys showed a sixfold increase in the number of c-Kit+/Tie-2+ cells localized in the medullopapillary region in mice by day 7 after ischemia. Enriched population of c-Kit+/Tie-2+ cells from the medullopapillary parenchyma of Tie-2green fluorescent protein chimeric mice subjected to IPC was isolated and transplanted to wild-type mice with acute renal ischemia. This procedure resulted in the improvement of renal function in recipients. In conclusion, 1) renal ischemia rapidly (within 3-6 h) mobilizes EPCs, which transiently home to the spleen, acting as a temporary reservoir of mobilized EPCs; 2) the late phase of IPC is associated with the mobilization of the splenic pool and accumulation of EPCs in the renal medullopapillary region; and 3) transplantation of EPC-enriched cells from the medullopapillary parenchyma afforded partial renoprotection after renal ischemia, suggesting the role of the recruited EPCs in the functional rescue. PMID- 16478971 TI - Transport of estrone sulfate by the novel organic anion transporter Oat6 (Slc22a20). AB - Recently, a novel Slc22 gene family member expressed in murine olfactory mucosa was identified and based on sequence homology proposed to be an organic anion transporter [Oat6 (Slc22a20); J. C. Monte, M. A. Nagle, S. A. Eraly, and S. K. Nigam. Biochem Biophys Res Commun 323: 429-436, 2004]. However, no functional data for Oat6 was reported. In the present study, we demonstrate that murine Oat6 mediates the inhibitable transport of estrone sulfate using both Xenopus oocyte expression assay and Chinese hamster ovary (CHO) cells stably transfected with mOat6 (CHO-mOat6). Uptake was virtually eliminated by probenecid and the anionic herbicide 2,4-dichlorophenoxyacetate. The organic anions ochratoxin A, salicylate, penicillin G, p-aminohippurate, and urate inhibited mOat6-mediated accumulation to varying degrees. Transport of estrone sulfate by mOat6 was demonstrated to be saturable, and K(m) estimates of 109.8 +/- 22.6 microM in oocytes and 44.8 +/- 7.3 microM in CHO-mOat6 cells were obtained. Inhibitory constants for 2,4-dichlorophenoxyacetate (15.7 +/- 2.0 microM), salicylate (49.0 +/- 4.4 microM), probenecid (8.3 +/- 2.5 microM), and penicillin G (1,450 +/- 480 microM) were also determined. Accumulation of estrone sulfate mediated by mOat6 was significantly trans-stimulated by glutarate, indicating that mOat6 functions as an organic anion/dicarboxylate exchanger. These data demonstrate for the first time that the novel murine gene Oat6 (Slc22a20) encodes a functional organic anion transporter and mOat6 is indeed the newest member of the OAT gene family. PMID- 16478973 TI - Regulation of the Na-K-ATPase beta(1)-subunit promoter by multiple prostaglandin responsive elements. AB - Renal prostaglandins modulate the activity of a number of the transport systems in the kidney, including the Na-K-ATPase. Not only do prostaglandins have acute affects on renal Na-K-ATPase, but in addition prostaglandins have chronic affects, which include regulation at the transcriptional level. Previously, we have presented evidence that one such prostaglandin, PGE(1), stimulates the transcription of the human Na-K-ATPase beta(1)-subunit gene in Madin-Darby canine kidney cells via cAMP- and Ca(2+)-mediated pathways (Taub M, Borsick M, Geisel J, Matlhagela K, Rajkhowa T, and Allen C. Exp Cell Res 299: 1-14, 2004; Matlhagela K, Borsick M, Rajkhowa T, and Taub M. J Biol Chem 280: 334-346, 2005). Evidence was presented indicating that PGE(1) stimulation was mediated through the binding of cAMP-regulatory element binding protein (CREB) to a prostaglandin-responsive element (PGRE) as well as Sp1 binding to an adjacent Sp1 site. In this report, we present evidence from EMSAs and DNA affinity precipitation studies that another PGRE present in the Na-K-ATPase beta(1)-subunit promoter similarly binds CREB and Sp1. The evidence that indicates a requirement for CREB as well as Sp1 for gene activation through both PGREs (PGRE1 and PGRE3) includes studies with a dominant negative CREB (KCREB), Drosophila SL2 cells, and PGRE mutants. The results of these studies are indicative of a synergism between Sp1 and CREB in mediating regulation by PGRE3; while regulation occurring through PGRE1 also involves Sp1 and CREB, the mechanism appears to be distinct. PMID- 16478974 TI - Effects of water restriction on gene expression in mouse renal medulla: identification of 3betaHSD4 as a collecting duct protein. AB - To identify novel gene targets of vasopressin regulation in the renal medulla, we performed a cDNA microarray study on the inner medullary tissue of mice following a 48-h water restriction protocol. In this study, 4,625 genes of the possible approximately 12,000 genes on the array were included in the analysis, and of these 157 transcripts were increased and 63 transcripts were decreased by 1.5 fold or more. Quantitative, real-time PCR measurements confirmed the increases seen for 12 selected transcripts, and the decreases were confirmed for 7 transcripts. In addition, we measured transcript abundance for many renal collecting duct proteins that were not represented on the array; aquaporin-2 (AQP2), AQP3, Pax-8, and alpha- and beta-Na-K-ATPase subunits were all significantly increased in abundance; the beta- and gamma-subunits of ENaC and the vasopressin type 1A receptor were significantly decreased. To correlate changes in mRNA expression with changes in protein expression, we carried out quantitative immunoblotting. For most of the genes examined, changes in mRNA abundances were not associated with concomitant protein abundance changes; however, AQP2 transcript abundance and protein abundance did correlate. Surprisingly, aldolase B transcript abundance was increased but protein abundance was decreased following 48 h of water restriction. Several transcripts identified by microarray were novel with respect to their expression in mouse renal medullary tissues. The steroid hormone enzyme 3beta-hydroxysteroid dehydrogenase 4 (3betaHSD4) was identified as a novel target of vasopressin regulation, and via dual labeling immunofluorescence we colocalized the expression of this protein to AQP2-expressing collecting ducts of the kidney. These studies have identified several transcripts whose abundances are regulated in mouse inner medulla in response to an increase in endogenous vasopressin levels and could play roles in the regulation of salt and water excretion. PMID- 16478975 TI - Anti-inflammatory and antinecrotic effects of the volatile anesthetic sevoflurane in kidney proximal tubule cells. AB - Renal ischemia-reperfusion (IR) injury is a major clinical problem without effective therapy. We recently reported that volatile anesthetics protect against renal IR injury, in part, via their anti-inflammatory properties. In this study, we demonstrate the anti-inflammatory and antinecrotic effects of sevoflurane in cultured kidney proximal tubule cells and probed the mechanisms of sevoflurane induced renal cellular protection. To mimic inflammation, human kidney proximal tubule (HK-2) cells were treated with tumor necrosis factor-alpha (TNF-alpha; 25 ng/ml) in the presence or absence of sevoflurane. In addition, we studied the effects of sevoflurane pretreatment on hydrogen peroxide (H2O2)-induced necrotic cell death in HK-2 or porcine proximal tubule (LLC-PK1) cells. We demonstrate that sevoflurane suppressed proinflammatory effects of TNF-alpha evidenced by attenuated upregulation of proinflammatory cytokine mRNA (TNF-alpha, MCP-1) and ICAM-1 protein and reduced nuclear translocation of the proinflammatory transcription factors NF-kappaB and AP-1. Sevoflurane reduced necrotic cell death induced with H2O2 in HK-2 cells as well as in LLC-PK1 cells. Sevoflurane treatment resulted in phosphorylation of prosurvival kinases, ERK and Akt, and increased de novo HSP-70 protein synthesis without affecting the synthesis of HSP 27 or HSP-32. We conclude that sevoflurane has direct anti-inflammatory and antinecrotic effects in vitro in a renal cell type particularly sensitive to injury following IR injury. These mechanisms may, in part, account for volatile anesthetics' protective effects against renal IR injury. PMID- 16478976 TI - AT1 receptor-mediated accumulation of extracellular angiotensin II in proximal tubule cells: role of cytoskeleton microtubules and tyrosine phosphatases. AB - Long-term angiotensin II (ANG II) administration is associated with increased ANG II accumulation in the kidney, but intrarenal compartment(s) involved in this response remains to be determined. We tested the hypothesis that 1) extracellular ANG II is taken up by proximal tubule cells (PTCs) through AT(1) receptor mediated endocytosis, 2) this process is regulated by cytoskeleton microtubule- and tyrosine phosphatase-dependent mechanisms, and 3) AT(1) receptor-mediated endocytosis of ANG II has a functional relevance by modulating intracellular cAMP signaling. In cultured PTCs, [(125)I]Tyr-labeled ANG II and fluorescein labeled ANG II were internalized in a time-dependent manner and colocalized with the endosome marker Alexa Fluor 594-transferrin. Endocytosis of extracellular ANG II was inhibited by the AT(1) receptor blocker losartan (16.5 +/- 4.6%, P < 0.01 vs. ANG II, 78.3 +/- 6.2%) and by the tyrosine phosphatase inhibitor phenylarsine oxide (PAO; 30.0 +/- 3.5%, P < 0.05 vs. ANG II). Intracellular ANG II levels were increased by approximately 58% (basal, 229.8 +/- 11.4 vs. ANG II, 361.3 +/- 11.8 pg ANG II/mg protein, P < 0.01), and the responses were blocked by losartan (P < 0.01), the cytoskeleton microtubule inhibitor colchicine (P < 0.05), and PAO (P < 0.01), whereas depletion of clathrin-coated pits with hyperosmotic sucrose had no effect (356.1 +/- 25.5 pg ANG II/mg protein, not significant). ANG II accumulation was associated with significant inhibition of both basal (control, 15.5 +/- 2.8 vs. ANG II, 9.1 +/- 2.4 pmol/mg protein, P < 0.05) and forskolin stimulated cAMP signaling (forskolin, 68.7 +/- 8.6 vs. forskolin + ANG II, 42.8 +/- 13.8 pmol/mg protein, P < 0.01). These effects were blocked by losartan and PAO. We conclude that extracellular ANG II is internalized in PTCs through AT(1) receptor-mediated endocytosis and that internalized ANG II may play a functional role in proximal tubule cells by inhibiting intracellular cAMP signaling. PMID- 16478977 TI - Tempol reduces oxidative stress and restores renal dopamine D1-like receptor- G protein coupling and function in hyperglycemic rats. AB - Dopamine via activation of renal D1-like receptors inhibits the activities of Na K-ATPase and Na/H exchanger and subsequently increases sodium excretion. Decreased renal dopamine production and sodium excretion are associated with hyperglycemic conditions. We have earlier reported D1-like receptor-G protein uncoupling and reduced response to D1-like receptor activation in streptozotocin (STZ)-treated hyperglycemic rats (Marwaha A, Banday AA, and Lokhandwala MF. Am J Physiol Renal Physiol 286: F451-F457, 2004). The present study was designed to test the hypothesis that oxidative stress associated with hyperglycemia increases basal D1-like receptor serine phosphorylation via activation of the PKC-G protein receptor kinase (GRK) pathway, resulting in loss of D1-like receptor-G protein coupling and function. We observed that STZ-treated rats exhibited oxidative stress as evidenced by increased lipid peroxidation. Furthermore, PKC activity and expression of PKC-betaI- and -delta-isoforms were increased in STZ-treated rats. In addition, in STZ-treated rats there was increased GRK2 translocation to proximal tubular membrane and increased basal serine D1-like receptor phosphorylation. Supplementation with the antioxidant tempol lowered oxidative stress in STZ-treated rats, led to normalization of PKC activity, and prevented GRK2 translocation. Furthermore, tempol supplementation in STZ-treated rats restored D1-like receptor-G protein coupling and inhibition of Na-K-ATPase activity on D1-like receptor agonist stimulation. The functional consequence was the restoration of the natriuretic response to D1-like receptor activation. We conclude that oxidative stress associated with hyperglycemia causes an increase in activity and expression of PKC. This leads to translocation of GRK2, subsequent phosphorylation of the D1-like receptor, its uncoupling from G proteins and loss of responsiveness to agonist stimulation. PMID- 16478978 TI - Gamble's "economy of water" revisited: studies in urea transporter knockout mice. AB - The Gamble phenomenon (initially described over 70 years ago as "an economy of water in renal function referable to urea") suggested that urea plays a special role in the urinary concentrating mechanism and that the concentrating mechanism depends in some complex way on an interaction between NaCl and urea. In this study, the role of collecting duct urea transporters in the Gamble phenomenon was investigated in wild-type mice and mice in which the inner medulla collecting duct (IMCD) facilitative urea transporters, UT-A1 and UT-A3, had been deleted (UT A1/3-/- mice). The general features of the Gamble phenomenon were confirmed in wild-type mice, namely 1) the water requirement for the excretion of urea is less than for the excretion of an osmotically equivalent amount of NaCl; and 2) when fed various mixtures of urea and salt in the diet, less water is required for the excretion of the two substances together than the amount of water needed for the excretion of the two substances separately. In UT-A1/3-/- mice both of these elements of the phenomenon were absent, indicating that IMCD urea transporters play a central role in the Gamble phenomenon. A titration study in which wild type mice were given progressively increasing amounts of urea showed that the ability of the kidney to reabsorb urea was saturable, resulting in osmotic diuresis above excretion rates of approximately 6,000 microosmol/day. In the same titration experiments, when increasing amounts of NaCl were added to the diet, mice were unable to increase urinary NaCl concentrations to >420 mM, resulting in osmotic diuresis at NaCl excretion rates of approximately 3,500 microosmol/day. Thus both urea and NaCl can induce osmotic diuresis when large amounts are given, supporting the conclusion that the decrease in water excretion with mixtures of urea and NaCl added to the diet (compared with pure NaCl or urea) is due to the separate abilities of urea and NaCl to induce osmotic diuresis, rather than to any specific interaction of urea transport and NaCl transport at an epithelial level. PMID- 16478979 TI - Adenosine2A receptor vasodilation of rat preglomerular microvessels is mediated by EETs that activate the cAMP/PKA pathway. AB - Dilation of rat preglomerular microvessels (PGMV) by activation of adenosine A2A receptors (A2AR) is coupled to epoxyeicosatrienoic acid (EET) release. We have investigated the commonality of this signal transduction pathway, i.e., sequential inhibition of G(salpha), adenylyl cyclase, PKA, and Ca2+-activated K+ (KCa) channel activity, to the vasoactive responses to A2AR activation by a selective A2A agonist, CGS-21680, compared with those of 11,12-EET. Male Sprague Dawley rats were anesthetized, and microdissected arcuate arteries (110-130 microm) were cannulated and pressurized to 80 mmHg. Vessels were superfused with Krebs solution containing NG-nitro-L-arginine methyl ester (L-NAME) and indomethacin and preconstricted with phenylephrine. We assessed the effect of 3 aminobenzamide (10 microM), an inhibitor of mono-ADP-ribosyltranferases, on responses to 11,12-EET (3 nM) and CGS-21680 (10 microM) and found that both were inhibited by approximately 70% (P<0.05), whereas the response to SNP (10 microM) was unaffected. Furthermore, 11,12-EET (100 nM), like cholera toxin (100 ng/ml), stimulated ADP-ribose formation in homogenates of arcuate arteries compared with control. SQ-22536 (10 microM), an inhibitor of adenylyl cyclase activity, and myristolated PKI (14-22) amide (5 microM), an inhibitor of PKA, decreased activity of 11,12-EET and CGS-21680. Incubation of 11,12-EET (3 nM-3 microM) with PGMV resulted in an increase in cAMP levels (P<0.05). The responses to both 11,12 EET and CGS-21680 were significantly reduced by superfusion of iberiotoxin (100 nM), an inhibitor of KCa channel activity. Thus in rat PGMV activation of A2AR is coupled to EET release upstream of adenylyl cyclase activation and EETs stimulate mono-ADP-ribosyltransferase, resulting in Gsalpha protein activation. PMID- 16478980 TI - Unique role for the UbL-UbA protein Ddi1 in turnover of SCFUfo1 complexes. AB - SCF complexes are E3 ubiquitin-protein ligases that mediate degradation of regulatory and signaling proteins and control G1/S cell cycle progression by degradation of G1 cyclins and the cyclin-dependent kinase inhibitor, Sic1. Interchangeable F-box proteins bind the core SCF components; each recruits a specific subset of substrates for ubiquitylation. The F-box proteins themselves are rapidly turned over by autoubiquitylation, allowing rapid recycling of SCF complexes. Here we report a role for the UbL-UbA protein Ddi1 in the turnover of the F-box protein, Ufo1. Ufo1 is unique among F-box proteins in having a domain comprising multiple ubiquitin-interacting motifs (UIMs) that mediate its turnover. Deleting the UIMs leads to stabilization of Ufo1 and to cell cycle arrest at G1/S of cells with long buds resembling skp1 mutants. Cells accumulate substrates of other F-box proteins, indicating that the SCF pathway of substrate ubiquitylation is inhibited. Ufo1 interacts with Ddi1 via its UIMs, and Deltaddi1 cells arrest when full-length UFO1 is overexpressed. These results imply a role for the UIMs in turnover of SCF(Ufo1) complexes that is dependent on Ddi1, a novel activity for an UbL-UbA protein. PMID- 16478981 TI - A CTCF-dependent silencer located in the differentially methylated area may regulate expression of a housekeeping gene overlapping a tissue-specific gene domain. AB - The tissue-specific chicken alpha-globin gene domain represents one of the paradigms, in terms of its constitutively open chromatin conformation and the location of several regulatory elements within the neighboring housekeeping gene. Here, we show that an 0.2-kb DNA fragment located approximately 4 kb upstream to the chicken alpha-globin gene cluster contains a binding site for the multifunctional protein factor CTCF and possesses silencer activity which depends on CTCF binding, as demonstrated by site-directed mutagenesis of the CTCF recognition sequence. CTCF was found to be associated with this recognition site in erythroid cells but not in lymphoid cells where the site is methylated. A functional promoter directing the transcription of the apparently housekeeping ggPRX gene was found 120 bp from the CTCF-dependent silencer. The data are discussed in terms of the hypothesis that the CTCF-dependent silencer stabilizes the level of ggPRX gene transcription in erythroid cells where the promoter of this gene may be influenced by positive cis-regulatory signals activating alpha globin gene transcription. PMID- 16478982 TI - DNA damage during reoxygenation elicits a Chk2-dependent checkpoint response. AB - Due to the abnormal vasculature of solid tumors, tumor cell oxygenation can change rapidly with the opening and closing of blood vessels, leading to the activation of both hypoxic response pathways and oxidative stress pathways upon reoxygenation. Here, we report that ataxia telangiectasia mutated-dependent phosphorylation and activation of Chk2 occur in the absence of DNA damage during hypoxia and are maintained during reoxygenation in response to DNA damage. Our studies involving oxidative damage show that Chk2 is required for G2 arrest. Following exposure to both hypoxia and reoxygenation, Chk2-/- cells exhibit an attenuated G2 arrest, increased apoptosis, reduced clonogenic survival, and deficient phosphorylation of downstream targets. These studies indicate that the combination of hypoxia and reoxygenation results in a G2 checkpoint response that is dependent on the tumor suppressor Chk2 and that this checkpoint response is essential for tumor cell adaptation to changes that result from the cycling nature of hypoxia and reoxygenation found in solid tumors. PMID- 16478983 TI - A role for gcn5-mediated global histone acetylation in transcriptional regulation. AB - Transcriptional activators often require histone acetyltransferases (HATs) for full activity. The common explanation is that activators directly recruit HATs to gene promoters to locally hyperacetylate histones and thereby facilitate transcription complex formation. However, in addition to being targeted to specific loci, HATs such as Gcn5 also modify histones genome-wide. Here we provide evidence for a role of this global HAT activity in regulated transcription. We show that activation by direct recruitment of the transcriptional machinery neither recruits Gcn5 nor induces changes in histone acetylation yet can strongly depend on Gcn5 at promoters showing a high basal state of Gcn5-mediated histone acetylation. We also show that Gcn5 dependency varies among core promoters and is influenced by the strength of interaction used to recruit the machinery and by the affinity of the latter for the core promoter. These data support a role for global Gcn5 HAT activity in modulating transcription independently of its known coactivator function. PMID- 16478985 TI - The UL69 transactivator protein of human cytomegalovirus interacts with DEXD/H Box RNA helicase UAP56 to promote cytoplasmic accumulation of unspliced RNA. AB - The UL69 gene product of human cytomegalovirus belongs to a family of regulatory proteins conserved among all herpesviruses that have in part been characterized as posttranscriptional transactivators participating in the nuclear export of RNA. Recent experiments suggested that pUL69 also acts as a posttranscriptional activator since it was demonstrated that nucleocytoplasmic shuttling via a CRM1 independent nuclear export signal is a prerequisite for its stimulatory effect on gene expression. Based on these findings we initiated studies to investigate the role of pUL69 in mRNA export and demonstrate that pUL69 efficiently promotes the cytoplasmic accumulation of unspliced RNA. Furthermore, we show that this pUL69 activity is linked to the cellular mRNA export machinery by direct protein interaction with the highly related DEXD/H-box RNA helicases UAP56 and URH49. Particularly, we identified a 12-amino-acid domain within the N terminus of pUL69 which is required for binding to UAP56 and URH49, and we could demonstrate that UAP56 interaction and nucleocytoplasmic shuttling are both prerequisites for pUL69-mediated mRNA export. Thus, we identified a novel cellular target which provides a herpesviral regulatory protein with access to a conserved cellular transport system in order to promote nuclear export of unspliced RNA. PMID- 16478984 TI - The Nse5-Nse6 dimer mediates DNA repair roles of the Smc5-Smc6 complex. AB - Stabilization and processing of stalled replication forks is critical for cell survival and genomic integrity. We characterize a novel DNA repair heterodimer of Nse5 and Nse6, which are nonessential nuclear proteins critical for chromosome segregation in fission yeast. The Nse5/6 dimer facilitates DNA repair as part of the Smc5-Smc6 holocomplex (Smc5/6), the basic architecture of which we define. Nse5-Nse6 [corrected] (Nse5 and Nse6) [corrected] mutants display a high level of spontaneous DNA damage and mitotic catastrophe in the absence of the master checkpoint regulator Rad3 (hATR). Nse5/6 mutants are required for the response to genotoxic agents that block the progression of replication forks, acting in a pathway that allows the tolerance of irreparable UV lesions. Interestingly, the UV sensitivity of Nse5/6 [corrected] is suppressed by concomitant deletion of the homologous recombination repair factor, Rhp51 (Rad51). Further, the viability of Nse5/6 mutants depends on Mus81 and Rqh1, factors that resolve or prevent the formation of Holliday junctions. Consistently, the UV sensitivity of cells lacking Nse5/6 can be partially suppressed by overexpressing the bacterial resolvase RusA. We propose a role for Nse5/6 mutants in suppressing recombination that results in Holliday junction formation or in Holliday junction resolution. PMID- 16478986 TI - Gadd34 requirement for normal hemoglobin synthesis. AB - The protein encoded by growth arrest and DNA damage-inducible transcript 34 (Gadd34) is associated with translation initiation regulation following certain stress responses. Through interaction with the protein phosphatase 1 catalytic subunit (PP1c), Gadd34 recruits PP1c for the removal of an inhibitory phosphate group on the alpha subunit of elongation initiation factor 2, thereby reversing the shutoff of protein synthesis initiated by stress-inducible kinases. In the absence of stress, the physiologic consequences of Gadd34 function are not known. Initial analysis of Gadd34-null mice revealed several significant findings, including hypersplenism, decreased erythrocyte volume, increased numbers of circulating erythrocytes, and decreased hemoglobin content, resembling some thalassemia syndromes. Biochemical analysis of the hemoglobin-producing reticulocyte (an erythrocyte precursor) revealed that the decreased hemoglobin content in the Gadd34-null erythrocyte is due to the reduced initiation of the globin translation machinery. We propose that an equilibrium state exists between Gadd34/PP1c and the opposing heme-regulated inhibitor kinase during hemoglobin synthesis in the reticulocyte. PMID- 16478987 TI - Acetylation of human 8-oxoguanine-DNA glycosylase by p300 and its role in 8 oxoguanine repair in vivo. AB - The human 8-oxoguanine-DNA glycosylase 1 (OGG1) is the major DNA glycosylase responsible for repair of 7,8-dihydro-8-oxoguanine (8-oxoG) and ring-opened fapyguanine, critical mutagenic DNA lesions that are induced by reactive oxygen species. Here we show that OGG1 is acetylated by p300 in vivo predominantly at Lys338/Lys341. About 20% of OGG1 is present in acetylated form in HeLa cells. Acetylation significantly increases OGG1's activity in vitro in the presence of AP-endonuclease by reducing its affinity for the abasic (AP) site product. The enhanced rate of repair of 8-oxoG in the genome by wild-type OGG1 but not the K338R/K341R mutant, ectopically expressed in oxidatively stressed OGG1-null mouse embryonic fibroblasts, suggests that acetylation increases OGG1 activity in vivo. At the same time, acetylation of OGG1 was increased by about 2.5-fold after oxidative stress with no change at the polypeptide level. OGG1 interacts with class I histone deacetylases, which may be responsible for its deacetylation. Based on these results, we propose a novel regulatory function of OGG1 acetylation in repair of its substrates in oxidatively stressed cells. PMID- 16478988 TI - Abnormal expression of REST/NRSF and Myc in neural stem/progenitor cells causes cerebellar tumors by blocking neuronal differentiation. AB - Medulloblastoma, one of the most malignant brain tumors in children, is thought to arise from undifferentiated neural stem/progenitor cells (NSCs) present in the external granule layer of the cerebellum. However, the mechanism of tumorigenesis remains unknown for the majority of medulloblastomas. In this study, we found that many human medulloblastomas express significantly elevated levels of both myc oncogenes, regulators of neural progenitor proliferation, and REST/NRSF, a transcriptional repressor of neuronal differentiation genes. Previous studies have shown that neither c-Myc nor REST/NRSF alone could cause tumor formation. To determine whether c-Myc and REST/NRSF act together to cause medulloblastomas, we used a previously established cell line derived from external granule layer stem cells transduced with activated c-myc (NSC-M). These immortalized NSCs were able to differentiate into neurons in vitro. In contrast, when the cells were engineered to express a doxycycline-regulated REST/NRSF transgene (NSC-M-R), they no longer underwent terminal neuronal differentiation in vitro. When injected into intracranial locations in mice, the NSC-M cells did not form tumors either in the cerebellum or in the cerebral cortex. In contrast, the NSC-M-R cells did produce tumors in the cerebellum, the site of human medulloblastoma formation, but not when injected into the cerebral cortex. Furthermore, the NSC-M-R tumors were blocked from terminal neuronal differentiation. In addition, countering REST/NRSF function blocked the tumorigenic potential of NSC-M-R cells. To our knowledge, this is the first study in which abnormal expression of a sequence specific DNA-binding transcriptional repressor has been shown to contribute directly to brain tumor formation. Our findings indicate that abnormal expression of REST/NRSF and Myc in NSCs causes cerebellum-specific tumors by blocking neuronal differentiation and thus maintaining the "stemness" of these cells. Furthermore, these results suggest that such a mechanism plays a role in the formation of human medulloblastoma. PMID- 16478989 TI - Regulation of nuclear translocation of extracellular signal-regulated kinase 5 by active nuclear import and export mechanisms. AB - Extracellular signal-regulated kinase 5 (ERK5), a member of the mitogen-activated protein kinase family, plays an important role in growth factor signaling to the nucleus. However, molecular mechanisms regulating subcellular localization of ERK5 have remained unclear. Here, we show that nucleocytoplasmic shuttling of ERK5 is regulated by a bipartite nuclear localization signal-dependent nuclear import mechanism and a CRM1-dependent nuclear export mechanism. Our results show that the N-terminal half of ERK5 binds to the C-terminal half and that this binding is necessary for nuclear export of ERK5. They further show that the activating phosphorylation of ERK5 by MEK5 results in the dissociation of the binding between the N- and C-terminal halves and thus inhibits nuclear export of ERK5, causing its nuclear import. These results reveal the mechanism by which the activating phosphorylation of ERK5 induces its nuclear import and suggest a novel example of a phosphorylation-dependent control mechanism for nucleocytoplasmic shuttling of proteins. PMID- 16478991 TI - Targeted disruption of fibulin-4 abolishes elastogenesis and causes perinatal lethality in mice. AB - Elastic fibers provide tissues with elasticity which is critical to the function of arteries, lungs, skin, and other dynamic organs. Loss of elasticity is a major contributing factor in aging and diseases. However, the mechanism of elastic fiber development and assembly is poorly understood. Here, we show that lack of fibulin-4, an extracellular matrix molecule, abolishes elastogenesis. fibulin-4-/ mice generated by gene targeting exhibited severe lung and vascular defects including emphysema, artery tortuosity, irregularity, aneurysm, rupture, and resulting hemorrhages. All the homozygous mice died perinatally. The earliest abnormality noted was a uniformly narrowing of the descending aorta in fibulin-4 /- embryos at embryonic day 12.5 (E12.5). Aorta tortuosity and irregularity became noticeable at E15.5. Histological analysis demonstrated that fibulin-4-/- mice do not develop intact elastic fibers but contain irregular elastin aggregates. Electron microscopy revealed that the elastin aggregates are highly unusual in that they contain evenly distributed rod-like filaments, in contrast to the amorphous appearance of normal elastic fibers. Desmosine analysis indicated that elastin cross-links in fibulin-4-/- tissues were largely diminished. However, expression of tropoelastin or lysyl oxidase mRNA was unaffected in fibulin-4-/- mice. In addition, fibulin-4 strongly interacts with tropoelastin and colocalizes with elastic fibers in culture. These results demonstrate that fibulin-4 plays an irreplaceable role in elastogenesis. PMID- 16478990 TI - Active role for nibrin in the kinetics of atm activation. AB - The Atm protein kinase is central to the DNA double-strand break response in mammalian cells. After irradiation, dimeric Atm undergoes autophosphorylation at Ser 1981 and dissociates into active monomers. Atm activation is stimulated by expression of the Mre11/Rad50/nibrin complex. Previously, we showed that a C terminal fragment of nibrin, containing binding sites for both Mre11 and Atm, was sufficient to provide this stimulatory effect in Nijmegen breakage syndrome (NBS) cells. To discriminate whether nibrin's role in Atm activation is to bind and translocate Mre11/Rad50 to the nucleus or to interact directly with Atm, we expressed an Mre11 transgene with a C-terminal NLS sequence in NBS fibroblasts. The Mre11-NLS protein complexed with Rad50, localized to the nucleus in NBS fibroblasts, and associated with chromatin. However, Atm autophosphorylation was not stimulated in cells expressing Mre11-NLS, nor were downstream Atm targets phosphorylated. To determine whether nibrin-Atm interaction is necessary to stimulate Atm activation, we expressed nibrin transgenes lacking the Atm binding domain in NBS fibroblasts. The nibrin DeltaAtm protein interacted with Mre11/Rad50; however, Atm autophosphorylation was dramatically reduced after irradiation in NBS cells expressing the nibrin DeltaAtm transgenes relative to wild-type nibrin. These results indicate that nibrin plays an active role in Atm activation beyond translocating Mre11/Rad50 to the nucleus and that this function requires nibrin-Atm interaction. PMID- 16478992 TI - Requirement of fission yeast Cid14 in polyadenylation of rRNAs. AB - Polyadenylation in eukaryotes is conventionally associated with increased nuclear export, translation, and stability of mRNAs. In contrast, recent studies suggest that the Trf4 and Trf5 proteins, members of a widespread family of noncanonical poly(A) polymerases, share an essential function in Saccharomyces cerevisiae that involves polyadenylation of nuclear RNAs as part of a pathway of exosome-mediated RNA turnover. Substrates for this pathway include aberrantly modified tRNAs and precursors of snoRNAs and rRNAs. Here we show that Cid14 is a Trf4/5 functional homolog in the distantly related fission yeast Schizosaccharomyces pombe. Unlike trf4 trf5 double mutants, cells lacking Cid14 are viable, though they suffer an increased frequency of chromosome missegregation. The Cid14 protein is constitutively nucleolar and is required for normal nucleolar structure. A minor population of polyadenylated rRNAs was identified. These RNAs accumulated in an exosome mutant, and their presence was largely dependent on Cid14, in line with a role for Cid14 in rRNA degradation. Surprisingly, both fully processed 25S rRNA and rRNA processing intermediates appear to be channeled into this pathway. Our data suggest that additional substrates may include the mRNAs of genes involved in meiotic regulation. Polyadenylation-assisted nuclear RNA turnover is therefore likely to be a common eukaryotic mechanism affecting diverse biological processes. PMID- 16478993 TI - GCUNC-45 is a novel regulator for the progesterone receptor/hsp90 chaperoning pathway. AB - The hsp90 chaperoning pathway is a multiprotein system that is required for the production or activation of many cell regulatory proteins, including the progesterone receptor (PR). We report here the identity of GCUNC-45 as a novel modulator of PR chaperoning by hsp90. GCUNC-45, previously implicated in the activities of myosins, can interact in vivo and in vitro with both PR-A and PR-B and with hsp90. Overexpression and knockdown experiments show GCUNC-45 to be a positive factor in promoting PR function in the cell. GCUNC-45 binds to the ATP binding domain of hsp90 to prevent the activation of its ATPase activity by the cochaperone Aha1. This effect limits PR chaperoning by hsp90, but this can be reversed by FKBP52, a cochaperone that is thought to act later in the pathway. These findings reveal a new cochaperone binding site near the N terminus of hsp90, add insight on the role of FKBP52, and identify GCUNC-45 as a novel regulator of the PR signaling pathway. PMID- 16478994 TI - Autoregulation of ribosome biosynthesis by a translational response in fission yeast. AB - Maintaining the appropriate balance between the small and large ribosomal subunits is critical for translation and cell growth. We previously identified the 40S ribosomal protein S2 (rpS2) as a substrate of the protein arginine methyltransferase 3 (RMT3) and reported a misregulation of the 40S/60S ratio in rmt3 deletion mutants of Schizosaccharomyces pombe. For this study, using DNA microarrays, we have investigated the genome-wide biological response of rmt3 null cells to this ribosomal subunit imbalance. Whereas little change was observed at the transcriptional level, a number of genes showed significant alterations in their polysomal-to-monosomal ratios in rmt3Delta mutants. Importantly, nearly all of the 40S ribosomal protein-encoding mRNAs showed increased ribosome density in rmt3 disruptants. Sucrose gradient analysis also revealed that the ribosomal subunit imbalance detected in rmt3-null cells is due to a deficit in small-subunit levels and can be rescued by rpS2 overexpression. Our results indicate that rmt3-null fission yeast compensate for the reduced levels of small ribosomal subunits by increasing the ribosome density, and likely the translation efficiency, of 40S ribosomal protein-encoding mRNAs. Our findings support the existence of autoregulatory mechanisms that control ribosome biosynthesis and translation as an important layer of gene regulation. PMID- 16478995 TI - Functional evolution of the photolyase/cryptochrome protein family: importance of the C terminus of mammalian CRY1 for circadian core oscillator performance. AB - Cryptochromes (CRYs) are composed of a core domain with structural similarity to photolyase and a distinguishing C-terminal extension. While plant and fly CRYs act as circadian photoreceptors, using the C terminus for light signaling, mammalian CRY1 and CRY2 are integral components of the circadian oscillator. However, the function of their C terminus remains to be resolved. Here, we show that the C-terminal extension of mCRY1 harbors a nuclear localization signal and a putative coiled-coil domain that drive nuclear localization via two independent mechanisms and shift the equilibrium of shuttling mammalian CRY1 (mCRY1)/mammalian PER2 (mPER2) complexes towards the nucleus. Importantly, deletion of the complete C terminus prevents mCRY1 from repressing CLOCK/BMAL1 mediated transcription, whereas a plant photolyase gains this key clock function upon fusion to the last 100 amino acids of the mCRY1 core and its C terminus. Thus, the acquirement of different (species-specific) C termini during evolution not only functionally separated cryptochromes from photolyase but also caused diversity within the cryptochrome family. PMID- 16478996 TI - The insulin-like growth factor I receptor is required for Akt activation and suppression of anoikis in cells transformed by the ETV6-NTRK3 chimeric tyrosine kinase. AB - Signaling through the insulin-like growth factor I receptor (IGF-IR) axis is essential for transformation by many dominantly acting oncoproteins. However, the mechanism by which IGF-IR contributes to oncogenesis remains unknown. To examine this, we compared transformation properties of the oncogenic ETV6-NTRK3 (EN) chimeric tyrosine kinase in IGF-IR-null R- mouse embryo fibroblasts with R- cells engineered to reexpress IGF-IR (R+ cells). We previously showed that R- cells expressing EN (R- EN cells) are resistant to transformation but that transformation is restored in R+ cells. We now show that while R- EN cells have intact Ras-extracellular signal-regulated kinase signaling and cell cycle progression, they are defective in phosphatidylinositol-3-kinase (PI3K)-Akt activation and undergo detachment-induced apoptosis (anoikis) under anchorage independent conditions. In contrast, R+ cells expressing EN (R+ EN cells) suppress anoikis and are fully transformed. The requirement for IGF-IR in R- EN cells is overcome by ectopic expression of either activated Akt or a membrane targeted form of EN. Moreover, compared to R- EN cells, R+ EN cells show a dramatic increase in membrane localization of insulin receptor substrate 1 (IRS 1) in association with EN. Since EN is known to bind IRS-1 as an adaptor protein, our findings suggest that IGF-IR may function to localize EN/IRS-1 complexes to cell membranes, in turn facilitating PI3K-Akt activation and suppression of anoikis. PMID- 16478997 TI - Sp1 deacetylation induced by phorbol ester recruits p300 to activate 12(S) lipoxygenase gene transcription. AB - We previous reported that Sp1 recruits c-Jun to the promoter of the 12(S) lipoxygenase gene in 12-myristate 13-acetate-treated cells. We now show that Sp1 that recruited HDAC1 to the Sp1/cJun complex was constitutively acetylated when cells were exposed to phorbol 12-myristate 13-acetate (PMA) (3 h). Prolonged stimulation of the cells with PMA (9 h), however, caused the dissociation of histone deacetylase 1 (HDAC1) and the deacetylation of Sp1, with the latter being able to recruit p300 that in turn caused the acetylation and dissociation of histone 3, thus enhancing the expression of 12(S)-lipoxygenase. We also overexpressed an Sp1 mutant (K703/A, lacking acetylation sites) in the cell and found that cells recruited more p300 and expressed more 12(S)-lipoxygenase. Taken together, our results indicated that Sp1 recruits HDAC1 together with c-Jun to the gene promoter, followed by deacetylation of Sp1 upon PMA treatment. p300 is then recruited to the gene promoter through the interaction with deacetylated Sp1 to acetylate histone 3, leading to the enhancement of the expression of 12(S) lipoxygenase. PMID- 16478998 TI - SUMO modification of human XRCC4 regulates its localization and function in DNA double-strand break repair. AB - The nonhomologous end-joining (NHEJ) pathway is responsible for rejoining the majority of double-strand breaks in mammalian cells, including the programmed breaks introduced by V(D)J recombination. The regulation of the enzymatic activities associated with this recombination pathway is still largely unknown. Here we report that human XRCC4 (for X-ray cross-complementation group 4), a protein essential for NHEJ, is subject to posttranslational protein modification. The modifier peptide, SUMO, can be added to XRCC4 both in vitro and in vivo. The site of modification is mapped to lysine 210 by using specific mutagenesis. A protein mutated such that it cannot be SUMOylated remains localized in the cytoplasm rather than accumulating in the nucleus. Cells expressing only the mutated protein are radiation sensitive and fail to complete V(D)J recombination. Genetic fusion of the SUMO sequence to the C terminus of the mutant restores nuclear localization and radiation resistance. The modification may serve a regulatory role. Our finding fits with an emerging literature associating SUMO modification with the control of the repair and recombination associated with DNA breaks. PMID- 16478999 TI - Nuclear assembly of UGA decoding complexes on selenoprotein mRNAs: a mechanism for eluding nonsense-mediated decay? AB - Recoding of UGA from a stop codon to selenocysteine poses a dilemma for the protein translation machinery. In eukaryotes, two factors that are crucial to this recoding process are the mRNA binding protein of the Sec insertion sequence, SBP2, and the specialized elongation factor, EFsec. We sought to determine the subcellular localization of these selenoprotein synthesis factors in mammalian cells and thus gain insight into how selenoprotein mRNAs might circumvent nonsense-mediated decay. Intriguingly, both EFsec and SBP2 localization differed depending on the cell line but significant colocalization of the two proteins was observed in cells where SBP2 levels were detectable. We identify functional nuclear localization and export signals in both proteins, demonstrate that SBP2 undergoes nucleocytoplasmic shuttling, and provide evidence that SBP2 levels and localization may influence EFsec localization. Our results suggest a mechanism for the nuclear assembly of the selenocysteine incorporation machinery that could allow selenoprotein mRNAs to circumvent nonsense-mediated decay, thus providing new insights into the mechanism of selenoprotein translation. PMID- 16479001 TI - gamma-Parvin is dispensable for hematopoiesis, leukocyte trafficking, and T-cell dependent antibody response. AB - Integrins regulate cell behavior through the assembly of multiprotein complexes at the site of cell adhesion. Parvins are components of such a multiprotein complex. They consist of three members (alpha-, beta-, and gamma-parvin), form a functional complex with integrin-linked kinase (ILK) and PINCH, and link integrins to the actin cytoskeleton. Whereas alpha- and beta-parvins are widely expressed, gamma-parvin has been reported to be expressed in hematopoietic organs. In the present study, we report the expression pattern of the parvins in hematopoietic cells and the phenotypic analysis of gamma-parvin-deficient mice. Whereas alpha-parvin is not expressed in hematopoietic cells, beta-parvin is only found in myeloid cells and gamma-parvin is present in both cells of the myeloid and lymphoid lineages, where it binds ILK. Surprisingly, loss of gamma-parvin expression had no effect on blood cell differentiation, proliferation, and survival and no consequence for the T-cell-dependent antibody response and lymphocyte and dendritic cell migration. These data indicate that despite the high expression of gamma-parvin in hematopoietic cells it must play a more subtle role for blood cell homeostasis. PMID- 16479000 TI - Lipid raft targeting of hematopoietic protein tyrosine phosphatase by protein kinase C theta-mediated phosphorylation. AB - Protein kinase C theta (PKC theta) is unique among PKC isozymes in its translocation to the center of the immune synapse in T cells and its unique downstream signaling. Here we show that the hematopoietic protein tyrosine phosphatase (HePTP) also accumulates in the immune synapse in a PKC theta dependent manner upon antigen recognition by T cells and is phosphorylated by PKC theta at Ser-225, which is required for lipid raft translocation. Immune synapse translocation was completely absent in antigen-specific T cells from PKC theta-/- mice. In intact T cells, HePTP-S225A enhanced T-cell receptor (TCR)-induced NFAT/AP-1 transactivation, while the acidic substitution mutant was as efficient as wild-type HePTP. We conclude that HePTP is phosphorylated in the immune synapse by PKC theta and thereby targeted to lipid rafts to temper TCR signaling. This represents a novel mechanism for the active immune synapse recruitment and activation of a phosphatase in TCR signaling. PMID- 16479002 TI - Disruption of SLP-76 interaction with Gads inhibits dynamic clustering of SLP-76 and FcepsilonRI signaling in mast cells. AB - We developed a confocal real-time imaging approach that allows direct observation of the subcellular localization pattern of proteins involved in proximal FcepsilonRI signaling in RBL cells and primary bone marrow-derived mast cells. The adaptor protein Src homology 2 (SH2) domain-containing leukocyte phosphoprotein of 76 kDa (SLP-76) is critical for FcepsilonRI-induced calcium flux, degranulation, and cytokine secretion. In this study, we imaged SLP-76 and found it in the cytosol of unstimulated cells. Upon FcepsilonRI cross-linking, SLP-76 translocates to the cell membrane, forming clusters that colocalize with the FcepsilonRI, the tyrosine kinase Syk, the adaptor LAT, and phosphotyrosine. The disruption of the SLP-76 interaction with its constitutive binding partner, Gads, through the mutation of SLP-76 or the expression of the Gads-binding region of SLP-76, inhibits the translocation and clustering of SLP-76, suggesting that the interaction of SLP-76 with Gads is critical for appropriate subcellular localization of SLP-76. We further demonstrated that the expression of the Gads binding region of SLP-76 in bone marrow-derived mast cells inhibits FcepsilonRI induced calcium flux, degranulation, and cytokine secretion. These studies revealed, for the first time, that SLP-76 forms signaling clusters following FcepsilonRI stimulation and demonstrated that the Gads-binding region of SLP-76 regulates clustering of SLP-76 and FcepsilonRI-induced mast cell responses. PMID- 16479003 TI - Induction of CAF-1 expression in response to DNA strand breaks in quiescent human cells. AB - Genome stability in eukaryotic cells is maintained through efficient DNA damage repair pathways, which have to access and utilize chromatin as their natural template. Here we investigate the role of chromatin assembly factor 1 (CAF-1) and its interacting protein, PCNA, in the response of quiescent human cells to DNA double-strand breaks (DSBs). The expression of CAF-1 and PCNA is dramatically induced in quiescent cells upon the generation of DSBs by the radiomimetic drug bleocin (a bleomycin compound) or by ionizing radiation. This induction depends on DNA-PK. CAF-1 and PCNA are recruited to damaged chromatin undergoing DNA repair of single- and double-strand DNA breaks by the base excision repair and nonhomologous end-joining pathways, respectively, in the absence of extensive DNA synthesis. CAF-1 prepared from repair-proficient quiescent cells after induction by bleocin mediates nucleosome assembly in vitro. Depletion of CAF-1 by RNA interference in bleocin-treated quiescent cells in vivo results in a significant loss of cell viability and an accumulation of DSBs. These results support a novel and essential role for CAF-1 in the response of quiescent human cells to DSBs, possibly by reassembling chromatin following repair of DNA strand breaks. PMID- 16479005 TI - Telomere position effect and silencing of transgenes near telomeres in the mouse. AB - Reversible transcriptional silencing of genes located near telomeres, termed the telomere position effect (TPE), is well characterized in Saccharomyces cerevisiae. TPE has also been observed in human tumor cell lines, but its function remains unknown. To investigate TPE in normal mammalian cells, we developed clones of mouse embryonic stem (ES) cells that contain single-copy marker genes integrated adjacent to different telomeres. Analysis of these telomeric transgenes demonstrated that they were expressed at very low levels compared to the same transgenes integrated at interstitial sites. Similar to the situation in yeast, but in contrast to studies with human tumor cell lines, TPE in mouse ES cells was not reversed with trichostatin A. Prolonged culturing without selection resulted in extensive DNA methylation and complete silencing of telomeric transgenes, which could be reversed by treatment with 5-azacytidine. Thus, complete silencing of the telomeric transgenes appears to involve a two step process in which the initial repression is reinforced by DNA methylation. Extensive methylation of the telomeric transgenes was also observed in various tissues and embryonic fibroblasts isolated from transgenic mice. In contrast, telomeric transgenes were not silenced in ES cell lines isolated from 3-day-old preimplantation embryos, consistent with the hypothesis that TPE plays a role in the development of the embryo. PMID- 16479004 TI - Mammalian Rad9 plays a role in telomere stability, S- and G2-phase-specific cell survival, and homologous recombinational repair. AB - The protein products of several rad checkpoint genes of Schizosaccharomyces pombe (rad1+, rad3+, rad9+, rad17+, rad26+, and hus1+) play crucial roles in sensing changes in DNA structure, and several function in the maintenance of telomeres. When the mammalian homologue of S. pombe Rad9 was inactivated, increases in chromosome end-to-end associations and frequency of telomere loss were observed. This telomere instability correlated with enhanced S- and G2-phase-specific cell killing, delayed kinetics of gamma-H2AX focus appearance and disappearance, and reduced chromosomal repair after ionizing radiation (IR) exposure, suggesting that Rad9 plays a role in cell cycle phase-specific DNA damage repair. Furthermore, mammalian Rad9 interacted with Rad51, and inactivation of mammalian Rad9 also resulted in decreased homologous recombinational (HR) repair, which occurs predominantly in the S and G2 phases of the cell cycle. Together, these findings provide evidence of roles for mammalian Rad9 in telomere stability and HR repair as a mechanism for promoting cell survival after IR exposure. PMID- 16479006 TI - Mice lacking the nuclear pore complex protein ALADIN show female infertility but fail to develop a phenotype resembling human triple A syndrome. AB - Triple A syndrome is a human autosomal recessive disorder characterized by adrenal insufficiency, achalasia, alacrima, and neurological abnormalities affecting the central, peripheral, and autonomic nervous systems. In humans, this disease is caused by mutations in the AAAS gene, which encodes ALADIN, a protein that belongs to the family of WD-repeat proteins and localizes to nuclear pore complexes. To analyze the function of the gene in the context of the whole organism and in an attempt to obtain an animal model for human triple A syndrome, we generated mice lacking a functional Aaas gene. The Aaas-/- animals were found to be externally indistinguishable from their wild-type littermates, although their body weight was on the average lower than that of wild-type mice. Histological analysis of various tissues failed to reveal any differences between Aaas-/- and wild-type mice. Aaas-/- mice exhibit unexpectedly mild abnormal behavior and only minor neurological deficits. Our data show that the lack of ALADIN in mice does not lead to a triple A syndrome-like disease. Thus, in mice either the function of ALADIN differs from that in humans, its loss can be readily compensated for, or additional factors, such as environmental conditions or genetic modifiers, contribute to the disease. PMID- 16479007 TI - Inquiring into the differential action of interferons (IFNs): an IFN-alpha2 mutant with enhanced affinity to IFNAR1 is functionally similar to IFN-beta. AB - Alpha and beta interferons (IFN-alpha and IFN-beta) are multifunctional cytokines that exhibit differential activities through a common receptor composed of the subunits IFNAR1 and IFNAR2. Here we combined biophysical and functional studies to explore the mechanism that allows the alpha and beta IFNs to act differentially. For this purpose, we have engineered an IFN-alpha2 triple mutant termed the HEQ mutant that mimics the biological properties of IFN-beta. Compared to wild-type (wt) IFN-alpha2, the HEQ mutant confers a 30-fold higher binding affinity towards IFNAR1, comparable to that measured for IFN-beta, resulting in a much higher stability of the ternary complex as measured on model membranes. The HEQ mutant, like IFN-beta, promotes a differentially higher antiproliferative effect than antiviral activity. Both bring on a down-regulation of the IFNAR2 receptor upon induction, confirming an increased ternary complex stability of the plasma membrane. Oligonucleotide microarray experiments showed similar gene transcription profiles induced by the HEQ mutant and IFN-beta and higher levels of gene induction or repression than those for wt IFN-alpha2. Thus, we show that the differential activities of IFN-beta are directly related to the binding affinity for IFNAR1. Conservation of the residues mutated in the HEQ mutant within IFN-alpha subtypes suggests that IFN-alpha has evolved to bind IFNAR1 weakly, apparently to sustain differential levels of biological activities compared to those induced by IFN-beta. PMID- 16479008 TI - Regulation of sprouty stability by Mnk1-dependent phosphorylation. AB - Sprouty (Spry) proteins are negative feedback modulators of receptor tyrosine kinase pathways in Drosophila melanogaster and mammals. Mammalian Spry proteins have been shown to undergo tyrosine and serine phosphorylation in response to growth factor stimulation. While several studies have addressed the function of tyrosine phosphorylation of Spry, little is known about the significance of Spry serine phosphorylation. Here we identify mitogen-activated protein kinase interacting kinase 1 (Mnk1) as the kinase that phosphorylates human Spry2 (hSpry2) on serines 112 and 121. Mutation of these serine residues to alanine or inhibition of Mnk1 activity increases the rate of ligand-induced degradation of hSpry2. Conversely, enhancement of serine phosphorylation achieved through either the inhibition of cellular phosphatases or the expression of active Mnk1 results in the stabilization of hSpry2. Previous studies have shown that growth factor stimulation induces the proteolytic degradation of hSpry2 by stimulating tyrosine phosphorylation on hSpry2, which in turn promotes c-Cbl binding and polyubiquitination. A mutant of hSpry2 that is deficient in serine phosphorylation displays enhanced tyrosine phosphorylation and c-Cbl binding, indicating that serine phosphorylation stabilizes hSpry2 by exerting an antagonistic effect on tyrosine phosphorylation. Moreover, loss of serine phosphorylation and the resulting enhanced degradation of hSpry2 impair its capacity to antagonize fibroblast growth factor-induced extracellular signal regulated kinase activation. Our results imply that Mnk1-mediated serine phosphorylation of hSpry2 constitutes a regulatory mechanism to extend the temporal range of Spry activity. PMID- 16479009 TI - Androgen and its receptor promote Bax-mediated apoptosis. AB - Androgen and its receptor (AR) have been reported to have pro- or antiapoptotic functions. However, the underlying molecular mechanism is incompletely understood. We report here that androgen and AR promote Bax-mediated apoptosis in prostate cancer cells. UV irradiation and ectopic expression of Bax induce apoptosis in AR-positive, but not AR-negative prostate cancer cells. UV- and Bax induced apoptosis is abrogated in AR-positive cells that express small interference RNA (siRNA) of AR and is sensitized by reintroduction of AR into AR negative cells. Although AR is able to promote Bax-mediated apoptosis independently of androgen, the promotion by AR can be further potentiated by androgen via AR-dependent transcription activation. AR is essential for the translocation of Bax to mitochondria in UV- or Bax-induced apoptosis. Inhibition of Bax expression by Bax siRNA suppresses UV-induced apoptosis in AR-positive cells. In addition, introduction of AR into AR-negative prostate cancer cells upregulates expression levels of the BH3-only protein Noxa, whereas inhibition of Noxa expression reduces the promotion by AR on UV-induced apoptosis. Thus, our results reveal a novel cross talk between the androgen/AR hormonal signaling pathway and the intrinsic apoptotic death pathway that determines the sensitivity of stress-induced apoptosis in prostate cancer cells. PMID- 16479010 TI - RBP1 family proteins exhibit SUMOylation-dependent transcriptional repression and induce cell growth inhibition reminiscent of senescence. AB - The retinoblastoma binding protein 1 (RBP1) appears to be an important factor in the repression of E2F-dependent transcription by the retinoblastoma protein (pRB) family. The recent identification of the breast carcinoma associated antigen (BCAA) as an RBP1-like protein led us to investigate its biological properties and compare them to RBP1. Like RBP1, BCAA contains a carboxy-terminal R2 domain that elicits histone deacetylase (HDAC)-dependent transcriptional repression via interactions with the SAP30 subunit of the Sin3/HDAC complex. Each RBP1 family member also contains two HDAC-independent repression activities within a region termed R1, which can be subdivided into a SUMOylated moiety (R1sigma) and a predicted alpha-helical region (R1alpha). R1alpha is embedded within the ARID region and represses basal transcription only, whereas R1sigma represses both basal and activated transcription and depends on SUMOylation. Overexpression of either RBP1 or BCAA, but not the truncated BCAAMCF-7 isoform that is overexpressed in breast cancer cells, caused a profound inhibition of cell proliferation and induced expression of a senescence marker. In each case the presence of both R1 and R2 was necessary for suppression of cell growth, suggesting that both R1 and R2 transcriptional repression activities play a role in RBP1 family protein-mediated regulation of cellular proliferation. PMID- 16479012 TI - Sodium nitroprusside promotes IRP2 degradation via an increase in intracellular iron and in the absence of S nitrosylation at C178. AB - In iron-replete cells the posttranscriptional regulator IRP2 undergoes ubiquitination and proteasomal degradation. A similar response occurs in cells exposed to sodium nitroprusside (SNP), an NO-releasing drug. It has been proposed that nitroprusside ([Fe(CN)5NO]2-) fails to donate iron into cells and that it promotes IRP2 degradation via S nitrosylation at C178. This residue is located within a stretch of 73 amino acids, earlier proposed to define an iron-dependent degradation domain. Surprisingly, we show that IRP2 bearing a C178S mutation or a Delta73 deletion is sensitive to degradation not only by ferric ammonium citrate (FAC) but also by SNP. Moreover, FAC and SNP attenuate the RNA-binding activities of IRP2 and its homologue IRP1 with similar kinetics. Actinomycin D, cycloheximide, succinylacetone, and dimethyl-oxalylglycine antagonize IRP2 degradation in response to both FAC and SNP, suggesting a common mechanistic basis. IRP2 is not only sensitive to fresh, but also to photodegraded SNP and remains unaffected by S-nitrosoglutathione (GSNO), an established nitrosation agent. Importantly, both fresh and photodegraded SNP, but not GSNO, promote a >4 fold increase in the calcein-accessible labile iron pool. Collectively, these results suggest that IRP2 degradation by SNP does not require S nitrosylation but rather represents a response to iron loading. PMID- 16479011 TI - The adaptor protein Tom1L1 is a negative regulator of Src mitogenic signaling induced by growth factors. AB - The Src family of protein-tyrosine kinases (SFK) play important roles in mitogenesis and morphological changes induced by growth factors. The involved substrates are, however, ill defined. Using an antiphosphotyrosine antibody to screen tyrosine-phosphorylated cDNA expression library, we have identified Tom1L1, an adaptor protein of the Tom1 family and a novel substrate and activator of the SFK. Surprisingly, we found that Tom1L1 does not promote DNA synthesis induced by Src. Furthermore, we report that Tom1L1 negatively regulates SFK mitogenic signaling induced by platelet-derived growth factor (PDGF) through modulation of SFK-receptor association: (i) Tom1L1 inhibits DNA synthesis induced by PDGF; (ii) inhibition is overcome by c-myc expression or p53 inactivation, two regulators of SFK mitogenic function; (iii) Src or Fyn coexpression overrides Tom1L1 mitogenic activity; (iv) overexpression of the adaptor reduces Src association with the receptor; and (v) protein inactivation potentiates receptor complex formation, allowing increased SFK activation and DNA synthesis. However, Tom1L1 affects neither DNA synthesis induced by the constitutively active allele SrcY527F nor SFK-regulated actin assembly induced by PDGF. Finally, overexpressed Tom1 and Tom1L2 also associate with Src and affected mitogenic signaling in agreement with some redundancy among members of the Tom1 family. We concluded that Tom1L1 defines a novel mechanism for regulation of SFK mitogenic signaling induced by growth factors. PMID- 16479013 TI - Tolerance of Sir1p/origin recognition complex-dependent silencing for enhanced origin firing at HMRa. AB - The HMR-E silencer is a DNA element that directs the formation of silent chromatin at the HMRa locus in Saccharomyces cerevisiae. Sir1p is one of four Sir proteins required for silent chromatin formation at HMRa. Sir1p functions by binding the origin recognition complex (ORC), which binds to HMR-E, and recruiting the other Sir proteins (Sir2p to -4p). ORCs also bind to hundreds of nonsilencer positions distributed throughout the genome, marking them as replication origins, the sites for replication initiation. HMR-E also acts as a replication origin, but compared to many origins in the genome, it fires extremely inefficiently and late during S phase. One postulate to explain this observation is that ORC's role in origin firing is incompatible with its role in binding Sir1p and/or the formation of silent chromatin. Here we examined a mutant HMR-E silencer and fusions between robust replication origins and HMR-E for HMRa silencing, origin firing, and replication timing. Origin firing within HMRa and from the HMR-E silencer itself could be significantly enhanced, and the timing of HMRa replication during an otherwise normal S phase advanced, without a substantial reduction in SIR1-dependent silencing. However, although the robust origin/silencer fusions silenced HMRa quite well, they were measurably less effective than a comparable silencer containing HMR-E's native ORC binding site. PMID- 16479014 TI - The proteasome is required for rapid initiation of death receptor-induced apoptosis. AB - Due to their tremendous apoptosis-inducing potential, proteasomal inhibitors (PIs) have recently entered clinical trials. Here we show, however, that various PIs rescued proliferating tumor cells from death receptor-induced apoptosis. This protection correlated with the stabilization of X-linked IAP (XIAP) and c-FLIP and the inhibition of caspase activation. Together with the observation that PIs could not protect cells expressing XIAP or c-FLIP short interfering RNAs (siRNAs) from death receptor-induced apoptosis, our results demonstrate that PIs mediate their protective effect via the stabilization of these antiapoptotic proteins. Furthermore, we show that once these proteins were eliminated, either by long term treatment with death receptor ligands or by siRNA-mediated suppression, active caspases accumulated to an even larger extent in the presence of PIs. Together, our data support a biphasic role for the proteasome in apoptosis, as they show that its constitutive activity is crucial for the rapid initiation of the death program by eliminating antiapoptotic proteins, whereas at later stages, the proteasome acts in an antiapoptotic manner due to the proteolysis of caspases. Thus, for a successful PI-based tumor therapy, it is crucial to carefully evaluate basal proteasomal activity and the status of antiapoptotic proteins, as their PI-mediated prolonged stability might even cause adverse effects, leading to the survival of a tumor. PMID- 16479016 TI - Protein phosphatase 2A antagonizes ATM and ATR in a Cdk2- and Cdc7-independent DNA damage checkpoint. AB - We previously used a soluble cell-free system derived from Xenopus eggs to investigate the role of protein phosphatase 2A (PP2A) in chromosomal DNA replication. We found that immunodepletion of PP2A or inhibition of PP2A by okadaic acid (OA) inhibits initiation of DNA replication by preventing loading of the initiation factor Cdc45 onto prereplication complexes. Evidence was provided that PP2A counteracts an inhibitory protein kinase that phosphorylates and inactivates a crucial Cdc45 loading factor. Here, we report that the inhibitory effect of OA is abolished by caffeine, an inhibitor of the checkpoint kinases ataxia-telangiectasia mutated protein (ATM) and ataxia-telangiectasia related protein (ATR) but not by depletion of ATM or ATR from the extract. Furthermore, we demonstrate that double-strand DNA breaks (DSBs) cause inhibition of Cdc45 loading and initiation of DNA replication and that caffeine, as well as immunodepletion of either ATM or ATR, abolishes this inhibition. Importantly, the DSB-induced inhibition of Cdc45 loading is prevented by addition of the catalytic subunit of PP2A to the extract. These data suggest that DSBs and OA prevent Cdc45 loading through different pathways, both of which involve PP2A, but only the DSB induced checkpoint implicates ATM and ATR. The inhibitory effect of DSBs on Cdc45 loading does not result from downregulation of cyclin-dependent kinase 2 (Cdk2) or Cdc7 activity and is independent of Chk2. However, it is partially dependent on Chk1, which becomes phosphorylated in response to DSBs. These data suggest that PP2A counteracts ATM and ATR in a DNA damage checkpoint in Xenopus egg extracts. PMID- 16479015 TI - Interferon-inducible protein IFIXalpha1 functions as a negative regulator of HDM2. AB - The 200-amino-acid repeat (HIN-200) gene family with the hematopoietic interferon (IFN)-inducible nuclear protein encodes highly homologous proteins involved in cell growth, differentiation, autoimmunity, and tumor suppression. IFIX is the newest member of the human HIN-200 family and is often downregulated in breast tumors and breast cancer cell lines. The expression of the longest isoform of IFIX gene products, IFIXalpha1, is associated with growth inhibition, suppression of transformation, and tumorigenesis. However, the mechanism underlying the tumor suppression activity of IFIXalpha1 is not well understood. Here, we show that IFIXalpha1 downregulates HDM2, a principal negative regulator of p53, at the posttranslational level. IFIXalpha1 destabilizes HDM2 protein and promotes its ubiquitination. The E3 ligase activity of HDM2 appears to be required for this IFIXalpha1 effect. Importantly, HDM2 downregulation is required for the IFIXalpha1-mediated increase of p53 protein levels, transcriptional activity, and nuclear localization, suggesting that IFIXalpha1 positively regulates p53 by acting as a negative regulator of HDM2. We found that IFIXalpha1 interacts with HDM2. Interestingly, the signature motif of the HIN-200 gene family, i.e., the 200-amino-acid HIN domain of IFIXalpha1, is sufficient not only for binding HDM2 but also for downregulating it, leading to p53 activation. Finally, we show that IFIX mediates HDM2 downregulation in an IFN-inducible system. Together, these results suggest that IFIXalpha1 functions as a tumor suppressor by repressing HDM2 function. PMID- 16479017 TI - Alpha-fetoprotein controls female fertility and prenatal development of the gonadotropin-releasing hormone pathway through an antiestrogenic action. AB - It has been shown previously that female mice homozygous for an alpha-fetoprotein (AFP) null allele are sterile as a result of anovulation, probably due to a defect in the hypothalamic-pituitary axis. Here we show that these female mice exhibit specific anomalies in the expression of numerous genes in the pituitary, including genes involved in the gonadotropin-releasing hormone pathway, which are underexpressed. In the hypothalamus, the gonadotropin-releasing hormone gene, Gnrh1, was also found to be down-regulated. However, pituitary gene expression could be normalized and fertility could be rescued by blocking prenatal estrogen synthesis using an aromatase inhibitor. These results show that AFP protects the developing female brain from the adverse effects of prenatal estrogen exposure and clarify a long-running debate on the role of this fetal protein in brain sexual differentiation. PMID- 16479019 TI - [Neuropsychological assessment of the prefrontal cortex in major depressive disorder]. AB - The pathology of the prefrontal cortex (PFC) may play an important role in the development of the symptoms of major depressive disorder. In this study, the authors used the Wisconsin Card Sorting Test (WCST) and the Iowa Gambling Test (IGT) to investigate PFC functions in depression. The WCST investigates cognitive set-shifting abilities, whereas the IGT is sensitive for the cumulative effect of reward and punishment on decision-making. Participants were 20 patients with DSM IV major depressive disorder and 20 age-, gender-, and education-matched healthy control subjects. The depressed patients showed significant impairment in both tests, but the WCST and IGT scores did not correlate. There was no significant correlation between the test results and the severity of depressive and anxiety symptoms. Our results suggest a global impairment of the PFC in depression, which includes the dorsolateral and ventromedial regions. PMID- 16479018 TI - Kdo2-Lipid A of Escherichia coli, a defined endotoxin that activates macrophages via TLR-4. AB - The LIPID MAPS Consortium (www.lipidmaps.org) is developing comprehensive procedures for identifying all lipids of the macrophage, following activation by endotoxin. The goal is to quantify temporal and spatial changes in lipids that occur with cellular metabolism and to develop bioinformatic approaches that establish dynamic lipid networks. To achieve these aims, an endotoxin of the highest possible analytical specification is crucial. We now report a large-scale preparation of 3-deoxy-D-manno-octulosonic acid (Kdo)(2)-Lipid A, a nearly homogeneous Re lipopolysaccharide (LPS) sub-structure with endotoxin activity equal to LPS. Kdo(2)-Lipid A was extracted from 2 kg cell paste of a heptose deficient Escherichia coli mutant. It was purified by chromatography on silica, DEAE-cellulose, and C18 reverse-phase resin. Structure and purity were evaluated by electrospray ionization/mass spectrometry, liquid chromatography/mass spectrometry and (1)H-NMR. Its bioactivity was compared with LPS in RAW 264.7 cells and bone marrow macrophages from wild-type and toll-like receptor 4 (TLR-4) deficient mice. Cytokine and eicosanoid production, in conjunction with gene expression profiling, were employed as readouts. Kdo(2)-Lipid A is comparable to LPS by these criteria. Its activity is reduced by >10(3) in cells from TLR-4 deficient mice. The purity of Kdo(2)-Lipid A should facilitate structural analysis of complexes with receptors like TLR-4/MD2. PMID- 16479020 TI - [Hungarian adaptation of the Treatment Demand Indicator Protocol (TDI). Results of a multicenter study]. AB - Accession to the European Union (EU) requires a revision of reporting system(s) and the adaptation of EU compliant measurement tools from the Hungarian side. In this study we try to monitor the Hungarian adaptation process of TDI regarding the European developments in this issue. The second goal of this study was to report on the reliability data of TDI which was assessed in Europe by our team first. 209 users of illicit drugs were sampled. The language adaptation needed a multi-stage translation process. The interrater and test retest reliability of TDI was found to be good. Inconsistency between related questions was not found. The reliability and validity indicators of the TDI in Hungary were acceptable. The protocol is easily applicable (5 minutes) and training is not needed. PMID- 16479022 TI - [Symptom specificity of adolescents with self-injurious behavior]. AB - As an introductory part of the paper, authors give a short overview of existing results in the literature related to self-injurious behaviour and adolescents' deliberate self-harm. In their own random sample study, authors organized a self report screen (provincial town, 3 educational facilities, 470 pupils aged between 14 and 18 years) by means of the translated version of Ottawa Self Injury Inventory (OSI) used widely in community-based studies in Canada. The Beck Depression Inventory was introduced to measure the key symptoms of depression among youngsters. 26 youngsters were found to have had at least one self injurious action in their life-time. The authors describe the characteristics of these subjects on the basis of symptom occurrence statistics. Although the depressive symptoms have an expected correlation with the self-injurious ideas, depression does not seem to have the same relationships with the actual self-harm action. The authors attempt to give an explanation of this contradiction. PMID- 16479021 TI - [Interaction patterns in male and female suicide attempters]. AB - The present study aimed to assess the interaction dynamics of suicidal patients' close relationships using the Consensus Rorschach method. Our sample consisted of 24 male and 8 female patients and their spouses as well as 30 control couples. We have analysed the couples' behavior in situations where communication difficulties ("communication crisis") have arisen, comparing communication of female and male patients to controls. Results have shown that couples with suicide history expressed more passive and passive emotional messages, with evident signs of problem solving difficulties. Male patients frequently expressed helplessness, while female patients showed poor emotional control. CONCLUSIONS: difficulties in communication and conflict management as well as lack of empathy and perspective taking observed in the relationships of these couples can be explained by disturbances in the bonding process (probably both in patients and partners); which indicates the need for individual and family therapy. PMID- 16479023 TI - [The effect of public holidays on the suicide drive (frequency) in Hungary (1970 2002)]. AB - INTRODUCTION: The aim of the study was to examine the relationship between suicide frequency and the days of the week and major public holidays in Hungary (1970-2002). The analysis was based on data of approximately 140,000 suicides. The authors intended to examine whether the "broken promise effect" (Gabennesch), which is cited with increasing frequency in the literature, is valid also in Hungary or not. RESULTS: Suicides occur most frequently on Monday and least on weekends for both males and females. On the days of Christmas and Easter the suicide drive has decreased among males only. The first day of the year is associated with a large increase of suicide risk in both genders, while there appears to be no change in suicide-frequency on the public holidays of 1st May and 20th August. CONCLUSIONS: The findings of our study confirm the validity of the "broken-promise effect" theory also under Hungarian circumstances. It is a very important to know that suicides do not show an increase on and before weekend and public holidays, but after the following day(s). This finding can be an important aspect when preventive measures are adopted. PMID- 16479024 TI - The effect of vitamin D as supplementary treatment in patients with moderately advanced pulmonary tuberculous lesion. AB - AIM: to compare the vitamin D group of pulmonary tuberculosis patients with a placebo group in terms of clinical improvement, nutritional status, sputum conversion, and radiological improvement. METHODS: sixty seven tuberculosis patient visiting the Pulmonary Clinic, of Cipto Mangunkusumo Hospital, Jakarta, from January 1st to August 31st, 2001 were included in this study. The subjects were randomised to receive vitamin D (0.25 mg/day) or placebo in a double blind method, during the 6th initial week of Tb treatment. The rate of sputum conversion, complete blood counts, blood chemistry as well as radiologic examination were evaluated. RESULTS: there were more male patients than females (39:28), 78.7% were in the productive age group, 71.6% had low nutritional status, 62.4% with low education level, and 67.2% with low income. One hundred percent of the vitamin D group and only 76.7% of the placebo group had sputum conversion. This difference is statistically significant (p=0.002). CONCLUSION: the sputum conversion had no correlation with the hemoglobin level, blood clotting time, calcium level, lymphocyte count, age, sex, and nutritional status. There were more subjects with radiological improvement in the vitamin D group. PMID- 16479025 TI - Neuropsychological findings in hyperthyroid patients. AB - AIM: to evaluate attention, concentration, memory changes and executive function inpatients with hyperthyroidism compared with normal thyroid function patients. METHODS: a cross sectional study was conducted in 21 patients (14F, 7M) with symptoms of sweating, sleep disturbance, fine tremor, headache, palpitation, nervousness, forgetfulness and diarrhea. Neurological examination found that 2 patients had proximal paresis and periodic paralysis. Their thyroid functions (FT4 and TSHS) were done, 13 patients were hyperthyroid, 2 patients were subclinical hyperthyroid and 6 patients were euthyroid. Neuropsychological tests such as: Mini Mental State Examination (MMSE), Trail Making Test (TMT) A and B, Continuous Performance Test (CPT), Vigilance Complex Task (VCT), Digit Forward and Backward Repetition (DFR, DBR) were performed in all of those patients. The mean age of sample was 47,3 +/- 8,45 years old and all of them had minimal 6 years education background, no history of cerebro vascular disease, brain injury or tumour or infection, epilepsy, cerebral degeneration, and taking neuroleptics or antidepressants. RESULTS: using the Kendall correlation coefficients showed that FT4 had negative correlation with VCT, CPT, DFR, DBR and positive correlation with TMT A (time) and TMT B (time and error). Study T-test showed that high level of thyroid function had significant difference in TMT A (time), TMT B (time), DBR, DFR, VCT, CPT. Fischer exact gave statistically significant in VCT and fine tremor in hyperthyroid patients (RR VCT 5.60, 95% CI 0.93-33.68, RR fine tremor 0.50, 95% CI 0.28-0.88). CONCLUSION: hyperthyroid patients had significant decrease on attention, concentration, verbal memory and executive function (working memory) compared with euthyroid patients. PMID- 16479026 TI - The role of lipid profile as a risk factor indicator for ischemic stroke at Cipto Mangunkusumo Hospital, Jakarta. AB - AIM: to determine the role of low HDL-cholesterol, and high total cholesterol, LDL-Cholesterol and triglyceride as risk factors for ischemic stroke at Dr. Cipto Mangunkusumo Hospital. METHOD: a study was conducted on 76 patients with an age range of 40-70 years. Subjects consisted of 38 post ischemic stroke patients and 38 control subjects with a diagnosis other than stroke. The study sample consisted of serum for lipid profile assessment. Total cholesterol and triglyceride were assessed using enzymatic method, while HDL-cholesterol and LDL cholesterol using direct homogenous enzymatic method. Statistical analysis was performed using chi-square and multivariate analysis using logistic regression. RESULTS: low HDL-cholesterol was found in ischemic stroke patients and demonstrated a significant difference compared to control subjects (p<0.05). The results of total cholesterol, triglyceride, LDL-cholesterol did not demonstrate a significant difference. The odds ratio (3.09; CI 95%: 1.04; 8.73) demonstrates that low HDL-cholesterol is a risk factor for ischemic stroke. CONCLUSION: a low HDL-cholesterol level is a risk factor for ischemic stroke, with an odds ratio of 3.09, while total cholesterol, triglyceride and high LDL-cholesterol levels were not risk factors for ischemic stroke. PMID- 16479027 TI - Insulin resistance and metabolic syndrome in elderly women living in nursing homes. AB - AIM: To obtain a depiction of insulin resistance and to determine the correlation between age, body mass index, and insulin resistance, and the prevalence of metabolic syndrome in elderly women living in nursing homes. METHODS: Ninety two elderly females with an average age of 71.4 (SD 7.45) who did not suffer from chronic renal disease or chronic liver disease and were not taking corticosteroids, beta blockers, thiazides, or anti-dyslipidemic drugs, participated as research subjects. Investigated variables, comprising blood pressure, body mass index, waist circumference, laboratory measurements, lipid profile, fasting blood glucose level, and fasting insulin level, were used to determine insulin resistance according to HOMA-IR > 75 percentile. The NCEP ATP III criteria Asia Pacific modification was used to determine metabolic syndrome. RESULTS: There were twenty three subjects with insulin resistance (HOMA-IR > 2.67) for the 75th percentile. Seventeen of them (73.9%) had metabolic syndrome. However, out of the 6 subjects (26.1%) without metabolic syndrome, 5 subjects had at least one component of metabolic syndrome. There was no correlation between age and insulin resistance. However, there was a correlation between BMI and insulin resistance (p<0.017). There were 53 elderly females (57.6%) with metabolic syndrome; the most common metabolic syndrome component being hypertension (79.3%), followed by HDL hypocholesterolemia (55.4%), and central obesity (53%). As many as 32.1% of subjects with metabolic syndrome also demonstrated insulin resistance. CONCLUSION: The value of insulin resistance was 2.67 (HOMA-IR cut-off > 75 percentile). Subjects with insulin resistance had at least one component of metabolic syndrome. A high prevalence (57.6%) of metabolic syndrome in elderly females living in a nursing home was found. PMID- 16479028 TI - Is there a relationship between serum C-reactive protein level and dyslipidaemia in systemic lupus erythematosus? AB - AIM: to look for the association between CRP and dyslipidaemia in SLE. METHODS: seventy six patients fulfilled the American College of Rheumatology criteria for classification of Systemic Lupus Erythematosus (SLE) (revised in 1997) were enrolled in our study. Clinical and laboratory measures included complete history and physical examination, determination of serum CRP level by latex agglutination test and lipid profile. Statistical significance of association was analysed by X2 test. T-test were used to compare values of each lipid component between positive and negative CRP group. All analyses were performed using the SPSS 10.0 computer software. RESULTS: the study was done in 76 (73 female and 3 male) SLE patients. Dyslipidaemia was found in 57 patients (75.0%). Hypercholesterolemia was found in 31 patients (40.8%), hypertriglyceridemia in 33 patients (43.4%), low HDL cholesterol in 19 patients (25%) and high LDL cholesterol in 28 patients (36.8%). Patients with positive CRP (66 patients) demonstrated dyslipidaemia in 49 patients (74.2%) and patients with negative CRP (10 patients) showed dyslipidaemia in 8 patients (80.0%)(P 1.0). There was no association between CRP and abnormal values of each lipid component (cholesterol, triglyceride, HDL and LDL cholesterol)(P value 0.30, 0.74, 0.43, 0.15 respectively). There was also no association between CRP and dyslipidaemia as a whole (P 1.00). The difference between serum level of each lipid component between positive and negative CRP group was also non significant (P value 0.68, 0.90, 0.96, 0.59 respectively). CONCLUSION: there was no association between CRP and dyslipidaemia in SLE patients. In the development of dyslipidaemia in SLE, factors other than inflammation should be put into consideration. PMID- 16479029 TI - Laryngeal tuberculosis: an important issue. AB - In developing countries like Indonesia, pulmonary tuberculosis still ranks among the major health problems and the prevalence of laryngeal tuberculosis are therefore still high. It is important for physicians and otolaryngologist to recognize the cardinal sign and symptoms of laryngeal tuberculosis in order to make an early diagnosis. An illustration of four cases with laryngeal tuberculosis is reported. All cases presented with hoarseness and diagnosis of laryngeal and pulmonary tuberculosis were made at ENT department. PMID- 16479030 TI - Why does diabetes mellitus increase the risk of cardiovascular disease? AB - The main etiology for mortality and a great percentage of morbidity in patients with diabetes mellitus is atherosclerosis. The pathogenesis of cardiovascular disease (CVD) in diabetes is multifactorial and can be affected by metabolic and other factors. A hypothesis for the initial lesion of atherosclerosis is endothelial dysfunction, defined pragmatically as changes in the concentration of the chemical messengers produced by the endothelial cell and/or by blunting of the nitric oxide-dependent vasodilatory response to acetylcholine or hyperemia. Endothelial dysfunction has been documented in patients with diabetes and in individuals with insulin resistance or at high risk for developing type 2 diabetes. The way endothelial function altered in diabetic patients is not yet fully understood, but the loss of normal endothelial function could be involved in the pathogenesis of diabetic angiopathy, as endothelial dysfunction is associated with diabetic microangiopathy and macroangiopathy. Factors associated with endothelial dysfunction in diabetes include activation of protein kinase C, overexpression of growth factors and/or cytokines, and oxidative stress. Changes in endothelium function may lead to the coronary artery circulation being unable to cope with the increased metabolism of myocardial muscle independently of a reduced coronary artery diameter. Finally, recent reports indicate that an improved metabolic control in diabetic patients, whatever the treatment used, is associated with near normalization or restoration of normal endothelial function. PMID- 16479031 TI - Pathogenesis and pharmacologic treatment of obesity: the role of energy regulatory mechanism. AB - Obesity has become a worldwide public health problem affecting millions of people. This is a chronic, stigmatized, and costly disease, rarely curable and is increasing in prevalence to a point today where we define obesity as an epidemic disease that not only in developed but also on developing countries. The pathogenesis of obesity is largely unknown, especially about energy regulatory mechanism that involved wide area of neuroendocrinology that is very interesting but very complex and makes internists "refuse" to learn. Obesity occurs through a longstanding imbalance between energy intake and energy expenditure, influenced by a complex biologic system that regulates appetite and adiposity. Obesity influences the pathogenesis of hypertension, type 2 diabetes, dyslipidemia, kidney, heart, and cerebrovascular disease. It is very wise for every internist to learn the pathogenesis and treatment of this worldwide diseases. Until now, the available treatments, including drugs, are palliative and are effective only while the treatment is being actively used; and besides so many side effects reported. PMID- 16479032 TI - Prognostic in sarcoma Ewing. PMID- 16479033 TI - Internet searching on internal medicine literature. AB - Internet technology for distributing information enormously supports the development of medical knowledge. Various medical resources on the internet are very helpful for the doctors' self-improvement; either on education, research, or health care application. Famous medical journals have made their online version, which can be accessed at any time and anywhere. However, there are extremely enourmous information that could be retrieved, so that the doctors should have appropriate strategy or measure to obtain the literature effectively and efficiently. PMID- 16479034 TI - The use of ACE inhibitor in cardiovascular disease. AB - Competitive inhibition of Angiotensin Converting Enzyme has an important role in cardiovascular diseases, which has been demonstrated by various extensive studies. Based on pharmacologic and mechanism of action, the pharmacodynamic of ACE-inhibitor involves some specific system, such as hemodynamic, neurohormonal, anti- proliferation and renal effect, etc. ACE-inhibitor has beneficial effect in various cardiovascular diseases such as heart failure, hypertension, systolic and dyastolic left ventricle dysfunction and acute myocardial infarction. This article will discuss advantages of its therapeutic use, the adverse effects that might occur as well as how to overcome this effect and also when it should be stopped. Extensive studies also demonstrate that therapeutic use of ACE-inhibitor will improve the quality of life and survival rate in conjunction with the decrease of morbidity and mortality. PMID- 16479035 TI - Angiographic and clinical outcome following paclitaxel-eluting stent (taxus) implantation: a single center experience. AB - Coronary stents dramatically improve acute outcomes of percutaneous coronary interventions but also induce abundant intraluminal neointimal growth. Drug eluting stents reduce intimal hyperplasia, the main cause of in-stent restenosis. The safety and beneficial effects of paclitaxel-eluting stents (Taxus) in patients treated in daily practice remains to be defined. The aim of this study was to report the late outcomes of Taxus implantation in patients with coronary artery disease. The study population consisted of 151 patients (202 stents) who had undergone coronary Taxus stent implantation between March 2003 and May 2005. Patients were eligible for enrollment if there was symptomatic coronary artery disease or positive functional testing, and angiographic evidence of single or multivessel disease with a target lesion stenosis of 70% in a 2.0 mm vessel. The control coronary angiographies were performed after stent deployment at 12 +/- 2.8 months, and approximately 2 years of follow-up was completed. The polymer based paclitaxel-eluting stent has been shown to be effective in reducing restenosis. Patients were followed-up for 16.7 +/- 7.4 months. All patients survived after stent implantation, but 2 (1.3%) patients experienced acute myocardial infarction after 3 and 9 months following angioplasty. Recurrent angina pectoris was observed in 3 patients. Angiographic evidence of restenosis was observed in these 5 patients. Three patients underwent angioplasty because of re- stenosis, and coronary artery bypass grafting was conducted in the other 2 patients. The results indicate that Taxus stents can be implanted with a very high success rate and have encouraging long-term angiographic and clinical results. PMID- 16479036 TI - Reciprocal changes in 12-lead electrocardiography can predict left main coronary artery lesion in patients with acute myocardial infarction. AB - Acute left main coronary artery (LMCA) occlusion may result in acute myocardial infarction (AMI) or sudden death. ST elevation in the aVR and V1 leads is reported to be valuable in recognizing LMCA occlusion. Early recognition of electrocardiogram (ECG) changes, such as reciprocal ST depression in other leads, is helpful in averting this disaster. This study aimed to determine the reciprocal ST segment depression of 12-lead ECGs associated with acute LMCA occlusion. From January 2000 to December 2004, 61 patients who underwent emergency percutaneous coronary intervention in 3 hospitals due to AMI associated with LMCA (n = 18) and a left anterior descending coronary artery (LADCA) (n = 43) proximal lesion were selected. Reciprocal ST segment depression occurred in leads aVF, V(2), V(3), V(4), V(5), and V(6) with significantly higher incidence in the LMCA group than in the LADCA group. Stepwise linear multivariate discriminant analysis indicated that ST segment depression in leads aVF, V(2), and V(4) could distinguish the LMCA group from the LADCA group. We concluded that reciprocal ST segment depression in leads V(2), V(4), and aVF of a 12-lead ECG is an important predictor of acute LMCA occlusion. PMID- 16479037 TI - Bilateral cardiac catheterizations: the safety and feasibility of a superficial forearm venous and transradial arterial approach. AB - The transradial approach for left heart catheterization has become increasingly popular recently because of its clinical benefits. We examined the safety and feasibility of a transforearm approach for bilateral cardiac catheterizations, using the radial artery and a superficial forearm vein (the cephalic, basilic, or median antecubital vein). Between August 2002 and October 2003, 296 right heart catheterizations were performed in our hospital. A superficial forearm vein was used in one group of 101 patients, of which 98 had a concomitant left heart catheterization through the radial artery. The femoral vein was used for right heart catheterization in the second group of 195 patients. Of these patients, 37 underwent left heart catheterization through the radial artery and 157 through the femoral artery. All instances of bilateral catheterizations were successful except for one complication of pseudoaneurysm occurring in the transfemoral group. The procedure time for right heart catheterization was significantly less in the forearm group than the femoral group. The transforearm group had a larger proportion of males and of patients undergoing diagnostic right heart catheterization for congestive heart failure, dilated cardiomyopathy, and ischemic cardiomyopathy. Patients with aortic stenosis (AS), atrial septal defect (ASD), and mitral stenosis (MS) were mainly restricted to the transfemoral approach. We conclude that the transradial artery and superficial forearm venous approach for bilateral cardiac catheterizations is a safe and feasible alternative to the femoral approach in a wide range of patients, with the exception of patients with AS, ASD, or MS. PMID- 16479038 TI - Predictive factors of TIMI-3 flow before percutaneous coronary intervention in facilitated percutaneous coronary intervention for acute myocardial infarction. AB - The outcome for facilitated percutaneous coronary intervention (PCI) for acute myocardial infarction (AMI) is known to be more favorable in cases in which TIMI 3 flow is obtained before PCI. We investigated factors that affect the acquisition of TIMI-3 flow before PCI. Facilitated PCI was performed on 178 patients divided into two groups, a group in which TIMI-3 flow was acquired before conducting PCI and another in which it was not, and their background factors and short-term outcomes were investigated. The hemoglobin concentrations, white blood cell (WBC) counts, and HbA1c values were significantly lower in the group in which TIMI-3 flow was acquired before PCI and significantly more had a history of past smoking. According to the results of logistic analysis, WBC count (odds ratio [OR], 0.865, P = 0.0077), hemoglobin concentration (OR, 0.77, P = 0.0257), and smoking history (OR, 0.266, P = 0.0021) were independent factors that predicted acquisition of TIMI-3 flow. The WBC count and hemoglobin value on arrival at the emergency room and history of smoking were shown to be independent factors for acquisition of TIMI-3 flow before PCI in facilitated PCI. PMID- 16479039 TI - Impact of aspirin treatment on long-term outcome (over 10 years) after percutaneous coronary intervention. AB - Aspirin has been shown to reduce cardiovascular morbidity and mortality following percutaneous coronary intervention (PCI). However, its effects on long-term (over 10 years) mortality have not been fully elucidated. This retrospective study recorded the patient characteristics and admission medication for all patients undergoing PCI over an 8-year period from 1984 to 1992. Follow-up information was available for 748 patients (100%) for a mean of 143.6 +/- 43.4 months. A propensity analysis was performed to adjust for presumed selection biases in the administration of aspirin. The baseline clinical characteristics were similar between the group that received aspirin and the group that did not, except for the administration of statins and PCI procedural success rate. Of the 748 patients, 535 (71.5%) received aspirin treatment at the time of PCI. During the 12-year follow-up, 54 patients died from any cause and 20 patients from cardiac death. Kaplan-Meier analysis showed that aspirin treatment led to a significant reduction in all cause mortality (10% versus 16.4%; P = 0.01) and cardiac death (3.7% versus 8.0%; P = 0.02) compared to other antiplatelet drugs. The hazard ratio (HR) for the total mortality and cardiac mortality rates was adjusted using the Cox-proportional hazard model for confounding variables and propensity score. The all cause (HR, 0.49; 95%CI [0.29-0.80], P = 0.005) and cardiac mortality rates (HR, 0.32; 95%CI [0.14-0.72], P = 0.006) for patients receiving aspirin remained lower than for those not receiving aspirin. Aspirin treatment at the time of PCI significantly reduced the risk of death from any cause and cardiac death. The administration of aspirin had a positive impact on the over 10-year long-term outcomes of patients who underwent PCI. PMID- 16479040 TI - Long-term outcome after primary stenting versus balloon angioplasty for acute myocardial infarction. AB - The objective of the present prospective multicenter case-control study was to investigate the long-term clinical outcome (5 years) of primary stenting compared to primary percutaneous transluminal coronary angioplasty (PTCA) without stenting (POBA) in patients with acute myocardial infarction at 7 cardiovascular centers in Hokkaido, Japan. Forty-one patients with acute myocardial infarction treated with successful primary stenting (stent group: case) and paired with 41 matched control subjects with acute myocardial infarction treated by successful primary PTCA without stenting (POBA group: control) were analyzed. After 1 year, the stent group had a lower incidence of the combined clinical endpoint (death, rehospitalization due to congestive heart failure, nonfatal myocardial infarction, repeat angioplasty, CABG, or cerebrovascular events) compared to the POBA group (17.1% versus 39.0%, P = 0.049). After 5 years, the incidences of congestive heart failure and cardiac death were the same in both groups. However, compared to the POBA group, the stent group had a lower combined clinical endpoint (34.1% versus 61.0%, P = 0.027). The Kaplan-Meier event-free survival curves of the stent group showed a significantly lower occurrence of clinical events compared to the POBA group (P = 0.0116). Multiple logistic regression analysis of clinical events identified age > or = 69 years (P = 0.0092, odds ratio = 4.179) and stenting (P = 0.0158, odds ratio = 0.279) as explanatory factors. Compared with POBA, primary stenting for acute myocardial infarction results in a better long-term clinical outcome. PMID- 16479041 TI - Quality of life in the elderly after coronary bypass surgery. AB - In addition to the clinical outcome, a patient's perspective and satisfaction with their health status have become important indicators. One of the most common measures to assess the quality of life is Short Form (SF)-36. The objective of the present study was to measure the functional status of elderly patients who had undergone coronary bypass surgery and to evaluate the impact of that surgery on their quality of life. The study involved 120 nonsmoking patients who underwent coronary bypass surgery between January 1, 2001 and January 1, 2003 at the Sevket Demirel Heart Center. Assessments were made using physical, clinical, and laboratory findings. We used the Turkish version of the Short Form (SF)-36 preoperatively and 18 months after surgery. The paired t test, two-tailed correlation, and variant analysis were used for statistical analysis. Of the 120 patients, 108 could be followed during the study period. Significant physical and mental improvements were seen in all areas, especially in the items of vitality and mental health. Females seemed to benefit from surgery more than males. Cardiac surgery substantially improved the quality of life of our patients. The findings allowed us to determine the patient's perspective of his or her outcome. With this knowledge, health care workers can provide information to the patient regarding functional limitations after cardiac surgery. PMID- 16479042 TI - Pulmonary segmental artery ratio: an alternative to the pulmonary artery index in patients with tetralogy of fallot. AB - The objective of this study was to determine a reliable, alternative ratio to the pulmonary artery (PA) index, which will help to estimate the adequacy of postoperative pulmonary blood flow in patients with tetralogy of Fallot. We propose the pulmonary segmental artery ratio (PSAR), which is an angiographic measure for the quantitative standardization of the total number of pulmonary segmental arteries in a patient. The expected value of the PSAR is 1 and it is constant after the 16(th) week of intrauterine life. Retrospective analysis of the PSAR and PA index calculations in patients with tetralogy of Fallot was conducted. Sixty-one patients were assigned to a moderate or low risk group according to their PSAR; the low risk group included 31 patients whose PSAR was between 0.75-1 (group 1) while the moderate risk group included 30 patients whose PSAR was between 0.50-0.75 (group 2). High risk patients whose PSAR was less than 0.50 were excluded from the study. Postoperative peak right ventricular pressure, the pulmonary artery to systemic pressure ratio, and peripheral arterial oxygen saturation preoperatively after cardiopulmonary bypass were analyzed separately in groups 1 and 2. Postoperative peak right ventricular pressure was lower in group 1 than group 2, while the pulmonary artery to systemic pressure ratio and peripheral arterial oxygen saturation were higher in group 1 than group 2 (P < 0.01). Based on the present findings, it is concluded that PSAR is not as reliable as the Nakata index. However, in cases in which the PSAR and PA index are not correlated, PSAR may be helpful for determining the adequacy of postoperative pulmonary blood flow and postoperative outcomes of patients with hypoplastic pulmonary arteries. PMID- 16479043 TI - Impaired systemic endothelial function in patients with pseudoexfoliation syndrome. AB - Pseudoexfoliation syndrome (PEX) is the most common clinical precursor of open angle glaucoma. Recent studies have shown that pseudoexfoliative material is widely distributed throughout the body, including blood vessels. The aim of our study was to evaluate endothelial function in the brachial artery of patients with pseudoexfoliation syndrome. We prospectively examined 23 patients with PEX (mean age, 70 +/- 8 years) and 20 healthy age- and sex-matched individuals (mean age, 68 +/- 9 years) as a control group. Brachial artery endothelial function was assessed by vascular response to reactive hyperemia (flow-mediated dilation (FMD) and sublingual nitroglycerin (NTG-mediated dilation) using high-resolution ultrasound. Flow-mediated and NTG-induced dilation were expressed as the percent change in diameter after reactive hyperemia and after NTG administration relative to the baseline value, respectively. Patients with cardiovascular disease and other conditions associated with endothelial dysfunction were excluded. When compared with controls, patients with PEX had significantly lower flow-mediated dilation (4.5 +/- 2.8 versus 8.2 +/- 3.7, P = 0.01) and NTG-mediated dilation (10.9 +/- 3.1 versus 15.8 +/- 3.8, P = 0.0001). Flow-mediated dilation and NTG mediated dilation were similar in PEX patients with glaucoma (n = 11) and without glaucoma (n = 12). Flow-mediated and NTG-mediated dilation did not correlate with any measured parameter in any patient or control subject. The findings indicate that systemic endothelial function is impaired in PEX syndrome patients. PMID- 16479044 TI - Amiodarone and N-desethylamiodarone enhance endothelial nitric oxide production in human endothelial cells. AB - Amiodarone (AM) is a potent vasodilator and exhibits diverse cardiovascular protective effects in vivo, but their underlying mechanisms remain unsettled. We investigated the effects of AM and N-desethylamiodarone (DEA), the major metabolite of AM, on endothelial nitric oxide (NO) production using cultured human umbilical vein endothelial cells (HUVECs). The release of NO was evaluated as measured by nitrite, a stable metabolite of NO, using the Griess reaction and also measured directly by a NO-selective electrode. The expression of each nitric oxide synthase (NOS) mRNA was examined by reverse transcriptase-polymerase chain reaction (RT-PCR), and the effects of AM on eNOS mRNA expression were studied by quantitative real-time RT-PCR. AM and DEA (1-30 microM) enhanced NO production in a concentration-dependent manner. DEA was capable of producing more NO than AM. L NAME, a nonselective NOS inhibitor, EGTA, a Ca(2+)-chelating agent, and nickel, a nonspecific Ca(2+) blocker, all inhibited AM-induced NO production. However, LY294002, an Akt pathway inhibitor and SB202190, a MAP kinase inhibitor, did not significantly suppress the production. In RT-PCR analysis, only eNOS mRNA was detected. Treatment with AM for 4 hours did not show a significant increase in the expression of eNOS mRNA. AM lower than 30 microM did not induce apoptosis, net cell loss, or LDH release from cells. The present study provides the first evidence that therapeutic concentrations of AM and DEA enhance eNOS-mediated NO production without any toxic or apoptotic effects. This mechanism may underlie the cardiovascular protective effects of AM and its metabolite observed in a clinical setting. PMID- 16479045 TI - No age-related change in circulating endothelial progenitor cells in healthy subjects. AB - Endothelial progenitor cells were isolated from peripheral blood obtained from 32 healthy volunteers without cardiovascular risk factors who ranged in age from 20 to 61 years (mean [+/- SD] age, 34.1 +/- 9.6 years). The fractions of CD34(+) endothelial progenitor cells expressing kinase insert domain receptor-1, CD62E, or CD31 were analyzed with flow cytometry. Correlation analysis demonstrated that there was no significant correlation between subject age and the fraction of circulating CD34(+) mononuclear cells expressing kinase insert domain receptor-1 (P = 0.324; r = -0.180). Similarly, there was no significant correlation between subject age and the fraction of circulating CD34(+) mononuclear cells expressing CD62E (P = 0.496; r = -0.125) or the fraction of circulating CD34(+) mononuclear cells expressing CD31 (P = 0.245; r = -0.212). In conclusion, the experimental results showed that there was no age-related change in the basal level of circulating endothelial progenitor cells in healthy subjects without cardiovascular risk factors. PMID- 16479046 TI - Impaired expression of cardiac adiponectin in leptin-deficient mice with viral myocarditis. AB - A mouse model of encephalomyocarditis (EMC) virus-induced myocarditis was used to investigate the expression of adiponectin in damaged cardiomyocytes. We intraperitoneally injected EMC virus into leptin-deficient ob/ob (OB) mice and wild-type (WT) mice. OB mice were divided into two subgroups consisting of mice with no intervention and mice receiving leptin replacement starting simultaneously with viral inoculation. We determined differences in heart weight, cardiac histological score, numbers of infiltrating and apoptotic cells in the myocardium, expression levels of adiponectin and TNF-alpha mRNA in the heart, adiponectin immunoreactivity in myocytes, adiponectin and TNF-alpha concentrations in the heart, and immunoreactivity of adiponectin receptors in myocytes between OB mice and WT mice. There was significantly decreased adiponectin mRNA expression, immunoreactivity, and protein level in the heart, and reduced immunoreactivity of adiponectin receptor 1 in myocytes from OB mice on days 4 and 8 after viral inoculation as compared with those in WT mice, together with increased cardiac weight, severe inflammatory myocardial damage, and increased levels of cardiac TNF-alpha mRNA and protein. Replacement of leptin in OB mice inhibited the development of severe myocarditis through augmentation of adiponectin mRNA, immunoreactivity, and protein level, increased adiponectin receptor 1 immunoreactivity in myocytes, and suppressed levels of TNF-alpha mRNA and protein. These results suggest that impaired expression of cardiac adiponectin may contribute to the progression of viral myocarditis through enhanced expression of TNF-alpha under a leptin-deficient condition. PMID- 16479047 TI - Embolised stent into the circumflex coronary artery during percutaneous coronary intervention. AB - Dislodgement and embolisation of the new generation of coronary stents before deployment are rare. If it is impossible to withdraw the embolised stent from the coronary artery, the stent may be crushed into the side wall of the coronary artery with a balloon over a wire passed alongside the embolised stent. PMID- 16479048 TI - Pure ventricular septal myocardial infarction in a young man with coronary artery ectasia. AB - The case of a 30-year-old man with myocardial infarction localized in the interventricular septum is described. Coronary angiography performed on day 28 after the onset of symptoms revealed ectasia in the right and left coronary arteries, but no overt stenotic or occlusive lesions were present. Spasm was induced in the first septal branch of the left anterior descending artery by an acetylcholine provocation test, and single photon emission computed tomography myocardial perfusion imaging showed a reduced thallium-201 uptake localized in the interventricular septum. PMID- 16479049 TI - Facilitation of rotablator-stenting by measuring angiographic lesion length with the navicath (type-MUTO), a new measuring microcatheter. AB - In the present study, the usefulness and feasibility of a new measuring microcatheter, the Navicath (Type-MUTO) microcatheter, for facilitating the stenting procedure subsequent to ablation using a rotablator (rota-stenting) is described. A method for measuring the length of the targeted lesion (lesion length) angiographically with the Navicath when exchanging the guide wires is presented using 2 representative cases of rota-stenting. In addition, the validity of the selected stent according to the measurement of lesion length with the Navicath was evaluated by comparing the length of the selected stent with lesion length before PCI measured by quantitative coronary angiography. Based on the results obtained, we believe the Navicath is useful for facilitating rota stenting, and may even be applicable to PCI in general. PMID- 16479050 TI - Coronary artery fistula from the left circumflex to the coronary sinus. AB - A 27-year-old woman, who had received mitral valve repair for mitral regurgitation resulting from infective endocarditis, was admitted for a close examination of abnormal echocardiographic findings in the left atrium. Transthoracic echocardiography showed trivial mitral regurgitation with normal left ventricular contraction and dilatation of the coronary sinus. Auscultation revealed a grade 2 continuous murmur along the left sternal border. Transesophageal echocardiography demonstrated a marked dilatation of the coronary sinus just behind the posterior wall of the left atrium and turbulent blood flow in the dilated coronary sinus. Cardiac catheterization showed no significant step up of oxygen saturation in the right heart and normal pulmonary artery pressure. Coronary angiography revealed a markedly dilated and tortuous circumflex coronary artery connected to the coronary sinus through a fistula. A left circumflex artery with a fistulous connection to the coronary sinus is extremely rare. PMID- 16479051 TI - Variation in parasystolic cycle length. AB - At the time of the first visit to our clinic, an electrocardiographic examination of a 73-year-old female patient revealed ventricular premature contractions (VPCs) with variable coupling intervals that were diagnosed as parasystole. Characteristically many of the parasystoles had no sinus contractions between two consecutive VPCs, which we referred to as pure parasystole. We first repeatedly examined variations in the length of the parasystolic cycles between January 6, 1997 and March 2, 2003 using electrocardiography. The time courses recorded over this period showed that the length of the parasystolic cycle did not remain constant, but varied irregularly within a relatively narrow range. We also recorded the length of the parasystolic cycles over 3 hours using Holter monitoring. The interectopic intervals plotted against mean sinus cycle length showed that the cycle length of pure parasystoles remained almost constant at about 1,300 ms over the 3 hours. We also examined the cycle length during exercise and found that it was slightly prolonged thereafter, while the sinus cycle length was clearly shortened after exercise. The average of six deep breathing tests showed that parasystolic cycle length did not significantly differ between deep inspiration and deep expiration, whereas the sinus cycle length during expiration was significantly longer than that during inspiration. These results indicate that the responses to both exercise and deep breathing obviously differed between the parasystolic and sinus cycle lengths. PMID- 16479052 TI - Lossless compression based on inter-frame coding for MDCT. AB - Many images are generated by medical equipment and the number of images to be stored increases yearly. To cope with the large digital storage and transmission requirements, data compression techniques have been studied. In the present study, grayscale data (2 bytes/pixel) were divided into high bytes and low bytes, and then the original medical images were also divided into high byte (1byte/pixel format) and low byte (1byte/pixel format) images. Inter-frame coding with and without division into high and low byte images was applied to MDCT images. Prediction (inter-frame) coding is a very common method to compress sequence images. The division process can be added to inter-frame coding easily. The compression ratio of the archived file produced by application of Lemple-Ziv compression and prediction coding with the division process was 0.39 at 700 images. PMID- 16479053 TI - [A Generator of Mono-energetic Electrons for Response Test of Charged Particle Detectors.]. AB - We designed and fabricated a generator of mono-energetic electrons for the response test of charged particle detectors, which is used to measure fragmented particles of the carbon beam for cancer therapy. Mono-energetic electrons are extracted from (90)Sr by analyzing the energy of beta rays in the generator with a magnetic field. We evaluated performance parameters of the generator such as the absolute energy, the energy resolution and the counting rates of extracted electrons. The generator supplies mono-energetic electrons from 0.5MeV to 1.7MeV with the energy resolution of 20% in FWHM at higher energies than 1.0MeV. The counting rate of electrons is 400cpm at the maximum when the activity of (90)Sr is 298kBq. The generator was used to measure responses of fragmented-particle detectors and to determine the threshold energy of the detectors. We evaluated the dependence of pulse height variation on the detector position and the threshold energy by using the generator. We concluded this generator is useful for the response test of general charged particle detectors. PMID- 16479054 TI - [Sensitometry of Mammographic Screen-film System Using Bootstrap Aluminum Step Wedge.]. AB - Recently, a few types of step-wedges for bootstrap sensitometry with a mammographic screen-film system have been proposed. In this study, the bootstrap sensitometry with the mammographic screen-film system was studied for two types of aluminum step-wedges. Characteristic X-ray energy curves were determined using mammographic and general radiographic aluminum step-wedges devised to prevent scattered X-rays generated from one step penetrating into the region of another one, and dependence of the characteristic curves on the wedges was also discussed. No difference was found in the characteristic curves due to the difference in the step-wedges for mammography and general radiography although there was a slight difference in shape at the shoulder portion for the two types of step-wedges. Therefore, it was concluded that aluminum step-wedges for mammography and general radiography could be employed in bootstrap sensitometry with the mammographic screen-film system. PMID- 16479055 TI - Increased expression of sodium transporters in rats chronically inhibited of nitric oxide synthesis. AB - The present study was done to determine whether endogenous nitric oxide (NO) plays a role in the regulation of sodium transporters in the kidney. Male Sprague Dawley rats were treated with NG-nitro-L-arginine methyl ester (L-NAME, 100 mg/L drinking water) for 4 weeks. Control rats were supplied with tap water without drugs. Expression of Na, K-ATPase, type 3 Na/H exchanger (NHE3), Na/K/2Cl cotransporter (BSC1), and thiazide-sensitive Na/Cl cotransporter (TSC) proteins was determined in the kidney by Western blot analysis. Catalytic activity of Na,K ATPase was also determined. The treatment with L-NAME significantly and steadily increased the systemic blood pressure. Total and fractional excretion of urinary sodium decreased significantly, while creatinine clearance remained unaltered. Neither plasma renin activity nor aldosterone concentration was significantly altered. The alpha1 subunit expression and the catalytic activity of Na, K-ATPase were increased in the kidney. The expression of NHE3, BSC1 and TSC was also increased significantly. These results suggest that endogenously-derived NO exerts a tonic inhibitory effect on the expression of sodium transporters, including Na, K-ATPase, NHE3, BSC1, and TSC, in the kidney. PMID- 16479056 TI - Transfusion-related cytomegalovirus infection among very low birth weight infants in an endemic area. AB - This study investigated the incidence of acquired cytomegalovirus (CMV) infection in very low birth weight infants (VLBWI) given CMV seropositive blood, and sought to determine whether filtering and irradiation of blood products could help prevent CMV infection and the time required to clear passively-derived anti-CMV IgG among 80 VLBWI transfused with filtered-irradiated blood, 20 VLBWI transfused with nonfiltered- nonirradiated blood and 26 nontransfused VLBWI. CMV IgG and IgM values were obtained from all blood products prior to transfusions, and from VLBWI at birth until the infants became seronegative. Urine was obtained for CMV culture at birth and every 3-4 weeks until 12 weeks after the final transfusion. The incidence of CMV IgG seropositivity among the 126 infants at birth and the blood products given were 96% and 95%, respectively. The incidence of acquired CMV infection was 4/100 (4%) in the transfused group: 2/80 (2.5%) and 2/20 (10%) in the filtered-irradiated and nonfiltered-nonirradiated transfusion groups, respectively. Approximately 9-10 months elapsed to clear passively acquired CMV IgG. The irradiation and filtering of the blood products did not seem to decrease the transfusion-related CMV infection rate in Korea among VLBWI, however, further validation is recommended in a larger cohort of infants. PMID- 16479057 TI - Threshold of nuchal translucency for the detection of chromosomal aberration: comparison of different cut-offs. AB - This study evaluated the sensitivities and false positive rates of the screening test using ultrasonographic measurement of thickness of nuchal translucency (NT) with different cut-offs for chromosomal aberration in a Korean population. We included 2,570 singleton pregnancies undergoing ultrasound between 11 weeks and 14 weeks of gestation in this study. We analyzed the sensitivities of NT alone for screening chromosomal aberration using three cut-offs -2.5 mm, 3.0 mm, and 95th percentile for each crown rump length (CRL). There were 31 chromosomal aberrations (1.2%) including 12 cases of trisomy 21. The numbers of chromosomal aberrations that were detected by NT with different cut-offs of 2.5 mm, 3.0 mm and the 95th percentile CRL were 22, 18 and 23, respectively. At a threshold of 2.5 mm, the sensitivity and the false positive rate for total chromosomal aberrations were 67.7% and 6.3%, respectively. At 3.0 mm, those were 54.8% and 3.5%, respectively. At the 95th percentile CRL, those were 70.9% and 5.8%, respectively. The use of CRL-dependent cut-offs for nuchal translucency improves the detection of chromosomal aberrations when compared to fixed cut-offs in a Korean population. PMID- 16479058 TI - Risk factor analysis for development of asymptomatic carotid stenosis in Koreans. AB - Many risk factors for atherosclerosis have been proposed to identify high risk individuals. We conducted a retrospective study to determine the risk factors for development of carotid stenosis (CS) in Koreans. Database of 2,805 subjects who underwent a check up of carotid artery for health examination were analyzed. Stenosis (%) of common carotid artery or proximal internal carotid artery was examined with ultrasonography. Subjects were divided into 2 groups (Group I; CS <10%, Group II; CS >or= 30%). We compared demographic, laboratory and clinical data between 2 groups to determine the risk factors of CS. One hundred ninety seven subjects (7.0%) were categorized as Group II. At age- and sex-adjusted multivariate analysis, diabetes mellitus, hypertension, cerebrovascular disease, ischemic heart disease, hyperlipidemia, aspirin medication, current smoking, fasting glucose, total cholesterol, low density lipoprotein-cholesterol (LDL-C) and leukocyte count were significant risk factors of CS. At stepwise logistic regression analysis, age, hypertension, hyperlipidemia, LDL-C and leukocyte count were independent risk factors. At subgroup analysis by smoking, age and leukocyte count were independent risk factors in smoker and age and hypertension in nonsmoker. PMID- 16479059 TI - Surgical management of Takayasu's arteritis. AB - The results of surgical bypass and endarterectomy in Takayasu's arteritis (TA) were reported to be poor compared to usual atherosclerosis patients. However, if ischemic symptoms due to occlusive disease were severe, surgical procedures were inevitable. We report surgical experience of 5 patients with TA. Five women (ranged from 26 to 58 yr) were operated between June 1998 and May 2004. Three patients showed occlusion of main branches of aortic arch and had symptoms of cerebral ischemia. One patient showed near total occlusion in the midabdominal aorta and had symptoms of orthopnea and uncontrolled hypertension. One patient showed total occlusion of abdominal aorta at the level of aortic bifurcation and had a symptom of severe claudication on both legs. Bypasses from the ascending aorta to the carotid artery were performed in 3 cases. Bypass from the thoracic aorta to the left common iliac artery was performed in one case and endarterectomy of abdominal aorta in one case. The ischemic symptoms related with arterial occlusion were resolved after surgery. And the symptoms of cardiac failure disappeared. The symptomatic TA frequently required arterial reconstruction. The symptomatic improvement and excellent mid-term patency could be expected after arterial reconstruction and endarterectomy. PMID- 16479060 TI - Aprotinin attenuates the elevation of pulmonary vascular resistance after cardiopulmonary bypass. AB - Pulmonary vascular resistance (PVR) is generally believed to be elevated after cardiopulmonary bypass (CPB) due to whole body inflammation. Aprotinin has an antiinflammatory action, and it was hypothesized that aprotinin would attenuate the PVR increase induced by CPB. Ten mongrel dogs were placed under moderately hypothermic CPB for 2 hr. The experimental animals were divided into a control group (n=5, group I) and an aprotinin group (n=5, group II). In group II, aprotinin was administered during pre-bypass (50,000 KIU/kg) and post-bypass (10,000 KIU/kg) periods. Additional aprotinin (50,000 KIU/kg) was mixed in CPB priming solution. PVRs at pre-bypass and post-bypass 0, 1, 2, 3 hr were calculated, and lung tissue was obtained after the experiment. Post-bypass PVRs were significantly higher than prebypass levels in all animals (n=10, p<0.001). PVR elevation in group II was less than in group I at 3 hr post-bypass (p=0.0047). Water content of the lung was lower in group II (74+/-9.4%) compared to that of group I (83+/-9.5%), but the difference did not reach significance (p=0.076). Pathological examination showed a near normal lung structure in group II, whereas various inflammatory reactions were observed in group I. We concluded that aprotinin may attenuate CPB-induced PVR elevation through its anti inflammatory effect. PMID- 16479061 TI - Impact of circulating TGF-Beta and IL-10 on T cell cytokines in patients with asthma and tuberculosis. AB - Regulatory T cells, which stimulate or inhibit the effector functions of distinct T cell subsets, are critical in the control of the immune response. We investigated the effect of TGF-beta and IL-10 on T cell subsets according to the Th1/Th2 immune status. Sixty-two patients with asthma and 38 patients with pulmonary tuberculosis were included. Allergy skin tests, tuberculin tests, and chest radiography were performed. The levels of circulating IL-4, IFN-gamma, TGF beta1, and IL-10 were measured using ELISA. The level of TGF-beta1 was higher in patients with asthma than in those with tuberculosis, but the IL-10 levels were the same between the asthma and tuberculosis groups. Atopy was unrelated to the tuberculin response. The IFN-gamma level was correlated with the IL-10 level, and the level of IL-4 was unrelated to the IL-10 or TGF-beta1 level. The level of IL 10 was higher in the negative tuberculin reactors than in the positive tuberculin reactors among patients with asthma, and TGF-beta1 was higher in the positive tuberculin reactors than in the negative tuberculin reactors among patients with tuberculosis. These results demonstrate that the regulatory effects of circulating TGF-beta and IL-10 on T cell cytokines may be different between Th2 type asthma and Th1 tuberculosis. PMID- 16479062 TI - Prognostic value of immunohistochemical staining of p53, bcl-2, and Ki-67 in small cell lung cancer. AB - Small cell lung cancer (SCLC) is one of the most fatal cancers in humans and many factors are known to be related to its poor prognosis. Immunohistochemical (IHC) stainings were done on SCLC specimens in order to investigate the prognostic value of the apoptosis-related gene expression and the tumor proliferative maker, and the relationships among these IHC results and patients clinical characteristics, chemoresponsiveness, and survival were analyzed. The medical records of 107 patients were reviewed retrospectively. IHC stainings for p53, bcl 2 and Ki-67 expressions were performed in the 66 paraffin-embedded biopsy samples. Sixty-six out of the 107 patients were evaluable for response rate and survival. The overall response rate was 75% (95% Confidence Interval=74-76%) and the median survival time was 14 months. The median survival time of limited stage was 16 months and that of extensive stage was 10 months. The prevalence of p53, bcl-2 and Ki-67 expression was 62%, 70%, and 49%, respectively. There were no correlations among the immunoreactivities of p53, bcl-2 and Ki-67 with clinical stage, chemoresponsiveness or overall survival. The clinical stage was the only prognostic factor influencing survival. The expression rates of p53, bcl-2, and Ki-67 were relatively high in SCLC without any prognostic significance. The exact clinical role of these markers should be defined through further investigations. PMID- 16479064 TI - Clinical significance of platelet count at day +60 after allogeneic peripheral blood stem cell transplantation. AB - Thrombocytopenia (TP) is a frequent complication after allogeneic stem cell transplantation (SCT) and regarded as a poor prognostic factor, especially in patients with chronic graft-versus-host disease (GVHD), although various factors were related to the development of TP after allogeneic SCT. Sixty-three patients receiving allogeneic peripheral blood stem cell transplantation (PBSCT) were stratified according to platelet count (PC) at day +60 and analyzed in terms of overall survival (OS) and the incidence of non-relapse mortality (NRM). Ten patients (15.9%) were stratified in group 1 (PC /= 80 x 10(9)/L). Group 3 was associated with lower incidence of extensive chronic GVHD (p=0.013), better 3-yr OS (p=0.0030), and lower NRM rate (p<0.0001). In multivariate analyses, the PC at day +60 was identified as an independent prognostic factor (p=0.003) together with CD34+ cell dose (p<0.001), disease risk (p=0.004), and acute GVHD (p=0.033) in terms of NRM, and the PC (p=0.047) and CD34+ cell dose (p=0.026) in terms of incidence of infectious events. Measuring the platelet count at day +60 is a simple method for predicting the risk of chronic GVHD development and prognosis after allogeneic PBSCT. PMID- 16479065 TI - Comparison of the efficacy of oral capecitabine versus bolus 5-FU in preoperative radiotherapy of locally advanced rectal cancer. AB - The effects of treatment with oral capecitabine vs. bolus 5-FU, administered concurrently with preoperative radiotherapy, were compared in the treatment of locally advanced rectal cancer (LARC). One hundred and twenty-seven patients with LARC received concurrent preoperative chemoradiation using two cycles bolus 5-FU (500 mg/m2/day) plus leucovorin (LV, 20 mg/m2/day) (Group I). Another LARC group received concurrent chemoradiation using two cycles 1,650 mg/m2/day of oral capecitabine and 20 mg/m2/day of LV (Group II, 97 patients). Radiation was delivered to the primary tumor at 50.4 Gy in both groups. Definitive surgery was performed 6 weeks after the completion of chemoradiation. A pathologic complete remission was achieved in 11.4% of patients in Group I and in 22.2% of patients in Group II (p= 0.042). The down-staging rates of the primary tumor and lymph nodes were 39.0/ 68.7% in Group I and 61.1/87.5% in Group II (p=0.002/0.005). Sphincter-preserving surgery was possible in 42.1% of patients in Group I and 66.7% of those in Group II (p=0.021). Grade 3 or 4 leucopenia, diarrhea, and radiation dermatitis were statistically more prevalent in Group I than in Group II, while the opposite was true for grade 3 hand-foot syndrome. Preoperative chemoradiation using oral capecitabine was better tolerated than bolus 5-FU and was more effective in the promotion of both down-staging and sphincter preservation in patients with LARC. PMID- 16479063 TI - The hematologic response to anti-apoptotic cytokine therapy: results of pentoxifylline, ciprofloxacin, and dexamethasone treatment for patients with myelodysplastic syndrome. AB - TNF-alpha mediated apoptosis of the hematopoietic cells has been thought to contribute to the ineffective hematopoiesis observed in myelodysplastic syndrome (MDS). The combination of pentoxifylline (P) and ciprofloxacin (C) has been shown to reduce the serum levels of TNF-alpha, and an earlier trial of P and C with dexamethasone (D) provided good palliation for patients with MDS. The purpose of this study is to assess the hematologic response to PCD therapy for patients suffering with MDS. 21 of 25 patients who completed at least of 12 weeks of treatment were evaluable for the treatment efficacy. At baseline, the patient's median age was 60 yr (range: 18-75 yr). The diagnoses according to WHO classification included: RA (n=5), RCMD (n=10), RARS (n=1), RCMD/RS (n=1), RAEB (3), and CMML (n=1). 11 patients (52%) had at least single lineage response. 3 patients (11%) showed improvement of triple lineage cytopenia. There were no differences in the response rates between the FAB subtypes. The median time to response was 4 weeks (range: 2-12 weeks), and it is interesting that 9 of 11 patients who had a response remained without relapse for a median of 177 days (range: 78-634 days). These preliminary results indicate that anti-cytokine therapy with PCD is an effective and well tolerated palliative treatment for patients with MDS. PMID- 16479066 TI - Seroprevalence of hepatitis B virus among health care workers in Korea. AB - We studied the seroprevalence of HBsAg, anti-HBs and anti-HBc and the vaccination histories among health care workers (HCWs) at a large suburban referral hospital in Korea. The purpose of this study was to determine the immune status of HCWs against hepatitis B virus and we also wanted to prepare a practical guideline to protect HCWs from occupational exposure. During December, 2003, 571 HCWs (56 physicians, 289 nurses, 113 technicians and 113 aid-nurses) aged between 21 and 74 yr were included in the surveillance. The positive rates of HBsAg and anti-HBs were 2.4% (14/571) and 76.9% (439/571), respectively. The positive rate of anti HBs was lower in the physician group, and this was associated with the male gender and older age. Of the 439 anti-HBs positive cases, 320 cases (73.1%) were anti-HBc negative and this was significantly associated with a past history of HBV vaccination. The distribution of the anti-HBs levels was not associated with age (except for HCWs in their sixties), gender or occupation. Our study revealed that the seroprevalence rates of HBsAg and anti-HBs in HCWs in Korea were not different from those of the general population. Based on this surveillance, we can make reasonable decisions in case of occupational exposure to hepatitis B virus. PMID- 16479067 TI - Mesenchymal hamartomas of the liver: comparison of clinicopathologic features between cystic and solid forms. AB - Mesenchymal hamartoma (MH) of the liver is an uncommon benign lesion related to ductal plate malformation. It is usually cystic and mainly composed of myxoid mesenchymal tissue with tortuous or cystic bile ducts. In order to characterize the clinicopathological features of MH, the Korean Gastrointestinal Pathology Study Group collected a total of 17 MH cases diagnosed in 7 hospitals from 1992 to 2002 and compared the clinicopathologic findings of cystic MH with those of solid variant. Among the 17 cases, 7 (41%) were solid. The solid form showed a higher serum level of alpha-fetoprotein (AFP), the smaller bile ducts, and more frequent proliferation of vessels. Serum AFP level was related to the amount of hepatocytes. Two of seven solid cases harbored a larger amount of evenly distributed hepatocytes and proliferation of small duct with focal hepatocyte bile duct transition. These histologic findings are similar to those of mixed hamartoma. Therefore, the mixed hamartoma and the MH of both solid and cystic types could be the variants of one disease spectrum. And hepatocytes within MH might be rather a genuine tumor component than entrapped into the tumor. In conclusion, MH can show various clinicopathological features and recognition of these features will facilitate accurate diagnosis of MH. PMID- 16479068 TI - Initial experiences with laparoscopy-assisted and total laparoscopy for anatomical liver resection: a preliminary study. AB - Although laparoscopic surgery has become more popular, its technical difficulties have limited the applications of this technique to liver surgery. We report here on our experience with liver resection with using the laparoscopy-assisted (Lap Assist) and total laparoscopic (Total-Lap) methods. From April 2001 to June 2003, a total of 20 laparoscopic anatomical resections of the liver were retrospectively reviewed. These were comprised of 10 cases in which the Lap Assist method was used (these were performed during the early study period), and 10 cases in which the Total-Lap was used (these were done in the later study period). In the Lap-Assist group, the following resections were performed: 7 cases of left lateral sectionectomy, a case of left hemihepatectomy, a case of right hemihepatectomy and a case of open conversion. In the Total-Lap group, 6 cases of left hemihepatectomy and 4 cases of left lateral sectionectomy were performed. The sizes of the incisions were 8.7 cm and 4.6 cm, respectively, (p=0.000). There were no differences in the operation times, the transfusion amounts, the starting days of the patients' diets, the complication rates or the durations of the hospital stay between the two groups. Both the laparoscopy assisted method and the total laparoscopic method are feasible to use for performing anatomical liver resection. PMID- 16479069 TI - Assessment of fluid shifts of body compartments using both bioimpedance analysis and blood volume monitoring. AB - Fluid shifts are commonplace in chronic hemodialysis patients during the intra- and interdialytic periods. In this study, we evaluated fluid shifts of body compartments using both bioimpedance spectroscopy and blood volume monitoring from the start to the end of hemodialysis. 24 stable hemodialysis patients were included on the study. Relative change of blood volume was progressively reduced from the start to the end of hemodialysis (1 hr, -7.22+/-3.23%; 2 hr, -9.78+/ 4.69%; 3 hr, -12.88+/-5.65%; 4 hr, -15.41+/-6.54%, respectively). Mean % reduction of intracellular fluid was not significantly different to that of extracellular fluid at the end of hemodialysis (delta ICF, -6.58+/-5.34% vs. delta ECF, -7.07+/-5.12%). Mean % fluid reduction of arms, legs and trunk was 11.98+/-6.76%, -6.43+/-4.37% and -7.47+/-4.56%, respectively at the end of hemodialysis. There were 3 characteristic patterns in blood-volume change. Similar amounts of fluid were removed from the extracellular and intracellular compartments during hemodialysis, with the arms showing the greatest loss in terms of body segments. The pattern of blood volume change measured by blood volume monitoring may be useful for more accurate determination of dry-weight and for correcting volume status in hemodialysis patients. PMID- 16479070 TI - Expression and mutational analysis of c-kit in ovarian surface epithelial tumors. AB - Coexpression of Kit ligand and c-kit has been reported in some gynecologic tumors. To determine whether imatinib mesylate is useful in ovarian epithelial tumors, we performed immunohistochemical and mutational analysis. The cases consisted of 33 cases, which included 13 serous cystadenocarcinomas, 1 borderline serous tumor, 8 mucinous cystadenocarcinomas, 6 borderline mucinous tumors and 5 clear cell carcinomas. Five cases of serous cystadenoma and 5 cases of mucinous cystadenoma were also included. In the immunohistochemical study, 3 cases (3/6, 50%) of borderline mucinous cystic tumor and two cases (2/8, 25%) of mucinous cystadenocarcinoma show positive staining for KIT protein. Only one case (1/13, 7.7%) of serous cystadenocarcinoma had positive staining. On mutational analysis, no mutation was identified at exon 11. However, two cases of borderline mucinous tumors and one case of mucinous cystadenocarcinoma had mutations at exon 17. In these cases, the immunohistochemistry also shows focal positive staining at epithelial component. Although, KIT protein expression showed higher incidence in mucinous tumors than serous tumors, they lack KIT-activating mutations in exon 11. Thus, ovarian surface epithelial tumors are unlikely to respond to imatinib mesylate. PMID- 16479071 TI - Predictors of success of repeated injections of single-dose methotrexate regimen for tubal ectopic pregnancy. AB - The purpose of this study is to evaluate predictors of success of repeated injections of methotrexate in the single-dose regimen for the treatment of tubal ectopic pregnancy. All patients who had ectopic tubal pregnancy and were treated with a single dose regimen were retrospectively identified. 126 patients were treated with methotrexate. Among them, 39 patients were adequate for this study. 33 were treated with the 2nd dose and 27 were successfully cured. Additionally, 6 who were injected with the 3rd dose were all cured as well. Therefore, in our study, the success rate for the repeated injections of methotrexate was found to be 84.6% (33/39). The mean initial beta-hCG level was significantly lower in patients who were successfully treated than in patients who failed (3915.3+/ 3281.3 vs. 8379.7+/-2604.4 IU/mL, p<0.05). The success rate is 96% when the beta hCG level is less than 6,000 IU/mL and is 58% when beta-hCG is greater than 6,000 IU/mL (OR=18.57, 95% CI 1.86-185.89). The initial beta-hCG level is the only factor that has significant meaning as predictor of success of repeated injections of methotrexate in the single-dose regimen. Repeated injections of methotrexate may be particularly effective when the initial beta-hCG level is below 6,000 IU/mL. PMID- 16479072 TI - Analysis of the monocyte chemoattractant protein 1 -2518 promoter polymorphism in Korean patients with alopecia areata. AB - Monocyte chemoattractant protein-1 (MCP-1) levels are increased in scalp lesions of patients with alopecia areata (AA), suggesting a role in the development of AA. Recently, a biallelic A/G polymorphism in the MCP-1 promoter at position 2518 has been found, influencing the level of MCP-1 expression in response to an inflammatory stimulus. We investigated whether the presence of these polymorphisms were associated with AA in Korean population. 145 Korean patients with AA, 246 healthy subjects without clinical evidence of AA were screened for genotype with a PCR-based assay. In the AA patients the frequency of the A and G alleles was 40.3 and 59.7%, respectively and the distribution of the A/A, A/G and G/G genotypes was 19.3, 42.1 and 38.6%, respectively. Amongst the controls the frequency of the A and G alleles was 39.8 and 60.2%, and the distribution of the A/A, A/G, G/G genotypes in the same group was 17.5, 44.7 and 37.8%, respectively. There was no significant difference in the allele frequencies and genotype distributions between the patients and the controls (p=0.889, p=0.848, respectively). Our data indicates that no association exists between the -2518A/G polymorphism of the MCP-1 gene and susceptibility to alopecia areata. PMID- 16479073 TI - Ceramides and cell signaling molecules in psoriatic epidermis: reduced levels of ceramides, PKC-alpha, and JNK. AB - Ceramides are the main lipids in the stratum corneum and are generated during cellular stress and apoptosis by de novo synthesis or by the action of sphingomyelinase. In addition, they are lipid second messengers produced by sphingolipid metabolism and trigger important cell responses, including protein kinase C-alpha (PKC-alpha) activation and the stimulation of signal transduction pathways with apoptosis and stress-activated protein kinases (SAPK), such as c jun N-terminal kinase (JNK). Thus, ceramides have anti-proliferative and apoptotic effects. This study measured the changes in the levels of epidermal ceramides and ceramide-related apoptotic signaling molecules in psoriasis patients. Samples from lesional and non-lesional epidermis were obtained from psoriasis patients. Total ceramides were fractionated using thin-layer chromatography, and the levels of PKC-alpha and JNK expression were measured using Western blot analysis with specific antibodies. The ceramide level was reduced significantly, and this was associated with the downregulation of apoptotic signaling molecules, such as PKC-alpha and JNK, in the lesional epidermis of psoriasis patients. These results suggest that the decreased level of ceramides downregulates the apoptotic pathway, leading to epidermal proliferation in psoriasis. PMID- 16479074 TI - Evidence for cyclooxygenase-2 association with caveolin-3 in primary cultured rat chondrocytes. AB - The purpose of this study was to demonstrate the cellular localization of cyclooxygenase-2 (COX-2) and caveolin-3 (Cav-3) in primarily cultured rat chondrocytes. In normal rat chondrocytes, we observed relatively high levels of Cav-3 and a very low level of COX-2 mRNA and protein. Upon treating the chondrocytes with 5 microM of CdCl(2) (Cd) for 6 hr, the expressions of COX-2 mRNA and protein were increased with the decreased Cav-3 mRNA and protein expressions. The detergent insoluble caveolae-rich membranous fractions that were isolated from the rat chondrocytes and treated with Cd contained the both proteins of both COX-2 and Cav-3 in a same fraction. The immuno-precipitation experiments showed complex formation between the COX-2 and Cav-3 in the rat chondrocytes. Purified COX-2 with glutathione S-transferase-fused COX-2 also showed complex formation with Cav-3. Confocal and electron microscopy also demonstrated the co-localization of COX-2 and Cav-3 in the plasma membrane. The results from our current study show that COX-2 and Cav-3 are co-localized in the caveolae of the plasma membrane, and they form a protein-protein complex. The co localization of COX-2 with Cav-3 in the caveolae suggests that the caveolins might play an important role for regulating the function of COX-2. PMID- 16479076 TI - Influence of lamina terminalis fenestration on the occurrence of the shunt dependent hydrocephalus in anterior communicating artery aneurysmal subarachnoid hemorrhage. AB - Recently, it was reported that fenestration of the lamina terminalis (LT) may reduce the incidence of shunt-dependent hydrocephalus in aneurysmal subarachnoid hemorrhage (SAH). The authors investigated the efficacy of the LT opening on the incidence of shunt-dependent hydrocephalus in the ruptured anterior communicating artery (ACoA) aneurysms. The data of 71-ruptured ACoA aneurysm patients who underwent aneurysmal clipping in acute stage were reviewed retrospectively. Group I (n=36) included the patients with microsurgical fenestration of LT during surgery, Group II (n=35) consisted of patients in whom fenestration of LT was not feasible. The rate of shunt-dependent hydrocephalus was compared between two groups by logistic regression to control for confounding factors. Ventriculo peritoneal shunts were performed after aneurysmal obliteration in 18 patients (25.4%). The conversion rates from acute hydrocephalus on admission to chronic hydrocephalus in each group were 29.6% (Group I) and 58.8% (Group II), respectively. However, there was no significant correlation between the microsurgical fenestration and the rate of occurrence of shunt-dependent hydrocephalus (p>0.05). Surgeons should carefully decide the concomitant use of LT fenestration during surgery for the ruptured ACoA aneurysms because of the microsurgical fenestration of LT can play a negative role in reducing the incidence of chronic hydrocephalus. PMID- 16479077 TI - Radiofrequency neurotomy of cervical medial branches for chronic cervicobrachialgia. AB - Chronic neck and arm pain or cervicobrachialgia commonly occurs with the degeneration of cervical spine. Authors investigated the usefulness of radiofrequency (RF) neurotomies of cervical medial branches in patients with cervicobrachialgia and analyzed the factors which can influence the treatment outcome. Demographic data, types of pain distribution, responses of double controlled blocks, electrical stimulation parameters, numbers and levels of neurotomies, and surgical outcomes were evaluated after mean follow-up of 12 months. Pain distribution pattern was not significantly correlated with the results of diagnostic blocks. Average stimulation intensity was 0.45 V, ranging from 0.3 to 0.69, to elicit pain response in cervical medial branches. The most common involvement of nerve branches was C4 (89%), followed by C5 (82%), C6 (75%), and C7 (43%). Among total of 28 patients, nineteen (68%) reported successful outcome according to outcome criteria after 6 months of followup (p=0.001), and eight (42%) of 19 patients reported complete relief (100%) of pain. Four patients showed recurrence of pain between 6 and 12 months. It was therefore concluded that cervical medial branch neurotomy is considered useful therapeutic modality for the management of cervicobrachialgia in selected patients, particularly in degenerative zygapophyseal disorders. PMID- 16479075 TI - The spectrum of NF1 mutations in Korean patients with neurofibromatosis type 1. AB - Neurofibromatosis type 1 (NF1) is one of the most common autosomal dominant disorders in humans. NF1 is caused by mutations in the NF1 gene which consists of 57 exons and encodes a GTPase activating protein (GAP), neurofibromin. To date, more than 640 different NF1 mutations have been identified and registered in the Human Gene Mutation Database (HGMD). In order to assess the NF1 mutational spectrum in Korean NF1 patients, we screened 23 unrelated Korean NF1 patients for mutations in the coding region and splice sites of the NF1 gene. We have identified 21 distinct NF1 mutations in 22 patients. The mutations included 10 single base substitutions (3 missense and 7 nonsense), 10 splice site mutations, and 1 single base deletion. Eight mutations have been previously identified and thirteen mutations were novel. The mutations are evenly distributed across exon 3 through intron 47 of the NF1 gene and no mutational hot spots were found. This analysis revealed a wide spectrum of NF1 mutations in Korean patients. A genotype phenotype correlation analysis suggests that there is no clear relationship between specific NF1 mutations and clinical features of the disease. PMID- 16479078 TI - Prognostic implication of telomerase activity in patients with brain tumors. AB - Telomerase adds telomeric repeats to the ends of telomeres to compensate for their progressive loss. A favorable prognosis is associated with low or no telomerase in some tumors. The authors investigated whether telomerase activity is associated with survival of patients with brain tumors. Sixty-two consecutive patients with brain tumors underwent surgery, and their surgical specimens were investigated. The patients were pathologically categorized as group I (aggressive group) and group II (non-aggressive group). Telomerase activity was examined by the telomeric repeat amplification protocol (TRAP) assay. The median time was calculated in association with overall survival and progression-free survival in each group. The significant difference was noted in telomerase activity between high-grade gliomas and lowgrade gliomas (p=0.022). Telomerase activity was significantly associated with the median overall survival and progression-free survival in all tumors of the aggressive group. On the other hand, the median overall survival in the non-aggressive group was not dependent on telomerase activity, while the median progression-free survival was. Our data suggests that telomerase is an important prognostic indicator of survival in patients with brain tumors. PMID- 16479079 TI - The changes in corneal astigmatism after botulinum toxin-a injection in patients with blepharospasm. AB - To determine if the involuntary contractions of eyelids may have any effects on the development of corneal astigmatism, we performed this prospective study which includes 19 patients with either essential blepharospasm or hemifacial spasm. In hemifacial spasm, the degree of corneal astigmatism was evaluated between two eyes. Then the topographic changes were checked using vector analysis technique before and after passively opening the eyelids. They were also measured before and at 1 and 6 months after the injection of Botulinum toxin. Resultantly, 20 eyes had the with-the-rule (group1) and 9 eyes against-the-rule (group2) astigmatism. In hemifacial spasm, significantly more astigmatism was found at spastic eyes. The corneal topographic changes after passively opening the eyelids showed 10 eyes with the astigmatic shift to the with-the-rule, while the remaining 19 to the against the- rule. At 1 month after injection of Botulinum toxin, group 1 showed reduced average corneal astigmatism, whereas group 2 showed increased astigmatism. The astigmatic change vector showed significantly more against-the-rule. In the contrary, 6 months after treatment, corneal astigmatism again increased in group 1 and decreased in group 2. So they took on the appearance of pretreatment astigmatic status eventually. Conclusively eyelids may play an important role in corneal curvature. PMID- 16479080 TI - Expression of TGFbeta family in the developing internal ear of rat embryos. AB - In order to investigate the expression patterns of the transforming growth factor (TGF)beta isoforms in the internal ear, an immunohistochemical study of rat embryos was performed. Rat embryos were taken on the 13th, 15th, 17th, and 19th day after conception and their internal ears were immunohistochemically stained against TGF beta1, beta2, and beta3. As a result, the 13-day-old embryo showed a very weak positivity to TGF beta1. After the 15th day of pregnancy, no reactivity to TGF beta1 was defected. Immunoreactivity to TGF beta2 was observed from the 15th day of pregnancy throughout the rest of the period. The ampulla of the semicircular canal and the cochlear duct showed a notably strong immunohistochemical reaction. A strong reaction to TGF beta3 was observed on the 15th day of pregnancy. However, no positive reactions were observed thereafter. A strong immunoreactivity was observed especially on the apical cytoplasms, the surfaces of the epithelial cells, and basement membranes of the cochlear duct, as well as the semicircular canals of the developing internal ear of rat embryo. PMID- 16479081 TI - Experience of a Korean disaster medical assistance team in Sri Lanka after the South Asia tsunami. AB - On 26 December 2004, a huge tsunami struck the coasts of South Asian countries and it resulted in 29,729 deaths and 16,665 injuries in Sri Lanka. This study characterizes the epidemiology, clinical data and time course of the medical problems seen by a Korean disaster medical assistance team (DMAT) during its deployment in Sri Lanka, from 2 to 8 January 2005. The team consisting of 20 surgical and medical personnel began to provide care 7 days after tsunami in the southern part of Sri Lanka, the Matara and Hambantota districts. During this period, a total of 2,807 patients visited our field clinics with 3,186 chief complaints. Using the triage and refer system, we performed 3,231 clinical examinations and made 3,259 diagnoses. The majority of victims had medical problems (82.4%) rather than injuries (17.6%), and most conditions (92.1%) were mild enough to be discharged after simple management. There were also substantial needs of surgical managements even in the second week following the tsunami. Our study also suggests that effective triage system, self-sufficient preparedness, and close collaboration with local authorities may be the critical points for the foreign DMAT activity. PMID- 16479082 TI - A fatal case of severe hemolytic disease of newborn associated with anti-Jk(b). AB - The Kidd blood group is clinically significant since the Jk antibodies can cause acute and delayed transfusion reactions as well as hemolytic disease of newborn (HDN). In general, HDN due to anti-Jk(b) incompatibility is rare and it usually displays mild clinical symptoms with a favorable prognosis. Yet, we apparently experienced the second case of HDN due to anti-Jk(b) with severe clinical symptoms and a fatal outcome. A female patient having the AB, Rh(D)-positive blood type was admitted for jaundice on the fourth day after birth. At the time of admission, the patient was lethargic and exhibited high pitched crying. The laboratory data indicated a hemoglobin value of 11.4 mg/dL, a reticulocyte count of 14.9% and a total bilirubin of 46.1 mg/dL, a direct bilirubin of 1.1 mg/dL and a strong positive result (+++) on the direct Coomb's test. As a result of the identification of irregular antibody from the maternal serum, anti-Jk(b) was detected, which was also found in the eluate made from infant's blood. Despite the aggressive treatment with exchange transfusion and intensive phototherapy, the patient died of intractable seizure and acute renal failure on the fourth day of admission. Therefore, pediatricians should be aware of the clinical courses of hemolytic jaundice due to anti-Jk(b), and they should be ready to treat this disease with active therapeutic interventions. PMID- 16479083 TI - Trousseau's syndrome in association with cholangiocarcinoma: positive tests for coagulation factors and anticardiolipin antibody. AB - Thromboembolic events are reported to occur with a high frequency in the setting of malignancy. However, reports on an association between cholangiocarcinoma and pulmonary thromboembolism, thus far, are almost lacking. We present here an unusual case of a 56-yr-old patient presenting cholangiocarcinoma and unexplained pulmonary thromboembolism. The patient had been quite healthy before the diagnosis. Coagulation tests showed elevated levels of fibrinogen, fibrinogen degradation product (FDP), D-dimer, and IgM anticardiolipin antibody (aCL Ab). The thromboemboli were resolved 3 weeks after anticoagulant therapy using low molecular-weight-heparin. Then, follow-up coagulation tests showed a marked decrease to normal in aCL Ab titer as well as the normalization of FDP and D dimer levels. In this case, we describe pulmonary thromboembolism caused by hypercoagulable state associated with cholangiocarcinoma and speculate that such a thrombotic phenomenon could be regressed by anticoagulant therapy. PMID- 16479084 TI - Two cases of isolated diffuse mesangial sclerosis with WT1 mutations. AB - Here we report two cases of isolated diffuse mesangial sclerosis (IDMS) with early onset end-stage renal failure. These female patients did not show abnormalities of the gonads or external genitalia. Direct sequencing of WT1 PCR products from genomic DNA identified WT1 mutations in exons 8 (366 Arg>His) and 9 (396 Asp>Tyr). These mutations have been reported previously in association with Denys-Drash syndrome (DDS) with early onset renal failure. Therefore we suggest that, at least in part, IDMS is a variant of DDS and that investigations for the WT1 mutations should be performed in IDMS patients. In cases with identified WT1 mutations, the same attention to tumor development should be required as in DDS patients, and karyotyping and serial abdominal ultrasonograms to evaluate the gonads and kidney are warranted. PMID- 16479085 TI - Transabdominal embryofetoscopy for the detection of short rib-polydactyly syndrome, type II(Majewski), in the first trimester. AB - Our aim was to demonstrate the potential of first-trimester embryofetoscopy for prenatal diagnosis in a continuing pregnancy. A patient at risk for giving birth to an infant with short rib-polydactyly syndrome, type II (Majewski), presented for prenatal diagnosis at 9 weeks of gestation. A 1 mm semirigid fiberoptic endoscope with an 18 gauge examination sheath and a single-chip digital camera were used for transabdominal embryofetoscopy. Transabdominal embryofetoscopy was performed at 13 weeks of gestation. Direct visualization of the fetus was achieved and no gross limb or facial abnormalities were seen. This case shows that embryofetoscopy is a useful tool for early diagnosis in high-risk patients in the first trimester for continuing pregnancies. PMID- 16479086 TI - Congenital atrophoderma of Pasini and Pierini. AB - Idiopathic atrophoderma of Pasini and Pierini is a form of dermal atrophy of unknown etiology, usually affecting women during their adolescence and young adulthood. A 2-yr-old girl was presented with erythematous atrophic lesion on the right shoulder, which appeared from birth. The histologic findings were consistent with atrophoderma. This patient, to the best of our knowledge, is the first case of atrophoderma with an onset since birth. PMID- 16479087 TI - Giant vascular eccrine spiradenoma: report of a case with immunohistochemical study. AB - We report a rare case of giant vascular eccrine spiradenoma (GVES) which developed in 56-yr-old Korean woman. It is a rare variant of eccrine spiradenoma (ES), which might be mistaken for angiomatous lesions in view of its florid vascularity and hemorrhagic features. Histogenesis of GVES is not clearly elucidated although it is known that ES presumably originates in the eccrine glands. To clarify the histogenesis of GVES, immunohistochemical stainings using various monoclonal antibodies were also performed. The tumor was composed of three types of cells, namely pale epithelial cells, small basal cells, and myoepithelial cells. Therefore, we conclude that GVES originated from eccrine gland and mainly differentiates toward secretory portion of secretory coil. PMID- 16479089 TI - Otolith function assessed with the subjective postural horizontal and standardised stance and gait tasks. AB - If otolith function is essential to maintain upright standing while moving along slanted or uneven surfaces, subjects with an otolith deficit should have difficulty judging whether the inclination of the surface on which they are standing is tilted or not. We tested this judgement and compared it with the ability to control trunk sway during standardised stance and gait tests. Thirteen patients with unilateral vestibular nerve neurectomy at least 6 months prior to testing and 39 age-matched controls were asked to move a dynamic posturography platform on which they were standing back to their subjective 'horizontal' position after the platform had been slowly tilted at 0.4 degrees/s to 5 degrees in 8 different directions. Normal subjects left the platform deviated in pitch (forwards-backwards) at about 0.7 degrees on describing the platform as levelled off for all directions of tilt. Patients showed larger deviations of about 1.3 degrees in pitch with significant differences for forward right tilt (1.58+/-0.73 degrees compared to 0.73+/-0.11 degrees for normals; mean and SEM) and for forward left. Roll (lateral) deviations were about 0.4 degrees for normals and 0.5 degrees larger for the patients (for example, for backward left, 1.13+/-0.24 degrees compared to 0.4+/-0.07 degrees in normals). Except for a tendency towards greater deviations to the lesion side of patients with eyes closed, no differences were noted between tests under eyes open and closed conditions. However, for backward and roll tilts patients needed to steady themselves first by grasping a handrail when tested with eyes closed. Stance tests on foam showed increases in roll and pitch trunk sway with respect to controls. Patients had significantly larger trunk roll sway deviations during 1-legged stance tests and during gait trials. For stance trials, the patients lost their balance control prior to the end of the standard 20-second recording time. We conclude that a unilateral loss of otolith inputs due to nerve resection permanently impairs the ability to judge whether the support surface is horizontal, and leads to excessive trunk sway when standing on a compliant surface as well as excessive trunk roll sway during gait. PMID- 16479088 TI - Hydroxyurea induced perimalleolar ulcers. AB - Hydroxyurea (HU) is an antineoplastic drug commonly used to treat chronic myeloproliferative disorders. Common dermatological side effects include hyperpigmentation, scaling, erythema, alopecia, desquamation of face and hands. Leg ulceration following HU therapy is less common and very few cases have been reported so far. Objective of this paper is to increase the awareness of hydroxyurea induced leg ulcers which will aid in the early diagnosis and appropriate treatment. The first case was a chronic myeloid leukemia (CML) patient on HU 1.5 g/day for 5 yr, who had bilateral painful perimalleolar ulcers for 6 months. The second case was a CML patient on HU 1.5 g/day for 3 yr who developed bilateral lateral malleolar ulcers. Third case was a polycythemia vera (PV) patient on HU 1 g/day for 5 yr who presented with painful medial malleolar ulcer of 2 months. The last case of our report was an elderly PV patient on HU 1.5 g/day for 2 yr and presented with lateral malleolar ulcer which persisted on reducing the dose of HU. In all the 4 cases the ulcers healed on stopping HU. Our report confirms the association of chronic hydroxyurea therapy and perimalleolar ulcers which respond promptly after discontinuation of the drug. The heightened awareness among the physicians will promote early diagnosis and prompt relief from the agonizing ulcers. PMID- 16479090 TI - Intrapartum magnesium sulfate exposure attenuates neutrophil function in preterm neonates. AB - BACKGROUND: Prenatal exposure to magnesium sulfate, a drug that is frequently used for attempted tocolysis in preterm labor, could alter neutrophil functional activity in infants born preterm. OBJECTIVES: To determine the association between maternal tocolysis with magnesium sulfate and the cord blood neutrophil functional activity of preterm neonates. METHODS: The chemotaxis, random motility, and chemiluminescence of neutrophils were compared in the cord blood of 10 preterm neonates born to mothers tocolysed with magnesium sulfate, 10 preterm infants whose mothers had not received any tocolysis, and 10 term infants. Data regarding the maternal and neonatal magnesium and calcium levels were collected and analyzed in association with the cord blood neutrophil functional activity of the preterm infants. RESULTS: Neutrophil functional activity in the cord blood of the preterm neonates was significantly lower than in term neonates. However, the alteration of neutrophil chemotaxis, random motility and chemiluminescence was more noticeable in neonates with intrapartum exposure to magnesium sulfate as compared to preterm infants whose mothers received no tocolysis (30.9 +/- 2.3 vs. 36.7 +/- 2.7 microm, p < 0.01; 26.6 +/- 1.9 vs. 33.1 +/- 3.1 microm, p < 0.01; and 74.3 +/- 6.5 vs. 89.9 +/- 6.25 x 10(3) counts per min (cpm), p < 0.01, respectively). Furthermore, the reduction in neutrophil functional activity of preterm infants with intrapartum exposure to magnesium was directly correlated with the maternal serum magnesium levels (r = -0.90 to -0.85, p < 0.01). CONCLUSION: In infants born preterm, intrapartum exposure to magnesium sulfate is a risk factor contributing to the alteration in neutrophil motility and post phagocytic bactericidal capacity. PMID- 16479091 TI - Dietary insulin affects leucine aminopeptidase, growth hormone, insulin-like growth factor I and insulin receptors in the intestinal mucosa of neonatal pigs. AB - BACKGROUND: Recent studies suggest that milk-borne insulin may regulate the development of the gastrointestinal tract in neonatal mammals. OBJECTIVES: To explore the mechanism by which milk-borne insulin affects gastrointestinal tract development, we examined the effect of dietary insulin on the expression levels of leucine aminopeptidase (LAP) and insulin-like growth factor I (IGF-I), as well as its effect on growth hormone (GH), IGF-I and insulin receptors in the small intestinal mucosa of neonatal pigs. METHODS: Five piglets were anesthetized and sampled within 2-4 h after birth. They were not allowed to suckle and were used as newborn controls (group N). Ten other piglets from 5 litters were randomly divided into group M (n=5), which was fed cow's milk, and group MI (n=5), which was fed cow's milk and insulin (2.5 mg/l). Piglets in groups M and MI were artificially fed for 3 days and then sampled. Total RNA in their intestinal mucosa was extracted with Tripure reagents (Roche, USA). Reverse transcription PCR (RT-PCR) was used to semi-quantify mRNA levels of target genes and 18S rRNA was used in an RT-PCR system as an internal control. PCR products were loaded onto a 9% nondenaturing polyacrylamide gel. The gel was stained by silver staining agents. Digital photos were taken and the strength of the band areas was quantified using software. RESULTS: The results showed that the DNA contents and LAP activity in the small intestines of the piglets in group MI were higher (p<0.05) than in the piglets in group N. Compared with group M, piglets in group MI exhibited significantly increased expression levels of both insulin and GH receptor in the ileum, and LAP in the jejunum (p<0.05); IGF-I receptor expression levels in both the jejunum and ileum were significantly decreased (p<0.01 and p<0.05, respectively), while IGF-I expression was unchanged (p>0.05). CONCLUSION: Collectively, dietary insulin increased mRNA levels of insulin and GH receptor, which could help explain the effect of dietary insulin on receptor-mediated postnatal development of the small intestine. Dietary insulin suppressed IGF-I receptor expression, which may be the result of negative feedback caused when insulin binds to IGF-I receptors. PMID- 16479092 TI - Is the type of hemodialysis important to control serum phosphate? PMID- 16479093 TI - New method for phosphate kinetics estimation during hemodialysis and on-line hemodiafiltration with endogenous reinfusion. AB - AIM: The purpose of this study was to optimize the operative and analytical methodologies to a more exact determination of intradialytic kinetics of the phosphates (P) tested in hemodialysis (HD) and in on-line hemodiafiltration with endogenous reinfusion (HFR - Hemo Filtrate Reinfusion). METHODS: The mass balance measurements of urea and P were carried out in 18 clinically stable HD patients. The effective blood flow (Qb) was measured with a Transonic monitor. The plasma was deproteinized with 10% trichloroacetic acid to prevent breakdown of the proteins and the consequent pseudohyperphosphatemia. Subsequently the supernatant containing the ultrafiltrable phosphates was made to react with a solution of ammonium molybdate for a spectrophotometric reading. RESULTS: The mean urea mass transfer in HD was 16.9 g/session and in HFR 15.4 g/session. The mean P mass transfer in HD was 726 mg/session and in HFR 679 mg/session. Nevertheless, in HFR a significant difference was verified between the clearances of P, between the plasma water side (122.4 +/- 30.8 ml/min) and the dialysate side (105.9 +/- 19.4 ml/min). CONCLUSION: As far as the P mass transfer is concerned, the data obtained is able to be superimposed with that described in the literature during HD, while in HFR it is possible to hypothesize a high efficiency, thanks to an increased output of P in relation to the phenomenon of adsorption which, although is limited, contributes to the transfer of the total mass. Based on this study and re-examining the literature on P kinetics, there is space for methodological improvement both on the operating front with careful determination of the effective Qb, and on the chemical front overcoming the inaccuracy of automatic analyzers in determining the plasma P owing to possible overestimation of phosphatemia and poor sensitivity in measuring the lower levels of P present in the dialysate and/or ultrafiltrate. PMID- 16479094 TI - Hematocrit and prohepcidin: causation or simply association? PMID- 16479095 TI - Plasma prohepcidin positively correlates with hematocrit in chronic hemodialysis patients. AB - BACKGROUND/AIMS: Experimental studies have demonstrated interleukin-6 and iron load induce expression of hepcidin (an iron regulatory peptide), whereas anemia and erythropoietin (EPO) suppress its expression. We are the first to explore the relationships of plasma prohepcidin (the pro-hormone of hepcidin) in patients undergoing chronic hemodialysis (HD). METHODS: We enrolled 71 chronic HD patients. During the preceding 3 months before enrollment, they all had steady weekly levels of haematocrit (Hct) and fixed subcutaneous doses of recombinant human EPO. Plasma levels of prohepcidin, proinflammatory cytokines, and EPO were determined by ELISA kits. RESULTS: Of the patients, prohepcidin levels correlated positively with Hct, and negatively with interleukin-6 and EPO. Examined by a multivariate lineal regression method, we found Hct was the only significant predictor of plasma prohepcidin level. However, prohepcidin had no significant correlation with iron profiles. CONCLUSION: Our findings suggest prohepcidin expression in chronic HD patients might be positively regulated by Hct. PMID- 16479096 TI - Antiplatelet therapy in stroke prevention: present and future. AB - White platelet-fibrin thrombi often form on roughened endothelial surfaces and unstable arterial plaques. Agents that reduce the tendency of platelets to aggregate, agglutinate, and secrete and to attach to endothelial surfaces have been explored as agents that prevent brain and heart infarction. Aspirin, ticlopidine, clopidogrel, dipyridamole, cilostazol, and glycoprotein llb/llla inhibitors are all used now and have various different modes of action and functions. PMID- 16479097 TI - New modalities and aspects of antiplatelet therapy for stroke prevention. AB - Antiplatelet therapy is indicated for secondary prevention of ischaemic stroke. The first-line antiplatelet agent is aspirin. The effect of aspirin is, however, very limited, and this limited effect of aspirin is argued with termed 'aspirin resistance'. Strategies against aspirin resistance may include alternative use of other antiplatelet agents, combination of aspirin with other antiplatelet agents and investigation into molecular targets to develop novel antiplatelet agents. Progress in antiplatelet therapy should be directed at further reducing the risk of ischaemic events including ischaemic stroke without increasing the risk of haemorrhagic events including haemorrhagic stroke. PMID- 16479098 TI - Regional differences in incidence and management of stroke - is there any difference between Western and Japanese guidelines on antiplatelet therapy? AB - PURPOSE: There have not been many discussions on the differences between the guidelines for the management of stroke used in eastern and western countries. The purpose of this paper was to examine whether or not there are substantial differences between western countries and Japan in the prevalence of stroke and the frequencies of stroke subtypes, as well as in the recommended therapy for secondary prevention of ischemic stroke. RESULTS AND CONCLUSIONS: Although there are racial differences and differences in approved drugs between the East and West, the prevalence of stroke and the frequencies of stroke subtypes tend to converge throughout the world. However, the ratio of stroke to ischemic heart disease is still different between the East and West. Comparison of various countries' guidelines shows that recommendations on antiplatelet therapy for secondary prevention of ischemic stroke are fundamentally similar in the East and West, but the recommended doses of antiplatelets, especially aspirin and ticlopidine, are smaller in Japan. Furthermore, Japanese guidelines only recommend the use of antiplatelets (particularly cilostazol) for patients with lacunar infarction with evidence. PMID- 16479099 TI - Antiplatelet therapy after percutaneous coronary intervention. AB - There is intense interest in the relationship between inflammation, thrombosis, platelet aggregation, and hyperlipidemia in patients with coronary artery disease. The specific role of inflammation with its linkage to the coagulation cascade has been well studied. A number of inflammatory markers have been identified which can be used for risk stratification in patients with acute coronary syndromes. Patients with acute coronary syndromes at the time of presentation often have an underlying inflammatory state which needs therapy with antiplatelet regimens including now increasingly frequently clopidogrel in addition to the standard of aspirin. In those patients who are treated medically for their acute coronary syndromes, long-term treatment with dual antiplatelet therapy has been documented to be associated with improved outcome. In patients who undergo an invasive approach with placement of intracoronary stents, the importance of dual antiplatelet therapy is increased. Drug-eluting stents are now used in approximately 90% of all interventional procedures. There is evidence to suggest that while these patients have improved outcome in terms of a decreased need for subsequent procedures to treat restenosis, there is the potential for late subacute stent thrombosis. When late subacute stent thrombosis occurs, it results in mortality or infarction in 40-60% of patients. Dual antiplatelet therapy is therefore recommended for an increasingly longer time in this patient group. At the present time, protocols indicate 3 months for one of the drug eluting stents and 6 months for the other. However, increasingly longer antiplatelet therapy is being used clinically. Assessment of platelet function during follow-up is as yet early. There are issues about which specific test to use and the definition of platelet hyperreactivity. In the future, more individually targeted therapy may be possible if we can more adequately assess the degree of hyperreactivity and underlying inflammation. PMID- 16479100 TI - Polypill strategy vs. prevention clinics for stroke prevention. AB - Thirty years of clinical trials have demonstrated that stroke can be prevented by reducing blood pressure to less than 140/90 mm Hg or by a further decrease of 10/5 mm Hg when blood pressure is controlled, by reducing LDL cholesterol by at least 1 mmol/l (39 mg/dl), and by inhibiting platelet aggregation. The (too simplistic) concept of the polypill strategy combining pharmacologic agents to achieve these goals is supposed to reduce the risk by 80% overall. However, reducing general salt and sugar intake, giving up smoking, developing prevention clinics with prevention nurses to improve adherence to preventive treatments, as well as decreasing poverty and malnutrition, increasing the level of hygiene, fighting against infectious diseases and financially helping developing countries are all combined necessary approaches to decrease the worldwide burden of stroke and other vascular diseases. PMID- 16479101 TI - Dilated cardiomyopathy associated with toluene abuse. AB - The use of paint thinner and glue to achieve an euphoric state has been associated with serious social and health problems in children and young adults. We present the case of a 21-year-old man with dilated cardiomyopathy occurring following abuse of paint thinner and glue containing toluene as main compound. After cessation of toluene abuse, the patient recovered rapidly and completely. Because of the increasing prevalence of toluene abuse, harmful effects of this volatile agent on the heart are also discussed. PMID- 16479102 TI - Deadly myocardial fibrosis of unknown etiology in adolescence. PMID- 16479103 TI - Myocardial infarction in a young African-American male due to myocardial bridging. AB - Myocardial bridging is a clinically uncommon congenital anomaly characterized by tunneling of the coronary artery within the myocardial tissue, usually seen in the left anterior descending artery. Myocardial bridging is associated with altered intracoronary hemodynamics during systole and diastole, determined by the severity and the location of the bridging within the coronary artery. Patients with myocardial bridging may present with angina in the absence of other coronary risk factors which may paradoxically improve with exercise due to an increased intrasystolic pressure, preventing vessel compression. It is uncommon to have bridging in the right coronary artery; it is even more uncommon to have right coronary artery bridging with angina and significant ECG changes. We present a case involving bridging of the right coronary artery with significant symptoms and ECG changes. PMID- 16479104 TI - Effect of tirofiban therapy on ST segment resolution and clinical outcomes in patients with ST segment elevated acute myocardial infarction undergoing primary angioplasty. AB - BACKGROUND: In our study, we assessed the effect of glycoprotein (GP) IIb/IIIa receptor inhibition on microvascular flow after acute coronary occlusion using the early sum of ST segment resolution in electrocardiography. Platelets may play a major role in the dissociation of epicardial artery recanalization and tissue level reperfusion, referred to as the 'no-reflow phenomenon'. Therefore, GP IIb/IIIa receptor inhibition might improve myocardial reperfusion, distinct from its effects on epicardial patency. METHODS AND RESULTS: One hundred and fifteen patients (mean age 57.7 +/- 12.2 years, 96 males, 19 females) with < or = 12-hour acute ST segment elevation myocardial infarction who underwent successful primary percutaneous coronary intervention were retrospectively enrolled into the study. Patients were grouped according to whether they received tirofiban therapy or not. Clinical and electrocardiographic parameters were evaluated. The first sum of ST segment elevation amounts in millimeters was obtained immediately before angioplasty and the second 60 min after restoration of thrombolysis in myocardial infarction III flow. The difference between the two measurements was accepted as resolution of the sum of ST segment elevation and expressed as SigmaSTR. There were no significant differences between the groups regarding age, gender, cardiovascular risk factors, and laboratory parameters, duration from angina onset to the emergency unit, and from door to angioplasty. SigmaSTR was higher in patients who received tirofiban than in those who did not (7.2 +/- 2.8 and 4.2 +/ 2.6 mm, respectively; p < 0.001). There was a significant and positive correlation between GP IIb/IIIa inhibition and SigmaSTR (r = 0.336, p < 0.001), as well as between ejection fraction and SigmaSTR (r = 0.310, p < 0.001). GP IIb/IIIa inhibition was the only independent determinant of SigmaSTR in a multivariate linear regression model which contains 10 variables (p < 0.001). The incidence of in-hospital post-myocardial infarction refractory angina, reinfarction, and heart failure was significantly lower in the tirofiban group (p < 0.05, p < 0.05, and p < 0.05, respectively). Additionally, after 30 days, reinfarction and heart failure were lower in the tirofiban group (p < 0.05 and p < 0.05, respectively). CONCLUSIONS: It is well known that SigmaSTR determines microvascular perfusion. This study shows that GP IIb/IIIa inhibition with tirofiban is of value in preserving microvascular perfusion after restoring coronary thrombolysis in myocardial infarction III flow. PMID- 16479105 TI - A high-resolution single photon emission computed tomography study of verbal recognition memory in Alzheimer's disease. AB - BACKGROUND: In view of the recent technological advances and its ease of availability, we used single photon emission computed tomography (SPECT) to examine the performance of early Alzheimer's disease (AD) subjects on a verbal recognition memory task. METHODS: Eighteen early AD and 10 matched healthy control subjects underwent split-dose (99m)Tc-HMPAO (Ceretec) SPECT using a verbal recognition memory and control task. SPECT images co-registered with MRI scans were used to determine relative regional cerebral blood flow (rCBF) changes in regions of interest. RESULTS: In healthy control subjects, verbal recognition increased rCBF in the right occipital region, thalamus, left prefrontal pole, posterior parietal region and cerebellum, and decreased rCBF in the right hippocampus. AD subjects showed bilateral prefrontal, posterior parietal and occipital increases, unilateral increase in the left posterior temporal region, and bilateral reductions in the hippocampus. Although activation was significantly different between the two groups in the right thalamus and left medial prefrontal region, the verbal recognition task did not enhance discrimination between groups. CONCLUSIONS: Compared with controls, AD subjects activate a similar but more extensive bilateral network during verbal recognition, possibly reflecting an attempt to compensate for impaired processing. PMID- 16479106 TI - Hartmann's operation - still relevant in the 21st century? PMID- 16479107 TI - Chemotherapy for colorectal cancer. AB - Colorectal cancer is the most commonly diagnosed cancer in the EU. Various randomised studies have shown a survival benefit with chemotherapy in the adjuvant setting. Adjuvant chemotherapy with 5-fluorouracil/folinic acid (5FU/FA) for 6 months after curatively resected node-positive colon cancer has become the standard practice. However, controversy still exists regarding the optimal regimen and whether to treat node-negative patients. The latest QUASAR trial results seem to strengthen the argument in favour of adjuvant treatment of Dukes B cancer. Patients with Dukes B tumours and any adverse prognostic indicator should be given the benefit of adjuvant therapy. A number of novel agents (oxaliplatin, irinotecan) showing activity in advanced disease are currently being evaluated in the adjuvant setting. A patient with metastatic colorectal cancer should today be expected to have a median survival of 18-20 months compared to that of 11-14 months only a few years ago. 5FU/FA has been the mainstay of therapy for metastatic colorectal cancer for over 40 years and confers a survival benefit over supportive care. The response rate of 5FU is improved by modulation with FA or by continuous infusional regimens (currently the best expected response rate is around 20-25%). As per the recent National Institute for Clinical Excellence guidelines, the oral agents capecitabine or tegafur with uracil (in combination with FA) can be used as first-line treatment in metastatic colorectal cancer and, although their response rate has not been directly compared to infusional 5FU, survival is unlikely to be inferior. Newer chemotherapeutic agents like irinotecan and oxaliplatin are now entering regular usage due to improved response rates (around 50% in 5FU/FA-containing doublets) and survival. Irinotecan monotherapy is second-line treatment approved by the National Institute for Clinical Excellence, although sequential infusional 5FU/FA irinotecan to infusional 5FU/FA oxaliplatin may convey the best survival with the least side effects. The position of combination chemotherapy before (to downstage) or after metastasectomy (usually from the liver) is still a topic of heated debate. Other routes (intrahepatic, intraperitoneal) are still to be proven and not recommendable outside the trial setting. The latest results of chemotherapy combinations with biological treatments (bevacuzimab and cetuximab) have been very promising indeed. Further improvements in survival, response and quality of life are expected. . PMID- 16479108 TI - Caecal herniation through the foramen of Winslow. PMID- 16479109 TI - Epithelioid hemangioendothelioma of the liver: imaging diagnosis of a rare hepatic tumor. PMID- 16479110 TI - Reversal of Hartmann's procedure after surgery for complications of diverticular disease of the sigmoid colon is safe and possible in most patients. AB - BACKGROUND: Although evidence is growing that most patients who need an operation for diverticular disease of the sigmoid colon can be treated by a single-stage procedure, a two-stage procedure will still be necessary in some patients because of significant sepsis or technical difficulties. The outcomes of 65 patients who underwent secondary restoration after a Hartmann procedure for complicated diverticulitis were studied and the factors leading to complications and mortality were identified. PATIENTS AND METHODS: Of 91 patients, in a consecutive 12-year period, whose primary operation was a Hartmann procedure, 72 survived longer than 3 months after discharge. Sixty-five underwent an attempted reversal of the Hartmann procedure. The POSSUM scores were calculated in all patients as well as the morbidity and mortality rates. RESULTS: In 63 (96.9%) patients the bowel continuity could be restored with a morbidity of 38.5% and a mortality of 3.1%. The POSSUM and p-POSSUM scores adequately predicted the mortality in this series. CONCLUSION: This series shows that when surgical treatment for complicated diverticular disease of the sigmoid colon is necessary, the Hartmann procedure is still a valid indication. In a high percentage of patients the Hartmann procedure could be restored with a low mortality. PMID- 16479111 TI - First results after introduction of the four-armed da Vinci Surgical System in fully robotic laparoscopic cholecystectomy. AB - BACKGROUND: Laparoscopic cholecystectomy offers less post-operative pain, less complications, and faster recovery compared with open cholecystectomy. However, laparoscopic surgery can be demanding because of several technical drawbacks. Robotic surgery allows dexterity skills to be performed faster and shortens the learning curve, possibly leading to faster and safer laparoscopic surgery. METHODS: In this paper, we report the results of our first 12 cases of fully robotic laparoscopic cholecystectomy using the da Vinci Surgical System, comparing them with 12 cases of conventional laparoscopic cholecystectomy. Using a fourth arm in robotic laparoscopy enables the surgeon to perform surgery without the use of a tableside assistant, leading to non-tiring, tremble-free assistance and reducing salary costs. Primary end points are operating time and costs. Secondary end points are operative complications and duration of admission. RESULTS: Fully robotic cholecystectomy was completed in all 12 cases without increased complication rate and without conversions. However, robotic assistance results in an increased overall operating room stay of 31 min and increased costs of EUR 1,180.62. CONCLUSION: Fully robotic laparoscopic cholecystectomy is safe and feasible but seems more expensive and time consuming at this moment. PMID- 16479112 TI - Clinicopathologic characteristics of hepatitis C virus-associated intrahepatic cholangiocarcinoma. AB - BACKGROUND: Although hepatitis C virus (HCV) infection is a possible pathogenetic factor for intrahepatic cholangiocarcinoma (ICC), clinicopathologic features of ICC patients with HCV infection remain unclear. PATIENTS AND METHODS: Of 50 patients with ICC treated, 19 were infected with HCV (HCV-positive group) and 38 underwent surgical treatment. Clinicopathologic features and postoperative outcome were compared between patients with and without HCV infection. RESULTS: In 15 patients in the HCV-positive group, ICC was detected during follow-up for chronic hepatitis C. The proportion of patients who underwent curative resection was significantly higher in the HCV-positive than in the HCV-negative group, and tumors were significantly smaller in the HCV-positive than in the HCV-negative group. In the HCV-positive group, tumors were significantly smaller in patients who were followed up for chronic hepatitis C than in patients who were not followed up. Although cumulative survival rates did not differ significantly between groups, prognoses of patients with small ICC were significantly better than those with large ICC. CONCLUSION: Follow-up for patients with chronic hepatitis C by imaging series at regular intervals is important and provides the possibility to detect a small ICC as well as a hepatocellular carcinoma. Resection should be guided by liver function and the tumor stage in patients with HCV-associated ICC. PMID- 16479113 TI - Laparoscopic staging in gallbladder cancer. AB - PURPOSE: Laparoscopy is beneficial in the staging of pancreatic and upper gastrointestinal malignancies but its role in gallbladder cancer has not been investigated. We evaluated the role of laparoscopy in the staging of gallbladder cancer. METHODS: From 1989 through 2001, 91 patients with gallbladder cancer, without any evidence of metastatic disease on imaging (ultrasound and/or computed tomographic scan), underwent staging laparoscopy. Peritoneal and surface liver metastases were looked for and assessment of local spread was done if possible. Assessment was based on visual impression and biopsies were not obtained routinely. RESULTS: At laparoscopy, 34 (37%) patients had disseminated disease in the form of liver and/or peritoneal deposits; no further surgery was performed in 29 of these patients while 5 patients underwent surgical bypass procedures. Liver metastases were missed at laparoscopy in 2 patients and were subsequently found at laparotomy. Assessment of the gallbladder mass was possible in 33 (36%) patients, 6 of these were found to have extensive local disease and did not undergo any further surgery. Laparoscopic staging, thus avoided further surgery in 35 (38%) patients. Of the 51 patients without metastatic disease, who underwent laparotomy, 11 were found to have nonresectable locally advanced disease while 1 had liver metastases, which were missed at laparoscopy; 7 underwent bypass procedures only; 21 underwent simple cholecystectomy and extended cholecystectomy was done in 11 patients. The resectability rate (number of resections/operations) in patients undergoing laparoscopic staging was 57% (32/56) as compared with 43% (142/328) in those who did not undergo laparoscopy. CONCLUSIONS: Staging laparoscopy in patients with gallbladder cancer detected liver and peritoneal metastases that were missed on imaging. It reduced the number of unnecessary surgical explorations and improved the resectability rate. PMID- 16479114 TI - External pancreatic fistula as a sequel to management of acute severe necrotizing pancreatitis. AB - BACKGROUND/AIMS: External pancreatic fistula (EPF) is a common sequel to surgical or percutaneous intervention for infective complications of acute severe pancreatitis. The present study was aimed at studying the clinical profile, course and outcome of patients with EPF following surgical or percutaneous management of these infective complications. METHODS: A retrospective analysis of clinical data of patients with EPF following intervention (surgical or percutaneous) for acute severe pancreatitis managed between January 1989 and April 2002 recorded on a prospective database was done. Univariate analysis of various factors (etiology, imaging findings prior to intervention, fistula characteristics and management) that could predict early closure of fistula was performed. RESULTS: Of 210 patients with acute severe pancreatitis, 43 (20%) patients developed EPF (mean age 38 (range 16-78) years, M:F ratio 5:1) following intervention for infected pancreatic necrosis (n=23) and pancreatic abscess (n=20) and constituted the study group. The fistula output was categorized as low (<200 ml), moderate (200-500 ml) and high (>500 ml) in 29 (67%), 11 (26%) and 3 (7%) patients, respectively. Fifteen patients (35%) had morbidity in the form of abscess (n=5), bleeding (n=1), pseudoaneurysm (n=2) and fever with no other focus of infection (n=7). Spontaneous closure of the fistula occurred in 38 (88%) patients. The average time to closure of fistula was 109+/- 26 (median 70) days. Fistula closed after intervention in 5 patients (2 after endoscopic papillotomy, 1 after fistulojejunostomy and 2 after downsizing the drains). Of the 38 patients with spontaneous closure, 9 (24%) patients developed a pseudocyst after a mean interval of 123 days of which 7 underwent surgical drainage of the cyst. Univariate analysis of various factors (etiology, imaging findings prior to intervention, fistula characteristics and management) failed to identify any factors that could predict early closure of fistula. CONCLUSIONS: EPF is a common sequel following intervention in acute severe pancreatitis. The majority of these are low output fistulae and close spontaneously with conservative management. One fourth of patients with spontaneous closure develop a pseudocyst as a sequel, requiring surgical management. PMID- 16479115 TI - The "Protocole d'Examen Cognitif de la Personne Agee - Lausanne" (PECPA-L): a cognitive assessment tool for the French-speaking elderly in Switzerland. AB - OBJECTIVE: The main objective of this study was to validate and normalize, in the French-speaking population of Switzerland, an empirically derived structured cognitive assessment scale for the elderly. METHOD: 237 healthy elderly and 115 elderly with mild-to-moderate dementia as well as a preliminary sample of 27 subjects with MCI, all community-dwelling, were assessed using the PECPA-L for its validation and normalization using Crohnbach's alpha, rank comparisons, and AUROC measures. RESULTS: The internal consistency of the PECPA-L was good (Crohnbach's Alpha=0.79). The discriminating power of the PECPA-L between the normal elderly and the mildly to moderately demented (AUROC=0.940) and those with MCI (AUROC=0.925) was high. Normal aging does not influence all cognitive domains equally; therefore, the subscores of the PECPA-L vary in their discriminating power between the normal and the demented elderly, with gnosis abilities having the least (AUROC=0.719) and delayed memory performance having the highest (AUROC=0.927) discriminating power. Normative data according to age and education are provided. CONCLUSIONS: The PECPA-L is a highly appropriate tool for the detection and documentation of early cognitive impairment in the French-speaking population in Switzerland while accounting for age and education. PMID- 16479116 TI - Hyperechoic plaque: an ultrasound marker for osteoporosis in acute stroke patients with carotid disease. AB - BACKGROUND: Osteoporosis is a significant complication of stroke, and hip fracture after a stroke is a frequent problem. Moreover, growing evidence links vascular and bone diseases, in the form of osteoporosis associated with both atherosclerosis and vascular calcification. The aim of our study is to detect bone change in the acute phase of ischemic stroke in patients with carotid disease and to verify the correlation with carotid echogenic plaques. PATIENTS AND METHOD: Out of 245 subjects consecutively admitted to our Stroke Unit for their first ischemic stroke, we selected 49 patients with a first-ever stroke due to carotid atherosclerosis without a previous diagnosis of bone disease. We assessed risk factors for cerebrovascular disease as well as for osteoporosis, the degree of neurological deficit and disability, and bone mineral density that was quantified by bilateral hip dual energy X-ray absorbimetry. Osteoporosis was defined as a T score below -2.5. Carotid ultrasound was used to classify plaques in non-hyperechoic (grade 1) and hyperechoic plaque (grade 2). RESULTS: We found a high prevalence of low bone mass density (BMD) in our patients (18 out of 49=36.7%), without relationship to the side of paresis. According to univariate analysis evidence of osteoporosis was correlated with age (p=0.05), score of Scandinavian Stroke Scale (p=0.01) and grade 2 plaque (p=0.01). According to multivariate analysis, there was a significant positive correlation between grade 2 plaques and osteoporosis (OR=6.58; 95% CI=1.57-27.54; p=0.01), which was stronger in women (OR=18.15; 95% CI=1.80-182.83; p=0.01). The percentage of intraplaque hyperechogenicity was inversely correlated with BMD (r=-0.411, p=0.016). CONCLUSION: Osteoporosis is highly prevalent in acute atherosclerotic stroke patients. Carotid hyperechoic plaque is an independent marker of osteoporosis. PMID- 16479117 TI - A case of Melas (A3243G) on chronic dichloroacetate treatment. PMID- 16479118 TI - Hydrocephalus and hyponatremia as the presenting manifestations of primary CNS lymphoma. PMID- 16479119 TI - Hepatolenticular degeneration combined with primary antiphospholipid syndrome: a case report. PMID- 16479120 TI - Progressive multifocal leukoencephalopathy in a patient with hepatosplenic T cell lymphoma. PMID- 16479122 TI - Glossopharyngeal neuralgia. AB - Glossopharyngeal neuralgia is a distinctive syndrome, named by Wilfred Harris. Investigation must exclude multiple sclerosis, and local compression, especially by tumours which require treatment. Dandy deserves credit for first indicating vascular compression of cranial nerve roots as a cause of cranial neuralgias, and Jannetta for establishing neurovascular decompression. Vascular compression is a common and treatable cause but does not account for all previously designated idiopathic cases. PMID- 16479121 TI - Hemimedullary syndrome with ipsilateral sensorimotor deficits. PMID- 16479123 TI - Sir William Withey Gull (1816-1890). AB - William Gull, a man of humble origins, became one of the outstanding, polymath physicians of the 19th century, and physician to Queen Victoria. Among several important and original contributions, this paper summarises his work on myxoedema, paraplegia and anorexia nervosa. PMID- 16479124 TI - A Norse contribution to the history of neurological diseases. AB - Multiple sclerosis (MS) is prevalent in areas with many inhabitants of Scandinavian descent, and a 'Viking gene' hypothesis has been suggested for the dissemination of the disease. It is therefore relevant to search Norse sagas for descriptions of clinical pictures which could have been MS. The saga of Bishop Thorlak describes a woman named Halldora, who suffered from transient paresis between 1193 and 1198. The diagnosis is uncertain, but the story shows that symptoms associated with MS were known in Iceland at the end of the 11th century. PMID- 16479125 TI - Neuralgia, neurology, neuro-. PMID- 16479127 TI - Does pharmaconutrition with L-arginine and/or alpha-tocopherol improve the gut barrier in bile duct ligated rats? AB - BACKGROUND/AIM: Nitric oxide supplementation and antioxidant therapy modulate gut barrier function, but the relationships between enhanced nitric oxide production, antioxidant administration, and biliary obstruction remain unclear. We evaluated the role of nitric oxide and alpha-tocopherol supplementation in bile duct ligated rats. METHODS: Fifty male Wistar albino rats underwent sham operation (group I; control animals) or bile duct ligation (groups II, III, IV, and V). The ligation groups received the following regimens: standard pellet diet (group II), pellet diet plus intramuscularly administered alpha-tocopherol (group III), and L arginine-enriched pellet diet without (group IV) or with (group V) alpha tocopherol. Nitric oxide, malondialdehyde, and alpha-tocopherol concentrations were assessed at the end of 3 weeks. Liver and intestinal samples were scored histologically. Mesenteric lymph node and liver cultures were assessed for bacterial translocation. RESULTS: The liver malondialdehyde concentration was highest in group III. The nitric oxide content in the liver was higher in groups III and V, as were the blood alpha-tocopherol levels. Bacterial translocation was evident following bile duct ligation, but did not differ among the treatment groups. Intestinal histology revealed that group III had the lowest villus height, that group V had the least villus count, and that group II had the highest mucous cell count. The fibrosis scores were higher in groups IV and V. CONCLUSIONS: An obvious effect of alpha-tocopherol (with or without L-arginine) on the gut barrier could not be demonstrated. Moreover, the L-arginine-enriched diet promoted fibrosis in the liver. Thus, while biliary duct obstruction triggers bacterial translocation, nitric oxide and/or alpha-tocopherol supplementation did not seem to improve the gut barrier in our model. PMID- 16479128 TI - Reversible cholestasis and cholangitis induced by biliary drainage and infusion in the rat. AB - To develop a rat model of ascending cholangitis, we constructed a controllable and accessible biliary drainage and infusion system. We first modified a reversible cholestasis model of the rat and then induced ascending cholangitis by administration of Escherichia coli into the proximal choledochostomy tube. After biliary infusion of E. coli, the liver, choledochostomy tube and bile were all positive for E. coli, but no bacteria grew in rats receiving biliary infusion of normal saline. Retrograde cholangiography of the initial choledochostomy ensured that the tube end was in the right position in the proximal common bile duct. The patency of the tube-tube choledochocholedochostomy was confirmed by a cholangiogram on day 90. Thirty days after the tube-tube choledochocholedochostomy, the livers of the experimental animals did not differ from the control livers. The tube-tube choledochocholedochostomy model not only provides reproducible, reliable, reversible cholestasis, but creates a sustainable and accessible biliary infusion system. This can be used for long term investigations of repeated cholangitis and recurrent cholestasis. PMID- 16479130 TI - Prevalence of voice complaints, risk factors and impact of voice problems in female student teachers. AB - A cross-sectional questionnaire survey was done among 457 female student teachers and 144 females in the general population. The conclusions are based on the opinions of student teachers and the general population. The results of this study show that 39.6% of the student teachers and 32.6% of the general population reported voice complaints at the moment and/or over the past year (p=0.198). The association between various risk factors (vocal loading factors, physical factors, environmental factors and psycho-emotional factors) and voice complaints were examined. Vocal load was reported in both the student teachers and the general population (p=0.322). Among the subjects with voice complaints, the student teachers were significantly more of the opinion than the general population that environmental irritants in the classroom (p=0.001) and the composition of the group they communicate with (p=0.033) have a negative influence on their voice. In the groups with voice complaints, the student teachers reported significantly less than the general population that stress (p=0.004) and the deterioration of their general physical condition (p=0.003) have a negative influence on their voice. Remarkably, over a third of the student teachers and one fifth of the general population with voice complaints were of the opinion that decrease of hearing has a negative influence on their voices (p=0.113). There was no significant difference in Voice Handicap Index (VHI) scores (p=0.284) and impact of voice complaints among student teachers and the general population. Over 15% of the student teachers and the general population with voice complaints reported being or having been disabled due to the voice problem, probably reflecting the severity of the voice problem (p=0.838). The groups reporting voice complaints and disability in relation to their voice complaints have significantly higher VHI scores than those without voice complaints and disability, which indicates a higher psychosocial impact of voice complaints. Only around a third of the student teachers and the general population with voice complaints sought paramedical care (p=0.656)/treatment (p=0.361) for their voice complaint. Only a minority of student teachers (18.6%) and the general population (29.5%) with voice complaints were of the opinion that the number of people they communicate with has a negative influence on their voice (p=0.120). Only around a third of the student teachers and less than a tenth of the general population with voice complaints were of the view that they would develop a voice complaint due to their profession (p=0.003). Less than half of the student teachers and less than one fifth of the general population with voice complaints were aware of the potential risks of their profession on their voice (p=0.002). Voice complaints appear to have a multifactorial genesis. The student teachers are not sufficiently aware of the impact of the various risk factors on their voice. Furthermore, they are not aware of the potential risk that future teaching may have on their voice. This apparent lack of awareness in student teachers may be considered a risk factor for voice complaints. PMID- 16479129 TI - Monitoring of lung edema by microwave reflectometry during lung ischemia reperfusion injury in vivo. AB - It is still unclear whether lung edema can be monitored by microwave reflectometry and whether the measured changes in lung dry matter content (DMC) are accompanied by changes in PaO2 and in pro- to anti-inflammatory cytokine expression (IFN-gamma and IL-10). Right rat lung hili were cross-clamped at 37 degrees C for 0, 60, 90 or 120 min ischemia followed by 120 min reperfusion. After 90 min (DMC: 15.9 +/- 1.4%; PaO2: 76.7 +/- 18 mm Hg) and 120 min ischemia (DMC: 12.8 +/- 0.6%; PaO2: 43 +/- 7 mm Hg), a significant decrease in DMC and PaO2 throughout reperfusion compared to 0 min ischemia (DMC: 19.5 +/- 1.11%; PaO2: 247 +/- 33 mm Hg; p < 0.05) was observed. DMC and PaO2 decreased after 60 min ischemia but recovered during reperfusion (DMC: 18.5 +/- 2.4%; PaO2: 173 +/- 30 mm Hg). DMC values reflected changes on the physiological and molecular level. In conclusion, lung edema monitoring by microwave reflectometry might become a tool for the thoracic surgeon. PMID- 16479131 TI - The prevalence and risk factors for occupational voice disorders in teachers. AB - OBJECTIVE: Occupational voice disorders in Poland account for over 25% of all occupational diseases. The aim of the study was to assess the prevalence of voice problems in the general population of Polish teachers, and identify risk factors for developing voice pathology. PATIENTS AND METHODS: The study group comprised 425 female full-time teachers (most of them primary and secondary school, age ranging from 23 to 61 years) and 83 non-teacher women (control) whose jobs did not involve vocal effort, matched for age to the study group. All participants were subjected to a survey using an extensive questionnaire, and to laryngological, phoniatric and videostroboscopic examinations. RESULTS: The overall lifetime vocal symptoms were more frequent in the teachers than in the non-teachers (69 vs. 36%), and in particular it related to permanent and recurrent hoarseness, and dryness in the throat. Mean number of the voice symptoms was 3.21 in teachers and 1.98 in controls (p < 0.001). Abnormal (non euphonic) voice, neck muscle hypertension during phonation and incorrect resonator function were also significantly more frequent in the teachers. Mean maximum phonation time was shorter in teachers than in the controls (14.3 vs. 15.9 s, p < 0.01). Occupational voice disorders and hyperfunctional dysphonia (that is thought to predispose to such pathology) were found in 32.7% of teachers and 9.6% of control subjects. The probability of developing incomplete glottal closure (odds ratio 13.2x; 95% CI: 1.8-96.8) and hyperfunctional dysphonia (odds ratio 2.7; 95% CI: 1.14-6.44) were significantly higher in the teacher group versus non-teachers. A significant positive relationship was found in teachers between the prevalence of hyperfunctional dysphonia and strained phonation, neck muscle hypertension, instability of voice, self-assessed hyper-arousal, and lifetime vocal effort index (years of employment as a teacher x hours of professional activity/week). The prevalence of vocal nodules and incomplete glottal closure were correlated with incorrect phonation technique parameters, but not with psychological factors. No correlation was found with environmental variables, such as classroom temperature, humidity, airborne dust. CONCLUSION: The prevalence of self-reported symptoms and clinical signs of voice disorders is around 2-3 times more frequent in Polish female teachers than in non-teachers. Lifetime vocal effort, incorrect technique of phonation and psychological predisposition seem to constitute major risk factors for developing occupational voice disorders. PMID- 16479132 TI - Estimation of the voice source from speech pressure signals: evaluation of an inverse filtering technique using physical modelling of voice production. AB - OBJECTIVE: The goal of the study is to use physical modelling of voice production to assess the performance of an inverse filtering method in estimating the glottal flow from acoustic speech pressure signals. METHODS: An automatic inverse filtering method is presented, and speech pressure signals are generated using physical modelling of voice production so as to obtain test vowels with a known shape of the glottal excitation waveform. The speech sounds produced consist of 4 different vowels, each with 10 different values of the fundamental frequency. Both the original glottal flows given by physical modelling and their estimates computed by inverse filtering were parametrised with two robust voice source parameters: the normalized amplitude quotient and the difference (in decibels) between the levels of the first and second harmonics. RESULTS: The results show that for both extracted parameters the error introduced by inverse filtering was, in general, small. The effect of the distortion caused by inverse filtering on the parameter values was clearly smaller than the change in the corresponding parameters when the phonation type was altered. The distortion was largest for high-pitched vowels with the lowest value of the first formant. CONCLUSIONS: The study shows that the proposed inverse filtering technique combined with the extracted parameters constitutes a voice source analysis tool that is able to measure the voice source dynamics automatically with satisfactory accuracy. PMID- 16479133 TI - Comparison of speaking rate, articulation rate and alternating motion rate in dysarthric speakers. AB - Despite recent indications that speaking rate, articulation rate and alternating motion rate (AMR) are sensitive to the influence of neurological disease on speech production in dysarthric speakers, the relationship among these parameters remains largely questionable. In the current study, speech samples were collected from 62 dysarthric speakers and 34 controls while reading a passage. Acoustic analysis investigated speaking rate, articulation rate and AMR and compared these parameters between the dysarthric and control groups and within the control and dysarthric group. The major findings were: (1) Speaking rate, articulation rate and AMR were all markedly lower in dysarthric speakers than in controls. (2) Marked correlations were recognized between speaking rate, articulation rate and AMR in the dysarthric group. (3) Compared with the speaking rate and articulation rate, AMR is characterized as follows: (i) variation was more limited in controls, and (ii) the control group was clearly distinguishable from the dysarthric group. (4) Unlike in controls, AMR in the dysarthric speakers was notably lower than their articulation rate. We therefore conclude that AMR is a more easily detected sign of abnormal articulation than speaking rate and articulation rate. PMID- 16479134 TI - Reproducibility of the Dutch version of the Voice Handicap Index. AB - OBJECTIVE: To investigate the reproducibility of the Dutch translation of the Voice Handicap Index (VHI) by performing a test-retest study. To determine the relationship between the test-retest differences and the VHI scores. PATIENTS AND METHODS: A group of 104 patients (56 male, 48 female) with voice complaints completed the VHI twice with a mean interval of 13 days. RESULTS: There were no differences in scores between male and female patients. There was a good correlation between the first and the second measurement (r=0.95). The difference between two VHI scores of any one patient and the total score were not related. We found that a 14-point difference in total score of the Dutch VHI for 1 patient at two points in time is significant. CONCLUSION: The Dutch version of the VHI has a good reproducibility. When used for clinical evaluation studies of patients with voice complaints, a difference of 14 points between two measurements is significant. PMID- 16479135 TI - Postthyroplasty implant extrusion. AB - The case of a 57-year-old gentleman with a left-sided vocal cord palsy following a surgical procedure done elsewhere is being reported. His main problem was dysphonia, and speech therapy did not improve his voice much. A thyroplasty type 1 was done to improve his voice. A Silastic implant was inserted for medialization of vocal cords with a good postoperative result. The patient was followed up in the clinic and presented again 7 months after surgery with extrusion of the Silastic implant. Quite remarkably, his voice did not deteriorate even after the extrusion of the Silastic implant. This could be due to formation of fibrous tissue holding the cords in the medial position. The patient retained an excellent voice, and no further procedure was necessary. PMID- 16479136 TI - Postmenopausal bleeding: a diagnostic approach for both private and public sectors. AB - OBJECTIVE: To assess the optimal diagnostic approach to women with postmenopausal bleeding by comparing transvaginal ultrasound and endometrial sampling with office hysteroscopy. METHODS: A prospective collective study was performed on 102 consecutive patients with postmenopausal bleeding who were evaluated by ultrasound measurement of the endometrial thickness (EL), endometrial sampling by Accurette and outpatient hysteroscopy and directed biopsy. RESULTS: Accurette was inadequate for histological diagnosis in 65 of 136 samples and many repeat investigations were required. In all 16 cases of endometrial polyps, ultrasound measurement of the EL was > or =4 mm and hysteroscopy confirmed the findings. Accurette detected only 5 polyps. All 7 cases of endometrial hyperplasia were detected using an EL of > or =4 mm and hysteroscopy confirmed the findings. Accurette failed to detect 5 out of 7 cases of endometrial hyperplasia. Five cases of endometrial cancer were diagnosed: all had an EL > or =4 mm but were inadequately sampled for diagnostic purposes in 3 cases. A definitive diagnosis was made on hysteroscopy in 4 cases (1 patient did not have a hysteroscopy). CONCLUSION: Accurette is not a good sampling device for the diagnosis of postmenopausal bleeding. An ultrasound measurement of the EL > or =4 mm in patients with postmenopausal bleeding warrants further investigation. Outpatient office hysteroscopy is an accurate and sensitive modality to employ as a first line investigation for definitive diagnosis. PMID- 16479137 TI - Endometrial adenocarcinoma presenting with an osseous metastasis. AB - BACKGROUND: Gynecologic cancers metastatic to bone are rare. Endometrial carcinoma usually presents with vaginal bleeding. CASE REPORT: A 67-year-old woman presented with pain, erythema and swelling of the right foot and no history of postmenopausal bleeding. Biopsy revealed primary endometrioid carcinoma metastatic to the calcaneus, talus and metatarsal bones. Lower leg amputation, total abdominal hysterectomy, bilateral salpingo-oophorectomy and pelvic lymph node sampling were performed. Postoperatively the patient received cisplatin with adriamycin and megestrol acetate and is alive with no evidence of disease 20 months after the diagnosis. CONCLUSION: Endometrial carcinoma can present as a metastatic lesion of bone. PMID- 16479138 TI - Effects of intranasal estradiol treatment on serum paraoxonase and lipids in healthy, postmenopausal women. AB - BACKGROUND/AIMS: Serum lipid concentrations worsen after the menopause because of estrogen deficiency, leading to an increased atherogenic pattern. It is known that serum paraoxonase (PON1) activity prevents the development of atherosclerosis. The aim of this cross-sectional study was to observe the effects of intranasal 17beta-estradiol (300 microg/day) on serum PON1 and lipid levels in healthy postmenopausal women. METHODS: 48 healthy, postmenopausal women were enrolled into this cross-sectional study. 28 subjects without an intact uterus and ovaries were using single-dose (300 microg/day) intranasal 17beta-estradiol and 20 subjects with spontaneous natural menopause were not on any hormone therapy. Body mass index (BMI), blood pressure, serum follicle-stimulating hormone, estradiol, fasting glucose, insulin, lipid fractions and PON1 levels were measured. Homeostasis model assessment (HOMA-R) was used to estimate insulin resistance. RESULTS: The higher estradiol, high-density lipoprotein and salt stimulated paraoxonase (SSP) levels were observed in intranasal 17beta-estradiol users in comparison with non-users. There were no statistically significant differences in BMI, blood pressures, other lipid fractions, basal paraoxonase, arylesterase, fasting glucose and insulin levels, HOMA-R between the groups. SSP was inversely associated with fasting insulin levels and HOMA-R. CONCLUSION: These observations may suggest that intranasal 17beta-estradiol does not have harmful effects on the PON1 activity and lipid metabolism. PMID- 16479139 TI - Insulin-lowering agents inhibit synthesis of testosterone in ovaries of DHEA induced PCOS rats. AB - BACKGROUND: Insulin-lowering agents are reported to be useful in treating polycystic ovary syndrome (PCOS) anovulation. It has been suggested that lower insulin levels secondarily affect ovarian tissue, although the direct mechanism of action has not yet been verified. Here we investigated if these agents directly affect the ovary. METHODS: Thirty female Wister rats were studied. Six control rats were injected subcutaneously with 0.2 ml sesame oil, while 24 rats used as PCOS models were injected subcutaneously with dehydroepiandrosterone (DHEA) and divided into four groups. Six rats were injected with only DHEA, while the remaining 18 rats received metformin, pioglitazone or troglitazone. The ovaries were immunohistochemically stained with anti- testosterone and anti 17beta-HSD antibodies, and then evaluated for morphological changes. RESULTS: In the DHEA administration group, the number of atretic follicles significantly increased compared to that of control rats. The insulin-lowering agents did not improve the multicystic appearance. Serum testosterone concentrations significantly increased with DHEA administration, but the increase was inhibited by oral administration of insulin-lowering agents. Testosterone deposits in ovarian tissue were also reduced by feeding rats insulin-lowering agents. CONCLUSION: Insulin-lowering agents affected ovarian tissue by inhibiting testosterone biosynthesis in vivo. PMID- 16479140 TI - Vaginal ultrasonographic and hysterosonographic evaluation of the low transverse incision after caesarean section: correlation with gynaecological symptoms. AB - AIM: We investigated whether there is a correlation between morphological changes of the lower uterine segment after caesarean section (CS), visualized by means of either a transvaginal sonography (TVS) or a sonohysterography (SHG), and the frequency of abnormal uterine bleedings reported by the women. METHODS: By means of a random selection of our population, anamnesis, medical records, and TVS and SHG images of the lower uterine segment were collected in 217 women (116 with previous CS and 101 with previous vaginal birth), and an observational case control study was performed. RESULTS: The uterine incision was identified in almost all women after CS (102/116) using TVS. It was observed that abnormal uterine bleeding was significantly more frequent in the CS group in comparison with the group of women who delivered vaginally. A correlation between the presence of abnormal uterine bleeding and the presence of significant sonographic findings in the lower uterine transverse incision in the women after CS was found. In the CS group, TVS findings were confirmed by those obtained by SHG, and, with this technique, a triangular anechoic area at the presumed site of incision (the niche) was identified in 69 of the 116 women (59.5%). CONCLUSIONS: In this study, we found a correlation between abnormal uterine bleeding and sonographic findings in women after CS. This correlation appears to be more significant in women who had CS 5-10 years ago. A significant difference exists between the CS group and the group of women who delivered vaginally for both frequency of abnormal uterine bleeding and sonographic findings. PMID- 16479141 TI - Role of latent genital tuberculosis in repeated IVF failure in the Indian clinical setting. AB - Genital tuberculosis is reported to be a major pelvic factor causing infertility in Indian women and often exists without any apparent signs and symptoms. The role of latent tuberculosis in repeated IVF failure in unexplained infertility is examined. 81 women with unexplained infertility having repeated IVF failure tested for Mycobacterium tuberculosis using PCR, ZN staining and BACTEC-460 culture were selected. Fresh IVF-ET or frozen embryo transfer (FET) was attempted on patients successfully treated with anti-tubercular drugs (ATD). ATD-treated fresh cycles (group A1) and frozen cycles (group B1) were compared to previously failed fresh cycles (group A2) and FET attempts (group B2), respectively. Main outcome measures were gonadotropin required, terminal E2, number of oocytes retrieved, fertilization rate, embryo quality, endometrial thickness and sub endometrial blood flow (V(max)). Gonadotropin required in group A1 was significantly less as compared to group A2. Number of oocytes retrieved and grade I embryos, endometrial thickness and V(max) were significantly higher in group A1. Endometrial thickness and V(max) were significantly increased in group B1 as compared to B2. The study indicates that latent tuberculosis should be considered in young Indian patients presenting with unexplained infertility with apparently normal pelvic and non-endometrial tubal factors and repeated IVF failure. PMID- 16479143 TI - Phagocytosis of human retinal pigment epithelial cells: evidence of a diurnal rhythm, involvement of the cytoskeleton and interference of antiviral drugs. AB - Retinal pigment epithelial (RPE) cells provide crucial functions for the maintenance of the retinal environment. We investigated the phagocytotic mechanisms of RPE cells evaluating the question whether particle uptake underlies a diurnal rhythm. Additionally, a possible connection of volume regulation and the phagocytotic function of RPE cells was studied. As antiviral nucleoside analogues influence cell-volume-regulating mechanisms, we tested several antiviral drugs. Cultured primary RPE cells and a permanent cell line (ARPE-19) were tested for uptake of europium-labeled microspheres quantified by time resolved fluorometry. Cells were also exposed to cyclic illumination or continuous light and dark culture conditions. Inhibitors of cytoskeleton (microtubuli, actin) and osmotic swelling were also tested. Ingested FITC-labeled microparticles were found in phagosomes strongly associated which the cytoskeleton as they could not be easily moved by laser tweezer microscopy. Phagocytosis was observed predominately during dark intervals and was reduced by continuous light exposure. The diurnal rhythm of unsynchronized RPE cultures was abolished by microtubule inhibitors although no inhibition of overall particle uptake by cytoskeletal blockers was observed. Hypoosmotic swelling of RPE also decreased phagocytosis. Acyclovir was found inhibitory in ARPE-19 cells, whereas azidothymidine showed a protracted inhibiting activity on primary RPE cells and ganciclovir was inactive in both cell types. The presence of a diurnal rhythm also in culture indicates genetic determination of light-regulated particle uptake. This mechanism appears to be influenced by the regulation of cell volume and microtubule function. Inhibition of RPE function by antiviral drugs is a novel finding and in accordance with interferences of the tested drugs with cellular chloride channels described earlier. It may give a hint towards possible ocular side effects in the long-term use of nucleoside-analogous substances. PMID- 16479142 TI - Complete clinical remission and disappearance of liver metastases after treatment with somatostatin analogue in a 40-year-old woman with a malignant insulinoma positive for somatostatin receptors type 2. AB - Insulinoma is the most common pancreatic endocrine tumor, accounting for 40% of all pancreatic functional neoplasm, and is characterized by hypersecretion of insulin and hypoglycemia. Elective treatment for insulinomas is surgical enucleation. Medical therapy with diazoxide, followed by somatostatin analogues in some cases, may be necessary to treat the hypoglycemic symptoms. We report a case of a patient affected by metastatic insulinoma with severe hypoglycemia. After surgery, histopathology confirmed the presence of a malignant insulinoma with multiple metastases in the liver. Due to the persistence of hypoglycemia, the patient was started on octreotide LAR treatment, which determined a complete clinical remission with regression of the metastatic lesions in the liver after one year. Repeated CT scans 2 and 3 years after surgery confirmed the remission. To our knowledge, the complete regression of the disease in insulinomas treated with long-standing somatostatin analogue therapy has never been reported. Immunohistochemical analysis in tissue specimens showed a strong membrane immunoreactivity for somatostatin receptors type 2 (SSTR2) in both the primary nodule and the metastases. The capacity of somatostatin analogues to negatively regulate cell proliferation through indirect and direct mechanisms has been experimentally demonstrated. Furthermore, SSTR2 activation may exert pro apoptotic effects in neoplastic cells. Thus, both mechanisms may have been responsible of the remission of the disease in this patient. This case underlies the potential impact of the treatment of pancreatic insulinomas with somatostatin analogues, and, if confirmed, the usefulness of SSTR determination in these neoplastic specimens. PMID- 16479144 TI - Topical application of mitomycin C in combination with dexamethasone: effective delay of myringotomy closure. AB - The object of this study was to investigate the efficacy of topical mitomycin C (MMC) in combination with subsequent topical dexamethasone to prolong the patency of myringotomies. Bilateral instrumental myringotomies were performed in 12 rats. On both sides, a solution of MMC (2 mg/ml) was applied to the intact tympanic membrane for ten minutes before myringotomy. Dexamethasone (4 mg/ml) was applied to one tympanic membrane of each animal at days 3, 6, 9 and 14. Tympanic membranes were observed weekly for a total of 9 weeks. The mean healing period was found to be 17.5 days (range 4.5-45.5 days) in the group with MMC alone, and 32 days (range 17.5-59.5 days) in the group MMC + dexamethasone. The difference was significant. A combination of topical MMC and subsequent dexamethasone did delay the healing rate of myringotomies in rat tympanic membranes longer than MMC alone. PMID- 16479145 TI - Cervical lymph node metastases from occult squamous cell carcinoma: analysis of 82 cases. AB - PURPOSE: The aim of this study was to analyze the prognostic value of some clinical factors and to compare the survival of different treatment plans in patients with cervical lymph node metastases from occult squamous cell carcinoma (SCC). METHODS: A retrospective review was conducted of patients who were diagnosed as having cervical lymph node metastases from occult SCC. Overall cumulative survival was analyzed using the standard Kaplan-Meier method. Tests of significance were based on log-rank statistics. RESULTS: The 82 patients in the study consisted of 69 males (84.2%) and 13 females (15.8%). The average age at diagnosis was 64.7 years. Fifty patients (60.9%) underwent surgical treatment of cervical metastasis. Radiotherapy was performed in 79 patients. Thirty-two patients (40.5%) received primary fractioned external beam radiotherapy; 47 patients (59.5%) received postoperative fractioned external beam radiotherapy. Ipsilateral radiotherapy was performed on 37 patients (46.8%), bilateral neck plus mucosal irradiation was performed in 42 patients (53.2%). Ten patients (12.2%) developed a primary tumor during the follow-up. The actuarial survival rates of all patients 2, 5 and 10 years after diagnosis were 50.9, 25.3 and 18.5%, respectively. Patients with nodal stage N2b, N2c and N3 had a significantly poorer prognosis than those with nodal stage N1 and N2a (p = 0.0239). The survival in patients with metastatic nodes in the supraclavicular region (level IV) was significantly poorer than that of patients with involvement of the upper-middle jugular lymph nodes (p = 0.0003). We observed a statistically significant better survival in patients receiving bilateral neck plus mucosal irradiation (p = 0.0003). CONCLUSIONS: Initial N-category and metastasis localization were the most important prognostic factors and nodal relapse the major cause of treatment failure, thus optimal management of cervical nodes appears crucial for the success of treatment. Patients receiving bilateral neck plus mucosal irradiation had a higher survival rate than those who received ipsilateral irradiation. PMID- 16479146 TI - Practical use of MESNA in atelectatic ears and adhesive otitis media. AB - OBJECTIVE: MESNA is a synthetic sulfur compound that produces mucolysis by disrupting disulfide bonds. This study aimed to address indications and technique of MESNA application in otologic surgery, especially in retraction pockets and adhesive otitis media. METHODS: MESNA application was performed in 42 ears of 39 patients. The diagnoses were retraction pockets fixed to incudostapedial joint, stapes or promontorium, and adhesive otitis media in 24 and 17 ears, respectively. Calculations were performed according to 24 ears of 23 patients with follow-up data. RESULTS: One or more of the following surgical interventions were performed: MESNA application alone and ventilation tube insertion, reinforcement or mastoidectomy in addition to MESNA application. Sensorineural hearing loss was not encountered after MESNA application. The operation was successful in 79.2%. Overall revision surgery was needed in 20.8% of the ears. In 71.4% of the ears treated with MESNA alone, revision surgery was needed. CONCLUSION: In the light of our experience, we advocate the use of MESNA in atelectatic ears, because it makes the operation easy and safe by allowing elevation of the tympanic membrane by its mechanical and chemical actions. PMID- 16479147 TI - First findings concerning airflow in noses with septal deviation and compensatory turbinate hypertrophy--a model study. AB - BACKGROUND: The nasal septal deviation is a common cause of nasal obstruction. On the other hand, many septal deviations are asymptomatic. It seems a physiological adaptation occurs on both sides. Septal deviation leads to internal nasal asymmetry, which in turn causes compensatory change in turbinate morphology (e.g. turbinate hypertrophy respectively hypotrophy). This mechanism is investigated with the help of fluid dynamic experiments and functional rhinologic diagnostics. METHODS: Functional models of the nose (modified Mink's boxes) were used and assessment was made by acoustic rhinometry and rhinoresistometry, followed by flow dynamic investigations. Septal deviations of varying position, together with turbinates of differing grades of hypertrophy, were simulated and assessed. RESULTS AND CONCLUSIONS: We observed in models of septal deviation an increase in flow resistance on the ipsilateral side as a result of friction of flow particles in the narrowing. Furthermore, on the opposite side of the deviation, the enlargement of the stream channel did not generally lead to a reduction in flow resistance, but rather to a 'dead space', where only a slow-circling eddy was observed. This eddy causes an increase in turbulence. In vivo turbinate hypertrophy occurs to fill this dead space, thereby reducing turbulent flow without a significant increase in resistance. In cases of moderate septal deviation, compensatory mechanisms of the turbinates can lead to a normalization of nasal airflow and surgical therapy would not be indicated. Deviations in the anterior part of the septum seem to be more symptomatic, because the mechanism is missing and due to the physiological narrowing of the nasal isthmus. To differ between physiologic and pathologic deviation, functional diagnostics are needed. PMID- 16479148 TI - Antiproliferative effect of isosteviol on angiotensin-II-treated rat aortic smooth muscle cells. AB - Isosteviol is a derivative of stevioside, a constituent of Stevia rebaudiana, which is commonly used as a noncaloric sugar substitute in Japan and Brazil. The aims of this study were to examine whether isosteviol alters angiotensin-II induced cell proliferation in rat aortic smooth muscle cells. Cultured rat aortic smooth muscle cells were preincubated with isosteviol, then stimulated with angiotensin II, after which [(3)H]thymidine incorporation and endothelin-1 secretion were examined. Isosteviol (1-100 micromol/l) inhibits angiotensin-II induced DNA synthesis and endothelin-1 secretion. Measurements of 2'7' dichlorofluorescin diacetate, a redox-sensitive fluorescent dye, showed an isosteviol-mediated inhibition of intracellular reactive oxygen species generated by the effects of angiotensin II. The inductive properties of angiotensin II on extracellular signal-regulated kinase (ERK) phosphorylation were found reversed with isosteviol and antioxidants such as N-acetylcysteine. In summary, we speculate that isosteviol inhibits angiotensin-II-induced cell proliferation and endothelin-1 secretion via attenuation of reactive oxygen species generation. Thus, this study provides important insights that may contribute to the effects of isosteviol on the cardiovascular system. PMID- 16479149 TI - Involvement of voltage-dependent Ca(2+) channel beta(3) subunit in the autonomic control of heart rate variability. AB - Noradrenaline release from sympathetic nerve terminals is dependent on Ca(2+) entry through neuronal voltage-gated N-type Ca(2+) channels. The accessory beta(3) subunits of Ca(2+) channels (Ca(V)beta(3)) are preferentially associated with the alpha(1B) subunit to form N-type Ca(2+) channels, and are therefore expected to play a functional role in the stimulation-evoked release of noradrenaline. In this study, we employed Ca(V)beta(3)-null, Ca(V)beta(3) overexpressing (Ca(V)beta(3)-Tg), and wild-type (WT) mice to investigate the possible roles of Ca(V)beta(3) in the sympathetic regulation of heart rate in vivo. Telemetry was used to monitor the ECG and both time and frequency domain analyses were carried out to evaluate heart rate variability. In the frequency domain analysis, power spectral density of the RR interval series was computed using the fast Fourier transform algorithm. The resting heart rate was increased in Ca(V)beta(3)-Tg mice compared with both Ca(V)beta(3)-null and WT mice. Mice overexpressing Ca(V)beta(3) displayed decreased heart rate variability, which was measured by the time domain analysis of the standard deviation of RR intervals. In the frequency domain analysis, Ca(V)beta(3)-Tg mice showed decreased spectral powers compared with WT and Ca(V)beta(3)-null mice. Pharmacological blockade of beta-adrenergic receptors with metoprolol decreased the heart rate in all genotypes, but the extent of the decrease was most obvious in Ca(V)beta(3)-Tg mice. On the other hand, the spectral powers were decreased in response to parasympathetic blockade (atropine) in WT and Ca(V)beta(3)-Tg mice. These results indicate the functional roles of Ca(V)beta(3) in regulating sympathetic nerve signaling. PMID- 16479150 TI - Building a cathedral. PMID- 16479151 TI - Substrates of the BRCA1-dependent ubiquitin ligase. AB - Discovering the precise function of the breast and ovarian specific tumor suppressor, BRCA1, has proven to be quite complicated. It has been determined that BRCA1, together with BARD1, comprise an E3 ubiquitin ligase. Since it is now known that BRCA1 is an enzyme, the challenge for BRCA1 research is to learn how this enzymatic activity functions in normal breast and ovarian cells in order to suppress cancerous transformation. This review will survey the known ubiquitination substrates of BRCA1 and suggest how these reactions may influence the genomic stability and proliferation of breast cells. PMID- 16479152 TI - Biological therapies: twenty-five years of progress in cancer therapy. PMID- 16479153 TI - Is it easy to stop RNA polymerase? AB - Among transcription factors that bind to bacterial RNA polymerase (RNAP) and modulate its activity, a number of small molecules irreversibly inhibit RNAP thereby causing cell death. To be of clinical significance such inhibitors must (1) inhibit a broad range of bacterial RNAPs but not affect human cells, (2) penetrate bacterial cell walls and (3) circumvent bacterial resistance mechanisms. Rifamycins, the only class of RNAP inhibitors that have found their way into clinical practice, are widely used in the treatment of tuberculosis and leprosy. However, the practical value of this class of antibiotics is limited by a rapid rise in resistant bacterial isolates. In this review we focus on recent advances in studies of prokaryotic transcription that allow a detailed structural and functional characterization of a number of RNAP/rifamycins complexes, thereby opening new opportunities for the design of superior antibacterial agents. PMID- 16479155 TI - Decatenation checkpoint deficiency destabilizes the stem cell genome. PMID- 16479154 TI - Cyclin D1 overexpression increases the susceptibility of human U266 myeloma cells to CDK inhibitors through a process involving p130-, p107- and E2F-dependent S phase entry. AB - Dysregulation of cyclin D1 expression is one of the most common genetic aberrations found in hematopoietic malignancies, including multiple myeloma. To address the effects of cyclin D1 overexpression might have on the response of malignant hematopoietic cells to CDK inhibitors, the impact of ectopic cyclin D1 overexpression on the response of human multiple myeloma U266 cells to various cyclin-dependent kinase (CDK) inhibitors was examined. Cyclin D1 overexpression markedly increased the apoptotic response of cells to the CDK inhibitors flavopiridol, roscovitine, and R-roscovitine. Ectopic expression of cyclin D1 resulted in p21(CIP1) accumulation, an effect that was diminished by CDK inhibitor exposure. In pRb-null U266 cells, enforced overexpression of cyclin D1 diminished CDK inhibitor-mediated dephosphorylation of the pocket proteins p130 and p107, reduced binding of E2F1 and E2F4 to p130 and p107, and attenuated inhibition of E2F activity. Notably, CDK inhibitors failed to reduce the S phase fraction in cyclin D1/U266 cells in contrast to effects in their wild-type counterparts. Finally, cyclin D1/U266 cells exhibited diminished basal NF-kappaB activity compared to controls, which was essentially completely abrogated by CDK inhibitor exposure. Together, these findings suggest that dysregulation of cyclin D1 sensitizes human myeloma cells to the actions of CDK inhibitors through mechanisms involving interference with p21(CIP1) expression, dephosphorylation of pocket proteins and inactivation of E2Fs culminating in S phase entry, as well as inactivation of NF-kappaB, leading to apoptosis rather than growth arrest. PMID- 16479156 TI - TFII-I: connecting mitogenic signals to cell cycle regulation. AB - Reproduction, or the faithful passage of genetic information from one cell to its progeny, is central to life. To achieve reproduction faithfully cells have developed the cell cycle during which error-free replication of DNA, followed by division of the nucleus and partitioning of the cytoplasm yields two daughter cells. Prior to committing to reproduction, cells must sense mitogen levels in their environment and transduce those signals to the nucleus through a series of biochemical steps, resulting in spatial and/or temporal activation of genes that will drive cell cycle progression past the restriction point and initiate DNA replication. One way external signals are transmitted to the nucleus is via mitogen inducible transcription factors that shuttle between the cytoplasm and nucleus. Here we introduce a newly discovered pathway by which TFII-I, a growth signal induced transcription factor, operates in the prerestriction point and mitogen dependent phase of the cell cycle. We also discuss a potential role for TFII-I in DNA repair and how it might be involved in signal dependent DNA repair versus cell cycle. PMID- 16479157 TI - Chemokine signaling regulates apoptosis as well as immune cell traffic in host defense. AB - In the struggle for optimal host defense against infection with viruses, two major events are critical: death of the infected host cell and proper immune cell activation at the site of infection. Here we summarize our recent work indicating that chemokines exhibit a distinct capacity to regulate both of these events. We put particular emphasis on a recently completed study indicating that chemokine CCL5 may prevent cell death and thereby preserve innate immune cell function in the setting of viral infection. In addition, we introduce new work to support the more traditional role of CCL5 in mediating adaptive immune cell traffic and activation in this same setting. PMID- 16479158 TI - Pocket proteins control white versus brown fat cell differentiation. PMID- 16479159 TI - The mechanism of hematopoietic progenitor cell immortalization by MLL-ENL. AB - The t(11;19) translocation gives rise to the MLL-ENL fusion protein and is frequently found in infant myeloid and lymphoid leukemias. Immortalized myeloid cell lines can be generated by expression of MLL-ENL in murine hematopoietic progenitors. By establishing myeloid cell lines with conditional expression of MLL-ENL, we recently demonstrated that MLL-ENL is necessary to maintain immortalization and sustain the expression of a characteristic pattern of Hox genes. The cell lines can be induced to undergo terminal differentiation by inhibition of MLL-ENL expression or by treatment with G-CSF. Expression of Hoxa genes is reduced in cells differentiating as a result of MLL-ENL loss, but is maintained in G-CSF treated cells. Thus, although aberrant maintenance of Hoxa gene expression may play an important role in MLL-ENL induced leukemia, the contribution of this pathway to immortalization is critically dependent on the cytokine environment of the immortalized myeloid cells. PMID- 16479160 TI - Preventing fatal destruction: inhibitors of the anaphase-promoting complex in meiosis. AB - The anaphase-promoting complex/cyclosome (APC/C) is a multi-subunit ubiquitin ligase whose major functions in the cell cycle are the initiation of sister chromatid separation and the inactivation of cyclin-dependent kinases. This complex is also essential for meiosis, a specialized form of the cell cycle characterized by two consecutive rounds of chromosome segregation. To ensure a proper meiotic cell cycle, the activity of APC/C needs to be tightly controlled. It is now evident that inhibitors of APC/C play pivotal roles to avert its untimely activation. During prophase I, this ubiquitin-ligase must be kept inactive to prevent precocious sister chromatid separation. Studies in yeast showed that this inhibition is mediated by a specific subunit of the complex. Accurate chromosome segregation in meiosis I depends on spindle checkpoint proteins such as Mad2 which delay APC/C activation in response to an erroneous spindle attachment of chromosomes. Additional APC/C antagonists are known to block complete cyclin destruction between meiosis I and II, thereby ensuring that cyclin dependent kinases remain active and that DNA replication does not occur. Inhibitors of APC/C also mediate the cytostatic factor induced metaphase II arrest of oocytes. This review highlights the current knowledge about the role and relevance of these diverse regulators of the meiotic APC/C. PMID- 16479161 TI - Securin associates with APCCdh1 in prometaphase but its destruction is delayed by Rae1 and Nup98 until the metaphase/anaphase transition. AB - Precisely timed ubiquitin-mediated proteolysis of mitotic regulators by the anaphase-promoting complex (APC) governs the orderly passage of cells through mitosis. The established view is that Cdc20-activated APC (APC(Cdc20)) mediates the destruction of cyclin B and securin at the metaphase/anaphase transition, and that Cdh1-activated APC (APC(Cdh1)) has no role in this process. We recently reported that securin, but not cyclin B, is prematurely targeted for destruction by the APC in mutant mice that have low levels of the nuclear transport factors Rae1 and Nup98. We found that Rae1 and Nup98 assemble into a complex with APC(Cdh1) in prometaphase and act to delay APC(Cdh1)-mediated ubiquitination of securin until the metaphase/anaphase transition. Here we show that Rae1 and Nup98 not only form a complex with APC(Cdh1) in prometaphase but also with securin. This finding suggests that the Rae1-Nup98 complex does not inhibit early destruction of securin by preventing APC(Cdh1) from binding to securin, but by preventing ubiquitination of APC(Cdh1)-bound securin. We propose that the formation of APC(Cdh1)-securin complexes in prometaphase primes the cell for rapid securin degradation after release of the inhibitory Rae1-Nup98 complex at the metaphase/anaphase transition. We further report here that mutant mice with low levels of the Rae1-Nup98 complex are not prone to develop spontaneous tumors, despite massive aneuploidy. However, Rae1/Nup98 mutant mice are significantly more susceptible to DMBA-induced lung tumors than wild-type mice, indicating that combined Rae1/ Nup98 haplo-insufficiency does promote tumorigenesis when certain cancer-critical genes are also mutated. PMID- 16479162 TI - Diverse epigenetic profile of novel human embryonic stem cell lines. AB - Human embryonic stem cells (hESCs) are a promising model for studying mechanisms of regulation of early development and differentiation. OCT4, NANOG, OCT4-related genes and some others were recently described to be important in pluripotency maintenance. Lesser is known about molecular mechanisms involved in their regulation. Apart from genetic regulation of gene expression epigenetic events, particularly methylation, play an important role in early development. Using RT PCR we studied the expression of pluripotency-related genes OCT4, NANOG, DPPA3 and DPPA5 during hESCs differentiation to embryoid bodies. Analysis of methylation profiles of promoter or putative regulatory regions of the indicated genes demonstrated that expression of the pluripotency-maintaining genes correlated with their methylation status, whereas methylation of DPPA3 and DPPA5 varied between cell lines. We propose that DNA methylation underlies the developmental stage-specific mechanisms of pluripotency-related genes expression and reactivation and may have an impact on differentiation potential of hESC lines. PMID- 16479163 TI - Direct interaction of p21 with p50, the small subunit of human DNA polymerase delta. AB - Using a yeast two-hybrid screening technique and the p50 subunit of human DNA polymerase delta (pol delta) as a bait, p21 was found to interact with the p50 subunit of pol delta. A direct interaction between p21 and p50 was confirmed by using ELISA and pull-down assays with purified proteins. The interaction sites between p50 and p21 were mapped by pull down assays with GST deletion mutants. Residues 127-193 constitute the primary interaction region on p50 to which p21 binds, while p50 binds to the C-terminal 26 residues of p21. A histone kinase activity was associated with the highly purified calf thymus pol delta and addition of purified recombinant p21 inhibited the kinase activity in a dose dependent manner. p50 is phosphorylated in vivo and can be phosphorylated by CDK2/cyclinA in vitro. In vivo evidence of p21 association with p50 was obtained by coimmunoprecipitation using MCF7 cells. It was also shown that the association of p21 with p50 and other components of the pol delta complex increased in MCF7 cells treated with adriamycin. Our results suggested that p50 might target or anchor p21 to pol delta complex upon certain DNA damage such as adriamycin treatment. PMID- 16479165 TI - Prostate cancer prevention through pomegranate fruit. AB - Prostate cancer (CaP) is the second leading cause of cancer-related deaths among U.S. males with a similar trend in many Western countries. CaP is an ideal candidate disease for chemoprevention because it is typically diagnosed in men over 50 years of age, and thus even a modest delay in disease progression achieved through pharmacological or nutritional intervention could significantly impact the quality of life of these patients. In this regard we and others have proposed the use of dietary antioxidants as candidate CaP chemopreventive agents. The fruit pomegranate derived from the tree Punica granatum has been shown to possess strong antioxidant and anti-inflammatory properties. In a recent study, we showed that pomegranate fruit extract (PFE), through modulations in the cyclin kinase inhibitor-cyclin-dependent kinase machinery, resulted in inhibition of cell growth followed by apoptosis of highly aggressive human prostate carcinoma PC3 cells. These events were associated with alterations in the levels of Bax and Bcl-2 shifting the Bax:Bcl-2 ratio in favor of apoptosis. Further, we showed that oral administration of a human acceptable dose of PFE to athymic nude mice implanted with CWR22Rnu1 cells resulted in significant inhibition of tumor growth with concomitant reduction in secretion of prostate-specific antigen (PSA) in the serum. The outcome of this study could have a direct practical implication and translational relevance to CaP patients, because it suggests that pomegranate consumption may retard CaP progression, which may prolong the survival and quality of life of the patients. PMID- 16479164 TI - Identification of tumor precursor cells in the brains of primates with radiation induced de novo glioblastoma multiforme. AB - The pathogenesis of de novo glioblastoma multiforme (GBM) is poorly understood and precursor cells are not known. To gain insight into the pathogenesis of GBM we analyzed brains from primates that developed de novo tumors ten years after whole brain radiation. Four animals had clinical and radiological evidence of GBM, and two animals had no evidence of GBM at the time of euthanization. Tumor precursor cells were identified diffusely scattered in the grossly normal white matter of all animals including two monkeys without evidence of GBM by MR-imaging or on autopsy examination. Tumor precursors demonstrated cellular atypia and mitoses, and were negative for tumor-associated markers GFAP, EGFR and p53. The cells were positive for Ki67 and N-CoR, the nuclear corepressor of astroglial differentiation. These results suggest that radiation-induced nuclear damage to neural stem cells or early astrocytic precursor cells can prevent normal differentiation and lead to tumor development. The findings provide insight into the tumorigenesis of de novo GBMs and suggest a new strategy for treatment of these lethal tumors by targeting both inactivation of N-CoR and inhibition of EGFR. PMID- 16479166 TI - A potential molecular link between aerobic glycolysis and cancer. PMID- 16479168 TI - Alternative splicing increases complexity of stem cell transcriptome. AB - Development of highly anticipated stem cell-based therapies requires a detailed understanding of mechanisms regulating biological properties of these cells. Comprehensive identification of all biological molecules produced in stem cells is an important step toward this goal. During the past several years, microarray studies have essentially identified genes that are transcriptionally activated in various embryonic and adult stem cell populations. However, the extent of post transcriptional modifications within the stem cell transcriptome remained undetermined. Recently, we presented evidence that thousands of genes expressed in hematopoietic and embryonic stem cells undergo alternative splicing. Using combined computational and experimental analyses, we found that the frequency of alternative splicing is especially high in tissue-specific genes, as compared to ubiquitous genes. Our results also indicate that negative regulation of constitutively active splicing sites can be a prevalent mode for generation of splicing variants, and that alternative splicing is generally not conserved between orthologous genes in human and mouse. Here, we discuss the implications of our findings for stem cell biology, and present possible approaches toward genome-wide identification and characterization of splice variants. PMID- 16479167 TI - The GOD of hematopoietic stem cells: a clonal diversity model of the stem cell compartment. AB - Hematopoietic stem cells (HSC) show heterogeneous behavior even when isolated as phenotypically homogeneous populations. The cellular and molecular mechanisms that control the generation of diversity (GOD) in the HSC compartment are not well understood, but have been the focus of much debate. There is increasing evidence that the most important HSC functions, self-renewal and differentiation, are epigenetically preprogrammed and therefore predictable. Indeed, recent data show that the adult HSC compartment consists of a limited number of functionally distinct subsets of HSC. This contradicts older models of HSC behavior, which postulated a single type of HSC that can be continuously molded into different subtypes of HSC. We propose a clonal diversity model where the adult HSC compartment consists of a fixed number of different types of HSC, each with epigenetically preprogrammed behavior. Aging or disease may change the overall function of the HSC population. The model predicts that these changes reflect the relative composition of the HSC subsets, rather than changes in individual HSC. This view has implications for using HSC in experimental and clinical settings. Selection for the appropriate subsets of HSC, rather than attempts to force HSC to adjust, should improve their utility in transplantation and gene transfer applications. PMID- 16479170 TI - Grasses like mammals? Redundancy and compensatory regulation within the retinoblastoma protein family. AB - The retinoblastoma (RB) protein family plays a conserved and inhibitory role in cell cycle progression in higher eukaryotes. In mammals, this family includes, in addition to RB, the related (RBR) proteins p107 and p130, which appear to have both specific and redundant functions compared to those of the prototypical RB protein. Whereas most plant species seem to possess only one RBR gene, a recent study has shown that in maize there are two types of distinctly regulated RBR proteins, RBR1 and RBR3. Expression of RBR3 RNA is controlled by the RBR1-E2F pathway, and it is upregulated upon inhibition of RBR1 activity by the wheat dwarf virus RepA protein in tissue culture, indicating the presence of a specific compensatory mechanism sustaining high pocket protein activity. Database mining and phylogenetic analyses suggest the presence of two distinct RBR genes to be a unique feature of grasses among plants, which might help to explain their recalcitrance to genetic transformation. PMID- 16479171 TI - Geminin's double life: chromatin connections that regulate transcription at the transition from proliferation to differentiation. AB - During embryonic development, transitions between cellular programs regulating progenitor cell proliferation and differentiation must be precisely coordinated and temporally controlled to ensure that a proper number of cells are allocated to various structures. The novel coiled-coil protein Geminin was previously characterized as a dual function molecule with roles both in maintenance of genome integrity through regulation of DNA replication licensing and in control of neural cell fate during embryonic development. However, the mechanistic basis of Geminin's activities during embryogenesis and the connections to its cell cycle regulatory role were unknown. Recently, some of Geminin's activities in regulating transcription were shown to occur through interactions with Brg1, the catalytic subunit of the SWI/SNF chromatin-remodeling complex. During development of the nervous system, Geminin controls the transition from proliferating precursor to differentiated post-mitotic neuron by modulating interactions between SWI/SNF and bHLH transcription factors that are critical for neurogenesis. In other developmental contexts, Geminin mediates proliferative differentiative transitions through interactions with Six3 nd Hox transcription factors and Polycomb Group proteins. Interactions of Geminin with Polycomb and SWI/SNF complex proteins link its transcriptional activities to modulation of chromatin structure. Here we incorporate recent findings regarding Geminin's regulatory roles in coordinating proliferation and differentiation during embryogenesis. PMID- 16479172 TI - The CDK inhibitor p18Ink4c is a tumor suppressor in medulloblastoma. AB - Medulloblastoma (MB) is the most common malignant pediatric brain tumor which is thought to originate from cerebellar granule cell precursors (CGNPs) that fail to properly exit the cell cycle and differentiate. Although mutations in the Sonic Hedgehog (Shh) signaling pathway occur in 30% of cases, genetic alterations that account for MB formation in most patients have not yet been identified. We recently determined that the cyclin D-dependent kinase inhibitor, p18(Ink4c), is expressed as CGNPs exit the cell cycle, suggesting that this protein might play a central role in arresting the proliferation of these cells and in timing their subsequent migration and differentiation. In mice, disruption of Ink4c collaborates independently with loss of p53 or with inactivation of the gene (Ptc1) encoding the Shh receptor, Patched, to induce MB formation. Whereas loss of both Ink4c alleles is required for MB formation in a p53-null background, Ink4c is haplo-insufficient for tumor suppression in a Ptc(1+/-) background. Moreover, MBs derived from Ptc(1+/-) mice that lack one or two Ink4c alleles retain wild-type p53. Methylation of the INK4C (CDKN2C) promoter and complete loss of p18(INK4C) protein expression were detected in a significant fraction of human MBs again pointing toward a role for INK4C in suppression of MB formation. PMID- 16479173 TI - AKT pathway is activated by 1, 25-dihydroxyvitamin D3 and participates in its anti-apoptotic effect and cell cycle control in differentiating HL60 cells. AB - Differentiation therapy for cancer is a developing treatment modality that is based on the anti-proliferative effects associated with differentiation of the malignant cells. 1,25-dihydroxyvitamin D(3) (1,25D) and its analogs are currently being evaluated clinically, alone or in combination with other agents, for treatment of several neoplastic diseases, but their usefulness as single agents may be limited by the enhancement of cell survival in some cell types exposed to 1,25D. In this study we evaluated the role of AKT signaling pathway, known to be anti-apoptotic in diverse cell types, in enhancing the survival of human leukemia HL60 cells induced to differentiate with 1,25D. We found that the phosphorylation and activity of AKT, as well as of its downstream targets, are increased after the exposure to 1,25D. Treatment of HL60 cells with PI3K inhibitors LY294002 and Wortmannin, which decrease the activity of the AKT pathway, induced apoptosis, but this effect was reduced in cells simultaneously treated with 1,25D. Interestingly, LY294002 and Wortmannin also accentuated the 1,25D-induced G(1) to S phase cell cycle block in HL60 cells, and this was associated with an increased expression of p27Kip1. Thus, a combination of 1,25D with inhibitors of AKT pathway is strongly anti-proliferative and should therefore be considered for differentiation therapy of myeloid leukemia. PMID- 16479174 TI - Microcephalin encodes a centrosomal protein. PMID- 16479169 TI - HEF1-aurora A interactions: points of dialog between the cell cycle and cell attachment signaling networks. AB - Regulated timing of cell division cycles, and geometrical precision in the planar orientation of cell division, are critical during organismal development and remain important for the maintenance of polarized structures in adults. Mounting evidence suggests that these processes are coordinated at the centrosome through the action of proteins that mediate both cell cycle and cell attachment. Our recent work identifying HEF1 as an activator of the Aurora A kinase suggests a novel hub for such integrated signaling. We suggest that defects in components of the machinery specifying the temporal and spatial integration of cell division may induce cancer and other diseases through pleiotropic effects on cell migration, proliferation, apoptosis, and genomic stability. PMID- 16479175 TI - Lessons learned fighting HIV can be applied to anti-cancer drug design. PMID- 16479176 TI - Highlights of the Twelfth Annual COSEHC National Scientific Session. PMID- 16479178 TI - Relationships between heart rate variability and urinary albumin excretion in patients with type 2 diabetes. AB - BACKGROUND: Although in type 1 diabetes the close association between heart rate variability and urinary albumin excretion (UAE) is recognized even in patients with normoalbuminuria, this association has not yet been fully established in patients with type 2 diabetes. Therefore, we investigated the association in patients with type 2 diabetes. PATIENTS AND METHODS: All the hospital's 185 inpatients with type 2 diabetes were prospectively enrolled. Heart rate variability was evaluated by coefficients of variance of RR intervals (CVRR). RESULTS: The mean age, duration of diabetes, and hemoglobin A1C of the patients were 59.7+/-9.9 years, 10.4+/-7.8 years, and 9.7+/-2.3%, respectively. An analysis of the patients showed a significant negative correlation between CVRR and log10-transformed (log) UAE (R=-0.3340, P <0.0001). CVRR showed a significant negative correlation with age, duration of diabetes, hemoglobin AIC, systolic blood pressure, diastolic blood pressure, and triglyceride level. Log UAE showed a significant positive correlation with body mass index, hemoglobin A1C, systolic blood pressure, diastolic blood pressure, total cholesterol, and triglyceride level. In the macroalbuminuric group (UAE above 300 mg/g creatinine; n=57), although CVRR showed a significant negative correlation with log UAE (R=-0.3571, P= 0.0064), but in normoalbuminuric (UAE below 30 mg/g Cr; n=79) and in microalbuminuric groups (30 to 300 mg/g Cr; n = 49), CVRR and log UAE showed no correlation. CONCLUSIONS: Our data suggest that in type 2 diabetes, the association between CVRR and UAE is significant only in patients with macroalbuminuria. PMID- 16479177 TI - Effects of riboflavin and folic acid supplementation on plasma homocysteine levels in healthy subjects. AB - BACKGROUND: Observational studies have shown an inverse relationship between vitamin B2 status and total homocysteine levels, a risk factor for cardiovascular disease. We hypothesize that intervention with riboflavin will lower total homocysteine levels. The total homocysteine lowering by the three genotypes (CC, CT, TT) of methylenetetrahydrofolate reductase polymorphism (677C-->T) was also studied. METHODS: The decrease in total homocysteine levels after supplementation with riboflavin (10 mg/d) or folic acid (1 mg/d) for 3 weeks was compared in two groups of healthy subjects (17 per group, matched by age and gender) (Phase 1). Then, both groups received supplementation with folic acid and riboflavin for an additional 3 weeks (Phase 2). RESULTS: During Phase 1, total homocysteine levels were lowered by 2% or 4% after supplementation with riboflavin or folic [corrected] acid, respectively, although neither decrease was statistically significant (P=0.50 and 0.19). Compared to subjects of CC genotype, total homocysteine lowering in subjects of CT genotype was approaching significance (P=0.059) for the folic acid group, but not for the riboflavin group. After Phase 2, total homocysteine levels were not lowered significantly in either the folic acid (1%) or the riboflavin (2%) group. However, in the folic acid-riboflavin combined group, total homocysteine lowering in subjects of TT type was larger when compared to subjects of CC and CT types (P=0.007). CONCLUSIONS: Riboflavin supplementation did not lower total homocysteine levels in healthy subjects with CC type of C677T polymorphism. However, supplementation with folic acid or with both folic acid and riboflavin may be important for CT and TT subjects in optimizing their homocysteine metabolism. These findings are relevant in characterizing the factors controlling the high total homocysteine levels for subjects of CT and TT genotypes. PMID- 16479179 TI - Orbital lymphomas: a clinicopathologic study of a rare disease. AB - OBJECTIVE: To evaluate the clinicopathologic features and prognosis of patients with orbital lymphomas. METHODS: Clinical and pathologic data of 35 patients with biopsy-proven orbital lymphoma diagnosed at a tertiary care hospital from 1992 to 2001 were reviewed. Lymphomas were divided into low-grade and high-grade lymphomas. Survival of patients was compared according to age, gender, disease site, extent of disease, tumor grade, and treatment modality by using log rank test. RESULTS: Median patient age was 75 years (23-94) and the male-to-female ratio was 1:2.9. Twenty-three patients (66%) were diagnosed with low-grade lymphoma, and 12 patients (34%) were found to have high-grade lymphoma. Among low grade lymphomas, marginal zone lymphoma (n=6), follicle center cell lymphoma (n= 6), and small lymphocytic lymphoma (n=5) were common entities, whereas diffuse large cell B-cell lymphoma (n=5) was the most common entity in patients with high grade lymphoma. Disease was clinically localized in 74% of patients at the time of diagnosis. Radiation alone or with chemotherapy was the primary treatment modality in 83% of patients. All except one patient had an objective response to therapy. Over the median follow-up period of 47 months (range, 1.5-141 months), disease recurred in 37% patients who achieved a complete response. The estimated 5- and 10-year survival rates were 64% and 42%, respectively. Overall, 13 (37%) patients died, 6 with high-grade and 7 with low-grade lymphoma. No clinical variable was found to be prognostically significant with respect to survival. CONCLUSIONS: Orbital lymphoma is a disease of the elderly with a female preponderance. It tends to be localized to the orbit at the time of diagnosis and responds well to local or systemic therapy. PMID- 16479180 TI - Staphylococcus aureus endocarditis: The Grady Memorial Hospital Experience. AB - BACKGROUND: Staphylococcus aureus is a common cause of endocarditis. Previous studies have shown that the mortality rate associated with S aureus endocarditis remains high, and methicillin resistance is becoming more prevalent. METHODS: Over the past 4 years, 283 patients with suspected endocarditis were referred to the cardiology department of Grady Memorial Hospital for echocardiography. Their clinical course and demographic information was entered into a database. Among these patients, 87 (30.6%) were confirmed to have endocarditis, with 41 (47.1%) of them culture positive for either methicillin-resistant (MRSA) or methicillin sensitive S aureus (MSSA). Of these patients, 14 were MRSA culture positive, and 27 were MSSA culture positive. These two groups were compared with respect to demographic information and morbidity and mortality outcomes. RESULTS: The MRSA and MSSA groups were found to be similar upon analysis in all baseline characteristics. There was a non-statistical trend among the two groups in origin of infection, as the MRSA group tended toward nosocomial infection more often. Outcomes, including surgery, death, and complications were also found to be similar between the two groups. CONCLUSIONS: There does not seem to be difference in the outcome of MRSA versus MSSA endocarditis. Nosocomial infection may predispose patients to MRSA endocarditis, but a larger study with a greater sample size is necessary to evaluate these questions more accurately. PMID- 16479181 TI - The necessity of culture for the diagnosis of tinea pedis. AB - BACKGROUND: This study examined the consistency between the clinical diagnosis of tinea pedis and the results of direct fungal examination, prepared with 10% potassium hydroxide, and culture. METHODS: 2,427 patients clinically diagnosed with tinea pedis who presented to the mycology laboratory were reviewed retrospectively for the outcomes of direct fungal examination and culture. RESULTS: Direct examination was positive in 54.3% and culture was positive in 36.6% of the cases. The sensitivity and specificity of direct microscopy were 95.7% and 69.6%, respectively CONCLUSIONS: The clinical diagnosis of tinea pedis can be misleading, since it features lesions that can also be present in some other skin diseases and direct microscopy may be insufficient to confirm the diagnosis. Therefore, we suggest using culture for a definitive diagnosis. PMID- 16479183 TI - Two-dimensional echocardiography for the diagnosis of interventricular septum perforation by a temporary pacing catheter. AB - This case study describes a patient with complete heart block inadvertently paced from the left ventricular posterior wall due to perforation of interventricular septum by a temporary pacing catheter. This is a rare but potentially fatal complication of pacing. The frontal chest radiograph neither identified abnormalities nor could determine the exact site of the catheter tip. The electrocardiogram revealed a right bundle branch block pattern. Echocardiography was performed immediately at bedside and the diagnosis was made. The temporary pacing catheter was removed without complications and, the next day, the patient underwent permanent pacemaker implantation. Given its noninvasiveness and availability, echocardiography is a highly effective means of assessing pacemaker complications such as catheter perforation. PMID- 16479182 TI - Cardiac papillary fibroelastoma presents as an acute embolic stroke in a 35-year old African American male. AB - This is an interesting case of a young patient suffering an acute embolic stroke in the middle cerebral artery distribution, who was later found to have a papillary fibroelastoma on the mitral valve. The mass was first recognized by transesophageal echocardiography and eventually resected surgically. The retrieved specimen had classic histologic findings of a papillary fibroelastoma. A thrombus was noted on the tip of the specimen, supporting the theory that these masses are risks for strokes secondary to damage along the endothelial lining predisposing to subsequent fibrin deposition and mural thrombus formation. PMID- 16479184 TI - A thyroidectomy case with a surprising pathologic result: three different birds in one nest. AB - Neoplasms of the thyroid include types that range from indolent, localized papillary carcinomas to lethal anaplastic disease. Hurthle cell neoplasm is a histopathologic subtype that signals poor prognosis. Herein, we present the case of a 72-year-old female patient with multinodular goiter who, although fine needle aspiration biopsy revealed a benign result, underwent thyroid surgery because of the diameter of the largest nodule. Histopathologically, the neoplasm was diagnosed as papillary thyroid microcarcinoma on one lobe and Hurthle cell neoplasm on the other, whereas the remainder of the thyroid showed a typical pattern of colloidal goiter, all of which are not commonly encountered in one patient. Although multinodular goiter is usually acknowledged as a sign of a benign process, its risk of malignancy is no less than that of a solitary thyroid nodule. The age of the patient, the duration of the disease, or the number of the nodules should not mislead the physician, nor affect the therapeutic approach. PMID- 16479185 TI - Heart failure in rupture of a sinus of valsalva aneurysm. AB - We report a 22-year-old man who developed shortness of breath after lifting weights and then developed acute heart failure due to rupture of an aneurysm of the right sinus of Valsalva into the right ventricle. The patient developed dyspnea, and clinical findings included tachycardia, wide pulse pressure, bounding carotid and peripheral pulses, pulmonary crackles, and prominent continuous precordial murmur with thrill. Transesophageal echocardiogram with Doppler examination confirmed the diagnosis. The patient underwent surgery with cardioplegia directly infused into the coronary arteries with excision of redundant tissue and closure of the defect with a Dacron patch. He has been asymptomatic since surgery. This condition must to be included in the differential diagnosis for young patients with heart failure. PMID- 16479186 TI - Myxedema ascites with elevated serum CA 125 concentration. AB - We report a case of myxedema ascites and markedly elevated serum CA 125 concentration. The cause of ascites and elevated tumor markers in hypothyroidism remains unknown. Diagnosis was characterized by no evidence of malignancy seen by transvaginal ultrasonography or abdominal computed tomography and ascites resolution with serum CA 125 normalization after adequate hormonal treatment. Our data suggest that hypothyroidism should be considered in patients with ascites and elevated serum CA 125. PMID- 16479187 TI - Correction of renal hypertension after kidney transplantation from a donor with Gitelman syndrome. AB - The relationship between the kidney and blood pressure control is complex. Monogenetic forms of hypertension have recently been identified that implicate specific mutations responsible for blood pressure control. The thiazide sensitive Na-Cl cotransporter (NCC) has been implicated in the control of blood pressure, however a direct link between the kidney NCC and blood pressure regulation is lacking. Here, we report a case of chimerism in which a kidney from a patient with Gitelman syndrome was transplanted into a non-Gitelman hypertensive recipient. After transplantation, postural hypotension resulted, necessitating discontinuation of all antihypertensive medications used for treatment of calcineurin-induced hypertension. This is the first reported case of acquired Gitelman syndrome after transplantation. Transplantation of a Gitelman "kidney" into a hypertensive recipient provides additional support for the role of the kidney NCC in blood pressure regulation. Furthermore, this case suggests the potential use of thiazide diuretics in the treatment of calcineurin-induced hypertension. PMID- 16479188 TI - Palliative ethanol injections of unresectable advanced esophageal carcinoma combined with chemoradiation. AB - Combined chemoradiation therapy has proven to be an effective treatment for unresectable esophageal cancer. Nonsurgical endoscopic palliation of local disease has become feasible with neodymium:yttrium-aluminum-garnet laser, BICAP tumor probe, and metallic stents. Alternatively, endoscopic injections of ethanol are safe, inexpensive, and useful for palliation of malignant dysphagia. Two patients with unresectable squamous cell carcinoma of the esophagus were treated with 1 mL of absolute (95 g/L) alcohol injections once a week for 4 weeks, followed by chemoradiation therapy consisting of concomitant 5-fluorouracil 300 mg/m/d and radiation therapy (total of 60 Gy over 6 weeks). One patient had a complete response but died of alcoholism 25 months after diagnosis without evidence of tumor recurrence. The other patient had a partial response but died 16 months after diagnosis from disease progression. We conclude that tumor ablation by ethanol injection for palliation combined with chemoradiation may be a low-cost alternative for advanced unresectable esophageal cancer. PMID- 16479189 TI - Is chronic fatigue syndrome associated with platelet activation? AB - Chronic fatigue syndrome (CFS) is a debilitating condition that has no known aetiology or pathophysiology. Recent investigations by other workers have suggested that individuals with CFS may have a hypercoagulable state. This study investigated various aspects of platelet activation and function in 17 patients with CFS and in 16 age-matched and sex-matched healthy controls. Platelet aggregation, platelet volume and coagulation tests were performed. Platelet aggregation was investigated by means of the photometric changes using citrated platelet-rich plasma, whole blood aggregation was calculated as the percentage fall in single platelet counts and the coagulation tests were performed on an automatic microcentrifugal analyser.A trend was observed for the patients to have lower aggregation results and a reduced mean platelet volume. However, this only reached statistical significance for one result; the rate of the aggregation slope by 1.0 microg/ml collagen [CFS patients, 18 (9-28) versus controls, 32.5 (19-36); Mann-Whitney U test, P = 0.029]. No significant differences were found for any of the measurements of coagulation. These results are in contrast to previously reported findings. However, due to the heterogeneous nature of the disease, and the resulting lifestyles of the patients, caution should be taken when comparing one group of patients with another. Nevertheless, we certainly found no evidence of increased platelet activation or of a hypercoagulable state in patients with CFS and, on the basis of these results, anti-platelet or anti coagulant therapy is not warranted. PMID- 16479190 TI - The COX-2 G/C -765 polymorphism may modulate the occurrence of cerebrovascular ischemia. AB - In the atherosclerotic plaque, cyclooxygenase-2 (COX-2) catalyzes prostaglandin E formation, which acts as a pro-atherogenic factor. A polymorphism, G/C -765, within the COX-2 promoter region modulates gene expression and the risk of cerebrovascular disease. We have evaluated the relation between COX-2 G/C -765 genotypes and the occurrence of cerebrovascular ischemia. We evaluated the COX-2 G/C -765 polymorphism in 110 consecutive patients with a documented history of acute ischemic cerebrovascular disease, in 110 age-matched and sex-matched subjects without such history, and in a general population (n = 324) from the same ethnical background. The frequency of the COX-2 -765C allele in patients [0.21; 95% confidence interval (CI), 0.16-0.26] was similar to those found in controls (0.28; 95% CI, 0.22-0.34) and in the general population (0.26; 95% CI, 0.23-0.29). Carriers of the CC genotype differed between patients (0.02; 95% CI, 0.00-0.05) and controls [0.10 (95% CI, 0.04-0.16), P = 0.019; odds ratio, 0.17 (95% CI, 0.04-0.79)] or the general population [0.08 (95% CI, 0.05-0.11), P = 0.023; odds ratio, 0.22 (95% CI, 0.05-0.95)]. In a multiple logistic regression analysis adjusted for confounding variables, smoking status (P < 0.001), atrial fibrillation (P = 0.004) and COX-2 G/C-765 polymorphism (P = 0.016) independently contributed to cerebrovascular ischemia, with CC carriers exhibiting a lower risk (odds ratio, 0.07; 95% CI, 0.01-0.61). Our data show an association between the COX-2 G/C-765 gene polymorphism and cerebrovascular ischemia, suggesting that the COX-2 gene is a susceptibility locus for the risk of cerebrovascular ischemic disease. PMID- 16479192 TI - Direct action on the molecule is one of several mechanisms by which ultrasound enhances the fibrinolytic effects of reteplase. AB - Enhanced fibrinolytic reperfusion therapy may improve the outcome in embolic stroke, where ultrasound exposure has been shown to be one option. We recently verified that the fibrinolytic properties of streptokinase were modulated following ultrasound exposure of the molecule. We have now explored this possibility following ultrasound exposure of the reteplase molecule. The effects on clot lysis of reteplase and ultrasound both separately and in combination were studied by evaluating cumulated release of haemoglobin from whole blood clots following 1 h of exposure. Specifically, we investigated how clot lysis was modulated following pulsed 1 MHz ultrasound pre-exposure of the reteplase solution at intensities ranging between 0.125 and 4 W/cm2 spatial-average temporal-average intensity (SATA) and the effects of reteplase following 1 h of pre-exposure of clots to ultrasound at high intensity (4 W/cm2SATA). Significant enhancement of clot lysis during concomitant reteplase and pulsed ultrasound exposure were observed in two intensity ranges: 0.125-0.25 and 2-4 W/cm2SATA. Pre exposing reteplase solution to ultrasound significantly increased clot lysis only in the lower intensity range. At high ranges, pre-exposure of clots to ultrasound was followed by an increased fibrinolytic action of reteplase. Pre-exposing reteplase solution to low-intensity ultrasound induced changes in the reteplase molecule that enhanced its fibrinolytic effects. Although this effect disappeared at moderately higher ultrasound intensity, the pre-exposure of clots to ultrasound of higher intensity induced increased fibrinolytic effects of reteplase solution. PMID- 16479191 TI - The effects of unfractionated heparin, low molecular weight heparin and danaparoid on the thromboelastogram (TEG): an in-vitro comparison of standard and heparinase-modified TEGs with conventional coagulation assays. AB - To investigate the effects of unfractionated heparin (UFH), low molecular weight heparin (LMWH) and danaparoid (DPD) added to whole blood in vitro on standard and heparinase-modified thromboelastogram (TEG) parameters compared with conventional assays of coagulation. The effects of UFH, LMWH and DPD on standard TEG parameters were compared with the prothrombin time, activated partial thromboplastin time, thrombin time and anti-activated factor X (anti-FXa) activity, at concentrations of these anticoagulants ranging from 0.025 to 1 U/ml. In the second part of the study, the effects of very low concentrations (0.005 0.05 U/ml) of UFH, LMWH and DPD on the difference between standard and heparinase modified TEG parameters were compared with the prothrombin time, activated partial thromboplastin time, thrombin time and anti-FXa activity. Standard TEG parameters were outside the reference range at lower concentrations of UFH, LMWH and DPD than most conventional coagulation assays were able to detect. Only anti FXa activity was more sensitive to the presence of these anticoagulants than the standard TEG alone. The lowest concentration of UFH, LMWH and DPD used in this study (0.005 U/ml) caused significant differences between the standard and heparinase-modified alpha-angles of the TEG. In addition, the difference between standard and heparinase-modified TEG parameters distinguished between low concentrations (0.005-0.05 U/ml) of UFH with greater sensitivity than anti-FXa activity, but were less sensitive to LMWH and DPD. The standard TEG is more sensitive to UFH, LMWH and DPD than most conventional coagulation tests, with the exception of anti-FXa activity. Calculation of the difference between standard and heparinase-modified TEG parameters greatly increases the sensitivity of the assay for the effects of these anticoagulants, and is more sensitive to very low quantities of UFH than anti-FXa activity. PMID- 16479193 TI - High glucose contributes to aspirin insensitivity in streptozotocin-diabetic rats: a multiparametric aggregation study. AB - The effect of chronic hyperglycaemia on blood platelet response to acetylsalicylic acid was studied in rats with experimental diabetes. Platelet aggregation was determined in non-diabetic and streptozotocin-diabetic rats treated orally with 4 or 40 mg aspirin (ASA)/kg per day (for 8 weeks from the eighth day of diabetes) using whole blood impedance aggregometry with arachidonic acid or ADP as platelet agonists. The dose-dependent effect of ASA 'therapy' on ADP-agonized platelets was significant only in non-diabetic animals, while in diabetic rats both doses were ineffective in reducing ADP-stimulated platelet aggregation. ASA-mediated increased acetylation of platelet proteins favoured reduced platelet aggregation and slower platelet disaggregation (Pr < 0.025 or less). Interestingly, however, the occupation of platelet protein-free amino groups was significantly higher in control rats compared with diabetic rats (P < 0.001), pointing out that proteins of platelets in non-diabetic animals were more vulnerable for the ASA-induced acetylation. We conclude that chronic hyperglycaemia interferes with preventive effects of ASA on platelet reactivity. Our data validate the suggestion that the relationship between aspirin ineffectiveness and poor metabolic control, first revealed in humans, concerns also other animals' platelets and holds regardless of the model or type of diabetes. PMID- 16479194 TI - Effect of four missense mutations in the factor XIII A-subunit gene on protein stability: studies with recombinant proteins. AB - Four missense mutations in the factor XIII A-subunit gene, Arg260Leu, Ala318Val, Thr398Asn and Gly210Arg, were previously reported by us in patients with severe factor XIII deficiency. The objective of our study was to discern the effect of all four mutations on the stability and intracellular localization of the factor XIII A-subunit by their expression in COS cells. In-vitro mutagenesis, transient expression of the mutants in COS cells and subsequent pulse-chase analyses were carried out. Intracellular localization of wild-type and mutant proteins was analyzed by immunohistochemistry using a monoclonal antibody against factor XIII A-subunit. Pulse-chase analyses of metabolically labeled proteins demonstrated rapid intracellular degradation of each mutant protein as compared with wild type. Immunocytochemical and immunofluorescence analyses disclosed that wild-type and all four mutant factor XIII A-subunit proteins were diffusely distributed within the cytoplasm but not in the endoplasmic reticulum of the COS-7 cells. The Arg260Leu, Ala318Val, Thr398Asn and Gly210Arg mutations in FXIII A-subunit cause rapid intracellular degradation of the corresponding mutated protein. PMID- 16479195 TI - The effect of different anticoagulants on thrombin generation. AB - Decrease in thrombin generation is the key effect in anticoagulation. The aim of the present study was to investigate the effect of anticoagulants on thrombin generation and the relation to platelet count. Plasma samples from 10 healthy volunteers (mean age 43.0 +/- 9 years) were incubated at preset platelet counts with different doses of the anticoagulants lepirudin, fondaparinux and low molecular weight heparins. Thrombin generation was measured in a tissue factor mediated assay using a fluorometer and a slow-reacting fluorogenic substrate. The endogenous thrombin potential, the lag phase, the maximum reaction velocity (Vmax) and the concentration of a given anticoagulant required for 50% inhibition of thrombin generation (IC50) are presented. All three anticoagulants decreased endogenous thrombin potential and prolonged the lag phase in a dose-dependent manner. Fondaparinux and low molecular weight heparins, but not hirudin, decreased Vmax in a concentration-dependent manner. With increasing platelet count, the IC50 increased but the extent of this increase was not uniform for the three anticoagulants and the three variables investigated. The influence of anticoagulants on thrombin generation is variable, depending on their basic mechanism of action. In defining and comparing their effects, the endogenous thrombin potential, the lag phase and the maximum reaction velocity should be considered together. Platelets have a considerable influence on the magnitude of thrombin generation. PMID- 16479196 TI - Changes in hypercoagulability by asparaginase: a randomized study between two asparaginases. AB - Alterations in hemostasis have frequently been observed in children with acute lymphoblastic leukemia. Thrombotic events are well documented in patients receiving L-asparaginase as a single agent or in combination with other chemotherapeutic drugs. The present prospective, randomized study evaluated the effect of two different L-asparaginase preparations, native Escherichia coli L asparaginase (Crasnitin; Bayer AG, Leverkusen, Germany; n = 10) and L asparaginase derived from Erwinia chrysanthemi (Erwinase; Porton Pruducts, London, UK; n = 10) on the changes in parameters concerning hypercoagulability. Patients were randomized to receive a total of eight doses of 10,000 IU/m2 L asparaginase intravenously with intervals of 3 days during induction therapy. Before starting L-asparaginase treatment all patients had already demonstrated an increased thrombin generation shown by the elevated levels of prothrombin fragment 1+2 and thrombin antithrombin III, presumably due to therapy with prednisone, daunorubicin and vincristine. A significant decrease in alpha2 antiplasmin and plasminogen levels was measured in the E. coli L-asparaginase but not in Erwinase-treated patients. Increased thrombin generation combined with a decrease in alpha2-antiplasmin and plasminogen levels may lead to a state of increased risk for thrombosis due to a delay in fibrin elimination in E. coli L asparaginase-treated patients only. PMID- 16479198 TI - Successful use of recombinant factor VIII devoid of von Willebrand factor during multiple teeth extractions in a patient with type 3 von Willebrand disease. AB - We report a 55-year-old patient with type 3 von Willebrand disease who underwent multiple tooth extractions with successful hemostatic management using recombinant factor VIII. The patient was previously misdiagnosed and treated incorrectly then at 53 years old, he was diagnosed with type 3 von Willebrand disease. As he had avoided dental treatments for two decades due to severe bleeding after dental extraction, multiple severe caries and marginal periodontitis were revealed. The patient refused the use of blood products in hemostatic management because he was afraid of blood-borne diseases and development of anti-von Willebrand factor alloantibodies. After close consultation, we therefore decided to use recombinant factor VIII. Four teeth extraction procedures were executed twice. Before extraction, bolus recombinant factor VIII (50 IU/kg) was administered intravenously followed by continuous infusion (5-10 IU/kg per h) for approximately 48 h. The factor VIII:C level increased from about 1 to 20-32% 30 min after bolus infusion. During continuous infusion (10 IU/kg/h), factor VIII:C was maintained at more than 10%. Little bleeding occurred during and after the multiple teeth extractions and during suture removal. On frequent examinations during a 1-year follow-up, neither von Willebrand factor nor factor VIII inhibitors were detected. PMID- 16479197 TI - Fondaparinux as anticoagulant in a pregnant woman with heparin allergy. AB - We report a patient who had a history of deep vein thrombosis in a previous pregnancy. She was treated with heparins without any reactions in the index pregnancy. Subsequently, when the patient became pregnant again, she developed an acute cutaneous reaction to the low molecular heparin enoxaparin 3 weeks after initiation of therapy. She developed a similar reaction to delteparin as well. She was therefore treated with warfarin until 36 weeks of gestation. Then she was treated with fondaparinux (Arixtra, Sanofi-Synthelabo, Paris, France) 2.5 mg daily for the remainder of the pregnancy. Delivery was at term by induction of labour. Fondaparinux was stopped on the day of the induction of labour. It was re started 6 h post-delivery and the patient was anticoagulated with warfarin in the post-partum period. There were no bleeding tendencies or recurrences of thrombosis during fondaparinux therapy. Both mother and baby were well after delivery. PMID- 16479199 TI - A 2-year retrospective analysis of laboratory testing for activated protein C resistance with a factor V-corrected activated partial thromboplastin time-based method. AB - The factor V-corrected activated protein C resistance assay is the test of choice to screen for the factor V Leiden mutation. During the past 2 years, local test results with the frequently used Coatest APCR kit were evaluated and compared with the results of DNA analysis, the 'gold standard'. Samples of 278 patients were analysed by both techniques. We were unable to confirm that factor V Leiden carriers can clearly be delineated from normal individuals with the Coatest APCR test. A ratio of 2.0 as the cut-off provides 99.0% sensitivity and 95.4% specificity.To evaluate the lupus anticoagulant interference, we retrospectively analysed 16 lupus anticoagulant-positive patients. In this study, two (12.5%) showed a false-positive activated protein C resistance result. Six out of 16 (37.5%) lupus anticoagulant-positive patients were also carriers of the factor V Leiden mutation. Four out of eight (50%) false-positive activated protein C resistance results presented with an abnormal baseline clotting time. In order to prevent reporting false-positive results, a maximum baseline clotting time (65.8 s) was calculated. A new scheme for interpreting activated protein C resistance ratios was proposed. PMID- 16479200 TI - In-vitro model for the ultrastructural study of the formation of thrombi in human platelets. AB - Platelets are cell fragments with dynamic properties involved in clot formation after tissue damage. Platelet activation causes a change in shape, secretion of intracellular granules and aggregation with each other through the cytoskeleton components and biochemical changes. Platelet adhesion, considered as the major event in haemostasis, has been studied in several in-vitro and in-vivo models to evaluate the feasible thrombogenicity of some materials, the dynamics of specific receptors, as well as the effect of different buffers and inhibitors in this process. In spite of the numerous reports about platelet activation, to date there is no information available about the fine structure of the platelet platelet and platelet-substrate interactions. In the present report we describe an in-vitro system that allows the visualization of these interactions: platelets are adhered to an inert substrate, and interactions with suspended platelets as a process to initiate the formation of thrombi was followed by ultramicrotomy and transmission electron microscopy. PMID- 16479202 TI - Laparoscopic nephron sparing surgery: ischemic renal damage. PMID- 16479203 TI - Laparoscopic adrenalectomy for adrenal carcinoma and metastases. AB - PURPOSE OF REVIEW: This review article discusses topics concerned with laparoscopic adrenalectomy for adrenal pathologies. RECENT FINDINGS: Over the past decade, laparoscopic adrenalectomy has become the operation of choice for the resection of adrenal tumors, and provides dramatically reduced morbidity associated with the operation. Although current laparoscopic adrenalectomy for metastatic or primary adrenal malignancy is a feasible procedure, great care is required. Laparoscopic adrenalectomy for these adrenal pathologies should be converted to open adrenalectomy or hand-assisted laparoscopic adrenalectomy for difficult dissection, invasion, adhesions or surgeon inexperience. In general, laparoscopic adrenalectomy can be performed with acceptable outcomes in carefully selected patients with small, organ-confined, solitary adrenal metastasis or primary adrenal carcinoma. SUMMARY: As surgical technique and complete resection are crucial to the success of the procedure, appropriate and careful judgment on the part of the surgeon is the most important factor in the care of a patient with such adrenal malignancies. PMID- 16479204 TI - Female incontinence: a review of biomaterials and minimally invasive techniques. AB - PURPOSE OF REVIEW: This article discusses the various grafts or biomaterials, minimally invasive techniques, and recent advances for the treatment of female stress urinary incontinence and pelvic organ prolapse. RECENT FINDINGS: The studies reviewed in this paper compared certain biologic grafts to synthetic grafts in clinical trials and histopathological studies. Data from long-term outcome studies for tension-free vaginal tape are evaluated. As tension-free vaginal tape is the foremost technique for stress urinary incontinence correction, many of the newer modalities such as transobturator tape and laparoscopy are compared with it. Immediate and long-term complications from mesh use in stress urinary incontinence and pelvic organ prolapse repair are examined. Correction of prolapse may eventually entail the use of specially designed 'kits' that allow total pelvic floor reconstruction with a single piece of mesh. SUMMARY: Although biological grafts are initially efficacious, the trend is to use synthetic grafts in repair of stress urinary incontinence and pelvic organ prolapse. Midurethral slings continue to be the front-line therapeutic modality for stress urinary incontinence. After analysis of long-term data, other surgical techniques may gain popularity. With increasing use of synthetic grafts, however, long-term complications such as de-novo urgency, erosion, and dyspareunia need to be assessed. PMID- 16479205 TI - Incontinence after radical prostatectomy: surgical treatment options. AB - PURPOSE OF REVIEW: Stress urinary incontinence is an uncommon finding in healthy men. Following radical prostatectomy, however, stress urinary incontinence is the most important complication influencing patient morbidity following surgery and influencing satisfaction, regardless of the surgical technique applied. Conservative treatment options centre at early restoration of bladder and pelvic floor function. When significant and bothersome stress urinary incontinence persists, even after six to 12 months active treatment follow-up, surgical restoration of continence seems advisable. RECENT FINDINGS: Two themes emerge in reviewing the recent publications in this field. Implantation of the hydraulic artificial urinary sphincter is still the gold standard, particularly in severe cases. Alternatively, new techniques and other artificial materials are gaining favour, aiming at a large cohort of patients with less severe incontinence, which have been not treated or overtreated so far. Advances in both areas are covered within this review article in detail. SUMMARY: For treatment of incontinence following prostatectomy, a large variety of surgical techniques are readily available and have proven to be helpful tools in making patients' uncomfortable lives much easier. In addition, the wide armamentarium of artificial materials and techniques may help to choose the proper surgical technique for every patients' needs. PMID- 16479206 TI - Training in laparoscopic urology. AB - PURPOSE OF REVIEW: Training in laparoscopy has become an important issue in the current surgical scenario. In this overview we aim to update the current knowledge in the field of laparoscopic urological training and to highlight the potential dangers of using simulation for accreditation and selection purposes at this stage. RECENT FINDINGS: Physical simulators are widely available and seem to be equally efficient as virtual reality simulators. Transfer of training has been proven to be beneficial in randomized controlled trials for virtual reality and cholecystectomy. A model for the vesico-urethral suture has been described and integrated in a skills laboratory program. The program has construct validity and can discriminate at least between beginners and advanced laparoscopists. Efforts have still to be made in defining appropriate tools to assess competence and evidence for reliability, and validity must be obtained before including simulators in accreditation programs. SUMMARY: In spite of the abundant literature there is still little evidence about the learning mechanism involved in acquiring laparoscopic skills. Physical and virtual reality simulators have been proven to be efficient in improving dexterity and some evidence exists of a positive transfer from virtual reality to the operating room in cholecystectomy. Very few models, however, have been described for reconstructive urology, and effective transfer to the operating room has not yet been proven, although validation work is in progress in the field of urology. PMID- 16479207 TI - The management of ureteric colic. AB - PURPOSE OF REVIEW: The management of ureteric colic has changed significantly over the past three decades,. Factors such as stone size, location, composition as well as patient and surgeon preference influence the management decision. RECENT FINDINGS: Recent work has looked at the expansion of the role of medical therapy beyond symptomatic control to attempt to target some of the reversible factors in stone retention and thereby improve the likelihood of spontaneous stone expulsion. Changes in shock wave lithotripsy and ureteroscopy offer patients highly effective, minimally invasive options for stone clearance that can often be performed on an outpatient basis. SUMMARY: This paper reviews the options available to the urologist in the treatment of ureteric colic as well as the advantages and disadvantages of each therapy. PMID- 16479208 TI - Minimizing symptoms in patients with ureteric stents. AB - PURPOSE OF REVIEW: The current approaches for minimizing symptoms in patients with ureteric stents were reviewed utilizing a literature search on Pubmed using the keywords stent, symptom, and ureter. RECENT FINDINGS: Ureteral stents are widely used in urological procedures for maintaining upper urinary tract drainage to relieve obstruction, pain, or infection. Indwelling stents, however, are associated with significant morbidity such as infection, encrustation, hematuria, and bothersome symptoms. Minimizing these issues has become paramount in the design of new ureteral stents. This article will review current and novel ways to minimize stent-related morbidity. SUMMARY: Currently, there is no ideal stent that relieves obstruction, is resistant to infection and encrustation, and is comfortable for patients. Advances in biomaterials and design will result in a more biocompatible stent that also has patient comfort in mind. PMID- 16479209 TI - Effect of modification of shock-wave delivery on stone fragmentation. AB - PURPOSE OF REVIEW: Shock-wave lithotripsy has been the mainstay of urinary-stone treatment over the past 20 years, with three generations of lithotripters now in the market place. Little improvement, however, has been made in the overall efficiency, since the original Dornier HM3 lithotripter. Over the past 5 years much progress has been made in the basic research of shock-wave lithotripsy, with better understanding of the mechanisms involved in stone fragmentation. This progress has led to new modifications in the way shock-wave pulse is generated and delivered. RECENT FINDINGS: Clinical studies, reflecting improved understanding of basic mechanisms of stone comminution, are being published. Two recent prospective clinical trials have shown the higher efficiency of slow-rate compared with fast-rate shock-wave lithotripsy. A very practical solution requiring no hardware upgrade albeit at longer procedure times. Other promising developments include the use of twin-head technology, with either simultaneous or sequential shock waves. In addition, chemolytic pretreatment and dose-escalation techniques have shown early encouraging results. This review provides an update of the latest shock-wave technology and delivery strategies. SUMMARY: Long-term studies, to document anticipated improved safety with slow shock-wave rate, are needed. Future in-vivo and clinical studies of twin-head technology and dose escalation strategy of shock-wave lithotripsy may initiate new lithotripter designs that will lead to improved stone-free rates, while simultaneously reducing associated renal trauma. PMID- 16479210 TI - Prediction of shockwave failure in patients with urinary tract stones. AB - PURPOSE OF REVIEW: To evaluate recent developments in predicting the failure of shockwave lithotripsy when treating patients with urinary tract stones. RECENT FINDINGS: Although the features of patients with stones, as well as of the stones themselves, associated with refractoriness to shockwave lithotripsy are fairly well defined, refining the preoperative detection of these traits and optimizing the efficacy of shockwave lithotripsy are still under investigation. Several studies have recently focused on improving the radiological appraisal of stone size and composition through the use of axial computed tomography and reconstruction software. Other investigators have researched techniques to increase the efficacy of the technology underlying shockwave lithotripsy, such as varying the shockwave delivery rate and method. SUMMARY: Investigators have demonstrated different factors and predictors that affect shockwave lithotripsy for stone disease. Continued research will better define patient selection and the role of shockwave lithotripsy in the treatment of urolithiasis. PMID- 16479211 TI - Urolithiasis in patients with spinal cord injuries: risk factors, management, and outcomes. AB - PURPOSE OF REVIEW: Despite major advances in the urological care of spinal cord injury patients, the incidence of urolithiasis has not changed. Although the incidence of kidneys lost to staghorn calculus disease has decreased considerably, stone disease is still a significant cause of morbidity and concern. The purpose of this review is to summarize the risk factors for the development of stone diseases in the spinal cord injury patient population and present the outcomes of current endourologic treatment modalities. RECENT FINDINGS: Recurrent urinary tract infections, indwelling catheters, vesicoureteral reflux, and immobilization hypercalcuria are a few of the major risk factors for the development of urolithiasis among spinal cord injury patients. Retrograde endourologic techniques are often not possible to address stone disease due to lower extremity contractures, spinal curvature, and pelvic tilt. Extracorporeal shockwave lithotripsy success rates vary from 50 to 90%, but clearance rates are often delayed. Success rates for percutaneous nephrolithotomy match those quoted in the general population ( > 90%), but at the expense of a higher rate of complications (6-20%). Meticulous planning with regard to appropriate prophylactic antibiotics and body position will maximize efficacious outcomes. SUMMARY: Early identification and treatment of urolithiasis in spinal cord injury patients will aid in preserving renal function and minimizing associated complications. Despite variation in common urological practices between spinal cord injury units and the lack of clear cut guidelines for follow up, the increased incidence of risks associated with urolithiasis lends support for routine genitourinary imaging in order to identify and treat those individuals at highest risk. PMID- 16479212 TI - Management of stones in patients with anomalously sited kidneys. AB - PURPOSE OF REVIEW: Congenital abnormities in urology are very common. Horseshoe, malrotated and ectopic kidneys, as well as duplex systems, are the most frequent in this respect. The combination of both abnormalities and stones is of clinical importance. The question is asked if standard procedures for stones apply also to stones in abnormal kidneys. RECENT FINDINGS: In general, open surgery, extracorporeal shock-wave lithotripsy, percutaneous procedures, endoscopic procedures and laparoscopy are possible procedures in both normal and abnormal kidneys. The importance of ureteric pelvic junction obstruction has to be taken into account and a metabolic work-up remains important. SUMMARY: The trend for treatment of stones in abnormal kidneys goes towards endoscopical and laparoscopical procedures, whereas a combination of both seems to be appropriate in many cases. PMID- 16479214 TI - Bibliography. Current world literature. Minimally invasive surgery in urology. PMID- 16479213 TI - Is there still a role for open surgery in the management of renal stones? AB - PURPOSE OF REVIEW: In the last few decades, with the improvement in endourological surgery and the invention and evolution of extracorporeal shock wave lithotripsy, the indications for open surgery in stone disease have become rare, although open surgery still has a role in selected cases. In this review we discuss the current indications of open surgery for the elimination of urinary calculi. RECENT FINDINGS: A MEDLINE and MeSH search was performed to evaluate currently available guidelines on open stone surgery and identify the evidence based medicine that support the role of open surgery in treating urinary lithiasis. The latest papers published on open stone surgery are reviewed and conclusions are drawn, based on their results. SUMMARY: Open stone surgery should be avoided in most cases, but should be considered for those patients in whom a reasonable number of less invasive procedures would not be useful. The most common indications for open stone surgery include complex stone burden, failure of extracorporeal shock-wave lithotripsy or endourological treatment and anatomical abnormalities (such as ureteropelvic junction obstruction and infundibular stenosis with or without renal caliceal diverticulum). The level of evidence for the currently available guidelines is not adequate, mainly because of lack of properly designed, large prospective randomized trials that compare different treatment options. PMID- 16479215 TI - Biological basis of fracture healing. PMID- 16479216 TI - Nonunion treatment: iliac crest bone graft techniques. AB - In the management of nonunions, detailed surgical treatment plans will vary depending on the underlying etiology of the specific case. Iliac crest autogenous bone grafting, although associated with donor site complications, continues to be a necessary part of the treatment of many nonunions. This article summarizes the classification of nonunions and the use of iliac crest autogenous bone grafting, the standard against which any new technique must be measured. PMID- 16479217 TI - Overview of biologics. AB - A variety of orthobiologic materials are available or under development to enhance the repair of bone fractures. These materials vary widely in their composition, mechanism of action, and clinical results. This article provides a brief review of the currently available bone grafting materials, which include autografts, allografts, conductive substrates, demineralized bone matrix, platelet gels, recombinant bone morphogenic proteins, and bone marrow aspirate injections. PMID- 16479218 TI - Bone morphogenic protein science and studies. AB - The use of growth factors, such as bone morphogenic proteins (BMPs), is showing great promise in orthopaedic surgery. The BMPs, which are osteoinductive, have been used successfully in spine fusion and fracture healing. This article describes the development and production of the recombinant BMPs and summarizes their use in preclinical trials. PMID- 16479219 TI - Recombinant human bone morphogenic protein-2 in fracture care. AB - Recombinant human bone morphogenic protein-2 (rhBMP-2) is an osteoinductive protein that is now available for the acute treatment of open tibial fracture. Data from preclinical and clinical trials support its role in fracture care. This article summarizes data from two randomized controlled clinical trials investigating the safety and efficacy of rhBMP-2 in patients with open tibial shaft fractures and tibial fractures with bone defects. Also included is a summary of an economic analysis that examined the cost impact of rhBMP-2 from a hospital and payor perspective. PMID- 16479220 TI - Recombinant human bone morphogenic protein-7: applications for clinical trauma. AB - Several clinical trials have examined the safety and efficacy of recombinant human bone morphogenic protein-7 (rhBMP-7) for osseous repair in clinical trauma. This article summarizes the results obtained from three recent clinical trials, which suggest that rhBMP-7 is useful in the treatment of tibial nonunion, long bone nonunion, and open tibial shaft fractures. PMID- 16479221 TI - The science of fracture healing. AB - Fracture healing is a unique repair process in which the events of endochondral and intramembranous bone formation follow a specific time sequence. There are many ways in which these events can be disrupted, leading to unsuccessful fracture healing. This article summarizes the series of cellular and molecular events that take place in the process of endochondral fracture healing, based on observations obtained from an animal model of a femoral fracture. PMID- 16479222 TI - Initial fracture management and results. AB - In the management of fractures, the appropriate treatment must take into consideration the nature of the injury, the surgical insult and technique, and a variety of patient factors. The surgeon must also anticipate potential complications and plan the initial treatment accordingly. This article discusses the impact of surgical decisions on the process of fracture healing. PMID- 16479223 TI - Responding to the changing needs of public health assessment in the information age: the evolution of a program. PMID- 16479224 TI - Community health assessment: driving the need for current, easily accessible population health data. AB - Community health assessment (CHA) is a core public health function. Community health assessment is a term that describes both a process and its tangible products, such as a community health profile or other types of reports on the status of the community's health and existing influences on its health, including the extent and nature of health-related resources. Assessment has enabled a better understanding of the community's health, and this has impacted public health decisions. The authors use the framework for community health improvement proposed in 1997 by the Institutes of Medicine to show how CHA is part of the community health improvement process. Community health assessment drives the need for current, easily accessible population health data. The importance of Web based data query systems, the focus of the other articles in this special issue, can be best understood within the broader context of CHA. Selected case examples of how Web-based data query systems have impacted the CHA process in three states are highlighted. PMID- 16479225 TI - Characteristics, desired functionalities, and datasets of state Web-based Data Query Systems. AB - Based on a review of state public health department Web sites, this article describes Web-based Data Query Systems (WDQSs) currently used in 27 US states. In addition, functionalities of selected well-established state WDQSs are defined as major functionalities (11), subfunctionalities (21), and specific functionalities (141), on the basis of a consensus process with participants from states and national organizations in the United States and Canada. Also, on the basis of the consensus process, specific functionalities are categorized as core, enhanced, and expert. Key terms relevant to WDQSs are defined. The information contained in this article should prove useful to public health agencies developing, revising, or evaluating the WDQS. PMID- 16479226 TI - Innovations in user-defined analysis: dynamic grouping and customized user datasets in VistaPHw. AB - Flexible, ready access to community health assessment data is a feature of innovative Web-based data query systems. An example is VistaPHw, which provides access to Washington state data and statistics used in community health assessment. Because of its flexible analysis options, VistaPHw customizes local, population-based results to be relevant to public health decision-making. The advantages of two innovations, dynamic grouping and the Custom Data Module, are described. Dynamic grouping permits the creation of user-defined aggregations of geographic areas, age groups, race categories, and years. Standard VistaPHw measures such as rates, confidence intervals, and other statistics may then be calculated for the new groups. Dynamic grouping has provided data for major, successful grant proposals, building partnerships with local governments and organizations, and informing program planning for community organizations. The Custom Data Module allows users to prepare virtually any dataset so it may be analyzed in VistaPHw. Uses for this module may include datasets too sensitive to be placed on a Web server or datasets that are not standardized across the state. Limitations and other system needs are also discussed. PMID- 16479227 TI - Implementation of an Internet-based geographic information system: the Florida experience. AB - Increasingly, Web-based data dissemination systems are being used to provide public health information. This article describes Florida's experience in augmenting a Web-based data query system, Florida CHARTS, with a geographic information system component. As with any systems development, users' needs guide the product. With a focus on community health assessment and improvement planning, several methods of data display have been established. A variety of issues are explored including the impact of small numbers of events on data display and data confidentiality, the selection of geographic units of analysis, and techniques for geographically coding data. PMID- 16479228 TI - Utah's IBIS-PH: an innovative user interface solution for Web-based data query systems. AB - The ability to make good Web-based data query system (WDQS) project management decisions requires an understanding of the trade-offs inherent among various technology options. This article presents the current options available for the user interface and data display and compares the advantages and disadvantages of each for use in a WDQS. Relevant options are also discussed for back-end technologies such as Web and application servers and data storage mechanisms. We explain our decisions in developing the Indicator-based Information System for Public Health (IBIS-PH) query system to increase the probability of success and minimize risk. Finally, we compare the resulting IBIS-PH application characteristics with our original design requirements: broad public access; rich, interactive, and easy-to-use interface; portability; accessibility; maintainability of software and interface; supportability; low cost; and security. PMID- 16479229 TI - Perspectives on the Massachusetts Community Health Information Profile (MassCHIP): developing an online data query system to target a variety of user needs and capabilities. AB - The Massachusetts Community Health Information Profile (MassCHIP) has many distinctive features. These features evolved to maximize the usefulness of this query system for a broad group of users with varied needs, differing levels of knowledge about public health, and diverse experience using public health data. Three major features of MassCHIP help target our large user population. These features are as follows: (1) multiple avenues of entry to initiate queries ranging from an alphabetical list of simple topics to detailed International Classification of Disease codes; (2) the inclusion of data sets from other state agencies in addition to those of the Massachusetts Department of Public Health to reflect a broad view of public health; and (3) the capacity to retrieve data for multiple levels of geography, from the neighborhood through the state, including planning districts and hospitals. In this article, we discuss the history and design of MassCHIP, and focus on the features of MassCHIP that target a great variety of user needs and capabilities, and which are distinctive among Web-based data query systems. PMID- 16479230 TI - Prioritization MICA: a Web-based application to prioritize public health resources. AB - Although setting priorities is an important step in making public health policy, the benefit of using epidemiology to prioritize scarce public health resources has not been fully recognized. This situation is mostly due to the complexity of proposed models for setting priorities. We describe a public health priority setting model, Missouri Information for Community Assessment Priority Setting Model (Priority MICA), which uses epidemiologic measures available in most surveillance systems across the United States. Priority MICA uses data from birth and death certificates, hospital discharges, emergency departments, risk factors from the Behavioral Risk Factors Surveillance System, and eight epidemiologic measures to construct six priority criteria: size (the number of emergency department visits, hospitalizations, and deaths), severity (number of deaths of people younger than 65), urgency (trends in deaths and hospital morbidity), preventability (evidence-based score), community support (score of social support for preventive action), and racial-disparity (race comparison through death and morbidity rate ratio). Priority MICA is part of a Web-based interactive tool that makes available data from a wide variety of surveillance systems (http://www.dhss.mo.gov/MICA). The top 10 priority diseases determined by Priority MICA were compared to a more traditional method of ranking diseases by mortality rates. Using the additional criteria in Priority MICA identified four more priority diseases than were identified using just mortality while the ranking of the other six priority diseases differed between methods. PMID- 16479232 TI - Small numbers, disclosure risk, security, and reliability issues in Web-based data query systems. AB - This article describes the process for developing consensus guidelines and tools for releasing public health data via the Web and highlights approaches leading agencies have taken to balance disclosure risk with public dissemination of reliable health statistics. An agency's choice of statistical methods for improving the reliability of released data for Web-based query systems is based upon a number of factors, including query system design (dynamic analysis vs preaggregated data and tables), population size, cell size, data use, and how data will be supplied to users. The article also describes those efforts that are necessary to reduce the risk of disclosure of an individual's protected health information. PMID- 16479233 TI - Reflections on organizational issues in developing, implementing, and maintaining state Web-based data query systems. AB - Emerging technologies, such as Web-based data query systems (WDQSs), provide opportunities for state and local agencies to systematically organize and disseminate data to broad audiences and streamline the data distribution process. Despite the progress in WDQSs' implementation, led by agencies considered the "early adopters," there are still agencies left behind. This article explores the organizational issues and barriers to development of WDQSs in public health agencies and highlights factors facilitating the implementation of WDQSs. PMID- 16479231 TI - States' approaches to implementing a Web-based data query system. AB - BACKGROUND: Individual states have made different decisions regarding Web-based data query systems (WDQSs). The goal of this study was to help state, county, or city health departments deploy or update a WDQS by showing how different decisions are guided by unique situations within states at particular times. METHODS: Interviews were held with key informants from 10 states that had a WDQS to determine factors relating to how these states implemented their WDQSs. RESULTS: Three states designed and developed their own systems, three states implemented systems directly using a Web-query software package, two states adopted systems developed by other states, one state designed and developed its own system after adopting a system, and one state used a Web-query software package to implement a system after attempting to adopt a system. None of the states used a commercial off-the-shelf system to implement a WDQS. All the WDQSs were located in the health statistics office or in an umbrella division of the public health agency that included the health statistics office. The skill set of available personnel had a significant impact on a state's implementation decision. CONCLUSIONS: All key informants reported satisfaction with their decisions regarding implementing a WDQS. This provides support for states currently considering any of these options. PMID- 16479234 TI - Measuring the impact of a Web-based data query system: the logic model as a tool in the evaluation process. AB - A review of program evaluation perspectives, including the concept of a logic model and its applicability to the evaluation of Web-based data query systems (WDQSs), is presented. A logic model is used to flesh out evaluation components of a WDQS, including inputs, constraints, program activities, program outputs, and outcomes. For each component, a list of potential items is presented for inclusion in the model, along with examples of initial, intermediate, and ultimate outcomes for a WDQS. Program evaluation is a process that is important to conduct both early in WDQS development to promote clarity of its vision and objectives and throughout the course of the WDQS implementation and maintenance to ensure that its objectives are being met. Should the WDQS not be producing desirable results, the logic model provides a road map for understanding which activities are not meeting the WDQS's objectives so that the course of WDQS development, implementation, and maintenance may be altered to improve the probability of reaching desirable outcomes. PMID- 16479235 TI - Envisioning futures for state Web-based data query systems. PMID- 16479236 TI - Community coalition project directors: what makes them effective leaders? AB - OBJECTIVE: Effective leaders, both voluntary and paid, facilitate successful coalitions. The attributes that characterize effective project directors, however, are unclear. Our aim was to identify characteristics of effective project directors leading community coalitions. METHODS: The study examined 13 project directors who led eight community-based coalitions established to combat substance abuse. We inductively identified common characteristics and leadership effectiveness of the project directors by abstracting data from detailed ethnographic studies of these coalitions. We assessed the validity of leadership effectiveness by comparing data abstracted from ethnographic studies with two independent ratings. We then employed a cross-case comparison strategy for analyzing patterns among the common characteristics identified and leadership effectiveness. Six characteristics emerged among the project directors studied: status with community (insider vs outsider); shared leadership; bridge building skills; substance abuse expertise; vision; and management style. RESULTS AND CONCLUSIONS: Shared leadership, bridge building skills, and insider status were consistently related to leadership effectiveness. Less support was found for substance abuse expertise or vision. When hiring project directors, coalition leaders may consider assessing whether candidates are "insiders" within the community and demonstrate shared leadership and bridge building skills. PMID- 16479237 TI - Another year of influenza vaccine supply and distribution problems: why does this keep happening and what can be done about it? PMID- 16479239 TI - Fever of unknown origin: a systematic review of the literature for 1995-2004. AB - BACKGROUND: Fever of unknown origin (FUO) identifies a pattern of fever with temperature higher than 38.3 degrees C on several occasions over more than 3 weeks, in which the diagnosis remains uncertain after an initial diagnostic work up. The identification of the cause of FUO is a challenge in clinical practice despite recent advances in diagnostic techniques. There are more than 200 reported causes of FUO and they can be classified in four diagnostic categories: infections, neoplasms, non-infectious inflammatory diseases and miscellaneous. METHODS: We performed a systematic research of the literature on classical FUO to retrieve the review articles and case series published from 1995 to 2004, including articles from developing countries. The case series were reviewed to identify the tests commonly used both to qualify a fever as FUO and to determine the cause of the FUO, and to design an updated flow chart for the diagnosis of classical FUO. RESULTS AND CONCLUSIONS: No standardized diagnostic strategy could be determined. The diagnostic process should be guided by the potential diagnostic clues (PDCs) emerging from the history, physical examination and baseline tests. A standardized flow chart can be applied only in absence of PDCs or when the PDCs are contradictory.Nuclear medicine techniques are a valuable aid in the search for the origin of FUO due to bacterial infections or in the absence of PDCs. PMID- 16479240 TI - Fever of unknown origin, infection of subcutaneous devices, brain abscesses and endocarditis. AB - The term 'fever of unknown origin' includes a wide range of conditions that often remain undiagnosed. The possibility of an infection must be promptly diagnosed in order to begin appropriate therapy. Imaging with radiopharmaceuticals, computed tomography, magnetic resonance imaging and ultrasound are the most commonly applied techniques, usually performed in addition to blood tests, biopsies or tissue cultures when required. The lack of comparative studies investigating the accuracy of each radiopharmaceutical for the study of fever of unknown origin was the incentive to perform a meta-analysis of peer articles published between 1981 and 2004 (33 papers) describing the use of nuclear medicine imaging for this purpose. Furthermore, infection of subcutaneous devices, brain abscesses and endocarditis must be considered amongst the causes of fevers of unknown origin. Reviews of 23, 10 and 10 papers, respectively (from 1976 to 2005), were performed on these specific topics. The results may be a useful guide for the choice of the optimal radiopharmaceutical(s) and diagnostic strategy to be applied in each clinical condition and for different aims. PMID- 16479241 TI - Clinical applications of 188Re-labelled radiopharmaceuticals for radionuclide therapy. AB - 188Re is a radionuclide in which there is widespread interest for therapeutic purposes because of its favourable physical characteristics. Moreover, it can be eluted from an on-site installable 188W/188Re generator, which has a useful shelf life of several months. Most of the clinical experiences gained with 188Re concern the use of 188Re-1,1-hydroxyethylidenediphosphonate (188Re-HEDP) for bone pain palliation in patients suffering prostate cancer. The maximum tolerated activity was 3.3 GBq 188Re-HEDP and if the platelet count exceeded 200 x 10(9) l( 1), the administration of 4.4 GBq appeared safe. Evidence for repeated administrations of 188Re-HEDP rather than single injections was established. In general, pain palliation occurs in 60-92% of patients with only moderate transient toxicity, mainly related to changes in blood counts. Also in haematology, radioimmunotherapy by means of 188Re might play a role by selectively targeting the bone marrow in patients undergoing conditioning prior to haematopoetic stem cell transplantation. The feasibility of such an approach was proven using a Re-labelled monoclonal antibody directed toward the CD66 antigen. More recently, encouraging safety data on locoregional treatment of primary liver tumours using 188Re-labelled lipiodol were reported. The normal organs at greatest risk for toxicity are the normal liver and the lungs. About 50% of the patients reported mild and transient side effects, mainly consisting of low grade fever, right hypochondrial discomfort or aggravation of pre-existing liver impairment. Besides the applications in oncology 188Re-based therapies have also been pioneered for benign condition such as prevention of re-stenosis following angioplasty and for radiosynovectomy in cases of refractory arthritis. PMID- 16479242 TI - FDG PET positive lymph nodes are highly predictive of metastasis in breast cancer. AB - AIM: To determine whether or not fluorodeoxyglucose positron emission tomography (FDG PET) imaging when positive could obviate the necessity for sentinel lymph node biopsy and for complete axillary node dissection in patients with breast cancer. METHODS: A total of 80 female patients with a histological diagnosis of breast cancer and clinically negative axillary nodes underwent an FDG PET and sentinel lymph node biopsy (SLNB) or total axillary dissection for staging of axilla. Both SLNB and axillary dissection were performed in 72 patients, while eight patients had total axillary dissection without SLN biopsy. RESULTS: Of the 80 patients, 36 had lymph node metastasis on histopathology. SLNB was positive for metastasis in 35 (97%) of 36 patients (29 macrometastasis and seven micrometastasis). In the patient with false negative SLNB, the lymph node was completely replaced by the tumour. The FDG PET was true positive in 16 of 36 patients (sensitivity, 44%). There were two false positive studies with FDG PET, resulting in a specificity of 95%. The positive predictive value and accuracy of FDG PET for the detection of axillary lymph node metastasis were 89% and 72%, respectively. Univariate analysis revealed that higher grade of tumour, increased size and number of axillary lymph nodes were significantly associated with positive FDG PET results for axillary staging. CONCLUSION: FDG PET cannot replace histological staging using SLNB in patients with breast cancer. However, FDG PET has a high specificity and positive predictive value for staging of the axilla in these patients. The patients with higher grade of tumour, larger size and higher number of axillary lymph nodes may be considered for FDG PET scan for axillary staging. PMID- 16479243 TI - Three-dimensional positron emission tomography imaging with 124I and 86Y. AB - BACKGROUND: Impure positron emitters have physical characteristics that degrade image quality compared to conventional positron emitters like 18F. Two impure positron emitters with potentially interesting applications are 124I and 86Y. The degradation in image quality due to the imperfection of these isotopes is quantified for a human three-dimensional (3-D) positron emission tomography (PET) system. An acquisition protocol to obtain similar image quality as for 18F imaging is determined by Monte Carlo simulations. METHODS: The effects of larger positron range, associated singles and the other decay modes on image quality are determined by extensive Monte Carlo simulations of the Allegro scanner. Spatial resolution was evaluated for both isotopes and compared to spatial resolution of 18F. The loss in sensitivity due to triple coincidences was determined as a function of the axial acceptance angle of the PET scanner. The performance of the scanner at low count rates was studied by determining the noise equivalent count (NEC) values for different upper energy thresholds. The image degrading effect of spurious coincidences is taken into account by adding another factor to the NEC calculation. This allowed the contribution of spurious coincidences to be minimized by using a setting for the appropriate energy window. For this optimal energy window the amount of spurious and scattered coincidences was quantified. Simulations of count rate performance were also done to determine the peak NEC and the activity at which the maximum occurred. RESULTS: Spatial resolution degradation, compared to 18F, is about 0.5 mm for 86Y and 1 mm for 124I. Associated singles have a similar effect as scattered coincidences, as they also add a background to the image. The effect, however, is less important than the effect of scatter. The fraction of triple coincidences is quite small for a 3-D PET scanner used for humans as the axial acceptance angle is still moderate. For the Allegro with an energy resolution of 18% the optimal upper energy threshold was determined at 600 keV. For 124I this leads to 2.5% extra contamination that needs to be added to the scatter fraction. For 86Y this fraction is about 5.5%. CONCLUSION: 3-D PET images of 124I and 86Y have lower spatial resolution. For PET scanners used for humans the difference is not as important as for scanners used for animals. The limited positron decay fraction of both isotopes can be compensated by increasing the imaging time by a factor of 3-5 (same activity). A short coincidence window limits the contamination from other decay modes. Good energy resolution allows setting a selective upper energy threshold to limit the effect of spurious coincidences. With an appropriate setting of the energy window it should be possible to obtain good image quality in a relatively short time because of the high sensitivity of 3-D PET scanners. PMID- 16479244 TI - The effect of verapamil on the restoration of myocardial perfusion and functional recovery in patients with angiographic no-reflow after primary percutaneous coronary intervention. AB - OBJECTIVE: Angiographic thrombolysis in myocardial infarction (TIMI) flow grade < or = 2 after primary percutaneous coronary intervention (PCI), defined as angiographic no-reflow, predicts poor functional recovery in patients with acute myocardial infarction. We investigated the effect of verapamil on the restoration of myocardial perfusion and functional recovery in patients with angiographic no reflow after PCI. METHODS: 99mTc tetrofosmin single photon emission computed tomographic (SPECT) imaging was performed (before, immediately after and 1 month after PCI) in 101 consecutive patients with acute myocardial infarction. The defect score was calculated as the sum of perfusion defect in a 13-segment model (scores of 3, complete defect to 0, normal perfusion). The asynergic score, defined as the number of asynergic segments, was assessed by echocardiography before and 1 month later. Multiple logistic regression analysis was performed to elucidate the effect of verapamil administration. RESULTS: Of 101 patients, 32 (31%) had angiographic no-reflow and were divided into two groups: 18 patients with verapamil (group 1) and 14 patients without verapamil (group 2). Sixty-nine patients had TIMI grade 3 reflow after PCI (group 3). The change in the defect score 1 month after PCI in group 1 was significantly larger than that in group 2 (P = 0.003). The asynergic score improved more at 1 month in group 1 compared to that in group 2 (P = 0.007). Moreover, logistic regression analysis revealed that TIMI grade reflow < or = 2 after PCI (P = 0.04, OR = 5.51), the defect score before PCI (P = 0.03, OR = 1.15), the asynergic score before PCI (P = 0.01, OR = 0.64) and the administration of verapamil (P = 0.002, OR = 22.4) were independently associated with successful myocardial reperfusion immediately after PCI. CONCLUSIONS: Intracoronary verapamil restored myocardial perfusion in patients with angiographic no-reflow after PCI and lead to better functional recovery after acute myocardial infarction. PMID- 16479245 TI - Post-test quality control for the single blood sample technique in glomerular filtration rate measurement in children. AB - BACKGROUND: While the value of the single blood sample (SBS) method for estimating 51Cr-EDTA plasma clearance has been repeatedly demonstrated, some nuclear medicine physicians are still reluctant to use it because of the lack of quality control parameters. PURPOSE: To present a post-test quality control procedure for the SBS technique in children. METHODS: In addition to the SBS clearance calculated using the specific paediatric SBS method, three artificial slope intercept (ASI) method clearances were calculated by assuming the distribution volume as, respectively, 20%, 25% and 30% of body weight. By dividing the injected activity by the distributional volume, the initial plasma concentrations (A0,30%, A0,25% and A0,20%) were calculated. Using these A0 values and the available single sample, ASI clearances were calculated by using the classical slope-intercept method. The working hypothesis of this approach was as follows. In the absence of significant errors, the three ASI clearance values should be close to that of the SBS method. This hypothesis has been tested using both simulated and patients' data. RESULTS: The results of the simulated study showed that an error in the injected dose produced variable differences between SBS and ASI clearances depending on the clearance values. The effect of an error on the plasma sample also varied as a function of the clearance values. The analysis of patient data revealed that the ASI approach allowed the identification of patients in whom the classical slope-intercept method suggested the presence of a possible error. CONCLUSION: A post-test quality control procedure for the SBS GFR measurement is presented. When the SBS clearance shows a difference with the ASI method (> 10 ml . min(-1) per 1.73 m2), the presence of an error is highly probable. A smaller difference, however, does not exclude erroneous data. PMID- 16479246 TI - Radioiodine whole-body scans, thyroglobulin levels, 99mTc-MIBI scans and computed tomography: results in patients with lung metastases from differentiated thyroid cancer. AB - OBJECTIVES: The correlation between a 131I whole-body scan (WBS), a 99mTc sestamibi (99mTc-MIBI) WBS, a computed tomography (CT) scan and the value of routine follow-up for 131I WBS and thyroglobulin (Tg) levels in patients with lung metastases from differentiated thyroid cancer was assessed. METHOD: Pulmonary metastases were detected in 32 patients out of 583 with differentiated thyroid cancer (DTC) who were admitted to our clinic between 1985 and 2004 (age range, 22-79 years; mean, 58 +/- 19 years; 15 women and 17 men). Pulmonary metastases were diagnosed by considering the 131I WBS, increased Tg levels and/or other positive radiological findings. Papillary carcinoma was diagnosed in 15/32 patients and follicular carcinoma in 13/32. A mixed type found in 4/32 patients was classified histopathologically. A total of 3.7-53.65 GBq (100-1450 mCi) 131I was given to each patient. The duration of follow-up ranged from 36 to 240 months. A 131I WBS, the determination of Tg levels and/or a CT scan were carried out in the assessment of a diagnosis and follow-up of patients with lung metastases. A 99mTc-MIBI WBS was performed on 19 patients who were chosen at random from the 583. RESULTS: Nineteen of 32 patients had lung metastases before they received the first 131I treatment. Six of the 32 had distant-organ metastases other than in the lungs. Four of these six patients had only lung and bone metastases. Pulmonary metastases were observed on the 131I WBS patients 31/32 (96.8%) whereas no pulmonary metastases, were detected on the CT scans in 3/32 patients. The last diagnostic whole-body scan (DWBS) was normal in 13/32 patients. At the first examination, the Tg levels in 27/32 (84.4%) patients were below 30 ng . ml(-1). At the final examination, 20/32 (62.5%) patients had Tg levels higher than 30 ng . ml(-1), while Tg levels were lower than 30 ng . ml(-1) in 12/32 patients. Tg levels decreased in 21/32 and increased in 3/32 patients. The 131I WBS continued to be abnormal in 2/3 patients with increased Tg levels but became normal in one patient whose CT scan still showed macro-nodular lesions. Tg levels did not change significantly in 8/32 patients. The 131I WBS became normal in 5/8 patients, while the CT scans for 4/5 showed micro-nodules. Metastases were detected in 12/19 patients who underwent 99mTc-MIBI whole-body scanning: 18/19 showed metastases on the 131I WBSs and 17/19 on the CT scans. Of the seven patients without a sign of metastasis on the 99mTc-MIBI WBS, one was negative in terms of metastasis on the 131I WBS and one on the CT scan. Fibrosis was observed on the CT scans of 2/32 patients. One patient developed dedifferentiation, as determined by the negative 131I WBS and positive CT scan. CONCLUSION: 131I whole-body scanning and the determination of Tg levels are the most important procedures for the evaluation of lung metastases in differentiated thyroid cancer. Computed tomography is a useful addition to 131I whole-body scanning. MIBI imaging alone may not be enough to detect lung metastases from differentiated thyroid cancer. PMID- 16479247 TI - Striatal dopamine transporter density in drug naive patients with attention deficit/hyperactivity disorder. AB - BACKGROUND AND AIM: Dopamine transporters are the target of psychostimulants used for treatment of attention-deficit/hyperactivity disorder (ADHD). Therefore, the present study aimed to evaluate striatal dopamine transporter density in adult patients with ADHD. METHODS: Twenty patients (11 female, nine male; mean age 35+/ 7 years) and 20 control subjects (11 female, nine male, mean age 32 +/- 8 years) were examined with SPECT using the specific radiotracer 123I-FP-CIT. The ratio of striatal to cortical radioactivity concentration was used for semiquantitative evaluation of dopamine transporter binding potential (V3''). There was a significant influence of age (P < 0.001) and a trend towards an influence of gender (P = 0.053) on V3''. An ANCOVA with these covariates showed a slightly higher V3'' in the patients than in the control subjects (4.24 +/- 0.48 vs. 4.03 +/- 0.56; P = 0.02). CONCLUSION: This study provides further in-vivo evidence for an involvement of the dopamine transporter in ADHD. However, compared to previous studies, the increase of dopamine transporter density in the patient group is less pronounced here. PMID- 16479248 TI - Preparation of 99mTc-labelled conjugates of ouabagenin and their biological evaluation in animal models. AB - BACKGROUND: Ouabagenin and its 1,19-acetonide were conjugated with nitrilotriacetic acid (NTA) and diethylenetriaminepentaacetic acid (DTPA) through their respective anhydrides. METHODS: The reaction mixtures were exhaustively purified by silica gel column chromatography and preparative high-performance liquid chromatography to furnish the ligands in good purity and moderate yield. These ligands were labelled with 99mTc to produce four chelates in 90-95% yield. Of these chelates the 99mTc-oubagenin-NTA conjugate and the corresponding acetonide exhibited appreciable myocardial uptake with respect to that of other vicinal organs in a guinea-pig model. However, all these 99mTc chelates exhibited poor heart-to-blood ratios, which could be attributed to the absence of a 3beta sugar residue in this molecule. CONCLUSION: The result is in agreement with that previously reported in connection with radioiodinated digoxin and digoxigenin derivatives. PMID- 16479251 TI - Autologous transplantation of rabbit limbal epithelia cultured on fibrin gels for ocular surface reconstruction. AB - PURPOSE: Regeneration of the corneal epithelium could be severely impaired in patients suffering from limbal stem cell deficiency. The purpose of this study was to evaluate the restoration of the corneal epithelium by grafting onto denuded corneas autologous limbal cells cultured on fibrin gels. The rabbit model was chosen to allow the microscopic evaluation over time after grafting. METHODS: Rabbit limbal epithelial cells (RLECs) were isolated and cultured from small limbal biopsies (3 mm2). The epithelium was separated from stroma after dispase digestion and put in culture on lethally irradiated fibroblasts used as a feeder layer. At the first passage, RLECs were cultured on a fibrin gel matrix. At confluence, the cultured epithelia were grafted in vivo on denuded autologous rabbit corneas. At different postoperative times, grafted and control (without graft or grafted with fibrin gels only) rabbit corneas were compared in vivo with a slit lamp microscope, and in situ by histological and immunohistological microscopy of harvested biopsies. RESULTS: A small limbal biopsy was sufficient to generate enough RLECs to prepare several grafts and to perform cell analysis. Only two weeks were required to produce a cultured epithelium suitable for autologous transplantation. One month after grafting, a normal corneal phenotype was observed on the ocular surface of grafted rabbits in contrast to the control rabbits (ungrafted or grafted with fibrin gel only) where histological signs of conjunctivalization were found. The absence of goblet cells and negative staining for keratin 4 confirmed that the cultured cells persisted and that the epithelium regenerated after grafting was not from conjunctival origin. CONCLUSIONS: Our results demonstrate that an autologous epithelium cultured on a physiologically biodegradable matrix can be prepared from a small biopsy and grafted on denuded cornea. The autologous graft allows epithelial regeneration from cultured cells and promotes corneal healing of unilateral total stem cell deficiency. PMID- 16479253 TI - Evaluation of an inflammation-based prognostic score in patients with inoperable gastro-oesophageal cancer. AB - There is increasing evidence that the presence of an ongoing systemic inflammatory response is associated with poor outcome in patients with advanced cancer. The aim of the present study was to examine whether an inflammation-based prognostic score (Glasgow Prognostic score, GPS) was associated with survival, in patients with inoperable gastro-oesophageal cancer. Patients diagnosed with inoperable gastro-oesophageal carcinoma and who had measurement of albumin and C reactive protein concentrations, at the time of diagnosis, were studied (n=258). Clinical information was obtained from a gastro-oesophageal cancer database and analysis of the case notes. Patients with both an elevated C-reactive protein (>10 mg l(-1)) and hypoalbuminaemia (<35 g l(-1)) were allocated a GPS score of 2. Patients in whom only one of these biochemical abnormalities was present were allocated a GPS score of 1, and patients with a normal C-reactive protein and albumin were allocated a score of 0. On multivariate survival analysis, age (hazard ratio (HR) 1.22, 95% CI 1.02-1.46, P<0.05), stage (HR 1.55, 95% CI 1.30 1.83, P<0.001), the GPS (HR 1.51, 95% CI 1.22-1.86, P<0.001) and treatment (HR 2.53, 95% CI 1.80-3.56, P<0.001) were significant independent predictors of cancer survival. A 12-month cancer-specific survival in patients with stage I/II disease receiving active treatment was 67 and 60% for a GPS of 0 and 1, respectively. For stage III/IV disease, 12 months cancer-specific survival was 57, 25 and 12% for a GPS of 0, 1 and 2, respectively. In the present study, the GPS predicted cancer-specific survival, independent of stage and treatment received, in patients with inoperable gastro-oesophageal cancer. Moreover, the GPS may be used in combination with conventional staging techniques to improve the prediction of survival in patients with inoperable gastro-oesophageal cancer. PMID- 16479252 TI - Induction of vitronectin and integrin alphav in the retina after optic nerve injury. AB - PURPOSE: Vitronectin is a secreted glycoprotein present in blood plasma and is present in the extracellular matrix of many tissues. It was found in the retinal cDNA library that contains genes whose expression is upregulated after optic nerve injury in a previous study. The purpose of this study was to assess the temporal and spatial changes in expression of vitronectin and integrin alphav in the retina following optic nerve injury. METHODS: Adult Balb/c mice underwent crush of the optic nerve in one eye only. RT-PCR was used to determine the temporal expression of vitronectin mRNA in the retina after injury. In addition, expression at the protein level in the retina and the optic nerve of vitronectin and its major receptor subunit, integrin alphav, was analyzed by immunohistochemistry. RESULTS: Upregulation of vitronectin mRNA in the retina was detected at one day after injury, peaking at three days, and maintained up to one week. An elevated expression of vitronectin protein was also observed in the inner retina, optic nerve head, and the optic nerve after nerve crush. In the inner retina, the increased expression of vitronectin was found in retinal ganglion cells (RGCs) and its surrounding extracellular matrix. Expression of integrin alphav was also increased in the RGC layer and in the glial cells of the nerve head and the crush site. CONCLUSIONS: As vitronectin is an extracellular protein that can support cell attachment and promote neurite extension, elevated expression of vitronectin and its receptor may facilitate axonal regeneration following injury. We propose that treatment sustaining secretion of endogenous vitronectin or direct application of exogenous vitronectin may be a method to augment regeneration of the severed optic axons. PMID- 16479254 TI - Tissue microarray analysis of human FRAT1 expression and its correlation with the subcellular localisation of beta-catenin in ovarian tumours. AB - The mechanisms involved in the pathogenesis of ovarian cancer are poorly understood, but evidence suggests that aberrant activation of Wnt/beta-catenin signalling pathway plays a significant role in this malignancy. However, the molecular defects that contribute to the activation of this pathway have not been elucidated. Frequently rearranged in advanced T-cell lymphomas-1 (FRAT1) is a candidate for the regulation of cytoplasmic beta-catenin. In this study, we developed in situ hybridisation probes to evaluate the presence of FRAT1 and used an anti-beta-catenin antibody to evaluate by immunohistochemistry the expression levels and subcellular localisation of beta-catenin in ovarian cancer tissue microarrays. Expression of FRAT1 was found in some human normal tissues and 47% of ovarian adenocarcinomas. A total of 46% of ovarian serous adenocarcinomas were positive for FRAT1 expression. Accumulation of beta-catenin in the nucleus and/or cytoplasm was observed in 55% ovarian adenocarcinomas and in 59% of serous adenocarcinomas. A significant association was observed in ovarian serous adenocarcinomas between FRAT1 and beta-catenin expression (P<0.01). These findings support that Wnt/beta-catenin signalling may be aberrantly activated through FRAT1 overexpression in ovarian serous adenocarcinomas. The mechanism behind the overexpression of FRAT1 in ovarian serous adenocarcinomas and its significance is yet to be investigated. PMID- 16479255 TI - Expression of Y-box-binding protein dbpC/contrin, a potentially new cancer/testis antigen. AB - Y-box-binding proteins are members of the human cold-shock domain protein superfamily, which includes dbpA, dbpB/YB-1, and dbpC/contrin. dbpC/contrin is a germ cell-specific Y-box-binding protein and is suggested to function as a nuclear transcription factor and RNA-binding protein in the cytoplasm. Whereas ubiquitous dbpB/YB-1 expression has been well studied in various types of human carcinomas as a prognostic or predictive marker, the dbpC/contrin expression in human tumour cells has not been reported. In this report, we provide the first evidence showing that dbpC was highly expressed in human testicular seminoma and ovarian dysgerminomas, and in carcinomas in other tissues and that its expression in normal tissues is nearly restricted to germ cells and placental trophoblasts. These results indicate that dbpC/contrin would be a potentially novel cancer/testis antigen. PMID- 16479256 TI - Ubiquitous mitochondrial creatine kinase downregulated in oral squamous cell carcinoma. AB - In this study, we performed two-dimensional electrophoresis (2-DE) and matrix assisted laser desorption/ionisation time of fly mass spectrometry to identify the protein(s) associated with the development of oral squamous cell carcinomas (OSCCs) by comparing patterns of OSCC-derived cell lines with normal oral keratinocytes (NOKs), and found that downregulation of ubiquitous mitochondrial creatine kinase (CKMT1) could be a good candidate. Decreased levels of CKMT1 mRNA and protein were detected in all OSCC-derived cell lines examined (n=9) when compared to those in primary normal oral keratinocytes. Although no sequence variation in the coding region of the CKMT1 gene with the exception of a nonsense mutation in exon 8 was identified in these cell lines, we found a frequent hypermethylation in the CpG island region. CKMT1 expression was restored by experimental demethylation. In addition, when we transfected CKMT1 into the cell lines, they showed an apoptotic phenotype but no invasiveness. In clinical samples, high frequencies of CKMT1 downregulation were detected by immunohistochemistry (19 of 52 (37%)) and quantitative real-time RT-PCR (21 of 50 (42%)). Furthermore, the CKMT1 expression status was significantly correlated with tumour differentiation (P<0.0001). These results suggest that the CKMT1 gene is frequently inactivated during oral carcinogenesis and that an epigenetic mechanism may regulate loss of expression, which may lead to block apoptosis. PMID- 16479257 TI - Epigenetic inactivation of TCF2 in ovarian cancer and various cancer cell lines. AB - Transcription factor 2 gene (TCF2) encodes hepatocyte nuclear factor 1beta (HNF1beta), a transcription factor associated with development and metabolism. Mutation of TCF2 has been observed in renal cell cancer, and by screening aberrantly methylated genes, we have now identified TCF2 as a target for epigenetic inactivation in ovarian cancer. TCF2 was methylated in 53% of ovarian cancer cell lines and 26% of primary ovarian cancers, resulting in loss of the gene's expression. TCF2 expression was restored by treating cells with a methyltransferase inhibitor, 5-aza-2'deoxycitidine (5-aza-dC). In addition, chromatin immunoprecipitation showed deacetylation of histone H3 in methylated cells and, when combined with 5-aza-dC, the histone deacetylase inhibitor trichostatin A synergistically induced TCF2 expression. Epigenetic inactivation of TCF2 was also seen in colorectal, gastric and pancreatic cell lines, suggesting general involvement of epigenetic inactivation of TCF2 in tumorigenesis. Restoration of TCF2 expression induced expression of HNF4alpha, a transcriptional target of HNF1beta, indicating that epigenetic silencing of TCF2 leads to alteration of the hepatocyte nuclear factor network in tumours. These results suggest that TCF2 is involved in the development of ovarian cancers and may represent a useful target for their detection and treatment. PMID- 16479258 TI - Spatial and space-time clustering of childhood acute leukaemia in France from 1990 to 2000: a nationwide study. AB - This study aimed to investigate the spatial and space-time distributions of cases of childhood acute leukaemia (CL) during 1990-2000 over the whole French territory. A global spatial heterogeneity and a spatial autocorrelation were first considered using the methods proposed by Potthoff and Whittinghill, Moran and Rogerson methods. The presence of space-time interaction between the places of residence and the dates of diagnosis was investigated with the Knox's test. Finally, the Kulldorff's statistic permitted to scan the whole territory in search for localised clusters. Two time periods were considered (1990-1994, 1995 2000). Overall, a statistically significant spatial heterogeneity of a very small magnitude was observed in the incidence of CL over 1990-1994, but neither over 1995-2000 nor over the whole time period. Moreover, a significant overdispersion of 5.5% was evidenced for 0-4 year children living in isolated areas with more than 50 inhabitants per km(2). Cases older than 10 years living in the same area at diagnosis also tended to cluster within 6 months. PMID- 16479259 TI - Phenotypic and genotypic heterogeneity in the Lynch syndrome: diagnostic, surveillance and management implications. AB - Lynch syndrome is the most common form of hereditary colorectal cancer (CRC). This review covers the cardinal features of Lynch syndrome with particular emphasis upon its diagnostic criteria, molecular genetics, natural history, genetic counseling, surveillance and management. Considerable attention has been given to the etiologic role of mismatch repair (MMR) genes as well as low penetrance alleles and modifier genes. The American founder mutation, a deletion of exons 1-6 of MSH2, is discussed in some detail, owing to its high frequency in the US (19 000-30 000 carriers). Genetic counseling is essential prior to patients' undergoing DNA testing and again when receiving their test results. Families with a lower incidence of CRC and extracolonic cancers, in the face of being positive for Amsterdam I criteria but who do not have MMR deficiency by tumor testing, are probably not Lynch syndrome, and thereby should preferably be designated as familial CRC of undetermined type. Patients who are either noncompliant or poorly compliant with colonoscopy, and who are MMR mutation positive, may be candidates for prophylactic colectomy, while MMR mutation positive women who are noncompliant with gynecologic surveillance may be candidates for prophylactic hysterectomy and bilateral salpingo-oophorectomy. PMID- 16479260 TI - An utter refutation of the "fundamental theorem of the HapMap". AB - The International HapMap Project was proposed in order to quantify linkage disequilibrium (LD) relationships among human DNA polymorphisms in an assortment of populations, in order to facilitate the process of selecting a minimal set of markers that could capture most of the signal from the untyped markers in a genome-wide association study. The central dogma can be summarized by the argument that if a marker is in tight LD with a polymorphism that directly impacts disease risk, as measured by the metric r(2), then one would be able to detect an association between the marker and disease with sample size that was increased by a factor of 1/r(2) over that needed to detect the effect of the functional variant directly. This "fundamental theorem" holds, however, only if one assumes that the LD between loci and the etiological effect of the functional variant are independent of each other, that they are statistically independent of all other etiological factors (in exposure and action), that sampling is prospective, and that the estimates of r(2) are accurate. None of these are standard operating assumptions, however. We describe the ramifications of these implicit assumptions, and provide simple examples in which the effects of a functional variant could be unequivocally detected if it were directly genotyped, even as markers in high LD with the functional variant would never show association with disease, even in infinite sample sizes. Both theoretical and empirical refutation of the central dogma of genome-wide association studies is thus presented. PMID- 16479261 TI - It's "back to school" for genetic screening. AB - Implementation of population genetic screening programmes requires consideration of strategies for reaching the greatest proportion of the target population in order to achieve maximum awareness. This article reviews the current strategy of school-based population genetic screening programmes. The school environment is an ideal setting for offering relevant genetic screening programmes as it provides an opportunity to engage people at a time when they are exposed to a range of educational experiences and are sufficiently mature to be involved in decision-making processes. Such programmes allow all students, not only those studying biology, an opportunity to be educated and experience genetic screening in a supportive environment, ultimately increasing understanding and empowering students. While the major form of genetic screening in schools has been for reproductive health information (eg carrier screening for TaySachs disease and cystic fibrosis), genetic screening in schools for other conditions may be a timely proposition. PMID- 16479262 TI - Molecular characterization of a novel PAX9 missense mutation causing posterior tooth agenesis. AB - Autosomal dominant mutations in the gene encoding the paired box containing transcription factor PAX9 are associated with nonsyndromic human tooth agenesis that primarily affect posterior dentition. The molecular mechanisms contributing to its pathogenesis are poorly understood. In this study, we describe a novel mutation in PAX9 in a family with molar oligodontia. This heterozygous mutation results in the substitution of a highly conserved isoleucine residue by phenylalanine within the carboxyl-terminal subdomain of the paired domain. Immunolocalization and cell fractionation studies to ascertain the subcellular localization of the Ile87Phe protein showed that both wild-type and mutant proteins are synthesized in mammalian cells and that the mutation does not alter the nuclear localization of the mutant protein. Gel-shift assays using two cognate paired-domain recognition sequences, e5 and CD19-2(A-ins), revealed that while wild-type Pax9 binds to both sequences, the mutant protein was unable to bind these sites. In addition, the latter did not alter the DNA-binding activities of wild-type Pax9. Furthermore, we evaluated the ability of the Ile87Phe mutant protein to form a complex with a partner protein, Msx1, and found that the mutation under study has no effect on this interaction. Based on our observed defects in DNA binding by the mutant protein, we propose a loss-of function mechanism that contributes to haploinsufficiency of PAX9 in this family with posterior tooth agenesis. PMID- 16479265 TI - On the dissociation and conformation of gas-phase methonium ions. AB - The dissociation pathways of both doubly and singly charged methonium ions, (CH(3))(3)N(+) -(CH(2))(n)-(+)N(CH(3))(3).X(-) (n = 6,10; X = Br, I, and OAc), are measured using blackbody infrared radiative dissociation (BIRD) and SORI-CAD in a Fourier transform mass spectrometer. SORI-CAD of the doubly charged decamethonium ions results primarily in the formation of even electron ions by hydrogen rearrangements. In contrast, homolytic bond cleavage to form two odd electron ions is highly favored in the hexamethonium ion, presumably due to increased Coulomb repulsion in this ion. For BIRD of the singly charged salts, ions are mass selected and dissociated by heating the vacuum chamber to elevated temperatures. Under the low pressure conditions of our experiment, energy is transferred from the chamber walls to the ions by the absorption of blackbody radiation. From the temperature dependence of the unimolecular rate constants for dissociation, Arrhenius activation energies in the zero-pressure limit are obtained. The primary dissociation pathways correspond to counterion substitution reactions which result in loss of N(CH(3))(3) and CH(3)X. For hexamethonium and decamethonium with X = Br or I, the branching ratios for these pathways differ dramatically; the ratio of loss of N(CH(3))(3) and CH(3)Br is 3.8 and 0.4 for hexamethonium and decamethonium bromide, respectively. The hexamethonium acetate salt has a branching ratio of 0.1. The Arrhenius activation energies for hexamethonium (Br or I) and decamethonium (Br or I) are 0.9 and 1.0 eV, respectively. This value for hexamethonium acetate is 0.6 eV. Molecular dynamics simulations and Monte Carlo conformation searching are used to obtain the lowest energy structures of hexamethonium and decamethonium bromide. These calculations indicate that the methonium ion folds around the counterion to form a cyclic salt bridge structure in which both quaternary nitrogens interact with the oppositely charged counterion. The significantly different branching ratios observed for these ions is attributed to the large change in orientation of the counterion with respect to the ammonium centers as the number of methylene groups in these ions increases. Similar ion conformational differences appear to explain the fragmentation for the OAc counter ion as well. PMID- 16479266 TI - The Times of Ira Hirsh: Multiple Ranges of Auditory Temporal Perception. AB - Ira Hirsh was among the first to recognize that the auditory system does not deal with temporal information in a unitary way across the continuum of time intervals involved in speech processing. He identified the short range (extending from 1 to 20 milliseconds) as that of phase perception, the range between 20 and 100 milliseconds as that in which auditory patterns emerge, and the long range from 100 milliseconds and longer as that of separate auditory events. Furthermore, he also was among the first to recognize that auditory time perception heavily depended on spectral context. A study of the perception of sequences representing different temporal orders of three tones, by Hirsh and the author (e.g., Divenyi and Hirsh, 1978) demonstrated the dependence of auditory sequence perception on both time range and spectral context, and provided a bridge between Hirsh's view of auditory time and Bregman's view of stream segregation. A subsequent search by the author for psychophysical underpinnings of the cocktail-party phenomenon (e.g., Divenyi and Haupt, 1997) suggests that segregation of simultaneous streams of speech might rely on the ability to follow spectral changes in the demisyllabic-to-syllabic (100 to 200 milliseconds) range (i.e., Hirsh's long range).Learning Outcomes: As a result of this activity, the participant will be able to (1) describe the importance of temporal processing in hearing; and (2) identify time ranges where the auditory system will spontaneously adopt different analysis techniques. PMID- 16479267 TI - Is arginine a zwitterion in the gas phase? PMID- 16479268 TI - Hydration energies of divalent metal ions, Ca2+ (H2O)n (N = 5-7) and Ni2+ (H2O)m (N = 6-8), obtained by blackbody infrared radiative dissociation. PMID- 16479269 TI - Dissociation of heme-globin complexes by blackbody infrared radiative dissociation: molecular specificity in the gas phase? AB - The temperature dependence of the unimolecular kinetics for dissociation of the heme group from holo-myoglobin (Mb) and holo-hemoglobin alpha-chain (Hb-alpha) was investigated with blackbody infrared radiative dissociation (BIRD). The rate constant for dissociation of the 9 + charge state of Mb formed by electrospray ionization from a "pseudo-native" solution is 60% lower than that of Hb-alpha at each of the temperatures investigated. In solutions of pH 5.5-8.0, the thermal dissociation rate for Mb is also lower than that of HB-alpha (Hargrove, M. S. et al. J. Biol. Chem.1994, 269, 4207-4214). Thus, Mb is thermally more stable with respect to heme loss than Hb-alpha both in the gas phase and in solution. The Arrhenius activation parameters for both dissociation processes are indistinguishable within the current experimental error (activation energy 0.9 eV and pre-exponential factor of 10(8-10) s(-1)). The 9+ to 12+ charge states of Mb have similar Arrhenius parameters when these ions are formed from pseudo-native solutions. In contrast, the activation energies and pre-exponential factors decrease from 0.8 to 0.3 eV and 10(7) to 10(2) s(-1), respectively, for the 9 + to 12 + charge states formed from acidified solutions in which at least 50% of the secondary structure is lost. These results demonstrate that gas-phase Mb ions retain clear memory of the composition of the solution from which they are formed and that these differences can be probed by BIRD. PMID- 16479272 TI - Identification and properties of microsatellite markers in tsetse flies Glossina morsitans sensu lato (Diptera: Glossinidae). AB - Genomic libraries enriched for simple sequence repeats were constructed for Glossina morsitans morsitans, G. m. submorsitans, and G. m. centralis. Sixteen microsatellite markers were isolated from the libraries and evaluated on flies from natural G. m. morsitans populations and other Glossina species in the Morsitans and Palpalis species groups. The primers amplified appropriate sized DNA fragments in the Morsitans and Palpalis groups. In G. morsitans s.l., eight of 12 dinucleotide repeats and four of 12 trinucleotide repeats were polymorphic. The polymorphic loci showed a mean 7.5 +/- 4.8 alleles per locus and their mean heterozygosity was 55.8 +/- 7.7%. PMID- 16479273 TI - Timely opportunities for families and their healthcare providers: a commentary on Pyle et al. (2005). AB - This commentary on S. A. Pyle, C. K. Haddock, N. Hymowitz, J. Schwab, and S. Meshberg (2005) addresses the complex nature of establishing family rules about exposure to tobacco smoke (ETS). Particular attention is paid to how family rules are established and the challenges in enforcing rules among adult family members. Findings are also discussed in terms of the limits of family rule making and the need for negotiation and conflict resolution. The compounding influence of socioeconomic status and parental education in monitoring ETS deserves serious attention in future research. Healthcare providers are urged to discuss family rules with their patients with an eye toward preventative interventions. PMID- 16479275 TI - Structure of cationized arginine (arg.m, m = h, li, na, k, rb, and cs) in the gas phase: further evidence for zwitterionic arginine. AB - The gas-phase structures of cationized arginine, Arg.M(+), M = Li, Na, K, Rb, and Cs, were studied both by hybrid method density functional theory calculations and experimentally using low-energy collisionally activated and thermal radiative dissociation. Calculations at the B3LYP/LACVP++** level of theory show that the salt-bridge structures in which the arginine is a zwitterion (protonated side chain, deprotonated C-terminus) become more stable than the charge-solvated structures with increasing metal ion size. The difference in energy between the most stable charge-solvated structure and salt-bridge structure of Arg.M(+) increases from -0.7 kcal/mol for Arg.Li(+) to +3.3 kcal/mol for Arg.Cs(+). The stabilities of the salt-bridge and charge-solvated structures reverse between M = Li and Na. These calculations are in good agreement with the results of dissociation experiments. The low-energy dissociation pathways depend on the cation size. Arginine complexed with small cations (Li and Na) loses H(2)O, while arginine complexed with larger cations (K, Rb, and Cs) loses NH(3). Loss of H(2)O must come from a charge-solvated ion, whereas the loss of NH(3) can come from the protonated side chain of a salt-bridge structure. The results of dissociation experiments using several cationized arginine derivatives are consistent with the existence of these two distinct structures. In particular, arginine methyl esters, which cannot form salt bridges, dissociate by loss of methanol, analogous to loss of H(2)O from Arg.M(+); no loss of NH(3) is observed. Although dissociation experiments probe gas-phase structure indirectly, the observed fragmentation pathways are in good agreement with the calculated lowest energy isomers. The combination of the results from experiment and theory provides strong evidence that the structure of arginine-alkali metal ion complexes in the gas phase changes from a charge-solvated structure to a salt-bridge structure as the size of the metal ion increases. PMID- 16479276 TI - Amino acid side chain interactions in the presence of salts. AB - The effects of salt on the intermolecular interactions between polar/charged amino acids are investigated through molecular dynamics simulations. The mean forces and associated potentials are calculated for NaCl salt in the 0-2 M concentration range at 298 K. It is found that the addition of salt may stabilize or destabilize the interactions, depending on the nature of the interacting molecules. The degree of (de)stabilization is quantified, and the origin of the salt-dependent modulation is discussed based upon an analysis of solvent density profiles. To gain insight into the molecular origin of the salt modulation, spatial distribution functions (sdf's) are calculated, revealing a high degree of solvent structuredness in all cases. The peaks in the sdf's are consistent with long-range hydrogen-bonding networks connecting the solute hydrophilic groups, and that contribute to their intermolecular solvent-induced forces. The restructuring of water around the solutes as they dissociate from close contact is analyzed. This analysis offers clues on how the solvent structure modulates the effective intermolecular interactions in complex solutes. This modulation results from a critical balance between bulk electrostatic forces and those exerted by (i) the water molecules in the structured region between the monomers, which is disrupted by ions that transiently enter the hydration shells, and (ii) the ions in the hydration shells in direct interactions with the solutes. The implications of these findings in protein/ligand (noncovalent) association/dissociation mechanisms are briefly discussed. PMID- 16479277 TI - Exploring protein energy landscapes with hierarchical clustering. AB - In this work we present a new method for investigating local energy minima on a protein energy landscape. The CABS (CAlpha, CBeta and the center of mass of the Side chain) method was employed for generating protein models, but any other method could be used instead. Calpha traces from an ensemble of models are hierarchical clustered with the HCPM (Hierarchical Clustering of Protein Models) method. The efficiency of this method for sampling and analyzing energy landscapes is shown. PMID- 16479280 TI - The ying and yang of asymmetric aminocatalysis. AB - During the last six years the asymmetric catalysis of carbonyl transformations via iminium ion and enamine intermediates using chiral amines as organocatalysts has grown most remarkably. In this personal account an overview of this area is given. The field can be divided into two sub areas: (a) Iminium catalysis, which is typically used for cycloadditions and conjugate additions to enals and enones and (b) Enamine catalysis, which is commonly used in electrophilic alpha substitution reactions of ketones and aldehydes. A common origin of the two catalysis principles is proposed and their recent merger in tandem sequences is discussed. PMID- 16479281 TI - Carrier enabled catalytic reaction cascades. AB - Catalytic cascade reactions increase atom efficiency and help to reduce waste. Therefore they have recently attracted considerable attention. When performing them we have experienced that the immobilisation of the catalyst and the nature of the support are crucial factors. The carriers can play various roles from facilitating filtration and compartmentalisation to specifically binding unwanted compounds in the reaction media. In this Feature Article we give an overview of our work on such carrier dependent cascade reactions. PMID- 16479282 TI - Intramolecular bromo-amination of 1,4-cyclohexadiene aminal: one-pot discrimination of two olefins and concise asymmetric synthesis of (-)-gamma lycorane. AB - The reaction of cyclohexa-2,5-dienyl-1-methylaldehyde and optically pure 1,2 diaryl-1,2-diamine followed by intramolecular bromo-amination produced a one-pot discrimination of two olefins in the cyclohexane system, which was used for the asymmetric synthesis of (-)-gamma-lycorane. PMID- 16479283 TI - Thioketone spin traps as mediating agents for free radical polymerization processes. AB - Thioketones are demonstrated to be suitable agents for controlling free radical polymerization processes: the polymerizations carry (pseudo) living characteristics indicating that the control process is induced by a persistent radical effect. PMID- 16479284 TI - An experimental proof for negative oxidation states of platinum: ESCA measurements on barium platinides. AB - ESCA-measurements on barium platinides provide the first spectroscopic evidence for negative oxidation states of platinum and are in excellent agreement with theoretical predictions based on quantum-chemical calculations. PMID- 16479285 TI - A simple and practical phase-separation approach to the recycling of a homogeneous metathesis catalyst. AB - The air stable asarone-derived Ru carbene 16, a robust olefin metathesis catalyst, can be easily separated after reaction by deposition on silica gel and reused up to nine times. This procedure provides products of excellent purity with low Ru content. PMID- 16479286 TI - Regioselective and enantiospecific rhodium-catalyzed allylic amination with thymine: synthesis of a new conformationally rigid nucleoside. AB - The regioselective and enantiospecific rhodium-catalyzed allylic amination of secondary allylic carbonates with N3-benzoyl thymine in conjunction with a stereoselective free radical cyclization provides a convenient method for the construction of a new conformationally rigid nucleoside. PMID- 16479287 TI - Conformal coating of nanoscale features of microporous Anodisc membranes with zirconium and titanium oxides. AB - We have successfully coated a nanofeatured material with ZrO2 and TiO2 using a polymer assisted deposition technique. PMID- 16479288 TI - Effective chiral recognition among ions in polar media. AB - Effective homochiral recognition occurs between the cationic [Fe(eilatin)3]2+ complex and TRISPHAT anions even in polar media such as 90% acetone-CHCl3 (de > or = 89%). PMID- 16479289 TI - Uranyl ion complexation by citric and tricarballylic acids: hydrothermal synthesis and structure of two- and three-dimensional uranium-organic frameworks. AB - The first crystal structure of uranyl citrate is reported, as well as that of uranyl sodium tricarballylate; both compounds are polymeric, with all acid groups coordinated, but they differ strongly in their coordination modes; the resulting assembly architecture is either three-dimensional with [(UO2)2(Hcit)2]2- metallacycle subunits with citrate or two-dimensional with tricarballylate. PMID- 16479290 TI - Uranium(IV) complexes of calix[n]arenes (n = 4, 6 and 8). AB - Reaction of UCl4 with calix[n]arenes (n = 4, 6 and 8) in THF or pyridine gave the mononuclear [UCl2(calix[4]arene--2H)(THF)2], bis-binuclear [U2Cl2(calix[6]arene- 6H)(THF)3]2 and trinuclear [Hpy]6[U3Cl11(calix[8]arene--7H)] complexes, respectively, which are the first U(IV) complexes of O-unsubstituted calixarenes. PMID- 16479291 TI - Trapping of a tert-adamantyl peroxyl radical in the gas phase. AB - A bridgehead adamantyl peroxyl radical has been prepared and isolated in the gas phase by the reaction of a distonic radical anion with dioxygen in a quadrupole ion-trap mass spectrometer. PMID- 16479292 TI - Application of the S(2infinity) and C(infinity) point groups for the prediction of polymer chirality. AB - Polymer chirality assignment is achieved with the first chemical applications of the S2infinity and C(infinity) molecular point groups to infinite cyclic polymers, obviating the usual dependence on translational symmetry operations. PMID- 16479293 TI - Baker's yeast-mediated enantioselective reduction of substituted fluorenones. AB - In the presence of DMSO as co-solvent and under vigorous agitation, baker's yeast in water was found to reduce substituted fluorenones to the corresponding fluorenols in good to excellent enantioselectivities. PMID- 16479294 TI - "Beta-cis-SAr effect" on decarbonylation from alpha,beta-unsaturated acyl and aroyl complexes. AB - Lone pair of heteroatom located at the beta-cis position in alpha,beta unsaturated acyl and aroyl group 10 metal complexes dramatically facilitated the stoichiometric and catalytic decarbonylation reactions. PMID- 16479295 TI - Poly[3-(5-octyl-thienylene-vinyl)-thiophene]: a side-chain conjugated polymer with very broad absorption band. AB - A novel polythiophene derivative, poly[3-(5-octyl-thienylene-vinyl)-thiophene] (POTVT) with conjugated thienylene vinyl side-chain, was synthesized, and the POTVT film shows a very broad absorption band-width covering from 300 nm to 700 nm after thermal annealing at 130 degrees C for 10 min. PMID- 16479296 TI - A bis(phosphinimino)methanide lanthanum amide as catalyst for the hydroamination/cyclisation, hydrosilylation and sequential hydroamination/hydrosilylation catalysis. AB - [La[N(SiHMe2)2]2[CH(PPh2NSiMe3)2]], which was obtained via an amine elimination starting from [CH2(PPh2NSiMe3)2] and [La[N(SiHMe2)2]3(THF)2], was used as catalyst for the hydroamination/cyclisation, the hydrosilylation and the sequential hydroamination/hydrosilylation reaction. PMID- 16479297 TI - Kinetic competition in flexible dye sensitised solar cells employing a series of polymer electrolytes. AB - Transient absorption spectroscopy is employed to study electron transfer dynamics in dye sensitised solar cells employing a series of polymer electrolytes, and correlated with device current-voltage characteristics. PMID- 16479298 TI - A covalently bonded AlQ3/SiO2 hybrid material with blue light emission by a conventional sol-gel approach. AB - We report the design and synthesis of a covalently bonded AlQ3/SiO2 hybrid material with strong blue light emission by a sol-gel approach, which make AlQ3 solution-processable and chemically stable. PMID- 16479299 TI - Generation of hierarchical pore systems in the titanosilicate ETS-10 by hydrogen peroxide treatment under microwave irradiation. AB - Supermicropores and well-defined mesopores with an average size of 10 nm were created in ETS-10 structure by post-synthesis treatment with H2O2 under microwave irradiation. Macropores were also formed and the external surface area of the material was increased during the treatment. PMID- 16479300 TI - Synthesis of a soluble poly(fluorenone). AB - The synthesis and characterisation of a soluble poly(fluorenone) is presented, a polymer with high electron affinity with potential for use in plastic electronic devices as an n-type material. PMID- 16479301 TI - Desorption electrospray ionization and electrosonic spray ionization for solid- and solution-phase analysis of industrial polymers. AB - Desorption electrospray ionization (DESI) and electrosonic spray ionization (ESSI), two new techniques, are used to measure average molecular weights and molecular weight distributions of solid-phase and solution-phase samples of the same polymers. PMID- 16479303 TI - Palladium-catalyzed carbopalladation and carbocyclization of arynes with aryl halides: a highly efficient route to functionalized triphenylenes. AB - Highly substituted triphenylene derivatives were prepared in good yields via the palladium-catalyzed carbocyclization of arynes with aryl iodides. PMID- 16479302 TI - Fluorous-tagged indolylboron for the diversity-oriented synthesis of biologically attractive bisindole derivatives. AB - The diversity-oriented synthesis of bisindole derivatives to construct concise libraries using consecutive cross-coupling reactions and prepare new sulfonamide type fluorous protecting groups is presented. PMID- 16479304 TI - The "non-nucleophilic" anion [Tf2N]- competes with the nucleophilic Br-: an unexpected trapping in the dediazoniation reaction in ionic liquids. AB - Imidazolium ionic liquids containing [Tf2N]- anion are not as innocent as they are often considered; [Tf2N]- is more reactive than Br- in heterolytic dediazoniation reactions. PMID- 16479305 TI - Mesoporous silicon oxynitride thin films. AB - Highly-ordered, pore-modified with amine groups, and glass-like mesoporous silicon oxynitride thin films were prepared by heat treatment of as-synthesized mesoporous silica thin films in a flowing ammonia environment at high temperatures. PMID- 16479306 TI - Assembly of a novel supramolecular synthon of calix[4]arene presenting four carboxylic acids. AB - An extremely complex solid state structure described by two virtual channels and a 2-D square grid of hydrogen bonds is generated by four carboxylic acids groups of calix[4]arene tetrabutyroxycarboxylic acid. PMID- 16479307 TI - Antibody-functionalized polydiacetylene coatings on nanoporous membranes for microorganism detection. AB - The preparation and characterization of coatings made from polydiacetylene colloids on nano- and microporous membranes and their potential for the detection of microorganisms are presented. PMID- 16479308 TI - A luminescent probe containing a tuftsin targeting vector coupled to a terbium complex. AB - Orthogonal protection strategies have been used to prepare a series of luminescent and MRI active lanthanide complexes containing a tuftsin targeting vector that are internalised by macrophage cells. PMID- 16479309 TI - Superconductivity of doped Ar@C60. AB - The synthesis of a mg amount of pure argon containing fullerene allowed the synthesis of the first endohedral superconductors with critical temperatures lower than expected, an indication of the strong influence of the argon atom on the C60 cage. PMID- 16479310 TI - A modular approach to two-photon absorbing organic nanodots: brilliant dendrimers as an alternative to semiconductor quantum dots? AB - Nanoscopic fluorescent dendrimers having up to 96 two-photon chromophores and showing very large two-photon absorption cross-sections (up to 56 000 GM) were designed as a complementary "organic" alternative to quantum dots. PMID- 16479312 TI - Organic anion transporting polypeptide-C mediates arsenic uptake in HEK-293 cells. AB - Arsenic is an established human carcinogen. The role of aquaglyroporins (AQPs) in arsenic disposition was recently identified. In order to examine whether organic anion transporting polypeptide-C (OATP-C) also plays a role in arsenic transport, OATP-C cDNA was transfected into cells of a human embryonic kidney cell line (HEK 293). Transfection increased uptake of the model OATP-C substrate, estradiol 17beta-D-glucuronide, by 10-fold. In addition, we measured uptake and cytotoxicity of arsenate, arsenite, monomethylarsonate(MMA(V)), and dimethylarsinate (DMA(V)). Transfection of OATP-C increased uptake and cytotoxicity of arsenate and arsenite, but not of MMA(V) or DMA(V). Rifampin and taurocholic acid (a substrate of OATP-C) reversed the increased toxicity of arsenate and arsenite seen in OATP-C-transfected cells. The increase in uptake of inorganic arsenic was not as great as that of estradiol-17beta-D-glucuronide. Our results suggest that OATP-C can transport inorganic arsenic in a (GSH)-dependent manner. However, this may not be the major pathway for arsenic transport. PMID- 16479313 TI - Effects of high dose retinoic acid on TGF-beta2 expression during pancreatic organogenesis. AB - The aim of this study was to investigate the effects of excess all-trans retinoic acid, a vitamin A metabolite, on pancreatic organogenesis and TGF-beta2 expression during prenatal development in rats. First group of animals used as control while a single dose of 60 mg/kg all-trans retinoic acid was ingested by the mothers, at day 8 of gestation (before the neurulation period) in group II and at day 12 of gestation (after the neurulation period) in group III, and all embryos were sacrificed at day 18 of gestation. TGF-beta2 expression was detected in the capsule, acini and Langerhans islets in the control group. In the pancreas of group II, dilatation and congestion of interlobular vessels were observed. Langerhans islet structures were completely absent. Moreover acinar TGF-beta2 immune reactivity was not determined. In group III, acinar expression of TGF beta2 in acid was similar to that in the controls but their Langerhans islets TGF beta2 immune reactivity was significantly less than the controls. In view of the present findings we suggest that TGF-beta2 plays important role in pancreatic morphogenesis and administration of excess all-trans retinoic acid before neurulation inhibit TGF-beta2 expression disrupted pancreatic morphogenesis particularly Langerhans islets. However, its administration after neurulation had less adverse affect on pancreatic organogenesis and TGF-beta2 immune reactivity. PMID- 16479314 TI - Diagnostic value of C-reactive protein in discriminating fungal from nonfungal pulmonary infiltrates in patients with hematologic malignancies. AB - The objective of this study was to assess the utility of C-reactive protein (CRP) on differential diagnosis of pulmonary infiltrates occurring in 143 febrile patients with hematologic malignancies. Serum CRP level, measured on the first day of pneumonia, was significantly higher in patients with fungal lung infiltrates than in those with nonfungal pneumonia (22.3 mg/dl vs 7.3 mg/dl; p<0.0001). Predictive factors for fungal pneumonia were CRP level higher than 10 mg/dl, neutropenia longer than 10 days, and active underlying disease. PMID- 16479316 TI - Fifty years of genetic epidemiology, with special reference to Japan. AB - Genetic epidemiology deals with etiology, distribution, and control of disease in groups of relatives and with inherited causes of disease in populations. It took its first steps before its recognition as a discipline, and did not reach its present scope until the Human Genome Project succeeded. The intimate relationship between genetics and epidemiology was discussed by Neel and Schull (1954), just a year after Watson and Crick reported the DNA double helix, and 2 years before human cytogenetics and the Japan Society of Human Genetics were founded. It is convenient to divide the next half-century into three phases. The first of these (1956-1979) was before DNA polymorphisms were typed, and so the focus was on segregation and linkage of major genes, cytogenetics, population studies, and biochemical genetics. The next phase (1980-2001) progressively identified DNA polymorphisms and their application to complex inheritance. The last phase began with a reliable sequence of the human genome (2002), followed by exploration of genomic diversity. Linkage continues to be useful to study recombination and to map major genes, but association mapping gives much greater resolution and enables studies of complex inheritance. The generation now entering human genetics will have collaborative opportunities undreamed of a few years ago, without the independence that led to great advances during the past half-century. PMID- 16479317 TI - Estimation of haplotype associated with several quantitative phenotypes based on maximization of area under a receiver operating characteristic (ROC) curve. AB - An algorithm for estimating haplotypes associated with several quantitative phenotypes is proposed. The concept of a receiver operating characteristic (ROC) curve was introduced, and a linear combination of the quantitative phenotypic values was considered. This set of values was divided into two parts: values for subjects with and without a particular haplotype. The goodness of its partition was evaluated by the area under the ROC curve (AUC). The AUC value varied from 0 to 1; this value was close to 1 when the partition had high accuracy. Therefore, the strength of association between phenotypes and haplotypes was considered to be proportional to the AUC value. In our algorithm, the parameters representing a degree of association between the haplotypes and phenotypes were estimated so as to maximize the AUC value; further, the haplotype with the maximum AUC value was considered to be the best haplotype associated with the phenotypes. This algorithm was implemented by using R language. The effectiveness of our algorithm was evaluated by applying it to real genotype data of the Calpine-10 gene obtained from diabetics. The results showed that our algorithm was more reasonable and advantageous for use with several quantitative phenotypes than the generalized linear model or the neural network model. PMID- 16479318 TI - The p.T191M mutation of the CBS gene is highly prevalent among homocystinuric patients from Spain, Portugal and South America. AB - Classical homocystinuria is due to cystathionine beta-synthase (CBS) deficiency. More than 130 mutations, which differ in prevalence and severity, have been described at the CBS gene. Mutation p.I278T is very prevalent, has been found in all European countries where it has been looked for with the exception of the Iberian peninsula, and is known to respond to vitamin B6. On the other hand, mutation p.T191M is prevalent in Spain and Portugal and does not respond to B6. We analysed 30 pedigrees from Spain, Portugal, Colombia and Argentina, segregating for homocystinuria. The p.T191M mutation was detected in patients from all four countries and was particularly prevalent in Colombia. The number of p.T191M alleles described in this study, together with those previously published, is 71. The prevalence of p.T191M among CBS mutant alleles in the different countries was: 0.75 in Colombia, 0.52 in Spain, 0.33 in Portugal, 0.25 in Venezuela, 0.20 in Argentina and 0.14 in Brazil. Haplotype analyses suggested a double origin for this mutation. No genotype-phenotype correlation other than the B6-nonresponsiveness could be established for the p.T191M mutation. Additionally, three new mutations, p.M173V, p.I429del and c.69_70+8del10, were found. The p.M173V was associated with a mild, B6-responsive, phenotype. PMID- 16479319 TI - The effect of zinc supplementation on ghrelin-immunoreactive cells and lipid parameters in gastrointestinal tissue of streptozotocin-induced female diabetic rats. AB - Zinc is an essential nutrient with a wide range of functions and closely involved in a variety of enzymatic processes of importance in glucose, protein and lipid metabolism. Ghrelin is the endogenous ligand of the G protein coupled growth hormone secretagogue receptor. The regulatory mechanism that explain the biosynthesis and secretion of ghrelin in the gastrointestinal tract has not been clarified. This study was undertaken to examine the effect of zinc supplementation on the streptozotocin (STZ)-induced diabetic rats, which exhibits ghrelin production and secretion, and lipid metabolism on the gastrointestinal tract. The animals were divided into four groups. Group I: Non-diabetic untreated animals. Group II: Zinc-treated non-diabetic rats. Group III: STZ-induced diabetic untreated animals. Group IV: Zinc-treated diabetic animals. Zinc sulfate was given to some of the experimental animals by gavage at a dose of 100 mg/kg body weight every day for 60 days. In the zinc-treated diabetic group, the blood glucose levels decreased and body weight increased as compared to the diabetic untreated group. Zinc supplementation to STZ-diabetic rats revealed the protective effect of zinc on lipids parameters such as total lipid, cholesterol, HDL-cholesterol and atherogenic index. There is no statistically change in ghrelin-immunoreactive cells in gastrointestinal tissue. But, it has found that zinc supplementation caused a significant reduction in densities of ghrelin producing cells of fundic mucosa of zinc-treated diabetic animals as compared to untreated, non-diabetic controls. Zinc supplementation may contribute to prevent some complications of diabetic rats, biochemically. PMID- 16479320 TI - All-trans retinoic acid induces free radical generation and modulate antioxidant enzyme activities in rat sertoli cells. AB - In this work we investigated the effects of retinoic acid (RA) in Sertoli cells. Sertoli cells isolated from 15-day-old Wistar rats were previously cultured for 48 h and then treated with RA for 24 h. RA at high doses (1-10 microM) increased TBARS levels and induced a decrease in cell viability. At low doses (0.1-100 nM) RA did not increase TBARS level. RA also did not increase cell death at these doses. In order to investigate changes in antioxidant defenses we measured the CAT, SOD and GPx activities in Sertoli cells treated with RA. Compared to control, RA increased around 200% SOD activity in all doses tested (0.1-100 nM); GPx activity was increased 407.49, 208.98 and 243.88% (0.1, 1 and 10 nM, respectively); CAT activity was increased 127% with RA 1 nM. To clarify if RA induces ROS production per se, we performed experiments in vitro using 2 deoxyribose as specific substrate of oxidative degradation by *OH radical as well as TRAP assay. RA at 10 microM increased 2-deoxyribose degradation, suggesting that some of the RA-induced effects are mediated via *OH formation. Furthermore, the total reactive antioxidant potential (TRAP) of the RA was determined. At low concentrations RA has induced no redox activity. Conversely, higher concentration of RA (1-10 microM) increased chemiluminescence. The chemiluminescence produced was directly proportional to radical generation. We provide, for the first time, evidence for a free radical generation by RA. Our results demonstrated that RA plays an important role in Sertoli cells and these effects appear to be mediated by ROS. PMID- 16479321 TI - Effect of ascorbic acid on prevention of hypercholesterolemia induced atherosclerosis. AB - The notion that oxidation of lipids and propagation of free radicals may contribute to the pathogenesis of atherosclerosis is supported by a large body of evidence. To circumvent the damage caused by oxygen free radicals, antioxidants are needed which provide the much needed neutralization of free radical by allowing the pairing of electrons. In this study we have investigated the effect of ascorbic acid, a water soluble antioxidant on the development of hypercholesterolemia induced atherosclerosis in rabbits. Rabbits were made hypercholesterolemic and atherosclerotic by feeding 100 mg cholesterol/day. Different doses of ascorbic acid were administered to these rabbits. Low dose of ascorbic acid (0.5 mg/100 g body weight/day) did not have any significant effect on the percent of total area covered by atherosclerotic plaque. However, ascorbic acid when fed at a higher dose (15 mg/100 g body weight/day) was highly effective in reducing the atherogenecity. With this dose the percent of total surface area covered by atherosclerotic plaque was significantly less (p < 0.001). This suggests that use of ascorbic acid may have great promise in the prevention of hypercholesterolemia induced atherosclerosis. PMID- 16479322 TI - Tiapride pre-treatment in acute exposure to paraoxon: comparison of effects of administration at different points-in-time in rats. AB - INTRODUCTION: Accidental and suicidal exposures to organophosphorus compounds (OPC) are frequent. The inhibition of esterases by OPC leads to an endogenous ACh poisoning. Recently, the FDA approved, based on animal experiments, for military combat medical use oral pyridostigmine (PSTG) for pre-exposure treatment of soman; the concept is to block the cholinesterase reversibly using the carbamate pyridostigmine in order to deny access to the active site of the enzyme to the irreversible inhibitor (OPC) on subsequent exposure. We have shown previously that tiapride (TIA) is in vitro a weak inhibitor of AChE. We also have shown recently that in rats coadministration of TIA with the organophosphate paraoxon significantly decreases mortality without having an impact on red blood cell cholinesterase (RBC-AChE) activity. PURPOSE OF THE STUDY: To establish in a prospective, non-blinded study in a rat model of acute high dose OPC (paraoxon; POX) exposure the ideal point in time for TIA pre-treatment administration and to correlate it with measured TIA plasma levels. MATERIAL AND METHODS: There were six groups of rats in each cycle of the experiment and each group contained six rats. The procedure was repeated twelve times (cycles) (n = 72 for each arm; half male and half female). All substances were applied ip. All groups (1-6) received 1 microMol POX ( approximately LD(75)); groups 1-5 also received 50 microMol TIA at different points in time. Group 1 (G(1)): TIA 120 min before POX Group 2 (G(2)): TIA 90 min before POX, Group 3 (G(3)): TIA 60 min before POX, Group 4 (G(4)): TIA 30 min before POX, Group 5 (G(5)): TIA & POX simultaneously, Group 6 (G(6)): POX only. The animals were monitored for 48 hours and mortality/survival times were recorded at 30 min, 1, 2, 3, 4, 24 and 48 h. AChE activities were determined at 30 min, 24 and 48 h in surviving animals. Statistical analysis was performed on the mortality data, cumulative survival times and enzyme activity data. Mortality data was compared using Kaplan-Meier plots. Cumulative survival times and enzyme activites were compared using the Mann-Whitney rank order test. No Bonferroni correction for multiple comparisons was applied and an alpha < or= 0.05 was considered significant. RESULTS: Mortality is statistically significantly reduced by TIA pre-treatment at all points-in-time. Highest protection is achieved if TIA is given 90 to 0 min before OPC exposure. The reduction in mortality is not correlated to TIA plasma levels (C (max) approximately 120 min post ip-administration). TIA pre-treatment is not affecting AChE activity regardless of the timing of administration. CONCLUSION: The lack of correlation between TIA plasma levels and degree of mortality reduction as well as the lack of protective effect on enzyme activity seem to indicate that the site of action of TIA is not the blood. While our hypothesis that TIA would protect AChE in a pyridostigmine-like manner (via protection of the enzyme) could not be confirmed, the reduction in mortality with TIA pre-treatment is nevertheless of potential interest. PMID- 16479323 TI - Design and analysis of association studies using pooled DNA from large twin samples. AB - Evidence is mounting that multiple genes are involved in complex traits and that these each account for very small proportions of the overall phenotypic variance. Association studies of many markers in 1000s of individuals will be required to identify such genes. A number of large twin cohorts have already been collected and provide a valuable resource for carrying out studies that are robust to the effect of population stratification. Technologies based on microarrays will soon allow 1.000,000 SNPs to be typed at one time, however financial considerations prevent most researchers from using these approaches to genotype all individuals. Recently, microarrays have been shown to give accurate allele frequency measurements in pooled DNA samples and provide a simple way to select the best markers for individual genotyping. This drastically reduces the cost and workload of large scale association studies. One limitation of this methodology relates to the analytical procedures which have only been developed to allow comparison of two pools e.g. case/control pools. In this paper we use meta-regression to analyze pooled DNA data allowing the allele frequency in each pool to be related to the average quantitative phenotypic measure of the individuals whose DNA were used to construct the pools. Alongside this we describe a technique that can be used to determine the power for such studies. We present results from some preliminary investigations of different pooling strategies that can be applied to large twin samples and demonstrate that the method retains a large proportion of the power available from individual genotyping. PMID- 16479324 TI - Lifetime psychiatric comorbidity with substance use disorders: does healthcare use modify the strength of associations ? AB - BACKGROUND: The frequent psychiatric comorbidity among subjects with a substance use disorder (SUD) can be explained by an increased vulnerability to problematic drug use among subjects with a non-substance-related psychiatric disorder (NSRPD). The care of this disorder should then reduce the likelihood of a secondary SUD. OBJECTIVE: To examine how healthcare use for psychological symptoms modifies the lifetime association between SUD and NSRPD. METHODS: Two hundred and twenty four students were evaluated for mental disorders and healthcare use. Mental disorders were assessed with the Mini-International Neuropsychiatric Interview (MINI). Healthcare use included consultations with a general practitioner (GP), a psychiatrist or a psychologist. The lifetime occurrence of a SUD was analysed by lifetime number of NSRPD and healthcare use for psychological symptoms. Analyses were adjusted on gender, university affiliation, living environment and temperament and character dimensions. RESULTS: Compared to subjects without NSRPD, those with at least two NSRPD who did not use healthcare were more likely to have had a lifetime SUD (OR = 3.9). By contrast, those who had only one NSRPD seemed to be as likely as those with no NSRPD to have had a SUD. DISCUSSION: These results suggest a decreased probability of SUD among subjects with several NSRPD who used healthcare. Due to the cross-sectional design of this study, causal inferences cannot be drawn. This analysis shows the importance, however, of taking healthcare use into account in comorbidity studies. PMID- 16479325 TI - Stressors and psychological symptoms in students of medicine and allied health professions in Nigeria. AB - BACKGROUND: Studies suggest that high levels of stress and psychological morbidity occur in health care profession students. This study investigates stressors and psychological morbidity in students of medicine, dentistry, physiotherapy and nursing at the University of Ibadan. METHODS: The students completed a questionnaire about their socio-demographic characteristics, perceived stressors and the 12-item General Health Questionnaire. Qualitative methods were used initially to categorise stressors. Data was then analysed using univariate and logistic regression to determine odds ratios and 95% confidence intervals. RESULTS: Medical and dental students were more likely to cite as stressors, overcrowding, strikes, excessive school work and lack of holidays while physiotherapy and nursing students focused on noisy environments, security and transportation. Medical and dental students (1.66; SD: 2.22) had significantly higher GHQ scores than the physiotherapy and nursing students (1.22; SD: 1.87) (t = 2.3; P = 0.022). Socio-demographic factors associated with psychological morbidity after logistic regression include being in a transition year of study, reporting financial distress and not being a 'Pentecostal Christian'. Although males were more likely to perceive financial and lecturer problems as stressors and females to perceive faculty strikes and overcrowding as source of stress, gender did not have any significant effect on psychological morbidity. Stressors associated with psychological distress in the students include excessive school work, congested classrooms, strikes by faculty, lack of laboratory equipment, family problems, insecurity, financial and health problems. CONCLUSION: Several identified stressors such as financial problems, academic pressures and their consequent effect on social life have an adverse effect on the mental health of students in this environment especially for students of medicine and dentistry. While stressors outside the reach of the school authorities are difficult to control, academic support including providing a conducive learning environment, advice on means for sustenance, added support during periods of transition are key areas for interventions. PMID- 16479326 TI - Factors associated with receiving help and risk factors for disaster-related distress among Connecticut adults 5-15 months after the September 11th terrorist incidents. AB - BACKGROUND: To obtain prevalence estimates and identify factors associated with psychological problems and receipt of help by a geographically proximate population in which some persons had direct exposure but the overall prevalence of direct exposure was low, 5-15 months after the September 11th, 2001 terrorist incidents. METHOD: Telephone survey data from the Behavioral Risk Factor Surveillance System (BRFSS) (Connecticut Module) with a randomly selected cohort of 2741 women and 1899 men aged 18 and older were examined using bivariate Chi Square and multivariate logistic regression analyses of weighted data. RESULTS: One in three respondents reported 9/11-related psychological problems, 26% of whom reported receiving formal services or peer support. Risk factors for reporting psychological problems included being surveyed earlier, female gender, age 64 or younger, Hispanic ethnicity, disability, recent depression, and reporting one day or more in the past 30 of poor mental health, sleep problems or worry. Poor mental health was associated with receipt of formal services, and increased alcohol use was associated with receipt of peer support. CONCLUSION: In the post-impact recovery period following mass trauma, psychological problems by persons with ongoing mental health conditions or increased alcohol use warrant continuing public and professional attention. Women, Hispanics, and disabled adults also may be under-served. PMID- 16479327 TI - Need and utilization of psychiatric consultation services among general hospital inpatients. AB - OBJECTIVE: The aim of the present study was to estimate the need for psychiatric consultation services and psychiatric inpatient referral among inpatients of non psychiatric hospital departments, and to assess the actual utilization of these services. METHODS: The study was carried out among 728 inpatients of physical rehabilitation wards, medical, surgical, and gynecological wards in Austrian hospitals. Psychiatric case identification was performed by research psychiatrists using the Clinical Interview Schedule (CIS). Diagnoses were given according to DSM-III-R. The assessment of need for consultation and inpatient referral was based on the clinical judgments of research psychiatrists. RESULTS: Overall, 34.2% of the inpatients were cases according to the CIS-criteria, 51.8% of them needing either psychiatric consultation or inpatient referral according to research psychiatrists. In 66.7% of those for whom research psychiatrists had stated a need, this need was not met ("unmet need"), while only 33.3% of them had their need met. In contrast, a psychiatric consultation was performed among 5% of those patients not needing psychiatric services according to the research psychiatrists ("overprovision"). Variables of the health care system (i.e. department type and catchment area of the hospital) were among the predictors for "met needs". CONCLUSION: The rate of actual psychiatric consultations and admissions to psychiatric wards was markedly lower than the need according to research psychiatrists' judgment. PMID- 16479329 TI - Vocal fold paralysis as a sign of chest diseases: a 15-year retrospective study. AB - BACKGROUND: Vocal fold paralysis (VFP) is sometimes the only sign of chest diseases. However, some patients with VFP due to chest diseases are not diagnosed correctly at the first examination, which may leave the patients untreated for a long time. Depending on the situation, chest x-ray is not enough for detecting the primary lesion. The objective of this study was to discuss the diagnostic procedure for VFP based on the retrospective analysis of the cases. METHODS: A total of 42 patients (29 males and 13 females) with VFP due to chest disease examined at the Department of Otolaryngology of Kyoto Prefectural University of Medicine between 1988 and 2002 were reviewed retrospectively. RESULTS: Of the primary chest diseases, lung cancer (15 cases) was the most common, followed by thoracic aortic aneurysm (TAA) (9 cases), metastatic tumor from other regions (6 cases), pulmonary and mediastinal tuberculosis (TB) (5 cases), and esophageal cancer (4 cases). While the primary lesions were easily detected with chest x-ray in most of the cases, some lesions in the aortopulmonary window were difficult to detect. Contrast-enhanced computed tomography (CT) was useful to detect any mass in this region. CONCLUSIONS: In the diagnosis of VFP due to chest diseases, chest x-ray was useful but not always enough for detecting the primary lesion. Necessity of further examinations including contrast-enhanced chest CT must be kept in mind for the cases with negative chest radiographs. PMID- 16479328 TI - Popliteal venous aneurysm--a systematic review. AB - BACKGROUND: Popliteal vein aneurysm is a rare but potentially problematic disease with a risk of pulmonary embolism. METHOD: A systematic literature search was performed. RESULTS: A total of 105 popliteal vein aneurysms were identified, with a slight female and left-sided preponderance. Dominating symptoms are caused by pulmonary embolism, followed by local symptoms with a palpable mass. In four patients arterial symptoms were reported. The most frequently used diagnostic test was phlebography, increasingly replaced by duplex ultrasonography in recent years. Because pulmonary embolism is a potential, it motivates surgical removal. In most cases excision and lateral suture is possible. Four fatal cases were reported. Unfortunately follow-up time and results are poorly documented. CONCLUSION: Popliteal venous aneurysm is rare but should be considered as a local source of pulmonary embolism when no other explanation is found. PMID- 16479330 TI - Prognostic factors of primary small bowel adenocarcinoma: univariate and multivariate analysis. AB - BACKGROUND: Adenocarcinoma of the small bowel is relatively less common than malignancies of the esophagus, stomach, and colorectum. In small bowel adenocarcinoma, various prognostic factors influence the disease-free status and overall survival rates. MATERIALS AND METHODS: Eighty patients who were diagnosed with small bowel adenocarcinoma and treated at our institute between 1983 and 2003 were retrospectively reviewed. RESULTS: The patients included 40 men and 40 women with an age range of 15 to 93 years (median: 62 years). Only 51.3% of patients were accurately proved preoperatively to have a malignancy by endoscopic biopsy. Sixty patients underwent surgical treatment, and 45 of those patients had curative resection. The follow-up period ranged from 2.5 to 229.7 months, with a median of 9.1 months. The cumulative 1-, 3-, and 5-year survival rates for all patients (excluding 3 patients who died in the immediate postoperative period) were 43.6%, 22.8%, and 17.5%, respectively. The cumulative 1-, 3-, and 5-year disease-free survival rate for all 43 patients with curative resection (excluding 2 patients who died in the immediate postoperative period) was 54.9%, 30.5%, and 27.4%, respectively. Meanwhile, multivariate analysis with Cox proportional hazards analysis demonstrated that earlier tumor stages (stages I and II) and curative resection were two independent factors influencing favorable overall survival. Lymph node metastasis was the only independent factor predicting poor disease-free survival in patients undergoing curative resection. CONCLUSIONS: Poor prognosis of small bowel adenocarcinoma may be related to a delay in the diagnosis and treatment of the disease. Curative resection is the aim of surgical treatment for small bowel adenocarcinoma. Lymph node metastasis at presentation of the disease predicts tumor recurrence and distant metastasis after curative surgical treatment. PMID- 16479331 TI - Factors associated with disease survival after surgical resection in Chinese patients with hepatocellular carcinoma. AB - OBJECTIVE: The aim of this cohort study was to investigate clinical outcome and prognostic factors after surgical resection for hepatocellular carcinoma (HCC). MATERIALS AND METHODS: A total of 1,157 HCC patients undergoing hepatic resection between 1998 and 2003 were included in this study. Univariate and multivariate analyses were performed to examine factors affecting clinical outcome and recurrence. RESULTS: Surgical procedures consisted of 1,011 (87.4%) anatomical resections, including 205 (17.7%) extended hepatectomies, 324 (28.0%) hemihepatectomies, 482 (41.7%) segmental resections, and 146 (12.6%) local resections. The results suggest that 56.6% of patients had a recurrence of HCC during the study period and the main recurrence type was intrahepatic (542; 83.1%). The median survival time was 45 months. The 1-, 3- and 5-year overall disease-free survival rates for the study population were 74%, 47%, and 39% respectively. CONCLUSIONS: The results of proportional hazard analyses suggest that tumor size, number of nodules and vascular invasion were significant predictors for poor survival rates. PMID- 16479332 TI - To the editor: Is von Willebrand factor a hormone that regulates a coagulation mechanism? PMID- 16479333 TI - Dilemma of general surgery recruitment in rural America. PMID- 16479334 TI - Prospective comparison of laparoscopic left hemicolectomy for colon cancer with laparoscopic left hemicolectomy for benign colorectal disease. AB - BACKGROUND: Short-term outcome and anorectal function results after laparoscopic hemicolectomy for colon cancer were compared with results after laparoscopic hemicolectomy for benign diseases. METHODS: A total of 108 patients who underwent laparoscopic left colectomy (60 for colon cancer, 48 for diverticulitis or polyposis) were enrolled in the study. Left hemicolectomy in patients affected by cancer was performed by high ligation of the inferior mesenteric artery. A questionnaire concerning anorectal function was mailed to patients 6 months after surgery. RESULTS: Complications were more frequent in the cancer group than in the benign disease group: overall morbidity rate (29.6% versus 8.7%; P = 0.009), diarrhea during the first 6 postoperative months (58.7% versus 34.1%; P = 0.022), and anorectal function problems (fecal incontinence and/or the inability to discriminate between gas and stool, and/or urgency, and/or tenesmus) (65.2% versus 31.7%; P = 0.002). DISCUSSION: The level of ligation of the lower mesenteric artery and damage at the lower mesenteric ganglion could explain the poorer anorectal function outcome in the colon cancer group. PMID- 16479337 TI - Perforation of the gastrointestinal tract secondary to ingestion of foreign bodies. AB - INTRODUCTION: Ingesting a foreign body (FB) is not an uncommon occurrence. Most pass through the gastrointestinal (GI) tract uneventfully, and perforation is rare. The aim of this study was to report our experience with ingested FB perforations of the GI tract treated surgically at our institution. METHODS: A total of 62 consecutive patients who underwent surgery for an ingested FB perforation of the GI tract between 1990 and 2005 were retrospectively reviewed. Three patients with no definite FB demonstrated intraoperatively were included. RESULTS: The patients had a median age of 58 years, and 37 (60%) were male. Of the 59 FBs recovered, 55 (93%) were toothpicks and dietary FBs such as fish bones or bone fragments. A definitive preoperative history of FB ingestion was obtained for only two patients, and 36 of 52 patients (69%) wore dentures. Altogether, 18 (29%) perforations occurred in the anus or distal rectum, and 44 perforations were intraabdominal, with the most common abdominal site being the distal ileum (39%). Patients with FB perforations in the stomach, duodenum, and large intestine were significantly more likely to be afebrile (P = 0.043), to have chronic symptoms (> 3 days) (P < 0.001), to have a normal total white blood cell count (P < 0.001), and to be asymptomatic or present with an abdominal mass or abscess (P < 0.001) compared to those with FB perforations in the jejunum and ileum. CONCLUSIONS: Ingested FB perforation in the adult population is most commonly secondary to unconscious accidental ingestion and is frequently caused by dietary FBs especially fish bones. A preoperative history of FB ingestion is thus rarely obtained, although wearing dentures is a common risk factor. FB perforations of the stomach, duodenum, and large intestine tend to present with a longer, more innocuous clinical picture than perforations in the jejunum or ileum. PMID- 16479338 TI - The rural surgeon: an endangered species. PMID- 16479336 TI - Effect of temporary abdominal closure on colonic anastomosis and postoperative adhesions in experimental secondary peritonitis. AB - BACKGROUND: The effect of relaparotomies and temporary abdominal closure on colonic anastomoses and postoperative adhesions is under debate. METHODS: In the experiments reported here, colonic anastomosis was constructed 24 hours after cecal ligation and puncture in rats that were divided into three groups of eight animals each. The abdomen was closed primarily in groups I and II, and a Bogota bag was used for abdominal closure in group III. At 24 hours following anastomosis, relaparotomy was performed only in group II and III rats, and the abdomen was closed directly in group II; after removal of the Bogota bag in group III animals, the abdomen was closed directly. On the fifth day of anastomotic construction, bursting pressures and tissue hydroxyproline content of the anastomoses, along with peritoneal adhesions, were assessed and compared. RESULTS: Mean anastomotic bursting pressures and hydroxyproline contents did not differ among the groups. Median adhesion scores were significantly higher in group III than the other two groups. CONCLUSIONS: Relaparotomy and the type of temporary closure have no negative effect on anastomotic healing in rats with peritonitis. Temporary abdominal closure with a Bogota bag caused a significantly high rate of adhesions. PMID- 16479339 TI - Comparison of Kugel and Lichtenstein operations for inguinal hernia repair: results of a prospective randomized study. AB - INTRODUCTION: The aim of this prospective randomized clinical study was to compare the Lichtenstein hernia repair with Kugel's patch hernia repair. METHODS: From September 1999 to August 2002, adult patients with inguinal hernia were randomized into two groups. Group I included patients with the Lichtenstein hernioplasty, and group II included patients with the Kugel hernioplasty. The duration of the operation, surgical findings, and postoperative complications were recorded for the patients in both groups. RESULTS: A total of 139 patients (134 men, 5 women) were randomized to either group I or group II. No significant differences were observed for the duration of the operation or the complication rates between the groups during the same follow-up time (53.06 +/- 5.6 months vs. 53.41 +/- 7.11 months in groups I and II, respectively). CONCLUSIONS: Kugel herniorraphy is a minimally invasive method and as safe as the Lichtenstein hernia repair technique, with similar complication rates. PMID- 16479340 TI - The efficacy of intraoperative corticosteroids in recurrent laryngeal nerve palsy after thyroid surgery. AB - BACKGROUND: Recurrent laryngeal nerve palsy (RLNP) is one of the most common complications after thyroidectomy. We aim to evaluate the role of intraoperative corticosteroids in preventing or treating postoperative RLNP. MATERIALS AND METHODS: We conducted a prospective study of 295 patients who underwent either total lobectomy or total thyroidectomy with or without using intraoperative corticosteroids. All cases were operated on by the same surgeon. Measurement of the RLNP rate was based on the number of nerves at risk. Data were analyzed for differences in postoperative temporary or permanent RLNP rate and recovery days. RESULT: The rate of temporary/permanent RLNP was 5.7% (11 out of 194)/0.52% (1 out of 194) and 6.9% (12 out of 173)/0.58% (1 out of 173) according to groups with and without corticosteroids. This difference did not reach statistical significance. Among those 23 patients who recovered from RLNP, the mean time to recovery was significantly shorter for patients receiving intraoperative steroids (28.6 vs. 40.5 days, P = 0.045). CONCLUSION: A single dose of intraoperative corticosteroids did not produce any benefit in terms of reducing the postoperative temporary/permanent RLNP rate, but it did shorten the recovery time for patients suffering from temporary RLNP. PMID- 16479341 TI - Does familial non-medullary thyroid cancer adversely affect survival? AB - BACKGROUND: Familial non-medullary thyroid cancer (FNMTC) is associated with a higher rate of multifocality and a higher recurrence rate than sporadic thyroid cancer. However, the effect of FNMTC on life expectancy is unknown. MATERIAL AND METHODS: Using data from our FNMTC database, we calculated life expectancy and survival rates after diagnosis of FNMTC and compared the results with the rates for unaffected family members and for the standard US population. Overall life expectancy and survival rates were calculated using the Kaplan-Meier method. We compared patients from families with 2 affected members with patients from families with > or = 3 affected members. We also compared patients diagnosed in a known familial setting (index cases and subsequent cases) with patients diagnosed before the familial setting was recognized. RESULTS: There were 139 affected patients with 757 unaffected family members. The mean age at diagnosis was 40.8 +/- 13.9 years and the mean follow-up time was 9.4 +/- 11.7 years. Ten patients died of thyroid cancer during follow-up. The life expectancy of patients with FNMTC was similar to that of their unaffected family members. Survival was significantly shorter for patients with 3 or more affected family members, for patients diagnosed before the familial setting was recognized, and for patients with anaplastic cancer. CONCLUSIONS: Our results suggest that FNMTC may be more aggressive than sporadic thyroid cancer, particularly in families with 3 or more affected members. However, when recognized and treated appropriately, it does not significantly shorten the overall life expectancy of the affected patients. PMID- 16479342 TI - A prophylactic infusion of calcium solution reduces the risk of symptomatic hypocalcemia in patients after total thyroidectomy. AB - BACKGROUND: Several risk factors have been associated with post-operative transient hypocalcemia after thyroid surgery. However, there are no studies evaluating preventive measures to avoid symptomatic postoperative hypocalcemia. Although intravenous infusion of calcium improves hypocalcemic symptoms, it is unknown whether prophylactic infusion prevents symptoms of postoperative hypocalcemia. PATIENTS AND METHODS: Five hundred and forty-seven patients underwent total thyroidectomy. Two groups were identified: group A (n = 243) received prophylactic intravenous drip infusion of 78-156 mg of calcium solution at 3-8 hours after operation, and group B (n = 304) received no prophylactic treatment. Prophylactic infusion was used only once if the patients did not have symptoms of hypocalcemia. Serum calcium (Ca) levels, intact parathyroid hormone (i-PTH) levels on the first postoperative day (1st POD), and the prevalence of symptoms of hypocalcemia were prospectively analyzed. RESULTS: The serum Ca levels at the 1st POD in group A patients (7.91 +/- 0.49 mg/dl, mean +/- SD) was significantly higher than group B patients (7.65 +/- 0.54, P < 0.0001), while the serum i-PTH levels were not significantly different between the two groups. The prevalence of numbness and/or tetany before noon on the 1st POD was significantly lower in group A patients. Prophylactic infusion of calcium solution reduced the prevalence of tetany from 8.6% to 2.1%. CONCLUSION: A prophylactic infusion of calcium solution after total thyroidectomy may be useful in reducing the development of symptomatic hypocalcemia and reduces the patients' risk of having discomfort and anxiety due to hypocalcemia. PMID- 16479343 TI - Carbon dye staining of sentinel lymph nodes facilitates microstaging of colon cancer patients. AB - BACKGROUND: Carbon dye, when peritumourally injected, permanently marks the drainage site of sentinel lymph nodes (SLN). The objective of the current study was to evaluate whether the use of carbon dye facilitated the detection of small nodal tumour infiltrates in colon cancer patients. METHODS: In a prospective trial, 19 patients underwent open, oncological resections of localized colon cancer and SLN procedure according to a standardized protocol. Isosulfan blue 1% and sterile filtered carbon dye (mixed 1:1) were injected into the subserosa circumferentially around the tumour. Lymph nodes staining blue were marked as SLN. Serial sections of each SLN were stained with hematoxylin and eosin (H&E) and with the pancytokeratin marker AE1/AE3. The intranodal presence and site of carbon particles were noted and compared with the location of possible tumour infiltrates. RESULTS: Identification of at least one SLN was successful in 18 patients (identification rate 95%). Four patients (22%) were pN+, 11 (61%) were pN0(i-). Three patients (17%) were upstaged from pN0(i-) to pN0(i+) as isolated tumour cells were detected in their SLN: in two (11%) of the three patients, carbon dye and isolated tumour cells were found in the same nodal compartment, hence facilitating the recognition of isolated tumour cells by the pathologist. CONCLUSION: The use of carbon dye in the SLN procedure for colon cancer may facilitate the detection of small nodal tumour infiltrates. PMID- 16479344 TI - Training in laparoscopic appendectomy. AB - BACKGROUND: The role of laparoscopic appendectomy (LA) in surgical training is unclear. Although LA as a therapeutic modality is potentially superior to open surgery, it has failed to become established as standard in training hospitals. The aim of the present study was to evaluate the outcome of LA performed by inexperienced surgeons in a training environment. MATERIALS AND METHODS: A retrospective analysis of all attempted LA performed over a 12-month period was undertaken. Data collected included operator grade (experienced and inexperienced), conversion rate and duration of surgery, complications, and postoperative stay. RESULTS: During the study period, 169 appendectomies were performed. The conversion rate to open surgery declined significantly from 28% in the first quarter to 9% in the last quarter, with no difference in the conversion rate between experienced and inexperienced surgeons. Operative time shortened significantly in the inexperienced group. Postoperative complications occurred in 8% of patients, independent of operative grade. CONCLUSIONS: Our findings demonstrate that LA may be safely introduced as a teaching procedure. Time-to train should not preclude institutions from adopting the laparoscopic approach in the treatment of acute appendicitis. PMID- 16479345 TI - Incidence of vascular injuries after use of the Angio-Seal closure device following endovascular procedures in a single center. AB - INTRODUCTION: Percutaneous closure devices have been used to obtain rapid hemostasis and early mobilization of the patient after arterial catheterization. However, we observed challenging problems with the sealing procedure that require further surgical intervention. The present report is a retrospective analysis of the patterns of injury and the final outcome of four cases of femoral artery injury following the use of Angio-Seal. METHODS: During the last 24 months, in a group of 175 patients (131 men, 44 women; median age 68.4 years, range 47-81 years) underwent percutaneous closure after diagnostic (n = 53) or therapeutic (n = 122) endovascular procedures. Among them we observed four patients (three men, one woman; median age 65.2 +/- 10.8 years, range 47-75 years) who developed severe limiting claudication and required vascular repair of an iatrogenic vascular injury following deployment of the Angio-Seal. They had a femoral thrombosis due to narrowing/severe intimal dissection. RESULTS: All patients required operative intervention with removal of the device. We performed femoropopliteal thrombectomy and common femoral endarterectomy with patch angioplasty (n = 2), resection of the femoral bifurcation and reimplantation of the deep femoral artery (n = 1), and femoral bifurcation endarterectomy with direct arterial suture (n = 1). The median hospital stay was 6.5 +/- 3.8 days (range 4-12 days). Limb salvage was achieved in all of the surviving patients at a mean follow-up of 7 months (range 1-12 months). CONCLUSIONS: Vascular injuries are uncommon after use of the hemostasis closure device. When they occur, however, they are likely to require challenging surgical correction. PMID- 16479346 TI - Randomized controlled trial of LigaSure with submucosal dissection versus Ferguson hemorrhoidectomy for prolapsed hemorrhoids. AB - INTRODUCTION: The aim of this study was to compare the outcomes between the LigaSure vessel sealing system and the conventional closed Ferguson hemorrhoidectomy procedure performed by diathermy. METHODS: A series of 84 patients with grades III and IV hemorrhoids were randomized into two groups: (1) LigaSure hemorrhoidectomy with submucosal dissection (42 patients) and (2) Ferguson hemorrhoidectomy (42 patients). The patient demographics, operative details, parenteral analgesic requirement, postoperative pain score (assessed by an independent assessor), operating time, intraoperative blood loss, hospital stay, early and delayed complications, and time off from work or normal activity were recorded. The patients were regularly followed up at 1, 2, 4, 6, and 8 weeks after surgery. RESULTS: There were no statistically significant differences between the two groups in terms of age, gender, duration of symptoms, grade of the hemorrhoid(s), or number of hemorrhoids resected. The mean operating time for LigaSure hemorrhoidectomy with submucosal dissection was significantly shorter than that for the Ferguson hemorrhoidectomy (11.3 +/- 0.4 vs. 34.2 +/- 0.7 minutes; P < 0.0001). Patients treated with the LigaSure method had significantly less blood loss (P < 0.0001), a better pain score (P < 0.0001), less parenteral analgesic requirement (P < 0.0001), shorter hospital stay (P < 0.0001), and less time off from work or normal activity (P < 0.0001). There was no difference in the early and delayed postoperative complications between the two groups. CONCLUSIONS: LigaSure hemorrhoidectomy with submucosal dissection is a safe, effective procedure for grade III and IV hemorrhoids. Patients derive greater short-term benefits: reduced intraoperative blood loss, operating time, and postoperative pain as well as earlier resumption of work or normal activity. Long term follow-up with a larger number of patients is required to confirm the long term results of this procedure. PMID- 16479347 TI - New prognostic scales LAST-1 and LAST-2: supporting prediction and staging of thyroid cancer. AB - INTRODUCTION: Epidemiologically, thyroid gland tumors are lesions of the highest importance among endocrine tumors in humans. Although the results of surgical treatment of the highly differentiated (follicular and papillary) tumors seem to be satisfactory, treatment of the poorly differentiated (medullary and anaplastic) tumor still demands clinical and basic investigations. In this study the authors sought to evaluate clinical and molecular factors that could contribute to preoperative detection of more advanced thyroid cancers (i.e., those that exhibit extrathyroid spread and lymph node invasion). METHODS: A total of 27 patients operated on for thyroid cancer were evaluated according to age, sex, time from the onset of the disease, cytogenetic changes, and loss of heterozygosity (LOH) in 14 microsatellite markers. The output variables were defined according to postoperative findings and the TNM 2002 score. The T1-2 N0 M0 cases were defined as local malignancy (LM); and T3-4 any N any M, any T N1 any M, or any T any N M1 were considered advanced malignancy (AM). The control groups consisted of 25 patients with multinodular goiter (MNG) and 32 patients with follicular adenoma (FA). In all cases, clinical and molecular data similar to those listed above were collected, excluding staging and follow-up information. RESULTS: There was no predominant specific type of chromosomal aberration observed and no marker lost in more than five patients (18%). The logistic regression identified three input variables as contributing significantly to the dichotomized outcome measure (LM vs. AM): LOH in any of the examined loci, age of the patient at the presentation, and the sex of the patient. Furthermore, discriminant analysis revealed four input variables differentiating among TC, FA, and MNG patients. Based on the multivariate analysis results, two numeric prognostic scales were fashioned: LAST-1, a scale applicable to differentiation of thyroid cancers at different degrees of clinical advancement; and LAST-2, a scale applicable to differentiation of any thyroid lumps. CONCLUSIONS: It was concluded that LOH and the age and sex of the patients can provide sufficient data to predict thyroid cancer with a high degree of clinical advancement. LAST-1 scale is a reliable tool for identifying these patients. The LAST-2 scale gives supportive information about the character of thyroid lumps, distinguishing TC from MNG and FA. PMID- 16479349 TI - Treatment of bladder tumors and benign prostatic hyperplasia with a new TUR system using physiological saline as perfusate. AB - This article presents a new transurethral resection (TUR) system for use in endoscopic surgery. By using an electroconductive solution (physiological saline) as the perfusate in lieu of conventional non-electroconductive solution (Uromatic), additional anesthesia (e.g., obturator nerve blocking) is not required. The new TUR is carried out in an electroconductive solution such as saline, and because radiofrequency current flows from the resecting electrode through the perfusate to the outer sheath, no counter-electrode is needed. We have treated both bladder tumor and benign prostatic hyperplasia cases with this new system. Surgery was safely performed in all TUR-bt cases without requiring obturator nerve blocking. During both TUR-bt and transurethral resection of the prostate (TUR-P) using this system, tissue resection and coagulation equivalents were similar to the conventional TUR system. In previous TUR, preoperative obturator nerve blocking was necessary, and in some cases, incomplete blocking or complications occurred. When physiological saline is used as the perfusate, blood electrolyte levels are not greatly changed, even after extensive resection of the bladder wall; as a result, this new system is also cost effective because physiological saline is less expensive than non-electroconductive solutions and requires no counter-electrode. Thus, in comparison with conventional TUR, this new system is both significantly safer and more cost effective. PMID- 16479348 TI - Tissue oxygen saturation during colorectal surgery measured by near-infrared spectroscopy: pilot study to predict anastomotic complications. AB - INTRODUCTION: We investigated the relation between tissue oxygen saturation measured by near-infrared spectroscopy (NIRS) and anastomotic complications associated with colorectal surgery. METHODS: A series of 20 patients with colorectal cancer underwent radical surgery with enteric anastomosis. Measurements of tissue oxygen saturation (StO2) were performed at both the proximal and distal portions of the anastomotic site; in cases of anterior resection, we measured StO2 only in the proximal portion. RESULTS: Two anastomotic complications (one leakage, one stenosis) occurred in the 20 cases. The StO2 in patients with anastomotic complications was 58.0%, and that in patients without complications was 71.0%. Altogether, 18 patients had StO2 values > 66%, and none of them had anastomotic complications. In contrast, 2 patients had StO2 values < 60%, and both had anastomotic complications. CONCLUSIONS: The StO2 of the anastomotic site can be safely and reliably measured by NIRS during colorectal surgery. Low StO2 on both sides of the anastomosis may indicate an increased risk of anastomotic complications. Further study is needed to determine the cutoff value for StO2 required to prevent serious complications. PMID- 16479351 TI - Video-assisted thoracoscopic surgery for recurrent spontaneous pneumothorax: the long-term benefit. AB - BACKGROUND: Management of recurrent primary spontaneous pneumothorax by open surgery was considered the treatment of choice until recently. The major drawbacks of this management are the prolonged postoperative pain and cosmetic results. In the last decade, video-assisted thoracoscopic surgery (VATS) has replaced the routine use of open surgery. Most papers that compared limited open surgery to VATS addressed the early postoperative results, and studies that assessed the long-term results focused primarily on the rate of recurrence and pulmonary function tests. The aim of this study was to compare the outcome of minithoracotomy and VATS with emphasis on patients' long-term, subjective perspective and satisfaction. METHODS: Medical records of patients with recurrent primary spontaneous pneumothorax were retrospectively reviewed. Patients who underwent surgical treatment by limited thoracotomy (63 patients) or VATS (58 patients) more than 3 years ago were enrolled. Hospital medical charts were used to compare the early postoperative results. Outpatient clinic records and a telephone questionnaire were employed to evaluate long-term results. RESULTS: There was no mortality or major morbidity in either group, and hospitalization time was similar. Patients in the thoracotomy group needed significantly higher doses of narcotic analgesia for a longer period. There were two cases of recurrence in the VATS group (3%). Seventy-eight percent of patients in the VATS and 21% in the thoracotomy group classified their pain as insignificant a month following the operation (P < 0.05). Three years following surgery, 97% of the VATS group patients considered themselves completely recovered from the operation compared with only 79% in the thoracotomy group (P < 0.05). Nineteen percent of the thoracotomy group and 3% of the VATS group suffered from chronic or intermittent pain necessitating use of analgesics more than once a month. Thirteen percent of patients from the open procedure group required services from the pain clinic. Patients in the VATS group were, in general, much more satisfied with their operation and with the surgical scars compared with patients from the thoracotomy group. CONCLUSION: We recommend video-assisted surgery as the first line surgical treatment for patients with recurrent primary spontaneous pneumothorax. This recommendation is based on its somewhat favorable early postoperative course, the superior long-term outcome, and patient satisfaction. PMID- 16479350 TI - New indication for reduction surgery in patients with advanced hepatocellular carcinoma with major vascular involvement. AB - BACKGROUND: The prognosis of advanced hepatocellular carcinoma (HCC) remains poor, particularly in patients with tumor thrombi (TT) in the major vessels. PATIENTS AND METHODS: From July 1992 to October 2004, 161 patients diagnosed as having advanced HCC with major vascular involvement were seen consecutively at our hospital. Among these patients, 32 (20%) underwent surgical resection [16 complete resection (CR), 16 reductive resection (RR)]. Eighteen patients (11%) received radiotherapy (RT), 73 (45%) underwent transcatheter arterial chemoembolization (TACE) or transcatheter arterial infusion chemotherapy (TAI), 8 (5%) with distant metastases received systemic chemotherapy, and 30 (19%) received palliative therapy. RESULTS: Excluding the CR group, the patients in the RR group had a higher 1-year survival rate than the other treatment groups. However, there was no significant difference in the overall survival rates of the RR, RT, and TACE/TAI groups. When we evaluated prognostic factors to clarify the indications for RR in the multidisciplinary treatment of patients with advanced HCC with TT, prothrombin activity (PA) was identified as a significant independent preoperative factor for overall survival in the RR group. The survival rate in patients with PA of < or = 78% was significantly lower than that of patients with PA of > 78% (P = 0.0004). The median survival time of patients with serum PA of > 78% who underwent RR was 13.9 months and that of patients who underwent CR was 9.1 months, with no survival difference between the groups. CONCLUSION: In advanced HCC with major vascular involvement, patients who had RR with PA of greater 78% achieved a similar survival to those who had CR. The surgeon should still proceed with RR in those patients with serum PA of > 78% if CR does not seem feasible on preoperative evaluation. PMID- 16479352 TI - Doppler Adson's test: predictor of outcome of surgery in non-specific thoracic outlet syndrome. AB - INTRODUCTION: Outcome of surgery in non-specific thoracic outlet syndrome (TOS) is variable. Duplex imaging of the subclavian artery during Adson's maneuvre may help predict outcome. PATIENTS AND METHODS: A retrospective study of 16 cases of non-specific TOS who had pre-operative duplex imaging of the subclavian artery during Adson's maneuvre was done and their outcomes following surgical decompression of the thoracic outlet studied. RESULTS: Of the eight cases with a positive Adson's test, seven (87.5%) had a complete response of symptoms following surgery while in the eight cases with a negative test, only four (50%) had a favorable response to surgery. CONCLUSION: Duplex scanning of the thoracic outlet during Adson's maneuvre may help predict outcome of thoracic outlet decompression surgery in cases of non-specific TOS. PMID- 16479353 TI - Preoperative nutritional risk assessment in predicting postoperative outcome in patients undergoing major surgery. AB - INTRODUCTION: Although a variety of nutritional indices have been found to be valuable in predicting patient outcome when used alone, there is no consensus on the best method for assessing the nutritional status of hospitalized patients. Therefore, the aim of this study was to assess the nutritional status of a cohort of patients who underwent major elective surgery using the Nutritional Risk Index (NRI), Maastricht Index (MI), Subjective Global Assessment (SGA), and Mini Nutritional Assessment (MNA) to determine the best possible nutrition screening system in surgical practice. METHODS: The study population consisted of 460 patients who underwent major elective surgery between December 1999 and March 2002. Each patient had a complete set of the three nutritional assessment techniques (NRI, MI, SGA); in addition, the MNA was performed in patients older than 59 years of age. One of the coauthors who was unaware of the nutritional assessments assessed the patients for postoperative morbidity and mortality. Complications were classified as major or minor and as infectious or noninfectious. To assess the predictive value of the assessment techniques, likelihood ratios were calculated for the various strata of each method. The odds ratio and receiver operating characteristic (ROC) curves were also calculated to describe and compare the diagnostic value of each of the four nutrition indices. RESULTS: Twenty patients died during the study period. No complications occurred in 329 of the 460 patients; 42 patients suffered from two or more complications. The frequency of malnutrition was found to be 58.3%, 63.5%, and 67.4% as assessed by the SGA, NRI, and MI, respectively. Morbidity rates, especially severe infectious and noninfectious complications, were significantly higher in malnourished patients in all nutritional indices. The likelihood ratio was well correlated with the risk categories of every nutritional index. The area under the ROC curves revealed that each scoring system proved to be significantly powerful in predicting the morbidity (infectious and noninfectious severe morbidity) and mortality. However, no differences were detected among the nutritional indices in 460 patients. The odds ratio for morbidity between the well nourished and malnourished patients was 3.09 [95% confidence interval (CI), 1.96-4.88], 3.47 (95% CI, 2.12-5.68), 2.30 (95% CI, 1.43-3.71), and 2.81 (95% CI, 0.79-9.95) for the SGA, NRI, MI, and MNA, respectively. All indices except the MNA were significantly predictive for morbidity. The odds ratios were not statistically different among the indices. CONCLUSIONS: Our findings revealed that all nutritional assessment techniques can be safely applied to the clinical setting with no significant difference in predictive value. We therefore strongly recommend the use of any of these techniques to improve the outcome of surgical care. Meanwhile, further investigations are needed, and much effort must be given to find the best method for assessing nutritional status. PMID- 16479354 TI - Interval appendectomy after conservative treatment of an appendiceal mass. AB - INTRODUCTION: The purpose of this study was to clarify the role of interval appendectomy after conservative treatment of an appendiceal mass. METHODS: From January 1998 to December 2003, patients with an appendiceal mass who received conservative treatment at the Taipei Veterans General Hospital were studied retrospectively. Data on demographics, rate of appendicitis recurrence, duration of hospital stay, and complication rate were collected and analyzed. RESULTS: A total of 165 patients were included (89 males, 76 females). The mean age was 53.6 years (range 7-89 years). The rate of appendicitis recurrence after conservative treatment was 25.5%; most recurred within 6 months after discharge (83.3%). The benefit of preventing recurrence is less than 16% if interval appendectomy is performed 6 weeks after discharge and less than 10% if it is done 12 weeks later. The complication rate of appendectomy performed before or after recurrence was 10% in both groups. The duration of the second hospital stay for patients who underwent interval appendectomy before or after recurrence was 4.43 +/- 3.32 vs. 6.75 +/- 5.73 days (P = 0.023). Of the 165 patients, 17 (10.3%) had their diagnosis changed after survey or surgery, and 5 (3.03%) were found to have colon cancer upon follow-up. CONCLUSIONS: Patients who recovered from conservative treatment of an appendiceal mass should undergo colonoscopy or barium enema to detect any underlying diseases and to rule out coexistent colorectal cancer. Routine interval appendectomy benefits less than 20% of patients. PMID- 16479355 TI - Glucose metabolism in batch and continuous cultures of Gluconacetobacter diazotrophicus PAL 3. AB - Periplasmic glucose oxidation (by way of a pyrrolo-quinoline-quinone [PQQ]-linked glucose dehydrogenase [GDH]) was observed in continuous cultures of Gluconacetobacter diazotrophicus regardless of the carbon source (glucose or gluconate) and the nitrogen source (N(2) or NH(3)). Its synthesis was stimulated by conditions of high energetic demand (i.e., N(2)-fixation) and/or C-limitation. Under C-excess conditions, PQQ-GDH synthesis increased with the glucose concentration in the culture medium. In batch cultures, PQQ-GDH was actively expressed in very early stages with higher activities under conditions of N(2) fixation. Hexokinase activity was almost absent under any culture condition. Cytoplasmic nicotinamide adenine dinucleotide (NAD)-linked glucose dehydrogenase (GDH) was expressed in continuous cultures under all tested conditions, and its synthesis increased with the glucose concentration. In contrast, low activities of this enzyme were detected in batch cultures. Periplasmic oxidation, by way of PQQ-GDH, seems to be the principal pathway for metabolism of glucose in G. Diazotrophicus, and NAD-GDH is an alternative route under certain environmental conditions. PMID- 16479356 TI - Dissimilatory nitrate reductase from Bradyrhizobium sp. (Lupinus): subcellular location, catalytic properties, and characterization of the active enzyme forms. AB - Subcellular location, chlorate specificity, and sensitivity to micromolar concentrations of azide suggest that most of the anaerobically induced nitrate reductase (NR) activity in Bradyrhizobium sp. (Lupinus) could be ascribed to the membrane type of bacterial dissimilatory NRs. Two active complexes of the enzyme, NR(I) of 140 kDa and NR(II) of 190 kDa, were detected in membranes of the nitrate respiring USDA strain 3045. Both enzyme forms were purified to homogeneity. Obtained specific antibodies showed that these native species were immunologically closely related and composed of largely similar 126-kDa, 65-kDa, and 25-kDa subunits. The finding that NR(I) and NR(II) share common epitopes suggests that they may not be different species, but rather two forms of the same enzyme. PMID- 16479357 TI - Biotransformation of 1-naphthol by a strictly aquatic fungus. AB - The aquatic hyphomycete Heliscus lugdunensis belongs to a group of exclusively aquatic mitosporic fungi with an only scarcely explored potential to oxidatively attack xenobiotic compounds, and was used to study the biotransformation of the environmental pollutant metabolite 1-naphthol. H. lugdunensis metabolized approximately 74% of 1-naphthol within 5 days. The identification and quantification of degradation products using gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, and high performance liquid chromatography revealed that approximately 12% of the parent compound was converted into 1-naphthylsulfate, 3% was transformed into 1-methoxy-naphthalene, and less than 1% was converted into 1,4-naphthoquinone. A further metabolite, most likely 4-hydroxy-1-naphthylsulfate, was also detected. In contrast to sulfate conjugate metabolites, no glucuronide and glucoside conjugates of 1 naphthol were found, and neither UDP-glucuronyltransferase nor UDP glucosyltransferase present in H. lugdunensis showed activity towards 1-naphthol. These results support a role of fungi adapted to aquatic environments in affecting the environmental fate of pollutants in aquatic ecosystems. PMID- 16479358 TI - Construction of pDESTR, a GATEWAY vector for gene disruption in filamentous fungi. AB - We have constructed pDESTR, a destination vector of gateway system especially for gene targeting and disruption in filamentous fungi. The vector was constructed by removing the multicloning site of pGEM-T easy vector, and inserting hygromycin phosphotransferase gene construct from pCB1004, and a gateway vector conversion cassette. In order to construct a DNA for gene disruption, only an inverse polymerase chain reaction (PCR) amplification of the restricted, target sequence is needed. After the amplification with a 5'CACC-tagged primer and an ordinary primer, the DNA fragment will be inserted into pENTR/D-TOPO vector and then transferred into pDESTR through LR-recombination reaction. The resulting vector has the disruption construct, after being digested with the restriction enzyme used for the inverse-PCR. The effectiveness of this vector was assessed in Neurospora crassa. The use of pDESTR will therefore simplify the construction of a targeting vector, where multiple ligation steps are usually needed. PMID- 16479359 TI - Oligonucleotide microarray with RD-PCR labeling technique for detection and typing of human papillomavirus. AB - Currently, screening for high-risk human papillomavirus (HPV) infection remains an important health concern throughout the world, because of the close association between certain types of HPV and cervical cancer. In this study, we explore the possibility of using approximately 70mer oligonucleotide microarray for detection and genotyping of HPV. The approximately 70mer type-specific oligonucleotide probes of four different types HPV were designed by using biological software Array designer 2.0, which analyzed the whole genome sequences of HPV and selected optimal probes. These probes were synthesized and printed onto the surface of glass slides in order to prepare a low-density microarray. HPV samples were labeled with fluorescence dyes Cy3 using a method of restriction display polymerase chain reaction (RD-PCR). HPV plasmid DNA was restricted with Sau3A I to produce multiple fragments that were ligated to adaptors subsequently and used as PCR template. PCR labeling was performed with the fluorescently labeled universal primer (Cy3-UP) whose sequence is designed according to the adaptor of the RD-PCR approaches. The labeled samples were hybridized with the oligonucleotide microarray. The scanning results showed that HPV DNA hybridized specifically with multiple spots correspondingly to show positive signals, whereas no signals were detected of all the negative and blank controls. These results demonstrated that approximately 70mer oligonucleotide microarray can be applied to HPV detection and genotyping. The application of RD-PCR in the sample labeling can increase significantly the sensitivity of the assay and will be especially useful for the discriminate diagnosis of multiple pathogens. PMID- 16479360 TI - Differences in ocular dimensions between normal and dry eyes. AB - Assessment of ocular dimensions is essential for ophthalmic surgeons because these values must be determined before scheduling excimer laser refractive and cataract surgeries. Dry eye seems to affect central corneal thickness (CCT) values, but it is not clear if it affects anterior chamber depth (ACD), lens thickness (LT), vitreous chamber depth (VCD) and axial length values. Following on from this, we measured the CCT, ACD, LT, VCD and axial length of 64 healthy eyes (51.20%) and 61 dry eyes (48.80%). CCT was measured with scanning-slit corneal topography (Orbscan Topography System II, Orbscan, Inc., Salt Lake City, UT, USA) and ACD, LT, VCD and axial length with a 10-MHz A-mode ultrasound device (Compuscan; Storz, St. Louis, MO, USA). There were no significant differences in ACD (P=0.588), LT (P=0.739), VCD (P=0.568) and axial length (P=0.199) between normal and dry eyes. Nevertheless, the differences in CCT between normal (549+/ 34 microm) and dry eyes (527+/-30 microm) were significant (P<0.001). In sum, it seems that only the CCT values are significantly lower in subjects with dry eye. PMID- 16479361 TI - A rich lymphatic network exists in the inferior surface of the vocal cord. AB - Conservation laryngeal surgery is an increasingly available alternative for the treatment of laryngeal cancer. Understanding anatomy of laryngeal lymph drainage is essential for clinicians to diagnose, grade and surgically manage the laryngeal cancer. Although the lymphatic drainage of the larynx has been extensively studied, few studies revealed the relationship of the lymphatic drainage between various parts of the larynx. The distribution of lymphatic vessels in the inferior surface of the vocal cord also remains unclear. The aim of this study was to investigate the communication of the lymphatic networks between the vocal cord, the supraglottic and subglottic parts of the larynx, paying special attention to the lymphatic drainage of the inferior surface of the vocal cord. Eighteen larynx specimens from 18 fresh fetal cadavers were manually injected with prassion blue solution into the mucosal or submucosal layer of the larynx in order to reveal the lymphatic vessels in the inner larynx. We found that a rich lymphatic network is present in the inferior surface of the vocal cord, and the lymphatic networks in the superior and inferior surfaces of the vocal cord appear as two different patterns. These findings provide an anatomical basis for the design of a partial or conservation laryngeal surgery, particularly when considering the precise resection margin. PMID- 16479362 TI - Laparoscopic total extraperitoneal repair versus anterior preperitoneal repair for inguinal hernia. AB - Laparoscopic inguinal hernia repair is still not the gold standard for repair although mesh implantation is unequivocally accepted as an integral part of any groin hernia repair. The aim of the study was to compare the results of anterior preperitoneal (APP) mesh repair with totally extra peritoneal (TEP) repair for inguinal hernias. The prospective study was conducted on 241 patients with 247 hernias (from January 2000 to June 2004). Anterior preperitoneal repair was done in 121 patients and 120 patients were subjected to TEP repair. Repair in both groups was done by using Prolene mesh of size 6x4 in. or 6x6 in. intraoperative and postoperative parameters and complications were recorded and the patients were followed up to 1 year post-surgery. For both unilateral and bilateral inguinal hernias, mean operative time was significantly more in patients of TEP repair as compared to APP repair (P<0.001) and significantly more patients had peritoneal tears in the TEP group (P<0.001). Patients undergoing TEP repair, however, had significantly less postoperative pain (P<0.05) and postoperative hospital stay (P<0.05) and return to work was significantly earlier is this group (P<0.01 and P<0.001). There was no difference in the recurrence rate between the two groups. Patients with inguinal hernias undergoing laparoscopic repair recover more rapidly, and have less incidence of postoperative pain. But it takes significantly more time to perform than APP repair and also the incidence of peritoneal tear is higher. PMID- 16479363 TI - Remembering Lee McIntosh (1949-2004), a pioneer in the molecular biology of chloroplast and mitochondrion function. PMID- 16479367 TI - Urgent colectomy and caesarean section of a pregnant familial adenomatous polyposis: a case report. PMID- 16479369 TI - Laser-induced breakdown spectroscopy (LIBS) for carbon single shot analysis of micrometer-sized particles. AB - The purpose of this work is to study the ability of the laser-induced breakdown spectroscopy (LIBS) technique to perform in situ (without sample preparation) detection of graphite particles circulating in a gas loop used to simulate the cooling gas circuit of a helium-cooled nuclear reactor. Results obtained with a laboratory scale set up are presented. The experiments were performed in nitrogen with micrometer-sized particles containing carbon (glucose particles and sodium hydrogenocarbonate particles). Statistical shot to shot analysis was used to determine the concentration of the analyte. The variation of LIBS signal as a function of glucose particle diameter showed an underestimation of the signal of particles of diameters larger than 5 microm. This phenomenon is likely to be correlated to an incomplete vaporization in the laser-induced plasma of particles of sizes above 5 microm. Analytical measurements were performed with glucose particles and sodium hydrogenocarbonate particles, and the concentration-based limit of detection of carbon was evaluated to be about 60 microg m(-3). PMID- 16479368 TI - Stapled vs open hemorrhoidectomy: long-term outcome of a randomized controlled trial. AB - BACKGROUND AND AIMS: Stapled hemorrhoidectomy is a relatively new procedure, and studies on long-term outcomes are few. We present the results of a 5-year follow up on patients recruited to a randomized controlled trial comparing stapled and Milligan-Morgan hemorrhoidectomy. PATIENTS AND METHODS: We performed telephone interviews and office visits between May and July 2005 on patients who had taken part in a randomized controlled trial from May 1999 to December 2000. RESULTS: Occasional pain and bleeding were referred by the patients with no difference between the two groups. The patients were also equally satisfied with both procedures. No recurrent hemorrhoidal prolapse or stenosis was detected at anorectal exploration and rigid sigmoidoscopy. CONCLUSION: Both stapled and Milligan-Morgan techniques guarantee satisfactory long-term results. Larger studies are needed to assess the durability of stapled hemorrhoidectomy. PMID- 16479370 TI - Differential proteome analysis of tonsils from children with chronic tonsillitis or with hyperplasia reveals disease-associated protein expression differences. AB - A proteomic approach has been used to establish a proteome map and differentiate between the protein composition of tonsils from patients with chronic tonsillitis (CT) and that of tonsils with hyperplasia (HPL). Two-dimensional gel analysis was performed with material from four patients with HPL and five patients with CT. An average of approximately 600 spots were detected in each gel. A total of 127 different proteins were identified in 158 spots analyzed by mass spectrometry. Our study revealed disease-associated differences between protein abundance for two protein spots, an HSP27 isoform and UMP-CMP kinase. Both protein spots were more abundant in the CT group. HSP27 ELISA was performed for 32 patients, 12 belonging to the HPL group and 20 to the CT group. ELISA could not be used to differentiate HSP27 isoforms nor to distinguish CT from HPL. HSP27 was found to migrate to two further protein spots in the 2D gels. The differently expressed HSP27 isoform migrated as the most acidic of all the HSP27 isoforms detected, indicating the highest degree of phosphorylation. The sum of all three HSP27 abundances in the gels from the CT group was not different from that of the HPL group, consistent with the ELISA results. Our results suggest that phosphorylation differences caused the observed migration differences of HSP27. Together with the UMP-CMP kinase abundance differences, we conclude that kinase and/or phosphatase activity are different in CT and HPL. PMID- 16479371 TI - Flow injection chemiluminescence immunoassay for 17beta-estradiol using an immunoaffinity column. AB - A new flow injection chemiluminescent immunoassay was developed for the detection of 17beta-estradiol (E2). The method uses p-iodophenol (PIP) as enhancer and is based on a solid-phase immunoassay format in which an E(2)-OVA immobilized immunoaffinity column inserted in the flow system is used to trap unbound horseradish peroxidase (HRP)-labeled anti-E2 antibody after an off-line incubation of E2 with HRP-labeled anti-E2 antibody. The trapped enzyme conjugate was detected by injecting substrates to produce an enhanced chemiluminescence (CL) response. The linear range for E2 was 10.0-1,000.0 ng mL(-1) with a correlation coefficient of 0.996 and a detection limit of 3.0 ng mL(-1). The sampling and chemiluminescence detection time for one sample was 400 s after a pre-incubation procedure of 30 min. Serum samples detected by this method were in good agreement with the results obtained by EIA with E2-biotin. PMID- 16479372 TI - Effects of anti-phencyclidine and anti-(+)-methamphetamine monoclonal antibodies alone and in combination on the discrimination of phencyclidine and (+) methamphetamine by pigeons. AB - RATIONALE: Drug-specific monoclonal antibodies against phencyclidine (PCP) and (+)-methamphetamine [(+)-METH] should bind to these drugs to block their discriminative stimulus effects. OBJECTIVES: To determine if mouse monoclonal antibodies against PCP and (+)-METH can block the discriminative stimulus effects of the drugs in pigeons. MATERIALS AND METHODS: Pigeons were trained to discriminate among intramuscular injections of saline, 1 mg/kg PCP, and 2 mg/kg (+)-METH. After responding stabilized, cumulative dose-response curves were obtained for PCP and (+)-METH. Doses of an anti-PCP antibody at 620 mg/kg (anti PCP mAb6B5) with a K (D) of 1.3 nM for PCP and no measurable affinity for (+) METH and 1,000 mg/kg doses of anti-(+)-METH antibody (anti-METH mAb6H7) with a K (D) of 41 nM for (+)-METH and no measurable affinity for PCP were subsequently administered, first alone and later in combination after which the dose-response curves were redetermined. RESULTS: When the antibodies were given alone, the anti PCP antibody blocked the discriminative stimulus effects of PCP, but not those of (+)-METH, and the anti-(+)-METH antibody blocked the discriminative stimulus effects of (+)-METH, but not those of PCP. The anti-PCP antibody shifted the PCP dose-response curve further to the right and for a longer time than the anti-(+) METH antibody shifted the dose response curve for (+)-METH. When the anti-PCP and anti-(+)-METH antibodies were administered on the same day, the discriminative stimulus effects of both drugs were completely blocked 1 day after antibody administration. CONCLUSIONS: These experiments demonstrate the high specificity of the antibodies for the drugs to which they bind and show that monoclonal antibodies can be combined to antagonize the effects of more than one drug. PMID- 16479373 TI - Pharmacological characterisation of place escape/avoidance behaviour in the rat chronic constriction injury model of neuropathic pain. AB - RATIONALE: Classical pain tests performed in animals routinely measure evoked nociceptive behaviours. These almost exclusively reflect sensory processing of nociceptive transmission, although a recently described place escape/avoidance paradigm may be used to selectively assess affective pain processing. OBJECTIVE: To establish if drugs with proven analgesic efficacy selectively attenuate sensory-discriminative or affective-motivational aspects of nociceptive processing. METHODS: The mu-opioid receptor agonist morphine, the anti-epileptic gabapentin, the anti-depressant duloxetine, the 5HT1A receptor agonist 8-OH-DPAT, the GABA(A) receptor agonist gaboxadol and the mixed cannabinoid receptor agonist WIN55,212-2 were tested after systemic administration in the chronic constriction injury (CCI) model of neuropathic pain. For the place escape/avoidance paradigm, CCI rats had free access between the 'non-aversive' dark and 'aversive' light side of an enclosed chamber. Either the injured or non-injured hindpaw was routinely stimulated if the rat was in the dark or light area, respectively. Escape/avoidance behaviour was defined as a shift from the dark to the light area. Mechanical allodynia and hyperalgesia were determined prior to and following escape/avoidance testing. RESULTS: Morphine (3 and 6 mg/kg), gabapentin (50 and 100 mg/kg), duloxetine (10 and 30 mg/kg) and 8-OH-DPAT (0.1 and 0.5 mg/kg) attenuated the time spent by CCI rats in the light area; gaboxadol (1 and 3 mg/kg) and WIN55,212-2 (0.3 and 1 mg/kg) were ineffective. Only gabapentin and 8-OH-DPAT attenuated mechanical nociceptive behaviours at non-sedative doses. CONCLUSIONS: The place escape/avoidance paradigm may enable discrimination between selected drug classes on distinct components of sensory and affective pain processing in rats with neuropathic pain. PMID- 16479374 TI - Ultrastructure of the spermatozoon of Dollfusiella spinulifera (Beveridge and Jones, 2000) Beveridge, Neifar and Euzet, 2004 (Trypanorhyncha, Eutetrarhynchidae). AB - The ultrastructural organization of the mature spermatozoon of the trypanorhynch cestode Dollfusiella spinulifera is described. The spermatozoon is a long filiform cell, tapered at both ends, which lacks mitochondria. Its cytoplasm contains (1) two axonemes of different lengths of the 9+'1' pattern of trepaxonematan Platyhelminthes, (2) two rows of parallel cortical microtubules, each of which is adjacent to the opposite sides of the plasma membrane, (3) the nucleus, and (4) glycogen in the form of both alpha-glycogen rosettes and beta glycogen particles. Unlike the majority of cestode spermatozoa, the spematozoon of D. spinulifera lacks crested bodies and periaxonemal sheath. In view of the present results, the postulated synapomorphic value of crested bodies for the Eucestoda is questioned. PMID- 16479375 TI - Molecular cloning and characterization of a novel lactate dehydrogenase gene from Clonorchis sinensis. AB - From a Clonorchis sinensis adult worm cDNA library, we isolated a cDNA clone encoding a novel lactate dehydrogenase (LDH) gene which encoded a putative protein with a predicted molecular weight of 35.6 kDa. The optimum pH and temperature for the enzyme were 7.5 and 50 degrees C in the pyruvate reduction while 11 and 80 degrees C in the lactate oxidation reaction, respectively. CsLDH showed no substrate inhibition by high lactate and NAD(+) concentration, and the optimal pyruvate and optimal NADH concentrations were 10 and 0.5 mmol/l, respectively. The relative activities of these 2-oxocarboxylic acids were pyruvic acid>2-ketobutyrate>oxalacetic acid>alpha-ketoglutaric acid>phenylpyruvate. The cofactor 3-acetylpyridine adenine dinucleotide was much more effective than NAD(+). The cofactor analogs in which the nicotinamide ring is replaced by 3 pyridinealdehyde were lower activity cofactors, while the nicotinamide ring is replaced by nicotinic acid or thionicotinamide which is not a cofactor to CsLDH. The succinic acid and malic acid are not substrates of CsLDH. Cu(2+), Fe(2+), and Zn(2+) greatly inhibited the CsLDH activity both in the direction of pyruvate reduction and in the direction of lactate oxidation. The inhibition of CsLDH by gossypol may make gossypol a potential therapy drug or a lead compound for C. sinensis. Accordingly, the CsLDH may be a novel potential drug target. PMID- 16479376 TI - Accumulation of tropomyosin isoform 5 at the infection sites of host cells during Cryptosporidium invasion. AB - The actin cytoskeleton of host cells has been implicated in Cryptosporidium invasion. However, the underlying mechanism of how actin filaments and associated proteins modulate this process remains unclear. In this study, we use in vitro cultured cell lines, human ileocecal adenocarcinoma HCT-8 and Chinese hamster ovary (CHO), and an in vivo mouse model to investigate the roles of tropomyosin isoforms in Cryptosporidium invasion. Using isoform-specific monoclonal antibodies, we found that the major human tropomyosin (hTM) isoforms expressed in HCT-8 cells are hTM4 and hTM5. HCT-8 cells also express hTM1 at low levels but not hTM2 and hTM3. During Cryptosporidium parvum infection, hTM5 colocalized to the infection sites with a novel parasite membrane protein, CP2. Neither hTM1 nor hTM4 accumulated at infection sites. Similarly, a high level of TM5 and varying amounts of TM4 accumulated at the C. parvum infection sites in CHO cells. CHO cells overexpressing hTM5 exhibit a significantly higher percent of mature meronts early in the infection process relative to CHO cells or CHO cells overexpressing a tropomyosin mutant, chimeric isoform hTM5/3. These results suggest that functional TM5 enhances Cryptosporidium invasion of host cells. In C. parvum-infected mice, accumulation and rearrangement of TM5 and TM4 were detected throughout the infected ileum. Similarly, in the Cryptosporidium muris infected mice, TM5 accumulated in discrete regions of the epithelial cells of gastric glands and in the oocyst-laden stomach gland lumen. Cryptosporidium infection appears to rearrange and recruit host TM isoforms in both culture cells and in the mouse. Localized accumulation of tropomyosin at the infection sites may facilitate parasite invasion. PMID- 16479379 TI - Comment on "Changes in acid-base balance following bolus infusion of 20% albumin solution in humans" by Bruegger et al. PMID- 16479377 TI - The changing pathogens of complicated parapneumonic effusions or empyemas in a medical intensive care unit. AB - OBJECTIVES: To assess the incidence, pathogens, and outcome of complicated parapneumonic effusions or empyemas in a medical intensive care unit (MICU) patients with pleural effusions. DESIGN AND SETTING: Prospective study of febrile MICU patients with pleural effusion carried out in a tertiary care hospital between April 2001 and September 2003. PATIENTS: The study included 175 patients with a temperature above 38 degrees for more than 8 h with evidence of pleural effusion confirmed by chest radiography and ultrasound. INTERVENTION: Routine thoracentesis and effusion cultures. RESULTS: The prevalence of complicated parapneumonic effusions or thoracic empyemas in febrile MICU patients with pleural effusions was 45% (78/175). A total of 78 micro-organisms were isolated from the pleural fluid of 58 patients (positive microbiological culture 74%) including aerobic Gram-negative (n=45), aerobic Gram-positive (n=23), anaerobic (n=5), Myobacterium tuberculosis (n=3), and Candida (n=2). The infection-related mortality rate of complicated parapneumonic effusions or empyemic patients in the MICU was 41% (32/78). CONCLUSION: The development of complicated parapneumonic effusions or thoracic empyemas in MICU patients is a high-mortality disease. The increasing incidence of aerobic Gram-negative pathogens in empyema has become a more urgent problem. PMID- 16479380 TI - Organophosphate poisonings with parathion and dimethoate. AB - OBJECTIVE: Organophosphate toxicity is the leading cause of morbidity and death in poisoning by insecticides. The clinical symptoms of pesticide toxicity range from the classic cholinergic syndrome to flaccid paralysis and intractable seizures. The mainstays of therapy are atropine, oximes, benzodiazepines and supportive care. The toxicokinetics vary not only with the extent of exposure, but also with the chemical structure of the agent. PATIENTS: We report two cases of poisoning with parathion-ethyl and dimethoate. The patients developed a cholinergic syndrome immediately, accompanied by bradycardia and hypotension. INTERVENTIONS: The patients were admitted to the intensive care unit (ICU) a few hours after ingestion. Atropine was administered according to the cholinergic symptoms. The patients recovered in the ICU after 10-12 days and were discharged after 3 and 4 weeks. MEASUREMENTS AND RESULTS: Organophosphate blood and urine levels were determined on admission and during hospitalisation. The pesticides were rapidly distributed and slow elimination rate of the poisons was documented. In the case of parathion-ethyl the distribution half-life estimated was t(1/2alpha) = 3.1h while the terminal half-life was t(1/2beta) = 17.9 h. Using a one-compartment model for dimethoate the elimination half-life was t(1/2beta) = 30.4 h in plasma and 23.8 h in urine. The serum pseudo-cholinesterase activity was below the limit of detection at admission and recovered during the following 3weeks. PMID- 16479381 TI - Relationship between pressure-volume curve and markers for collagen turn-over in early acute respiratory distress syndrome. AB - OBJECTIVE: In acute respiratory distress syndrome, the relationships between changes in the elastic behavior of the respiratory system and biological markers of extra-cellular matrix or surfactant turn-over could give some insights into its pathophysiological determinants. DESIGN AND MEASUREMENTS: In 17 patients with acute respiratory distress syndrome, we assessed the relationship between chord compliance measured on pressure-volume curves obtained at two levels of positive end-expiratory pressure (0 and 10[Symbol: see text]cm[Symbol: see text]H(2)O) and biological markers of collagen turn-over or surfactant degradation in bronchoalveolar lavage fluid obtained simultaneously in the early phase of the disease (first 4 days). MAIN RESULTS: The compliance of the respiratory system obtained from the pressure-volume curves was significantly correlated with markers for collagen turn-over (type III procollagen peptide and matrix metalloproteinase 2) and with markers of surfactant degradation (type-IIA secretory phospholipase A2). The correlations were stronger when the curve was traced from positive end-expiratory pressure, suggesting that this condition may improve the assessment of tissue mechanics. A logarithmic relationship best described the correlation between compliance and type III procollagen peptide, in agreement with a collagen-dependent model of maximal distension. The marker for surfactant degradation was associated with ongoing alveolar inflammation (cellularity of bronchoalveolar lavage fluid and tumor necrosis factor-alpha concentration). Interleukin-10, an anti-inflammatory mediator, showed no correlation with compliance. CONCLUSION: These preliminary data suggest that a severe reduction in compliance in the early phase of acute respiratory distress syndrome is associated with both collagen deposition and surfactant degradation. PMID- 16479382 TI - An evaluation of systemic inflammatory response syndrome signs in the Sepsis Occurrence In Acutely Ill Patients (SOAP) study. AB - OBJECTIVE: To define the frequency and prognostic implications of SIRS criteria in critically ill patients hospitalized in European ICUs. DESIGN AND SETTING: Cohort, multicentre, observational study of 198 ICUs in 24 European countries. PATIENTS AND INTERVENTIONS: All 3,147 new adult admissions to participating ICUs between 1 and 15 May 2002 were included. Data were collected prospectively, with common SIRS criteria. RESULTS: During the ICU stay 93% of patients had at least two SIRS criteria [respiratory rate (82%), heart rate (80%)]. The frequency of having three or four SIRS criteria vs. two was higher in infected than non infected patients (p < 0.01). In non-infected patients having more than two SIRS criteria was associated with a higher risk of subsequent development of severe sepsis (odds ratio 2.6, p < 0.01) and septic shock (odds ratio 3.7, p < 0.01). Organ system failure and mortality increased as the number of SIRS criteria increased. CONCLUSIONS: Although common in the ICU, SIRS has prognostic importance in predicting infections, severity of disease, organ failure and outcome. PMID- 16479383 TI - ["One shot" CO2 laser stapedotomy]. AB - BACKGROUND: In order to further optimize the surgical technique with CO(2) laser in stapes surgery, a scanner system was used to obtain a footplate perforation of 0.5-0.6 mm with only one laser application ("one-shot" stapedotomy). PATIENTS AND METHODS: A total of 255 patients with otosclerosis were submitted to primary CO(2) laser stapedotomy with a SurgiTouch scanner. This study discusses the surgical technique and clinical results. RESULTS: An adequately large perforation diameter could be achieved with a single shot in 68% of the patients treated. In 14% of the patients, a second laser application at the same site was necessary. In 18% the perforation had to be enlarged by several slightly overlapping laser applications without using the scanner. There was no evidence of laser dependent inner ear affections. CONCLUSION: CO(2) laser, combined with modern scanner systems, is well suited for application in stapes surgery. PMID- 16479384 TI - [Transoral resection of locally advanced squamous cell carcinoma of the lateral oropharynx: combination of CO2 laser microsurgery with flap reconstruction]. AB - INTRODUCTION: Transoral laser microsurgery of locally advanced carcinomas of the lateral pharynx often results in exposure of major vessels of the neck and is accompanied by a substantial risk of intra- and postoperative bleeding. We therefore only perform these operations after external protection of neck vessels, if necessary combined with flap reconstruction. PATIENTS AND METHODS: Between October 2001 and December 2004, 11 locally advanced squamous cell carcinomas of the lateral oropharynx that reached the major vessels of the neck were treated as follows: after ipsilateral neck dissection with temporary protection of the jugular vein and carotid arteries, the neck remained open while transoral laser surgery of the primary tumor was performed. Pharyngeal defects were subsequently closed by either primary suture or a platysma myofascial flap. All patients underwent adjuvant radiotherapy. RESULTS: All primary tumors were completely resected. None of the patients required tracheotomy or placement of a percutaneous endoscopic gastrostomy tube. The mean duration of nasogastric feeding tubes was 12.7 days. In one case, the routine radiological contrast study revealed a blind cervical fistula 10 days after surgery. This healed spontaneously within 7 days. One mild postoperative hemorrhage had to be stopped by endoscopic coagulation under general anesthesia. During a mean follow-up of 19.4 months, none of the patients developed a local and/or regional recurrence. CONCLUSION: The surgical procedure described ensures sufficient protection of neck vessels during and after the transoral resection of advanced carcinomas of the pharynx. It successfully combines the advantages of minimally invasive laser microsurgery with those of flap reconstruction known from traditional surgery. PMID- 16479385 TI - [Optical coherence tomography in the diagnosis of vocal folds]. AB - Optical coherence tomography (OCT) is a new, non-invasive method for investigating biological tissue, with a beam penetration of up to 3 mm. In this study, OCT was used on porcine and human larynxes. The porcine vocal folds were assigned to defined areas and examined by OCT followed by traditional histomorphological analysis. We were able to validate this new method by demonstrating that both OCT and the histological assays showed a clear demarcation of the epithelium mucosae from the deeper layers. OCT application to human vocal folds delivered same results. The current standard procedure for ensuring an accurate diagnosis if tissue is malignant, is still an invasive one. Our results show that OCT allows the collection of crucial information on the quality of morphological changes on a vocal fold without the necessity of being invasive. Thus, we propose that OCT be introduced as a new method for the detection of morphological changes in the vocal fold complementary to other established methods. PMID- 16479386 TI - [Influence of ischemia/hypoxia on the HIF-1 activity and expression of hypoxia dependent genes in the cochlea of the newborn rat]. AB - BACKGROUND: Transcription factor HIF-1 (hypoxia-inducible factor-1) regulates the expression of genes which are involved in glucose supply, growth, metabolism, redox reactions and blood supply. Hypoxia and ischemia play an important role in the pathogenesis of tinnitus and hearing loss. Therefore, HIF-1 activity and the expression of HIF-1 dependent genes in the cochlea were examined under ischemic and hypoxic conditions. MATERIAL AND METHODS: For the HIF-1 analysis, single-cell cultures of the organ of Corti (OC), stria vascularis (SV) and modiolus (MOD) were used. mRNA expression was analyzed in the organotypic culture using a microarray technique (RN U34-chip, Affymetrix). RESULTS: Ischemia (hypoxia without glucose) and pure hypoxia increase the HIF-1 activity identically, with the highest increase found in MOD and OC. The HIF-1 alpha mRNA levels were found to be higher in SV than in the OC and MOD. During culturing, there is a clear increase in HIF-1 alpha mRNA and the expression of a number of HIF-1 dependent genes, such as Gapdh/glyceraldehyde-3-phosphate dehydrogenase, Slc2a1/solute carrier family 2 (facilitated glucose transporter), member 1, Tf/transferrin and Tfrc/transferrin receptor, in all three regions. In SV, MOD and OC, increase in the expression of Hmox1/hemoxygenase 1, Nos2/nitric oxide synthase, inducible and Tfrc is particularly high. Hypoxia (5 h) results in an increased expression of Igf2/Insulin-like growth factor 2. CONCLUSION: The present data underline the contribution of radical forming processes to the pathogenesis of inner ear diseases. For experimental research, it is important to note that organotypic culture may be coupled with hypoxia. PMID- 16479387 TI - [Solitary intraparotid neurofibroma of the facial nerve. Symptomatology, biology and management]. AB - Neurogenic neoplasms of the parotid gland are extremely uncommon. We present the case of a solitary intraparotid neurofibroma of the N. facialis occurring in a 55 year-old female who noted a painless left-side enlargement in the region of the parotid gland over a period of 2 years. Facial function was normal. Magnetic resonance imaging revealed a well-demarcated round mass within the left parotid. At surgery, a tumor was found involving the main trunk of the facial nerve, histopathologically representing a neurofibroma. Since solitary intraparotid neurofibromas are characterized by a very slow growth lacking the propensity for malignant transformation, the tumor was left to preserve facial nerve function. The clinical course over months or even years and the non-specific symptomatology are characteristic of intraparotid facial neurofibromas. Surgical management depends on the clinical setting in the individual case. Conservative treatment based on facial nerve preservation and leaving the tumor in situ is recommended, rather than radical tumor removal with resection of the segment of the N. facialis involved. PMID- 16479389 TI - Characterization and application of polyclonal antibodies that specifically recognize JC virus large T antigen. AB - Polyomavirus JC virus (JCV) is the causative agent of progressive multifocal leukoencephalopathy, a fatal demyelinating disease of the central nervous system. Similar to other polyomaviruses, such as simian vacuolating virus 40 (SV40) and BK virus (BKV), JCV is also associated with human tumours. The Polyomavirus early protein large T antigen (TAg) plays a crucial role in tumour pathogenesis. An antibody to SV40 TAg (PAb416), which cross-reacts with TAgs of both JCV and BKV, has been used widely for the detection of JCV and BKV TAgs. As a consequence, it is difficult to discriminate between the TAgs of SV40, BKV and JCV by immunohistochemical analyses. In the present study, we generated JCV TAg-specific polyclonal antibodies (JCT629 and JCT652) by immunization of New Zealand white rabbits with synthetic peptides reproducing the JCV TAg carboxyl-terminal region as immunogens. Immunoblotting analyses indicated that the new antibodies bind specifically to JCV TAg, and not to those of SV40 or BKV. We also demonstrated that these antibodies can be used for immunoprecipitation, immunocytochemical analyses and immunohistochemical staining of routinely processed specimens. In conclusion, the newly generated JCV-specific TAg antibodies may be useful both in the investigation of the pathophysiological function of JCV TAg and in discriminating between Polyomavirus-related clinical samples. PMID- 16479390 TI - Assessing the efficiency of extracorporeal shockwave lithotripsy for stones in renal units with impaired function: a prospective controlled study. AB - The objective was to determine the efficiency of extracorporeal shockwave lithotripsy (ESWL) in clearing stones from renal units with impaired function. Thirty-five patients with poorly functioning kidneys determined by intravenous urogram and 99mtechnetium diethylene triamine pentacetic acid renal dynamic scan underwent ESWL. Stone clearance was assessed at 3 months and compared with that in normally functioning kidneys. The study group was divided into two subgroups. Those with split glomerular filtration rate (GFR) of the concerned kidney between 10 and 20 ml/min were in group 1. Group 2 consisted of patients with split GFR between 20 and 30 ml/min. A control group (group 3) was formed from patients with urolithiasis and normally functioning kidneys. The overall retreatment rate was 84.4%. The overall stone clearance rate in the study group was 34.2% while it was 57.7% in the control group. The stone clearance rate in group 2 was 40%. The difference in stone clearance rate between the study and control groups was statistically significant (P=0.023) but that between group 2 and the control group was not (P=0.159). The incidence of steinstrasse between the study group 2 and control group was not statistically significant (P=0.408). The clearance rate for ureteral stones was comparable in all the three groups. The stone-free rate and rate of steinstrasse for renal stones in kidneys with moderately impaired function were comparable to normally functioning kidneys. However, kidneys with severely impaired function had poor results. The clearance rate for ureteral stones was not influenced by the impairment of renal function. PMID- 16479388 TI - Long-term outcomes of the surgical management of renal cell carcinoma. AB - It has been 35 years since the radical nephrectomy was standardized by the work of Robson et al. (J Urol 101:297-301, 1969). Despite being based on a retrospective review of only 88 cases operated upon over a span of 15 years, this publication was an important milestone in the attempt to create uniformity in the staging of Renal cell carcinoma (RCC), and the measurement of surgical outcomes for RCC. Although this manuscript forms the basis for our contemporary measurement of the long-term results of RCC surgery and set the standard to which the entire subsequent literature was compared, contemporary research subsequently has questioned many of Robson's conclusions regarding RCC. In Robson's era, the majority of patients presented with large, symptomatic tumors, pre-operative staging was imprecise, and many patients had locally advanced disease at the time of surgery: of the 88 patients in Robson's series, 75% were managed through a thoracoabdominal incision. Since that time, advances in renal imaging and clinical staging have led to the increased detection of incidental, lower stage, organ-confined tumors more amendable to expanded surgical options. Surgical techniques have evolved and technological advances have made possible new methods of managing renal tumors in situ that have emphasized a transition from radical to less extirpative approaches. In addition, understanding of the basic biology and genetics of kidney cancer has led to improved prognostication and the development of effective immunotherapies for advanced disease. The current concepts and long-term outcomes of the surgical management of RCC will be reviewed to help elucidate some of these changes, from the evolution of open to laparoscopic to percutaneous, from radical to partial to ablative approaches. PMID- 16479391 TI - Continuous infusion of oxalate by minipumps induces calcium oxalate nephrocalcinosis. AB - It is hypothesized that oxalate plays an active role in calcium oxalate (CaOx) nephrocalcinosis and oxalate driven nephrolithiasis by interacting with the kidney. We developed an adjustable, nonprecursor, continuous infusion model of hyperoxaluria and CaOx nephrocalcinosis to investigate this hypothesis. Minipumps containing PBS or KOx (60-360 micromol/day; n = 5-7/dose) were implanted subcutaneously in male Sprague-Dawley rats on D0 and D6. Rats were killed on D13. Oxalate excretion and CaOx crystalluria were monitored by 20+4 h urine collections. Localization and content of intrarenal crystals were determined on frozen sections using polarization and microFTIR. Oxalate excretion was significantly elevated in all KOx rats (P < or = 0.005). CaOx crystalluria was most persistent in the 240-360 micromol/day KOx rats, but even 60 micromol/day KOx rats showed sporadic crystalluria. One hundred percent of KOx rats had CaOx nephrocalcinosis as confirmed by microFTIR. Most crystals were localized to the lumens of the corticomedullary collecting ducts. A few crystals are localized just under the papillar urothelium. The minipump model is the first model of hyperoxaluria to provide continuous infusion of oxalate. It permits control of the levels of hyperoxaluria, crystalluria and CaOx nephrocalcinosis. The level of sustained hyperoxaluria and CaOx nephrocalcinosis induced by treatment with 360 micromol/day KOx for 13D models the conditions frequently observed in jejunoileal bypass patients. Adjustments in the length of treatment and level of hyperoxaluria may allow this model to also be used to study the oxalate driven CaOx-nephrolithiasis common in patients with hyperoxaluria due to other causes. PMID- 16479392 TI - [Differential diagnosis of "sterile" phlegmonous hand infections]. AB - INTRODUCTION: Bacterial infections represent a large proportion of emergencies in hand surgery. In some cases, pyoderma gangrenosum and mycobacterial infection may present with the same symptoms of swelling, pain, and purulent secretion. In these cases, operative treatment would be harmful. Therefore two cases-pyoderma gangrenosum and tuberculosis-are presented here in relation to common bacterial hand infection. METHODS: Using two case reports of diseases that only rarely affect the hands, their relevance to differential diagnosis is shown with reference to the literature. RESULTS: In both cases, we found clinical symptoms of bacterial hand infection with negative bacterial smear tests. After several debridements, pyoderma gangrenosum of the dorsum of the hand was diagnosed in one patient after pyodermiform lesions at the thigh and the nasal septum were detected and pre-existing colitis ulcerosa was taken into consideration. Corticoid therapy induced complete remission. The second patient with similar clinical symptoms had been operated on at another hospital several times before being transferred to our institution. The presumptive diagnosis of pyoderma gangrenosum was made, and under treatment with prednisone the symptoms quickly improved. After 2 weeks, the wound conditions and the patient's condition rapidly worsened. Following amputation at the upper arm level, the patient died of septic multiple organ failure. Autopsy studies revealed tuberculous sepsis originating from the hand. DISCUSSION: Patient history should be evaluated carefully because of its value to correct diagnosis. In case of negative smear tests, especially from immunocompromised, elderly patients and in patients with a history of pulmonary tuberculosis, Ziehl-Neelsen staining should be obtained. In case of multilocular affection or pre-existing chronic inflammatory bowel disease, the presumptive diagnosis of pyoderma gangrenosum can be confirmed by biopsies from the lesions margin. In both cases, unnecessary traumatizing operations could thus be avoided and treatment optimized. PMID- 16479394 TI - Isolated dislocation of the scaphoid. AB - Isolated dislocation of the carpal scaphoid is an extremely rare injury. The authors report herein a case of a 28-year-old man managed with open reduction and Kirschner wire fixation. The aim of this study is to comprehensively present this unusual injury along with its treatment and to attempt to merge the available experience in the literature in a suggested algorithm that will guide the surgeon confronted with this rare problem to treat it promptly and effectively. PMID- 16479393 TI - [Oblique osteotomy for post-traumatic fibula incongruity]. AB - BACKGROUND: Injuries of the ankle joint are common and, considering that this joint is highly mechanically stressed, it seems obvious that therapy requires restoring the joint to a physiological, anatomically functional unit. In case of post-traumatic shortening, rotation and talar tilt arthritis may result because of the changed biomechanics. Before this occurs, typical clinical symptoms are recurrent swelling, pain, and limited range of motion, together resulting in reduced mobility and quality of life. With prompt indication and revision lengthening the lateral ankle and thus restoring joint congruity, patient complaints are significantly reduced and the development of advanced arthritis can be avoided. PATIENTS AND METHODS: Within a period of 7 years, 16 patients with post-traumatic shortening of the fibula were treated with oblique osteotomy and lengthening of the fibula at our department. The mean time between trauma and reconstructive surgery was 10 months, and the median follow-up period to clinic and radiological examination was 3.7 years. RESULTS AND CONCLUSION: Comparison of results before and after correction showed significant subjective improvements, and the development or continuation of post-traumatic arthritis could be hindered, of critical importance to which is prompt indication for operative joint correction, because clinical symptoms of advanced osteoarthritis of the ankle joint cannot be expected to improve. PMID- 16479395 TI - Management of a nonunion of the distal femur in osteoporotic bone with the internal fixation system LISS (less invasive stabilization system). AB - We present a case illustrating the successful use of the internal fixation LISS in an osteoporotic nonunion of the distal femur, where classic osteosynthesis has failed. The LISS plate with its angular stability offered the possibility to achieve excellent purchase in the severely porotic and partially destroyed bone. In combination with the use of an autologeous bone graft laterally and a strut cortical autograft medially, a mechanical support and an osteoinductive stimulus was provided and the extremity could be saved by this procedure. PMID- 16479396 TI - Posterior atlantoaxial three-point fixation: comparison of intraoperative performance between open and percutaneous techniques. AB - INTRODUCTION: Atlantoaxial instabilities, which require surgical fixation follow a variety of clinical disorders. Different surgical procedures are used for stabilization of the atlantoaxial complex, mainly posterior wiring techniques and transarticular screw fixation. Nowadays, often a combination of transarticular screws and a posterior one-point fixation is used to achieve a three-point fixation, with superior biomechanical stability and good clinical results. Different modifications were developed to improve this technique. In 1995, a percutaneous approach for atlantoaxial stabilization was introduced. In clinical studies, the technique showed a tendency towards better outcome. Beside the outcome, the intraoperative performance is of special interest for minimal invasive approaches. We therefore compared the operation time, screw angulation and blood loss, between the open and percutaneous posterior atlantoaxial techniques. MATERIALS AND METHODS: Two groups, each consisting of 17 patients, with either open (group 1) or percutaneous (group 2) atlantoxial stabilization, were compared. The operation time was retrospectively acquired from the patient's charts. The data for blood loss was provided by our anaesthesiological department, separated for intraoperative, postoperative and total blood loss. Screw angulation was measured on the postoperative x-ray by an orthopaedic surgeon. RESULTS: The percutaneous group showed an average intraoperative blood loss of 239.7 ml, compared to 929.4 ml for the open group (p< or =0.001). The analogue values for the postoperative blood loss were 142.9 ml and 379.4 ml for group 2 and group 1, respectively (p=0.008). Consecutively, the total blood loss showed also a statistically significant difference (p< or =0.001). The operation time was significantly different (p< or =0.001), with average values of 175.3 min (group 1) and 110.6 min (group 2). Screw angulation showed a trend towards a steeper angulation in the percutaneous group with an average angle of 56.8 degrees , compared to 53.9 degrees (group 1), although this was not statistically significant (p=0.053). CONCLUSION: The percutaneous technique for atlantoaxial stabilization with a three-point fixation has clear intraoperative benefits, with shorter operation time and reduced blood loss. A trend towards steeper screw angulation was found and shows at least equal feasibility for transarticular screw placement with the percutaneous technique, compared to the standard open approach. PMID- 16479397 TI - Object grasping using the minimum variance model. AB - Reaching-to-grasp has generally been classified as the coordination of two separate visuomotor processes: transporting the hand to the target object and performing the grip. An alternative view has recently been formed that grasping can be explained as pointing movements performed by the digits of the hand to target positions on the object. We have previously implemented the minimum variance model of human movement as an optimal control scheme suitable for control of a robot arm reaching to a target. Here, we extend that scheme to perform grasping movements with a hand and arm model. Since the minimum variance model requires that signal-dependent noise be present on the motor commands to the actuators of the movement, our approach is to plan the reach and the grasp separately, in line with the classical view, but using the same computational model for pointing, in line with the alternative view. We show that our model successfully captures some of the key characteristics of human grasping movements, including the observations that maximum grip size increases with object size (with a slope of approximately 0.8) and that this maximum grip occurs at 60-80% of the movement time. We then use our model to analyse contributions to the digit end-point variance from the two components of the grasp (the transport and the grip). We also briefly discuss further areas of investigation that are prompted by our model. PMID- 16479398 TI - Encoding of time-varying stimuli in populations of cultured neurons. AB - We wondered whether random populations of dissociated cultured cortical neurons, despite of their lack of structure and/or regional specialization, are capable of modulating their neural activity as the effect of a time-varying stimulation - a simulated 'sensory' afference. More specifically, we used localized low frequency, non-periodic trains of stimuli to simulate sensory afferences, and asked how much information about the original trains of stimuli could be extracted from the neural activity recorded at the different sites. Furthermore, motivated by the results of studies performed both in vivo and in vitro on different preparations, which suggested that isolated spikes and bursts may play different roles in coding time-varying signals, we explored the amount of such 'sensory' information that could be associated to these different firing modes. Finally, we asked whether and how such 'sensory' information is transferred from the sites of stimulation (i.e., the 'sensory' areas), to the other regions of the neural populations. To do this we applied stimulus reconstruction techniques and information theoretic concepts that are typically used to investigate neural coding in sensory systems.Our main results are that (1) slow variations of the rate of stimulation are coded into isolated spikes and in the time of occurrence of bursts (but not in the bursts' temporal structure); (2) increasing the rate of stimulation has the effect of increasing the proportion of isolated spikes in the average evoked response and their importance in coding for the stimuli; and, (3) the ability to recover the time course of the pattern of stimulation is strongly related to the degree of functional connectivity between stimulation and recording sites. These observations parallel similar findings in intact nervous systems regarding the complementary roles of bursts and tonic spikes in encoding sensory information.Our results also have interesting implications in the field of neuro-robotic interfaces. In fact, the ability of populations of neurons to code information is a prerequisite for obtaining hybrid systems, in which neuronal populations are used to control external devices. PMID- 16479399 TI - Early embryonic development of the central nervous system in the Australian crayfish and the Marbled crayfish (Marmorkrebs). AB - This study sets out to provide a systematic analysis of the development of the primordial central nervous system (CNS) in embryos of two decapod crustaceans, the Australian crayfish Cherax destructor (Malacostraca, Decapoda, Astacida) and the parthenogenetic Marbled crayfish (Marmorkrebs, Malacostraca, Decapoda, Astacida) by histochemical labelling with phalloidin, a general marker for actin. One goal of our study was to examine the neurogenesis in these two organisms with a higher temporal resolution than previous studies did. The second goal was to explore if there are any developmental differences between the parthenogenetic Marmorkrebs and the sexually reproducing Australian crayfish. We found that in the embryos of both species the sequence of neurogenetic events and the architecture of the embryonic CNS are identical. The naupliar neuromeres proto-, deuto-, tritocerebrum, and the mandibular neuromeres emerge simultaneously. After this "naupliar brain" has formed, there is a certain time lag before the maxilla one primordium develops and before the more caudal neuromeres follow sequentially in the characteristic anterior-posterior gradient. Because the malacostracan egg nauplius represents a re-capitulation of a conserved ancestral information, which is expressed during development, we speculate that the naupliar brain also conserves an ancestral piece of information on how the brain architecture of an early crustacean or even arthropod ancestor may have looked like. Furthermore, we compare the architecture of the embryonic crayfish CNS to that of the brain and thoracic neuromeres in insects and discuss the similarities and differences that we found against an evolutionary background. PMID- 16479401 TI - Cloning and functional characterization of a fatty acid synthase component FAS2 gene from Saccharomyces kluyveri. AB - A gene coding the alpha subunit of fatty acid synthase (FAS2) was isolated from the budding yeast Saccharomyces kluyveri. Nucleotide sequence analysis indicated that this gene, termed Sk-FAS2, coded a protein having an amino acid sequence 83% identical to the FAS2 protein of S. cerevisiae (Sc-FAS2). The Sk-FAS2 gene was able to functionally complement an S. cerevisiae fas2 disruptant. This Sk-FAS2 expressing strain was found to produce larger amounts of C18 than C16, in contrast to the Sc-FAS2-expressing fas2 strain. In addition, fusion genes of Sk FAS2 and Sc-FAS2 were transformed into a fas2-disrupted strain of S. cerevisiae, and fatty acid analysis of these transformants suggested that the region containing the acyl carrier protein and beta-ketoacyl reductase domains of yeast FAS2 protein play an important role in determining carbon chain length of fatty acids. PMID- 16479403 TI - Cafe-au-lait spots in neurofibromatosis type 1 and in healthy control individuals: hyperpigmentation of a different kind? AB - Solitary cafe-au-lait spots are quite common in the general population but multiple cafe-au-lait macules (CALM) are often indicative of an underlying genetic disorder. The frequency of having more than five CALM is rare in normal individuals and is therefore considered as a cut-off for the diagnosis of neurofibromatosis type 1 (NF1). The etiopathogenesis of these macules is still very obscure. In this study we compared epidermal melanocyte and dermal mast cell numbers between four groups: control normal and control CALM skin, and NF1 normal and NF1 CALM skin and elaborated a possible role for stem cell factor (SCF) in CALM formation. The groups were analyzed by immunohistochemistry for numerical analysis of the melanocyte and mast cell population and by ELISA, western blot analysis and real-time quantitative PCR for further determination of the role of SCF. We found a significant increase in melanocyte density in NF1 CALM skin compared with the isolated CALM in control individuals. However, both groups displayed a similar increase in mast cell density. In addition, we found increased levels of soluble SCF in NF1 CALM and in NF1 normal fibroblast supernatant. We conclude that SCF is an important cytokine in NF1 skin, but that additional (growth) factors and/or genetic mechanisms are needed to induce NF1 specific CALM hyperpigmentation. PMID- 16479402 TI - Characterization of the arom gene in Rhizoctonia solani, and transcription patterns under stable and induced hypovirulence conditions. AB - The quinate pathway is induced by quinate in the wild-type virulent Rhizoctonia solani isolate Rhs 1AP but is constitutive in the hypovirulent, M2 dsRNA containing isolate Rhs 1A1. Constitutive expression of the quinate pathway results in downregulation of the shikimate pathway, which includes the pentafunctional arom gene in Rhs 1A1. The arom gene has 5,323 bp including five introns as opposed to a single intron found in arom in ascomycetes. A 199-bp upstream sequence has a GC box, no TATAA box, but two GTATTAGA repeats. The largest arom transcript is 5,108 nucleotides long, excluding the poly(A) tail. It contains an open reading frame of 4,857 bases, coding for a putative 1,618 residue pentafunctional AROM protein. A Kozak sequence (GCGCCATGG) is present between +127 and +135. The 5'-end of the arom mRNA includes two nucleotides (UA) that are not found in the genomic sequence, and are probably added post transcriptionally. Size and sequence heterogeneity were observed at both 5'- and 3'-end of the mRNA. Northern blot and suppression subtractive hybridization analyses showed that presence of a low amount of quinate, inducer of the quinate pathway, resulted in increased levels of arom mRNA, consistent with the compensation effect observed in ascomycetes. PMID- 16479404 TI - Preoperative angiography with embolization and radiofrequency ablation as novel adjuncts to safe surgical resection of a large, vascular sacrococcygeal teratoma. AB - Sacrococcygeal teratomas (SCTs) can present a challenging problem and can be associated with significant perinatal morbidity and mortality. A female child was born at 36 weeks' gestation with a large, vascular Type 1 SCT originally identified by prenatal ultrasound. A CT scan showed two large feeding vessels arising from both internal iliac arteries that were successfully embolized during angiography. A radiofrequency probe was then used to ablate a zone between normal tissue and the tumor. The SCT was subsequently surgically excised with minimal blood loss. This case is presented to illustrate two useful and previously unreported postnatal adjuncts to the surgical treatment of massive, hypervascular sacrococcygeal tumors. PMID- 16479405 TI - Antioxidant activity in Spalax ehrenbergi: a possible adaptation to underground stress. AB - The blind subterranean mole rat Spalax ehrenbergi superspecies has evolved adaptive strategies to cope with underground stress. Hypoxia is known to stimulate reactive oxygen species generation; however, mechanisms by which Spalax counteracts oxidative damage have not been investigated before. We studied in Spalax the oxidative status of the Harderian gland (HG), an organ which is particularly vulnerable to oxidative stress in many rodents. With regard to the sexual dimorphism found in this gland, differences between males and females were determined and compared to the surface-dwelling Syrian hamster. Our results show, for the first time, that Spalax exhibits remarkably low biomolecular damage, which implies the existence of physiological strategies to avoid oxidative damage under fluctuating O(2) and CO(2) levels existing in the mole rat's subterranean niche. Correspondingly, main antioxidant enzymes, such as superoxide dismutase (SOD), catalase, and glutathione reductase (GR), exhibited high activities in both genders; in particular, remarkably high levels were measured in SOD. SOD and GR activities showed statistically significant differences between sexes. Melatonin, an important circadian agent is also a very important antioxidant molecule and is synthesized in the Harderian glands (HGs) of Spalax. Therefore, the possible interaction between antioxidant enzymes and melatonin is suggested. PMID- 16479407 TI - Patient safety: lessons learned. AB - The traditional approach to patient safety in health care has ranged from reticence to outward denial of serious flaws. This undermines the otherwise remarkable advances in technology and information that have characterized the specialty of medical practice. In addition, lessons learned in industries outside health care, such as in aviation, provide opportunities for improvements that successfully reduce mishaps and errors while maintaining a standard of excellence. This is precisely the call in medicine prompted by the 1999 Institute of Medicine report "To Err Is Human: Building a Safer Health System." However, to effect these changes, key components of a successful safety system must include: (1) communication, (2) a shift from a posture of reliance on human infallibility (hence "shame and blame") to checklists that recognize the contribution of the system and account for human limitations, and (3) a cultivation of non-punitive open and/or de-identified/anonymous reporting of safety concerns, including close calls, in addition to adverse events. PMID- 16479408 TI - Imaging in cancer therapy and drug development. PMID- 16479406 TI - Evidence of an oscillating peripheral clock in an equine fibroblast cell line and adipose tissue but not in peripheral blood. AB - The master mammalian pacemaker in the brain controls numerous diverse physiological and behavioral processes throughout the organism. Timing information is continually transmitted from the master clock to peripheral organs to synchronize rhythmic daily oscillations of clock gene transcripts and control local physiology. To investigate the presence of peripheral clocks in the horse, quantitative real-time RT-PCR assays were designed to detect levels of equine clock genes. Expression profiles for Per2, Bmal1 and Cry1 were first determined in a synchronized equine cell line. Subsequently, expression in equine whole blood and adipose tissue was assessed. Robust circadian oscillations of Per2, Bmal1 and Cry1 were observed in vitro. A synchronized molecular clock was also demonstrated in equine adipose tissue although oscillation of Bmal1 was less robust than that of Per2 and Cry1. In contrast to previous studies in humans and rats however, there was no evidence of synchronized clock gene expression in equine peripheral blood. These studies suggest that synchronous control of clock gene oscillation in equine peripheral blood is not as tightly regulated as in other species and may reflect the influence of different evolutionary challenges modifying the function of a peripheral clock. PMID- 16479411 TI - Spatial distribution of primates in a mosaic of colonizing and old growth forest at Ngogo, Kibale National Park, Uganda. AB - Primate censuses were conducted in a mosaic of colonizing (two locations) and old growth forests using line transect methods at the Ngogo study site, Kibale National Park, Uganda. Black and white colobus monkeys (Colobus guereza) were encountered more frequently in the colonizing forests than in the old growth forest, while chimpanzees (Pan troglodytes) were encountered more frequently in the old growth forest than in colonizing forests. Although not significant, results suggest that blue monkeys (Cercopithecus mitis) frequented colonizing forests more often than old growth forest. The encounter rates of mangabey (Lophocebus albigena), and redtail (Cercopithecus ascanius) groups were ambiguous with their density being higher in some colonizing forests but not others as compared to old-growth forest. No significant differences were detected for baboons (Papio anubis), L'hoest's (Cercopithecus lhoesti), and red colobus monkeys (Piliocolobus tephroscales). The conversion of forests to farmland is one of the major problems encountered in primate conservation. This study shows that secondary forests replacing anthropogenic grasslands have the potential of supporting some primate species such as black and white colobus, redtail monkeys, and possibly blue monkeys. Therefore, such areas should not be given up but should be conserved for the benefit of primates that can survive in secondary forests; as the forests mature further, primate species that are adapted to old growth forest will colonize the area provided there is a nearby source. PMID- 16479410 TI - Intraarticular migration of a broken biodegradable interference screw after anterior cruciate ligament reconstruction. AB - Poly-L: -lactic acid biodegradable screws have been used effectively for graft fixation in anterior cruciate ligament (ACL) reconstruction. The overall complication rate associated with the use of this implant is low, although some authors reported complications, such as osteolysis and aseptic effusion of the knee joint. We report a case of a 29-year-old female patient with a failure of a biodegradable interference screw at 22 months after ACL reconstruction using bone patellar tendon-bone graft. In this illustrated case, the screw broke and migrated into the knee joint. In addition, we performed a detailed review of the medical literature from 1990-2005 to identify possible causes of biodegradable screw failures. We identified six published cases of bioabsorbable interference screw failure with migration into the knee joint. Several authors have reported small diameter of the screw, poor bone quality, bone resorption, and screw divergence as potential causes for intraarticular migration of metallic interference screws. With regard to bioscrews, no specific risk factors for screw breakage and intraarticular migration have been reported. ACL reconstruction with the use of bioabsorbable interference screws for fixation is considered to be reliable. However, we need to be aware of potential problems associated with the use of this implant. Early recognition of bioscrew failure may prevent associated morbidities, such as subsequent cartilage damage. PMID- 16479409 TI - [Pharmacodynamics of two different propofol formulations]. AB - BACKGROUND: Propofol is nowadays available in various lipid formulations. We compared two different propofol formulations with respect to pharmacodynamics, using the EEG and clinical signs. MATERIALS AND METHODS: Ten volunteers received Diprivan 1% and Propofol 1% MCT Fresenius as a computer controlled infusion with increasing propofol target concentrations. A sigmoid E(max) model with effect compartment was estimated for the median frequency of the EEG power spectrum, based on measured arterial propofol plasma concentrations. Clinical pharmacodynamics were assessed by reaction on acoustic stimuli, eyelid reflex and corneal reflex. RESULTS: The drugs did not differ in pharmacodynamics with respect to EEG (EC(50) 2.1+/-0.6 for Diprivan and 2.1+/-0.5 microg/ml for Propofol Fresenius) and clinical signs. The pharmacodynamic model was characterized by a steep concentration effect relationship and a distinct hysteresis between propofol plasma concentration and effect (k(e0) 0.12+/-0.04 and 0.12+/-0.5 min(-1)). CONCLUSIONS: The investigated lipid formulations have no influence on the pharmacodynamics of propofol. PMID- 16479412 TI - HIV education needs among Sudanese immigrants and refugees in the Midwestern United States. AB - Migrant populations may be particularly vulnerable to infectious diseases and often need special considerations in terms of health care. In particular, migrant populations can be at high risk for HIV infection, in part due to lack of education about disease acquisition. This study evaluated knowledge, attitudes, and beliefs about HIV/AIDS as well as risk behavior in the Sudanese immigrant and refugee population of Nebraska (N = 47). The results demonstrated that a significant proportion of individuals from this population are poorly educated about HIV infection, exhibit attitudes and beliefs that may increase their risk for disease acquisition, and create barriers to HIV prevention and care, and engage in high-risk sexual behaviors. Appropriate educational materials are lacking, and there is a pressing need for improved access to culturally appropriate HIV education for this vulnerable population. PMID- 16479413 TI - Severity of child sexual abuse, post- traumatic stress and risky sexual behaviors among HIV-positive women. AB - Child sexual abuse (CSA) has been shown to enhance risk for HIV infection and other adverse outcomes. However, most studies examine the effects of a single incident of CSA rather than the full burden of abuse over the life span in predicting these adverse outcomes. A multi-dimensional approach was used in this study to examine the severity of abuse as a predictor of post-traumatic stress, depression, sexual symptoms, and risky sexual behaviors in a multi-ethnic sample of 147 HIV-positive women. Multivariate models indicated that experiencing both intrafamilial and extrafamilial CSA, adult sexual abuse (ASA) and Latina ethnicity predicted PTSD symptoms. ASA also predicted sexual trauma symptoms. Also, CSA and adult re-victimization contributed independently to risk for PTSD and sexual trauma symptoms, but not for risky sexual behaviors. The results support the need for interventions for HIV-positive women that address the full burden of abuse experienced and its sequelae. PMID- 16479414 TI - Predictors of sexually transmitted infection testing among sexually active homeless youth. AB - We examined the association between sexual risk behaviors and sexually transmitted infection (STI) testing in a sample of homeless youth. Of 261 youth interviewed, 50% had been sexually active in the past 3 months. Gender variation in sexual behaviors and risk were found. Boys were more likely than girls to engage in anal sex (46% vs. 15%), to have 3 or more sexual partners (46% vs. 17%) and to engage in anonymous sex (38% vs. 21%). Girls were less likely to use condoms consistently and more likely to engage in sex with a partner suspected of having an STI (20% vs. 4%). In the past 3 months, the STI testing rates were similar for boys and girls (46%). However, girls were more likely to have positive STI results (46% vs. 9%). In a multivariate logistic regression analysis, the only variable that was an independent predictor of STI testing was having either gotten someone or having become pregnant in the past 3 months. High risk sexual behaviors did not predict STI testing in our sample. Outreach programs are needed that target sexually active homeless youth for early STI testing and treatment. PMID- 16479415 TI - Preparation, premedication, and surveillance. PMID- 16479416 TI - Pancreatic ERCP. AB - Like biliary endoscopy, pancreatic endoscopic retrograde pancreatography (ERCP) has evolved from being a diagnostic procedure to a therapeutic one. The advent of magnetic resonance cholangiopancreatography and endoscopic ultrasonography have limited the need for anatomic definition and transpapillary tissue sampling, respectively. This review is therefore limited to studies published during the past year concerned either with efforts to limit the incidence of post-ERCP pancreatitis or with critical reviews of therapeutic pancreatography, including the treatment of strictures, stones, and ductal disruption. PMID- 16479417 TI - Gastrointestinal bleeding: from overt to obscure. PMID- 16479418 TI - Latest developments in the endoscopic management of gastroesophageal reflux disease and Barrett's esophagus: an overview of the year's literature. AB - The development of endoscopic techniques for the treatment of gastroesophageal reflux disease has come to an abrupt halt after some of the most widely disseminated and best evaluated techniques were withdrawn from the market. This underlines the importance of conducting high-quality endoscopic research before techniques are adopted in routine practice. The endoscopic surveillance of patients with Barrett's esophagus needs to be improved in order to be cost effective. Alternative techniques for tissue sampling are being explored (e. g., brush cytology, imaging cytometry), as well as the use of molecular markers, to select patients who are at risk and to improve compliance by patients. New endoscopic imaging techniques (such as autofluorescence endoscopy) may improve the detection of early neoplasia in those Barrett's patients who are at risk for malignant degeneration. Subsequent endoscopic treatment should primarily consist of an endoscopic resection technique with restricted use of ablative therapies. Future studies should focus on the development of endoscopic resection techniques that allow en-bloc resection of Barrett's lesions, stepwise complete resection of the whole Barrett's segment, and/or easier and safer tissue ablation. Finally, histopathological studies are needed in order to detect risk factors for an adverse outcome after endoscopic treatment for early neoplasia in Barrett's patients. PMID- 16479419 TI - Upper gastrointestinal tumors. PMID- 16479420 TI - Minimally invasive surgery, 2004-2005. PMID- 16479421 TI - The Aer-O-Scope: proof of the concept of a pneumatic, skill-independent, self propelling, self-navigating colonoscope in a pig model. AB - BACKGROUND AND STUDY AIMS: Considerable training is needed to enable endoscopists to use the currently available commercial colonoscopes and sigmoidoscopes effectively and safely. The aim of this study was to evaluate the safety and efficacy of the propulsion mechanism incorporated into the Aer-O-Scope (GI View Ltd., Ramat Gan, Israel)--a novel self-propelled, self-navigating colonic endoscope for diagnostic purposes. MATERIALS AND METHODS: Twenty young female pigs underwent complete bowel preparation followed by a sedated examination using the new device. Ten pigs underwent two consecutive procedures, with a wash-out period of 7 days between each procedure. The total number of procedures was 30. Two prototypes of the Aer-O-Scope, with different cable lengths and vehicle balloon sizes (n = 20 and n = 10 for prototypes I and II, respectively) were used. Each examination was followed by a standard colonoscopy for safety evaluation. The insertion length of the Aer-O-Scope was determined by fluoroscopy images. RESULTS: The colon was adequately clean in 25 procedures. Maximum insertion was achieved in 21 procedures (84%)--80% with prototype I (n = 15) and 90% with prototype II (n = 10). The time to maximum insertion averaged 8.9 +/- 4.4 min (10 +/- 4.6 and 6.6 +/- 2.9 min for prototypes I and II; P < 0.05), and the withdrawal time averaged 3.4 +/- 2.1 and 4.2 +/- 3.4 min, respectively. The driving pressures for prototypes I and II averaged 46.3 and 34.5 mbar, respectively. The follow-up conventional colonoscopy identified no mucosal tears or perforations. Minor mucosal petechiae were noted in 43% of the cases. No adverse events were noted up to 7 days after the procedure. CONCLUSIONS: The propulsion mechanism used in this novel self-propelled, self-navigating colonoscope was effective and safe in pigs. PMID- 16479423 TI - Clinical impact of conventional endosonography and endoscopic ultrasound-guided fine-needle aspiration in the assessment of patients with Barrett's esophagus and high-grade dysplasia or intramucosal carcinoma who have been referred for endoscopic ablation therapy. AB - BACKGROUND AND STUDY AIMS: Endoscopic mucosal resection and photodynamic therapy are exciting, minimally invasive curative techniques that represent an alternative to surgery in patients with Barrett's esophagus and high-grade dysplasia or intramucosal adenocarcinoma. However, there is lack of uniformity regarding which staging method should be used prior to therapy, and some investigators even question whether staging is required prior to ablation. We report our experience with a protocol of conventional endoscopic ultrasound staging prior to endoscopic therapy. PATIENTS AND METHODS: A total of 25 consecutive patients with a diagnosis of high-grade dysplasia or intramucosal adenocarcinoma in Barrett's esophagus who had been referred to the University of Chicago for staging in preparation for endoscopic therapy between March 2002 and November 2004 were included in the study. All 25 patients underwent repeat diagnostic endoscopy and conventional endosonography with a radial echo endoscope. Any suspicious lymph nodes that were detected were sampled using endoscopic ultrasound-guided fine-needle aspiration. RESULTS: Baseline pathology in the 25 patients (mean age 70, range 49-85) revealed high-grade dysplasia in 12 patients and intramucosal carcinoma in 13 patients. Five patients were found to have submucosal invasion on conventional endosonography. Seven patients had suspicious adenopathy, six regional (N1) and one metastatic to the celiac axis (M1a). Fine-needle aspiration confirmed malignancy in five of these seven patients. Based on these results, five patients (20%) were deemed to be unsuitable candidates for endoscopic therapy. CONCLUSIONS: By detecting unsuspected malignant lymphadenopathy, conventional endosonography and endoscopic ultrasound with fine-needle aspiration dramatically changed the course of management in 20% of patients referred for endoscopic therapy of Barrett's esophagus with high-grade dysplasia or intramucosal carcinoma. Based on our results, we believe that conventional endosonography and endoscopic ultrasound with fine-needle aspiration when nodal disease is present should be performed routinely in all patients referred for endoscopic therapy in this setting. PMID- 16479422 TI - High rate of lymph-node metastasis in submucosal esophageal squamous-cell carcinomas and adenocarcinomas. AB - BACKGROUND AND STUDY AIMS: The application of endoscopic mucosectomy in early esophageal cancer is limited by the presence of lymph-node metastasis. The aim of this prospective study was to analyze the rate of lymph-node involvement relative to the depth of mucosal or submucosal tumor penetration, comparing squamous-cell carcinomas and adenocarcinomas. PATIENTS AND METHODS: A total of 60 patients with pT1 esophageal cancer--24 with squamous-cell carcinomas (SCCs) and 36 with adenocarcinomas--were treated with transthoracic en-bloc esophagectomy with two field lymphadenectomy (n = 50) or transhiatal esophageal resection (n = 10). An average of 30 lymph nodes were examined, and the following characteristics were evaluated: histology, mucosal infiltration, depth of submucosal wall infiltration in three thirds (sm1, sm2, sm3), grading, resection category, ratio of metastatic to resected lymph nodes, and locations of metastatic nodes. RESULTS: The rates of lymph-node metastasis were 0% for the 16 mucosal carcinomas and 45% for the 44 submucosal carcinomas (P < 0.01). There were no significant differences in the extent of lymph-node involvement between submucosal adenocarcinomas (41%) and submucosal SCCs (50%). Sm1 carcinomas were associated with a lower rate of lymph node metastasis (SCCs 33%, adenocarcinomas 22%) than sm3 carcinomas (SCCs 69%, adenocarcinomas 78%). Two patients (9%) with submucosal SCCs and five patients (23%) with submucosal adenocarcinomas were classified as having stage pM1 lymph. The average lymph-node ratio in patients with pN1 was 0.13 for adenocarcinomas and 0.1 for SCCs (difference not significant). In the multivariate analysis, the parameters mucosal vs. submucosal (P < 0.01) and G1/G2 vs. G3 (P < 0.05) showed a significant impact in relation to metastatic lymph nodes. CONCLUSIONS: The most important factor for predicting lymph-node metastasis in early esophageal cancer is the presence of submucosal infiltration. Early adenocarcinomas and SCCs do not differ with regard to their rate of lymphatic involvement. The rate of lymph-node metastasis increases with the depth of submucosal infiltration, but metastases can already occur in sm1 lesions. Submucosal infiltration is a contraindication for endoscopic mucosectomy. Limited surgical procedures without adequate lymphadenectomy do not appear to be appropriate in the treatment of patients with submucosal esophageal carcinomas. PMID- 16479424 TI - Surveillance in Barrett's oesophagus: will a strategy focused on a high-risk group reduce mortality from oesophageal adenocarcinoma? AB - BACKGROUND AND STUDY AIMS: The incidence of oesophageal adenocarcinoma has increased significantly in recent years. While surveillance of people with Barrett's oesophagus, its usual precursor, has been advocated in order to detect dysplasia and early cancer in those considered to be at greatest risk, the impact of such a strategy on survival from oesophageal adenocarcinoma is unclear. This study aimed to determine the effect of surveillance on mortality from oesophageal adenocarcinoma in a group of patients considered to be at high risk of developing Barrett's oesophagus and adenocarcinoma. PATIENTS AND METHODS: After performing a Medline search of the literature published between 1985 and 2004 for studies on gastro-oesophageal reflux disease, Barrett's oesophagus and adenocarcinoma, we examined the impact of surveillance on mortality from oesophageal adenocarcinoma in a hypothetical sample of 100 high-risk patients (men aged over 50 with Barrett's oesophagus but without high-grade dysplasia at entry). RESULTS: Four patients in this high-risk group developed adenocarcinoma during surveillance, with survival rates of 78.9% (95%CI 64.9%-88.5%) at 2 years and 78.6% (95%CI 62.8%-89.2%) at 5 years. Meanwhile, between 515 and 2060 patients with Barrett's oesophagus were not detected or surveyed by this strategy and between 16 and 61 of these developed adenocarcinoma, with much lower survival rates of 37.1% (95%CI 25.4%-50.3%) at 2 years and 16.7% (95%CI 9%-28.3%) at 5 years. Although surveillance in the high-risk group resulted in the long-term survival of three patients who would not otherwise have survived, this gain was dramatically offset by the 13 to 51 patients, excluded from surveillance by this strategy, who died from oesophageal adenocarcinoma. CONCLUSIONS: A surveillance programme based on current concepts of risk cannot have an impact on mortality from oesophageal adenocarcinoma. To be effective, it will be necessary for surveillance programmes to utilise more precise methods for the identification of those who are most at risk of progression to adenocarcinoma. PMID- 16479425 TI - Efficacy and safety of endoscopic submucosal dissection for tumors of the esophagogastric junction. AB - BACKGROUND AND STUDY AIMS: The esophagogastric junction (EGJ) has been considered a difficult location for endoscopic treatment of tumors, due to its narrow lumen and sharp angle. Endoscopic submucosal dissection (ESD) is a method of endoscopic resection, capable of removing large tumors in an en bloc fashion. The aim of this study was to evaluate the efficacy and safety of ESD for EGJ tumors. PATIENTS AND METHODS: For 30 lesions of EGJ tumors treated by ESD, the size of the lesions and resected specimens, the en bloc resection rate, complications, and local recurrence were assessed. RESULTS: The average maximum diameters of the lesions and resected specimens were 22.4 mm and 40.6 mm respectively. The complete en bloc resection (R0) rate was 97% (29/30). Histological evaluation of the resected specimens revealed five cases of angiolymphatic invasion and five cases of submucosal invasion deeper than 500 microm. Perforation occurred in one case but was safely managed by rotatable clips and administration of antibiotics for 3 days. Local recurrence was not observed in any patient during follow-up (mean 14.6 months, range 6-31 months). CONCLUSIONS: ESD can be safely performed for EGJ tumors, with a high en bloc resection rate. For lesions with no apparent submucosal invasion findings, ESD is a curative and diagnostic treatment option that may be considered before open surgery. PMID- 16479426 TI - Endoscopic mucosal resection and the risk of lymph-node metastases: indications revisited? PMID- 16479427 TI - UEGW congress report, Copenhagen 2005: Review of endoscopic abstracts. PMID- 16479428 TI - Indocyanine green is an ideal dye for endoscopic ultrasound-guided fine-needle tattooing of pancreatic tumors. AB - Endoscopic ultrasound-guided fine-needle tattooing (EUS-FNT) is an ideal technique for preoperative marking of lesions detected on preoperative examination. Although India ink has been used for endoscopic tattooing, there have been numerous reports of complications associated with its use. This is the first report of EUS-FNT using indocyanine green (ICG) and describes its use for preoperative marking of a tumor in a 78-year-old man with multiple pancreatic tumors. There were no complications associated with the EUS-FNT procedure and it is suggested that ICG is a more suitable dye for tattooing of pancreatic lesions than India ink, being far less frequently associated with side effects. PMID- 16479429 TI - Endoscopic band ligation of bleeding dieulafoy lesions: the best therapeutic strategy. PMID- 16479430 TI - A novel "stylet" device for facilitating the advancement of a colonoscope. PMID- 16479431 TI - Successful treatment of gastroparesis following total esophagectomy using botulinum toxin. PMID- 16479432 TI - Colonoscopic treatment of gallstone ileus. PMID- 16479433 TI - Endoscopic findings and treatment of a recurrent adenocarcinoma with a submucosal tumor-like appearance in short-segment Barrett's esophagus. PMID- 16479434 TI - Endoscopic ultrasound-guided drainage of a biloma: a novel approach. PMID- 16479441 TI - Homocyst(e)ine and stroke. AB - Homocyst(e)ine elevation is associated with a two- to threefold fold increased risk of ischemic stroke. Although most commonly associated with large-artery atherosclerosis and venous thrombosis, hyperhomocysteinemia may contribute to stroke by other mechanisms as well. Levels of homocysteine are determined by genetic regulation of the enzymes involved in homocyst(e)ine metabolism and by levels of the vitamin cofactors (folate, B (6), and B (12)) associated with those reactions. Emerging evidence suggests that genetic variation within this pathway, such as the methyleneterahydrofolate reductase and cystathionine beta-synthase and nicotinamide N-methyltransferase genes, increases the risk of ischemic stroke. The introduction of grain folate fortification in 1998 has reduced homocyst(e)ine concentrations in the U.S. population. However, it is important to screen for vitamin B (12) deficiency and be cognizant that vitamin B (6) levels may be low in the elderly and in individuals with inflammatory disorders. The Vitamin Intervention in Stroke Prevention study failed to prove that high-dose supplementation with folate, B (6), and B (12) reduced the risk of recurrent stroke or myocardial infarction at 2 years; however, there is an ongoing clinical trial evaluating the potential benefit of vitamin supplementation. PMID- 16479440 TI - Optimal dietary habits for the prevention of stroke. AB - Diet may influence stroke risk via several mechanisms, but the optimal dietary habits for stroke prevention are not well established. We reviewed English language MEDLINE publications from January 1979 through November 2004 for experimental, observational, and clinical studies of dietary factors (minerals, fats, cholesterol, fish, animal protein, fiber, whole grains, carbohydrate quality, fruits and vegetables, antioxidants, B vitamins, dietary patterns) and risk of stroke or hypertension, the principal modifiable stroke risk factor. A total of 121 publications were selected based on relevance and quality of design and methods. Diets low in sodium and high in potassium lower blood pressure which will likely reduce stroke risk. Consumption of fruits and vegetables, whole grains, folate, and fatty fish are each likely to reduce stroke risk. A prudent or traditional Mediterranean dietary pattern, which incorporates these individual dietary components as well as intake of legumes and olive oil, may also prevent stroke. Evidence is limited or inconsistent regarding optimal levels of dietary magnesium, calcium, antioxidants, total fat, other fat subtypes, cholesterol, carbohydrate quality, or animal protein for stroke prevention. A diet low in sodium, high in potassium, and rich in fruits, vegetables, whole grains, cereal fiber, and fatty fish will likely reduce the incidence of stroke. Further research is needed regarding the possible effects of other major dietary factors on stroke risk. PMID- 16479442 TI - Single-gene stroke disorders. AB - Monogenic causes of stroke are rare but should not be missed by the neurologist. The purpose of this review is to aid the reader in the evaluation of a patient with cryptogenic stroke with or without a family history suspicious for an inherited condition. The clinical findings, diagnosis, and management of monogenic causes of stroke and stroke look-alikes are discussed, including cerebral autosomal-dominant arteriopathy with subcortical infarcts and leukoencephalopathy, Fabry's disease, vascular Ehlers-Danlos, Marfan syndrome, sickle cell disease, the thrombophilias, hereditary hemorrhagic telangiectasia, cerebral cavernous malformations, hereditary cerebral hemorrhage with amyloidosis, and mitochondrial encephalopathy, lactic acidosis, and strokelike episodes. A quick review of systems designed to screen for genetic stroke causes is presented. By correlating stroke subtype with phenotype, this review will familiarize the clinician with indications for focused genetic testing in appropriate patients. PMID- 16479443 TI - Ischemic stroke as a complex genetic disorder. AB - Ischemic stroke is a complex genetic disorder, diverse in pathophysiology and clinical presentation. This phenotypic diversity may reflect genotypic diversity. One approach to discovering genetic risk factors has been to study intermediate phenotypes as in carotid intima-media thickness and leukoaraiosis. Intermediate phenotypes can be highly heritable, assessed in stroke-affected and unaffected individuals, and analyzed as quantitative traits. Because there is not a one-to one relationship between intermediate phenotypes and ischemic stroke, polymorphisms or haplotypes associated with an intermediate phenotype should be validated as risk factors for stroke itself. A complementary approach to gene discovery has been to fractionate the ischemic stroke phenotype into more homogeneous subphenotypes. Family history studies suggest that some subphenotypes like cardioembolic stroke may be less heritable than others. PMID- 16479444 TI - Disorders of coagulation in stroke. AB - The roles of the coagulopathies in thrombosis have been well established; however, the role in ischemic stroke is poorly understood. This article reviews the recent advances in the genetics of coagulation, the recently identified interactive roles of the coagulation and anticoagulation proteins, and the literature supporting protein dysregulation as a causative agent in cerebral venous thrombosis and ischemic stroke. PMID- 16479445 TI - Vascular biology: cellular and molecular profiling. AB - Our understanding of the mechanisms underlying cerebrovascular atherosclerosis has improved in recent years, but significant gaps remain. New insights into the vascular biological processes that result in ischemic stroke may come from cellular and molecular profiling studies of the peripheral blood. In recent cellular profiling studies, increased levels of a proinflammatory T-cell subset (CD4 (+)CD28 (-)) have been associated with stroke recurrence and death. Expansion of this T-cell subset may occur after ischemic stroke and be a pathogenic mechanism leading to recurrent stroke and death. Increases in certain phenotypes of endothelial cell microparticles have been found in stroke patients relative to controls, possibly indicating a state of increased vascular risk. Molecular profiling approaches include gene expression profiling and proteomic methods that permit large-scale analyses of the transcriptome and the proteome, respectively. Ultimately panels of genes and proteins may be identified that are predictive of stroke risk. Cellular and molecular profiling studies of the peripheral blood and of atherosclerotic plaques may also pave the way for the development of therapeutic agents for primary and secondary stroke prevention. PMID- 16479446 TI - How and when do we alter inflammatory mechanisms in stroke? Will it help? AB - The emerging recognition of atherosclerosis as primarily a chronic inflammatory process opens new possibilities to the treatment options that can reduce the risk of atherothrombotic stroke. The question is, "How do we alter the inflammatory mechanisms in atherosclerosis and will it reduce the risk of stroke?" Investigation into the fundamental pathophysiological mechanisms involved in the initiation, formation, and activation of atherosclerotic plaques is critical if we are to successfully employ pharmacotherapeutic strategies to reduce stroke risk. This article will review the key inflammatory elements associated with atherosclerosis and atherothrombotic stroke, identify potential inflammatory markers associated with the increased risk of stroke, review the effects of currently available medications that may have a beneficial impact on atherosclerosis and stroke, and briefly explore potential future strategies for reducing inflammation. PMID- 16479447 TI - Do common infections cause stroke? AB - Infections have long been recognized as a potential, if uncommon, cause of cerebrovascular disease. In recent years, with the growing recognition of the importance of inflammation in atherosclerosis, there has been renewed interest in the possibility that common infections may participate in the atherosclerotic process or lead to stroke through other mechanisms. Specific organisms that have been implicated include Chlamydia pneumoniae, herpes viruses, human immunodeficiency virus, Helicobacter pylori, and organisms associated with periodontal infections. This article outlines the epidemiological, pathological, and laboratory evidence that these infections may be associated with atherosclerosis and stroke. Although definitive proof of an association between a specific infection and stroke is generally lacking, the accumulating evidence does indicate that several types of infections may be among the modifiable risk factors that contribute to the risk of stroke. PMID- 16479448 TI - Diabetes: impact on stroke risk and poststroke recovery. AB - The prevalence of diabetes has been increasing substantially each year. Thus, diabetes has become one of the most important risk factors for incident and recurrent ischemic strokes. The risk conferred by diabetes is higher for diabetic patients who are less than age 65 and is proportionally higher in race-ethnic minorities where diabetes prevalence is higher than for Caucasians. To reduce risk of ischemic stroke, a structured program that addresses all of the diabetes associated risk factors of the metabolic syndrome is likely to be most effective. Diabetes increases poststroke mortality and adversely affects poststroke outcomes as compared with stroke patients without diabetes. Further research is needed to clarify optimal ways of reducing mortality and improving outcomes. PMID- 16479449 TI - Current understanding of multiple risk factors as the metabolic syndrome: distillation or deconstruction. AB - The "metabolic syndrome" is a new term that defines the clustering of vascular risk factors, such as hyperlipidemia, obesity, elevated blood pressure, and elevated blood glucose. Controversy exists regarding the use of the term, which raises the question of whether the unique grouping of vascular risk factors adds more clinical risk then the additive effect of multiple risk factors viewed as separate but important entities. Whatever the answer, the metabolic syndrome constitutes a major public health problem with over 47 million persons in the United States meeting criteria for the metabolic syndrome. Although studies have demonstrated that the metabolic syndrome is a risk factor for overall mortality as well as cardiovascular events, the relationship between the metabolic syndrome and ischemic stroke has not been well characterized. Two large cross-sectional studies report an association between metabolic syndrome and increased risk of a history of stroke. One large multiethnic prospective study found the metabolic syndrome to be significantly associated with an increased risk of ischemic stroke after adjustment for sociodemographics and other cardiovascular risk factors. This study estimated that the metabolic syndrome may account for 19% of ischemic strokes including 30% of stroke in women and over 40% of stroke in Hispanics. Despite debate about the utility of its definition, there is evidence to suggest that the metabolic syndrome is an important risk factor for ischemic stroke, with differential effects by gender and race-ethnicity. Further, the metabolic syndrome has important clinical and public health implications by helping to easily identify individuals at greatest risk of vascular events. PMID- 16479450 TI - Sleep disorders and stroke. AB - Obstructive sleep apnea (OSA) is a very common condition in patients with stroke and is found in over half of stroke patients. There is a complex relationship between OSA and stroke, attributable to shared risk factors. There are numerous mechanisms by which OSA may contribute to increased stroke risk, including promotion of atherosclerosis, hypercoagulability, and adverse effects on cerebral hemodynamics. Obstructive sleep apnea is also a risk factor for hypertension, and likely for atrial fibrillation and diabetes, conditions that in turn are risk factors for stroke. OSA is also associated with poor outcomes following stroke. Further epidemiological studies are needed to assess the relationship between OSA and stroke better. Clinical trials using continuous positive airway pressure as a treatment for OSA in stroke patients are needed to determine whether treatment of this condition alters outcome following stroke. PMID- 16479451 TI - Hormone replacement therapy and stroke: the current state of knowledge and directions for future research. AB - The Women's Health Initiative, which reported an increased risk of stroke with hormone therapy (HT), has had a major impact on the use of these drugs. The increased risk was unexpected because experimental animal studies showed estrogen reduced stroke volume and estrogen improves endothelial-dependent blood flow and improves lipid profiles in humans. The mechanisms of risk are uncertain because of the complexity of actions of estrogen in different doses, with or without progesterone, and the presence or absence of atherosclerotic risk factors. Therefore, future research should focus on identifying the mechanism(s) of risk, determining the differential impact of estrogen on stroke compared with heart disease, and identifying women with vasomotor symptoms who may be at risk of stroke with short-term HT use. PMID- 16479452 TI - Practice-based clinical research and ethical decision making--Part I: deciding whether to incorporate practice-based research into your clinical practice. AB - Calls for increasing clinical research in routine care settings aim to improve the empirical and ethical foundations of evidence-based practice. In the treatment and prevention of stroke many troubling management questions remain uninvestigated by the clinical research enterprise. Many practicing physicians perceive available data as having limited applicability to their average patients. Recent efforts by the National Institute of Neurological Disorders and Stroke seek to establish clinical research networks that will expand the role of private practice physicians in the conduct of research on stroke and other neurological diseases, akin to practice-based research networks supported by the Agency for Healthcare Research and Quality and other National Institutes of Health institutes. Adding clinical research to one's practice of medicine has practical and ethical implications. Furthermore, because of the growth and expansion of clinical research, all practicing physicians should have a fundamental understanding of clinical research and the differences between clinical care and clinical research, even if they are not directly involved in the conduct of clinical research, to appropriately advise their patients. In this and the following article, we offer two practical frameworks to aid physicians contemplating adding clinical research to their everyday practice of medicine. PMID- 16479453 TI - Practice-based clinical research and ethical decision making--Part II: deciding whether to host a particular research study in your practice. AB - Due to the growth and expansion of clinical research, particularly research conducted in practice-based settings, all practicing physicians should have a fundamental understanding of clinical research and the differences between clinical care and clinical research. Physicians considering adding clinical research to their everyday practice of medicine should carefully assess the practical and ethical dimensions of this decision on their practice. We discuss a framework that can help physicians assess a study they are considering hosting in their practice for seven requirements for ethical clinical research. Such an assessment can help them determine whether they are comfortable that the study is ethical and whether they can adhere to the requirements of the protocol in an ethically appropriate manner to maintain the ethics of the study. Together with the systems-informed professionalism framework presented in the preceding article, using the seven requirements of ethical clinical research framework can help practicing physicians with ethical decision making regarding incorporating practice-based research into their everyday practice of medicine. PMID- 16479454 TI - Subarachnoid hemorrhage. PMID- 16479456 TI - Tissue factor. PMID- 16479457 TI - Tissue factor in hemostasis and thrombosis. AB - Tissue factor (TF) plays an essential role in hemostasis. The tissue-specific pattern of TF expression is consistent with additional hemostatic protection in vital organs. An aberrant expression of TF within the vasculature occurs in a variety of diseases, including atherosclerosis, cancer, and sepsis. TF expression in these diseases is associated with thrombotic events. Future therapeutic strategies may prove beneficial in the treatment of thrombosis. However, these strategies should be designed to avoid compromising hemostasis. PMID- 16479458 TI - Sources of tissue factor. AB - Tissue factor (TF) exhibits a distinct nonuniform tissue distribution. Thus, high levels are found in highly vascularized organs such as the lung, brain, and placenta; intermediate levels in the heart, kidney, intestine, testes, and uterus; and low levels in the spleen, thymus, and liver. Several cell types are known to express TF constitutively, such as astrocytes in the brain, epithelial cells enveloping organs and body surfaces, adventitial fibroblasts and pericytes, and cardial myocytes in the heart. Smooth muscle cells in the media of the vessel wall and monocytes/macrophages contain small amounts of TF, which is enhanced substantially upon activation of the cells. Endothelial cells probably do not express TF. The popular concept of TF serving predominantly as a hemostatic envelope encapsulating the vascular bed has been challenged recently by the observation that blood of healthy individuals may form TF-induced thrombi under conditions entailing shear stress and activated platelets, corroborating the notion of blood-borne TF. Accordingly, small amounts of decrypted TF activity is detected in calcium ionophore-stimulated monocytes, and microparticles from plasma of healthy subjects possess TF-like activity subject to partial inactivation by anti-TF antibody. In addition to microparticles, plasma TF also comprises the soluble alternatively spliced human TF and truncated TF, both of which probably require factor VIIa to be physiologically active. Although it has been suggested that activated platelets possess active TF, the notion of TF as an integral platelet component is contested by more recent data. Rather, platelets may be very important in decrypting monocyte TF activity in a process entailing transfer of TF to activated platelets. PMID- 16479460 TI - Tissue factor in infection and severe inflammation. AB - In the pathogenesis of vascular disease, inflammation and coagulation play a pivotal role. Increasing evidence points to an extensive cross-talk between these two systems, whereby inflammation not only leads to activation of coagulation, but coagulation also considerably affects inflammatory activity. Tissue factor (TF) plays an important role at the crossroad of coagulation and inflammation, as the principal initiator of coagulation and an important modulator of inflammation. Proinflammatory cytokines can induce TF expression on mononuclear cells and endothelial cells and thereby commence pathways that lead to thrombin generation. Simultaneously, TF may bind to cellular receptors, which may affect the production and release of inflammatory mediators. There is increasing experimental evidence that TF inhibition may have beneficial effects in disease states in which the combination of coagulation and inflammation plays a prominent role. PMID- 16479459 TI - Emerging insights in tissue factor-dependent signaling events. AB - The complex of the cell surface receptor tissue factor (TF) and its ligand coagulation factor VIIa (FVIIa) is the primary initiator of the coagulation cascade. It is now clear the TF-initiated coagulation pathway also plays major nonhemostatic roles in inflammation, tumor growth, and angiogenesis. Direct or indirect cell signaling by TF-FVIIa or downstream coagulation proteases is an essential part of these nonhemostatic functions. The TF-FVIIa complex activates protease-activated receptor 2 and thus regulates gene transcription and protein translation, cell proliferation and survival, or cell motility and integrin activation. In this review, we relate our current understanding of direct TF signaling pathways to the emerging roles of TF in (patho)physiology. PMID- 16479461 TI - Tissue factor expression in the morphologic spectrum of vulnerable atherosclerotic plaques. AB - Inflammation and thrombosis are key events in the long-lasting sequence of atherosclerotic plaque initiation, plaque growth, and eventual onset of complications leading to clinically manifest disease. Recent cellular and molecular studies have indicated that inside the plaque tissue complex, proinflammatory and prothrombotic mechanisms are intimately associated, and tissue factor (TF) is one of the main proteins that may link both processes. It is therefore not surprising that TF expression appeared to be a prominent feature in various types of vulnerable atherosclerotic plaques (i.e., lesions that specifically predispose to the onset of symptomatic atherosclerotic disease). PMID- 16479462 TI - Tissue factor in trauma and organ dysfunction. AB - Tissue factor (TF) performs an essential role in the blood clotting system by activating the extrinsic coagulation pathway following vascular injury. In addition to this physiological hemostatic role for wound repair, TF also plays pivotal roles in organ dysfunction in trauma patients by triggering pathological disseminated thrombosis and inflammation. Constitutively expressed TF in subendothelial cells is released into the circulation following trauma and can be detected as slightly elevated TF levels in the plasma. Liberation of constitutive TF into the blood and inducible tissue factor expression on monocytes and the other cells may synergistically increase plasma TF levels to higher values at the early stage of posttrauma, especially in patients with disseminated intravascular coagulation (DIC) in association with sustained systemic inflammatory response syndrome. Marked TF generation not adequately balanced by physiological coagulation inhibitors such as tissue factor pathway inhibitor in posttrauma DIC patients has been observed. Based on these pieces of evidence, it has now been accepted that combined activation of TF-dependent coagulation inadequately regulated by anticoagulant mechanisms and inflammation may synergistically play important roles in the pathogenesis of posttrauma multiple organ dysfunction syndrome. PMID- 16479463 TI - Tissue factor in cancer and angiogenesis: the molecular link between genetic tumor progression, tumor neovascularization, and cancer coagulopathy. AB - Tissue factor (TF), the primary cellular initiator of blood coagulation, is also involved in cancer-related processes such as hypercoagulability (Trousseau syndrome), tumor growth, angiogenesis, and metastasis. Indeed, elevated TF expression by cancer cells and their associated endothelial cells has been reported frequently. Oncogenic events in cancer cells (e.g., expression of mutant K- ras, EGFR, PTEN or p53) lead to an increase in TF levels and activity, and thereby promote tumor aggressiveness, angiogenesis, and hypercoagulability. Like TF, thrombin receptor (protease-activated receptor-1) is also upregulated in cancer cells expressing oncogenic K -ras. Pharmacological antagonists of some of these transforming genes (e.g., epidermal growth factor receptor inhibitors) could diminish TF expression, both locally and systemically, and hence these targeted agents could be viewed as potential indirect and cancer-specific anticoagulants, in addition to their direct antitumor effects. We postulate that levels of circulating TF may be useful in monitoring the biological activity of these agents. Although TF is essential for vascular development, its expression by tumor-associated endothelium appears to play a subtle and seemingly dispensable role. Thus, TF is a pivotal element of the tumor-vascular interface, is involved in many cancer-related processes, and may well constitute a promising new target for anticancer combination therapies in some disease settings. PMID- 16479464 TI - Pharmacological inhibition of tissue factor. AB - Tissue factor plays an essential role in the initiation of coagulation in vivo. In severe conditions, including sepsis and acute lung injury, increased expression of tissue factor may induce disseminated intravascular coagulation and fibrin deposition in organs, which are believed to have a determining impact on patient outcome. Tissue factor also acts as a signaling receptor and is involved in the systemic inflammatory response, as in cancer progression and atherosclerosis. Interventions aiming at limiting tissue factor activities have been evaluated in multiple experimental studies and the observed results have supported the potential benefits for coagulation disorders, inflammation, and survival. The effects of the main physiological inhibitor of tissue factor, tissue factor pathway inhibitor, have been evaluated in two large clinical trials in sepsis. Even though they are not associated with an improved outcome, the observed data support further clinical studies. PMID- 16479466 TI - [Evaluation of the medical curriculum: who should be asked and when about the clinical relevance?]. PMID- 16479467 TI - [Potential improvements in medical education as retrospectively evaluated by candidates for specialist examinations]. AB - BACKGROUND AND OBJECTIVE: As part of the new regulations for licensing doctors there have been numerous attempts at reform by many medical faculties to consider interdisciplinary linkage of the curriculum with emphasis on teaching of small groups of students. This study was undertaken to help answer the question of how much weight should be given to the various subjects and what resources are needed for any reformed curriculum and what key areas of competence need to be given greater importance. METHODS: 1029 candidates of specialist examinations of the Medical Council of North-Rhine in 2002 and 2003 filled in questionnaires to evaluate retrospectively the actual relevance of individual preclinical and clinical subjects, courses and areas of practical competence to their further medical education and related potentials for improvement in their studies. The participants were from 5 medical faculties in the North-Rhine area of Germany. They were also asked about methods of examination that were effective in aiding their learning behaviour. RESULTS: Those answering the questionnaire considered especially chemistry and physics as well as environmental, occupational and forensic medicine, bio-mathematics, radiotherapy and nuclear medicine among the less relevant subjects. On the other hand, anatomy, physiology, internal medicine, pharmacology and surgery were considered especially relevant. CONCLUSION: The greatest deficiencies in most of the medical curricula as taught in the North-Rhine medical courses are in the areas of competence in communication and practical clinical skills. Members of this group also pleaded for an increased use of standardized objective structured clinical examinations (OSCE). PMID- 16479468 TI - [Results of coloscopy screening in 2005--an Internet-based documentation]. AB - BACKGROUND AND OBJECTIVE: In October 2002 screening coloscopy was introduced into the National Cancer Prevention Programme in Germany. The results of an online registry are presented here. METHODS: Data from consecutive screening colonoscopies in the practices of the 280 participating gastroenterologists, performed in asymptomatic subjects, were collected in an online registry. Number and histology of colorectal polyps and carcinomas, complication rates of colonoscopy and polypectomy were registered. Advanced adenoma was defined as an adenoma >10 mm in diameter, with villous or tubulovillous histology, or presence of high-grade dysplasia. RESULTS: A total of 109989 colonoscopies (43% in males) were evaluated from October 2003 to July 2005. Tubular and villous adenomas were found in 16.2% and 3.8%, respectively, whereas invasive cancers were diagnosed in 0.7%. Advanced adenomas amounted to 6.1%.The majority of carcinomas were detected in early stages (UICC stages I and II in 48 and 22 %, respectively). -In most of the polyps immediate polypectomy was carried out. The complication rate was low and no deaths were observed: cardiopulmonary complications occurred in 0.10% of the colonoscopies, bleeding in 0.79% of polypectomies most of which were managed endoscopically (surgery in 0.04% of polypectomies). Perforation occurred in 0.02% of the colonoscopies and 0.10% of polypectomies. CONCLUSIONS: Neoplasias of the colon were detected in about 20% of persons who had taken part in a colonoscopy screening programme: most of the lesions were immediately removed by polypectomy. The high rate of early stages of colorectal cancers detected by screening colonoscopy is an indirect indicator of mortality reduction. In Germany screening colonoscopy has a low risk. PMID- 16479469 TI - [Disseminated herpes zoster in diabetes mellitus]. AB - HISTORY AND ADMISSION FINDINGS: A 71-year old man presented with painful hemorrhagic vesicles and papules over the entire body that had persisted for three days. Type 2 diabetes mellitus type 2 had been diagnosed 20 years ago and had not been treated for the last 5 years. Therapy had been discontinued by the patient. INVESTIGATIONS: HbA1c (11,9%) and blood glucose levels (up to 360 mg/dl) were abnormal. Varicella-zoster-DNA was replicated by PCR from the vesicle fluid. DIAGNOSIS AND TREATMENT: After the clinical diagnosis of disseminated herpes zoster had been confirmed systemic therapy with aciclovir 10 mg/kg day was started. There was no evidence of malignancy. Insulin therapy was initiated. CONCLUSION: Dissemination is a rare complication of herpes zoster, aided by immunosuppression. In the presented case there was no evidence of malignancy or other cause of immunosuppression, but the patient also had type 2 diabetes with very high blood glucose levels. The diabetes was thought to be causally related to the ineffective immune response to varicella zoster virus. There has been no previous published report of this relationship. PMID- 16479471 TI - [Bullous pemphigoid: diagnostics and new therapeutic strategies]. AB - Bullous pemphigoid, the most frequent bullous autoimmune dermatosis of the adult, typically presents as disseminated tense blisters on normal or erythematous skin. The diagnosis can be confirmed by direct and indirect immunofluorescence, the detection of circulating autoantibodies against the basement membrane proteins collagen XVII/BP180 and BP230, and histopathology. Autoantibody reactivity against collagen XVII can be measured by ELISA and correlates with disease activity. The ELISA therefore provides a useful tool for monitoring disease activity. Treatment of bullous pemphigoid usually consists of topical and / or systemic steroids in combination with immunosuppressive agents. The intensity of skin involvement and the concurrent diseases and medications of the patient must be considered when selecting a certain treatment. Interdisciplinary cooperation between general practitioners, internists and other specialists facilitates the optimal adaptation of the medication and the early discovery of potential side effects. PMID- 16479472 TI - [Obesity-induced respiratory dysfunction in children and juveniles]. PMID- 16479473 TI - [Duty of the physician and the hospital to inform the patient]. PMID- 16479474 TI - [Recommendations for peri-procedural thrombocyte aggregation inhibition]. PMID- 16479477 TI - Morphomechanics: goals, basic experiments and models. AB - Morphomechanics is a branch of developmental biology, studying the generation, space-time patterns and morphogenetic role of mechanical stresses (MS) which reside in embryonic tissues. All the morphogenetically active embryonic tissues studied in this respect have been shown to bear substantial mechanical stresses of tension or pressure. MS are indispensable for organized cell movements, expression of a number of developmentally important genes and the very viability of cells. Even a temporary relaxation of MS leads to an increase in the morphological variability and asymmetry of embryonic rudiments. Moreover, MS may be among the decisive links of morphogenetic feedback required for driving forth embryonic development and providing its regular space-time patterns. We hypothesize that one such feedback is based upon the tendency of cells and tissues to hyperrestore (restore with an overshoot) their MS values after any deviations, either artificial or produced by neighboring morphogenetically active tissues. This idea is supported by a number of observations and experiments performed on the tissue and individual cell levels. We describe also the models demonstrating that a number of biologically realistic stationary shapes and propagating waves can be generated by varying the parameters of the hyperrestoration feedback loop. Morphomechanics is an important and rapidly developing branch of developmental and cell biology, being complementary to other approaches. PMID- 16479478 TI - Direct physical formation of anatomical structures by cell traction forces. An interview with Albert Harris by Lev Beloussov. AB - Albert Harris was educated at The Norfolk Academy, Norfolk, Virginia, USA (1961). He then earned a Batchelor of Arts Degree in Biology from Swarthmore College, in Pennsylvania, USA (1965), followed by a Ph.D. in Biology (1971) from Yale University, where his Dissertation Advisor was the great John Phillip Trinkaus. He held a Damon-Runyon Postdoctoral Fellowship in Cancer Research in 1970-72, under Michael Abercrombie, FRS, at the Strangeways Research Laboratory of Cambridge University, England. Then he accepted a position as Assistant Professor in the Zoology Department of the University of North Carolina at Chapel Hill, N.C. USA. In 1977, he was promoted to Associate Professor of Zoology, and in 1983 was promoted to Full Professor of Biology. In Oct.-Nov. 1991 he was honored to be Distinguished Visiting Professor of Zoology at the University of California at Davis. PMID- 16479479 TI - From observations to paradigms; the importance of theories and models. An interview with Hans Meinhardt by Richard Gordon and Lev Beloussov. AB - Hans Meinhardt received his PhD in physics from the University of Cologne at 1966. For a postdoctoral fellowship, he went to the European High Energy Laboratory CERN in Geneva where he joined a group working on the leptonic decay of the Xi-minus particle. One of his duties was to perform computer simulations to optimize the complex experimental setup--a skill which turned out to be helpful later on. In 1969 he switched to biology and joined the department of Alfred Gierer at the Max Planck Institute for Developmental Biology (formerly Virus Research) in Tubingen. His interest was focused on mechanisms of biological pattern formation. Using computer simulations as a tool, he developed models for essential steps in development. Most fascinating for him was the possibility to recapitulate and to reconstructusing the computer the genesis of structures where no structures were before and to see how these emerging structures become subsequently further refined. In addition to the interaction with Alfred Gierer and his group working on hydra development, the Max-Planck Institute as a whole provided a very stimulating environment. In the seventies, the work of Klaus Sander on gradients in early insect development was highly influential. Collaboration with Martin Klinger in the eighties revealed that the pigmentation patterns on tropical sea shells are convenient to study highly dynamic patterning processes. The variability and the asthetic beauty of these patterns turned out to result from the chaotic nature of the underlying reactions. Mechanisms deduced from shell patterns became a key to understand other developing systems such as orientation of chemotactic cells or phyllotaxis. Officially Hans Meinhardt retired at the end of 2003. At present he works on refinements and extensions of models which account for the different modes of embryonic axis formation in different phyla from an evolutionary point of view. PMID- 16479480 TI - Gastrulation in amphibian embryos, regarded as a succession of biomechanical feedback events. AB - Gastrulation in amphibian embryos is a composition of several differently located morphogenetic movements which are perfectly coordinated with each other both in space and time. We hypothesize that this coordination is mediated by biomechanical interactions between different parts of a gastrulating embryo based upon the tendency of each part to hyper-restore the value of its mechanical stress. The entire process of gastrulation in amphibian embryos is considered as a chain of these mutually coupled reactions, which are largely dependent upon the geometry of a given embryo part. We divide gastrulation into several partly overlapped steps, give a theoretical interpretation for each of them, formulate the experiments for testing our interpretation and describe the experimental results which confirm our point of view. Among the predicted experimental results are: inhibition of radial cell intercalation by relaxation of tensile stresses at the blastula stage; inversion of convergent intercalation movements by relaxation of circumferential stresses at the early gastrula stage; stress-promoted reorientation of axial rudiments, and others. We also show that gastrulation is going on under a more or less constant average value of tensile stresses which may play a role as rate-limiting factors. A macro-morphological biomechanical approach developed in this paper is regarded as complementary to exploring the molecular machinery of gastrulation. PMID- 16479481 TI - Principles of branch formation and branch patterning in Hydrozoa. AB - The freshwater polyp Hydra produces buds which separate from the parent. Other Hydrozoa produce branches which remain connected to the parent, thus forming a colony. Some Hydrozoa grow by means of an organ that is like a shoot apical meristem. Others display a sympodial type of growth. In this article, I propose that these different types of branches are organized by a common pattern-forming system. This system has self-organizing properties. It causes branch tip formation and is kept active in the tip when the tip finally differentiates into a hypostome of a polyp. The system does not cause structure formation directly but rather, determines a tissue property called positional value, in such a way that a gradient of values forms in the tissue of the bud or branch. The local value determines the local morphodynamic processes, including differentiation of the hypostome (highest positional value), tentacles and basal disc and of the exoskeleton pattern along the shoot. A high positional value favors the onset of a new self-organizing process and by lateral inhibition, such a process prevents the initiation of a further process in its surroundings. Small quantitative differences in the range of the signals involved determine whether a bud or a branch forms and whether monopodial and sympodial growth follows. PMID- 16479482 TI - A hypothesis linking low folate intake to neural tube defects due to failure of post-translation methylations of the cytoskeleton. AB - Neural tube defects are serious congenital malformations which can be prevented by periconceptional folic acid supplementation. We hypothesize that folic acid provides the methyl group used for post-translational methylation of arginine and histidine in the highly conserved regulatory domains of the cytoskeleton and that these are required for neural tissue differentiation. Presumptive neural tissue has an unusually high need for folates due to the activity of phosphoethanolamine methyl transferase in producing neural tissue specific lipids at a time when the cytoskeleton is also competing for methylation. According to the cell state splitter hypothesis, the cytoskeleton is required to coordinate the spatial and temporal component of differentiation. When folate supply is low and the cytoskeleton is not methylated properly, the result is a neural tube defect due to failure of this coordination. PMID- 16479483 TI - Blastula wall invagination examined on the basis of shape behavior of vesicular objects with laminar envelopes. AB - A complete description of the blastula shape transformations which occur in the process of gastrulation involves the mechanics of the blastula wall. It is inferred that the mechanical properties of this wall can, to a first approximation, be described in terms of elastic deformational modes which are equivalent to those of simple vesicular systems such as phospholipid vesicles and red blood cells. For membranes composed of an arbitrary number of aligned, laterally unconnected thin layers, these are area expansivity and local and non local bending. Stable shapes of vesicular objects with laminar envelopes are defined as shapes which correspond to the minimum of the system elastic energy. The shape behavior of these objects is briefly reported. The criterion for the stability of the spherical shape of a vesicular object is derived. In particular we analyze the shape transition from a sphere to an invaginated gastrula. Conditions are demonstrated under which this shape transition can occur through a series of continuous shape transformations. A possible example of such a shape transformation is given. The blastula wall of the sea urchin is modeled as a closed laminar membranous system with three layers, the monocellular epithelial sheet and the extracellular apical lamina and hyaline layers. Experimentally measured elastic constants of some of these layers are shown to be in accord with the restrictions on their values imposed by the requirement for continuous shape transformations. PMID- 16479484 TI - Do lamellipodia have the mechanical capacity to drive convergent extension? AB - Convergent extension (CE), a kinematic motif associated with several important morphogenetic movements in embryos, entails narrowing of a tissue in one in-plane direction and elongation in the other. Although the cell elongation and intercalation which accompany this process have been investigated and relevant genes and biochemical pathways have been studied in multiple organisms, a fundamental question that has not yet been answered is "Do the lamellipodia thought to drive these motions actually have the mechanical capacity to do so?" Here, we address this and a number of related issues using a state-of-the-art computational model which can replicate cell motions, changes in cell shape and tissue deformations. The model is based on the cell-level finite element approach of Chen and Brodland, but has additional features which allow it to model lamellipodium formation and contraction. In studying CE, computational models provide an important complement to molecular approaches because they reveal the "mechanical pathways" through which gene products must ultimately act in order to produce physical movements. The model shows that lamellipodia can drive CE, that they do so through cell intercalations and that the elongated cells characteristic of CE arise only when adjacent tissues resist convergence, a result which we confirm experimentally. PMID- 16479485 TI - Geometry and mechanics of teleost gastrulation and the formation of primary embryonic axes. AB - Examination of normal shaping dynamics and immediate and long-term responses to blastoderm cutting in zebrafish and loach embryos prior to the onset of gastrulation and during the course of epiboly revealed that anteroposterior (AP) and dorsoventral (DV) polarity formation is connected with shaping of the blastoderm circumferential region, which stretches along and shrinks across its movement axes and originates the non-isotropic fields of tensile stresses. Based on data from cutting experiments and quantitative morphology, we reconstructed the movement-shaping patterns of epiboly and embryonic shield formation. We revealed that AP and DV axes originate as a mass cell movement subject to the movement-shaping equivalence principle, which means the spatial series of differently shaped areas corresponding to the time succession of the same area shaping. Maintenance of the main body axes in orthogonal orientation depends on the mechanical equilibrium principle allowing for converting shape asymmetry into that of tensile stresses and vice versa. The causal relationship between the main movement-shaping axes and that of embryonic polarity was proved in cutting experiments in which the DV axis direction was subject to rearrangement so as to adjust to the new direction of mass cell movement axes induced by healing the wound in the blastoderm circumferential region. PMID- 16479486 TI - The dynamic geometry of mass cell movements in animal morphogenesis. AB - There is an infinite number of interactions between morphogenetic processes of different time and space scales. How do these unfold in a regular series of mass morphogenetic movements to produce a basically simple and reproducible structure? I present a new morphogenetic concept -- the spatial unfolding (SU) of cell movements, whose definition rests on the correspondence between the continuous spatial series of cell shapes and the succession of changes in the shape of a single cell moving in an epithelial sheet whose shape is also subject to change. The change in the shape of moving cells is the only measure of their translocation both in space and time. The SU provides a morpho-dynamics description of mass cell movements which is completely independent of both an external coordinate system and external forces. The cell geometry of SU allows us to derive the future embryonic form from the actual one by a movement-shaping algorithm operating on the basis of positive and negative geometric feedbacks between the cell movement in the epithelial sheet plane and the epithelial sheet shaping, the feedback system providing a geometric alternative to Turing's self organization via reaction-diffusion systems. Putting together histological, quantitative morphological and experimental data permits us to isolate four SU, each acting in morphogenesis as an irreducible whole, which seem to include all real examples of epithelial morphogenesis in multicellular animals, from Coelenterates to Chordates. PMID- 16479487 TI - Effects of microgravity on cell cytoskeleton and embryogenesis. AB - The aim of this review is to compile, summarize and discuss the effects of microgravity on embryos, cell structure and function that have been demonstrated from data obtained during experiments performed in space or in altered gravity induced by clinostats. In cells and tissues cellular structure and genetic expression may be changed in microgravity and this has a variety of effects on embryogenesis which include death of the embryo, failure of neural tube closure, or final deformities to the overall morphology of the newborn or hatchling. Many species and protocols have been used for microgravity space experiments making it difficult to compare results. Experiments on the ways in which embryonic development and cell interactions occur in microgravity could also be performed. Experiments that have been done with cells in microgravity show changes in morphology, cytoskeleton and function. Changes in cytoskeleton have been noted and studies on microtubules in gravity have shown that they are gravity sensitive. Further study of basic chemical reactions that occur in cells should be done to shed some light on the underling processes leading to the changes that are observed in cells and embryos in microgravity. PMID- 16479488 TI - On the origin of pattern and form in early Metazoans. AB - The 'Cambrian explosion', about 540 million years ago, may have occurred within 10 to 50 million years. Almost all of the modern phyla, a very restricted group and many groups that may represent extinct phyla, suddenly appear near that time in the fossil record. Numerous extensive periods of mass extinction since that time led to no new phyla. This is taken as an impetus to examine a possible source, beyond Darwinian adaptation, of the apparently restricted number of phyla. Such a postulated constraint or restriction beyond adaptation is proposed to be based on a mutation or mutations allowing single celled or colonial precursors to begin forming into epithelial sheets and gene activation patterns of a particular kind, those giving rise to the very earliest metazoans. The interaction of signaling pathways in pairs, with different pairs acting sequentially are proposed as key to this earliest patterning, such patterning being extensively elaborated over the last approximately 550 million years. Restrictions on the very large set of possible forms and patterns on which adaptation acts are discussed. PMID- 16479489 TI - An anisotropic-viscoplastic model of plant cell morphogenesis by tip growth. AB - Plant cell morphogenesis depends critically on two processes: the deposition of new wall material at the cell surface and the mechanical deformation of this material by the stresses resulting from the cell's turgor pressure. We developed a model of plant cell morphogenesis that is a first attempt at integrating these two processes. The model is based on the theories of thin shells and anisotropic viscoplasticity. It includes three sets of equations that give the connection between wall stresses, wall strains and cell geometry. We present an algorithm to solve these equations numerically. Application of this simulation approach to the morphogenesis of tip-growing cells illustrates how the viscoplastic properties of the cell wall affect the shape of the cell at steady state. The same simulation approach was also used to reproduce morphogenetic transients such as the initiation of tip growth and other non-steady changes in cell shape. Finally, we show that the mechanical anisotropy built into the model is required to account for observed patterns of wall expansion in plant cells. PMID- 16479490 TI - Morphogenesis, plasticity and irreversibility. AB - The dynamical systems theory of morphogenesis is surveyed, in which positional information is generated through intracellular reaction dynamics and cell-cell interaction. Cells differentiate because of these dynamics. In addition, these differentiated cells form an ordered spatial pattern, which further stabilizes the cellular states, leading to robust morphogenesis and irreversibility in the differentiation. Induction, community effect, gastrulation and activin-controlled artificial tissue-genesis are discussed from the perspective of this theory and the relevance of dynamics of cellular plasticity is stressed. PMID- 16479492 TI - Mechanics in embryogenesis and embryonics: prime mover or epiphenomenon? AB - Mechanics is shown to be an important, perhaps central component to the differentiation and development of embryos. Mechanics of the nucleus may also be involved in determining which genes are expressed in a given cell. There are two major approaches at present to the mechanics of differentiation in embryos: morphomechanics and differentiation waves. These are compared in detail, to provide a starting point for future experimental work to bring them into one conceptual framework. This may rationalize the present cookbookery of stem cell production by placing it in the context of differentiation waves and the differentiation code. Embryonics, the realization of concepts from embryology in computer hardware and software, might be considerably enhanced by incorporating mechanical concepts of embryogenesis. Segmented robots, modular robotics, cellular microrobotics, flexible electronics, wearable computers, diatom nanotechnology and waves in active media point to a synthesis that we could call embryonic robotics. PMID- 16479491 TI - Biophysical regulation during cardiac development and application to tissue engineering. AB - Tissue engineering combines the principles of biology, engineering and medicine to create biological substitutes of native tissues, with an overall objective to restore normal tissue function. It is thought that the factors regulating tissue development in vivo (genetic, molecular and physical) can also direct cell fate and tissue assembly in vitro. In light of this paradigm, tissue engineering can be viewed as an effort of "imitating nature". We first discuss biophysical regulation during cardiac development and the factors of interest for application in tissue engineering of the myocardium. Then we focus on the biomimetic approach to cardiac tissue engineering which involves the use of culture systems designed to recapitulate some aspects of the actual in vivo environment. To mimic cell signaling in native myocardium, subpopulations of neonatal rat heart cells were cultured at a physiologically high cell density in three-dimensional polymer scaffolds. To mimic the capillary network, highly porous elastomer scaffolds with arrays of parallel channels were perfused with culture medium. To mimic oxygen supply by hemoglobin, culture medium was supplemented with an oxygen carrier. To enhance electromechanical coupling, tissue constructs were induced to contract by applying electrical signals mimicking those in native heart. Over only eight days of cultivation, the biomimetic approach resulted in tissue constructs which contained electromechanically coupled cells expressing cardiac differentiation markers and cardiac-like ultrastructure and contracting synchronously in response to electrical stimulation. Ongoing studies are aimed at extending this approach to tissue engineering of functional cardiac grafts based on human cells. PMID- 16479493 TI - Mechanical control of tissue morphogenesis during embryological development. AB - Twenty years ago, we proposed a model of developmental control based on tensegrity architecture, in which tissue pattern formation in the embryo is controlled through mechanical interactions between cells and extracellular matrix (ECM) which place the tissue in a state of isometric tension (prestress). The model proposed that local changes in the mechanical compliance of the ECM, for example, due to regional variations in basement membrane degradation beneath growing epithelium, may result in local stretching of the ECM and associated adherent cells, much like a "run-in-a-stocking". Cell growth and function would be controlled locally though physical distortion of the associated cells, or changes in cytoskeletal tension. Importantly, experimental studies have demonstrated that cultured cells can be switched between different fates, including growth, differentiation, apoptosis, directional motility and different stem cell lineages, by modulating cell shape. Experiments in whole embryonic organ rudiments also have confirmed the tight correlation between basement membrane thinning, cell tension generation and new bud and branch formation during tissue morphogenesis and that this process can be inhibited or accelerated by dissipating or enhancing cytoskeletal tension, respectively. Taken together, this work confirms that mechanical forces generated in the cytoskeleton of individual cells and exerted on ECM scaffolds, play a critical role in the sculpting of the embryo. PMID- 16479494 TI - Morphomechanical programming of morphogenesis in cnidarian embryos. AB - The factors governing the pattern formation process in the early morphogenesis of a marine colonial hydroid, Dynamena pumila, have been studied. Two different types of morphogenesis have been distinguished. Morphogenesis of the first type goes on via changes in cell shape and cell axis orientation, while morphogenesis of the second type is based upon the active coordinated cell movements associated with cell rearrangements. It was shown that morphogenesis of both types can be considered as cascades in which any event is a consequence of the previous one. The spatial structure of each developmental stage contains information about the direction and the initial conditions of further morphogenesis. So, an "epigenetic program" of morphogenesis gradually originates in the course of development and provides the stable reproduction of spatial structures. It is reasonable to consider the activity of epigenetic factors guiding Dynamena morphogenesis (geometry/topology of an embryo, heterogeneity of an embryo spatial structure, configuration of the field of mechanical stresses of the embryo surface) as "morphomechanical programming" of morphogenesis. PMID- 16479495 TI - Morphodynamics of phyllotaxis. AB - A morphodynamic model for phyllotaxis based upon an axiomatic approach is presented. We show that the helical forms of alternate phyllotaxis can be derived from the assumption of the rudiment's growth and movement on the cylindrical embryo surface in the absence of a longitudinal displacement. This leads to the repeating transition of tetragonal packaging of the rudiments into hexagonal packaging and vice versa. Under these conditions, sequences of rudiments produce either left-handed or right-handed helices, the number of which at the circumference of the cylinder corresponds to adjacent numbers of the Fibonacci series. Cross-opposite phyllotaxis forms are defined as superior with respect to the alternate ones, and verticillate phyllotaxis forms as superior with respect to the cross-opposite ones. Different phyllotaxis forms can be interpreted as a result of stretching of crystalline structures of the embryo formed by dense packing of rudiments. The superior phyllotaxis forms can be considered as the additive summation of lower forms. Morphodynamic mechanisms underlying the formation of multiple forms of helical phyllotaxis are discussed. PMID- 16479497 TI - Pulling forces acting on Hox gene clusters cause expression collinearity. AB - The development of normal patterns along the primary and secondary vertebrate axes depends on the regularity of early Hox gene expression. During initial stages, these expression events form a sequential pattern of partially overlapping domains along the anteroposterior axis in coincidence with the 3' to 5' order of the genes in the Hox cluster (spatial collinearity). In addition, the genes are activated one after the other in the 3' to 5'order (temporal collinearity). These features are poorly understood within the framework of Molecular Genetics. A model was proposed according to which physical forces act on Hox clusters as a result of signaling from morphogen gradients. The model can explain the collinearity of Hox gene expression along the primary and secondary body axes. The increase in the concentration of morphogen is accordingly followed by an increase of the force acting on the cluster. The genes are sequentially translocated, in the 3' to 5' order, toward the interchromosome domain where they are exposed to transcription factors for activation. The above geometrodynamic approach reproduces most collinearity data. Recent experiments verify the above prediction of sequential 3' to 5' Hox gene translocations in the interchromosome domain. Furthermore, it seems that these translocations, combined with cluster decondensations, are caused by attractive forces acting on the 3' end of the cluster and pulling the genes out of the chromosome territory. Additional experiments are proposed in order to specify the origin of the forces. PMID- 16479496 TI - Before programs: the physical origination of multicellular forms. AB - By examining the formative role of physical processes in modern-day developmental systems, we infer that although such determinants are subject to constraints and rarely act in a "pure" fashion, they are identical to processes generic to all viscoelastic, chemically excitable media, non-living as well as living. The processes considered are free diffusion, immiscible liquid behavior, oscillation and multistability of chemical state, reaction-diffusion coupling and mechanochemical responsivity. We suggest that such processes had freer reign at early stages in the history of multicellular life, when less evolution had occurred of genetic mechanisms for stabilization and entrenchment of functionally successful morphologies. From this we devise a hypothetical scenario for pattern formation and morphogenesis in the earliest metazoa. We show that the expected morphologies that would arise during this relatively unconstrained "physical" stage of evolution correspond to the hollow, multilayered and segmented morphotypes seen in the gastrulation stage embryos of modern-day metazoa as well as in Ediacaran fossil deposits of approximately 600 Ma. We suggest several ways in which organisms that were originally formed by predominantly physical mechanisms could have evolved genetic mechanisms to perpetuate their morphologies. PMID- 16479498 TI - Spatial patterns formed by chemotactic bacteria Escherichia coli. AB - In certain experimental conditions, bacteria form complex spatial-temporal patterns. A striking example of such kind was reported by Budrene and Berg (1991), who observed a wide variety of different colony structures ranging from arrays of spots to radially oriented stripes or arrangements of more complex elongated spots, formed by Escherichia coli. We discuss the relevant mechanisms of intercellular regulation in bacterial colony which may cause pattern formation, and formulate the corresponding mathematical model. In numerical experiments a variety of patterns, observed in real systems, is reproduced. The dynamics of their formation is investigated. PMID- 16479499 TI - The natural variability of morphogenesis: a tool for exploring the mechanics of gastrulation movements in amphibian embryos. AB - The investigation of natural variability of metric morphological characters in frog gastrulation revealed that in genetically and environmentally homogeneous samples of embryos their variability is of a higher order of magnitude than that known for quantitative metric characters in adult organisms. Matching the coefficients of variation of characters under consideration to the specific rates of their changes in normal development revealed a strong positive correlation between the rates of morphological change and the amount of morphological variance. The increase in the variance is mainly in characters concerned with shaping of moving embryonic areas and arises as a result of a positive feedback between the movement of a given area and recruitment of cells from surrounding areas into the movement. The account of natural variation suggests a new model of amphibian gastrulation whose essential feature is the intimate connection between the movement and shaping of the dorsal blastopore lip of the gastrula. PMID- 16479500 TI - Biophysical mechanisms of cardiac looping. AB - During early embryogenesis, the heart is a single, relatively straight tube which bends and twists (loops) rightward to create the basic plan of the mature heart. Despite intensive study for many decades, the biophysical mechanisms which drive and regulate cardiac looping have remained poorly understood. This review discusses, from a historical perspective, studies of looping mechanics and various theories which have been proposed for this complex process. Then, based on recently acquired data, a new biomechanical hypothesis is proposed for the rst phase of looping (c-looping). Understanding morphogenetic mechanisms would facilitate research devoted to preventing and treating congenital heart malformations caused by looping abnormalities. PMID- 16479501 TI - The role and limits of a gradient based explanation of morphogenesis: a theoretical consideration. AB - Development of an organism is a multi-dimensional process leading to the generation of complex species-specific structures. This specificity suggests machine-like organisation. The uneven distribution (gradient) of soluble substances (morphogens) and specific receptor-ligand interactions are known to cause differential gene expression. Therefore gradients of morphogens are used as a causal explanation of developmental processes. However each attempt to describe development causally should take into account both the local fine organisation and global robustness of morphogenesis. The classical view of the role of morphogens will be critically considered and possible alternative proposed. The core idea of my proposal is that the main function of morphogenetic substances could be a context dependent modification of cell behaviour. Both history and different features of morphogenetic fields create the framework for the activity of morphogenes. PMID- 16479502 TI - The evolution of the structure of tubulin and its potential consequences for the role and function of microtubules in cells and embryos. AB - This paper discusses the results of homology modeling and resulting calculation of key structural and physical properties for close to 300 tubulin sequences, including alpha, beta, gamma, delta and epsilon -tubulins. The basis for our calculations was the structure of the tubulin dimer published several years ago by Nogales et al. (1998), later refined to 3.5 resolution by Lowe et al. (2001). While, it appears that the alpha, beta and gamma-tubulins segregate into distinct structural families, we have found several differences in the physical properties within each group. Each of the alpha, beta and gamma- tubulin groups exhibit major differences in their net electric charge, dipole moments and dipole vector orientations. These properties could influence functional characteristics such as microtubule stability and assembly kinetics, due to their effects on the strength of protein-protein interactions. In addition to the general structural trends between tubulin isoforms, we have observed that the carboxy-termini of alpha and beta-tubulin exists in at least two stable configurations, either projecting away from the tubulin (or microtubule) surface, or collapsed onto the surface. In the latter case, the carboxy-termini form a lattice distinctly different from that of the well-known A and B lattices formed by the tubulin subunits. However, this C terminal lattice is indistinguishable from the lattice formed when the microtubule-associated protein tau binds to the microtubule surface. Finally, we have discussed how tubulin sequence diversity arose in evolution giving rise to its particular phylogeny and how it may be used in cell- and tissue-specific expression including embryonal development. PMID- 16479503 TI - Tissue morphogenesis: a surface buckling mechanism. AB - Surface patterns can emerge during growth of anisotropic tissues because of surface buckling. This morphogenetic scenario is examined in the present paper based on a simple phenomenological theory of tissue growth. In particular, we show that constrained growth can lead to tissue compression, which in turn may result in surface buckling of the tissue. The latter means the appearance of wavy patterns on the surface. These patterns decay away from the surface. It is interesting that the critical magnitude of the parameter of mass supply, which corresponds to surface buckling, is independent of the pattern wavelength and various patterns can generally be generated in growth. Results of theoretical analysis show that the surface buckling scenario is realistic if the growing tissue matches the following two conditions. First, compression should appear during tissue growth. Second, the tissue should exhibit strong anisotropy. The former condition does not necessarily mean geometric constraints: inhomogeneous growth or material inhomogeneity and anisotropy can lead to the appearance of compressive stresses. The latter condition is typical of some tissues with fiber reinforcement in planes parallel to the surface. In the latter case, the tissue material is much softer in the out-of-plane direction than in plane. The creation of patterns by restraining tissue growth and forcing the surface to buckle represents a challenging experimental problem. PMID- 16479505 TI - The clinical need for new antiviral drugs directed against influenza virus. PMID- 16479506 TI - Meningococcal control in the United States and Africa. PMID- 16479507 TI - The benefits and concerns related to vitamin a supplementation. PMID- 16479508 TI - Recovery of drug-resistant influenza virus from immunocompromised patients: a case series. AB - Influenza virus with resistance to antiviral drugs emerges with increased frequency in immunocompromised patients and can limit the benefit of M2 and neuraminidase (NA) inhibitors. We document 3 cases of influenza in severely immunocompromised patients from whom virus variants with molecular markers of resistance to anti-influenza drugs were recovered. Virus variants recovered from 2 patients had mutations in the M2, NA (with a previously recognized Glu119Val NA substitution), and hemagglutinin genes. We describe a novel Asp198Asn NA mutation in an influenza B virus and its decreased susceptibility to both oseltamivir and zanamivir. PMID- 16479509 TI - Comparative activities of oseltamivir and A-322278 in immunocompetent and immunocompromised murine models of influenza virus infection. AB - We developed an immunocompromised murine model of influenza virus infection and demonstrated comparable efficacy of oral oseltamivir and A-322278 (both given at dosages of 10 mg/kg/day) in reducing viral replication, decreasing weight loss, and prolonging survival. Once the treatment was discontinued, severe combined immunodeficient (SCID) mice had progressive viral replication and clinical decline. Drug-resistant variants were detected in 4 (29%) of 14 and 2 (13%) of 15 mice (both BALB/c and SCID) treated with oseltamivir or A-322278, respectively; no resistant variants were detected in placebo-treated mice. Amino acid substitutions in the hemagglutinin receptor-binding site at aa 137 or 225 were detected in cloned resistant isolates. A substitution in the neuraminidase (NA) active site (Arg292Lys) was detected in the cloned virus recovered from an oseltamivir-treated mouse. This model would be useful for elucidation of the molecular mechanisms of resistance to NA inhibitors and for testing of anti influenza therapy options that might prevent the emergence of resistant variants. PMID- 16479510 TI - Quantitative analysis of Kaposi sarcoma-associated herpesvirus (KSHV) in KSHV associated diseases. AB - BACKGROUND: Accurate numbers of copies of Kaposi sarcoma-associated herpesvirus (KSHV) and numbers of virus-infected cells in lesions caused by KSHV-associated diseases are unknown. METHODS: Quantitative polymerase chain reaction (PCR) and computerized imaging of immunohistochemical analysis were performed on pathologic sections of samples from persons with KSHV-associated diseases. RESULTS: Real time PCR and semiquantitative PCR-Southern blotting demonstrated that DNA extracted from biopsy samples of KS lesions contained approximately 1-2 viral copies/cell. KSHV-associated lymphoma contained 10-50 viral copies/cell. Computerized-image analysis demonstrated that approximately 49% of cells expressed KSHV-encoded latency-associated nuclear antigen in KS biopsy samples. On the basis of results of real-time PCR and computerized-image analysis, the predicted number of viral copies was 3.2 viral copies/cell in KS lesions. Computerized-image analysis also revealed that the expression of open-reading frame (ORF)-50 protein, an immediate early protein of KSHV, was very rare in KS lesions, which implies that they were mainly composed of proliferating cells latently infected with KSHV. In multicentric Castleman disease lesions, 25% of virus-infected cells expressed ORF50 protein, which suggests the frequent lytic replication of KSHV. CONCLUSIONS: Numbers of viral copies and of virus-positive cells vary among KSHV-associated diseases, which suggests different mechanisms of viral pathogenesis. The combination of real-time PCR and computerized-image analysis provides a useful tool for the assessment of the number of viral copies in KSHV-associated diseases. PMID- 16479511 TI - Preconceptional primary human cytomegalovirus infection and risk of congenital infection. AB - The risk of vertical transmission of human cytomegalovirus (HCMV) was investigated in 14 women who had primary HCMV infection 2-18 weeks before their last menstrual period during 2001-2004. One (8.3%) of 12 newborns examined at birth was found to be subclinically infected. Preconceptional primary HCMV infection has a transmission rate that is significantly higher than that observed in an unselected newborn population but significantly lower than that associated with maternal infection during pregnancy. Although a safe time interval between primary infection and initiation of pregnancy still needs to be defined, documented information can now be provided during counseling. PMID- 16479512 TI - Human cytomegalovirus-encoded alpha -chemokines exhibit high sequence variability in congenitally infected newborns. AB - Most congenital human cytomegalovirus (HCMV) infections are asymptomatic, but some lead to severe disease. We hypothesized that differences in disease manifestations may be partially explained by differences in viral strains. We recently reported an association between unique long (UL) 144 gene polymorphisms and clinical disease. We now report on the sequence heterogeneity of 2 potential HCMV virulence genes that encode alpha -chemokines: UL146 and UL147. These 2 genes were highly divergent in cultured isolates obtained from 23 newborns with congenital HCMV infection and were difficult to categorize. Unlike our findings for the contiguous UL144 gene, no specific UL146 or UL147 genotype was associated with disease outcome. PMID- 16479513 TI - Two-year prospective study of the humoral immune response of patients with severe acute respiratory syndrome. AB - In a cohort study of 56 convalescent patients with severe acute respiratory syndrome (SARS), titers of immunoglobulin G (IgG) antibodies and neutralizing antibodies (NAbs) against SARS-associated coronavirus were assessed at regular intervals (at 1, 4, 7, 10, 16, and 24 months after the onset of disease) by use of enzyme-linked immunosorbent assay and neutralization assay. IgG antibody and NAb titers were highly correlated, peaking at month 4 after the onset of disease and decreasing thereafter. IgG antibodies remained detectable in all patients until month 16, and they became undetectable in 11.8% of patients at month 24. The finding that NAbs remained detectable throughout follow-up is reassuring in terms of protection provided against reinfection; however, NAb titers decreased markedly after month 16. PMID- 16479514 TI - Comparison of an anti-rabies human monoclonal antibody combination with human polyclonal anti-rabies immune globulin. AB - The World Health Organization estimates human mortality from endemic canine rabies to be 55,000 deaths/year. Limited supply hampers the accessibility of appropriate lifesaving treatment, particularly in areas where rabies is endemic. Anti-rabies antibodies are key to protection against lethal rabies. Currently, only human and equine polyclonal anti-rabies immune globulin (HRIG and ERIG) is available. Replacement of HRIG and ERIG with a safer and more widely available product is recommended. We have recently identified a combination of 2 human monoclonal antibodies (MAbs), CR57 and CR4098, that has high potential. We here describe a head-to-head comparison between an CR57/CR4098 MAb cocktail and HRIG. The MAb cocktail neutralized all viruses from a panel of 26 representative street rabies virus isolates. In combination with vaccine, the MAb cocktail protected Syrian hamsters against lethal rabies when administered 24 h after exposure, comparable with the results obtained with HRIG. Furthermore, the MAb cocktail did not interfere with rabies vaccine differently from HRIG. These results demonstrate that the human MAb cocktail of CR57 and CR4098 is a safe and efficacious alternative to RIG in rabies postexposure prophylaxis. PMID- 16479516 TI - Neisseria meningitidis serogroups A and W-135: carriage and immunity in Burkina Faso, 2003. AB - OBJECTIVES: We sought to describe Neisseria meningitidis immunity and its association with pharyngeal carriage in Burkina Faso, where N. meningitidis serogroup W-135 and serogroup A disease are hyperendemic and most of the population received polysaccharide A/C vaccine during 2002. METHODS: We collected oropharyngeal swab samples from healthy residents of Bobo-Dioulasso (4-14 years old, n=238; 15-29 years old, n=250) monthly during February-June 2003; N. meningitidis isolates were analyzed using polymerase chain reaction and serogrouped using immune sera. Serum samples were collected at the first and last clinic visit and analyzed for anti-A, anti-C, anti-W-135, and anti-Y immunoglobulin G (IgG) concentrations and anti-A and anti-W-135 bactericidal titers. RESULTS: N. meningitidis was carried at least once by 18% of participants; this carriage included strains from serogroups W-135 (5%) and Y and X (both <1%) but not from serogroups A, B, or C. At baseline, the prevalence of putatively protective specific IgG concentrations (> or =2 microg/mL) and bactericidal titers (> or =8) was 85% and 54%, respectively, against serogroup A, and 6% and 22%, respectively, against serogroup W-135. Putatively protective anti W-135 IgG concentrations and bactericidal titers were of short duration and were not associated with carriage. CONCLUSION: N. meningitidis serogroup W-135 strains did not induce immunity, despite their circulation. Carriage of serogroup A strains was rare despite the hyperendemic incidence of serogroup A meningitis during 2003 in Bobo-Dioulasso. A vaccine that includes serogroup W-135 antigen and eliminates serogroup A carriage is needed for sub-Saharan Africa. PMID- 16479515 TI - Stability of CXCL-8 and related AU-rich mRNAs in the context of hepatitis C virus replication in vitro. AB - BACKGROUND: Previous studies have shown that levels of CXCL-8 are elevated in patients with chronic hepatitis C virus (HCV) infection and are highest in nonresponders to interferon (IFN) therapy and that CXCL-8 inhibits IFN antiviral action. CXCL-8 expression involves AU-rich elements (AREs) in 3' untranslated regions that regulate mRNA stability. CXCL-8 mRNA stability was, therefore, investigated in the context of HCV replication in 4 replicon cell lines, Huh7 and Huh7.5 cells, and primary human fetal hepatocytes. METHODS: The half-life of CXCL 8 mRNA was measured by use of real-time reverse-transcription polymerase chain reaction following tumor necrosis factor (TNF)- alpha induction in the presence and absence of inhibitors of transcription and translation. Cellular mRNAs containing AREs were assessed by custom microarray analyses. RESULTS: The half life of CXCL-8 mRNA increased in 3 of 4 HCV replicon cell lines, particularly after treatment with TNF- alpha , a potent inducer of CXCL-8. CXCL-8 mRNA was superinduced by TNF- alpha in the presence of the protein-synthesis inhibitor cycloheximide. Analysis of >1500 ARE-containing cellular mRNAs, by use of microarrays, revealed that CXCL-8 and other newly identified ARE genes were induced by TNF- alpha in Huh7 cells and were coordinately regulated. CONCLUSION: The data suggest that increased CXCL-8 gene expression in the context of HCV replication involves posttranscriptional events. PMID- 16479517 TI - Antibody persistence 3 years after immunization of adolescents with quadrivalent meningococcal conjugate vaccine. AB - BACKGROUND: A quadrivalent meningococcal conjugate vaccine (MCV-4) is recommended for United States teenagers. The duration of protective immunity is unknown. We investigated serum antibody persistence 3 years after vaccination of adolescents. METHODS: Serum samples from participants of a randomized trial who had received MCV-4 (n=52) or polysaccharide vaccine (MPSV-4; n=48) and from unvaccinated controls (n=60) were assayed for serogroups C, W-135, and Y anticapsular antibody concentrations by use of a radioantigen binding assay and for bactericidal activity (in a human complement assay) and passive protection against serogroup C bacteremia in an infant rat model. RESULTS: A higher proportion of participants in the vaccine groups had protective bactericidal titers (> or =1 : 4), compared with that in the control group (for MCV-4 recipients vs. controls, P<.01; for MPSV-4 recipients vs. controls, P< or =.06). More MCV-4 recipients had W-135 bactericidal titers > or =1 : 4 than did MPSV-4 recipients (P=.01). More MCV-4 recipients had passive protective activity against serogroup C bacteremia than did MPSV-4 recipients (76% vs. 49%; P<.01). The differences in protective activity were largest between participants in the vaccine groups with bactericidal titers <1 : 4 (63% protective in MCV-4 recipients vs. 31% protective in MPSV-4 recipients; P=.01). CONCLUSIONS: Compared with MPSV-4, MCV-4 elicited greater persistence of antibody activity against serogroups C and W-135 at 3 years after vaccination in adolescents. On the basis of passive protection data in an infant rat model, bactericidal titers > or =1 : 4 underestimate protective immunity. PMID- 16479519 TI - Characterization of the Staphylococcus epidermidis accessory-gene regulator response: quorum-sensing regulation of resistance to human innate host defense. AB - Staphylococci are important opportunistic pathogens. However, there is a lack of information on how these bacteria survive inside the human body during infection. This study demonstrates that quorum-sensing regulation in Staphylococcus epidermidis protects it from key mechanisms of human innate host defense. To gain a better understanding of the basis of the observed phenotype, the agr quorum sensing regulon of S. epidermidis was characterized by a genomewide analysis of gene expression. The gene-expression data indicate that agr adapts bacterial physiology to stationary growth and, furthermore, that it controls a series of virulence factors, including degradative exoenzymes possibly involved in resistance to antimicrobial peptides. Remarkably, agr also regulates general and oxidative stress-response factors, including detoxifying enzymes of reactive oxygen species. Taken together, the results of this study indicate that quorum sensing regulation in staphylococci has important, previously unknown functions that contribute to protection from mechanisms of human innate host defense--and, therefore, to the pathogen's survival in the human host. PMID- 16479518 TI - Sublethal doses of Bacillus anthracis lethal toxin inhibit inflammation with lipopolysaccharide and Escherichia coli challenge but have opposite effects on survival. AB - BACKGROUND: On the basis of the findings of previous in vitro studies, we hypothesized that anthrax lethal toxin (LeTx) would have anti-inflammatory effects in vivo. METHODS: We investigated the effects of sublethal doses of LeTx in rats receiving intravascular challenge with lipopolysaccharide (LPS) or intratracheal challenge with Escherichia coli. RESULTS: In rats receiving 24-h infusions of LPS, compared with control rats, pretreatment with high or low sublethal doses of LeTx 3 h before infusion produced similar patterns of reduction in the hazards ratio (HR) of survival at 168 h (0.60 [95% confidence interval {CI}, 0.37-0.98]; P=.03, for the doses combined). LeTx increased mean arterial blood pressure throughout the period of LPS infusion (P=.001); decreased the levels of 10 of 13 cytokines assessed (i.e., interleukin [IL]-1 alpha , IL-1 beta , IL-2, IL-4, IL-6, IL-10, interferon- gamma , tumor necrosis factor- alpha , granulocyte macrophage-colony-stimulating factor, migratory inhibitory protein [MIP]-1 alpha , MIP-2, MIP-3 alpha , and RANTES) at 2 h; decreased all 13 cytokine levels at 8 h; decreased only 4 cytokine levels at 24 h; and decreased the plasma level of nitric oxide (NO) at 8 h and 24 h but not at 2 h (P< or =.02, for the effect of LeTx, across time, on both cytokine levels and the NO level). Although pretreatment with LeTx before challenge with E. coli altered mean arterial blood pressure, cytokine levels, and the NO level in a pattern similar to that noted in association with LPS infusion, it increased the HR in a pattern different from that associated with LPS (4.36 [95% CI, 0.3-63.4]; P=.04, for the effects of LeTx with LPS vs. E. coli). CONCLUSION: Inhibition of inflammation with LeTx can occur in vivo, and, although beneficial with noninfectious stimuli, it may be harmful with bacteria. PMID- 16479520 TI - Borrelia burgdorferi lipoprotein-mediated TLR2 stimulation causes the down regulation of TLR5 in human monocytes. AB - Toll-like receptors (TLRs) trigger innate immune responses via the recognition of conserved pathogen-associated molecular patterns. Lipoproteins from Borrelia burgdorferi, the agent of Lyme disease, activate inflammatory cells through TLR2 and TLR1. We show that stimulation of human monocytes with B. burgdorferi lysate, lipidated outer surface protein A, and triacylated lipopeptide Pam3CysSerLys4 results in the up-regulation of both TLR2 and TLR1 but the down-regulation of TLR5, the receptor for bacterial flagellin, and that this effect is mediated via TLR2. TLR4 stimulation had no effect on TLR2, TLR1, and TLR5 expression. Human monocytes stimulated with TLR5 ligands (including p37 or flaA, the minor protein from B. burgdorferi flagella) up-regulated TLR5. In addition, TLR2 stimulation rendered cells hyporesponsive to a TLR5 agonist. These results indicate that diverse stimuli can cause differential TLR expression, and we hypothesize that these changes may be useful for either the pathogen and/or the host. PMID- 16479521 TI - Effects of a single large dose of vitamin A, given during the postpartum period to HIV-positive women and their infants, on child HIV infection, HIV-free survival, and mortality. AB - BACKGROUND: Low maternal serum retinol level is a risk factor for mother-to-child transmission (MTCT) of human immunodeficiency virus (HIV). Multiple-large-dose vitamin A supplementation of HIV-positive children reduces mortality. The World Health Organization recommends single-large-dose vitamin A supplementation for postpartum women in areas of prevalent vitamin A deficiency; neonatal dosing is under consideration. We investigated the effect that single-large-dose maternal/neonatal vitamin A supplementation has on MTCT, HIV-free survival, and mortality in HIV-exposed infants. METHODS: A total of 14,110 mother-infant pairs were enrolled < or =96 h after delivery, and both mother and infant, mother only, infant only, or neither received vitamin A supplementation in a randomized, placebo-controlled trial with a 2 x 2 factorial design. All but 4 mothers initiated breast-feeding. A total of 4495 infants born to HIV-positive women were included in the present analysis. RESULTS: Neither maternal nor neonatal vitamin A supplementation significantly affected postnatal MTCT or overall mortality between baseline and 24 months. However, the timing of infant HIV infection modified the effect that supplementation had on mortality. Vitamin A supplementation had no effect in infants who were polymerase chain reaction (PCR) positive [corrected] for HIV at baseline. In infants who were PCR negative at baseline and PCR positive at 6 weeks, neonatal supplementation reduced mortality by 28% (P=.01), but maternal supplementation had no effect. In infants who were PCR negative at 6 weeks, all 3 vitamin A regimens were associated with ~2-fold higher mortality (P< or =.05). CONCLUSIONS: Targeted vitamin A supplementation of HIV-positive children prolongs their survival. However, postpartum maternal and neonatal vitamin A supplementation may hasten progression to death in breast-fed children who are PCR negative at 6 weeks. These findings raise concern about universal maternal or neonatal vitamin A supplementation in HIV-endemic areas. PMID- 16479522 TI - Impact of HIV-associated immunosuppression on malaria infection and disease in Malawi. AB - BACKGROUND: Human immunodeficiency virus (HIV) infection and malaria coexist in much of Africa. Previous studies differ in their findings on the interactions between the 2 infections. METHODS: Adults living with HIV infection in Blantyre, Malawi, were enrolled in a longitudinal observational study from September 2002 to August 2004. Malaria blood smears were obtained monthly and for any illness suggestive of malaria. Complete evaluations of all illness episodes were conducted, regardless of malaria smear results. RESULTS: The incidence of clinical malaria episodes was higher in participants with CD4 cell counts <200 cells/mm3 than in those with CD4 cell counts >500 cells/mm3. The trend was preserved when increasingly specific definitions of malaria disease were used. The prevalence of malaria infection was not associated with CD4 cell count. In per-visit analysis, lower CD4 cell counts were associated with higher incidences of pneumonia, sepsis, and tuberculosis but not of malaria. Severe malaria was rare, with only 3 cases in 591 person-years of observation. Parasite density and CD4 cell count were independent risk factors for fever. CONCLUSIONS: Profoundly immunosuppressed adults with HIV infection require more-frequent treatment for uncomplicated malaria, but malaria infection and disease are less strongly associated with HIV-associated immunosuppression than are other opportunistic infections. Where malaria is common, the high incidence of fever found among immunosuppressed adults may lead to misclassification of illness episodes as malaria. PMID- 16479523 TI - CD8+ T cells in HIV disease exhibit cytokine receptor perturbation and poor T cell receptor activation but are responsive to gamma-chain cytokine-driven proliferation. AB - BACKGROUND: Cytokines are important for inducing T cell maturation, proliferation, and survival. Despite the known dysregulation of cytokines in human immunodeficiency virus (HIV) infection, cytokine receptor expression is relatively unexplored. METHODS: We examined maturation markers (naive, central memory, effector memory, and effector); the cytokine receptors interleukin (IL) 2R beta , common gamma (C gamma ) chain, IL-7R alpha , IL-15R alpha; and proliferative responses of T cells in a cohort of HIV-infected pediatric patients (median age, 14.82 years) receiving antiretroviral therapy, arbitrarily designated as immunologic responders (group I) and nonresponders (group II) on the basis of a CD4+ T cell count cutoff of 25%. RESULTS: Patients had increased percentages of effector memory CD8+ T cells, in comparison with those in healthy control subjects, with reduced expression of IL-7R alpha in the central memory and effector memory subsets and of the C gamma chain in all maturation subsets of CD8+ T cells. IL-7R alpha +CD8+ T cell percentages were directly correlated with CD4+ T cell percentages. In immunologic nonresponders, anti-CD3+ or HIV Gag antigen-induced CD8+ T cell proliferation was impaired, but proliferation in response to the homeostatic cytokines IL-2 and IL-15 was preserved.Conclusions. Cytokine receptor deficiencies may contribute to immune deficiency in HIV infected patients, and gamma -chain-utilizing cytokines may play an important role in vivo in maintaining the memory subsets of T cells in patients with CD4+ T cell deficiency. PMID- 16479526 TI - Age dependence of the relation between reassortant rotavirus vaccine (RotaShield) and intussusception. PMID- 16479524 TI - Pulmonary V beta 4+ T cells from Histoplasma capsulatum-infected mice respond to a homologue of Sec31 that confers a protective response. AB - The population of V beta 4+ T cells expands in the lungs of C57BL/6 mice infected with Histoplasma capsulatum, and the elimination of these cells impairs protective immunity. To determine the antigen or antigens that trigger their proliferation, V beta 4+ T cell hybridomas were generated from the lungs and spleens of infected mice. We mapped the antigenic determinants by T cell Western blot. Pulmonary and splenic T cells recognized 3 regions comprising <25, 55-70, and 125-140 kDa. The majority of hybridomas from lungs, but not from spleens, responded to the high molecular mass region. A protein from that area was identified, by amino acid sequencing, as a homologue of Sec31 from Saccharomyces cerevisiae. Vaccination with recombinant Sec31 reduced fungal burden and improved survival in mice, and its efficacy was critically dependent on the presence of V beta 4+ T cells. Thus, a homologue of Sec31 is a trigger of the expansion of the V beta 4+ T cell population and is important to the generation of protective immunity. PMID- 16479527 TI - Human illness and isolation of low-pathogenicity avian influenza virus of the H7N3 subtype in British Columbia, Canada. PMID- 16479529 TI - Coinfection with Borrelia burgdorferi and Babesia microti: bad or worse? PMID- 16479531 TI - Poverty and the spread of bloodborne disease in central China. PMID- 16479532 TI - Quantitative glutamate spectroscopic imaging of the human hippocampus. AB - We have evaluated a three-dimensional localized spectroscopic imaging sequence that uses two pairs of adiabatic full-passage pulses, which optimizes the detection of glutamate resonances at moderate echo times. This sequence provides excellent volume localization while simultaneously reducing J-modulation losses of glutamate. We have simulated the performance of this sequence for glutamate and used it to quantitatively measure glutamate in the human hippocampus using a linear components model. Using tissue segmentation and regression analysis, we measured a glutamate concentration of 8.8 +/- 2.1 mM in hippocampal and temporal gray matter and 3.7 +/- 1.1 mM in temporal white matter (95% CI). We have used this approach in a small group of patients (n = 5) with unilateral hippocampal epilepsy. PMID- 16479533 TI - Attenuation of mouse acute colitis by naked hepatocyte growth factor gene transfer into the liver. AB - BACKGROUND: Hepatocyte growth factor (HGF) has multiple biological effects on a wide variety of cells. It modulates intestinal epithelial proliferation and migration, and critically regulates intestinal wound healing. AIMS: To investigate the therapeutic effect of HGF gene transfer, we introduced the HGF gene into the liver of mice with acute colitis. METHODS: The rat HGF expression plasmid vector, pCAGGS-HGF, was injected via the tail vein into C57BL/6 mice, followed by dosing with dextran sulfate sodium in distilled water. Firstly, the HGF gene was injected once on day 0. Secondly, the HGF gene was injected on day 0 and again on day 2. RESULTS: Injection of the HGF gene ameliorated colitis with inhibition of both loss of body weight and shortening of colon length. It protected the colon from epithelial erosions and cellular infiltration. Expression of mRNAs for IFN-gamma, IL18, and TNF-alpha was reduced in the colon. In contrast, expression of mRNA for IL-10 was increased. The numbers of BrdU positive intestinal epithelial cells were increased, and the numbers of TUNEL positive apoptotic cells were decreased. Furthermore, a second injection prolonged the elevation of serum HGF levels, and ameliorated the symptoms better than a single injection. The empty pCAGGS plasmid did not ameliorate acute colitis. CONCLUSIONS: HGF gene transfer attenuated acute colitis by facilitating intestinal wound repair as well as inhibiting inflammation, suggesting a new strategy for treatment of IBD. PMID- 16479534 TI - A broad-spectrum fluorescence-based peptide library for the rapid identification of protease substrates. AB - Identification of peptide substrates for proteases can be a major undertaking. To overcome issues such as feasibility and deconvolution, associated with large peptide libraries, a 'small but smart' generic fluorescence resonance energy transfer rapid endopeptidase profiling library (REPLi) was synthesised as a tool for rapidly identifying protease substrates. Within a tripeptide core, flanked by Gly residues, similar amino acids were paired giving rise to a relatively small library of 3375 peptides divided into 512 distinct pools each containing only 8 peptides. The REPLi was validated with trypsin, pepsin, the matrix metalloprotease (MMP)-12 and MMP-13 and calpains-1 and -2. In the case of calpain 2, a single iteration step involving LC-MS, provided the definitive residue specificity from which a highly sensitive fluorogenic substrate, (FAM)-Gly-Gly Gly-Gln-Leu-Tyr-Gly-Gly-DPA-Arg-Arg-Lys-(TAMRA), was then designed. The thorough validation of this 'small but smart' peptide library with representatives from each of the four mechanistic protease classes indicates that the REPLi will be useful for the rapid identification of substrates for multiple proteases. PMID- 16479535 TI - Analysis of the wheat and Puccinia triticina (leaf rust) proteomes during a susceptible host-pathogen interaction. AB - Wheat leaf rust is caused by the fungus Puccinia triticina. The genetics of resistance follows the gene-for-gene hypothesis, and thus the presence or absence of a single host resistance gene renders a plant resistant or susceptible to a leaf rust race bearing the corresponding avirulence gene. To investigate some of the changes in the proteomes of both host and pathogen during disease development, a susceptible line of wheat infected with a virulent race of leaf rust were compared to mock-inoculated wheat using 2-DE (with IEF pH 4-8) and MS. Up-regulated protein spots were excised and analyzed by MALDI-QqTOF MS/MS, followed by cross-species protein identification. Where possible MS/MS spectra were matched to homologous proteins in the NCBI database or to fungal ESTs encoding putative proteins. Searching was done using the MASCOT search engine. Remaining unmatched spectra were then sequenced de novo and queried against the NCBInr database using the BLAST and MS BLAST tools. A total of 32 consistently up regulated proteins were examined from the gels representing the 9-day post infection proteome in susceptible plants. Of these 7 are host proteins, 22 are fungal proteins of known or hypothetical function and 3 are unknown proteins of putative fungal origin. PMID- 16479536 TI - Relative and absolute quantitative expression profiling of cytochromes P450 using isotope-coded affinity tags. AB - The development of a novel method for absolute quantification of proteins based on isotope-coded affinity tagging using ICAT reagents is described. The method exploits synthetic peptide standards to determine protein content at the femtomole level in biological samples. The approach is generally applicable to any subset of proteins, but is particularly appropriate for quantitative analysis of multiple, closely related isoforms, and for hydrophobic proteins that are poorly represented in 2-D gels. Relative and absolute quantification techniques are applied to an important group of microsomal metabolic enzymes, the cytochromes P450 (P450), which are critical in determining the disposition, safety and efficacy of drugs in man. Measurement of the P450 induction profile in response to chemicals is a fundamental aspect of drug safety evaluation and is currently achieved by low-throughput methods employing poorly discriminatory antibodies or substrates. Tagging technology is shown to supersede conventional methods for P450 profiling in terms of discriminatory power and throughput, exemplified by the simultaneous detection of distinct induction profiles for cyp2c subfamily members in response to phenobarbitone: cyp2c29 expression, but not cyp2c40 or cyp2c50, was induced threefold by treatment. This technology should abbreviate the drug development pathway, and provide a widely applicable, rapid means of quantifying proteins. PMID- 16479537 TI - Systematic localisation of proteins fused to the green fluorescent protein in Bacillus subtilis: identification of new proteins at the DNA replication factory. AB - Construction and microscopic imaging of protein fusions to green fluorescent protein (GFP) have revolutionised our understanding of bacterial structure and function. We have undertaken a systematic study of the localisation of over 100 Bacillus subtilis proteins, following the development of high-throughput construction and analysis procedures. We focused on proteins linked in various ways to the DNA replication machinery, as well as on proteins exemplifying a range of other cellular functions and structures. The results validate the approach as a way of obtaining systematic protein localisation information. They also provide a range of novel biological insights, particularly through the identification of a number of proteins not previously known to be associated with the DNA replication factory. PMID- 16479538 TI - Spontaneous mutations in the human CMV HLA class I homologue UL18 affect its binding to the inhibitory receptor LIR-1/ILT2/CD85j. AB - Human cytomegalovirus (HCMV) down-regulates cell surface expression of HLA class I molecules (HLA-I). UL18, an HCMV-encoded HLA-I homologue, has been proposed to protect virus-infected cells against NK cell recognition by engaging the inhibitory receptor leukocyte Ig-like receptor (LIR)-1, which also binds a broad spectrum of HLA-I alleles, including HLA-G1. Because genetic and biological differences exist among HCMV strains, we characterized laboratory (AD169) and clinical (4636, 13B, 109B) strain-derived UL18 proteins. Compared to the known AD169-derived UL18, mutations were found in clinical strain-derived UL18. They were clustered in the alpha3 domain (13B), previously shown to be critical for LIR-1 binding, or in the alpha1 domain (4636). Iotan cytotoxicity assays, pretreatment of LIR-1+ NKL with soluble 4636-UL18 completely abolished LIR-1 dependent protection from NK lysis, conferred by the expression of HLA-G1 on target cells (721.221-HLA-G1+). Similarly, flow cytometry, Biacore and ELISA experiments showed 4636-UL18 and 13B-UL18 to have the strongest binding capacity to LIR-1. Our results suggest the importance of two independent UL18 regions for LIR-1 binding, one localized on the tip of the alpha3 domain, and another composed of two loops that emerge from the alpha1 domain. Strain variations in these domains may result in different UL18-mediated effects on LIR-1+ cells during the course of HCMV infection. PMID- 16479539 TI - Identification of naturally processed peptides bound to the class I MHC molecule H-2Kd of normal and TAP-deficient cells. AB - This report details the biochemical features of natural peptides selected by the H-2Kd class I MHC molecule. In normal cell lines, the length of the naturally processed peptides ranged from 8 to 18 amino acids, although the majority were 9 mers (16% were longer than nine residues). The binding motif for the 9-mer peptides was dominated by the presence of a tyrosine at P2 and an isoleucine/leucine at the P9 position. The P2 residue contributed most towards binding; and the short peptides bound better and formed longer-lived cell surface complexes than the long peptides, which bound poorly and dissociated rapidly. The longer peptides did not exhibit this strictly defined motif. Trimming the long peptides to their shorter forms did not enhance binding and conversely, extending the 9-mer peptides did not decrease binding. The long peptides were present on the cell-surface bound to H-2Kd (Kd) and were not intermediate products of the class I MHC processing pathway. Finally, in two different TAP-deficient cells the long peptides were the dominant species, which suggested that TAP-independent pathways selected for long peptides by class I MHC molecules. PMID- 16479540 TI - Alpha-melanocyte-stimulating hormone down-regulates CXC receptors through activation of neutrophil elastase. AB - Considering the role of interleukin-8 (IL-8) in a large number of acute and chronic inflammatory diseases, the regulation of IL-8-mediated biological responses is important. Alpha-melanocyte-stimulating hormone (alpha-MSH), a tridecapeptide, inhibits most forms of inflammation by an unknown mechanism. In the present study, we have found that alpha-MSH interacts predominantly with melanocortin-1 receptors and inhibits several IL-8-induced biological responses in macrophages and neutrophils. It down-regulated receptors for IL-8 but not for TNF, IL-4, IL-13 or TNF-related apoptosis-inducing ligand (TRAIL) in neutrophils. It down-regulated CXCR type 1 and 2 but not mRNA levels. alpha-MSH did not inhibit IL-8 binding in purified cell membrane or affinity-purified CXCR. IL-8 or anti-CXCR Ab protected against alpha-MSH-mediated inhibition of IL-8 binding. The level of neutrophil elastase, a specific serine protease, but not cathepsin G or proteinase 3 increased in alpha-MSH-treated cells, and restoration of CXCR by specific neutrophil elastase or serine protease inhibitors indicates the involvement of elastase in alpha-MSH-induced down-regulation of CXCR. These studies suggest that alpha-MSH inhibits IL-8-mediated biological responses by down-regulating CXCR through induction of serine protease and that alpha-MSH acts as a potent immunomodulator in neutrophil-driven inflammatory distress. PMID- 16479541 TI - Proteasome inhibitor-induced apoptosis in human monocyte-derived dendritic cells. AB - Proteasome inhibitors possess potent antitumor activity against a broad spectrum of human malignancies. However, the effects of these compounds on the immune system still have to be clearly determined. In the present study, we have investigated the effects of proteasome inhibitors on dendritic cells (DC), antigen-presenting cells playing a key role in the initiation of immune responses. Exposure to the proteasome inhibitors bortezomib, MG132 or epoxomicin was found to promote apoptosis of human monocyte-derived DC and to reduce the yield of viable DC when given to monocytes early during differentiation to DC. DC apoptosis via proteasome inhibition was accompanied by mitochondria disruption and subsequent activation of the caspase cascade. Up-regulation and intracellular redistribution of Bcl-2-associated X protein (Bax), a pro-apoptotic Bcl-2 family protein, were observed in DC treated with these compounds and represent a suitable mechanism leading to activation of the intrinsic apoptotic pathway. Finally, active protein synthesis was found to represent an upstream prerequisite for DC apoptosis induced by proteasome inhibitors, since the translation inhibitor cycloheximide blocked all of the steps of the observed apoptotic response. In conclusion, induction of apoptosis in DC may represent a novel mechanism by which proteasome inhibitors affect the immune response at the antigen-presenting cell level. PMID- 16479542 TI - Pertussis toxin as an adjuvant suppresses the number and function of CD4+CD25+ T regulatory cells. AB - We observed a remarkable reduction in the frequency and immunosuppressive activity of splenic CD4+CD25+ T cells in C57BL/6 mice with MOG33-55-induced experimental autoimmune encephalomyelitis (EAE). Our study revealed that pertussis toxin (PTx), one component of the immunogen used to induce murine EAE, was responsible for down-regulating splenic CD4+CD25+ cells. Treatment of normal BALB/c mice with PTx in vivo reduced the frequency, suppressive activity and FoxP3 expression by splenic CD4+CD25+ T cells. However, PTx treatment did not alter the expression of characteristic phenotypic markers (CD45RB, CD103, GITR and CTLA-4) and did not increase the expression of CD44 and CD69 by the residual splenic and lymph node CD4+CD25+ T cells. This property of PTx was attributable to its ADP-ribosyltransferase activity. PTx did not inhibit suppressive activity of purified CD4+CD25+ T regulatory (Treg) cells in vitro, but did so in vivo, presumably due to an indirect effect. Although the exact molecular target of PTx that reduces Treg activity remains to be defined, our data suggests that alteration of both distribution and function of splenic immunocytes should play a role. This study concludes that an underlying cause for the immunological adjuvanticity of PTx is down-regulation of Treg cell number and function. PMID- 16479543 TI - Differential mechanisms of microparticle transfer toB cells and monocytes: anti inflammatory propertiesof microparticles. AB - Microparticles are small vesicles released from the plasma membrane of various cell types independently of apoptosis or cell death, are transferred between cells, and carry membrane proteins from one cell to another. We have studied the mechanism of uptake of microparticles by monocytes and B cells. The transfer of microparticles to B cells was almost completely dependent on complement. Incubation of microparticles with serum resulted in opsonization of microparticles with the complement cleavage product iC3b. The subsequent transfer to B cells was mediated by the complement receptor CR2. The interaction between iC3b-opsonized microparticles and B cells reduced the activation of B cells as measured by expression of MHC class II, CD86 and CD25. In contrast, transfer of microparticles to monocytes was only partially complement dependent, but involved calcium and annexin V, and was found to change the cytokine profile of monocytes towards a reduced release of the pro-inflammatory cytokines GM-CSF and TNF-alpha and an increased release of the anti-inflammatory cytokine IL-10. These data show that microparticles are taken up by B cells and monocytes by different mechanisms and modulate the activation of monocytes and B cells towards an anti-inflammatory phenotype. Microparticles might be involved in counterbalancing pro-inflammatory signals arising from tissue injury or inflammation. PMID- 16479544 TI - Epstein-Barr virus (EBV) latent membrane protein-1-specific cytotoxic T lymphocytes targeting EBV-carrying natural killer cell malignancies. AB - Epstein-Barr virus (EBV)-encoded latent membrane protein (LMP) 1 is a potential target for immunotherapy of some proportion of Hodgkin's disease cases, nasopharyngeal carcinomas, EBV-associated natural killer (NK)/T lymphomas, and chronic active EBV infection (CAEBV). Since it is unknown whether EBV-infected NK/T cells are susceptible to lysis by LMP1-specific cytotoxic T lymphohcytes (CTL), we here tested the ability of mRNA-transduced antigen-presenting cells (APC) to stimulate rare LMP1-specific CTL. A 43-amino acid N-terminal deletion mutant LMP1 (DeltaLMP1) could be efficiently expressed in dendritic cells and CD40-activated B cells upon mRNA electroporation. DeltaLMP1-expressing APC were found to stimulate LMP1-specific CTL from a healthy donor and a CTL clone recognized a peptide, IIIILIIFI, presented by HLA-A*0206 molecules. Processing and presentation of the antigenic peptide proved dependent on expression of an immunoproteasome subunit, low-molecular-weight protein-7, as confirmed by RNA interference gene silencing. Furthermore, an EBV-infected NK cell line derived from a patient with CAEBV, and another from an NK lymphoma with enforced HLA A*0206 expression, were specifically lysed by the CTL. Overall, these data suggest that immunotherapy targeting LMP1 in EBV-associated NK lymphomas and CAEBV might serve as an alternative treatment modality. PMID- 16479545 TI - Alternative activation deprives macrophages of a coordinated defense program to Mycobacterium tuberculosis. AB - A potent Th1 immune response is critical to the control of tuberculosis. The impact of an additive Th2 response on the course of disease has so far been insufficiently characterized, despite increased morbidity after co-infection with Mycobacterium tuberculosis and Th2-eliciting helminths and possible involvement of Th2 polarization in reactivation of latent tuberculosis. Here, we describe the gene expression profile of murine bone marrow-derived macrophages alternatively activated by IL-4 in response to infection with M. tuberculosis. Comparison of transcriptional profiles of infected IL-4- and IFN-gamma-activated macrophages revealed delayed and partially diminished responses to intracellular bacteria in alternatively activated macrophages, characterized by reduced exposure to nitrosative stress and increased iron availability, respectively. Alternative activation of host macrophages correlated with elevated expression of the M. tuberculosis iron storage protein bacterioferritin as well as reduced expression of the mycobactin synthesis genes mbtI and mbtJ. The extracellular matrix remodeling enzyme matrix metalloproteinase (MMP)-12 was induced in alternatively activated macrophages in vitro, and MMP-12-expressing macrophages were abundant at late, but not early, stages of tuberculosis in murine lungs. Our findings emphasize that alternative activation deprives macrophages of control mechanisms that limit mycobacterial growth in vivo, thus supporting intracellular persistence of M. tuberculosis. PMID- 16479546 TI - Differential roles for CXCR3 in CD4+ and CD8+ T cell trafficking following viral infection of the CNS. AB - Lymphocyte infiltration into the central nervous system (CNS) following viral infection represents an important component of host defense and is required for control of viral replication. However, the mechanisms governing inflammation in response to viral infection of the CNS are not well understood. Following intracranial (i.c.) infection of susceptible mice with mouse hepatitis virus (MHV), mice develop an acute encephalomyelitis followed by a chronic demyelinating disease. The CXC chemokine ligand 10 (CXCL10) is expressed following MHV infection and signals T cells to migrate into the CNS. The functional contribution of the CXCL10 receptor CXCR3 in host defense and disease in response to MHV infection was evaluated. The majority of CD4+ and CD8+ T cells infiltrating the CNS following MHV infection express CXCR3. Administration of anti-CXCR3 antibody reduced CD4+ T cell infiltration (p